bluuebunny/crossref_metadata_2025_split huggingface dataset:
Detection and expression of the phosphonate transporter gene phnD in marine and freshwater picocyanobacteria
Authors: Ilikchyan Irina N. , McKay R. Michael L. , Zehr Jonathan P. , Dyhrman Sonya T. , Bullerjahn George S. 
Published: February 2009

SummaryWe describe a PCR‐based assay designed to detect expression of the phosphonate assimilation gene phnD from picocyanobacteria. The phnD gene encodes the phosphonate binding protein of the ABC‐type phosphonate transporter, present in many of the picocyanobacterial genome sequences. Detection of phnD expression can indicate a capacity of picoplankton to utilize phosphonates, a refractory form of phosphorus that can represent 25% of the high‐molecular‐weight dissolved organic phosphorus pool in marine systems. Primer sets were designed to specifically amplify phnD sequences from marine and freshwater Synechococcus spp., Prochlorococcus spp. and environmental samples from the ocean and Laurentian Great Lakes. Quantitative RT‐PCR from cultured marine Synechococcus sp. strain WH8102 and freshwater Synechococcus sp. ARC‐21 demonstrated induction of phnD expression in P‐deficient media, suggesting that phn genes are regulated coordinately with genes under phoRB control. Last, RT‐PCR of environmental RNA samples from the Sargasso Sea and Pacific Ocean detected phnD expression from the endemic picocyanobacterial population. Synechococcus spp. phnD expression yielded a depth‐dependent pattern following gradients of P bioavailability. By contrast, the Prochlorococcus spp. primers revealed that in all samples tested, phnD expression was constitutive. The method described herein will allow future studies aimed at understanding the utilization of naturally occurring phoshonates in the ocean as well as monitoring the acquisition of synthetic phosphonate herbicides (e.g. glyphosate) by picocyanobacteria in freshwaters.
Willow wood sap promotes the density‐dependent pathogenesis of Brenneria salicis
Authors: Huvenne Hanneke , Messens Eric , Maes Martine 
Published: February 2009

Summary Brenneria salicis resides in symptomless willow (Salix spp.) and other tree species, but only willow trees develop watermark disease. To understand the conversion of B. salicis into a pathogen, its pathogenicity and differential growth in the various tree species are studied. Brenneria salicis was detected by plating and polymerase chain reaction‐based techniques. Cell wall degradation and quorum sensing (QS) were assayed as possible pathogenicity mechanisms in wood. Differences in B. salicis growth capacities were tested in wood sap of the trees. Watermark diseased willow wood contained high concentrations of B. salicis with QS‐induced cellulase activity. In the fall, wood sap of willow, and not of poplar and alder, promoted high density growth of B. salicis. In situ, B. salicis was the dominant bacterial type in willow wood during the fall and winter period. Willow sustains high densities of B. salicis at the time of leaf shedding. The cellulase in the immobilized wood sap has then a long‐lasting contact with the xylem cell wall. Timing of dormancy and subsequent winter conditions might interfere with sap composition, B. salicis density, activity and survival, and be the reason, at least partly, for the variable occurrence of the disease.
Intracellular fate of Francisella tularensis within arthropod‐derived cells
Authors: Santic Marina , Akimana Christine , Asare Rexford , Kouokam Joseph C. , Atay Safinur , Kwaik Yousef Abu 
Published: February 2009

SummarySince transmission of Francisella tularensis into the mammalian host occurs via arthropod vectors such as ticks, mosquitoes, horseflies and deerflies, recent studies have established Drosophila melanogaster as an arthropod vector model system. Nothing is known about the intracellular fate of F. tularensis within arthropod‐derived cells, and the role of this host‐parasite adaptation in the evolution of this pathogen to infect mammals. In this report, we explored intracellular trafficking of F. tularensis ssp. novicida in D. melanogaster‐derived S2 cells. First, we show that similar to the F. tularensis ssp. holarctica‐derived LVS strain, F. tularensis ssp. novicida is highly infectious, replicates exponentially within S2 cells and within adult flies, and is fatal to adult fruit flies in a dose‐dependent manner, while the iglC, iglD and mglA mutants are defective. Using electron and fluorescence microscopy‐based phagosome integrity assays, we show that the wild‐type strain escapes into the cytosol of S2 cells within 30–60 min post infection and by 6 h, 90% were cytosolic. In contrast, approximately 40–50% of the iglC and iglD mutants escape into the cytosol by 6 h while the other subpopulation becomes enclosed within multilamellar vesicles (MLVs). Pre‐treatment of S2 cells with the autophagy inhibitor methyl adenine blocks formation of the MLVs and all the vacuolar subpopulation of the iglC and iglD mutant bacteria become enclosed within single membrane‐surrounded vacuoles. Endocytic trafficking studies of F. tularensis within S2 cells show transient colocalization of the bacterial phagosome with D. melanogaster LAMP2–GFP fusion but not with lysosomes pre‐loaded with fluorescent dextran. Our data show that MLVs harbouring the iglC mutant acquire Lamp2 and dextran while MLVs harbouring the iglD mutant exclude these late endosomal and lysosomal markers. Our data indicate crucial differences in the role of the pathogenicity island‐encoded proteins in modulating intracellular trafficking within human macrophages and arthropod vector‐derived cells.
Biodiversity of arbuscular mycorrhizal fungi in roots and soils of two salt marshes
Authors: Wilde Petra , Manal Astrid , Stodden Marc , Sieverding Ewald , Hildebrandt Ulrich , Bothe Hermann 
Published: February 2009

SummaryThe occurrence of arbuscular mycorrhizal fungi (AMF) was assessed by both morphological and molecular criteria in two salt marshes: (i) a NaCl site of the island Terschelling, Atlantic Coast, the Netherlands and (ii) a K2CO3 marsh at Schreyahn, Northern Germany. The overall biodiversity of AMF, based on sequence analysis, was comparably low in roots at both sites. However, the morphological spore analyses from soil samples of both sites exhibited a higher AMF biodiversity. Glomus geosporum was the only fungus of the Glomerales that was detected both as spores in soil samples and in roots of the AMF‐colonized salt plants Aster tripolium and Puccinellia sp. at both saline sites and on all sampling dates (one exception). In roots, sequences of Glomus intraradices prevailed, but this fungus could not be identified unambiguously from DNA of soil spores. Likewise, Glomus sp. uncultured, only deposited as sequence in the database, was widely detected by DNA sequencing in root samples. All attempts to obtain the corresponding sequences from spores isolated from soil samples failed consistently. A small sized Archaeospora sp. was detected, either/or by morphological and molecular analyses, in roots or soil spores, in dead AMF spores or orobatid mites. The study noted inconsistencies between morphological characterization and identification by DNA sequencing of the 5.8S rDNA‐ITS2 region or part of the 18S rDNA gene. The distribution of AMF unlikely followed the salt gradient at both sites, in contrast to the zone formation of plant species. Zygotes of the alga Vaucheria erythrospora (Xanthophyceae) were retrieved and should not be misidentified with AMF spores.
Dual epidermal growth factor receptor and vascular endothelial growth factor receptor inhibition with vandetanib sensitizes bladder cancer cells to cisplatin in a dose‐ and sequence‐dependent manner
Authors: Flaig Thomas W. , Su Lih‐Jen , McCoach Caroline , Li Yuan , Raben David , Varella‐Garcia Marileila , Bemis Lynne T. 
Published: February 2009

OBJECTIVETo investigate the activity of the combination of vandetanib and cytotoxic agents using in vitro models of bladder cancer, as modern chemotherapy regimens are built around cisplatin, with gemcitabine or a taxane such as docetaxel also commonly added in combination for the treatment of advanced bladder cancer.MATERIALS AND METHODSHuman bladder cancer cells HTB3, HT1376, J82, RT4, CRL1749, T24, SUP and HTB9 were cultured. The activity of gefitinib (ZD1839) and vandetanib (ZD6474) was assessed in these eight bladder cancer cell lines with a tetrazolium‐based assay of cell viability. RT4 bladder cancer cells, determined to have moderate cisplatin resistance and also moderate sensitivity to vandetanib, were treated with vandetanib and cisplatin. RT4 and T24 cells were treated with six different regimens. The apoptosis and cell‐cycle analysis were studied by flow cytometry. Expression of p21 and p27 was detected by Western blotting. Fluorescence in situ hybridization (FISH) analysis of epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor 2 was performed for all cell lines.RESULTSAt equal concentrations, vandetanib was a more potent inhibitor of cell viability, compared to gefitinib. At vandetanib concentrations of ≤2 µM, the combination with cisplatin was synergistic, especially in the treatment sequence of cisplatin followed by vandetanib, and additive with vandetanib followed by cisplatin. An analysis of the cell‐cycle distribution showed that vandetanib treatment induced G1 arrest at high concentrations, but not at lower concentrations. High‐concentration treatment was associated with increased levels of the cyclin‐dependent kinase p27. FISH analysis showed that there was a low level of genomic gain, and no gene amplification. Mutational analysis of exons 18, 19, and 21 of EGFR in each cell line revealed no mutation.CONCLUSIONVandetanib has synergistic activity when given at low concentration with cytotoxic chemotherapy. The addition of vandetanib to cisplatin‐based chemotherapy regimens merits further study.
Patients’ perspective of botulinum toxin‐A as a long‐term treatment option for neurogenic detrusor overactivity secondary to spinal cord injury
Authors: Hori Satoshi , Patki Prasad , Attar Kaka H. , Ismail Soran , Vasconcelos Joana C. , Shah P. Julian R. 
Published: February 2009

OBJECTIVETo evaluate patients’ perspective on whether they would consider botulinum toxin‐A (BTX‐A) injections as a long‐term treatment option for managing their neurogenic detrusor overactivity (NDO) secondary to spinal cord injury (SCI).PATIENTS AND METHODSIn all, 72 patients with SCI and urodynamically confirmed NDO refractory to anticholinergics, who have had at least one or more injections with BTX‐A were invited to participate in a 5‐min telephone questionnaire covering various aspects of their treatment. Questions about patient satisfaction were rated on a scale from 1 to 10 (1, not satisfied; to 10, very satisfied).RESULTSOf the 72 patients surveyed, 48 (67%) were still actively undergoing repeat BTX‐A injections. The mean patient satisfaction score was 6.2. Of the 48 patients, 43 (90%) replied that they would consider continuing with BTX‐A injections as a long‐term treatment option. Only seven (15%) of patients still having BTX‐A injections would consider an alternative permanent surgical option in the next 5 years. Of those patients considering a one‐off permanent surgical solution, younger patients were likely to consider this at a later interval than those in an older group (Spearman’s correlation coefficient, −0.52, 95% confidence interval −0.78 to −0.10, P = 0.02). The annual new patient recruitment rate was high (mean 14.4) and the annual withdrawal rate was low (mean 4.8).CONCLUSIONWith high satisfaction and low annual withdrawal rates, there are increasingly many patients on BTX‐A. Most consider continuing BTX‐A injections in the long term, increasing the future demand for this service. There is an urgent need for further research into optimizing the current delivery of an intradetrusor BTX‐A injection service for patients with NDO.
Superoxide from NADPH oxidase as second messenger for the expression of osteopontin and monocyte chemoattractant protein‐1 in renal epithelial cells exposed to calcium oxalate crystals
Authors: Umekawa Tohru , Tsuji Hidenori , Uemura Hirotsugu , Khan Saeed R. 
Published: February 2009

OBJECTIVETo test the hypothesis that exposure of a renal epithelial cell line, NRK52E, to calcium oxalate monohydrate crystals (COM) would up‐regulate NADPH oxidase subunit p47phox, enhance superoxide production and increase monocyte chemoattractant protein‐1 (MCP‐1) and osteopontin mRNA levels.MATERIALS AND METHODSConfluent cultures of NRK52E cells were exposed to COM (66.7 µg/cm2) with or with no pretreatment with diphenileneiodium chloride (DPI, 10 × 10−6m) an inhibitor for NADPH oxidase, under serum‐free conditions. The conditioned medium was collected and total cellular RNA isolated from the cells, and subjected to enzyme‐linked immunosorbent assay and real‐time polymerase chain reaction (PCR). Production of reactive oxygen species (ROS) was estimated by dihydroethidium (DHE) staining using a fluorescence microscope. Immunohistochemistry and real‐time PCR were used to analyse p47phox in NRK52E cells.RESULTSIn COM treated NRK52E cells there was enhanced expression of p47phox and production of superoxide. COM‐induced production of MCP‐1 and osteopontin was significantly reduced after treatment with DPI.CONCLUSIONSWhile the generation of a lot of ROS might play a major role in tissue injury or death, the regulated generation of low concentration of ROS, possibly by NADPH oxidase, may represent a second messenger system for generation of COM‐induced MCP‐1 and osteopontin production in the renal tubules.
Sleep duration is an independent factor in nocturia: analysis of bladder diaries
Authors: Udo Yukihiro , Nakao Masahiro , Honjo Hisashi , Ukimura Osamu , Kitakoji Hiroshi , Miki Tsuneharu 
Published: February 2009

OBJECTIVETo analyse the relationship between the number of nocturia episodes and other variables obtained from a bladder diary, with special attention to the duration of sleep, as nocturia is closely related to sleep disorders.PATIENTS, SUBJECTS AND METHODSBetween April 2005 and December 2006, 532 participants, including both community‐dwelling subjects who underwent a mass screening programme and outpatients who consulted the department of urology, completed a 3‐day bladder diary. Clinical variables, i.e. the number of daytime and night‐time voids, diurnal and nocturnal urine volume, daytime and night‐time maximum voided volume, sleep duration and nocturnal polyuria index (NPi) were obtained from the diary. The variables were assessed by univariate analysis in all participants, and by multiple regression analysis of all participants, males, females, subjects in mass screening and those with no 24‐h polyuria, to determine which variables were independently associated with the number of nocturia episodes.RESULTSUnivariate analysis showed a considerable correlation between the nocturia episodes and age, nocturnal urine volume, NPi and sleep duration. On multiple regression analysis, age, nocturnal urine volume, night‐time maximum voided volume and sleep duration were independently related to number of nocturia episodes in all subgroups.CONCLUSIONSSleep duration was confirmed as an independent factor in nocturia. Sleep conditions, including sleep duration, should be considered when evaluating and treating patients with nocturia.
Climate and history explain the species richness peak at mid‐elevation for Schizothorax fishes (Cypriniformes: Cyprinidae) distributed in the Tibetan Plateau and its adjacent regions
Authors: Li Jun , He Qixin , Hua Xia , Zhou Jie , Xu Huidan , Chen Jiakuan , Fu Cuizhang 
Published: February 2009

ABSTRACTAim  We studied elevational species richness patterns of Schizothorax fishes and identified the roles of ecological and evolutionary factors in shaping the patterns of elevational diversity.Location  The Tibetan Plateau and its adjacent regions.Methods  We assembled distribution and altitude data for all Schizothorax species using the literature. We merged ecological and evolutionary approaches to test the relationships between species richness and ecological factors (climate, area, the mid‐domain effect) or evolutionary factors (diversification rates and time of colonization).Results  We found that species richness of Schizothorax fishes peaked at mid‐elevations. Rainfall, area, the mid‐domain effect and diversification rate were weak predictors of the richness pattern. Temperature showed a nonlinear relationship with species richness. Temperature and time of colonization were the most important variables in explaining the elevational diversity pattern.Main conclusion  Our findings indicate that the time‐for‐speciation effect and niche conservatism play important roles in variation of species richness.
Healthy ageing: the role of nutrition and lifestyle – a new British Nutrition Foundation Task Force Report
Authors: Stanner S. , Denny A. 
Published: February 2009

SummaryLife expectancy is increasing around the world, and this is likely to have profound effects on many aspects of society, particularly if these extra years are to be associated with ill health. In the UK, for example, lifespan is increasing at the rate of 2 years for each decade, and over 16% of the population are now aged 65 years or over. Ageing is an integral, natural part of life but the way in which we grow old, our health and functional ability all depend not only on our genetic make‐up, but also on the lifestyle choices we have made over our lifetime. Evidence linking dietary and lifestyle factors throughout the life course with healthy ageing is emerging and has been reviewed in‐depth by a British Nutrition Foundation Task Force comprising of experts in the field and chaired by Professor John Mathers. This article summarises some of the findings of Task Force's Report by discussing the effect of diet and lifestyle on common chronic diseases in later life, namely cardiovascular disease, osteoporosis and arthritis, and on promoting healthy mental ageing and good oral health.
Eggs and dietary cholesterol – dispelling the myth
Authors: Gray J. , Griffin B. 
Published: February 2009

SummaryDietary cholesterol in foods such as eggs has only a small and clinically insignificant effect on blood cholesterol, especially when compared with the much greater effects of dietary saturated fatty acids on blood cholesterol. These facts are now well established and, as a consequence, recommendations from major food and health bodies concerning dietary cholesterol have been relaxed in the UK and elsewhere in recent years. However, in the minds of the public, cholesterol in the diet, specifically from eggs, continues to be viewed with suspicion and that view is still reflected in the advice of some professionals.
Current choices in omega 3 supplementation
Authors: Bailey N. 
Published: February 2009

SummaryOur understanding of the significant role that long‐chain fatty acids play in maintaining health and deterring the progression of disease is rapidly expanding. Much attention has been drawn to the benefits of increasing n‐3 (omega‐3) fatty acids in the diet, either through direct fortification of foodstuffs, or through supplementation. However, it is clear that there is some confusion among the general public as to the benefits of foods fortified with short‐chain n‐3 fatty acids compared with long‐chain fatty acids and, indeed, over the role omega 3 fatty acids play in homeostasis. With the supplement market growing quickly and the general public seemingly increasingly ‘health‐conscious’, emphasis should be focused upon educating the public more thoroughly about the biological roles that these fats play. Modern changes in our diet and the current general consensus that the UK is not an ‘oily fish‐loving’ nation mean that many individuals run the risk of consuming insufficient amounts of the long‐chain n‐3 fatty acids for optimal function of the cardiovascular, immune and inflammatory systems. While supplementation is an easy way of modifying fatty acid intake, the quality of many cheaper products favoured by the public as the ‘value for money’ option are unlikely to offer the same health benefits as those that are molecularly distilled to exclude impurities and maximise concentrations of the active ingredients eicosapentaenoic acid and docosahexaenoic acid. This paper discusses the need for the public to be aware of the variation in the quality and dosage of n‐3 supplements, delineates some of the vegetarian alternatives to fish‐derived products and also discusses the potential role that General Practitioners may play in educating their patients about the possible long‐term benefits of such products.
French‐learning toddlers use gender information on determiners during word recognition
Authors: Van Heugten Marieke , Shi Rushen 
Published: February 2009

Abstract In gender‐marking languages, the gender of the noun determines the form of the preceding article. In this study, we examined whether French‐learning toddlers use gender‐marking information on determiners to recognize words. In a split‐screen preferential looking experiment, 25‐month‐olds were presented with picture pairs that referred to nouns with either the same or different genders. The target word in the auditory instruction was preceded either by the correct or incorrect gender‐marked definite article. Toddlers’ looking times to target shortly after article onset demonstrated that target words were processed most efficiently in different‐gender grammatical trials. While target processing in same‐gender grammatical trials recovered in the subsequent time window, ungrammatical articles continued to affect processing efficiency until much later in the trial. These results indicate that by 25 months of age, French‐learning toddlers use gender information on determiners when comprehending subsequent nouns.
Fourteen‐month‐old infants learn similar‐sounding words
Authors: Yoshida Katherine A. , Fennell Christopher T. , Swingley Daniel , Werker Janet F. 
Published: February 2009

Abstract Can infants, in the very first stages of word learning, use their perceptual sensitivity to the phonetics of speech while learning words? Research to date suggests that infants of 14 months cannot learn two similar‐sounding words unless there is substantial contextual support. The current experiment advances our understanding of this failure by testing whether the source of infants’ difficulty lies in the learning or testing phase. Infants were taught to associate two similar‐sounding words with two different objects, and tested using a visual choice method rather than the standard Switch task. The results reveal that 14‐month‐olds are capable of learning and mapping two similar‐sounding labels; they can apply phonetic detail in new words. The findings are discussed in relation to infants’ concurrent failure, and the developmental transition to success, in the Switch task.