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Detection of transformed cells in crown gall tumors using the GUS reporter gene and correlation of GUS stained cells with T-DNA gene activity  

SciTech Connect

Crown gall tumors are a mixture of transformed hormone producing cells and normal cells. Until now it has not been possible to directly visualize these cell types in situ. We have constructed strains of Agrobacterium tumefaciens that carry the 35S-{beta}-glucuronidase (GUS) reporter gene in either wild type or mutant Ti plasmids. Using histochemical staining for GUS activity, blue (GUS positive) sectors are observed in tumor sections. In order to demonstrate that the blue sectors actually represent cells expressing other T-DNA genes, we have looked for T-DNA gene encoded enzyme activity in the stained and unstained sectors. The blue sectors accumulate octopine (a product of the octopine synthase gene on the T-DNA) while the white (GUS negative) sectors do not. We conclude that the use of the GUS reporter gene provides a sensitive and reliable method for visualizing transformation events in plant tissues. A comparison of the proportion of transformed and nontransformed cells in wild type tumors vs. tumors deficient in auxin or cytokinin encoding genes will be discussed.

Black, R.C. (Pennsylvania State Univ., Media (USA)); Labriola, J.; Binns, A.N. (Univ. of Pennsylvania, Philadelphia (USA))



Transient reporter gene (GUS) expression in creeping bentgrass ( Agrostis palustris ) is affected by in vivo nucleolytic activity  

Microsoft Academic Search

Leaf and callus tissues of a creeping bentgrass cultivar (Penn A4) had high nuclease activities that degraded exogenously added plasmid DNA. When callus tissue was incubated for 24 h with heparin, spermidine, aurintricarboxylic acid or polyethylene glycol, only heparin and spermidine were effective as in vitro nuclease inhibitors, protecting exogenously added plasmid DNA from degradation. When ß-glucuronidase (GUS) reporter gene activity

Chhandak Basu; Hong Luo; Albert P. Kausch; Joel M. Chandlee



Transient reporter gene (GUS) expression in creeping bentgrass (Agrostis palustris) is affected by in vivo nucleolytic activity.  


Leaf and callus tissues of a creeping bentgrass cultivar (Penn A4) had high nuclease activities that degraded exogenously added plasmid DNA. When callus tissue was incubated for 24 h with heparin, spermidine, aurintricarboxylic acid or polyethylene glycol, only heparin and spermidine were effective as in vitro nuclease inhibitors, protecting exogenously added plasmid DNA from degradation. When beta-glucuronidase (GUS) reporter gene activity was evaluated in heparin-treated (0.6%), 14-month old callus following microprojectile bombardment, GUS activity increased 1000-fold compared to equivalent aged untreated Penn A4 callus. Similar enhancement from heparin pretreatment (0.6% or 1.2%) was not observed in 6-month old callus. This is likely due to much higher activities of nuclease in the younger callus. PMID:12889827

Basu, Chhandak; Luo, Hong; Kausch, Albert P; Chandlee, Joel M



Transient expression of exogenous gus gene in Porphyra yezoensis (Rhodophyta)  

NASA Astrophysics Data System (ADS)

Electroporation, PEC, PEG plus electroporation and Biolistics methods were tested in gene transformation of P. yezoensis. The exogenous gus was from plasmid of pBI121 and pCAMBIA1301, both contain the CaMV35S promoter. The receptors included the protoplasts, tissues and free-living conchocelis filaments of P. yezoensis. Several factors, for example, the voltage, capacitance and bivalent cations, etc., were studied. Results show that these four methods are all efficient for gene transformation in P. yezoensis; and that PEG is the best one, with transformation efficiency of up to 4×10-5. GUS activity was detected 26 days after transformation by using PEG method.

Kuang, Mei; Wang, Su-Juan; Li, Yao; Shen, Da-Leng; Zeng, Cheng-Kui



Construction of improved vectors and cassettes containing gusA and antibiotic resistance genes for studies of transcriptional activity and bacterial localization  

Microsoft Academic Search

Broad-host-range, conjugative vectors with a constitutively expressed gusA gene combined with different antibiotic resistance (tetracycline, gentamicin, kanamycin) genes have been constructed. These plasmids are designed for tracking Gram-negative bacterial strains without the risk of random mutagenesis. We also constructed a set of cassettes containing a promoterless gusA gene linked with different antibiotic resistance genes for making transcriptional fusions and for

Jerzy Wielbo; Anna Skorupska



Promoterless gus gene shows leaky ?-glucuronidase activity during transformation of tomato with bspA gene for drought tolerance  

Microsoft Academic Search

Transformation of tomato (Lycopersicon esculentum Mill.) was carried out using disarmed Agrobacterium tumefaciens strain EHA 105 harboring a binary vector pBIG-HYG-bspA. The plasmid contains the bspA (boiling stable protein of aspen) gene under the control of a CaMV35S promoter and nopaline synthase (NOS) terminator, hygromycin phosphotransferase gene (hpt) driven by nopaline synthase promoter and polyadenylation signal of Agrobacterium gene7 as

R. Roy; R. S. Purty; V. Agrawal; S. C. Gupta



Using the pER8:GUS reporter system to screen for phytoestrogens from Caesalpinia sappan.  


Arabidopsis thaliana pER8:GUS, a low-cost, highly efficient, and convenient transgenic plant system, was used to assay the estrogen-like activity of 30 traditional Chinese medicines. The MeOH extract of Caesalpinia sappan exhibited significant bioactivity in this assay, and subsequent bioactivity-guided fractionation of the extract led to the isolation of one new compound, (S)-3,7-dihydroxychroman-4-one (1), and 10 known compounds. Both the plant pER8:GUS and in vitro estrogen response element reporter assays were used to evaluate the estrogenic activity of the isolated compounds, and these two systems produced comparable results. Compounds 6, 8, and 11 showed significant estrogenic activity comparable to genistein. These active compounds were determined to be nontoxic new sources of phytoestrogens. In addition, compounds 2 and 3 inhibited ERE transcription induced by 17?-estradiol. A docking model revealed that compounds 6, 8, and 11 showed high affinity to the estrogen receptor. The pER8:GUS reporter system was demonstrated to be a useful and effective technique in phytoestrogen discovery. PMID:21800859

Lai, Wan-Chun; Wang, Hui-Chun; Chen, Guan-Yu; Yang, Juan-Cheng; Korinek, Michal; Hsieh, Chia-Jung; Nozaki, Hiroshi; Hayashi, Ken-Ichiro; Wu, Chih-Chung; Wu, Yang-Chang; Chang, Fang-Rong



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



NFL Veterans Show 'Abnormalities' in Brain Activity  


... please enable JavaScript. NFL Veterans Show 'Abnormalities' in Brain Activity: Study Ex-players who had been sent ... researchers say they see unusual activity in the brains of former National Football League players. These traits ...


Detailed characterization of the posttranscriptional gene-silencing-related small RNA in a GUS gene-silenced tobacco.  

PubMed Central

Posttranscriptional gene-silencing phenomena such as cosuppression and RNA interference are associated with the occurrence of small, about 21-23 nt short RNA species homologous to the silenced gene. We here show that the small RNA present in silenced transgenic plants can easily be detected in total RNA isolated according to standard procedures. This will allow for the development of routine and early screenings for the presence of small RNA species and, therefore, gene silencing in transgenic plants. We further demonstrate that the small RNA fraction can be visualized with the SYBR Green II RNA stain, isolated from a gel, labeled and used as a specific probe. Using these approaches, we have fine-mapped the sequences of the GUS gene that are represented in the small RNA fraction of a GUS-silenced tobacco line containing an inverted repeat of the GUS gene. In this tobacco line, the silencing-associated small RNA is a mixture of fragments that cover the 3' two-thirds of the GUS coding region. The 5' coding and the 3' noncoding ends of the GUS mRNA are not represented in the small RNA fraction. The RNA fragments are not likely to be a primary synthesis product of an RNA-dependent RNA polymerase, but rather degradation products from nuclease activity. Surprisingly, RNA isolated from wild-type, untransformed plants showed the presence of a similar-sized small RNA pool. This might indicate the existence of small RNA species from putative endogenous genes that are down regulated by a similar posttranscriptional gene-silencing mechanism. The possibility of isolating and labeling the small RNA pool from wild-type plants will provide a way to identify and study such putative genes.

Hutvagner, G; Mlynarova, L; Nap, J P



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



Identification and Cloning of gusA, Encoding a New  Glucuronidase from Lactobacillus gasseri ADH  

Microsoft Academic Search

The gusA gene, encoding a new b-glucuronidase enzyme, has been cloned from Lactobacillus gasseri ADH. This is the first report of a b-glucuronidase gene cloned from a bacterial source other than Escherichia coli .A plasmid library of L. gasseri chromosomal DNA was screened for complementation of an E. coli gus mutant. Two overlapping clones that restored b-glucuronidase activity in the

W. M. Russell; T. R. Klaenhammer



Surveys show support for green 'activities'.  


Two independently conducted surveys on sustainability - one into the 'views and values' of NHS 'leaders', and the other questioning the public about the importance of the 'green agenda' in the NHS, and their opinions on how the service might most effectively reduce its carbon footprint, form the basis of Sustainability in the NHS: Health Check 2012, a new NHS Sustainable Development Unit (NHS SDU) publication. As HEJ editor Jonathan Baillie reports, the new document also presents updated data on the 'size' of the carbon footprint of the NHS in England, showing that, although good work by a number of Trusts in the past two years has seen healthcare-generated carbon emissions start to 'level off', the biggest contributors have been the current health service spending review, and the increased national availability of renewable energy. PMID:22515017

Baillie, Jonathan



Promoter analysis in transient assays using a GUS reporter gene construct in creeping bentgrass (Agrostis palustris).  


Transient expression profiles for several chimeric beta-glucuronidase (GUS) gene constructs were determined in tissues (young leaves, mature leaves and roots) of creeping bentgrass (Agrostis palustris, cv. Penn A4) following microprojectile bombardment. The constructs analyzed consisted of the uidA (GUS) reporter gene driven by four different promoters (ubiquitin 3-potato, ubiquitin corn, ubiquitin rice and CaMV 35S). The total number of GUS hits (or transient expression units; TEUs) were determined manually under a dissecting scope after histochemical staining for GUS. Results suggest that the ubiquitin rice promoter is most active in cells of turfgrass, regardless of the developmental stage or tissue-type. The ubiquitin corn promoter was the next best. Of the four promoter used, except for ubiquitin 3-potato, reporter gene activity was dramatically higher in mature leaves compared to young leaves. The relative efficiency of each promoter was about the same in roots and leaves. We have also analyzed uidA (GUS) reporter gene activity following microprojectile bombardment in transient expression assays with callus from two cultivars (Providence or Penn A4) of creeping bentgrass. Differences in the frequency of GUS positive hits were observed between cultivars up to 72 hours post-bombardment. However, this difference between cultivars disappeared after 72 hours post-bombardment. This information describing promoter functionality in bentgrass will be important when designing gene constructs for trait modification and when choosing appropriate cultivars for improvement through gene transfer experiments. This is the first in depth report on organ-specific and developmental gene expression profiles for transgenes in a turfgrass species. PMID:14610892

Basu, Chhandak; Kausch, Albert P; Luo, Hong; Chandlee, Joel M



Transferring Gus gene into intact rice cells by low energy ion beam  

NASA Astrophysics Data System (ADS)

A new technique of transferring genes by low energy ion beam has been reported in this paper. The Gus and CAT (chloramphenicol acetyltransferase) genes, as ``foreign'' genetic materials, were introduced into the suspension cells and ripe embryos or rice by implantation of 20-30 keV Ar+ at doses ranging from 1 × 1015 to 4 × 1015 ions/cm2. The activities of CAT and Gus were detected in the cells and embryos after several weeks. The results indicate that the transfer was a success.

Zengliang, Yu; Jianbo, Yang; Yuejin, Wu; Beijiu, Cheng; Jianjun, He; Yuping, Huo



Biosafety of E. coli ß-glucuronidase (GUS) in plants  

Microsoft Academic Search

The beta-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.

L. J. W. Gilissen; P. L. J. Metz; W. J. Stiekema; J. P. H. Nap



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



Activation and developmental regulation of an Arabidopsis anther-specific promoter in microspores and pollen of Nicotiana tabacum  

Microsoft Academic Search

Seven cytologically distinct stages during micro spore development were identified and used to define the activation of an Arabidopsis anther-specific gene (apg) in transgenic tobacco plants containing an apg promoter-gus fusion. Histochemical analysis of GUS activity showed that the apg promoter was activated in miduninucleate microspores prior to equatorial and polar nuclear migration. Quantitative analysis in isolated spores showed that

David Twell; Samita Patel; Anna Sorensen; Michael Roberts; Rod Scott; John Draper; Gary Foster



Jasmonate-inducible expression of a potato cathepsin D inhibitor-GUS gene fusion in tobacco cells  

Microsoft Academic Search

A potato gene encoding cathepsin D inhibitor (CDI) is expressed constitutively in tubers and flower buds and it is inducible in leaves upon wounding of the tissue or by treatment with methyl jasmonate (MJA). A fusion gene (CDI:GUS) in which the 2.4 kb long promoter of the CDI gene was translationaly fused with the coding sequence for ß-glucuronidase (GUS) showed

Atsushi Ishikawa; Teruhiko Yoshihara; Kenzo Nakamura



Study on transient expression of gus gene in Chlorelia ellipsoidea (Chlorophyta) by using biolistic particle delivery system  

NASA Astrophysics Data System (ADS)

Study on the transient expression of GUS gene at different growing stage of Chlorella ellipsoidea using high velocity microprojectiles, the effects of osmosis, the distance between microprojectile and target cell, bombardment times, are reported in this paper. The results showed that C. ellipsoidea in exponential phase has higer level of transient expression and that treatment with osmosis can improve the GUS transient expression notably. The effect of distance or bombardment times was not observed.

Chen, Ying; Li, Wen-Bin; Bai, Qin-Hua; Sun, Yong-Ru



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.



Characterization of alkaline phosphatase labeled UidA(Gus) probe and its application in testing of transgenic tritordeum.  


Hybridization is a very important molecular biology technique to measure the degree of genetic similarity between DNA sequences, and detect the foreign genes in transgenic organisms. To label a DNA or RNA probe plays a key role in hybridization. A method using nonradioactive material alkaline phosphatase to label UidA(Gus) DNA as probe has been studied. On that basis of Renz and our previous work, alkaline phosphatase-labeled DNA was used as a probe to examine the transformation of the foreign UidA(Gus) gene in transgenic tritordeum. Such DNA-enzyme complexes were characterized and examined carefully, the results showed that it was a sensitive, specific, safe and economical probe. For dot hybridization and Southern blot under full-stringency conditions with alkaline phosphatase as the detector and 5-bromo-4-chloro-3-indolyl phosphate (BCIP)-Nitro Blue Tetrazolium (NBT) as the substrate, dot hybridization showed that the UidA(Gus) gene was transformed into the target plants and inherited stable, Southern blot showed that at least two copies of UidA(Gus) gene were inserted into one line of our transgenic tritordeum. Histochemical staining with X-Gluc of transgenic tritordeum also certified that the foreign UidA(Gus) DNA were transformed into the transgenic tritordeum. PMID:21153926

Tu, Zhiming; Zhang, Jiangzhou; Yang, Guangxiao; He, Guangyuan



A Vigna radiata 8S globulin ?' promoter drives efficient expression of GUS in Arabidopsis cotyledonary embryos.  


Plants are proven effective bioreactors for the production of heterologous proteins including those desired by the biopharmaceutical industry. However, the potential of plants as bioreactors is limited by the availability of characterized plant promoters that can drive target gene expression in relatively distant plant species. Seeds are ideal for protein storage because seed proteins can be kept stably for several months. Hence, a strong promoter that can direct the expression and accumulation of target proteins within seeds represents a powerful tool in plant biotechnology. Toward this end, an effort was made to identify such a promoter from Vigna radiata (mung bean) to drive expression in dicot seeds. A 784-bp 5'-flanking sequence of the gene encoding the 8S globulin ?' subunit (8SG?') of the V. radiata seed storage protein was isolated by genome walking. When the 5'-flanking region was analyzed with bioinformatics tools, numerous putative cis-elements were identified. The Green Fluorescent Protein (GFP) regulated by this promoter was observed to be transiently expressed in protoplasts derived from V. radiata cotyledons. Finally, transgenic Arabidopsis plants expressing the ?-glucuronidase (GUS) reporter gene driven from the 8S globulin ?' promoter showed strong GUS expression in transgenic embryos in both histochemical and quantitative GUS assays, confirming high expression within seeds. Therefore, the V. radiata 8S ?' promoter has shown potential in directing expression in seeds for bioreactor applications. PMID:23763701

Chen, Mo-Xian; Yang, Yue-Ning; Zheng, Shu-Xiao; Xu, Chao; Wang, Ying; Liu, Jie-Sheng; Yang, Wei-Dong; Chye, Mee-Len; Li, Hong-Ye




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



Breaking-off tissue specific activity of the oil palm metallothionein-like gene promoter in T(1) seedlings of tomato exposed to metal ions.  


Metallothioneins (MTs) are cysteine-rich metal-binding proteins that are involved in cell growth regulation, transportation of metal ions and detoxification of heavy metals. A mesocarp-specific metallothionein-like gene (MT3-A) promoter was isolated from the oil palm (Elaeis guineensis Jacq). A vector construct containing the MT3-A promoter fused to the ?-glucuronidase (GUS) gene in the pCAMBIA 1304 vector was produced and used in Agrobacterium-mediated transformation of tomato. Histochemical GUS assay of different tissues of transgenic tomato showed that the MT3-A promoter only drove GUS expression in the reproductive tissues and organs, including the anther, fruit and seed coat. Competitive RT-PCR and GUS fluorometric assay showed changes in the level of GUS mRNA and enzyme activity in the transgenic tomato (T(0)). No GUS mRNA was found in roots and leaves of transgenic tomato. In contrast, the leaves of transgenic tomato seedlings (T(1)) produced the highest GUS activity when treated with 150 ?M Cu(2+) compared to the control (without Cu(2+)). However, Zn(2+) and Fe(2+) treatments did not show GUS expression in the leaves of the transgenic tomato seedlings. Interestingly, the results showed a breaking-off tissue-specific activity of the oil palm MT3-A promoter in T(1) seedlings of tomato when subjected to Cu(2+) ions. PMID:23290536

Kamaladini, Hossein; Nor Akmar Abdullah, Siti; Aziz, Maheran Abdul; Ismail, Ismanizan Bin; Haddadi, Fatemeh



Synthesis and evaluation of dioleoyl glyceric acids showing antitrypsin activity.  


Previously, Lešová et al. reported the isolation and identification of metabolite OR-1, showing antitrypsin activity, produced during fermentation by Penicillium funiculosum. The structure of OR-1 was a mixture of glyceric acid (GA), esterified with C(14)-C(18) fatty acids, and oleic acid (C18:1) as the most predominant fatty acid (Folia Microbiol. 46, 21-23, 2001). In this study, dioleoyl D-GA and dioleoyl L-GA were synthesized via diesterification with oleoyl chloride, and their antitrypsin activities were evaluated using both a disk diffusion method and spectral absorption measurements. The results show that both compounds and their equivalent mixtures possess antitrypsin activities; however, their IC(50) values (approximately 2 mM) are much higher than that of OR-1 (4.25 µM), suggesting that dioleoyl GA does not play a major role in the OR-1 antitrypsin activity. PMID:21606621

Habe, Hiroshi; Fukuoka, Tokuma; Sato, Shun; Kitamoto, Dai; Sakaki, Keiji



Transformation of Rhododendron spp. using Agrobacterium tumefaciens with a GUS-intron chimeric gene  

Microsoft Academic Search

The five Rhododendron cultivars, ‘America’, ‘Catawbiense grandiflorum roseum’, ‘Madame Carvalho’, ‘Mars’ and ‘Nova Zembla’ were used for transformation by Agrobacterium tumefaciens carrying T-DNA with the gusA gene encoding ?-glucuronidase (GUS) gene and the neomycin phosphotransferase II gene as a selectable marker gene. The GUS reporter gene was successfully transferred into all five cultivars as indicated by fluorimetric staining, polymerase chain

Daniela Pavingerová; Jind?ich Bríza; Karel Kodýtek; Hana Niedermeierová



NASA's Fermi Shows How Active Galaxies Can Be  

NASA Video Gallery

Active galaxies called blazars make up the largest class of objects detected by Fermi's Large Area Telescope (LAT). Massive black holes in the hearts of these galaxies fire particle jets in our direction. Fermi team member Elizabeth Hays narrates this quick tour of blazars, which includes LAT movies showing how rapidly their emissions can change. Credit: NASA/Goddard Space Flight Center




Analysis of genetically modified plant gene expression using GUS fluorimetry  

Microsoft Academic Search

A fluorimetric assay method for the analysis of beta-glucuronidase (GUS) reporter gene expression in genetically modified\\u000a plants is described. Optimization of this method for woody plants and a statistical approach suitable for comparisons of gene\\u000a expression in different transformants or tissues of the same plant is described. Example data from elm (Ulmus procera) SR4 regenerant plants, shown to be genetically

Kevan M. A. Gartland; Angela T. McHugh; Stanislav Vitha; Karel Benes; Richard J. Irvine; Jill S. Gartland



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


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 Mourăo Filho, Francisco



Synthetic analogs of anoplin show improved antimicrobial activities.  


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. Copyright © 2013 European Peptide Society and John Wiley & Sons, Ltd. PMID:24019229

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



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



Thalidomide Shows Activity Against AIDS-Related Kaposi's Sarcoma

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).


?-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



Australian plants show anthelmintic activity toward equine cyathostomins in vitro.  


Anthelmintic resistance in gastrointestinal parasites of horses is an increasing problem, particularly in cyathostomins, and there is a need to find alternative means for the control of these parasites. We screened crude extracts from 37 species of Australian native plants for their anthelmintic activity in vitro against cyathostomin larvae (development from egg to third larval stage), with the aim of identifying those species that may be suitable for incorporation into sustainable parasite management programs. Water extracts from seven species, namely Acacia baileyana, Acacia melanoxylon, Acacia podalyriifolia, Alectryon oleifolius, Duboisia hopwoodii, Eucalyptus gomphocephala and Santalum spicatum completely inhibited larval development (100% inhibition compared to the control), while another 10 species caused 90% inhibition at the initial screening concentration of 1400 ?g of extractable solids/mL. The seven most potent extracts produced IC50 values (concentration of extract which resulted in a 50% inhibition of development) in the range 30.9-196 ?g/mL. Fourteen extracts were incubated with polyvinylpolypyrrolidone (PVPP) before the assays, which removed the anthelmintic activity from 12 of these extracts, indicating that tannins were likely to be the bioactive compound responsible for the effect, while in two species, i.e. A. melanoxylon and D. hopwoodii, compounds other than tannins were likely to be responsible for their anthelmintic action. Our results suggest that a number of Australian native plants have significant anthelmintic activity against cyathostomin larval development in vitro. There is potential for these plants to be used as part of sustainable parasite control programs in horses, although more research is needed to identify the compounds responsible for the anthelmintic effects and confirm their activity in vivo. PMID:23394801

Payne, S E; Kotze, A C; Durmic, Z; Vercoe, P E



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.



A novel nucleic acid analogue shows strong angiogenic activity.  


A novel nucleic acid analogue (2Cl-C.OXT-A) significantly stimulated tube formation of human umbilical endothelial cells (HUVEC). Its maximum potency at 100muM was stronger than that of vascular endothelial growth factor (VEGF), a positive control. At this concentration, 2Cl-C.OXT-A moderately stimulated proliferation as well as migration of HUVEC. To gain mechanistic insights how 2Cl-C.OXT-A promotes angiogenic responses in HUVEC, we performed immunoblot analyses using phospho-specific antibodies as probes. 2Cl-C.OXT-A induced robust phosphorylation/activation of MAP kinase ERK1/2 and an upstream MAP kinase kinase MEK. Conversely, a MEK inhibitor PD98059 abolished ERK1/2 activation and tube formation both enhanced by 2Cl-C.OXT-A. In contrast, MAP kinase responses elicited by 2Cl-C.OXT-A were not inhibited by SU5416, a specific inhibitor of VEGF receptor tyrosine kinase. Collectively these results suggest that 2Cl-C.OXT-A-induces angiogenic responses in HUVEC mediated by a MAP kinase cascade comprising MEK and ERK1/2, but independently of VEGF receptor tyrosine kinase. In vivo assay using chicken chorioallantoic membrane (CAM) and rabbit cornea also suggested the angiogenic potency of 2Cl-C.OXT-A. PMID:20691660

Tsukamoto, Ikuko; Sakakibara, Norikazu; Maruyama, Tokumi; Igarashi, Junsuke; Kosaka, Hiroaki; Kubota, Yasuo; Tokuda, Masaaki; Ashino, Hiromi; Hattori, Kenichi; Tanaka, Shinji; Kawata, Mitsuhiro; Konishi, Ryoji



Adults' preferences between Picture Communication Symbols (PCSs) and Gus Communication Symbols (GCSs) used in AAC.  


The purpose of the study was to determine whether significantly different preference exists in typical adults between Picture Communication Symbols (PCSs) and Gus Communication Symbols (GCSs) frequently displayed on their AAC. A total of 56 participants participated in the study, including 15 participants in ages 20-35; 14 participants in ages 36-50; 13 participants in ages 51-60; and 14 participants in ages 66-80. Two tasks, an identification task and a preference task, were administered sequentially and individually. The results of this study suggest: (1) typical adults' preference between PCSs and GCSs did not show any significant difference; (2) the effect of age groups of typical adults did not have any significant effect on their preference between PCSs and GCSs. However, except the group of ages 51-65, the other three groups did consciously show their preferences for PCSs. Limitations and clinical implications of the current study have been concluded. PMID:23962601

Tsai, Meng-Ju



Technical gains show big improvement in marine seismic activity  

SciTech Connect

Continued economic recovery in the US and abroad will be necessary to increase demand for petroleum products and help stabilize current price fluctuations. Price stability should be achieved during 1985 and the result will be a continuation of the recovery currently underway in the offshore industry. Political and economic developments will have an important effect on the decisions that are made in the offshore industry during 1985. Offshore oil and gas exploration and development in the US will be greatly affected by policy decisions that are made on the current offshore leasing program. During the past two years, large amounts of acreage have been withdrawn from the program in response to various concerns and recommendations expressed by coastal states, governors and others. Seismic exploration continues as a leading indicator of future oil and gas exploration activity. Seismic crew counts in the US and internationally have been improving since bottoming out in early 1983.

Heitman, L.B.



Transient expression of GUS and the 2S albumin gene from Brazil nut in peanut ( Arachis hypogaea L.) seed explants using particle bombardment  

Microsoft Academic Search

The effect of parameters involved in the transformation efficiency of peanut (Arachis hypogaea L.) seed tissues by direct gene transfer using a helium inflow particle bombardment device was evaluated. Transient gene expression was affected by both particle and DNA amounts, and was positively correlated with gene copy number, as determined byß-glucuronidase (GUS) activity assays. No influence of plasmid size on

C. Lacorte; F. J. L. Aragăo; E. R. Almeida; E. L. Rech; E. Mansur



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.




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



GUS expression in blueberry (Vaccinium spp.): factors influencing Agrobacterium-mediated gene transfer efficiency  

Microsoft Academic Search

Several factors were investigated for their influence on the transfer of an intron-containing ?-glucuronidase (GUS) gene into blueberry (Vaccinium spp.) leaf explants during the early stages of Agrobacterium-mediated gene transfer, including days of cocultivation, strain of Agrobacterium tumefaciens, explant age and genotype. The number of GUS-expressing leaf zones and calli were counted immediately and 2 weeks after cocultivation,\\u000a respectively, to

X. Cao; Q. Liu; L. J. Rowland; F. A. Hammerschlag



Specific degradation of 3' regions of GUS mRNA in posttranscriptionally silenced tobacco lines may be related to 5'-3' spreading of silencing.  

PubMed Central

Target regions for posttranscriptional silencing of transgenes often reside in the 3' region of the coding sequence, although there are exceptions. To resolve if the target region is determined by the gene undergoing silencing rather than by the structure of the transgene loci or the plant genetic background, we have performed detailed analyses of target regions in three spontaneously beta-glucuronidase (GUS) silencing tobacco lines of different origin. From quantitative cosuppression experiments, we show that the main target region in all three tobacco lines is found within the 3' half of the GUS coding region but upstream of the last 200 nt. The quantities of small (21-25 nt) RNAs homologous to 5' or 3' regions of the GUS coding sequence were found to correlate approximately with the target strength of the corresponding regions. These results suggest that transgene locus structure and plant genetic background are not major determinants of silencing target regions. We also show that virus-induced gene silencing (VIGS) of GUS in Nicotiana benthamiana is induced equally effectively with Potato virus X carrying either the 5' or 3' third of the GUS coding region. This indicates that both regions can act as efficient inducers as well as targets of posttranscriptional silencing, although the 3' region is the predominant target region in the spontaneously silencing transgenic plant lines examined. Finally, we investigated spreading of the target region in the N. benthamiana plants undergoing VIGS. Surprisingly, only evidence for spreading of the target region in the 5'-3' direction was obtained. This finding may help explain why the majority of target regions examined to date lie within the 3' region of transgenes.

Braunstein, Thomas Hartig; Moury, Benoit; Johannessen, Marina; Albrechtsen, Merete



A Paradox of Syntactic Priming: Why Response Tendencies Show Priming for Passives, and Response Latencies Show Priming for Actives  

PubMed Central

Speakers tend to repeat syntactic structures across sentences, a phenomenon called syntactic priming. Although it has been suggested that repeating syntactic structures should result in speeded responses, previous research has focused on effects in response tendencies. We investigated syntactic priming effects simultaneously in response tendencies and response latencies for active and passive transitive sentences in a picture description task. In Experiment 1, there were priming effects in response tendencies for passives and in response latencies for actives. However, when participants' pre-existing preference for actives was altered in Experiment 2, syntactic priming occurred for both actives and passives in response tendencies as well as in response latencies. This is the first investigation of the effects of structure frequency on both response tendencies and latencies in syntactic priming. We discuss the implications of these data for current theories of syntactic processing.

Segaert, Katrien; Menenti, Laura; Weber, Kirsten; Hagoort, Peter



Visualization of auxin-mediated transcriptional activation using a common auxin-responsive reporter system in the liverwort Marchantia polymorpha.  


The phytohormone auxin plays a pivotal role in various developmental aspects in land plants. However, little is known of the auxin response and distribution in non-vascular plants. In this study, we made transgenic plants of the liverwort Marchantia polymorpha which express the uidA (GUS) reporter gene under control of the soybean auxin-inducible promoter, ProGH3, and used it to indirectly monitor auxin-mediated transcriptional activation in planta. Transgenic plants carrying ProGH3:GUS showed GUS activity in an auxin-dependent manner. Histochemical GUS staining was observed at the bottom of gemma cups in the process of vegetative propagation. Significant GUS activity was also detected around the gametophyte-sporophyte junction as well as the developing sporophyte after fertilization. These results suggest that the activity of auxin is crucial in both gametophyte and sporophyte development in M. polymorpha, and that the mechanism for auxin-mediated transcriptional activation had already been established when plants emerged on the terrestrial environment. PMID:22311005

Ishizaki, Kimitsune; Nonomura, Maiko; Kato, Hirotaka; Yamato, Katsuyuki T; Kohchi, Takayuki



An Apologetic to the Neo-Pagans as Represented by Dr. Gus DiZerega  

Microsoft Academic Search

The current trend of multi-religious observance can be traced to various root sources. In particular, the advocacy of Christianity and another religious tradition is particularly strong within Neo-Pagan circles. Through historical investigation, as well as response to the writings of Dr. Gus DiZerega, a prominent Neo-Pagan proponent, these connection will be made. It will be posited that the core of

William Tennison Smitherman II



Chrysanthemum cultivar– Agrobacterium interactions revealed by GUS expression time course experiments  

Microsoft Academic Search

Nine 24-day GUS expression time course experiments involving 2760 leaf explants revealed complex host–bacterium interactions between four cultivars of florists' chrysanthemum from two sources (in vitro and glasshouse) and four disarmed strains of Agrobacterium tumefaciens (EHA105, LBA4404, MOG101, MOG301) containing the binary vectors pMOG410 or pKIWI110. Cultivars Peach Margaret and Yellow Lucondra [DendranthemaGrandiflorum] were more easily transformed than cv. Korean

M. R Boase; R. C Butler; N. K Borst



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


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



Glyceraldehyde-3-phosphate dehydrogenase from chironomidae showed differential activity towards metals.  


Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is known to interact with different biomolecules and was implicated in many novel cellular activities including programmed cell death, nuclear RNA transport unrelated to the commonly known carbohydrate metabolism. We reported here the purification of GAPDH from Chironomidae larvae (Insecta, Diptera) that showed different biologic activity towards heavy metals. It was inhibited by copper, cobalt nickel, iron and lead but was activated by zinc. The GAPDH was purified by ammonium sulphate fractionation and Chelating Sepharose CL-6B chromatography followed by Blue Sepharose CL-6B chromatography. The 150-kDa tetrameric GAPDH showed optimal activity at pH 8.5 and 37°C. The multiple alignment of sequence of the Chironomidae GAPDH with other known species showed 78 - 88% identity to the conserved regions of the GADPH. Bioinformatic analysis unveils substantial N-terminal sequence similarity of GAPDH of Chironomidae larvae to mammalian GADPHs. However, the GADPH of Chironomidae larvae showed different biologic activities and cytotoxicity towards heavy metals. The GAPDH enzyme would undergo adaptive molecular changes through binding at the active site leading to higher tolerance to heavy metals. PMID:23607280

Chong, Isaac K W; Ho, Wing S



Genetic transformation of potato with nptII-gus marker genes enhances foliage consumption by Colorado potato beetle larvae  

Microsoft Academic Search

Little is known about the effect of transgenic plants containing commonly used marker genes, such as aph(3')II (nptII encoding neomycinphosphotransferase) and uidA (gus encoding ß-glucuronidase) on insect feeding behaviour. We report here, for the first time, that transgenic potato plants containing only nptII and gus marker genes enhance foliage consumption by the Colorado potato beetle (CPB, Leptinotarsa decemlineata S.). Transformation

Anne Lecardonnel; Genevičve Prévost; Antony Beaujean; Rajbir S. Sangwan; Brigitte S. Sangwan-Norreel



Endogenous Dipeptide Cycloprolylglycine Shows Selective Anxiolytic Activity in Animals with Manifest Fear Reaction  

Microsoft Academic Search

Experiments on two mouse strains with opposite reactions to emotional stress showed selectivity of the anxiolytic effect of endogenous dipeptide cycloprolylglycine. In the open field test cycloprolylglycine (0.01-0.10 mg\\/kg intraperitoneally) dose-dependently (1.8-2.1-fold) increased motor activity of BALB\\/c mice with manifest fear reaction and had no effect on C57Bl\\/6 mice with active behavior. The content of endogenous cycloprolylglycine in mouse brain

S. B. Seredenin; T. A. Gudasheva; S. S. Boiko; G. I. Kovalev; M. V. Voronin; M. A. Yarkova



A humanized anti-M2 scFv shows protective in vitro activity against influenza  

SciTech Connect

M2 is one of the most conserved influenza proteins, and has been widely prospected as a potential universal vaccine target, with protection predominantly mediated by antibodies. In this paper we describe the creation of a humanized single chain Fv from 14C2, a potent monoclonal antibody against M2. We show that the humanized scFv demonstrates similar activity to the parental mAb: it is able to recognize M2 in its native context on cell surfaces and is able to show protective in vitro activity against influenza, and so represents a potential lead antibody candidate for universal prophylactic or therapeutic intervention in influenza.

Bradbury, Andrew M [Los Alamos National Laboratory; Velappan, Nileena [Los Alamos National Laboratory; Schmidt, Jurgen G [Los Alamos National Laboratory



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



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

PubMed Central

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 designated Bmlipase-1. Northern blot analysis showed that the Bmlipase-1 gene is expressed in the midgut but not in other tissues, nor is it activated by BmNPV infection. In addition, the Bmlipase-1 gene was shown not to be expressed in the molting and wandering stages, indicating that the gene is hormonally regulated. Our results suggest that an insect digestive enzyme has potential as a physiological barrier against BmNPV at the initial site of viral infection.

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



Psychoacoustic Tinnitus Loudness and Tinnitus-Related Distress Show Different Associations with Oscillatory Brain Activity  

PubMed Central

Background The phantom auditory perception of subjective tinnitus is associated with aberrant brain activity as evidenced by magneto- and electroencephalographic studies. We tested the hypotheses (1) that psychoacoustically measured tinnitus loudness is related to gamma oscillatory band power, and (2) that tinnitus loudness and tinnitus-related distress are related to distinct brain activity patterns as suggested by the distinction between loudness and distress experienced by tinnitus patients. Furthermore, we explored (3) how hearing impairment, minimum masking level, and (4) psychological comorbidities are related to spontaneous oscillatory brain activity in tinnitus patients. Methods and Findings Resting state oscillatory brain activity recorded electroencephalographically from 46 male tinnitus patients showed a positive correlation between gamma band oscillations and psychoacoustic tinnitus loudness determined with the reconstructed tinnitus sound, but not with the other psychoacoustic loudness measures that were used. Tinnitus-related distress did also correlate with delta band activity, but at electrode positions different from those associated with tinnitus loudness. Furthermore, highly distressed tinnitus patients exhibited a higher level of theta band activity. Moreover, mean hearing loss between 0.125 kHz and 16 kHz was associated with a decrease in gamma activity, whereas minimum masking levels correlated positively with delta band power. In contrast, psychological comorbidities did not express significant correlations with oscillatory brain activity. Conclusion Different clinically relevant tinnitus characteristics show distinctive associations with spontaneous brain oscillatory power. Results support hypothesis (1), but exclusively for the tinnitus loudness derived from matching to the reconstructed tinnitus sound. This suggests to preferably use the reconstructed tinnitus spectrum to determine psychoacoustic tinnitus loudness. Results also support hypothesis (2). Moreover, hearing loss and minimum masking level correlate with oscillatory power in distinctive frequency bands. The lack of an association between psychological comorbidities and oscillatory power may be attributed to the overall low level of mental health problems in the present sample.

Balkenhol, Tobias; Wallhausser-Franke, Elisabeth; Delb, Wolfgang



A cry in the dark: depressed mothers show reduced neural activation to their own infant's cry  

PubMed Central

This study investigated depression-related differences in primiparous mothers’ neural response to their own infant’s distress cues. Mothers diagnosed with major depressive disorder (n?=?11) and comparison mothers with no diagnosable psychopathology (n?=?11) were exposed to their own 18-months-old infant’s cry sound, as well as unfamiliar infant’s cry and control sound, during functional neuroimaging. Depressed mothers’ response to own infant cry greater than other sounds was compared to non-depressed mothers’ response in the whole brain [false discovery rate (FDR) corrected]. A continuous measure of self-reported depressive symptoms (CESD) was also tested as a predictor of maternal response. Non-depressed mothers activated to their own infant’s cry greater than control sound in a distributed network of para/limbic and prefrontal regions, whereas depressed mothers as a group failed to show activation. Non-depressed compared to depressed mothers showed significantly greater striatal (caudate, nucleus accumbens) and medial thalamic activation. Additionally, mothers with lower depressive symptoms activated more strongly in left orbitofrontal, dorsal anterior cingulate and medial superior frontal regions. Non-depressed compared to depressed mothers activated uniquely to own infant greater than other infant cry in occipital fusiform areas. Disturbance of these neural networks involved in emotional response and regulation may help to explain parenting deficits in depressed mothers.

Ablow, Jennifer C.



A cry in the dark: depressed mothers show reduced neural activation to their own infant's cry.  


This study investigated depression-related differences in primiparous mothers' neural response to their own infant's distress cues. Mothers diagnosed with major depressive disorder (n?=?11) and comparison mothers with no diagnosable psychopathology (n?=?11) were exposed to their own 18-months-old infant's cry sound, as well as unfamiliar infant's cry and control sound, during functional neuroimaging. Depressed mothers' response to own infant cry greater than other sounds was compared to non-depressed mothers' response in the whole brain [false discovery rate (FDR) corrected]. A continuous measure of self-reported depressive symptoms (CESD) was also tested as a predictor of maternal response. Non-depressed mothers activated to their own infant's cry greater than control sound in a distributed network of para/limbic and prefrontal regions, whereas depressed mothers as a group failed to show activation. Non-depressed compared to depressed mothers showed significantly greater striatal (caudate, nucleus accumbens) and medial thalamic activation. Additionally, mothers with lower depressive symptoms activated more strongly in left orbitofrontal, dorsal anterior cingulate and medial superior frontal regions. Non-depressed compared to depressed mothers activated uniquely to own infant greater than other infant cry in occipital fusiform areas. Disturbance of these neural networks involved in emotional response and regulation may help to explain parenting deficits in depressed mothers. PMID:21208990

Laurent, Heidemarie K; Ablow, Jennifer C



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

Microsoft Academic Search

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 Ni3(shi)2(Hpko)2(py)2(1), two metal ions show a square planar geometry while the third one is in an

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



Meropenem-Clavulanic Acid Shows Activity against Mycobacterium tuberculosis In Vivo  

PubMed Central

The carbapenems imipenem and meropenem in combination with clavulanic acid reduced the bacterial burden in Mycobacterium tuberculosis-infected macrophages by 2 logs over 6 days. Despite poor stability in solution and a short half-life in rodents, treatment of chronically infected mice revealed significant reductions of bacterial burden in the lungs and spleens. Our results show that meropenem has activity in two in vivo systems, but stability and pharmacokinetics of long-term administration will offer significant challenges to clinical evaluation.

England, Kathleen; Boshoff, Helena I. M.; Arora, Kriti; Weiner, Danielle; Dayao, Emmanuel; Schimel, Daniel; Via, Laura E.



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


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



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

PubMed Central

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 broad spectrum of antimicrobial activities. Their potent anti-pathogenic characteristics have ensured that they are promising targets in the medical and agricultural biotechnology sectors. Results A berry specific cDNA sequence designated Vv-AMP1, Vitis vinifera antimicrobial peptide 1, was isolated from Vitis vinifera. Vv-AMP1 encodes for a 77 amino acid peptide that shows sequence homology to the family of plant defensins. Vv-AMP1 is expressed in a tissue specific, developmentally regulated manner, being only expressed in berry tissue at the onset of berry ripening and onwards. Treatment of leaf and berry tissue with biotic or abiotic factors did not lead to increased expression of Vv-AMP1 under the conditions tested. The predicted signal peptide of Vv-AMP1, fused to the green fluorescent protein (GFP), showed that the signal peptide allowed accumulation of its product in the apoplast. Vv-AMP1 peptide, produced in Escherichia coli, had a molecular mass of 5.495 kDa as determined by mass spectrometry. Recombinant Vv-AMP1 was extremely heat-stable and showed strong antifungal activity against a broad spectrum of plant pathogenic fungi, with very high levels of activity against the wilting disease causing pathogens Fusarium oxysporum and Verticillium dahliae. The Vv-AMP1 peptide did not induce morphological changes on the treated fungal hyphae, but instead strongly inhibited hyphal elongation. A propidium iodide uptake assay suggested that the inhibitory activity of Vv-AMP1 might be associated with altering the membrane permeability of the fungal membranes. Conclusion A berry specific cDNA clone, Vv-AMP1, was isolated and characterized and shown to encode a plant defensin. Recombinant Vv-AMP1 displayed non-morphogenic antifungal activity against a broad spectrum of fungi, probably altering the membrane permeability of the fungal pathogens. The expression of this peptide is highly regulated in Vitis vinifera, hinting at an important defense role during berry-ripening.

de Beer, Abre; Vivier, Melane A



High and low sensation seeking adolescents show distinct patterns of brain activity during reward processing.  


Previous research has shown that personality characteristics, such as sensation seeking (SS), are strong predictors of risk-taking behavior during adolescence. However, the relationship between levels of SS and brain response has not been studied during this time period. Given the prevalence of risky behavior during adolescence, it is important to understand neurobiological differences in reward sensitivity between youth with high and low SS personalities. To this end, we used functional magnetic resonance imaging (fMRI) to examine differences in brain activity in an adolescent sample that included 27 high (HSS) and 27 low sensation seekers (LSS), defined by the Impulsive Sensation Seeking scale of the Zuckerman-Kuhlman Personality Questionnaire (Zuckerman et al., 1993). In the scanner, participants played a modified Wheel of Fortune decision-making task (Cservenka and Nagel, 2012) that resulted in trials with monetary Wins or No Wins. We compared age- and sex-matched adolescent HSS and LSS (mean age=13.94±1.05) on brain activity by contrasting Win vs. No Win trials. Our findings indicate that HSS show greater bilateral insular and prefrontal cortex (PFC) brain response on Win vs. No Win compared to LSS. Analysis of simple effects showed that while LSS showed comparable brain activity in these areas during Wins and No Wins, HSS showed significant differences in brain response to winning (activation) vs. not winning (deactivation), with between-group comparison suggesting significant differences in brain response, largely to reward absence. Group differences in insular activation between reward receipt and absence may suggest weak autonomic arousal to negative outcomes in HSS compared with LSS. Additionally, since the PFC is important for goal-directed behavior and attention, the current results may reflect that HSS allocate fewer attentional resources to negative outcomes than LSS. This insensitivity to reward absence in HSS may lead to a greater likelihood of maladaptive choices when negative consequences are not considered, and may be an early neural marker of decreased loss sensitivity that has been seen in addiction. This neurobiological information may ultimately be helpful in establishing prevention strategies aimed at reducing youth risk-taking and suggests value in further examination of neural associations with personality characteristics during adolescence. PMID:23142276

Cservenka, Anita; Herting, Megan M; Seghete, Kristen L Mackiewicz; Hudson, Karen A; Nagel, Bonnie J



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.



Plant vermicides of Haitian Vodou show in vitro activity against larval hookworm.  


Haitian Vodou priests (houngans) and priestesses (mambos) use plant remedies to treat many illnesses, including intestinal parasite infections. The present study screened 12 plants used in Vodou treatments for intestinal parasites to detect in vitro activity against infective-stage larvae of the hookworm Ancylostoma caninum. Water-soluble extracts of 4 of the 12 plants inhibited serum-stimulated feeding by larval A. caninum in a dose-dependent manner. All 4 plant extracts inhibited feeding induced by the muscarinic agonist arecoline, suggesting that these plant extracts may inhibit the insulin-like signaling pathway involved in the recovery and resumption of development of arrested A. caninum larvae. These results indicate that at least some of the plants used in traditional Haitian medicine as vermifuges show activity against nematode physiological processes. PMID:18576795

Wolpert, Beverly J; Beauvoir, Max G; Wells, Elizabeth Fortson; Hawdon, John M



Exposure of erythrocytes to methylene blue shows the active role of catalase in removing hydrogen peroxide.  


Methylene blue (MB) is a powerful reducing agent that is widely used in clinical practice as well as for metabolic studies of the erythrocyte. We have investigated the role of catalase as a specific enzyme for the removal of hydrogen peroxide by measuring the in vitro effects of MB on human red cells. In the presence of MB, catalase underwent inactivation even with the co-existence of active generation of NADPH, leaving the glutathione concentration unaffected. The data obtained in the present investigation show, using a different tool (MB), that catalase is the active enzyme in H2O2 detoxification and that its integrity is largely dependent on an adequate generation of NADPH. PMID:12437668

Gaetani, Gian Franco; Rapezzi, Davide; Mangerini, Rosa; Racchi, Omar; Rolfo, Michela; Ferraris, Anna Maria



Youth at Risk for Obesity Show Greater Activation of Striatal and Somatosensory Regions to Food  

PubMed Central

Obese versus normal-weight humans have less striatal D2 receptors and striatal response to food intake, and weaker striatal response to food predicts weight gain for individuals at genetic risk for reduced dopamine (DA) signaling, consistent with the reward deficit theory of obesity. Yet these may not be initial vulnerability factors, as overeating reduces D2 receptor density, D2 sensitivity, reward sensitivity, and striatal response to food. Obese versus normal-weight humans also show greater striatal, amygdalar, orbitofrontal cortex, and somatosensory region response to food images, which predicts weight gain for those not at genetic risk for compromised dopamine signaling, consonant with the reward surfeit theory of obesity. However, after pairings of palatable food intake and predictive cues, DA signaling increases in response to the cues, implying that eating palatable food contributes to increased responsivity. We tested whether normal-weight adolescents at high- versus low-risk for obesity showed aberrant activation of reward circuitry in response to receipt and anticipated receipt of palatable food and monetary reward using fMRI. High-risk youth showed greater activation in the caudate, parietal operculum, and frontal operculum in response to food intake and in the caudate, putamen, insula, thalamus, and orbitofrontal cortex in response to monetary reward. No differences emerged in response to anticipated food or monetary reward. Data indicate that youth at risk for obesity show elevated reward circuitry responsivity in general coupled with elevated somatosensory region responsivity to food, which may lead to overeating that produces blunted dopamine signaling and elevated responsivity to food cues.

Stice, E; Yokum, S; Burger, KS; Epstein, LH; Small, DM



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.



CpG oligodeoxynucleotides with double stem-loops show strong immunostimulatory activity.  


Based on the current understanding of TLR9 recognition of CpG ODN, we have tried to design a series of CpG ODNs that display double stem-loops when being analyzed for their secondary structures using 'mfold web server'. Proliferation of human PBMC and bioassay for IFN production were used as technical platforms in primary screening. Interestingly, two of them, designated as DSL01 and D-SL03, belonging to B class CpG ODN and C class CpG ODN respectively, showed vigorous immunostimulatory activity and were chosen for further tests. Flow cytometry analysis showed that both of them could activate human B cells, NK cells, mononuclear cells and T cells and up-regulate expression of CD80, CD86 and HLA-DR on the surface of subsets in human PBMCs. Furthermore, we demonstrated that those two ODNs potently stimulated proliferation of PBMC/splenocytes obtained from diverse vertebrate species. Noticeably, both of them displayed anti-breast cancer effect in mice when administered by peritumoral injection. PMID:23142503

Yang, Liang; Wu, Xiuli; Wan, Min; Yu, Yue; Yu, Yongli; Wang, Liying



Ovine PrP transgenic Drosophila show reduced locomotor activity and decreased survival.  


Drosophila have emerged as a model system to study mammalian neurodegenerative diseases. In the present study we have generated Drosophila transgenic for ovine PrP (prion protein) to begin to establish an invertebrate model of ovine prion disease. We generated Drosophila transgenic for polymorphic variants of ovine PrP by PhiC31 site-specific germ-line transformation under expression control by the bi-partite GAL4/UAS (upstream activating sequence) system. Site-specific transgene insertion in the fly genome allowed us to test the hypothesis that single amino acid codon changes in ovine PrP modulate prion protein levels and the phenotype of the fly when expressed in the Drosophila nervous system. The Arg(154) ovine PrP variants showed higher levels of PrP expression in neuronal cell bodies and insoluble PrP conformer than did His(154) variants. High levels of ovine PrP expression in Drosophila were associated with phenotypic effects, including reduced locomotor activity and decreased survival. Significantly, the present study highlights a critical role for helix-1 in the formation of distinct conformers of ovine PrP, since expression of His(154) variants were associated with decreased survival in the absence of high levels of PrP accumulation. Collectively, the present study shows that variants of the ovine PrP are associated with different spontaneous detrimental effects in ovine PrP transgenic Drosophila. PMID:22435640

Thackray, Alana M; Muhammad, Farooq; Zhang, Chang; Di, Ying; Jahn, Thomas R; Landgraf, Matthias; Crowther, Damian C; Evers, Jan Felix; Bujdoso, Raymond



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



Planarity of heteroaryldithiocarbazic acid derivatives showing tuberculostatic activity. IV. Diesters of benzoylcarbonohydrazonodithioic acid.  


Dimethyl (3,4-dichlorobenzoyl)carbonohydrazonodithioate, C(10)H(10)Cl(2)N(2)OS(2), (D1), dibenzyl (3,4-dichlorobenzoyl)carbonohydrazonodithioate, C(22)H(18)Cl(2)N(2)OS(2), (D2), dimethyl (3,4-dichlorobenzoyl)-1-methylcarbonohydrazonodithioate, C(11)H(12)Cl(2)N(2)OS(2), (D3), 3,4-dichloro-N'-(1,3-dithiolan-2-ylidene)-N-methylbenzohydrazide, C(11)H(10)Cl(2)N(2)OS(2), (D4), were synthesized as potential tuberculostatics. Compound (D1) (with two molecules in the asymmetric unit) was the only one showing tuberculostatic activity of the same range as the common drugs isoniazid and pyrazinamide. The molecular structures of the studied compounds depend on the substitution at the N atom adjacent to the carbonyl group. In the case of the unsubstituted derivatives (D1) and (D2), their central frames are generally planar with a twist of the 3,4-dichlorophenyl ring by 30-40°. Until now, coplanarity of the aromatic ring with the (methylene)carbonohydrazone fragment has been considered a prerequisite for tuberculostatic activity. The N-methylated derivatives (D3) and (D4) show an additional twist along the N-C(=O) bond by 20-30° due to the spatial repulsion introduced by the methyl substituent. PMID:22382551

Szczesio, Ma?gorzata; Olczak, Andrzej; Gobis, Katarzyna; Foks, Henryk; G?ówka, Marek L



Inhibition of HIV-1 replication by ribozymes that show poor activity in vitro.  


Self-cleaving RNAs (ribozymes) can be engineered to cleave target RNAs of choice in a sequence-specific manner (1). Consequently, they could be used to inhibit virus replication or to analyse host gene function in vivo. However, ribozymes that are catalytic in vitro are generally disappointing when analysed in cells unless expressed at high levels relative to their target RNAs (2, 3). Here we provide evidence that this can be overcome by optimizing ribozyme structure using cellular rather than cell-free assays. We show that ribozymes of relatively long flanking complementary regions (FCRs), while poor catalysts in vitro, can produce profound inhibition of HIV replication in cells. By examining a series of ribozymes in which the FCRs vary from 9 to 564 nucleotides, we establish that the optimum length for activity in the cell is > or = 33 nucleotides. PMID:8255782

Crisell, P; Thompson, S; James, W



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.



Yarrowia lipolytica Mutants Devoid of Pyruvate Carboxylase Activity Show an Unusual Growth Phenotype†  

PubMed Central

We have cloned and characterized the gene PYC1, encoding the unique pyruvate carboxylase in the dimorphic yeast Yarrowia lipolytica. The protein putatively encoded by the cDNA has a length of 1,192 amino acids and shows around 70% identity with pyruvate carboxylases from other organisms. The corresponding genomic DNA possesses an intron of 269 bp located 133 bp downstream of the starting ATG. In the branch motif of the intron, the sequence CCCTAAC, not previously found at this place in spliceosomal introns of Y. lipolytica, was uncovered. Disruption of the PYC1 gene from Y. lipolytica did not abolish growth in glucose-ammonium medium, as is the case in other eukaryotic microorganisms. This unusual growth phenotype was due to an incomplete glucose repression of the function of the glyoxylate cycle, as shown by the lack of growth in that medium of double pyc1 icl1 mutants lacking both pyruvate carboxylase and isocitrate lyase activity. These mutants grew when glutamate, aspartate, or Casamino Acids were added to the glucose-ammonium medium. The cDNA from the Y. lipolytica PYC1 gene complemented the growth defect of a Saccharomyces cerevisiae pyc1 pyc2 mutant, but introduction of either the S. cerevisiae PYC1 or PYC2 gene into Y. lipolytica did not result in detectable pyruvate carboxylase activity or in growth on glucose-ammonium of a Y. lipolytica pyc1 icl1 double mutant.

Flores, Carmen-Lisset; Gancedo, Carlos



Yarrowia lipolytica mutants devoid of pyruvate carboxylase activity show an unusual growth phenotype.  


We have cloned and characterized the gene PYC1, encoding the unique pyruvate carboxylase in the dimorphic yeast Yarrowia lipolytica. The protein putatively encoded by the cDNA has a length of 1,192 amino acids and shows around 70% identity with pyruvate carboxylases from other organisms. The corresponding genomic DNA possesses an intron of 269 bp located 133 bp downstream of the starting ATG. In the branch motif of the intron, the sequence CCCTAAC, not previously found at this place in spliceosomal introns of Y. lipolytica, was uncovered. Disruption of the PYC1 gene from Y. lipolytica did not abolish growth in glucose-ammonium medium, as is the case in other eukaryotic microorganisms. This unusual growth phenotype was due to an incomplete glucose repression of the function of the glyoxylate cycle, as shown by the lack of growth in that medium of double pyc1 icl1 mutants lacking both pyruvate carboxylase and isocitrate lyase activity. These mutants grew when glutamate, aspartate, or Casamino Acids were added to the glucose-ammonium medium. The cDNA from the Y. lipolytica PYC1 gene complemented the growth defect of a Saccharomyces cerevisiae pyc1 pyc2 mutant, but introduction of either the S. cerevisiae PYC1 or PYC2 gene into Y. lipolytica did not result in detectable pyruvate carboxylase activity or in growth on glucose-ammonium of a Y. lipolytica pyc1 icl1 double mutant. PMID:15701798

Flores, Carmen-Lisset; Gancedo, Carlos



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.



Chemical synthesis of 20S-hydroxyvitamin D3, which shows anti-proliferative activity  

PubMed Central

20S-hydroxyvitamin D3 (20S-(OH)D3), an in vitro product of vitamin D3 metabolism by the cytochrome P450scc, was recently isolated, identified and shown to possess antiproliferative activity without inducing hypercalcemia. The enzymatic production of 20S-(OH)D3 is tedious, expensive, and cannot meet the requirements for extensive chemical and biological studies. Here we report for the first time the chemical synthesis of 20S-(OH)D3 which exhibited biological properties characteristic of the P450scc-generated compound. Specifically, it was hydroxylated to 20,23-dihydroxyvitamin D3 and 17,20-dihydroxyvitamin D3 by P450scc and was converted to 1?,20-dihydroxyvitamin D3 by CYP27B1. It inhibited proliferation of human epidermal keratinocytes with lower potency than 1?,25-dihydroxyvitamin D3 (1,25(OH)2D3) in normal epidermal human keratinocytes, but with equal potency in immortalized HaCaT keratinocytes. It also stimulated VDR gene expression with similar potency to 1,25(OH)2D3, and stimulated involucrin (a marker of differentiation) and CYP24 gene expression, showing a lower potency for the latter gene than 1,25(OH)2D3. Testing performed with hamster melanoma cells demonstrated a dose-dependent inhibition of cell proliferation and colony forming capabilities similar or more pronounced than those of 1,25(OH)2D3. Thus, we have developed a chemical method for the synthesis of 20S-(OH)D3, which will allow the preparation of a series of 20S-(OH)D3 analogs to study structure-activity relationships to further optimize this class of compound for therapeutic use.

Li, Wei; Chen, Jianjun; Janjetovic, Zorica; Kim, Tae-Kang; Sweatman, Trevor; Lu, Yan; Zjawiony, Jordan; Tuckey, Robert C; Miller, Duane; Slominski, Andrzej



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 Ĺ). 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 ( or via

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.



Simultaneous detection of different Rhizobium strains marked with either the Escherichia coli gusA gene or the Pyrococcus furiosus celB gene.  

PubMed Central

A new marker system for gram-negative bacteria was developed on the basis of the celB gene from the hyperthermophilic archaeon Pyrococcus furiosus, which encodes a thermostable beta-glucosidase with a high level of beta-galactosidase activity. The celB gene is highly suitable as a marker for studying plant-bacterium interaction because endogenous background beta-glucosidase and beta-galactosidase enzyme activity can readily be inactivated by heat and because inexpensive substrates for detection are commercially available. Two celB-expressing transposons were constructed for use in ecological studies of a variety of gram-negative bacteria. The combined use of the gusA marker gene and celB allowed the simultaneous detection of several Rhizobium strains on a plant, and multiple-strain occupancy of individual modules also could be easily detected.

Sessitsch, A; Wilson, K J; Akkermans, A D; de Vos, W M



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



Apigenin shows synergistic anticancer activity with curcumin by binding at different sites of tubulin.  


Apigenin, a natural flavone, present in many plants sources, induced apoptosis and cell death in lung epithelium cancer (A549) cells with an IC50 value of 93.7 ± 3.7 ?M for 48 h treatment. Target identification investigations using A549 cells and also in cell-free system demonstrated that apigenin depolymerized microtubules and inhibited reassembly of cold depolymerized microtubules of A549 cells. Again apigenin inhibited polymerization of purified tubulin with an IC50 value of 79.8 ± 2.4 ?M. It bounds to tubulin in cell-free system and quenched the intrinsic fluorescence of tubulin in a concentration- and time-dependent manner. The interaction was temperature-dependent and kinetics of binding was biphasic in nature with binding rate constants of 11.5 × 10(-7) M(-1) s(-1) and 4.0 × 10(-9) M(-1) s(-1) for fast and slow phases at 37 °C, respectively. The stoichiometry of tubulin-apigenin binding was 1:1 and binding the binding constant (Kd) was 6.08 ± 0.096 ?M. Interestingly, apigenin showed synergistic anti-cancer effect with another natural anti-tubulin agent curcumin. Apigenin and curcumin synergistically induced cell death and apoptosis and also blocked cell cycle progression at G2/M phase of A549 cells. The synergistic activity of apigenin and curcumin was also apparent from their strong depolymerizing effects on interphase microtubules and inhibitory effect of reassembly of cold depolymerized microtubules when used in combinations, indicating that these ligands bind to tubulin at different sites. In silico modeling suggested apigenin bounds at the interphase of ?-?-subunit of tubulin. The binding site is 19 Ĺ in distance from the previously predicted curcumin binding site. Binding studies with purified protein also showed both apigenin and curcumin can simultaneously bind to purified tubulin. Understanding the mechanism of synergistic effect of apigenin and curcumin could be helped to develop anti-cancer combination drugs from cheap and readily available nutraceuticals. PMID:23485682

Choudhury, Diptiman; Ganguli, Arnab; Dastidar, Debabrata Ghosh; Acharya, Bipul R; Das, Amlan; Chakrabarti, Gopal



Mice lacking brain-type creatine kinase activity show defective thermoregulation  

PubMed Central

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°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.

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



Regional Brain Activation during Meditation Shows Time and Practice Effects: An Exploratory FMRI Study  

Microsoft Academic Search

Meditation involves attentional regulation and may lead to increased activity in brain regions associated with attention such as dorsal lateral prefrontal cortex (DLPFC) and anterior cingulate cortex (ACC). Using functional magnetic resonance imaging, we examined whether DLPFC and ACC were activated during meditation. Subjects who meditate were recruited and scanned on a 3.0 Tesla scanner. Subjects meditated for four sessions

E. Baron Short; Samet Kose; Qiwen Mu; Jeffery Borckardt; Andrew Newberg; Mark S. George; F. Andrew Kozel



Regional Brain Activation During Meditation Shows Time and Practice Effects: An Exploratory FMRI Studyy  

Microsoft Academic Search

Meditation involves attentional regulation and may lead to increased activity in brain regions associated with attention such as dorsal lateral prefrontal cortex (DLPFC) and anterior cingulate cortex (ACC). Using functional magnetic resonance imaging, we examined whether DLPFC and ACC were activated during meditation. Subjects who meditate were recruited and scanned on a 3.0 Tesla scanner. Subjects meditated for four sessions

E. Baron Short; Samet Kose; Qiwen Mu; Jeffery Borckardt; Andrew Newberg; Mark S. George; F. Andrew Kozel


RNA sequencing shows no dosage compensation of the active X-chromosome  

Microsoft Academic Search

Mammalian cells from both sexes typically contain one active X chromosome but two sets of autosomes. It has previously been hypothesized that X-linked genes are expressed at twice the level of autosomal genes per active allele to balance the gene dose between the X chromosome and autosomes (termed 'Ohno's hypothesis'). This hypothesis was supported by the observation that microarray-based gene

Yuanyan Xiong; Xiaoshu Chen; Zhidong Chen; Xunzhang Wang; Suhua Shi; Xueqin Wang; Jianzhi Zhang; Xionglei He



Long-term global hurricane activity record shows periodic highs and lows  

NASA Astrophysics Data System (ADS)

In the long-record of hurricane activity, which stretches back roughly 7500 years, the frequency of large tropical cyclones has varied, with distinct periods of elevated or depressed activity lasting hundreds to thousands of years. Of the 12 sites that provide the basis for this record, 6 measure the activity in the tropical North Atlantic, 1 serves as a history of the northwestern Pacific, and 5 sites around Australia represent the southwestern Pacific Ocean and the Indian Ocean. In previous research, scientists used the individual records from northern hemispheric sites to investigate the connection between long-term tropical cyclone activity and climate systems such as the El Nińo Southern Oscillation (ENSO). Nott and Forsyth, however, used the records from all 12 sites to ascertain a measure of long-term global hurricane activity.

Schultz, Colin





We extended our previous exploration of sulfur bridges as bioisosteric replacements for atoms forming the bridge between the aromatic rings of combretastatin A-4. Employing coupling reactions between 5-iodo-1,2,3-trimethoxybenzene and substituted thiols, followed by oxidation to sulfones with m-CPBA, different locations for attaching the sulfur atom to ring A through the synthesis of nine compounds were examined. Antitubulin activity was performed with electrophoretically homogenous bovine brain tubulin, and activity occurred with the 1,2,3-trimethoxy-4-[(4-methoxyphenyl)thio]benzene (12), while the other compounds were inactive. The compounds were also tested for leishmanicidal activity using promastigote forms of Leishmania braziliensis (MHOM/BR175/M2904), and the greatest activity was observed with 1,2,3-trimethoxy-4-(phenylthio)benzene (10) and 1,2,3-trimethoxy-4-[(4-methoxyphenyl) sulfinyl]benzene (15). PMID:23766547

Dos Santos, Edson Dos A; Prado, Paulo C; de Carvalho, Wanderley R; de Lima, Ricardo V; Beatriz E, Adilson; de Lima, Dęnis P; Hamel, Ernest; Dyba, Marzena A; Albuquerque, Sergio



Interconnected Pt-Nanodendrite/DNA/Reduced-Graphene-Oxide Hybrid Showing Remarkable Oxygen Reduction Activity and Stability.  


Controlling the morphology and size of platinum nanodendrites (PtDs) is a key factor in improving their catalytic activity and stability. Here, we report the synthesis of PtDs on genomic-double-stranded-DNA/reduced-graphene-oxide (gdsDNA/rGO) by the NaBH4 reduction of H2PtCl6 in the presence of plant gdsDNA. Compared to industrially adopted catalysts (i.e., state-of-the-art Pt/C catalyst, Pt/rGO, Pt3Co, etc.), the as-synthesized PtDs/gdsDNA/rGO hybrid displays very high oxygen reduction reaction (ORR) catalytic activities (much higher than the 2015 U.S. Department of Energy (DOE) target values), which are the rate-determining steps in electrochemical energy devices, in terms of onset-potential, half-wave potential, specific-activity, mass-activity, stability, and durability. Moreover, the hybrid exhibits a highly stable mass activity for the ORR over a wide pH range of 1-13. These exceptional properties would make the hybrid applicable in next-generation electrochemical energy devices. PMID:23991709

Tiwari, Jitendra N; Kemp, Kingsley Christian; Nath, Krishna; Tiwari, Rajanish N; Nam, Hong-Gil; Kim, Kwang S



Isolation of novel Bacillus species showing high mosquitocidal activity against several mosquito species.  


Two novel mosquitocidal bacteria, VB17 and VB24, identified as new Bacillus species were isolated from dead mosquito larvae obtained in Florida aquatic habitats. Gas chromatographic analysis of fatty acid methyl esters (GC-FAME) and 16S rRNA sequencing indicated that VB24 is closely related to Bacillus sphaericus whereas VB17 does not have a close relationship with either Bacillus thuringiensis or B. sphaericus. Both isolates were significantly more active than B. sphaericus 2362 against Aedes taeniorhynchus, Anopheles quadrimaculatus, Culex quinquefasciatus larvae, and as active as B. sphaericus 2362 against Anopheles gambiae. Interestingly, however, both were not active against Aedes aegypti larvae, indicating some level of insecticidal specificity. PMID:21276795

Hayes, Sabrina R; Hudon, Michael; Park, Hyun-Woo



A transgenic apple callus showing reduced polyphenol oxidase activity and lower browning potential.  


Polyphenol oxidase (PPO) is responsible for enzymatic browning of apples. Apples lacking PPO activity might be useful not only for the food industry but also for studies of the metabolism of polyphenols and the function of PPO. Transgenic apple calli were prepared by using Agrobacterium tumefaciens carrying the kanamycin (KM) resistant gene and antisense PPO gene. Four KM-resistant callus lines were obtained from 356 leaf explants. Among these transgenic calli, three calli grew on the medium containing KM at the same rate as non-transgenic callus on the medium without KM. One callus line had an antisense PPO gene, in which the amount and activity of PPO were reduced to half the amount and activity in non-transgenic callus. The browning potential of this line, which was estimated by adding chlorogenic acid, was also half the browning potential of non-transgenic callus. PMID:11302173

Murata, M; Nishimura, M; Murai, N; Haruta, M; Homma, S; Itoh, Y



Screening of Exiguobacterium acetylicum from soil samples showing enantioselective and alkalotolerant esterase activity  

Microsoft Academic Search

About 3,000 bacterial colonies with esterase activities were isolated from soil samples by enrichment culture and halo-size\\u000a on Luria broth-tributyrin (LT) plates. The colonies were assayed for esterase activity in microtiter plates using enantiomerically\\u000a pure (R)- and (S)-2-phenylbutyric acid resorufin ester (2PB-O-res) as substrates. Two enantioselective strains (JH2 and JH13) were selected\\u000a by the ratio of initial rate of hydrolysis

Bum-Yeol Hwang; Ji-Hyun Kim; Juhan Kim; Byung-Gee Kim



Cysteamine, the natural metabolite of pantetheinase, shows specific activity against Plasmodium  

Microsoft Academic Search

In mice, loss of pantetheinase activity causes susceptibility to infection with Plasmodium chabaudi AS. Treatment of mice with the pantetheinase metabolite cysteamine reduces blood-stage replication of P. chabaudi and significantly increases survival. Similarly, a short exposure of Plasmodium to cysteamine ex vivo is sufficient to suppress parasite infectivity in vivo. This effect of cysteamine is specific and not observed with

Gundula Min-Oo; Kodjo Ayi; Silayuv E. Bongfen; Mifong Tam; Irena Radovanovic; Susan Gauthier; Helton Santiago; Antonio Gigliotti Rothfuchs; Ester Roffę; Alan Sher; Alaka Mullick; Anny Fortin; Mary M. Stevenson; Kevin C. Kain; Philippe Gros



Antibodies to Meningococcal H.8 (Lip) Antigen Fail to Show Bactericidal Activity.  

National Technical Information Service (NTIS)

Purified H.8 (Lip) antigen was coupled to tresyl-activated Sepharose 4B and used in affinity columns to purify anti-Lip antibodies from convalescent patient sera and from immune rabbit sera. Affinity-purified anti-Lip antibodies isolated from two convales...

A. K. Bhattacharjee E. E. Moran W. D. Zollinger



Patients treated with high-dose intravenous immunoglobulin show selective activation of regulatory T cells.  


Intravenous immunoglobulin (IVIg) is used to treat autoimmune and systemic inflammatory diseases caused by derailment of humoral and cellular immunity. In this study we investigated whether IVIg treatment can modulate regulatory T cells (Tregs ) in humans in vivo. Blood was collected from IVIg-treated patients with immunodeficiency or autoimmune disease who were treated with low-dose (n = 12) or high-dose (n = 15) IVIg before, immediately after and at 7 days after treatment. Percentages and activation status of circulating CD4(+) CD25(+) forkhead box protein 3 (FoxP3(+)) Tregs and of conventional CD4(+) FoxP3(-) T-helper cells (Tconv) were measured. The suppressive capacity of Tregs purified from blood collected at the time-points indicated was determined in an ex-vivo assay. High-dose, but not low-dose, IVIg treatment enhanced the activation status of circulating Tregs , as shown by increased FoxP3 and human leucocyte antigen D-related (HLA-DR) expression, while numbers of circulating Tregs remained unchanged. The enhanced activation was sustained for at least 7 days after infusion, and the suppressive capacity of purified Tregs was increased from 41 to 70% at day 7 after IVIg treatment. The activation status of Tconv was not affected by IVIg. We conclude that high-dose IVIg treatment activates Tregs selectively and enhances their suppressive function in humans in vivo. This effect may be one of the mechanisms by which IVIg restores imbalanced immune homeostasis in patients with autoimmune and systemic inflammatory disorders. PMID:23607448

Tjon, A S W; Tha-In, T; Metselaar, H J; van Gent, R; van der Laan, L J W; Groothuismink, Z M A; te Boekhorst, P A W; van Hagen, P M; Kwekkeboom, J



Declassified American Government Documents Show a Broad and In-Depth Interest in Soviet Space Activities  

NASA Astrophysics Data System (ADS)

Back in 1993, when this author was able to acquire one of the first US National Intelligence Estimate (NIE) documents declassified on the Soviet Space Program [1], there was a dearth of materials concerning the USSR's space activities from a US intelligence perspective. Now, a decade on, the situation has dramatically changed. As a plethora of US government agencies labor to disgorge their materials from the era of the Cold War, space historians and observers now have access to many new documents, which shed both exciting and new light on Soviet space activities, and how the US viewed as well as interpreted them. Coupled with the fact that many of these are now available to be read via the Internet, and with most of the documents available only recently, a new era in space history research is now in hand. This article is intended to provide a broad overview of what is now available, and mention some highlights.

Pesavento, P.


Basic N-interlinked imipramines show apoptotic activity against malignant cells including Burkitt's lymphoma.  


We here report the synthesis of ethylene glycol N-interlinked imipramine dimers of various lengths from the tricyclic antidepressant desipramine via an amide coupling reaction followed by reduction with lithium aluminium hydride. The target molecules were found to be potent inhibitors of cellular viability while inducing cell type specific death mechanisms in three cancer cell lines including a highly chemoresistant Burkitt's lymphoma cell line. Basic amine analogues were found to be important for increased potency. Imipramine and desipramine were also tested for apoptotic activity and were found to be much less active than the novel dimeric compounds. Imipramine dimers were only found to be moderate inhibitors of the human serotonin transporter (hSERT) having IC(50) values in the micromolar region whilst the induction of cell death occurred independently of hSERT expression. These results demonstrate the potential of newly designed and synthesised imipramines derivatives for use against malignant cells, including those resistant to standard chemotherapy. PMID:23385211

Bright, Sandra A; Brinkř, Anne; Larsen, Maja Thim; Sinning, Steffen; Williams, D Clive; Jensen, Henrik H



Long-period accelerometer monitoring shows the role of physical activity in overweight and obesity  

Microsoft Academic Search

CONTEXT:Physical activity (PA) plays an important role in obesity. A new accelerometer has been developed to assess total energy expenditure as well as PA.OBJECTIVE:To investigate the association of PA with overweight and obesity in Japanese men and women, a large cross-sectional study was performed using a single-axis accelerometer.DESIGN, SETTING AND PARTICIPANTS:Population-based cross-sectional study of Japanese 18–84 y of age. Height,

M Yoshioka; M Ayabe; T Yahiro; H Higuchi; Y Higaki; J St-Amand; H Miyazaki; Y Yoshitake; M Shindo; H Tanaka



Keratinolytic proteases of Bacillus species isolated from the Amazon basin showing remarkable de-hairing activity  

Microsoft Academic Search

Three keratinolytic Bacillus spp. isolated from the Brazilian Amazon basin were characterized. The strains P6, P7 and P11 were identified based on morphological\\u000a and biochemical characteristics and 16S rDNA sequences. P6, P7 and P11 sequences shared more than 99% similarity with B. subtilis, B. amyloliquefaciens and B. velesensis. The keratinases produced by these bacteria were active on azokeratin and degradation

Janice Luhering Giongo; Françoise S. Lucas; Franciani Casarin; Philipp Heeb; Adriano Brandelli



Sulfide : quinone oxidoreductase (SQR) from the lugworm Arenicola marina shows cyanide- and thioredoxin-dependent activity.  


The lugworm Arenicola marina inhabits marine sediments in which sulfide concentrations can reach up to 2 mM. Although sulfide is a potent toxin for humans and most animals, because it inhibits mitochondrial cytochrome c oxidase at micromolar concentrations, A. marina can use electrons from sulfide for mitochondrial ATP production. In bacteria, electron transfer from sulfide to quinone is catalyzed by the membrane-bound flavoprotein sulfide : quinone oxidoreductase (SQR). A cDNA from A. marina was isolated and expressed in Saccharomyces cerevisiae, which lacks endogenous SQR. The heterologous enzyme was active in mitochondrial membranes. After affinity purification, Arenicola SQR isolated from yeast mitochondria reduced decyl-ubiquinone (K(m) = 6.4 microm) after the addition of sulfide (K(m) = 23 microm) only in the presence of cyanide (K(m) = 2.6 mM). The end product of the reaction was thiocyanate. When cyanide was substituted by Escherichia coli thioredoxin and sulfite, SQR exhibited one-tenth of the cyanide-dependent activity. Six amino acids known to be essential for bacterial SQR were exchanged by site-directed mutagenesis. None of the mutant enzymes was active after expression in yeast, implicating these amino acids in the catalytic mechanism of the eukaryotic enzyme. PMID:18248458

Theissen, Ursula; Martin, William



Cyclic GMP-activated channels of rod photoreceptors show neither fast nor slow desensitization.  


Desensitization of cGMP-activated channels was examined in excised, inside-out patches obtained from rod photoreceptors. Cyclic GMP was applied using a rapid-flow system in which concentration jumps are complete within 10-50 ms. In outer-segment patches containing many channels, the cGMP-dependent conductance reached a steady plateau that was maintained for tens of seconds in the presence of cGMP; thus, there was no indication of slow desensitization. However, rapid desensitization on the scale of milliseconds could not be ruled out because of limited speed of access of cGMP to the inner face of the patch membrane. To test for rapid desensitization, inner-segment patches containing only a single cGMP-activated channel were used. In these one-channel patches, there was no change in activity of the channel with time from its earliest onset after application of cGMP, indicating that rapid desensitization also did not occur. PMID:1702987

Watanabe, S; Matthews, G



NF-?B dynamics show digital activation and analog information processing in cells  

NASA Astrophysics Data System (ADS)

Cells operate in ever changing environments using extraordinary communication capabilities. Cell-to-cell communication is mediated by signaling molecules that form spatiotemporal concentration gradients, which requires cells to respond to a wide range of signal intensities. We used high-throughput microfluidic cell culture, quantitative gene expression analysis and mathematical modeling to investigate how single mammalian cells respond to different concentrations of the signaling molecule TNF-? via the transcription factor NF-?B. We measured NF-?B activity in thousands of live cells under TNF-? doses covering four orders of magnitude. In contrast to population studies, the activation is a stochastic, switch-like process at the single cell level with fewer cells responding at lower doses. The activated cells respond fully and express early genes independent of the TNF-? concentration, while only high dose stimulation results in the expression of late genes. Cells also encode a set of analog parameters such as the NF-?B peak intensity, response time and number of oscillations to modulate the outcome. We developed a stochastic model that reproduces both the digital and analog dynamics as well as the gene expression profiles at all measured conditions, constituting a broadly applicable model for TNF-? induced NF-?B signaling in various types of cells.

Tay, Savas; Hughey, Jake; Lee, Timothy; Lipniacki, Tomasz; Covert, Markus; Quake, Stephen



Phosphorylated smooth muscle heavy meromyosin shows an open conformation linked to activation  

PubMed Central

Smooth muscle myosin and heavy meromyosin (smHMM) are activated by regulatory light chain (RLC) phosphorylation but the mechanism remains unclear. Dephosphorylated, inactive smHMM assumes a closed conformation with asymmetric intramolecular head-head interactions between motor domains. The “free head” can bind to actin, but the actin-binding interface of the “blocked head” is involved in interactions with the free head. We report here a 3-D structure for phosphorylated, active smHMM obtained using electron crystallography of 2-D arrays. Head-head interactions of phosphorylated smHMM resemble those found in the dephosphorylated state, but occur between different molecules, not within the same molecule. The light chain binding domain structure of phosphorylated smHMM differs markedly from that of the “blocked” head of dephosphorylated smHMM. We hypothesize that RLC phosphorylation opens the inhibited conformation primarily by its effect on the blocked head. Singly phosphorylated smHMM is not compatible with the closed conformation if the blocked head is phosphorylated. This concept has implications for the extent of myosin activation at low levels of phosphorylation in smooth muscle.

Baumann, Bruce A. J.; Taylor, Dianne W.; Huang, Zhong; Tama, Florence; Fagnant, Patricia M.; Trybus, Kathleen M.; Taylor, Kenneth A.



Zinc oxide nanoparticles show antidiabetic activity in streptozotocin-induced Types-1 and 2 diabetic rats.  


Aim: The correlation of diabetes and an imbalance in zinc homeostasis makes zinc-based therapy an attractive proposition. In this study, zinc oxide nanoparticles were evaluated for antidiabetic effects and safety. Materials & methods: Zinc oxide nanoparticles (1, 3 and 10 mg/kg) were tested for antidiabetic activity in streptozotocin-induced Types-1 and 2 diabetic rats. Single-dose pharmacokinetic study, cytotoxicity, hemolysis, acute and subacute toxicity tests, and mechanism-of-action studies were performed. Results: Oral administration of zinc oxide nanoparticles resulted in significant antidiabetic effects - that is, improved glucose tolerance, higher serum insulin (70%), reduced blood glucose (29%), reduced nonesterified fatty acids (40%) and reduced triglycerides (48%). Nanoparticles were systemically absorbed resulting in elevated zinc levels in the liver, adipose tissue and pancreas. Increased insulin secretion and superoxide dismutase activity were also seen in rat insulinoma (RIN5f) cells. Nanoparticles were safe up to a 300 mg/kg dose in rats. Conclusion: Zinc oxide nanoparticles are a promising antidiabetic agent warranting further studies. Original submitted 9 July 2012; Revised submitted 27 November 2012. PMID:23427863

Umrani, Rinku D; Paknikar, Kishore M



X-ray microtomography shows pore structure and tortuosity in alkali-activated binders  

SciTech Connect

Durability of alkali-activated binders is of vital importance in their commercial application, and depends strongly on microstructure and pore network characteristics. X-ray microtomography ({mu}CT) offers, for the first time, direct insight into microstructural and pore structure characteristics in three dimensions. Here, {mu}CT is performed on a set of sodium metasilicate-activated fly ash/slag blends, using a synchrotron beamline instrument. Segmentation of the samples into pore and solid regions is then conducted, and pore tortuosity is calculated by a random walker method. Segmented porosity and diffusion tortuosity are correlated, and vary as a function of slag content (slag addition reduces porosity and increases tortuosity), and sample age (extended curing gives lower porosity and higher tortuosity). This is particularly notable for samples with {>=} 50% slag content, where a space-filling calcium (alumino)silicate hydrate gel provides porosity reductions which are not observed for the sodium aluminosilicate ('geopolymer') gels which do not chemically bind water of hydration.

Provis, John L., E-mail: [Department of Chemical and Biomolecular Engineering, University of Melbourne, Victoria 3010 (Australia); Myers, Rupert J.; White, Claire E. [Department of Chemical and Biomolecular Engineering, University of Melbourne, Victoria 3010 (Australia); Rose, Volker [Advanced Photon Source, Argonne National Laboratory, 9700 South Cass Ave, Argonne IL 60439 (United States); Deventer, Jannie S.J. van [Department of Chemical and Biomolecular Engineering, University of Melbourne, Victoria 3010 (Australia); Zeobond Pty Ltd, P.O. Box 210, Somerton, Victoria 3062 (Australia)



Variation in GUS activity in vegetatively propagated Hevea brasiliensis transgenic plants  

Microsoft Academic Search

Hevea brasiliensis transgenic plants are regenerated from transgenic callus lines by somatic embryogenesis. Somatic embryogenesis is not yet\\u000a available for commercial propagation of Hevea clones, which requires conventional grafting of buds on rootstock seedlings (budding). The stability of transgene expression\\u000a in budded plants is therefore necessary for further development of genetic engineering in rubber trees. Transgene expression\\u000a was assessed by

Ludovic Lardet; Julie Leclercq; Elise Bénistan; Florence Dessailly; Gérald Oliver; Florence Martin; Pascal Montoro


Extracellular matrix metalloproteinase inducer shows active perivascular cuffs in multiple sclerosis.  


Inflammatory perivascular cuffs are comprised of leucocytes that accumulate in the perivascular space around post-capillary venules before their infiltration into the parenchyma of the central nervous system. Inflammatory perivascular cuffs are commonly found in the central nervous system of patients with multiple sclerosis and in the animal model experimental autoimmune encephalomyelitis. Leucocytes that accumulate in the perivascular space secrete matrix metalloproteinases that aid their transmigration into the neural parenchyma. We described previously that the upstream inducer of matrix metalloproteinase expression, extracellular matrix metalloproteinase inducer (CD147), was elevated in experimental autoimmune encephalomyelitis, and that its inhibition reduced leucocyte entry into the central nervous system. Here we investigated whether the expression of extracellular matrix metalloproteinase inducer varies with the temporal evolution of lesions in murine experimental autoimmune encephalomyelitis, whether it was uniformly upregulated across multiple sclerosis specimens, and whether it was a feature of inflammatory perivascular cuffs in multiple sclerosis lesions. In experimental autoimmune encephalomyelitis, elevation of extracellular matrix metalloproteinase inducer was correlated with the appearance and persistence of clinical signs of disease. In both murine and human samples, extracellular matrix metalloproteinase inducer was detected on endothelium in healthy and disease states but was dramatically increased in and around inflammatory perivascular cuffs on leucocytes, associated with matrix metalloproteinase expression, and on resident cells including microglia. Leucocyte populations that express extracellular matrix metalloproteinase inducer in multiple sclerosis lesions included CD4+ and CD8+ T lymphocytes, B lymphocytes and monocyte/macrophages. The extra-endothelial expression of extracellular matrix metalloproteinase inducer was a marker of the activity of lesions in multiple sclerosis, being present on leucocyte-containing perivascular cuffs but not in inactive lesions. By using a function-blocking antibody, we implicate extracellular matrix metalloproteinase inducer in the adhesion of leucocytes to endothelial cells and determined that its activity was more crucial on leucocytes than on endothelium in leucocyte-endothelial cell engagement in vitro. Extracellular matrix metalloproteinase inducer activity regulated the level of alpha 4 integrin on leucocytes through a mechanism associated with nuclear factor ?B signalling. Blocking extracellular matrix metalloproteinase inducer attenuated the transmigration of monocytes and B lymphocytes across a model of the blood-brain barrier in culture. In summary, we describe the prominence of extracellular matrix metalloproteinase inducer in central nervous system inflammatory perivascular cuffs, emphasize its dual role in matrix metalloproteinase induction and leucocyte adhesion, and highlight the elevation of extracellular matrix metalloproteinase inducer as an orchestrator of the infiltration of leucocytes into the central nervous system parenchyma. PMID:23687119

Agrawal, Smriti M; Williamson, Jacqueline; Sharma, Ritu; Kebir, Hania; Patel, Kamala; Prat, Alexandre; Yong, V Wee



The structure of two IgMs showing different activity from a patient with Waldenstrom's macroglobulinaemia.  

PubMed Central

The two monoclonal IgMs (IgM1 and IgM2) were characterized from a patient Waldenström's macroglobulinaemia that resulted in a gammapathy. Heavy and light chains were isolated from the IgM. The complete primary structure of the two light chains and the NH2-terminal region of the two heavy chain molecules were determined. The sequence data indicated that the heavy and light chains from both IgMs belong to the same (III and II) lambda subgroups. By testing their antibody activity it was found by immunofluorescence and immunoblotting that only IgM2 reacts with an intermediate filament protein. Images Fig. 1 Fig. 4 Fig. 5 Fig. 6 Fig. 7

Mendez, E; Osuna, C; Sanchez, A; Revilla, Y; Soriano, F; Montalban, C; Segui, J; Avila, J



? sulphate PNA (PNA S): Highly Selective DNA Binding Molecule Showing Promising Antigene Activity  

PubMed Central

Peptide Nucleic Acids (PNAs), nucleic acid analogues showing high stability to enzyme degradation and strong affinity and specificity of binding toward DNA and RNA are widely investigated as tools to interfere in gene expression. Several studies have been focused on PNA analogues with modifications on the backbone and bases in the attempt to overcome solubility, uptake and aggregation issues. ? PNAs, PNA derivatives having a substituent in the ? position of the backbone show interesting properties in terms of secondary structure and affinity of binding toward complementary nucleic acids. In this paper we illustrate our results obtained on new analogues, bearing a sulphate in the ? position of the backbone, developed to be more DNA-like in terms of polarity and charge. The synthesis of monomers and oligomers is described. NMR studies on the conformational properties of monomers and studies on the secondary structure of single strands and triplexes are reported. Furthermore the hybrid stability and the effect of mismatches on the stability have also been investigated. Finally, the ability of the new analogue to work as antigene, interfering with the transcription of the ErbB2 gene on a human cell line overexpressing ErbB2 (SKBR3), assessed by FACS and qPCR, is described.

Avitabile, Concetta; Moggio, Loredana; Malgieri, Gaetano; Capasso, Domenica; Di Gaetano, Sonia; Saviano, Michele; Pedone, Carlo; Romanelli, Alessandra



Biolistic-mediated DNA delivery and transient expression of GUS in hypocotyls of Feronia limonia L.—A fruit tree  

Microsoft Academic Search

Feronia limonia L. (Rutaceae) is an important fruit tree of arid horticulture. To be able to introduce genes of interest for its improvement, biolistic-mediated DNA delivery method has been standardized. The hypocotyls explants were bombarded with plasmid pBI121 having gus reporter gene driven by CaMV 35S promoter and npt II under control of nos promoter as selectable marker. The best

S D Purohit; S Raghuvanshi; A K Tyagi


Specific Isoforms of Translation Initiation Factor 4GI Show Differences in Translational Activity? †  

PubMed Central

The eukaryotic initiation factor (eIF) 4GI gene locus (eIF4GI) contains three identified promoters, generating alternately spliced mRNAs, yielding a total of five eIF4GI protein isoforms. Although eIF4GI plays a critical role in mRNA recruitment to the ribosomes, little is known about the functions of the different isoforms, their partner binding capacities, or the role of the homolog, eIF4GII, in translation initiation. To directly address this, we have used short interfering RNAs (siRNAs) expressed from DNA vectors to silence the expression of eIF4GI in HeLa cells. Here we show that reduced levels of specific mRNA and eIF4GI isoforms in HeLa cells promoted aberrant morphology and a partial inhibition of translation. The latter reflected dephosphorylation of 4E-BP1 and decreased eIF4F complex levels, with no change in eIF2? phosphorylation. Expression of siRNA-resistant Myc-tagged eIF4GI isoforms has allowed us to show that the different isoforms exhibit significant differences in their ability to restore translation rates. Here we quantify the efficiency of eIF4GI promoter usage in mammalian cells and demonstrate that even though the longest isoform of eIF4GI (eIF4GIf) was relatively poorly expressed when reintroduced, it was more efficient at promoting the translation of cellular mRNAs than the more highly expressed shorter isoforms used in previous functional studies.

Coldwell, Mark J.; Morley, Simon J.



Dammarenolic acid, a secodammarane triterpenoid from Aglaia sp. shows potent anti-retroviral activity in vitro.  


Screening of a panel of purified compounds isolated from Aglaia sp. (Meliaceae) for inhibition of early steps in the lentiviral replication cycle led to the identification of the 3, 4-secodammarane triterpenoid, ignT1, which inhibited HIV-1 infection potently (IC(50)=0.48microg/ml), while cytotoxic effects and inhibition of cell proliferation were only observed at concentrations exceeding 10.69microg/ml. Time of addition experiments revealed similar kinetics to the non-nucleoside RT-inhibitor (NNRTI), Nevirapine, although the latter was significantly less cytotoxic. However, unlike Nevirapine, dammarenolic acid also potently inhibited the in vitro replication of other retroviruses, including Simian immunodeficiency virus and Murine leukemic virus in vector-based antiviral screening studies. Interestingly, the methyl ester analogue of dammarenolic acid-methyldammarenolate had no anti-HIV-1 activity. Cell cycle analysis revealed that ignT1 arrests HeLa cells at the S and G2/M phase. These results strongly suggest that dammarenolic acid could be a promising lead compound for the development of novel anti-retrovirals. PMID:19962871

Esimone, Charles O; Eck, Gero; Nworu, Chukwuemeka S; Hoffmann, Dennis; Uberla, Klaus; Proksch, Peter



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



Recovery of phenotypically normal transgenic plants of Brassica oleracea upon Agrobacterium rhizogenes-mediated co-transformation and selection of transformed hairy roots by GUS assay  

Microsoft Academic Search

In this paper we describe the production of transgenic broccoli and cauliflower with normal phenotype using an Agrobacterium rhizogenes-mediated transformation system with efficient selection for transgenic hairy-roots. Hypocotyls were inoculated with Agrobacterium strain A4T harbouring the bacterial plasmid pRiA4 and a binary vector pMaspro::GUS whose T-DNA region carried the gus reporter gene. pRiA4 transfers TL sequences carrying the rol genes

I. J. Puddephat; H. T. Robinson; T. M. Fenning; D. J. Barbara; A. Morton; D. A. C. Pink



Star Show Classroom Activity  

NSDL National Science Digital Library

In this lesson from Math Machines, the class will "plan and conduct a simulated astronomical observing session to photgraph a variety of star types." The instructor will set up several "stars" around the classroom, and students will then set up a telescope location and estimate the altitude and azimuth to photograph each star. A participant handout (including worksheet) and facilitator notes are made available for download in DOC file format. Links to calculator programs are also included.

Thomas, Fred



Evidence from northwest European bogs shows ‘Little Ice Age’ climatic changes driven by variations in solar activity  

Microsoft Academic Search

Fluctuations in Holocene atmospheric radiocarbon concentrations have been shown to be due to variations in solar activity. Analyses of both 10Be and 14C nuclides con” rm that production-rate changes during the Holocene were largely modulated by solar activity. Analyses of peat samples from two intact European ombrotrophic bogs show that climatic deteriorations during the ‘Little Ice Age’ are associated with

Dmitri Mauquoy; Bas van Geel; Maarten Blaauw; Johannes van der Plicht



TV as a Multimedia Synchronous Communication for Cooking and Eating Activities: Analysis of TV Cooking Shows in Hong Kong  

Microsoft Academic Search

TV can be considered to be a synchronous tool for multimedia communication to deliver contents like cooking and eating activities. In this paper, we propose a framework of multimedia for cooking and eating activities. It consists of three facets namely contents, multimedia elements and communication. We demonstrate the framework with an example. Under this framework, we analyzed TV cooking shows

T. W. Lai-Yeung; S. W. W. So



Monocytes Constitute the Only Peripheral Blood Cell Population Showing an Increased Burst Activity in Multiple Sclerosis Patients  

Microsoft Academic Search

We examined peripheral mononuclear cells (PMNC), peripheral polymorphonuclear cells (PPNC), adherent mononuclear cells, non-adherent mononuclear cells, non-phagocytic mononuclear cells and non-phagocytic Percoll-fractionated mononuclear cells of multiple sclerosis (MS) patients for their spontaneous burst activity (B A). According to previous results PMNC of MS patients showed a markedly increased BA (p = 0.0002); PPNC, however, did not show significantly elevated values

Klaus P. Hammann; Hanns C. Hopf



Cistaceae aqueous extracts containing ellagitannins show antioxidant and antimicrobial capacity, and cytotoxic activity against human cancer cells.  


Roots and aerial parts of Cistaceae have been used since ancient times in the Mediterranean cultures for its medicinal properties. In this study, phenolic and tannin content of C. ladanifer and C. populifolius leaves aqueous extracts were determined and their antioxidant and antimicrobial activity were fully studied by several in vitro assays. Their major compounds were identified and quantitated by high-performance liquid chromatography with diode array detection coupled to electrospray ion-trap mass spectrometry. Cytotoxicity on a panel of human cancer cells was also determined. C. populifolius extract was stronger antioxidant than C. ladanifer extract in electron transfer reaction based assays but C. ladanifer extract was more effective to inhibit peroxyl radicals. The major compounds in both extracts were ellagitannins, especially punicalagins derivatives, showing C. populifolius a higher content. C. ladanifer showed noteworthy antibacterial activity against Staphylococcus aureus, whereas C. populifolius was effective against Escherichia coli, with MICs values of 154 and 123 microg/mL, respectively. Last, both extracts showed a notorious capacity to inhibit the proliferation of M220 pancreatic cancer cells and MCF7/HER2 and JIMT-1 breast cancer cells. The leaves of these plants suppose a source for water-soluble ellagitannins-enriched polyphenolic extracts with antioxidant and antimicrobial activities. Their cytotoxic activity against several cancer cells may deserve further attention. PMID:20510328

Barrajón-Catalán, Enrique; Fernández-Arroyo, Salvador; Saura, Domingo; Guillén, Emilio; Fernández-Gutiérrez, Alberto; Segura-Carretero, Antonio; Micol, Vicente



Males and females show differential brain activation to taste when hungry and sated in gustatory and reward areas.  


Although males and females differ in eating behavior and prevalence rates for eating disorders and obesity, little is known about gender differences in cortical activation to pleasant and unpleasant pure tastes during the physiological states of hunger and satiety. Twenty-one healthy young adults (12 females and 9 males) underwent two functional magnetic resonance imaging scans. Using four pure tastants of differing qualities (i.e., salty, sour, bitter, sweet), the present study examined gender differences in fMRI activation during two motivational states (hunger and satiety). There was greater change in fMRI activation from hunger to satiety in males than females in response to all tastes within the middle frontal gyrus (BA 10), insula, and cerebellum. Males also had greater change in activation from hunger to satiety, relative to females, in limbic regions including dorsal striatum, amygdala, parahippocampal gyrus, and posterior and anterior cingulate; however, activation was stimulus dependent, despite equivalent ratings in perceived pleasantness and intensity. Interestingly, males and females showed significant change from hunger to satiety in response to citric acid, suggesting that in addition to gender and physiological condition, stimulus quality is an important factor in taste fMRI activation. These gender differences may have implications for the pathophysiology of eating disorders and obesity. PMID:21718731

Haase, Lori; Green, Erin; Murphy, Claire



Dillapiole, Isolated from Peperomia pellucida, Shows Gastroprotector Activity against Ethanol-Induced Gastric Lesions in Wistar Rats.  


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



Syzygium jambos and Solanum guaraniticum show similar antioxidant properties but induce different enzymatic activities in the brain of rats.  


Syzygium jambos and Solanum guaraniticum are both employed in Brazil as medicinal plants, even though their potential toxicity is not well established and they are frequently misused. The aim of this study was investigate the effect of the aqueous leaf extracts of both plants on ?-aminolevulinate dehydratase (?-ALA-D) and acetylcholinesterase (AChE) activities and the antioxidant action against oxidative damage induced by sodium nitroprusside in rats, using in vitro assays. In addition, the presence of gallic, caffeic and chlorogenic acids, as well as rutin, quercetin and kaempferol as bioactive compounds in the extracts was identified by HPLC and their levels quantified. The antioxidant activities of both extracts were assessed by their capabilities to scavenge nitric oxide and to inhibit lipid peroxidation. Only Syzygium jambos presented thiol-peroxidase-like activity. Although neither extract affected the AChE activity, the aqueous extract of Solanum guaraniticum inhibited brain ?-ALA-D activity, suggesting a possible impairment effect on the central nervous system. Our results showed that both extracts exhibited efficient free radical scavenger activity and are an interesting source of bioactive compounds, justifying their use in folk medicine, although Solanum guaraniticum extract could have neurotoxicity properties and we therefore suggest that its use should be restricted to ensure the health of the population. PMID:23912272

Bonfanti, Gabriela; Bitencourt, Paula Rodrigues; Bona, Karine Santos de; Silva, Priscila Sabino da; Jantsch, Letícia B; Pigatto, Aline S; Boligon, Aline; Athayde, Margareth L; Gonçalves, Thissiane L; Moretto, Maria Beatriz



Males and females show differential brain activation to taste when hungry and sated in gustatory and reward areas  

PubMed Central

Although males and females differ in eating behavior and prevalence rates for eating disorders and obesity, little is known about gender differences in cortical activation to pleasant and unpleasant pure tastes during the physiological states of hunger and satiety. Twenty-one healthy young adults (12 females and 9 males) underwent two functional magnetic resonance imaging scans. Using four pure tastants of differing qualities (i.e., salty, sour, bitter, sweet), the present study examined gender differences in fMRI activation during two motivational states (hunger and satiety). There was greater change in fMRI activation from hunger to satiety in males than females in response to all tastes within the middle frontal gyrus (BA 10), insula, and cerebellum. Males also had greater change in activation from hunger to satiety, relative to females, in limbic regions including dorsal striatum, amygdala, parahippocampal gyrus, and posterior and anterior cingulate; however, activation was stimulus dependent, despite equivalent ratings in perceived pleasantness and intensity. Interestingly, males and females showed significant change from hunger to satiety in response to citric acid, suggesting that in addition to gender and physiological condition, stimulus quality is an important factor in taste fMRI activation. These gender differences may have implications for the pathophysiology of eating disorders and obesity.

Haase, Lori; Green, Erin; Murphy, Claire



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


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



Selective imidazoline I2 ligands do not show antidepressant-like activity in the forced swim test in mice.  


Clonidine is an adrenergic agonist with high affinity for alpha2 adrenoceptors that also has affinity for imidazoline receptors. Clonidine has previously been shown to reduce immobility in the forced swim test (FST) in mice. In the present study, this effect was blocked by idazoxan (0.06 mg/kg s.c.) and by yohimbine (1.0 mg/kg s.c.) suggesting that clonidine's effects in this test are mediated via its action at alpha2 sites. Imidazoline I2 site ligands have been shown to inhibit monoamine oxidase and thus may also have antidepressant activity. Three compounds with selective affinity for I2 receptors (BU224, BU239, BDF 8082) were also tested in the FST. These compounds showed no activity either alone or in combination with a subthreshold dose of imipramine in the FST. These results suggest that I2 receptor ligands do not show antidepressant-like activity in the FST in mice. Furthermore the activity of the mixed alpha2/I1 agonist clonidine is most likely to be due to its action at alpha2 sites. PMID:11277603

O'Neill, M F; Osborne, D J; Woodhouse, S M; Conway, M W



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

SciTech Connect

The authors have previously isolated mutants of Escherichia coli which show increased oxidation of heterocyclic furan and thiophene substrates. They have now found that strains carrying the thdA mutation express a novel enzyme activity which oxidizes a variety of substrates containing a sulfone (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.

Juhl, M.J.; Clark, D.P. (Southern Illinois Univ., Carbondale (USA))



Shadow Show  

NSDL National Science Digital Library

This interactive game challenges learners to create shadow shapes utilizing the two-dimensional shapes provided. Players use their knowledge of shapes and spatial sense to determine what individual pieces fit together to match the image. The activity is part of the PBS Kids series "Sid, the Science Kid."



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



Children undergoing cardiac surgery for complex cardiac defects show imbalance between pro- and anti-thrombotic activity  

PubMed Central

Introduction Cardiac surgery with cardiopulmonary bypass (CPB) is associated with the activation of inflammatory mediators that possess prothrombotic activity and could cause postoperative haemostatic disorders. This study was conducted to investigate the effect of cardiac surgery on prothrombotic activity in children undergoing cardiac surgery for complex cardiac defects. Methods Eighteen children (ages 3 to 163 months) undergoing univentricular palliation with total cavopulmonary connection (TCPC) (n = 10) or a biventricular repair (n = 8) for complex cardiac defects were studied. Prothrombotic activity was evaluated by measuring plasma levels of prothrombin fragment 1+2 (F1+2), thromboxane B2 (TxB2), and monocyte chemoattractant protein-1 (MCP-1). Anti-thrombotic activity was evaluated by measuring levels of tissue factor pathway inhibitor (TFPI) before, during, and after cardiac surgery. Results In all patients, cardiac surgery was associated with a significant but transient increase of F1+2, TxB2, TFPI, and MCP-1. Maximal values of F1+2, TxB2, and MCP-1 were found at the end of CPB. In contrast, maximal levels of TFPI were observed at the beginning of CPB. Concentrations of F1+2 at the end of CPB correlated negatively with the minimal oesophageal temperature during CPB. Markers of prothrombotic activity returned to preoperative values from the first postoperative day on. Early postoperative TFPI levels were significantly lower and TxB2 levels significantly higher in patients with TCPC than in those with biventricular repair. Thromboembolic events were not observed. Conclusion Our data suggest that children with complex cardiac defects undergoing cardiac surgery show profound but transient imbalance between pro- and anti-thrombotic activity, which could lead to thromboembolic complications. These alterations are more important after TCPC than after biventricular repair but seem to be determined mainly by low antithrombin III.

Heying, Ruth; van Oeveren, Wim; Wilhelm, Stefanie; Schumacher, Katharina; Grabitz, Ralph G; Messmer, Bruno J; Seghaye, Marie-Christine



Activity map of the tammar X chromosome shows that marsupial X inactivation is incomplete and escape is stochastic  

PubMed Central

Background X chromosome inactivation is a spectacular example of epigenetic silencing. In order to deduce how this complex system evolved, we examined X inactivation in a model marsupial, the tammar wallaby (Macropus eugenii). In marsupials, X inactivation is known to be paternal, incomplete and tissue-specific, and occurs in the absence of an XIST orthologue. Results We examined expression of X-borne genes using quantitative PCR, revealing a range of dosage compensation for different loci. To assess the frequency of 1X- or 2X-active fibroblasts, we investigated expression of 32 X-borne genes at the cellular level using RNA-FISH. In female fibroblasts, two-color RNA-FISH showed that genes were coordinately expressed from the same X (active X) in nuclei in which both loci were inactivated. However, loci on the other X escape inactivation independently, with each locus showing a characteristic frequency of 1X-active and 2X-active nuclei, equivalent to stochastic escape. We constructed an activity map of the tammar wallaby inactive X chromosome, which identified no relationship between gene location and extent of inactivation, nor any correlation with the presence or absence of a Y-borne paralog. Conclusions In the tammar wallaby, one X (presumed to be maternal) is expressed in all cells, but genes on the other (paternal) X escape inactivation independently and at characteristic frequencies. The paternal and incomplete X chromosome inactivation in marsupials, with stochastic escape, appears to be quite distinct from the X chromosome inactivation process in eutherians. We find no evidence for a polar spread of inactivation from an X inactivation center.



Characterization of an antimicrobial peptide produced by Bacillus subtilis subsp. spizezinii showing inhibitory activity towards Haemophilus parasuis.  


Haemophilus parasuis is the pathogen that causes Glässer's disease, a major illness affecting young pigs. The aim of this work was to investigate the antagonistic activity of antimicrobial substances produced by Bacillus species against H. parasuis. Among the tested strains, only Bacillus subtilis ATCC 6633 inhibited H. parasuis growth. The antibacterial substance was purified by ammonium sulfate precipitation, gel filtration chromatography on Sephadex G-50 and ion-exchange chromatography on DEAE-cellulose. The purification was about 100-fold with a yield of 0.33 %. The purified substance was resistant up to 80 °C and pH ranging 3-7, but the substance lost its activity when it was treated with proteases. The peptide had a molecular mass of 1083 Da and its sequence was determined by MS as NRWCFAGDD, which showed no homology with other known antimicrobial peptides. The complete inhibition of H. parasuis growth was observed at 20 µg peptide ml(-1) after 20 min of exposure. The peptide obtained by chemical synthesis also showed antimicrobial activity on H. parasuis. The identification of antimicrobial substances that can be effective against H. parasuis is very relevant to combat this pathogen that causes important losses in swine production. PMID:23519163

Teixeira, Mário Lettieri; Dalla Rosa, Andréia; Brandelli, Adriano



"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



Entorhinal stellate cells show preferred spike phase-locking to theta inputs that is enhanced by correlations in synaptic activity  

PubMed Central

In active networks, excitatory and inhibitory synaptic inputs generate membrane voltage fluctuations that drive spike activity in a probabilistic manner. Despite this, some cells in vivo show a strong propensity to precisely lock to the local field potential and maintain a specific spike-phase relationship relative to other cells. In recordings from rat medial entorhinal cortical stellate cells, we measured spike phase-locking in response to sinusoidal “test” inputs in the presence of different forms of background membrane voltage fluctuations, generated via dynamic clamp. We find that stellate cells show strong and robust spike phase-locking to theta (4–12 Hz) inputs. This response occurs under a wide variety of background membrane voltage fluctuation conditions that include a substantial increase in overall membrane conductance. Furthermore, the IH current present in stellate cells is critical to the enhanced spike phase-locking response at theta. Finally, we show that correlations between inhibitory and excitatory conductance fluctuations, which can arise through feed-back and feed-forward inhibition, can substantially enhance the spike phase-locking response. The enhancement in locking is a result of a selective reduction in the size of low frequency membrane voltage fluctuations due to cancelation of inhibitory and excitatory current fluctuations with correlations. Hence, our results demonstrate that stellate cells have a strong preference for spike phase-locking to theta band inputs and that the absolute magnitude of locking to theta can be modulated by the properties of background membrane voltage fluctuations.

Fernandez, Fernando R.; Malerba, Paola; Bressloff, Paul C.; White, John A.



Synthesis and structure of nanocrystalline TiO2 with lower band gap showing high photocatalytic activity.  


Nanocrystalline TiO2 was synthesized by the solution combustion method using titanyl nitrate and various fuels such as glycine, hexamethylenetetramine, and oxalyldihydrazide. These catalysts are active under visible light, have optical absorption wavelengths below 600 nm, and show superior photocatalytic activity for the degradation of methylene blue and phenol under UV and solar conditions compared to commercial TiO2, Degussa P-25. The higher photocatalytic activity is attributed to the structure of the catalyst. Various studies such as X-ray diffraction, Raman spectroscopy, Brunauer-Emmett-Teller surface area, thermogravimetric-differential thermal analysis, FT-IR spectroscopy, NMR, UV-vis spectroscopy, and surface acidity measurements were conducted. It was concluded that the primary factor for the enhanced activity of combustion-synthesized catalyst is a larger amount of surface hydroxyl groups and a lowered band gap. The lower band gap can be attributed to the carbon inclusion into the TiO2 giving TiO(2-2x)C(x) VO2**. PMID:15835170

Nagaveni, K; Hegde, M S; Ravishankar, N; Subbanna, G N; Madras, Giridhar



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.



Ureido-substituted sulfamates show potent carbonic anhydrase IX inhibitory and antiproliferative activities against breast cancer cell lines.  


A series of 50 sulfamates were obtained by reacting 4-aminophenol with isocyanates followed by sulfamoylation. Most of the new compounds were nanomolar inhibitors of the tumor-associated carbonic anhydrase (CA, EC isoforms IX and XII, whereas they inhibited less cytosolic offtarget isoforms CA I and II. Some of these sulfamates showed significant antiproliferative activity in several breast cancer cell lines, such as SKBR3, MCF10A, ZR75/1, MDA-MB-361 and MCF7, constituting interesting anticancer leads. PMID:22721713

Winum, Jean-Yves; Carta, Fabrizio; Ward, Carol; Mullen, Peter; Harrison, David; Langdon, Simon P; Cecchi, Alessandro; Scozzafava, Andrea; Kunkler, Ian; Supuran, Claudiu T



Origanum vulgare subsp. hirtum Essential Oil Prevented Biofilm Formation and Showed Antibacterial Activity against Planktonic and Sessile Bacterial Cells.  


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



A new chiral 2-(ethylthio)-thiazolone analogue shows strong antitumor activities by inducing cancer cell apoptosis and inhibiting angiogenesis.  


Our initial study showed the potent cytotoxic effects of a series of new chiral 2-(ethylthio)-thiazolone analogues we synthesized. In the present study, we used computer prediction and found that nitro functionality and the modification of substituents R could further improve their activities in the presence of the nitro group. Compound 1s with nitro, naphthyl, ethyl groups, and a chiral center was predicted to be the most effective. We showed that compound 1s could inhibit the growth of five different cancer cell lines in a time-dependent and dose-dependent manner. 1s could induce Hela cell apoptosis by activating the mitochondria apoptotic pathway. In addition, 1s could inhibit the proliferation, migration, tuber formation, and adhesion of human umbilical vein endothelial cells, suggesting its antiangiogenesis effects. Furthermore, we confirmed the in-vivo antitumor effects of 1s on sarcoma S-180-bearing mice. Taken together, chiral 2-(ethylthio)-thiazolone analogue 1s is a promising compound for further anticancer drug development. PMID:22614105

Song, Hongjin; Kai, Ming; Liu, Xiaodong; Yin, Wen; Zeng, Qian; Yao, Xiaojun; Wang, Rui



Substrate uptake tests and quantitative FISH show differences in kinetic growth of bulking and non-bulking activated sludge.  


The competition between filaments and floc formers in activated sludge has been historically described using kinetic selection. However, recent studies have suggested that bacterial storage may also be an important factor in microbial selection, since the dynamic nature of substrate flows into wastewater treatment plants elicit transient responses from microorganisms. Respirometry-based kinetic selection should thus be reevaluated by considering cell storage, and a more reliable method should be developed to include bacterial storage in the analysis of growth of filaments and floc formers in activated sludge. In this study, we applied substrate uptake tests combined with metabolic modeling to determine the growth rates, yields and maintenance coefficients of bulking and non-bulking activated sludge developed in lab scale reactors under feast and famine conditions. The results of quantitative fluorescence in situ hybridization (FISH) showed that the filaments Eikelboom Type 1851, Type 021N, and Thiothrix nivea were dominant in bulking sludge, comprising 42.0 % of mixed liquor volatile suspended solids (MLVSS), with 61.6% of the total filament length extending from flocs into bulk solution. Only low levels of Type 1851 filament length (4.9% of MLVSS) occurred in non-bulking sludge, 83.0% of which grew inside the flocs. The kinetic parameters determined from the substrate uptake tests were consistent with those from respirometry and showed that filamentous bulking sludge had lower growth rates and maintenance coefficients than non-bulking sludge. These results provide support for growth kinetic differences in explaining the competitive strategy of filamentous bacteria. PMID:16155949

Lou, Inchio; de Los Reyes, Francis L



Isolation of 4-coumarate Co-A ligase gene promoter from loblolly pine (Pinus taeda) and characterization of tissue-specific activity in transgenic tobacco.  


We characterized promoter activity of a phenylpropanoid biosynthetic gene encoding 4-coumarate Co-A ligase (4CL), Pta4Clalpha, from Pinus taeda. Histochemical- and quantitative assays of GUS expression in the vascular tissue were performed using transgenic tobacco plants expressing promoter-GUS reporters. Deletion analysis of the Pta4Clalpha promoter showed that the region -524 to -252, which has two AC elements, controls the high expression levels in ray-parenchyma cells of older tobacco stems. High activity level of the promoter domain of Pta4CLalpha was also detected in the xylem cells under bending stress. DNA-protein complexes were detected in the reactions of the Pta4CLalpha promoter fragments with the nuclear proteins of xylem of P. taeda. The AC elements in the Pta4CLalpha promoter appeared to have individual roles during xylem development that are activated in a coordinated manner in response to stress in transgenic tobacco. PMID:19800807

Osakabe, Yuriko; Osakabe, Keishi; Chiang, Vincent L



Use of ELISA and GUS?transformed strains to study competition between pathogenic and non?pathogenic Fusarium oxysporum for root colonization  

Microsoft Academic Search

To characterize the ability of different strains of Fusarium oxysporum to colonize roots, and to analyze competition for root colonization between pathogenic and non?pathogenic strains of F. oxysporum, it was necessary to develop specific labelling techniques for quantification of root colonization. Two methods were selected: the production of polyclonal antibodies, and the use of GUS?transformed strains of F. oxysporum. The

A. Eparvier; C. Alabouvette



New unusual iridoids from the leaves of noni ( Morinda citrifolia L.) show inhibitory effect on ultraviolet B-induced transcriptional activator protein-1 (AP1) activity  

Microsoft Academic Search

A novel iridoid dimer in whose structure the two iridoid units are connected by a rare ether group, together with two new unusual iridoids showing significant inhibition of UVB-induced Activator Protein-1 (AP-1) activity in cell cultures, have been isolated from the leaves of noni (Morinda citrifolia L.). Their structures were determined on the basis of detailed high-field 1D and 2D

Shengmin Sang; Guangming Liu; Kan He; Nanqun Zhu; Zigang Dong; Qunyi Zheng; Robert T Rosen; Chi-Tang Ho



The human interferon-regulated ISG95 protein interacts with RNA polymerase II and shows methyltransferase activity  

SciTech Connect

A major mechanism of cellular resistance to viral invasion involves genes from the interferon signaling pathway, called ISGs (interferon stimulated genes). Global transcriptional profiling studies have linked increased expression of ISG95 (KIAA0082) to response to interferon treatment and viral infection, suggesting that it may be part of the cellular defense against viral replication. In this work, we show that the ISG95 promoter can drive interferon-induced transcription of a reporter gene in Vero cells. Recombinant ISG95 shows RNA- and S-adenosyl-methionine binding and protein methyltransferase activity in vitro. ISG95 interacts with the C-terminal domain of RNA polymerase II, which is consistent with its nuclear localization and with the predicted function of the WW domain found in the C-terminal region of ISG95. The results presented in this work indicate that ISG95 is part of the interferon response pathway and functions in the pre-mRNA processing events mediated by the C-terminal domain of the RNA polymerase II.

Haline-Vaz, Thais; Lima Silva, Tereza Cristina [Center for Structural Molecular Biology, Brazilian Synchrotron Light Laboratory, LNLS P.O. Box 6192, CEP 13080-970, Campinas SP (Brazil); Zanchin, Nilson I.T. [Center for Structural Molecular Biology, Brazilian Synchrotron Light Laboratory, LNLS P.O. Box 6192, CEP 13080-970, Campinas SP (Brazil)], E-mail:



Arabidopsis aux1rcr1 mutation alters AUXIN RESISTANT1 targeting and prevents expression of the auxin reporter DR5:GUS in the root apex  

PubMed Central

Multilevel interactions of the plant hormones ethylene and auxin coordinately and synergistically regulate many aspects of plant growth and development. This study isolated the AUXIN RESISTANT1 (AUX1) allele aux1rcr1 (RCR1 for REVERSING CTR1-10 ROOT1) that suppressed the root growth inhibition conferred by the constitutive ethylene-response constitutive triple response1-10 (ctr1-10) allele. The aux1rcr1 mutation resulted from an L126F substitution at loop 2 of the plasma membrane-associated auxin influx carrier protein AUX1. aux1rcr1 and the T-DNA insertion mutant aux1-T were both defective in auxin transport and many aspects of the auxin response. Unexpectedly, expression of the auxin-response reporter DR5:GUS in the root apex was substantially prevented by the aux1rcr1 but not the aux1-T mutation, even in the presence of the wild-type AUX1 allele. Following treatment with the synthetic auxin 1-naphthaleneacetic acid (NAA), DR5:GUS expression in aux1rcr1 and aux1-T occurred mainly in the root apex and mature zone. NAA-induced DR5:GUS expression in the root apex was markedly prevented by ethylene in genotypes with aux1rcr1 but not in aux1-T genotypes and the wild type. The effect of aux1rcr1 on DR5:GUS expression seemed to be associated with AUX1-expressing domains. Green fluorescence protein-fused aux1rcr1 was localized in the cytoplasm and probably not to the plasma membrane, indicating important roles of the Lys126 residue at loop 2 in AUX1 targeting. The possible effects of aux1rcr1 on DR5:GUS expression are discussed.

Wen, Chi-Kuang



Arabidopsis aux1rcr1 mutation alters AUXIN RESISTANT1 targeting and prevents expression of the auxin reporter DR5:GUS in the root apex.  


Multilevel interactions of the plant hormones ethylene and auxin coordinately and synergistically regulate many aspects of plant growth and development. This study isolated the AUXIN RESISTANT1 (AUX1) allele aux1(rcr1) (RCR1 for REVERSING CTR1-10 ROOT1) that suppressed the root growth inhibition conferred by the constitutive ethylene-response constitutive triple response1-10 (ctr1-10) allele. The aux1(rcr1) mutation resulted from an L126F substitution at loop 2 of the plasma membrane-associated auxin influx carrier protein AUX1. aux1(rcr1) and the T-DNA insertion mutant aux1-T were both defective in auxin transport and many aspects of the auxin response. Unexpectedly, expression of the auxin-response reporter DR5:GUS in the root apex was substantially prevented by the aux1(rcr1) but not the aux1-T mutation, even in the presence of the wild-type AUX1 allele. Following treatment with the synthetic auxin 1-naphthaleneacetic acid (NAA), DR5:GUS expression in aux1(rcr1) and aux1-T occurred mainly in the root apex and mature zone. NAA-induced DR5:GUS expression in the root apex was markedly prevented by ethylene in genotypes with aux1(rcr1) but not in aux1-T genotypes and the wild type. The effect of aux1(rcr1) on DR5:GUS expression seemed to be associated with AUX1-expressing domains. Green fluorescence protein-fused aux1(rcr1) was localized in the cytoplasm and probably not to the plasma membrane, indicating important roles of the Lys(126) residue at loop 2 in AUX1 targeting. The possible effects of aux1(rcr1) on DR5:GUS expression are discussed. PMID:23293348

Yu, Jing; Wen, Chi-Kuang



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


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



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.



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.



Chalcone Synthase Promoters in Petunia Are Active in Pigmented and Unpigmented Cell Types.  

PubMed Central

Chalcone synthase (CHS) catalyzes the first step in the biosynthesis of flavonoids that function in flower pigmentation, protection against stress, and induction of nodulation. The petunia genome contains eight complete chs genes, of which four are differentially expressed in floral tissues and UV-light-induced seedlings. The 5[prime]-flanking regions of these four chs genes were fused to the [beta]-glucuronidase (GUS) reporter gene and introduced into petunia plants by Agrobacterium-mediated transformation. We show that expression of each construct is identical to the expression of the authentic chs gene, implying that the differences in expression pattern between these chs genes are caused at least in part by their promoters. Histochemical analyses of GUS expression show that chs promoters are not only active in pigmented cell types (epidermal cells of the flower corolla and tube and [sub] epidermal cells of the flower stem) but also in a number of unpigmented cell types (mesophylic cells of the corolla, several cell types in the ovary and the seed coat). Comparison of chs-GUS expression and flavonoid accumulation patterns in anthers suggests that intercellular transport of flavonoids and enzymes occurs in this organ. Analysis of the flavonoids accumulated in tissues from mutant lines shows that only a subset of the genes that control flavonoid biosynthesis in the flower operates in the ovary and seed. This implies that (genetic) control of flavonoid biosynthesis is highly tissue specific.

Koes, RE; Van Blokland, R; Quattrocchio, F; Van Tunen, AJ; Mol, J



Meristem, cell division and S phase-dependent activity of wheat histone H4 promoter in transgenic maize plants  

Microsoft Academic Search

We have studied the activity of a wheat histone H4 promoter in transgenic maize plants produced by direct DNA uptake into embryogenic cell suspension protoplasts. Expression patterns revealed by fluorimetric as well as histochemical GUS reporter enzyme assays and Northern hybridisation indicated the cell division-dependent expression of the GUS gene driven by the histone H4 promoter in proliferating cells and

Metin Bilgin; Damla Dedeo?lu; Serik Omirulleh; Adrian Peres; Gilbert Engler; Dirk Inzé; Dénes Dudits; Attila Fehér



Children undergoing cardiac surgery for complex cardiac defects show imbalance between pro- and anti-thrombotic activity  

Microsoft Academic Search

INTRODUCTION: Cardiac surgery with cardiopulmonary bypass (CPB) is associated with the activation of inflammatory mediators that possess prothrombotic activity and could cause postoperative haemostatic disorders. This study was conducted to investigate the effect of cardiac surgery on prothrombotic activity in children undergoing cardiac surgery for complex cardiac defects. METHODS: Eighteen children (ages 3 to 163 months) undergoing univentricular palliation with

Ruth Heying; Wim van Oeveren; Stefanie Wilhelm; Katharina Schumacher; Ralph G Grabitz; Bruno J Messmer; Marie-Christine Seghaye



Active Learning Facilitated by Using a Game-Show Format or Who Doesn't Want to Be a Millionaire?  

ERIC Educational Resources Information Center

|University faculty are increasingly called on to be less of a sage on the stage and more a guide on the side. This discussion introduces the underlying philosophy and assumptions of active learning theory. With this shift in pedagogical philosophy, there has been an increasing call for tools that actively engage students in the learning process.…

Sarason, Yolanda; Banbury, Catherine



Physical Activity in the Life of a Woman with Severe Cerebral Palsy: Showing Competence and Being Socially Connected  

ERIC Educational Resources Information Center

We used a life-history approach to investigate the meanings and experiences of physical activity in the life of a 25-year-old woman with severe cerebral palsy (Amy). Amy and her mother were interviewed about Amy's life and her involvement in physical activity. The conversation was audio-recorded and transcribed verbatim. We interpreted Amy's story…

Gaskin, Cadeyrn J.; Andersen, Mark B.; Morris, Tony



Quantitative assessment of in planta distribution of metabolic activity and gene expression of an endophytic fungus.  


Using perennial ryegrass infected with an Acremonium transformant carrying the Escherichia coli beta-D-glucuronidase gene (gusA) (GUS system) under control of a constitutive promoter, we have developed methods for the quantitative extraction of endophyte-associated GUS activity from plant material. Fluorometric assays of these extracts allow quantitative assessment of the distribution of endophyte-associated GUS activity within single plants (tillers) with high resolution. Fluorescence microscopy with the dye Imagene Green can in addition visualize individual GUS-expressing hyphae. Since the transformant expresses the GUS gene constitutively, GUS activity can be used as an indicator of in planta endophyte metabolic activity. Using this approach we found that (i) the concentration of endophyte metabolic activity in plant tissue decreases with increasing plant size, (ii) approximately 70% of endophyte metabolic activity present in a plant is located in the leaf sheaths, (iii) basal-apical gradients and lateral (younger to older tissue) gradients of endophyte metabolic activity exist and (iv) basal-apical gradients are established early in leaf development. Our data suggest that the concentration of endophyte in each part of the plant is regulated so that a predetermined threshold of total endophyte activity per plant is not exceeded and a consistent distribution pattern is maintained. PMID:10627142

Herd, S; Christensen, M J; Saunders, K; Scott, D B; Schmid, J



Dimeric cyclohexane-1,3-dione oximes inhibit wheat acetyl-CoA carboxylase and show anti-malarial activity.  


A series of dimeric 1,3-cyclohexanedione oxime ethers were synthesized and found to have significant antiplasmodial activity with IC(50)'s in the range 3-12 microM. The most active dimer was tested in the Plasmodium berghei mouse model of malaria and at a dose of 48 mg/kg gave a 45% reduction in parasitaemia. Several commercial herbicides, all known to be inhibitors of maize acetyl-CoA carboxylase, were also tested for antimalarial activity, but were essentially inactive with the exception of butroxydim which gave an IC(50) of 10 microM. PMID:20580556

Louie, Theola; Goodman, C Dean; Holloway, Georgina A; McFadden, Geoffrey I; Mollard, Vanessa; Watson, Keith G



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



In vitro analysis of albendazole sulfoxide enantiomers shows that (+)-(R)-albendazole sulfoxide is the active enantiomer against Taenia solium.  


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. PMID:23229490

Paredes, Adriana; de Campos Lourenço, Tiago; Marzal, Miguel; Rivera, Andrea; Dorny, Pierre; Mahanty, Siddhartha; Guerra-Giraldez, Cristina; García, Hector H; Nash, Theodore E; Cass, Quezia B



Naturally occurring phenolic antibacterial compounds show effectiveness against oral bacteria by a quantitative structure-activity relationship study.  


Natural and synthetic phenolic compounds were evaluated against oral bacteria. A quantitative structure-active relationship approach was applied to the germ-kill activity for a range of phenolic compounds. The lipophilicity and steric effects were found to be two key factors in determining germ-kill activity. The optimum lipophilicity, measured by the logarithm of the octanol/water partition coefficient, or log P, was found to be 5.5 for Fusobacterium nucleatum , a Gram-negative type of oral bacteria that causes bad breath. The optimum log P was found to be 7.9 for Streptococcus mutans , a Gram-positive type of oral bacteria that causes tooth decay. The steric effect of substituents ortho to the phenolic group was found to be critical in reducing antibacterial activity despite having increased lipid solubility approaching the optimum lipophilicity value. The antibacterial activity of phenolic compounds is likely exerted by multiple functions, primarily comes from its ability to act as a nonionic surface-active agent therefore disrupting the lipid-protein interface. PMID:19007234

Greenberg, Michael; Dodds, Michael; Tian, Minmin



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.



Guanine nucleotide binding sites, responsible for adenylate cyclase activation and carbamylcholine binding inhibition, show similar properties in rat heart membranes.  


1/ In rat heart membranes, muscarinic receptors were shown to interact with guanine nucleotide binding sites closely related, if not identical, to those activating adenylate cyclase. The dose-effect curves of GTP, p[NH]ppG, and GTP gamma S for inhibition of carbamylcholine binding (measured by competition with [3H]QNB) and for adenylate cyclase activation (measured in the presence of isoproterenol) were parallel, at both 25 degrees C and 37 degrees C. 2/ Persistent activation of adenylate cyclase was obtained in heart membranes preincubated with p[NH]ppG or GTP gamma S then washed. The affinity for carbamylcholine was reduced after this pretreatment. The SO.5 of p[NH]ppG and GTP gamma S provoking persistent activation of adenylate cyclase and persistent inhibition of carbamylcholine binding were identical. Persistent inhibition of carbamylcholine binding was not additive with the inhibition observed when fresh nucleotide was added after washing. With p[NH]ppG, SO.5 values were unaffected by washing. With GTP gamma S, the SO.5 value for persistent activation of adenylate cyclase (i.e. after washing) and 330 times higher than that implementing activation (i.e. before washing). A similar change was observed when testing the SO.5 of GTP gamma S inhibition of carbamylcholine binding. This might reflect a partial release of GTP gamma S (but not of p[NH]ppG) from "spare" nucleotide binding sites during the washing period. 3/ Adenylate cyclase activity after maximal persistent activation was not increased when 0.1 mM guanine nucleotide, with or without 10 muM isoproterenol, was added to the incubation medium. In contrast, carbamylcholine binding was further decreased when fresh guanine nucleotide was added to the binding assay. This suggests that the proportion of "spare" nucleotide binding sites capable of activating the adenylate cyclase was higher than that capable to inhibit carbamylcholine binding, or that a second class of nucleotide binding sites (binding p[NH]ppG and GTP gamma S reversibly) was also able to inhibit carbamylcholine binding. PMID:7287969

Waelbroeck, M; Robberecht, P; Chatelain, P; Christophe, J



Genetic transformation of peanut ( Arachis hypogaea L.) using cotyledonary node as explant and a promoterless gus::npt II fusion gene based vector  

Microsoft Academic Search

We have generated putative promoter tagged transgenic lines inArachis hypogaea cv JL-24 using cotyledonary node (CN) as an explant and a promoterless gus::nptII bifunctional fusion gene mediated byAgrobacterium transformation. MS medium fortified with 6-benzylaminopurine (BAP) at 4 mg\\/l in combination with 0.1 mg\\/l ?-napthaleneacetic\\u000a acid (NAA) was the most effective out of the various BAP and NAA combinations tested in

T. Swathi Anuradha; S. K. Jami; R. S. Datla; P. B. Kirti



Tissue specificity and expression level of gusA under rolD, mannopine synthase and translation elongation factor 1 subunit a promoters in transgenic Gladiolus plants  

Microsoft Academic Search

Transgenic plants of Gladiolus cv. Jenny Lee were developed that contain the bargusA fusion gene under either the mannopine synthase 2 (mas2), translation elongation factor 1 subunit ? (EF-1?), rolD, or the cauliflower mosaic virus 35S (CaMV 35S) promoters. The relative level of gusA expression in leaves of five to ten independently transformed, in-vitro-grown plants representing each promoter was similar

K. Kamo; A. Blowers



Combined Targeting of Endothelin A Receptor and Epidermal Growth Factor Receptor in Ovarian Cancer Shows Enhanced Antitumor Activity  

Microsoft Academic Search

Ovarian carcinomas overexpress endothelin A receptors (ETAR) and epidermal growth factor (EGF) receptor (EGFR). In these cells, endothelin-1 (ET-1) triggers mitogenic and invasive signaling pathways that are in part mediated by EGFR trans- activation. Combined targeting of ETAR, by the specific ETAR antagonist ZD4054, and of EGFR by the EGFR inhibitor gefitinib (IRESSA), may offer improvements in ovarian carcinoma treatment.

Laura Rosano; Valeriana Di Castro; Francesca Spinella; Giampaolo Tortora; Maria Rita Nicotra; Pier Giorgio Natali; Anna Bagnato



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



Urocortin-1 and -2 double-deficient mice show robust anxiolytic phenotype and modified serotonergic activity in anxiety circuits  

Microsoft Academic Search

The urocortin (Ucn) family of neuropeptides is suggested to be involved in homeostatic coping mechanisms of the central stress response through the activation of corticotropin-releasing factor receptor type 2 (CRFR2). The neuropeptides, Ucn1 and Ucn2, serve as endogenous ligands for the CRFR2, which is highly expressed by the dorsal raphe serotonergic neurons and is suggested to be involved in regulating

A Neufeld-Cohen; A K Evans; D Getselter; A Spyroglou; A Hill; S Gil; M Tsoory; F Beuschlein; C A Lowry; W Vale; A Chen



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


Schizophrenia Subjects Show Intact Success-Related Neural Activation but Impaired Uncertainty Processing during Decision-Making  

Microsoft Academic Search

Decision-making is a complex process that is important for everyday life. This study examined the effect of the degree of success, and outcome uncertainty, on decision-making and associated neural substrate activation in schizophrenia subjects (SZS) and normal comparison subjects (NCS). A total of 15 subjects with a diagnosis of schizophrenia and 15 age- and education-matched NCS participated in this study.

Martin P Paulus; Lawrence Frank; Gregory G Brown; David L Braff



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


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



Purification and characterization of patagonfibrase, a metalloproteinase showing ?-fibrinogenolytic and hemorrhagic activities, from Philodryas patagoniensis snake venom  

Microsoft Academic Search

Venoms of Colubridae snakes are a rich source of novel compounds, which may have applications in medicine and biochemistry. In the present study, we describe the purification and characterization of a metalloproteinase (patagonfibrase), the first protein to be isolated from Philodryas patagoniensis (Colubridae) snake venom. Patagonfibrase is a single-chain protein, showing a molecular mass of 53,224 Da and an acidic

M. E. Peichoto; P. Teibler; S. P. Mackessy; L. Leiva; O. Acosta; L. R. C. Gonçalves; A. M. Tanaka-Azevedo; M. L. Santoro



Crystallographic analyses of an active HIV-1 ribonuclease H domain show structural features that distinguish it from the inactive form.  


. An active recombinant preparation of the carboxy-terminal ribonuclease H (RNase H) domain of HIV-I reverse transcriptase has produced crystals of several different forms, including a trigonal prism form (P3(1); a = b = 52.03, c = 113.9 A with two molecules per asymmetric unit) and a hexagonal tablet form (P6(2)22 or P6(4)22; a = b = 93.5, c = 74.1 A with one molecule per asymmetric unit). The former appears to be isomorphous with crystals of a similar, but inactive, version of the enzyme that was used for a prior crystal structure determination [Davies, Hostomska, Hostomsky, Jordan & Matthews (1991). Science, 252, 88-95]. We have also obtained a structure solution for this crystal form and have refined it with 2.8 A resolution data (R = 0.216). We report here details of our crystallization studies and some initial structural results that verify that the preparation of active HIV-1 RNase H yields a protein that is not just enzymatically, but also structurally, distinguishable from the inactive form. Evidence suggests that region 538-542, which may be involved in the catalytic site and which is disordered in both molecules in the prior structure determination, is ordered in the crystal structure of the active enzyme, although the ordering may include more than one conformation for this loop. It should also be noted that, in the crystal structure of the trigonal form, RNase H monomers associate to form noncrystallographic twofold-symmetric dimers by fusing five-stranded mixed beta sheets into a single ten-stranded dimerwide sheet, an assembly that was not remarked upon by previous investigators. PMID:15299518

Chattopadhyay, D; Finzel, B C; Munson, S H; Evans, D B; Sharma, S K; Strakalaitus, N A; Brunner, D P; Eckenrode, F M; Dauter, Z; Betzel, C; Einspahr, H M



Mono- and dinuclear manganese(III) complexes showing efficient catechol oxidase activity: syntheses, characterization and spectroscopic studies.  


Four side-off compartmental ligands L1-L4 [L1 = N,N'-ethylenebis(3-formyl-5-methyl-salicylaldimine), L2 = N,N'-1-methylethylenebis(3-formyl-5-methylsalicylaldimine), L3 = N,N'-1,1-dimethylethylenebis(3-formyl-5-methylsalicylaldimine) and L4= N,N'-cyclohexenebis(3-formyl-5-methylsalicylaldimine)] having two binding sites, N2O2 and O4, have been chosen to synthesize mononuclear and dinuclear manganese(III) complexes with the aim to study their catecholase activity using 3,5-di-tert-butylcatechol (3,5-DTBC) as substrate in the presence of molecular oxygen. In all cases only mononuclear manganese complexes (1-4) were obtained, with manganese coordination taking place at the N2O2 binding site only, irrespective of the amount of manganese salt used. All these complexes have been characterized by routine physico-chemical techniques. Complex MnL2Cl.4H2O (2) has further been structurally characterized by X-ray single crystal structure analysis. Four dinuclear manganese complexes, 5-8, were obtained after condensing the two pending formyl groups on each ligand (L1-L4) with aniline followed by reaction with MnCl2 to put the second Mn atom onto another N2O2 site. The catalytic activity of all complexes 1-8 has been investigated following the oxidation of 3,5-di-tert-butylcatechol (3,5-DTBC) to 3,5-di-tert-butylbenzoquinone (3,5-DTBQ) with molecular oxygen in two different solvents, methanol and acetonitrile. The study reveals that the catalytic activity is influenced by the solvent and to a significant extent by the backbone of the diamine and the behavior seems to be related mainly to steric rather than electronic factors. Experimental data suggest that a correlation, the lower the E(1/2) value the higher the catalytic activity, can be drawn between E(1/2) and Vmax of the complexes in a particular solvent. The EPR measurements suggest that the catalytic property of the complexes is related to the metal center(s) participation rather than to a radical mechanism. PMID:19809751

Banu, Kazi Sabnam; Chattopadhyay, Tanmay; Banerjee, Arpita; Mukherjee, Madhuparna; Bhattacharya, Santanu; Patra, Goutam Kumar; Zangrando, Ennio; Das, Debasis



Rats with congenital learned helplessness respond less to sucrose but show no deficits in activity or learning  

Microsoft Academic Search

Inbred rat strains for congenital learned helplessness (cLH) and for congenital resistance to learned helplessness (cNLH) were investigated as a model to study genetic predisposition to major depression. Congenitally helpless rats respond less to sucrose under a progressive ratio schedule. This is not confounded by locomotor hypoactivity: in contrast, cLH rats show a slight hyperactivity during the first 5min of

Barbara Vollmayr; Daniel Bachteler; Valentina Vengeliene; Peter Gass; Rainer Spanagel; Fritz Henn



Microglia from Creutzfeldt-Jakob Disease-Infected Brains Are Infectious and Show Specific mRNA Activation Profiles  

Microsoft Academic Search

Neurons are often assumed to be the principal sites for replication of the infectious agents causing Creutzfeldt-Jakob disease (CJD), scrapie, and bovine spongiform encephalopathy because they express high levels of normal and pathological prion protein (PrP). However, isolated brain cell types have not been evaluated for either infection or gene expression. Microglia purified from CJD-infected mice showed infectivity comparable to

Christopher A. Baker; Daniel Martin; Laura Manuelidis



PF2401-SF, standardized fraction of Salvia miltiorrhiza shows anti-inflammatory activity in macrophages and acute arthritis in vivo.  


Standardization of processing methods for herbs is as important as authentication to maintain their quality and ensure their safe use. We had previously prepared a standardized and purified Salvia miltiorrhiza Bunge extract, PF2401-SF, and showed that it protects against liver injury in vivo, at a greater potency than an ethanol extract. PF2401-SF was enriched with tanshinone I (11.5%), tanshinone IIA (41.0%), and cryptotanshinone (19.1%). In this study, we investigated potential anti-inflammatory effects of PF2401-SF in vitro and in vivo. We demonstrated that PF2401-SF shows anti-inflammatory potency on lipopolysaccharide (LPS)-induced nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression in RAW 264.7 cells. A mechanistic study indicated that PF2401-SF induced heme oxygenase (HO)-1 expression through extracellular signal-regulated kinases (ERK1/2) phosphorylation. Moreover, we also evaluated that PF2401-SF significantly reduced inflammation on carrageenan- or dextran-induced acute arthritis in rats. Our results suggest that PF2401-SF may be a potential candidate for the treatment of various inflammatory diseases. PMID:23583334

Jiang, Wen-Yi; Jeon, Byeong-Ho; Kim, Youn-Chul; Lee, Sung Hee; Sohn, Dong Hwan; Seo, Geom Seog



Novel p38? mitogen-activated protein kinase inhibitor shows analgesic efficacy in acute postsurgical dental pain.  


SCIO-469 is a selective p38? mitogen-activated protein kinase (MAPK) inhibitor for preclinical models of acute pain. This prospective, double-blind, randomized clinical study compared efficacy and safety of oral SCIO-469, ibuprofen, and placebo in postsurgical dental pain. Subjects (n = 263) undergoing extraction of 1 or more impacted mandibular third molars received preoperative treatment with SCIO-469 (150, 210, or 300 mg), ibuprofen (400 mg), or placebo; the 210-mg group received 90 mg postoperatively. A 4-point categorical scale and a 100-mm visual analogue scale were used to measure pain intensity. The primary end point was median time from first incision to first rescue medication using the Kaplan-Meier product limit estimator. All SCIO-469 groups had significantly longer times to rescue medication compared with placebo; preoperative and postoperative treatment with 210 + 90 mg SCIO-469 resulted in 8.1 hours versus 4.1 hours to rescue for placebo (P = .003). Ibuprofen also increased time to rescue medication (6.6 hours) versus placebo (P = .04). Dizziness, headache, and nausea were the most frequently reported adverse events. This is the first clinical demonstration of antinociceptive effects in acute pain with preoperative administration of a p38? MAPK inhibitor. PMID:21659629

Tong, Sandra E; Daniels, Stephen E; Black, Peter; Chang, Stephen; Protter, Andrew; Desjardins, Paul J



Agaricus bisporus and Agaricus brasiliensis (1?6)-?-D-glucans show immunostimulatory activity on human THP-1 derived macrophages.  


The (1?6)-?-D-glucans from Agaricus bisporus and Agaricus brasiliensis were purified to evaluate their effects on the innate immune system. THP-1 macrophages were used to investigate the induction of the expression of TNF-?, IL1?, and COX-2 by RT-PCR. The purification of the polysaccharides gave rise to fractions containing 96-98% of glucose. The samples were analyzed by GC-MS, HPSEC and (13)C NMR, which confirmed the presence of homogeneous (1?6)-?-D-glucans. The ?-glucans were incubated with THP-1 derived macrophages, for 3 h and 6 h to evaluate their effects on the expression of pro-inflammatory genes. Both ?-glucans stimulated the expression of such genes as much as the pro-inflammatory control (LPS). When the cells were incubated with LPS+?-glucan, a significant inhibition of the expression of IL-1? and COX-2 was observed for both treatments after 3 h of incubation. By the results, we conclude that the (1?6)-?-D-glucans present an immunostimulatory activity when administered to THP-1 derived macrophages. PMID:23544515

Smiderle, Fhernanda R; Alquini, Giovana; Tadra-Sfeir, Michelle Z; Iacomini, Marcello; Wichers, Harry J; Van Griensven, Leo J L D



Plasminogen Activator Inhibitor-Type I Gene Deficient Mice Show Reduced Influx of Neutrophils in Ventilator-Induced Lung Injury  

PubMed Central

Ventilator-induced lung injury (VILI) is associated with inhibition of the fibrinolytic system secondary to increased production of plasminogen activator inhibitor- (PAI-)1. To determine the role of PAI-1 on pulmonary coagulopathy and inflammation during mechanical ventilation, PAI-1 gene-deficient mice and their wild-type littermates were anesthetized (control), or anesthetized, tracheotomized and subsequently ventilated for 5 hours with either low tidal volumes (LVT) or high tidal volumes (HVT). VILI was assessed by pulmonary coagulopathy, lung wet-to-dry ratios, total protein level in bronchoalveolar lavage fluid, neutrophil influx, histopathology, and pulmonary and plasma cytokine levels. Ventilation resulted in pulmonary coagulopathy and inflammation, with more injury following ventilation with HVT as compared to LVT. In PAI-1 gene-deficient mice, the influx of neutrophils in the pulmonary compartment was attenuated, while increased levels of pulmonary cytokines were found. Other endpoints of VILI were not different between PAI-1 gene-deficient and wild-type mice. These data indicate that a defect fibrinolytic response attenuates recruitment of neutrophils in VILI.

Wolthuis, Esther K.; Vlaar, Alexander P. J.; Hofstra, Jorrit-Jan H.; Roelofs, Joris J. T. H.; de Waard, Vivian; Juffermans, Nicole P.; Schultz, Marcus J.



Crack users show high rates of antisocial personality disorder, engagement in illegal activities and other psychosocial problems.  


The aim of this study was to compare three groups of Brazilian psychoactive substance (PAS) abuse patients (crack cocaine users, cocaine snorters, and non-cocaine PAS users) in terms of psychiatric comorbidities and severity of psychosocial problems. A cross-sectional, multi-center study was conducted at five Brazilian research centers. A total of 738 current PAS abusers seeking specialized treatment (outpatient and inpatient clinics) were assessed using the sixth version of the Addiction Severity Index (ASI-6): 293 patients using crack cocaine were compared with 126 using powder cocaine and 319 using non-cocaine PAS (mostly alcohol and marijuana). Psychiatric comorbidities were assessed in a smaller sample (290 cases), originating from three of the centers, using the Mini International Neuropsychiatric Interview Plus (MINI-Plus). Crack and powder cocaine users were significantly younger than non-cocaine PAS users (31.1 ± 8.1 and 32.9 ± 8.8 vs. 42.4 ± 12, respectively; p < .001). Crack users presented a higher rate of antisocial personality disorder (25%) than powder cocaine (9%) and non-cocaine PAS users (9%), even when adjusted for confounding factors (Pr = 2.6; 95% CI 1.10-6.40). According to ASI-6 summary scores, crack users presented a significantly higher rate of occupational, family, and legal problems and reported more illegal and violent activities such as burglary and theft (23%) and threatening or assaulting (32%) than non-cocaine PAS users. Our findings, combined with the recent increase observed in the prevalence of crack use in Brazil, highlight the severity of psychiatric symptoms and psychosocial problems related to this powerful drug and corroborate the already suggested association between crack/cocaine, violence, and legal problems. Treatment programs for crack users should routinely consider the possibility of associated psychiatric comorbidities, such as antisocial personality disorder, which may affect treatment outcomes. PMID:22691017

Paim Kessler, Felix Henrique; Barbosa Terra, Mauro; Faller, Sibele; Ravy Stolf, Anderson; Carolina Peuker, Ana; Benzano, Daniela; Pechansky, Flavio



The selective VEGFR1-3 inhibitor axitinib (AG-013736) shows antitumor activity in human neuroblastoma xenografts.  


Tumor angiogenesis in childhood neuroblastoma is an important prognostic factor suggesting a potential role for antiangiogenic agents in the treatment of high-risk disease. Within the KidsCancerKinome project, we evaluated the new oral selective pan-VEGFR tyrosine kinase inhibitor axitinib (AG-013736) against neuroblastoma cell lines and the subcutaneous and orthotopic xenograft model IGR-N91 derived from a primary bone marrow metastasis. Axitinib reduced cell proliferation in a dose-dependent manner with IC(50) doses between 274 and >10,000 nmol/l. Oral treatment with 30 mg/kg BID for 2 weeks in advanced tumors yielded significant tumor growth delay, with a median time to reach five times initial tumor volume of 11.4 days compared to controls (p = 0.0006) and resulted in significant reduction in bioluminescence. Simultaneous inhibition of VEGFR downstream effector mTOR using rapamycin 20 mg/kg q2d×5 did not statistically enhance tumor growth delay compared to single agent activities. Axitinib downregulated VEGFR-2 phosphorylation resulting in significantly decreased microvessel density (MVD) and overall surface fraction of tumor vessels (OSFV) in all xenografts as measured by CD34 immunohistochemical staining (mean MVD ± SD and OSFV at 14 days 21.27 ± 10.03 in treated tumors vs. 48.79 ± 17.27 in controls and 0.56% vs. 1.29%; p = 0.0006, respectively). We further explored the effects of axitinib on circulating mature endothelial cells (CECs) and endothelial progenitor cells (CEPs) measured by flow cytometry. While only transient modification was observed for CECs, CEP counts were significantly reduced during and up to 14 days after end of treatment. Axitinib has potent antiangiogenic properties that may warrant further evaluation in neuroblastoma. PMID:20715103

Rössler, Jochen; Monnet, Yann; Farace, Francoise; Opolon, Paule; Daudigeos-Dubus, Estelle; Bourredjem, Abderrahmane; Vassal, Gilles; Geoerger, Birgit



Rats with congenital learned helplessness respond less to sucrose but show no deficits in activity or learning.  


Inbred rat strains for congenital learned helplessness (cLH) and for congenital resistance to learned helplessness (cNLH) were investigated as a model to study genetic predisposition to major depression. Congenitally helpless rats respond less to sucrose under a progressive ratio schedule. This is not confounded by locomotor hypoactivity: in contrast, cLH rats show a slight hyperactivity during the first 5 min of an open field test. cLH rats acquire operant responding to sucrose as readily as cNLH rats and exhibit normal memory acquisition and retrieval in the Morris water maze, thus ruling out general learning deficits as the cause of the decreased response to sucrose. Reduced total responses and reduced breaking points for sucrose in the cLH strain argue for anhedonia, which is an analogue to loss of pleasure essential for the diagnosis of major depressive episodes, and thus confirm the validity of congenitally learned helpless rats as a model of major depression. PMID:15033295

Vollmayr, Barbara; Bachteler, Daniel; Vengeliene, Valentina; Gass, Peter; Spanagel, Rainer; Henn, Fritz



TRAIL shows potential cardioprotective activity  

Microsoft Academic Search

Summary  Recent clinical trials carried out in patients with advanced cancer have shown that recombinant TRAIL administration is usually\\u000a safe and well tolerated when used either alone or in association with chemotherapeutic drugs. Notably, anticancer chemotherapy\\u000a can be associated to cardiomiopathy. We have here demonstrated that TRAIL (administrated as either recombinant soluble TRAIL\\u000a or as AAV-TRAIL expression viral vector) reduced the

Barbara Toffoli; Stella Bernardi; Riccardo Candido; Serena Zacchigna; Bruno Fabris; Paola Secchiero


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



Analysis of Protease Activity in Live Antigen-presenting Cells Shows Regulation of the Phagosomal Proteolytic Contents During Dendritic Cell Activation  

PubMed Central

Here, we describe a new approach designed to monitor the proteolytic activity of maturing phagosomes in live antigen-presenting cells. We find that an ingested particle sequentially encounters distinct protease activities during phagosomal maturation. Incorporation of active proteases into the phagosome of the macrophage cell line J774 indicates that phagosome maturation involves progressive fusion with early and late endocytic compartments. In contrast, phagosome biogenesis in bone marrow–derived dendritic cells (DCs) and macrophages preferentially involves endocytic compartments enriched in cathepsin S. Kinetics of phagosomal maturation is faster in macrophages than in DCs. Furthermore, the delivery of active proteases to the phagosome is significantly reduced after the activation of DCs with lipopolysaccharide. This observation is in agreement with the notion that DCs prevent the premature destruction of antigenic determinants to optimize T cell activation. Phagosomal maturation is therefore a tightly regulated process that varies according to the type and differentiation stage of the phagocyte.

Lennon-Dumenil, Ana-Maria; Bakker, Arnold H.; Maehr, Rene; Fiebiger, Edda; Overkleeft, Herman S.; Rosemblatt, Mario; Ploegh, Hidde L.; Lagaudriere-Gesbert, Cecile



Inactivated Orf Virus Shows Antifibrotic Activity and Inhibits Human Hepatitis B Virus (HBV) and Hepatitis C Virus (HCV) Replication in Preclinical Models  

PubMed Central

Inactivated orf virus (iORFV), strain D1701, is a potent immune modulator in various animal species. We recently demonstrated that iORFV induces strong antiviral activity in animal models of acute and chronic viral infections. In addition, we found D1701-mediated antifibrotic effects in different rat models of liver fibrosis. In the present study, we compare iORFV derived from two different strains of ORFV, D1701 and NZ2, respectively, with respect to their antifibrotic potential as well as their potential to induce an antiviral response controlling infections with the hepatotropic pathogens hepatitis C virus (HCV) and hepatitis B virus (HBV). Both strains of ORFV showed anti-viral activity against HCV in vitro and against HBV in a transgenic mouse model without signs of necro-inflammation in vivo. Our experiments suggest that the absence of liver damage is potentially mediated by iORFV-induced downregulation of antigen cross-presentation in liver sinus endothelial cells. Furthermore, both strains showed significant anti-fibrotic activity in rat models of liver fibrosis. iORFV strain NZ2 appeared more potent compared to strain D1701 with respect to both its antiviral and antifibrotic activity on the basis of dosages estimated by titration of active virus. These results show a potential therapeutic approach against two important human liver pathogens HBV and HCV that independently addresses concomitant liver fibrosis. Further studies are required to characterize the details of the mechanisms involved in this novel therapeutic principle.

Paulsen, Daniela; Urban, Andreas; Knorr, Andreas; Hirth-Dietrich, Claudia; Siegling, Angela; Volk, Hans-Dieter; Mercer, Andrew A.; Limmer, Andreas; Schumak, Beatrix; Knolle, Percy; Ruebsamen-Schaeff, Helga; Weber, Olaf



Plasma membrane of Beta vulgaris storage root shows high water channel activity regulated by cytoplasmic pH and a dual range of calcium concentrations  

Microsoft Academic Search

Plasma membrane vesicles isolated by two-phase par- titioning from the storage root of Beta vulgaris show atypically high water permeability that is equivalent only to those reported for active aquaporins in tono- plast or animal red cells (Pf5542 l ms 21 ). The values were determined from the shrinking kinetics measured by stopped-flow light scattering. This high Pf was only

Karina Alleva; Christa M. Niemietz; Moira Sutka; Christophe Maurel; Mario Parisi; Stephen D. Tyerman; Gabriela Amodeo



What Do Maps Show?  

ERIC Educational Resources Information Center

|This curriculum packet, appropriate for grades 4-8, features a teaching poster which shows different types of maps (different views of Salt Lake City, Utah), as well as three reproducible maps and reproducible activity sheets which complement the maps. The poster provides teacher background, including step-by-step lesson plans for four geography…

Geological Survey (Dept. of Interior), Reston, VA.


ISU Demonstration Road Show  

NSDL National Science Digital Library

The Idaho State University Department of Physics conducts science demonstration shows at SE Idaho schools. Four different presentations are currently available; "Forces and Motion", "States of Matter", "Electricity and Magnetism", and "Sound and Waves". Student activities and descriptions of the demonstrated material are also provided.

Shropshire, Steven



The 5'-flanking region of the RP58 coding sequence shows prominent promoter activity in multipolar cells in the subventricular zone during corticogenesis.  


Pyramidal neurons of the neocortex are produced from progenitor cells located in the neocortical ventricular zone (VZ) and subventricular zone (SVZ) during embryogenesis. RP58 is a transcriptional repressor that is strongly expressed in the developing brain and plays an essential role in corticogenesis. The expression of RP58 is strictly regulated in a time-dependent and spatially restricted manner. It is maximally expressed in E15-16 embryonic cerebral cortex, localized specifically to the cortical plate and SVZ of the neocortex, hippocampus, and parts of amygdala during brain development, and found in glutamatergic but not GABAergic neurons. Identification of the promoter activity underlying specific expression patterns provides important clues to their mechanisms of action. Here, we show that the RP58 gene promoter is activated prominently in multipolar migrating cells, the first in vivo analysis of RP58 promoter activity in the brain. The 5.3 kb 5'-flanking genomic DNA of the RP58 coding region demonstrates promoter activity in neurons both in vitro and in vivo. This promoter is highly responsive to the transcription factor neurogenin2 (Ngn2), which is a direct upstream activator of RP58 expression. Using in utero electroporation, we demonstrate that RP58 gene promoter activity is first detected in a subpopulation of pin-like VZ cells, then prominently activated in migrating multipolar cells in the multipolar cell accumulation zone (MAZ) located just above the VZ. In dissociated primary cultured cortical neurons, RP58 promoter activity mimics in vivo expression patterns from a molecular standpoint that RP58 is expressed in a fraction of Sox2-positive progenitor cells, Ngn2-positive neuronal committed cells, and Tuj1-positive young neurons, but not in Dlx2-positive GABAergic neurons. Finally, we show that Cre recombinase expression under the control of the RP58 gene promoter is a feasible tool for conditional gene switching in post-mitotic multipolar migrating young neurons in the developing cerebral cortex. PMID:22119643

Ohtaka-Maruyama, C; Hirai, S; Miwa, A; Takahashi, A; Okado, H



An alpha2,6-sialyltransferase cloned from Photobacterium leiognathi strain JT-SHIZ-119 shows both sialyltransferase and neuraminidase activity.  


We cloned, expressed, and characterized a novel beta-galactoside alpha2,6-sialyltransferase from Photobacterium leiognathi strain JT-SHIZ-119. The protein showed 56-96% identity to the marine bacterial alpha2,6-sialyltransferases classified into glycosyltransferase family 80. The sialyltransferase activity of the N-terminal truncated form of the recombinant enzyme was 1477 U/L of Escherichia coli culture. The truncated recombinant enzyme was purified as a single band by sodium dodecyl sulfate polyacrylamide gel electrophoresis through 3 column chromatography steps. The enzyme had distinct activity compared with known marine bacterial alpha2,6-sialyltransferases. Although alpha2,6-sialyltransferases cloned from marine bacteria, such as Photobacterium damselae strain JT0160, P. leiognathi strain JT-SHIZ-145, and Photobacterium sp. strain JT-ISH-224, show only alpha2,6-sialyltransferase activity, the recombinant enzyme cloned from P. leiognathi strain JT-SHIZ-119 showed both alpha2,6-sialyltransferase and alpha2,6-linkage-specific neuraminidase activity. Our results provide important information toward a comprehensive understanding of the bacterial sialyltransferases belonging to the group 80 glycosyltransferase family in the CAZy database. PMID:19797322

Mine, Toshiki; Katayama, Sakurako; Kajiwara, Hitomi; Tsunashima, Masako; Tsukamoto, Hiroshi; Takakura, Yoshimitsu; Yamamoto, Takeshi



Nobiletin, a polymethoxylated flavonoid from citrus, shows anti-angiogenic activity in a zebrafish in vivo model and HUVEC in vitro model.  


Traditional Chinese medicinal herbs are a rich source of compounds with reported anti-inflammatory and anti-carcinogenic effects. Growing evidence shows the codependence of chronic inflammation and angiogenesis, and the potential benefits of targeting angiogenesis in the treatment of chronic inflammation and targeting inflammation in the treatment of diseases with impaired angiogenesis. We hypothesized that the anti-inflammatory activity of the natural compounds may owe at least some of its efficacy to their anti-angiogenic activity and hence we investigated the anti-angiogenic activity of these compounds in vivo in zebrafish embryos and in vitro in human umbilical vein endothelial cells (HUVECs). Nobiletin, a polymethoxylated flavonoid from citrus fruits, showed anti-angiogenic activity in both assays. Nobiletin inhibited the formation of intersegmental vessels (ISVs) in live transgenic zebrafish embryos expressing green fluorescent protein (GFP) in the vasculature. Cell cycle analysis of dissociated zebrafish embryo cells showed that nobiletin induced G0/G1 phase accumulation in a dose-dependent manner in GFP-positive endothelial cells. Nobiletin also dose-dependently induced VEGF-A mRNA expression. In HUVECs, nobiletin inhibited endothelial cell proliferation and, to a greater extent, tube formation in a dose-dependent manner. As in the in vivo study, nobiletin induced G0/G1 cell cycle arrest in HUVECs. However, this arrest was not accompanied by an increase in apoptosis, indicating a cytostatic effect of nobiletin. This study, for the first time, identifies nobiletin as having potent anti-angiogenic activity and suggests that nobiletin has a great potential for future research and development as a cytostatic anti-proliferative agent. PMID:21748787

Lam, Kai Heng; Alex, Deepa; Lam, In Kei; Tsui, Stephen Kwok Wing; Yang, Zi Feng; Lee, Simon Ming Yuen



Mx1, Mx2 and Mx3 proteins from the gilthead seabream (Sparus aurata) show in vitro antiviral activity against RNA and DNA viruses.  


Mx proteins are important components of the antiviral innate immune response mediated by type I interferon. Classically, these proteins have been considered to be triggered by viral RNA, thus showing activity against RNA viruses. Actually, three Mx proteins (SauMx1, SauMx2 and SauMx3) from gilthead seabream (Sparus aurata) have previously shown antiviral activity against a dsRNA virus: the infectious pancreatic necrosis virus (IPNV) in vitro. For further characterizing their antiviral spectrum, the activity of SauMx proteins were tested against three different viral pathogens of fish: the lymphocystis disease virus (LCDV, a dsDNA virus), a pathogen of gilthead seabream; the viral haemorrhagic septicaemia virus (VHSV, a ssRNA virus), to which gilthead seabream is considered a reservoir species; and the European sheatfish virus (ESV, a dsDNA virus), that has not been detected in gilthead seabream to date. Three clonal populations of CHSE-214 cells developed in a previous study, stably expressing SauMx1, SauMx2 and SauMx3, respectively, were challenged with the three viruses. Results combining cytopathic effects and virus yield reduction assays showed that SauMx1 protected the cells against VHSV and LCDV, SauMx2 protected against ESV and LCDV, and SauMx3 showed activity only against VHSV. This study, besides confirming the antiviral activity of the three gilthead seabream Mx proteins, is the first report of the protective effect of a fish Mx against DNA viruses. Additionally, it discloses a clear specificity between Mx proteins and virus targets, supporting the idea that the relationship between virus and Mx proteins is finely tuned. PMID:23911421

Fernández-Trujillo, M A; García-Rosado, E; Alonso, M C; Castro, D; Alvarez, M C; Béjar, J



A Utility Accrual Scheduling Algorithm for Real-Time Activities with Mutual Exclusion Resource Constraints  

Microsoft Academic Search

This paper presents a uni-processor real-time scheduling algorithm called the Generic Utility Scheduling algorithm (which we will refer to simply as GUS). GUS solves a previously open real-time scheduling problem|scheduling application activities that have time con- straints specifled using arbitrarily shaped time\\/utility functions, and have mutual exclusion resource constraints. A time\\/utility function is a time constraint speciflcation that describes an

Peng Li; Haisang Wu; Binoy Ravindran; E. Douglas Jensen



Viewing television talk shows  

Microsoft Academic Search

We examined how motivation, audience activity, and attitudes influenced the likelihood of watching societal?issue and relational topics on television talk programs. Path analysis supported differences in ritualized and instrumental motives for watching talk shows. Information and exciting?entertainment motivation predicted greater’ realism of, affinity with, involvement with, and intent to watch talk television. Pass?time motivation predicted reduced affinity with and intent

Alan M. Rubin; Mary M. Step



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



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



Rifamycin SV administered by intra-articular infiltrations shows disease modifying activity in patients with pauci- or polyarticular juvenile rheumatoid arthritis.  


The therapeutic activity of rifamycin SV administered by the intra-articular route was evaluated in 52 children with juvenile rheumatoid arthritis (oligopolyarthritis). Each active joint was injected once a week for 10 weeks; thereafter patients were followed for 3-48 months. The number of active joints and joints with limitation of motion, the erythrocyte sedimentation rate (ESR) and C-reactive protein improved significantly at the end of the treatment cycle, with progressive improvement during the subsequent period of observation. At 48-month of follow-up, 78% of joints did not present signs of inflammation; and 66% of joints showed no functional limitations. Joints without radiological lesions at baseline and large joints responded best to the rifamycin treatment. Persistent knee effusions were reabsorbed completely in most cases during the treatment and within the first 6 months of follow-up. Recurrences of synovitis were observed in 7% of joints. De novo radiological lesions in initially undamaged joints occurred during the second year of follow-up in only 10% of patients. At 24 months, 62% of patients with oligoarthritis and 24% with polyarthritis showed complete remission in all affected joints and recovered movement in all those joints which had shown limitations at baseline. There was also a normalization of inflammatory indexes (ESR, C-reactive protein) and regression of general features of disease. Further long term studies are now required to confirm these promising preliminary results. PMID:8112482

Caruso, I; Principi, N; D'Urbino, G; Santandrea, S; Boccassini, L; Montrone, F; Sarzi Puttini, P C; Bombaci, A; Bozzato, A; Azzolini, V


A dual mTORC1 and mTORC2 inhibitor shows antitumor activity in esophageal squamous cell carcinoma cells and sensitizes them to cisplatin.  


The mammalian target of rapamycin (mTOR) signaling pathway is critical for the growth and proliferation of various malignant tumors, including esophageal squamous cell carcinoma (ESCC). Therefore, targeting of mTOR protein is a promising strategy for therapy in this disease. In the present study, we examined the antitumor effects of a specific mTOR kinase inhibitor, PP242, which blocks both mTOR complex 1 (mTORC1) and mTOR complex 2 (mTORC2) pathways, in two ESCC cell lines: Eca-109 and TE-1. We showed that PP242, but not rapamycin, attenuated the activities of both mTORC1 and mTORC2 signaling in ESCC. PP242 inhibited 4E-binding protein-1 phosphorylation and abrogated mTORC1-dependent PI3K/Akt feedback activation. Significantly, PP242 effectively suppressed ESCC cell proliferation, induced apoptosis, and arrested the cell cycle. Furthermore, PP242 promoted cisplatin-induced apoptosis and enhanced the antitumor efficacy of cisplatin in ESCC cells, which was likely to be associated with inhibition of Akt activity. Our results show that simultaneous targeting of both mTORC1 and mTORC2 pathways leads to effective antitumor actions in ESCC, and strongly suggest that dual mTORC1/2 inhibitors should be developed as potential agents for the treatment of ESCC. PMID:23838676

Huang, Yu; Xi, Qingsong; Chen, Yu; Wang, Jing; Peng, Ping; Xia, Shu; Yu, Shiying



A ruthenium(II) arene complex showing emission enhancement and photocleavage activity towards DNA from singlet and triplet excited states respectively.  


The emission enhancement behavior and photocleavage activity of a ruthenium(II) arene complex, [(?(6)-p-cymene)Ru(dppn)(py)](2+) (1) (dppn = 4,5,9,16-tetraaza-dibenzo[a,c]naphthacene, py = pyridine), towards DNA were compared with [(?(6)-p-cymene)Ru(bpy)(py)](2+) (2), [Ru(bpy)2(dppz)](2+) (3) and [Ru(bpy)2(dppn)](2+) (4) (bpy = 2,2'-bipyridine, dppz = dipyrido-[3,2-a:2',3'-c]phenazine). It was found that 1 emits fluorescence from the dppn-based ligand-centered (LC) singlet excited state and generates singlet oxygen ((1)O2) from the dppn-based LC triplet excited state. As a result, 1 displays emission enhancement behavior and photocleavage activity towards DNA simultaneously. In contrast, 3 is the most classical DNA light switch but shows poor DNA photocleavage activity, while 4 is an efficient DNA photocleaver but cannot report DNA binding by luminescence enhancement. An increased cytotoxicity against human lung carcinoma cells A549 by about 10-fold was also observed for 1 upon visible light activation. These intriguing properties result from the unique combination of the Ru(II) arene and dppn subunits. PMID:23459918

Chen, Yongjie; Lei, Wanhua; Jiang, Guoyu; Zhou, Qianxiong; Hou, Yuanjun; Li, Chao; Zhang, Baowen; Wang, Xuesong



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



Camptothecin analog (CPT-11)-sensitive human pancreatic tumor cell line QGP-1N shows resistance to SN-38, an active metabolite of CPT-11.  


In the course of our study to determine the cross-sensitivity between CPT-11 and its active metabolite, SN-38, we found a SN-38-resistant human pancreatic tumor cell line, QGP-1N, which shows sensitivity to CPT-11. The IC50 of SN-38 was 152 times greater for QGP-1N than for SUIT-2, also a human pancreatic tumor cell line, whose IC50 of CPT-11 was similar to that for QGP-1N. The uptakes of CPT-11 and SN-38 and the intracellular conversion of CPT-11 to SN-38 could not explain the difference in sensitivity. DNA synthesis of QGP-1N cells was inhibited by CPT-11 which did not affect that of SUIT-2, while SN-38 inhibited the DNA synthesis of SUIT-2 at lower concentrations than that of QGP-1N. The inhibition test of topoisomerase I catalytic activity by CPT-11 or SN-38 revealed no difference in the biochemical properties of the topoisomerase I enzymes to the compounds between these two cell lines. These results indicate that CPT-11 should have its own inhibitory effect on DNA synthesis through a yet unknown mechanism in QGP-1N cells, although SN-38 plays an essential role in the antitumor activity of CPT-11 in SUIT-2 cells. In some cases, the antitumor effect of CPT-11 might be consequent not only on SN-38 but also on CPT-11 itself. PMID:1329748

Takeda, S; Shimazoe, T; Kuga, H; Sato, K; Kono, A



Recombinant enzymes overexpressed in bacteria show broad catalytic specificity of human cytochrome P450 2W1 and limited activity of human cytochrome P450 2S1.  


Human cytochromes P450 2S1 and 2W1 have received only limited attention with regard to characterization of function. Both cytochromes P450 have been reported to be overexpressed in human tumors, and cytochrome P450 2S1 is induced by carcinogenic polycyclic hydrocarbons. We report methods for high-level expression and purification of both cytochromes P450 from Escherichia coli, with the goal of establishing function. The level of expression of human cytochrome P450 2W1 achieved using codon optimization for E. coli was 1800 nmol of cytochrome P450 per liter of culture, the highest level achieved in this laboratory to date. Assays with a number of the typical cytochrome P450 substrates showed no detectable activity, including some for which qualitative reports have appeared in the literature. Cytochrome P450 2W1 catalyzed benzphetamine N-demethylation (k(cat), 3.8/min) and arachidonic acid oxidation, albeit at a very low rate (approximately 0.05/min). In a umu genotoxicity screen, cytochrome P450 2W1 catalyzed the activation of several procarcinogens, particularly polycyclic hydrocarbon diols, but cytochrome P450 2S1 did not. The bioactivation of procarcinogens by cytochrome P450 2W1 may be of significance in the context of reports of preferential expression of the enzyme in tumors, in that activation of procarcinogens could lead to the accumulation of mutations and enhance the carcinogenic process. PMID:16551781

Wu, Zhong-Liu; Sohl, Christal D; Shimada, Tsutomu; Guengerich, F Peter



Polysaccharides from the Medicinal Mushroom Cordyceps taii Show Antioxidant and Immunoenhancing Activities in a D-Galactose-Induced Aging Mouse Model  

PubMed Central

Cordyceps taii, an edible medicinal mushroom native to south China, is recognized as an unparalleled resource of healthy foods and drug discovery. In the present study, the antioxidant pharmacological properties of C. taii were systematically investigated. In vitro assays revealed the scavenging activities of the aqueous extract and polysaccharides of C. taii against various free radicals, that is, 1,1-diphenyl-2-picrylhydrazyl radical, hydroxyl radical, and superoxide anion radical. The EC50 values for superoxide anion-free radical ranged from 2.04?mg/mL to 2.49?mg/mL, which was at least 2.6-fold stronger than that of antioxidant thiourea. The polysaccharides also significantly enhanced the antioxidant enzyme activities (superoxide dismutase, catalase, and glutathione peroxidase) and markedly decreased the malondialdehyde production of lipid peroxidation in a D-galactose-induced aging mouse model. Interestingly, the immune function of the administration group was significantly boosted compared with the D-galactose-induced aging model group. Therefore, the C. taii polysaccharides possessed potent antioxidant activity closely associated with immune function enhancement and free radical scavenging. These findings suggest that the polysaccharides are a promising source of natural antioxidants and antiaging drugs. Consequently, a preliminary chemical investigation was performed using gas chromatography-mass spectroscopy and revealed that the polysaccharides studied were mainly composed of glucose, mannose, and galactose. Fourier-transform infrared spectra also showed characteristic polysaccharide absorption bands.

Xiao, Jian-Hui; Xiao, Dai-Min; Chen, Dai-Xiong; Xiao, Yu; Liang, Zong-Qi; Zhong, Jian-Jiang



Activity of the Novel Peptide Arminin against Multiresistant Human Pathogens Shows the Considerable Potential of Phylogenetically Ancient Organisms as Drug Sources?  

PubMed Central

The emergence of multidrug-resistant bacteria highlights the need for new antibacterial agents. Arminin 1a is a novel antimicrobial peptide discovered during investigations of the epithelial defense of the ancient metazoan Hydra. Following proteolytic processing, the 31-amino-acid-long positively charged C-terminal part of arminin 1a exhibits potent and broad-spectrum activity against bacteria, including multiresistant human pathogenic strains, such as methicillin-resistant Staphylococcus aureus (MRSA) strains (minimal bactericidal concentration, 0.4 ?M to 0.8 ?M). Ultrastructural observations indicate that bacteria are killed by disruption of the bacterial cell wall. Remarkably, the antibacterial activity of arminin 1a is not affected under the physiological salt conditions of human blood. In addition, arminin 1a is a selective antibacterial agent that does not affect human erythrocyte membranes. Arminin 1a shows no sequence homology to any known antimicrobial peptide. Because of its high level of activity against multiresistant bacterial strains pathogenic for humans, the peptide arminin 1a is a promising template for a new class of antibiotics. Our data suggest that ancient metazoan organisms such as Hydra hold promise for the detection of novel antimicrobial molecules and the treatment of infections caused by multiresistant bacteria.

Augustin, Rene; Anton-Erxleben, Friederike; Jungnickel, Stephanie; Hemmrich, Georg; Spudy, Bjorn; Podschun, Rainer; Bosch, Thomas C. G.



Activity of the novel peptide arminin against multiresistant human pathogens shows the considerable potential of phylogenetically ancient organisms as drug sources.  


The emergence of multidrug-resistant bacteria highlights the need for new antibacterial agents. Arminin 1a is a novel antimicrobial peptide discovered during investigations of the epithelial defense of the ancient metazoan Hydra. Following proteolytic processing, the 31-amino-acid-long positively charged C-terminal part of arminin 1a exhibits potent and broad-spectrum activity against bacteria, including multiresistant human pathogenic strains, such as methicillin-resistant Staphylococcus aureus (MRSA) strains (minimal bactericidal concentration, 0.4 microM to 0.8 microM). Ultrastructural observations indicate that bacteria are killed by disruption of the bacterial cell wall. Remarkably, the antibacterial activity of arminin 1a is not affected under the physiological salt conditions of human blood. In addition, arminin 1a is a selective antibacterial agent that does not affect human erythrocyte membranes. Arminin 1a shows no sequence homology to any known antimicrobial peptide. Because of its high level of activity against multiresistant bacterial strains pathogenic for humans, the peptide arminin 1a is a promising template for a new class of antibiotics. Our data suggest that ancient metazoan organisms such as Hydra hold promise for the detection of novel antimicrobial molecules and the treatment of infections caused by multiresistant bacteria. PMID:19770277

Augustin, René; Anton-Erxleben, Friederike; Jungnickel, Stephanie; Hemmrich, Georg; Spudy, Björn; Podschun, Rainer; Bosch, Thomas C G



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


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



Intranasal formulation of erythropoietin (EPO) showed potent protective activity against amyloid toxicity in the A?25-35 non-transgenic mouse model of Alzheimer's disease.  


Erythropoietin (EPO) promotes neurogenesis and neuroprotection. We here compared the protection induced by two EPO formulations in a rodent model of Alzheimer's disease (AD): rHu-EPO and a low sialic form, Neuro-EPO. We used the intracerebroventricular administration of aggregated A?25-35 peptide, a non-transgenic AD model. rHu-EPO was tested at 125-500 µg/kg intraperitoneally and Neuro-EPO at 62-250 µg/kg intranasally (IN). Behavioural procedures included spontaneous alternation, passive avoidance, water-maze and object recognition, to address spatial and non-spatial, short- and long-term memories. Biochemical markers of A?25-35 toxicity in the mouse hippocampus were examined and cell loss in the CA1 layer was determined. rHu-EPO and Neuro-EPO led to a significant prevention of A?25-35-induced learning deficits. Both EPO formulations prevented the induction of lipid peroxidation in the hippocampus, showing an antioxidant activity. rHu-EPO (250 µg/kg) or Neuro-EPO (125 µg/kg) prevented the A?25-35-induced increase in Bax level, TNF? and IL-1? production and decrease in Akt activation. A significant prevention of the A?25-35-induced cell loss in CA1 was also observed. EPO is neuroprotective in the A?25-35 AD model, confirming its potential as an endogenous neuroprotection system that could be boosted for therapeutic efficacy. We here identified a new IN formulation of EPO showing high neuroprotective activity. Considering its efficacy, ease and safety, IN Neuro-EPO is a new promising therapeutic agent in AD. PMID:23813967

Maurice, Tangui; Mustafa, Muhammad-Hariri; Desrumaux, Catherine; Keller, Emeline; Naert, Gaëlle; García-Barceló, María de la C; Rodríguez Cruz, Yamila; Garcia Rodríguez, Julío César



Live-cell imaging of endogenous Ras-GTP shows predominant Ras activation at the plasma membrane and in the nucleus in Saccharomyces cerevisiae.  


Ras proteins function as a point of convergence for different signalling pathways in eukaryotes and are involved in many cellular responses; their different subcellular locations could regulate distinct functions. To investigate the localization of active Ras in vivo in Saccharomyces cerevisiae, we expressed a probe consisting of a GFP fusion with a trimeric Ras binding domain of Raf1 (eGFP-RBD3), which binds Ras-GTP with a much higher affinity than Ras-GDP. Our results show that in wild type cells active Ras accumulates mainly at the plasma membrane and in the nucleus during growth on medium containing glucose, while it accumulates mainly in mitochondria in wild type glucose-starved cells and relocalizes to the plasma membrane and to the nucleus upon addition of this sugar. A similar pattern is observed in a strain deleted in the CYR1 gene indicating that the absence of adenylate cyclase does not impair the localization of Ras-GTP. Remarkably, in a gpa2?, but not in a gpr1? mutant, active Ras accumulates in internal membranes and mitochondria, both when cells are growing on glucose medium or are starved, indicating that Gpa2, but not Gpr1 is required for the recruitment of Ras-GTP at the plasma membrane and in the nucleus. Moreover, deletion of both HXK1 and HXK2 also causes a mitochondrial localization of the probe, which relocalizes to the plasma membrane and to the nucleus upon expression of HXK2 on a centromeric plasmid, suggesting that this kinase is involved in the proper localization of active Ras. PMID:23127800

Broggi, Serena; Martegani, Enzo; Colombo, Sonia



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


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



Selective imaging of presynaptic activity in the mouse olfactory bulb shows concentration and structure dependence of odor responses in identified glomeruli  

PubMed Central

More chemicals can be smelled than there are olfactory receptors for them, necessitating a combinatorial representation by somewhat broadly tuned receptors. To understand the perception of odor quality and concentration, it is essential to establish the nature of the receptor repertoires that are activated by particular odorants at particular concentrations. We have taken advantage of the one-to-one correspondence of glomeruli and olfactory receptor molecules in the mouse olfactory bulb to analyze the tuning properties of a major receptor population by high resolution calcium imaging of odor responses selectively in the presynaptic compartment of glomeruli. We show that eighty different olfactory receptors projecting to the dorsal olfactory bulb respond to high concentrations of aldehydes with limited specificity. Varying ensembles of about 10 to 20 receptors encode any particular aldehyde at low stimulus concentrations with high specificity. Even normalized odor response patterns are markedly concentration dependent, caused by pronounced differences in affinity within the aldehyde receptor repertoire.

Fried, Hans U.; Fuss, Stefan H.; Korsching, Sigrun I.



Phthalazine derivatives containing imidazole rings behave as Fe-SOD inhibitors and show remarkable anti-T. cruzi activity in immunodeficient-mouse mode of infection.  


A series of new phthalazine derivatives 1-4 containing imidazole rings were prepared. The monoalkylamino substituted derivatives 2 and 4 were more active in vitro against T. cruzi and less toxic against Vero cells than both their disubstituted analogues and the reference drug benznidazole. Compounds 2 and 4 highly inhibited the antioxidant parasite enzyme Fe-SOD, and molecular modeling suggested that they interact with the H-bonding system of the iron atom moiety. In vivo tests on the acute phase of Chagas disease gave parasitemia inhibition values twice those of benznidazole, and a remarkable decrease in the reactivation of parasitemia was found in the chronic phase for immunodeficient mice. Glucose metabolism studies showed that compounds 1-4 did not affect the succinate pathway but originated important changes in the excretion of pyruvate metabolites. The morphological alterations found in epimastigotes treated with 1-4 confirmed extensive cytoplasm damage and a high mortality rate of parasites. PMID:23043291

Sánchez-Moreno, Manuel; Gómez-Contreras, Fernando; Navarro, Pilar; Marín, Clotilde; Olmo, Francisco; Yunta, María J R; Sanz, Ana María; Rosales, María José; Cano, Carmen; Campayo, Lucrecia



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


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



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


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



In planta analysis of the Agrobacterium tumefaciens T-cyt gene promoter: identification of an upstream region essential for promoter activity in leaf, stem and root cells of transgenic tobacco  

Microsoft Academic Search

The promoter region of the Agrobacterium tumefaciens T-cyt gene was fused to a ß-glucuronidase (gusA) reporter gene and introduced into tobacco plants. Detection of gusA expression in transgenic F1 progeny revealed that the T-cyt promoter is active in many, if not all, cell types in leaves, stems and roots of fully developed plants. Developmental stage-dependent promoter activity was observed in

Saskia T. C. Neuteboom; Esther Hulleman; Rob A. Schilperoort; J. Harry C. Hoge



Th2-like CD8+ T cells showing B cell helper function and reduced cytolytic activity in human immunodeficiency virus type 1 infection  

PubMed Central

We analyzed at clonal level the functional profile of circulating or skin-infiltrating T lymphocytes from two individuals infected with the human immunodeficiency virus type 1 (HIV-1), suffering from a Job's- like syndrome (eczematous dermatitis, recurrent skin and sinopulmonary infections, and hypergammaglobulinemia E) and showing virtually no circulating CD4+ T cells. Most of the CD3+ T cell clones generated from both patients were CD4- CD8+ TCR alpha beta +. The others were CD4- CD8- TCR alpha beta + which exhibited reduced mRNA expression for the CD8 molecule or no mRNA expression for either CD4 or CD8 molecules. The great majority of both CD4- CD8+ and CD4- CD8- did not produce interferon (IFN) gamma and exhibited reduced cytolytic activity. Rather, most of them produced large amounts of both interleukin (IL) 4 and IL-5 and provided B cell helper function for IgE synthesis. These data suggest that a switch of cytolytic CD8+ T cells showing a Th1-like cytokine secretion profile to cells that make Th2-type cytokines, exhibit reduced cytolytic potential, and provide B cell helper function can occur in the course of HIV-1 infection. These cells may contribute to the reduced defense against viral infections and intracellular parasites and account for the elevated IgE serum levels, eosinophilia, and the allergic-like clinical manifestations seen in a proportion of HIV-1-infected individuals.



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


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



The activation domain of transcription factor PU.1 binds the retinoblastoma (RB) protein and the transcription factor TFIID in vitro: RB shows sequence similarity to TFIID and TFIIB.  

PubMed Central

The retinoblastoma (RB) tumor suppressor protein and the TATA-box-binding protein TFIID form contacts with a number of viral transactivator proteins. One of these, the adenovirus E1A protein, can bind to both proteins. Here we present evidence that the cellular transcription factor PU.1 can bind to both RB and TFIID. Like E1A, PU.1 binds to the conserved C-terminal domain of TFIID and to the RB "pocket" domain. The PU.1 sequences required to bind either protein lie within a 75-amino acid region which functions as an independent activation domain in vivo. The ability of PU.1 to contact directly both RB and TFIID through the same 75-residue domain prompted us to look for sequence similarity between these two proteins. We find that the previously defined domain A of the RB pocket shows sequence similarity to the conserved C terminus of TFIID, whereas domain B shows sequence similarity to a second general transcription factor, TFIIB. The potential for RB to influence transcription by using TFIID- and TFIIB-related functions is discussed. Images

Hagemeier, C; Bannister, A J; Cook, A; Kouzarides, T



Peroxidases identified in a subtractive cDNA library approach show tissue-specific transcript abundance and enzyme activity during seed germination of Lepidium sativum.  


The micropylar endosperm is a major regulator of seed germination in endospermic species, to which the close Brassicaceae relatives Arabidopsis thaliana and Lepidium sativum (cress) belong. Cress seeds are about 20 times larger than the seeds of Arabidopsis. This advantage was used to construct a tissue-specific subtractive cDNA library of transcripts that are up-regulated late in the germination process specifically in the micropylar endosperm of cress seeds. The library showed that a number of transcripts known to be up-regulated late during germination are up-regulated in the micropylar endosperm cap. Detailed germination kinetics of SALK lines carrying insertions in genes present in our library showed that the identified transcripts do indeed play roles during germination. Three peroxidases were present in the library. These peroxidases were identified as orthologues of Arabidopsis AtAPX01, AtPrx16, and AtPrxIIE. The corresponding SALK lines displayed significant germination phenotypes. Their transcripts were quantified in specific cress seed tissues during germination in the presence and absence of ABA and they were found to be regulated in a tissue-specific manner. Peroxidase activity, and particularly its regulation by ABA, also differed between radicles and micropylar endosperm caps. Possible implications of this tissue-specificity are discussed. PMID:19884228

Linkies, Ada; Schuster-Sherpa, Uta; Tintelnot, Stefanie; Leubner-Metzger, Gerhard; Müller, Kerstin



Ferredoxin from sweet pepper (Capsicum annuum L.) intensifying harpin(pss)-mediated hypersensitive response shows an enhanced production of active oxygen species (AOS).  


The hypersensitive response (HR) is a form of cell death associated with plant resistance to pathogen infection. Harpin(pss), an elicitor from the bacterium Pseudomonas syringae pv. syringae, induces a HR in non-host plants. Previously, we reported an amphipathic protein from sweet pepper interfering with harpin(pss)-mediated HR. In this report, we isolated and characterized a cDNA clone encoded that amphipathic protein from sweet pepper. This protein is designated as PFLP (plant ferredoxin-like protein) by virtue of its high homology with plant ferredoxin protein containing an N-terminal signal peptide responsible for chloroplast targeting and a putative 2Fe-2S domain responsible for redox activity. Recombinant PFLP obtained from Escherichia coli was able to significantly increase active oxygen species (AOS) generation when mixed with harpin(pss) in tobacco suspension cells. It also showed enhanced HR when co-infiltrated with harpin(pss) in tobacco leaves. We used a transgenic tobacco suspension cells system that constitutively expresses the Pflp gene driven by the CaMV 35S promoter to study the function of PFLP in enhancing harpin(pss)-mediated hypersensitive cell death in vivo. In response to harpin(pss), suspension cells derived from Pflp transgenic tobacco showed a significant increase both in the generation of AOS and in cell death as compared to the wild type. AOS inhibitors diphenylene iodonium chloride (DPI) and lanthanum chlorate (LaCl3) were used to study the involvement of AOS in harpin(pss)-induced cell death. Our results demonstrate enhanced generation of AOS is necessary to cause enhanced hypersensitive cell death in Pflp transgenic tobacco cells and it is plasma membrane-bound NADPH-oxidase-dependent. Sub-cellular localization studies showed that PFLP is present in the cytoplasm and chloroplast of Pflp transgenic tobacco cells, but only in the chloroplast, not in the cytoplasm, of wild-type tobacco cells. It is possible that PFLP can change the redox state of the cell upon harpin(pss) inoculation to increase AOS generation and hypersensitive cell death. Overall, this study will provide a new insight in the functional properties of ferredoxin in hypersensitive cell death. PMID:12777051

Dayakar, Badri Venkata; Lin, Hao-Jan; Chen, Cheng-Hsien; Ger, Mang-Jye; Lee, Bor-Heng; Pai, Chia-Hwei; Chow, David; Huang, Hsiang-En; Hwang, Shaw-Yhi; Chung, Mei-Chu; Feng, Teng-Yung



Field measurements along the 2010 Ms 7.1 Yushu earthquake rupture shows strike-slip and dip-slip activities, resulting in mountains uplift  

NASA Astrophysics Data System (ADS)

The Yushu Ms 7.1 earthquake occurred in the Qinghai Province, China, on April 14th, 2010. Understanding its mechanism is critical to studying the local stress field and the mechanism of earthquake, therefore we conducted careful field investigation immediately after the main shock. Morphological field research shows that the earthquake was triggered by the Ganzi-Yushu fault, trending NW-SE and dipping NE. It spreads at the base of the range-front, along which huge triangular facets (up to 600 m) are distributed, attesting the important vertical component of this fault. Geomorphic features (such as troughs, rivers, fences, and alluvial fans) exhibit sinistral offsets that vary from tens of meters to hundreds of meters. Due to both strike-slip and dip-slip displacements, this fault seems to be a transtensional fault. Thorough observation and measurements were made along the rupture zone, which is about 49 km-long, and consists of 3 discontinuous left-stepping rupture segments (19 km, 22 km, and about 8 km, respectively, from west to east). We observed a maximum sinistral offset of 2.3 m along the central segment and a maximum vertical offset of 0.6 m along the western segment. These offsets, as well as push-up, co-seismic pull-apart and left stepping en-echelon tension fissures, show strike-slip and dip-slip components. The angle (?) between the Principal Displacement Zone (PDZ) and en-echelon tension fissures can reflect the surface rupture kinematics: ? larger than 45 degrees in transpression, ? less than 45 degrees in transtension, and ? equals to 45 degrees in simple shear. For instance, along the Changu Temple segment, we measured 125 rupture directions and found that the mean PDZ strike is ca.295 degrees NW while the fissures’ strike ranges from 278-300 degrees NW. The value of ? is less than 45 degrees, revealing the transtensional regime. In the Guoyangyansongduo segment, we measured 287 rupture directions and found that the PDZ strikes ca.300 degrees NW while fissures strike 265-290 degrees NW, also indicating ? is less than 45 degrees and a transtensional regime. Lastly, in the Longbao Lake segment, 30 rupture direction measurements show that the PDZ strikes 290 degrees NW and fissures strike 270 degrees NW, also showing a transtensional regime. The uplift of the mountain range therefore results from the transtensional regime of the fault and the long term activity resembles Yushu earthquake of this fault.

Fuyao, W.; Li, H.; Pan, J.; Xu, Z.; Li, N.; Guo, R.; Zhang, W.



Characterization of a highly active promoter, PBbgpd, in Beauveria bassiana.  


The promoter of glyceraldehyde-3-phosphate dehydrogenase (gpd) gene from Aspergillus nidulans (PgpdA) is widely used to direct expression of target genes constitutively in fungi. However, in some species, a heterogeneous promoter is found to be of low efficiency. To obtain a high-efficiency promoter for transformation of Beauveria bassiana, an entomopathogenic fungus widely used as an mycoinsecticide, a glyceraldehyde-3-phosphate dehydrogenase gene (Bbgpd) promoter, was cloned and characterized. Four deletion constructs (-2118, -1153, -726, and -354) of the 5'-upstream sequence of Bbgpd linked to a bar::gus fusion gene (phosphinothricin-resistance::beta-glucuronidase fused gene), which were used as selected marker gene and report gene, respectively, were generated. GUS activities of transgenic strains harboring -726, -1153, and -2118 deletion constructs were much stronger than that of the promoter of Aspergillus nidulans gpdA (PgpdA), with a twofold to threefold increase over that in the PgpdA construct. The -726 fragment was necessary to direct GUS expression in B. bassiana. No -354 transgenic progenies were obtained, possibly because it failed to initiate the transcription of bar::gus fusion gene. A remarkable increase of GUS activity was found between the -1153 and -726 constructs, indicating that some active transcriptional elements were located in this region. With a high expression level and relatively short sequence, PBbgpd can be used to drive target genes in B. bassiana transgenic research. PMID:18443858

Liao, Xing-gang; Fang, Wei-guo; Zhang, Yong-jun; Fan, Yan-hua; Wu, Xing-wei; Zhou, Qun; Pei, Yan



Identification of a new mutagenic polychlorinated biphenyl derivative in the Waka River, Wakayama, Japan, showing activation of an aryl hydrocarbon receptor-dependent transcription.  


Water samples from the Waka River, which runs through an area housing many chemical industry facilities in Wakayama, Japan, have been found to show significant mutagenicity, especially without a mammalian metabolic activation system (S9 mix) in the Salmonella typhimurium YG1024 strain. Mutagens in the river water were adsorbed to 3 kg of blue cotton, extracted with methanol/ammonia, and separated by several low- and high-pressure liquid chromatography steps with reversed-phase columns. One mutagen (0.6 mg), accounting for 50% of the total mutagenicity of the adsorbed materials, was isolated. On the basis of the mass, high-resolution mass, (1)H NMR and (13)C NMR spectra, the chemical was determined to have a polychlorinated biphenyl skeleton with nitro and amino substitution groups. Well-designed chemical synthesis of the putative mutagen revealed it to be 4-amino-3,3'-dichloro-5,4'-dinitrobiphenyl. This novel compound exerted strong mutagenicity without the S9 mix, inducing 66,000 and 140,000 revertants/nmol in S. typhimurium TA98 and YG1024, respectively. Moreover, this polychlorinated biphenyl derivative was proven to activate the human aryl hydrocarbon receptor-mediated transcription in a lac Z reporter gene assay with an efficiency almost the same as that of beta-naphthoflavone, well-known to be a synthetic aryl hydrocarbon receptor agonist. It is possible that the mutagen is formed unintentionally via postemission modification of drainage water containing parent chemicals, such as 3,3'-dichlorobenzidine or 3,3'-dichloro-4,4'-dinitrobiphenyl, which are known to be raw materials in the manufacture of polymers and dye intermediates in chemical plants. PMID:11896690

Takamura-Enya, Takeji; Watanabe, Tetsushi; Tada, Atsuko; Hirayama, Teruhisa; Nukaya, Haruo; Sugimura, Takashi; Wakabayashi, Keiji



Healthy working school teachers with high effort-reward-imbalance and overcommitment show increased pro-inflammatory immune activity and a dampened innate immune defence.  


To test whether chronic work stress is accompanied by altered immune functioning, changes in lymphocyte subsets and in lymphocyte production of cytokines were examined in reaction to acute psychosocial stress. Work stress was measured according to Siegrist's effort-reward-imbalance (ERI) model. ERI reflects stress due to a lack of reciprocity between costs and gains at work. Overcommitment (OC) is conceptualized as a dysfunctional coping pattern mainly characterized by the inability to withdraw from work obligations. Fifty-five healthy teachers (34 women, 21 men, mean age 50.0 ± 8.47 years) were exposed to a standardized laboratory stressor (Trier Social Stress Test). Lymphocyte subset counts and lymphocyte production of tumor-necrosis-factor (TNF)-?, interferon (IFN)-?, interleukin (IL)-2, -4, -6 and -10 were measured before and after challenge. High levels of ERI and OC were associated with lower natural killer (NK) cell (CD16+/56+) numbers whereas high levels of OC were related to a lower increase in T-helper cells (CD4+) after stress. Furthermore, subjects with higher ERI showed an overall increased pro-inflammatory activity, with higher TNF-? production at both time points and elevated pre-stress IL-6 production. IL-10 production decreased with higher ERI after stress. The ratios of TNF-?/IL-10 and IL-6/IL-10 were significantly increased in subjects high on ERI. Finally, OC was associated with higher IL-2 production post-stress. The present findings suggest a dampened innate immune defence, reflected in lower NK cell numbers together with an increased pro-inflammatory activity in teachers high on ERI and OC. Such pathways could partly be responsible for the increased vulnerability for stress-related diseases in individuals suffering from chronic work stress. PMID:20599495

Bellingrath, Silja; Rohleder, Nicolas; Kudielka, Brigitte M



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



Antimalarial Iron Chelator, FBS0701, Shows Asexual and Gametocyte Plasmodium falciparum Activity and Single Oral Dose Cure in a Murine Malaria Model  

PubMed Central

Iron chelators for the treatment of malaria have proven therapeutic activity in vitro and in vivo in both humans and mice, but their clinical use is limited by the unsuitable absorption and pharmacokinetic properties of the few available iron chelators. FBS0701, (S)3”-(HO)-desazadesferrithiocin-polyether [DADFT-PE], is an oral iron chelator currently in Phase 2 human studies for the treatment of transfusional iron overload. The drug has very favorable absorption and pharmacokinetic properties allowing for once-daily use to deplete circulating free iron with human plasma concentrations in the high µM range. Here we show that FBS0701 has inhibition concentration 50% (IC50) of 6 µM for Plasmodium falciparum in contrast to the IC50 for deferiprone and deferoxamine at 15 and 30 µM respectively. In combination, FBS0701 interfered with artemisinin parasite inhibition and was additive with chloroquine or quinine parasite inhibition. FBS0701 killed early stage P. falciparum gametocytes. In the P. berghei Thompson suppression test, a single dose of 100 mg/kg reduced day three parasitemia and prolonged survival, but did not cure mice. Treatment with a single oral dose of 100 mg/kg one day after infection with 10 million lethal P. yoelii 17XL cured all the mice. Pretreatment of mice with a single oral dose of FBS0701 seven days or one day before resulted in the cure of some mice. Plasma exposures and other pharmacokinetics parameters in mice of the 100 mg/kg dose are similar to a 3 mg/kg dose in humans. In conclusion, FBS0701 demonstrates a single oral dose cure of the lethal P. yoelii model. Significantly, this effect persists after the chelator has cleared from plasma. FBS0701 was demonstrated to remove labile iron from erythrocytes as well as enter erythrocytes to chelate iron. FBS0701 may find clinically utility as monotherapy, a malarial prophylactic or, more likely, in combination with other antimalarials.

Ferrer, Patricia; Tripathi, Abhai K.; Clark, Martha A.; Hand, Carla Cerami; Rienhoff, Hugh Young; Sullivan, David J.



Analysis of a chimeric class-I patatin-GUS gene in transgenic potato plants: High-level expression in tubers and sucrose-inducible expression in cultured leaf and stem explants  

Microsoft Academic Search

Patatin is a family of lipid acyl hydrolases that accounts for 30 to 40% of the total soluble protein in potato (Solanum tuberosum L.) tubers. To examine the regulation of the patatin genes, we constructed a chimeric gene containing 2.5 kb of 5' flanking sequence from the class I patatin genomic clone PS20 transcriptionally fused to ß-glucuronidase (GUS) and introduced

Herman C. Wenzler; Gregory A. Mignery; Linda M. Fisher; William D. Park



Competition for nodule occupancy on Phaseolus vulgaris by Rhizobium etli and Rhizobium tropici strains can be efficiently monitored in an ultisol during the early stages of growth using a constitutive GUS gene fusion  

Microsoft Academic Search

The GUS gene fusion (pKW107) was transferred into 4 Rhizobium phaseoli strains (KIM5s, CIAT 895, CIAT 7202, CIAT 151) and 1 Rhizobium tropici strain (CIAT 899) in order to determine the nodulation competitiveness of these strains on common bean (Phaseolus vulgaris) in soil. DNA was fingerprinted with the different mutants to verify correct integration of the glucuronidase gene in the

W. Streit; L. Botero; D. Werner; D. Beck



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



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



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.



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



The Great Cometary Show  

NASA Astrophysics Data System (ADS)

The ESO Very Large Telescope Interferometer, which allows astronomers to scrutinise objects with a precision equivalent to that of a 130-m telescope, is proving itself an unequalled success every day. One of the latest instruments installed, AMBER, has led to a flurry of scientific results, an anthology of which is being published this week as special features in the research journal Astronomy & Astrophysics. ESO PR Photo 06a/07 ESO PR Photo 06a/07 The AMBER Instrument "With its unique capabilities, the VLT Interferometer (VLTI) has created itself a niche in which it provide answers to many astronomical questions, from the shape of stars, to discs around stars, to the surroundings of the supermassive black holes in active galaxies," says Jorge Melnick (ESO), the VLT Project Scientist. The VLTI has led to 55 scientific papers already and is in fact producing more than half of the interferometric results worldwide. "With the capability of AMBER to combine up to three of the 8.2-m VLT Unit Telescopes, we can really achieve what nobody else can do," added Fabien Malbet, from the LAOG (France) and the AMBER Project Scientist. Eleven articles will appear this week in Astronomy & Astrophysics' special AMBER section. Three of them describe the unique instrument, while the other eight reveal completely new results about the early and late stages in the life of stars. ESO PR Photo 06b/07 ESO PR Photo 06b/07 The Inner Winds of Eta Carinae The first results presented in this issue cover various fields of stellar and circumstellar physics. Two papers deal with very young solar-like stars, offering new information about the geometry of the surrounding discs and associated outflowing winds. Other articles are devoted to the study of hot active stars of particular interest: Alpha Arae, Kappa Canis Majoris, and CPD -57o2874. They provide new, precise information about their rotating gas envelopes. An important new result concerns the enigmatic object Eta Carinae. Using AMBER with its high spatial and spectral resolution, it was possible to zoom into the very heart of this very massive star. In this innermost region, the observations are dominated by the extremely dense stellar wind that totally obscures the underlying central star. The AMBER observations show that this dense stellar wind is not spherically symmetric, but exhibits a clearly elongated structure. Overall, the AMBER observations confirm that the extremely high mass loss of Eta Carinae's massive central star is non-spherical and much stronger along the poles than in the equatorial plane. This is in agreement with theoretical models that predict such an enhanced polar mass-loss in the case of rapidly rotating stars. ESO PR Photo 06c/07 ESO PR Photo 06c/07 RS Ophiuchi in Outburst Several papers from this special feature focus on the later stages in a star's life. One looks at the binary system Gamma 2 Velorum, which contains the closest example of a star known as a Wolf-Rayet. A single AMBER observation allowed the astronomers to separate the spectra of the two components, offering new insights in the modeling of Wolf-Rayet stars, but made it also possible to measure the separation between the two stars. This led to a new determination of the distance of the system, showing that previous estimates were incorrect. The observations also revealed information on the region where the winds from the two stars collide. The famous binary system RS Ophiuchi, an example of a recurrent nova, was observed just 5 days after it was discovered to be in outburst on 12 February 2006, an event that has been expected for 21 years. AMBER was able to detect the extension of the expanding nova emission. These observations show a complex geometry and kinematics, far from the simple interpretation of a spherical fireball in extension. AMBER has detected a high velocity jet probably perpendicular to the orbital plane of the binary system, and allowed a precise and careful study of the wind and the shockwave coming from the nova. The stream of results from the VLTI and AMBER



Comparative analysis of RNA silencing suppression activities between viral suppressors and an endogenous plant RNA-dependent RNA polymerase.  


RNA silencing is an evolutionarily conserved system that functions as an antiviral mechanism in eukaryotes, including higher plants. To counteract this, several plant viruses express silencing suppressors that inhibit RNA silencing in host plants. Here, we show that both 2b protein from peanut stunt virus (PSV) and a hairpin construct (designated hp-RDR6) that silences endogenous RNA-dependent RNA polymerase 6 (RDR6) strongly suppress RNA silencing. The Agrobacterium infiltration system was used to demonstrate that both PSV 2b and hp-RDR6 suppressed local RNA silencing as strongly as helper component (HC-Pro) from potato virus Y (PVY) and P19 from tomato bush stunt virus (TBSV). The 2b protein from PSV eliminated the small-interfering RNAs (siRNAs) associated with RNA silencing and prevented systemic silencing, similar to 2b protein from cucumber mosaic virus (CMV). On the other hand, hp-RDR6 suppressed RNA silencing by inhibiting the generation of secondary siRNAs. The small coat protein (SCP) of squash mosaic virus (SqMV) also displayed weak suppression activity of RNA silencing. Agrobacterium-mediated gene transfer was used to investigate whether viral silencing suppressors or hp-RDR6 enhanced accumulations of green fluorescence protein (GFP) and ?-glucuronidase (GUS) as markers of expression in leaf tissues of Nicotina benthamiana. Expression of both GFP and GUS was significantly enhanced in the presence of PSV 2b or CMV 2b, compared to no suppression or the weak SqMV SCP suppressor. Co-expression with hp-RDR6 also significantly increased the expression of GFP and GUS to levels similar to those induced by PVY HC-Pro and TBSV P19. PMID:22354861

Yoon, Ju-Yeon; Han, Kyoung-Sik; Park, Han-Yong; Choi, Seung-Kook



Endosperm-specific activity of a storage protein gene promoter in transgenic wheat seed.  


The characterization of the promoter of a wheat (Triticum aestivum) cv. Cheyenne high molecular weight glutenin subunit (HMW subunit) gene, Glu-1D-1 is reported. The nucleotide sequence of the promoter from position -1191 to -650 with respect to the transcription start site was determined, to add to that already determined. Analysis of this region of the promoter revealed the presence of an additional copy of part of the primary enhancer sequence and sequences related to regulatory elements present in other wheat seed protein genes. A chimaeric gene was constructed comprising the 5' flanking region of the Glu-1D-1 gene from position -1191 to +58, the coding region of the UID:A (Gus) gene, and the nopaline synthase (Nos) gene terminator. This chimaeric gene was introduced into wheat (Triticum durum cv. Ofanto) by particle bombardment of inflorescence explants. Two independent transgenic lines were produced, and both showed expression of the Gus gene specifically in the endosperm during mid-development (first detected 10-12 d after anthesis). Histochemical analysis of homozygous T(2) seed confirmed this pattern of expression, and showed that expression was initiated first in the central lobes of the starchy endosperm, and then spread throughout the endosperm tissue, while no expression was detected in the aleurone layer. Native HMW subunit protein was detectable by Western analysis 12-14 d after anthesis, consistent with concurrent onset of activity of the native and introduced HMW subunit gene promoters. PMID:11283168

Lamacchia, C; Shewry, P R; Di Fonzo, N; Forsyth, J L; Harris, N; Lazzeri, P A; Napier, J A; Halford, N G; Barcelo, P



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


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 beta-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 antifungal substance p-coumaric acid. Transcriptional up-regulation of MAGGY RNA was also observed following heat shock and oxidative stress. The MAGGY promoter remained responsive to the above-mentioned stresses when transformed into a M. grisea isolate that had no endogenous MAGGY elements. In C. lagenarium, however, the MAGGY promoter showed only basal expression of GUS and no further up-regulation was induced by any of the stress treatments, suggesting that the stress-responding cis-element(s) in the MAGGY promoter is not functional in a wider range of fungi. The relationship between the activation of MAGGY by stress and phenotypic diversification in M. grisea, including variations in pathogenicity, is discussed. PMID:11683275

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



Positive Control Mutations in the MyoD Basic Region Fail to Show Cooperative DNA Binding and Transcriptional Activation in vitro  

NASA Astrophysics Data System (ADS)

An in vitro transcription system from HeLa cells has been established in which MyoD and E47 proteins activate transcription both as homodimers and heterodimers. However, heterodimers activate transcription more efficiently than homodimers, and function synergistically from multiple binding sites. Positive control mutants in the basic region of MyoD that have previously been shown to be defective in initiating the myogenic program, can bind DNA but have lost their ability to function as transcriptional activators in vitro. Additionally, positive control mutants, unlike wild-type MyoD, fail to bind cooperatively to DNA. We propose that binding of MyoD complexes to high affinity MyoD binding sites induces conformational changes that facilitate cooperative binding to multiple sites and promote transcriptional activation.

Bengal, Eyal; Flores, Osvaldo; Rangarajan, Pundi N.; Chen, Amy; Weintraub, Harold; Verma, Inder M.



Mitotic activity of Sertoli cells in adult human testis: an immunohistochemical study to characterize Sertoli cells in testicular cords from patients showing testicular dysgenesis syndrome  

Microsoft Academic Search

During puberty, normal somatic Sertoli cells undergo dramatic morphological changes due to the differentiation of immature\\u000a pre-Sertoli cells in functionally active adult Sertoli cells. Sertoli cell maturation is accompanied with loss of their mitotic\\u000a activity before onset of spermatogenesis and loss of pre-pubertal and occurrence of adult immunohistochemical Sertoli cell\\u000a differentiation markers. Testes of infertile adult patients often exhibit numerous

Ralph Brehm; Rodolfo Rey; Sabine Kliesch; Klaus Steger; Alexander Marks; Martin Bergmann



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


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 PrfA(allo) 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. PMID:22646689

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



Slide Show Template  

Center for Biologics Evaluation and Research (CBER)

Text Version... Basic protocol approach: – Search Strategy for identifying TEE: Medical Dictionary for Regulatory Activities ... After IVIG Infusion Unknown All 1 (1%) ... More results from


DP7, a novel dihydropyridine multidrug resistance reverter, shows only weak inhibitory activity on human CYP3A enzyme(s)  

Microsoft Academic Search

The aim of this study was to investigate the effects of 3,5-dibenzoyl-4-(3-phenoxy-phenyl)-1,4-dihydro-2,6-dimethylpyridine (DP7), a novel multidrug resistance (MDR) reverter, on cytochrome P450 (CYP)-activities by human and rat liver microsomes. Effects of DP7 were assessed with use of selective substrates, markers of CYP activities. With rat microsomes, ethoxyresorufin (ETR) was used as substrate for CYP1A1, penthoxyresorufin (PTR) for 2B, benzyloxyresorufin (BZR)

Paolo D'Elia; Francesco De Matteis; Stefania Dragoni; Anamik Shah; Giampietro Sgaragli; Massimo Valoti



Oscillatory Activity in Human Parietal and Occipital Cortex Shows Hemispheric Lateralization and Memory Effects in a Delayed Double-Step Saccade Task  

Microsoft Academic Search

We applied magnetoencephalography (MEG) to record oscillatory brain activity from human subjects engaged in planning a double- step saccade. In the experiments, subjects (n 5 8) remembered the locations of 2 sequentially flashed targets (each followed by a 2-s delay), presented in either the left or right visual hemifield, and then made saccades to the 2 locations in sequence. We

W. Pieter Medendorp; Geerten F. I. Kramer; Ole Jensen; Robert Oostenveld; Jan-Mathijs Schoffelen; P. Fries



Neurons in the Globus Pallidus Do Not Show Correlated Activity in the Normal Monkey, but Phase-Locked Oscillations Appear in the MPTP Model of Pxkinsonism  

Microsoft Academic Search

1. To test the mode of functional connectivity in the basal gan- glia circuitry, we studied the activity of simultaneously recorded neurons in the globus pallidus (GP) of a behaving rhesus monkey. The cross-correlograms of pairs of neurons in the GP were com- pared with those of neurons in the thalamus and frontal cortex and to the cross-correlograms of pallidal



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



The Diane Rehm Show  

NSDL National Science Digital Library

The Diane Rehm Show has its origins in a mid-day program at WAMU in Washington, D.C. Diane Rehm came on to host the program in 1979, and in 1984 it was renamed "The Diane Rehm Show". Over the past several decades, Rehm has played host to hundreds of guests, include Archbishop Desmond Tutu, Julie Andrews, and President Bill Clinton. This website contains an archive of her past programs, and visitors can use the interactive calendar to look through past shows. Those visitors looking for specific topics can use the "Topics" list on the left-hand side of the page, or also take advantage of the search engine. The show has a number of social networking links, including a Facebook page and a Twitter feed.


Variant histone H3.3 is deposited at sites of nucleosomal displacement throughout transcribed genes while active histone modifications show a promoter-proximal bias  

PubMed Central

Deposition of variant histones provides a mechanism to reset and to potentially specify chromatin states. We determined the distribution of H3 and its variant H3.3 relative to chromatin structure and elongating polymerase. H3.3 is enriched throughout active genes similar to polymerase, yet its distribution is very distinct from that of several euchromatic histone modifications, which are highly biased toward the 5? part of active genes. Upon gene induction we observe displacement of both H3 and H3.3 followed by selective deposition of H3.3. These results support a model in which H3.3 deposition compensates for transcription-coupled nucleosomal displacement yet does not predetermine tail modifications.

Wirbelauer, Christiane; Bell, Oliver; Schubeler, Dirk



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

Microsoft Academic Search

Induction of plant-derived chitinases in the leaves of a carnivorous plant was demonstrated using aseptically grown round-leaf\\u000a sundew (Drosera rotundifolia L.). The presence of insect prey was mimicked by placing the chemical inducers gelatine, salicylic acid and crustacean chitin\\u000a on leaves. In addition, mechanical stirring of tentacles was performed. Chitinase activity was markedly increased in leaf\\u000a exudates upon application of

Ildikó Matušíková; Ján Salaj; Jana Morav?íková; Ludmila Mlynárová; Jan-Peter Nap; Jana Libantová



Adenoviruses with Tcf binding sites in multiple early promoters show enhanced selectivity for tumour cells with constitutive activation of the wnt signalling pathway  

Microsoft Academic Search

Mutation of the adenomatous polyposis coli and ?-catenin genes in colon cancer leads to constitutive activation of transcription from promoters containing binding sites for Tcf\\/LEF transcription factors. We have constructed adenoviruses with Tcf binding sites in the early promoters, in order to target viral replication to colon tumours. Tcf regulation of the E1A promoter confers a 100-fold selectivity for cells

C Fuerer; R Iggo



An Activation Domain of the Helix-Loop-Helix Transcription Factor E2A Shows Cell Type Preference In Vivo in Microinjected Zebra Fish Embryos  

Microsoft Academic Search

The E2A protein is a mammalian transcription factor of the helix-loop-helix family which is implicated in cell-specific gene expression in several cell lineages. Mouse E2A contains two independent transcription activation domains, ADI and ADII; whereas ADI functions effectively in a variety of cultured cell lines, ADII showspreferentialactivityinpancreaticbetacells.Toanalyzethispreferentialactivityinaninvivosetting,we adapted a system involving transient gene expression in microinjected zebra fish embryos. Fertilized




Chromosome 13q12 encoded Rho GTPase activating protein suppresses growth of breast carcinoma cells, and yeast two-hybrid screen shows its interaction with several proteins  

Microsoft Academic Search

We have characterized the cDNA for a Rho GTPase activating protein (GAP) mapping to chromosome 13q12. The cDNA was characterized by determining the complete sequence of a 4.8kb cDNA clone that represents the 5? untranslated region (UTR), the translated region, and the 3? UTR. The protein has a sterile ?-motif (SAM), a distinct GAP domain, and a conserved START (StAR

Ganachari M Nagaraja; Raj P Kandpal



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

Microsoft Academic Search

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

M. Penza; M. Jeremic; E. Marrazzo; A. Maggi; P. Ciana; G. Rando; P. G. Grigolato; D. Di Lorenzo



Field measurements along the 2010 Ms 7.1 Yushu earthquake rupture shows strike-slip and dip-slip activities, resulting in mountains uplift  

Microsoft Academic Search

The Yushu Ms 7.1 earthquake occurred in the Qinghai Province, China, on April 14th, 2010. Understanding its mechanism is critical to studying the local stress field and the mechanism of earthquake, therefore we conducted careful field investigation immediately after the main shock. Morphological field research shows that the earthquake was triggered by the Ganzi-Yushu fault, trending NW-SE and dipping NE.

W. Fuyao; H. Li; J. Pan; Z. Xu; N. Li; R. Guo; W. Zhang



Do Elephants Show Empathy?  

Microsoft Academic Search

Elephants show a rich social organization and display a number of unusual traits. In this paper, we analyse reports collected over a thirty-five year period, describing behaviour that has the potential to reveal signs of empathic understanding. These include coalition formation, the offering of protection and comfort to others, retrieving and 'babysitting' calves, aiding individuals that would otherwise have difficulty

Lucy A. Bates; Phyllis C. Lee; Norah Njiraini; Joyce H. Poole; Katito Sayialel; Soila Sayialel; Cynthia J. Moss; Richard W. Byrne



Shakespearean Slide Shows.  

ERIC Educational Resources Information Center

|Presents a condensed method for involving students in the kind of theatrical problem-solving that transforms a script to a play. Describes how to incorporate a "human slide show" into the class. Notes that students must read plays not just to understand events, but to make artistic choices about how to stage the action so that an audience…

Flynn, Rosalind M.



Btu accounting: Showing results  

Microsoft Academic Search

In the preceding article in this series last month, the author showed how to calculate the energy consumed to make a pound of product. To realize a payoff, however, the results must be presented in graphs or tables that clearly display what has happened. They must call attention to plant performance and ultimately lead to more efficient use of energy.



Stage a Water Show  

ERIC Educational Resources Information Center

In the author's book titled "The Incredible Water Show," the characters from "Miss Alaineus: A Vocabulary Disaster" used an ocean of information to stage an inventive performance about the water cycle. In this article, the author relates how she turned the story into hands-on science teaching for real-life fifth-grade students. The author also…

Frasier, Debra



The Truman Show  

Microsoft Academic Search

The Truman Show is hardly a film you would automatically speak about as a game. At first glance, it is tempting to interpret the story of\\u000a Truman Burbank — his perpetual subjection to the artificial (televisual) world of Seahaven and its gargantuan reality TV project,\\u000a his eventual escape from the “OmniCam Ecosphere” building and the paternalistic surveillance of director Christof

Rolf F. Nohr


Cloning of a G-protein-coupled receptor that shows an activity to transform NIH3T3 cells and is expressed in gastric cancer cells.  


The present study was directed towards the identification of novel factors involved in the transformation process leading to the formation of gastric cancer. A cDNA library from human gastric cancer cells was constructed using a retroviral vector. Functional cloning was performed by screening for transformation activity in transduced NIH3T3 cells. Six cDNA clones were isolated, including one encoding the elongation factor 1alpha subunit, which was already known to play a role in tumorigenesis. One cDNA (clone 56.2), which was repeatedly isolated during the course of screening, encoded a protein identical to a G-protein-coupled receptor protein, GPR35. In addition, another cDNA clone (72.3) was found to be an alternatively spliced product of the GPR35 gene, whereby 31 amino acids were added to the N-terminus of GPR35. Hence, the proteins encoded by clones 56.2 and 72.3 were designated GPR35a and GPR35b, respectively. RT-PCR experiments revealed that GPR35 gene expression is low or absent in surrounding non-cancerous regions, while both mRNAs were present in all of the gastric cancers examined. The level of 72.3-encoded mRNA was consistently significantly higher than that of 56.2 encoded mRNA. An expression pattern similar to that observed in gastric cancers was detected in normal intestinal mucosa. Based on the apparent transformation activities of the two GPR35 clones in NIH3T3 cells, and the marked up-regulation of their expression levels in cancer tissues, it is speculated that these two novel isoforms of GPR35 are involved in the course of gastric cancer formation. PMID:14965362

Okumura, Shun-ichiro; Baba, Hiroko; Kumada, Tatsuro; Nanmoku, Koji; Nakajima, Hirofumi; Nakane, Yasushi; Hioki, Koshiro; Ikenaka, Kazuhiro



Laninamivir Prodrug CS-8958, a Long-Acting Neuraminidase Inhibitor, Shows Superior Anti-Influenza Virus Activity after a Single Administration?  

PubMed Central

Two neuraminidase (NA) inhibitors, zanamivir (Relenza) and oseltamivir phosphate (Tamiflu), have been licensed for use for the treatment and prophylaxis of influenza. We have reported on laninamivir (code name, R-125489), a novel neuraminidase inhibitor, and have discovered that the laninamivir prodrug CS-8958 worked as a long-acting neuraminidase inhibitor in a mouse influenza virus infection model when it is intranasally administered. In this study, CS-8958 was administered just once 7 days before infection and showed significant efficacy in vivo. The efficacy of a single administration of CS-8958 after viral infection was then compared with that of repeated administrations of oseltamivir phosphate or zanamivir in mice and ferrets. CS-8958 showed efficacy superior or similar to the efficacies of the two licensed NA inhibitors. CS-8958 also significantly reduced the titers of an oseltamivir-resistant H1N1 virus with a neuraminidase H274Y substitution in a mouse infection model. These results suggest that since CS-8958 is characteristically long lasting in the lungs, it may be ideal for the prophylaxis and treatment of influenza.

Kubo, Shuku; Tomozawa, Takanori; Kakuta, Masayo; Tokumitsu, Akane; Yamashita, Makoto



Mouse gastric tumor models with prostaglandin E2 pathway activation show similar gene expression profiles to intestinal-type human gastric cancer  

PubMed Central

Background Gastric cancers are generally classified into better differentiated intestinal-type tumor and poorly differentiated diffuse-type one according to Lauren's histological categorization. Although induction of prostaglandin E2 pathway promotes gastric tumors in mice in cooperation with deregulated Wnt or BMP signalings, it has remained unresolved whether the gastric tumor mouse models recapitulate either of human gastric cancer type. This study assessed the similarity in expression profiling between gastric tumors of transgenic mice and various tissues of human cancers to find best-fit human tumors for the transgenic mice models. Results Global expression profiling initially found gastric tumors from COX-2/mPGES-1 (C2mE)-related transgenic mice (K19-C2mE, K19-Wnt1/C2mE, and K19-Nog/C2mE) resembled gastric cancers among the several tissues of human cancers including colon, breast, lung and gastric tumors. Next, classification of the C2mE-related transgenic mice by a gene signature to distinguish human intestinal- and diffuse-type tumors showed C2mE-related transgenic mice were more similar to intestinal-type compared with diffuse one. We finally revealed that induction of Wnt pathway cooperating with the prostaglandin E2 pathway in mice (K19-Wnt1/C2mE mice) further reproduce features of human gastric intestinal-type tumors. Conclusion We demonstrated that C2mE-related transgenic mice show significant similarity to intestinal-type gastric cancer when analyzed by global expression profiling. These results suggest that the C2mE-related transgenic mice, especially K19-Wnt1/C2mE mice, serve as a best-fit model to study molecular mechanism underlying the tumorigenesis of human gastric intestinal-type cancers.



Polytene chromosomes show normal gene activity but some mRNAs are abnormally accumulated in the pseudonurse cell nuclei of Drosophila melanogaster otu mutants.  


Certain mutant alleles of the ovarian tumor (otu) locus give rise to polytene chromosomes in the pseudonurse cells (PNCs). We have previously shown that the banding pattern of these germ line-derived chromosomes is similar to that in the larval salivary gland chromosomes. In this study, we have examined the gene activity of these chromosomes. General gene expression from these chromosomes was studied by uridine autoradiography. The expression of specific genes was monitored by in situ hybridisation to mRNA and also by combining enhancer trap lines with otu mutants. We found that most of the genes studied were expressed in the PNCs as they were in the wild-type nurse cells. Four out of the 12 mRNAs studied accumulated in the nuclei instead of migrating to the cytoplasm. The intensity of accumulation directly correlated with the extent of polytenisation in the PNC nuclei. We suggest that the otu mRNA remains partly attached to the polytene chromosome template after transcription and discuss the effects of this phenomenon on polytenisation of the PNC chromosomes. PMID:7587594

Heino, T I; Lahti, V P; Tirronen, M; Roos, C



A new case of Carney triad: gastrointestinal stromal tumours and leiomyoma of the oesophagus do not show activating mutations of KIT and platelet-derived growth factor receptor ?  

PubMed Central

The Carney triad is a rare syndrome of unknown aetiology, with synchronous or metachronous appearance of rare neoplasms: gastrointestinal stromal tumours (GISTs), pulmonary chondromas and extra?adrenal paragangliomas. In most cases, the Carney triad is incomplete. The combination encountered typically, GISTs and pulmonary chondromas, was also seen in our patient, a 22?year?old woman. She was diagnosed with the triad after Billroth II gastrectomy for histologically proved gastric GISTs. The diagnosis of pulmonary chondromas was confirmed by transthoracic, computed tomography?guided needle biopsy. An oesophageal leiomyoma was resected 2?years after the initial diagnosis, on suspicion of paraganglioma. The clinical course of the patient has been uneventful since. The last follow?up was carried out 6?years after the initial diagnosis. On histological examination, the cells of gastric GIST were partly positive for CD34, whereas CD117 was expressed in all areas in variable intensity and S?100 protein was negative. The oesophageal tumour was classified as leiomyoma due to strong immunopositivity for smooth muscle actin and desmin, being negative for CD34 and CD117. Two different gastric GIST lesions as well as the oesophageal leiomyoma and normal tissue were analysed for activating mutations in common hot spots of KIT (exon 9 and 11) and platelet?derived growth factor receptor ? (exon 18), but in all probes wild?type sequences were found. These results are in accordance with the first published analyses of GIST lesions from Carney patients.

Knop, S; Schupp, M; Wardelmann, E; Stueker, D; Horger, M S; Kanz, L; Einsele, H; Kroeber, S M



Heterogeneous T-cell response to MAGE-A10(254-262): high avidity-specific cytolytic T lymphocytes show superior antitumor activity.  


MAGE-encoded antigens, which are expressed by tumors of many histological types but not in normal tissues, are suitable candidates for vaccine-based immunotherapy of cancers. Thus far, however, T-cell responses to MAGE antigens have been detected only occasionally in cancer patients. In contrast, by using HLA/peptide fluorescent tetramers, we have observed recently that CD8(+) T cells specific for peptide MAGE-A10(254-262) can be detected frequently in peptide-stimulated peripheral blood mononuclear cells from HLA-A2-expressing melanoma patients and healthy donors. On the basis of these results, antitumoral vaccination trials using peptide MAGE-A10(254-262) have been implemented recently. In the present study, we have characterized MAGE-A10(254-262)-specific CD8(+) T cells in polyclonal cultures and at the clonal level. The results indicate that the repertoire of MAGE-A10(254-262)-specific CD8(+) T cells is diverse both in terms of clonal composition, efficiency of peptide recognition, and tumor-specific lytic activity. Importantly, only CD8(+) T cells able to recognize the antigenic peptide with high efficiency are able to lyse MAGE-A10-expressing tumor cells. Under defined experimental conditions, the tetramer staining intensity exhibited by MAGE-A10(254-262)-specific CD8(+) T cells correlates with efficiency of peptide recognition so that "high" and "low" avidity cells can be separated by FACS. Altogether, the data reported here provide evidence for functional diversity of MAGE-A10(254-262)-specific T cells and will be instrumental for the monitoring of peptide MAGE-A10(254-262)-based clinical trials. PMID:11479225

Dutoit, V; Rubio-Godoy, V; Dietrich, P Y; Quiqueres, A L; Schnuriger, V; Rimoldi, D; Liénard, D; Speiser, D; Guillaume, P; Batard, P; Cerottini, J C; Romero, P; Valmori, D



TAK-960, a novel, orally available, selective inhibitor of polo-like kinase 1, shows broad-spectrum preclinical antitumor activity in multiple dosing regimens.  


Polo-like kinase 1 (PLK1) is a serine/threonine protein kinase involved in key processes during mitosis. Human PLK1 has been shown to be overexpressed in various human cancers, and elevated levels of PLK1 have been associated with poor prognosis, making it an attractive target for anticancer therapy. TAK-960 [4-[(9-cyclopentyl-7,7-difluoro-5-methyl-6-oxo-6,7,8,9-tetrahydro-5H-pyrimido[4,5-b][1,4]diazepin-2-yl)amino]-2-fluoro-5-methoxy-N-(1-methylpiperidin-4-yl) benzamide] is a novel, investigational, orally bioavailable, potent, and selective PLK1 inhibitor that has shown activity in several tumor cell lines, including those that express multidrug-resistant protein 1 (MDR1). Consistent with PLK1 inhibition, TAK-960 treatment caused accumulation of G(2)-M cells, aberrant polo mitosis morphology, and increased phosphorylation of histone H3 (pHH3) in vitro and in vivo. TAK-960 inhibited proliferation of multiple cancer cell lines, with mean EC(50) values ranging from 8.4 to 46.9 nmol/L, but not in nondividing normal cells (EC(50) >1,000 nmol/L). The mutation status of TP53 or KRAS and MDR1 expression did not correlate with the potency of TAK-960 in the cell lines tested. In animal models, oral administration of TAK-960 increased pHH3 in a dose-dependent manner and significantly inhibited the growth of HT-29 colorectal cancer xenografts. Treatment with once daily TAK-960 exhibited significant efficacy against multiple tumor xenografts, including an adriamycin/paclitaxel-resistant xenograft model and a disseminated leukemia model. TAK-960 has entered clinical evaluation in patients with advanced cancers. PMID:22188812

Hikichi, Yuichi; Honda, Kohei; Hikami, Kouki; Miyashita, Hitoshi; Kaieda, Isao; Murai, Saomi; Uchiyama, Noriko; Hasegawa, Maki; Kawamoto, Tomohiro; Sato, Takashi; Ichikawa, Takashi; Cao, Sheldon; Nie, Zhe; Zhang, Lilly; Yang, Johnny; Kuida, Keisuke; Kupperman, Erik



NPR: The Picture Show  

NSDL National Science Digital Library

National Public Radio's "The Picture Show" photo blog is a great way to avoid culling through the thousands of less interesting and engaging photographs on the web. With a dedicated team of professionals, this blog brings together different posts that profile various sets of photographs that cover 19th century war in Afghanistan, visual memories of WWII, unpublished photographs of JFK's presidential campaign, and abandoned buildings on the islands in Boston Harbor. Visitors can search through previous posts, use social media features to share the photo features with friends, and also sign up to receive new materials via their RSS feed. There's quite a nice mix of material here, and visitors can also comment on the photos and recommend the collection to friends and others.


Egg: the Arts Show  

NSDL National Science Digital Library

"Egg is a new TV show about people making art across America" from PBS. This accompanying Website presents excerpts from sixteen episodes of the series, with three more "hatching soon," such as Close to Home, profiling three photographers: Jeanine Pohlhaus, whose pictures document her father's struggle with mental illness; Gregory Crewdson's photos of Lee, Massachusetts; and Joseph Rodriguez's photos of Hispanics in New York City. Excerpts include video clips, gallery listings where the artists' work can be seen, and short interviews with artists. Some episodes also offer "peeps," glimpses of material not shown on TV, such as the Space episode's peep, Shooting Stars, that provides directions for astrophotography, taking photographs of star trails. Other sections of the site are airdates, for local listings; see and do usa, where vacationers can search for art events at their destinations; and egg on the arts, a discussion forum.


American History Picture Show  

NSDL National Science Digital Library

In class we read Katie's Picture Show, a book about a girl who discovers art first-hand one day at an art museum in London. She realizes she can climb into the paintings, explore her surroundings, and even solve problems for the subjects of the paintings. As part of our unit on American history, we are going to use art to further learn about some of the important events we have been discussing. Each of these works of art depicts an important event in American History. When you click on a picture, you will be able to see the name of the event as well as the artist who created it. You will be using all three pictures for this assignment.Use the websites ...

Bennion, Ms.



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.



A rapamycin-binding protein polymer nanoparticle shows potent therapeutic activity in suppressing autoimmune dacryoadenitis in a mouse model of Sjögren's syndrome.  


Sjögren's syndrome (SjS) is a chronic autoimmune disease characterized initially by lymphocytic infiltration and destruction of exocrine glands, followed by systemic organ damage and B-cell lymphoma. Conventional treatment is based on management of symptoms and there is a shortage of therapies that address the underlying causes of inflammation at source exocrine tissue. The aim of this study was to test a novel protein polymer-based platform consisting of diblock copolymers composed from Elastin-like Polypeptides (ELPs) fused with FKBP12, to deliver a potent immunosuppressant with dose-limiting toxicity, rapamycin (Rapa) also known as Sirolimus, and evaluate its effects on the inflamed lacrimal gland (LG) of non-obese diabetic mouse (NOD), a classic mouse model of SjS. Both soluble and diblock copolymer ELPs were fused to FKBP12 and characterized with respect to purity, hydrodynamic radii, drug entrapment and release. Both formulations showed successful association with Rapa; however, the nanoparticle formulation, FSI, released drug with nearly a 5 fold longer terminal half-life of 62.5h. The strong interaction of FSI nanoparticles with Rapa was confirmed in vivo by a shift in the monoexponential pharmacokinetic profile for free drug to a biexponential profile for the nanoparticle formulation. When acutely administered by injection into NOD mice via the tail vein, this FSI formulation significantly suppressed lymphocytic infiltration in the LG relative to the control group while reducing toxicity. There was also a significant effect on inflammatory and mammalian target of Rapamycin (mTOR) pathway genes in the LG and surprisingly, our nanoparticle formulation was significantly better at decreasing a proposed tear biomarker of SjS, cathepsin S (CATS) compared to free drug. These findings suggest that FSI is a promising tool for delivering Rapa for treatment of SjS in a murine model and may be further explored to meet the unmet medical challenge of SjS. PMID:23892265

Shah, Mihir; Edman, Maria C; Janga, Srikanth R; Shi, Pu; Dhandhukia, Jugal; Liu, Siyu; Louie, Stan G; Rodgers, Kathleen; Mackay, J Andrew; Hamm-Alvarez, Sarah F



Cassini UVIS Observations Show Active Saturn's Rings  

NASA Astrophysics Data System (ADS)

The Cassini Ultraviolet Imaging Spectrograph (UVIS) is part of the remote sensing payload of the NASA/ESA Cassini spacecraft. This spectrograph includes channels for extreme UV and far UV spectroscopic imaging, high speed photometry of stellar occultations, solar EUV occultation, and a hydrogen/deuterium absorption cell. We report our initial results from UVIS observations of Saturn's rings. Dynamic interactions between neutrals, ions, rings, moons and meteoroids produce a highly structured and time variable Saturn system Oxygen in the Saturn system dominates the magnetosphere. Observed fluctuations indicate close interactions with plasma sources. Stochastic events in the E ring may be the ultimate source. The spectral signature of water ice is seen on Phoebe and in Saturn's rings. Water ice is mixed non-uniformly with darker constituents. The high structure of the UV ring reflectance argues that collisional transport dominates ballistic transport in darkening the rings. Our preliminary results support the idea that rings are recycled fragments of moons: the current processes are more important than history and initial conditions. The spectra along the UVIS SOI radial scan indicate varying amounts of water ice. In the A ring, the ice fraction increases outward to a maximum at the outer edge. This large-scale variation is consistent with initially pure ice that has suffered meteoritic bombardment over the age of the Solar system (Cuzzi and Estrada 1998). We also see variations over scales of 1000 - 3000 km, which cannot be explained by this mechanism. Ballistic transport of spectrally neutral extrinsic pollutants from meteoroids striking the rings has a typical throw distance of 6000 km (Durisen et al 1989), too long to explain this finer structure. We propose a class of smaller renewal events, in which a small moon residing within the rings is shattered by an external impactor (Colwell and Esposito 1993, Barbara and Esposito 2002, Esposito and Colwell 2003). The interior of such a body has been shielded from external meteoritic bombardment, and thus contains purer ice. Since the amount of meteoroid pollution provides a rough clock to estimate the age of the rings (Cuzzi and Estrada), these random events reset that clock locally, making the material at that radial location younger and purer. As these purer ring particles collide with others, they exchange regolith, and the range of purer water ice spectrum spreads radially. The radial variation we interpret as due to differential pollution in our data set is consistent with the disruption of several small bodies in the A ring in the last 107 to 108 years. When the small moon Pan (Showalter 1991, R ~ 10 km, now residing in the nearby Encke Gap) is eventually shattered by an external impact (Colwell et al 2000), the gap will close up, and for some 10 to 100 million years thereafter a brighter radial swath of purer water ice at its former location will gradually spread and darken.

Esposito, L.; Colwell, J. E.; UVIS Team



Isolation and promoter characterization of barley gene Itr1 encoding trypsin inhibitor BTI-CMe: differential activity in wild-type and mutant lys3a endosperm.  


The gene Itr1, encoding trypsin inhibitor BTI-CMe, has been obtained from a genomic library of Hordeum vulgare L. The gene has no introns and presents in its 5'-upstream region 605 bp that are homologous to the long terminal repeats (LTR) of the 'copia-like' retro-transposon Bare-1. Functional analysis of the Itr1 promoter by transient expression in protoplasts derived from different barley tissues, has shown that in this system the Itr1 promoter retains its endosperm specifity and the trans-regulation mediated by the Lys3a gene. The proximal promoter extending 343 bp upstream of the translation initiation ATG codon is sufficient to confer full GUS expression and for endosperm specifity. In protoplasts derived from the lys3a mutant, Risř 1508, GUS activity was less than 5% of that obtained with the same constructs in the protoplasts of wild-type Bomi from which it derives. Gel retardation experiments, after incubation with proteins obtained from both types of endosperm nuclei, also show differential patterns. Possible reasons for these differences are discussed. PMID:8843947

Royo, J; Diaz, I; Rodriquez-Palenzuela, P; Carbonero, P



Generative cell-specific activation of the histone gH2A gene promoter of Lilium longiflorum in tobacco.  


The Lilium longiflorum gH2A promoter is active exclusively in the generative cells of mature pollen in transgenic tobacco expressing the gH2A promoter::GUS (?-glucuronidase) construct as a reporter gene. Temporal and spatial aspects of gH2A promoter activity examined during pollen development in transgenic tobacco reveal that GUS reporter activity was not detected until developing pollen entered the early bicellular developmental stage. Activity was first detected in generative cells at early-mid stages and gradually increased to maximum levels at mid-bicellular stages. The patterns of appearance and longevity of GUS activity in tobacco were very similar to those of gH2A mRNA during pollen development in Lilium. Exogenous treatment with colchicine, a well-known microtubule depolymerize, blocked microspore mitosis and inhibited generative cell differentiation. No GUS signal was detected in the resulting anomalous pollen, which lacked generative cell differentiation. These data strongly suggest that normal generative cell development is essential for activation of the gH2A promoter. Furthermore, these results indicate that common transcriptional activator(s) of the gH2A promoter may be present in both Lilium and Nicotiana, and that such putative factor(s) activates the gH2A promoter only when generative cells undergo normal development. PMID:22820801

Ueda, Kenji; Ono, Michiyuki; Iwashita, Jun; Wabiko, Hiroetsu; Inoue, Masayasu



The expression of mRNAs for hepatocyte growth factor\\/scatter factor, its receptor c- met, and one of its activators tissue-type plasminogen activator show a systematic relationship in the developing and adult cerebral cortex and hippocampus  

Microsoft Academic Search

The temporal and spatial expression in brain of the mRNAs for the pleiotropic cytokine hepatocyte growth factor\\/scatter factor (HGF\\/SF) and its receptor c-met were compared to those of a known HGF\\/SF activator, tissue-type plasminogen activator (tPA). In addition to the previously described expression in the developing and adult olfactory system [D.P. Thewke, N.W. Seeds, Expression of hepatocyte growth factor\\/scatter factor,

Douglas P. Thewke; Nicholas W. Seeds



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



1,4-Bis(alkylamino)benzo[g]phthalazines able to form dinuclear complexes of Cu(II) which as free ligands behave as SOD inhibitors and show efficient in vitro activity against Trypanosoma cruzi.  


The synthesis of a new series of 1,4-bis(alkylamino)benzo[g]phthalazines 1-3 is reported, and their ability to form dinuclear complexes with Cu(II) assayed. The geometry of the complexes is dependent on the nature of the electron-donor sites at the sidechains. Compounds 1 and 2, that contain sp3 or sp2 nitrogens at the end of the alkylamino groups, originate monopodal dinuclear complexes which seem to include endogenous OH bridges, and the sidechains seem to actively participate in complexation. However, the substitution of nitrogen by oxygen in 3 leads to a tripodal dinuclear complex in which the sidechains are not involved. The in vitro antiparasitic activity on Trypanosoma cruzi epimastigotes and amastigotes and the SOD activity inhibition have been evaluated for compounds 1-3, and, as expected, 1 and 2 show in all cases relevant results, whereas 3 is always the less active among the three substrates tested. PMID:17222558

Rodríguez-Ciria, Marinela; Sanz, Ana M; Yunta, María J R; Gómez-Contreras, Fernando; Navarro, Pilar; Sánchez-Moreno, Manuel; Boutaleb-Charki, Samira; Osuna, Antonio; Castińeiras, Alfonso; Pardo, Mercedes; Cano, Carmen; Campayo, Lucrecia



Transcriptional Activation of the Aspergillus nidulans gpdA Promoter by Osmotic Signals†  

PubMed Central

A differentially expressed gpdA cDNA clone was isolated from NaCl-adapted Aspergillus nidulans (FGSC359) and identified as glyceraldehyde-3-phosphate dehydrogenase (gpdA) on the basis of its nucleotide sequence. The level of gpdA RNA substantially increased in cultures gradually adapted to NaCl but was greatly reduced in cultures exposed briefly to a high concentration of NaCl. A pyrG auxotroph of A. nidulans (A773) was cotransformed with a gpdA-uidA construct and a plasmid containing the Neurospora crassa pyr4 gene as a selectable marker. One pyrG+ ?-glucuronidase-positive (GUS+) transformant was selected, and stable integration of the gpdA-uidA construct into the genome was confirmed by Southern blot analysis. Gradual adaptation to increasing concentrations of NaCl resulted in an increase in GUS activity to 2.7-fold. GUS activity was reduced after a 2-h exposure of an unadapted culture to 2 M NaCl but gradually increased to a maximum of twofold after 24 h. GUS activity also increased by 8.4-fold in Na2SO4-adapted cultures, 4.9-fold in polyethylene glycol-adapted cultures, and 7.5-fold in KCl-adapted cultures. These results are consistent with the hypothesis that the A. nidulans gpdA promoter is transcriptionally activated by osmotic signals.

Redkar, Rajendra J.; Herzog, Roland W.; Singh, Narendra K.



Transcriptional activation of the Aspergillus nidulans gpdA promoter by osmotic signals.  


A differentially expressed gpdA cDNA clone was isolated from NaCl-adapted Aspergillus nidulans (FGSC359) and identified as glyceraldehyde-3-phosphate dehydrogenase (gpdA) on the basis of its nucleotide sequence. The level of gpdA RNA substantially increased in cultures gradually adapted to NaCl but was greatly reduced in cultures exposed briefly to a high concentration of NaCl. A pyrG auxotroph of A. nidulans (A773) was cotransformed with a gpdA-uidA construct and a plasmid containing the Neurospora crassa pyr4 gene as a selectable marker. One pyrG+ beta-glucuronidase-positive (GUS+) transformant was selected, and stable integration of the gpdA-uidA construct into the genome was confirmed by Southern blot analysis. Gradual adaptation to increasing concentrations of NaCl resulted in an increase in GUS activity to 2.7-fold. GUS activity was reduced after a 2-h exposure of an unadapted culture to 2 M NaCl but gradually increased to a maximum of twofold after 24 h. GUS activity also increased by 8.4-fold in Na2SO4-adapted cultures, 4.9-fold in polyethylene glycol-adapted cultures, and 7.5-fold in KCl-adapted cultures. These results are consistent with the hypothesis that the A. nidulans gpdA promoter is transcriptionally activated by osmotic signals. PMID:9603839

Redkar, R J; Herzog, R W; Singh, N K



Rapid Evolution of  Glucuronidase Specificity by Saturation Mutagenesis of an Active Site Loop  

Microsoft Academic Search

Protein engineers have widely adopted directed evo- lution as a design algorithm, but practitioners have not come to a consensus about the best method to evolve protein molecular recognition. We previously used DNA shuffling to direct the evolution of Escherichia coli -glucuronidase (GUS) variants with increased -galac- tosidase activity. Epistatic (synergistic) mutations in amino acids 557, 566, and 568, which

Melissa L. Geddie; Ichiro Matsumura



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.  


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. PMID:22003011

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



Activation and Repression of Transcription by Auxin-Response Factors  

Microsoft Academic Search

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

Tim Ulmasov; Gretchen Hagen; Tom J. Guilfoyle



View of hospital district, showing cannon in foreground, showing building ...  

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

View of hospital district, showing cannon in foreground, showing building H1 at left, showing building H72 in background; camera facing north. - Mare Island Naval Shipyard, East of Nave Drive, Vallejo, Solano County, CA


The promoter of the leghaemoglobin gene VfLb29: functional analysis and identification of modules necessary for its activation in the infected cells of root nodules and in the arbuscule-containing cells of mycorrhizal roots.  


In this study the further characterization of the Vicia faba leghaemoglobin promoter pVfLb29 is presented that was previously shown to be specifically active in the infected cells of root nodules and in arbuscule-containing cells of mycorrhizal roots. Using promoter studies in transgenic hairy roots of the Pisum sativum mutant RisNod24, disabled in the formation of functional arbuscules, VfLb29 promoter activity is assigned to later stages of arbuscule development. In order to narrow down the regions containing cis-acting elements of pVfLb29, the activity of five VfLb29 promoter deletions (-797/-31 to -175/-31 in relation to the start codon) fused to the gusAint coding region were tested in transgenic V. hirsuta hairy roots. The results specify a promoter region ranging from position -410 to -326 (85 bp) as necessary for gus expression in arbuscule-containing cells, whereas this segment is not involved in the nodule-specific activity. Sequence analysis of the pVfLb29 fragment -410/-326 (85 bp) revealed sequence motifs previously shown to be cis-acting elements of diverse promoters. To investigate the autonomous function of pVfLb29 regions for activation in arbuscule-containing cells, different regions of pVfLb29 from positions -410 to -198 were used to prepare chimeric promoter constructs for trans-activation studies. These fragments alone did not activate the mycorrhiza inactive promoter of the Vicia faba leghaemoglobin gene VfLb3, showing that the activation of pVfLb29 in arbuscule-containing cells is governed by a complex regulatory system that requires at least two modules located between position -410 and -31 of the VfLb29 gene. PMID:15668224

Fehlberg, Vera; Vieweg, Martin F; Dohmann, Esther M N; Hohnjec, Natalija; Pühler, Alfred; Perlick, Andreas M; Küster, Helge



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.  


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



A WRKY Transcription Factor Recruits the SYG1-Like Protein SHB1 to Activate Gene Expression and Seed Cavity Enlargement  

PubMed Central

Seed development in Arabidopsis and in many dicots involves an early proliferation of the endosperm to form a large embryo sac or seed cavity close to the size of the mature seed, followed by a second phase during which the embryo grows and replaces the endosperm. SHORT HYPOCOTYL UNDER BLUE1 (SHB1) is a member of the SYG1 protein family in fungi, Caenorhabditis elegans, flies, and mammals. SHB1 gain-of-function enhances endosperm proliferation, increases seed size, and up-regulates the expression of the WRKY transcription factor gene MINISEED3 (MINI3) and the LRR receptor kinase gene HAIKU2 (IKU2). Mutations in either IKU2 or MINI3 retard endosperm proliferation and reduce seed size. However, the molecular mechanisms underlying the establishment of the seed cavity and hence the seed size remain largely unknown. Here, we show that the expression of MINI3 and IKU2 is repressed before fertilization and after 4 days after pollination (DAP), but is activated by SHB1 from 2 to 4 DAP prior to the formation of the seed cavity. SHB1 associates with their promoters but without a recognizable DNA binding motif, and this association is abolished in mini3 mutant. MINI3 binds to W-boxes in, and recruits SHB1 to, its own and IKU2 promoters. Interestingly, SHB1, but not MINI3, activates transcription of pMINI3::GUS or pIKU2::GUS. We reveal a critical developmental switch through the activation of MINI3 expression by SHB1. The recruitment of SHB1 by MINI3 to its own and IKU2 promoters represents a novel two-step amplification to counter the low expression level of IKU2, which is a trigger for endosperm proliferation and seed cavity enlargement.

Kang, Xiaojun; Li, Wei; Zhou, Yun; Ni, Min



Expression of recombinant staphylokinase, a fibrin-specific plasminogen activator of bacterial origin, in potato (Solanum tuberosum L.) plants.  


One of the most dynamically developing sectors of green biotechnology is molecular farming using transgenic plants as natural bioreactors for the large scale production of recombinant proteins with biopharmaceutical and therapeutic values. Such properties are characteristic of certain proteins of bacterial origin, including staphylokinase. For many years, work has been carried out on the use of this protein in thrombolytic therapy. In this study, transgenic Solanum tuberosum plants expressing a CaMV::sak-mgpf-gusA gene fusion, were obtained. AGL1 A. tumefaciens strain was used in the process of transformation. The presence of the staphylokinase gene was confirmed by PCR in 22.5% of the investigated plants. The expression of the fusion transgene was detected using the ?-glucuronidase activity assay in 32 putative transgenic plants. Furthermore, on the basis of the GUS histochemical reaction, the transgene expression pattern had a strong, constitutive character in seven of the transformants. The polyacrylamide gel electrophoresis of a protein extract from the SAK/PCR-positive plants, revealed the presence of a119 kDa protein that corresponds to that of the fusion protein SAK-mGFP-GUSA. Western blot analysis, using an antibody against staphylokinase, showed the presence of the staphylokinase domain in the 119 kDa protein in six analyzed transformants. However, the enzymatic test revealed amidolytic activity characteristic of staphylokinase in the protein extract of only one plant. This is the first report on a Solanum tuberosum plant producing a recombinant staphylokinase protein, a plasminogen activator of bacterial origin. PMID:22347774

Gerszberg, Aneta; Wiktorek-Smagur, Aneta; Hnatuszko-Konka, Katarzyna; ?uchniak, Piotr; Kononowicz, Andrzej K



Role of cis-Acting Sites NorL, a TATA Box, and AflR1 in nor-1 Transcriptional Activation in Aspergillus parasiticus  

PubMed Central

The transcription factor AflR is required for up-regulation of specific pathway genes involved in aflatoxin biosynthesis in the filamentous fungus Aspergillus. nor-1 encodes an early aflatoxin pathway enzyme; its promoter contains a consensus AflR binding site (AflR1). Proteins in Aspergillus parasiticus cell extracts and AflR expressed in Escherichia coli do not bind to A. parasiticus AflR1 in vitro, so it was not clear if this site was required for nor-1 expression or if other transcription factors contributed to gene regulation. In this study we defined the role of AflR1 in nor-1 expression in A. parasiticus and identified additional cis-acting sites required for maximum nor-1 transcriptional activation. Deletion and substitution of AflR1 in the nor-1 promoter in A. parasiticus nor-1::GUS reporter strains showed that this site is required for nor-1 transcriptional activation in vivo. Substitution of a putative TATA box in the nor-1 promoter resulted in nondetectable ?-glucuronidase (GUS) activity, demonstrating that this TATA box is functional in vivo. We also identified a novel cis-acting site, designated NorL, between residues ?210 and ?238 that was required for maximum nor-1 transcriptional activation in A. parasiticus grown in liquid medium and on solid medium. Using an electrophoretic mobility shift assay, we identified a specific NorL-dependent DNA-protein complex that relies on a functional AflR, either directly or indirectly, for maximum binding capacity. Because the NorL site appears only once in the aflatoxin gene cluster, its association with the nor-1 promoter may have important implications for the overall regulatory scheme for the aflatoxin pathway.

Miller, Michael J.; Roze, Ludmila V.; Trail, Frances; Linz, John E.



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




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

28. MAP SHOWING LOCATION OF ARVFS FACILITY AS BUILT. SHOWS LINCOLN BOULEVARD, BIG LOST RIVER, AND NAVAL REACTORS FACILITY. F.C. TORKELSON DRAWING NUMBER 842-ARVFS-101-2. DATED OCTOBER 12, 1965. INEL INDEX CODE NUMBER: 075 0101 851 151969. - Idaho National Engineering Laboratory, Advanced Reentry Vehicle Fusing System, Scoville, Butte County, ID


Anatomically specific patterns of glial activation in the periaqueductal gray of the sub-population of rats showing pain and disability following chronic constriction injury of the sciatic nerve.  


Neuropathic pain conditions for which treatment is sought are characterized by complex behavioural disturbances, as well as "pain." Recent studies using chronic constriction injury of the sciatic nerve have shown that rats develop three distinct patterns of disability characterized by changes in social-interactions and sleep-wake cycle behaviours post-injury: (i) Persistent Disability, (ii) Transient Disability and (iii) No Disability. These patterns occur despite all rats showing identical levels of allodynia and hyperalgesia (i.e., pain). In rats, social-interactions and sleep-wake cycle behaviours are regulated in part, by neural networks, which converge on the periaqueductal grey (PAG). We sought therefore to identify neural adaptations in the PAG, 6 days following chronic constriction injury (CCI), the time at which rats in which disabilities persist are first distinguished from those without disabilities (i.e., No Disability and Transient Disability). GeneChips, RT-PCR and Western blotting revealed the select up-regulation in translation and transcription of glial fibrillary acidic protein (GFAP) and Vimentin in rats with Persistent Disability. Significant increases in GFAP immunoreactivity were localized histologically to the lateral and caudal ventrolateral columns of the PAG. This anatomically specific pattern of increased GFAP suggests activation of astrocytes by select neural pathways, which likely include afferents of both spinal and nucleus of the solitary tract (NTS) origin. The PAG columns in which astrocytes are activated play significant roles in modulating both social-interactions and the sleep-wake cycle. It is possible therefore that the persistent disabilities seen in a subgroup of CCI rats are in part a functional consequence of this specific pattern of astrocyte activation. PMID:20109535

Mor, D; Bembrick, A L; Austin, P J; Wyllie, P M; Creber, N J; Denyer, G S; Keay, K A



Synergistic Activation of the Pathogenicity-Related Proline Iminopeptidase Gene in Xanthomonas campestris pv. campestris by HrpX and a LuxR Homolog  

PubMed Central

Xanthomonas campestris pv. campestris strain 8004 contains an orphan quorum-sensing (QS) locus, xccR-pipXcc, in which the proline iminopeptidase (pipXcc) gene (where “Xcc” indicates that the pip gene is from X. campestris pv. campestris) is positively regulated by the LuxR homologue XccR by binding to the luxXc box of the pipXcc promoter. The disruption of pipXcc significantly attenuated the virulence of X. campestris pv. campestris. An imperfect plant-inducible promoter (PIP) box is located in the upstream region of the pipXcc promoter, which is the putative binding site of the transcriptional activator HrpX. To explore whether the expression of the pipXcc gene is regulated by HrpX, the expression level of a pipXcc promoter-gusA fusion gene was assayed in an hrpX disruption mutant. The results showed that the lack of HrpX dramatically decreased the ?-glucuronidase (GUS) activity. Further analyses using an electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP)-PCR indicated that the imperfect PIP box in X. campestris pv. campestris is specifically bound to HrpX. These data demonstrated that the pipXcc gene belongs to the hrp regulon and that the imperfect PIP box of the pipXcc promoter could be a cis element for the HrpX protein. We further showed in a pulldown assay that XccR can bind HrpX, suggesting that these two regulatory proteins coactivate the virulence factor by binding to the different cis elements of the pipXcc gene and adapt to the host environment during X. campestris pv. campestris infection.

Zhang, Jingxi; Kan, Jinhong; Zhang, Jieqiong; Guo, Ping; Chen, Xiaoying



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.



National Orange Show Photovoltaic Demonstration  

SciTech Connect

National Orange Show Photovoltaic Demonstration created a 400KW Photovoltaic self-generation plant at the National Orange Show Events Center (NOS). The NOS owns a 120-acre state fairground where it operates an events center and produces an annual citrus fair known as the Orange Show. The NOS governing board wanted to employ cost-saving programs for annual energy expenses. It is hoped the Photovoltaic program will result in overall savings for the NOS, help reduce the State's energy demands as relating to electrical power consumption, improve quality of life within the affected grid area as well as increase the energy efficiency of buildings at our venue. In addition, the potential to reduce operational expenses would have a tremendous effect on the ability of the NOS to service its community.

Dan Jimenez (NOS)Sheri Raborn, CPA (National Orange Show); Tom Baker (California Construction Authority)



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



Reading the traveling exhibition show: \\  

Microsoft Academic Search

This dissertation utilizes the motif of the traveling exhibition show in order to analyze how the Massachusetts Magazine (1789–96) participated in the cultural discussion regarding the construction of the American woman in the new nation. Although others have focused on the role of women in America (i.e., “Republican Motherhood”), I assert that whatever situation a woman found herself in—single, married,

Beverly Jean Reed



Do Trade Shows Pay Off?  

Microsoft Academic Search

rade show expenditures are the second largest item in the business marketing communications budget after advertising, and they account for nearly one-fifth of the total budget for U.S. firms and approximately one-fourth of the budget for European firms (Jacobson 1990; Schafer 1987). The level of these expenditures, including direct costs and allocation of exhibitor staff time, though exclud- ing planning

Srinath Gopalakrishna; Gary L. Lilien; Jerome D. Williams; Ian K. Sequeira


Rocks and Minerals Slide Show  

NSDL National Science Digital Library

This interactive slide show of common rocks and minerals allows students to choose from two sets of minerals and click on a thumbnail to see a larger photograph with a full description of the mineral including color, streak, hardness, cleavage/fracture, and chemical composition. Also included are its use and where it is found. The rocks are divided into igneous, sedimentary, and metamorphic and can be accessed in the same manner. They are described on the basis of crystal size and mineral composition as well as use.


Antibody-induced oligomerization and activation of an engineered reporter enzyme  

PubMed Central

Summary Our objective is to produce a protein biosensor (or molecular switch) that is specifically activated in solution by a monoclonal antibody. Many effector-dependent enzymes have evolved in nature, but the introduction of a novel regulatory mechanism into a normally unregulated enzyme poses a difficult design problem. We used site-saturation mutagenesis and screening to generate antibody-activated variants of the reporter enzyme beta-glucuronidase (GUS). The specific activity of the purified epitope-tagged GUS variant was increased by up to ~500-fold by the addition an equimolar concentration of a monoclonal antibody. This molecular switch is modular in design, so it can easily be re-engineered for the detection of other peptide-specific antibodies. Such antibody-activated reporters could someday enable point-of-care serological assays for the rapid detection of infectious diseases.

Geddie, Melissa L.; Matsumura, Ichiro



Computationally-predicted CB1 cannabinoid receptor mutants show distinct patterns of salt-bridges that correlate with their level of constitutive activity reflected in G protein coupling levels, thermal stability, and ligand binding.  


The cannabinoid receptor 1 (CB1), a member of the class A G-protein-coupled receptor (GPCR) family, possesses an observable level of constitutive activity. Its activation mechanism, however, has yet to be elucidated. Previously we discovered dramatic changes in CB1 activity due to single mutations; T3.46A, which made the receptor inactive, and T3.46I and L3.43A, which made it essentially fully constitutively active. Our subsequent prediction of the structures of these mutant receptors indicated that these changes in activity are explained in terms of the pattern of salt-bridges in the receptor region involving transmembrane domains 2, 3, 5, and 6. Here we identified key salt-bridges, R2.37 + D6.30 and D2.63 + K3.28, critical for CB1 inactive and active states, respectively, and generated new mutant receptors that we predicted would change CB1 activity by either precluding or promoting these interactions. We find that breaking the R2.37 + D6.30 salt-bridge resulted in substantial increase in G-protein coupling activity and reduced thermal stability relative to the wild-type reflecting the changes in constitutive activity from inactive to active. In contrast, breaking the D2.63 + K3.28 salt-bridge produced the opposite profile suggesting this interaction is critical for the receptor activation. Thus, we demonstrate an excellent correlation with the predicted pattern of key salt-bridges and experimental levels of activity and conformational flexibility. These results are also consistent with the extended ternary complex model with respect to shifts in agonist and inverse agonist affinity and provide a powerful framework for understanding the molecular basis for the multiple stages of CB1 activation and that of other GPCRs in general. PMID:23408552

Ahn, Kwang H; Scott, Caitlin E; Abrol, Ravinder; Goddard, William A; Kendall, Debra A



"Show me" bioethics and politics.  


Missouri, the "Show Me State," has become the epicenter of several important national public policy debates, including abortion rights, the right to choose and refuse medical treatment, and, most recently, early stem cell research. In this environment, the Center for Practical Bioethics (formerly, Midwest Bioethics Center) emerged and grew. The Center's role in these "cultural wars" is not to advocate for a particular position but to provide well researched and objective information, perspective, and advocacy for the ethical justification of policy positions; and to serve as a neutral convener and provider of a public forum for discussion. In this article, the Center's work on early stem cell research is a case study through which to argue that not only the Center, but also the field of bioethics has a critical role in the politics of public health policy. PMID:17926217

Christopher, Myra J



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)



Instrument Development for SHOW project  

NASA Astrophysics Data System (ADS)

Water is a critically important constituent throughout the stratosphere and mesosphere The SHOW project will develop a new instrument to measure water vapour from 15km to 85km height on a global scale using the unique capabilities provided by Spatial Heterodyne Spectroscopy SHS This work builds on Canadian expertise in fabricating solid Michelson interferometers to fill a significant niche in our current capability The SHS setup the FTS with the mirrors replaced by diffraction gratings at Littrow configuration wavelength depended Fizeau fringes are recorded by a 320 256 InGaAs near infrared camera without any scanning elements the high resolution spectral information along one detector dimension can be obtain from Fourier analysis and the other dimension will provide the spatial information At a limb view point a field-widened SHS with half-angle of 6 degrees for water observations at 1364nm is desired the resolution is 0 02nm within full bandwidth of 2nm and the resolving power is about 68 000 The laboratory work for the instrument development and the designing building and testing of the pre-prototype are presented

Lin, Y. L.; Shepherd, M. G.; Shepherd, G. G.; Solheim, B. H.; Brown, S.


Roles of salicylic acid-responsive cis-acting elements and W-boxes in salicylic acid induction of VCH3 promoter in transgenic tobaccos.  


A salicylic acid (SA)-inducible VCH3 promoter was recently identified from grapevine (Vitis amurensis) that contains two inverse SA-responsive cis-acting elements and four W-boxes. To further demonstrate the roles of these elements, four fragments with lengths from -1187, -892, -589, -276 to +7 bp were fused with the b-glucuronidase (GUS) reporter gene and transferred to Nicotiana tobacum, together with another four VCH3 promoter fragments with mutation in the two inverse SA-responsive elements. The functions of each promoter fragment were examined by analysis of GUS activity in the transgenic tobacco root treated with SA. Enhanced GUS activity was shown in the roots of transgenic tobaccos with the VCH3 (-1187)-GUS construct containing two SA-responsive cis-acting elements and four W-boxes. However, GUS activity directed by the VCH3 (-892)-GUS construct, containing one SA cis-acting element and four W-boxes, was reduced by up to 35% compared with that in tobaccos transformed with the VCH3 (-1187)-GUS construct, indicating that the SA cis-acting element plays an important role in SA induction of the VCH3 promoter. Neither the m2VCH3 (-1187)-GUS nor the mVCH3 (-892)-GUS construct, with mutation on the SA-responsive elements, abolished the expression of GUS activity, demonstrating that the W-boxes in the VCH3 promoter are also involved in SA induction. Histochemical analysis of GUS activity directed by each of the eight VCH3 promoter fragments showed that GUS was expressed specifically in vascular tissue. It was concluded that both the SA-responsive cis-acting elements and the W-boxes are important for the SA induction of the VCH3 promoter. This promoter might have a potential use in plant genetic engineering. PMID:16395526

Li, Hai-Yan; Wei, Wei; Li, Yu



A Talk Show from the Past.  

ERIC Educational Resources Information Center

|Describes a two-day activity in which elementary students examine voting rights, the right to assemble, and women's suffrage. Explains the game, "Assemble, Reassemble," and a student-produced talk show with five students playing the roles of leaders of the women's suffrage movement. Profiles Elizabeth Cady Stanton, Lucretia Mott, Susan B.…

Gallagher, Arlene F.



A Talk Show from the Past.  

ERIC Educational Resources Information Center

Describes a two-day activity in which elementary students examine voting rights, the right to assemble, and women's suffrage. Explains the game, "Assemble, Reassemble," and a student-produced talk show with five students playing the roles of leaders of the women's suffrage movement. Profiles Elizabeth Cady Stanton, Lucretia Mott, Susan B. Anthony,…

Gallagher, Arlene F.



Phytochrome-Mediated Photoperiod Perception, Shoot Growth, Glutamine, Calcium, and Protein Phosphorylation Influence the Activity of the Poplar Bark Storage Protein Gene Promoter (bspA)1  

PubMed Central

In poplars (Populus), bspA encodes a 32-kD bark storage protein that accumulates in the inner bark of plants exposed to either short-day (SD) photoperiods or elevated levels of nitrogen. In this study, poplars transformed with a chimeric gene consisting of the bspA promoter fused to ?-glucuronidase (uidA) were used to investigate the transcriptional regulation of the bspA promoter. Photoperiodic activation of the bspA promoter was shown to involve perception by phytochrome and likely involves both a low fluence response and a parallel very low fluence response pathway. Activity of the bspA promoter was also influenced by shoot growth. High levels of bspA expression usually occur in the bark of plants during SD but not long day or SD with a night break. When growth was inhibited under growth permissive photoperiods (SD with night break) levels of bark ?-glucuronidase (GUS) activity increased. Stimulating shoot growth in plants treated with SD inhibited SD-induced increases in bark GUS activity. Because changes in photoperiod and growth also alter carbon and nitrogen partitioning, the role of carbon and nitrogen metabolites in modulating the activity of the bspA promoter were investigated by treating excised stems with amino acids or NH4NO3 with or without sucrose. Treatment with either glutamine or NH4NO3 resulted in increased stem GUS activity. The addition of sucrose with either glutamine or NH4NO3 resulted in synergistic induction of GUS, whereas sucrose alone had no effect. Glutamine plus sucrose induction of GUS activity was inhibited by EGTA, okadaic acid, or K-252A. Inhibition by EGTA was partially relieved by the addition of Ca2+. The Ca2+ ionophore, ionomycin, also induced GUS activity in excised shoots. These results indicate that transcriptional activation of bspA is complex. It is likely that SD activation of bspA involves perception by phytochrome coupled to changes in growth. These growth changes may then alter carbon and nitrogen partitioning that somehow signals bspA induction by a yet undefined mechanism that involves carbon and nitrogen metabolites, Ca2+, and protein phosphorylation/dephosphorylation.

Zhu, Baolong; Coleman, Gary D.



Substitution of active-site His-223 in Pseudomonas aeruginosa elastase and expression of the mutated lasB alleles in Escherichia coli show evidence for autoproteolytic processing of proelastase.  

PubMed Central

The neutral metalloprotease elastase is one of the major proteins secreted into the culture medium by many Pseudomonas aeruginosa strains. Encoded by the lasB gene, the 33-kDa elastase is initially synthesized as a 53-kDa preproenzyme which is processed to the mature form via a 51-kDa proelastase intermediate. To facilitate studies on proteolytic processing of elastase precursors and on secretion, we developed systems for overexpression of lasB in Escherichia coli under the control of the inducible T7 and tac promoters. Although the 51-kDa proelastase form was detectable in E. coli under inducible conditions, most of the elastase produced under these conditions was found in an enzymatically active 33-kDa form. The amino-terminal sequence of the first 15 amino acid residues of this 33-kDa elastase species was identical to that of the mature P. aeruginosa enzyme, suggesting that processing was autocatalytic. To test this possibility, the codon in lasB encoding His-223, a presumed active-site residue, was changed to encode Asp-223 (lasB1) and Tyr-223 (lasB2). The effects of these mutations on enzyme activity and processing were examined. No proteolytic or elastolytic activities were detected in extracts of E. coli cells containing the lasB mutant alleles. Overexpression of the mutated lasB genes in E. coli resulted in the accumulation of the corresponding 51-kDa proelastase species. These were processed in vitro to the respective 33-kDa forms by incubation with exogenous purified elastase, without an increase in proteolytic activity. Molecular modeling studies suggest that the mutations have little or no effect on the conformation of the mutant elastases. In addition, wild-type elastase and the mutant proelastases were localized to the periplasm of E. coli. The present results confirm that His-223 is essential for elastase activity and provide evidence for autoproteolytic processing of proelastase. Images FIG. 2 FIG. 4A FIG. 4B FIG. 4C FIG. 5 FIG. 6

McIver, K; Kessler, E; Ohman, D E



Generation of a Stable Antioxidant Response Element-Driven Reporter Gene Cell Line and Its Use to Show Redox-Dependent Activation of Nrf2 by Cancer Chemotherapeutic Agents  

Microsoft Academic Search

The NF-E2 p45-related factor 2 (Nrf2) regulates cytoprotective genes that contain an antioxidant response element (ARE) in their promoters. To investigate whether anticancer drugs can induce ARE-driven gene expression, we have developed a stable human mammary MCF7-derived reporter cell line called AREc32, which contains a luciferase gene construct controlled by eight copies of the cis-element. In these cells, luciferase activity

Xiu Jun Wang; John D. Hayes



Erythroid Cells Rendered Erythropoietin Independent by Infection with Friend Spleen Focus-Forming Virus Show Constitutive Activation of Phosphatidylinositol 3Kinase and Akt Kinase: Involvement of Insulin Receptor Substrate-Related Adapter Proteins  

Microsoft Academic Search

The erythroleukemia-inducing Friend spleen focus-forming virus (SFFV) encodes a unique envelope glyco- protein which allows erythroid cells to proliferate and differentiate in the absence of erythropoietin (Epo). In an effort to understand how SFFV causes Epo independence, we have been examining erythroid cells rendered factor independent by SFFV infection for constitutive activation of signal-transducing molecules. Previous studies from our laboratory




Human breast cancer MDA-MB-231 cells fail to express the neurofibromin protein, lack its type I mRNA isoform and show accumulation of P-MAPK and activated Ras.  


Neurofibromin is a tumor suppressor protein, which is similar in function to the GTPase activating protein (GAP), p120GAP, in that it accelerates inactivation of Ras. Mutations in the NF1 gene cause neurofibromatosis type 1, NF1, an autosomal dominant disease with a diverse spectrum of clinical manifestations, including neurofibromas. Ras activation (GTP binding) is induced by the GTP exchange factor Sos and its inactivation is regulated through the GAPs (p120GAP and neurofibromin). Strikingly, neurofibromin was nearly absent in MB-231 human breast cancer cells and present in the remaining four cell lines studied, with higher levels in BT-474 and MB-453 than in MCF-7 and BT-20 cells, as tested with polyclonal antibodies to both the N-terminal as well as the C-terminal peptides. Coordinated with the near absence of neurofibromin, these cells also presented with much greater levels of P-MAPK and activated Ras. Further, RT-PCR analysis demonstrated the absence of expression of NF1 mRNA type I isoform only in the MB-231 cell lines. This result documents for the first time an altered NF1 expression at the protein and mRNA levels in MDA-MB-231 breast cancer cells. PMID:11566491

Ogata, H; Sato, H; Takatsuka, J; De Luca, L M



The oestrogenic and anti-platelet activities of dihydrobenzofuroisocoumarins and homoisoflavonoids from Liriope platyphylla roots.  


The ethanolic extract of Liriope platyphylla (Liliaceae) roots showed potential oestrogenic and anti-platelet activities. Twenty-six compounds were isolated and classified as 10 skeletons, including two unusual new dihydrobenzofuroisocoumarins, (+)-platyphyllarin A (1) and B (2), one new butanoate, ethyltributanoate (3), and two new homoisoflavanones, (-)-liriopein A (4) and B (5), along with 21 known compounds, including six homoisoflavonoids, one chalcone, six amides, one lignan, one fatty acid derivative, one alkaloid, three benzenoids, and two steroids. The biosynthetic pathway of compounds 1 and 2 was proposed in the current investigation. The oestrogenic activity of the isolates was evaluated utilising the pER8:GUS reporter assay system in transgenic Arabidopsis plant as well as the SEAP reporter assay system in MCF-7 breast cancer cell-line; the anti-platelet activity was evaluated using the anti-platelet aggregation assay. Several components exhibited significant oestrogenic and anti-platelet activities; demonstrating for the first time the potential use of L. platyphylla as a nutritional supplement for cardiovascular and endocrine diseases. PMID:23578647

Tsai, Yu-Chi; Chiang, Shang-Yu; El-Shazly, Mohamed; Wu, Chin-Chung; Beerhues, Ludger; Lai, Wan-Chun; Wu, Shou-Fang; Yen, Ming-Hong; Wu, Yang-Chang; Chang, Fang-Rong



Antitumor Activity of NVP-BKM120-A Selective Pan Class I PI3 Kinase Inhibitor Showed Differential Forms of Cell Death Based on p53 Status of Glioma Cells  

PubMed Central

Purpose The aim of this study was to show preclinical efficacy and clinical development potential of NVP-BKM120, a selective pan class I phosphatidylinositol-3 kinase (PI3K) inhibitor in human glioblastoma (GBM) cells in vitro and in vivo. Experimental Design The effect of NVP-BKM120 on cellular growth was assessed by CellTiter-Blue assay. Flow cytometric analyses were carried out to measure the cell-cycle, apoptosis, and mitotic index. Mitotic catastrophe was detected by immunofluorescence. The efficacy of NVP-BKM120 was tested using intracranial U87 glioma model. Results We tested the biologic effects of a selective PI3K inhibitor NVP-BKM120 in a set of glioma cell lines. NVP-BKM120 treatment for 72 hours resulted in a dose-dependent growth inhibition and effectively blocked the PI3K/Akt signaling cascade. Although we found no obvious relationship between the cell line's sensitivity to NVP-BKM120 and the phosphatase and tensin homolog (PTEN) and epidermal growth factor receptor (EGFR) statuses, we did observe a differential sensitivity pattern with respect to p53 status, with glioma cells containing wild-type p53 more sensitive than cells with mutated or deleted p53. NVP-BKM120 showed differential forms of cell death on the basis of p53 status of the cells with p53 wild-type cells undergoing apoptotic cell death and p53 mutant/deleted cells having a mitotic catastrophe cell death. NVP-BKM120 mediates mitotic catastrophe mainly through Aurora B kinase. Knockdown of p53 in p53 wild-type U87 glioma cells displayed microtubule misalignment, multiple centrosomes, and mitotic catastrophe cell death. Parallel to the assessment of the compound in in vitro settings, in vivo efficacy studies using an intracranial U87 tumor model showed an increased median survival from 26 days (control cohort) to 38 and 48 days (treated cohorts). Conclusion Our present findings establish that NVP-BKM120 inhibits the PI3K signaling pathways, leading to different forms of cell death on the basis of p53 statuses. Further studies are warranted to determine if NVP-BKM120 has potential as a glioma treatment.

Koul, Dimpy; Fu, Jun; Shen, Ruijun; LaFortune, Tiffany A.; Wang, Shuzhen; Tiao, Ningyi; Kim, Yong-Wan; Liu, Juinn-Lin; Ramnarian, Deepti; Yuan, Ying; Garcia-Echevrria, Carlos; Maira, Sauveur-Michel; Yung, W.K. Alfred



Show that a Quadrilateral is a Parallelogram  

NSDL National Science Digital Library

This essentially goes backwards; if certain conditions are met, then a quadrilateral can be shown to be a paralleogram. Find the proofs for Theorems 8.7, 8.8, 8.9, and 8.10 in the McDougall Littell text on pages 522 - 524 (go to and login): Class Zone Geometry Textbook Now, find the box that shows five ways to prove that a quadrilateral is a parallelogram (page 525 in the text): Class Zone Geometry Textbook Here is an activity to see if ...

Neubert, Mrs.



The universal bundling activity of AtVLN4 in diffusely growing cells.  


We recently reported that AtVLN4, a member of villin/gelsolin/fragmin superfamily in Arabidopsis thaliana, participated in root hair growth through its actin bundling activity. To further understand the functions of AtVLN4, we investigated its in vivo expression pattern and roles in diffusely growing cells. Transcription analysis of AtVLN4 and detection of AtVLN 4 promoter-GUS activity consistently indicated that AtVLN4 had a universal expression pattern and was preferentially expressed in vegetative tissues. Observation of actin structures labeled by GFP-fABD2 revealed that there were less actin bundles in many diffusely growing cell types in atvln4-1 seedlings than in wild-type seedlings. Pharmacological studies by treatment with Latrunculin B showed that the actin filaments were much easier to be disrupted in diffusely growing cells of atvln4-1 seedlings. Collectively, these results demonstrate that AtVLN4 has a universal actin bundling activity in diffusely growing cells just like that in the tip growing cell, root hairs. PMID:22019634

Du, Fei; Zhang, Yi; Ren, Haiyun



Ancient bacteria show evidence of DNA repair  

PubMed Central

Recent claims of cultivable ancient bacteria within sealed environments highlight our limited understanding of the mechanisms behind long-term cell survival. It remains unclear how dormancy, a favored explanation for extended cellular persistence, can cope with spontaneous genomic decay over geological timescales. There has been no direct evidence in ancient microbes for the most likely mechanism, active DNA repair, or for the metabolic activity necessary to sustain it. In this paper, we couple PCR and enzymatic treatment of DNA with direct respiration measurements to investigate long-term survival of bacteria sealed in frozen conditions for up to one million years. Our results show evidence of bacterial survival in samples up to half a million years in age, making this the oldest independently authenticated DNA to date obtained from viable cells. Additionally, we find strong evidence that this long-term survival is closely tied to cellular metabolic activity and DNA repair that over time proves to be superior to dormancy as a mechanism in sustaining bacteria viability.

Johnson, Sarah Stewart; Hebsgaard, Martin B.; Christensen, Torben R.; Mastepanov, Mikhail; Nielsen, Rasmus; Munch, Kasper; Brand, Tina; Gilbert, M. Thomas P.; Zuber, Maria T.; Bunce, Michael; R?nn, Regin; Gilichinsky, David; Froese, Duane; Willerslev, Eske



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


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



In plant activation: an inducible, hyperexpression platform for recombinant protein production in plants.  


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. PMID:23839786

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



Tagged and untagged TRAIL show different activity against tumor cells.  


The tumor necrosis factor-related apoptosis-inducing ligand (TRAIL/Apo2L) is a novel cytotoxic ligand belonging to the TNF superfamily which is currently being developed as a cancer therapeutic drug. Here, we observed the different functions of recombinant TRAIL protein with a foreign protein label and non-labeled TRAIL. We used a prokaryotic expression system to prepare two different versions of the extracellular TRAIL 114-281aa protein: TRAIL-HS, a protein modified with 6xHis-Tag and S-Tag; and TRAIL-FT, which had no foreign protein. The proteins were purified using Ni-NTA chromatography (TRAIL-HS) and cation ion-exchange column chromatography (TRAIL-FT) and identified by SDS-PAGE and western blot analysis. We compared the abilities of the proteins to bind to death receptor 5 (DR5) by ELISA and to induce apoptosis in a normal liver cell line (Chang liver) and a human T-lymphocyte leukemia cell line (Jurkat) by MTT assay, GR staining and FACS. The results indicate that the biological functions of TRAIL-FT were superior to those of TRAIL-HS in binding and the induction of apoptosis, and may be useful to further the development and applications of TRAIL. PMID:23205127

Zhao, Kunpeng; Wang, Yan'ge; Wang, Xueyin; Wang, Yugang; Ma, Yuanfang



The activity of the coat protein promoter of chloris striate mosaic virus is enhanced by its own and C1-C2 gene products.  


DNA fragments from the bidirectional promoter region of the geminivirus chloris striate mosaic virus (CSMV) were cloned into the pUC18-based vector, pG1 producing transcriptional fusions with the beta-glucuronidase (GUS) gene and nopaline synthase terminator sequence. The relative activity of each promoter construct was analyzed by a GUS expression assay of extracts from Zea mays (maize) Black Mexican Sweet protoplasts coelectroporated with the GUS reporter constructs and constructs in which individual CSMV open reading frames (ORFs) were placed under control of a cauliflower mosaic virus 35 S promoter. Weak promoter activity was observed for the promoter of the C1 and C2 ORFs (C1-C2 gene) and for the promoter of the V1 ORF. The activity of these promoters was unaffected by coelectroporation with the CSMV ORF constructs. Moderate activity was observed for the promoter of the V2 ORF (coat protein gene) which was enhanced by coelectroporation of the C1-C2 ORF construct. Sequences within the C1-C2 gene responsible for transactivation of the V2 ORF promoter were mapped close to the A site of a conserved NTP-binding sequence pattern within the C2 ORF. To a lesser extent activity for the promoter of the V2 ORF was enhanced by the V2 ORF construct providing evidence for positive autoregulation of the CSMV coat protein gene. PMID:8438584

Zhan, X; Richardson, K A; Haley, A; Morris, B A



Promoter activity associated with the intergenic regions of banana bunchy top virus DNA-1 to -6 in transgenic tobacco and banana cells.  


Promoter regions associated with each of the six ssDNA components of banana bunchy top virus (BBTV) have been characterized. DNA segments incorporating the intergenic regions of BBTV DNA-1 to -6 were isolated and fused to the uidA (beta-glucuronidase) reporter gene to assess promoter activity. In tobacco cell suspensions, the BBTV DNA-2 and -6 promoters generated levels of GUS expression 2-fold greater and similar to the 800 bp CaMV 35S promoter, respectively. Deletion analysis of the BBTV DNA-6 promoter suggested all the necessary promoter elements required for strong expression were located within 239 nucleotides upstream of the translational start codon. In transgenic tobacco plants, the BBTV-derived promoters generally provided a weak, tissue-specific GUS expression pattern restricted to phloem-associated cells. However, in callus derived from tobacco leaf tissue, GUS expression directed by the BBTV DNA-6 promoter was strong and, in some lines, comparable to the CaMV 35S promoter. Detectable promoter activity associated with the BBTV promoters in banana embryogenic cells was only observed using a sensitive green fluorescent protein (GFP) reporter. Promoters derived from BBTV DNA-4 and -5 generated the highest levels of transient activity, which were greater than that of the maize ubi-1 promoter. In transgenic banana plants, the activity of the BBTV DNA-6 promoter was restricted to the phloem of leaves and roots, stomata and root meristems. PMID:9780033

Dugdale, B; Beetham, P R; Becker, D K; Harding, R M; Dale, J L



Permanent prostate implant using high activity seeds and inverse planning with fast simulated annealing algorithm: A 12-year Canadian experience  

SciTech Connect

Purpose: To report outcomes and toxicity of the first Canadian permanent prostate implant program. Methods and Materials: 396 consecutive patients (Gleason {<=}6, initial prostate specific antigen (PSA) {<=}10 and stage T1-T2a disease) were implanted between June 1994 and December 2001. The median follow-up is of 60 months (maximum, 136 months). All patients were planned with fast-simulated annealing inverse planning algorithm with high activity seeds ([gt] 0.76 U). Acute and late toxicity is reported for the first 213 patients using a modified RTOG toxicity scale. The Kaplan-Meier biochemical failure-free survival (bFFS) is reported according to the ASTRO and Houston definitions. Results: The bFFS at 60 months was of 88.5% (90.5%) according to the ASTRO (Houston) definition and, of 91.4% (94.6%) in the low risk group (initial PSA {<=}10 and Gleason {<=}6 and Stage {<=}T2a). Risk factors statistically associated with bFFS were: initial PSA >10, a Gleason score of 7-8, and stage T2b-T3. The mean D90 was of 151 {+-} 36.1 Gy. The mean V100 was of 85.4 {+-} 8.5% with a mean V150 of 60.1 {+-} 12.3%. Overall, the implants were well tolerated. In the first 6 months, 31.5% of the patients were free of genitourinary symptoms (GUs), 12.7% had Grade 3 GUs; 91.6% were free of gastrointestinal symptoms (GIs). After 6 months, 54.0% were GUs free, 1.4% had Grade 3 GUs; 95.8% were GIs free. Conclusion: The inverse planning with fast simulated annealing and high activity seeds gives a 5-year bFFS, which is comparable with the best published series with a low toxicity profile.

Martin, Andre-Guy [Departement de Radio-oncologie, Centre de Recherche en Cancerologie de l'Universite Laval, Quebec (Canada); Roy, Jean [Departement de Radio-oncologie, Centre de Recherche en Cancerologie de l'Universite Laval, Quebec (Canada); Beaulieu, Luc [Departement de Radio-oncologie, Centre de Recherche en Cancerologie de l'Universite Laval, Quebec (Canada); Centre de Recherche de l'Hotel-Dieu de Quebec, L'Hotel-Dieu de Quebec, Centre Hospitalier Universitaire de Quebec (CHUQ), Centre de Recherche en Cancerologie de l'Universite Laval, Quebec (Canada); Pouliot, Jean [Radiation Oncology, University of California San Francisco, San Francisco, CA (United States); Harel, Francois [Centre de Recherche de l'Hotel-Dieu de Quebec, L'Hotel-Dieu de Quebec, Centre Hospitalier Universitaire de Quebec (CHUQ), Centre de Recherche en Cancerologie de l'Universite Laval, Quebec (Canada); Vigneault, Eric [Departement de Radio-oncologie, Centre de Recherche en Cancerologie de l'Universite Laval, Quebec (Canada)]. E-mail:



The promoter of the Arabidopsis thaliana BAN gene is active in proanthocyanidin-accumulating cells of the Brassica napus seed coat  

Microsoft Academic Search

As part of an ongoing research program dedicated to the understanding of proanthocyanidin (PA) accumulation in Brassica napus seed coat, transgenic rapeseed plants carrying a 2.3-kb fragment of the Arabidopsis thaliana BAN promoter (ProAtBAN) fused to the uidA reporter gene (GUS) were generated. Analysis of these plants revealed that ProAtBAN was activated in B. napus seed coat, following a spatio-temporal

Nathalie Nesi; Marie-Odile Lucas; Bathilde Auger; Cécile Baron; Alain Lécureuil; Philippe Guerche; Jocelyne Kronenberger; Loďc Lepiniec; Isabelle Debeaujon; Michel Renard



Promoter activity associated with the intergenic regions of banana bunchy top virus DNA1 to -6 in transgenic tobacco and banana cells  

Microsoft Academic Search

Promoter regions associated with each of the six ssDNA components of banana bunchy top virus (BBTV) have been characterized. DNA segments incorporating the intergenic regions of BBTV DNA-1 to -6 were isolated and fused to the uidA (b- glucuronidase) reporter gene to assess promoter activity. In tobacco cell suspensions, the BBTV DNA- 2 and -6 promoters generated levels of GUS

Benjamin Dugdale; Peter R. Beetham; Douglas K. Becker; Robert M. Harding; James L. Dale



The 'cleavage' activities of foot-and-mouth disease virus 2A site-directed mutants and naturally occurring '2Alike' sequences  

Microsoft Academic Search

The 2A\\/2B cleavage of aphtho- and cardiovirus 2A polyproteins is mediated by their 2A proteins 'cleaving' at their own C termini. We have analysed this activity using artificial reporter polyprotein systems comprising green fluorescent protein (GFP) linked via foot-and-mouth disease virus (FMDV) 2A to b-glucuronidase (GUS) - forming a single, long, open reading frame. Analysis of the distribution of radiolabel

Michelle L. L. Donnelly; Lorraine E. Hughes; Garry Luke; Heidi Mendoza; David Gani; Martin D. Ryan



The great American medicine show revisited.  


Since the late 1800s, changes in the advertising and marketing of medicinal drugs have produced heated debates in the United States. With the emergence of the modern prescription drug between 1938 and 1951, concerns that once focused primarily on patients' use of over-the-counter drugs were broadened to include physicians and their "doctors' drugs" as well. The medical profession's growing control over their patients' drug choices inevitably heightened the scrutiny of their own performance as consumers. Although deeply divided over issues of the patient's role in medical decision making, consumer activists and physician reformers expressed similar concerns about the impact of aggressive pharmaceutical marketing and advertising on the doctor-patient relationship, and starting in the late 1950s they employed strikingly similar strategies to counter the new corporate "medicine show." Yet their efforts to promote a more rational use of prescription drugs have usually been too little and too late to offset the effectiveness of pharmaceutical advertising and mar-keting activities. PMID:16327082

Tomes, Nancy



The promoter of the Vicia faba L. leghemoglobin gene VfLb29 is specifically activated in the infected cells of root nodules and in the arbuscule-containing cells of mycorrhizal roots from different legume and nonlegume plants.  


The VfLb29 leghemoglobin gene promoter was polymerase chain reaction-amplified from a Vicia faba genomic library and was fused to the gusAint coding region. Expression of the chimeric gene was analyzed in transgenic hairy roots of the legumes V. faba, V. hirsuta, and Medicago truncatula as well as in transgenic Nicotiana tabacum plants. The VfLb29 promoter was found to be specifically active not only in the infected cells of the nitrogen-fixing zone of root nodules but also in arbuscule-containing cells of transgenic V. faba and M. truncatula roots colonized by the endomycorrhizal fungus Glomus intraradices. In addition to these two legumes, specific expression in arbuscule-containing cells was also observed in the nonlegume N. tabacum. All studies were done in comparison to the V. faba leghemoglobin gene promoter VfLb3 that as VfLb29 was expressed in the infected cells of root nodules but showed no activity in endomycorrhiza. An activation of the VfLb29 promoter due to hypoxia in metabolically active tissues was excluded. The conserved activation in arbuscule-containing cells of legumes and the nonlegume N. tabacum suggests a conserved trigger for this promoter in legume and nonlegume endomycorrhiza symbioses. PMID:14714869

Vieweg, Martin F; Frühling, Martin; Quandt, Hans-Joachim; Heim, Ute; Bäumlein, Helmut; Pühler, Alfred; Küster, Helge; Andreas, M Perlick



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



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.



Silicon nanowire detectors showing phototransistive gain  

Microsoft Academic Search

Nanowire photodetectors are shown to function as phototransistors with high sensitivity. Due to small lateral dimensions, a nanowire detector can have low dark current while showing large phototransistive gain. Planar and vertical silicon nanowire photodetectors fabricated in a top-down approach using an etching process show a phototransistive gain above 35 000 at low light intensities. Simulations show that incident light

Arthur Zhang; Cesare Soci; Yisi Liu; Deli Wang; Yu-Hwa Lo



Watching The Daily Show in Kenya  

Microsoft Academic Search

Global distribution of a popular American television programme – Jon Stewart's Daily Show – offers a rare opportunity to examine transnational contingencies of meaning in political satire. Drawing on focus group discussions in Kenya, this analysis shows how some East Africans appropriated and reinterpreted – indeed unexpectedly subverted – The Daily Show's political content, deriving from it insights that Stewart

Angelique Haugerud; Dillon Mahoney; Meghan Ference



Talk shows’ representations of interpersonal conflicts  

Microsoft Academic Search

In the past ten years, daytime talk shows became very popular among television programmers and viewers alike. Given the large audiences to whom talk shows communicate, it is important to analyze the messages contained in the programs. Remarkably little academic attention has been paid to this phenomenon, however. The present study focuses on the presentation of interpersonal conflicts, particularly regarding

Susan L. Brinson; J. Emmett Winn



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.


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



Acculturation, Cultivation, and Daytime TV Talk Shows.  

ERIC Educational Resources Information Center

|Explores the cultivation phenomenon among international college students in the United States by examining the connection between levels of acculturation, daytime TV talk show viewing, and beliefs about social reality. Finds that students who scored low on acculturation and watched a great deal of daytime talk shows had a more negative perception…

Woo, Hyung-Jin; Dominick, Joseph R.



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…

Sergel, Sherman Louis, Ed.


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).  

PubMed Central

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, chlorophyll contents, and cell 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 green sectors to compensate for a lack of 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.

Meehan, L.; Harkins, K.; Chory, J.; Rodermel, S.



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



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



Best in Show: Teaching Old Dogs to Use New Rubrics  

ERIC Educational Resources Information Center

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 "The Dog Show Analogy." After participating in the activity, we have observed that the participating preservice…

Hitt, Austin M.; Helms, Emory C.



Silicon nanowire detectors showing phototransistive gain  

NASA Astrophysics Data System (ADS)

Nanowire photodetectors are shown to function as phototransistors with high sensitivity. Due to small lateral dimensions, a nanowire detector can have low dark current while showing large phototransistive gain. Planar and vertical silicon nanowire photodetectors fabricated in a top-down approach using an etching process show a phototransistive gain above 35 000 at low light intensities. Simulations show that incident light can be waveguided into vertical nanowires resulting in up to 40 times greater external quantum efficiency above their physical fill factor. Vertical silicon nanowire phototransistors formed by etching are attractive for low light level detection and for integration with silicon electronics.

Zhang, Arthur; You, Sifang; Soci, Cesare; Liu, Yisi; Wang, Deli; Lo, Yu-Hwa



Isolation and identification of a super strong plant promoter from cotton leaf curl Multan virus  

Microsoft Academic Search

The activity of the C1 and the V1 gene promoter of cotton leaf curl Multan virus (CLCuMV) was investigated in transgenic plants with the gus gene as a reporter gene. Quantitative GUS activity analysis of the transgenic plant leaves showed the average activity of the CLCuMV C1 gene promoter was 3- to 5-fold higher than that of the CaMV 35S

Yingqiu Xie; Yule Liu; Meng Meng; Lei Chen; Zhen Zhu



47 CFR 90.505 - Showing required.  

Code of Federal Regulations, 2011 CFR

...PRIVATE LAND MOBILE RADIO SERVICES Developmental Operation § 90.505 Showing required...this section, each application for developmental operation shall be accompanied by a...apply when an application is made for developmental operation solely for the reason...



47 CFR 90.505 - Showing required.  

Code of Federal Regulations, 2012 CFR

...PRIVATE LAND MOBILE RADIO SERVICES Developmental Operation § 90.505 Showing required...this section, each application for developmental operation shall be accompanied by a...apply when an application is made for developmental operation solely for the reason...



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. ...



Stutterers Show Different Brain Development, Study Says  


... this page, please enable JavaScript. Stutterers Show Different Brain Development, Study Says Less gray matter seen in ... have less gray matter in regions of the brain responsible for speech than those who don't ...


FDA Broadcast Shows Food Industry Personnel and ...  

Center for Food Safety and Applied Nutrition (CFSAN)

... FDA Broadcast Shows Food Industry Personnel and Consumers How Proper Health and Hygiene Helps to Prevent Foodborne Illness Outbreaks. ... More results from


Do dogs (Canis familiaris) show contagious yawning?  


We report an experimental investigation into whether domesticated dogs display contagious yawning. Fifteen dogs were shown video clips of (1) humans and (2) dogs displaying yawns and open-mouth expressions (not yawns) to investigate whether dogs showed contagious yawning to either of these social stimuli. Only one dog performed significantly more yawns during or shortly after viewing yawning videos than to the open-mouth videos, and most of these yawns occurred to the human videos. No dogs showed significantly more yawning to the open-mouth videos (human or dog). The percentage of dogs showing contagious yawning was less than chimpanzees and humans showing this behavior, and considerably less than a recently published report investigating this behavior in dogs (Joly-Mascheroni et al. in Biol Lett 4:446-448, 2008). PMID:19452178

Harr, Aimee L; Gilbert, Valerie R; Phillips, Kimberley A



Terpanes from oil shows of Lake Baikal  

Microsoft Academic Search

The composition and distribution of terpanes in oil shows that emerge from the bottom of Lake Baikal were studied. The oil\\u000a samples from three sites of Baikal showed different extents of their bacterial degradation, from practically unaltered to\\u000a heavily degraded oils. In the unaltered samples of oils and moderately degraded differences, a unique variety of biomarkers\\u000a (sesquiterpanes, secohopanes, new regular

V. A. Kashirtsev; A. E. Kontorovich; V. I. Moskvin; V. P. Danilova; V. N. Melenevskii



Cartograms showing China's population and wealth distribution  

Microsoft Academic Search

Population and wealth distributions are two important indicators of socioeconomic activities in a country. Understanding the spatial nature of population distribution is crucial for effective resource allocation and reasonable economic and social policies, factors that influence every aspect of everyday life. Contemporary China is characterized by very rapid economic development and profound social changes, many of them involving population distribution

Linna Li; Keith C. Clarke



TV Talk: Instant Classroom Celebrities With TV Game Shows  

ERIC Educational Resources Information Center

|Suggests some learning activities that can be used to capitalize on the popularity of TV game shows so that students can build their learning skills. These game show formats can also strengthen and reinforce many areas of the elementary curriculum. (Author/RK)|

Sheppard, Marti



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.



Expression of Foreign Genes in Transgenic Yellow-Poplar Plants 1  

PubMed Central

Cells of yellow-poplar (Liriodendron tulipifera L.) were transformed by direct gene transfer and regenerated into plants by somatic embryogenesis. Plasmid DNA bearing marker genes encoding ?-glucuronidase (GUS) and neomycin phosphotransferase (NPT II) were introduced by microprojectile bombardment into single cells and small cell clusters isolated from embryogenic suspension cultures. The number of full-length copies of the GUS gene in independently transformed callus lines ranged from approximately 3 to 30. An enzyme-linked immunosorbent assay for NPT II and a fluorometric assay for GUS showed that the expression of both enzymes varied by less than fourfold among callus lines. A histochemical assay for GUS activity revealed a heterogeneous pattern of staining with the substrate 5-bromo-4-chloro-3-indoyl-?-d-glucuronic acid in some transformed cell cultures. However, cell clusters reacting positively (blue) or negatively (white) with 5-bromo-4-chloro-3-indoyl-?-d-glucuronic acid demonstrated both GUS activity and NPT II expression in quantitative assays. Somatic embryos induced from transformed cell cultures were found to be uniformly GUS positive by histochemical analysis. All transgenic plants sampled expressed the two marker genes in both root and shoot tissues. GUS activity was found to be higher in leaves than roots by fluorometric and histochemical assays. Conversely, roots expressed higher levels of NPT II than leaves. ImagesFigure 1Figure 2

Wilde, H. Dayton; Meagher, Richard B.; Merkle, Scott A.



Experiments showing dynamics of materials interfaces  

SciTech Connect

The discipline of materials science and engineering often involves understanding and controlling properties of interfaces. The authors address the challenge of educating students about properties of interfaces, particularly dynamic properties and effects of unstable interfaces. A series of simple, inexpensive, hands-on activities about fluid interfaces provides students with a testbed to develop intuition about interface dynamics. The experiments highlight the essential role of initial interfacial perturbations in determining the dynamic response of the interface. The experiments produce dramatic, unexpected effects when initial perturbations are controlled and inhibited. These activities help students to develop insight about unstable interfaces that can be applied to analogous problems in materials science and engineering. The lessons examine ``Rayleigh-Taylor instability,`` an interfacial instability that occurs when a higher-density fluid is above a lower-density fluid.

Benjamin, R.F. [Los Alamos National Lab., NM (United States). Dynamic Experimentation Div.



Learning to show you're listening  

Microsoft Academic Search

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 sequence which enables learners to acquire a basic Arabic back-channel

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



Show Them You Really Want the Job  

ERIC Educational Resources Information Center

Showing that one really "wants" the job entails more than just really wanting the job. An interview is part Broadway casting call, part intellectual dating game, part personality test, and part, well, job interview. When there are 300 applicants for a position, many of them will "fit" the required (and even the preferred) skills listed in the job…

Perlmutter, David D.



The Morning Show at WLES-TV.  

ERIC Educational Resources Information Center

|Describes the production and programing of daily quarter-hour television shows by different groups of students at Laurel (Maryland) Elementary School, guided by the library media specialist who started them five years ago. The video experience has improved students' reading, writing, and math skills, as well as behavior. (MF)|

Blondell, Beverley



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



Olaparib shows promise in multiple tumor types.  


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



The Last Great American Picture Show  

Microsoft Academic Search

The Last Great American Picture Show brings together essays by scholars and writers who chart the changing evaluations of the American cinema of the 1970s, sometimes referred to as the decade of the lost generation, but now more and more recognized as the first New Hollywood, without which the cinema of Francis Coppola, Steven Spielberg, Robert Zemeckis, Tim Burton or

Thomas Elsaesser; Noel King; Alexander Horwath



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



Children's Art Show: An Educational Family Experience  

ERIC Educational Resources Information Center

|In a time of seemingly rampant budget cuts in the arts in school systems throughout the country, a children's art show reaps many rewards. It can strengthen family-school relationships and community ties and stimulate questions and comments about the benefits of art and its significance in the development of young children. In this photo essay of…

Bakerlis, Julienne



Show Them You Really Want the Job  

ERIC Educational Resources Information Center

|Showing that one really "wants" the job entails more than just really wanting the job. An interview is part Broadway casting call, part intellectual dating game, part personality test, and part, well, job interview. When there are 300 applicants for a position, many of them will "fit" the required (and even the preferred) skills listed in the job…

Perlmutter, David D.



A Fasciclin-Like Arabinogalactan-Protein (FLA) Mutant of Arabidopsis thaliana, fla1, Shows Defects in Shoot Regeneration  

PubMed Central

Background The fasciclin-like arabinogalactan-proteins (FLAs) are an enigmatic class of 21 members within the larger family of arabinogalactan-proteins (AGPs) in Arabidopsis thaliana. Located at the cell surface, in the cell wall/plasma membrane, they are implicated in many developmental roles yet their function remains largely undefined. Fasciclin (FAS) domains are putative cell-adhesion domains found in extracellular matrix proteins of organisms from all kingdoms, but the juxtaposition of FAS domains with highly glycosylated AGP domains is unique to plants. Recent studies have started to elucidate the role of FLAs in Arabidopsis development. FLAs containing a single FAS domain are important for the integrity and elasticity of the plant cell wall matrix (FLA11 and FLA12) and FLA3 is involved in microspore development. FLA4/SOS5 with two FAS domains and two AGP domains has a role in maintaining proper cell expansion under salt stressed conditions. The role of other FLAs remains to be uncovered. Method/Principal Findings Here we describe the characterisation of a T-DNA insertion mutant in the FLA1 gene (At5g55730). Under standard growth conditions fla1-1 mutants have no obvious phenotype. Based on gene expression studies, a putative role for FLA1 in callus induction was investigated and revealed that fla1-1 has a reduced ability to regenerate shoots in an in vitro shoot-induction assay. Analysis of FLA1p:GUS reporter lines show that FLA1 is expressed in several tissues including stomata, trichomes, the vasculature of leaves, the primary root tip and in lateral roots near the junction of the primary root. Conclusion The results of the developmental expression of FLA1 and characterisation of the fla1 mutant support a role for FLA1 in the early events of lateral root development and shoot development in tissue culture, prior to cell-type specification.

Johnson, Kim L.; Kibble, Natalie A. J.; Bacic, Antony; Schultz, Carolyn J.



Kepler Systems That Show Multiple Transiting Objects  

NASA Astrophysics Data System (ADS)

Exoplanetary systems that have multiple transiting planets provide unique and important insight into the formation, evolution, and dynamics of exoplanetary systems. Kepler has announced the discovery of a confirmed planetary system with multiple transiting planets (Kepler 9, Holman et al. 2010) as well as several candidate planetary systems that show multiple transiting objects (Steffen et al. 2010). Kepler 9 shows deviations from a constant period due to the ongoing dynamical interactions between the confirmed planets. From these transit timing variations (TTV) one can measure the planetary masses from the photometric data alone. The presence of several systems with multiple transiting candidates from the first quarter of data indicate that Kepler should continue to find systems with multiple transiting planets. Such systems will provide important, general information about the histories of planetary systems.

Steffen, Jason H.; Fabrycky, D. C.; Ford, E. B.; Holman, M. J.; Lissauer, J. J.; Ragozzine, D.; Welsh, W. F.; Kepler Science Team



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.


Hungry females show stronger mating preferences  

Microsoft Academic Search

Female mating decisions that are based on condition-dependent traits, such as male nutritional state, may be associated with a female's own condition. In the swordtail fish, Xiphophorus birchmanni, females prefer the chemical cues of well-fed males to cues of food-deprived males. Here we show that this preference is significantly stronger in females when they were experimentally food deprived than when

Heidi S. Fisher; Gil G. Rosenthal



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 impacted each other. The continuing collision of the Antennae galaxies, located about 62 million light years from Earth, has triggered the formation of millions of stars in clouds of dusts and gas.

Jim Wilson



Do dogs ( Canis familiaris ) show contagious yawning?  

Microsoft Academic Search

We report an experimental investigation into whether domesticated dogs display contagious yawning. Fifteen dogs were shown\\u000a video clips of (1) humans and (2) dogs displaying yawns and open-mouth expressions (not yawns) to investigate whether dogs\\u000a showed contagious yawning to either of these social stimuli. Only one dog performed significantly more yawns during or shortly\\u000a after viewing yawning videos than to

Aimee L. Harr; Valerie R. Gilbert; Kimberley A. Phillips



An invariant aspartic acid in the DNA glycosylase domain of DEMETER is necessary for transcriptional activation of the imprinted MEDEA gene.  


Helix-hairpin-helix DNA glycosylases are typically small proteins that initiate repair of DNA by excising damaged or mispaired bases. An invariant aspartic acid in the active site is involved in catalyzing the excision reaction. Replacement of this critical residue with an asparagine severely reduces catalytic activity but preserves enzyme stability and structure. The Arabidopsis DEMETER (DME) gene encodes a large 1,729-aa polypeptide with a 200-aa DNA glycosylase domain. DME is expressed primarily in the central cell of the female gametophyte. DME activates maternal allele expression of the imprinted MEDEA (MEA) gene in the central cell and is required for seed viability. We mutated the invariant aspartic acid at position 1304 in DME to asparagine (D1304N) to determine whether the catalytic activity of the DNA glycosylase domain is required for DME function in vivo. Transgenes expressing wild-type DME in the central cell rescue seed abortion caused by a mutation in the endogenous DME gene and activate maternal MEA:GFP transcription. However, transgenes expressing the D1304N mutant DME do not rescue seed abortion or activate maternal MEA:GFP transcription. Whereas ectopic expression of the wild-type DME polypeptide in pollen is sufficient to activate ectopic paternal MEA and MEA:GUS expression, equivalent expression of the D1304N mutant DME in pollen failed to do so. These results show that the conserved aspartic acid residue is necessary for DME to function in vivo and suggest that an active DNA glycosylase domain, normally associated with DNA repair, promotes gene transcription that is essential for gene imprinting. PMID:15128940

Choi, Yeonhee; Harada, John J; Goldberg, Robert B; Fischer, Robert L



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.



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



Clathrin triskelia show evidence of molecular flexibility.  


The clathrin triskelion, which is a three-legged pinwheel-shaped heteropolymer, is a major component in the protein coats of certain post-Golgi and endocytic vesicles. At low pH, or at physiological pH in the presence of assembly proteins, triskelia will self-assemble to form a closed clathrin cage, or "basket". Recent static light scattering and dynamic light scattering studies of triskelia in solution showed that an individual triskelion has an intrinsic pucker similar to, but differing from, that inferred from a high resolution cryoEM structure of a triskelion in a clathrin basket. We extend the earlier solution studies by performing small-angle neutron scattering (SANS) experiments on isolated triskelia, allowing us to examine a higher q range than that probed by static light scattering. Results of the SANS measurements are consistent with the light scattering measurements, but show a shoulder in the scattering function at intermediate q values (0.016 A(-1)), just beyond the Guinier regime. This feature can be accounted for by Brownian dynamics simulations based on flexible bead-spring models of a triskelion, which generate time-averaged scattering functions. Calculated scattering profiles are in good agreement with the experimental SANS profiles when the persistence length of the assumed semiflexible triskelion is close to that previously estimated from the analysis of electron micrographs. PMID:18502808

Ferguson, Matthew L; Prasad, Kondury; Boukari, Hacene; Sackett, Dan L; Krueger, Susan; Lafer, Eileen M; Nossal, Ralph



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



Hospital survey shows improvements in patient experience.  


Hospitals are improving the inpatient care experience. A government survey that measures patients' experiences with a range of issues from staff responsiveness to hospital cleanliness-the Hospital Consumer Assessment of Healthcare Providers and Systems survey-is showing modest but meaningful gains. Using data from the surveys reported in March 2008 and March 2009, we present the first comprehensive national assessment of changes in patients' experiences with inpatient care since public reporting of the results began. We found improvements in all measures of patient experience, except doctors' communication. These improvements were fairly uniform across hospitals. The largest increases were in measures related to staff responsiveness and the discharge information that patients received. PMID:21041749

Elliott, Marc N; Lehrman, William G; Goldstein, Elizabeth H; Giordano, Laura A; Beckett, Megan K; Cohea, Christopher W; Cleary, Paul D



Microbiological and environmental issues in show caves.  


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



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.



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



Cloning and characterization of a calcium dependent protein kinase gene associated with cotton fiber development  

Microsoft Academic Search

The gene GhCPK1 encoding a calcium dependent protein kinase was identified from cotton. Transcripts of GhCPK1 accumulated primarily in the elongating fiber, and Arabidopsis plants transformed with GhCPK1 promoter-GUS construct exhibited GUS activity mainly in the developing trichomes, roots, young leaves and sepals. In the bombarded onion\\u000a epidermal cells, GhCPK1-GFP fusion proteins showed a subcellular distribution in the plasma membrane.

Quan-Sheng Huang; Hai-Yun Wang; Peng Gao; Guo-Ying Wang; Gui-Xian Xia



T-DNA tagging in Brassica napus as an efficient tool for the isolation of new promoters for selectable marker genes  

Microsoft Academic Search

A simple strategy to identify and isolate new promoters suitable for driving the expression of selectable marker genes is described. By employing a Brassica napus hypocotyl transformation protocol and a promoterless gus::nptII tagging construct, a series of 20 kanamycin-resistant tagged lines was produced. Most of the regenerated plants showed hardly any GUS activity in leaf, stem and root tissues. However,

Jacob Bade; Emiel van Grinsven; Jerome Custers; Sietske Hoekstra; Anne Ponstein



Do men and women show love differently in marriage?  


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



Characterization of a PttRPS18 promoter active in the vascular cambium region of hybrid aspen.  


By screening 273 hybrid aspen plants transformed with a luciferase-based promoter trap T-DNA vector, one plant was found in which the reporter gene (luxF2) was activated only in cells of the cambial region, i.e., vascular cambium, phloem and differentiating xylem. Southern blot analysis revealed that this plant denoted #24 had a single T-DNA insert. The chromosomal regions flanking the T-DNA were cloned by plasmid rescue. A 757 bp DNA fragment, originating from the rescued plasmid and covering the genomic region immediately upstream from the right-border sequence of the T-DNA, was used as a probe to isolate the corresponding chromosomal region from a wild-type hybrid aspen genomic library. A hybrid aspen small ribosomal protein gene, PttRPS18, was then isolated. By screening a wt cambial region-specific cDNA library, two cDNA clones encoding a putative 152 amino acid PttRPS18 protein were isolated. Comparison of the DNA sequence immediately flanking the T-DNA insert in #24 with the corresponding wild-type sequence showed that only a minor deletion occurred during the T-DNA integration. Northern analysis revealed that the PttRPS18 gene was expressed mainly in the cambial region. By RT-PCR and DNA sequencing analysis, the exact structures of the PttRPS18 and luxF2 transcripts were determined. Finally, the hybrid aspen PttRPS18 promoter was fused to the uidA reporter gene and transformed into hybrid aspen plants. Histochemical analysis of GUS activity showed that the PttRPS18 promoter was expressed in the cambial region in the same manner as the luciferase reporter gene in the initial screening. PMID:12856939

Johansson, Ann-Marie; Wang, Chongying; Stenberg, Anneli; Hertzberg, Magnus; Little, C H Anthony; Olsson, Olof



Transgenic apple (Malus x domestica) shoot showing low browning potential.  


Transgenic apple shoots were prepared from leaf disks by using Agrobacterium tumefaciens carrying the kanamycin (KM) resistance gene and antisense polyphenol oxidase (PPO) DNA. Four transgenic apple lines that grew on the medium containing 50 microgram/mL KM were obtained. They contained the KM resistance gene and grew stably on the medium for >3 years. Two transgenic shoot lines containing antisense PPO DNA in which PPO activity was repressed showed a lower browning potential than a control shoot. PMID:11087467

Murata, M; Haruta, M; Murai, N; Tanikawa, N; Nishimura, M; Homma, S; Itoh, Y



Activation of camalexin biosynthesis in Arabidopsis thaliana in response to perception of bacterial lipopolysaccharides: a gene-to-metabolite study.  


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



Designed oligomers of cyanovirin-N show enhanced HIV neutralization.  


Cyanovirin-N (CV-N) is a small, cyanobacterial lectin that neutralizes many enveloped viruses, including human immunodeficiency virus type I (HIV-1). This antiviral activity is attributed to two homologous carbohydrate binding sites that specifically bind high mannose glycosylation present on envelope glycoproteins such as HIV-1 gp120. We created obligate CV-N oligomers to determine whether increasing the number of binding sites has an effect on viral neutralization. A tandem repeat of two CV-N molecules (CVN(2)) increased HIV-1 neutralization activity by up to 18-fold compared to wild-type CV-N. In addition, the CVN(2) variants showed extensive cross-clade reactivity and were often more potent than broadly neutralizing anti-HIV antibodies. The improvement in activity and broad cross-strain HIV neutralization exhibited by these molecules holds promise for the future therapeutic utility of these and other engineered CV-N variants. PMID:21799112

Keeffe, Jennifer R; Gnanapragasam, Priyanthi N P; Gillespie, Sarah K; Yong, John; Bjorkman, Pamela J; Mayo, Stephen L



Designed oligomers of cyanovirin-N show enhanced HIV neutralization  

PubMed Central

Cyanovirin-N (CV-N) is a small, cyanobacterial lectin that neutralizes many enveloped viruses, including human immunodeficiency virus type I (HIV-1). This antiviral activity is attributed to two homologous carbohydrate binding sites that specifically bind high mannose glycosylation present on envelope glycoproteins such as HIV-1 gp120. We created obligate CV-N oligomers to determine whether increasing the number of binding sites has an effect on viral neutralization. A tandem repeat of two CV-N molecules (CVN2) increased HIV-1 neutralization activity by up to 18-fold compared to wild-type CV-N. In addition, the CVN2 variants showed extensive cross-clade reactivity and were often more potent than broadly neutralizing anti-HIV antibodies. The improvement in activity and broad cross-strain HIV neutralization exhibited by these molecules holds promise for the future therapeutic utility of these and other engineered CV-N variants.

Keeffe, Jennifer R.; Gnanapragasam, Priyanthi N. P.; Gillespie, Sarah K.; Yong, John; Bjorkman, Pamela J.; Mayo, Stephen L.




ERIC Educational Resources Information Center

|Presents three activities: (1) investigating succession in a schoolground; (2) investigating oak galls; and (3) making sun prints (photographs made without camera or darkroom). Each activity includes a list of materials needed and procedures used. (JN)|

Clearing: Nature and Learning in the Pacific Northwest, 1984



The postmitotic Saccharomyces cerevisiae after spaceflight showed higher viability  

NASA Astrophysics Data System (ADS)

The budding yeast Saccharomyces cerevisiae has been proposed as an ideal model organism for clarifying the biological effects caused by spaceflight conditions. The postmitotic S. cerevisiae cells onboard Practice eight recoverable satellite were subjected to spaceflight for 15 days. After recovery, the viability, the glycogen content, the activities of carbohydrate metabolism enzymes, the DNA content and the lipid peroxidation level in yeast cells were analyzed. The viability of the postmitotic yeast cells after spaceflight showed a three-fold increase as compared with that of the ground control cells. Compared to the ground control cells, the lipid peroxidation level in the spaceflight yeast cells markedly decreased. The spaceflight yeast cells also showed an increase in G2/M cell population and a decrease in Sub-G1 cell population. The glycogen content and the activities of hexokinase and succinate dehydrogenase significantly decreased in the yeast cells after spaceflight. In contrast, the activity of malate dehydrogenase showed an obvious increase after spaceflight. These results suggested that microgravity or spaceflight could promote the survival of postmitotic S. cerevisiae cells through regulating carbohydrate metabolism, ROS level and cell cycle progression.

Yi, Zong-Chun; Li, Xiao-Fei; Wang, Yan; Wang, Jie; Sun, Yan; Zhuang, Feng-Yuan



D-Ser-containing humanin shows promotion of fibril formation.  


Humanin (HN), a peptide of 24 amino acid residues, suppresses the neuronal cell death that is induced by the gene products of Alzheimer's disease. HN contains two Ser residues at positions 7 and 14. Because the proportion of D-Ser isomerized from L-Ser in proteins appears to increase as cellular organs age, we explored the structural effects of the isomerization of each Ser residue in HN. By using a thioflavin-T assay to detect fibril formation, we found that an HN derivative that contained two isomerized D-Ser residues had a greater tendency to form fibrils than did wild-type HN or HNs containing single D-Ser residues. A previous report showed that HN containing two D-Ser residues exerts neuroprotective activity. Our data, therefore, suggest that the fibril formation by HN that contains two D-Ser residues may promote HN neuroprotective activity. PMID:21735222

Hayashi, Kanehiro; Sasabe, Jumpei; Chiba, Tomohiro; Aiso, Sadakazu; Utsunomiya-Tate, Naoko



Electricity show and related educational programming. Final report  

SciTech Connect

The on-site version of Balance of Power reached a total audience of 21,331 between November 10, 1994 and January 31, 1996; in addition, the Physics on Wheels van offered exhibits and programs to an additional 30,000 students in the 1995-1996 school year. The program provided a groundbreaking new approach to informal science education, combining a dynamic demonstration with an intensely interactive game show. Between the on-site programming and the Physics on Wheels van programs, 51,331 students were impacted by the activities, exhibits and energy-conservation message of Balance of Power.




Prkcz null mice show normal learning and memory  

PubMed Central

Protein kinase M ? (PKM?) is a constitutively active form of atypical PKC that is exclusively expressed in the brain and implicated in the maintenance of long-term memory1–9. Most studies that support a role for PKM? in memory maintenance have used pharmacological PKM? inhibitors such as the myristoylated zeta inhibitory peptide (ZIP) or chelerythrine. Here, we used a genetic approach and targeted exon 9 of the Prkcz gene to generate mice that lack both protein kinase C ? (PKC?) and PKM? (Prkcz?/? mice). Prkcz?/? mice showed normal behavior in a cage environment and in baseline tests of motor function and sensory perception, but displayed reduced anxiety-like behavior. Surprisingly, they did not show deficits in learning or memory in tests of cued fear conditioning, novel object recognition, object location recognition, conditioned place preference (CPP) for cocaine, or motor learning, when compared with wild-type littermates. ZIP injection into the nucleus accumbens (NAc) reduced expression of cocaine CPP in Prkcz?/? mice. In vitro, ZIP and scrambled ZIP inhibited PKM?, PKC? and PKC? with similar Ki values. Chelerythrine was a weak inhibitor of PKM? (Ki = 76 µM). Our findings show that absence of PKM? does not impair learning and memory in mice, and that ZIP can erase reward memory even when PKM? is not present.

Lee, Anna M.; Kanter, Benjamin R.; Wang, Dan; Lim, Jana P.; Zou, Mimi E.; Qiu, Chichen; McMahon, Tom; Dadgar, Jahan; Fischbach-Weiss, Sarah C.; Messing, Robert O.



Student Demographics, Extracurricular Participation and Safety Education of Students Participating in the 2003 Houston Livestock Show and Rodeo Agricultural Mechanics Project Show  

Microsoft Academic Search

The Houston Livestock Show and Rodeo Agricultural Mechanics Project Show is the largest show of it kind in Texas, and perhaps the largest in the nation. This extracurricular activity provides students and agricultural education programs an opportunity to display skills developed in agricultural mechanics laboratories by exhibiting projects constructed entirely by students. Projects such as gooseneck trailers, bulk feeders, cattle

Doug Ulrich; Dwayne Pavelock; Joe Muller; Billy Harrell


Cloning and expression of beta-glucuronidase from Lactobacillus brevis in E. coli and application in the bioconversion of baicalin and wogonoside.  


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



Leucine Aminopeptidase RNAs, Proteins, and Activities Increase in Response to Water Deficit, Salinity, and the Wound Signals Systemin, Methyl Jasmonate, and Abscisic Acid1  

PubMed Central

LapA RNAs, proteins, and activities increased in response to systemin, methyl jasmonate, abscisic acid (ABA), ethylene, water deficit, and salinity in tomato (Lycopersicon esculentum). Salicylic acid inhibited wound-induced increases of LapA RNAs. Experiments using the ABA-deficient flacca mutant indicated that ABA was essential for wound and systemin induction of LapA, and ABA and systemin acted synergistically to induce LapA gene expression. In contrast, pin2 (proteinase inhibitor 2) was not dependent on exogenous ABA. Whereas both LapA and le4 (L. esculentum dehydrin) were up-regulated by increases in ABA, salinity, and water deficit, only LapA was regulated by octadecanoid pathway signals. Comparison of LapA expression with that of the PR-1 (pathogenesis-related 1) and GluB (basic ?-1,3-glucanase) genes indicated that these PR protein genes were modulated by a systemin-independent jasmonic acid-signaling pathway. These studies showed that at least four signaling pathways were utilized during tomato wound and defense responses. Analysis of the expression of a LapA1:GUS gene in transgenic plants indicated that the LapA1 promoter was active during floral and fruit development and was used during vegetative growth only in response to wounding, Pseudomonas syringae pv tomato infection, or wound signals. This comprehensive understanding of the regulation of LapA genes indicated that this regulatory program is distinct from the wound-induced pin2, ABA-responsive le4, and PR protein genes.

Chao, Wun S.; Gu, Yong-Qiang; Pautot, Veronique; Bray, Elizabeth A.; Walling, Linda L.



Secondary wall NAC binding element (SNBE), a key cis-acting element required for target gene activation by secondary wall NAC master switches  

PubMed Central

The biosynthesis of secondary walls in vascular plants requires the coordinated regulation of a suite of biosynthetic genes, and this coordination has recently been shown to be executed by the secondary wall NAC (SWN)-mediated transcriptional network. In Arabidopsis, five SWNs, including SND1, NST1/2 and VND6/7, function as master transcriptional switches to activate their common targets and consequently the secondary wall biosynthetic program. A recent report by Zhong et al.1 revealed that SWNs bind to a common cis-acting element, namely secondary wall NAC binding element (SNBE), which is composed of an imperfect palindromic 19-bp consensus sequence, (T/A)NN(C/T)(T/C/G)TNNNNNNNA(A/C)GN(A/C/T) (A/T). Genome-wide analysis of direct targets of SWNs showed that SWNs directly activate the expression of not only many transcription factors but also a battery of genes involved in secondary wall biosynthesis, cell wall modification and programmed cell death, the promoters of which all contain multiple SNBE sites. The functional significance of the SNBE sites is further substantiated by our current in planta expression study demonstrating that representative SNBE sequences from several SWN direct target promoters are sufficient to drive the expression of the GUS reporter gene in secondary wall-forming cells. The identification of the SWN DNA binding element (SNBE) and the SWN direct targets marks an important step forward toward the dissection of the transcriptional network regulating the biosynthesis of secondary walls, the most abundant biomass produced by land plants.

McCarthy, Ryan L; Zhong, Ruiqin



New Inspiring Planetarium Show Introduces ALMA to the Public  

NASA Astrophysics Data System (ADS)

As part of a wide range of education and public outreach activities for the International Year of Astronomy 2009 (IYA2009), ESO, together with the Association of French Language Planetariums (APLF), has produced a 30-minute planetarium show, In Search of our Cosmic Origins. It is centred on the global ground-based astronomical Atacama Large Millimeter/submillimeter Array (ALMA) project and represents a unique chance for planetariums to be associated with the IYA2009. ESO PR Photo 09a/09 Logo of the ALMA Planetarium Show ESO PR Photo 09b/09 Galileo's first observations with a telescope ESO PR Photo 09c/09 The ALMA Observatory ESO PR Photo 09d/09 The Milky Way band ESO PR Video 09a/09 Trailer in English ALMA is the leading telescope for observing the cool Universe -- the relic radiation of the Big Bang, and the molecular gas and dust that constitute the building blocks of stars, planetary systems, galaxies and life itself. It is currently being built in the extremely arid environment of the Chajnantor plateau, at 5000 metres altitude in the Chilean Andes, and will start scientific observations around 2011. ALMA, the largest current astronomical project, is a revolutionary telescope, comprising a state-of-the-art array of 66 giant 12-metre and 7-metre diameter antennas observing at millimetre and submillimetre wavelengths. In Search of our Cosmic Origins highlights the unprecedented window on the Universe that this facility will open for astronomers. "The show gives viewers a fascinating tour of the highest observatory on Earth, and takes them from there out into our Milky Way, and beyond," says Douglas Pierce-Price, the ALMA Public Information Officer at ESO. Edited by world fulldome experts Mirage3D, the emphasis of the new planetarium show is on the incomparable scientific adventure of the ALMA project. A young female astronomer guides the audience through a story that includes unique animations and footage, leading the viewer from the first observations by Galileo, 400 years ago, to the world of modern astronomy, moving from the visible wavelength domain to explore the millimetre-wave view of the Universe, and leaving light-polluted cities for unique settings in some of the highest and driest places on Earth. "The fascinating topic, the breathtaking ESO astronomical images, the amazing 3D computer animations, and the very clever use of the music, all make this a really inspiring show," says Agnčs Acker, President of the APLF. In search of our Cosmic Origins is available in three different formats: fulldome video, classical with video windows, and classical with slides. Fulldome video shows immerse the audience in a true 360-degree projected computer-generated virtual environment. The ALMA planetarium show is currently available in French and English. Several other language versions are in preparation: German, Italian, Spanish and Chilean Spanish, while further languages are planned: Danish, Dutch, Greek, Japanese, Portuguese and Brazilian Portuguese. The show will be available to all planetariums worldwide for a small fee, depending on the type and the size of the planetarium, to cover basic costs. The media are invited to attend, and see firsthand, the official screening during the European Film Festival, between 24 and 26 April 2009 in Espinho, Portugal. For media accreditation, please go to A set of educational materials is also being prepared and will be finished in late April. To learn more about the show, please go to



Structured learning of human interactions in TV shows.  


The objective of this work is recognition and spatiotemporal localization of two-person interactions in video. Our approach is person-centric. As a first stage we track all upper bodies and heads in a video using a tracking-by-detection approach that combines detections with KLT tracking and clique partitioning, together with occlusion detection, to yield robust person tracks. We develop local descriptors of activity based on the head orientation (estimated using a set of pose-specific classifiers) and the local spatiotemporal region around them, together with global descriptors that encode the relative positions of people as a function of interaction type. Learning and inference on the model uses a structured output SVM which combines the local and global descriptors in a principled manner. Inference using the model yields information about which pairs of people are interacting, their interaction class, and their head orientation (which is also treated as a variable, enabling mistakes in the classifier to be corrected using global context). We show that inference can be carried out with polynomial complexity in the number of people, and describe an efficient algorithm for this. The method is evaluated on a new dataset comprising 300 video clips acquired from 23 different TV shows and on the benchmark UT--Interaction dataset. PMID:23079467

Patron-Perez, Alonso; Marszalek, Marcin; Reid, Ian; Zisserman, Andrew



Caveolin-2-deficient mice show increased sensitivity to endotoxemia  

PubMed Central

Caveolin proteins are structural components of caveolae and are involved in the regulation of many biological processes. Recent studies have shown that caveolin-1 modulates inflammatory responses and is important for sepsis development. In the present study, we show that caveolin-1 and caveolin-2 have opposite roles in lipopolysaccharide (LPS)-induced sepsis using caveolin-deficient (Cav-1-/- and Cav-2-/-) mice for each of these proteins. While Cav-1-/- mice displayed delayed mortality following challenge with LPS, Cav-2-/- mice were more sensitive to LPS compared to wild-type (WT). With Cav-2-/- mice, this effect was associated with increased intestinal injury and increased intestinal permeability. This negative outcome was also correlated with enhanced expression of iNOS in intestinal epithelial cells, and enhanced production of nitric oxide (NO). By contrast, Cav-1-/- mice demonstrated a decrease in iNOS expression with decreased NO production, but no alteration in intestinal permeability. The differential expression of iNOS was associated with a significant increase in STAT-1 activation in these mice. Intestinal cells of Cav-2-/- mice showed increased phosphorylation of STAT-1 at tyrosine 701 compared to wild-type. However, Cav-1-/- mice-derived intestinal cells showed decreased levels of phosphorylation of STAT-1 at tyrosine 701. Since caveolin-2 is almost completely absent in Cav-1-/- mice, we conclude that it is not just the absence of caveolin-2 that is responsible for the observed effects, but that the balance between caveolin-1 and caveolin-2 is important for iNOS expression and ultimately for sepsis outcome.

de Almeida, Cecilia J; Witkiewicz, Agnieszka K; Jasmin, Jean-Francois; Tanowitz, Herbert B; Sotgia, Federica; Frank, Philippe G



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



Analysis of multiple bone responses to graded strains above functional levels, and to disuse, in mice in vivo show that the human Lrp5 G171V High Bone Mass mutation increases the osteogenic response to loading but that lack of Lrp5 activity reduces it  

PubMed Central

Introduction To investigate the role of the low-density lipoprotein receptor-related protein 5 (Lrp5) in bones' responses to loading, we analysed changes in multiple measures of bone architecture in tibias subjected to loading or disuse in male and female mice with the Lrp5 loss of function mutation (Lrp5?/?) or heterozygous for the Lrp5 G171V High Bone Mass (HBM) mutation (Lrp5HBM+). Materials and methods The right tibias of these 17 week old male and female mice and their Wild Type (WT) littermates were subjected to short periods of loading three days a week for two weeks. Each tibia was loaded for 40 cycles, to produce peak strains at the midshaft within the low, medium or high physiological range (~ 1500, 2400 and 3000 microstrain, respectively). In similar groups of mice the right sciatic nerve was severed causing disuse of the right tibia for 3 weeks. Data from microCT of loaded, neurectomised and contra-lateral control tibias were analysed to quantify changes in the cortical and cancellous regions of the bone in the absence of functional strains and in response to graded strains in addition to those derived from function. Results and conclusion Male WT+/+ controls showed significant strain:response curves for cortical area and trabecular thickness, but Lrp5?/? mice showed no detectable strain:response in those same outcomes. Female mice of either WT+/+ or Lrp5?/? genotype did not show significant strain:response curves for cortical or trabecular parameters, the one exception being Tb.Th in Lrp5?/? mice. Since female WT+/+ mice did not respond to loading in a significant dose:responsive manner, the similar lack of responsiveness of the Lrp5?/? females could not be ascribed to their Lrp5 status. Cortical bone loss associated with disuse showed no differences between Lrp5?/? mice and WT+/+ controls, but in cancellous bone of both male and females of these mice, there was a greater loss than in WT+/+ controls. In contrast, the tibias of male and female mice heterozygous for the Lrp5 G171V HBM mutation showed greater osteogenic responsiveness to loading and less bone loss associated with disuse than their WTHBM? controls. These data indicate that the presence of the Lrp5 G171V HBM mutation is associated with an increased osteogenic response to loading but support only a marginal gender-related role for normal Lrp5 function in this loading-related response.

Saxon, Leanne K.; Jackson, Brendan F.; Sugiyama, Toshihiro; Lanyon, Lance E.; Price, Joanna S.



Tests show organic clay stabilizers prevent permeability loss  

SciTech Connect

Water sensitivity of sandstones can result in rapid and severe permeability impairment if freshwater displaces saline connate water from pore spaces of formation rock. Reduced permeability can decrease the production potential of a well. Laboratory test on sandstone cores show that organic clay stabilizers (OCS) are as effective at preventing permeability reduction from freshwater exposure as commonly used potassium chloride (KCl). Although organic clay stabilizers are widely used, only limited laboratory testing has confirmed their effectiveness at preventing permeability reduction attributable to a formation's water sensitivity. Existing test results on one class of OCS indicated satisfactory performance as a KCl replacement; however, the test procedure included only short cores and the simulated brine used for testing was sodium chloride (NaCl) at 5 % concentration. The recent work included a series of short-core and long-core laboratory tests made at various salinity levels and OCS concentrations. The OCS tested is a high-concentration, non-surface active, ammonium salt in an aqueous fluid.

Behenna, R. (Halliburton Energy Services, Duncan, OK (United States))



Crayfish brain-protocerebrum and retina show serotonergic functional relationship.  


The results from various studies have indicated possible functional relationships between crayfish electroretinogram (ERG) rhythmic amplitude changes and the serotonergic pathways projecting from the central brain through the optic neuropils to the eye, but to date, this functional interaction has not been proven. Here, in a set of experiments using an isolated eyestalk-brain preparation, we investigated whether there is a circadian input from the brain to retina that regulates this rhythm. We sought to determine whether the protocerebral bridge (PB) stimulation affects the ERG amplitude in accordance with the zeitgeber time (ZT) and whether 5-HT modulates the associate input. Our results showed that photic stimulation of retina produced changes in both the amplitude and the frequency of spontaneous electrical activity in the protocerebral neuropils. In addition, electrical stimulation of the medial protocerebrum, particularly the PB, produced statistically significant changes in the ERG that depended on both the time of day and the level of serotonin. This suggests that pathways between retina and PB seem to be serotonergic. PMID:21911207

Valdés-Fuentes, Marlen; Prieto-Sagredo, Julio; Fanjul-Moles, María Luisa



Gem and Mineral Shows as Geologic Teaching Opportunities.  

ERIC Educational Resources Information Center

|Gem and mineral shows are excellent nontraditional opportunities for community education and outreach by geology teachers. Discusses initial club contacts, displays, shows, and the advantages of show participation to academic geologists. (CW)|

Cordua, William Sinclair



Analysis of mechanisms regulating expression of the ver-1 gene, involved in aflatoxin biosynthesis.  


Previous studies have shown that ver-1A encodes an enzyme which is directly involved in the conversion of versicolorin A to demethylsterigmatocystin during aflatoxin B1 (AFB1) biosynthesis in the filamentous fungus Aspergillus parasiticus. In this study, two different tools were utilized to study the regulation of ver-1A expression at the level of transcription and protein accumulation. First, a ver-1A cDNA was expressed in Escherichia coli with the vector pMAL-c2. The resulting maltose-binding protein-Ver-1A fusion protein was purified and used to generate polyclonal antibodies. Western blot analyses showed that these antibodies specifically recognized the Ver-1 protein (approximately 28 kDa) in cell extracts of Aspergillus parasiticus SU1. Second, a GUS (uidA; encodes beta-glucuronidase) reporter system was developed by fusing the ver-1A promoter and transcription terminator to the GUS gene. Reporter constructs were transformed into A. parasiticus, resulting in a single copy of the ver-1A-GUS reporter integrated adjacent to the wild-type ver-1A gene (3' end) in the chromosome. Western blot analysis, Northern hybridization analysis, and a GUS activity assay were used to analyze transformants. The timing of appearance and pattern of accumulation of GUS transcript and GUS protein in transformants were consistent with the timing of appearance and pattern of accumulation of ver-1 transcript and Ver-1 protein. These data suggested that the GUS gene was under the same regulatory control as the wild-type ver-1 gene and confirmed that transcriptional regulation plays an important role in ver-1A expression. Integration of the ver-1A-GUS reporter construct at the niaD locus resulted in 500-fold-lower GUS activity, but the temporal pattern of accumulation of GUS activity was not affected. Therefore, chromosomal location can play a role in determining the level of gene expression in A. parasiticus and should be an important consideration when analyzing promoter function in this organism. PMID:9055421

Liang, S H; Wu, T S; Lee, R; Chu, F S; Linz, J E



site plan, floor plan, southeast and east elevations, detail showing ...  

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

site plan, floor plan, southeast and east elevations, detail showing original front entrance, interior detail showing fireplace in elevation - Neiman House, 1930 Providence Road, Charlotte, Mecklenburg County, NC


Daytime television talk shows: Guests, content and interactions  

Microsoft Academic Search

Strident controversy over the often?bizarre subject matter of daytime television talk shows motivated this content analysis of 11 shows with the highest Nielsen ratings in 1994–95. A sample of 10 episodes of each of the shows was videotaped and subjected to a systematic analysis of the shows’ guests, topics of discussion and interactions. Results indicate that this genre of program

Bradley S. Greenberg; John L. Sherry; Rick W. Busselle; Lynn Rampoldi Hnilo; Sandi W. Smith



"The George Lopez Show": The Same Old Hispano?  

ERIC Educational Resources Information Center

|"The George Lopez Show" is the first successful television show with a Latino in a leading role that features Hispanic material since Freddie Prinze's thirty-year-old sitcom, "Chico and the Man." This study seeks to assess how Latinos are presented on "The George Lopez Show." A content analysis reveals that the show perpetuates some of the…

Markert, John



Reducing appointment no-shows: going from theory to practice.  


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



Ulvan, a Sulfated Polysaccharide from Green Algae, Activates Plant Immunity through the Jasmonic Acid Signaling Pathway  

PubMed Central

The industrial use of elicitors as alternative tools for disease control needs the identification of abundant sources of them. We report on an elicitor obtained from the green algae Ulva spp. A fraction containing most exclusively the sulfated polysaccharide known as ulvan-induced expression of a GUS gene placed under the control of a lipoxygenase gene promoter. Gene expression profiling was performed upon ulvan treatments on Medicago truncatula and compared to phytohormone effects. Ulvan induced a gene expression signature similar to that observed upon methyl jasmonate treatment (MeJA). Involvement of jasmonic acid (JA) in ulvan response was confirmed by detecting induction of protease inhibitory activity and by hormonal profiling of JA, salicylic acid (SA) and abscisic acid (ABA). Ulvan activity on the hormonal pathway was further consolidated by using Arabidopsis hormonal mutants. Altogether, our results demonstrate that green algae are a potential reservoir of ulvan elicitor which acts through the JA pathway.

Jaulneau, Valerie; Lafitte, Claude; Jacquet, Christophe; Fournier, Sylvie; Salamagne, Sylvie; Briand, Xavier; Esquerre-Tugaye, Marie-Therese; Dumas, Bernard