bluuebunny/crossref_metadata_2025 huggingface dataset:
Rationale and design of the Eplerenone in Mild Patients Hospitalization And SurvIval Study in Heart Failure (EMPHASIS‐HF)
Authors: Zannad Faiez , McMurray John J.V. , Drexler Helmut , Krum Henry , van Veldhuisen Dirk J. , Swedberg Karl , Shi Harry , Vincent John , Pitt Bertram
Published: April 2010

Aims

In chronic heart failure (HF), aldosterone antagonists have been shown to improve survival in patients with low ejection fraction and moderate‐to‐severe symptoms [New York Heart Association (NYHA) classes III and IV]. Efficacy of these agents was also shown when they were administered to patients with left ventricular dysfunction and signs and symptoms of CHF early after acute myocardial infarction. It is not known whether the selective aldosterone antagonist eplerenone can improve outcomes in mildly symptomatic patients. The Eplerenone in Mild Patients Hospitalization And SurvIval Study in Heart Failure (EMPHASIS‐HF) was designed to evaluate the effect of eplerenone on mortality and morbidity in patients with chronic systolic HF in NYHA class II.

Methods

Approximately 3100 patients with ejection fraction ≤30% and estimated glomerular filtration rate ≥30 mL/min/1.73 m2 will be recruited. Patients are randomized 1:1 to double‐blind eplerenone or placebo in addition to standard chronic HF therapy. Doses are adjusted from 25 mg every other day to 50 mg daily, depending on serum potassium. The primary endpoint is a composite of time to cardiovascular death or first hospital admission for worsening HF, whichever occurs first.

Conclusion

The study will be complete when approximately 813 subjects experience a primary endpoint.

Clinical Trials.gov. NCT00232180.
Acute effects of urocortin 2 on cardiac function and propensity for arrhythmias in an animal model of hypertension‐induced left ventricular hypertrophy and heart failure
Authors: Meili‐Butz Silvia , Bühler Katrin , John Dietlinde , Buser Peter , Vale Wylie W. , Peterson Kirk L. , Brink Marijke , Dieterle Thomas
Published: April 2010

Aims

To test acute effects of the corticotropin‐releasing factor‐related peptide urocortin 2 (Ucn2) on left ventricular (LV) function and the propensity for ventricular arrhythmias in the isolated heart of an animal model of hypertension‐induced heart failure.

Methods and results

Hearts from Dahl salt‐sensitive rats with severe LV dysfunction were perfused according to Langendorff. Left ventricular developed pressure and the positive and negative derivatives of LV pressure were analysed before and after perfusion with Ucn2 (n = 15) or normal perfusion solution (control, n = 9). Intracellular calcium cycling parameters were assessed by surface fluorometry. Furthermore, monophasic action potential duration (MAPD) and ventricular fibrillation threshold (VFT) were determined, the latter by a train‐of‐pulses method at increasing voltage to scan the vulnerable period of repolarization. Urocortin 2 significantly affected intracellular calcium cycling and improved LV contractile function and relaxation. Compared with baseline values, Ucn2 significantly decreased MAPD at 30, 50, and 90% repolarization and significantly increased VFT compared with baseline values. No changes were observed in control experiments.

Conclusion

Administration of Ucn2 rapidly improves LV function and increases VF threshold in failing, isolated rat hearts with increased propensity for ventricular arrhythmias. These observations suggest a potential use of Ucn2 as a safe and novel agent for the treatment of acute heart failure.
Effect of General Anesthesia and Orthopedic Surgery on Serum Tryptase
Authors: Garvey Lene H. , Bech Birgitte , Mosbech Holger , Krøigaard Mogens , Belhage Bo , Husum Bent , Poulsen Lars K.
Published: April 2010

Background

Mast cell tryptase is used clinically in the evaluation of anaphylaxis during anesthesia, because symptoms and signs of anaphylaxis are often masked by the effect of anesthesia. No larger studies have examined whether surgery and anesthesia affect serum tryptase. The aim of this study was to investigate the effect of anesthesia and surgery on serum tryptase in the absence of anaphylaxis.

Methods

The study included 120 patients (median age, 54 yr; range, 19-94 yr) undergoing elective orthopedic surgery in general anesthesia. Exclusion criteria were allergic reactions during this or previous anesthesia, hematologic disease, or high-dose corticosteroid treatment. Blood samples for tryptase analysis (ImmunoCAP; Phadia, Uppsala, Sweden) were drawn shortly before anesthesia and after anesthesia and surgery.

Results

Median duration of anesthesia was 105 min (range, 44-263 min). Median interval between blood samples was 139 min (range, 39-370 min). Mean tryptase before surgery was 5.01 microg/l, with a mean decrease of 0.55 microg/l (P < 0.0001; 95% CI, 0.3-0.8) postoperatively. All patients received intravenous fluid (median value 750 ml; range, 200-2000 ml) perioperatively. There was no significant effect of gender, age, American Society of Anesthesiologist's physical status classification, or self-reported allergy on serum tryptase.

Conclusions

Serum tryptase shows small intraindividual variation in the absence of anaphylaxis. A small decrease was observed postoperatively, likely due to dilution by intravenous fluid. On suspected anaphylaxis during anesthesia, tryptase values, even within the normal reference interval, should, when possible, be compared with the patient's own basal level taken more than 24 h after the reaction.
Cytosol-Localized Heat Shock Factor-Binding Protein, AtHSBP, Functions as a Negative Regulator of Heat Shock Response by Translocation to the Nucleus and Is Required for Seed Development in Arabidopsis
Authors: Hsu Shih-Feng , Lai Hui-Chuan , Jinn Tsung-Luo
Published: April 2010

Heat shock response (HSR) is a universal mechanism in all organisms. It is under tight regulation by heat shock factors (HSFs) and heat shock proteins (HSPs) after heat shock (HS) to prevent stress damage. On the attenuation of HSR, HSP70 and HSF Binding Protein1 (HSBP1) interact with HSF1 and thus dissociate trimeric HSF1 into an inert monomeric form in humans. However, little is known about the effect of HSBP with thermal stress in plants. This report describes our investigation of the role of AtHSBP in Arabidopsis (Arabidopsis thaliana) by genetic and molecular approaches. AtHSBP was heat inducible and ubiquitously expressed in all tissues; AtHSBP was also crucial for seed development, as demonstrated by AtHSBP-knockout lines showing seed abortion. Thermotolerance results showed that AtHSBP participates in acquired thermotolerance but not basal thermotolerance and is a negative regulator of HSR. Subcellular localization revealed that the cytosol-localized AtHSBP translocated to the nucleus in response to HS. Protoplast two-hybrid assay results confirmed that AtHSBP interacts with itself and with the HSFs, AtHSFA1a, AtHSFA1b, and AtHSFA2. AtHSBP also negatively affected AtHSFA1b DNA-binding capacity in vitro. Quantitative polymerase chain reaction and western-blot analysis demonstrated that altered levels of AtHSBP lead to differential HSP expression, mainly during the recovery from HS. These studies provide a new insight into HSBP in plants and reveal that AtHSBP is a negative regulator of HSR and required for seed development.
VND-INTERACTING2, a NAC Domain Transcription Factor, Negatively Regulates Xylem Vessel Formation in Arabidopsis
Authors: Yamaguchi Masatoshi , Ohtani Misato , Mitsuda Nobutaka , Kubo Minoru , Ohme-Takagi Masaru , Fukuda Hiroo , Demura Taku
Published: April 2010

The Arabidopsis thaliana NAC domain transcription factor VASCULAR-RELATED NAC-DOMAIN7 (VND7) acts as a master regulator of xylem vessel differentiation. To understand the mechanism by which VND7 regulates xylem vessel differentiation, we used a yeast two-hybrid system to screen for proteins that interact with VND7 and identified cDNAs encoding two NAC domain proteins, VND-INTERACTING1 (VNI1) and VNI2. Binding assays demonstrated that VNI2 effectively interacts with VND7 and the VND family proteins, VND1-5, as well as with other NAC domain proteins at lower affinity. VNI2 is expressed in both xylem and phloem cells in roots and inflorescence stems. The expression of VNI2 overlaps with that of VND7 in elongating vessel precursors in roots. VNI2 contains a predicted PEST motif and a C-terminally truncated VNI2 protein, which lacks part of the PEST motif, is more stable than full-length VNI2. Transient reporter assays showed that VNI2 is a transcriptional repressor and can repress the expression of vessel-specific genes regulated by VND7. Expression of C-terminally truncated VNI2 under the control of the VND7 promoter inhibited the normal development of xylem vessels in roots and aerial organs. These data suggest that VNI2 regulates xylem cell specification as a transcriptional repressor that interacts with VND proteins and possibly also with other NAC domain proteins.
Arrangement of Photosystem II and ATP Synthase in Chloroplast Membranes of Spinach and Pea
Authors: Daum Bertram , Nicastro Daniela , Austin Jotham , McIntosh J. Richard , Kühlbrandt Werner
Published: April 2010

We used cryoelectron tomography to reveal the arrangements of photosystem II (PSII) and ATP synthase in vitreous sections of intact chloroplasts and plunge-frozen suspensions of isolated thylakoid membranes. We found that stroma and grana thylakoids are connected at the grana margins by staggered lamellar membrane protrusions. The stacking repeat of grana membranes in frozen-hydrated chloroplasts is 15.7 nm, with a 4.5-nm lumenal space and a 3.2-nm distance between the flat stromal surfaces. The chloroplast ATP synthase is confined to minimally curved regions at the grana end membranes and stroma lamellae, where it covers 20% of the surface area. In total, 85% of the ATP synthases are monomers and the remainder form random assemblies of two or more copies. Supercomplexes of PSII and light-harvesting complex II (LHCII) occasionally form ordered arrays in appressed grana thylakoids, whereas this order is lost in destacked membranes. In the ordered arrays, each membrane on either side of the stromal gap contains a two-dimensional crystal of supercomplexes, with the two lattices arranged such that PSII cores, LHCII trimers, and minor LHCs each face a complex of the same kind in the opposite membrane. Grana formation is likely to result from electrostatic interactions between these complexes across the stromal gap.
Scatter: software for the analysis of nano- and mesoscale small-angle scattering
Authors: Förster S. , Apostol L. , Bras W.
Published: April 2010

Scatteris a new software for analysis, modeling and fitting of one- and two-dimensional small-angle scattering data of non-ordered, partially ordered or fully ordered nano- and mesoscale structures. The calculations are based on closed analytical expressions for the scattering intensity, enabling efficient evaluation of form factors and structure factors. The software allows one to sequentially fit large series of scattering curves and scattering patterns automatically. It provides further tools for data loading, beam centering, calibration, zooming, binning, lattice identification, calculation of density profiles and size distributions, and visualization of real-space structures. Presentations of experimental and calculated data can be saved as is for presentations or exported for further graphical or mathematical treatment.
Implementation of statistical dynamic diffraction theory for defective semiconductor heterostructure modelling
Authors: Shreeman P. K. , Matyi R. J.
Published: April 2010

Statistical dynamic diffraction theory (SDDT) provides the ability to model defect-induced structures in high-resolution X-ray diffraction analyses by incorporating both coherent (dynamic) and incoherent (kinematic) scattering. Current treatments of SDDT are mathematically intensive and may not provide sufficient detail regarding the implementation of the theory in practice. This paper discusses the implementation of SDDT and the modifications that allow for successful SDDT analyses of fully relaxed SiGe on Si.
Coding behavioural data for cladistic analysis: using dynamic homology without parsimony
Authors: Japyassú Hilton F. , Machado Fabio de A.
Published: April 2010

Many of the controversies around the concept of homology rest on the subjectivity inherent to primary homology propositions. Dynamic homology partially solves this problem, but there has been up to now scant application of it outside of the molecular domain. This is probably because morphological and behavioural characters are rich in properties, connections and qualities, so that there is less space for conflicting character delimitations. Here we present a new method for the direct optimization of behavioural data, a method that relies on the richness of this database to delimit the characters, and on dynamic procedures to establish character state identity. We use between‐species congruence in the data matrix and topological stability to choose the best cladogram. We test the methodology using sequences of predatory behaviour in a group of spiders that evolved the highly modified predatory technique of spitting glue onto prey. The cladogram recovered is fully compatible with previous analyses in the literature, and thus the method seems consistent. Besides the advantage of enhanced objectivity in character proposition, the new procedure allows the use of complex, context‐dependent behavioural characters in an evolutionary framework, an important step towards the practical integration of the evolutionary and ecological perspectives on diversity.

© The Willi Hennig Society 2010.
Assessing genetic diversity of Vietnamese local chicken breeds using microsatellites
Authors: Cuc N. T. K. , Simianer H. , Eding H. , Tieu H. V. , Cuong V. C. , Wollny C. B. A. , Groeneveld L. F. , Weigend S.
Published: April 2010

This study aimed to assess genetic diversity within and between nine Vietnamese local chicken breeds and two Chinese breeds included for comparison. Genotyping 29 microsatellites revealed high diversity of both Vietnamese and Chinese breeds. Cluster analysis using the structure software suggested six clusters as the most likely grouping of the 11 breeds studied. These groups encompassed four homogeneous clusters, one formed by the two Chinese breeds and the other three representing a single breed each: the Mekong Delta breed Ac, the South Central Coast breed Choi, and the Red River Delta breed Dong Tao. The six remaining breeds formed two additional admixed clusters.
Effects of p.C430S polymorphism in the PPARGC1A gene on muscle fibre type composition and meat quality in Yorkshire pigs
Authors: Kim J. M. , Lee K. T. , Lim K. S. , Park E. W. , Lee Y. S. , Hong K. C.
Published: April 2010

The peroxisome proliferator‐activated receptor‐gamma coactivator‐1A (encoded by PPARGC1A) is involved in the formation of type I fibres. Therefore, the PPARGC1A gene can be considered as a functional candidate gene for muscle fibre type composition and meat quality in pigs. The aim of this study was to investigate the associations of the p.C430S polymorphic site in exon 8 of the PPARGC1A gene with muscle fibre characteristics and meat quality traits. The polymorphism was genotyped by PCR‐RFLP using AluI restriction enzyme on a total of 152 Yorkshire pigs. Statistical analyses revealed that the p.C430S genotypes significantly affected number (P < 0.05) and area (P < 0.01) of type I muscle fibre, and were significantly associated with muscle pH (P < 0.001) and lightness (P < 0.01). On the basis of these results, we suggest that the p.C430S polymorphism can induce variation of type I fibre formation in porcine longissimus dorsi muscle and that it can be used as a meaningful molecular marker for better meat quality.
Proliferation of myofibroblasts in the stroma of renal oncocytoma
Authors: Yen T.‐H. , Chen Y. , Fu J.‐F. , Weng C.‐H. , Tian Y.‐C. , Hung C.‐C. , Lin J.‐L. , Yang C.‐W.
Published: April 2010

Objectives:  Myofibroblasts are a vital component of stroma of many malignant neoplasms, but it is not yet established whether stromal myofibroblasts also exist in benign tumours such as oncocytoma of the kidney.

Materials and methods:  Histomorphological and immunohistochemical analysis of 16 renal oncocytomas diagnosed at Chang Gung Memorial Hospital, Taiwan, has been performed.

Results:  Renal oncocytomas were composed of oncocytes, large cells with granular eosinophilic cytoplasm, arranged mostly in sheets, in tubulocystic or combined pattern. Few oncocytes appeared to be undergoing proliferation or apoptosis. MIB‐1 and active caspase 3 indices were low, but higher in tumour than in surrounding non‐tumour parenchyma (MIB‐1: 0.93 ± 0.09 versus 0.46 ± 0.07, P < 0.001 and active caspase 3: 0.76 ± 0.08 versus 0.41 ± 0.09, P < 0.001). Wnt/β‐catenin signalling was not implicated in this neoplasm, as there was no loss of E‐cadherin membranous localization or expression of intranuclear β‐catenin in the cells. Clumps of oncocytes were stained with periodic acid Schiff and had collagen I‐, collagen III‐ and fibronectin‐positive, but desmin‐ and human caldesmon‐negative stromas. Importantly, α‐smooth muscle actin (SMA)‐immunostaining established the myofibroblastic nature of many of the stromal cells. Some of the myofibroblasts were also positive for MIB‐1, indicating a proliferative role for them in the stroma.

Conclusions:  Renal oncocytomas were composed of two independent compartments: benign oncocytes and pronounced fibrotic stroma, which consisted of proliferating myofibroblasts (SMA‐ and MIB‐1‐positive) which were associated with excessive deposition of extracellular matrix (periodic acid Schiff‐component, collagen I‐, collagen III‐ and fibronectin‐positive, and desmin‐ and human caldesmon‐negative).
Renal phosphate handling in human – what can we learn from hereditary hypophosphataemias?
Authors: Amatschek Stefan , Haller Maria , Oberbauer Rainer
Published: April 2010

Eur J Clin Invest 2010; 40 (6): 552–560

Background  Renal reabsorption of inorganic phosphate is critical for the maintenance of phosphate homeostasis. The sodium dependent phosphate cotransporters NaPi‐IIa and NaPi‐IIc have been identified to fulfill this task at the brush border membrane of proximal tubule cells. Various factors including dietary phosphate intake, parathyroid hormone, or the so called phosphatonins such as FGF23 have been shown to regulate activity of these transporters.

Design  This review seeks to give an update on our current knowledge about regulatory mechanisms involved in human renal phosphate reabsorption.

Results  Recently, an increasing number of genes have been identified that are directly associated with inherited phosphate wasting disorders (Klotho, PHEX, DMP1 and NHERF1). Several of these genes are predominantly expressed by osteocytes and osteoclasts in the bone suggesting indispensable signalling pathways between kidneys and the skeleton.

Conclusion  In this review, the affected gene products in these inherited hypophosphataemias and their contribution to phosphate homeostasis are discussed.
Water retention characteristics of soils with double porosity
Authors: Russell A. R.
Published: April 2010

A soil with double porosity is modelled as a collection of aggregated particles, in which a single aggregate is made up of discrete particles bonded together. Separate fractal distributions for pore sizes around and within aggregates are defined. The particle size distribution of the double porosity soil is also modelled using a fractal distribution, which may have a fractal dimension very different to those defining the pore sizes. The surface areas of the particles and the pores within the aggregates are assumed to be equal, enabling an expression linking two fractal dimensions to be defined. It is necessary to introduce ratios between maximum and minimum particle and pore sizes into the expression. A theoretical soil‐water characteristic curve is then derived for a double porosity soil. The curve, and the underlying assumptions regarding the distributions of pore and particle sizes, showed good agreement with experimental data for a range of soils having double porosity. A discontinuity is observed in the soil‐water characteristic curve at a second air entry value related to the maximum pore size within the aggregates, a feature also observed in experimentally obtained soil‐water characteristic curves for double porosity soils.
Tyrosine phosphorylation of R3 subtype receptor‐type protein tyrosine phosphatases and their complex formations with Grb2 or Fyn
Authors: Murata Yoji , Mori Munemasa , Kotani Takenori , Supriatna Yana , Okazawa Hideki , Kusakari Shinya , Saito Yasuyuki , Ohnishi Hiroshi , Matozaki Takashi
Published: April 2010

Post‐translational modification of protein tyrosine phosphatases (PTPs) is implicated in functional modulation of these enzymes. Stomach cancer–associated protein tyrosine phosphatase‐1 (SAP‐1), as well as protein tyrosine phosphatase receptor type O (PTPRO) and vascular endothelial‐protein tyrosine phosphatase (VE‐PTP) are receptor‐type PTPs (RPTPs), which belong to the R3 subtype RPTP family. Here, we have shown that the carboxyl (COOH)‐terminal region of SAP‐1 undergoes tyrosine phosphorylation by the treatment with a PTP inhibitor. Src family kinases are important for the tyrosine phosphorylation of SAP‐1. Either Grb2 or Fyn, through their Src homology‐2 domains, bound to the tyrosine‐phosphorylated SAP‐1. Moreover, both PTPRO and VE‐PTP underwent tyrosine phosphorylation in their COOH‐terminal regions. Tyrosine phosphorylation of VE‐PTP or PTPRO also promoted their complex formations with Grb2 or Fyn. Forced expression of SAP‐1, PTPRO or VE‐PTP promoted cell spreading and lamellipodium formation of fibroblasts that expressed an activated form of Ras. In contrast, such effects of non‐tyrosine‐phosphorylated forms of these RPTPs were markedly smaller than those of wild‐type RPTPs. Our results thus suggest that tyrosine phosphorylation of R3 subtype RPTPs promotes their complex formations with Grb2 or Fyn and thus participates in the regulation of cell morphology.
Environmental controls on CH4 emission from polygonal tundra on the microsite scale in the Lena river delta, Siberia
Authors: SACHS TORSTEN , GIEBELS MICHAEL , BOIKE JULIA , KUTZBACH LARS
Published: April 2010

The carbon budgets of the atmosphere and terrestrial ecosystems are closely coupled by vertical gas exchange fluxes. Uncertainties remain with respect to high latitude ecosystems and the processes driving their temporally and spatially highly variable methane (CH4) exchange. Problems associated with scaling plot measurements to larger areas in heterogeneous environments are addressed based on intensive field studies on two nested spatial scales in Northern Siberia. CH4 fluxes on the microsite scale (0.1–100 m2) were measured in the Lena River Delta from July through September 2006 by closed chambers and were compared with simultaneous ecosystem scale (104–106 m2) flux measurements by the eddy covariance (EC) method. Closed chamber measurements were conducted almost daily on 15 plots in four differently developed polygon centers and on a polygon rim. Controls on CH4 emission were identified by stepwise multiple regression. In contrast to relatively low ecosystem‐scale fluxes controlled mainly by near‐surface turbulence, fluxes on the microsite scale were almost an order of magnitude higher at the wet polygon centers and near zero at the drier polygon rim and high‐center polygon. Microsite scale CH4 fluxes varied strongly even within the same microsites. The only statistically significant control on chamber‐based fluxes was surface temperature calculated using the Stefan–Boltzmann equation in the wet polygon centers, whereas no significant control was found for the low emissions from the dry sites. The comparison with the EC measurements reveals differences in controls and the seasonal dynamics between the two measurement scales, which may have consequences for scaling and process‐based models. Despite those differences, closed chamber measurements from within the EC footprint could be scaled by an area‐weighting approach of landcover classes based on high‐resolution imagery to match the total ecosystem‐scale emission. Our nested sampling design allowed for checking scaling results against measurements and to identify potentially missed sources or sinks.
Net primary productivity allocation and cycling of carbon along a tropical forest elevational transect in the Peruvian Andes
Authors: GIRARDIN C. A. J. , MALHI Y. , ARAGÃO L. E. O. C. , MAMANI M. , HUARACA HUASCO W. , DURAND L. , FEELEY K. J. , RAPP J. , SILVA‐ESPEJO J. E. , SILMAN M. , SALINAS N. , WHITTAKER R. J.
Published: April 2010

The net primary productivity, carbon (C) stocks and turnover rates (i.e. C dynamics) of tropical forests are an important aspect of the global C cycle. These variables have been investigated in lowland tropical forests, but they have rarely been studied in tropical montane forests (TMFs). This study examines spatial patterns of above‐ and belowground C dynamics along a transect ranging from lowland Amazonia to the high Andes in SE Peru. Fine root biomass values increased from 1.50 Mg C ha−1 at 194 m to 4.95 ± 0.62 Mg C ha−1 at 3020 m, reaching a maximum of 6.83 ± 1.13 Mg C ha−1 at the 2020 m elevation site. Aboveground biomass values decreased from 123.50 Mg C ha−1 at 194 m to 47.03 Mg C ha−1 at 3020 m. Mean annual belowground productivity was highest in the most fertile lowland plots (7.40 ± 1.00 Mg C ha−1 yr−1) and ranged between 3.43 ± 0.73 and 1.48 ± 0.40 Mg C ha−1 yr−1 in the premontane and montane plots. Mean annual aboveground productivity was estimated to vary between 9.50 ± 1.08 Mg C ha−1 yr−1 (210 m) and 2.59 ± 0.40 Mg C ha−1 yr−1 (2020 m), with consistently lower values observed in the cloud immersion zone of the montane forest. Fine root C residence time increased from 0.31 years in lowland Amazonia to 3.78 ± 0.81 years at 3020 m and stem C residence time remained constant along the elevational transect, with a mean of 54 ± 4 years. The ratio of fine root biomass to stem biomass increased significantly with increasing elevation, whereas the allocation of net primary productivity above‐ and belowground remained approximately constant at all elevations. Although net primary productivity declined in the TMF, the partitioning of productivity between the ecosystem subcomponents remained the same in lowland, premontane and montane forests.
Non‐genetic risk factors and the development of inhibitors in haemophilia: a comprehensive review and consensus report
Authors: ASTERMARK J. , ALTISENT C. , BATOROVA A. , DINIZ M. J. , GRINGERI A. , HOLME P. A. , KARAFOULIDOU A. , LOPEZ‐FERNÁNDEZ M. F. , REIPERT B. M. , ROCINO A. , SCHIAVONI M. , Von DEPKA M. , WINDYGA J. , FIJNVANDRAAT K. ,   ON BEHALF OF THE EUROPEAN HAEMOPHILIA THERAPY STANDARDISATION BOARD (EHTSB)
Published: April 2010

Summary.  The development of inhibitors to the infused factor in patients with haemophilia is a serious clinical problem. Recent evidence suggests that alongside the strong genetic contribution to inhibitor formation, there are a number of non‐genetic factors – perceived by the immune system as danger signals – which promote formation of inhibitors. This study provides a comprehensive review of clinical studies relating to these factors and also presents a survey of opinion concerning their importance and clinical influence, conducted among the members of the European Haemophilia Treatment Standardisation Board (EHTSB). Taken together, this information highlights the lack of robust data concerning the influence of several non‐genetic risk factors on inhibitor development, and an urgent need for prospective, well‐conducted studies that adhere to recommendations made by the European Medicines Agency (EMEA) for studying inhibitors. Based on current literature, the EHTSB formulated consensus recommendations. It is desirable to minimize intensive treatment wherever possible, given the clinical situation. Prophylaxis should be offered to all children, although we still need to determine optimal dosing with respect to inhibitor development, and age for starting treatment. Vaccinations should be given subcutaneously and concomitant factor concentrate infusions avoided. According to the board, there is no evidence in the literature supporting suggestions that the type of concentrate influences inhibitor risk; but all patients should be monitored during their first exposures. Furthermore, there is no evidence to support an association between pregnancy‐related issues, breast feeding and treatment‐related factors (e.g. route of administration, or use of blood components) and inhibitor development.
Crystal structure of Helicobacter pylori MinE, a cell division topological specificity factor
Authors: Kang Gil Bu , Song Hye‐Eun , Kim Mun‐Kyoung , Youn Hyung‐Seop , Lee Jung‐Gyu , An June Yop , Chun Jang‐Soo , Jeon Hyesung , Eom Soo Hyun
Published: April 2010

In Gram‐negative bacteria, proper placement of the FtsZ ring, mediated by nucleoid occlusion and the activities of the dynamic oscillating Min proteins MinC, MinD and MinE, is required for correct positioning of the cell division septum. MinE is a topological specificity factor that counters the activity of MinCD division inhibitor at the mid‐cell division site. Its structure consists of an anti‐MinCD domain and a topology specificity domain (TSD). Previous NMR analysis of truncated Escherichia coli MinE showed that the TSD domain contains a long α‐helix and two anti‐parallel β‐strands, which mediate formation of a homodimeric α/β structure. Here we report the crystal structure of full‐length Helicobacter pylori MinE and redefine its TSD based on that structure. The N‐terminal region of the TSD (residues 19–26), previously defined as part of the anti‐MinCD domain, forms a β‐strand (βA) and participates in TSD folding. In addition, H. pylori MinE forms a dimer through the interaction of anti‐parallel βA‐strands. Moreover, we observed serial dimer–dimer interactions within the crystal packing, resulting in the formation of a multimeric structure. We therefore redefine the functional domain of MinE and propose that a multimeric filamentous structure is formed through anti‐parallel β‐strand interactions.
An antisense RNA that governs the expression kinetics of a multifunctional virulence gene
Authors: Lee Eun‐Jin , Groisman Eduardo A.
Published: April 2010

Genome‐wide transcriptome analyses of several bacterial species have recently uncovered a hitherto unappreciated amount of antisense transcription. However, the physiological role, regulation and significance of such antisense transcripts are presently unclear. We now report the identification of a cis‐encoded 1.2 kb long antisense RNA – termed AmgR – that is complementary to the mgtC portion of the mgtCBR polycistronic message from Salmonella enterica. The mgtCBR mRNA specifies the MgtC protein, which is necessary for survival within macrophages, virulence in mice and growth in low Mg2+; the Mg2+ transporter MgtB with no apparent role in virulence; and the membrane peptide MgtR mediating MgtC degradation. Expression of AmgR dimished both MgtC and MgtB protein levels in a process requiring RNase E but independent of RNase III, the RNA chaperone Hfq, and the regulatory peptide MgtR. Inactivation of the chromosomal amgR promoter increased MgtC and MgtB protein levels and enhanced Salmonella virulence. Surprisingly, AmgR transcription is governed by the regulatory protein PhoP, which also directs transcription of the sense mgtCBR mRNA. AmgR may function as a timing device that alters MgtC and MgtB levels after the onset of PhoP‐inducing conditions.
Elongator function in tRNA wobble uridine modification is conserved between yeast and plants
Authors: Mehlgarten Constance , Jablonowski Daniel , Wrackmeyer Uta , Tschitschmann Susan , Sondermann David , Jäger Gunilla , Gong Zhizhong , Byström Anders S. , Schaffrath Raffael , Breunig Karin D.
Published: April 2010

Based on studies in yeast and mammalian cells the Elongator complex has been implicated in functions as diverse as histone acetylation, polarized protein trafficking and tRNA modification. Here we show that Arabidopsis mutants lacking the Elongator subunit AtELP3/ELO3 have a defect in tRNA wobble uridine modification. Moreover, we demonstrate that yeast elp3 and elp1 mutants expressing the respective Arabidopsis Elongator homologues AtELP3/ELO3 and AtELP1/ELO2 assemble integer Elongator complexes indicating a high degree of structural conservation. Surprisingly, in vivo complementation studies based on Elongator‐dependent tRNA nonsense suppression and zymocin tRNase toxin assays indicated that while AtELP1 rescued defects of a yeast elp1 mutant, the most conserved Elongator gene AtELP3, failed to complement an elp3 mutant. This lack of complementation is due to incompatibility with yeast ELP1 as coexpression of both plant genes in an elp1 elp3 yeast mutant restored Elongator's tRNA modification function in vivo. Similarly, AtELP1, not ScELP1 also supported partial complementation by yeast–plant Elp3 hybrids suggesting that AtElp1 has less stringent sequence requirements for Elp3 than ScElp1. We conclude that yeast and plant Elongator share tRNA modification roles and propose that this function might be conserved in Elongator from all eukaryotic kingdoms of life.
Molecular variability of Spanish and Hungarian isolates of Tomato torrado virus
Authors: Alfaro‐Fernández A. , Cebrián M. C. , Herrera‐Vásquez J. A. , Córdoba‐Sellés M. C. , Sánchez‐Navarro J. A. , Jordá C.
Published: April 2010

The population structure and genetic variation of Tomato torrado virus (ToTV) were estimated from 19 Spanish isolates collected from 2001 to 2009 in different tomato‐production areas by analyses of the partial nucleotide sequences of five regions of the virus genome: the protease cofactor (Pro‐Co) and the RNA‐dependent RNA polymerase (RdRp) in RNA1, and the movement protein (MP) and two subunits of the coat protein (CP; viz. Vp35 and Vp23) in RNA2. Three Hungarian isolates of the virus were also included in the analyses. All the ToTV isolates clustered together in the phylogenetic analysis of the nucleotide sequences of the different regions. However, some genetic diversity was observed in the case of the two CP subunits among the Gran Canaria isolates and the remaining ToTV‐isolates analysed, which grouped together. A high similarity was observed among all the isolates and the two published ToTV isolates: the ToTV type isolate (PRI‐ToTV0301) and the Polish isolate Wal03. The most variable encoding regions studied were those on RNA2. In general, no correlation was found between genetic diversity and collection date. Studying the genetic distances between pairs of sequences, the ratio between nonsynonymous (amino‐acid‐replacing) and synonymous (silent mutational) substitutions was low, indicating a strong negative selective pressure in the studied regions. Nine negatively selected sites (distributed in Pro‐Co, MP, Vp23 and Vp35) and just one positively selected one (in Pro‐Co) were found for all the genome regions studied.
Spatial variability of fusarium head blight pathogens and associated mycotoxins in wheat crops
Authors: Oerke E.‐C. , Meier A. , Dehne H.‐W. , Sulyok M. , Krska R. , Steiner U.
Published: April 2010

The spatial pattern of Fusarium‐infected kernels and their mycotoxin contamination was studied in four wheat fields in Germany using geo‐referenced sampling grids (12–15 × 20–30 m, 28–30 samples per field) at harvest. For each sample, frequency of Fusarium‐infected kernels and spectrum of species were assessed microbiologically; mycotoxin contents were determined by HPLC‐MS/MS analysis. Spatial variability of pathogens and mycotoxins was analysed using various parameters including Spatial Analysis by Distance IndicEs (sadie®). Microdochium majus, the most frequent head blight pathogen in 1998, was less frequent in 1999 and could not be detected in kernels from two fields in 2004. Fusarium avenaceum, F. graminearum and F. poae were the most frequent Fusarium species, with 7–8 species per field. The frequency of Fusarium‐infected kernels was 3–15% and the incidence of species showed considerable within‐field variability. Spatial patterns varied among Fusarium species as well as from field to field. Although pathogens and mycotoxin were often distributed randomly in the field, F. avenaceum, F. graminearum, F. poae, F. sporotrichioides, F. tricinctum and the mycotoxin moniliformin had an aggregated pattern in at least one field. Patterns are discussed in relation to spread of Fusarium species depending on inoculum sources, spore type, kind of dispersal, availability of susceptible host tissue and micro‐climate. Sampling of wheat fields for representative assessment of mycotoxins is complicated by random patterns of Fusarium‐infected kernels, especially where the frequency of infection is small.
Colonization of Cauliflower Blossom (Brassica oleracea) by Xanthomonas campestris pv. campestris, via Flies (Calliphora vomitoria) Can Result in Seed Infestation
Authors: Van Der Wolf Jan M , Van Der Zouwen Patricia S
Published: April 2010

Inoculation of cauliflower blossom with Xanthomonas campestris pv. campestris (Xcc), by brush or pollination with blue bottle flies (Calliphora vomitoria) as a vector, can result in seed infestation. Two years of poly‐tunnel experiments with fly inoculation of cauliflower has shown that in approximately 30% of seed batches of a breeding line, high densities of Xcc (>10 000 CFU/g) were found both before and after seed treatment with warm water. The presence of Xcc in seed derived from fly‐inoculated plants was confirmed in a grow‐out test. After brush‐inoculation, on average 45% of seed batches were contaminated with high densities of Xcc before disinfection and 40% after seed disinfection. A grow‐out test, however, did not confirm the presence of internally infected seed. In a 1‐year trial, no seed infections were found in warm water‐treated seed derived from cv. Opaal, an open pollinated cultivar. No indication of internal seed infection was found after blossom inoculation with X. c. pv. armoraciae (Xca). To obtain contaminated flies as a vector for Xcc, flies were fed on agar‐grown bacteria. Initial populations of Xcc per fly were 105 CFU, but population densities decreased rapidly and survived for 5 days maximum. During the same period, populations of other bacteria associated with flies increased slightly. The implications of blossom infection and the role of pollinating insects in dissemination and transmission of Xcc are discussed.
Distribution of microbial populations and their relationship with environmental parameters in the coastal waters of Qingdao, China
Authors: Wang Min , Liang Yantao , Bai Xiaoge , Jiang Xuejiao , Wang Fang , Qiao Qian
Published: April 2010

In order to understand the large‐scale distribution of microbial populations simultaneously and their relationship with environmental parameters, flow cytometry was used to analyse samples collected from 46 stations in the coastal waters of Qingdao in spring, 2007. The distribution of virus was significantly and positively correlated with heterotrophic bacteria. Two groups of picophytoplankton (Synechococcus and picoeukaryotes) were detected; however, Prochlorococcus was not found. Picoeukaryotes and nanophytoplankton were abundant in the near‐shore waters, whereas Synechococcus was abundant in the off‐shore areas. No variation was found in vertical distribution of virus, heterotrophic bacteria, Synechococcus and nanophytoplankton abundances, except picoeukaryotes abundance in the bottom layer was dramatically lower than that in the upper layers. Correlation analyses indicated that the relationship between abiotic variables and heterotrophic bacteria, pico‐ and nanophytoplankton was closer than that between abiotic variables and virioplankton. Temperature and nutrients were the synchronous factors controlling the growth of heterotrophic bacteria, pico‐ and nanophytoplankton in the coastal waters of Qingdao in spring. The results suggested that synergistic and antagonistic effects existed among microbial groups.
The PhaD regulator controls the simultaneous expression of the pha genes involved in polyhydroxyalkanoate metabolism and turnover in Pseudomonas putida KT2442
Authors: De Eugenio Laura Isabel , Galán Beatriz , Escapa Isabel F. , Maestro Beatriz , Sanz Jesús M. , García José Luis , Prieto María A.
Published: April 2010

The promoters of the pha gene cluster encoding the enzymes involved in the metabolism of polyhydroxyalkanoates (PHAs) in the model strain Pseudomonas putida KT2442 have been identified and compared. The pha locus is composed by five functional promoters upstream the phaC1, phaZ, phaC2, phaF and phaI genes (PC1, PZ, PC2, PF and PI respectively). PC1 and PI are the most active promoters of the pha cluster allowing the transcription of phaC1ZC2D and phaIF operons. All promoters with the sole exception of PF are carbon source‐dependent. Their transcription profiles explain the simultaneous production of PHA depolymerase and synthases to maintain the metabolic balance and PHA turnover. Mutagenesis analyses demonstrated that PhaD, a TetR‐like transcriptional regulator, behaves as a carbon source‐dependent activator of the pha cluster. The phaD gene is mainly transcribed as part of the phaC1ZC2D transcription unit and controls its own transcription and that of phaIF operon. The ability of PhaD to bind the PC1 and PI promoters was analysed by gel retardation and DNase I footprinting assays, demonstrating that PhaD interacts with a region of 25 bp at PC1 promoter (named OPRc1) and a 29 bp region at PI promoter (named OPRi). These operators contain a single binding site formed by two inverted half sites of 6 bp separated by 8 bp which overlap the corresponding promoter boxes. The 3D model structure of PhaD activator predicts that the true effector might be a CoA‐intermediate of fatty acid β‐oxidation.
Parents’ Experiences of the Provision of Community‐Based Family Support and Therapy Services Utilizing the Strengths Approach and Natural Learning Environments
Authors: Carlson Glenys , Armitstead Clare , Rodger Sylvia , Liddle Gwen
Published: April 2010

Background  In recent years, community based therapy service providers have explored different service delivery models to optimize child and family outcomes. This qualitative study aimed to explore parents’ experiences of one particular service team that adopted a strengths approach, utilizing natural learning environments.

Materials and methods  Nine parents undertook in‐depth, semi‐structured interviews that were taped, transcribed, and examined using thematic content analysis. Rigour was ensured through peer and member checking, field journals and an audit trail.

Results  Several key themes emerged including; families’ initial experiences of the service, their views of their child with a disability, their hopes for the future, and their experiences of receiving intervention from the team.

Conclusions  Parents were generally positive about the adoption of a strengths approach. ‘Working together’, ‘being positive’ and ‘information exchange’ were the main themes that parents’ attributed to their positive experiences with the service.
The ecology of saprophagous macroarthropods (millipedes, woodlice) in the context of global change
Authors: David Jean‐François , Handa Ira Tanya
Published: April 2010

Millipedes (Diplopoda) and woodlice (Crustacea, Isopoda), with a total of about 15000 described species worldwide, contribute substantially to invertebrate biodiversity. These saprophagous macroarthropods, which are key regulators of plant litter decomposition, play an important role in the functioning of terrestrial ecosystems in tropical and temperate areas. Herein we review current knowledge on the effects of climate, food quality and land cover on millipede and woodlouse species to explore their potential responses to global change. Essentially similar trends are observed in the two taxa. Experiments have shown that climate warming could result in higher rates of population growth and have positive effects on the abundance of some temperate species. This is consistent with signs of northward expansion in Europe, although the mechanisms of dispersal remain unclear. The generality of this finding is evaluated in relation to the life histories and geographical distributions of species. At low latitudes, interactions with more severe droughts are likely and could affect community composition. Elevated atmospheric CO2levels and changes in plant community composition are expected to alter leaf litter quality, a major determinant of macroarthropod fertilityviathe link with female adult body size. Although food quality changes have been shown to influence population growth rates significantly, it is proposed that the effects of warming will be probably more important during the coming decades. Land cover changes, mainly due to deforestation in the tropics and land abandonment in Europe, are critical to habitat specialists and could override any other effect of global change. Habitat destruction by man may be the main threat to macroarthropod species, many of which are narrow endemics. At the landscape scale, habitat heterogeneity could be a good option for conservation, even at the cost of some fragmentation. Two principal areas are identified which require further work: (i) the effects of climate change across broader geographic ranges, and on species with different ecologies and life histories; (ii) the effects of global change on both macroarthropods and their natural enemies (predators, parasites and pathogens), to improve predictions in field situations.
Antimicrobial peptides generated from milk proteins: a survey and prospects for application in the food industry. A review
Authors: BENKERROUM NOREDDINE
Published: April 2010

Milk proteins constitute a natural reservoir of bioactive peptides with physiological and/or antimicrobial properties, the release of which requires hydrolysis of the precursor molecules by digestive proteases or by fermentation with proteolytic micro‐organisms. Depending on the digestive or microbial proteases used, an array of bioactive peptides would be released either from caseins or whey proteins, but only a small part of these peptides has so far been identified and characterised with respect to their antimicrobial activity. The antimicrobial peptides known thus far have proven to be potent inhibitors to the growth of a wide range of undesirable micro‐organisms of health or spoilage significance. Nevertheless, previous research work has largely been oriented towards their possible application in medicine, which has hindered their high potential as food‐grade biopreservatives and/or as supplements in functional foods. This review attempts to study the literature pertaining to antimicrobial peptides derived from major milk proteins (caseins, α‐lactalbumin and β‐lactoglobulin) upon hydrolysis either by digestive proteases or by fermentation with proteolytic lactic acid bacteria. Their possible application in the food industry and their mechanism of action will also be discussed. Reference antimicrobial peptides produced by living micro‐organisms as innate immune defence components against microbial infections will occasionally be invoked for comparison purposes.