bluuebunny/crossref_metadata_2025 huggingface dataset:
Lexical Priming and Sound-to-Spelling Contingency Effects in Nonword Spelling
Authors: Barry Christopher , Seymour Philip H. K.
Published: September 2011

Campbell (1983) demonstrated that nonword spelling may be influenced by the spelling patterns of previously heard, rhyming words (“lexical priming”). We report an experiment that compares two nonword spelling tasks: an experimental (“priming”) task, in which nonwords were preceded by rhyming words of different spellings (as in Campbell's task), and a free-spelling task in which only nonwords are presented. The frequency of production of critical spelling patterns was significantly greater in the experimental task than in the free-spelling task (a lexical priming effect). However, there were, and equally for both tasks, significant and substantial effects of sound-to-spelling contingency (i.e. the frequency with which spelling patterns represent vowel phonemes in words): subjects produced more high-contingency (i.e. common) spelling patterns of vowels than low-contingency (rare) spellings. Further, within high-contingency spelling patterns, subjects more frequently produced the most common spelling correspondence of vowels than the second most common spelling. The results are interpreted within a proposed model of assembled spelling, in which it is suggested that there exist a set of probabilistic sound-to-spelling mappings that relate vowel phonemes to weighted lists of alternative spelling patterns ordered by sound-to-spelling contingency, but that the selection of a spelling pattern from such lists is open to lexical influence.
Deafness, Spelling and Rhyme: How Spelling Supports Written Word and Picture Rhyming Skills in Deaf Subjects
Authors: Campbell Ruth , Wright Helen
Published: September 2011

Orally trained, congenitally deaf adolescents and hearing, reading-age-matched control subjects made rhyme judgements for pictures and for written words. Hearing children performed the task accurately. By contrast, the deaf group were very poor at rhyme judgement for words and for pictures. For hearing children, word rhyme judgement was more accurate when the words were congruent in their spelling pattern (e.g. bat/hat), less accurate when the spelling pattern of the rhyming words was incongruent ( hair/bear). Deaf subjects showed an even more pronounced effect of spelling congruence; their ability to match for rhyme when written words did not share the same spelling pattern was extremely poor. Moreover, spelling congruence predicted deaf subjects’ picture rhyming skills.

We conclude that oral training for deaf people does not always permit them to achieve a reliable phonological representation of speech from lip-reading and residual hearing alone. Instead they use the written spelling of the word. This result is not predicted from some previous results that suggest that orally trained deaf people can make direct, spontaneous use of rhyme in the processing of visually presented material.
Unimpaired Short-term Memory and Rhyme Judgement in Congenitally Speechless Individuals: Implications for the Notion of “Articulatory Coding”
Authors: Bishop D. V. M. , Robson J.
Published: September 2011

In normal adults, concurrent articulation impairs short-term memory, abolishing both the phonological similarity effect and the word length effect when visual presentation is used. It also interferes with ability to judge whether visually presented words rhyme. It is generally assumed that concurrent articulation impairs performance because it prevents people from recoding material into an articulatory form. If this is the explanation, then individuals who are congenitally speechless (anarthric) or speech-impaired (dysarthric) should show the same impairments as normal individuals who are concurrently articulating—i.e. they should have reduced memory spans, fail to show word length and phonological similarity effects in short-term memory, and find rhyme judgement difficult. These predictions were tested in a study of 48 cerebral palsied individuals: 12 anarthric, 12 dysarthric, and 24 controls individually matched to the speech-impaired subjects. There was no impairment of memory span in speech-impaired subjects, who showed normal phonological similarity and word-length effects in short-term memory. Speech-impaired subjects did not differ from their controls in ability to tell whether names of pairs of pictures rhymed. These results challenge the notion that “articulatory coding” is implicated in short-term memory and rhyme judgement and suggests that processes such as rehearsal and phonemic segmentation involve generation of a more abstract central phonological code.
Everyday Reasoning with Conditional Sequences
Authors: Byrne Ruth M. J.
Published: September 2011

Everyday conditional reasoning commonly occurs in a sequence, such as:

… If she leaves home early today then she will get to work quickly. If she gets to work quickly then she will get extra things done …

Experimental studies of conditional reasoning assume that people make the same amount of inferences from sequences as they do from simple arguments that contain just a single conditional premise. In a series of four experiments this assumption is shown to be wrong. Our first experiment showed that people make more inferences from transitive sequences, and fewer inferences from certain atransitive sequences, than from corresponding simple arguments. We established in the second experiment that these differences arise because people can construct a single representation of a sequence. The third experiment replicated the result that people make fewer inferences from certain atransitive sequences. In the final experiment we found that people make more inferences from transitive sequences, regardless of the presence of negatives. We argue that the inferences people make from common everyday sequences depends on their representation of the state of affairs that sequences describe.
Auditory and Visual Word-Stem Completion: Separating Data-Driven and Conceptually Driven Processes
Authors: Bassili John N. , Smith Marilyn C. , MacLeod Colin M.
Published: September 2011

Two experiments investigated the contributions of data-driven and conceptually driven processing on an implicit word-stem completion task. In Experiment 1, individual words were studied either visually or auditorily and were tested using either visual or auditory word-stems. Keeping modality the same from study to test led to more priming than did changing modality, but there was reliable cross-modal priming. In Experiment 2, subjects read sentences like The boat travelled underwater and inferred the subject noun (i.e. “submarine”) or sentences like The submarine travelled underwater and categorized the subject noun (i.e. “boat”). At test, there was reliable priming for both actually read nouns and inferred nouns. In addition, a modality effect was evident for the actually read nouns but not for the inferred nouns. Taken together, these results imply that there is a basic conceptually driven contribution to priming plus an additional contribution of data-driven processing when surface form is the same at study and test.
Manual Asymmetries in Aimed Movements
Authors: Roy Eric A. , Elliott Digby
Published: September 2011

Three hypotheses for the right-hand advantage in aiming movements were examined in these experiments: (1) the right-hand system is more efficient at processing visual information during the movement; (2) subjects make more use of visual information prior to movement initiation when using the right hand; (3) the right hand is less variable in generating force in initiating the pointing response as force demands increase. In the first experiment subjects pointed at a target located directly in front of them from two starting positions which defined short (25-cm) and long (35-cm) movements. The movements were made in three movement times, fast (150 to 249 msec), medium (250 to 349 msec) and slow (350 to 449 msec), under three vision conditions—full vision, and no vision (lights out) with immediate or delayed movement initiation. Performance was measured in movement time and accuracy in amplitude of movement. The results did not completely support any of the hypotheses regarding the right-hand advantage, although the left hand was generally more variable than the right. Also, variability increased with increases in movement length and decreases in movement time. The second experiment was designed to examine further the hypotheses regarding the right-hand advantage. In this experiment the same three visual conditions were used; however, subjects made only fast (<250-msec) movements. Also six rather than two starting positions were used. The increased variability of the left hand was observed again here. Further pointing accuracy with the left hand was more adversely affected in the no-vision delay condition. The implications of these results were discussed as they pertain to understanding the processes involved in visual aiming and the observed manual asymmetries.
Insight without Awareness: On the Interaction of Verbalization, Instruction and Practice in a Simulated Process Control Task
Authors: Stanley William B. , Mathews Robert C. , Buss Ray R. , Kotler-Cope Susan
Published: September 2011

Four experiments in which subjects learned to control two versions of a complex simulated process control task show that verbalizable knowledge of procedures used to perform these tasks is very limited and is acquired late in learning. Individual learning curves associated with these tasks showed sudden improvements in performance, which were not accompanied by a similar increase in verbalizable knowledge. It was also found that verbal instructions consisting of exemplar memorization, strategies for rule induction, simple heuristics, and experts’ instructions were all effective in enhancing novice subjects’ performance. A theoretical framework is proposed in which subjects draw on two separate but interacting knowledge structures to perform these tasks. One knowledge structure is based on memory for past experiences (close analogies), and the other is based on one's current mental model of the task. Implicit sets of competing rules that control response selection are derived from both sources of knowledge. It is suggested that dissociations between task performance and verbalizing occur because memory-based processing tends to have more control over response selection because of its greater specificity, whereas a mental model tends to be the preferred mode for verbal reporting because of its greater accessibility.
Attention Effects on Form Discrimination at Different Eccentricities
Authors: Cheal MaryLou , Lyon Don
Published: September 2011

Considerable disagreement exists in the visual attention literature about how attention is allocated over the visual field. One frequently expressed metaphor is that attention moves like a spotlight, and in some variants it is assumed that attention takes longer to shift to targets further from fixation. In order to test this metaphor, five experiments were conducted in which target location was precued and form discrimination accuracy was assessed. By varying the interval between the precue and the target (stimulus onset asynchrony, SOA), a time course of attention effects was obtained for targets at 2°, 6°, and 10° eccentricity. In the first three experiments, precueing effects were found, but there were no differences in performance as a function of eccentricity for very short SOAs, with either a peripheral cue or a foveal arrow cue. For long SOAs, however, performance was better for targets that were closer to fixation. In Experiments 4 (peripheral cue) and 5 (foveal cue), the targets were scaled to make them equally discriminable at all eccentricities. Again precueing effects were found, but there were no differences in accuracy as a function of eccentricity for most SOAs. These results suggest that attention shifting is not analogous to a constant-velocity moving spotlight.
Spatial Cueing and the Relation between the Accuracy of “Where” and “What” Decisions in Visual Search
Authors: Müller Hermann J. , Rabbitt Patrick M. A.
Published: September 2011

To study the processes underlying selective attention in visual search, the relation between the accuracy of “where” (location) and “what” (same/different orientation matching) decisions was analysed under various display conditions. Target-non-target discriminability was varied by contrasting single and multiple element displays; further, attention was directly manipulated by spatial cueing. In Experiment 1, analyses for both single and multiple displays showed that localization accuracy remained above chance when same/different matching failed; the inverse also obtained. It seems that accurate matching is not a prerequisite for target localization, nor is accurate localization a prerequisite for same/different matching. However, localization is a prerequisite for the accurate recognition of target orientation (Experiment 2). In this case, it seems that features critical for localization “call” attention to a particular candidate location. This facilitates further (shape) analysis of the stimulus that is found there. This orienting process is by-passed if attention is cued to the location in advance.
Tyrosine phosphorylation within the SH3 domain regulates CAS subcellular localization, cell migration, and invasiveness
Authors: Janoštiak Radoslav , Tolde Ondřej , Brůhová Zuzana , Novotný Marian , Hanks Steven K. , Rösel Daniel , Brábek Jan
Published: September 2011

Crk-associated substrate (CAS) is a major tyrosine-phosphorylated protein in cells transformed by v-crk and v-src oncogenes and plays an important role in invasiveness of Src-transformed cells. A novel phosphorylation site on CAS, Tyr-12 (Y12) within the ligand-binding hydrophobic pocket of the CAS SH3 domain, was identified and found to be enriched in Src-transformed cells and invasive human carcinoma cells. To study the biological significance of CAS Y12 phosphorylation, phosphomimicking Y12E and nonphosphorylatable Y12F mutants of CAS were studied. The phosphomimicking mutation decreased interaction of the CAS SH3 domain with focal adhesion kinase (FAK) and PTP-PEST and reduced tyrosine phosphorylation of FAK. Live-cell imaging showed that green fluorescent protein–tagged CAS Y12E mutant is, in contrast to wild-type or Y12F CAS, excluded from focal adhesions but retains its localization to podosome-type adhesions. Expression of CAS-Y12F in cas–/– mouse embryonic fibroblasts resulted in hyperphosphorylation of the CAS substrate domain, and this was associated with slower turnover of focal adhesions and decreased cell migration. Moreover, expression of CAS Y12F in Src-transformed cells greatly decreased invasiveness when compared to wild-type CAS expression. These findings reveal an important role of CAS Y12 phosphorylation in the regulation of focal adhesion assembly, cell migration, and invasiveness of Src-transformed cells.
Diacylglycerol kinase ζ controls diacylglycerol metabolism at the immunological synapse
Authors: Gharbi Severine I. , Rincón Esther , Avila-Flores Antonia , Torres-Ayuso Pedro , Almena María , Cobos María Angeles , Albar Juan Pablo , Mérida Isabel
Published: September 2011

Diacylglycerol (DAG) generation at the T cell immunological synapse (IS) determines the correct activation of antigen-specific immune responses. DAG kinases (DGKs) α and ζ act as negative regulators of DAG-mediated signals by catalyzing DAG conversion to phosphatidic acid (PA). Nonetheless, the specific input of each enzyme and their spatial regulation during IS formation remain uncharacterized. Here we report recruitment of endogenous DGKα and DGKζ to the T cell receptor (TCR) complex following TCR/CD28 engagement. Specific DGK gene silencing shows that PA production at the activated complex depends mainly on DGKζ, indicating functional differences between these proteins. DGKζ kinase activity at the TCR is enhanced by phorbol-12-myristate-13-acetate cotreatment, suggesting DAG-mediated regulation of DGKζ responsiveness. We used GFP-DGKζ and -DGKα chimeras to assess translocation dynamics during IS formation. Only GFP-DGKζ translocated rapidly to the plasma membrane at early stages of IS formation, independent of enzyme activity. Finally, use of a fluorescent DAG sensor confirmed rapid, sustained DAG accumulation at the IS and allowed us to directly correlate membrane translocation of active DGKζ with DAG consumption at the IS. This study highlights a DGKζ-specific function for local DAG metabolism at the IS and offers new clues to its mode of regulation.
α-Catenin contributes to the strength of E-cadherin–p120 interactions
Authors: Troyanovsky Regina B. , Klingelhöfer Jörg , Troyanovsky Sergey M.
Published: September 2011

Cadherin–catenin interactions play an important role in cadherin-mediated adhesion. Here we present strong evidence that in the cadherin–catenin complex α-catenin contributes to the binding strength of another catenin, p120, to the same complex. Specifically, we found that a β-catenin–uncoupled cadherin mutant interacts much more weakly with p120 than its full-size counterpart and that it is rapidly endocytosed from the surface of A-431 cells. We also showed that p120 overexpression stabilizes this mutant on the cell surface. Examination of the α-catenin–deficient MDA-MB-468 cells and their derivates in which α-catenin was reintroduced showed that α-catenin reinforces E-cadherin–p120 association. Finally, a cross-linking analysis of the cadherin–catenin complex indicated that a large loop located in the middle of the p120 arm-repeat domain is in close spatial vicinity to the amino-terminal VH1 domain of α-catenin. The six amino acid–long extension of this loop, caused by an alternative splicing, weakens p120 binding to cadherin. The data suggest that α-catenin–p120 contact within the cadherin–catenin complex can regulate cadherin trafficking.
Nesprin-3 regulates endothelial cell morphology, perinuclear cytoskeletal architecture, and flow-induced polarization
Authors: Morgan Joshua T. , Pfeiffer Emily R. , Thirkill Twanda L. , Kumar Priyadarsini , Peng Gordon , Fridolfsson Heidi N. , Douglas Gordon C. , Starr Daniel A. , Barakat Abdul I.
Published: September 2011

Changes in blood flow regulate gene expression and protein synthesis in vascular endothelial cells, and this regulation is involved in the development of atherosclerosis. How mechanical stimuli are transmitted from the endothelial luminal surface to the nucleus is incompletely understood. The linker of nucleus and cytoskeleton (LINC) complexes have been proposed as part of a continuous physical link between the plasma membrane and subnuclear structures. LINC proteins nesprin-1, -2, and -4 have been shown to mediate nuclear positioning via microtubule motors and actin. Although nesprin-3 connects intermediate filaments to the nucleus, no functional consequences of nesprin-3 mutations on cellular processes have been described. Here we show that nesprin-3 is robustly expressed in human aortic endothelial cells (HAECs) and localizes to the nuclear envelope. Nesprin-3 regulates HAEC morpho­logy, with nesprin-3 knockdown inducing prominent cellular elongation. Nesprin-3 also organizes perinuclear cytoskeletal organization and is required to attach the centrosome to the nuclear envelope. Finally, nesprin-3 is required for flow-induced polarization of the centrosome and flow-induced migration in HAECs. These results represent the most complete description to date of nesprin-3 function and suggest that nesprin-3 regulates vascular endothelial cell shape, perinuclear cytoskeletal architecture, and important aspects of flow-mediated mechanotransduction.
Tool Path Planning for Near-Dry EDM Milling With Lead Angle on Curved Surfaces
Authors: Fujiki Masahiro , Ni Jun , Shih Albert J.
Published: September 2011

This research investigates the strategy to achieve high material removal rate in tool path planning for the near-dry electrical discharge machining (EDM) milling process using tubular electrode with a lead angle. The proposed strategy to prevent leakage of dielectric mist from the tubular electrode is different from the conventional end milling process due to the difference in material removal mechanism. Tool positions and orientations to engage the electrode into workpiece, machining of workpiece edge, minimum lead angle to machine a curved surface, and minimum and maximum path interval to prevent the mist leakage are derived. Experiments are conducted to validate the model prediction of path planning. Experimental results show plunge method has the highest material removal rate for engaging method, and electrode hole must be located within the workpiece surface when edge of workpiece is machined. For curvature machining, the proposed path planning strategy yields higher material removal rate compared with that from the conventional strategy, which only avoids gouging. This study also reveals that, due to the tool wear and crowning of electrode tip, it is difficult to accurately determine the minimum path interval which will cause the mist leakage.
Analysis of Generation Capabilities of Noncircular Cogbelt Pulleys on the Example of a Gear With an Elliptical Pitch Line
Authors: Kujawski Mieczysław , Krawiec Piotr
Published: September 2011

The aim of this paper is the analysis of the generation capabilities of noncircular cogbelt pulleys on the example of a cogbelt pulley with an elliptical pitch line. The need of elaboration of different manufacturing methods for noncircular cogbelt pulleys than existing ones for noncircular gears is shown. Relationships for the mutual position of the cutting tool and noncircular cogbelt pulley during teeth generating are given. The profile of the tool’s cutting edge, which provides the required shape of tooth spaces, is elaborated. Recommendations for designers and process engineers, who deal with elaboration of technology for noncircular cogbelt pulleys, are formulated.
Megacity Emissions and Lifetimes of Nitrogen Oxides Probed from Space
Authors: Beirle Steffen , Boersma K. Folkert , Platt Ulrich , Lawrence Mark G. , Wagner Thomas
Published: September 2011

Analysis of downwind plume evolution using satellite observations can be used for air pollution estimates.
Field Work as Art and Science

Field Notes on Science and Nature
Michael R. Canfield, Ed.

Harvard University Press, Cambridge, MA, 2011. 313 pp. $27.95, £20.95, €25.20. ISBN 9780674057579.
Authors: Greene Harry W.
Published: September 2011

Through essays that incorporate examples from their own notebooks, the contributors describe their experiences in and methods for recording field observations.
Antibiotic Resistance, Not Shaken or Stirred
Authors: Frisch Ryan L. , Rosenberg Susan M.
Published: September 2011

Microdevices for microbial culture may better capture the evolution of antibiotic resistance in real-world environments.
Coexisting with Cattle
Authors: du Toit Johan T.
Published: September 2011

In East Africa, large wild herbivores both compete with and benefit cattle.
Biodiversity and Productivity
Authors: Willig Michael R.
Published: September 2011

The relationship between species richness and ecosystem productivity appears to be very complex.
Webb Telescope Pulled Back From the Brink—For Now
Authors: Bhattacharjee Yudhijit
Published: September 2011

Last week, a Senate panel threw a lifeline to the James Webb Space Telescope by allocating $530 million for the project, $156 million more than the Administration's request.
Tracing the Paths of the First Americans
Authors: Balter Michael
Published: September 2011

New findings support earlier indications that the Paleoindians, the ancestors of today's Native Americans, stem from a single Asian source population.
Immunogenicity of Recombinant Classic Swine Fever Virus CD8+T Lymphocyte Epitope and Porcine Parvovirus VP2 Antigen Coexpressed by Lactobacillus casei in Swine via Oral Vaccination
Authors: Xu Yigang , Cui Lichun , Tian Changyong , Zhang Guocai , Huo Guicheng , Tang Lijie , Li Yijing
Published: September 2011

Classical swine fever virus (CSFV) and porcine parvovirus (PPV) are highly contagious pathogens, resulting in enormous economic losses in pig industries worldwide. Because vaccines play an important role in disease control, researchers are seeking improved vaccines that could induce antiviral immune responses against CSFV and PPV at the mucosal and systemic levels simultaneously. In this study, a genetically engineeredLactobacillusstrain coexpressing the CSFV-specific cytotoxic T lymphocyte (CTL) epitope 290 and the VP2 antigen of PPV was developed, and its immunopotentiating capacity as an oral vaccine in pigs was analyzed. The data demonstrated that in the absence of any adjuvant, the recombinantLactobacillusstrain can efficiently stimulate mucosal and systemic CSFV-specific CD8+CTL responses to protect pigs against CSFV challenge. Moreover, anti-PPV-VP2 serum IgG and mucosal IgA were induced in pigs immunized orally with the recombinantLactobacillusstrain, showing a neutralizing effect on PPV infection. The results suggest that the recombinantLactobacillusmicroecological agent may be a valuable component of a strategy for development of a vaccine against CSFV and PPV.
SmaI Restriction Site-Based Multiplex PCR for Typing of Hospital- and Community-Acquired Staphylococcus aureus
Authors: Al-Zahrani Ibrahim A. , Hamson Clare , Edge David , Collins Jennifer , Perry John D. , Raza Muhammad , Gould Kate , Harwood Colin R.
Published: September 2011

Methicillin-resistant
Staphylococcus aureus
(MRSA) is an important nosocomial pathogen, and morbidity and mortality rates associated with this pathogen have increased markedly in recent years. MRSA strains are generally resistant to several classes of antibiotics and are therefore difficult and costly to treat. A major issue is to identify the sources of MRSA infections and to monitor their epidemic spread. In this study, we report the development of a typing technique for
S. aureus
, based on single-nucleotide polymorphism (SNP) variations in and around SmaI-restriction sites (CCCGGG). An assessment of the SmaI restriction site-based multiplex PCR (SmaI-multiplex PCR) typing (SMT) with respect to pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) revealed a high level of concordance in the clustering of the test strains. The SmaI-multiplex PCR was found to be more discriminatory than MLST/staphylococcal cassette chromosome
mec
(SCC
mec
) typing but less discriminatory than PFGE. SMT can provide real-time information for the investigation of ongoing
S. aureus
hospital outbreaks. SMT meets the criteria of a practical typing method: it is simple, reproducible, and highly discriminatory and does not require expensive equipment or specialist expertise. Consequently, SmaI-multiplex PCR has the potential to be used in routine clinical microbiology laboratories.
Target-Specific Capture Enhances Sensitivity of Electrochemical Detection of Bacterial Pathogens
Authors: Patel Mayank , Gonzalez Rodrigo , Halford Colin , Lewinski Michael A. , Landaw Elliot M. , Churchill Bernard M. , Haake David A.
Published: September 2011

We report the concentration and purification of bacterial 16S rRNA by the use of a biotinylated DNA target-specific capture (TSC) probe. For both cultivated bacterial and urine specimens from urinary tract infection patients, TSC resulted in a 5- to 8-fold improvement in the sensitivity of bacterial detection in a 16S rRNA electrochemical sensor assay.
The Cellular TAR RNA Binding Protein, TRBP, Promotes HIV-1 Replication Primarily by Inhibiting the Activation of Double-Stranded RNA-Dependent Kinase PKR
Authors: Sanghvi Viraj R. , Steel Laura F.
Published: September 2011

The TAR RNA binding protein, TRBP, is a cellular double-stranded RNA (dsRNA) binding protein that can promote the replication of HIV-1 through interactions with the viral TAR element as well as with cellular proteins that affect the efficiency of translation of viral transcripts. The structured TAR element, present on all viral transcripts, can impede efficient translation either by sterically blocking access of translation initiation factors to the 5′-cap or by activating the dsRNA-dependent kinase, PKR. Several mechanisms by which TRBP can facilitate translation of viral transcripts have been proposed, including the binding and unwinding of TAR and the suppression of PKR activation. Further, TRBP has been identified as a cofactor of Dicer in the processing of microRNAs (miRNAs), and sequestration of TRBP by TAR in infected cells has been proposed as a viral countermeasure to potential host cell RNA interference-based antiviral activities. Here, we have addressed the relative importance of these various roles for TRBP in HIV-1 replication. Using Jurkat T cells, primary human CD4
+
T cells, and additional cultured cell lines, we show that depletion of TRBP has no effect on viral replication when PKR activation is otherwise blocked. Moreover, the presence of TAR-containing mRNAs does not affect the efficacy of cellular miRNA silencing pathways. These results establish that TRBP, when expressed at physiological levels, promotes HIV-1 replication mainly by suppressing the PKR-mediated antiviral response, while its contribution to HIV-1 replication through PKR-independent pathways is minimal.
A Sequence within the Varicella-Zoster Virus (VZV) OriS Is a Negative Regulator of DNA Replication and Is Bound by a Protein Complex Containing the VZV ORF29 Protein
Authors: Khalil Mohamed I. , Arvin Ann , Jones Jeremy , Ruyechan William T.
Published: September 2011

The architecture of the varicella-zoster virus (VZV) origin of DNA replication (OriS) differs significantly from that of the herpes simplex virus (HSV) DNA replication origin. Novel aspects of the VZV OriS include a GA-rich region, three binding sites for the VZV origin-binding protein (OBP) all on the same strand and oriented in the same direction, and a partial OBP binding site of unknown function. We have designated this partial binding site Box D and have investigated the role it plays in DNA replication and flanking gene expression. This has been done with a model system using a replication-competent plasmid containing OriS and a replication- and transcription-competent dual-luciferase reporter plasmid containing both the OriS and the intergenic region between VZV open reading frames (ORFs) 62 and 63. We have found that (i) Box D is a negative regulator of DNA replication independent of flanking gene expression, (ii) the mutation of Box D results in a decrease in flanking gene expression, thus a sequence within the VZV OriS affects transcription, which is in contrast to results reported for HSV-1, (iii) there is a specific Box D complex formed with infected cell extracts in electrophoretic mobility shift assay experiments, (iv) supershift assays show that this complex contains the VZV ORF29 single-strand DNA-binding protein, and (v) the formation of this complex is dependent on the presence of CGC motifs in Box D and its downstream flanking region. These findings show that the VZV ORF29 protein, while required for DNA replication, also plays a novel role in the suppression of that process.
A Novel Borna Disease Virus Vector System That Stably Expresses Foreign Proteins from an Intercistronic Noncoding Region
Authors: Daito Takuji , Fujino Kan , Honda Tomoyuki , Matsumoto Yusuke , Watanabe Yohei , Tomonaga Keizo
Published: September 2011

Borna disease virus (BDV), a nonsegmented, negative-strand RNA virus, infects a wide variety of mammalian species and readily establishes a long-lasting, persistent infection in brain cells. Therefore, this virus could be a promising candidate as a novel RNA virus vector enabling stable gene expression in the central nervous system (CNS). Previous studies demonstrated that the 5′ untranslated region of the genome is the only site for insertion and expression of a foreign gene. In this study, we established a novel BDV vector in which an additional transcription cassette has been inserted into an intercistronic noncoding region between the viral phosphoprotein (P) and matrix (M) genes. The recombinant BDV (rBDV) carrying green fluorescent protein (GFP) between the P and M genes, rBDV P/M-GFP, expressed GFP efficiently in cultured cells and rodent brains for a long period of time without attenuation. Furthermore, we generated a nonpropagating rBDV, ΔGLLP/M, which lacks the envelope glycoprotein (G) and a splicing intron within the polymerase gene (L), by the transcomplementation system with either transient or stable expression of the G gene. Interestingly, rBDV ΔGLLP/M established a persistent infection in cultured cells with stable expression of GFP in the absence of the expression of G. Using persistently infected rBDV ΔGLLP/M-infected cells, we determined the amino acid region in the cytoplasmic tail (CT) of BDV G important for the release of infectious rBDV particles and also demonstrated that the CT region may be critical for the generation of pseudotyped rBDV having vesicular stomatitis virus G protein. Our results revealed that the newly established BDV vector constitutes an alternative tool not only for stable expression of foreign genes in the CNS but also for understanding the mechanism of the release of enveloped virions.
Analysis of Single-Nucleotide Polymorphisms in Patient-Derived Retrovirus Integration Sites Reveals Contamination from Cell Lines Acutely Infected by Xenotropic Murine Leukemia Virus-Related Virus
Authors: Rusmevichientong Alice , Das Gupta Jaydip , Elias Petra S. , Silverman Robert H. , Chow Samson A.
Published: September 2011

We analyzed xenotropic murine leukemia virus-related virus (XMRV) integration site sequences previously identified from human prostate tissues for single-nucleotide polymorphisms (SNPs) to discriminate between patient and potential cell line sources of the proviruses. The SNPs of two integration sites were identical to those in cell lines but not the patients, whereas the data on the remaining 12 integration sites were inconclusive. Our results provide direct evidence for contamination during analysis of XMRV integration sites.
Resveratrol-Mediated Gamma Interferon Reduction Prevents Airway Inflammation and Airway Hyperresponsiveness in Respiratory Syncytial Virus-Infected Immunocompromised Mice
Authors: Zang Na , Xie Xiaohong , Deng Yu , Wu Shengde , Wang Lijia , Peng Caijing , Li Simin , Ni Ke , Luo Yan , Liu Enmei
Published: September 2011

Respiratory syncytial virus (RSV) is the most important cause of severe, lower respiratory tract infections in infants, and RSV infections have been associated with chronic wheezing and asthma during childhood. However, the mechanism of RSV-induced airway inflammation and airway hyperresponsiveness (AHR) is poorly understood. Furthermore, there are presently neither effective vaccines nor drugs available for the prevention or treatment of RSV infections. In this study, we investigated the effect of the plant extract resveratrol as a means of preventing airway inflammation and attenuating RSV-induced AHR. Our data showed that resveratrol reduced RSV lung titers and the number of infiltrating lymphocytes present in bronchoalveolar lavage fluid (BALF) and reduced inflammation. Furthermore, resveratrol attenuated airway responses to methacholine following RSV infection and significantly decreased gamma interferon (IFN-γ) levels in BALF of RSV-infected mice. Data presented in this report demonstrated that resveratrol controlled Toll-like receptor 3 (TLR3) expression, inhibited the TRIF signaling pathway, and induced M2 receptor expression following RSV infection. These data support a role for the use of resveratrol as a means of reducing IFN-γ levels associated with RSV-mediated airway inflammation and AHR, which may be mediated via TLR3 signaling.
Modeling of the ribonucleotide reductases substrate reaction. Hydrogen atom abstraction by a thiyl free radical and detection of the ribosyl-based carbon radical by pulse radiolysis
Authors: Wnuk Stanislaw F. , Penjarla Jaidev A. K. , Dang Thao , Mebel Alexander M. , Nauser Thomas , Schöneich Christian
Published: September 2011

The 1,4-anhydro-5-deoxy-6-thio-D-ribo-hexofuranitol (1) was prepared from 1,2-O-isopropylidene-α-D-glucose in 10 steps. In a key step treatment of the 1,2-O-isopropylidenehexofuranose derivative with BF3/Et3SiH effected deacetonization and reductive deoxygenation at carbon 1. Pulse radiolysis experiments with 6-thiohexofuranitol 1 and its disulfide derivative demonstrated formation of the ribosyl-based carbon-centered radical upon generation of 6-thiyl radical in basic medium. The proposed [1,5]-hydrogen shift abstraction with generation of the C3 radical mimics the initial substrate reaction of RNRs. The reversible H-atom transfer has been quantified and was correlated with the computed rate constants for the internal H atom abstraction from C1, C2, C3 and C4 by the thiyl radical. The energy barrier for the H3 and H4 abstractions were calculated to be most favorable with the corresponding barriers of 11.1 and 11.2 kcal/mol, respectively.
Acute and chronic effects of SGLT2 blockade on glomerular and tubular function in the early diabetic rat
Authors: Thomson Scott C. , Rieg Timo , Miracle Cynthia , Mansoury Hadi , Whaley Jean , Vallon Volker , Singh Prabhleen
Published: September 2011

Tubuloglomerular feedback (TGF) stabilizes nephron function from minute to minute and adapts to different steady-state inputs to maintain this capability. Such adaptation inherently renders TGF less efficient at buffering long-term disturbances, but the magnitude of loss is unknown. We undertook the present study to measure the compromise between TGF and TGF adaptation in transition from acute to chronic decline in proximal reabsorption (Jprox). As a tool, we blocked proximal tubule sodium-glucose cotransport with the SGLT2 blocker dapagliflozin in hyperglycemic rats with early streptozotocin diabetes, a condition in which a large fraction of proximal fluid reabsorption owes to SGLT2. Dapagliflozin acutely reduced proximal reabsorption leading to a 70% increase in early distal chloride, a saturated TGF response, and a major reduction in single nephron glomerular filtration rate (SNGFR). Acute and chronic effects on Jprox were indistinguishable. Adaptations to 10–12 days of dapagiflozin included increased reabsorption by Henle's loop, which caused a partial relaxation in the increased tone exerted by TGF that could be explained without desensitization of TGF. In summary, TGF contributes to long-term fluid and salt balance by mediating a persistent decline in SNGFR as the kidney adapts to a sustained decrease in Jprox.
Large BP-dependent and -independent differences in susceptibility to nephropathy after nitric oxide inhibition in Sprague-Dawley rats from two major suppliers
Authors: Griffin Karen , Polichnowski Aaron , Licea-Vargas Hector , Picken Maria , Long Jianrui , Williamson Geoffrey , Bidani Anil
Published: September 2011

The Nω-nitro-l-arginine methyl ester (l-NAME) model is widely employed to investigate the role of nitric oxide (NO) in renal injury. The present studies show that Sprague-Dawley rats from Harlan (H) and Charles River (CR) exhibit strikingly large differences in susceptibility to l-NAME nephropathy. After 4 wk of l-NAME (∼50 mg·kg−1·day−1in drinking water), H rats ( n = 13) exhibited the expected hypertension [average radiotelemetric systolic blood pressure (BP), 180 ± 3 mmHg], proteinuria (136 ± 17 mg/24 h), and glomerular injury (GI) (12 ± 2%). By contrast, CR rats developed less hypertension (142 ± 4), but surprisingly no proteinuria or GI, indicating a lack of glomerular hypertension. Additional studies showed that conscious H, but not CR, rats exhibit dose-dependent renal vasoconstriction after l-NAME. To further investigate these susceptibility differences, l-NAME was given 2 wk after 3/4 normotensive nephrectomy (NX) and comparably impaired renal autoregulation in CR-NX and H-NX rats. CR-NX rats, nevertheless, still failed to develop proteinuria and GI despite moderate hypertension (144 ± 2 mmHg, n = 29). By contrast, despite an 80–90% l-NAME dose reduction and lesser BP increases (169 ± 4 mmHg), H-NX rats ( n = 20) developed greater GI (26 ± 3%) compared with intact H rats. Linear regression analysis showed significant ( P < 0.01) differences in the slope of the relationship between BP and GI between H-NX (slope 0.56 ± 0.14; r = 0.69; P < 0.008) and CR-NX (slope 0.09 ± 0.06; r = 0.29; P = 0.12) rats. These data indicate that blunted BP responses to l-NAME in the CR rats are associated with BP-independent resistance to nephropathy, possibly mediated by a resistance to the renal (efferent arteriolar) vasoconstrictive effects of NO inhibition.
Use of mouse hematopoietic stem and progenitor cells to treat acute kidney injury
Authors: Li Ling , Black Rachel , Ma Zhendong , Yang Qiwen , Wang Andrew , Lin Fangming
Published: September 2011

New and effective treatment for acute kidney injury remains a challenge. Here, we induced mouse hematopoietic stem and progenitor cells (HSPC) to differentiate into cells that partially resemble a renal cell phenotype and tested their therapeutic potential. We sequentially treated HSPC with a combination of protein factors for 1 wk to generate a large number of cells that expressed renal developmentally regulated genes and protein. Cell fate conversion was associated with increased histone acetylation on promoters of renal-related genes. Further treatment of the cells with a histone deacetylase inhibitor improved the efficiency of cell conversion by sixfold. Treated cells formed tubular structures in three-dimensional cultures and were integrated into tubules of embryonic kidney organ cultures. When injected under the renal capsule, they integrated into renal tubules of postischemic kidneys and expressed the epithelial marker E-cadherin. No teratoma formation was detected 2 and 6 mo after cell injection, supporting the safety of using these cells. Furthermore, intravenous injection of the cells into mice with renal ischemic injury improved kidney function and morphology by increasing endogenous renal repair and decreasing tubular cell death. The cells produced biologically effective concentrations of renotrophic factors including VEGF, IGF-1, and HGF to stimulate epithelial proliferation and tubular repair. Our study indicates that hematopoietic stem and progenitor cells can be converted to a large number of renal-like cells within a short period for potential treatment of acute kidney injury.
Analyses of the facilitatory effect of orexin on eating and masticatory muscle activity in rats
Authors: Tsuji Tadataka , Yamamoto Takashi , Tanaka Susumu , Bakhshishayan Sanam , Kogo Mikihiko
Published: September 2011

The orexins (orexin-A and orexin-B) are neuropeptides that are secreted from neurons in the lateral hypothalamus and that participate in the regulation of feeding behavior. It remains to be determined, however, how the orexins exert their effects on feeding behavior, including masticatory movements. To this end, we analyzed food intake behavior and masticatory muscle activity using video analysis and electromyography (EMG) recording methods. The results showed that the cumulative food intake over 4 h was larger in rats intraventricularly injected with either orexin-A or orexin-B than in saline-injected control rats. The latency to eating and the feeding time for a fixed amount of pellets were shortened by injections of orexins in a dose-dependent manner, with a more potent effect by orexin-A than orexin-B. The shorter feeding time corresponded to a decreased number of chewing cycles. EMG recordings from both the digastric and masseter muscles showed two distinct patterns of bursts corresponding to the gnawing and chewing phases. After the injection of orexin-A, the magnitude of the bursts became larger in both phases in the masseter muscle, the burst duration became longer in the chewing phase in the masseter muscle, and the interburst interval became shortened in the gnawing phase in both muscles. Consequently, the burst frequency in the chewing phase was increased in the digastric muscle and, conversely, reduced in the masseter muscle. These results suggest that the orexin-A-induced facilitatory feeding behavior is characterized by a dynamic jaw-opener activity that opens the mouth rapidly and a powerful jaw-closer activity for crushing the increased amount of food taken into the mouth. The possible involvement of orexin-A in binge eating disorder is discussed.
Comments on the Extraperitoneal Approach for Standard Laparoscopic Radical Prostatectomy: What Is Gained and What Is Lost
Authors: Liatsikos Evangelos , Kyriazis Iason , Kallidonis Panagiotis , Do Minh , Haefner Tim , Dietel Anja , Holze Sigrun , Ragavan Narasimhan , Stolzenburg Jens-Uwe
Published: September 2011

Laparoscopic extraperitoneal radical prostatectomy (LERP) is considered the standard care treatment option for the management of localized and locally advanced prostatic cancer (PCa) in many institutes worldwide. In this work, the main advantages and disadvantages of LERP approach are reviewed with regard to its outcomes, the complication management, the learning curve, and the extend of pelvic lymph node dissection (PLND). It is concluded that LERP demonstrates comparable cancer control, urinary continence, and potency outcomes with the open and the robot-assisted radical prostatectomy, while offering advantages in complication management in comparison to the transperitoneal approach. Learning curve of LERP is considered long and stiff and significantly affects perioperative outcomes and morbidity, cancer control, and functional results. Thus, close mentoring especially in the beginning of the learning curve is advised. Finally, LERP still has a role in the limited or modified PLND offered in intermediate risk PCa patients.
Visceral Abdominal and Subfascial Femoral Adipose Tissue Have Opposite Associations with Liver Fat in Overweight and Obese Premenopausal Caucasian Women
Authors: Rocha Paulo M. , Barata José T. , Minderico Cláudia S. , Silva Analiza M. , Teixeira Pedro J. , Sardinha Luís B.
Published: September 2011

Abdominal obesity has been associated with liver fat storage. However, the relationships between other body composition depots and metabolic syndrome features with hepatic fat are still unclear. We examined abdominal and thigh adipose tissue (AT) compartments associations with liver fat in 140 overweight and obese premenopausal Caucasian women. Blood lipids and, proinflammatory and atherothrombotic markers associations with hepatic fat were also analyzed. A larger visceral AT (VAT) was related with liver fat (P<0.05). Contrarily, thigh subfascial AT was inversely related to liver fat (P<0.05). Increased fasting insulin, triglycerides, PAI-1 concentrations, and a higher total-cholesterol/HDL-cholesterol ratio were also associated with hepatic fat, even after adjustment for VAT (P<0.05). Thigh subfascial adiposity was inversely associated with liver fat, suggesting a potential preventive role against ectopic fat storage in overweight and obese women. These results reinforce the contribution of an abdominal obesity phenotype associated with a diabetogenic and atherothrombotic profile to liver lipotoxicity.