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Sample records for 35s enhancer sequence

  1. Unconventional P-35S sequence identified in genetically modified maize.

    Al-Hmoud, Nisreen; Al-Husseini, Nawar; Ibrahim-Alobaide, Mohammed A; Kübler, Eric; Farfoura, Mahmoud; Alobydi, Hytham; Al-Rousan, Hiyam

    2014-01-01

    The Cauliflower Mosaic Virus 35S promoter sequence, CaMV P-35S, is one of several commonly used genetic targets to detect genetically modified maize and is found in most GMOs. In this research we report the finding of an alternative P-35S sequence and its incidence in GM maize marketed in Jordan. The primer pair normally used to amplify a 123 bp DNA fragment of the CaMV P-35S promoter in GMOs also amplified a previously undetected alternative sequence of CaMV P-35S in GM maize samples which we term V3. The amplified V3 sequence comprises 386 base pairs and was not found in the standard wild-type maize, MON810 and MON 863 GM maize. The identified GM maize samples carrying the V3 sequence were found free of CaMV when compared with CaMV infected brown mustard sample. The data of sequence alignment analysis of the V3 genetic element showed 90% similarity with the matching P-35S sequence of the cauliflower mosaic virus isolate CabbB-JI and 99% similarity with matching P-35S sequences found in several binary plant vectors, of which the binary vector locus JQ693018 is one example. The current study showed an increase of 44% in the incidence of the identified 386 bp sequence in GM maize sold in Jordan's markets during the period 2009 and 2012. PMID:24495911

  2. Electrochemical biosensor for the detection of cauliflower mosaic virus 35 S gene sequences using lead sulfide nanoparticles as oligonucleotide labels.

    Sun, Wei; Zhong, Jianghua; Qin, Peng; Jiao, Kui

    2008-06-15

    Lead sulfide (PbS) nanoparticles were synthesized in aqueous solution and used as oligonucleotide labels for electrochemical detection of the 35 S promoter from cauliflower mosaic virus (CaMV) sequence. The PbS nanoparticles were modified with mercaptoacetic acid and could easily be linked with CaMV 35 S oligonucleotide probe. Target DNA sequences were covalently linked on a mercaptoacetic acid self-assembled gold electrode, and DNA hybridization of target DNA with probe DNA was completed on the electrode surface. PbS nanoparticles anchored on the hybrids were dissolved in the solution by oxidation of HNO3 and detected using a sensitive differential pulse anodic stripping voltammetric method. The detection results can be used to monitor the hybridization reaction. The CaMV 35 S target sequence was satisfactorily detected with the detection limit as 4.38 x 10(-12)mol/L (3sigma). The established method extends nanoparticle-labeled electrochemical DNA analysis to specific sequences from genetically modified organisms with higher sensitivity and selectivity. PMID:18381196

  3. Simultaneous electrochemical DNA hybridization assay for PAT and FMV 35S gene sequence using quantum dots as labels

    Jiang Hua Zhong; Peng Qin; Wei Sun; Kui Jiao

    2008-01-01

    An electrochemical method for the simultaneous detection of two different DNA sequenees from PAT and FMV 35S gene sequence using CdS and PbS quantum dots (QDs)as labels was described.The QDs were readily functionalized with oligonucleotides as electrochemical DNA probes and selectively hybridized to the complementary sequences immobilized on the microplate.The QDs anchored on the hybrids were dissolved in the solution by the oxidation of HNO3 and further detected by a sensitive differential pulse anodic stripping voltammetric method(DPASV).The DPASV signals of the oxidation of Cd2+ and Pb2+ ions present in the solution were difierent and reflected the identity of corresponding ssDNA targets sequences.

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

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

    2007-08-01

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

  5. Main: CTRMCAMV35S [PLACE

    Full Text Available CTRMCAMV35S S000460 27-March-2004 (last modified) kehi CT-rich motif (inverted GAGA...) found in a 60-nucleotide region (S1) downstream of the transcription start site of the CaMV 35S RNA; Can e...nhance gene expression; Inverted GAGA; See also S000405, S000427 (GAGA); (TC)4T; CaMV 35S; enhancer; Cauliflower mosaic virus (CaMV) TCTCTCTCT ...

  6. CNS depressants accelerate the dissociation of /sup 35/S-TBPS binding and GABA enhances their displacing potencies

    Maksay, G.; Ticku, M.K.

    1988-01-01

    The specific binding of /sup 35/S-t-butylbicyclophosphorothionate (TBPS) was studied in synaptosomal membranes of rat cerebral cortex. The displacing potencies of eleven CNS depressants and three convulsants were determined in the presence of 1 /sup +/M GABA and 10 nM R 5135. GABA enhanced the displacing potencies of depressants of most diverse chemical structures: diaryltriazine (LY 81067), pyrazolopyridine (etazolate), cinnamide, glutarimide, 2,3-benzodiazepine (tofizopam) and alcohol derivatives, barbiturates, (+)etomidate, methaqualone and meprobamate. In contrast, the IC/sub 50/ values of convulsants (picrotoxinin, pentetrazol and the barbiturate enantiomer S(+)MPPB) were not significantly affected. The depressants accelerated either basal or GABA-augmented dissociation of /sup 35/-TBPS mainly by increasing the contribution of its rapid first phase.

  7. Comparison of in situ DNA hybridization protocols using 35S-labeled and biotin-labeled probes in detection of human papillomavirus DNA sequences

    This paper describes a modified in situ DNA hybridization technique with 35S-labeled HPV DNA probes (Technique II) compared against the method (Technique I) routinely used by the authors, as well as against a new technique utilizing biotin-labeled probes (Technique III) the latter employed a 12-hour hybridization with biotin-labeled HPV DNA probes and reaction visualization with avidin, biotinylated alkaline phosphatase, and a substrate kit (Vector). The modifications in Technique II included omission of the 0.2 N HCI wash, increased proteinase-Κ concentration, elevated denaturation temperature (110-1200C), as well as replacement of poly-D-lysine as a slide-coating medium by Kodak Photo-Flo 200. In a series of cervical and penile human papillomavirus (HPV) lesions tested, identical HPV DNA types were discovered by Techniques I and II, whereas three specimens remained negative with Technique III

  8. Preparation of 35S labelled thiosemicarbazone

    A 35S labelled thiosemicarbazone is prepared, on a millimole scale by reacting labelled thiocyanate with hydrazine sulfate in ethanolic medium. The hydrazine thiocyanate so formed is then condensed with aldehyde to form the thiosemicarbazone

  9. Selection of DNA clones with enhancer sequences.

    Asoh, S; Lee-Kwon, W; Mouradian, M M; Nirenberg, M

    1994-01-01

    A method is described for selection of DNA clones that contain enhancer sequences that activate gene expression. An Escherichia coli-rodent cell shuttle vector, pPyE0, was used that contains polyoma viral DNA without the polyoma enhancer region. Replication of pPyE0 DNA in mouse cells is markedly reduced due to deletion of the polyoma enhancer region. Insertion of mouse genomic DNA fragments that contain putative enhancer sequences into pPyE0 adjacent to the polyoma origin of replication rest...

  10. Biosynthetic preparation of 35-S labelled methionine

    High specific activity methionine with sulfur-35 was prepared in our laboratory by growing Baker's yeast cells, in a medium containing 35S-sulfate. L-S35 methionine was prepared from the acid hydrolyzate of the proteins by chromatography on whatman paper. The specific activity was determined using o-phtaladehyde as a fluorophore to form a fluorescent complex. The specific activity was found to be usually greater than 800 Ci/mmol. (Author)

  11. Enhanced Dynamic Algorithm of Genome Sequence Alignments

    Arabi E. keshk

    2014-05-01

    Full Text Available The merging of biology and computer science has created a new field called computational biology that explore the capacities of computers to gain knowledge from biological data, bioinformatics. Computational biology is rooted in life sciences as well as computers, information sciences, and technologies. The main problem in computational biology is sequence alignment that is a way of arranging the sequences of DNA, RNA or protein to identify the region of similarity and relationship between sequences. This paper introduces an enhancement of dynamic algorithm of genome sequence alignment, which called EDAGSA. It is filling the three main diagonals without filling the entire matrix by the unused data. It gets the optimal solution with decreasing the execution time and therefore the performance is increased. To illustrate the effectiveness of optimizing the performance of the proposed algorithm, it is compared with the traditional methods such as Needleman-Wunsch, Smith-Waterman and longest common subsequence algorithms. Also, database is implemented for using the algorithm in multi-sequence alignments for searching the optimal sequence that matches the given sequence.

  12. Soil 35S Transformation and Availability to Plants

    ZHOUWEI; PANJIARONG

    1999-01-01

    Sulfur transformation in 3 soils maintained in a closed incubation system and its availability to plants were investigated using carrier-free 35S-SO42- and 35S-labeled ryegrass straw.For carrier-free Na235SO4 treatment,78%,70%and 64% of 35S applied were found in Ca(H2PO4)2-extractale S fraction,4%,5% and 7% in slowly soluble inorganic S,11%,15%and 18%in C-O-S,5%,7%,and 6% in C-bonded S,and 5%,7% and 6% in unidentified organic S120 days after incubation in black soil,cinnamon soil and chestnut soil,respectively.Most of 35S uptake by plants came from extractable 35SO42-,and little from C-O-35S and C-bonded 35S,In the treatment with 35S-labeled straw,51%,46%and 36% of 35S incorporated were found in Ca(H2PO4)2-extractable S fraction,7%,6% and 7% in slowly soluble inorganic S,13%,15%and 18% in C-O-S,8%,8%and 6% in C-bonded S,and 18%,25%and 35% in unidentified organic S at the end of incubation in above-mentoned three soils,respectively.Higher availability of C-O-35S,C-bonded 35S and unidentified organic 35S from 35S-labeled straw was observed in 35S-labeled straw treatment compared to carrier-free Na235SO4 treatment.

  13. The effect of ethanol on 35-S-TBPS binding to mouse brain membranes in the presence of chloride

    The effect of in vitro and in vivo administration of ethanol on the binding of 35S-t-butyl-bicyclophosphorothionate (35S-TBPS) to cortical brain membranes of C57B1 mice was investigated using KCl (100 mM) containing assay media. The in vitro addition of ethanol produced a dose-dependent inhibition of basal 35S-TBPS binding. In the presence of chloride ions, GABA and pentobarbital had a biphasic action (stimulation followed by inhibition) on 35S-TBPS binding, whereas diazepam only stimulated the binding. Ethanol reduced the stimulatory effects of GABA and pentobarbital in a dose-dependent manner, but had no effect on the enhancement of 35S-TBPS binding produced by diazepam. 35S-TBPS binding to cortical brain membranes was inhibited by the putative Cl- channel blocking agent DIDS. This inhibitory action of DIDS was significantly, and dose-dependently reduced by ethanol (≤ 100 mM ethanol). Chronic ethanol ingestion in vivo, which produced tolerance to and physical dependence on ethanol in the animals, did not alter the stimulatory and inhibitory effects of GABA and pentobarbital on 35S-TBPS binding. The enhancement of 35S-TBPS binding produced by diazepam was slightly, but significantly, enhanced in brain membranes from animals which had undergone 24 hours of ethanol withdrawal. Chronic ethanol treatment did not change the potency of picrotoxin and of the peripheral BDZ-receptor ligand RO 5-4864 to competitively inhibit 35S-TBPS binding. Our results suggest that in vitro addition of ethanol alters the activity of the activity of the GABA benzodiazepine (BDZ) receptor complex. Although there was no change in basal 35S-TBPS binding following chronic in vivo ethanol administration, our curent data suggest that chronic ethanol ingestion may cause specific changes of the GABA BDZ receptor proteins, in this study revealed as an altered modulation of 35S-TBPS binding by diazepam. (author)

  14. Separation of 35S-sulfate on neutral aluminium oxide

    The results of 35S-sulfate chromatography on neutral aluminium oxide (γ-Al2O3) are presented. Possibility of quantitative adsorption of 35S-sulfate on Al2O3 from acidic or neutral potassium chloride solutions of high concentration is shown. Dynamic adsorption capacity of neutral aluminium oxide with respect to sulfate from weakly acidic potassium chloride solution equals near 10 μmol/ml of adsorbent. Optimal parameters for chromatographic isolation of 35S-sulfate without carrier from irradiated KCl target are determined. (author)

  15. Development of a general method for detection and quantification of the P35S promoter based on assessment of existing methods.

    Wu, Yuhua; Wang, Yulei; Li, Jun; Li, Wei; Zhang, Li; Li, Yunjing; Li, Xiaofei; Li, Jun; Zhu, Li; Wu, Gang

    2014-01-01

    The Cauliflower mosaic virus (CaMV) 35S promoter (P35S) is a commonly used target for detection of genetically modified organisms (GMOs). There are currently 24 reported detection methods, targeting different regions of the P35S promoter. Initial assessment revealed that due to the absence of primer binding sites in the P35S sequence, 19 of the 24 reported methods failed to detect P35S in MON88913 cotton, and the other two methods could only be applied to certain GMOs. The rest three reported methods were not suitable for measurement of P35S in some testing events, because SNPs in binding sites of the primer/probe would result in abnormal amplification plots and poor linear regression parameters. In this study, we discovered a conserved region in the P35S sequence through sequencing of P35S promoters from multiple transgenic events, and developed new qualitative and quantitative detection systems targeting this conserved region. The qualitative PCR could detect the P35S promoter in 23 unique GMO events with high specificity and sensitivity. The quantitative method was suitable for measurement of P35S promoter, exhibiting good agreement between the amount of template and Ct values for each testing event. This study provides a general P35S screening method, with greater coverage than existing methods. PMID:25483893

  16. Enhanced Dynamic Algorithm of Genome Sequence Alignments

    Arabi E. keshk

    2014-01-01

    The merging of biology and computer science has created a new field called computational biology that explore the capacities of computers to gain knowledge from biological data, bioinformatics. Computational biology is rooted in life sciences as well as computers, information sciences, and technologies. The main problem in computational biology is sequence alignment that is a way of arranging the sequences of DNA, RNA or protein to identify the region of similarity and relationship between se...

  17. Comparison of fluorographic methods for the detection of 35S-labeled proteins in polyacrylamide gels

    Eight different methods of fluorographic enhancement of sensitivity to 35S decay after gel electrophoresis were compared. Using Kodak X-Omat AR X-ray film, we found that some fluors were about equivalent to 2,5-diphenyloxazole/dimethyl sulfoxide embedding, whereas several other fluors were not quite as effective, but still were significantly more sensitive than control autoradiography. The most sensitive procedures can yield a detectable darkening of film with less than 1 dpm/mm2 of 35S after a 1-week exposure

  18. Reservoir enhancements and production technology sequencing

    Dusseault, M.B. [Waterloo Univ., ON (Canada). Dept. of Earth Sciences, Porous Media Research Inst.

    2006-07-01

    Criteria used to select production technologies for reservoirs with high levels of sand production were discussed. The paper examined a range of thermal recovery methods including cyclic steam stimulation (CSS) and steam assisted gravity drainage (SAGD). VAPEX, hybrid steam, and in situ combustion processes were reviewed. The review indicated that reservoir processes that involve high levels of sand production, high pressures, and high temperatures can cause changes in porosity and breach or shear barriers and shale beds within reservoirs. The changes can be beneficial to technologies designed to reduce flow path lengths in gravity drainage processes. Reservoir scale changes associated with cold heavy oil production with sand (CHOPS) and steam injection technologies were also discussed. It was concluded that careful sequencing of technologies can have a significant impact on recovery rates. 12 refs., 13 figs.

  19. Enhanced learning of natural visual sequences in newborn chicks.

    Wood, Justin N; Prasad, Aditya; Goldman, Jason G; Wood, Samantha M W

    2016-07-01

    To what extent are newborn brains designed to operate over natural visual input? To address this question, we used a high-throughput controlled-rearing method to examine whether newborn chicks (Gallus gallus) show enhanced learning of natural visual sequences at the onset of vision. We took the same set of images and grouped them into either natural sequences (i.e., sequences showing different viewpoints of the same real-world object) or unnatural sequences (i.e., sequences showing different images of different real-world objects). When raised in virtual worlds containing natural sequences, newborn chicks developed the ability to recognize familiar images of objects. Conversely, when raised in virtual worlds containing unnatural sequences, newborn chicks' object recognition abilities were severely impaired. In fact, the majority of the chicks raised with the unnatural sequences failed to recognize familiar images of objects despite acquiring over 100 h of visual experience with those images. Thus, newborn chicks show enhanced learning of natural visual sequences at the onset of vision. These results indicate that newborn brains are designed to operate over natural visual input. PMID:27079969

  20. Synthetic heparinoids labelled with 125I and 35S

    Sederel, L.C.; Kolar, Z.; Does, van der, Leen; Bantjes, A.

    1982-01-01

    The labelling of a water-soluble synthetic polyelectrolyte, having anticoagulant activity, has been studied. The polyelectrolyte is derived from cis-1,4-polyisoprene and contains N-sulfate and carboxylate groups. [125I]-Iodination of the polyelectrolyte, using the Chloramine-T method and an electrolytic method, resulted in a [125I]-labelled polyelectrolyte from which release of the label occurred. Resulfation of a partially desulfated polyelectrolyte with a [35S]-sulfur trioxide trimethylamin...

  1. Production of some inorganic forms of 35S at the Boris Kidric Institute

    Several inorganic forms of 35S are used as starting material for some chemical syntheses of labelled organic compounds. The choice depends on the desired characteristics of the labelled compound. Preparation of elementary sulphur 35S, sodium sulphide-35S, ferrous sulphide-35S, barium sulphate-35S, barium sulphide-35S, is described. The characteristics of the products are given along with the determination methods. The products are obtained with high specific activities: elementary sulphur-35S, 1 - 5 Ci/gS; sodium sulphide-35S 1 - 5 Ci/gS; ferrous sulphide-35S, 1 - 5 Ci/gS; barium sulphate-35S, 5 - 15 Ci/gS; barium sulphide-35S, 1 - 5 Ci/gS. (author)

  2. Enhanced Landmine Detection from Low Resolution IR Image Sequences

    Wang, Tiesheng; Gu, Irene Yu-Hua; Tjahjadi, Tardi

    We deal with the problem of landmine field detection using low-resolution infrared (IR) image sequences measured from airborne or vehicle-borne passive IR cameras. The proposed scheme contains two parts: a) employ a multi-scale detector, i.e., a special type of isotropic bandpass filters, to detect landmine candidates in each frame; b) enhance landmine detection through seeking maximum consensus of corresponding landmine candidates over image frames. Experiments were conducted on several IR image sequences measured from airborne and vehicle-borne cameras, where some results are included. As shown in our experiments, the landmine signatures have been significantly enhanced using the proposed scheme, and automatic detection results are reasonably good. These methods can therefore be applied to assisting humanitarian demining work for landmine field detection.

  3. Privacy-Enhanced Methods for Comparing Compressed DNA Sequences

    Eppstein, David; Baldi, Pierre

    2011-01-01

    In this paper, we study methods for improving the efficiency and privacy of compressed DNA sequence comparison computations, under various querying scenarios. For instance, one scenario involves a querier, Bob, who wants to test if his DNA string, $Q$, is close to a DNA string, $Y$, owned by a data owner, Alice, but Bob does not want to reveal $Q$ to Alice and Alice is willing to reveal $Y$ to Bob \\emph{only if} it is close to $Q$. We describe a privacy-enhanced method for comparing two compressed DNA sequences, which can be used to achieve the goals of such a scenario. Our method involves a reduction to set differencing, and we describe a privacy-enhanced protocol for set differencing that achieves absolute privacy for Bob (in the information theoretic sense), and a quantifiable degree of privacy protection for Alice. One of the important features of our protocols, which makes them ideally suited to privacy-enhanced DNA sequence comparison problems, is that the communication complexity of our solutions is pr...

  4. An experimental study of targeting therapy with 35S-SZ39 against glioma

    Objective: To prepare a pure β-emitting immunoradiotherapeutics agent 35S-MAb SZ39, and validate its special therapeutic efficacy against glioma. Methods: MAb SZ39 was labelled with 35S using a carbodiimide method. Using 35S-nIgG, 35S + MAb SZ39 and sustained 35S as control agents, and human brain glioma cell line SHG-44 as target cell, the injury rate and 50% inhibitory concentration of 35S-MAb SZ39 were evaluated with MTT method. 35S-MAb SZ39 and its control agent 35S-nIgG or PBS were i.p. injected into glioma-bearing nude mice. The tumor inhibitory rate (I) was determined according to the formula: I = [1-(TV35S-MAb/TVPBS)] x 100% (TV: tumor volume). Flowcytometry was used to analyse the cell cycle of glioma after treatment. Results: 35S-MAb SZ39 had a strong cytotoxic effect to glioma cells with 4.2-fold and 4.0-fold more toxic than 35S-nIgG and 35S + MAb SZ39, as strong as the sustained 35S control group. Tumor growth blocking for one week was obtained with 103.6 MBq 35S-MAb SZ39 treatment. The inhibitory rate was 50% 26 days after 35S-MAb SZ39 administration. DNA synthesis of glioma cells was inhibited, cells were accumulated in S period and the road to G1 period was blocked. There was a trend of cell cycle synchronization. No obvious toxicity was found on bone marrow while 35S-MAb SZ39 made the glioma growth block. Conclusions: 35S-MAb SZ39 has a strong selective injurious effect on glioma and is of good prospect to be an immunoradiotherapeutics agent

  5. Recombinase Polymerase Amplification (RPA) of CaMV-35S Promoter and nos Terminator for Rapid Detection of Genetically Modified Crops

    Chao Xu; Liang Li; Wujun Jin; Yusong Wan

    2014-01-01

    Recombinase polymerase amplification (RPA) is a novel isothermal DNA amplification and detection technology that enables the amplification of DNA within 30 min at a constant temperature of 37–42 °C by simulating in vivo DNA recombination. In this study, based on the regulatory sequence of the cauliflower mosaic virus 35S (CaMV-35S) promoter and the Agrobacterium tumefaciens nopaline synthase gene (nos) terminator, which are widely incorporated in genetically modified (GM) crops, we designed t...

  6. Real-time imaging of 35S-sulfate uptake in a rape seed plant

    We present real-time images of 35S-sulfate uptake in a rapeseed plant visualized by the system we developed. In the leaves of rapeseed plants, 35S accumulated in higher amounts and more rapidly in the more developed leaves. This real-time imaging system can be used to visualize the movement of both 35S and 32P in the same plant. In the pods of rapeseed, images of 35S show that 35S accumulated mostly in the terminal parts; on the other hand 32P, when applied as 32P-phosphoric acid, accumulated in the middle part of the pods. (orig.)

  7. Possible consequences of the overlap between the CaMV 35S promoter regions in plant transformation vectors used and the viral gene VI in transgenic plants.

    Podevin, Nancy; du Jardin, Patrick

    2012-01-01

    Multiple variants of the Cauliflower mosaic virus 35S promoter (P35S) are used to drive the expression of transgenes in genetically modified plants, for both research purposes and commercial applications. The genetic organization of the densely packed genome of this virus results in sequence overlap between P35S and viral gene VI, encoding the multifunctional P6 protein. The present paper investigates whether introduction of P35S variants by genetic transformation is likely to result in the expression of functional domains of the P6 protein and in potential impacts in transgenic plants. A bioinformatic analysis was performed to assess the safety for human and animal health of putative translation products of gene VI overlapping P35S. No relevant similarity was identified between the putative peptides and known allergens and toxins, using different databases. From a literature study it became clear that long variants of the P35S do contain an open reading frame, when expressed, might result in unintended phenotypic changes. A flowchart is proposed to evaluate possible unintended effects in plant transformants, based on the DNA sequence actually introduced and on the plant phenotype, taking into account the known effects of ectopically expressed P6 domains in model plants. PMID:22892689

  8. The percutaneous absorption of 35S-acetyl-L-methionine and L-serine in rabbit

    The authors had reported that L-cysteine probably was formed from acetyl-L-methionine and L-serine through cystathionine pathway by the skin enzyme of rabbit, and the solution composed of acetyl-L-methionine and L-serine exhibited the effectiveness to the hair growth in rabbit. This report shows that, by the application of 35S-acetyl-L-methionine and L-serine to the skin of rabbit and in vitro analysis of the metabolites of 35S-compounds, 35S-acetyl-L-methionine was absorbed into the hair tissues for many hours, and half 35S-L-cystine was formed in vitro and in vivo. When total amount of 35S in the hair was measured, the radiochemical activities were clearly shown as almost 35S-L-cystine. (auth.)

  9. Enhanced secretion of adhesive recognition sequence containing hirudin III mutein in E. coli.

    Tan, Shuhua; Wu, Wutong; Li, Xiangyu; Cui, Li; Li, Bing; Ruan, Qiping

    2007-05-01

    It has been previously shown that Escherichia coli L-asparaginase II (L-ASP) signal peptide is capable of being utilized to direct extracellular secretion of hirudin III (HV3) in shake flask. In this study HV3 muteins R33G34D35(S36)-HV3 were generated by introduction of adhesive recognition sequence RGD(S) into the non-functional region of HV3. The resultant recombinants were cultivated on 30 l bioreactor scale using L-ASP signal peptide expression system and the optimized fed-batch cultivation was well established. After cultivation for approximately 11 h the secreted product accumulated up to approximately 1 g l(-1), which means 17-fold increase in productivity compared to initial expression in shake flask. N-terminal analysis, pI measurement, and MALDI mass spectral analysis on mutein R33G34D35S36-HV3 confirmed the authenticity of the product. Compared to wild-type HV3 and R33G34D35HV3, the mutein R33G34D35S36-HV3 exhibits the improved pharmacological activity. Collectively, a novel secretion strategy using L-ASP signal peptide for the rapid, efficient and cost-effective production of HV3 mutein possessing improved pharmacological activity on bioreactor scale has been well established. Using this expression system downstream processing becomes very simple because secreted product is mature, soluble, active, and without N-terminal extension of Met, which is quite critical for most therapeutic protein to reduce the side effect in clinic use. Thus, it provides a promising alternative for extracellular production of other difficult-to-express protein for biopharmaceutical use. PMID:17827531

  10. An enhanced algorithm for multiple sequence alignment of protein sequences using genetic algorithm

    Kumar, Manish

    2015-01-01

    One of the most fundamental operations in biological sequence analysis is multiple sequence alignment (MSA). The basic of multiple sequence alignment problems is to determine the most biologically plausible alignments of protein or DNA sequences. In this paper, an alignment method using genetic algorithm for multiple sequence alignment has been proposed. Two different genetic operators mainly crossover and mutation were defined and implemented with the proposed method in order to know the pop...

  11. Development and Validation of a P-35S, T-nos, T-35S and P-FMV Tetraplex Real-time PCR Screening Method to Detect Regulatory Genes of Genetically Modified Organisms in Food.

    Eugster, Albert; Murmann, Petra; Kaenzig, Andre; Breitenmoser, Alda

    2014-10-01

    In routine analysis screening methods based on real-time PCR (polymerase chain reaction) are most commonly used for the detection of genetically modified (GM) plant material in food and feed. Screening tests are based on sequences frequently used for GM development, allowing the detection of a large number of GMOs (genetically modified organisms). Here, we describe the development and validation of a tetraplex real-time PCR screening assay comprising detection systems for the regulatory genes Cauliflower Mosaic Virus 35S promoter, Agrobacterium tumefaciens nos terminator, Cauliflower Mosaic Virus 35S terminator and Figwort Mosaic Virus 34S promoter. Three of the four primer and probe combinations have already been published elsewhere, whereas primers and probe for the 35S terminator have been developed in-house. Adjustment of primer and probe concentrations revealed a high PCR sensitivity with insignificant physical cross-talk between the four detection channels. The sensitivity of each PCR-system is sufficient to detect a GMO concentration as low as 0.05% of the containing respective element. The specificity of the described tetraplex is high when tested on DNA from GM maize, soy, rapeseed and tomato. We also demonstrate the robustness of the system by inter-laboratory tests. In conclusion, this method provides a sensitive and reliable screening procedure for the detection of the most frequently used regulatory elements present in GM crops either authorised or unauthorised for food. PMID:25437161

  12. Real-time imaging of {sup 35}S-sulfate uptake in a rape seed plant

    Nakanishi, T.M.; Yamawaki, M.; Ishibashi, H.; Tanoi, K. [Tokyo Univ. (Japan). Lab. of Radioisotope Plant Physiology

    2011-07-01

    We present real-time images of {sup 35}S-sulfate uptake in a rapeseed plant visualized by the system we developed. In the leaves of rapeseed plants, {sup 35}S accumulated in higher amounts and more rapidly in the more developed leaves. This real-time imaging system can be used to visualize the movement of both {sup 35}S and {sup 32}P in the same plant. In the pods of rapeseed, images of {sup 35}S show that {sup 35}S accumulated mostly in the terminal parts; on the other hand {sup 32}P, when applied as {sup 32}P-phosphoric acid, accumulated in the middle part of the pods. (orig.)

  13. Enhanced MR imaging sequencing and calibration with a personal computer

    This paper reports on a commercial MR imaging system to permit all combination of parameter values that the machine can support the facilitate implementation of experimental sequences. Personal computers (PCs) were connected to Toshiba 0.35-T MR imaging systems by means of special file-transfer software. PC programs generated control sequences dynamically. Calibration was performed using numeric calculation of phase moments from a multiparameter model of gradient coil performance. An associated data base allowed new sequence types to be introduced without programming or calibration. Options included velocity nulling, cardiac multiphase imaging, and presaturation of arbitrary volumes

  14. An enhanced algorithm for multiple sequence alignment of protein sequences using genetic algorithm.

    Kumar, Manish

    2015-01-01

    One of the most fundamental operations in biological sequence analysis is multiple sequence alignment (MSA). The basic of multiple sequence alignment problems is to determine the most biologically plausible alignments of protein or DNA sequences. In this paper, an alignment method using genetic algorithm for multiple sequence alignment has been proposed. Two different genetic operators mainly crossover and mutation were defined and implemented with the proposed method in order to know the population evolution and quality of the sequence aligned. The proposed method is assessed with protein benchmark dataset, e.g., BALIBASE, by comparing the obtained results to those obtained with other alignment algorithms, e.g., SAGA, RBT-GA, PRRP, HMMT, SB-PIMA, CLUSTALX, CLUSTAL W, DIALIGN and PILEUP8 etc. Experiments on a wide range of data have shown that the proposed algorithm is much better (it terms of score) than previously proposed algorithms in its ability to achieve high alignment quality. PMID:27065770

  15. Polyacrylamide gel electrophoresis and subspeciation of total cell proteins from multi-antibiotic-resistant skin diphtheroids labelled with [35S]methionine or [35S]thioATP and of coagulase negative staphylococci labelled with [35S]methionine

    The authors demonstrate that [35S]methionine labelling of proteins followed by PAGE can be used to distinguish biochemically similar, multi-antibiotic-resistant skin diphtheroids; thirty-one isolates fell into four subgroups. The method also distinguished ten separate electrophoretypes of coagulase-negative staphylococci which correlated approximately with eight biotypes; the fifty-one isolates of biotypes SII, the commonest clinical isolate, were electrophoretically identical, thus suggesting that they are, indeed, members of a single subgroup. The authors also report a novel method of radiolabelling the phosphoproteins using [35S]thioATP and have demonstrated the method using the above diphtheroids. These were again distinguished into the same four subgroups although the patterns of phosphoproteins were qualitatively and quantitatively different from those of the proteins. This thioATP labelling method should have wide application also. 14 refs.; 7 figs.; 4 tabs

  16. Autoradiographic visualization in comparison with the incorporation of 35S-methionine by various tissue protein

    The purpose of the present study was to observe the incorporation level of 35S-methionine by various tissue protein in organism. By the use of the macro- and micro-autoradiographic technique, the incorporation of 35S-methionine by the tissues has been utilized as an index of tissue protein synthesis. Further experiments showed that 35S-methionine was dominantly incorporated in the liver, kidney and spleen. It indicated that a strong protein metabolism produced in these tissues. In spite of the important physiological function of the heart, lung and skeletal muscle, the protein metabolism in those tissues was in a low level

  17. Gonadotropin hyperstimulation influences the 35S-methionine metabolism of mouse preimplantation embryos.

    Wetzels, A M; Artz, M T; Goverde, H J; Bastiaans, B A; Hamilton, C J; Rolland, R

    1995-11-01

    The effects of gonadotropin stimulation on mouse embryo uptake and incorporation of 35S-methionine were studied. We found that the uptake of 35S-methionine was reduced in embryos of stimulated females in both the two-cell and the blastocyst developmental stage. The incorporation of 35S-methionine into protein was not statistically significantly different between the embryos of stimulated and those of unstimulated females. Qualitatively, protein synthesis was equal in both groups as determined with one-dimensional SDS-PAGE. The results are discussed and we conclude that mouse embryo viability in vivo is decreased by ovarian stimulation. PMID:8624434

  18. Accumulation of [35S]taurine in peripheral layers of the olfactory bulb

    Accumulation of [35S]taurine in the laminae of the olfactory bulb of the adult cat, rat, mouse and rabbit was examined autoradiographically. [35S]Taurine was administered either i.p. or i.v. and olfactory bulbs were excised 24 h post-injection. High concentrations of [35S]taurine were restricted to the olfactory nerve and glomerular layers of the olfactory bulb in all species examined. Olfactory neurons are continuously renewed and the results obtained suggest that taurine may have an important role in olfactory receptor axons. (Auth.)

  19. How much contrast is enough? Dependence of enhancement on field strength and MR pulse sequence

    The overwhelming majority of published studies defining the clinical utility of gadolinium administration for neuroimaging have been performed at high field using conventional spin-echo imaging. Concerning the issue of field strength, several investigations have now shown that for a given dose of contrast, enhancement is less apparent at low field than at high field. Concerning the issue of pulse sequence, there is now convincing clinical and experimental evidence that all T1-weighted sequences are not equal in demonstrating contrast enhancement. Specifically, T1-weighted spoiled gradient-echo sequences do not show the same degree of visually apparent contrast enhancement compared to conventional spin-echo sequences. The use of magnetization transfer techniques which demonstrate areas of enhancement unseen on conventional pulse sequences is also addressed. (orig.)

  20. CPMG sequences with enhanced sensitivity to chemical exchange

    Improved relaxation-compensated Carr-Purcell-Meiboom-Gill pulse sequences are reported for studying chemical exchange of backbone 15N nuclei. In contrast to the original methods [J. P. Loria, M. Rance, and A. G. Palmer, J. Am. Chem. Soc.121, 2331-2332 (1999)], phenomenological relaxation rate constants obtained using the new sequences do not contain contributions from 1H-1H dipole-dipole interactions. Consequently, detection and quantification of chemical exchange processes are facilitated because the relaxation rate constant in the limit of fast pulsing can be obtained independently from conventional 15N spin relaxation measurements. The advantages of the experiments are demonstrated using basic pancreatic trypsin inhibitor

  1. Studies on the shelf-life of L-35S methionine

    A systematic study conducted on the shelf -life of L-35S- methionine, an important radiotracer used in protein synthesis experiments is reported. Aliquots of 35S-methionine from bulk sample prepared by us were kept under chosen conditions of storage and were analysed by paper chromatography coupled with autoradiography. The radiochemical purity (RCP) of 35S-methionine at various time interval spanning over a period of about one half-life of 35S radioisotope (87.4 days) was determined. It was observed that the RCP came down only to about 89% from the original value of 96% at the end of the period of study under the chosen conditions. (author)

  2. Behavior of 32P, 35S, 36Cl and 42K in magnesium oxide

    A separation method of 32P from 35S using magnesium oxide as adsorbent of radiophosphorus is described. The behaviour of 32P and 35S, both carrier-free, on magnesium oxide, individually, in dependence of the amount of the adsorbent, of mixing time, of the pH of the loading solution and of potassium chloride concentration, is studied. The separation of the mentioned radioisotopes, using a misture of them, is also analysed. In order to apply this method to the routine production of carrier-free 35S by potassium chloride irradiation, the adsorption behaviour of the chloride and potassium on magnesium oxide using radioactive tracers of these elements, is studied. The separation of 35S from 32P is analyzed by the maximum range of β- particles in aluminum. The absorption curves are presented and compared. (Author)

  3. Sequence Comparison for Non-Enhanced MRA of the Lower Extremity Arteries at 7 Tesla

    Sören Johst; Stephan Orzada; Anja Fischer; Lena C Schäfer; Kai Nassenstein; Lale Umutlu; Lauenstein, Thomas C.; Ladd, Mark E.; Stefan Maderwald

    2014-01-01

    In this study three sequences for non-contrast-enhanced MRA of the lower extremity arteries at 7T were compared. Cardiac triggering was used with the aim to reduce signal variations in the arteries. Two fast single-shot 2D sequences, a modified Ultrafast Spoiled Gradient Echo (UGRE) sequence and a variant of the Quiescent-Interval Single-Shot (QISS) sequence were triggered via phonocardiogram and compared in volunteer examinations to a non-triggered 2D gradient echo (GRE) sequence. For image ...

  4. Protoneus-sequence: extended fluid-attenuated inversion recovery MR imaging without and with contrast enhancement

    Nasel, Christian [Division of Neuroradiology, Department of Radiology, Medical University of Vienna, Waehringerguertel 18-20, A-1090 Vienna (Austria)]. E-mail: christian.nasel@perfusion.at

    2005-08-01

    Fluid-attenuated inversion recovery imaging (=flair imaging) is widely used as primary screening sequence in various investigation protocols, due to its high lesion contrast and sensitivity in detection of parenchymatous and leptomeningeal disease. An additional increase of sensitivity for detection of lesions may be achieved by contrast-enhanced flair imaging. Based on flair imaging a dual-echo inversion recovery imaging sequence (=proton echo usage [=protoneus] - sequence) was developed, which could significantly extend the possibilities of conventional flair imaging.

  5. Multiple redundant sequence elements within the fission yeast ura4 replication origin enhancer

    Zhang Dong-Yi

    2001-01-01

    Full Text Available Abstract Background Some origins in eukaryotic chromosomes fire more frequently than others. In the fission yeast, Schizosaccharomyces pombe, the relative firing frequencies of the three origins clustered 4-8 kbp upstream of the ura4 gene are controlled by a replication enhancer - an element that stimulates nearby origins in a relatively position-and orientation-independent fashion. The important sequence motifs within this enhancer were not previously localized. Results Systematic deletion of consecutive segments of ~50, ~100 or ~150 bp within the enhancer and its adjacent core origin (ars3002 revealed that several of the ~50-bp stretches within the enhancer contribute to its function in partially redundant fashion. Other stretches within the enhancer are inhibitory. Some of the stretches within the enhancer proved to be redundant with sequences within core ars3002. Consequently the collection of sequences important for core origin function was found to depend on whether the core origin is assayed in the presence or absence of the enhancer. Some of the important sequences in the core origin and enhancer co-localize with short runs of adenines or thymines, which may serve as binding sites for the fission yeast Origin Recognition Complex (ORC. Others co-localize with matches to consensus sequences commonly found in fission yeast replication origins. Conclusions The enhancer within the ura4 origin cluster in fission yeast contains multiple sequence motifs. Many of these stimulate origin function in partially redundant fashion. Some of them resemble motifs also found in core origins. The next step is to identify the proteins that bind to these stimulatory sequences.

  6. MRI of osteomyelitis. Sensitivity and specificity of STIR sequences in comparison to contrast-enhanced T1 spin echo sequences

    Purpose: To evaluate the need for additional MR sequences including administration of Gd-DTPA after inconspicuous short-tau inversion-recovery (STIR) sequence to exclude the diagnosis of osteomyelitis. Material and Methods: 112 MR examinations of 79 patients acquired for the detection of possible osteomyelitis were analyzed retrospectively. All examinations were performed at 0.5 T including STIR, T1-weighted spin echo sequences (T1 SE) before and after application of Gd-DTPA. Additionally, 93 T2-weighted spin echo sequences were available. The examinations were analyzed by two experienced radiologists. First, the STIR sequences were studied, followed by the T1 SE images before and after administration of contrast material. Finally, the T2-weighted images were evaluated. Diagnoses were confirmed by operation (22), biopsy (10), and follow-7p (80). Results: In 53 cases osteomyelitis was diagnosed, while the remaining 59 cases suffered from another disease. The sensitivity of the STIR sequence was 100% while the specificity for osteomyelitis was 49.2%. The specificity increased to 79.7% by including T1 SE images into the analysis and reached 83.1% after considering the contrast enhanced images. T2-weighted images yielded no additional information. Conclusion: The combination of STIR and T1SE images shows a high sensitivity and specificity for osteomyelitis, thus obviating the need for any additional examinations. (orig.)

  7. Effect of anticonvulsant drugs on (35S)t-butylbicyclophosphorothionate binding in vitro and ex vivo

    Using several concentrations of eight anticonvulsant drugs in clinical use (carbamazepine, clonazepam, phenytoin, phenobarbital, ethosuximide, primidone, sodium valproate, and D,L-γ-vinyl GABA), we studied their abilities in vitro to displace (35S)t-butylbicyclophosphorothionate (35S-TBPS) from its binding site in a homogenate of rat brain. Thereafter ethosuximide (150 mg/kg), phenobarbital (30 mg/kg), clonazepam (0.3 mg/kg), or phenytoin (100 mg/kg) was injected intraperitoneally into rats for 16-20 days; and the effect of drug administration on 35S-TBPS binding was studied in the cortex and hippocampus ex vivo. Phenobarbital (100 μM, P35S-TBPS binding in vitro by 10-16%. After drug administration of phenobarbital (concentration in plasma 168 μM), the number of binding sites decreased and the binding affinity (p35S-TBPS binding in vitro at the concentration analogous to therapeutic plasma levels or ex vivo at the dose used. These results suggest that the use of phenobarbital may modulate the TBPS binding site, but the role of the present findings in the anticonvulsant action of phenobarbital needs to be further studied. (author)

  8. Studies on absorption, conversion of sulfate by rape seedling and accumulation of glucosinolate in rape seed by using 35S

    The seedlings grown in sandy culture absorbed 35SO42- from cultural solution rapidly and the 35S was incorperated into glucosinolates, amino acids and proteins. Percentage of glucosinolates-35S in extractable 35S with 70% methanol declined and those of amino acid-35S rose regularly with time after labelling. The relative content of 35S incorperated into bound constituents as well as extractable components was constant after 5 days of labelling. From two weeks after flowering, the relative content of GS-35S in pods and seeds increased linearly with time until eight weeks after flowering when all 35S extracted with 70% methanol in seeds were in form of glucosinolate. The amount of 35S-GS per pod increased linearly from the first week to the eighth week after flowering. It seems that the accumulation of GS in seeds is related to the accumulation of dry matter in seeds

  9. A new technique for selective identification and mapping of enhancers within long genomic sequences.

    Chernov, Igor; Stukacheva, Elena; Akopov, Sergey; Didych, Dmitry; Nikolaev, Lev; Sverdlov, Eugene

    2008-05-01

    We report a new experimental method of direct selection, identification, and mapping of potential enhancer sequences within extended stretches of genomic DNA. The method allows simultaneous cloning of a quantity of sequences instead of tedious screening of the separate ones, thus providing a robust and high-throughput approach to the mapping of enhancers. The selection procedure is based on the ability of such sequences to activate a minimal promoter that drives expression of a selective gene. To this end a mixture of short DNA fragments derived from the segment of interest was cloned in a retroviral vector containing the neomycin phosphotransferase II gene under control of a cytomegalovirus (CMV) minimal promoter. The pool of retroviruses obtained was used to infect HeLa cells and then to select neomycin-resistant colonies containing constructs with enhancer-like sequences. The pool of the genomic fragments was rescued by PCR and cloned, forming a library of the potential enhancers. Fifteen enhancer-like fragments were selected from 1-Mb human genome locus, and enhancer activity of 13 of them was verified in a transient transfection reporter gene assay. The sequences selected were found to be predominantly located near 5' regions of genes or within gene introns. PMID:18476831

  10. Biochemical diagnosis of mucopolysaccharidoses by estimation of intracellular 35S-sulfate accumulation

    The investigation of 35S-sulfate accumulation and of 35SO4-labelled glycosaminoglucane molecules (chase-experiments) in cultured cells was used in post- and prenatal diagnosis of mucopolysaccharidosis (MPS). Cell lines which accumulate 35S-sulfate can be differentiated by means of cross correction or measurement of enzyme activity. 34 patients with the MPS type I, II, III A, III B and VI, respectively, were diagnosed in this way. Four pregnancies at risk for MPS were prenatally monitored by the sulfate accumulation in cultured amniotic fluid cells. One fetus suffering from MPS II was found. (author)

  11. Searches for heavy neutrinos from 35S, 14C, and 63Ni beta decay

    We have searched for the effect of a neutrino of mass 17 keV/c2 in the beta decay of 35S with an apparatus incorporating a high resolution solid state detector and a super conducting solenoid. The experimental mixing probability of the 17keV neutrino is consistent with zero. The experimental sensitivity is verified by measurements with a mixed source of 35S and 14C, which artificially produces a distortion in the beta spectrum similar to that expected from the massive neutrino. Recently, we have performed similar searches in the beta decay of 14C and 63Ni. Results of these new measurements will be presented

  12. Enhancer sequences of a retroviral vector determine expression of a gene in multipotent hematopoietic progenitors and committed erythroid cells.

    Holland, C A; Anklesaria, P; Sakakeeny, M A; Greenberger, J.S.

    1987-01-01

    To analyze the transcriptional activity of retroviral enhancer sequences in hematopoietic lineages, we determined the effect of enhancer sequences on the expression of the neomycin resistance gene transferred by two retroviral vectors to primary hematopoietic lineages. We constructed the vector pFr-SV(X). The Moloney murine leukemia virus enhancer region of a vector, pZIP-SV(X), was replaced by a 380-nucleotide-long fragment containing the enhancer sequences of the Friend murine leukemia viru...

  13. The uptake of S from four different 35 S labbelled fertilizer by tea plant

    The uptake of S derived from 35-S-labelled ammonium sulphate, potasium sulphate, Kieserit (MgSO4) and Gypsum(CaSO4) of the specific activity of 0.1 mCi/g S by tea clone TRI 2025 planted in andosols was investigated. Randomized block design was used in the experiment. Fertilizers were give once at the rate of 40 kg S/ha. Counting of 35-S samples collected from the youngest and the lower leaves were done every week. Results of the experiment showed that the uptake patterns of S derived from fertilizers were the same for the youngest and the lower leaves. The activity of 35-S was clearly detected in leaves samples after one week of fertilizers application and increased continuously upto eight weeks. The total activity of 35-S in the plucked leaved derived from ammonium sulphate, potasium sulphate and Kieserite were little bit higher than from gypsum. (authors). 10 refs, 6 figs, 2 tabs

  14. Effects of 35S-dimehypo pesticide on agricultural environment and ecosystem

    Dimehypo is a nereistoxin derivative, which was developed and manufactured in China. In order of appraise its environmeatal safety comprehensively, the radioisotopic tracer technique and other methods were applied to investigate on the effects of 35S-labelled dimehypo (35S-dimehypo) on agricultural environment and ecosystem. The results revealed its low adsorptivity and high mobility, especially water-carrying mobility in soil, fair stability in soil and water, and slowness in degradation. Its main product of degradation, nereistoxin, was of lower mobility than its material compound in soil. 35S-dimehypo could enter the bodies of grass carps Ctenopharyngodon idellus along with bails or via the respiratory tracts, and could be excreted fast after the fish were removed from the contaminated source. Liquid and granulated 35S-dimehypo were fed to quail and fowls respectively, and could also be excreted rapidly in excretion and urine. Its much less distribution coefficient in caprylalcoholwater system suggested no accumulation in the adipose tissue of organism. The release of its effective composition in the granules prepared with porcelain clay or clay soil is prompt and complete

  15. The expression of foreign gene under the control of cauliflower mosaic virus 35s RNA promoter

    WangHao; BaiYongyan

    1990-01-01

    The promoter region of cauliflower mosaic virus (CaMV) 35s RNA was employed to construct an intermediate expression vector which can be used in Ti plasmid system of Agrobacterium iumefaciens.The original plasmid,which contains a polylinker between CaMV 35s RNA and its 3' termination signal in pUC18 was modified to have another antibiotic resistance marker (kanamycin resistance gene Kmr) to facilitate the selection of recombinant with Ti plasmid.Octopine synthase (ocs) structural gene was inserted into this vector downstream of CaMV 35s RNA promoter.This chimaeric gene was introduced into integrative Ti plasmid vector pGV 3850,and then transformed into Nicotiana tobaccum the chimaeric gene into tobacco cells.In both cases,the expression of ocs gene was demonstrated.The amount of octopine was much more than the nopaline synthesized by nopaline synthase (nos) gene transferred at the same time with Ti plasmid vector.This demonstrated that CaMV 35s RNA promoter is stronger in transcriptional function than the promoter of nos in tobacco cells.

  16. Determination of chemical states of sulphur 35 obtained from the 35Cl (n, p)35S

    The chemical states of sulphur-35 obtained from the 35Cl(n,p)35S reaction by the irradiation of potassium chloride without any previous treatment and with previous heating under vacuum, were determined. The influence of irradiation time and temperature after irradiation was examined. Paper electrophoresis technique was employed for the determination of the chemical states. (Author)

  17. Infection of neuroblastoma cells with Semliki Forest virus. Incorporation of 35S or 32P

    Phosphate-free medium is used for the incorporation of 32P and methionine-free medium for 35S-methionine labelling. After virus replication, the culture shows a clear CPE all of the cells appearing round and dead. Materials used are presented and experimental procedure is described

  18. Redundant sensory information does not enhance sequence learning in the serial reaction time task

    Abrahamse, Elger L.; Van der Lubbe, Rob H. J.; Verwey, Willem B.; Szumska, Izabela; Jaśkowski, Piotr

    2012-01-01

    In daily life we encounter multiple sources of sensory information at any given moment. Unknown is whether such sensory redundancy in some way affects implicit learning of a sequence of events. In the current paper we explored this issue in a serial reaction time task. Our results indicate that redundant sensory information does not enhance sequence learning when all sensory information is presented at the same location (responding to the position and/or color of the stimuli; Experiment 1), e...

  19. Redundant sensory information does not enhance sequence learning in the serial task

    Abrahamse, E.L.; Lubbe, van der, S.; Verwey, W.B.; Szumska, I.; Jaskowski, P.

    2012-01-01

    In daily life we encounter multiple sources of sensory information at any given moment. Unknown is whether such sensory redundancy in some way affects implicit learning of a sequence of events. In the current paper we explored this issue in a serial reaction time task. Our results indicate that redundant sensory information does not enhance sequence learning when all sensory information is presented at the same location (responding to the position and/or color of the stimuli; Experiment 1), e...

  20. The Role of Sleep in Motor Sequence Consolidation: Stabilization Rather Than Enhancement

    Nettersheim, Almut; Hallschmid, Manfred; Born, Jan; Diekelmann, Susanne

    2015-01-01

    Sleep supports the consolidation of motor sequence memories, yet it remains unclear whether sleep stabilizes or actually enhances motor sequence performance. Here we assessed the time course of motor memory consolidation in humans, taking early boosts in performance into account and varying the time between training and sleep. Two groups of subjects, each participating in a short wake condition and a longer sleep condition, were trained on the sequential finger-tapping task in the evening and...

  1. Distribution of /sup 35/S-sulfate within the transseptal ligament of the mouse

    Johnson, R.B.

    1989-02-01

    The distribution of 35S-sulfate-labeled macromolecules was examined within three regions of the transseptal ligament: the (1) mesial, (2) middle and (3) distal thirds. Swiss mice, 6 weeks of age, were injected with 35S-sulfate and killed after 1, 6, and 12 hours and 1, 2, 3, 4, 5, and 7 days. Silver grains and cell nuclei were counted on autoradiographs which had been counterstained by the Van Gieson method, and mean counts were analyzed statistically. Analysis of variance revealed no significant differences in mean number of cell nuclei between regions throughout the course of the experiment. 35S-sulfate was rapidly incorporated into the transseptal ligament macromolecules. Grain counts were highest 6 hours after injections: counts were highest over the middle and lowest over the mesial thirds of the ligament. The rate of grain removal was significantly higher in the middle third compared to the mesial or distal thirds (P less than 0.001) and was significantly lower in the mesial third compared to the middle or distal thirds (P less than 0.001). The half-life of labeled macromolecules was significantly greater in the mesial and distal thirds than in the middle third (P less than 0.005). The data demonstrate significantly higher rates of turnover of 35S-sulfate-labeled macromolecules in the middle region of the transseptal ligament. Since cellular density was similar throughout the transseptal ligament, higher turnover rates of 35S-sulfate-labeled macromolecules probably indicate higher rates of cellular activity in this region, possibly a result of tissue remodeling coincident to stresses generated by occlusal forces and physiologic drift of the adjacent teeth.

  2. Diagnostic accuracy of unenhanced, contrast-enhanced perfusion and angiographic MRI sequences for pulmonary embolism diagnosis: results of independent sequence readings

    To independently evaluate unenhanced, contrast-enhanced perfusion and angiographic MR sequences for pulmonary embolism (PE) diagnosis. Prospective investigation, including 274 patients who underwent perfusion, unenhanced 2D steady-state-free-precession (SSFP) and contrast-enhanced 3D angiographic MR sequences on a 1.5-T unit, in addition to CTA (CT angiography). Two independent readers evaluated each sequence independently in random order. Sensitivity, specificity, predictive values and inter-reader agreement were calculated for each sequence, excluding sequences judged inconclusive. Sensitivity was also calculated according to PE location. Contrast-enhanced angiographic sequences showed the highest sensitivity (82.9 and 89.7 %, reader 1 and reader 2, respectively), specificity (98.5 and 100 %) and agreement (kappa value 0.77). Unenhanced angiographic sequences, although less sensitive overall (68.7 and 76.4 %), were sensitive for the detection of proximal PE (92.7 and 100 %) and showed high specificity (96.1 and 99.1 %) and good agreement (kappa value 0.62). Perfusion sequences showed lower sensitivity (75.0 and 79.3 %), specificity (84.8 and 89.7 %) and agreement (kappa value 0.51), and a negative predictive value of 84.8 % at best. Compared with contrast-enhanced angiographic sequences, unenhanced sequences demonstrate lower sensitivity, except for proximal PE, but high specificity and agreement. The negative predictive value of perfusion sequences was insufficient to safely rule out PE. (orig.)

  3. Diagnostic accuracy of unenhanced, contrast-enhanced perfusion and angiographic MRI sequences for pulmonary embolism diagnosis: results of independent sequence readings

    Revel, Marie Pierre [Hopital Europeen Georges Pompidou, APHP, Departments of Radiology, Paris (France); Universite Paris Descartes Sorbonne Paris Cite, Paris (France); Hotel-Dieu, Service de Radiologie, Paris (France); Sanchez, Olivier; Meyer, Guy [Hopital Europeen Georges Pompidou, APHP, Respiratory and intensive care and, Paris (France); Universite Paris Descartes Sorbonne Paris Cite, Paris (France); INSERM Unite 765, Paris (France); Lefort, Catherine; Couchon, Sophie; Hernigou, Anne; Frija, Guy [Hopital Europeen Georges Pompidou, APHP, Departments of Radiology, Paris (France); Niarra, Ralph [Hopital Europeen Georges Pompidou, APHP, Clinical Epidemiology, Paris (France); Universite Paris Descartes Sorbonne Paris Cite, Paris (France); Chatellier, Gilles [Hopital Europeen Georges Pompidou, APHP, Clinical Epidemiology, Paris (France); Universite Paris Descartes Sorbonne Paris Cite, Paris (France); INSERM CIC-EC E4, Paris (France)

    2013-09-15

    To independently evaluate unenhanced, contrast-enhanced perfusion and angiographic MR sequences for pulmonary embolism (PE) diagnosis. Prospective investigation, including 274 patients who underwent perfusion, unenhanced 2D steady-state-free-precession (SSFP) and contrast-enhanced 3D angiographic MR sequences on a 1.5-T unit, in addition to CTA (CT angiography). Two independent readers evaluated each sequence independently in random order. Sensitivity, specificity, predictive values and inter-reader agreement were calculated for each sequence, excluding sequences judged inconclusive. Sensitivity was also calculated according to PE location. Contrast-enhanced angiographic sequences showed the highest sensitivity (82.9 and 89.7 %, reader 1 and reader 2, respectively), specificity (98.5 and 100 %) and agreement (kappa value 0.77). Unenhanced angiographic sequences, although less sensitive overall (68.7 and 76.4 %), were sensitive for the detection of proximal PE (92.7 and 100 %) and showed high specificity (96.1 and 99.1 %) and good agreement (kappa value 0.62). Perfusion sequences showed lower sensitivity (75.0 and 79.3 %), specificity (84.8 and 89.7 %) and agreement (kappa value 0.51), and a negative predictive value of 84.8 % at best. Compared with contrast-enhanced angiographic sequences, unenhanced sequences demonstrate lower sensitivity, except for proximal PE, but high specificity and agreement. The negative predictive value of perfusion sequences was insufficient to safely rule out PE. (orig.)

  4. Long-distance transport of 35S-sulphur in 3-year-old beech trees (Fagus sylvatica)

    35S-L-cysteine was fed to a mature leaf of 3-year-old beech trees via a flap. After 1 to 4 h the distribution of 35S-radioactivity was analysed in the leaves as well as the bark and wood of the trunk and the main root. Transport of 35S out of the fed leaf amounted to 0.3–1.2% of the total 35S taken up. The branches of the trees did not act as sink organs for the exported radioactivity. The main portion of the 35S-radioactivity transported out of the fed leaf was found in basipetal parts of the trunk. Only a small portion of 35S-radioactivity was transported in acropetal direction. The distribution of the 35S-radioactivity within the trunk showed a higher portion of 35S in the bark than in the wood. In both tissues, bark (70 to 80%) and wood (60 to 70%), the 35S was predominantly found in the HCl soluble fraction. However, 35S-cysteine, the compound fed to the leaves was not exported out of the fed leaf. Along the trunk 35S-cysteine was neither determined in bark nor in wood sections. The only low molecular mass S-compounds found was 35S-glutathione (GSH). The 35S-sulphate detected in bark and wood origined from cysteine oxidation in the leaf tissue and from contamination of the 35S-cysteine feeding solution. The ratio of GSH to sulphate decreased with increasing distance from the fed leaf. Apparently, 35S-radioactivity was transported as sulphate and GSH in the phloem in basipetal direction, but GSH was removed preferentially out of the phloem along the transport path. 35S-radioactivity exported out of the phloem and transported into the wood of the trunk was not retranslocated in the xylem. It may therefore be assumed that part of the 35S translocated was stored in ray cells, medullary sheath cells and/or pith parenchyma cells. Girdling experiments in which the bark of the trunk was peeled off basipetal to the branch containing the fed leaf support these assumptions. (author)

  5. Contrast-enhanced dynamic MR imaging of parasellar tumor using fast spin-echo sequence

    We have applied a new dynamic MRI technique that uses a fast spin-echo sequence to parasellar tumors. This sequence has less susceptible effect and better spatial resolution than a gradient echo sequence, providing faster images than a short spin-echo sequence does. Image was obtained in the coronal or sagittal plane using a 1.5T clinical MRI system, and then, dynamic MR images were acquired every 10 to 20 sec after administration of Gd-DTPA (0.1 mmol/kg). The subjects were 12 patients (5 microadenomas, 5 macroadenomas and 2 Rathke's cleft cysts) and 5 normal volunteers. As for volunteers, the cavernous sinus, pituitary stalk and posterior pituitary gland were contrasted on the first image, followed by visualization of the proximal portion adjacent to the junction of the infundibulum and the anterior pituitary gland, and finally by contrasting the distal portion of the anterior pituitary gland. There was a difference with respect to tumor contrast between microadenomas and macroadenomas. In the case of the macroadenomas, the tumor was contrasted at the same time as, or faster than the anterior pituitary gland, while with the microadenomas the tumor was enhanced later than the anterior pituitary gland. No enhancement with contrast medium was seen in Rathke's cleft cysts. In addition, it was possible to differentiate a recurrent tumor from a piece of muscle placed at surgery since the images obtained by the fast spin-echo sequence were clearer than those obtained by gradient echo sequence. (author)

  6. Sequence comparison for non-enhanced MRA of the lower extremity arteries at 7 Tesla.

    Sören Johst

    Full Text Available In this study three sequences for non-contrast-enhanced MRA of the lower extremity arteries at 7T were compared. Cardiac triggering was used with the aim to reduce signal variations in the arteries. Two fast single-shot 2D sequences, a modified Ultrafast Spoiled Gradient Echo (UGRE sequence and a variant of the Quiescent-Interval Single-Shot (QISS sequence were triggered via phonocardiogram and compared in volunteer examinations to a non-triggered 2D gradient echo (GRE sequence. For image acquisition, a 16-channel transmit/receive coil and a manually positionable AngioSURF table were used. To tackle B1 inhomogeneities at 7T, Time-Interleaved Acquisition of Modes (TIAMO was integrated in GRE and UGRE. To compare the three sequences quantitatively, a vessel-to-background ratio (VBR was measured in all volunteers and stations. In conclusion, cardiac triggering was able to suppress flow artifacts satisfactorily. The modified UGRE showed only moderate image artifacts. Averaged over all volunteers and stations, GRE reached a VBR of 4.18±0.05, UGRE 5.20±0.06, and QISS 2.72±0.03. Using cardiac triggering and TIAMO imaging technique was essential to perform non-enhanced MRA of the lower extremities vessels at 7T. The modified UGRE performed best, as observed artifacts were only moderate and the highest average VBR was reached.

  7. Synthesis of organic substances labelled with 14C and 35S

    After a brief history of the development of the Section des Molecules marquees of the French Atomic Energy Commission, the author gives an outline of the synthesis of the following labelled compounds: benzene 14C-6; phenyl-p-fluorophenyl, thienyl-2 β alanines β 14C; noradrenaline β 14C (arterenol β 14C), dotriacontane 14C-16-17, aminoethane sulfinic acid (hypotaurine 35S). (author)

  8. Response of a metastable superconducting grains suspension to irradiation by 35S decay electrons

    We report a measurement of the energy loss spectrum of electrons in the beta decay of 35S using a prototype detector composed of a suspension of 10-25 μm diameter tin grains at 2.3 K. Preliminary analyses suggests the capability of achieving sub-eV resolutions in such measurements, and that -contrary to conventional wisdoms - this is possible with both relatively large grains and a distribution of the grain sizes. (orig.)

  9. Distribution of [35S] taurine in mouse retina after intravitreal and intravascular injection

    The distribution of [35S] taurine in mouse retinae was studied by autoradiographic techniques after either intravitreal or intravascular injection. The route of injection did not affect the final localization. The major sites of label accumulation were the outer nuclear layer, the inner nuclear layer, and Mueller cell processes adjacent to the vitreal surface. The distribution was consistent with the interpretation that taurine was localized within two cellular compartments of mouse retina, photoreceptor cells and Mueller cells. (author)

  10. Biosynthesis of lutropin in ovine pituitary slices: incorporation of [35S]sulfate in carbohydrate units

    Sulfate incorporation into carbohydrate of lutropin (LH) has been studied in sheep pituitary slices using H2(35)SO4. Labeled ovine LH was purified to homogeneity by Sephadex G-100 and carboxymethyl-Sephadex chromatography from both the incubation medium and tissue extract. Autoradiography of the gel showed only two protein bands which comigrated with the alpha and beta subunits of ovine LH in both the purified ovine LH and the immunoprecipitate obtained with LH-specific rabbit antiserum. Furthermore, [35S]sulfate was also incorporated into several other proteins in addition to LH. The location of 35SO/sub 2-(4)/ in the oligosaccharides of ovine LH was evidenced by its presence in the glycopeptides obtained by exhaustive Pronase digestion. The location and the point of attachment of sulfate in the carbohydrate unit were established by the isolation of 4-O-[35S]sulfo-N-acetylhexosaminyl-glycerols and 4-O-[35S] sulfo-N-acetylglucosaminitol from the Smith degradation products and by the release of 35SO/sub 2-(4)/ by chondro-4-sulfatase. Thus, the present line of experimentation indicates the presence of sulfate on both the terminal N-acetylglucosamine and N-acetylgalactosamine in the oligosaccharide chains of the labeled ovine LH

  11. Cloning of a Nicotiana plumbaginifolia protoplast-specific enhancer-like sequence

    Horth, Marie; Negrutiu, Ioan; Burny, Arséne; Van Montagu, Marc; Herrera-Estrella, Luis

    1987-01-01

    We have isolated a 1.5-kb plant DNA fragment (called insert 7) from Nicotiana plumbaginifolia DNA that contains a protoplast-specific enhancer-like sequence. The presence of this sequence on a plasmid carrying a chimeric nos-npt-II gene conferring kanamycin resistance to plant cells, produces an overexpression of the npt-II gene during at least eight days after protoplast transformation. This effect on the expression of the nos promoter was independent of the orientation and was observed both...

  12. Preparation of titania nanotube-Cd0.65Zn0.35S nanocomposite by a hydrothermal sulfuration method for efficient visible-light-driven photocatalytic hydrogen production

    Li, Juan; Wu, Liangpeng; Long, Lizhen; Xi, Min; Li, Xinjun

    2014-12-01

    Titania nanotube-Cd0.65Zn0.35S nanocomposite (Cd0.65Zn0.35S-TiO2) was synthesized from titanate nanotubes for ion change of Cd2+ and Zn2+ followed by hydrothermal sulfuration treatment using thiourea as sulfur source. The Cd0.65Zn0.35S-TiO2 with enhanced crystallinity of TiO2 nanotube can be obtained by increasing hydrothermal temperature from 90 °C to 120 °C. And further increasing hydrothermal temperature to 150 °C, TiO2 nanotubes collapse and transform into irregular shaped particles. The photocatalytic activity for hydrogen production of the prepared Cd0.65Zn0.35S-TiO2 with different hydrothermal temperature was investigated under visible-light irradiation. The result shows that the Cd0.65Zn0.35S-TiO2 with hydrothermal temperature of 120 °C presents the highest hydrogen evolution rate and photostability, which can be attributed to a rapid charge transfer at the interface between Cd0.65Zn0.35S and TiO2 nanotube due to the increased crystallinity and unique 1-D nanotubular structure of TiO2.

  13. In vitro estimation of rumen protein degradability using 35S to label the bacterial mass

    An experiment was carried out in order to simplify a previously developed 15N-method for in vitro estimation of rumen protein degradability. Casein (Cas), whole soybeans (Sb) heated at 120oC for 20 min (SbTherm) and sunflower (Sfl) were incubated at 39oC for 4 hours in a water bathshaker with the following media: McDougall's buffer, strained and enriched with particle associated bacteria rumen fluid (2:1), rapidly (maltose, sucrose, glucose) and more slowly (pectin, soluble starch) degradable carbohydrates with final concentration of 815 mg/100 ml and 21.7 μCi/100 ml of35S (from Na235SO4). After the incubation had been ceased, a bacterial fraction was isolated through differential centrifugation and specific activity of bacterial (Bac) and high speed total solids (TS) nitrogen was measured. The ratio was used to calculate bacterial mass in TS and through the Kjeldahl nitrogen concentration in TS - the net bacterial growth (against control vessels without protein). The level of ammonia-N in the supernate after blank correction was used to find the ammonia-N released from protein degradation. The data showed that the rate (and extend) of degradation for the Cas (as a standard protein) was lower compared to those obtained through the 15N-method but it was higher than the rate derived through another in vitro method. The Cas equivalent of the Sb was higher than the figure we found in a previous experiment with solvent extracted soybean meal suggesting that the 35S-method underestimated the degradability of the Cas. After being tested on a wider range of foodstuffs, the proposed 35S-method might be considered as an alternative procedure which is less laborous than the 15N-method. (author)

  14. Detection of radioactive 35S at Fukushima and other Japanese sites

    Priyadarshi, Antra; Hill-Falkenthal, Jason; Thiemens, Mark H.; Yoshida, Naohiro; Toyoda, Sakae; Yamada, Keita; Mukotaka, Arata; Fujii, Ayako; Uematsu, Mitsuo; Hatakeyama, Shiro; Noguchi, Izumi; Nojiri, Yukihiro; Tanimoto, Hiroshi

    2013-01-01

    The Fukushima nuclear power plant was severely damaged by an earthquake and concomitant tsunami during March 2011. An effect of this disaster was secondary formation of radioactive 35S via the 35Cl(n,p)35S reaction, when neutrons from the partially melted reactor cores activated the coolant sea water. Here we report the first measurements of 35S in sulfate aerosols and rain water collected at six Japanese sampling sites, Hokkaido, Tsukuba, Kashiwa, Fuchu, Yokohama, and Fukushima, during March-September 2011. The measured 35SO42- concentrations in aerosols vary significantly. The Kashiwa (AORI) site shows the highest 35SO42- concentration (6.1 × 104 ± 200 atoms/m3) on 1 April 2011, which is nearly 100 times higher than the natural background activity. Considering the percentage loss of 35SO42- resulting from dry and wet deposition and dilution of the radiation plume in the boundary layer during transport, it was determined that the surface air concentration of 35SO42- at the Fukushima would have been 2.8 × 105 atoms/m3 during the week after the earthquake, which is in agreement with the model prediction [Priyadarshi et al.]. 35SO42- activity in rain water collected during March-May 2011 at Tokyo Tech Yokohama varies from 1.1 × 105 to 9.8 × 105 atoms/liter, whereas stream water collected near Fukushima was found to have 1.2 × 105 atoms/liter during April. Even after 6 months, 35SO42- activity remains very high (9.9 × 104 ± 770 atoms/m3) in the marine boundary layer in the Fukushima region, which implies that the reactor core was producing radioactive sulfur.

  15. Diffusion of sulfur 35S in β-Ni3S2

    The diffusion of 35S radioisotope in β-Ni3S2 polycrystals was studied at temperatures ranging from 848 to 893 K and at sulfur vapour pressure (5.6 x 10-6 - 3.2 x 10-4) Pa. The autoradiography and the tracer sectioning method were used. It was found that the bulk diffusion is the main process of sulfur transport. The activation energy of the diffusion equals (190±10) kJmol-1. Under the conditions used in the experiments the dominant anionic point defects are single-ionized vacancies and quasi-free electrons. (author)

  16. Formation of Triblock Copolymers via a Tandem Enhanced Spin Capturing-Nitroxide-Mediated Polymerization Reaction Sequence

    Junkers, Thomas; Zang, Lin; Wong, Edgar H. H.; Dingenouts, Nico; Barner-Kowollik, Christopher

    2011-01-01

    The preparation of ABA-type block copolymers via tandem enhanced spin capturing polymerization (ESCP) and nitroxide-mediated polymerization (NMP) processes is explored in-depth. Midchain alkoxyamine functional polystyrenes (M(n) = 6200, 12,500 and 19,900 g mol(-1)) were chain extended with styrene as well as tert-butyl acrylate at elevated temperature NMP conditions (T = 110 degrees C) generating a tandem ESCP-NMP sequence. Although the chain extensions and thus the block copolymer formation ...

  17. Enhanced method for flaws depth estimation in CFRP slabs from FDTC thermal contrast sequences

    Andrés David Restrepo Girón

    2015-01-01

    After the detection of internal defects in materials, the characterization of these plays a decisive role in order to establish the severity of these flaws. Finite difference thermal contrast (FDTC) is a new technique proposed recently for contrast enhancement in sequences of thermal images in order to allow the detection of internal flaws in composite slabs with greater probability of success. Besides FDTC, a criterion was also conceived for the estimation of the depth of the detected defect...

  18. MR of normal pancreas : comparison of five pulse sequences and enhancing patterns on dynamic imaging

    Jang, Hyun Jung; Kim, Tae Kyoung; Hong, Sung Hwan; Han, Joon Koo; Choi, Byung Ihn [Seoul National Univ. College of Medicine, Seoul (Korea, Republic of)

    1997-03-01

    To compare T1-weighted FLASH and turbo spin echo (SE) T2-weighted sequences with conventional T1- and T2-weighted sequences in imaging normal pancreas and to describe the enhancing patterns on dynamic MR imging. Forty-four patients with presumed hepatic hemangiomas were studied at 1.0T or 1.5T by using conventional SE sequences (T1-weighted, T2-weighted, and heavily T2-weighted), turbo-SE T2-weighted sequences, and breath-hold T1-weighted FLASH sequences acquired before, immediately on, and at 1, 2, 3, and 5 or 10 minutes after injection of a bolus of gadopentetate dimeglumine. No patients had either a history or its clinical features of pancreatic disease. Images were quantitatively analyzed for signal-difference-to noise ratios (SD/Ns) between the pancreas and peripancreatic fat. Percentage enhancement of the pancreas was measured on each dynamic MR image. Conspicuity of the pancreatic border was qualitatively evaluated according to a consensus, reached by three radiologists. Turbo-SE T2-weighted images had a significantly higher SD/N ratio (p<0.001) and better conspicuity of the pancreatic border (p<0.001) than SE T2- and heavily T2-weighted images;T1-weighted SE images had a significantly higher SD/N ratio than T1-weighted FLASH images (p<0.001), but there was no significant difference between tham in qualitative analysis (p=0.346). Percentage enhancement immediately on and at 1, 2, 3, 5, and 10 minutes after administration of contrast material was 39.9%, 44.5%, 42.9%, 40.8%, 36.3%, 29.9%, respectively, with peak enhancement at 1 minute. In MR imaging of normal pancreas, turbo-SE T2-weighted imaging is superior to SE T2- and heavily T2- weighted imaging, and SE T1-weighted imaging is superior to T1-weighted FLASH imaging. On serial gadolinium-enhanced FLASH imaging, normal pancreas shows peak enhancement at 1 minute.

  19. MR of normal pancreas : comparison of five pulse sequences and enhancing patterns on dynamic imaging

    To compare T1-weighted FLASH and turbo spin echo (SE) T2-weighted sequences with conventional T1- and T2-weighted sequences in imaging normal pancreas and to describe the enhancing patterns on dynamic MR imging. Forty-four patients with presumed hepatic hemangiomas were studied at 1.0T or 1.5T by using conventional SE sequences (T1-weighted, T2-weighted, and heavily T2-weighted), turbo-SE T2-weighted sequences, and breath-hold T1-weighted FLASH sequences acquired before, immediately on, and at 1, 2, 3, and 5 or 10 minutes after injection of a bolus of gadopentetate dimeglumine. No patients had either a history or its clinical features of pancreatic disease. Images were quantitatively analyzed for signal-difference-to noise ratios (SD/Ns) between the pancreas and peripancreatic fat. Percentage enhancement of the pancreas was measured on each dynamic MR image. Conspicuity of the pancreatic border was qualitatively evaluated according to a consensus, reached by three radiologists. Turbo-SE T2-weighted images had a significantly higher SD/N ratio (p<0.001) and better conspicuity of the pancreatic border (p<0.001) than SE T2- and heavily T2-weighted images;T1-weighted SE images had a significantly higher SD/N ratio than T1-weighted FLASH images (p<0.001), but there was no significant difference between tham in qualitative analysis (p=0.346). Percentage enhancement immediately on and at 1, 2, 3, 5, and 10 minutes after administration of contrast material was 39.9%, 44.5%, 42.9%, 40.8%, 36.3%, 29.9%, respectively, with peak enhancement at 1 minute. In MR imaging of normal pancreas, turbo-SE T2-weighted imaging is superior to SE T2- and heavily T2- weighted imaging, and SE T1-weighted imaging is superior to T1-weighted FLASH imaging. On serial gadolinium-enhanced FLASH imaging, normal pancreas shows peak enhancement at 1 minute

  20. Unexpected high 35S concentration revealing strong downward transport of stratospheric air during the monsoon transitional period in East Asia

    Lin, Mang; Zhang, Zhisheng; Su, Lin; Su, Binbin; Liu, Lanzhong; Tao, Jun; Fung, Jimmy C. H.; Thiemens, Mark H.

    2016-03-01

    October is the monsoon transitional period in East Asia (EA) involving a series of synoptic activities that may enhance the downward transport of stratospheric air to the planetary boundary layer (PBL). Here we use cosmogenic 35S in sulfate aerosols (35SO42-) as a tracer for air masses originating from the stratosphere and transported downward to quantify these mixing processes. From 1 year 35SO42- measurements (March 2014 to February 2015) at a background station in EA we find remarkably enhanced 35SO42- concentration (3150 atoms m-3) in October, the highest value ever reported for natural sulfate aerosols. A four-box 1-D model and meteorological analysis reveal that strong downward transport from the free troposphere is a vital process entraining aged stratospheric air masses to the PBL. The aged stratospheric masses are accumulated in the PBL, accelerating the SO2 transformation to SO42-. Implications for the tropospheric O3 budget and the CO2 biogeochemical cycle are discussed.

  1. [35S]autoradiographic study of sulfated GAG accumulation and turnover in embryonic mouse tooth germs

    The accumulation of sulfated glycosaminoglycans(GAG) in embryonic mouse molars before, during, and after terminal differentiation of odontoblasts was localized by [35S]autoradiography combined with the use of chondroitin ABC lyase. Much more sulfated GAG were accumulated in the dental papilla than in the dental epithelium. High incorporation of [35S]sulfate occurred at the epithelio-mesenchymal junction, which is the site of dental basement membrane and predentin. Before terminal differentiation of odontoblasts, the distribution of sulfated GAG was uniform at the basement membrane. After the onset of terminal differentiation of odontoblasts, much more sulfated GAG accumulated at the tip of principal cusps than at the apical (inferior) parts of cusps, and sulfated GAG were then found to be degraded more rapidly at the epithelio-mesenchymal junction than at other parts of the tooth germ. Thus regional variation in the rate of degradation of GAG exists in the tooth germs. Trypsin-isolated dental epithelia cultured in vitro synthesized a new basement membrane that could be labeled with [3H]glucosamine but not with 35SO4(-2). The epithelial-derived basal lamina contains little or no sulfatated GAG

  2. Preparation of titania nanotube-Cd{sub 0.65}Zn{sub 0.35}S nanocomposite by a hydrothermal sulfuration method for efficient visible-light-driven photocatalytic hydrogen production

    Li, Juan; Wu, Liangpeng; Long, Lizhen; Xi, Min; Li, Xinjun, E-mail: lixj@ms.giec.ac.cn

    2014-12-15

    Graphical abstract: - Highlights: • Cd{sub 0.65}Zn{sub 0.35}S-TiO{sub 2} nanotube is fabricated by a hydrothermal sulfuration method. • The hydrothermal temperature affects the composite's crystallinity and morphology. • Hydrothermal treated sample at 120 °C shows superior H{sub 2} evolution activity. • Enhanced activity is due to the increased crystallinity and 1-D tubular structure. - Abstract: Titania nanotube-Cd{sub 0.65}Zn{sub 0.35}S nanocomposite (Cd{sub 0.65}Zn{sub 0.35}S-TiO{sub 2}) was synthesized from titanate nanotubes for ion change of Cd{sup 2+} and Zn{sup 2+} followed by hydrothermal sulfuration treatment using thiourea as sulfur source. The Cd{sub 0.65}Zn{sub 0.35}S-TiO{sub 2} with enhanced crystallinity of TiO{sub 2} nanotube can be obtained by increasing hydrothermal temperature from 90 °C to 120 °C. And further increasing hydrothermal temperature to 150 °C, TiO{sub 2} nanotubes collapse and transform into irregular shaped particles. The photocatalytic activity for hydrogen production of the prepared Cd{sub 0.65}Zn{sub 0.35}S-TiO{sub 2} with different hydrothermal temperature was investigated under visible-light irradiation. The result shows that the Cd{sub 0.65}Zn{sub 0.35}S-TiO{sub 2} with hydrothermal temperature of 120 °C presents the highest hydrogen evolution rate and photostability, which can be attributed to a rapid charge transfer at the interface between Cd{sub 0.65}Zn{sub 0.35}S and TiO{sub 2} nanotube due to the increased crystallinity and unique 1-D nanotubular structure of TiO{sub 2}.

  3. Preparation of titania nanotube-Cd0.65Zn0.35S nanocomposite by a hydrothermal sulfuration method for efficient visible-light-driven photocatalytic hydrogen production

    Graphical abstract: - Highlights: • Cd0.65Zn0.35S-TiO2 nanotube is fabricated by a hydrothermal sulfuration method. • The hydrothermal temperature affects the composite's crystallinity and morphology. • Hydrothermal treated sample at 120 °C shows superior H2 evolution activity. • Enhanced activity is due to the increased crystallinity and 1-D tubular structure. - Abstract: Titania nanotube-Cd0.65Zn0.35S nanocomposite (Cd0.65Zn0.35S-TiO2) was synthesized from titanate nanotubes for ion change of Cd2+ and Zn2+ followed by hydrothermal sulfuration treatment using thiourea as sulfur source. The Cd0.65Zn0.35S-TiO2 with enhanced crystallinity of TiO2 nanotube can be obtained by increasing hydrothermal temperature from 90 °C to 120 °C. And further increasing hydrothermal temperature to 150 °C, TiO2 nanotubes collapse and transform into irregular shaped particles. The photocatalytic activity for hydrogen production of the prepared Cd0.65Zn0.35S-TiO2 with different hydrothermal temperature was investigated under visible-light irradiation. The result shows that the Cd0.65Zn0.35S-TiO2 with hydrothermal temperature of 120 °C presents the highest hydrogen evolution rate and photostability, which can be attributed to a rapid charge transfer at the interface between Cd0.65Zn0.35S and TiO2 nanotube due to the increased crystallinity and unique 1-D nanotubular structure of TiO2

  4. Genome dynamics of short oligonucleotides: the example of bacterial DNA uptake enhancing sequences.

    Mohammed Bakkali

    Full Text Available Among the many bacteria naturally competent for transformation by DNA uptake-a phenomenon with significant clinical and financial implications- Pasteurellaceae and Neisseriaceae species preferentially take up DNA containing specific short sequences. The genomic overrepresentation of these DNA uptake enhancing sequences (DUES causes preferential uptake of conspecific DNA, but the function(s behind this overrepresentation and its evolution are still a matter for discovery. Here I analyze DUES genome dynamics and evolution and test the validity of the results to other selectively constrained oligonucleotides. I use statistical methods and computer simulations to examine DUESs accumulation in Haemophilus influenzae and Neisseria gonorrhoeae genomes. I analyze DUESs sequence and nucleotide frequencies, as well as those of all their mismatched forms, and prove the dependence of DUESs genomic overrepresentation on their preferential uptake by quantifying and correlating both characteristics. I then argue that mutation, uptake bias, and weak selection against DUESs in less constrained parts of the genome combined are sufficient enough to cause DUESs accumulation in susceptible parts of the genome with no need for other DUES function. The distribution of overrepresentation values across sequences with different mismatch loads compared to the DUES suggests a gradual yet not linear molecular drive of DNA sequences depending on their similarity to the DUES. Other genomically overrepresented sequences, both pro- and eukaryotic, show similar distribution of frequencies suggesting that the molecular drive reported above applies to other frequent oligonucleotides. Rare oligonucleotides, however, seem to be gradually drawn to genomic underrepresentation, thus, suggesting a molecular drag. To my knowledge this work provides the first clear evidence of the gradual evolution of selectively constrained oligonucleotides, including repeated, palindromic and protein

  5. Studying of SIX5 protein expression responsible for Myotonic Dystrophy (DM1) using L- [α- 35S] Methionine

    Myotonic dystrophy type1 (DM1) is the most common inherited adult muscular disease. Clinical features of DM1 include myotonia, progressive muscle weakness and wasting, cataract, endocrine abnormalities, heart conduction defects, and reduced cognitive ability. DM1 is caused by the expansion of an unstable of CTG repeat in the 3'-UTR of the DMPK gene. The SIX5 gene is located downstream of DMPK. Expansions of the CTG repeat eliminate an enhancer element of SIX5.There may be several mechanisms by which the repeat expansion causes the DM1 phenotype and the SIX5 transcript has been shown in several studies to be reduced in DM1 patients in a repeat dependent manner. The aim of this work was to express SIX5 (A-B-C), SIX5 (A-C) and Drosophila Six4 proteins in E. Coli using L- [α- 35S] Methionine to detect the expressed protein. Achieving this aim would allow us to identify SIX5 target genes which could be responsible for the myotonic dystrophy clinical features. (author)

  6. Intronic and flanking sequences are required to silence enhancement of an embryonic beta-type globin gene.

    Wandersee, N J; Ferris, R C; Ginder, G D

    1996-01-01

    In the course of studying regulatory elements that affect avian embryonic rho-globin gene expression, the multipotential hematopoietic cell line K562 was transiently transfected with various rho-globin gene constructs containing or lacking an avian erythroid enhancer element. Enhanced levels of rho gene expression were seen from those constructs containing an enhancer element and minimal 5' or 3' flanking rho sequences but were not seen from enhancer-containing constructs that included extens...

  7. Use of Natural 35S to Trace Sulphate Cycling in Small Lakes, Flattops Wilderness Area, Colorado, U.S.A

    Measurements of the cosmogenically-produced 35S, a radioisotope of sulphur (t1/2 = 87 days), are reported for the Ned Wilson Lake watershed in Colorado. The watershed contains two small lakes and a flowing spring presumed to be representative of local ground water. The watershed is located in the Flattops Wilderness Area and the waters in the system have low alkalinity, making them sensitive to increases in acid and sulphate deposition. Time series of 35S measurements were made during the summers of 1995 and 1996 (July-September) at all three sites. The system is dominated by melting snow and an initial concentration of 16-20 mBq L-1 was estimated for snow melt based on a series of snow samples collected in the Rocky Mountains. The two lakes had large initial 35S concentrations in July, indicating that a large fraction of the lake water and sulphate was introduced by meltwater from that year's snowpack. In 1995 and 1996, 35S concentrations decreased more rapidly than could be accounted for by decay, indicating that other processes were affecting 35S concentrations. The most likely explanation is that exchange with sediments or the biota was removing 35S from the lake and replacing it with older sulphate devoid of 35S. In September of 1995 and 1996, 35S concentrations increased, suggesting that atmospheric deposition is important in the sulphate flux of these lakes in late summer. Sulphur-35 concentrations in the spring water were highly variable but never higher than 3.6 mBq L-1 and averaged 2 mBq L-1. Using a simple mixing model, it was estimated that 75% of the spring water was derived from precipitation of previous years

  8. Differentiation of Leptomeningeal and Vascular Enhancement on Post-contrast FLAIR MRI Sequence: Role in Early Detection of Infectious Meningitis

    Ahmad, Armeen; Azad, Sheenam; Azad, Rajiv

    2015-01-01

    Objective: To qualitatively and quantitatively differentiate leptomeningeal and vascular enhancement on Post-contrast Fluid Attenuated Inversion Recovery (PCFLAIR) sequence compared to post-contrast T1-weighted (PCT1W) sequence with fat suppression (FS) and evaluate its role in early detection of infectious meningitis.

  9. Antisense EGFR sequence enhances apoptosis in a human hepatoma cell line BEL—7404

    FUTAO; HELIU; 等

    1996-01-01

    Effects of antisense epidermal growth factor receptor (EGFR) sequence on apoptotic cell death were examined in a human hepatoma cell line BEL-7404 cells.In the cells of JX-1,a sub clone of BEL-7404 stably transfected with antisense EGFR vector (Cell Research,3:75,1993),an enhanced rate(9.5%) of spontaneous apoptosis was detected by flow cytometry,whereas the rates of spontaneous apoptosis in JX-0 cells,a sub-clone of BEL-7404 transfected by control vector,and the parent BEL-7404 transfected by control vector,and the parent BEL-7404 transfected by control vector,and the parent BEL-7404 cells were almost equal and about 1.7%.Serum-starvation for 72h increased the rate of apoptosis of JX-lcells up to 33.7%,while JX-0 and BEL-7404 cells,under the same condition,produced less than 5% of apoptotic cells.Observation with electron microscope demonstrated that condensation and fragmentation of chromatin and formation of apoptotic bodies often occurred in JX-1 cells,especially during serumstarvation.These results,combined with the data of DNA fragmentation Elisa test,suggested that antisense EGFR sequence enhances apoptosis in the human hepatoma cells.Comparison of intracellular Ca2+ level and the responsiveness of JX-1 cells to the induced action of EGF and tharpsigargin (TG) treatment with that of control JX-0 cells indicated that antisense egfr might interrupt the EGF/EGFR sigaling pathway resulting in the decreass of intracellular Ca2+ pool content as well as the responsiveness of these cells to the extracellular signals.These findings suggest that antisense EGFR either directly or indirectly regulates Ca2+ storage in endoplasmic reticulum,thereby enhances apoptosis in the human hepatoma cells.

  10. Localization of 3H-glucosamine and 35S-sulfate in the hamster retina

    The localization of 3H-glucosamine and 35S-sulfate in the hamster retina was studied by light and electron microscopic autoradiography. Exposed silver grains were concentrated in the nerve fiber, inner plexiform, outer plexiform and inner segment layers. The heaviest labeling was observed over the inner segment layer in the glucosamine experiment and over the nerve fiber layers in the sulfate experiment. Specimens from 12- and 16-day-old hamsters showed slightly heavier labeling than did those from adult animals. In the electron microscopic autoradiography using 3H-glucosamine, grains were initially associated with the retinal pigment epithelium, the inner segment and the Mueller cell and were subsequently displaced into the spaces between the retinal pigment epithelium and the outer segment and into the nerve fiber layers. The labelled sulfates in the electron microscopic autoradiography were seen in the nerve fiber, plexiform and photoreceptor layers. These data indicate that glycosaminoglycans may be synthesized in the retinal pigment epithelium, the inner segment and the Mueller cell. (author)

  11. Effect of anticonvulsant drugs on (/sup 35/S)t-butylbicyclophosphorothionate binding in vitro and ex vivo

    Pitkaenen, A.; Riekkinen, P.J.; Saano, V.; Tuomisto, L.

    1987-01-01

    Using several concentrations of eight anticonvulsant drugs in clinical use (carbamazepine, clonazepam, phenytoin, phenobarbital, ethosuximide, primidone, sodium valproate, and D,L-..gamma..-vinyl GABA), we studied their abilities in vitro to displace (/sup 35/S)t-butylbicyclophosphorothionate (/sup 35/S-TBPS) from its binding site in a homogenate of rat brain. Thereafter ethosuximide (150 mg/kg), phenobarbital (30 mg/kg), clonazepam (0.3 mg/kg), or phenytoin (100 mg/kg) was injected intraperitoneally into rats for 16-20 days; and the effect of drug administration on /sup 35/S-TBPS binding was studied in the cortex and hippocampus ex vivo. Phenobarbital (100 ..mu..M, P<0.001), ethosuximide (500 ..mu..M, P<0.001), and phenytoin (40 ..mu..M, P<0.001) decreased the specific /sup 35/S-TBPS binding in vitro by 10-16%. After drug administration of phenobarbital (concentration in plasma 168 ..mu..M), the number of binding sites decreased and the binding affinity (p<0.05) in the cortex increased. Other anticonvulsants did not modulate /sup 35/S-TBPS binding in vitro at the concentration analogous to therapeutic plasma levels or ex vivo at the dose used. These results suggest that the use of phenobarbital may modulate the TBPS binding site, but the role of the present findings in the anticonvulsant action of phenobarbital needs to be further studied.

  12. A plant 35S CaMV promoter induces long-term expression of luciferase in Atlantic salmon.

    Seternes, Tore; Tonheim, Tom C; Myhr, Anne I; Dalmo, Roy A

    2016-01-01

    The long-term persistence and activity of a naked plasmid DNA (pGL3-35S) containing a luc gene (reporter gene) controlled by a plant 35S CaMV promoter was studied in Atlantic salmon (Salmo salar L.) after injection. Atlantic salmon (mean weight 70 grams) were injected intramuscularly with 100 μg of plasmid DNA. Blood, different tissues and organs were sampled at different time points up to day 535 after injection. Southern blot analysis suggested the presence of extra-chromosomally open circular, linear and supercoiled topoforms of pGL3-35S at day 150 after injection. At day 536 open circular and supercoiled topoforms were detected. Luciferase activity was detected at the injection site up to 536 days post-injection of pGL3-35S, where it peaked at day 150 and decreased to approximately 17% of its maximum activity by day 536. Our study demonstrated that a plasmid containing the 35S promoter was able to induce expression of a reporter gene/protein in fish in vivo and that the plasmid DNA persisted for a prolonged time after intramuscular injection. PMID:27114167

  13. Fat-saturated, contrast-enhanced spin echo sequences in magnetic resonance tomographic diagnosis of peritoneal carcinosis

    Purpose: To evaluate contrast-enhanced, fat-saturated spin echo sequences for the detection of peritoneal carcinosis with MRI. Material and Methods: 61 patients, 35 with and 26 without peritoneal carcinosis, were examined with abdominal MRI. Fat-saturated, T1-weighted spin echo sequences were performed before and after administration of Gd-DTPA. In addition, 22 patients with peritoneal carcinosis were examined with contrast-enhanced abdominal CT. Results: 32 of 35 patients with peritoneal carcinosis demonstrated contrast enhancement of the visceral and 30 to 35 enhancement of the parietal peritoneum (91 and 86%, respectively). Wall thickening of the intestine or parietal peritoneum were noted in 21 and 20 of 35 patients (60 and 57%, respectively), ascites in 18 of 35 patients (51%). False positive contrast enhancement of the peritoneum was noted in 4 of 26 patients (15%). In the direct comparison of MRI and CT, 22 of 22 patients versus 7 of 22 patients showed contrast enhancement of the visceral peritoneum (100 and 32%, respectively). For other signs of peritoneal carcinosis (e.g., ascites, peritoneal seedings), no differences in diagnostic reliability were demonstrated. Conclusions: The use of fat-saturated, spin echo sequences facilitates the diagnosis of peritoneal carcinosis by artifact reduction and improved detection of peritoneal contrast enhancement. MRI with fat-saturated sequences was superior to CT. (orig.)

  14. In situ detection of a heat-shock regulatory element binding protein using a soluble short synthetic enhancer sequence

    Harel-Bellan, A.; Brini, A.T.; Farrar, W.L. (National Cancer Institute, Frederick, MD (USA)); Ferris, D.K. (Program Resources, Inc., Frederick, MD (USA)); Robin, P. (Institut Gustave Roussy, Villejuif (France))

    1989-06-12

    In various studies, enhancer binding proteins have been successfully absorbed out by competing sequences inserted into plasmids, resulting in the inhibition of the plasmid expression. Theoretically, such a result could be achieved using synthetic enhancer sequences not inserted into plasmids. In this study, a double stranded DNA sequence corresponding to the human heat shock regulatory element was chemically synthesized. By in vitro retardation assays, the synthetic sequence was shown to bind specifically a protein in extracts from the human T cell line Jurkat. When the synthetic enhancer was electroporated into Jurkat cells, not only the enhancer was shown to remain undegraded into the cells for up to 2 days, but also its was shown to bind intracellularly a protein. The binding was specific and was modulated upon heat shock. Furthermore, the binding protein was shown to be of the expected molecular weight by UV crosslinking. However, when the synthetic enhancer element was co-electroporated with an HSP 70-CAT reporter construct, the expression of the reporter plasmid was consistently enhanced in the presence of the exogenous synthetic enhancer.

  15. Repeated reunions and splits feature the highly dynamic evolution of 5S and 35S ribosomal RNA genes (rDNA in the Asteraceae family

    Garcia Sònia

    2010-08-01

    Full Text Available Abstract Background In flowering plants and animals the most common ribosomal RNA genes (rDNA organisation is that in which 35S (encoding 18S-5.8S-26S rRNA and 5S genes are physically separated occupying different chromosomal loci. However, recent observations established that both genes have been unified to a single 35S-5S unit in the genus Artemisia (Asteraceae, a genomic arrangement typical of primitive eukaryotes such as yeast, among others. Here we aim to reveal the origin, distribution and mechanisms leading to the linked organisation of rDNA in the Asteraceae by analysing unit structure (PCR, Southern blot, sequencing, gene copy number (quantitative PCR and chromosomal position (FISH of 5S and 35S rRNA genes in ~200 species representing the family diversity and other closely related groups. Results Dominant linked rDNA genotype was found within three large groups in subfamily Asteroideae: tribe Anthemideae (93% of the studied cases, tribe Gnaphalieae (100% and in the "Heliantheae alliance" (23%. The remaining five tribes of the Asteroideae displayed canonical non linked arrangement of rDNA, as did the other groups in the Asteraceae. Nevertheless, low copy linked genes were identified among several species that amplified unlinked units. The conserved position of functional 5S insertions downstream from the 26S gene suggests a unique, perhaps retrotransposon-mediated integration event at the base of subfamily Asteroideae. Further evolution likely involved divergence of 26S-5S intergenic spacers, amplification and homogenisation of units across the chromosomes and concomitant elimination of unlinked arrays. However, the opposite trend, from linked towards unlinked arrangement was also surmised in few species indicating possible reversibility of these processes. Conclusions Our results indicate that nearly 25% of Asteraceae species may have evolved unusual linked arrangement of rRNA genes. Thus, in plants, fundamental changes in intrinsic

  16. Recombinase Polymerase Amplification (RPA of CaMV-35S Promoter and nos Terminator for Rapid Detection of Genetically Modified Crops

    Chao Xu

    2014-10-01

    Full Text Available Recombinase polymerase amplification (RPA is a novel isothermal DNA amplification and detection technology that enables the amplification of DNA within 30 min at a constant temperature of 37–42 °C by simulating in vivo DNA recombination. In this study, based on the regulatory sequence of the cauliflower mosaic virus 35S (CaMV-35S promoter and the Agrobacterium tumefaciens nopaline synthase gene (nos terminator, which are widely incorporated in genetically modified (GM crops, we designed two sets of RPA primers and established a real-time RPA detection method for GM crop screening and detection. This method could reliably detect as few as 100 copies of the target molecule in a sample within 15–25 min. Furthermore, the real-time RPA detection method was successfully used to amplify and detect DNA from samples of four major GM crops (maize, rice, cotton, and soybean. With this novel amplification method, the test time was significantly shortened and the reaction process was simplified; thus, this method represents an effective approach to the rapid detection of GM crops.

  17. Recombinase polymerase amplification (RPA) of CaMV-35S promoter and nos terminator for rapid detection of genetically modified crops.

    Xu, Chao; Li, Liang; Jin, Wujun; Wan, Yusong

    2014-01-01

    Recombinase polymerase amplification (RPA) is a novel isothermal DNA amplification and detection technology that enables the amplification of DNA within 30 min at a constant temperature of 37-42 °C by simulating in vivo DNA recombination. In this study, based on the regulatory sequence of the cauliflower mosaic virus 35S (CaMV-35S) promoter and the Agrobacterium tumefaciens nopaline synthase gene (nos) terminator, which are widely incorporated in genetically modified (GM) crops, we designed two sets of RPA primers and established a real-time RPA detection method for GM crop screening and detection. This method could reliably detect as few as 100 copies of the target molecule in a sample within 15-25 min. Furthermore, the real-time RPA detection method was successfully used to amplify and detect DNA from samples of four major GM crops (maize, rice, cotton, and soybean). With this novel amplification method, the test time was significantly shortened and the reaction process was simplified; thus, this method represents an effective approach to the rapid detection of GM crops. PMID:25310647

  18. Resolving the impact of stratosphere-to-troposphere transport on the sulfur cycle and surface ozone over the Tibetan Plateau using a cosmogenic 35S tracer

    Lin, Mang; Zhang, Zhisheng; Su, Lin; Hill-Falkenthal, Jason; Priyadarshi, Antra; Zhang, Qianggong; Zhang, Guoshuai; Kang, Shichang; Chan, Chuen-Yu; Thiemens, Mark H.

    2016-01-01

    The Himalayas were recently identified as a global hot spot for deep stratosphere-to-troposphere transport (STT) in spring. Although the STT in this region may play a vital role in tropospheric chemistry, the hydrological cycle and aquatic ecosystems in Asia, there is no direct measurement of a chemical stratospheric tracer to verify and evaluate its possible impacts. Here we use cosmogenic 35S as a tracer for air masses originating in the stratosphere and transported downward. We measure concentrations of 35S in fresh surface snow and river runoff samples collected from Mount Everest in April 2013 to be more than 10 times higher than previously reported by any surface measurement, in support of the Himalayas as a gateway of springtime STT. In light of this result, measurements of 35SO2 and 35SO42- at Nam Co in spring 2011 are reanalyzed to investigate the magnitudes of stratospheric air masses from the Himalayas to the tropospheric sulfur cycle and surface O3 level over the Tibetan Plateau. A simple one-box model reveals that the oxidative lifetime of SO2 is reduced in aged STT plumes. Triple oxygen isotopic measurements of sulfate samples suggest that enhanced O3 levels may shift the oxidation pathway of SO2 in the troposphere, which may be constrained by further intensive sampling and measurements. Comparison with surface O3 measurements and traditional meteorological tracing methods shows that 35S is a potentially unique and sensitive tracer to quantify the contribution of stratospheric air to surface O3 levels in fresh or aged STT plumes.

  19. Video Enhancement and Dynamic Range Control of HDR Sequences for Automotive Applications

    Giovanni Ramponi

    2007-01-01

    Full Text Available CMOS video cameras with high dynamic range (HDR output are particularly suitable for driving assistance applications, where lighting conditions can strongly vary, going from direct sunlight to dark areas in tunnels. However, common visualization devices can only handle a low dynamic range, and thus a dynamic range reduction is needed. Many algorithms have been proposed in the literature to reduce the dynamic range of still pictures. Anyway, extending the available methods to video is not straightforward, due to the peculiar nature of video data. We propose an algorithm for both reducing the dynamic range of video sequences and enhancing its appearance, thus improving visual quality and reducing temporal artifacts. We also provide an optimized version of our algorithm for a viable hardware implementation on an FPGA. The feasibility of this implementation is demonstrated by means of a case study.

  20. A field study of the uptake of 35S and 14C into crops characteristic of the UK diet

    The uptake of 35S and 14C into crops characteristic of the UK diet was studied. Four common types of green vegetable, six common types of root vegetable and perennial ryegrass were grown in a garden plot in the environs of Hinkley Point Nuclear Power Station and the 35S and 14C contents of the crops were measured. Also measured were the corresponding air concentrations over the plot averaged over a range of time periods between sowing and harvesting. The results were analysed in terms of air to crop transfer factors for 35S and 14C and the implications of these for dose calculations were assessed for both collective dose and for a hypothetical critical group consuming a range of foods produced in situ. (author)

  1. Thermal effect on photolysis in 203Hg, 35S and 131I labeled mercury (II) iodide thiocyanate powders

    203Hg, 35S or 131I labeled mercury (II) iodide thiocyanate (HgISCN) powders were prepared, respectively. When the powders were exposed to sunlight, some parts of the crystals of the powders were decomposed and the decomposed atoms diffused toward crystal surface. This diffusion velocity was accelerated by thermal treatment of the darkened powders. The velocity is larger in order of 35S, 203Hg, 131I. Decomposed products consist of colloidal mercury, mercury iodide, mercury sulfide, sulfur dioxide and iodine. Mechanism of photochromism of HgISCN was discussed. (author)

  2. Control of gene conversion and somatic hypermutation by immunoglobulin promoter and enhancer sequences.

    Yang, Shu Yuan; Fugmann, Sebastian D; Schatz, David G

    2006-12-25

    It is thought that gene conversion (GCV) and somatic hypermutation (SHM) of immunoglobulin (Ig) genes occur in two steps: the generation of uracils in DNA by activation-induced cytidine deaminase, followed by their subsequent repair by various DNA repair pathways to generate sequence-diversified products. It is not known how either of the two steps is targeted specifically to Ig loci. Because of the tight link between transcription and SHM, we have investigated the role of endogenous Ig light chain (IgL) transcriptional control elements in GCV/SHM in the chicken B cell line DT40. Promoter substitution experiments led to identification of a strong RNA polymerase II promoter incapable of supporting efficient GCV/SHM. This surprising finding indicates that high levels of transcription are not sufficient for robust GCV/SHM in Ig loci. Deletion of the IgL enhancer in a context in which high-level transcription was not compromised showed that the enhancer is not necessary for GCV/SHM. Our results indicate that cis-acting elements are important for Ig gene diversification, and we propose that targeting specificity is achieved through the combined action of several Ig locus elements that include the promoter. PMID:17178919

  3. Bioaggregate of photo-fermentative bacteria for enhancing continuous hydrogen production in a sequencing batch photobioreactor

    Xie, Guo-Jun; Liu, Bing-Feng; Wang, Rui-Qing; Ding, Jie; Ren, Hong-Yu; Zhou, Xu; Ren, Nan-Qi

    2015-11-01

    Hydrogen recovery through solar-driven biomass conversion by photo-fermentative bacteria (PFB) has been regarded as a promising way for sustainable energy production. However, a considerable fraction of organic substrate was consumed for the growth of PFB as biocatalysts, furthermore, these PFB were continuously washed out from the photobioreactor in continuous operation because of their poor flocculation. In this work, PFB bioaggregate induced by L-cysteine was applied in a sequencing batch photobioreactor to enhance continuous hydrogen production and reduce biomass washout. The effects of the hydraulic retention time (HRT), influent concentration and light intensity on hydrogen production of the photobioreactor were investigated. The maximum hydrogen yield (3.35 mol H2/mol acetate) and production rate (1044 ml/l/d) were obtained at the HRT of 96 h, influent concentration of 3.84 g COD/l, and light intensity of 200 W/m2. With excellent settling ability, biomass accumulated in the photobioreactor and reached 2.15 g/l under the optimum conditions. Structural analysis of bioaggregate showed that bacterial cells were covered and tightly linked together by extracellular polymeric substances, and formed a stable structure. Therefore, PFB bioaggregate induced by L-cysteine is an efficient strategy to improve biomass retention capacity of the photobioreactor and enhance hydrogen recovery efficiency from organic wastes.

  4. Comparison of four enhancement strategies for aerobic granulation in sequencing batch reactors

    Aerobic granules were developed in four identical sequencing batch reactors (SBRs) with synthetic wastewater to compare different strategies for the enhancement of granulation. The SBRs were operated by (a) increasing organic loading rate in R1; (b) reducing settling time in R2; (c) extending starvation period in R3; and (d) increasing shear force in R4. The results showed that four operational strategies were able to enhance aerobic granulation successfully in SBR, but that also showed different effect on the granulation process and characteristics of mature aerobic granules. The rapidest granulation was observed by using short settling time (R2) and the granules had higher extracellular polymeric substance (EPS) than other reactors. Extended starvation period (R3) and high shear force (R4) resulted in longer granulation period and the granules with higher integrity and smaller size. Higher organic loading rate (R1) resulted in the granules with larger size and higher K value. The maximum specific COD removal rates (qmax) of the granules in all SBRs were at a similar level (0.13-0.16 g COD/h-g VSS) but the granules in R1 and R2 had higher apparent half rate constant (K) of 18 and 16 mg/L, than those in R3 and R4 (2.8 and 3.3 mg/L).

  5. GMAP and GSNAP for Genomic Sequence Alignment: Enhancements to Speed, Accuracy, and Functionality.

    Wu, Thomas D; Reeder, Jens; Lawrence, Michael; Becker, Gabe; Brauer, Matthew J

    2016-01-01

    The programs GMAP and GSNAP, for aligning RNA-Seq and DNA-Seq datasets to genomes, have evolved along with advances in biological methodology to handle longer reads, larger volumes of data, and new types of biological assays. The genomic representation has been improved to include linear genomes that can compare sequences using single-instruction multiple-data (SIMD) instructions, compressed genomic hash tables with fast access using SIMD instructions, handling of large genomes with more than four billion bp, and enhanced suffix arrays (ESAs) with novel data structures for fast access. Improvements to the algorithms have included a greedy match-and-extend algorithm using suffix arrays, segment chaining using genomic hash tables, diagonalization using segmental hash tables, and nucleotide-level dynamic programming procedures that use SIMD instructions and eliminate the need for F-loop calculations. Enhancements to the functionality of the programs include standardization of indel positions, handling of ambiguous splicing, clipping and merging of overlapping paired-end reads, and alignments to circular chromosomes and alternate scaffolds. The programs have been adapted for use in pipelines by integrating their usage into R/Bioconductor packages such as gmapR and HTSeqGenie, and these pipelines have facilitated the discovery of numerous biological phenomena. PMID:27008021

  6. Effect of various treatments of protein on rumen metabolism and incorporation of 35S in the microbial protein

    A replicated 4x4 Latin square design involving switch over experiment was conducted on four adult bucks fitted with rumen fistula. The animals were offered concentrate mixture with 15 percent DCP and 67 percent TDN containing untreated and treated protein in control and experimental rations, respectively. There was a significant difference (P 35S. (author)

  7. Buffer solutes as stabilizers of 35S-amino acids: A study of volatility, radiochemical purity and biological activity

    It is well established that volatile radioactive compounds are associated with preparations of 35S-amino acids, but misconceptions still exist. The addition of stabilizers to these preparations has created a false sense of security regarding the evolution of these volatiles from cell culture medium. Experiments presented here show that the commercially used buffers L-lysine, tricine and 3,4-pyridine-dicarboxylic acid all reduced, but did not eliminate, the evolution of 35S volatiles from tissue culture medium. In each case, the rate of release was approximately 12 nCi/mCi/day (or 0.001%), compared to approximately 44 nCi/mCi/day in the buffer's absence. None of six alternative buffers tested are more effective than the commercially used buffers. We found that the release of labeled volatiles is dependent on the nature of the culture medium, but is no greater from crude 35S-amino acid preparations than from purified 35S-methionine. These commercially used buffers are also effective in maintaining radiochemical purity and are compatible with biological activity; however, because these buffers do not entirely eliminate the release of radioactivity into the gas phase, routine safe-handling procedures should be practiced when working with these products

  8. Metabolism and distribution of 14C- and 35S-labeled carbon disulfide in immature rats of different ages

    The metabolism and distribution of 14C- and 35S-CS2 was examined in 1-, 5-, 10-, 20-, 30-, and 40-day-old rats. During a 3-hr period following an ip dose of 14C-CS2, 58-83% of the dose was expired as CS2 and 4-9% was metabolized to expired CO2 depending on age. Thirty- and forty-day-old rats metabolized significantly more CS2 to CO2 and expired significantly less CS2 than 1- through 20-day-old rats. At the end of the measured expiration period, only biotransformation products of CS2, which were in part covalently bound, remained in tissues from rats of all ages. Tissue levels of 35S-CS2-derived radioactivity exceeded levels of 14C-CS2-derived radioactivity indicating that sulfur metabolites free from the carbon atom of CS2 were formed in rats as young as 1 day of age. The 35S-CS2-derived radioactivity per g of tissue and thus 35S covalently bound to tissue protein was significantly higher in 1- through 20-day-old rats than in 30- and 40-day-old rats. Twenty-four hr after dosing, up to 13 times more 35S-labeled metabolites were covalently bound in organs from 1-day-old rats than in similar organs from 40-day-old rats. The results showed that elimination of the biotransformation products of CS2, in particular the covalently binding sulfur metabolites, was prolonged in newborn rats in comparison to 40-day-old rats

  9. Metabolism of 35S- and 14C-labeled 1-methyl-2-mercaptoimidazole in vitro and in vivo

    We previously described an in vitro incubation system for studying the mechanism of inhibition of thyroid peroxidase (TPO)-catalyzed iodination by the antithyroid drug 1-methyl-2-mercaptoimidazole (MMI). Inhibition of iodination in this system may be reversible or irreversible, depending on the relative concentrations of iodide and MMI and on the TPO concentration. Metabolism of the drug occurs under both conditions, and in the present investigation we used 35S- and 14C-labeled MMI together with reverse phase HPLC to examine the metabolic products associated with reversible and irreversible inhibition of iodination by MMI. Under conditions of reversible inhibition, MMI was rapidly metabolized and disappeared completely from the incubation mixture. With [35S]MMI, the earliest detectable 35S-labeled product was MMI disulfide, which reached a peak after a few minutes and then declined to undetectable levels. Coincident with the decrease in disulfide was the appearance of two 35S peaks, the major one corresponding to sulfate/sulfite, and the other to a component eluting at 7.5 min. Similar results were obtained for the disulfide and for the 7.5 min metabolite with [14C]MMI. The major 14C-labeled metabolite containing no S appeared to be 1-methylimidazole. Under conditions of irreversible inhibition, MMI disulfide was also the earliest detectable 35S-labeled metabolite. However, MMI decreased more slowly, and after reaching a nadir at about 6 min returned gradually to a level about halfway between the initial and the minimum value. The reformation of MMI appeared to involve the nonenzymatic disproportionation of MMI disulfide. Formation of the 7.5 min peak was also observed, but there was no formation of sulfate/sulfite

  10. Effect of BRCA2 sequence variants predicted to disrupt exonic splice enhancers on BRCA2 transcripts

    Brewster Brooke L

    2010-05-01

    Full Text Available Abstract Background Genetic screening of breast cancer patients and their families have identified a number of variants of unknown clinical significance in the breast cancer susceptibility genes, BRCA1 and BRCA2. Evaluation of such unclassified variants may be assisted by web-based bioinformatic prediction tools, although accurate prediction of aberrant splicing by unclassified variants affecting exonic splice enhancers (ESEs remains a challenge. Methods This study used a combination of RT-PCR analysis and splicing reporter minigene assays to assess five unclassified variants in the BRCA2 gene that we had previously predicted to disrupt an ESE using bioinformatic approaches. Results Analysis of BRCA2 c.8308 G > A (p.Ala2770Thr by mRNA analysis, and BRCA2 c.8962A > G (p.Ser2988Gly, BRCA2 c.8972G > A (p.Arg2991His, BRCA2 c.9172A > G (p.Ser3058Gly, and BRCA2 c.9213G > T (p.Glu3071Asp by a minigene assay, revealed no evidence for aberrant splicing. Conclusions These results illustrate the need for improved methods for predicting functional ESEs and the potential consequences of sequence variants contained therein.

  11. An imaging protocol for dynamic contrast-enhanced breast MRI with 3.0T, using sagittal sequence interleaved between axial sequences

    Park, Ji Eun; Lee, Jee Eun; Cha, Eun Suk [Dept. of Radiology, Ewha Womans Un:iversity School of Medicine, Seoul (Korea, Republic of)], e-mail: escha@ewha.ac.kr; Hwang, Ji-Young [Dept. of Radiology, Kangnam Sacred Hospital, Hallym Univ., Seoul (Korea, Republic of)

    2013-07-15

    Background: B1 transmission-field inhomogeneity has been reported at 3.0 Tesla (T) breast imaging. Enhancement measurements of breast cancers at 3.0T may be insufficient for some patients and improvements in imaging protocols are needed. Purpose: To quantify B1 inhomogeneities in normal tissue and malignant masses at 3.0T breast MR imaging and to evaluate effect of an imaging protocol using an interleaved sagittal sequence in dynamic contrast-enhanced MRI (DCE-MRI). Material and Methods: A total of 76 patients were included who underwent DCE-MRI of the breast at 3.0T with an imaging protocol consisting of 1st, 2nd, and 4-6th bilateral axial sequences, and 3rd and 7th unilateral sagittal sequences. Signal intensity (SI) of normal breast tissue was measured at nipple level in four bilateral locations (anterior, posterior, medial, and lateral). Mean whole breast and location specific SI were calculated and compared between right and left breast using a paired t-test. All malignant masses were classified into three groups according to tumor size on MRI ({<=}2 cm, 2-4 cm, >4 cm). SI of malignant masses was measured independently on axial and sagittal sequences. The axial-sagittal SI gap in each mass was calculated and difference between right and left breast was compared using the t test. Size of each malignant mass was compared with pathologic findings to assess performance of the imaging protocol. Results: SI of normal breast tissue were lower for the right breast (R-L difference, -91.9; P < 0.0001) and in all four locations (anterior, P < 0.01; posterior, P < 0.01; medial, P < 0.0001; lateral, P < 0.0001). SI of malignant masses were lower for the right breast among same size of the lesions (P < 0.0001), particularly < 4 cm (P < 0.0001). Decreased right to left difference in SI was produced with an interleaved sagittal sequence, as axial-sagittal gap of malignant masses was significant when tumor locates on the right side (P < 0.001). The concordance rate in

  12. Chunk concatenation evolves with practice and sleep-related enhancement consolidation in a complex arm movement sequence

    Blischke Klaus

    2016-06-01

    Full Text Available This paper addresses the notion of chunk concatenation being associated with sleep-related enhancement consolidation of motor sequence memory, thereby essentially contributing to improvements in sequence execution speed. To this end, element movement times of a multi-joint arm movement sequence incorporated in a recent study by Malangré et al. (2014 were reanalyzed. As sequence elements differed with respect to movement distance, element movement times had to be purged from differences solely due to varying trajectory lengths. This was done by dividing each element movement time per subject and trial block by the respective “reference movement time” collected from subjects who had extensively practiced each sequence element in isolation. Any differences in these “relative element movement times” were supposed to reflect element-specific “production costs” imposed solely by the sequence context. Across all subjects non-idiosyncratic, lasting sequence segmentation was shown, and four possible concatenation points (i.e. transition points between successive chunks within the original arm movement sequence were identified. Based on theoretical suppositions derived from previous work with the discrete sequence production task and the dual processor model (Abrahamse et al., 2013, significantly larger improvements in transition speed occurring at these four concatenation points as compared to the five fastest transition positions within the sequence (associated with mere element execution were assumed to indicate increased chunk concatenation. As a result, chunk concatenation was shown to proceed during acquisition with physical practice, and, most importantly, to significantly progress some more during retention following a night of sleep, but not during a waking interval.

  13. Detection of viral genomes in the liver by in situ hybridization using 35S-, bromodeoxyuridine-, and biotin-labeled probes

    Methods employing 35S-, biotin-, and bromodeoxyuridine (BrdUrd)-labeled DNA probes were compared for the detection of hepatitis B virus (HBV) and cytomegalovirus (CMV) in the liver. The results demonstrate that: 1) HBV can be detected reliably only by the use of radiolabeled probes, whereas methods employing nonradioactive probes obviously are not sensitive enough for this virus. The use of 35S-labeled probes shortens the exposure times considerably in comparison to tritiated probes. 2) Biotin-labeled probes are of limited value for in situ hybridization on liver tissues because the presence of endogenous avidin-binding activity often leads to false positive results. 3) Brd-Urd-labeled probes are a useful alternative to biotinylated probes for the detection of CMV. In comparison with biotinylated probes, BrdUrd-labeled probes produce a specific signal of similar staining intensity in the absence of background staining in the liver

  14. Effect of salseed-meal tannins on protein synthesis, 35S incorporation and cellulose digestibility by rumen microbes in vitro

    Tannins from seed-meal of sal (Shorea robusta Gaertn. F.) were fractionated after treatments into ethyl acetate (EA) and lead acetate (LA) fractions. In trial 1, incorporation of 35S from (NH4)235SO4 into microbial protein declined due to the effect of both 2% EA and LA fractions as compared to control. Microbial protein synthesis was depressed significantly (P 35S incorporation and cellulolysis at all levels of both the tannin fractions. It may be inferred that both the tannin fractions from salseed-meal showed antimicrobial activity and LA fraction seems to be more deleterious than EA fraction for rumen metabolism. The experiments were conducted in vitro using rumen liquor of a non-pregnant dry cow having permanent rumen fistula. (auth.)

  15. Neem seed meal utilization for growth and its influence on 35S in incorporation into microbial protein

    Twelve buffalo-calves in two groups (1 and 2) were fed a conventional concentrate diet and a diet containing 20 percent neem seed meal, respectively. The buffalo calves of group 1 gained 15.5 kg more in a period of 119 days than the group 2 with corresponding DM consumptions of 3.35 kg per day. In another experiment, two in vitro trials were conducted in duplicate with neem seed meal at the levels of 5.9, 11.1 and 20.0 percent along with required substrates and 35S utilization was estimated in microbial protein using Na235SO4. Apparently there were no differences in 35S incorporation between different treatments indicating thereby that nimbin and its derivatives might not have any adverse effect on microbial protein synthesis. (author)

  16. Real-time polymerase chain reaction detection of cauliflower mosaic virus to complement the 35S screening assay for genetically modified organisms.

    Cankar, Katarina; Ravnikar, Maja; Zel, Jana; Gruden, Kristina; Toplak, Natasa

    2005-01-01

    Labeling of genetically modified organisms (GMOs) is now in place in many countries, including the European Union, in order to guarantee the consumer's choice between GM and non-GM products. Screening of samples is performed by polymerase chain reaction (PCR) amplification of regulatory sequences frequently introduced into genetically modified plants. Primers for the 35S promoter from Cauliflower mosaic virus (CaMV) are those most frequently used. In virus-infected plants or in samples contaminated with plant material carrying the virus, false-positive results can consequently occur. A system for real-time PCR using a TaqMan minor groove binder probe was designed that allows recognition of virus coat protein in the sample, thus allowing differentiation between transgenic and virus-infected samples. We measured the efficiency of PCR amplification, limits of detection and quantification, range of linearity, and repeatability of the assay in order to assess the applicability of the assay for routine analysis. The specificity of the detection system was tested on various virus isolates and plant species. All 8 CaMV isolates were successfully amplified using the designed system. No cross-reactivity was detected with DNA from 3 isolates of the closely related Carnation etched ring virus. Primers do not amplify plant DNA from available genetically modified maize and soybean lines or from different species of Brassicaceae or Solanaceae that are natural hosts for CaMV. We evaluated the assay for different food matrixes by spiking CaMV DNA into DNA from food samples and have successfully amplified CaMV from all samples. The assay was tested on rapeseed samples from routine GMO testing that were positive in the 35S screening assay, and the presence of the virus was confirmed. PMID:16001857

  17. Method for the typing of Clostridium difficile based on polyacrylamide gel electrophoresis of (/sup 35/S)methionine-labeled proteins

    Tabaqchali, S.; O' Farrell, S.; Holland, D.; Silman, R.

    1986-01-01

    A typing method for Clostridium difficile based on the incorporation of (/sup 35/S)methionine into cellular proteins, their separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and their visualization by autoradiography is described. On analysis of the radiolabeled-protein profiles, nine distinct groups were observed (A to E and W to Z). The method, which is simple, reproducible, and readily expandable, has been applied in epidemiological studies to demonstrate cross-infection and hospital acquisition of C. difficile.

  18. Method for the typing of Clostridium difficile based on polyacrylamide gel electrophoresis of [35S]methionine-labeled proteins.

    Tabaqchali, S.; O'Farrell, S; Holland, D.; Silman, R

    1986-01-01

    A typing method for Clostridium difficile based on the incorporation of [35S]methionine into cellular proteins, their separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and their visualization by autoradiography is described. On analysis of the radiolabeled-protein profiles, nine distinct groups were observed (A to E and W to Z). The method, which is simple, reproducible, and readily expandable, has been applied in epidemiological studies to demonstrate cross-infection a...

  19. Method for the typing of Clostridium difficile based on polyacrylamide gel electrophoresis of [35S]methionine-labeled proteins

    A typing method for Clostridium difficile based on the incorporation of [35S]methionine into cellular proteins, their separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and their visualization by autoradiography is described. On analysis of the radiolabeled-protein profiles, nine distinct groups were observed (A to E and W to Z). The method, which is simple, reproducible, and readily expandable, has been applied in epidemiological studies to demonstrate cross-infection and hospital acquisition of C. difficile

  20. The distribution of 35S-sulfadiazine and 14C-trimethoprim in rainbow trout, Salmo gairdneri

    The distribution of 35S-labelled sulfadiazine and 14C-labelled trimethoprim was studied in rainbow trout by use of whole body autoradiography and liquid scintillation. As compared to mammals, gastrointestinal absorption and elimination were slow. Accumulation in the skin and the uveal tract of the eye was observed for both drugs tested. The results also indicated that the bile was an important route of excretion. Considerable radioactivity was still present in the skin at 144 hr. survival time. (author)

  1. Studies on uptake and translocation of sulphur in oil seed rape plant using 35S tracer technique

    The experiments were conducted to investigate the uptake and translocation of 35S in oil seed rape at different stages. The results showed that uptake of sulphur from soil by rape plant were 23.59%, 31.65% and 49.13% of sulphur applied at transplanting, bolting and flowering stage, respectively. About 40% of 35S taken from soil was located in seeds for transplanting and flowering stage applying. About 20% of sulphur applied on top four leaves at various stages of growth was remained in labelled leaves and 21%∼23% was transported to pods with majority in branch pods. 40%∼60% of sulphur which was applied on surface of pods at lower stem at different time after flowering was remained in shell of labelled pods and 13%∼16.8% of that was transferred into seeds of labelled pods. Recoveries of labelled sulphur applied on pods from seeds of pods at branches were less than 10%. The transportation of 35S from labelled sites to seeds of labelled pods and unlabelled pods was declined sharply as labelling was carried out at 6 weeks after flowering

  2. Study on kinetic degradation in soil and horizontal transfer of bt gene by 35S isotopic tracing method

    In this study, 35S isotopic tracing method was applied to investigate kinetic degradation of bt gene from Bt transgenic rice TT51 in two different soil and possibility of its horizontal transfer into soil bacteria as well. Results showed that, during 30 d of aerobic incubation, it was indicated that 35S-Bt gene was not horizontally transferred into soil microorganisms. The aerobic soil degradation dynamics significantly followed a first-order dissipation pattern for bt gene. After 30 d of incubation, the amount of bt gene reached 9.32% of applied radioactivity for the fluvio-marine yellow loamy soil and 9.92% for the fluvio-aquatic soil, respectively. The half-lives in two soils were 3.53 d for the former soil and 5. 77 d for the latter soil, which means that bt gene was more easily degradable in the weak acidic soil. The use of 35S labeling proved to be valuable; it served the purpose of validating the rigorousness of experimental protocols, and provided insights into the soil environmental safety assessment for Bt transgenic rice. (authors)

  3. An enhanced method for sequence walking and paralog mining: TOPO® Vector-Ligation PCR

    Davis Thomas M

    2010-03-01

    Full Text Available Abstract Background Although technological advances allow for the economical acquisition of whole genome sequences, many organisms' genomes remain unsequenced, and fully sequenced genomes may contain gaps. Researchers reliant upon partial genomic or heterologous sequence information require methods for obtaining unknown sequences from loci of interest. Various PCR based techniques are available for sequence walking - i.e., the acquisition of unknown DNA sequence adjacent to known sequence. Many such methods require rigid, elaborate protocols and/or impose narrowly confined options in the choice of restriction enzymes for necessary genomic digests. We describe a new method, TOPO® Vector-Ligation PCR (or TVL-PCR that innovatively integrates available tools and familiar concepts to offer advantages as a means of both targeted sequence walking and paralog mining. Findings TVL-PCR exploits the ligation efficiency of the pCR®4-TOPO® (Invitrogen, Carlsbad, California vector system to capture fragments of unknown sequence by creating chimeric molecules containing defined priming sites at both ends. Initially, restriction enzyme-digested genomic DNA is end-repaired to create 3' adenosine overhangs and is then ligated to pCR4-TOPO vectors. The ligation product pool is used directly as a template for nested PCR, using specific primers to target orthologous sequences, or degenerate primers to enable capture of paralogous gene family members. We demonstrated the efficacy of this method by capturing entire coding and partial promoter sequences of several strawberry Superman-like genes. Conclusions TVL-PCR is a convenient and efficient method for DNA sequence walking and paralog mining that is applicable to any organism for which relevant DNA sequence is available as a basis for primer design.

  4. Differential labeling of the subunits of respiratory complex III with [3H]succinic anhydride, [14C]succinic anhydride, and p-diazobenzene-[35S]sulfonate

    Exposure of antimycin-treated Complex III (ubiquinol-cytochrome c reductase) purified from bovine heart mitochondria to [3H]succinic anhydride plus [35S]p-diazobenzenesulfonate (DABS) resulted in somewhat uniform relative labeling of the eight measured subunits of the complex by [3H]succinic anhydride. In contrast, relative labeling by [35S]DABS was similar to [3H]succinic anhydride for the subunits of high molecular mass, i.e., core proteins, cytochromes, and the iron-sulfur protein, but greatly reduced for the polypeptides of molecular mass below 15 kDa. With Complex II depleted in the iron-sulfur protein the relative labeling of core protein I by exposure of the complex to [3H]succinic anhydride was significantly enhanced, whereas labeling of the polypeptides represented by SDS-PAGE bands 7 and 8 was significantly inhibited. Dual labeling of the subunits of Complex III by 14C- and 3H-labeled succinic anhydride before and after dissociation of the complex by sodium dodecyl sulfate, respectively, was measured with the complex in its oxidized, reduced, and antimycin-inhibited states. Subunits observed to be most accessible or reactive to succinic anhydride were core protein II, the iron-sulfur protein, and polypeptides of SDS-PAGE bands 7,8, and 9. Two additional polypeptides of molecular masses 23 and 12kDa, not normally resolved by gel-electrophoresis, were detected. Reduction of the complex resulted in a significant change of 14C/3H labeling ratio of core protein only, whereas treatment of the complex with antimycin resulted in decreases in 14C/3H labeling ratios of core proteins I and II, cytochrome c1, and a polypeptide of molecular mass 13kDa identified as an antimycin-binding protein

  5. The Effects of a High-Probability Request Sequencing Technique in Enhancing Transition Behaviors

    Banda, Devender R.; Kubina, Richard M., Jr.

    2006-01-01

    In this study, an autism support teacher used a high-probability request sequencing technique to help a middle-school student with autism engage in three transition behaviors. High probability request sequencing refers to a procedure in which 2 to 3 preferred questions, highly associated with compliance, are rapidly given before presenting a low…

  6. SPIO-enhanced MR imaging for HCC detection in cirrhotic patient : comparison of various techniques for optimal sequence selection

    To compare the efficacy of breathhold and non-breathhold sequences in the detection of hepatocellular carcinoma (HCC) in cirrhotic patients using superparamagnetic iron oxide (SPIO)-enhanced MR imaging, and to determine the optimal sequence combination. By means of unenhanced and iron-oxide-enhanced MRI, 29 patients with 49 nodular HCCs were evaluated for the presence of HCC nodules. Twenty-one were male and eight were female, and their ages ranged from 38 to 71 (mean, 56) years. Eight different MR sequences were used, including four non-breath-hold sequences and four breath-hold, and images were obtained before and after the administration of SPIO particles. Non-breath-hold sequences included T2-, proton density-weighted SE, and TSE imaging, while breath-hold sequences comprised T1-weighted fast low-angle shot (T1w FLASH), half-Fourier acquisition single shot turbo spine echo (HASTE), T2-weighted fast imaging with steady-state free precession (T2*wFISP) and T2-weighted breath-hold TSE (T2wBHTSE). Image analysis involved both quantitative and qualitative analysis. The quantitative parameters calculated were signal-to noise (S/N) ratios for livers and tumors, contrast to noise (C/N) ratios for tumors seen on precontrast and postcontrast images, and percentage of signal intensity loss (PSIL) after SPIO injection. Images were analysed qualitatively in terms of image artifacts and lesion conspicuity, and prior to calculating sensitivity, the number of lesions detected using various pulse sequences were counted. SPIO had a marked effect on liver S/N ratio but a minimal effect on tumor S/N ratio. PSIL was best in T2*wFISP images, while T2wSE images showed the second-best results (p less than 0.05). Tumor-to-liver C/N values were also highest with T2*wFISP, while T2wTSE and HASTE images were next. Qualitative study showed that non-breath hold images and FISP were better than breath hold images in terms of lesion conspicuity. The latter, however, were much better than non

  7. Detection of the 35S promoter in transgenic maize via various isothermal amplification techniques: a practical approach.

    Zahradnik, Celine; Kolm, Claudia; Martzy, Roland; Mach, Robert L; Krska, Rudolf; Farnleitner, Andreas H; Brunner, Kurt

    2014-11-01

    In 2003 the European Commission introduced a 0.9% threshold for food and feed products containing genetically modified organism (GMO)-derived components. For commodities containing GMO contents higher than this threshold, labelling is mandatory. To provide a DNA-based rapid and simple detection method suitable for high-throughput screening of GMOs, several isothermal amplification approaches for the 35S promoter were tested: strand displacement amplification, nicking-enzyme amplification reaction, rolling circle amplification, loop-mediated isothermal amplification (LAMP) and helicase-dependent amplification (HDA). The assays developed were tested for specificity in order to distinguish between samples containing genetically modified (GM) maize and non-GM maize. For those assays capable of this discrimination, tests were performed to determine the lower limit of detection. A false-negative rate was determined to rule out whether GMO-positive samples were incorrectly classified as GMO-negative. A robustness test was performed to show reliable detection independent from the instrument used for amplification. The analysis of three GM maize lines showed that only LAMP and HDA were able to differentiate between the GMOs MON810, NK603, and Bt11 and non-GM maize. Furthermore, with the HDA assay it was possible to realize a detection limit as low as 0.5%. A false-negative rate of only 5% for 1% GM maize for all three maize lines shows that HDA has the potential to be used as an alternative strategy for the detection of transgenic maize. All results obtained with the LAMP and HDA assays were compared with the results obtained with a previously reported real-time PCR assay for the 35S promoter in transgenic maize. This study presents two new screening assays for detection of the 35S promoter in transgenic maize by applying the isothermal amplification approaches HDA and LAMP. PMID:24880871

  8. Comparison of the activity of salt and 35S promoters in transgenic calli of sorghum (Sorghum bicolor L. (Moench))

    Use of specific promoters under clearly defined conditions in transgenic plants is valuable for determining gene expression. In this study, we used the Salt promoter derived from the rice variety Taichung native-1 and linked to the uidA gene in plasmid pGVB310, and compared its activity with that of the 35S promoter in the transformed calli of sorghum. The 35S promoter, as the constitutive promoter, is also linked to the uidA gene in plasmid p35SGus. Plasmid pGVB310 was transferred to sorghum cell suspension using biolistic bombardment in the same way as co-transformation was carried out for plasmid p35SGus with pROB5 containing the hpt gene as the selectable marker. A transformed callus was selected using 50 mg/L of hygromycin in callus medium for 3 weeks. The resistant colonies obtained during selection were then transferred to a fresh callus medium containint 50 mg/L of hygromycin. A histochemical Gus assay was performed on those calli that were resistant to 50 mg/L of hygromycin; the results showed that a blue colour appeared in the transformed calli after 5 hours of incubation at 37 deg C. A fluorimetric Gus assay was performed to detect quantitative Gus expression in the resistant calli. Resistant calli were grown under various conditions, e.g. 0.1% NaCl, or 2 mg/L of ABA in callus medium, or without stress (normal conditions). The level of promoter activity for both the 35S and Salt promoters was measured on the basis of Gus expression detected with fluorimetric assay. The results showed that the transformed calli (varieties Arval and White Martin) with plasmid pGVB310 showed Gus expression that was about 8-12 times higher than that of the transformed calli with plasmid p35SGus. 2 refs, 1 fig., 1 tab

  9. Metabolism of 35S- and 14C-labeled propylthiouracil in a model in vitro system containing thyroid peroxidase

    In previous communications we described an in vitro model system containing highly purified thyroid peroxidase (TPO) for studying the mechanism of inhibition of thyroid hormone biosynthesis by the antithyroid drugs, 6-propylthiouracil (PTU) and 1-methyl-2-mercaptoimidazole (MMI). We showed that inhibition of iodination of thyroglobulin in this system may be reversible or irreversible depending on the relative concentrations of iodide and drug and the TPO concentration. Metabolism of the drugs occurred under both conditions, but was more limited under irreversible conditions of inhibition. It was of interest to examine the nature of the drug metabolites associated with reversible and irreversible conditions of inhibition. For this purpose we have employed the 35S- and 14C-labeled drugs and a recently developed reverse phase HPLC procedure. Results of a similar study with MMI were reported in an earlier communication. In the present study we report our findings with PTU. Under conditions of reversible inhibition, PTU was readily metabolized and by 15 min was reduced to a few percent of the starting value. The earliest detectable metabolite with both [35S]- and [14C]PTU was the disulfide, which reached a peak in about 15 min and then slowly declined. Coincident with the decline in the disulfide was the appearance of more polar metabolites. In the case of [35S]PTU, these corresponded to sulfate/sulfite, PTU sulfonate, and a product tentatively identified as PTU sulfinate. The latter two were also observed as 14C-labeled metabolites produced from [14C]PTU. Two nonpolar desulfurated 14C-labeled metabolites were also observed. Surprisingly, these did not correspond to either propyluracil or propyldeoxyuracil, the anticipated most likely products of PTU desulfuration. The identity of these desulfurated metabolites of PTU in the TPO model system remains to be determined

  10. Location, degree, and direction of DNA bending associated with the Hin recombinational enhancer sequence and Fis-enhancer complex.

    Perkins-Balding, D; D. P Dias; Glasgow, A. C.

    1997-01-01

    The Fis protein of Escherichia coli and Salmonella typhimurium stimulates several site-specific DNA recombination reactions, as well as transcription of a number of genes. Fis binds to a 15-bp core recognition sequence and induces DNA bending. Mutations in Fis which alter its ability to bend DNA have been shown to reduce the stimulatory activity of Fis in both site-specific recombination and transcription systems. To examine the role of DNA bending in the activity of the Fis-recombinational e...

  11. Quantitative autoradiographic investigations on hairs, skin, and nails with the precursors 35S-cystine or 35S-methionine and 3H-thymidine respectively in animal experiments

    By quantitative autoradiographic methods we tested the possible influence of the sulphurous amino acid L-cystine on the growth of hair, skin and nails. In the 3H-thymidine autoradiographic method the tracing index (percentage of traced cell cores in relation to the total number of cell cores of a cell population) is calculated morphologically. In the 35S-cystine and the 35S-methionine autoradiography a quantification is carried out by applying the microscopic photometer for the investigation of defined hair root cross-sections. Measured values are indicated which depend on dosage and on the incorporation time. The investigation of other agents provoking negative or positive reactions of the hair growth by means of this method will have to be realized. Unlabelled L-cystine seems to promote the growth of hairs, epidermic basal cells and of germ cells of mouse nails, according to experiences made in animal experiments. The latter findings will have to be completed and confirmed by additional experiments. (orig.)

  12. Prospective Whole-Genome Sequencing Enhances National Surveillance of Listeria monocytogenes

    Kwong, Jason C.; Mercoulia, Karolina; Tomita, Takehiro; Easton, Marion; Li, Hua Y.; Bulach, Dieter M.; Stinear, Timothy P.; Seemann, Torsten; Benjamin P Howden

    2016-01-01

    Whole-genome sequencing (WGS) has emerged as a powerful tool for comparing bacterial isolates in outbreak detection and investigation. Here we demonstrate that WGS performed prospectively for national epidemiologic surveillance of Listeria monocytogenes has the capacity to be superior to our current approaches using pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), multilocus variable-number tandem-repeat analysis (MLVA), binary typing, and serotyping. Initially 423 ...

  13. Daytime sleep enhances consolidation of the spatial but not motoric representation of motor sequence memory.

    Geneviève Albouy

    Full Text Available Motor sequence learning is known to rely on more than a single process. As the skill develops with practice, two different representations of the sequence are formed: a goal representation built under spatial allocentric coordinates and a movement representation mediated through egocentric motor coordinates. This study aimed to explore the influence of daytime sleep (nap on consolidation of these two representations. Through the manipulation of an explicit finger sequence learning task and a transfer protocol, we show that both allocentric (spatial and egocentric (motor representations of the sequence can be isolated after initial training. Our results also demonstrate that nap favors the emergence of offline gains in performance for the allocentric, but not the egocentric representation, even after accounting for fatigue effects. Furthermore, sleep-dependent gains in performance observed for the allocentric representation are correlated with spindle density during non-rapid eye movement (NREM sleep of the post-training nap. In contrast, performance on the egocentric representation is only maintained, but not improved, regardless of the sleep/wake condition. These results suggest that motor sequence memory acquisition and consolidation involve distinct mechanisms that rely on sleep (and specifically, spindle or simple passage of time, depending respectively on whether the sequence is performed under allocentric or egocentric coordinates.

  14. Source of error in the chromatographic study of 35S-sulfate labeled mucous glycoproteins secreted by the gill epithelium of Mytilus edulis

    HPLC combined with [35S]-sulfate/[3H]-glucosamine radiolabeling were employed to study the synthesis and secretion of mucous glycoproteins. The secreted radiolabeled glycoproteins were separated from the medium by precipitation with a mixture of trichloroacetic-phosphotungstic acids (TCA/PTA). The redissolved glycoproteins were chromatographed on an anion exchange protein column at varying pH of the mobile phase and fractions were collected for liquid scintillation counting. Varying the pH of the mobile phase from pH 3 to 7 resulted in a decrease of glycoprotein bound [35S] from 69.5 to 0.5% of the total recovered [35S]-sulfate with the remainder recovered as free [35S]-sulfate. The [3H]-labeled glycoprotein recovered under the uV peaks at this pH range was 99.5%. When high performance size exclusion chromatography was performed the change in mobile phase pH did not affect the 100% recovery of either [35S]-or [3H]-labels under the uV peaks. No free [35S]-sulfate was obtained when [35S]-labeled glycoproteins were separated from the medium using dialysis. These data suggest that the standard method of TCA/PTA precipitation of [35S]-labeled glycoproteins may cleave the [35S]-sulfate ester linkages to the oligosaccharide chains. The [35S]-sulfate may then rebind to the macromolecule by a relatively strong noncovalent bond. This may prove critical in anion exchange protein HPLC studies

  15. (/sup 35/S)autoradiographic study of sulfated GAG accumulation and turnover in embryonic mouse tooth germs

    Lau, E.C.; Boukari, A.; Arechaga, J.; Osman, M.; Ruch, J.V.

    1983-01-01

    The accumulation of sulfated glycosaminoglycans(GAG) in embryonic mouse molars before, during, and after terminal differentiation of odontoblasts was localized by (/sup 35/S)autoradiography combined with the use of chondroitin ABC lyase. Much more sulfated GAG were accumulated in the dental papilla than in the dental epithelium. High incorporation of (/sup 35/S)sulfate occurred at the epithelio-mesenchymal junction, which is the site of dental basement membrane and predentin. Before terminal differentiation of odontoblasts, the distribution of sulfated GAG was uniform at the basement membrane. After the onset of terminal differentiation of odontoblasts, much more sulfated GAG accumulated at the tip of principal cusps than at the apical (inferior) parts of cusps, and sulfated GAG were then found to be degraded more rapidly at the epithelio-mesenchymal junction than at other parts of the tooth germ. Thus regional variation in the rate of degradation of GAG exists in the tooth germs. Trypsin-isolated dental epithelia cultured in vitro synthesized a new basement membrane that could be labeled with (/sup 3/H)glucosamine but not with /sup 35/SO4(-2). The epithelial-derived basal lamina contains little or no sulfatated GAG.

  16. Enhancement of aerobic granulation by zero-valent iron in sequencing batch airlift reactor

    Highlights: • Zero-valent iron (ZVI) was used firstly to enhance the aerobic granulation. • ZVI significantly decreased the start-up time of the aerobic granulation. • ZVI had the function of enhancing organic material diversity identified by 3-D EEM. • ZVI could enhance the diversity of microbial community. - Abstract: This study elucidates the enhancement of aerobic granulation by zero-valent iron (ZVI). A reactor augmented with ZVI had a start-up time of aerobic granulation (43 days) that was notably less than that for a reactor without augmentation (64 days). The former reactor also had better removal efficiencies for chemical oxygen demand and ammonium. Moreover, the mature granules augmented with ZVI had better physical characteristics and produced more extracellular polymeric substances (especially of protein). Three-dimensional-excitation emission matrix fluorescence showed that ZVI enhanced organic material diversity. Additionally, ZVI enhanced the diversity of the microbial community. Fe2+ dissolution from ZVI helped reduce the start-up time of aerobic granulation and increased the extracellular polymeric substance content. Conclusively, the use of ZVI effectively enhanced aerobic granulation

  17. Enhancement of aerobic granulation by zero-valent iron in sequencing batch airlift reactor

    Kong, Qiang, E-mail: kongqiang0531@hotmail.com [College of Life Science, Shandong Normal University, 88 Wenhua Donglu, Jinan 250014, Shandong (China); Ngo, Huu Hao [School of Civil and Environmental Engineering, University of Technology Sydney, Broadway, NSW 2007 (Australia); Shu, Li [School of Engineering, Faculty of Science, Engineering and Built Environment, Deakin University, Geelong, Victoria 3216 (Australia); Fu, Rong-shu; Jiang, Chun-hui [College of Life Science, Shandong Normal University, 88 Wenhua Donglu, Jinan 250014, Shandong (China); Miao, Ming-sheng, E-mail: mingshengmiao@163.com [College of Life Science, Shandong Normal University, 88 Wenhua Donglu, Jinan 250014, Shandong (China)

    2014-08-30

    Highlights: • Zero-valent iron (ZVI) was used firstly to enhance the aerobic granulation. • ZVI significantly decreased the start-up time of the aerobic granulation. • ZVI had the function of enhancing organic material diversity identified by 3-D EEM. • ZVI could enhance the diversity of microbial community. - Abstract: This study elucidates the enhancement of aerobic granulation by zero-valent iron (ZVI). A reactor augmented with ZVI had a start-up time of aerobic granulation (43 days) that was notably less than that for a reactor without augmentation (64 days). The former reactor also had better removal efficiencies for chemical oxygen demand and ammonium. Moreover, the mature granules augmented with ZVI had better physical characteristics and produced more extracellular polymeric substances (especially of protein). Three-dimensional-excitation emission matrix fluorescence showed that ZVI enhanced organic material diversity. Additionally, ZVI enhanced the diversity of the microbial community. Fe{sup 2+} dissolution from ZVI helped reduce the start-up time of aerobic granulation and increased the extracellular polymeric substance content. Conclusively, the use of ZVI effectively enhanced aerobic granulation.

  18. Preparation and high-performance liquid chromatography of 3'-phosphoadenosine-5'-phospho[35S]sulfate with a predetermined specific activity

    3'-Phosphoadenosine-5'-phospho[35S]sulfate (PAP35S) was prepared by incubating ATP and carrier-free H2(35)SO4 with a 100,000g supernatant fraction prepared from chick embryo chondrocytes. The product was partially purified by paper electrophoresis and mixed with unlabeled PAPS to give a solution of PAP35S with a specific activity and a concentration approximating those required for the desired metabolic studies. The product was analyzed by high-performance liquid chromatography on an anion-exchange column to determine the proportion of the 35SO4 cpm and A260 material found in the PAPS and other contaminating nucleotides. The PAP35S was purified further by preparative high-performance liquid chromatography. The exact specific activity of the PAP35S was then determined by using this PAP35S preparation as the SO4 donor in a sulfotransferase reaction using a microsomal preparation from the chick embryo chondrocytes as the enzyme and an 3H-labeled oligosaccharide as the SO4 acceptor. The sulfated oligosaccharide was then isolated and the number of 3H and 35SO4 counts per minute in this product were used to calculate the specific activity of the donor. The features of this generally useful approach for preparing PAP35S of any desired specific activity and concentration are discussed

  19. RNA-guided complex from a bacterial immune system enhances target recognition through seed sequence interactions

    Wiedenheft, Blake; van Duijn, Esther; Bultema, Jelle; Waghmare, Sakharam; Zhou, Kaihong; Barendregt, Arjan; Westphal, Wiebke; Heck, Albert; Boekema, Egbert; Dickman, Mark; Doudna, Jennifer A.

    2011-01-01

    Prokaryotes have evolved multiple versions of an RNA-guided adaptive immune system that targets foreign nucleic acids. In each case, transcripts derived from clustered regularly interspaced short palindromic repeats (CRISPRs) are thought to selectively target invading phage and plasmids in a sequenc

  20. A Monte Carlo-based framework enhances the discovery and interpretation of regulatory sequence motifs

    Seitzer Phillip

    2012-11-01

    Full Text Available Abstract Background Discovery of functionally significant short, statistically overrepresented subsequence patterns (motifs in a set of sequences is a challenging problem in bioinformatics. Oftentimes, not all sequences in the set contain a motif. These non-motif-containing sequences complicate the algorithmic discovery of motifs. Filtering the non-motif-containing sequences from the larger set of sequences while simultaneously determining the identity of the motif is, therefore, desirable and a non-trivial problem in motif discovery research. Results We describe MotifCatcher, a framework that extends the sensitivity of existing motif-finding tools by employing random sampling to effectively remove non-motif-containing sequences from the motif search. We developed two implementations of our algorithm; each built around a commonly used motif-finding tool, and applied our algorithm to three diverse chromatin immunoprecipitation (ChIP data sets. In each case, the motif finder with the MotifCatcher extension demonstrated improved sensitivity over the motif finder alone. Our approach organizes candidate functionally significant discovered motifs into a tree, which allowed us to make additional insights. In all cases, we were able to support our findings with experimental work from the literature. Conclusions Our framework demonstrates that additional processing at the sequence entry level can significantly improve the performance of existing motif-finding tools. For each biological data set tested, we were able to propose novel biological hypotheses supported by experimental work from the literature. Specifically, in Escherichia coli, we suggested binding site motifs for 6 non-traditional LexA protein binding sites; in Saccharomyces cerevisiae, we hypothesize 2 disparate mechanisms for novel binding sites of the Cse4p protein; and in Halobacterium sp. NRC-1, we discoverd subtle differences in a general transcription factor (GTF binding site motif

  1. Enhancement of aerobic granulation by zero-valent iron in sequencing batch airlift reactor.

    Kong, Qiang; Ngo, Huu Hao; Shu, Li; Fu, Rong-Shu; Jiang, Chun-Hui; Miao, Ming-sheng

    2014-08-30

    This study elucidates the enhancement of aerobic granulation by zero-valent iron (ZVI). A reactor augmented with ZVI had a start-up time of aerobic granulation (43 days) that was notably less than that for a reactor without augmentation (64 days). The former reactor also had better removal efficiencies for chemical oxygen demand and ammonium. Moreover, the mature granules augmented with ZVI had better physical characteristics and produced more extracellular polymeric substances (especially of protein). Three-dimensional-excitation emission matrix fluorescence showed that ZVI enhanced organic material diversity. Additionally, ZVI enhanced the diversity of the microbial community. Fe(2+) dissolution from ZVI helped reduce the start-up time of aerobic granulation and increased the extracellular polymeric substance content. Conclusively, the use of ZVI effectively enhanced aerobic granulation. PMID:25108827

  2. Endoplasmic reticulum targeting sequence enhances HBV-specific cytotoxic T lymphocytes induced by a CTL epitope-based DNA vaccine

    CD8+ T cells play a critical role in protective immunity against Hepatitis B Virus (HBV). Epitope-based DNA vaccines expressing HBV-dominant CTL epitopes can be used as candidate vaccines capable of inducing cytotoxic T Lymphocytes (CTL) responses. A plasmid DNA encoding a CTL epitope of HBV core antigen, HBc18-27, was constructed. Intramuscular immunization of C57BL/6 mice with this DNA vaccine resulted in successful induction of HBV-specific CTL responses. In order to promote transportation of the peptide into endoplasmic reticulum (ER) to bind to MHC class I molecules for optimal class I antigen presentation, an ER targeting sequence (ERTS) was fused with the C18-27 encoding gene. ERTS fusion significantly enhanced specific CD8+ T cell responses in terms of CTL cytolysis as well as IFN-γ secretion. This enhancement was correlated with promoted epitope presentation on target cell surface. We report here an enhanced immunogenicity of an epitope-based DNA vaccine using an ER targeting signal sequence, which has significant implications for future design of therapeutic HBV vaccine

  3. (14)N overtone NMR under MAS: signal enhancement using symmetry-based sequences and novel simulation strategies.

    Haies, Ibraheem M; Jarvis, James A; Bentley, Harry; Heinmaa, Ivo; Kuprov, Ilya; Williamson, Philip T F; Carravetta, Marina

    2015-03-01

    Overtone (14)N NMR spectroscopy is a promising route for the direct detection of (14)N signals with good spectral resolution. Its application is currently limited, however, by the absence of efficient polarization techniques for overtone signal enhancement and the lack of efficient numerical simulation techniques to aid in both the development of new methods and the analysis and interpretation of experimental data. In this paper we report a novel method for the transfer of polarization from (1)H to the (14)N overtone using symmetry-based R-sequences that overcome many of the limitations of adiabatic approaches that have worked successfully on static samples. Refinement of these sequences and the analysis of the resulting spectra have been facilitated through the development of an efficient simulation strategy for (14)N overtone NMR spectroscopy of spinning samples, using effective Hamiltonians on top of Floquet and Fokker-Planck equations. PMID:25662410

  4. Nucleotide sequence analysis and enhancer function of long terminal repeats associated with an endogenous African green monkey retroviral DNA.

    Kessel, M; Khan, A S

    1985-01-01

    The nucleotide sequence and enhancer activity of the long terminal repeats (LTRs) associated with a cloned endogenous African green monkey (AGM) retroviral DNA designated as lambda-AGM-1 was studied. A unique feature of the endogenous AGM proviral LTRs was the presence of multiple copies of two types of directly repeating units in the U3 region: 16 8-base-pair (bp) repeats were present in the 5' LTR and 12 were present in the 3' LTR which were bound by a 6-bp perfect direct repeat; tandem dup...

  5. Measurement of the Forward-Backward Asymmetry Coefficient in the $^{35}(n, p)^{35}S$ Reaction for Resonance Neutrons

    Vesna, V A; Okunev, I S; Oprya, A; Salatskii, V I; Sedyshev, P V; Shalanski, P I

    2002-01-01

    The measurements of the P-even forward-backward correlation in the ^{35}(n, p)^{35}S reaction were carried out at resonance unpolarized neutrons of the IBR-30 pulsed facility. The alpha^{FB} values have been determined for neutron energy regions at averaged values of 3.1, 195 and 398 eV: (5.3\\pm 7.6)\\cdot 10^{-3}, (1.68\\pm 0.50)\\cdot 10^{-1}, and -(7.4\\pm 3.6)\\cdot 10^{-3}, respectively. Involving results of the experiment at thermal polarized neutrons on measurement of the P-odd and the P-even left-right correlations in given reaction, the weak matrix element has been estimated, and an analysis of the possible values of the neutron and proton amplitudes for channels with total momentum j=1/2 and 3/2 has been performed.

  6. Control of gene conversion and somatic hypermutation by immunoglobulin promoter and enhancer sequences

    Yang, Shu Yuan; Fugmann, Sebastian D.; Schatz, David G.

    2006-01-01

    It is thought that gene conversion (GCV) and somatic hypermutation (SHM) of immunoglobulin (Ig) genes occur in two steps: the generation of uracils in DNA by activation-induced cytidine deaminase, followed by their subsequent repair by various DNA repair pathways to generate sequence-diversified products. It is not known how either of the two steps is targeted specifically to Ig loci. Because of the tight link between transcription and SHM, we have investigated the role of endogenous Ig light...

  7. Bioaggregate of photo-fermentative bacteria for enhancing continuous hydrogen production in a sequencing batch photobioreactor

    Guo-Jun Xie; Bing-Feng Liu; Rui-Qing Wang; Jie Ding; Hong-Yu Ren; Xu Zhou; Nan-Qi Ren

    2015-01-01

    Hydrogen recovery through solar-driven biomass conversion by photo-fermentative bacteria (PFB) has been regarded as a promising way for sustainable energy production. However, a considerable fraction of organic substrate was consumed for the growth of PFB as biocatalysts, furthermore, these PFB were continuously washed out from the photobioreactor in continuous operation because of their poor flocculation. In this work, PFB bioaggregate induced by L-cysteine was applied in a sequencing batch ...

  8. Drosophila ARSs contain the yeast ARS consensus sequence and a replication enhancer.

    Mills, J S; Kingsman, A J; Kingsman, S M

    1986-01-01

    A number of restriction fragments that function as autonomously replicating sequences (ARSs) in yeast have been isolated from Drosophila melanogaster DNA. The behaviour in yeast of plasmids containing Drosophila ARS elements was studied and compared to that exhibited by the archetypal yeast ARS-1 plasmid. ARS functions were localised by subcloning and BAL-31 deletion analysis. These studies demonstrated the structural and functional complexity of Drosophila ARSs. Each Drosophila ARS element h...

  9. A noncontrast-enhanced pulse sequence optimized to visualize human peripheral vessels

    Gjesdal, Kjell-Inge [Sunnmoere MR-Klinikk, Aalesund (Norway); Storaas, Tryggve [Ullevaal University Hospital, Section for Diagnostic Physics, Department of Radiology, Oslo (Norway); Geitung, Jonn-Terje [Haraldsplass University Hospital, Department of Radiology, Bergen (Norway)

    2009-01-15

    The purpose of this paper is to present a pulse sequence optimized to visualize human peripheral vessels. The optimized MR technique is a 3D multi-shot balanced non-SSFP gradient echo pulse sequence with fat suppression. Several imaging parameters were adjusted to find the best compromise between the contrast of vascular structures and muscle, fat, and bone. Most of the optimization was performed in the knee and calf regions using multi-channel SENSE coils. To verify potential clinical use, images of both healthy volunteers and volunteers with varicose veins were produced. The balanced non-SSFP sequence can produce high-spatial-resolution images of the human peripheral vessels without the need for an intravenous contrast agent. Both arteries and veins are displayed along with other body fluids. Due to the high spatial resolution of the axial plane source or reconstructed images, the need for procedures to separate arteries from veins is limited. We demonstrate that high signals from synovial joint fluid and cystic structures can be suppressed by applying an inversion prepulse but at the expense of reduced image signal-to-noise and overall image quality. (orig.)

  10. A noncontrast-enhanced pulse sequence optimized to visualize human peripheral vessels

    The purpose of this paper is to present a pulse sequence optimized to visualize human peripheral vessels. The optimized MR technique is a 3D multi-shot balanced non-SSFP gradient echo pulse sequence with fat suppression. Several imaging parameters were adjusted to find the best compromise between the contrast of vascular structures and muscle, fat, and bone. Most of the optimization was performed in the knee and calf regions using multi-channel SENSE coils. To verify potential clinical use, images of both healthy volunteers and volunteers with varicose veins were produced. The balanced non-SSFP sequence can produce high-spatial-resolution images of the human peripheral vessels without the need for an intravenous contrast agent. Both arteries and veins are displayed along with other body fluids. Due to the high spatial resolution of the axial plane source or reconstructed images, the need for procedures to separate arteries from veins is limited. We demonstrate that high signals from synovial joint fluid and cystic structures can be suppressed by applying an inversion prepulse but at the expense of reduced image signal-to-noise and overall image quality. (orig.)

  11. Detection of GAD-Ab index in diabetic patients using 35S labeled recombinant human GAD65 antigen

    Objective: To establish a novel method for measuring glutamic acid decarboxylase autoanti-bodies(GAD-Ab). Methods: Recombinant human GAD65 was used as the antigen, in vitro transcribed and translated 35S-GAD65 as the tracer, a self-designed rotating incubation apparatus as the incubator, protein-A sepharose as the precipitator, and the liquid scintillation counter was used to measure radioactive count value to detect GAD-Ab. The positive cut-off point of GAD-Ab index was determined as > 0.05 by the 99.5% percentile in 109 healthy individuals. GAD-Ab levels were determined in 43 type 1 and 226 type 2 diabetic patients. Results: The optimized working conditions included SJ1515 35S-methionine for in vitro transcription and translation, 20-30 r/min setup of rotating incubation apparatus, test temperature 4-25 degree C, freshly prepared buffer of pH 7.2-7.4, and horizontal rotor centrifuge. The new method was better than original one, with intra-assay CV of 4.9%-8.3% and inter-assay CV of 7.1%-10.8 %, specificity of 98.2%. The results were comparable with the figures issued by an international standardized laboratory (concordance was 98.3%, Kappa value 0.971). The positive rate of GAD-Ab was 58.1% (25 of 43) in type 1 and 10.2%(23 of 226) in type 2 diabetes patients, but only 1.8% (2 of 109) in healthy individuals. Conclusion: The new assay for GAD-Ab is a highly sensitive, accurate, specific and reproducible method for clinical use

  12. Investigating and correcting plasma DNA sequencing coverage bias to enhance aneuploidy discovery.

    Dineika Chandrananda

    Full Text Available Pregnant women carry a mixture of cell-free DNA fragments from self and fetus (non-self in their circulation. In recent years multiple independent studies have demonstrated the ability to detect fetal trisomies such as trisomy 21, the cause of Down syndrome, by Next-Generation Sequencing of maternal plasma. The current clinical tests based on this approach show very high sensitivity and specificity, although as yet they have not become the standard diagnostic test. Here we describe improvements to the analysis of the sequencing data by reducing GC bias and better handling of the genomic repeats. We show substantial improvements in the sensitivity of the standard trisomy 21 statistical tests, which we measure by artificially reducing read coverage. We also explore the bias stemming from the natural cleavage of plasma DNA by examining DNA motifs and position specific base distributions. We propose a model to correct this fragmentation bias and observe that incorporating this bias does not lead to any further improvements in the detection of fetal trisomy. The improved bias corrections that we demonstrate in this work can be readily adopted into existing fetal trisomy detection protocols and should also lead to improvements in sub-chromosomal copy number variation detection.

  13. Prospective Whole-Genome Sequencing Enhances National Surveillance of Listeria monocytogenes.

    Kwong, Jason C; Mercoulia, Karolina; Tomita, Takehiro; Easton, Marion; Li, Hua Y; Bulach, Dieter M; Stinear, Timothy P; Seemann, Torsten; Howden, Benjamin P

    2016-02-01

    Whole-genome sequencing (WGS) has emerged as a powerful tool for comparing bacterial isolates in outbreak detection and investigation. Here we demonstrate that WGS performed prospectively for national epidemiologic surveillance of Listeria monocytogenes has the capacity to be superior to our current approaches using pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), multilocus variable-number tandem-repeat analysis (MLVA), binary typing, and serotyping. Initially 423 L. monocytogenes isolates underwent WGS, and comparisons uncovered a diverse genetic population structure derived from three distinct lineages. MLST, binary typing, and serotyping results inferred in silico from the WGS data were highly concordant (>99%) with laboratory typing performed in parallel. However, WGS was able to identify distinct nested clusters within groups of isolates that were otherwise indistinguishable using our current typing methods. Routine WGS was then used for prospective epidemiologic surveillance on a further 97 L. monocytogenes isolates over a 12-month period, which provided a greater level of discrimination than that of conventional typing for inferring linkage to point source outbreaks. A risk-based alert system based on WGS similarity was used to inform epidemiologists required to act on the data. Our experience shows that WGS can be adopted for prospective L. monocytogenes surveillance and investigated for other pathogens relevant to public health. PMID:26607978

  14. Short synthetic oligodeoxyribonucleotide leader sequences enhance accumulation of human proinsulin synthesized in Escherichia coli.

    Sung, W. L.; Yao, F L; Zahab, D M; Narang, S A

    1986-01-01

    Enhanced accumulation of human proinsulin synthesized in Escherichia coli has been achieved by inserting a short leader of homooligopeptide at the amino end of proinsulin. Out of 20 amino acid oligomers studied, (Ala)6, (Asn)6, (Cys)7, (Gln)7, (His)6, (Ser)6, and (Thr)6 leaders were the most effective, with the yield of proinsulin ranging between 6% and 26% of the total bacterial protein. These constructions were made by inserting a synthetic oligodeoxyribonucleotide duplex, coding for a smal...

  15. Instability strips of main sequence B stars: a parametric study of iron enhancement

    Miglio, A; Montalban, J; Dupret, M A

    2006-01-01

    The discovery of beta Cephei stars in low metallicity environments, as well as the difficulty to theoretically explain the excitation of the pulsation modes observed in some beta Cephei and SPB stars, suggest that the iron opacity ``bump'' provided by standard models could be underestimated. We investigate, by means of a parametric study, the effect of a local iron enhancement on the location of the beta Cephei and SPB instability strips.

  16. Enhanced DNA sequencing performance through edge-hydrogenation of graphene electrodes

    He, Yuhui; Grigoriev, Anton; Ahuja, Rajeev; Long, Shibing; Huo, ZongLiang; Liu, Ming

    2010-01-01

    We propose using graphene electrodes with hydrogenated edges for solid-state nanopore-based DNA sequencing, and perform molecular dynamics simulations in conjunction with electronic transport calculations to explore the potential merits of this idea. The results of our investigation show that, compared to the unhydrogenated system, edge-hydrogenated graphene electrodes facilitate the temporary formation of H-bonds with suitable atomic sites in the translocating DNA molecule. As a consequence, the average conductivity is drastically raised by about 3 orders of magnitude while exhibiting significantly reduced statistical variance. We have furthermore investigated how these results are affected when the distance between opposing electrodes is varied and have identified two regimes: for narrow electrode separation, the mere hindrance due to the presence of protruding hydrogen atoms in the nanopore is deemed more important, while for wider electrode separation, the formation of H-bonds becomes the dominant effect....

  17. Analysis of DNA sequences which regulate the transcription of herpes simplex virus immediate early gene 3: DNA sequences required for enhancer-like activity and response to trans-activation by a virion polypeptide.

    Bzik, D J; Preston, C.M.

    1986-01-01

    The far upstream region of herpes simplex virus (HSV) immediate early (IE) gene 3 has previously been shown to increase gene expression in an enhancer-like manner, and to contain sequences which respond to stimulation of transcription by a virion polypeptide, Vmw65. To analyse the specific DNA sequences which mediate these functions, sequential deletions from each end of the far upstream region were made. The effects of the deletions on transcription in the absence or presence of the Vmw65 we...

  18. Studies on distribution and residue of sulfur in simulated acid rain in vegetable and soil by using 35S

    Distribution and residue of sulfur in simulated acid rain in two kinds of vegetables (lettuce and Chinese cabbage) and three types of soils (acid yellow earth, acid and neutral purple soils) were studied by using 35S tracer method. The results showed that the higher concentration of acid rain was sprayed, the more residue of sulfur in vegetable there would be. The residue of sulfur in vegetable varied with the different physical and chemical properties of soils, the order of sulfur residue in vegetable was: acid purple soil>acid yellow earth>neutral purple soil. In the same soil, the residue of sulfur in lettuce was higher than that in Chinese cabbage, for the same vegetable, the residue of sulfur in leaves were higher than that in stems. The order of sulfur residue in different soils was acid purple soil>acid yellow earth>neutral purple soil. The higher concentration of acid rain was sprayed, the more residue of sulfur in soil surface there would be. The sulfur residue varied with the depth of soil and the pH value of acid rain. With the increase of soil depth, a slight increase of sulfur residue with rain of ph 6 and a slight decrease with rain of pH 4.0 and 2.5 were found

  19. Complete genome sequence of virulence-enhancing Siphophage VHS1 from Vibrio harveyi.

    Khemayan, Krit; Prachumwat, Anuphap; Sonthayanon, Burachai; Intaraprasong, Aungkul; Sriurairatana, Siriporn; Flegel, Timothy W

    2012-04-01

    Vibrio harveyi siphophage 1 (VHS1) is a tailed phage with an icosahedral head of approximately 66 nm in diameter and an unornamented, flexible tail of approximately 153 nm in length. When Vibrio harveyi 1114GL is lysogenized with VHS1, its virulence for the black tiger shrimp (Penaeus monodon) increases by more than 100 times, and this coincides with production of a toxin(s) associated with shrimp hemocyte agglutination. Curiously, the lysogen does not show increased virulence for the whiteleg shrimp (Penaeus [Litopenaeus] vannamei). Here we present and annotate the complete, circular genome of VHS1 (81,509 kbp; GenBank accession number JF713456). By software analysis, the genome contains 125 putative open reading frames (ORFs), all of which appear to be located on the same DNA strand, similar to the case for many other bacteriophages. Most of the putative ORFs show no significant homology to known sequences in GenBank. Notable exceptions are ORFs for a putative DNA polymerase and putative phage structural proteins, including a portal protein, a phage tail tape measure protein, and a phage head protein. The last protein was identified as a component of the species-specific toxin mixture described above as being associated with agglutination of hemocytes from P. monodon. PMID:22307287

  20. Superselective Particle Embolization Enhances Efficacy of Radiofrequency Ablation: Effects of Particle Size and Sequence of Action

    Purpose. To evaluate the effects of particle size and course of action of superselective bland transcatheter arterial embolization (TAE) on the efficacy of radiofrequency ablation (RFA). Methods. Twenty pigs were divided into five groups: group 1a, 40-μm bland TAE before RFA; group 1b, 40-μm bland TAE after RFA; group 2a, 250-μm bland TAE before RFA; group 2b, 250-μm bland TAE after RFA and group 3, RFA alone. A total of 40 treatments were performed with a combined CT and angiography system. The sizes of the treated zones were measured from contrast-enhanced CTs on days 1 and 28. Animals were humanely killed, and the treated zones were examined pathologically. Results. There were no complications during procedures and follow-up. The short-axis diameter of the ablation zone in group 1a (mean ± standard deviation, 3.19 ± 0.39 cm) was significantly larger than in group 1b (2.44 ± 0.52 cm; P = 0.021), group 2a (2.51 ± 0.32 cm; P = 0.048), group 2b (2.19 ± 0.44 cm; P = 0.02), and group 3 (1.91 ± 0.55 cm; P 3). At histology, 40-μm microspheres were observed to occlude smaller and more distal arteries than 250-μm microspheres. Conclusion. Bland TAE is more effective before RFA than postablation embolization. The use of very small 40-μm microspheres enhances the efficacy of RFA more than the use of larger particles.

  1. MRI evaluation of myometrial invasion by endometrial carcinoma. Comparison between fast-spin-echo T2W and coronal FMPSPGR Gadolinium-Dota-Enhanced Sequences

    Purpose. The depth of myometrial invasion by endometrial carcinoma strongly affects the incidence of metastasis to regional nodes and influences the surgical strategies. The aim of this paper is to compare the results of FSE T2-w and Gadolinium-enhanced FMPSGR MR sequences in assessing the depth of myometrial invasion by endometrial cancer. Materials and methods. Forty-five women with histopathologically-proven endometrial carcinoma underwent preoperative MRI. Axial SE TI w, axial, sagittal and para-coronal FSE T2w and para-coronal Gadolinium enhanced FMPSGR sequences were performed using a high field strength magnet (1.5T). Within one month of MR all patients underwent hysterectomy, and anatomical evaluation of the surgical specimen was done sectioning the uterus along the short axis. Based upon the results of the histological evaluation the results of the FSE T2w and Gadolinium-enhanced sequences were compared and the statistical difference between the results obtained was statistically evaluated. Results. The histological evaluation showed intra mucosal neoplasm in 11 patients, myometrial infiltration less than 50% in 31 patients, myometrial infiltration more than 50% in 12 patients and transmural cancer 1 patient. Statistical evaluation showed that the FSE T2w sequence had a global sensitivity and specificity of 80.6% and 87.6%, respectively, with a mean Negative Predictive Value of 92.6% and a mean Positive Predictive Value of 86%. Gadolinium-enhanced FMPSPGR sequence had a global sensitivity and specificity of 90.6% and 93.3%, respectively, with a mean Negative Predictive Value of 96,3% and a mean Positive Predictive Value of 88%. The staging accuracy (χ2 test) on FMPSPGR images (95%) was higher than that on FSE T2w images (78%). Conclusions. In our experience Gadolinium-enhanced dynamic sequences increase the accuracy of MR imaging in diagnosing the depth of myometrial invasion. In particular they improve the visualisation of the inner myometrium, the so

  2. CRISPR/Cas9 system as an innovative genetic engineering tool: Enhancements in sequence specificity and delivery methods.

    Jo, Young-Il; Suresh, Bharathi; Kim, Hyongbum; Ramakrishna, Suresh

    2015-12-01

    While human gene therapy has gained significant attention for its therapeutic promise, CRISPR/Cas9 technology has made a breakthrough as an efficient genome editing tool by emulating prokaryotic immune defense mechanisms. Although many studies have found that CRISPR/Cas9 technology is more efficient, specific and manipulable than previous generations of gene editing tools, it can be further improved by elevating its overall efficiency in a higher frequency of genome modifications and reducing its off-target effects. Here, we review the development of CRISPR/Cas9 technology, focusing on enhancement of its sequence specificity, reduction of off-target effects and delivery systems. Moreover, we describe recent successful applications of CRISPR/Cas9 technology in laboratory and clinical studies. PMID:26434948

  3. Athletic injuries of the extensor carpi ulnaris subsheath: MRI findings and utility of gadolinium-enhanced fat-saturated T1-weighted sequences with wrist pronation and supination

    To report the magnetic resonance imaging (MRI) findings in athletic injuries of the extensor carpi ulnaris (ECU) subsheath, assessing the utility of gadolinium-enhanced (Gd) fat-saturated (FS) T1-weighted sequences with wrist pronation and supination. Sixteen patients (13 male, three female; mean age 30.3 years) with athletic injuries of the ECU subsheath sustained between January 2003 and June 2009 were included in this retrospective study. Initial and follow-up 1.5-T wrist MRIs were performed with transverse T1-weighted and STIR sequences in pronation, and Gd FS T1-weighted sequences with wrist pronation and supination. Two radiologists assessed the type of injury (A to C), ECU tendon stability, associated lesions and rated pulse sequences using a three-point scale: 1 = poor, 2 = good and 3 = excellent. Gd-enhanced FS T1-weighted transverse sequences in supination (2.63) and pronation (2.56) were most valuable, compared with STIR (2.19) and T1-weighted (1.94). Nine type A, one type B and six type C injuries were found. There were trends towards diminution in size, signal intensity and enhancement of associated pouches on follow-up MRI and tendon stabilisation within the ulnar groove. Gd-enhanced FS T1-weighted sequences with wrist pronation and supination are most valuable in assessing and follow-up athletic injuries of the ECU subsheath on 1.5-T MRI. (orig.)

  4. Athletic injuries of the extensor carpi ulnaris subsheath: MRI findings and utility of gadolinium-enhanced fat-saturated T1-weighted sequences with wrist pronation and supination

    Jeantroux, Jeremy; Guerini, Henri; Drape, Jean-Luc [Universite Paris Descartes, Department of Radiology B, Hopital Cochin, AP-HP, Paris (France); Becce, Fabio [Universite Paris Descartes, Department of Radiology B, Hopital Cochin, AP-HP, Paris (France); University of Lausanne, Department of Diagnostic and Interventional Radiology, Centre Hospitalier Universitaire Vaudois, Lausanne (Switzerland); Montalvan, Bernard [French Tennis Federation, Paris (France); Viet, Dominique Le [Hand Institute, Clinique Jouvenet, Paris (France)

    2011-01-15

    To report the magnetic resonance imaging (MRI) findings in athletic injuries of the extensor carpi ulnaris (ECU) subsheath, assessing the utility of gadolinium-enhanced (Gd) fat-saturated (FS) T1-weighted sequences with wrist pronation and supination. Sixteen patients (13 male, three female; mean age 30.3 years) with athletic injuries of the ECU subsheath sustained between January 2003 and June 2009 were included in this retrospective study. Initial and follow-up 1.5-T wrist MRIs were performed with transverse T1-weighted and STIR sequences in pronation, and Gd FS T1-weighted sequences with wrist pronation and supination. Two radiologists assessed the type of injury (A to C), ECU tendon stability, associated lesions and rated pulse sequences using a three-point scale: 1 = poor, 2 = good and 3 = excellent. Gd-enhanced FS T1-weighted transverse sequences in supination (2.63) and pronation (2.56) were most valuable, compared with STIR (2.19) and T1-weighted (1.94). Nine type A, one type B and six type C injuries were found. There were trends towards diminution in size, signal intensity and enhancement of associated pouches on follow-up MRI and tendon stabilisation within the ulnar groove. Gd-enhanced FS T1-weighted sequences with wrist pronation and supination are most valuable in assessing and follow-up athletic injuries of the ECU subsheath on 1.5-T MRI. (orig.)

  5. Strategy for complete NMR assignment of disordered proteins with highly repetitive sequences based on resolution-enhanced 5D experiments

    A strategy for complete backbone and side-chain resonance assignment of disordered proteins with highly repetitive sequence is presented. The protocol is based on three resolution-enhanced NMR experiments: 5D HN(CA)CONH provides sequential connectivity, 5D HabCabCONH is utilized to identify amino acid types, and 5D HC(CC-TOCSY)CONH is used to assign the side-chain resonances. The improved resolution was achieved by a combination of high dimensionality and long evolution times, allowed by non-uniform sampling in the indirect dimensions. Random distribution of the data points and Sparse Multidimensional Fourier Transform processing were used. Successful application of the assignment procedure to a particularly difficult protein, δ subunit of RNA polymerase from Bacillus subtilis, is shown to prove the efficiency of the strategy. The studied protein contains a disordered C-terminal region of 81 amino acids with a highly repetitive sequence. While the conventional assignment methods completely failed due to a very small differences in chemical shifts, the presented strategy provided a complete backbone and side-chain assignment.

  6. DELINEATION OF TECHNIQUES TO IMPLEMENT ON THE ENHANCED PROPOSED MODEL USING DATA MINING FOR PROTEIN SEQUENCE CLASSIFICATION

    Ananya Basu

    2014-02-01

    Full Text Available In post genomic era with the advent of new technologies a huge amount of complex molecular data are generated with high throughput. The management of this biological data is definitely a challenging task due to complexity and heterogeneity of data for discovering new knowledge. Issues like managing noisy and incomplete data are needed to be dealt with. Use of data mining in biological domain has made its inventory success. Discovering new knowledge from the biological data is a major challenge in data mining technique. The novelty of the proposed model is its combined use of intelligent techniques to classify the protein sequence faster and efficiently. Use of FFT, fuzzy classifier, String weighted algorithm, gram encoding method, neural network model and rough set classifier in a single model and in an appropriate place can enhance the quality of the classification system .Thus the primary challenge is to identify and classify the large protein sequences in a very fast and easy but intellectual way to decrease the time complexity and space complexity

  7. Sequence, taste and umami-enhancing effect of the peptides separated from soy sauce.

    Zhuang, Mingzhu; Lin, Lianzhu; Zhao, Mouming; Dong, Yi; Sun-Waterhouse, Dongxiao; Chen, Huiping; Qiu, Chaoying; Su, Guowan

    2016-09-01

    Five tasty peptides were separated from soy sauce, by sensory-guided fractionation, using macroporous resin, medium-pressure liquid chromatography and reverse phase-high performance liquid chromatography, and identified by ultra-performance liquid chromatography tandem mass-spectrometry as ALPEEV, LPEEV, AQALQAQA, EQQQQ and EAGIQ (which originated from glycinin A1bB2-445, glycinin A1bB2-445, cobyric acid synthase, leucine-tRNA ligase and glycoprotein glucosyltransferase, respectively). LPEEV, AQALQAQA and EQQQQ tasted umami with threshold values of 0.43, 1.25 and 0.76mmol/l, respectively. ALPEEV and EAGIQ had minimal umami taste, but ALPEEV, EAGIQ and LPEEV showed umami-enhancement with a threshold estimated at 1.52, 1.94 and 3.41mmol/l, respectively. In addition, the synthetic peptides showed much better sensory taste than mixtures of their constitutive amino acids. It indicated that peptides might play an important role in the umami taste of soy sauce. PMID:27041313

  8. Pharmacokinetics, tissue distribution, and cell localization of [35S]methionine-labeled recombinant human and murine alpha interferons in mice

    The pharmacokinetics, tissue distribution, cell localization, and penetration into tumor xenografts of recombinant [35S]methionine-labeled human alpha interferon (HuIFN-alpha) and murine alpha interferon (MuIFN-alpha) were examined in mice. Both interferons (IFNs) were removed from the blood in a rapid biphasic manner; HuIFN-alpha was cleared faster than MuIFN-alpha. Tissues were analyzed for radioactivity and over 90% of the IFNs was accounted for. The IFNs were detected predominantly in liver, kidney, gastrointestinal tract, pancreas, spleen, and lung. The levels of MuIFN-alpha compared with HuIFN-alpha were greater in the liver, spleen, and lung and less in the kidney, pancreas, and gastrointestinal tract. Heart, brain, testes, thymus, lymph nodes, fat, skin, and skeletal muscle contained much lower but measurable levels of both IFNs. There was penetration of HuIFN-alpha into tumor xenografts. The pharmacokinetics of IFN-alpha were independent of the strain of mouse, BALB/c or CBA, immune deprivation, or the presence of a tumor xenograft. Autoradiography of tissue sections from mice given injections of HuIFN-alpha or MuIFN-alpha indicated focal radioactivity in proximal convoluted tubules in the kidney and diffuse radioactivity in the liver, gastrointestinal tract, and pancrease. MuIFN-alpha, but not HuIFN-alpha, showed intense localization in cells in hepatic sinusoids, marginal zones in the spleen, and pulmonary alveolar walls, suggesting uptake by cells of the monocyte/macrophage lineage in these sites. The study shows the utility of biosynthetic labeling for pharmacokinetic studies of cytokines, clear differences in tissue distribution of IFN-alpha according to its species of origin, and targeting of homologous IFN-alpha to cells of the monocytic lineage

  9. Distribution of 35S-labelled BCG after application to the camera oculi anterior of BCG vaccinated and non-vaccinated rabbits

    After application of 35S-labelled BCG to the camera oculi anterior of the rabbit, the escape pathways of germs and their quantitative and qualitative distribution in eyes and blood has been studied in BCG-vaccinated and non-vaccinated animals while varying the amount of germs applied. As criteria, 35S concentration and the amount of 35S in ocular sections and in the blood which can be identified by means of distributing germs under the chosen conditions, have been detected. After only 10 minutes, elimination of germs is to be seen, continuing for a period longer than the 2 hours of post-injection period observed. Relatively high 35S concentrations indicate a long-term storage in the iris and in the ciliary body. The flow continues regularly but restrained via choroid and sclera into the blood. Flow velocity depends only within specific limits on the amount of germs injected into the anterior chamber. Under study conditions the flow mode via N.opticus and via lymphs is rather unimportant. The rest of the germs are distributed in the organism via blood vessels. A comparison of 35S concentration in sections of both eyes shows germ enrichment in tissues with sufficient blood supply, particularly in the choroid. Differences in germ distribution in vaccinated and non-vaccinated animals are not to be seen in the 35S distribution pattern. Neither higher nor lower germ doses indicate a stronger retention in the ocular sections of vaccinated animals. The necessity to complete this study by applying germ doses smaller than 1 mg (humidity weight) is stated pointing out technical difficulties involved while applying a test model. (orig.)

  10. The mineral nutrition of millet (Pennisetum-typhoides): Migration of 32P and 35S - Similarities with the migration of photosynthetic assimilates

    The paper describes a study of the transport of 32P and 35S injected into leaves or of 14CO2 absorbed thereby. It is shown that the radioisotopes mostly travel towards the upper parts of the plant and towards the seeds when they originate from upper leaves after male flowering. The later fillers, however, do not play any role as a reserve in the mineral or carbon nutrition of adult fillers. No appreciable absorption of 35S or 32P by the roots is observed after male flowering. (author)

  11. Contrast enhancement of intracranial lesions at 1.5 T: comparison among 2D spin echo, black-blood (BB) Cube, and BB Cube-FLAIR sequences

    Im, SungWoon; Ashikaga, Ryuichiro; Yagyu, Yukinobu; Hyodo, Tomoko; Imaoka, Izumi; Kumano, Seishi; Ishii, Kazunari; Murakami, Takamichi [Kinki University Faculty of Medicine, Department of Radiology, Osaka-Sayama, Osaka (Japan); Wakayama, Tetsuya; Miyoshi, Mitsuharu [GE Healthcare Japan, MR Applications and Workflow, Asia Pacific, Hino, Tokyo (Japan)

    2015-11-15

    The purpose of this study was to investigate the usefulness of T1W black-blood Cube (BB Cube) and T1W BB Cube fluid-attenuated inversion recovery (BB Cube-FLAIR) sequences for contrast-enhanced brain imaging, by evaluating flow-related artefacts, detectability, and contrast ratio (CR) of intracranial lesions among these sequences and T1W-SE. Phantom studies were performed to determine the optimal parameters of BB Cube and BB Cube-FLAIR. A clinical study in 23 patients with intracranial lesions was performed to evaluate the usefulness of these two sequences for the diagnosis of intracranial lesions compared with the conventional 2D T1W-SE sequence. The phantom study revealed that the optimal parameters for contrast-enhanced T1W imaging were TR/TE = 500 ms/minimum in BB Cube and TR/TE/TI = 600 ms/minimum/300 ms in BB Cube-FLAIR imaging. In the clinical study, the degree of flow-related artefacts was significantly lower in BB Cube and BB Cube-FLAIR than in T1W-SE. Regarding tumour detection, BB Cube showed the best detectability; however, there were no significant differences in CR among the sequences. At 1.5 T, contrast-enhanced BB Cube was a better imaging sequence for detecting brain lesions than T1W-SE or BB Cube-FLAIR. (orig.)

  12. Contrast enhancement of intracranial lesions at 1.5 T: comparison among 2D spin echo, black-blood (BB) Cube, and BB Cube-FLAIR sequences

    The purpose of this study was to investigate the usefulness of T1W black-blood Cube (BB Cube) and T1W BB Cube fluid-attenuated inversion recovery (BB Cube-FLAIR) sequences for contrast-enhanced brain imaging, by evaluating flow-related artefacts, detectability, and contrast ratio (CR) of intracranial lesions among these sequences and T1W-SE. Phantom studies were performed to determine the optimal parameters of BB Cube and BB Cube-FLAIR. A clinical study in 23 patients with intracranial lesions was performed to evaluate the usefulness of these two sequences for the diagnosis of intracranial lesions compared with the conventional 2D T1W-SE sequence. The phantom study revealed that the optimal parameters for contrast-enhanced T1W imaging were TR/TE = 500 ms/minimum in BB Cube and TR/TE/TI = 600 ms/minimum/300 ms in BB Cube-FLAIR imaging. In the clinical study, the degree of flow-related artefacts was significantly lower in BB Cube and BB Cube-FLAIR than in T1W-SE. Regarding tumour detection, BB Cube showed the best detectability; however, there were no significant differences in CR among the sequences. At 1.5 T, contrast-enhanced BB Cube was a better imaging sequence for detecting brain lesions than T1W-SE or BB Cube-FLAIR. (orig.)

  13. Nonrigid motion compensation in B-mode and contrast enhanced ultrasound image sequences of the carotid artery

    Carvalho, Diego D. B.; Akkus, Zeynettin; Bosch, Johan G.; van den Oord, Stijn C. H.; Niessen, Wiro J.; Klein, Stefan

    2014-03-01

    In this work, we investigate nonrigid motion compensation in simultaneously acquired (side-by-side) B-mode ultrasound (BMUS) and contrast enhanced ultrasound (CEUS) image sequences of the carotid artery. These images are acquired to study the presence of intraplaque neovascularization (IPN), which is a marker of plaque vulnerability. IPN quantification is visualized by performing the maximum intensity projection (MIP) on the CEUS image sequence over time. As carotid images contain considerable motion, accurate global nonrigid motion compensation (GNMC) is required prior to the MIP. Moreover, we demonstrate that an improved lumen and plaque differentiation can be obtained by averaging the motion compensated BMUS images over time. We propose to use a previously published 2D+t nonrigid registration method, which is based on minimization of pixel intensity variance over time, using a spatially and temporally smooth B-spline deformation model. The validation compares displacements of plaque points with manual trackings by 3 experts in 11 carotids. The average (+/- standard deviation) root mean square error (RMSE) was 99+/-74μm for longitudinal and 47+/-18μm for radial displacements. These results were comparable with the interobserver variability, and with results of a local rigid registration technique based on speckle tracking, which estimates motion in a single point, whereas our approach applies motion compensation to the entire image. In conclusion, we evaluated that the GNMC technique produces reliable results. Since this technique tracks global deformations, it can aid in the quantification of IPN and the delineation of lumen and plaque contours.

  14. Effects of cysteamine administration on the in vivo incorporation of (/sup 35/S)cysteine into somatostatin-14, somatostatin-28, arginine vasopressin, and oxytocin in rat hypothalamus

    Cameron, J.L.; Fernstrom, J.D.

    1986-09-01

    The effect of cysteamine injection on the in vivo incorporation of (/sup 35/S)cysteine into somatostatin-14 (SRIF-14), SRIF-28, arginine vasopressin (AVP), and oxytocin (OXT) in rat hypothalamus was studied. (/sup 35/S)Cysteine was injected into the third ventricle 1 h, 4 h, or 1 week after cysteamine (300 mg/kg, sc) injection; animals were killed 4 h later. The drug was found to substantially reduce immunoreactive SRIF levels, but not OXT or AVP, 4 h after its injection. Cysteamine also caused large reductions in label incorporation into SRIF-14, SRIF-28, and OXT 1 and 4 h after drug injection. However, (/sup 35/S)cysteine incorporation into AVP was increased substantially at these time points, while that into acid-precipitable protein was normal. One week after cysteamine injection, label incorporation into all hypothalamic peptides was normal. Cysteine specific activity was also measured after (/sup 35/S)cysteine injection and was found to be similar in treatment and control groups. The results suggest that cysteamine inhibits the syntheses of SRIF-14, SRIF-28, and OXT and stimulates that of AVP.

  15. Neutron-activated determination of chlorine, using the 35Cl(n,p)35S reaction as the basis, in thin coatings of silicon dioxide

    The neutron-activation determination of chlorine in thin coatings of silicon dioxide on silicon has been shown to be possible through the use of the 55Cl(n, P)35S reaction. The detection limit of chlorine is 3 x 10-9 g (5 x 1013 atoms)

  16. Uptake of [35S] carbonyl sulphide and [14C] carbon dioxide by crops in the vicinity of an advanced gas-cooled reactor

    The uptake into major crops of 35S and 14C released to the atmosphere during operation of Hinkley Point Nuclear Power Station was studied at an on-site experimental plot. Both radionuclides were measured in air and edible crop parts, while the effect of boiling on the 35S content of the latter was also investigated. The results were analysed in terms of air to crop transfer factors and the implications of these were assessed for both collective dose and dose to a hypothetical critical group. The transfer factor for 35S to green vegetables was found to be much smaller than believed previously, to the extent that the vegetable consumption pathway is of secondary importance to that of fresh milk. A possible reduction is indicated in critical group individual 35S doses of 20-60% and in collective dose by a factor of about 2, but vegetable consumption remains of potential radiological significance. It was confirmed that the specific activity approach used currently for assessments of 'first pass' dose from 14C releases is broadly correct, but a possible reduction of up to 3.5 times is indicated in the air-crop transfer for root vegetables. (author)

  17. Equilibration of [3H]glucosamine and [35S]sulfate with intracellular pools of UDP-N-acetylhexosamine and 3'-phosphoadenosine-5'-phosphosulfate (PAPS) in cultured fibroblasts

    Nucleotides and sugar nucleotides were extracted from cultures of human fibroblasts with perchloric acid, separated by isotachophoresis, and quantified by uv absorption analysis at 254 nm. ATP (936 pmol/micrograms DNA) was, as expected, the dominating nucleotide pool. The energy charge was estimated to 0.9. The UDP-N-acetylhexosamine pool was also a very prominent compound (596 pmol/micrograms DNA). After incubation of fibroblasts with [3H]glucosamine, more than 95% of the acid-soluble radioactivity was found in the UDP-N-acetylhexosamine pool. Incubation with [35S]sulfate resulted in the incorporation of [35S]sulfate into 3'-phosphoadenosine-5'-phosphosulfate (PAPS). The latter could, however, only be measured as radioactivity, as the amount was too small to be quantified as total mass. Pulse-labeling of fibroblasts with [35S]sulfate and [3H]glucosamine from 5 min to 16 h showed that [35S]PAPS was equilibrated in less than 10 min, while [3H]glucosamine required a longer time, 2-4 h, to attain a steady state with UDP-N-acetylhexosamine. [14C]Glucose required approximately the same time as [3H]glucosamine to reach steady state with UDP-acetylhexosamine, which suggests that the reason for the long equilibration time is the slow turnover of this pool

  18. Effects of cysteamine administration on the in vivo incorporation of [35S]cysteine into somatostatin-14, somatostatin-28, arginine vasopressin, and oxytocin in rat hypothalamus

    The effect of cysteamine injection on the in vivo incorporation of [35S]cysteine into somatostatin-14 (SRIF-14), SRIF-28, arginine vasopressin (AVP), and oxytocin (OXT) in rat hypothalamus was studied. [35S]Cysteine was injected into the third ventricle 1 h, 4 h, or 1 week after cysteamine (300 mg/kg, sc) injection; animals were killed 4 h later. The drug was found to substantially reduce immunoreactive SRIF levels, but not OXT or AVP, 4 h after its injection. Cysteamine also caused large reductions in label incorporation into SRIF-14, SRIF-28, and OXT 1 and 4 h after drug injection. However, [35S]cysteine incorporation into AVP was increased substantially at these time points, while that into acid-precipitable protein was normal. One week after cysteamine injection, label incorporation into all hypothalamic peptides was normal. Cysteine specific activity was also measured after [35S]cysteine injection and was found to be similar in treatment and control groups. The results suggest that cysteamine inhibits the syntheses of SRIF-14, SRIF-28, and OXT and stimulates that of AVP

  19. Three-dimensional gradient echo versus spin echo sequence in contrast-enhanced imaging of the pituitary gland at 3 T

    Kakite, Suguru, E-mail: sugkaki@med.tottori-u.ac.jp [Division of Radiology, Department of Pathophysiological and Therapeutic Science, Faculty of Medicine, Tottori University, 36-1, Nishicho, Yonago 683-8503 (Japan); Fujii, Shinya [Division of Radiology, Department of Pathophysiological and Therapeutic Science, Faculty of Medicine, Tottori University, 36-1, Nishicho, Yonago 683-8503 (Japan); Kurosaki, Masamichi [Department of Neurosurgery, Faculty of Medicine, Tottori University, 36-1, Nishicho, Yonago 683-8503 (Japan); Kanasaki, Yoshiko; Matsusue, Eiji; Kaminou, Toshio; Ogawa, Toshihide [Division of Radiology, Department of Pathophysiological and Therapeutic Science, Faculty of Medicine, Tottori University, 36-1, Nishicho, Yonago 683-8503 (Japan)

    2011-07-15

    Introduction: To clarify whether a three-dimensional-gradient echo (3D-GRE) or spin echo (SE) sequence is more useful for evaluating sellar lesions on contrast-enhanced T1-weighted MR imaging at 3.0 Tesla (T). Methods: We retrospectively assessed contrast-enhanced T1-weighted images using 3D-GRE and SE sequences at 3.0 T obtained from 33 consecutive patients with clinically suspected sellar lesions. Two experienced neuroradiologists evaluated the images qualitatively in terms of the following criteria: boundary edge of the cavernous sinus and pituitary gland, border of sellar lesions, delineation of the optic nerve and cranial nerves within the cavernous sinus, susceptibility and flow artifacts, and overall image quality. Results: At 3.0 T, 3D-GRE provided significantly better images than the SE sequence in terms of the border of sellar lesions, delineation of cranial nerves, and overall image quality; there was no significant difference regarding the boundary edge of the cavernous sinus and pituitary gland. In addition, the 3D-GRE sequence showed fewer pulsation artifacts but more susceptibility artifacts. Conclusion: Our results indicate that 3D-GRE is the more suitable sequence for evaluating sellar lesions on contrast-enhanced T1-weighted imaging at 3.0 T.

  20. Automatic classification of lung tumour heterogeneity according to a visual-based score system in dynamic contrast enhanced CT sequences

    Bevilacqua, Alessandro; Baiocco, Serena

    2016-03-01

    Computed tomography (CT) technologies have been considered for a long time as one of the most effective medical imaging tools for morphological analysis of body parts. Contrast Enhanced CT (CE-CT) also allows emphasising details of tissue structures whose heterogeneity, inspected through visual analysis, conveys crucial information regarding diagnosis and prognosis in several clinical pathologies. Recently, Dynamic CE-CT (DCE-CT) has emerged as a promising technique to perform also functional hemodynamic studies, with wide applications in the oncologic field. DCE-CT is based on repeated scans over time performed after intravenous administration of contrast agent, in order to study the temporal evolution of the tracer in 3D tumour tissue. DCE-CT pushes towards an intensive use of computers to provide automatically quantitative information to be used directly in clinical practice. This requires that visual analysis, representing the gold-standard for CT image interpretation, gains objectivity. This work presents the first automatic approach to quantify and classify the lung tumour heterogeneities based on DCE-CT image sequences, so as it is performed through visual analysis by experts. The approach developed relies on the spatio-temporal indices we devised, which also allow exploiting temporal data that enrich the knowledge of the tissue heterogeneity by providing information regarding the lesion status.

  1. Proteome scale census of major facilitator superfamily transporters in Trichoderma reesei using protein sequence and structure based classification enhanced ranking.

    Chaudhary, Nitika; Kumari, Indu; Sandhu, Padmani; Ahmed, Mushtaq; Akhter, Yusuf

    2016-07-01

    Trichoderma spp. have been acknowledged as potent bio-control agents against microbial pathogens and also as plant growth promoters. Various secondary metabolites are attributed for these beneficial activities. Major facilitator superfamily (MFS) includes the large proportion of efflux-pumps which are linked with membrane transport of these secondary metabolites. We have carried out a proteome-wide identification of MFS transporters using protein sequence and structure based hierarchical method in Trichoderma reesei. 448 proteins out of 9115 were detected to carry transmembrane helices. MFS specific intragenic gene duplication and its context with transport function have been presented. Finally, using homology based techniques, domains and motifs of MFS families have been identified and utilized to classify them. From query dataset of 448 transmembrane proteins, 148 proteins are identified as potential MFS transporters. Sugar porter, drug: H(+) antiporter-1, monocarboxylate porter and anion: cation symporter emerged as major MFS families with 51, 35, 17 and 11 members respectively. Representative protein tertiary structures of these families are homology modeled for structure-function analysis. This study may help to understand the molecular basis of secretion and transport of agriculturally valuable secondary metabolites produced by these bio-control fungal agents which may be exploited in future for enhancing its biotechnological applications in eco-friendly sustainable development. PMID:27041239

  2. Molecular cloning and characterization of a novel sequence, vof-16, with enhanced expression in permanent ischemic rat brain.

    Tohda, Michihisa; Watanabe, Hiroshi

    2004-08-01

    We reported previously that chronic hypoperfusion induced by permanent occlusion of the bilateral common carotid arteries (2VO) in rats caused progressive cognitive deficits and neuronal damage in the hippocampus and the white matter. These changes are similar to those observed in human dementia. Reverse transcription-polymerase chain reaction (RT-PCR) differential display was carried out to identify mRNAs encoding the intrinsic factors involved in permanent ischemia from the 2VO rat brain. Over 20 clones which showed different expression levels in 2VO and sham-operated rats were isolated. One of these, named vof-16, was markedly enhanced the expression by 2VO. The whole sequence of vof-16 mRNA was 2098 nt. The distribution of vof-16 transcripts was examined by RT-PCR and in situ hybridization. The results revealed that vof-16 was abundant in the hippocampus, the tenia tecta, the piriform cortex and the area around the aorta. The expression levels of vof-16 in 2VO and sham-operated rat hippocampus were determined by a quantitative PCR method. The expression was abundant in the hippocampus of rats with cognitive impairment induced by 2VO. In contrast, the expression levels of vof-16 were lower in the 2VO rats with no impairment and in sham-operated rats. These results suggest that the expression levels of vof-16 may be related to the cognitive impairment induced by chronic ischemia after 2VO. PMID:15305027

  3. Technical innovation in dynamic contrast-enhanced magnetic resonance imaging of musculoskeletal tumors: an MR angiographic sequence using a sparse k-space sampling strategy

    We demonstrate the clinical use of an MR angiography sequence performed with sparse k-space sampling (MRA), as a method for dynamic contrast-enhanced (DCE)-MRI, and apply it to the assessment of sarcomas for treatment response. Three subjects with sarcomas (2 with osteosarcoma, 1 with high-grade soft tissue sarcomas) underwent MRI after neoadjuvant therapy/prior to surgery, with conventional MRI (T1-weighted, fluid-sensitive, static post-contrast T1-weighted sequences) and DCE-MRI (MRA, time resolution = 7-10 s, TR/TE 2.4/0.9 ms, FOV 40 cm2). Images were reviewed by two observers in consensus who recorded image quality (1 = diagnostic, no significant artifacts, 2 = diagnostic, 75 % with good response, >75 % with poor response). DCE-MRI findings were concordant with histological response (arterial enhancement with poor response, no arterial enhancement with good response). Unlike conventional DCE-MRI sequences, an MRA sequence with sparse k-space sampling is easily integrated into a routine musculoskeletal tumor MRI protocol, with high diagnostic quality. In this preliminary work, tumor enhancement characteristics by DCE-MRI were used to assess treatment response. (orig.)

  4. Estimation of perilymph enhancement after intratympanic administration of Gd-DTPA by fast T1-mapping with a dual flip angle 3D spoiled gradient echo sequence

    Eleven patients with suspected Meniere's disease received intratympanic (IT) administration of gadolinium (gadopentetate dimeglumine; Gd) prior to acquisition of 3-dimensional (3D) fluid-attenuated inversion recovery (FLAIR), 3D real inversion recovery (IR), and fast T1 mapping by dual flip angle 3D gradient echo (FT1-map) imaging sequences to evaluate the degree of perilymph enhancement. Though 3-dimensional FLAIR could detect lower concentrations of gadolinium than 3D real IR, in 2 patients, poor enhancement still prevented visualization of the endolymphatic space using 3D FLAIR. We could predict poor contrast enhancement in these 2 patients using the FT1-map technique. (author)

  5. Differentiation of activities within the GABAA-chloride ionophore complex by means of 35-S-TBPS binding.

    Lloyd, K G; Danielou, G; Thuret, F

    1988-01-01

    From the above results, it is evident that both alpidem and zolpidem modulate the GABAA receptor linked chloride ionophore in an allosteric manner via omega 1 anxiolytic/hypnotic recognition sites. As both are highly specific for the omega 1 site, with little affinity for the omega 2 site, it appears that omega 1 site activation is sufficient to fully engage the various linkages within the GABAA receptor supramolecular complex, resulting in modulation of the chloride ionophore. This action is related to an enhanced affinity of the recognition site for TBPS. Under the present conditions (high sodium chloride, frozen well-washed membranes), several (but not all, e.g. zolpidem) anxiolytics and hypnotics decreased TBPS binding at very high (100-500 microM) concentrations. This effect is unlikely related to the pharmacological activity of these compounds, as it is insensitive to flumazenil and occurs only at concentrations which would be supra-toxic. In contrast, the enhancement of TBPS binding by these anxiolytics and hypnotics occurs within the range of, and correlates with, their therapeutic plasma levels and their affinity for omega 1/omega 2 receptors. The present findings suggest that a different degree of linkage for different compounds occurs between the GABAA receptor and the omega 1/omega 2 receptor mediated enhancement of TBPS binding, as the action of alpidem is completely reversed by bicuculline, whereas for zopidem and flunitrazepam a component of the TBPS enhancement is bicuculline insensitive. A Ro 5-4864 sensitive site (probably not the omega 3 site) occurs with the GABAA receptor supramolecular complex, which apparently participates in the enhancement of TBPS binding.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2459920

  6. Characterization of adenomas and metastases in the adrenals: comparison between spin-echo sequences and Gd-DTPA enhanced dynamic studies

    Nineteen patients with adrenal masses (9 metastases and 10 adenomas) were studied with magnetic resonance spin-echo sequences and with dynamic perfusion with gradient-echo (GE) sequences after gadolinium diethylenetriamine-pentaacetic acid injection. In studies following Gd-DTPA injection, sequential imaging during suspended respiration was obtained. Signal intensities of adrenal masses and retroperitonel fat were measured in the region of interest. The relative signal intensities of lesion and fat were determined and compared in dynamic contrast-enhanced studies. Results were tabulated and plotted in diagrams as a function of time. Sixteen lesions out of 19 were correctly classified by means of SE sequences, with an 84% diagnostic accuracy, whereas 18 out of 19 lesions were characterized by means of GE sequences, with a 95% accuracy. In the dynamic study, adenomas showed mild enhancement and quicker washout than metastases which, in turn, showed a strong enhancement and a much slower washout. The most significant values were obtained in scans taken after 10 min, where adenomas showed signal to lesion/signal to fat values lower than 1.1, whereas metastases showed much higher values in all cases. (orig.)

  7. In planta analysis of a cis-regulatory cytokinin response motif in Arabidopsis and identification of a novel enhancer sequence.

    Ramireddy, Eswarayya; Brenner, Wolfram G; Pfeifer, Andreas; Heyl, Alexander; Schmülling, Thomas

    2013-07-01

    The phytohormone cytokinin plays a key role in regulating plant growth and development, and is involved in numerous physiological responses to environmental changes. The type-B response regulators, which regulate the transcription of cytokinin response genes, are a part of the cytokinin signaling system. Arabidopsis thaliana encodes 11 type-B response regulators (type-B ARRs), and some of them were shown to bind in vitro to the core cytokinin response motif (CRM) 5'-(A/G)GAT(T/C)-3' or, in the case of ARR1, to an extended motif (ECRM), 5'-AAGAT(T/C)TT-3'. Here we obtained in planta proof for the functionality of the latter motif. Promoter deletion analysis of the primary cytokinin response gene ARR6 showed that a combination of two extended motifs within the promoter is required to mediate the full transcriptional activation by ARR1 and other type-B ARRs. CRMs were found to be over-represented in the vicinity of ECRMs in the promoters of cytokinin-regulated genes, suggesting their functional relevance. Moreover, an evolutionarily conserved 27 bp long T-rich region between -220 and -193 bp was identified and shown to be required for the full activation by type-B ARRs and the response to cytokinin. This novel enhancer is not bound by the DNA-binding domain of ARR1, indicating that additional proteins might be involved in mediating the transcriptional cytokinin response. Furthermore, genome-wide expression profiling identified genes, among them ARR16, whose induction by cytokinin depends on both ARR1 and other specific type-B ARRs. This together with the ECRM/CRM sequence clustering indicates cooperative action of different type-B ARRs for the activation of particular target genes. PMID:23620480

  8. Distributions of 35S-sulfate and 3H-glucosamine in the angular region of the hamster: light and electron microscopic autoradiography

    The distribution of 35S-sulfate and 3H-glucosamine in the angular region of the hamster was studied by light and electron microscopic autoradiography following intraperitoneal injection of these compounds to hamsters. Exposed silver grains of 35S-sulfate were concentrated in the trabecular meshwork, sclera, and cornea, and grains of 3H-glucosamine were localized in the trabecular region. The radioactivity of both isotopes was observed in the Golgi apparatuses of the endothelial cells of the angular aqueous plexus and the trabecular meshwork. The grains were noted over the entire cytoplasm, except for the nucleus, and then were incorporated into the amorphous substance and collagen fibers in the region adjacent to the angular aqueous sinus. These results suggest that endothelial cells in the angular region synthesize and secrete the sulfated glycosaminoglycans and hyaluronic acid

  9. Cocaine alters mu but not delta or kappa opioid receptor-stimulated in situ [35S]GTPgammaS binding in rat brain.

    Schroeder, Joseph A; Niculescu, Michelle; Unterwald, Ellen M

    2003-01-01

    Chronic cocaine administration produces alterations in mu and kappa opioid receptor density as well as striatal and accumbens opioid-regulated adenylyl cyclase activity, suggesting a psychostimulant responsive interaction between opioidergic and dopaminergic systems. Stimulation of G-protein-coupled opioid receptors inhibits adenylyl cyclase production of cyclic AMP. The present study employed in situ [(35)S]GTPgammaS binding to measure opioid receptor-stimulated activation of G-proteins in response to acute and chronic cocaine exposure. Male Fischer rats received acute (1 or 3 days) or chronic (14 days) binge pattern cocaine administration. Three and 14 days of cocaine injections resulted in greater increases in the ability of the mu receptor agonist DAMGO to stimulate [(35)S]GTPgammaS binding in both the core and the shell of the nucleus accumbens, all regions of the caudate putamen and the cingulate cortex compared with saline-matched controls. The greatest increases in DAMGO-stimulated [(35)S]GTPgammaS binding were observed in the dorsal areas of the caudate putamen in animals that received 14 days of cocaine. No significant changes in delta (DPDPE), or kappa (dynorphin A(1-17)) receptor-stimulated [(35)S]GTPgammaS binding were found in any brain region in response to cocaine administration. These results demonstrate that binge pattern cocaine administration induce changes in mu but not delta or kappa opioid receptor-mediated G-protein activity. This study provides support for the hypothesis that the addictive properties of both psychostimulants and opiates may share common neurochemical signaling substrates. PMID:12422370

  10. Ovarian steroid effects on gonadotrophin output, and on the incorporation of 35S from methionine into protein of discrete brain areas and the anterior pituitary

    The effects of various ovarian hormones administered on the morning of pro-oestrus on gonadotrophin levels and the incorporation of 35S from methionine into protein of discrete areas of the brain and the anterior pituitary were investigated at 15.00 h of the same day in femal rats. The hormones which were investigated in this study could be divided in general into two groups according to their actions. The first group, consisting of oestradiol-17β and progesterone, tended to advance the preovulatory surge of luteinizing hormone (LH) by 3-6 h from 18.00-21.00 h. together with the peaks of [35S]incorporation in the median eminence area and the anterior pituitary which normally accompany the LH surge. The second group, consisting of the LH-stimulated reduced progesterone metabolites, 5α-pregnane-3,20-dione (pregnanedione) and 20α-hydroxy-pregn-4-en-3-one (dihydroprogesterone), tended to inhibit serum gonadotrophin levels as well as inhibiting the pro-oestrous increase of [35S]incorporation in the median eminence area and in the amygdala, but not in the preoptic area and the anterior pituitary. On the afternoon of pro-oestrus in intact animals, luteinizing hormonereleasing hormone or LH administration had the same effect on [35S]incorporation in the brain as did the progesterone metabolites, though this effect was not observed it the animals had been ovariectomized a few hours beforehand. It is suggested that certain ovarian hormones are involved in the neural events which induce the pre-ovulatory LH surge, while others are associated with neural events which terminate the stimulus for the LH surge. (author)

  11. Elucidation of coal liquefaction mechanism by the use of tritium and 35S tracer methods. Effects of pyrrhotite and sulfur on hydrogen transfer in coal liquefaction

    Effects of addition of the catalyst (pyrrhotite) and sulfur on hydrogen transfer in liquefaction of Taiheiyo coal were investigated using tritium and 35S. The coal liquefaction was performed at the initial pressure of 5.9 MPa and at 400degC for 30 min with tetralin solvent and tritium-labelled hydrogen, with or without the synthesized pyrrhotite catalyst and sulfur (or 35S-labelled sulfur). The specific activities of tritium and 35S in the reaction products were measured with a liquid scintillation counter. Amounts of exchanged and transferred hydrogens between the gas phase and coal/solvent, were calculated from the distributions of tritium and changes in the composition of products. In the reaction with tritiated hydrogen and solvent, the dehydrogenation of tetralin to produce naphthalene and the hydrogen exchange reaction between gas phase and solvent were promoted by added catalyst and sulfur. Added sulfur produced hydrogen sulfide mainly with hydrogen of solvent. A part of added sulfur participated in the sulfur exchange reaction with the pyrrhotite catalyst. In the reaction with tritiated hydrogen, solvent and coal, the hydrogen addition and exchange to coal and liquefaction products increased with the addition of catalyst and sulfur. It was suggested the sulfur promoted the formation of tetralyl radical in the hydrogen transfer from solvent to coal. (author)

  12. An estimation of the radioactive 35S emitted into the atmospheric from the Fukushima Daiichi Nuclear Power Plant by using a numerical simulation global transport

    We present a numerical study carried out with the SPRINTARS model modified to account for the radioactive decay of 35S compounds emitted from the Fukushima Daiichi nuclear power plant station after the hydrogen and vapor blast. The transport dynamics of the released material reproduced previous field observations. Four different emission scenarios were compared to the measurements of atmospheric 35S in sulfate collected in La Jolla, Tsukuba, Kashiwa and Yokohama. Linear regressions of the relation between emitted and transported material that reached the sampling sites were used to estimate the amount of 35S atoms and the amount of neutrons released in to the atmosphere. We estimate that a lower limit of 1.9 × 101635S atoms sec-1 were released after the events in March and this flux dropped to 4 - 39 × 101435S atoms sec-1 at the end of the month. Based on this calculations we estimated a lower limit of 5.2 × 1021 slow neutrons m-2 sec-1 were emitted from the nuclear fuel rods to the sea water injected in the reactors after the events in March. (author)

  13. Quantitative assessment of hepatic function: modified look-locker inversion recovery (MOLLI) sequence for T1 mapping on Gd-EOB-DTPA-enhanced liver MR imaging

    Yoon, Jeong Hee [Seoul National University Hospital, Department of Radiology, Seoul (Korea, Republic of); Lee, Jeong Min; Han, Joon Koo; Choi, Byung Ihn [Seoul National University Hospital, Department of Radiology, Seoul (Korea, Republic of); Seoul National University College of Medicine, Institute of Radiation Medicine, Jongno-gu, Seoul (Korea, Republic of); Paek, Munyoung [Siemens Healthcare, Seoul (Korea, Republic of)

    2016-06-15

    To determine whether multislice T1 mapping of the liver using a modified look-locker inversion recovery (MOLLI) sequence on gadoxetic acid-enhanced magnetic resonance imaging (MRI) can be used as a quantitative tool to estimate liver function and predict the presence of oesophageal or gastric varices. Phantoms filled with gadoxetic acid were scanned three times using MOLLI sequence to test repeatability. Patients with chronic liver disease or liver cirrhosis who underwent gadoxetic acid-enhanced liver MRI including MOLLI sequence at 3 T were included (n = 343). Pre- and postcontrast T1 relaxation times of the liver (T1liver), changes between pre- and postcontrast T1liver (ΔT1liver), and adjusted postcontrast T1liver (postcontrast T1liver-T1spleen/T1spleen) were compared among Child-Pugh classes. In 62 patients who underwent endoscopy, all T1 parameters and spleen sizes were correlated with varices. Phantom study showed excellent repeatability of MOLLI sequence. As Child-Pugh scores increased, pre- and postcontrast T1liver were significantly prolonged (P < 0.001), and ΔT1liver and adjusted postcontrast T1liver decreased (P< 0.001). Adjusted postcontrast T1liver and spleen size were independently associated with varices (R{sup 2} = 0.29, P < 0.001). T1 mapping of the liver using MOLLI sequence on gadoxetic acid-enhanced MRI demonstrated potential in quantitatively estimating liver function, and adjusted postcontrast T1liver was significantly associated with varices. (orig.)

  14. Analysis of DNA sequences which regulate the transcription of herpes simplex virus immediate early gene 3: DNA sequences required for enhancer-like activity and response to trans-activation by a virion polypeptide.

    Bzik, D J; Preston, C M

    1986-01-24

    The far upstream region of herpes simplex virus (HSV) immediate early (IE) gene 3 has previously been shown to increase gene expression in an enhancer-like manner, and to contain sequences which respond to stimulation of transcription by a virion polypeptide, Vmw65. To analyse the specific DNA sequences which mediate these functions, sequential deletions from each end of the far upstream region were made. The effects of the deletions on transcription in the absence or presence of the Vmw65 were measured by use of a transient expression assay. The enhancer-like activity was due to three separable elements, whereas two additional DNA regions were involved in the response to Vmw65. One of the responding elements corresponded to an AT-rich consensus (TAATGARATTC, where R = purine) present in all IE gene far upstream regions, and the other was a GA-rich sequence also present in IE genes 2 and 4/5. The TAATGARATTC element could mediate responsiveness to Vmw65 but it was fully active only in the presence of the GA-rich element. The GA-rich element was unable to confer a strong response alone but could activate an otherwise nonfunctional homologue of TAATGARATTC. PMID:3003700

  15. Modulations of RNA sequences by cytokinin in pumpkin cotyledons

    Polyadenylated mRNAs from excised pumpkin cotyledons treated with or without 10-4 M benzyladenine (BA) for various time periods in suspension culture were assayed by in vitro translation in the presence of [35S] methionine. The radioactive polypeptides were analyzed by one- and two-dimensional polyacrylamide gel electrophoresis. Specific sequences of mRNAs were enhanced, reduced, induced, or suppressed by the hormone within 60 min of the application of BA to the cotyledons. Four independent cDNA clones of cytokinin-modulated mRNAs have been selected and characterized. RNA blot hybridization using the four cDNA probes also indicates that the levels of specific mRNAs are modulated upward or downward by the hormone

  16. Methylation-sensitive linking libraries enhance gene-enriched sequencing of complex genomes and map DNA methylation domains

    Bharti Arvind K

    2008-12-01

    Full Text Available Abstract Background Many plant genomes are resistant to whole-genome assembly due to an abundance of repetitive sequence, leading to the development of gene-rich sequencing techniques. Two such techniques are hypomethylated partial restriction (HMPR and methylation spanning linker libraries (MSLL. These libraries differ from other gene-rich datasets in having larger insert sizes, and the MSLL clones are designed to provide reads localized to "epigenetic boundaries" where methylation begins or ends. Results A large-scale study in maize generated 40,299 HMPR sequences and 80,723 MSLL sequences, including MSLL clones exceeding 100 kb. The paired end reads of MSLL and HMPR clones were shown to be effective in linking existing gene-rich sequences into scaffolds. In addition, it was shown that the MSLL clones can be used for anchoring these scaffolds to a BAC-based physical map. The MSLL end reads effectively identified epigenetic boundaries, as indicated by their preferential alignment to regions upstream and downstream from annotated genes. The ability to precisely map long stretches of fully methylated DNA sequence is a unique outcome of MSLL analysis, and was also shown to provide evidence for errors in gene identification. MSLL clones were observed to be significantly more repeat-rich in their interiors than in their end reads, confirming the correlation between methylation and retroelement content. Both MSLL and HMPR reads were found to be substantially gene-enriched, with the SalI MSLL libraries being the most highly enriched (31% align to an EST contig, while the HMPR clones exhibited exceptional depletion of repetitive DNA (to ~11%. These two techniques were compared with other gene-enrichment methods, and shown to be complementary. Conclusion MSLL technology provides an unparalleled approach for mapping the epigenetic status of repetitive blocks and for identifying sequences mis-identified as genes. Although the types and natures of

  17. 3D Whole-Heart Coronary MR Angiography at 1.5T in Healthy Volunteers: Comparison between Unenhanced SSFP and Gd-Enhanced FLASH Sequences

    To validate the optimal cardiac phase and appropriate acquisition window for three-dimensional (3D) whole-heart coronary magnetic resonance angiography (MRA) with a steady-state free precession (SSFP) sequence, and to compare image quality between SSFP and Gd-enhanced fast low-angle shot (FLASH) MR techniques at 1.5 Tesla (T). Thirty healthy volunteers (M:F 25:5; mean age, 35 years; range, 24-54 years) underwent a coronary MRA at 1.5T. 3D whole-heart coronary MRA with an SSFP was performed at three different times: 1) at end-systole with a narrow (120-msec) acquisition window (ESN), 2) mid-diastole with narrow acquisition (MDN); and 3) mid-diastole with wide (170-msec) acquisition (MDW). All volunteers underwent a contrast enhanced coronary MRA after undergoing an unenhanced 3D true fast imaging with steady-state precession (FISP) MRA three times. A contrast enhanced coronary MRA with FLASH was performed during MDN. Visibility of the coronary artery and image quality were evaluated for 11 segments, as suggested by the American Heart Association. Image quality was scored by a five-point scale (1 = not visible to 5 = excellent). The signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) were evaluated at the proximal coronary arteries. The SSFP sequence rendered higher visibility coronary segments, higher image quality, as well as higher SNR and CNR than the Gd-enhanced FLASH technique at 1.5T (p < 0.05). The visibility of coronary segments, image quality, SNR and CNR in the ESN, MDN and MDW with SSFP sequence did not differ significantly. An SSFP sequence provides an excellent method for the 3D whole-heart coronary MRA at 1.5T. Contrast enhanced coronary MRA using the FLASH sequence does not help improve the visibility of coronary segments, image quality, SNR or CNR on the 3D whole-heart coronary MRA.

  18. Evaluation of Dixon Sequence on Hybrid PET/MR Compared with Contrast-Enhanced PET/CT for PET-Positive Lesions

    Jeong, Ju Hye; Cho, Ihn Ho; Kong, Eun Jung; Chun, Kyung Ah [Yeungnam Univ. Hospital, Daegu (Korea, Republic of)

    2014-03-15

    Hybrid positron emission tomography and magnetic resonance (PET/MR) imaging performs a two-point Dixon MR sequence for attenuation correction. However, MR data in hybrid PET/MR should provide anatomic and morphologic information as well as an attenuation map. We evaluated the Dixon sequence of hybrid PET/MR for anatomic correlation of PET-positive lesions compared with contrast-enhanced PET/computed tomography (CT) in patients with oncologic diseases. Twelve patients underwent a single injection, dual imaging protocol. PET/CT was performed with an intravenous contrast agent (85±13 min after {sup 18}F-FDG injection of 403± 45 MBq) and then (125±19 min after injection) PET/MR was performed. Attenuation correction and anatomic allocation of PET were performed using contrast-enhanced CT for PET/CT and Dixon MR sequence for hybrid PET/MR. The Dixon MR sequence and contrast-enhanced CT were compared for anatomic correlation of PET-positive lesions (scoring scale ranging from 0 to 3 for visual ratings). Additionally, standardized uptake values (SUVs) for the detected lesions were assessed for quantitative comparison. Both hybrid PET/MR and contrast-enhanced PET/CT identified 55 lesions with increased FDG uptake in ten patients. In total, 28 lymph nodes, 11 bone lesions, 3 dermal nodules, 3 pleural thickening lesions, 2 thyroid nodules, 1 pancreas, 1 liver, 1 ovary, 1 uterus, 1 breast, 1 soft tissue and 2 lung lesions were present. The best performance was observed for anatomic correlation of PET findings by the contrast-enhanced CT scans (contrast-enhanced CT, 2.64± 0.70; in-phase, 1.29±1.01; opposed-phase, 1.29±1.15; water-weighted, 1.71±1.07; fat weighted, 0.56±1.03). A significant difference was observed between the scores obtained from the contrast-enhanced CT and all four coregistered Dixon MR images. Quantitative evaluation revealed a high correlation between the SUVs measured with hybrid PET/MR (SUVmean, 2.63±1.62; SUVmax, 4.30±2.88) and contrast-enhanced

  19. Comparison of different magnetic resonance cholangiography techniques in living liver donors including Gd-EOB-DTPA enhanced T1-weighted sequences.

    Sonja Kinner

    Full Text Available Preoperative evaluation of potential living liver donors (PLLDs includes the assessment of the biliary anatomy to avoid postoperative complications. Aim of this study was to compare T2-weighted (T2w and Gd-EOB-DTPA enhanced T1-weighted (T1w magnetic resonance cholangiography (MRC techniques in the evaluation of PLLDs.30 PLLDs underwent MRC on a 1.5 T Magnetom Avanto (Siemens, Erlangen, Germany using (A 2D T2w HASTE (Half Fourier Acquisition Single Shot Turbo Spin Echo fat saturated (fs in axial plane, (B 2D T2w HASTE fs thick slices in coronal plane, (C free breathing 3D T2w TSE (turbo spin echo RESTORE (high-resolution navigator corrected plus (D maximum intensity projections (MIPs, (E T2w SPACE (sampling perfection with application optimized contrasts using different flip angle evolutions plus (F MIPs and (G T2w TSE BLADE as well as Gd-EOB-DTPA T1w images without (G and with (H inversion recovery. Contrast enhanced CT cholangiography served as reference imaging modality. Two independent reviewers evaluated the biliary tract anatomy on a 5-point scale subjectively and objectively. Data sets were compared using a Mann-Whitney-U-test. Kappa values were also calculated.Source images and maximum intensity projections of 3D T2w TSE sequences (RESTORE and SPACE proved to be best for subjective and objective evaluation directly followed by 2D HASTE sequences. Interobserver variabilities were good to excellent (k = 0.622-0.804.3D T2w sequences are essential for preoperative biliary tract evaluation in potential living liver donors. Furthermore, our results underline the value of different MRCP sequence types for the evaluation of the biliary anatomy in PLLDs including Gd-EOB-DTPA enhanced T1w MRC.

  20. Cerebral staging of lung cancer: is one single contrast-enhanced T1-weighted three-dimensional gradient-echo sequence sufficient?

    Gadolinium-enhanced magnetic resonance imaging (MRI) is the gold standard for cerebral staging in thoracic oncology. We hypothesize that a minimalist examination, consisting of a single contrast-enhanced T1-weighted three-dimensional gradient-echo sequence (CE 3D-GRE), would be sufficient for the cerebral staging of nonsymptomatic lung cancer patients. Seventy nonsymptomatic patients (50 % men; 62 years ± 10.2) referred for cerebral staging of a lung cancer were retrospectively included. All underwent a standard 3 T MRI examination with T1, FLAIR, T2* GRE, diffusion, and CE 3D-GRE sequences, for a total examination time of 20 min. The sole CE 3D-GRE (acquisition time: 6 min) was extracted and blindly interpreted by two radiologists in search of brain metastases. Hemorrhagic features of potential lesions and relevant incidental findings were also noted. Discrepant cases were reviewed by a third reader. The full MRI examination and follow-up studies were used as a reference to calculate sensitivity and specificity of the sole CE 3D-GRE. Thirty-eight point six percent (27 out of 70) of the patients had brain metastases. Performances and reader's agreement with the sole CE 3D-GRE sequence were excellent for the diagnosis of brain metastases (sensitivity = 96.3 %, specificity = 100 %, κ = 0.91) and incidental findings (sensitivity = 85.7 %, specificity = 100 %, κ = 0.62) but insufficient for the identification of hemorrhages within the metastases (sensitivity = 33.3 %, specificity = 85.7 %, κ = 0.47). In the specific case of lung cancer, cerebral staging in nonsymptomatic patients can be efficiently achieved with a minimalistic protocol consisting of a single CE 3D-GRE sequence, completed if positive with a T2* sequence for hemorrhagic assessment, thus halving appointment delays. (orig.)

  1. Cerebral staging of lung cancer: is one single contrast-enhanced T1-weighted three-dimensional gradient-echo sequence sufficient?

    Ohana, Mickael; Jeung, Mi-Young; Roy, Catherine [Nouvel Hopital Civil-Hopitaux Universitaires de Strasbourg, Service de Radiologie B/Radiology Department, Strasbourg (France); Bazille, Gauthier [Clinique Saint Anne-Groupe Radiologique MIM, Strasbourg (France)

    2014-08-15

    Gadolinium-enhanced magnetic resonance imaging (MRI) is the gold standard for cerebral staging in thoracic oncology. We hypothesize that a minimalist examination, consisting of a single contrast-enhanced T1-weighted three-dimensional gradient-echo sequence (CE 3D-GRE), would be sufficient for the cerebral staging of nonsymptomatic lung cancer patients. Seventy nonsymptomatic patients (50 % men; 62 years ± 10.2) referred for cerebral staging of a lung cancer were retrospectively included. All underwent a standard 3 T MRI examination with T1, FLAIR, T2* GRE, diffusion, and CE 3D-GRE sequences, for a total examination time of 20 min. The sole CE 3D-GRE (acquisition time: 6 min) was extracted and blindly interpreted by two radiologists in search of brain metastases. Hemorrhagic features of potential lesions and relevant incidental findings were also noted. Discrepant cases were reviewed by a third reader. The full MRI examination and follow-up studies were used as a reference to calculate sensitivity and specificity of the sole CE 3D-GRE. Thirty-eight point six percent (27 out of 70) of the patients had brain metastases. Performances and reader's agreement with the sole CE 3D-GRE sequence were excellent for the diagnosis of brain metastases (sensitivity = 96.3 %, specificity = 100 %, κ = 0.91) and incidental findings (sensitivity = 85.7 %, specificity = 100 %, κ = 0.62) but insufficient for the identification of hemorrhages within the metastases (sensitivity = 33.3 %, specificity = 85.7 %, κ = 0.47). In the specific case of lung cancer, cerebral staging in nonsymptomatic patients can be efficiently achieved with a minimalistic protocol consisting of a single CE 3D-GRE sequence, completed if positive with a T2* sequence for hemorrhagic assessment, thus halving appointment delays. (orig.)

  2. Quantitation of sodium 35S-sulphate in nano mole levels by spectrophotometric assay using barium chloranilate and liquid scintillation counting

    In this assay, the absorbance of chloranilic acid released from buffered suspension of barium chloranilate, taken in excess, on reaction with solutions of known concentrations of authentic sodium sulphate (2-10 fg; 14-70 nanomoles) and that of sodium 35S-sulphate of known radioactivity (10-20 mci; 370-740 mBq) is measured spectrophotometrically and the quantitation done accordingly. The amount of chloranilic acid released has been found to be directly proportional to the amount of sulphate present in the original sample solution. (author). 8 refs., 1 tab

  3. A loss-free measurement of the beta-spectrum of 35S and the origin of the 17 keV neutrino signals

    We present measurements of the β-spectrum of 35S, using two energy sensitive silicon detectors in a 4π configuration. A 7 Tesla magnetic field guides the electrons to the detectors. Electrons which are backscattered in one detector are counted in the other detector. No baffles or collimators are needed. In the sum spectra all effects due to electron backscattering are eliminated. No evidence for heavy neutrinos is observed. When we remove the backscattering suppression, the data show a deviation from the usual electron spectrum that could be misinterpreted as an admixture of a heavy neutrino. (authors). 11 refs., 5 figs

  4. Differential regulation of serotonin-1A receptor stimulated [35S]GTPγS binding in the dorsal raphe nucleus by citalopram and escitalopram

    Rossi, Dania V.; Burke, Teresa F.; Hensler, Julie G.

    2008-01-01

    The effect of chronic citalopram or escitalopram administration on 5-HT1A receptor function in the dorsal raphe nucleus was determined by measuring [35S]GTPγS binding stimulated by the 5-HT1A receptor agonist (R)-(+)-8-OH-DPAT (1nM-10μM). Although chronic administration of citalopram or escitalopram has been shown to desensitize somatodendritic 5-HT1A autoreceptors, we found that escitalopram treatment decreased the efficacy of 5-HT1A receptors to activate G-proteins, whereas citalopram treat...

  5. Implementation of Nationwide Real-time Whole-genome Sequencing to Enhance Listeriosis Outbreak Detection and Investigation.

    Jackson, Brendan R; Tarr, Cheryl; Strain, Errol; Jackson, Kelly A; Conrad, Amanda; Carleton, Heather; Katz, Lee S; Stroika, Steven; Gould, L Hannah; Mody, Rajal K; Silk, Benjamin J; Beal, Jennifer; Chen, Yi; Timme, Ruth; Doyle, Matthew; Fields, Angela; Wise, Matthew; Tillman, Glenn; Defibaugh-Chavez, Stephanie; Kucerova, Zuzana; Sabol, Ashley; Roache, Katie; Trees, Eija; Simmons, Mustafa; Wasilenko, Jamie; Kubota, Kristy; Pouseele, Hannes; Klimke, William; Besser, John; Brown, Eric; Allard, Marc; Gerner-Smidt, Peter

    2016-08-01

    Listeria monocytogenes (Lm) causes severe foodborne illness (listeriosis). Previous molecular subtyping methods, such as pulsed-field gel electrophoresis (PFGE), were critical in detecting outbreaks that led to food safety improvements and declining incidence, but PFGE provides limited genetic resolution. A multiagency collaboration began performing real-time, whole-genome sequencing (WGS) on all US Lm isolates from patients, food, and the environment in September 2013, posting sequencing data into a public repository. Compared with the year before the project began, WGS, combined with epidemiologic and product trace-back data, detected more listeriosis clusters and solved more outbreaks (2 outbreaks in pre-WGS year, 5 in WGS year 1, and 9 in year 2). Whole-genome multilocus sequence typing and single nucleotide polymorphism analyses provided equivalent phylogenetic relationships relevant to investigations; results were most useful when interpreted in context of epidemiological data. WGS has transformed listeriosis outbreak surveillance and is being implemented for other foodborne pathogens. PMID:27090985

  6. High-throughput sequencing enhanced phage display enables the identification of patient-specific epitope motifs in serum

    Christiansen, Anders; Kringelum, Jens Vindahl; Hansen, Christian Skjødt;

    2015-01-01

    -throughput sequencing. A bioinformatics approach was developed in order to identify peptide motifs of interest based on clustering and contrasting to control samples. Comparison of patient and control samples confirmed a major issue in phage display, namely the selection of unspecific peptides. The potential...... of the bioinformatic approach was demonstrated by identifying epitopes of a prominent peanut allergen, Ara h 1, in sera from patients with severe peanut allergy. The identified epitopes were confirmed by high-density peptide micro-arrays. The present study demonstrates that high-throughput sequencing can empower phage...

  7. Sequences enhancing cassava mosaic disease symptoms occur in the cassava genome and are associated with South African cassava mosaic virus infection.

    Maredza, A T; Allie, F; Plata, G; Rey, M E C

    2016-06-01

    Cassava is an important food security crop in Sub-Saharan Africa. Two episomal begomovirus-associated sequences, named Sequences Enhancing Geminivirus Symptoms (SEGS1 and SEGS2), were identified in field cassava affected by the devastating cassava mosaic disease (CMD). The sequences reportedly exacerbated CMD symptoms in the tolerant cassava landrace TME3, and the model plants Arabidopsis thaliana and Nicotiana benthamiana, when biolistically co-inoculated with African cassava mosaic virus-Cameroon (ACMV-CM) or East African cassava mosaic virus-UG2 (EACMV-UG2). Following the identification of small SEGS fragments in the cassava EST database, the intention of this study was to confirm their presence in the genome, and investigate a possible role for these sequences in CMD. We report that multiple copies of varying lengths of both SEGS1 and SEGS2 are widely distributed in the sequenced cassava genome and are present in several other cassava accessions screened by PCR. The endogenous SEGS1 and SEGS2 are in close proximity or overlapping with cassava genes, suggesting a possible role in regulation of specific biological processes. We confirm the expression of SEGS in planta using EST data and RT-PCR. The sequence features of endogenous SEGS (iSEGS) are unique but resemble non-autonomous transposable elements (TEs) such as MITEs and helitrons. Furthermore, many SEGS-associated genes, some involved in virus-host interactions, are differentially expressed in susceptible (T200) and tolerant TME3) cassava landraces infected by South African cassava mosaic virus (SACMV) of susceptible (T200) and tolerant (TME3) cassava landraces. Abundant SEGS-derived small RNAs were also present in mock-inoculated and SACMV-infected T200 and TME3 leaves. Given the known role of TEs and associated genes in gene regulation and plant immune responses, our observations are consistent with a role of these DNA elements in the host's regulatory response to geminiviruses. PMID:25920485

  8. Effects of source distribution, dose, and linear energy transfer capacity on inactivation and mutation of mycobacteria after 2H, 35S, and 32P incorporation

    Using a selected model, the paper makes a contribution to the question whether the energy dose as a macroscopically-physically defined quantity can be usefully applied in cell ranges with linear dimensions of the order of 1 μm, i.e. whether there is still a correlation between the energy dose and quantitatively measurable biological radiation effects. The problem is investigated with the aid of the intracellular β decay of the 3H, 35S, and 32P nuclei on mycobacteria (BCG) in liquid media. Quantitative findings of radiobiological experiments are linked with model dose calculations to form dose-effect curves. The experimental principle consists in adding radioactively labelled compounds to the nutrient solution of bacteria at normal growth temperatures, thus obtaining an intracellular β source region caused by their uptake. The uptake conditions for the three radionuclides are varied by using different chemical bonds (2H) or carrier concentrations (3H, 35S). As biological reactions, inactivation in the form of growth inhibition and mutagenic induction of resistance to isonicotinic acid hydrazide are recorded. (orig./MG)

  9. Incorporation of 35S-sulfate and 3H-glucosamine into heparan and chondroitin sulfates during the cell cycle of B16-F10 cells

    Changes in glycosaminoglycan composition occurring during the cell cycle were determined in B16-F10 cells sorted flow cytometrically with respect to DNA content. Incorporation of 35S-sulfate into heparan sulfate and chondroitin sulfate of unsorted and G1,S, and G2 +M sorted cells was determined following chondroitinase ABC or nitrous acid treatment; the incorporation into surface material was measured as the difference between the radioactivity of control and trypsin-treated cells. Incorporation of 35S-sulfate and 3H-glucosamine into cetyl pyridinium chloride (CPC)-precipitable material was characterized before and after chondroitinase or nitrous acid treatment by Sephadex G50 chromatography. Long-term (48 h) and short-term (1 h) labeling studies demonstrate that (a) the amount of total cellular chondroitin sulfate is greater than that of heparan sulfate, with larger amounts of unsulfated heparan than chondroitin being present; (b) the rate of turnover of heparan sulfate is greater than that of chondroitin sulfate; (c) greatest short-term incorporation of 3H-glucosamine into CPC-precipitable material occurs during S phase; and (d) the rate of turnover of both heparan sulfate and chondroitin sulfate is decreased in S phase relative to G1 and G2 + M

  10. The specific DNA recognition sequence of the bovine papillomavirus E2 protein is an E2-dependent enhancer.

    Hawley-Nelson, P; Androphy, E J; Lowy, D R; Schiller, J T

    1988-01-01

    The upstream regulatory region (URR) of the bovine papillomavirus (BPV) genome contains an enhancer that is activated by a BPV E2 gene product. We have previously found that a bacterially derived E2 fusion protein specifically interacted with several fragments of URR DNA, suggesting that E2 may activate transcription by directly binding to the enhancer. Each of the bound fragments contains at least one copy of a conserved motif (ACCN6GGT). To determine if this motif is required and sufficient...

  11. Guanine nucleotide-binding proteins that enhance choleragen ADP-ribosyltransferase activity: nucleotide and deduced amino acid sequence of an ADP-ribosylation factor cDNA.

    Price, S R; Nightingale, M.; Tsai, S C; Williamson, K. C.; Adamik, R; H. C. Chen; Moss, J; M. Vaughan

    1988-01-01

    Three (two soluble and one membrane) guanine nucleotide-binding proteins (G proteins) that enhance ADP-ribosylation of the Gs alpha stimulatory subunit of the adenylyl cyclase (EC 4.6.1.1) complex by choleragen have recently been purified from bovine brain. To further define the structure and function of these ADP-ribosylation factors (ARFs), we isolated a cDNA clone (lambda ARF2B) from a bovine retinal library by screening with a mixed heptadecanucleotide probe whose sequence was based on th...

  12. Secure Digital Cashless Transactions with Sequence Diagrams and Spatial Circuits to Enhance the Information Assurance and Security Education

    Dr. Ajantha Herath

    2012-04-01

    Full Text Available Often students have difficulties mastering cryptographic algorithms. For some time we have been developing with methods for introducing important security concepts for both undergraduate and graduate students in Information Systems, Computer Science and Engineering students. To achieve this goal, Sequence diagrams and spatial circuit derivation from equations are introduced to students. Sequence diagrams represent progression of events with time. They learn system security concepts more effectively if they know how to transform equations and high level programming language constructs into spatial circuits or special purpose hardware. This paper describes an active learning module developed to help students understand secure protocols, algorithms and modeling web applications to prevent attacks and both software and hardware implementations related to encryption. These course materials can also be used in computer organization and architecture classes to help students understand and develop special purpose circuitry for cryptographic algorithms.

  13. Enhancing Students Motivation towards School Science with an Inquiry - Based Site Visit Teaching Sequence: A Design - Based Research Approach

    Anni Loukomies

    2013-01-01

    An inquiry-based site visit teaching sequence for school science was designed in co-operation with researchers and science teachers, according to the principles of Design Based Research (DBR). Out-of-school industry site visits were central in the design. Theory-based conjectures arising from the literature on motivation, interest and inquiry-based science teaching (IBST) were embodied in the design solution, and these embodied conjectures were studied in order to uncover the aspects of the d...

  14. Genome Sequence and Transcriptome Analyses of Chrysochromulina tobin: Metabolic Tools for Enhanced Algal Fitness in the Prominent Order Prymnesiales (Haptophyceae).

    Hovde, Blake T; Deodato, Chloe R; Hunsperger, Heather M; Ryken, Scott A; Yost, Will; Jha, Ramesh K; Patterson, Johnathan; Monnat, Raymond J; Barlow, Steven B; Starkenburg, Shawn R; Cattolico, Rose Ann

    2015-01-01

    Haptophytes are recognized as seminal players in aquatic ecosystem function. These algae are important in global carbon sequestration, form destructive harmful blooms, and given their rich fatty acid content, serve as a highly nutritive food source to a broad range of eco-cohorts. Haptophyte dominance in both fresh and marine waters is supported by the mixotrophic nature of many taxa. Despite their importance the nuclear genome sequence of only one haptophyte, Emiliania huxleyi (Isochrysidales), is available. Here we report the draft genome sequence of Chrysochromulina tobin (Prymnesiales), and transcriptome data collected at seven time points over a 24-hour light/dark cycle. The nuclear genome of C. tobin is small (59 Mb), compact (∼ 40% of the genome is protein coding) and encodes approximately 16,777 genes. Genes important to fatty acid synthesis, modification, and catabolism show distinct patterns of expression when monitored over the circadian photoperiod. The C. tobin genome harbors the first hybrid polyketide synthase/non-ribosomal peptide synthase gene complex reported for an algal species, and encodes potential anti-microbial peptides and proteins involved in multidrug and toxic compound extrusion. A new haptophyte xanthorhodopsin was also identified, together with two "red" RuBisCO activases that are shared across many algal lineages. The Chrysochromulina tobin genome sequence provides new information on the evolutionary history, ecology and economic importance of haptophytes. PMID:26397803

  15. Genome Sequence and Transcriptome Analyses of Chrysochromulina tobin: Metabolic Tools for Enhanced Algal Fitness in the Prominent Order Prymnesiales (Haptophyceae.

    Blake T Hovde

    Full Text Available Haptophytes are recognized as seminal players in aquatic ecosystem function. These algae are important in global carbon sequestration, form destructive harmful blooms, and given their rich fatty acid content, serve as a highly nutritive food source to a broad range of eco-cohorts. Haptophyte dominance in both fresh and marine waters is supported by the mixotrophic nature of many taxa. Despite their importance the nuclear genome sequence of only one haptophyte, Emiliania huxleyi (Isochrysidales, is available. Here we report the draft genome sequence of Chrysochromulina tobin (Prymnesiales, and transcriptome data collected at seven time points over a 24-hour light/dark cycle. The nuclear genome of C. tobin is small (59 Mb, compact (∼ 40% of the genome is protein coding and encodes approximately 16,777 genes. Genes important to fatty acid synthesis, modification, and catabolism show distinct patterns of expression when monitored over the circadian photoperiod. The C. tobin genome harbors the first hybrid polyketide synthase/non-ribosomal peptide synthase gene complex reported for an algal species, and encodes potential anti-microbial peptides and proteins involved in multidrug and toxic compound extrusion. A new haptophyte xanthorhodopsin was also identified, together with two "red" RuBisCO activases that are shared across many algal lineages. The Chrysochromulina tobin genome sequence provides new information on the evolutionary history, ecology and economic importance of haptophytes.

  16. 类硅铑离子的3s23p23P1,2-3s3p35S2的跃迁谱线%The Intercombination Lines of 3s23p23P1,2-3s3p35S2 in Si-like Ions, Rhodium

    2002-01-01

    用束箔法研究了类硅铑离子的3s23p23P1,2-3s3p35S2的禁戒跃迁谱线.谱线识别从已知基态精细结构的分裂、基于分支比的强度比、相似的衰减特性、离子束能量下的谱线预期值方面着手.识别后,通过对已知谱线的波长的等电子系列曲线插值或外推来获得用于较刻的谱线的波长,然后较刻出3s23p23P1,2-3s3p35S2这两条谱线.

  17. Application and technical analysis of enhanced T2* star weighted angiography sequence in the detection of hemorrhagic shearing lesions associated with diffuse axonal injury

    Objective: To compare the efficiency of enhanced T2* weighted angiography (ESWAN) sequence with that of a conventional T2* -weighted gradient-recalled-echo (GRE T2* WI) sequence for the detection of hemorrhagic shearing lesions in patients with diffuse axonal injury (DAI). And combined with MRI parameters, to further discuss the principles and virtues of ESWAN sequence. Methods: Seventeen patients with DAI were enrolled in this study. The raw data acquired from ESWAN scan were postprocessed by using the mean square root of multi-echoes. Then, the postprocessed images were compared with the conventional GRE T2* weighted images. The global and regional (superficial, deep and posterior fossa) lesion numbers determined by both sequences were compared by using Wilcoxon signed ranks test (two- tailed). Differences were considered to be significant at P≤0.05. Results: Hemorrhagic lesions were more obvious on ESWAN images than those on conventional GRE T2* weighted images. The median and range value of the detected lesion numbers on ESWAN images were 27 and (1-239) in whole brain, 13 and (1- 89) in cerebral superficial region, 5 and (0-111) in cerebral deep region and 1 and (0-39) in posterior fossa region, respectively; whereas, on GRE T2* weighted images, they were 7 and (1-34) in whole brain, 5 and (1-27) in cerebral superficial region, 2 and (0-25) in cerebral deep region and O and (0-4) in posterior fossa region, respectively. There were significant statistical differences between the two sequences in revealing the lesions in all the four regions (Z=-3.519, -3.182, -3.185, -2.677, P2”* WI and presented more valuable information for the diagnosis of DAI. (authors)

  18. Bioinformatic analysis of neurotropic HIV envelope sequences identifies polymorphisms in the gp120 bridging sheet that increase macrophage-tropism through enhanced interactions with CCR5

    Mefford, Megan E., E-mail: megan_mefford@hms.harvard.edu [Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Boston, MA (United States); Kunstman, Kevin, E-mail: kunstman@northwestern.edu [Northwestern University Medical School, Chicago, IL (United States); Wolinsky, Steven M., E-mail: s-wolinsky@northwestern.edu [Northwestern University Medical School, Chicago, IL (United States); Gabuzda, Dana, E-mail: dana_gabuzda@dfci.harvard.edu [Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Boston, MA (United States); Department of Neurology (Microbiology and Immunobiology), Harvard Medical School, Boston, MA (United States)

    2015-07-15

    Macrophages express low levels of the CD4 receptor compared to T-cells. Macrophage-tropic HIV strains replicating in brain of untreated patients with HIV-associated dementia (HAD) express Envs that are adapted to overcome this restriction through mechanisms that are poorly understood. Here, bioinformatic analysis of env sequence datasets together with functional studies identified polymorphisms in the β3 strand of the HIV gp120 bridging sheet that increase M-tropism. D197, which results in loss of an N-glycan located near the HIV Env trimer apex, was detected in brain in some HAD patients, while position 200 was estimated to be under positive selection. D197 and T/V200 increased fusion and infection of cells expressing low CD4 by enhancing gp120 binding to CCR5. These results identify polymorphisms in the HIV gp120 bridging sheet that overcome the restriction to macrophage infection imposed by low CD4 through enhanced gp120–CCR5 interactions, thereby promoting infection of brain and other macrophage-rich tissues. - Highlights: • We analyze HIV Env sequences and identify amino acids in beta 3 of the gp120 bridging sheet that enhance macrophage tropism. • These amino acids at positions 197 and 200 are present in brain of some patients with HIV-associated dementia. • D197 results in loss of a glycan near the HIV Env trimer apex, which may increase exposure of V3. • These variants may promote infection of macrophages in the brain by enhancing gp120–CCR5 interactions.

  19. Bioinformatic analysis of neurotropic HIV envelope sequences identifies polymorphisms in the gp120 bridging sheet that increase macrophage-tropism through enhanced interactions with CCR5

    Macrophages express low levels of the CD4 receptor compared to T-cells. Macrophage-tropic HIV strains replicating in brain of untreated patients with HIV-associated dementia (HAD) express Envs that are adapted to overcome this restriction through mechanisms that are poorly understood. Here, bioinformatic analysis of env sequence datasets together with functional studies identified polymorphisms in the β3 strand of the HIV gp120 bridging sheet that increase M-tropism. D197, which results in loss of an N-glycan located near the HIV Env trimer apex, was detected in brain in some HAD patients, while position 200 was estimated to be under positive selection. D197 and T/V200 increased fusion and infection of cells expressing low CD4 by enhancing gp120 binding to CCR5. These results identify polymorphisms in the HIV gp120 bridging sheet that overcome the restriction to macrophage infection imposed by low CD4 through enhanced gp120–CCR5 interactions, thereby promoting infection of brain and other macrophage-rich tissues. - Highlights: • We analyze HIV Env sequences and identify amino acids in beta 3 of the gp120 bridging sheet that enhance macrophage tropism. • These amino acids at positions 197 and 200 are present in brain of some patients with HIV-associated dementia. • D197 results in loss of a glycan near the HIV Env trimer apex, which may increase exposure of V3. • These variants may promote infection of macrophages in the brain by enhancing gp120–CCR5 interactions

  20. Disruption of the splicing enhancer sequence within exon 27 of the dystrophin gene by a nonsense mutation induces partial skipping of the exon and is responsible for Becker muscular dystrophy.

    Shiga, N; Takeshima, Y; Sakamoto, H; Inoue, K; Yokota, Y; Yokoyama, M; Matsuo, M

    1997-11-01

    The mechanism of exon skipping induced by nonsense mutations has not been well elucidated. We now report results of in vitro splicing studies which disclosed that a particular example of exon skipping is due to disruption of a splicing enhancer sequence located within the exon. A nonsense mutation (E1211X) due to a G to T transversion at the 28th nucleotide of exon 27 (G3839T) was identified in the dystrophin gene of a Japanese Becker muscular dystrophy case. Partial skipping of the exon resulted in the production of truncated dystrophin mRNA, although the consensus sequences for splicing at both ends of exon 27 were unaltered. To determine how E1211X induced exon 27 skipping, the splicing enhancer activity of purine-rich region within exon 27 was examined in an in vitro splicing system using chimeric doublesex gene pre-mRNA. The mutant sequence containing G3839T abolished splicing enhancer activity of the wild-type purine-rich sequence for the upstream intron in this chimeric pre-mRNA. An artificial polypurine oligonucleotide mimicking the purine-rich sequence of exon 27 also showed enhancer activity that was suppressed by the introduction of a T nucleotide. Furthermore, the splicing enhancer activity was more markedly inhibited when a nonsense codon was created by the inserted T residue. This is the first evidence that partial skipping of an exon harboring a nonsense mutation is due to disruption of a splicing enhancer sequence. PMID:9410897

  1. Radiative-lifetime and Lande-factor measurements of the Se I 4p35s5S2 level using pulsed laser spectroscopy

    First laser spectroscopic investigations on atomic selenium are reported. Natural selenium was thermally dissociated in a quartz resonance cell keeping the background pressure of abundant selenium molecules low by differential heating. The 4p35s5S2 level was excited by frequency-tripled pulsed-dye laser radiation at 207 nm. From time-resolved recordings of the fluorescence decay at 216 nm a natural radiative lifetime of 493(15) ns was determined, while quantum-beat and optical double-resonance measurements in an external magnetic field yielded gJ=2.0004(10) for the Lande factor. The results are compared with previous theoretical and experimental data. (orig.)

  2. Demonstration of the therapeutic effect of /sup 35/S labelled L-cystine in articular and intervertebral cartilage as well as in skeletal musculature

    Schmiegelow, P.; Puschmann, M.; Giese, U.

    1984-01-16

    Clinical experience has obviously shown a positive effect of application of sulfated amino acids on degenerative cartilage diseases. L-Cystin, presumed to be of therapeutic effect, was autoradiographically localized in articular, columnar and intervertebral cartilage as well as in skeletal musculature. In 10 days old NMRI-mice, we had shown a dose-dependent incorporation of the radioactively labelled /sup 35/S-Cystin in hair follicle. These statistically significant differences had been measured by quantitative autoradiographical microscope photometry. The sulfated amino acids are also proven in nail matrix, nail hyponychium as well as in cartilage and skeletal musculature. Besides a localization of radioactively labelled L-Cystin in tissues, presumed as target organs of a therapeutic effect, there is still lacking an experimental proof of efficacy on cell proliferation and functional metabolism e.g. in arthrosis by suitable animal models.

  3. Demonstration of the therapeutic effect of 35S labelled L-cystine in articular and intervertebral cartilage as well as in skeletal musculature

    Clinical experience has obviously shown a positive effect of application of sulfated amino acids on degenerative cartilage diseases. L-Cystin, presumed to be of therapeutic effect, was autoradiographically localized in articular, columnar and intervertebral cartilage as well as in skeletal musculature. In 10 days old NMRI-mice, we had shown a dose-dependent incorporation of the radioactively labelled 35S-Cystin in hair follicle. These statistically significant differences had been measured by quantitative autoradiographical microscope photometry. The sulfated amino acids are also proven in nail matrix, nail hyponychium as well as in cartilage and skeletal musculature. Besides a localization of radioactively labelled L-Cystin in tissues, presumed as target organs of a therapeutic effect, there is still lacking an experimental proof of efficacy on cell proliferation and functional metabolism e.g. in arthrosis by suitable animal models. (orig.)

  4. Combination of low and high resolution sequences in two orientations for dynamic contrast-enhanced MRI of the breast: more than a compromise

    The purpose was to combine T1-weighted 3D gradient echo sequences at low and high spatial resolution (and short and longer acquisition time, respectively) in two orientations without compromising signal/time curve analysis and to evaluate the incremental value of assessing architectural features in high resolution images in dynamic contrast-enhanced MR mammography. T1-weighted 3D-FLASH sequences in a 1.5-T scanner (512 x 256 pixel matrix at high resolution; 256 x 128 pixels at low resolution sequences, 72 slices, 1.7-mm slice thickness, TR 8.8 ms, TE 4.5 ms, flip angle 25 ) were acquired in a special order during a single investigation. Three observers evaluated architectural features of 36 histopathologically proven lesions using high or low resolution images independently. Architectural features of each lesion were assessed by rating on two three-point scales. Kappa statistics verified the decrease of inter-observer variability. All observers improved assessment of architectural features regarding high resolution images in transversal and coronal orientation (observer A: eight positive, three negative corrections; B: 12/5; C: 16/4). Most positive corrections resulted from improved detection of morphologic criteria of malignancy. Mean inter-observer agreement significantly (P<0.05) increased from ''slight'' to ''moderate'' (mean weighted κ increased from 0.185 to 0.422). This protocol at the charge of slightly enlarged time for measurement offers an elegant way to improve analysis of architectural features in MRM. (orig.)

  5. Evaluation of iron colloid-enhanced T2-weighted fast MR imaging of hepatocellular carcinoma. Comparison of SE, TSE and TGSE sequences

    We have applied chondroitin sulfate iron colloid (CSIC) as a contrast agent for MRI in detecting hepatocellular carcinoma (HCC) on conventional spin-echo sequences (SE). In this report, we evaluated CSIC-enhanced T2-weighted fast MR imaging of HCC. MR imaging were performed before and after i.v. administration of CSIC in 15 patients with 46 HCCs. T2-weighted SE (1800/80/2, 210 x 256 matrix), T2-weighted turbo spin-echo (TSE1800) (1800/90/5, echo train length=7, 252 x 256 matrix), TSE (3500/90/5, echo train length=7, 252 x 256 matrix) (TSE7), TSE (3500/99/5, echo train length=11, 242 x 256 matrix) (TSE11) and T2-weighted turbo-gradient spine-echo (TGSE) (4500/108/4, echo train length=33, 252 x 256 matrix) images were compared quantitatively and qualitatively. In all sequences, liver signal-to-noise ratio (SNR) was significantly decreased and lesion-to-liver contrast-to-noise ratio (CNR) was significantly increased after CSIC administration. Although decreased ratio in liver and tumor SNR caused by CSIC was smaller on TSE sequences compared with SE and TGSE, increased ratio in lesion-to-liver CNR was largest on TSE7. Either before or after i.v. administration of CSIC, the number of detectable lesions was largest on TSE7. TSE with used longer TR, TE and decreased echo factor was useful method for CSIC-enhanced abdominal MR imaging. (author)

  6. In trangenic rice, alpha- and beta-tubulin regulatory sequences control GUS amount and distribution through intron mediated enhancement and intron dependent spatial expression.

    Gianì, Silvia; Altana, Andrea; Campanoni, Prisca; Morello, Laura; Breviario, Diego

    2009-04-01

    The genomic upstream sequence of the rice tubulin gene OsTub6 has been cloned, sequenced and characterized. The 5'UTR sequence is interrupted by a 446 bp long leader intron. This feature is shared with two other rice beta-tubulin genes (OsTub4 and OsTub1) that, together with OsTub6, group in the same clade in the evolutionary phylogenetic tree of plant beta-tubulins. Similarly to OsTub4, the leader intron of OsTub6 is capable of sustaining intron mediated enhancement (IME) of gene expression, in transient expression assays. A general picture is drawn for three rice alpha-tubulin and two rice beta-tubulin genes in which the first intron of the coding sequence for the formers and the intron present in the 5'UTR for the latters, are important elements for controlling gene expression. We used OsTua2:GUS, OsTua3:GUS, OsTub4:GUS and OsTub6:GUS chimeric constructs to investigate the in vivo pattern of beta-glucuronidase (GUS) expression in transgenic rice plants. The influence of the regulatory introns on expression patterns was evaluated for two of them, OsTua2 and OsTub4. We have thus characterized distinct patterns of expression attributable to each tubulin isotype and we have shown that the presence of the regulatory intron can greatly influence both the amount and the actual site of expression. We propose the term Intron Dependent Spatial Expression (IDSE) to highlight this latter effect. PMID:18668337

  7. The diagnostic efficacy of quantitative liver MR imaging with diffusion-weighted, SWI, and hepato-specific contrast-enhanced sequences in staging liver fibrosis - a multiparametric approach

    To assess the diagnostic efficacy of multiparametric MRI using quantitative measurements of the apparent diffusion coefficient (ADC) of the liver parenchyma on diffusion-weighted imaging (DWI), signal intensity (SI) on susceptibility-weighted imaging (SWI), and gadoxetic acid-enhanced T1-weighted imaging during the hepatobiliary phase for the staging of liver fibrosis. Seventy-seven patients underwent a 3T MRI examination, including DWI/SWI sequences and gadoxetic acid-enhanced T1-weighted MRI. Liver fibrosis according to liver biopsy was staged using the Metavir fibrosis score: F0 (n = 21, 27.3 %); F1 (n = 7, 9.1 %); F2 (n = 8, 10.4 %); F3 (n = 12, 15.6 %); and F4 (n = 29, 37.7 %). SI of the liver was defined using region-of-interest measurements to calculate the ADC values, the relative enhancement (RE) in the hepatobiliary phase, and the liver-to-muscle ratio (LMR) measurements for SWI. The values of RE, LMR, and ADC measurements were statistically significantly different among the five fibrosis stages (p < 0.004). Combining the three parameters in a multiparametric approach, the AUC for detecting F1 stage or greater (≥ F1) was 94 %, for F2 or greater (≥F2) was 95 %, for F3 or greater (≥F3) was 90 %, and for stage F4 was 93 %. Multiparametric MRI is an efficient non-invasive diagnostic tool for the staging of liver fibrosis. (orig.)

  8. The diagnostic efficacy of quantitative liver MR imaging with diffusion-weighted, SWI, and hepato-specific contrast-enhanced sequences in staging liver fibrosis - a multiparametric approach

    Feier, Diana [Medical University of Vienna, General Hospital of Vienna (AKH), Department of Biomedical Imaging and Image-guided Therapy, Vienna (Austria); Iuliu Hatieganu University of Medicine and Pharmacy, Cluj-Napoca, Emergency County Hospital, Department of Radiology, Cluj-Napoca (Romania); Balassy, Csilla; Bastati, Nina; Fragner, Romana; Ba-Ssalamah, Ahmed [Medical University of Vienna, General Hospital of Vienna (AKH), Department of Biomedical Imaging and Image-guided Therapy, Vienna (Austria); Wrba, Friedrich [Medical University of Vienna, General Hospital of Vienna (AKH), Department of Pathology, Vienna (Austria)

    2016-02-15

    To assess the diagnostic efficacy of multiparametric MRI using quantitative measurements of the apparent diffusion coefficient (ADC) of the liver parenchyma on diffusion-weighted imaging (DWI), signal intensity (SI) on susceptibility-weighted imaging (SWI), and gadoxetic acid-enhanced T1-weighted imaging during the hepatobiliary phase for the staging of liver fibrosis. Seventy-seven patients underwent a 3T MRI examination, including DWI/SWI sequences and gadoxetic acid-enhanced T1-weighted MRI. Liver fibrosis according to liver biopsy was staged using the Metavir fibrosis score: F0 (n = 21, 27.3 %); F1 (n = 7, 9.1 %); F2 (n = 8, 10.4 %); F3 (n = 12, 15.6 %); and F4 (n = 29, 37.7 %). SI of the liver was defined using region-of-interest measurements to calculate the ADC values, the relative enhancement (RE) in the hepatobiliary phase, and the liver-to-muscle ratio (LMR) measurements for SWI. The values of RE, LMR, and ADC measurements were statistically significantly different among the five fibrosis stages (p < 0.004). Combining the three parameters in a multiparametric approach, the AUC for detecting F1 stage or greater (≥ F1) was 94 %, for F2 or greater (≥F2) was 95 %, for F3 or greater (≥F3) was 90 %, and for stage F4 was 93 %. Multiparametric MRI is an efficient non-invasive diagnostic tool for the staging of liver fibrosis. (orig.)

  9. Studies of artifacts with TFE factor increase in heart delayed enhancement MRI sequence IR-T1TFE

    The characteristic of delayed enhancement MRI is high spatial resolution, which makes it possible to evaluate the degree of damage to the myocardium from the inner to the outer membrane of patients with ischemic heart disease. Therefore, this MRI technique is unique in its ability to detect myocardial condition and is necessary to obtain high-quality images. We experienced artifacts induced by turbo field echo (TFE) factor increase with delayed enhancement of inversion recovery-T1 TFE (IR-T1TFE). The purpose of this study was to determine the cause of such artifacts. IR-T1TFE changed signal intensity with phase direction as the TFE factor increased. Streak artifacts occurred because signal intensity caused changes with phase direction. Increases in TFE factor prolonged data collection time, such that marked artifacts were created because of changes in signal intensity. Ghost artifacts occurred because signal intensity changed between shots. When the TFE factor was increased, the difference in signal intensity was diminished between shots. The interval of acquired noise decreased in the raw data. Therefore, the interval of ghost artifacts became wider on images. (author)

  10. Enhanced Biological Phosphorus Removal in Anaerobic/Aerobic Sequencing Batch Reactor Supplied with Glucose as Carbon Source

    LIU Yanan; YU Shui-li; JING Guo-lin; ZHAO Bing-jie; GUO Si-yuan

    2005-01-01

    Phosphorus removal performance in an aerobic/aerobic sequencing batch reactor (SBR) supplied with glucose as carbon source was investigated. It was found that there was no phosphate release concomitant with the storing of poly-β-hydroxyalkanoate (PHA) during the anaerobic phase. Whereas, glycogen was soon built up followed by rapid consumption, at the same time, glucose was depleted rapidly. Based on the analysis of different fractions of phosphorus in activated sludge, the relative ratio of organically bound phosphorus in sludge changed at the end of anaerobic and aerobic phases. The ratios were 45.3% and51.8% respectively. This showed that the polyphosphate broke down during the anaerobic phase to supply part of energy for PHA synthesis. The reason why there was no phosphate release might be the biosorption effect of extracellular exopolymers (EPS). It was also proved by the analysis of EPS with scanning electron microscopy (SEM)combined with energy dispersive spectrometry (EDS). The phosphorus weight percentage of EPS at the end of anaerobic phase was 9.22%.

  11. Combination of low and high resolution sequences in two orientations for dynamic contrast-enhanced MRI of the breast: more than a compromise

    Vomweg, Toni W.; Teifke, Andrea; Kunz, R. Peter; Hintze, Christian; Hlawatsch, Alexander; Kern, Annett; Kreitner, Karl F.; Thelen, Manfred [Johannes Gutenberg University, Department of Radiology, University Medical Centre, Mainz (Germany)

    2004-10-01

    The purpose was to combine T1-weighted 3D gradient echo sequences at low and high spatial resolution (and short and longer acquisition time, respectively) in two orientations without compromising signal/time curve analysis and to evaluate the incremental value of assessing architectural features in high resolution images in dynamic contrast-enhanced MR mammography. T1-weighted 3D-FLASH sequences in a 1.5-T scanner (512 x 256 pixel matrix at high resolution; 256 x 128 pixels at low resolution sequences, 72 slices, 1.7-mm slice thickness, TR 8.8 ms, TE 4.5 ms, flip angle 25 ) were acquired in a special order during a single investigation. Three observers evaluated architectural features of 36 histopathologically proven lesions using high or low resolution images independently. Architectural features of each lesion were assessed by rating on two three-point scales. Kappa statistics verified the decrease of inter-observer variability. All observers improved assessment of architectural features regarding high resolution images in transversal and coronal orientation (observer A: eight positive, three negative corrections; B: 12/5; C: 16/4). Most positive corrections resulted from improved detection of morphologic criteria of malignancy. Mean inter-observer agreement significantly (P<0.05) increased from ''slight'' to ''moderate'' (mean weighted {kappa} increased from 0.185 to 0.422). This protocol at the charge of slightly enlarged time for measurement offers an elegant way to improve analysis of architectural features in MRM. (orig.)

  12. Morpholino spin-labeling for base-pair sequencing of a 3'-terminal RNA stem by proton homonuclear Overhauser enhancements: yeast ribosomal 5S RNA

    Base-pair sequences for 5S and 5.8S RNAs are not readily extracted from proton homonuclear nuclear Overhauser enhancement (NOE) connectivity experiments alone, due to extensive peak overlap in the downfield (11-15 ppm) proton NMR spectrum. In this paper, we introduce a new method for base-pair proton peak assignment for ribosomal RNAs, based upon the distance-dependent broadening of the resonances of base-pair protons spatially proximal to a paramagnetic group. Introduction of a nitroxide spin-label covalently attached to the 3'-terminal ribose provides an unequivocal starting point for base-pair hydrogen-bond proton NMR assignment. Subsequent NOE connectivities then establish the base-pair sequence for the terminal stem of a 5S RNA. Periodate oxidation of yeast 5S RNA, followed by reaction with 4-amino-2,2,6,6-tetramethylpiperidinyl-1-oxy (TEMPO-NH2) and sodium borohydride reduction, produces yeast 5S RNA specifically labeled with a paramagnetic nitroxide group at the 3'-terminal ribose. Comparison of the 500-MHz 1H NMR spectra of native and 3'-terminal spin-labeled yeast 5S RNA serves to identify the terminal base pair (G1 . C120) and its adjacent base pair (G2 . U119) on the basis of their proximity to the 3'-terminal spin-label. From that starting point, we have then identified (G . C, A . U, or G . U) and sequenced eight of the nine base pairs in the terminal helix via primary and secondary NOE's

  13. Assessment of myocardial infarction in mice by Late Gadolinium Enhancement MR imaging using an inversion recovery pulse sequence at 9.4T

    Herlihy Amy H

    2008-01-01

    Full Text Available Abstract Purpose To demonstrate the feasibility of using an inversion recovery pulse sequence and to define the optimal inversion time (TI to assess myocardial infarction in mice by late gadolinium enhancement (LGE MRI at 9.4T, and to obtain the maximal contrast between the infarcted and the viable myocardium. Methods MRI was performed at 9.4T in mice, two days after induction of myocardial infarction (n = 4. For cardiovascular MR imaging, a segmented magnetization-prepared fast low angle shot (MP-FLASH sequence was used with varied TIs ranging from 40 to 420 ms following administration of gadolinium-DTPA at 0.6 mmol/kg. Contrast-to-noise (CNR and signal-to-noise ratio (SNR were measured and compared for each myocardial region of interest (ROI. Results The optimal TI, which corresponded to a minimum SNR in the normal myocardium, was 268 ms ± 27.3. The SNR in the viable myocardium was significantly different from that found in the infarcted myocardium (17.2 ± 2.4 vs 82.1 ± 10.8; p = 0.006 leading to a maximal relative SI (Signal Intensity between those two areas (344.9 ± 60.4. Conclusion Despite the rapid heart rate in mice, our study demonstrates that LGE MRI can be performed at 9.4T using a protocol similar to the one used for clinical MR diagnosis of myocardial infarction.

  14. Fusion of a Short HA2-Derived Peptide Sequence to Cell-Penetrating Peptides Improves Cytosolic Uptake, but Enhances Cytotoxic Activity

    Igor Kitanovic

    2009-09-01

    Full Text Available Cell-penetrating peptides (CPP have become a widely used tool for efficient cargo delivery into cells. However, one limiting fact is their uptake by endocytosis causing the enclosure of the CPP-cargo construct within endosomes. One often used method to enhance the outflow into the cytosol is the fusion of endosome-disruptive peptide or protein sequences to CPP. But, until now, no studies exist investigating the effects of the fusion peptide to the cellular distribution, structural arrangements and cytotoxic behaviour of the CPP. In this study, we attached a short modified sequence of hemagglutinin subunit HA2 to different CPP and analysed the biologic activity of the new designed peptides. Interestingly, we observed an increased cytosolic distribution but also highly toxic activities in the micromolar range against several cell lines. Structural analysis revealed that attachment of the fusion peptide had profound implications on the whole conformation of the peptide, which might be responsible for membrane interaction and endosome disruption.

  15. Effect of colchicine and vinblastine over the odontoblasts secretion from mice lower incisor as revealed by autoradiography after injection of 35S-sodium sulphate

    For the biosynthesis study of sulphated compounds in the odontoblasts under the effect of colchicine and vinblastine, male mice, (M. musculus, albinus) weighing 22 to 25 g. were used. They were divided in 3 groups of 3 animals each. One group was used as control, the other received 0.1 mg of colchicine and the last one received 0.2 mg of vinblastine. Two hours after the administration of colchicine and 3 hours after the vinblastine, all the treated animals and the control group received one single dose of 10μCi/g of 35S-sodium sulphate and were sacrificed after 10, 30 and 120 minutes. The lower incisors were prepared for radioautography. This study showed that the secretory odontoblasts in the presence of colchicine and vinblastine, incorporated the radioactive material, but there was a delay in the biosynthesis process of sulphated compounds accumulating the secretory material, which could be due to: 1. the colchicine in the odontoblasts would act in the final fase of the sulphatation process; 2. the vinblastine would act primarily in the local were the proteic substract of the sulphated compounds (glycosaminoglicans) is synthesized, resulting in a lack of material to bind the radiactive sulphur. (Author)

  16. Effect of colchicine and vinblastine over the odontoblasts secretion from mice lower incisor as revealed by autoradiography after injection of /sup 35/S-sodium sulphate

    Azevedo, L.F.; Blumen, G. (Universidade Estadual de Campinas, Piracicaba (Brazil). Faculdade de Odontologia)

    1983-07-01

    For the biosynthesis study of sulphated compounds in the odontoblasts under the effect of colchicine and vinblastine, male mice, (M. musculus, albinus) weighing 22 to 25 g. were used. They were divided in 3 groups of 3 animals each. One group was used as control, the other received 0.1 mg of colchicine and the last one received 0.2 mg of vinblastine. Two hours after the administration of colchicine and 3 hours after the vinblastine, all the treated animals and the control group received one single dose of 10..mu..Ci/g of /sup 35/S-sodium sulphate and were sacrificed after 10, 30 and 120 minutes. The lower incisors were prepared for radioautography. This study showed that the secretory odontoblasts in the presence of colchicine and vinblastine, incorporated the radioactive material, but there was a delay in the biosynthesis process of sulphated compounds accumulating the secretory material, which could be due to: 1. the colchicine in the odontoblasts would act in the final phase of the sulphatation process; 2. the vinblastine would act primarily in the local were the proteic substract of the sulphated compounds (glycosaminoglicans) is synthesized, resulting in a lack of material to bind the radiactive sulphur.

  17. Synthesis of proteins from [35S]methionine by guinea pig megakaryocytes in vivo and time course of appearance of newly synthesized proteins in platelets

    The relationship of protein synthesis to megakaryocyte maturation has been studied in guinea pigs in vivo. Guinea pigs were injected with a single dose of [35S]methionine. Megakaryocytes and platelets were isolated daily for 4 days, and proteins from both cells were isolated by DEAE-Sephacel chromatography and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and fluorography. All proteins in megakaryocytes corresponding to stained bands on the SDS-PAGE gels were radiolabeled at 3 hours after injection. The greatest loss of radioactivity from the megakaryocytes occurred between 1 and 3 days after injection. Only trace labeling of platelet proteins was seen at 3 hours, representing almost entirely three bands at molecular weights 47,000, 52,000, and 66,000. At 24 hours only about 13% of the maximal labeling was present, but not all proteins were labeled. The maximal labeling was at 3 days. The pattern of labeling of platelets at 3 days was identical to that of megakaryocytes at 3 hours. The protein pattern of nonmegakaryocytic marrow cells was different from that of the platelets and megakaryocytes. Data presented here suggest that most protein synthesis in megakaryocytes is completed at least 24 hours before release of the platelets to the circulation, and suggest some specificity in the proteins that are synthesized at the terminal stages of maturation

  18. Distribution of radioactively labelled myobacterium tuberculosis (65Zn, 35S-BCG) after injection into the anterior chamber. Studies in rabbits with and without anterior chamber sensitisation

    The rabbit model was used to study the behaviour of antigens in the eye as well as their spread to extraocular regions. Prior to the investigations, a test dose of 35S-labelled and 65Zn-labelled BCG (Bacille Calmette Guerin) had been injected into the anterior chamber of the eyes of sensitised and non-sensitised rabbits. The percentage of BCG escaping from the aqueous humor and iris was mostly not dependent of the test dose given, nor did the rate of its spread show any relation to the occurrence of an antigen-antibody reaction. On the 32nd day p.i., there were still acid-resisting rods in the uvea, which could be proven by histological examination. As shown by the levels of activity in the iris and ciliary body, the bacteria appear to migrate mostly to these organs and, in the second place, to the choroid membrane. The spread of bacteria in previously sensitised rabbits was not different from the observed in non-sensitised animals. Theories claiming that more antigens are formed in sensitised tissue as compared to non-sensitised tissue cannot be confirmed by the findings revealed here. (TRV)

  19. Concentration dependence of rapid axonal transport: a study of the transport kinetics of [35S]methionine-labeled protein in postganglionic sympathetic fibers of the bullfrog

    The kinetics of transport of radiolabeled proteins in sympathetic axons of the bullfrog sciatic nerve were examined after injection of [35S]methionine into the S9 sympathetic ganglion. Under resting conditions at 20 degrees C, the fastest moving material was carried distally at 5.7 +/- 0.3 mm/hr. Various manipulations of temperature in the proximal part of the nerve were used to alter the amount of protein transported into the distal region, which was always kept at 20 degrees C. The velocity in this test region was found to increase to over 9 mm/hr when material that had accumulated at a cold block for 4 hr was released by rewarming. This acceleration was transient, and base line velocity was regained after 2 hr. In order to increase the local concentration of transported protein by a second method, the proximal part of several nerves was warmed to 28 degrees C. Maximal transport velocity in the 20 degrees C test region rose to 6.2 +/- 0.12 mm/hr. To decrease the local concentration of transported protein, the proximal part of other nerves was cooled to 15 degrees C. Maximal transport velocity in the 20 degrees C test region fell to 4.7 +/- 0.7 mm/hr. We conclude that there is a small but real tendency for the velocity of rapid axonal transport in this neural system to be positively related to the availability of material suitable for transport

  20. Identification of adenovirus type 12 candidate transformation proteins by radioimmunoprecipitation with antisera to EcoRI-C-fragment. [/sup 35/S tracer technique, rats

    Wold, W.S.M. (St. Louis Univ. School of Medicine, MO); Chinnadurai, G.; Green, M.; Mak, S.

    1979-04-15

    Experiments were performed to identify polypeptides coded by early gene block 1 (which includes the transforming region) of human adenoviruses in group A (Ad12, 18, 31). Two lines, C-1 and C-2, of rat cells transformed by transfection with Ad12 EcoRI-C fragment (left 16% of genome) were inoculated into syngeneic rats to produce tumors (F. Graham and S. Mak, unpublished data). The tumor sera were used to immunoprecipitate (/sup 35/S)methionine-labeled polypeptides from Ad12-early-infected human cells. The polypeptides were resolved by electrophoresis in sodium dodecyl sulfate-polyacrylamide gels, and visualized by fluorography. Two additional sera were also used, from hamsters bearing tumors induced by inoculation with Ad12 or Ad18 virions. The immunoprecipitation results suggest (but do not prove) that early gene block 1 of group A Ads may code a family of related polypeptides with apparent molecular weights ranging from 4OK to 46K or 4OK to 65K, as well as polypeptides of 16.5 K and 10.5K. One or more of these polypeptides may play a role in the initiation and/or maintenance of cell transformation.

  1. Efficiency of Non-Contrast-Enhanced Liver Imaging Sequences Added to Initial Rectal MRI in Rectal Cancer Patients.

    Gene-hyuk Kwon

    Full Text Available The purpose of this study was to estimate the value of addition of liver imaging to initial rectal magnetic resonance imaging (MRI for detection of liver metastasis and evaluate imaging predictors of a high risk of liver metastasis on rectal MRI.We enrolled 144 patients who from October 2010 to May 2013 underwent rectal MRI with T2-weighted imaging (T2WI and diffusion-weighted imaging (DWI (b values = 50, 500, and 900 s/mm2 of the liver and abdominopelvic computed tomography (APCT for the initial staging of rectal cancer. Two reviewers scored the possibility of liver metastasis on different sets of liver images (T2WI, DWI, and combined T2WI and DWI and APCT and reached a conclusion by consensus for different analytic results. Imaging features from rectal MRI were also analyzed. The diagnostic performances of CT and an additional liver scan to detect liver metastasis were compared. Multivariate logistic regression to determine independent predictors of liver metastasis among rectal MRI features and tumor markers was performed. This retrospective study was approved by the Institutional Review Board, and the requirement for informed consent was waived.All sets of liver images were more effective than APCT for detecting liver metastasis, and DWI was the most effective. Perivascular stranding and anal sphincter invasion were statistically significant for liver metastasis (p = 0.0077 and p = 0.0471, while extramural vascular invasion based on MRI (mrEMVI was marginally significant (p = 0.0534.The addition of non-contrast-enhanced liver imaging, particularly DWI, to initial rectal MRI in rectal cancer patients could facilitate detection of liver metastasis without APCT. Perivascular stranding, anal sphincter invasion, and mrEMVI detected on rectal MRI were important imaging predictors of liver metastasis.

  2. Enhancement of the performance of an anaerobic sequencing batch reactor treating low-strength wastewater through implementation of a variable stirring rate program

    Rodrigues J. A. D.

    2004-01-01

    Full Text Available This work focuses on enhancement of the performance of an anaerobic sequencing batch reactor with a six-vertical-blade-disk-turbine impeller, containing granulated biomass treating low-strength synthetic wastewater, through a study of the feasibility of implementing a variable stirring rate program. The reactor was operated at 30ºC and a six-hour cycle was used to treat approximately 2.0 L of the synthetic substrate with a chemical oxygen demand (COD of nearly 500 mg/L. Two different stirring rate program were implemented: a constant rate of 50 rpm and a variable rate consisting of 75 rpm for one hour, 50 rpm for four hours and 25 rpm for 0.5 hour. The last 0.5 hour of the cycle was used for the settling step. In both cases, a very short start-up period and unfiltered and filtered substrate removal efficiencies of 81% and 88%, respectively, were attained. However, use of the variable stirring rate enhanced efficiency of the reactor dynamics without impairing biomass morphology, thus resulting in a reduction in the total cycle time and a possible decrease in energy consumption. Additionally, a simplified model of the anaerobic metabolic activity, using apparent kinetic parameters, was proposed as a consecutive first-order kinetic model with substrate and total volatile acid residual concentrations in order to analyze how the variable stirring rate affects reactor performance.

  3. New insights on the transcriptional regulation of CD69 gene through a potent enhancer located in the conserved non-coding sequence 2.

    Laguna, Teresa; Notario, Laura; Pippa, Raffaella; Fontela, Miguel G; Vázquez, Berta N; Maicas, Miren; Aguilera-Montilla, Noemí; Corbí, Ángel L; Odero, María D; Lauzurica, Pilar

    2015-08-01

    The CD69 type II C-type lectin is one of the earliest indicators of leukocyte activation acting in lymphocyte migration and cytokine secretion. CD69 expression in hematopoietic lineage undergoes rapid changes depending on the cell-lineage, the activation state or the localization of the cell where it is expressed, suggesting a complex and tightly controlled regulation. Here we provide new insights on the transcriptional regulation of CD69 gene in mammal species. Through in silico studies, we analyzed several regulatory features of the 4 upstream conserved non-coding sequences (CNS 1-4) previously described, confirming a major function of CNS2 in the transcriptional regulation of CD69. In addition, multiple transcription binding sites are identified in the CNS2 region by DNA cross-species conservation analysis. By functional approaches we defined a core region of 226bp located within CNS2 as the main enhancer element of CD69 transcription in the hematopoietic cells analyzed. By chromatin immunoprecipitation, binding of RUNX1 to the core-CNS2 was shown in a T cell line. In addition, we found an activating but not essential role of RUNX1 in CD69 gene transcription by site-directed mutagenesis and RNA silencing, probably through the interaction with this potent enhancer specifically in the hematopoietic lineage. In summary, in this study we contribute with new evidences to the landscape of the transcriptional regulation of the CD69 gene. PMID:25801305

  4. The HB22.7 Anti-CD22 monoclonal antibody enhances bortezomib-mediated lymphomacidal activity in a sequence dependent manner

    Martin Shiloh M

    2011-12-01

    Full Text Available Abstract Most non-Hodgkin's lymphomas (NHL initially respond to chemotherapy, but relapse is common and treatment is often limited by chemotherapy-related toxicity. Bortezomib, is a highly selective proteasome inhibitor with anti-NHL activity; it is currently FDA approved for second-line treatment of mantle cell lymphoma (MCL. Bortezomib exerts its activity in part through the generation of reactive oxygen species (ROS and also by the induction of apoptosis. We previously validated CD22 as a potential target in treating NHL and have shown that the anti-CD22 ligand blocking antibody, HB22.7, has significant independent lymphomacidal properties in NHL xenograft models. We sought to determine whether or not these agents would work synergistically to enhance cytotoxicity. Our results indicate that treatment of NHL cell lines with HB22.7 six hours prior to bortezomib significantly diminished cell viability. These effects were not seen when the agents were administered alone or when bortezomib was administered prior to HB22.7. Additionally, HB22.7 treatment prior to bortezomib increased apoptosis in part through enhanced ROS generation. Finally, in a mouse xenograft model, administration of HB22.7 followed 24 hours later by bortezomib resulted in 23% smaller tumor volumes and 20% enhanced survival compared to treatment with the reverse sequence. Despite the increased efficacy of HB22.7 treatment followed by bortezomib, there was no corresponding decrease in peripheral blood cell counts, indicating no increase in toxicity. Our results suggest that pre-treatment with HB22.7 increases bortezomib cytotoxicity, in part through increased reactive oxygen species and apoptosis, and that this sequential treatment combination has robust efficacy in vivo.

  5. Tissue distribution of 35S-labelled perfluorooctane sulfonate in adult mice after oral exposure to a low environmentally relevant dose or a high experimental dose

    The widespread environmental pollutant perfluorooctane sulfonate (PFOS), detected in most animal species including the general human population, exerts several effects on experimental animals, e.g., hepatotoxicity, immunotoxicity and developmental toxicity. However, detailed information on the tissue distribution of PFOS in mammals is scarce and, in particular, the lack of available information regarding environmentally relevant exposure levels limits our understanding of how mammals (including humans) may be affected. Accordingly, we characterized the tissue distribution of this compound in mice, an important experimental animal for studying PFOS toxicity. Following dietary exposure of adult male C57/BL6 mice for 1-5 days to an environmentally relevant (0.031 mg/kg/day) or a 750-fold higher experimentally relevant dose (23 mg/kg/day) of 35S-PFOS, most of the radioactivity administered was recovered in liver, bone (bone marrow), blood, skin and muscle, with the highest levels detected in liver, lung, blood, kidney and bone (bone marrow). Following high daily dose exposure, PFOS exhibited a different distribution profile than with low daily dose exposure, which indicated a shift in distribution from the blood to the tissues with increasing dose. Both scintillation counting (with correction for the blood present in the tissues) and whole-body autoradiography revealed the presence of PFOS in all 19 tissues examined, with identification of thymus as a novel site for localization for PFOS and bone (bone marrow), skin and muscle as significant body compartments for PFOS. These findings demonstrate that PFOS leaves the bloodstream and enters most tissues in a dose-dependent manner.

  6. Label-Free Detection of Sequence-Specific DNA Based on Fluorescent Silver Nanoclusters-Assisted Surface Plasmon-Enhanced Energy Transfer.

    Ma, Jin-Liang; Yin, Bin-Cheng; Le, Huynh-Nhu; Ye, Bang-Ce

    2015-06-17

    We have developed a label-free method for sequence-specific DNA detection based on surface plasmon enhanced energy transfer (SPEET) process between fluorescent DNA/AgNC string and gold nanoparticles (AuNPs). DNA/AgNC string, prepared by a single-stranded DNA template encoded two emitter-nucleation sequences at its termini and an oligo spacer in the middle, was rationally designed to produce bright fluorescence emission. The proposed method takes advantage of two strategies. The first one is the difference in binding properties of single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) toward AuNPs. The second one is SPEET process between fluorescent DNA/AgNC string and AuNPs, in which fluorescent DNA/AgNC string can be spontaneously adsorbed onto the surface of AuNPs and correspondingly AuNPs serve as "nanoquencher" to quench the fluorescence of DNA/AgNC string. In the presence of target DNA, the sensing probe hybridized with target DNA to form duplex DNA, leading to a salt-induced AuNP aggregation and subsequently weakened SPEET process between fluorescent DNA/AgNC string and AuNPs. A red-to-blue color change of AuNPs and a concomitant fluorescence increase were clearly observed in the sensing system, which had a concentration dependent manner with specific DNA. The proposed method achieved a detection limit of ∼2.5 nM, offering the following merits of simple design, convenient operation, and low experimental cost because of no chemical modification, organic dye, enzymatic reaction, or separation procedure involved. PMID:26024337

  7. Enhanced Gene Expression Rather than Natural Polymorphism in Coding Sequence of the OsbZIP23 Determines Drought Tolerance and Yield Improvement in Rice Genotypes.

    Avishek Dey

    Full Text Available Drought is one of the major limiting factors for productivity of crops including rice (Oryza sativa L.. Understanding the role of allelic variations of key regulatory genes involved in stress-tolerance is essential for developing an effective strategy to combat drought. The bZIP transcription factors play a crucial role in abiotic-stress adaptation in plants via abscisic acid (ABA signaling pathway. The present study aimed to search for allelic polymorphism in the OsbZIP23 gene across selected drought-tolerant and drought-sensitive rice genotypes, and to characterize the new allele through overexpression (OE and gene-silencing (RNAi. Analyses of the coding DNA sequence (CDS of the cloned OsbZIP23 gene revealed single nucleotide polymorphism at four places and a 15-nucleotide deletion at one place. The single-copy OsbZIP23 gene is expressed at relatively higher level in leaf tissues of drought-tolerant genotypes, and its abundance is more in reproductive stage. Cloning and sequence analyses of the OsbZIP23-promoter from drought-tolerant O. rufipogon and drought-sensitive IR20 cultivar showed variation in the number of stress-responsive cis-elements and a 35-nucleotide deletion at 5'-UTR in IR20. Analysis of the GFP reporter gene function revealed that the promoter activity of O. rufipogon is comparatively higher than that of IR20. The overexpression of any of the two polymorphic forms (1083 bp and 1068 bp CDS of OsbZIP23 improved drought tolerance and yield-related traits significantly by retaining higher content of cellular water, soluble sugar and proline; and exhibited decrease in membrane lipid peroxidation in comparison to RNAi lines and non-transgenic plants. The OE lines showed higher expression of target genes-OsRab16B, OsRab21 and OsLEA3-1 and increased ABA sensitivity; indicating that OsbZIP23 is a positive transcriptional-regulator of the ABA-signaling pathway. Taken together, the present study concludes that the enhanced gene

  8. Enhanced Gene Expression Rather than Natural Polymorphism in Coding Sequence of the OsbZIP23 Determines Drought Tolerance and Yield Improvement in Rice Genotypes.

    Dey, Avishek; Samanta, Milan Kumar; Gayen, Srimonta; Sen, Soumitra K; Maiti, Mrinal K

    2016-01-01

    Drought is one of the major limiting factors for productivity of crops including rice (Oryza sativa L.). Understanding the role of allelic variations of key regulatory genes involved in stress-tolerance is essential for developing an effective strategy to combat drought. The bZIP transcription factors play a crucial role in abiotic-stress adaptation in plants via abscisic acid (ABA) signaling pathway. The present study aimed to search for allelic polymorphism in the OsbZIP23 gene across selected drought-tolerant and drought-sensitive rice genotypes, and to characterize the new allele through overexpression (OE) and gene-silencing (RNAi). Analyses of the coding DNA sequence (CDS) of the cloned OsbZIP23 gene revealed single nucleotide polymorphism at four places and a 15-nucleotide deletion at one place. The single-copy OsbZIP23 gene is expressed at relatively higher level in leaf tissues of drought-tolerant genotypes, and its abundance is more in reproductive stage. Cloning and sequence analyses of the OsbZIP23-promoter from drought-tolerant O. rufipogon and drought-sensitive IR20 cultivar showed variation in the number of stress-responsive cis-elements and a 35-nucleotide deletion at 5'-UTR in IR20. Analysis of the GFP reporter gene function revealed that the promoter activity of O. rufipogon is comparatively higher than that of IR20. The overexpression of any of the two polymorphic forms (1083 bp and 1068 bp CDS) of OsbZIP23 improved drought tolerance and yield-related traits significantly by retaining higher content of cellular water, soluble sugar and proline; and exhibited decrease in membrane lipid peroxidation in comparison to RNAi lines and non-transgenic plants. The OE lines showed higher expression of target genes-OsRab16B, OsRab21 and OsLEA3-1 and increased ABA sensitivity; indicating that OsbZIP23 is a positive transcriptional-regulator of the ABA-signaling pathway. Taken together, the present study concludes that the enhanced gene expression rather than

  9. Using invasion process theory to enhance the understanding and management of introduced species: a case study reconstructing the invasion sequence of the common myna (Acridotheres tristis).

    Grarock, Kate; Lindenmayer, David B; Wood, Jeff T; Tidemann, Chris R

    2013-11-15

    Research indicates that invasion is a multi-step process, where each stage is contingent on the stage that precedes it. Numerous hypotheses addressing the factors that influence each stage of the invasion process have been formulated, but how well does this theory match what occurs in the natural world? We created a general conceptual model for the invasion process based on invasion theory. Using a composite 41-year data set, we then reconstructed the invasion sequence of the common myna (Acridotheres tristis) to investigate the similarities between invasion theory and this observed invasion. We observed a lag period before population growth of 2.7 (±0.3) years, a maximum rate of population growth of 24.1 (±6.4) birds per km(2) per year, a lag period before spreading of six years and an average spreading rate of 0.4 km per year. The length and duration of these stages correspond closely with what invasion process theory would anticipate. We suggest that a conceptual model, coupled with basic species, environment and event information, could be a useful tool to enhance the understanding and management of invasions. PMID:23995141

  10. Expression of 5 S rRNA genes linked to 35 S rDNA in plants, their epigenetic modification and regulatory element divergence

    Garcia, S.; Crhák Khaitová, Lucie; Kovařík, Aleš

    2012-01-01

    Roč. 12, JUN 20 (2012), ID 95. ISSN 1471-2229 R&D Projects: GA ČR GAP501/10/0208; GA ČR GBP501/12/G090 Institutional support: RVO:68081707 Keywords : ARABIDOPSIS-THALIANA * NUCLEOTIDE-SEQUENCES * DNA METHYLATION * GENUS ARTEMISIA Subject RIV: BO - Biophysics Impact factor: 4.354, year: 2012