bluuebunny/crossref_metadata_2025 huggingface dataset:
Flight Initiation Distance and Starting Distance: Biological Effect or Mathematical Artefact?
Authors: Dumont François , Pasquaretta Cristian , Réale Denis , Bogliani Giuseppe , von Hardenberg Achaz
Published: August 2012

In many studies, flight initiation distance (FID, the distance at which a prey starts to flee at the approach of a walker) is positively related to starting distance (SD, the distance at which the walker begins to approach) and alert distance (AD, the distance at which the focal individual becomes alert to the threat). In spite of the fundamental differences between SD, a covariate that may not have any biological effect, and AD, a measure related to the behaviour of the animal, it is common to use SD as a proxy for AD when AD is hard to measure (e.g. in species that do not exhibit distinguishable alert postures). However, the relationship between SD and AD or FID may not have any biological reasons, but may instead simply result from a mathematical artefact because of the constraints SD ≥ AD ≥ FID. Under such constrains, the homoscedasticity assumption is violated, and thus, the classical null hypothesis of linear regression (slope = 0) is invalid. In this study, we first show that using SD as a proxy for AD can strongly affect the results on FID. Using data from FID tests on alpine marmots (Marmota marmota), a linear mixed model with AD as a covariate, suggested that the interaction between previous activity and AD had an effect on FID, while this effect was not detected when SD replaced AD as the covariate in the analysis. We then propose that the actual statistical test of the relationship between SD, AD and FID should be based on a null hypothesis that incorporates the constraint SD ≥ AD ≥ FID ≥ 0 and generate 95% CI of simulated slopes obtained from random values under this constraint. This null hypothesis can be rejected if the observed slope of the relationship between two of these variables is outside the 95% CI. We demonstrated that, for alpine marmots, the observed slope of the relationship between AD and SD was within the 95% CI of the simulated slopes. The absence of a statistically significant biological effect in the relationship between SD and AD raises important questions on the outcome of relationship between SD and FID. In Alpine marmot flight, decision should be studied separating the effect of SD on AD and the effect of AD on FID.
Expressions and inhibitory functions of CD300a receptors on purified human basophils
Authors: Gibbs Bernhard F. , Sabato Vito , Bridts Chris H. , Ebo Didier G. , Ben‐Zimra Micha , Levi‐Schaffer Francesca
Published: August 2012

The inhibitory myeloid immunoglobulin receptor CD300a (IRp60) has been shown to downregulate mast cell and eosinophil activities, thereby serving as a potential target for inhibiting allergic effector cell input in allergy. Our aims were to study the expression and functional properties of this receptor in purified human basophils, cells that crucially contribute to Th2‐type immunity and allergy. Basophils homogeneously expressed CD300a as well as the inhibitory receptor CD200R on their cell surface, and these expressions increased after anti‐IgE stimulation. IgE‐mediated basophil degranulation was also significantly inhibited by crosslinking of either CD200R or CD300a (by 90% and 50%, respectively). Inhibitory SHIP‐1 phosphorylations were also induced by CD200R and CD300a, although they were not noticeably increased by IgE‐dependent activation. We conclude that both CD200R and CD300a play a role in reducing IgE‐mediated basophil function and may crucially govern the known differential activities of these cells in vivo.
In vitro formation of organized structure between keratinocytes and dorsal‐root‐ganglion cells
Authors: Tsutsumi Moe , Nakatani Masashi , Kumamoto Junichi , Denda Sumiko , Denda Mitsuhiro
Published: August 2012

We recently found that the morphology in a co‐culture system of keratinocytes and dorsal‐root ganglion‐derived cells depended on the timing of seeding of the two cell types. In skin, epidermis is formed first, followed by construction of peripheral nerve structure. Therefore, we hypothesized that formation of peripheral nerve structure in the epidermis might be driven by interaction between keratinocytes and nerve cells. In the present study, we tested this idea by incubating keratinocytes and dorsal‐root ganglion cells in a spatially separated manner and observing the morphological changes in the co‐culture system. Extension of nerve fibre‐like structures from the ganglion cells was observed, and within 3 days after seeding, many nerve fibre‐like extensions penetrated into the keratinocyte cluster, subsequently forming a network that appeared to resemble the cutaneous peripheral nervous system. Our present model may be useful for studying the formation of peripheral nerve structure in the skin.
Long‐term evolutionary and ecological responses of calcifying phytoplankton to changes in atmospheric CO2
Authors: Hannisdal Bjarte , Henderiks Jorijntje , Liow Lee Hsiang
Published: August 2012

Calcifying phytoplankton play an important role in marine ecosystems and global biogeochemical cycles, affecting the transfer of both organic and inorganic carbon from the surface to the deep ocean. Coccolithophores are the most prominent members of this group, being well adapted to low‐nutrients environments (e.g., subtropical gyres). Despite urgent concerns, their response to rising atmospheric carbon dioxide levels (pCO2) and ocean acidification is still poorly understood, and short‐term experiments may not extrapolate into longer‐term climatic adaptation. Current atmospheric pCO2 (~390 ppmv) is unprecedented since at least 3 million years ago (Ma), and levels projected for the next century were last seen more than 34 Ma. Hence, a deep‐time perspective is needed to understand the long‐term effects of high pCO2 on the biosphere. Here we combine a comprehensive fossil data set on coccolithophore cell size with a novel measure of ecological prominence: Summed Common Species Occurrence Rate (SCOR). The SCOR is decoupled from species richness, and captures changes in the extent to which coccolithophores were common and widespread, based on global occurrences in deep‐sea sediments. The size and SCOR records are compared to state‐of‐the‐art data on climatic and environmental changes from 50 to 5 Ma. We advance beyond simple correlations and trends to quantify the relative strength and directionality of information transfer among these records. Coccolithophores were globally more common and widespread, larger, and more heavily calcified in the pre‐34 Ma greenhouse world, and declined along with pCO2 during the Oligocene (34–23 Ma). Our results suggest that atmospheric pCO2 has exerted an important long‐term control on coccolithophores, directly through its availability for photosynthesis or indirectly via weathering supply of resources for growth and calcification.
Human phagosome processing of Mycobacterium tuberculosis antigens is modulated by interferon‐γ and interleukin‐10
Authors: Bobadilla Karen , Sada Eduardo , Jaime Maria E. , González Yolanda , Ramachandra Lakshmi , Rojas Roxana E. , Pedraza‐Sánchez Sigifredo , Michalak Colette , González‐Noriega Alfonso , Torres Martha
Published: August 2012

Intracellular pathogens, such as Mycobacterium tuberculosis, reside in the phagosomes of macrophages where antigenic processing is initiated. Mycobacterial antigen–MHC class II complexes are formed within the phagosome and are then trafficked to the cell surface. Interferon‐γ (IFN‐γ) and interleukin‐10 (IL‐10) influence the outcome of M. tuberculosis infection; however, the role of these cytokines with regard to the formation of M. tuberculosis peptide–MHC‐II complexes remains unknown. We analysed the kinetics and subcellular localization of M. tuberculosis peptide–MHC‐II complexes in M. tuberculosis‐infected human monocyte‐derived macrophages (MDMs) using autologous M. tuberculosis‐specific CD4+ T cells. The MDMs were pre‐treated with either IFN‐γ or IL‐10 and infected with M. tuberculosis. Cells were mechanically homogenized, separated on Percoll density gradients and manually fractionated. The fractions were incubated with autologous M.  tuberculosis ‐specific CD4+ T cells. Our results demonstrated that in MDMs pre‐treated with IFN‐γ, M. tuberculosis peptide–MHC‐II complexes were detected early mainly in the phagosomal fractions, whereas in the absence of IFN‐γ, the complexes were detected in the endosomal fractions. In MDMs pre‐treated with IL‐10, the M. tuberculosis peptide–MHC‐II complexes were retained in the endosomal fractions, and these complexes were not detected in the plasma membrane fractions. The results of immunofluorescence microscopy demonstrated the presence of Ag85B associated with HLA‐DR at the cell surface only in the IFN‐γ‐treated MDMs, suggesting that IFN‐γ may accelerate M. tuberculosis antigen processing and presentation at the cell membrane, whereas IL‐10 favours the trafficking of Ag85B to vesicles that do not contain LAMP‐1. Therefore, IFN‐γ and IL‐10 play a role in the formation and trafficking of M. tuberculosis peptide–MHC‐II complexes.
A randomized trial of the safety, pharmacokinetics and pharmacodynamics of edoxaban, an oral factor Xa inhibitor, following a switch from warfarin
Authors: Mendell Jeanne , Noveck Robert J. , Shi Minggao
Published: August 2012

Aims

To evaluate the safety, tolerability, pharmacokinetics and pharmacodynamics of edoxaban, an oral direct factor Xa inhibitor, in healthy subjects switching from warfarin.

Methods

Seventy‐two subjects were randomized to edoxaban 60 mg once daily (n = 48) or matching placebo (n = 24) for 5 days at 24 h after the last dose of warfarin treatment (INR 2.0 to 3.0). Safety/tolerability was the primary outcome measure. Pharmacokinetics, INR, aPTT, anti‐FXa, thrombin generation and other coagulation assays were assessed.

Results

Seventy‐two subjects were randomized and 64 subjects received at least one dose of edoxaban (n = 43) or placebo (n = 21) after achieving a target INR of 2.0 to 3.0 on warfarin treatment. Edoxaban 60 mg administered 24 h post‐warfarin appeared to be safe and well tolerated. Adverse events were similar across treatments. For bleeding‐related adverse events, eight subjects tested positive for faecal occult blood, five subjects during warfarin treatment and three subjects during edoxaban treatment. The mean (SD) baseline (post‐dose of warfarin) INR was 2.31 (0.193) which increased to 3.84 (0.744) over 2 h during the edoxaban treatment (P < 0.0001 vs. placebo), returning to post‐warfarin baseline within 12 h. A similar time course of effects for the other coagulation assays was observed in accordance with the drugs' mechanisms of action.

Conclusion

In this study of healthy subjects, edoxaban administered 24 h after the last dose of warfarin was safe and well tolerated with transient increases across the various coagulation assays above post‐warfarin baseline levels.
Higher anticholinergic drug scale (ADS) scores are associated with peripheral but not cognitive markers of cholinergic blockade. Cross sectional data from 21 Norwegian nursing homes
Authors: Kersten Hege , Molden Espen , Willumsen Tiril , Engedal Knut , Wyller Torgeir Bruun
Published: August 2012

Aim

This study evaluated a presumed gradual decline in cognitive function in nursing home residents when the anticholinergic drug scale (ADS) score increased above 3.

Method

The study population was recruited from 21 nursing homes in Norway. Criteria for inclusion were ADS score ≥ 3 and no severe dementia, defined as Clinical Dementia Rating (CDR) score < 3. Primary cognitive end points were CERAD 10‐word lists for recall and Mini Mental State Examination (MMSE). Secondary end points were activity of daily living (ADL), mouth dryness and serum anticholinergic activity (SAA). The patients were stratified into subgroups according to ADS score, i.e. a reference group with score 3 and test groups with scores 4, 5 or ≥6. End points were compared by analyses of covariance (ancova).

Results

Overall, 230 of the 1101 screened nursing home residents (21%) had an ADS score ≥3. After exclusion 101 residents were recruited and among these, 87 managed to participate in the study. No significant differences were detected in cognitive function or ADL when ADS increased above 3 (P > 0.10), but in vivo (mouth dryness) and in vitro (SAA) measures of peripheral anticholinergic activity were significantly higher in patients with an ADS score ≥6 (P < 0.01).

Conclusion

The present study does not support a progressive decline in cognitive function with ADS score above 3. This might indicate that the ADS score model has limited potential to predict the clinical risk of central anticholinergic side effects in frail elderly patients receiving multiple anticholinergic drugs.
Comparison of cell proliferation and epigenetic modification of gene expression patterns in canine foetal fibroblasts and adipose tissue‐derived mesenchymal stem cells
Authors: Oh H. J. , Park E. J. , Lee S. Y. , Soh J. W. , Kong I. S. , Choi S. W. , Ra J. C. , Kang S. K. , Lee B. C.
Published: August 2012

Objectives

This study compared rate of cell proliferation, viability, cell size, expression patterns of genes related to pluripotency and epigenetic modification between canine foetal fibroblasts (cFF) and canine adipose tissue‐derived mesenchymal stem cells (cAd‐MSC).

Materials and methods

Proliferation pattern, cell viability as well as cell size at each passage of cFF and cAd‐MSC were measured when cultures reached confluence. In addition, real‐time PCR was performed to investigate expression of Dnmt1, HDAC1, OCT4, SOX2, BAX, BCL2 genes with reference to β‐actin gene expression as an endogenous control in both cell lines.

Results

cFF and cAd‐MSC differed in number of generations, but not in doubling times, at all passages. Mean cell size of cAd‐MSC was significantly smaller than that of cFF. Cell viability was significantly lower in cFFs and apoptotic level was significantly lower in cAd‐MSC compared to passage‐matched cFF. In the expression of genes related to pluripotency and epigenetic modification, level of HDAC1 in cAd‐MSC was significantly higher than in cFF, but expression of Dnmt1 did not differ between the two groups. OCT4 and SOX2 were significantly more highly expressed in cAd‐MSC compared to cFF.

Conclusions

cAd‐MSC have higher stem‐cell potential than cFF in terms of proliferation patterns, epigenetic modification and pluripotency, thus cAd‐MSC could be more appropriate than cFF as donors of nuclei in somatic cell nuclear transfer for transgenesis.
Efficacy and immunomodulatory actions of ONO-4641, a novel selective agonist for sphingosine 1-phosphate receptors 1 and 5, in preclinical models of multiple sclerosis
Authors: Komiya T , Sato K , Shioya H , Inagaki Y , Hagiya H , Kozaki R , Imai M , Takada Y , Maeda T , Kurata H , Kurono M , Suzuki R , Otsuki K , Habashita H , Nakade S
Published: August 2012

ONO-4641 is a next-generation sphingosine 1-phosphate (S1P) receptor agonist selective for S1P receptors 1 and 5. The objective of the study was to characterize the immunomodulatory effects of ONO-4641 using preclinical data. ONO-4641 was tested in both in-vitro pharmacological studies as well as in-vivo models of transient or relapsing–remitting experimental autoimmune encephalomyelitis (EAE). In vitro, ONO-4641 showed highly potent agonistic activities versus S1P receptors 1 and 5 [half maximal effective concentration (EC50) values of 0·0273 and 0·334 nM, respectively], and had profound S1P receptor 1 down-regulating effects on the cell membrane. ONO-4641 decreased peripheral blood lymphocyte counts in rats by inhibiting lymphocyte egress from secondary lymphoid tissues. In a rat experimental autoimmune encephalomyelitis (EAE) model, ONO-4641 suppressed the onset of disease and inhibited lymphocyte infiltration into the spinal cord in a dose-dependent manner at doses of 0·03 and 0·1 mg/kg. Furthermore, ONO-4641 prevented relapse of disease in a non-obese diabetic mouse model of relapsing-remitting EAE. These observations suggest that ONO-4641 may provide therapeutic benefits in the treatment of multiple sclerosis.
To be or not to be faithful: flexible fidelity to foraging trails in the leaf‐cutting antAcromyrmex lobicornis
Authors: ELIZALDE LUCIANA , FARJI‐BRENER ALEJANDRO
Published: August 2012

1. Ants using trails to forage have to select between two alternative routes at bifurcations, using two, potentially conflicting, sources of information to make their decision: individual experience to return to a previous successful foraging site (i.e. fidelity) and ant traffic. In the field, we investigated which of these two types of information individuals of the leaf‐cutting antAcromyrmex lobicornisEmery use to decide which foraging route to take.

2. We measured the proportion of foraging ants returning to each trail of bifurcations the following day, and for 4–7 consecutive days. We then experimentally increased ant traffic on one trail of the bifurcation by adding additional food sources to examine the effect of increased ant traffic on the decision that ants make.

3. Binomial tests showed that for 62% of the trails, ant fidelity was relatively more important than ant traffic in deciding which bifurcation to follow, suggesting the importance of previous experience.

4. When information conflict was generated by experimentally increasing ant traffic along the trail with less foraging activity, most ants relied on ant traffic to decide. However, in 33% of these bifurcations, ants were still faithful to their trail. Thus, there is some degree of flexibility in the decisions thatA. lobicornismake to access food resources.

5. This flexible fidelity results in individual variation in the response of workers to different levels of ant traffic, and allows the colony to simultaneously exploit both established and recently discovered food patches, aiding efficient food gathering.
Asymmetric priority effects influence the success of invasive forest insects
Authors: MILLER‐PIERCE MAILEA R. , PREISSER EVAN L.
Published: August 2012

1. Settlement timing is often an important factor in interspecific herbivore interactions, as early‐arriving species may encounter higher resource availability and/or avoid induced defences. Despite the general importance of priority effects to the outcome of herbivore interactions, there has been little exploration of such interactions on woody host plants where their impact can only be measured over multiple years.

2. In the eastern U.S.A., two invasive species, the hemlock woolly adelgid Adelges tsugae and the elongate hemlock scale Fiorinia externa, share a native host, eastern hemlock Tsuga canadensis. Their interaction and its consequences were investigated for plant growth – hemlock saplings that had been inoculated with either A. tsugae or F. externa, starting in spring 2007, were cross‐infested with the other insect in spring 2009. A set of uninfested trees was simultaneously infested with A. tsugae, F. externa, both, or neither insect (= control), and insect density and plant growth was assessed in all treatments.

3. Adelges tsugae settlement rates did not differ if it settled alone or simultaneously with F. externa, but were ∼45% lower on trees previously infested with F. externa. There was no difference in F. externa settlement rates, and plant growth did not differ substantively between any of the herbivore treatments.

4. At a temporal scale (i.e. multiple growing seasons) appropriate to interactions between woody plants and their herbivores, this work demonstrates that plant‐mediated priority effects can substantially affect herbivore settlement and thus the outcome of interspecific competition.
Acute lung injury results from failure of neutrophil de‐priming: a new hypothesis
Authors: Singh Nanak R. P. , Johnson Andrew , Peters A. Michael , Babar Judith , Chilvers Edwin R. , Summers Charlotte
Published: August 2012

Eur J Clin Invest 2012; 42 (12): 1342–1349

Neutrophils are the most abundant circulating white cell in humans and play a crucial role in the innate immune response. Accumulation and activation of neutrophils, together with delayed clearance, have been shown to be a key event in the pathogenesis of acute lung injury. Previously, it has been proposed that there is substantial pooling of neutrophils within the pulmonary vasculature, even under physiological conditions, making the lung especially vulnerable to neutrophil‐mediated tissue injury. However, more recent evidence suggests that only primed neutrophils accumulate in the pulmonary vasculature. This article examines the evidence for these two opposing views and proposes a new two‐step model for the recruitment of neutrophils into the lung. Firstly, neutrophils that become primed, by exposure to a range of inflammatory mediators or physicochemical perturbations, become shape changed and stiff because of alterations in their cytoskeleton, and as a result, accumulate within the pulmonary circulation. In the absence of further stimuli, the healthy pulmonary vasculature is able to selectively retained these primed cells, allow them to ‘de‐prime’ and be released back into the circulation in a quiescent, state. If this pulmonary ‘de‐priming’ mechanism fails, or a second insult occurs, such as ventilator‐associated barotrauma, which causes loss of alveolar integrity, primed neutrophils migrate from the pulmonary vasculature into the interstitial space with resultant lung injury. This canonical ‘two step’ model highlights the importance of neutrophil priming in the genesis of lung injury and the importance of adopting strategies to minimise alveolar injury.
Comprehensive care of the patient with haemophilia and inhibitors undergoing surgery: practical aspects
Authors: Kulkarni R.
Published: August 2012

Congenital haemophilia is a rare and complex condition for which dedicated specialized and comprehensive care has produced measurable improvements in clinical outcomes and advances in patient management. Among these advances is the ability to safely perform surgery in patients with inhibitor antibodies to factors VIII and IX, in whom all but the most necessary of surgeries were once avoided due to the risk for uncontrollable bleeding due to ineffectiveness of replacement therapy. Nevertheless, surgery continues to pose a major challenge in this relatively rare group of patients because of significantly higher costs than in patients without inhibitors, as well as a high risk for bleeding and other complications. Because of the concentration of expertise and experience, it is recommended that any surgery in patients with haemophilia and inhibitors be planned in conjunction with a haemophilia treatment centre (HTC) and performed in a hospital that incorporates a HTC. Coordinated, standard pre‐, intra‐ and postoperative assessments and planning are intended to optimize surgical outcome and utilization of resources, including costly factor concentrates and other haemostatic agents, while minimizing the risk for bleeding and other adverse consequences both during and after surgery. This article will review the special considerations for patients with inhibitors as they prepare for and move through surgery and recovery, with an emphasis on the roles and responsibilities of individual members of the multidisciplinary team in facilitating this process.
Detection of tick‐borne pathogens in ticks from migratory birds in the Baltic region of Russia
Authors: MOVILA A. , ALEKSEEV A. N. , DUBININA H. V. , TODERAS I.
Published: August 2012

We report the finding of tick‐borne encephalitis (TBE)‐virus in indigenous Ixodes ricinus (L.), ‘Candidatus Neoehrlichia mikurensis’ in exotic Ixodes frontalis (Panzer) and Rickettsia aeshlimannii in exotic Hyalomma marginatum Koch subadult ticks detached from 18.5% (107/577) infested migratory birds in the Baltic region of Russia. This is the first record of human pathogenic ‘Candidatus N. mikurensis’ in I. frontalis ticks. Moreover, seven other pathogens were identified in I. ricinus ticks. Spotted Fever Group rickettsiae were the predominant pathogen group and were detected only in nymphs. Future investigations are warranted to further characterize the role of birds in the epizootiology of tick‐borne pathogens in this region.
Mitochondrial DNA (COI) analyses reveal that amphipod diversity is associated with environmental heterogeneity in deep‐sea habitats
Authors: Knox Matthew A. , Hogg Ian D. , Pilditch Conrad A. , Lörz Anne‐Nina , Hebert Paul D. N. , Steinke Dirk
Published: August 2012

The relationship between species diversity and environmental parameters is poorly understood for the mobile macrofauna of deep‐sea habitats due to under‐sampling and subsequent lack of accurate taxonomic information. To redress this, cytochrome oxidase c subunit I (COI) DNA sequences were used to estimate species diversity and to compare phoxocephalid amphipod assemblages among 20 stations encompassing a range of environmental conditions. Two regions, east (Chatham Rise) and west (Challenger Plateau) of New Zealand were sampled to depths of 200–1200 m with an epibenthic sled. Using a comparison among identified morphospecies, we found a clear gap in sequence divergences between 6% and 13% and used a 6% threshold to designate molecular operational taxonomic units (MOTUs), as a surrogate to putative species. DNA sequences (n = 297) revealed high total diversity (n = 49 MOTUs), as well as high beta diversity (28 MOTUs found at single location only). Novel phoxocephalid MOTUs were found at most stations, especially on Challenger Plateau and the flanks of Chatham Rise. Analyses of interstation assemblages revealed a major split between regions, indicating minimal overlap in taxon distributions. A cluster of highly similar stations was identified, broadly distributed over the crest of Chatham Rise, in association with elevated food availability, probably resulting from higher surface productivity and relatively shallow depth. Accordingly, multivariate analysis revealed a strong correlation between phoxocephalid assemblages and food supply. This study highlights the value of molecular approaches, in particular COI sequences, for quantifying and comparing diversity in under‐sampled and/or under‐studied taxa.
Genome‐wide patterns of divergence and gene flow across a butterfly radiation
Authors: Nadeau Nicola J. , Martin Simon H. , Kozak Krzysztof M. , Salazar Camilo , Dasmahapatra Kanchon K. , Davey John W. , Baxter Simon W. , Blaxter Mark L. , Mallet James , Jiggins Chris D.
Published: August 2012

The Heliconius butterflies are a diverse recent radiation comprising multiple levels of divergence with ongoing gene flow between species. The recently sequenced genome of Heliconius melpomene allowed us to investigate the genomic evolution of this group using dense RAD marker sequencing. Phylogenetic analysis of 54 individuals robustly supported reciprocal monophyly of H. melpomene and Heliconius cydno and refuted previous phylogenetic hypotheses that H. melpomene may be paraphylectic with respect to H. cydno. Heliconius timareta also formed a monophyletic clade closely related but distinct from H. cydno with Heliconius heurippa falling within this clade. We find evidence for genetic admixture between sympatric populations of the sister clades H. melpomene and H. cydno/timareta, particularly between H. cydno and H. melpomene from Central America and between H. timareta and H. melpomene from the eastern slopes of the Andes. Between races, divergence is primarily explained by isolation by distance and there is no detectable genetic population structure between parapatric races, suggesting that hybrid zones between races are not zones of secondary contact. Our results also support previous findings that colour pattern loci are shared between populations and species with similar colour pattern elements. Furthermore, this pattern is almost unique to these genomic regions, with only a very small number of other loci showing significant similarity between populations and species with similar colour patterns.
The nucleoid‐associated protein Fis directly modulates the synthesis of cellulose, an essential component of pellicle–biofilms in the phytopathogenic bacterium Dickeya dadantii
Authors: Prigent‐Combaret Claire , Zghidi‐Abouzid Ouafa , Effantin Géraldine , Lejeune Philippe , Reverchon Sylvie , Nasser William
Published: August 2012

Bacteria use biofilm structures to colonize surfaces and to survive in hostile conditions, and numerous bacteria produce cellulose as a biofilm matrix polymer. Hence, expression of the bcs operon, responsible for cellulose biosynthesis, must be finely regulated in order to allow bacteria to adopt the proper surface‐associated behaviours. Here we show that in the phytopathogenic bacterium, Dickeya dadantii, production of cellulose is required for pellicle–biofilm formation and resistance to chlorine treatments. Expression of the bcs operon is growth phase‐regulated and is stimulated in biofilms. Furthermore, we unexpectedly found that the nucleoid‐associated protein and global regulator of virulence functions, Fis, directly represses bcs operon expression by interacting with an operator that is absent from the bcs operon of animal pathogenic bacteria and the plant pathogenic bacterium Pectobacterium. Moreover, production of cellulose enhances plant surface colonization by D. dadantii. Overall, these data suggest that cellulose production and biofilm formation may be important factors for surface colonization by D. dadantii and its subsequent survival in hostile environments. This report also presents a new example of how bacteria can modulate the action of a global regulator to co‐ordinate basic metabolism, virulence and modifications of lifestyle.
Tandem attenuators control expression of the Salmonella mgtCBR virulence operon
Authors: Lee Eun‐Jin , Groisman Eduardo A.
Published: August 2012

The mgtCBR operon from Salmonella enterica serovar Typhimurium specifies the virulence protein MgtC, the Mg2+ transporter MgtB and the regulatory peptide MgtR. The mgtCBR transcript includes a long leader region harbouring two short open reading frames (ORFs). Translation of these ORFs is anticipated to impact the formation of particular stem‐loop structures and control transcription of the coding region by an attenuation‐like mechanism. We previously reported that ORF mgtM enables Salmonella to promote transcription of the mgtC and mgtB coding regions when experiencing a rise in cytoplasmic ATP levels. We now show that the proline codon‐rich ORF mgtP mediates an increase in transcription of the mgtC and mgtB coding regions under conditions predicted to decrease the levels of proline‐charged tRNAPro. The high ATP and low proline signals act independently in an additive form. Replacing conserved mgtP proline codons with codons specifying other amino acids abolished the response to proline limitation but had no effect on the response to ATP. Substitution of conserved adenine nucleotides in mgtM abolished the response to ATP but had no effect in the response to proline limitation. This provides a singular example of a leader mRNA with tandem attenuators responding to different signals.
Intranasal Co‐delivery with the Mouse Zona Pellucida 3 and GM‐CSF Expressing Constructs Enhances Humoral Immune Responses and Contraception in Mice
Authors: Ma X. , Li J. , Zhang F.
Published: August 2012

Granulocyte–monocyte colony‐stimulating factor (GM‐CSF) regulates the function of antigen‐presenting cells (APCs) and has been broadly used as the adjuvant. Here, we tested whether intranasal delivery of GM‐CSF can improve the contraception of mouse zona pellucida 3 (mZP3) DNA vaccine. Our results showed that co‐administration of GM‐CSF and mZP3 DNA vaccine increased the levels of secretory IgA (sIgA) and IgG antibodies in vaginal washes and serum, respectively. Co‐administration enhanced Th2 responses through improving the maturation of dendritic cells. Importantly, GM‐CSF significantly reduced the fertility rate and mean litter size induced by mZP3 DNA vaccine alone without interfering the normal follicular development. These data suggest that GM‐CSF could be used as adjuvant to develop immunocontraceptive vaccine.
Gentamicin in Combination with Ascorbic Acid Regulates the severity of Staphylococcus aureus Infection–Induced Septic Arthritis in Mice
Authors: Mal P. , Dutta S. , Bandyopadhyay D. , Dutta K. , Basu A. , Bishayi B.
Published: August 2012

To study the effects of gentamicin in combination with ascorbic acid on septic arthritis, mice were infected with Staphylococcus aureus (S. aureus) and treated with gentamicin, which was given at 5 mg/kg after 24 h of infection, followed by ascorbic acid, given at 20 mg/kg body weight after 2 h of gentamicin treatment. Mice were sacrificed at 3, 9, 15 days post‐infection (dpi). Combined treatment of infected mice with gentamicin and ascorbic acid eradicated the bacteria from the blood, spleen and synovial tissue and showed a significant gross reduction in arthritis, reduced serum levels of tumour necrosis factor alpha (TNF‐α) and interferon gamma (IFN‐γ). S. aureus‐infected mice have demonstrated the disturbed antioxidant status measured in terms of cellular antioxidants like reduced glutathione and antioxidant enzymes such as superoxide dismutase (SOD) and catalase. The same were ameliorated when the animals were co‐treated with gentamicin along with ascorbic acid.
Proteomic analysis of S‐nitrosylated proteins in potato plant
Authors: Kato Hiroaki , Takemoto Daigo , Kawakita Kazuhito
Published: August 2012

Nitric oxide (NO) has various functions in physiological responses in plants, such as development, hormone signaling and defense. The mechanism of how NO regulates physiological responses has not been well understood. Protein S‐nitrosylation, a redox‐related modification of cysteine thiol by NO, is known to be one of the important post‐translational modifications to regulate activity and interactions of proteins. To elucidate NO function in plants, proteomic analysis of S‐nitrosylated proteins in potato (Solanum tuberosum) was performed. Detection and functional analysis of internal S‐nitrosylated proteins is technically demanding because of the instability and reversibility of the protein S‐nitrosylation. By using a modified biotin switch assay optimized for potato tissues, and nano liquid chromatography combined with mass spectrometry, approximately 80 S‐nitrosylated candidate proteins were identified in S‐nitrosoglutathione‐treated potato leaves and tuber extracts. Identified proteins included redox‐related enzymes, defense‐related proteins and metabolic enzymes. Some of identified proteins were synthesized in Escherichia coli, and S‐nitrosylation of recombinant proteins was confirmed in vitro. Dehydroascorbate reductase 1 (DHAR1, EC 1.8.5.1), one of the identified S‐nitrosylated target proteins, showed glutathione‐dependent dehydroascorbate‐reducing activity. Either point mutation in a target cysteine of S‐nitrosylation or treatment with an NO donor, S‐nitroso‐l‐cysteine, significantly reduced the activity of DHAR1, indicating that DHAR1 is negatively regulated by S‐nitrosylation of the cysteine residue essential for the enzymatic activity. These results show that the modified method developed in this study can be used to identify proteins regulated by S‐nitrosylation in potato tissues.
Robotic ultrasound probe for tumor identification in robotic partial nephrectomy: Initial series and outcomes
Authors: Kaczmarek Bartosz F , Sukumar Shyam , Petros Firas , Trinh Quoc‐Dien , Mander Navneet , Chen Roger , Menon Mani , Rogers Craig G
Published: August 2012

Objectives:  Accurate tumor identification during partial nephrectomy is essential for successful tumor control. Intraoperative laparoscopic ultrasonography is useful for tumor localization, but the ultrasound probe is controlled by the assistant rather than the surgeon. We evaluated our initial experience using a robotic ultrasound probe that is controlled by the console surgeon.

Methods:  Partial nephrectomy was carried out in 22 consecutive patients between November 2010 and March 2011. A robotic ultrasound probe under console surgeon control was used in all the cases. All patients had at least 1 year follow up.

Results:  Mean patient age was 59 years and mean tumor size was 2.7 cm. There were six hilar tumors (27%) and 21 (95%) endophytic tumors. Mean R.E.N.A.L. nephrometry score was 6.9 (range 6–9). Mean operative time was 205.7 min and mean warm ischemia time was 17.9 min (range 6–28 min). All patients had negative tumor margins and were free of disease recurrence at a mean follow up of 13 months.

Conclusion:  The use of a robotic ultrasound probe during partial nephrectomy allows the surgeon to optimize tumor identification with maximal autonomy, and to benefit from the precision and articulation of the robotic instrument during this key step of the partial nephrectomy procedure.
Intravesical hyaluronic acid for interstitial cystitis/painful bladder syndrome: A comparative randomized assessment of different regimens
Authors: Lai Ming‐Chih , Kuo Yuh‐Chen , Kuo Hann‐Chorng
Published: August 2012

Objectives:  To compare the clinical effectiveness of different regimens of intravesical hyaluronic acid instillation for patients with interstitial cystitis/painful bladder syndrome.

Methods:  A total of 60 patients (age 16–77 years) diagnosed with interstitial cystitis/painful bladder syndrome were enrolled in this prospective, randomized study. A total of 30 patients were assigned to receive four weekly intravesical instillations of 40 mg of hyaluronic acid followed by five monthly instillations (hyaluronic acid‐9 group). Another 30 patients received 12 intravesical instillations of 40 mg hyaluronic acid every 2 weeks (hyaluronic acid‐12 group). Symptomatic changes after hyaluronic acid treatments were assessed using Interstitial Cystitis Symptom and Problem Indexes, pain visual analog scale, functional bladder capacity, frequency and nocturia in voiding diary, maximum flow rate, voided volume, postvoid residual volume, and Quality of Life Index at 1, 3 and 6 months.

Results:  Of the 60 patients, 59 were evaluable at the end of the study. The Interstitial Cystitis Symptom Index, Interstitial Cystitis Problem Index and total score, pain visual analog scale, functional bladder capacity, maximum flow rate, and Quality of Life Index improved significantly after 6 months in both groups. The frequency and voided volume improved significantly only in the hyaluronic acid‐12 group. However, patients with moderate and marked improvement were clinically similar in both groups. The measured variables did not differ between the two groups over the course of the study.

Conclusion:  No significant difference was noted in the therapeutic effect between two hyaluronic acid instillation regimens for treatment of interstitial cystitis/painful bladder syndrome patients. Both groups showed significant improvement in symptom scores and Quality of Life Index.
Diagnostic usefulness of precise examinations with intraductal ultrasonography, peroral cholangioscopy and laparoscopy of immunoglobulin G4‐related sclerosing cholangitis
Authors: Horiguchi Shigeru , Ikeda Fusao , Shiraha Hidenori , Yamamoto Naoki , Sakakihara Ichiro , Noma Yasuhiro , Tsutsumi Koichiro , Kato Hironari , Hagihara Hiroaki , Yasunaka Tetsuya , Nakamura Shinichiro , Kobashi Haruhiko , Kawamoto Hirofumi , Yamamoto Kazuhide
Published: August 2012

Herein, a case of immunoglobulin G4 (IgG4)‐related sclerosing cholangitis is reported. IgG4 was diagnosed based on observations from peroral cholangioscopy and laparoscopy, and these methods are proposed for definitive and precise diagnosis of this disease. A 76‐year‐old male patient with inguinal Paget's disease had intrahepatic bile duct dilatations detected with computed tomography at his periodic check‐up. Magnetic resonance cholangiography showed stenosis of the upper common bile duct and poststenotic dilatation of left intrahepatic bile ducts. The portal tract and bilateral intrahepatic bile ducts were surrounded by a low‐density area, facing a tumor‐like lesion at segment 2. Cytological examinations of the stenotic and dilated lesions revealed no cellular atypia. Histological examination of the tumor showed normal liver tissue with infiltration of lymphocytes, indicating an inflammatory pseudotumor. Peroral cholangioscopy excluded the possibility of biliary cancer and indicated that the stenotic legion was of submucosal, not mucosal, origin. Laparoscopic observations showed discoloration with wide yellowish‐white lobular markings and wide depressed lesions at segments 2 and 7. Liver histology showed mild cholangitis with infiltration of IgG4‐positive plasma cells around the bile ducts. Serum IgG4 levels were elevated. From these findings, the patient was diagnosed with IgG4‐related sclerosing cholangitis. After treatment with prednisolone, blood liver enzymes and IgG4 rapidly normalized, bile duct dilatations improved, and the hepatic pseudotumor disappeared. The cholangitis did not recur. In this case, biliary cancer was ruled out by observation with peroral cholangioscopy, and the spread of cholangitis in the liver periphery was verified with laparoscopy; this information could not be obtained with other modalities.
Functional variant in methionine synthase reductase decreases the risk of Down syndrome in China
Authors: Wang Shao‐Shuai , Feng Ling , Qiao Fu‐Yuan , Lv Juan‐Juan
Published: August 2012

Aim:  Down syndrome (DS) is the most common genetic cause of human mental retardation and the genes involved in homocysteine/folate metabolism may play important roles in this condition. Methionine synthase reductase (MTRR) is one of the key regulatory enzymes involved in the metabolic pathway of homocysteine. We investigated whether the polymorphism C524T of the MTRR gene is associated with DS.

Method:  A total of 104 mothers of children born with DS and 184 healthy mothers were included. The polymorphisms were detected by polymerase chain reaction and restriction fragment length polymorphism analysis. Plasma folate and total plasma homocysteine (t‐Hcy) concentrations were also measured.

Results:  Significant differences in the distributions of C524T alleles were observed between case and control mothers; a decreased risk of DS was associated with the 524TT genotype (OR = 0.34), CT + TT genotype (OR = 0.60). The mean t‐Hcy value in the case group was higher than the mean value in the control group. t‐Hcy concentrations were lower in TT homozygote than CC homozygote among the cases but not among the controls.

Conclusion:  MTRR C524T polymorphism decreases the risk of DS in the Chinese population.
Hypoxia inducible factor‐1α‐mediated activation of survivin in cervical cancer cells
Authors: Bai Haitao , Ge Shengfang , Lu Jian , Qian Guanxiang , Xu Rang
Published: August 2012

Aim:  Hypoxia, a characteristic of almost all types of solid tumors, has been associated with poor outcome in a number of human malignancies. The aim of this study was to investigate the molecular mechanisms involved in hypoxia‐induced activation of the human survivin gene promoter in cervical HeLa cells.

Material and Methods:  Immunohistochemical staining was used to detect the expression of HIF‐1α and survivin in cervical cancer samples and normal cervical samples. Under normoxic and hypoxic conditions, the expression of hypoxia inducible factor (HIF)‐1α and survivin in cervical cancer HeLa cells was detected by quantitative reverse transcription polymerase chain reaction and Western blotting. Luciferase reporter assays was used to investigate the molecular mechanisms in hypoxia‐induced survivin activation. We also studied the effect of HIF‐1α overexpression on the expression of survivin in cervical cancer HeLa cells.

Results:  Significant HIF‐1α and survivin overexpression is associated with cervical cancer, and HIF‐1α protein expression is strongly correlated with survivin protein expression. In cervical cancer cell line (HeLa), hypoxia upregulated both HIF‐1α and survivin expression. Moreover, luciferase reporter assays using survivin core promoter demonstrated that survivin transcription was activated under hypoxia conditions and was associated with HIF‐1α overexpression. The transcriptional activation of reporter genes in response to hypoxia is independent of potential HIF‐1α‐responsive element, located between −86 and −82 regions. HIF‐1α overexpression significantly activated survivin expression.

Conclusion:  Our results demonstrate that survivin expression is upregulated following the induction of HIF‐1α by hypoxia resulting from tumor formation, possibly leading to tumor progression. These findings have potential implication in developing novel cancer therapy targeting HIF‐1.
Screening for and management of high‐grade cervical intraepithelial neoplasia in Bangladesh: A cross‐sectional study comparing two protocols
Authors: Nessa Ashrafun , Rashid Mohammad Harun Ur , E‐Ferdous Noor , Chowdhury Afroza
Published: August 2012

Aim:  To evaluate the feasibility of the ‘see and treat’ protocol for the management of high‐grade cervical intraepithelial neoplasia (CIN) at a colposcopy clinic in Bangladesh.

Material and Methods:  A cross‐sectional and comparative study was carried out between two periods on 358 colposcopy‐diagnosed high‐grade CIN at the colposcopy clinic of Bangabandhu Sheikh Mujib Medical University (BSMMU). During the first period (January 2005 to June 2008), 181 colposcopy‐diagnosed high‐grade CIN patients had cervical biopsy by punch biopsy forceps and histology‐confirmed high‐grade CIN were treated by loop electrosurgical excision procedure (LEEP). During the second period (July 2008 to December 2009), 177 colposcopy‐diagnosed high‐grade CIN were treated by LEEP at their first visit.

Results:  During the first and second periods, 48 of 87 and 55 of 73 histology‐proven high‐grade CIN cases, respectively, received treatment. Among the study population, 37.3% women who had normal or CIN‐I in histology were treated unnecessarily in the second period. The compliance of treatment improved by 20% and failure to receive treatment fell by 20%; these changes were statistically significant (P = 0.006).

Conclusion:  ‘See and treat’ protocol is a well‐accepted, feasible and useful option for management of high‐grade CIN in Bangladesh. It reduces the number of visits to the clinic and failure to receive treatment.
Functional properties of dopamine neurons and co‐expression of vasoactive intestinal polypeptide in the dorsal raphe nucleus and ventro‐lateral periaqueductal grey
Authors: Dougalis Antonios G. , Matthews Gillian A. C. , Bishop Matthew W. , Brischoux Frédéric , Kobayashi Kazuto , Ungless Mark A.
Published: August 2012

The dorsal raphe nucleus (DRN) and ventrolateral periaqueductal grey (vlPAG) regions contain populations of dopamine neurons, often considered to be a dorsal caudal extension of the A10 group [mostly found in the ventral tegmental area (VTA)]. Recent studies suggest they are involved in promoting wakefulness and mediate some of the antinociceptive and rewarding properties of opiates. However, little is known about their electrophysiological properties. To address this, we used Pitx3‐GFP and tyrosine hydroxylase (TH)‐GFP mice to carry out targeted whole‐cell recordings from this population in acute brain slices. We found that DRN/vlPAG dopamine neurons have characteristics similar to most VTA dopamine neurons, but distinct from dorsal raphe serotonin neurons. They fire broad action potentials at a relatively slow, regular rate, exhibit a hyperpolarization‐activated inward current and delayed repolarization, and show spike‐frequency adaptation in response to prolonged depolarization. In addition, they receive fast excitatory and inhibitory synaptic inputs. Moreover, we found co‐expression of vasoactive intestinal polypeptide in small, periaqueductal dopamine neurons, but generally not in larger, more ventral dopamine neurons.