bluuebunny/crossref_metadata_2025_split huggingface dataset:
Development of a bipartite ecdysone‐responsive gene switch for the oomycete Phytophthora infestans and its use to manipulate transcription during axenic culture and plant infection
Authors: Gamboa‐Meléndez Heber , Judelson Howard S. 
Published: May 2014

SummaryConditional expression systems have been proven to be useful tools for the elucidation of gene function in many taxa. Here, we report the development of the first useful inducible promoter system for an oomycete, based on an ecdysone receptor (EcR) and the ecdysone analogue methoxyfenozide. In Phytophthora infestans, the potato late blight pathogen, a monopartite transactivator containing the VP16 activation domain from herpes simplex virus, the GAL4 DNA‐binding domain from yeast and the EcR receptor domain from the spruce budworm enabled high levels of expression of a β‐glucuronidase (GUS) reporter gene, but unacceptable basal activity in the absence of the methoxyfenozide inducer. Greatly improved performance was obtained using a bipartite system in which transcription is activated by a heterodimer between a chimera of VP16 and the migratory locust retinoid X receptor, and a separate EcR‐DNA‐binding domain chimera. Transformants were obtained that exhibited >100‐fold activation of the reporter by methoxyfenozide, with low basal levels of expression and induced activity approaching that of the strong ham34 promoter. Performance varied between transformants, probably as a result of position effects. The addition of methoxyfenozide enabled strong induction during hyphal growth, zoosporogenesis and colonization of tomato. No significant effects of the inducer or transactivators on growth, development or pathogenicity were observed. The technology should therefore be a useful addition to the arsenal of methods for the study of oomycete plant pathogens.
Regional data analysis of Candida non‐albicans strains collected in United States medical sites over a 6‐year period, 2006–2011
Authors: Pfaller Michael A. , Jones Ronald N. , Castanheira Mariana 
Published: May 2014

SummaryLimited data are available on temporal and geographic variation of occurrence and antifungal resistance of non‐C. albicans Candida species (non‐CA‐CSP) from the USA. The objective of this study was to evaluate the occurrence and antifungal resistance of 1694 isolates of non‐CA‐CSP collected during the period 2006–2011. Isolates were recovered in 33 hospitals located in four regions: Northcentral, North‐east, South‐east and West and tested using CLSI reference broth microdilution methods. Non‐CA‐CSP represented 55.6% of all Candida. C. glabrata was most predominant (39–42% of non‐CA‐CSP). Infections due to C. glabrata, C. krusei and C. dubliniensis increased over the 6 years. Anidulafungin (3.6%) and caspofungin (5.7%) resistance were prominent among C. glabrata from the North‐east and West regions respectively. Resistance to micafungin was detected in 2.0% and 2.9% of C. glabrata from the West and North‐east regions respectively. Echinocandin resistance was low, except for C. dubliniensis. Azole resistance was most prominent among C. glabrata from the South‐east (13.6% fluconazole R) and the West (18.0%). Cross‐resistance among three tested azoles was observed in C. glabrata from all regions. Whereas differences in species distribution and antifungal R varied across geographic regions, there was little evidence of temporal increase in resistance to azoles or echinocandins in the monitored non‐CA‐CSP.
Altered structural covariance of the striatum in functional dyspepsia patients
Authors: Liu P. , Zeng F. , Yang F. , Wang J. , Liu X. , Wang Q. , Zhou G. , Zhang D. , Zhu M. , Zhao R. , Wang A. , Gong Q. , Liang F. 
Published: May 2014

AbstractBackgroundFunctional dyspepsia (FD) is thought to be involved in dysregulation within the brain–gut axis. Recently, altered striatum activation has been reported in patients with FD. However, the gray matter (GM) volumes in the striatum and structural covariance patterns of this area are rarely explored. The purpose of this study was to examine the GM volumes and structural covariance patterns of the striatum between FD patients and healthy controls (HCs).MethodsT1‐weighted magnetic resonance images were obtained from 44 FD patients and 39 HCs. Voxel‐based morphometry (VBM) analysis was adopted to examine the GM volumes in the two groups. The caudate‐ or putamen‐related regions identified from VBM analysis were then used as seeds to map the whole brain voxel‐wise structural covariance patterns. Finally, a correlation analysis was used to investigate the effects of FD symptoms on the striatum.Key ResultsThe results showed increased GM volumes in the bilateral putamen and right caudate. Compared with the structural covariance patterns of the HCs, the FD‐related differences were mainly located in the amygdala, hippocampus/parahippocampus (HIPP/paraHIPP), thalamus, lingual gyrus, and cerebellum. And significant positive correlations were found between the volumes in the striatum and the FD duration in the patients.Conclusions & InferencesThese findings provided preliminary evidence for GM changes in the striatum and different structural covariance patterns in patients with FD. The current results might expand our understanding of the pathophysiology of FD.
The link between negative affect, vagal tone, and visceral sensitivity in quiescent Crohn's disease
Authors: Rubio A. , Pellissier S. , Picot A. , Dantzer C. , Bonaz B. 
Published: May 2014

AbstractAutonomic dysfunction and mood disorders are frequently described in Crohn's disease (CD) and are known to influence visceral sensitivity. We addressed the link between vagal tone, negative affect, and visceral sensitivity in CD patients without concomitant features of irritable bowel syndrome (IBS). Rectal distensions to a discomfort threshold of 70% and onset of pain were performed in nine CD patients in remission and eight healthy controls. Autonomic parameters were evaluated with heart rate variability and electrodermal reactivity. We showed that CD patients had (i) higher scores of depressive symptomatology (12 ± 3 in patients vs 4 ± 1 in controls on the Center for Epidemiologic Studies‐Depression Scale; p = 0.038), (ii) reduced vagal tone (HF 257 ± 84 ms2 vs 1607 ± 1032 ms2, p = 0.043; LF 455 ± 153 ms2 vs 1629 ± 585 ms2, p = 0.047), (iii) decreased sympathetic reactivity during an aversive stimulus, and (iv) higher tolerance to rectal distension pressures (43 ± 3 mmHg vs 30 ± 2 mmHg, p = 0.002) and low sensitivity index scores. In conclusion, our results provide preliminary evidence that patients with quiescent CD, in the absence of IBS, are hyposensate to experimental rectal distension. These data provide further evidence that anxiety and depressive symptomatology in addition to autonomic dysfunction modulate visceral pain perception in quiescent CD patients in the absence of IBS.
Giardia lamblia binding immunoglobulin protein triggers maturation of dendritic cells via activation of TLR4‐MyD88‐p38 and ERK1/2 MAPKs
Authors: Lee H.‐Y. , Kim J. , Noh H. J. , Kim H.‐P. , Park S.‐J. 
Published: May 2014

SummaryMuch remains unknown about the mammalian immune response to Giardia lamblia, a protozoan pathogen that causes diarrhoeal outbreaks. We fractionated protein extracts of G. lamblia trophozoites by Viva‐spin centrifugation, DEAE ion exchange and gel filtration chromatography. Resultant fractions were screened for antigenic molecules by western blots analysis using anti‐G. lamblia antibodies (Abs), resulting in identification of G. lamblia binding immunoglobulin protein (GlBiP). Maturation of mouse dendritic cells (DCs) in response to recombinant GlBiP (rGlBiP) was detected by increased expression of surface molecules such as CD80, CD86 and MHC class II; these mature DCs, produced pro‐inflammatory cytokines (TNF‐α, IL‐12 and IL‐6). Especially, the truncated rGlBiP containing the heat‐shock protein 70 domain‐induced cytokine production from mouse DCs. rGlBiP‐induced DC activation was initiated by TLR4 in a MyD88‐dependent way and occurred through activation of p38 and ERK1/2 MAPKs as well as increased activity of NF‐κB and AP‐1. Moreover, CD4+ T cells stimulated with rGlBiP‐treated DCs produced high levels of IL‐2 and IFN‐γ. Together, our results suggest that GlBiP contributes to maturation of DCs via activation of TLR4‐MyD88‐p38, ERK1/2 MAPK, NF‐κB and AP‐1.
How do nurses in palliative care perceive the concept of self‐image?
Authors: Jeppsson Margareth , Thomé Bibbi 
Published: May 2014

BackgroundNursing research indicates that serious illness and impending death influence the individual's self‐image. Few studies define what self‐image means. Thus it seems to be urgent to explore how nurses in palliative care perceive the concept of self‐image, to get a deeper insight into the concept′s applicability in palliative care.AimTo explore how nurses in palliative care perceive the concept of self‐image.DesignQualitative descriptive design.MethodIn‐depth interviews with 17 nurses in palliative care were analysed using phenomenography. The study gained ethical approval.ResultsThe concept of self‐image was perceived as both a familiar and an unfamiliar concept. Four categories of description with a gradually increasing complexity were distinguished: Identity, Self‐assessment, Social function and Self‐knowledge. They represent the collective understanding of the concept and are illustrated in a ‘self‐image map’. The identity‐category emerged as the most comprehensive one and includes the understanding of ‘Who I am’ in a multidimensional way.ConclusionThe collective understanding of the concept of self‐image include multi‐dimensional aspects which not always were evident for the individual nurse. Thus, the concept of self‐image needs to be more verbalised and reflected on if nurses are to be comfortable with it and adopt it in their caring context. The ‘self‐image map’ can be used in this reflection to expand the understanding of the concept. If the multi‐dimensional aspects of the concept self‐image could be explored there are improved possibilities to make identity‐promoting strategies visible and support person‐centred care.
The GUN4 protein plays a regulatory role in tetrapyrrole biosynthesis and chloroplast‐to‐nucleus signalling in Chlamydomonas reinhardtii
Authors: Brzezowski Pawel , Schlicke Hagen , Richter Andreas , Dent Rachel M. , Niyogi Krishna K. , Grimm Bernhard 
Published: May 2014

SummaryThe GENOMES UNCOUPLED 4 (GUN4) protein is found only in aerobic photosynthetic organisms. We investigated the role of GUN4 in metabolic activities of the Mg branch of the tetrapyrrole biosynthesis pathway and the plastid signal‐mediated changes of nuclear gene expression in Chlamydomonas reinhardtii. In light, gun4 accumulates only 40% of the wild‐type chlorophyll level. Light‐ or dark‐grown gun4 mutant accumulates high levels of protoporphyrin IX (Proto), and displays increased sensitivity to moderate light intensities. Despite the photooxidative stress, gun4 fails to downregulate mRNA levels of the tetrapyrrole biosynthesis and the photosynthesis‐associated nuclear genes (PhANGs). In contrast, upon illumination, the Proto‐accumulating and light‐sensitive chlD–1 mutant displays the expected downregulation of the same nuclear genes. Although chlD–1 and the wild type have similar GUN4 transcript levels, the GUN4 protein in chlD–1 is hardly detectable. Overexpression of GUN4 in chlD–1 modifies the downregulation of nuclear gene expression, but also increases light tolerance. Therefore, GUN4 is proposed to function in ‘shielding’ Proto, and most likely MgProto, by reducing reactivity with O2. Furthermore, GUN4 seems to be involved in sensing elevated levels of these photoreactive tetrapyrrole intermediates, and contributing to 1O2‐mediated retrograde signalling, originating from chlorophyll biosynthesis.
The evolutionary appearance of non‐cyanogenic hydroxynitrile glucosides in the Lotus genus is accompanied by the substrate specialization of paralogous β–glucosidases resulting from a crucial amino acid substitution
Authors: Lai Daniela , Abou Hachem Maher , Robson Fran , Olsen Carl Erik , Wang Trevor L. , Møller Birger L. , Takos Adam M. , Rook Fred 
Published: May 2014

SummaryLotus japonicus, like several other legumes, biosynthesizes the cyanogenic α–hydroxynitrile glucosides lotaustralin and linamarin. Upon tissue disruption these compounds are hydrolysed by a specific β–glucosidase, resulting in the release of hydrogen cyanide. Lotus japonicus also produces the non‐cyanogenic γ‐ and β–hydroxynitrile glucosides rhodiocyanoside A and D using a biosynthetic pathway that branches off from lotaustralin biosynthesis. We previously established that BGD2 is the only β–glucosidase responsible for cyanogenesis in leaves. Here we show that the paralogous BGD4 has the dominant physiological role in rhodiocyanoside degradation. Structural modelling, site‐directed mutagenesis and activity assays establish that a glycine residue (G211) in the aglycone binding site of BGD2 is essential for its ability to hydrolyse the endogenous cyanogenic glucosides. The corresponding valine (V211) in BGD4 narrows the active site pocket, resulting in the exclusion of non‐flat substrates such as lotaustralin and linamarin, but not of the more planar rhodiocyanosides. Rhodiocyanosides and the BGD4 gene only occur in L. japonicus and a few closely related species associated with the Lotus corniculatus clade within the Lotus genus. This suggests the evolutionary scenario that substrate specialization for rhodiocyanosides evolved from a promiscuous activity of a progenitor cyanogenic β–glucosidase, resembling BGD2, and required no more than a single amino acid substitution.
Phosphorylation of photosystem II core proteins prevents undesirable cleavage of D1 and contributes to the fine‐tuned repair of photosystem II
Authors: Kato Yusuke , Sakamoto Wataru 
Published: May 2014

SummaryPhotosystem II (PSII) is a primary target for light‐induced damage in photosynthetic protein complexes. To avoid photoinhibition, chloroplasts have evolved a repair cycle with efficient degradation of the PSII reaction center protein, D1, by the proteases FtsH and Deg. Earlier reports have described that phosphorylated D1 is a poor substrate for proteolysis, suggesting a mechanistic role for protein phosphorylation in PSII quality control, but its precise role remains elusive. STN8, a protein kinase, plays a central role in this phosphorylation process. To elucidate the relationship between phosphorylation of D1 and the protease function we assessed in this study the involvement of STN8, using Arabidopsis thaliana mutants lacking FtsH2 [yellow variegated2 (var2)] and Deg5/Deg8 (deg5 deg8). In support of our presumption we found that phosphorylation of D1 increased more in var2. Furthermore, the coexistence of var2 and stn8 was shown to recover the delay in degradation of D1, resulting in mitigation of the high vulnerability to photoinhibition of var2. Partial D1 cleavage fragments that depended on Deg proteases tended to increase, with concomitant accumulation of reactive oxygen species in the mutants lacking STN8. We inferred that the accelerated degradation of D1 in var2 stn8 presents a tradeoff in that it improved the repair of PSII but simultaneously enhanced oxidative stress. Together, these results suggest that PSII core phosphorylation prevents undesirable cleavage of D1 by Deg proteases, which causes cytotoxicity, thereby balancing efficient linear electron flow and photo‐oxidative damage. We propose that PSII core phosphorylation contributes to fine‐tuned degradation of D1.
Fatty acid synthesis and lipid metabolism in the obligate biotrophic fungus Rhizophagus irregularis during mycorrhization of Lotus japonicus
Authors: Wewer Vera , Brands Mathias , Dörmann Peter 
Published: May 2014

SummaryArbuscular mycorrhiza formation with fungi of the Glomeromycota represents a widespread symbiotic interaction of vascular plants. Different signaling events and metabolic adaptations are required for the close interaction between the two partners. Membrane lipid synthesis is a prerequisite for symbiosis, and membrane properties depend on lipid composition. Lipid profiling was performed by liquid chromatography mass spectrometry to study the role of triacylglycerol, diacylglycerol, phospholipids, galactolipids, sterols and sphingolipids during the colonization of Lotus japonicus roots with Rhizophagus irregularis (syn. Glomus intraradices). Mycorrhization leads to an increased phosphate supply and suppresses the increase in galactolipids commonly observed in phosphate‐deprived plants. In addition to free sterols and sterol esters, R. irregularis contains sterol glucosides and acylated sterol glucosides. Glycosylated sphingolipids (glucosylceramide, dihexosylceramide) and inositolphosphorylceramide were detected in the fungus. Lyso‐phosphatidylcholine, a lipid previously implicated in mycorrhiza signaling, is present in low amounts in mock‐infected and mycorrhized roots. The composition of fungal phospholipids changes after mycorrhization because molecular species with palmitvaccenic (di‐16:1) or tetracosenoic (24:1) acyl groups decrease in intraradical mycelium. This adaptation of lipid metabolism during intraradical growth is likely a prerequisite for symbiosis, achieving functional compatibility between the fungal and the periarbuscular membrane. Data mining in genomic and transcript databases revealed the presence of genes encoding enzymes of lipid biosynthesis in R. irregularis. However, no gene encoding multidomain fatty acid de novo synthase was detected in the genome sequence of this obligate biotrophic fungus.
Post‐transcriptional control of GRF transcription factors by microRNA miR396 and GIF co‐activator affects leaf size and longevity
Authors: Debernardi Juan M. , Mecchia Martin A. , Vercruyssen Liesbeth , Smaczniak Cezary , Kaufmann Kerstin , Inze Dirk , Rodriguez Ramiro E. , Palatnik Javier F. 
Published: May 2014

SummaryThe growth‐regulating factors (GRFs) are plant‐specific transcription factors. They form complexes with GRF‐interacting factors (GIFs), a small family of transcriptional co‐activators. In Arabidopsis thaliana, seven out of the nine GRFs are controlled by microRNA miR396. Analysis of Arabidopsis plants carrying a GRF3 allele insensitive to miR396 revealed a strong boost in the number of cells in leaves, which was further enhanced synergistically by an additional increase of GIF1 levels. Genetic experiments revealed that GRF3 can still increase cell number in gif1 mutants, albeit to a much lesser extent. Genome‐wide transcript profiling indicated that the simultaneous increase of GRF3 and GIF1 levels causes additional effects in gene expression compared to either of the transgenes alone. We observed that GIF1 interacts in vivo with GRF3, as well as with chromatin‐remodeling complexes, providing a mechanistic explanation for the synergistic activities of a GRF3–GIF1 complex. Interestingly, we found that, in addition to the leaf size, the GRF system also affects the organ longevity. Genetic and molecular analysis revealed that the functions of GRFs in leaf growth and senescence can be uncoupled, demonstrating that the miR396‐GRF‐GIF network impinges on different stages of leaf development. Our results integrate the post‐transcriptional control of the GRF transcription factors with the progression of leaf development.
Investigational screening for Babesia microti in a large repository of blood donor samples from nonendemic and endemic areas of the United States
Authors: Moritz Erin D. , Winton Colleen S. , Johnson Stephanie T. , Krysztof David E. , Townsend Rebecca L. , Foster Gregory A. , Devine Patricia , Molloy Philip , Brissette Edward , Berardi Victor P. , Stramer Susan L. 
Published: May 2014

BackgroundBabesia microti, a transfusion‐transmissible intraerythrocytic parasite, is increasing in frequency in the United States with no available FDA‐licensed donor screening assay. We utilized investigational arrayed fluorescence immunoassay (AFIA) and polymerase chain reaction (PCR) to detect B. microti antibodies and DNA in blood donors.Study Design and MethodsAFIA and real‐time PCR were performed on frozen paired EDTA plasma (AFIA) and EDTA whole blood (PCR) samples collected from May to September 2010 to 2011 in nonendemic (Arizona [AZ] and Oklahoma [OK]), moderately endemic (Minnesota [MN] and Wisconsin [WI]), and highly endemic (Connecticut [CT] and Massachusetts [MA]) areas of the United States. AFIA utilized B. microti piroplasm as an antigen substrate; PCR primers and probes targeted the B. microti 18S ribosomal RNA gene. Data from AZ and OK were used to calculate specificity. All AFIA‐ or PCR‐positive or ‐inconclusive donors were deferred, notified, and invited to participate in a follow‐up study involving repeat testing and a demographic and risk‐factor questionnaire. Recipient tracing was performed for any cellular component transfused at index, at subsequent donation, or within the prior 12 months.ResultsTesting of 13,269 paired samples included 4022 from AZ and OK, 4167 from MN and WI, and 5080 from CT and MA. B. microti antibody and/or DNA prevalences were 0.025% (95% confidence interval [CI], 0.00%‐0.14%), 0.12% (95% CI, 0.04%‐0.28%), and 0.75% (95% CI, 0.53%‐1.03%) in the nonendemic, mid‐endemic, and high‐endemic regions, respectively. Specificities were 99.95% (95% CI, 99.82%‐99.99%) at a 1‐in‐64 AFIA cutoff and 99.98% (95% CI, 99.86%‐100.00%) at a 1‐in‐128 cutoff.ConclusionsB. microti prevalence followed expected geographical patterns. Screening was feasible with a performance comparable or superior to other infectious disease blood donor screening assays.
New approach to ‘top‐and‐bottom’ whole blood separation using the multiunit TACSI WB system: quality of blood components
Authors: Lotens A. , Najdovski T. , Cellier N. , Ernotte B. , Lambermont M. , Rapaille A. 
Published: May 2014

Background and ObjectivesTACSI whole blood system is designed to combine primary and secondary processing of six whole blood bags into plasma units, buffy coat and red blood cell concentrates. The aim of this study was to investigate the specifications and in vitro storage parameters of blood components compared with standard centrifugation and separation processing.Materials and MethodsWhole blood bags, collected in CRC kits, were treated on a TACSI whole blood system. They were compared with whole blood bags collected in Composelect kits. In addition to routine quality control analyses, conservation studies were performed on red blood cell concentrates for 42 days and on plasma for 6 months. Platelets pools with five buffy coats were also created, and cellular contamination was evaluated.ResultsRed blood cell concentrates produced from TACSI whole blood met European quality requirements. For white blood cell count, one individual result exceeded 1 × 106 cells/unit. All plasma units fell within specifications for residual cellular contamination and storage parameters. The performances of the TACSI whole blood system allow for the preparation of low volume buffy coats with a recovery of 90% of whole blood platelets. Haemoglobin losses in TACSI BC are smaller, but this did not result in higher haemoglobin content of red cells. These BC are suitable for the production of platelet concentrates.ConclusionFrom these in vitro data, red blood cell concentrates produced using TACSI whole blood are suitable for clinical use with a quality at least equivalent to the control group.