bluuebunny/crossref_metadata_2025 huggingface dataset:
Propofol and Thiopental Suppress Amyloid Fibril Formation and GM1 Ganglioside Expression through the γ-Aminobutyric Acid A Receptor
Authors: Yamamoto Naoki , Arima Hajime , Sugiura Takeshi , Hirate Hiroyuki , Taniura Hideo , Suzuki Kenji , Sobue Kazuya
Published: February 2013

Background:

The incidence of Alzheimer disease may increase after surgical interventions. Amyloid β-protein (Aβ) fibrillogenesis, which is closely related to Alzheimer disease, is reportedly accelerated by exposure to anesthetics. However, the effects of GM1 ganglioside (GM1) on Αβ fibrillogenesis have not yet been reported. The current study was designed to examine whether the anesthetics propofol and thiopental are associated with Αβ assembly and GM1 expression on the neuronal cell surface.

Methods:

PC12N cells and cultured neuronal cells were treated with propofol or thiopental, and GM1 expression in treated and untreated cells was determined by the specific binding of horseradish peroxidase-conjugated cholera toxin subunit B (n = 5). The effects of an inhibitor of the γ-aminobutyric acid A receptor was also examined (n= 5). In addition, the effects of the anesthetics on GM1 liposome-induced Αβ assembly were investigated (n = 5). Finally, the neurotoxicity of the assembled Αβ fibrils was studied by the lactate dehydrogenase release assay (n = 6).

Results:

Propofol (31.2±4.7%) and thiopental (34.6±10.5%) decreased GM1 expression on the cell surface through the γ-aminobutyric acid A receptor. The anesthetics inhibited Αβ fibril formation from soluble Αβ in cultured neurons. Moreover, propofol and thiopental suppressed GM1-induced fibril formation in a cell-free system (propofol, 75.8±1.9%; thiopental, 83.6±1.9%) and reduced the neurotoxicity of a mixture containing Aβ and GM1 liposomes (propofol, 35.3±16.4%; thiopental, 21.3±11.6%).

Conclusions:

Propofol and thiopental have direct and indirect inhibitory effects on Αβ fibrillogenesis.
An integrative, translational approach to understanding rare and orphan genetically based diseases
Authors: Hoehndorf Robert , Schofield Paul N. , Gkoutos Georgios V.
Published: February 2013

PhenomeNet is an approach for integrating phenotypes across species and identifying candidate genes for genetic diseases based on the similarity between a disease and animal model phenotypes. In contrast to ‘guilt-by-association’ approaches, PhenomeNet relies exclusively on the comparison of phenotypes to suggest candidate genes, and can, therefore, be applied to study the molecular basis of rare and orphan diseases for which the molecular basis is unknown. In addition to disease phenotypes from the Online Mendelian Inheritance in Man (OMIM) database, we have now integrated the clinical signs from Orphanet into PhenomeNet. We demonstrate that our approach can efficiently identify known candidate genes for genetic diseases in Orphanet and OMIM. Furthermore, we find evidence that mutations in the
HIP1
gene might cause Bassoe syndrome, a rare disorder with unknown genetic aetiology. Our results demonstrate that integration and computational analysis of human disease and animal model phenotypes using PhenomeNet has the potential to reveal novel insights into the pathobiology underlying genetic diseases.
Devosia submarina sp. nov., isolated from deep-sea surface sediments
Authors: Romanenko Lyudmila A. , Tanaka Naoto , Svetashev Vassilii I.
Published: February 2013

The taxonomic status of two aerobic, Gram-stain-negative, orange–reddish pigmented, motile, rod-shaped bacteria, designated KMM 9415Tand KMM 9416, isolated from a deep surface-sediment sample from the Sea of Japan, was defined. Comparative 16S rRNA gene sequence analysis of strains KMM 9415Tand KMM 9416 revealed their affiliation to the genusDevosiawith a high sequence similarity of 98.5 % to bothDevosia psychrophilaDSM 22950TandDevosia glacialisLMG 26051T. The novel strains were characterized by the predominance of the fatty acid C18 : 1ω7cfollowed by C16 : 1and C16 : 0. The major isoprenoid quinone was Q-10 and the polar lipids comprised phosphatidylglycerol, phosphatidic acid and unknown glycolipids. The DNA–DNA hybridization value of 88 % between the novel strains KMM 9415Tand KMM 9416 confirmed their assignment to the same species. The values of DNA relatedness determined for strain KMM 9415Tand the closely related strainsD. psychrophilaDSM 22950TandD. glacialisLMG 26051Twere 21 % and 23 %, respectively. Based on distinctive phenotypic characteristics, phylogenetic analysis and DNA–DNA relatedness, it can be concluded that the novel strains KMM 9415Tand KMM 9416 represent a novel species within the genusDevosia, for which the nameDevosiasubmarinasp. nov. is proposed. The type strain is the strain KMM 9415T( = NRIC 0884T = JCM 18935T).
Pustulibacterium marinum gen. nov., sp. nov., a member of the family

Flavobacteriaceae

isolated from the Bashi Channel
Authors: Wang Guanghua , Zhou Danyan , Dai Shikun , Tian Xinpeng , Li Jie , Chen Wen , Xiang Wenzhou , Li Xiang
Published: February 2013

A Gram-reaction-negative, non-spore-forming, gliding, non-translucent, colourless or yellow, aerobic and elevated-colony-forming strain, designated E403T, was isolated from the Bashi Channel and subjected to a polyphasic taxonomic study. Strain E403T could grow in the presence of 0.3–8 % (w/v) NaCl, at 16–43 °C and at pH 6–9, and grew optimally at 28 °C, pH 8, in natural seawater medium. The respiratory quinones were MK-6 and MK-7. The major fatty acids were iso-C15 : 0, iso-C17 : 0 3-OH, iso-C15 : 1 G, summed feature 3 (C16 : 1ω7c/C16 : 1ω6c), iso-C15 : 0 3-OH and C16 : 0. The DNA G+C content of strain E403T was 37.9 mol%. Phylogenetic analyses based on 16S rRNA gene sequences of members of the family

Flavobacteriaceae

showed that strain E403T formed a distinct evolutionary lineage within the stable cluster containing type strains

Zhouia amylolytica

HN-171T (92.2 % similarity) and

Joostella marina

En5T (92.4 % similarity). In addition to the large 16S rRNA gene sequence differences, E403T can also be distinguished from the reference type strains

J. marina

En5T and

Sinomicrobium oceani

SCSIO 03483T by several phenotypic characteristics and chemotaxonomic properties. On the basis of phenotypic, chemotaxonomic and phylogenetic properties, strain E403T is suggested to represent a novel species of a new genus in the family

Flavobacteriaceae

, for which the name Pustulibacterium marinum gen. nov., sp. nov. is proposed. The type strain is E403T ( = CCTCC AB2012862T = CGMCC 1.12333T = KCTC 32192T).
Brevifollis gellanilyticus gen. nov., sp. nov., a gellan-gum-degrading bacterium of the phylum

Verrucomicrobia
Authors: Otsuka Shigeto , Suenaga Taku , Vu Hoan Thi , Ueda Hiroyuki , Yokota Akira , Senoo Keishi
Published: February 2013

The taxonomic properties of strain DC2c-G4T, a Gram-staining-negative, ovoid, gellan-gum-degrading bacterial isolate, were examined. Phylogenetic analysis based on 16S rRNA gene sequences identified this isolate as a member of the phylum

Verrucomicrobia

and closest to the genus

Prosthecobacter

. The 16S rRNA gene sequence similarities between this isolate and any of the type strains of species of the genus

Prosthecobacter

were less than 95 %. In addition, the absence of a single prostheca and the predominant menaquinone MK-7(H2) supported the differentiation of this isolate from the genus

Prosthecobacter

. Here, we propose Brevifollis gellanilyticus gen. nov., sp. nov. to accommodate the isolate. The type strain of the type species is DC2c-G4T ( = NBRC 108608T = CIP 110457T).
Rhodospirillum

oryzae sp. nov., a phototrophic bacterium isolated from rhizosphere soil of paddy
Authors: Lakshmi K. V. N. S. , Sasikala Ch. , Ramaprasad E. V. V. , Ramana Ch. V.
Published: February 2013

A reddish-brown bacterium, designated strain JA318T, was purified from a photoheterotrophic enrichment culture obtained from the rhizosphere soil of paddy. Cells of strain JA318T are spiral shaped, Gram-stain-negative and motile by means of amphitrichous flagella. Strain JA318T has no NaCl requirement for growth but can tolerate up to 1.5 % (w/v) NaCl. Internal photosynthetic membranes are present as lamellar stacks. Photoorganoheterotrophy is the only growth mode observed. Strain JA318T contains bacteriochlorophyll a, lycopene and rhodopin as major carotenoids. Thiamine, niacin and para-aminobenzoic acid (PABA) are required as growth factors. Major fatty acids are C18 : 1ω7c and C16 : 0. Ubiquinone-8 and rhodoquinone-8 are the observed quinones. Diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and an unidentified aminolipid are the major polar lipids in strain JA318T. Phylogenetic analysis based on 16S rRNA gene sequences showed that the strain JA318T clustered with species of the genus

Rhodospirillum

which belongs to the class

Alphaproteobacteria

. The highest sequence similarity of strain JA318T was found with

Rhodospirillum sulfurexigens

JA143T (99.9 %). The DNA–DNA reassociation values of strain JA318T with Rsp. sulfurexigens JA143T and

Rhodospirillum photometricum

DSM 122T were 52±2 % and 45±1 %, respectively. The genomic DNA G+C content of strain JA318T was 60.2 mol%. Based on the morphological, physiological, chemotaxonomical and molecular evidence, strain JA318T is significantly different from the type strains of species of the genus

Rhodospirillum

, of the family

Rhodospirillaceae

, and it is proposed that the strain be classified as a representative of a novel species for which the name

Rhodospirillum

oryzae sp. nov. is proposed. The type strain is JA318T ( = KCTC 5960T = NBRC 107573T).
Winogradskyella

pulchriflava sp. nov., isolated from marine sediment
Authors: Kim So-Jeong , Choi Ye-Rin , Park Soo-Je , Kim Jong-Geol , Shin Kee-Sun , Roh Dong-Hyun , Rhee Sung-Keun
Published: February 2013

A taxonomic study was conducted on strain EM106T, isolated from a sediment sample of the East Sea, Republic of Korea. Comparative 16S rRNA gene sequence analyses showed that strain EM106T belongs to the family

Flavobacteriaceae

and is most closely related to

Winogradskyella echinorum

KMM 6211T and

Winogradskyella ulvae

KMM 6390T (97.8 and 97.3 % 16S rRNA gene sequence similarities, respectively). The G+C content of the genomic DNA of strain EM106T was 33.3 mol%, and the major respiratory quinone was menaquinone-6. The polar lipids of EM106T were phosphatidylethanolamine, two unidentified aminolipids and two unidentified lipids. DNA–DNA relatedness data indicated that strain EM106T represented a distinct species, separate from

W. echinorum

KMM 6211T and

W. ulvae

KMM 6390T. Strain EM106T possessed iso-C15 : 0, iso-C15 : 1 G and iso-C16 : 0 3-OH as the major cellular fatty acids. The isolate was Gram-staining-negative, strictly aerobic, short rod-shaped and motile by gliding. The strain grew at 10–35 °C (optimum, 25 °C), pH 6.5–9.0 (optimum, 7.5), and with 0.5–5 % (w/v) NaCl (optimum, 0.5–1 % NaCl). The overall physiological features of strain EM106T were very similar to those of

W. echinorum

KMM 6211T but only strain EM106T had nitrate reductase activity. On the basis of phenotypic and phylogenetic analyses, strain EM106T is proposed to represent a novel species,

Winogradskyella

pulchriflava. The type strain is EM106T( = KCTC 23858T = NCAIM B 02481T).
Enterococcus alcedinis sp. nov., isolated from common kingfisher (Alcedo atthis)
Authors: Frolková Petra , Švec Pavel , Sedláček Ivo , Mašlaňová Ivana , Černohlávková Jitka , Ghosh Anuradha , Zurek Ludek , Radiměřský Tomáš , Literák Ivan
Published: February 2013

Two Gram-positive, catalase-negative bacterial strains were isolated from the cloaca of common kingfishers (Alcedo atthis). Repetitive sequence-based PCR fingerprinting using the (GTG)5primer grouped these isolates into a single cluster separated from all known enterococcal species. The two strains revealed identical 16S rRNA gene sequences placing them within the genusEnterococcuswithEnterococcus aquimarinusLMG 16607Tas the closest relative (97.14 % similarity). Further taxonomic investigation using sequencing of the genes for the superoxide dismutase (sodA), phenylalanyl-tRNA synthase alpha subunit (pheS) and the RNA polymerase alpha subunit (rpoA) as well as application of whole-cell protein fingerprinting, automated ribotyping and extensive phenotyping confirmed that both strains belong to the same species. Based on data from this polyphasic study, these strains represent a novel species of the genusEnterococcus, for which the nameEnterococcusalcedinissp. nov. is proposed. The type strain is L34T( = CCM 8433T = LMG 27164T).
Cell-type specific inactivation of hippocampal CA1 disrupts location-dependent object recognition in the mouse
Authors: Haettig Jakob , Sun Yanjun , Wood Marcelo A. , Xu Xiangmin
Published: February 2013

The allatostatin receptor (AlstR)/ligand inactivation system enables potent regulation of neuronal circuit activity. To examine how different cell types participate in memory formation, we have used this system through Cre-directed, cell-type specific expression in mouse hippocampal CA1 in vivo and examined functional effects of inactivation of excitatory vs. inhibitory neurons on memory formation. We chose to use a hippocampus-dependent behavioral task involving location-dependent object recognition (LOR). The double transgenic mice, with the AlstRs selectively expressed in excitatory pyramidal neurons or inhibitory interneurons, were cannulated, targeting dorsal hippocampus to allow the infusion of the receptor ligand (the allatostatin [AL] peptide) in a time dependent manner. Compared to control animals, AL-infused animals showed no long-term memory for object location. While inactivation of excitatory or inhibitory neurons produced opposite effects on hippocampal circuit activity in vitro, the effects in vivo were similar. Both types of inactivation experiments resulted in mice exhibiting no long-term memory for object location. Together, these results demonstrate that the Cre-directed, AlstR-based system is a powerful tool for cell-type specific manipulations in a behaving animal and suggest that activity of either excitatory neurons or inhibitory interneurons is essential for proper long-term object location memory formation.
Effects of number of items and interval length on the acquisition of temporal order discrimination in radial maze in rats
Authors: Sugita Manami , Yamada Kazuo , Ichitani Yukio
Published: February 2013

Temporal order discrimination in rats was analyzed using a radial maze. The task consisted of a study phase in which two to five items (arms) were presented sequentially and, after a delay, a test phase in which two of these were simultaneously presented and the rat had to choose the arm presented earlier in the study phase. Acquisition of the task was better as the number of items in the study phase increased when the total study phase length was constant (4 min). Interval length between items in the study phase did not affect the acquisition regardless of the number of items. Additionally, discrimination performance was better when two items with more intervening items in the study phase (temporal lag) were used for the test. However, this tendency was obtained only when the last item of the study phase was included in the test pair. Results suggest that the number of items presented in the sequence, but not interval length between items, is an important factor in temporal order memory, and that a larger number of intervening items, as well as containing the last item in the study phase, contributes to the occurrence of the temporal lag effect.
Conflict between place and response navigation strategies: Effects on vicarious trial and error (VTE) behaviors
Authors: Schmidt Brandy , Papale Andrew , Redish A. David , Markus Etan J.
Published: February 2013

Navigation can be accomplished through multiple decision-making strategies, using different information-processing computations. A well-studied dichotomy in these decision-making strategies compares hippocampal-dependent “place” and dorsal-lateral striatal-dependent “response” strategies. A place strategy depends on the ability to flexibly respond to environmental cues, while a response strategy depends on the ability to quickly recognize and react to situations with well-learned action–outcome relationships. When rats reach decision points, they sometimes pause and orient toward the potential routes of travel, a process termed vicarious trial and error (VTE). VTE co-occurs with neurophysiological information processing, including sweeps of representation ahead of the animal in the hippocampus and transient representations of reward in the ventral striatum and orbitofrontal cortex. To examine the relationship between VTE and the place/response strategy dichotomy, we analyzed data in which rats were cued to switch between place and response strategies on a plus maze. The configuration of the maze allowed for place and response strategies to work competitively or cooperatively. Animals showed increased VTE on trials entailing competition between navigational systems, linking VTE with deliberative decision-making. Even in a well-learned task, VTE was preferentially exhibited when a spatial selection was required, further linking VTE behavior with decision-making associated with hippocampal processing.
Nicotine modulates the long-lasting storage of fear memory
Authors: Lima Ramón H. , Radiske Andressa , Köhler Cristiano A. , Gonzalez Maria Carolina , Bevilaqua Lia R. , Rossato Janine I. , Medina Jorge H. , Cammarota Martín
Published: February 2013

Late post-training activation of the ventral tegmental area (VTA)–hippocampus dopaminergic loop controls the entry of information into long-term memory (LTM). Nicotinic acetylcholine receptors (nAChR) modulate VTA function, but their involvement in LTM storage is unknown. Using pharmacological and behavioral tools, we found that α7-nAChR-mediated cholinergic interactions between the pedunculopontine tegmental nucleus and the medial prefrontal cortex modulate the duration of fear-motivated memories, maybe by regulating the activation state of VTA–hippocampus dopamine connections.
The Pseudomonas syringae Effector HopQ1 Promotes Bacterial Virulence and Interacts with Tomato 14-3-3 Proteins in a Phosphorylation-Dependent Manner
Authors: Li Wei , Yadeta Koste A. , Elmore James Mitch , Coaker Gitta
Published: February 2013

A key virulence strategy of bacterial pathogens is the delivery of multiple pathogen effector proteins into host cells during infection. The Hrp outer protein Q (HopQ1) effector from Pseudomonas syringae pv tomato (Pto) strain DC3000 is conserved across multiple bacterial plant pathogens. Here, we investigated the virulence function and host targets of HopQ1 in tomato (Solanum lycopersicum). Transgenic tomato lines expressing dexamethasone-inducible HopQ1 exhibited enhanced disease susceptibility to virulent Pto DC3000, the Pto ƊhrcC mutant, and decreased expression of a pathogen-associated molecular pattern-triggered marker gene after bacterial inoculation. HopQ1-interacting proteins were coimmunoprecipitated and identified by mass spectrometry. HopQ1 can associate with multiple tomato 14-3-3 proteins, including TFT1 and TFT5. HopQ1 is phosphorylated in tomato, and four phosphorylated peptides were identified by mass spectrometry. HopQ1 possesses a conserved mode I 14-3-3 binding motif whose serine-51 residue is phosphorylated in tomato and regulates its association with TFT1 and TFT5. Confocal microscopy and fractionation reveal that HopQ1 exhibits nucleocytoplasmic localization, while HopQ1 dephosphorylation mimics exhibit more pronounced nuclear localization. HopQ1 delivered from Pto DC3000 was found to promote bacterial virulence in the tomato genotype Rio Grande 76R. However, the HopQ1(S51A) mutant delivered from Pto DC3000 was unable to promote pathogen virulence. Taken together, our data demonstrate that HopQ1 enhances bacterial virulence and associates with tomato 14-3-3 proteins in a phosphorylation-dependent manner that influences HopQ1’s subcellular localization and virulence-promoting activities in planta.
Production of a High-Efficiency TILLING Population through Polyploidization
Authors: Tsai Helen , Missirian Victor , Ngo Kathie J. , Tran Robert K. , Chan Simon R. , Sundaresan Venkatesan , Comai Luca
Published: February 2013

Targeting Induced Local Lesions in Genomes (TILLING) provides a nontransgenic method for reverse genetics that is widely applicable, even in species where other functional resources are missing or expensive to build. The efficiency of TILLING, however, is greatly facilitated by high mutation density. Species vary in the number of mutations induced by comparable mutagenic treatments, suggesting that genetic background may affect the response. Allopolyploid species have often yielded higher mutation density than diploids. To examine the effect of ploidy, we autotetraploidized the Arabidopsis (Arabidopsis thaliana) ecotype Columbia, whose diploid has been used for TILLING extensively, and mutagenized it with 50 mm ethylmethane sulfonate. While the same treatment sterilized diploid Columbia, the tetraploid M1 plants produced good seed. To determine the mutation density, we searched 528 individuals for induced mutations in 15 genes for which few or no knockout alleles were previously available. We constructed tridimensional pools from the genomic DNA of M2 plants, amplified target DNA, and subjected them to Illumina sequencing. The results were analyzed with an improved version of the mutation detection software CAMBa that accepts any pooling scheme. This small population provided a rich resource with approximately 25 mutations per queried 1.5-kb fragment, including on average four severe missense and 1.3 truncation mutations. The overall mutation density of 19.4 mutations Mb–1 is 4 times that achieved in the corresponding diploid accession, indicating that genomic redundancy engenders tolerance to high mutation density. Polyploidization of diploids will allow the production of small populations, such as less than 2,000, that provide allelic series from knockout to mild loss of function for virtually all genes.
The Regulatory Properties of Rubisco Activase Differ among Species and Affect Photosynthetic Induction during Light Transitions
Authors: Carmo-Silva A. Elizabete , Salvucci Michael E.
Published: February 2013

Rubisco’s catalytic chaperone, Rubisco activase (Rca), uses the energy from ATP hydrolysis to restore catalytic competence to Rubisco. In Arabidopsis (Arabidopsis thaliana), inhibition of Rca activity by ADP is fine tuned by redox regulation of the α-isoform. To elucidate the mechanism for Rca regulation in species containing only the redox-insensitive β-isoform, the response of activity to ADP was characterized for different Rca forms. When assayed in leaf extracts, Rubisco activation was significantly inhibited by physiological ratios of ADP to ATP in species containing both α-Rca and β-Rca (Arabidopsis and camelina [Camelina sativa]) or just the β-Rca (tobacco [Nicotiana tabacum]). However, Rca activity was insensitive to ADP inhibition in an Arabidopsis transformant, rwt43, which expresses only Arabidopsis β-Rca, although not in a transformant of Arabidopsis that expresses a tobacco-like β-Rca. ATP hydrolysis by recombinant Arabidopsis β-Rca was much less sensitive to inhibition by ADP than recombinant tobacco β-Rca. Mutation of 17 amino acids in the tobacco β-Rca to the corresponding Arabidopsis residues reduced ADP sensitivity. In planta, Rubisco deactivated at low irradiance except in the Arabidopsis rwt43 transformant containing an ADP-insensitive Rca. Induction of CO2 assimilation after transition from low to high irradiance was much more rapid in the rwt43 transformant compared with plants containing ADP-sensitive Rca forms. The faster rate of photosynthetic induction and a greater enhancement of growth under a fluctuating light regime by the rwt43 transformant compared with wild-type Arabidopsis suggests that manipulation of Rca regulation might provide a strategy for enhancing photosynthetic performance in certain variable light environments.
Altered Lipid Composition and Enhanced Nutritional Value of Arabidopsis Leaves following Introduction of an Algal Diacylglycerol Acyltransferase 2
Authors: Sanjaya  , Miller Rachel , Durrett Timothy P. , Kosma Dylan K. , Lydic Todd A. , Muthan Bagyalakshmi , Koo Abraham J.K. , Bukhman Yury V. , Reid Gavin E. , Howe Gregg A. , Ohlrogge John , Benning Christoph
Published: February 2013

Enhancement of acyl-CoA–dependent triacylglycerol (TAG) synthesis in vegetative tissues is widely discussed as a potential avenue to increase the energy density of crops. Here, we report the identification and characterization of Chlamydomonas reinhardtii diacylglycerol acyltransferase type two (DGTT) enzymes and use DGTT2 to alter acyl carbon partitioning in plant vegetative tissues. This enzyme can accept a broad range of acyl-CoA substrates, allowing us to interrogate different acyl pools in transgenic plants. Expression of DGTT2 in Arabidopsis thaliana increased leaf TAG content, with some molecular species containing very-long-chain fatty acids. The acyl compositions of sphingolipids and surface waxes were altered, and cutin was decreased. The increased carbon partitioning into TAGs in the leaves of DGTT2-expressing lines had little effect on transcripts of the sphingolipid/wax/cutin pathway, suggesting that the supply of acyl groups for the assembly of these lipids is not transcriptionally adjusted. Caterpillars of the generalist herbivore Spodoptera exigua reared on transgenic plants gained more weight. Thus, the nutritional value and/or energy density of the transgenic lines was increased by ectopic expression of DGTT2 and acyl groups were diverted from different pools into TAGs, demonstrating the interconnectivity of acyl metabolism in leaves.
Coordination of the Vernalization Response through a VIN3 and FLC Gene Family Regulatory Network in Arabidopsis
Authors: Kim Dong-Hwan , Sung Sibum
Published: February 2013

Vernalization is an environmentally induced epigenetic switch in which winter cold triggers epigenetic silencing of floral repressors and thus provides competence to flower in spring. Vernalization triggers the recruitment of chromatin-modifying complexes to a clade of flowering repressors that are epigenetically silenced via chromatin modifications. In Arabidopsis thaliana, VERNALIZATION INSENSITIVE3 (VIN3) and its related plant homeodomain finger proteins act together with Polycomb Repressive Complex 2 to increase repressive histone marks at floral repressor loci, including FLOWERING LOCUS C (FLC) and its related genes, by vernalization. Here, we show that VIN3 family of proteins nonredundantly functions to repress different subsets of the FLC gene family during the course of vernalization. Each VIN3 family protein binds to modified histone peptides in vitro and directly associates with specific sets of FLC gene family chromatins in vivo to mediate epigenetic silencing. In addition, members of the FLC gene family are also differentially regulated during the course of vernalization to mediate proper vernalization response. Our results show that these two gene families cooperated during the course of evolution to ensure proper vernalization response through epigenetic changes.
Educational Reforms Set Professional Boundaries: The Spanish Audit Function, 1850–1988
Authors: Carrera Nieves , Carmona Salvador
Published: February 2013

Research on the accounting profession considers the notion of boundaries a useful, albeit wide and elusive, concept. However, recent sociological and organizational research has sharpened the concept of boundaries to enable further examination of relational processes. In this paper, we draw on this research to advance our understanding of the process of constructing boundaries in auditing. In doing so, we focus on the unique institutional conditions in Spain during the period 1850–1988. This period witnessed the overwhelming role of the State in any sphere of economic and social life. In the case of auditing, the influence of the State was augmented as the academic background became the site of influence, which resulted in educational reforms setting professional boundaries. Our findings provide some additional insights into the boundaries construction process and the dynamics of the symbolic and social boundaries contingent on the wider contexts of the audit function.
An online Symptom Care and Management System to monitor and support patients receiving chemotherapy: A pilot study
Authors: Chan Moon‐Fai , Ang Emily , Duong Minh Cam , Chow Ying Leng
Published: February 2013

This pilot study aimed to explore the benefits of an online Symptom Care and Management System (SCMS) in the home monitoring and symptom management of patients receiving chemotherapy. A single‐group experimental design was employed, and four patients aged 39–59 years undergoing their first chemotherapy treatment were recruited from one cancer centre in Singapore from November 2010 to February 2011. A mixed method was used to collect patient's data. All patients used the SCMS to record and send daily symptom reports to the cancer centre and receive symptom management advice from the nurse via teleconferencing. Patients' perceptions on the use of the system were evaluated. All participants believed that the system improved the management of their symptom and felt reassured they were being monitored at home. The results presented in this pilot study suggest that the SCMS has the potential to enhance the remote monitoring, feasible and acceptable way for a specific group of cancer patients to manage their symptom at home.
The learning curve for laparoscopic extended pelvic lymphadenectomy for intermediate‐ and high‐risk prostate cancer: implications for compliance with existing guidelines
Authors: Eden Christopher G. , Zacharakis Evangelos , Bott Simon
Published: February 2013

Objective

To investigate the learning curve for performing extended pelvic lymphadenectomy (ePLND) during laparoscopic radical prostatectomy (LRP) in patients with intermediate‐ and high‐risk prostate cancer.

Patients and methods

In all, 500 patients underwent ePLND for intermediate‐ or high‐risk prostate cancer by one surgeon during a 48‐month period. A transperitoneal laparoscopic approach was used in all patients to allow adequate access to the internal iliac vessels.

The variables chosen as being the most important discriminators of the quality of ePLND were operating time, complication rate and lymph node (LN) yield.

The learning curves for ePLND were calculated using the cumulative sum and cumulative average methods and the number of procedures performed until attainment of acceptable failure rates (competence levels) was calculated. LN parameters were compared with the results from the preceding 311 cases where limited PLND was undertaken.

Results

The median (range) preoperative PSA level was 8.0(1–62.5) ng/mL and biopsy Gleason score was 7(6–10). In all, 64% of patients had intermediate‐risk and 36% had high‐risk prostate cancer. There were no intraoperative blood transfusions and no conversions to open surgery. The median (range) blood loss was 200(10–1400) mL and the postoperative transfusion rate was 1.6%.

The operating time fell at a steady rate of 2.7% after the 15th case and plateaued after 130 patients. At competence levels of 5% and 10%, the learning curve for all complications ended after 346 and 136 patients, respectively.

At a 5% competence level the learning curve for PLND‐specific complications was 40 cases and there was no learning curve at a 10% competence level.

The overall complication rate was 7.2% of which almost half (47%) were deemed to be PLND‐specific. The cumulative average of the LN counts plateaued after 150 procedures. Furthermore, the median LN count after ePLND was more than double that of the authors' historical standard PLND controls (14 vs 6, P < 0.001) and increased with experience up to the end of the series (9 to 20).

The likelihood of LN involvement (LNI) correlated with biopsy and pathological Gleason grade, clinical and pathological stage and d'Amico risk group.

Conclusions

This study suggests a learning curve of ≈130 cases for operating time, 136 cases for all complications, 40 cases for PLND‐specific complications and 150 cases for LN yield.

The risk of LNI for patients with intermediate‐ and high‐risk prostate cancer was 8.4% and 19.4%, respectively, which suggests that a significant proportion would benefit from ePLND. It also shows that ePLND can be safely incorporated into LRP, and therefore also into robot‐assisted RP, in a high‐volume setting.
Catalase gene C‐262T polymorphism: Importance in ulcerative colitis
Authors: Khodayari Siamak , Salehi Zivar , Fakhrieh Asl Saba , Aminian Keyvan , Mirzaei Gisomi Nadia , Torabi Dalivandan Saeideh
Published: February 2013

Background and Aim

Reactive oxygen species (ROS) are by‐products of the cellular metabolism and have important roles in the normal physiology of the cell. However, when ROS production exceeds the antioxidant capacity, a state known as oxidative stress, damage to cellular macromolecules emerges. A crucial role in counteracting ROS is played by the enzyme catalase. A common polymorphism in the catalase (CAT) promoter region (C‐262T) alters the expression as well as blood catalase levels, and leads to a number of human diseases. Ulcerative colitis (UC) is an inflammatory condition of the large bowel that is known to be influenced by oxidative stress. In this study, we aimed to evaluate the association of CAT C‐262T polymorphism on the risk of UC.

Methods

Samples were collected from 60 patients diagnosed with UC and 78 control subjects, and genotyped by allele‐specific polymerase chain reaction.

Results

We found that CAT C‐262T genotype frequencies were significantly different between cases and controls (P = 0.002). Individuals carrying the −262C/T genotype had a greater risk for UC compared with C/C genotype (odds ratio, 4.88; 95% confidence interval, 1.73–13.75, P = 0.002).

Conclusions

This study indicates that CAT C‐262T polymorphism may be associated with UC, and that the −262C/T genotype may be a risk factor for the disease. Further studies are needed to confirm the results.
Charismatic Rhetoric in the 2008 Presidential Campaign: Commonalities and Differences
Authors: Schroedel Jean , Bligh Michelle , Merolla Jennifer , Gonzalez Randall
Published: February 2013

For all of the attention given to charisma in media accounts of presidential campaigns, there is surprisingly little systematic research on the use and effect of charismatic rhetoric, even though researchers outside of political science have attempted to deconstruct and analyze the different components of rhetorical speech. We draw from this scholarship to explore the use of different forms of charismatic rhetoric throughout the entire 2008 presidential campaign season. We use DICTION 5.0, a content analysis program designed specifically for political discourse, to explore the use of rhetoric in the campaign. We begin by mapping the prevalence of different types of rhetoric and then move on to examine the impact of partisanship and electoral context (primary vs. general election) on rhetorical choices.
Stimulation of Naïve Monocytes and PBMCs with Coagulation Proteases Results in Thrombin‐Mediated and PAR‐1‐Dependent Cytokine Release and Cell Proliferation in PBMCs Only
Authors: Nieuwenhuizen L. , Falkenburg W. J. J. , Schutgens R. E. G. , Roosendaal G. , van Veghel K. , Biesma D. H. , Lafeber F. P. J. G.
Published: February 2013

Protease‐activated receptors (PARs) are stimulated by proteolytic cleavage of their extracellular domain. Coagulation proteases, such as FVIIa, the binary TF‐FVIIa complex, free FXa, the ternary TF‐FVIIa‐FXa complex and thrombin, are able to stimulate PARs. Whereas the role of PARs on platelets is well known, their function in naïve monocytes and peripheral blood mononuclear cells (PBMCs) is largely unknown. This is of interest because PAR‐mediated interactions of coagulation proteases with monocytes and PBMCs in diseases with an increased activation of coagulation may promote inflammation. To evaluate PAR‐mediated inflammatory reactions in naïve monocytes and PBMCs stimulated with coagulation proteases. For this, PAR expression at protein and RNA level on naïve monocytes and PBMCs was evaluated with flow cytometry and RT‐PCR. In addition, cytokine release (IL‐1β, IL‐6, IL‐8, IL‐10, TNF‐α) in stimulated naïve and PBMC cell cultures was determined. In this study, it is demonstrated that naïve monocytes express all four PARs at the mRNA level, and PAR‐1, ‐3 and ‐4 at the protein level. Stimulation of naïve monocytes with coagulation proteases did not result in alterations in PAR expression or in the induction of inflammation involved cytokines like interleukin‐1β (IL‐1β), interleukin‐6 (IL‐6), interleukin‐8, interleukin‐10 or tumour necrosis factor‐α. In contrast, stimulation of PBMCs with coagulation proteases resulted in thrombin‐mediated induction of IL‐1β and IL‐6 cytokine production and PBMC cell proliferation in a PAR‐1‐dependent manner. These data demonstrate that naïve monocytes are not triggered by coagulation proteases, whereas thrombin is able to elicit pro‐inflammatory events in a PAR‐1‐dependent manner in PBMCs.
Tonic muscle pain does not increase fusimotor drive to human leg muscles: implications for chronic muscle pain
Authors: Fazalbhoy Azharuddin , Macefield Vaughan G. , Birznieks Ingvars
Published: February 2013

New Findings

What is the central question of this study?

Based on data obtained from experimental animals, muscle pain is believed to cause a reflex activation of fusimotor neurones and thereby increase the sensitivity of muscle spindles to stretch. Using a model of long‐lasting muscle pain, we asked the question: does tonic muscle pain increase the resting discharge of muscle spindles in human subjects?

What is the main finding and its importance?

Microelectrode recordings from single muscle spindle afferents revealed no net change in the discharge of spontaneously active spindle endings during moderate–strong pain lasting ∼1 h. We conclude that, unlike the situation in anaesthetized animals, muscle pain does not cause a reflex increase in fusimotor drive and spindle discharge.

Experimental pain induced in animals has shown that noxious stimulation of group III and IV afferents increases the firing of muscle spindles via a reflex excitation of fusimotor (γ) motoneurones. Chronic muscle pain has been hypothesized to develop as a result of a vicious cycle involving this mechanism. In order to explore the effects of long‐lasting muscle pain on the fusimotor system, single unit muscle spindle afferents were recorded from 15 subjects. Afferent activity was recorded from foot and ankle extensor muscles whilst infusing hypertonic saline into the tibialis anterior muscle of the ipsilateral leg, producing moderate–strong pain lasting for ∼60 min. A change in fusimotor drive was inferred by observing changes in the mean discharge rate of spontaneously active muscle spindle afferents. Homonymous and heteronymous muscles remained relaxed and showed no increase in activity, arguing against any fusimotor‐driven increase in motor activity, and there was no net change in the firing of muscle spindle afferents. We conclude that long‐lasting stimulation of group III and IV afferents fails to excite fusimotor neurones and increase muscle spindle discharge. Accordingly, the vicious cycle theory has no functional basis for the development of myalgia in human subjects.
Metabolic Responses of Saccharomyces cerevisiae to Valine and Ammonium Pulses during Four-Stage Continuous Wine Fermentations
Authors: Clement T. , Perez M. , Mouret J. R. , Sanchez I. , Sablayrolles J. M. , Camarasa C.
Published: February 2013

Nitrogen supplementation, which is widely used in winemaking to improve fermentation kinetics, also affects the products of fermentation, including volatile compounds. However, the mechanisms underlying the metabolic response of yeast to nitrogen additions remain unclear. We studied the consequences forSaccharomyces cerevisiaemetabolism of valine and ammonium pulses during the stationary phase of four-stage continuous fermentation (FSCF). This culture technique provides cells at steady state similar to that of the stationary phase of batch wine fermentation. Thus, the FSCF device is an appropriate and reliable tool for individual analysis of the metabolic rerouting associated with nutrient additions, in isolation from the continuous evolution of the environment in batch processes. Nitrogen additions, irrespective of the nitrogen-containing compound added, substantially modified the formation of fermentation metabolites, including glycerol, succinate, isoamyl alcohol, propanol, and ethyl esters. This flux redistribution, fulfilling the requirements for precursors of amino acids, was consistent with increased protein synthesis resulting from increased nitrogen availability. Valine pulses, less efficient than ammonium addition in increasing the fermentation rate, were followed by a massive conversion of this amino acid in isobutanol and isobutyl acetate through the Ehrlich pathway. However, additional routes were involved in valine assimilation when added in stationary phase. Overall, we found that particular metabolic changes may be triggered according to the nature of the amino acid supplied, in addition to the common response. Both these shared and specific modifications should be considered when designing strategies to modulate the production of volatile compounds, a current challenge for winemakers.
Light-Dependent Functions of the Fusarium fujikuroi CryD DASH Cryptochrome in Development and Secondary Metabolism
Authors: Castrillo Marta , García-Martínez Jorge , Avalos Javier
Published: February 2013

DASH (Drosophila,Arabidopsis,Synechocystis, human) cryptochromes (cry-DASHs) constitute a subgroup of the photolyase cryptochrome family with diverse light-sensing roles, found in most taxonomical groups. The genome ofFusarium fujikuroi, a phytopathogenic fungus with a rich secondary metabolism, contains a gene encoding a putative cry-DASH, named CryD. The expression of thecryDgene is induced by light in the wild type, but not in mutants of the “white collar” genewcoA. Targeted ΔcryDmutants show light-dependent phenotypic alterations, including changes in morphology and pigmentation, which disappear upon reintroduction of a wild-typecryDallele. In addition to microconidia, the colonies of the ΔcryDmutants produced under illumination and nitrogen starvation large septated spores called macroconidia, absent in wild-type colonies. The ΔcryDmutants accumulated similar amounts of carotenoids to the control strain under constant illumination, but produced much larger amounts of bikaverin under nitrogen starvation, indicating a repressing role for CryD in this biosynthetic pathway. Additionally, a moderate photoinduction of gibberellin production was exhibited by the wild type but not by the ΔcryDmutants. The phenotypic alterations of the ΔcryDmutants were only noticeable in the light, as expected from the low expression ofcryDin the dark, but did not correlate with mRNA levels for structural genes of the bikaverin or gibberellin biosynthetic pathways, suggesting the participation of CryD in posttranscriptional regulatory mechanisms. This is the first report on the participation of a cry-DASH protein in the regulation of fungal secondary metabolism.
Evaluation of Bovine Feces-Associated Microbial Source Tracking Markers and Their Correlations with Fecal Indicators and Zoonotic Pathogens in a Brisbane, Australia, Reservoir
Authors: Ahmed W. , Sritharan T. , Palmer A. , Sidhu J. P. S. , Toze S.
Published: February 2013

This study was aimed at evaluating the host specificity and host sensitivity of two bovine feces-associated bacterial (BacCow-UCD and cowM3) and one viral [bovine adenovirus (B-AVs)] microbial source tracking (MST) markers by screening 130 fecal and wastewater samples from 10 target and nontarget host groups in southeast Queensland, Australia. In addition, 36 water samples were collected from a reservoir and tested for the occurrence of all three bovine feces-associated markers along with fecal indicator bacteria (FIB),
Campylobacter
spp.,
Escherichia coli
O157, and
Salmonella
spp. The overall host specificity values of the BacCow-UCD, cowM3, and B-AVs markers to differentiate between bovine and other nontarget host groups were 0.66, 0.88, and 1.00, respectively (maximum value of 1.00). The overall host sensitivity values of these markers, however, in composite bovine wastewater and individual bovine fecal DNA samples were 0.93, 0.90, and 0.60, respectively (maximum value of 1.00). Among the 36 water samples tested, 56%, 22%, and 6% samples were PCR positive for the BacCow-UCD, cowM3, and B-AVs markers, respectively. Among the 36 samples tested, 50% and 14% samples were PCR positive for the
Campylobacter
16S rRNA and
E. coli
O157
rfbE
genes, respectively. Based on the results, we recommend that multiple bovine feces-associated markers be used if possible for bovine fecal pollution tracking. Nonetheless, the presence of the multiple bovine feces-associated markers along with the presence of potential zoonotic pathogens indicates bovine fecal pollution in the reservoir water samples. Further research is required to understand the decay rates of these markers in relation to FIB and zoonotic pathogens.
Utilization of Virus ϕCh1 Elements To Establish a Shuttle Vector System for Halo(alkali)philic Archaea via Transformation of Natrialba magadii
Authors: Mayrhofer-Iro M. , Ladurner A. , Meissner C. , Derntl C. , Reiter M. , Haider F. , Dimmel K. , Rössler N. , Klein R. , Baranyi U. , Scholz H. , Witte A.
Published: February 2013

In the study described here, we successfully developed a transformation system for halo(alkali)philic members of the
Archaea
. This transformation system comprises a series of
Natrialba magadii
/
Escherichia coli
shuttle vectors based on a modified method to transform halophilic members of the
Archaea
and genomic elements of the
N. magadii
virus ϕCh1. The shuttle vector pRo-5, based on the
repH
-containing region of ϕCh1, stably replicated in
E. coli
and
N. magadii
and in several halophilic and haloalkaliphilic members of the
Archaea
not transformable so far. The ϕCh1 operon ORF53/ORF54 (
repH
) was essential for pRo-5 replication and was thus identified as the minimal replication origin. The plasmid allowed homologous and heterologous gene expression, as exemplified by the expression of ϕCh1 ORF34
52
, which encodes a structural protein, and the reporter gene
bgaH
of
Haloferax lucentense
in
N. magadii
. The new transformation/vector system will facilitate genetic studies within
N. magadii
and other haloalkaliphilic archaea and will allow the detailed characterization of the gene functions of
N. magadii
virus ϕCh1 in their extreme environments.
Revisiting Methanotrophic Communities in Sewage Treatment Plants
Authors: Ho Adrian , Vlaeminck Siegfried E. , Ettwig Katharina F. , Schneider Bellinda , Frenzel Peter , Boon Nico
Published: February 2013

The methanotrophic potential in sewage treatment sludge was investigated. We detected a diverse aerobic methanotrophic community that potentially plays a significant role in mitigating methane emission in this environment. The results suggest that community structure was determined by conditions specific to the processes in a sewage treatment plant.