bluuebunny/crossref_metadata_2025 huggingface dataset:
Gut hormones: emerging role in immune activation and inflammation
Authors: Khan W I , Ghia J E
Published: May 2010

Gut inflammation is characterized by mucosal recruitment of activated cells from both the innate and adaptive immune systems. In addition to immune cells, inflammation in the gut is associated with an alteration in enteric endocrine cells and various biologically active compounds produced by these cells. Although the change in enteric endocrine cells or their products is considered to be important in regulating gut physiology (motility and secretion), it is not clear whether the change plays any role in immune activation and in the regulation of gut inflammation. Due to the strategic location of enteric endocrine cells in gut mucosa, these gut hormones may play an important role in immune activation and promotion of inflammation in the gut. This review addresses the research on the interface between immune and endocrine systems in gastrointestinal (GI) pathophysiology, specifically in the context of two major products of enteric endocrine systems, namely serotonin (5-hydroxytryptamine: 5-HT) and chromogranins (Cgs), in relation to immune activation and generation of inflammation. The studies reviewed in this paper demonstrate that 5-HT activates the immune cells to produce proinflammatory mediators and by manipulating the 5-HT system it is possible to modulate gut inflammation. In the case of Cgs the scenario is more complex, as this hormone has been shown to play both proinflammatory and anti-inflammatory functions. It is also possible that interaction between 5-HT and Cgs may play a role in the modulation of immune and inflammatory responses. In addition to enhancing our understanding of immunoendocrine interaction in the gut, the data generated from the these studies may have implications in understanding the role of gut hormone in the pathogenesis of both GI and non-GI inflammatory diseases which may lead ultimately to improved therapeutic strategies in inflammatory disorders.
T cell response to viral antigens in adults and children with common variable immunodeficiency and specific antibody deficiency
Authors: Haveman L M , Scherrenburg J , Maarschalk-Ellerbroek L J , Hoek P D , Schuurman R , De Jager W , Ellerbroek P M , Prakken B J , Van Baarle D , Van Montfrans J M
Published: May 2010

Several T cell abnormalities have been described in common variable immunodeficiency (CVID), a B cell disorder of mainly unknown origin. A subset of CVID patients suffers from frequent reactivations of herpes viruses. We studied T cell function in CVID [and in a subset of paediatric patients with specific antibody deficiency (SAD)] by measuring T cell proliferation and cytokine production in response to herpes virus-antigens in paediatric CVID patients (n = 9) and paediatric SAD patients (n = 5), in adult CVID patients (n = 14) and in healthy controls. Paediatric CVID patients, but not SAD patients, displayed moderately increased CD8+ T cell proliferation in response to cytomegalovirus, human herpes virus type 6B (HHV6-B) and herpes simplex virus compared to controls. CD8+ T cell responses in adult CVID patients tended to be increased in response to cytomegalovirus and herpes simplex virus. In response to stimulation with herpes virus antigens, the proinflammatory cytokines interleukin (IL)-1β, IL-6, tumour necrosis factor (TNF)-α and interferon inducible protein (IP)-10 were produced. Overall, no major differences were detected in cytokine production upon stimulation between patients and controls, although higher IL-10 and IL-12 production was detected in paediatric patients. In conclusion, cellular immunity against herpes virus antigens appears undisturbed in CVID patients, although defects in subpopulations of CVID patients cannot be excluded.
Human skin and oral mucosal dendritic cells as ‘good guys’ and ‘bad guys’ in allergic immune responses
Authors: Novak N , Gros E , Bieber T , Allam J-P
Published: May 2010

Recent progress achieved by an impressive number of studies focusing upon the ontogenesis and immunobiology of epidermal Langerhans cells (LCs) and other cutaneous dendritic cell (DC) populations as well as DCs at oral mucosal tissue has profoundly revised our understanding of the role of DCs in different tissues and microenvironments. By sensing their environment for microbial signals or allergens and bridging innate and adaptive immunity in a sophisticated manner, subtypes of DCs play a critical role in the maintenance of the immunological homeostasis in the periphery. Thereby, DCs, located directly at the interface to the environment, fulfil opposing tasks as they are key players in both the control and the generation of allergic inflammation. Furthermore, it is under ongoing debate whether DCs attenuate or aggravate allergic inflammation. As a consequence, accumulated knowledge gained in this field within the last few years has provided an excellent basis for innovative therapeutic opportunities which tend to target specifically the multi-faceted properties of DCs at distinct anatomical sites.
2D acoustic‐elastic coupled waveform inversion in the Laplace domain
Authors: Bae Ho Seuk , Shin Changsoo , Cha Young Ho , Choi Yunseok , Min Dong‐Joo
Published: May 2010

Although waveform inversion has been intensively studied in an effort to properly delineate the Earth's structures since the early 1980s, most of the time‐ and frequency‐domain waveform inversion algorithms still have critical limitations in their applications to field data. This may be attributed to the highly non‐linear objective function and the unreliable low‐frequency components. To overcome the weaknesses of conventional waveform inversion algorithms, the acoustic Laplace‐domain waveform inversion has been proposed. The Laplace‐domain waveform inversion has been known to provide a long‐wavelength velocity model even for field data, which may be because it employs the zero‐frequency component of the damped wavefield and a well‐behaved logarithmic objective function. However, its applications have been confined to 2D acoustic media.

We extend the Laplace‐domain waveform inversion algorithm to a 2D acoustic‐elastic coupled medium, which is encountered in marine exploration environments. In 2D acoustic‐elastic coupled media, the Laplace‐domain pressures behave differently from those of 2D acoustic media, although the overall features are similar to each other. The main differences are that the pressure wavefields for acoustic‐elastic coupled media show negative values even for simple geological structures unlike in acoustic media, when the Laplace damping constant is small and the water depth is shallow. The negative values may result from more complicated wave propagation in elastic media and at fluid‐solid interfaces.

Our Laplace‐domain waveform inversion algorithm is also based on the finite‐element method and logarithmic wavefields. To compute gradient direction, we apply the back‐propagation technique. Under the assumption that density is fixed, P‐ and S‐wave velocity models are inverted from the pressure data. We applied our inversion algorithm to the SEG/EAGE salt model and the numerical results showed that the Laplace‐domain waveform inversion successfully recovers the long‐wavelength structures of the P‐ and S‐wave velocity models from the noise‐free data. The models inverted by the Laplace‐domain waveform inversion were able to be successfully used as initial models in the subsequent frequency‐domain waveform inversion, which is performed to describe the short‐wavelength structures of the true models.
Application of the virtual refraction to near‐surface characterization at the Boise Hydrogeophysical Research Site‡
Authors: Nichols Josh , Mikesell Dylan , Van Wijk Kasper
Published: May 2010

Seismic interferometry is a relatively new technique to estimate the Green's function between receivers. Spurious energy, not part of the true Green's function, is produced because assumptions are commonly violated when applying seismic interferometry to field data. Instead of attempting to suppress all spurious energy, we show how spurious energy associated with refractions contains information about the subsurface in field data collected at the Boise Hydrogeophysical Research Site. By forming a virtual shot record we suppress uncorrelated noise and produce a virtual refraction that intercepts zero offset at zero time. These two features make the virtual refraction easy to pick, providing an estimate of refractor velocity. To obtain the physical parameters of the layer above the refractor we analyse the cross‐correlation of wavefields recorded at two receivers for all sources. A stationary‐phase point associated with the correlation between the reflected wave and refracted wave from the interface identifies the critical offset. By combining information from the virtual shot record, the correlation gather and the real shot record we determine the seismic velocities of the unsaturated and saturated sands, as well as the variable relative depth to the water‐table. Finally, we discuss how this method can be extended to more complex geologic models.
A strategy for automated analysis of passive microseismic data to image seismic anisotropy and fracture characteristics
Authors: Wuestefeld Andreas , Al‐Harrasi Othman , Verdon James P. , Wookey James , Kendall J. Michael
Published: May 2010

Monitoring of induced seismicity is gaining importance in a broad range of industrial operations from hydrocarbon reservoirs to mining to geothermal fields. Such passive seismic monitoring mainly aims at identifying fractures, which is of special interest for safety and productivity reasons. By analysing shear‐wave splitting it is possible to determine the anisotropy of the rock, which may be caused by sedimentary layering and/or aligned fractures, which in turn offers insight into the state of stress in the reservoir. We present a workflow strategy for automatic and effective processing of passive microseismic data sets, which are ever increasing in size. The automation provides an objective quality control of the shear‐wave splitting measurements and is based on characteristic differences between the two independent eigenvalue and cross‐correlation splitting techniques. These differences are summarized in a quality index for each measurement, allowing identification of an appropriate quality threshold. Measurements above this threshold are considered to be of good quality and are used in further interpretation. We suggest an automated inversion scheme using rock physics theory to test for best correlation of the data with various combinations of fracture density, its strike and the background anisotropy. This fully automatic workflow is then tested on a synthetic and a real microseismic data set.
Effect of wilting, silage additive, PEG treatment and tannin content on the distribution of N between different fractions after ensiling of three different sainfoin (Onobrychis viciifolia) varieties
Authors: Lorenz M. M. , Eriksson T. , Udén P.
Published: May 2010

Sainfoin (Onobrychis viciifolia) is a tanniniferous, leguminous plant that has potentially beneficial effects on protein utilization in ruminants. As ensiling causes protein breakdown and elevated levels of buffer soluble N (BSN), we studied the distribution of N before and after ensiling sainfoin. Three varieties of sainfoin were either direct‐cut and frozen directly or wilted and frozen before later ensiling in mini‐silos with and without acidification with Promyr (PM; an acidifying commercial mixture of propionic and formic acid) and with or without polyethylene glycol (PEG). Extractable tannins (ET) and protein‐bound tannins (PBT) were measured with an HCl/butanol method in an attempt to correlate tannin levels to N fractions. The sainfoin silages showed good ensiling characteristics and had relatively high concentrations of undegraded protein. The effect of wilting on BSN levels (g/kg N) was dependent on sainfoin variety (P < 0·001). PEG increased and PM decreased the level of BSN in the silages (P < 0·001). PM treatment also produced less non‐protein N and ammonia‐N (P < 0·05) as compared with no additive. Addition of PEG to the silage increased the BSN‐proportion 1·8‐ and 2·6‐fold for both DM stages. A strong tannin‐protein binding effect is, therefore, confirmed in sainfoin. However, correlations between tannin levels (ET and PBT) and BSN were poor in the (non‐PEG) silages, indicating either that the HCl/butanol method is unsuitable for measuring tannin in silages or that qualitative attributes of tannins are more relevant than quantitative. The HCl/butanol method seems therefore not to be useful to predict degradation of protein in sainfoin silages.
Re‐evaluation of the liming‐fertilization interaction in grasslands on poor and acid soils
Authors: Poozesh V. , Castillon P. , Cruz P. , Bertoni G.
Published: May 2010

The effect of surface liming on herbage production in permanent grasslands is rather uncertain. To better understand the effect of liming on the grassland, a study was made with a field experiment and a pot experiment with soil from the same field. In the field, the effects of liming and NPK fertilization on the production and composition of the vegetation were studied. In pots, the effects of liming and phosphate fertilization on different grass species were analysed. The effect of NPK fertilization (+3·96 t ha−1) on the production of the original grassland was greater than that of liming (+0·68 t ha−1), which was only observed (P < 0·05) on the unfertilized plots. Liming increased the total number of species and the proportion of dicotyledons. After replacing the semi‐natural community with Dactylis glomerata L., the effects of liming (+2·37 t ha−1) and fertilization (+6·52 t ha−1) were increased. These results, together with those of the pot trial, show the important role of phosphorus in the fertilization effect, and are interpreted as a protective effect of P against aluminium toxicity.
Interleukin‐7 promotes the survival of human CD4+ effector/memory T cells by up‐regulating Bcl‐2 proteins and activating the JAK/STAT signalling pathway
Authors: Chetoui Nizar , Boisvert Marc , Gendron Steve , Aoudjit Fawzi
Published: May 2010

Interleukin‐7 (IL‐7) is a crucial cytokine involved in T‐cell survival and development but its signalling in human T cells, particularly in effector/memory T cells, is poorly documented. In this study, we found that IL‐7 protects human CD4+ effector/memory T cells from apoptosis induced upon the absence of stimulation and cytokines. We show that IL‐7 up‐regulates not only Bcl‐2 but also Bcl‐xL and Mcl‐1 as well. Interleukin‐7‐induced activation of the janus kinase/signal transducer and activator of transcription (JAK/STAT) signalling pathway is sufficient for cell survival and up‐regulation of Bcl‐2 proteins. In contrast to previous studies with naive T cells, we found that IL‐7 is a weak activator of the phosphatidylinositol 3 kinase (PI3K)/AKT (also referred as protein kinase B) pathway and IL‐7‐mediated cell survival occurs independently from the PI3K/AKT pathway as well as from activation of the mitogen‐activated protein kinase/extracellular signal‐regulated kinase pathway. Considering the contribution of both IL‐7 and CD4+ effector/memory T cells to the pathogenesis of autoimmune diseases such as rheumatoid arthritis and colitis, our study suggests that IL‐7 can contribute to these diseases by promoting cell survival. A further understanding of the mechanisms of IL‐7 signalling in effector/memory T cells associated with autoimmune inflammatory diseases may lead to potential new therapeutic avenues.
Long‐term sealing ability of resin‐based root canal fillings
Authors: Santos J. N. , Tjäderhane L. , Ferraz C. C. , Zaia A. A. , Alves M. C. , De Goes M. F. , Carrilho M. R.
Published: May 2010

Santos J, Tjäderhane L, Ferraz C, Zaia A, Alves M, De Goes M, Carrilho M. Long‐term sealing ability of resin‐based root canal fillings. International Endodontic Journal, 43, 455–460, 2010.

Aim  To evaluate the ability of two resin‐based filling materials to provide immediate and long‐term sealing of the root canal.

Methodology  A total of eighty‐two human roots were instrumented and filled with AH Plus/gutta‐percha or Epiphany/Resilon. Root filled teeth were sealed coronally either with Coltosol or Clearfil SE Bond/Filtek Z250 or were left unsealed. The quality of root canal sealing was assessed by a fluid filtration method performed at immediate and 180‐day time intervals. Mean fluid filtration rates were analyzed by three‐way repeated measures anova and Tukey post hoc test.

Results  Specimens filled with Epiphany/Resilon exhibited higher leakage than specimens filled with AH Plus/gutta‐percha (P < 0.05), regardless of the coronal sealing condition and period of evaluation. No difference was detected between coronal restorative materials (P > 0.05), whilst leakage in teeth without any coronal restoration was significantly higher (P < 0.05). After storage, a significant decrease in leakage (P < 0.05) was observed in all experimental groups.

Conclusions  AH Plus/gutta‐percha provided superior root canal sealing at both immediate and 180‐day time periods. The presence of a coronal seal reduced leakage significantly. Storage of root filled specimens did not disturb the sealing ability of the tested materials.
Effect of alendronate on alveolar bone resorption and angiogenesis in rats with experimental periapical lesions
Authors: Xiong H. , Wei L. , Hu Y. , Zhang C. , Peng B.
Published: May 2010

Xiong H, Wei L, Hu Y, Zhang C, Peng B. Effect of alendronate on alveolar bone resorption and angiogenesis in rats with experimental periapical lesions. International Endodontic Journal, 43, 485–491, 2010.

Aim  To investigate the effects of systemically administered alendronate, one of the most potent bisphosphonates (BPs), on alveolar bone resorption and angiogenesis in rats subjected to experimental periapical lesions over two time periods.

Methodology  Forty adult Sprague‐Dawley (SD) rats were divided equally into control and experimental groups, and the pulp chambers of mandibular first molars of all rats were exposed to the oral environment to induce periapical lesions. The experimental group received daily subcutaneous injections of alendronate at a dose of 0.25 mg kg−1, whereas the control group received only the saline vehicle. These injections were initiated 1 week before the periapical lesion induction and then continued daily throughout the entire experimental period. After 2 or 4 weeks following pulp exposure, the rats were killed, and the mandibles were examined histologically for periapical bone loss area, number of microvascular vessels (NMV) and tartrate‐resistant acid phosphatase (TRAP) activity.

Results  Overall, periapical bone loss area and the number of TRAP‐positive cells (osteoclasts) were significantly decreased at 2 and 4 weeks, respectively, after daily subcutaneous injection of alendronate compared with the control group (P < 0.05). There was no significant decrease change in NMV (P > 0.05).

Conclusions  Administration of alendronate to rats might inhibit alveolar bone resorption associated with periapical disease, which might not lead to impairment of angiogenesis. However, because of the differences between rats and humans, one has to consider the possible consequences of this treatment in the clinic.
Differential expression of neuropeptide Y Y1 receptors during pulpal inflammation
Authors: Rethnam S. , Raju B. , Fristad I. , Berggreen E. , Heyeraas K. J.
Published: May 2010

Rethnam S, Raju B, Fristad I, Berggreen E, Heyeraas KJ.Differential expression of neuropeptide Y Y1 receptors during pulpal inflammation.International Endodontic Journal,43, 492–498, 2010.

Aim To study the pattern of neuropeptide Y (NPY) Y1 receptor (Y1R) localization in the normal dental pulp and during different stages of pulpal inflammation. The hypothesis was that the expression of Y1R varies during different stages of pulpitis.

Methodology Pulp exposure injury was made on first molar teeth of Sprague–Dawley rats. Animals were killed 3, 7, 14, 21, 28 and 46 days after pulp exposure. Jaws were removed, decalcified and processed for immunohistochemistry for identification and localization of Y1R. Double labelling was performed using antibodies for laminin, CD43 and CD4 with Y1R. RT‐PCR was performed to verify gene expression of Y1R in dental pulp and trigeminal ganglion.

Results RT‐PCR revealed the presence of NPY‐Y1R in the dental pulp, trigeminal ganglion and gingiva. With immunohistochemistry, in control rats, Y1R was mainly located in capillaries and small vessels in the dental pulp as well as in the odontoblastic layer. No YIR was noted on immune cells in normal pulp. In pulpitis, immune cells such as CD43+ granulocytes and CD4+ lymphocytes expressed Y1R. As pulpitis progressed, there was a significant decrease in number of blood vessels expressing Y1R in the odontoblast layer when compared to control rat pulp.

Conclusions This study gives evidence that Y1R is a modulator of pulpal inflammation.
Radiographic quality of root fillings performed by undergraduate students
Authors: Khabbaz M. G. , Protogerou E. , Douka E.
Published: May 2010

Khabbaz MG, Protogerou E, Douka E. Radiographic quality of root fillings performed by undergraduate students. International Endodontic Journal, 43, 499–508, 2010.

Aim  To evaluate the radiographic technical quality of root fillings and the incidence of iatrogenic errors in treatment provided by undergraduate students.

Methodology  Endodontic records and periapical radiographs of 759 root filled teeth were selected following exclusion of 25 (3.3%). A final total of 734 cases were used to assess the technical quality of root fillings in 1109 root canals performed by 4th and 5th undergraduate students. Two variables were examined: the length and the density of the fillings, categorized as acceptable and non‐acceptable. The presence of ledges, root perforation, foramen perforation and fractured instruments were also investigated. Chi‐square tests were used for statistical analysis.

Results  Acceptable root fillings were found in 55% of canals. More `acceptable' root fillings occurred in maxillary compared to mandibular teeth (58 and 51% respectively) (P < 0.05) and in anterior compared with premolar (71 and 61%) and molar root canals (39% respectively) (P < 0.05). Insignificantly more (5.4%) acceptable root fillings were provided by 5th year students. Most of the underfillings and iatrogenic errors were detected in molars for both academic years (53 and 65% respectively). Ledges were noted in 55% of cases treated by both academic years and significantly more foramen perforations were detected in the 5th compared with the 4th year students (P < 0.05).

Conclusions  There is a need to improve the technical quality of root fillings performed by the undergraduate students, especially in molar teeth.
Efficacy of different irrigants in the removal of calcium hydroxide from root canals
Authors: Rödig T. , Vogel S. , Zapf A. , Hülsmann M.
Published: May 2010

Rödig T, Vogel S, Zapf A, Hülsmann M. Efficacy of different irrigants in the removal of calcium hydroxide from root canals. International Endodontic Journal. 43, 519–527, 2010.

Aim  To compare the efficacy of different solutions (1% sodium hypochlorite, 10% citric acid and 20% EDTA) in the removal of calcium hydroxide from root canals.

Methodology  One hundred single‐rooted maxillary incisors were prepared to size 50 and split longitudinally. Two standardized grooves were cut into the apical and coronal part of the root canal dentine and filled with calcium hydroxide. The reassembled teeth were irrigated with a syringe and a size 30 needle using the following irrigants: (i) 20% EDTA, (ii) 10% citric acid, (iii) 1% NaOCl, (iv) 10% citric acid + 1% NaOCl, (v) 20% EDTA + 1% NaOCl and (vi) water (control). Volume of irrigant was 20 mL in each group, and irrigation time was 5 min. Evaluation of cleanliness of the blinded specimens was performed by two calibrated observers under a microscope with 30× magnification using a four‐grade scoring system as described by van der Sluis et al. (2007). Statistical evaluation was performed using a SAS‐macro for non‐parametric multifactorial analysis (P < 0.05).

Results  The best results were found for irrigation with EDTA and citric acid, whereas NaOCl and water showed the least effect. The combinations of irrigants did not result in improvement in terms of cleanliness.

Conclusions  None of the irrigants nor their respective combinations were able to completely remove the calcium hydroxide. Chelating agents such as citric acid and EDTA showed the best results. The combination of chelators and NaOCl did not result in significant improvement of calcium hydroxide removal.
In vitro antibacterial action of Tetraclean, MTAD and five experimental irrigation solutions
Authors: Pappen F. G. , Shen Y. , Qian W. , Leonardo M. R. , Giardino L. , Haapasalo M.
Published: May 2010

Pappen FG, Shen Y, Qian W, Leonardo MR, Giardino L, Haapasalo M.
In vitro antibacterial action of Tetraclean, MTAD and five experimental irrigation solutions. International Endodontic Journal, 43, 528–535, 2010.

Aim  To investigate the antibacterial effect of Tetraclean, MTAD and five experimental irrigants using both direct exposure test with planktonic cultures and mixed‐species in vitro biofilm model.

Methodology  Tetraclean, MTAD and five experimental solutions that were modifications of existing formulae including MTAD + 0.01% cetrimide (CTR), MTAD + 0.1% CTR, MTAC‐1 (Tween 80 replaced by 0.01% CTR in MTAD), MTAC‐2 (Tween 80 replaced by 0.1% CTR) and MTAD‐D (MTAD without the Tween 80 and no CTR added) were used as disinfectants in the experiments. In the direct exposure test, a suspension of Enterococcus faecalis was mixed with each of the solutions. After 0.5, 1, 3 and 10 min, an inactivator was added and the number of surviving bacteria was calculated. A mixed‐species biofilm from subgingival plaque bacteria was grown in brain heart infusion broth in anaerobic conditions on synthetic hydroxyapatite discs. Two‐week‐old biofilms were exposed to the solutions for 0.5, 1 and 3 min. The samples were observed by confocal laser scanning microscopy after bacterial viability staining. The scans were quantitatively analysed, and the volume of killed cells of all cells was calculated for each medicament.

Results  Tetraclean and MTAC‐2 (0.1% CTR) killed planktonic E. faecalis in <30 s. Complete killing of bacteria required 1 min by MTAC‐1, 3 min by MTAD + 0.1% CTR and 10 min by MTAD, MTAD‐D and MTAD + 0.01% CTR. In the biofilm test, there were significant differences in microbial killing between the different solutions and times of exposure (P < 0.005). MTAC‐2 showed the best performance, killing 71% of the biofilm bacteria in 3 min, followed by MTAC‐1 and Tetraclean. MTAD and the three MTAD modifications demonstrated the lowest antibacterial activity.

Conclusion  Tetraclean was more effective than MTAD against E. faecalis in planktonic culture and in mixed‐species in vitro biofilm. CTR improved the antimicrobial properties of the solutions, whereas Tween 80 seemed to have a neutral or negative impact on their antimicrobial effectiveness.
Repeated vicariance of Eurasian songbird lineages since the Late Miocene
Authors: Voelker Gary
Published: May 2010

Aim  The evolution of avian speciation patterns across much of Eurasia is under‐explored. Excepting phylogeographic patterns of single species, or speciation involving the Himalayas, there has been no attempt to understand the evolution of avian distributional patterns across the rest of the continent. Within many genera there is a pattern of (presumed) sister species occurring in adjacent areas (western, eastern or southern Eurasia), yet this pattern cannot be explained by existing biogeographic barriers. My aim was to examine the possible role of climate‐driven vicariance events in generating avian distributions.

Location  Eurasia.

Methods  I constructed a molecular phylogeny of Phoenicurus redstarts, and assembled phylogenetic data from published studies of seven other Eurasian bird genera. On each phylogeny, I assessed the distributional patterns of species and clades relative to refugial areas in western, eastern and southern Eurasia. I also estimated the timing of lineage divergences via a molecular clock, to determine whether distributional patterns can be explained by well‐defined periods of climate change in Eurasia that are recorded from dated sediments in the Chinese Loess Plateau.

Results  Species relationships in a well‐supported phylogeny of Phoenicurus show a pattern of distributions consistent with repeated speciation in major refugial areas, where one lineage is isolated in a single area of Eurasia relative to its sister lineage. This same pattern is evident in Eurasian Turdus thrushes, and six additional avian genera distributed across Eurasia. Molecular clock dating indicates that divergences within each genus are the result of multiple rounds of speciation in refugia through time, during major climate‐driven episodes of vicariance.

Main conclusions  Analyses revealed substantial evidence supporting a repeated, non‐random pattern of speciation within and across eight songbird lineages since the Late Miocene. The pattern of speciation supports a model of isolation in refugia during major episodes of vicariance, specifically periods of either intensified desertification of Central Asia or Eurasian glacial cycles. The densely sampled clades used here preclude inter‐continental dispersal as an alternative explanation for distributions. The signature of climate‐driven vicariance across epochs is, given the absence of extant biogeographic barriers, a suitable hypothesis to explain major lineage divergences in widely distributed Eurasian songbird lineages.
Using malpractice claims to identify risk factors for neurological impairment among infants following non‐reassuring fetal heart rate patterns during labour
Authors: Kesselheim Aaron S. , November Martin T. , Lifford Karen L. , McElrath Thomas F. , Puopolo Ann L. , Orav E. John , Studdert David M.
Published: May 2010

Rationale, Aims and Objectives  We sought to use a novel case‐selection methodology to identify antenatal or intrapartum risk factors associated with neonatal neurological impairment following non‐reassuring fetal heart rate patterns during labour.

Method  We used a retrospective case–control design with bivariate and multivariate conditional logistic regression. Cases were births in which electronic fetal monitoring (EFM) showed non‐reassuring patterns and the infant had neurological disability. Controls were births in which EFM was non‐reassuring but the infant was born healthy. We identified 36 cases from among malpractice claims filed with a liability insurer in Massachusetts between 1985 and 2001 and randomly selected 70 controls, matching them to cases by hospital, birth date and gestational age.

Results  More cases had maternal antenatal vaginal bleeding (P = 0.004), a prolonged latent phase or protracted dilation during the first stage of labour (P = 0.03), and protracted descent or prolonged second stage (P = 0.01). More cases also had minimal variability on EFM on admission (P = 0.02) and during the second stage (P = 0.02). Multivariate analysis highlighted three significant predictors of neurological injury following complicated labour: antenatal vaginal bleeding (OR = 27.1), prolonged latent phase or protracted dilation in the first stage (OR = 4.0) and EFM showing minimal variability in the first stage (OR = 4.3).

Conclusion  These promising initial findings suggest that future research into outcomes from complicated labour with non‐reassuring heart rate patterns should focus on maternal history of vaginal bleeding, slow labour and minimal variability on EFM.
Identifying and prioritizing uncertainties: patient and clinician engagement in the identification of research questions
Authors: Elwyn Glyn , Crowe Sally , Fenton Mark , Firkins Lester , Versnel Jenny , Walker Samantha , Cook Ivor , Holgate Stephen , Higgins Bernard , Gelder Colin
Published: May 2010

Background  To arrive at an agreed, prioritized ranking of treatment uncertainties in asthma that need further research, by developing a collaboration of patients, carers and clinicians, facilitated by the James Lind Alliance Working Partnership between Asthma UK and the British Thoracic Society.

Methods  A four‐step procedure: (1) establish a collaborative Working Partnership; (2) identify and collect treatment uncertainties by using a patient survey and analysing existing systematic reviews, clinical guidelines and query‐answering services; (3) categorize uncertainties; and (4) convene a workshop using a nominal group process to establish a ranked prioritization of treatment uncertainties in asthma.

Findings  Agreement and rankings were reached for 10 treatment uncertainties. The highest was given to the uncertainty surrounding the adverse effects of inhaled and oral steroids. The top three priorities dealt with clinical management issues, where uncertainties still exist, namely concerns about the side effects of inhaled and oral steroids, how to manage asthma when other illnesses exist or how to rely on personal decisions in an ever‐changing illness (self‐management).

Interpretation  The key outcome is the generation of a prioritized list of treatment uncertainties in asthma, agreed by a collaboration of patients and health professionals, to inform the commissioning of new research. Such a large number of patient‐identified treatment uncertainties had not previously been identified in the literature, an indication perhaps that asthma self‐management is a neglected research area. Whether the results have an influence of research funding decisions is not yet known.
Scintillation SE detector for variable pressure scanning electron microscopes
Authors: JIRÁK J. , NEDĚLA V. , ČERNOCH P. , ČUDEK P. , RUNŠTUK J.
Published: May 2010

We present results obtained with a new scintillation detector of secondary electrons for the variable pressure scanning electron microscope. A detector design is based on the positioning of a single crystal scintillator within a scintillator chamber separated from the specimen chamber by two apertures. This solution enables us to decrease the pressure to several Pa in the scintillator chamber while the pressure in the specimen chamber reaches values of about 1000 Pa (7.5 Torr). Due to decreased pressure, we can apply a potential of the order of several kV to the scintillator, which is necessary for the detection of secondary electrons. Simultaneously, the two apertures at appropriate potentials of the order of several hundreds of volts create an electrostatic lens that allows electrons to pass from the specimen chamber to the scintillator chamber. Results indicate a promising utilization of this detector for a wide range of specimen observations.
Rapid FlAsH labelling in the budding yeast Saccharomyces cerevisiae
Authors: WURM C. A. , SUPPANZ I. E. , STOLDT S. , JAKOBS S.
Published: May 2010

Live cell imaging of protein distributions is an essential tool in modern cell biology. It relies on the functional labelling of a host protein with a fluorophore, which may either be a genetically fused fluorescent protein or an organic dye binding to the host protein. The biarsenical‐tetracysteine system or ‘FlAsH‐labelling’, is based on the high affinity interaction between a biarsenical probe and a small protein tag. This approach has been successfully used for live cell imaging in the budding yeast Saccharomyces cerevisiae. However, the established labelling protocols require a lengthy overnight incubation of the cells with the dye under tightly controlled growth conditions, which severely limits the use of this approach. In this study, we characterize an efficient method for introducing FlAsH‐EDT2 into live budding yeast cells using standard electroporation. The labelling time is reduced from more than 12 h to less than 1 h without compromising the labelling efficiency or cell viability. This approach may be used for cells in different growth phases or grown under different conditions. It may be further extended to other small high affinity probes, thus opening up new possibilities for labelling in budding yeast.
Evaluating sharpness functions for automated scanning electron microscopy
Authors: RUDNAYA M.E. , MATTHEIJ R.M.M. , MAUBACH J.M.L.
Published: May 2010

Fast and reliable autofocus techniques are an important topic for automated scanning electron microscopy. In this paper, different autofocus techniques are discussed and applied to a variety of experimental through‐focus series of scanning electron microscopy images with different geometries. The procedure of quality evaluation is described, and for a variety of scanning electron microscope samples it is demonstrated that techniques based on image derivatives and Fourier transforms are in general better than statistical, intensity and histogram‐based techniques. Further, it is shown that varying of an extra parameter can dramatically increase quality of an autofocus technique.
An alternative route to nitric oxide resistance
Authors: Spiro Stephen
Published: May 2010

Vibrio fischeri is a bioluminescent bacterium that enters into a symbiosis with the bobtail squid Euprymna scolopes. The bacterium colonizes a specialized light organ, in which it generates light that might help the squid to hide its silhouette from animals beneath it. Previous studies have shown that the host nitric oxide (NO) synthase is active during colonization, suggesting that V. fischeri symbionts are exposed to NO. Thus, NO might play a role in regulating the symbiosis, a role possibly analogous to that of NO in the interaction between some pathogens and their hosts. One possibility is that NO helps to exclude other species from the light organ, in which case, the response of V. fischeri to NO is of considerable interest. In this issue of Molecular Microbiology, Dunn et al. report that V. fischeri produces an NO‐inducible and NO‐resistant alternative oxidase (Aox) that allows respiration to continue in the presence of NO concentrations that are inhibitory to the conventional respiratory oxidases. This is an important step towards a better understanding of the role that NO plays in the Vibrio‐squid symbiosis, and provides the first indication of a physiological function for a bacterial homologue of the plant Aox.
Cutaneous progressive angiomatosis on the muzzle of a dog, treated by laser photocoagulation therapy
Authors: Olivieri Lara , Nardini Giordano , Pengo Graziano , Abramo Francesca
Published: May 2010

A 10‐year old, female, cocker spaniel was presented with a bleeding lesion on the muzzle and nasal planum observed initially as a small raised nodule at 1 year of age. This became clinically more pronounced at 5 years of age, progressively enlarged thereafter and was diagnosed initially as a vascular tumour. On clinical examination, multiple confluent nodules of variable diameter (0.5–1.5 cm) deformed the outline of the nose. Histological examination revealed a progressive angiomatosis with multifocal infiltrative growth of increased dermal vascularization with different sized and type of anastomosing vessels lined by a monolayer of reactive endothelium, often arranged as papillary projections into the lumina. Partial nosectomy, the first therapeutic approach, was unsuccessful and the lesion relapsed within 6 months. However, subsequent laser photocoagulation therapy provided a good cosmetic outcome, no relapse 1 year later, and appeared to provide an effective alternative to ablation surgery.
Heterogeneity ofStaphylococcus pseudintermediusisolates from atopic and healthy dogs
Authors: Fazakerley Jennifer , Williams Nicola , Carter Stuart , McEwan Neil , Nuttall Tim
Published: May 2010

Staphylococcus pseudintermediusis part of the normal canine flora but frequently causes pyoderma in canine atopic dermatitis (AD). This study aimed to determine whether particularS. pseudintermediusstrains were associated with AD and/or pyoderma. Ninety‐sixS. pseudintermediusisolates from the ear, nares, perineum and lesions of 21 atopic and 16 healthy dogs were lysed with proteinase K and digested with 40 USmaI. Restriction products were separated using pulsed‐field gel electrophoresis (PFGE) with an OxfordS. aureuscontrol and lambda‐ladder DNA concatomer markers. A dendrogram was constructed by the unweighted pair group method. All isolates showed a ≥56% similarity coefficient. Nine distinct PFGE clusters were identified, as follows: five from both atopic and healthy dogs; three from atopic dogs only; and one from healthy dogs only. Nine clusters were isolated from the nares, eight from the perineum, five from the ears and six from pyoderma lesions. There were no significant differences in the frequency of isolation from atopic or healthy skin, body sites or infected lesions for any of the clusters. Two of six healthy dogs and 18 of 20 atopic dogs with multiple isolates had closely related isolates (less than three band differences) at more than one sampling site. Isolates from pyoderma lesions were closely related to at least one mucosal isolate in 11 of 16 dogs.Staphylococcus pseudintermediusisolates appear to be heterogeneous, and colonization or infection of atopic skin was not associated with any particular strain or cluster of strains.
Variability of estradiol concentration in normal dogs
Authors: Frank Linda A. , Mullins Rebekah , Rohrbach Barton W.
Published: May 2010

Estradiol concentrations are evaluated in canine serum as part of an adrenal panel used to diagnose atypical Cushing’s syndrome and other endocrine abnormalities. Estradiol concentrations are often elevated in dogs without clinical signs of hyperestrogenism, and the significance of this elevation is unknown. The purpose of this study was to estimate the variation in estradiol concentrations in normal dogs. Ten neutered male and female dogs were enrolled in the study. Blood was collected from each dog at 2 h intervals, four times during a given day. This was repeated approximately 1 (week 2) and 5 weeks later (week 6). There was no attempt for a given dog to be started at the exact time or day each week. Results showed that estradiol concentrations ranged from 44.6 to 120.3 pg/mL with a mean of 70.4 pg/mL, which is greater than the upper limit of normal for our laboratory (69 pg/mL). The mean difference between the highest and lowest concentrations for each dog was 28.8 pg/mL, with a range of 12.5–53.5 pg/mL. Mean estradiol concentrations from week 6 (63.2 pg/mL) were significantly lower than those from week 1 (71.4 pg/mL;P = 0.015) and week 2 (76.5 pg/mL;P = 0.0004). These data show a wide range of variability in estradiol concentration both within and between dogs and that these measurements often exceed the normal ranges established by the laboratory. Therefore, diagnosis of hyperestrogenism or atypical Cushing’s syndrome based on increased estradiol concentrations should require compatible clinical presentation of hyperestrogenism together with elevated serum estradiol.
The role of plastidial glucose‐6‐phosphate/phosphate translocators in vegetative tissues of Arabidopsis thaliana mutants impaired in starch biosynthesis
Authors: Kunz H. H. , Häusler R. E. , Fettke J. , Herbst K. , Niewiadomski P. , Gierth M. , Bell K. , Steup M. , Flügge U.‐I. , Schneider A.
Published: May 2010

Arabidopsis thaliana mutants impaired in starch biosynthesis due to defects in either ADP glucose pyrophosphorylase (adg1‐1), plastidic phosphoglucose mutase (pgm) or a new allele of plastidic phosphoglucose isomerase (pgi1‐2) exhibit substantial activity of glucose‐6‐phosphate (Glc6P) transport in leaves that is mediated by a Glc6P/phosphate translocator (GPT) of the inner plastid envelope membrane. In contrast to the wild type, GPT2, one of two functional GPT genes of A. thaliana, is strongly induced in these mutants during the light period. The proposed function of the GPT in plastids of non‐green tissues is the provision of Glc6P for starch biosynthesis and/or the oxidative pentose phosphate pathway. The function of GPT in photosynthetic tissues, however, remains obscure. The adg1‐1 and pgi1‐2 mutants were crossed with the gpt2‐1 mutant defective in GPT2. Whereas adg1‐1/gpt2‐1 was starch‐free, residual starch could be detected in pgi1‐2/gpt2‐1 and was confined to stomatal guard cells, bundle sheath cells and root tips, which parallels the reported spatial expression profile of AtGPT1. Glucose content in the cytosolic heteroglycan increased substantially in adg1‐1 but decreased in pgi1‐2, suggesting that the plastidic Glc6P pool contributes to its biosynthesis. The abundance of GPT2 mRNA correlates with increased levels of soluble sugars, in particular of glucose in leaves, suggesting induction by the sugar‐sensing pathway. The possible function of GPT2 in starch‐free mutants is discussed in the background of carbon requirement in leaves during the light–dark cycle.
Two differentially expressed MATE factor genes from apple complement the Arabidopsis transparent testa12 mutant
Authors: Frank S. , Keck M. , Sagasser M. , Niehaus K. , Weisshaar B. , Stracke R.
Published: May 2010

Proanthocyanidins (PAs) are a class of flavonoids with numerous functions in plant ecology and development, including protection against microbial infection, animal foraging and damage by UV light. PAs are also beneficial in the human diet and livestock farming, preventing diseases of the cardiovascular system and lowering the risk of cancer, asthma and diabetes. Apples (Malus x domestica Borkh.) are naturally rich in flavonoids, but the flavonoid content and composition varies significantly between cultivars. In this work, we applied knowledge from the model plant Arabidopsis thaliana, for which the main features of flavonoid biosynthesis have been elucidated, to investigate PA accumulation in apple. We identified functional homologues of the Multidrug And Toxic compound Extrusion (MATE) gene TRANSPARENT TESTA12 from A. thaliana using a comparative genomics approach. MdMATE1 and MdMATE2 were differentially expressed, and the function of the encoded proteins was verified by complementation of the respective A. thaliana mutant. In addition, MdMATE genes have a different gene structure in comparison to homologues from other species. Based on our findings, we propose that MdMATE1 and MdMATE2 are vacuolar flavonoid/H+‐antiporters, active in PA accumulating cells of apple fruit. The identification of these flavonoid transporter genes expands our understanding of secondary metabolite biosynthesis and transport in apple, and is a prerequisite to improve the nutritional value of apples and apple‐derived beverages.