bluuebunny/crossref_metadata_2025 huggingface dataset:
Depolarization induces a conformational change in the binding site region of the M
2
muscarinic receptor
Authors: Dekel Noa , Priest Michael F. , Parnas Hanna , Parnas Itzchak , Bezanilla Francisco
Published: December 2011

G protein-coupled receptors play a central role in signal transduction and were only known to be activated by agonists. Recently it has been shown that membrane potential also affects the activity of G protein-coupled receptors. For the M
2
muscarinic receptor, it was further shown that depolarization induces charge movement. A tight correlation was found between the voltage-dependence of the charge movement and the voltage-dependence of the agonist binding. Here we examine whether depolarization-induced charge movement causes a conformational change in the M
2
receptor that may be responsible for the voltage-dependence of agonist binding. Using site-directed fluorescence labeling we show a voltage-dependent fluorescence signal, reflecting a conformational change, which correlates with the voltage-dependent charge movement. We further show that selected mutations in the orthosteric site abolish the fluorescence signal and concomitantly, the voltage-dependence of the agonist binding. Surprisingly, mutations in the allosteric site also abolished the voltage-dependence of agonist binding but did not reduce the fluorescence signal. Finally, we show that treatments, which reduced the charge movement or hindered the coupling between the charge movement and the voltage-dependent binding, also reduced the fluorescence signal. Our results demonstrate that depolarization-induced conformational changes in the orthosteric binding site underlie the voltage-dependence of agonist binding. Our results are also unique in suggesting that the allosteric site is also involved in controlling the voltage-dependent agonist binding.
Unmasking a shady mirror effect: Recognition of normal versus obscured faces
Authors: Vokey John R. , Hockley William E.
Published: December 2011

Hockley, Hemsworth, and Consoli (1999) found that following the study of normal faces, a recognition test of normal faces versus faces wearing sunglasses produced a mirror effect: The sunglasses manipulation decreased hit rates and increased false-alarm rates. The stimuli used by Hockley et al. (1999) consisted of separate poses of models wearing or not wearing sunglasses. In the current experiments, we separately manipulated same versus different depictions of individual faces and whether or not the faces were partially obscured. The results of a simulation and four experiments suggest that the test-based, mirror effect observed by Hockley et al. (1999) is actually two separable effects.
Differential Contributions of Set-Shifting and Monitoring to Dual-Task Interference
Authors: Cooper Richard P. , Karolina Wutke , Davelaar Eddy J.
Published: December 2011

It is commonly argued that complex behaviour is regulated by a number of “executive functions”, which work to coordinate the operation of disparate cognitive systems in the service of an overall goal. However, the identity, roles, and interactions of specific putative executive functions remain contentious, even within widely accepted tests of executive function. The authors present two experiments that use dual-task interference to provide further support for multiple distinct executive functions and to establish the differential contributions of those functions in two relatively complex executive tasks—random generation and the Wisconsin Card Sorting Test. Results are interpreted in terms of process models of the complex executive tasks.
A novel neuroprotective therapy for Parkinson’s disease using a viral noncoding RNA that protects mitochondrial Complex I activity
Authors: Kuan Wei-Li , Poole Emma , Fletcher Michael , Karniely Sharon , Tyers Pam , Wills Mark , Barker Roger A. , Sinclair John H.
Published: December 2011

Parkinson’s disease (PD) is a neurodegenerative disorder that results in the loss of nigrostriatal dopamine neurons. The etiology of this cell loss is unknown, but it involves abnormalities in mitochondrial function. In this study, we have demonstrated that the administration of a novel noncoding p137 RNA, derived from the human cytomegaloviral β2.7 transcript, can prevent and rescue dopaminergic cell death in vitro and in animal models of PD by protecting mitochondrial Complex I activity. Furthermore, as this p137 RNA is fused to a rabies virus glycoprotein peptide that facilitates delivery of RNA across the blood–brain barrier, such protection can be achieved through a peripheral intravenous administration of this agent after the initiation of a dopaminergic lesion. This approach has major implications for the potential treatment of PD, especially given that this novel agent could have the same protective effect on all diseased neurons affected as part of this disease process, not just the dopaminergic nigrostriatal pathway.
Endoplasmic reticulum stress regulation of the Kar2p/BiP chaperone alleviates proteotoxicity via dual degradation pathways
Authors: Hsu Chia-Ling , Prasad Rupali , Blackman Christie , Ng Davis T. W.
Published: December 2011

The unfolded protein response (UPR) monitors and maintains protein homeostasis in the endoplasmic reticulum (ER). In budding yeast, the UPR is a transcriptional regulatory pathway that is quiescent under normal conditions. Under conditions of acute ER stress, activation of UPR targets is essential for cell viability. How individual target genes contribute to stress tolerance is unclear. Uncovering these roles is hampered because most targets also play important functions in the absence of stress. To differentiate stress-specific roles from everyday functions, a single target gene was uncoupled from UPR control by eliminating its UPR-specific regulatory element. Through this approach, the UPR remains intact, aside from its inability to induce the designated target. Applying the strategy to the major ER chaperone Kar2p/BiP revealed the physiological function of increasing its cellular concentration. Despite hundreds of target genes under UPR control, we show that activation of KAR2 is indispensable to alleviate some forms of ER stress. Specifically, activation is essential to dispose misfolded proteins that are otherwise toxic. Surprisingly, induced BiP/Kar2p molecules are dedicated to alleviating stress. The inability to induce KAR2 under stress had no effect on its known housekeeping functions.
LAS1L interacts with the mammalian Rix1 complex to regulate ribosome biogenesis
Authors: Castle Christopher D. , Cassimere Erica K. , Denicourt Catherine
Published: December 2011

The coordination of RNA polymerase I transcription with pre-rRNA processing, preribosomal particle assembly, and nuclear export is a finely tuned process requiring the concerted actions of a number of accessory factors. However, the exact functions of some of these proteins and how they assemble in subcomplexes remain poorly defined. LAS1L was first described as a nucleolar protein required for maturation of the 60S preribosomal subunit. In this paper, we demonstrate that LAS1L interacts with PELP1, TEX10, and WDR18, the mammalian homologues of the budding yeast Rix1 complex, along with NOL9 and SENP3, to form a novel nucleolar complex that cofractionates with the 60S preribosomal subunit. Depletion of LAS1L-associated proteins results in a p53-dependent G1 arrest and leads to defects in processing of the pre-rRNA internal transcribed spacer 2 region. We further show that the nucleolar localization of this complex requires active RNA polymerase I transcription and the small ubiquitin-like modifier–specific protease SENP3. Taken together, our data identify a novel mammalian complex required for 60S ribosomal subunit synthesis, providing further insight into the intricate, yet poorly described, process of ribosome biogenesis in higher eukaryotes.
A potentially common peptide target in secreted heat shock protein-90α for hypoxia-inducible factor-1α–positive tumors
Authors: Sahu Divya , Zhao Zhengwei , Tsen Fred , Cheng Chieh-Fang , Park Ryan , Situ Alan J. , Dai Jinyao , Eginli Ariana , Shams Sharmineh , Chen Mei , Ulmer Tobias S. , Conti Peter , Woodley David T. , Li Wei
Published: December 2011

Deregulated accumulation of hypoxia-inducible factor-1α (HIF-1α) is a hallmark of many solid tumors. Directly targeting HIF-1α for therapeutics is challenging. Our finding that HIF-1α regulates secretion of heat shock protein-90α (Hsp90α) for cell migration raises the exciting possibility that targeting the secreted Hsp90α from HIF-1α–positive tumors has a better clinical outlook. Using the HIF-1α–positive and metastatic breast cancer cells MDA-MB-231, we show that down-regulation of the deregulated HIF-1α blocks Hsp90α secretion and invasion of the cells. Reintroducing an active, but not an inactive, HIF-1α into endogenous HIF-1α–depleted cells rescues both Hsp90α secretion and invasion. Inhibition of Hsp90α secretion, neutralization of secreted Hsp90α action, or removal of the cell surface LRP-1 receptor for secreted Hsp90α reduces the tumor cell invasion in vitro and lung colonization and tumor formation in nude mice. Furthermore, we localized the tumor-promoting effect to a 115–amino acid region in secreted Hsp90α called F-5. Supplementation with F-5 is sufficient to bypass the blockade of HIF-1α depletion and resumes invasion by the tumor cells under serum-free conditions. Because normal cells do not secrete Hsp90α in the absence of stress, drugs that target F-5 should be more effective and less toxic in treatment of HIF-1α–positive tumors in humans.
Aortic arch enlargement and coarctation repair through a left thoracotomy: significance of ductal perfusion
Authors: Dave Hitendu , Rosser Barbara , Reineke Kim , Nguyen-Minh Sylvie , Knirsch Walter , Prêtre René
Published: December 2011

OBJECTIVE

To analyse the technique of neonatal aortic arch enlargement without cardiopulmonary bypass through a left posterior thoracotomy, as an adjunct to extended resection for Coarctation and severe arch hypoplasia.

METHODS

Ten neonates with coarctation, severe arch hypoplasia and a persistent ductus arteriosus (PDA) were subjected to arch repair through a left posterior thoracotomy. Nine of these patients had associated significant intracardiac anomalies; three of them received pulmonary artery (PA) banding. After exclusion from circulation, the roof of the intervening arch between left carotid and left subclavian was enlarged using a patch. After adequate reperfusion, a classic resection and extended end-to-end anastomosis was performed. Median age and weight were 5.5 (1–10) days and 3.3 (2.2–4.1) kg respectively. The median preoperative arch diameter was 1.07 (0.75–1.32) mm/kg body weight.

RESULTS

All patients could be successfully operated with this approach. The non-ischaemic and ischaemic aortic clamp times were 40 (15–68) and 23 (18–32) min, respectively. The median postoperative arch diameter achieved was 1.43 (1.06–1.46) mm/kg body weight. None of the patients had significant gradient early postoperatively. Two patients with recurrent stenosis were successfully treated with balloon dilatation (1) or surgery with cardiopulmonary bypass (CPB) (1). One patient has a corrected gradient of 16 mmHg in the proximal arch which is being observed. The remaining patients are free from stenosis at a median follow-up of 30.1 (13.2–57.8) months.

CONCLUSIONS

Use of PDA for lower body perfusion allows complex reconstruction of the arch without incurring lower body ischaemia. The extended resection could then be performed without excessive stretch. This modification saves these patients from undergoing a complex arch reconstruction with CPB in the early neonatal period.
Transmission of Citrus leprosis virus C by Brevipalpus phoenicis (Geijskes) to Alternative Host Plants Found in Citrus Orchards
Authors: Nunes M. A. , de Oliveira C. A. L. , de Oliveira M. L. , Kitajima E. W. , Hilf M. E. , Gottwald T. R. , Freitas-Astúa J.
Published: December 2011

The equivalent of US$75 million is spent each year in Brazil to control Brevipalpus phoenicis, a mite vector of Citrus leprosis virus C (CiLV-C). In this study, we investigated the possibility that hedgerows and windbreaks normally found in citrus orchards could host CiLV-C. Mites confined by an adhesive barrier were reared on sweet orange fruit with leprosis symptoms then were transferred to leaves of Hibiscus rosa-sinensis, Malvaviscus arboreus, Grevilea robusta, Bixa orellana, and Citrus sinensis. Ninety days post infestation, the descendant mites were transferred to Pera sweet orange plants to verify the transmissibility of the virus back to citrus. Nonviruliferous mites which had no feeding access to diseased tissue were used as controls. Local chlorotic or necrotic spots and ringspots, symptoms of leprosis disease, appeared in most plants tested. Results generated by reversetranscription polymerase chain reaction with primers specific for CiLV-C and by electron microscope analyses confirmed the susceptibility of these plants to CiLV-C.
Strain Characterization of Potato virus S Isolates from Tasmania, Australia
Authors: Lambert Susan J. , Scott Jason B. , Pethybridge Sarah J. , Hay Frank S.
Published: December 2011

Potato virus S (PVS) is prevalent within potato (Solanum tuberosum) production worldwide. Traditionally, PVS has been split into two strains, Ordinary (PVSO) and Andean (PVSA), based on reaction in herbaceous indicator species such as Chenopodium quinoa. However, recent research has identified further strain designations, such as PVSO-CS (Ordinary and Chenopodium systemic). Forty-four isolates of PVS were collected from potato seed lines in different geographical regions within Tasmania, Australia. Isolates were initially characterized by reactions in C. quinoa. Nineteen isolates were characterized as PVSO, based on the development of local lesions and serological detection in inoculated leaves only. Three isolates were identified as PVSA-like, based on local lesion development in inoculated leaves, mild mottling or chlorotic spots on noninoculated leaves, and serological detection in both inoculated and noninoculated leaves. Thirteen isolates produced no symptoms, and were detected serologically in inoculated leaves only (PVSO-like). Four isolates produced no symptoms but were detected serologically in both inoculated and noninoculated leaves (PVSA-like). Five isolates produced symptoms in inoculated leaves only but were detected serologically in both inoculated and noninoculated leaves (also PVSA-like). The ability of isolates to infect tomato has also been used as a criterion to assist in PVS strain differentiation. A subsample of isolates (n = 16) was unable to infect tomato ‘Grosse Lisse’. Seventeen isolates representative of these groupings based on reactions in C. quinoa were also characterized by coat-protein sequencing. Phylogenetic comparisons suggested that all isolates were PVSO rather than PVSA. Therefore, whereas some of these PVS isolates were systemic in C. quinoa, findings from this study suggest that they were not PVSA, and that only PVSO and PVSO-CS isolates are present in Tasmania. The implications of this finding for disease management are discussed.
Production, Harvest, and Concentration of Feline Immunodeficiency Virus-Based Lentiviral Vector from Cells Grown in CF10 or CF2 Devices
Authors: Saenz Dyana T. , Barraza Román , Loewen Nils , Teo Wulin , Poeschla Eric M.
Published: December 2011

Feline immunodeficiency virus (FIV)-based lentiviral vectors are useful for introducing integrated transgenes into nondividing human cells. This protocol describes the production of FIV-based lentiviral vectors using cells grown in CF10 or CF2 devices. It also details the harvesting and concentration of these vectors. The methods are for production for basic science laboratory use and in vivo experimentation. They do not meet standards for clinical-grade (good manufacturing practice [GMP]) production.
Single-Molecule Studies Using Magnetic Traps
Authors: Lionnet Timothée , Allemand Jean-François , Revyakin Andrey , Strick Terence R. , Saleh Omar A. , Bensimon David , Croquette Vincent
Published: December 2011

In recent years, techniques have been developed to study and manipulate single molecules of DNA and other biopolymers. In one such technique, the magnetic trap, a single DNA molecule is bound at one end to a glass surface and at the other to a magnetic microbead. Small magnets, whose position and rotation can be controlled, pull on and rotate the microbead. This provides a simple method to stretch and twist the molecule. The system allows one to apply and measure forces ranging from 10−3 to >100 pN. In contrast to other techniques, the force measurement is absolute and does not require calibration of the sensor. In this article, we describe the principle of the magnetic trap, as well as its use in the measurement of the elastic properties of DNA and the study of DNA–protein interactions.
Epidemiologic Considerations in Complex Disease Genetics: Figure 1.
Authors: Gallacher John
Published: December 2011

Association studies are becoming an increasingly popular study design not only for testing of candidate genes, but also for genome-wide localization of loci influencing complex disease. This article covers some of the practical concerns in conducting large-scale association studies using epidemiologic samples.
Analysis of Glial Cell Development and Function in Drosophila
Authors: Stork Tobias , Bernardos Rebecca , Freeman Marc R.
Published: December 2011

Glial cells are the most abundant cell type in our brains, yet we understand very little about their development and function. An accumulating body of work over the last decade has revealed that glia are critical regulators of nervous system development, function, and health. Based on morphological and molecular criteria, glia in Drosophila melanogaster are very similar to their mammalian counterparts, suggesting that a detailed investigation of fly glia has the potential to add greatly to our understanding of fundamental aspects of glial cell biology. In this article, we provide an overview of the subtypes of glial cells found in Drosophila and discuss our current understanding of their functions, the development of a subset of well-defined glial lineages, and the molecular-genetic tools available for manipulating glial subtypes in vivo.
CCA1 and ELF3 Interact in the Control of Hypocotyl Length and Flowering Time in Arabidopsis
Authors: Lu Sheen X. , Webb Candace J. , Knowles Stephen M. , Kim Sally H.J. , Wang Zhiyong , Tobin Elaine M.
Published: December 2011

The circadian clock is an endogenous oscillator with a period of approximately 24 h that allows organisms to anticipate, and respond to, changes in the environment. In Arabidopsis (Arabidopsis thaliana), the circadian clock regulates a wide variety of physiological processes, including hypocotyl elongation and flowering time. CIRCADIAN CLOCK ASSOCIATED1 (CCA1) is a central clock component, and CCA1 overexpression causes circadian dysfunction, elongated hypocotyls, and late flowering. EARLY FLOWERING3 (ELF3) modulates light input to the clock and is also postulated to be part of the clock mechanism. elf3 mutations cause light-dependent arrhythmicity, elongated hypocotyls, and early flowering. Although both genes affect similar processes, their relationship is not clear. Here, we show that CCA1 represses ELF3 by associating with its promoter, completing a CCA1-ELF3 negative feedback loop that places ELF3 within the oscillator. We also show that ELF3 acts downstream of CCA1, mediating the repression of PHYTOCHROME-INTERACTING FACTOR4 (PIF4) and PIF5 in the control of hypocotyl elongation. In the regulation of flowering, our findings show that ELF3 and CCA1 either cooperate or act in parallel through the CONSTANS/FLOWERING LOCUS T pathway. In addition, we show that CCA1 represses GIGANTEA and SUPPRESSOR OF CONSTANS1 by direct interaction with their promoters, revealing additional connections between the circadian clock and the flowering pathways.
Seasonal Abscisic Acid Signal and a Basic Leucine Zipper Transcription Factor, DkbZIP5, Regulate Proanthocyanidin Biosynthesis in Persimmon Fruit
Authors: Akagi Takashi , Katayama-Ikegami Ayako , Kobayashi Shozo , Sato Akihiko , Kono Atsushi , Yonemori Keizo
Published: December 2011

Proanthocyanidins (PAs) are secondary metabolites that contribute to plant protection and crop quality. Persimmon (Diospyros kaki) has a unique characteristic of accumulating large amounts of PAs, particularly in its fruit. Normal astringent-type and mutant nonastringent-type fruits show different PA accumulation patterns depending on the seasonal expression patterns of DkMyb4, which is a Myb transcription factor (TF) regulating many PA pathway genes in persimmon. In this study, attempts were made to identify the factors involved in DkMyb4 expression and the resultant PA accumulation in persimmon fruit. Treatment with abscisic acid (ABA) and an ABA biosynthesis inhibitor resulted in differential changes in the expression patterns of DkMyb4 and PA biosynthesis in astringent-type and nonastringent-type fruits depending on the development stage. To obtain an ABA-signaling TF, we isolated a full-length basic leucine zipper (bZIP) TF, DkbZIP5, which is highly expressed in persimmon fruit. We also showed that ectopic DkbZIP5 overexpression in persimmon calluses induced the up-regulation of DkMyb4 and the resultant PA biosynthesis. In addition, a detailed molecular characterization using the electrophoretic mobility shift assay and transient reporter assay indicated that DkbZIP5 recognized ABA-responsive elements in the promoter region of DkMyb4 and acted as a direct regulator of DkMyb4 in an ABA-dependent manner. These results suggest that ABA signals may be involved in PA biosynthesis in persimmon fruit via DkMyb4 activation by DkbZIP5.
Influence of chloroform on crystalline products yielded in reactions of 5,10,15,20-tetraphenylporphyrin with HCl and copper(II) salts
Authors: Aparici Plaza Luis , Chojnacki Jaroslaw
Published: December 2011

Chloroform was found to occupy the lattice of the protonated porphyrin and to promote crystallization of a different polymorphic form of a metalloporphyrin. The structure of 5,10,15,20-tetraphenylporphyrin-21,23-diium dichloride chloroform octasolvate, C44H32N42+·2Cl−·8CHCl3, (I), in the solid state is described and compared with related solvates. The porphyrin macrocycle displays a distorted saddle shape, with chloride anions above and below the ring. Seven chloroform molecules are boundviaC—H...Cl hydrogen bonds, while the link with the eighth solvent molecule is weaker. A new monoclinic polymorph of (5,10,15,20-tetraphenylporphyrinato)copper(II), [Cu(C44H28N4)], (II), crystallized from chloroform, is also presented.
A review of medication incidents reported to the National Reporting and Learning System in England and Wales over 6 years (2005–2010)
Authors: Cousins David H. , Gerrett David , Warner Bruce
Published: December 2011

A review of all medication incidents reported to the National Reporting and Learning System (NRLS) in England in Wales between 1 January 2005 and 31 December 2010 was undertaken. The 526 186 medication incident reports represented 9.68% of all patient safety incidents. Medication incidents from acute general hospitals (394 951) represented 75% of reports. There were relatively smaller numbers of medication incident reports (44 952) from primary care, representing 8.5% of the total. Of 86 821 (16%) medication incidents reporting actual patient harm, 822 (0.9%) resulted in death or severe harm. The incidents involving medicine administration (263 228; 50%) and prescribing (97 097; 18%) were the process steps with the largest number of reports. Omitted and delayed medicine (82 028; 16%) and wrong dose (80 170; 15%) represented the largest error categories. Thirteen medicines or therapeutic groups accounted for 377 (46%) of the incidents with outcomes of death or severe harm. The National Patient Safety Agency (NPSA) has issued guidance to help minimize incidents with many of these medicines. Many recent incidents could have been prevented if the NPSA guidance had been better implemented. It is recommended that healthcare organizations in all sectors establish an effective infrastructure to oversee and promote safe medication practice, including an annual medication safety report. In the future, preventable harms from medication incidents can be further minimized by; the continued use of the NRLS to identify and prioritize important actions to improve medication safety, a central organization continuing to issue medication safety guidance to the service and better methods to ensure that the National Health Service has implemented this guidance.
Is epitope recognition of shrimp allergens useful to predict clinical reactivity?
Authors: Ayuso R. , Sánchez‐Garcia S. , Pascal M. , Lin J. , Grishina G. , Fu Z. , Ibáñez M. D. , Sastre J. , Sampson H. A.
Published: December 2011

Background

Shrimp is a frequent cause of severe allergic reactions world‐wide. Due to issues such as cross‐reactivity, diagnosis of shrimp allergy is still inaccurate, requiring the need for double‐blind, placebo‐controlled food challenges (DBPCFC). A better understanding of the relationship between laboratory findings and clinical reactivity is needed.

Objective

To determine whether sensitization to certain shrimp allergens or recognition of particular IgE epitopes of those allergens are good biomarkers of clinical reactivity to shrimp.

Methods

Thirty‐seven consecutive patients were selected with clinical histories of shrimp allergy. Skin prick test, specific IgE determinations, DBPCFC and immunoblot assays to shrimp extract were performed. IgE binding to synthetic overlapping peptides representing the sequence of the four allergens from the Pacific white shrimp (Litopenaeus vannamei) identified to date (Lit v1, Lit v2, Lit v3 and Lit v4) was analysed.

Results

Of 37 (46%) patients, 17 had a positive challenge to shrimp (11 children and 6 adults). By microarray, patients with positive challenges showed more intense binding to shrimp peptides than those with negative challenges. Statistically significant differences in terms of the frequency and intensity of IgE binding to some epitopes were observed between the two groups. Diagnostic efficiency was higher for individual epitopes than for proteins. Particularly, efficiency was highest for certain Lit v 1 and Lit v 2 epitopes, followed by Lit v 3 and Lit v 4 epitopes.

Conclusion and Clinical Relevance

Patients with positive shrimp challenges present in general more intense and diverse epitope recognition to all four shrimp allergens. IgE antibodies to these shrimp epitopes could be used as biomarkers for prediction of clinical reactivity in subjects with sensitization to shrimp. Patients with positive shrimp challenges show more intense sensitization and more diverse epitope recognition. Several IgE‐binding shrimp epitopes could be used as biomarkers for predicting clinical reactivity in subjects with sensitization to shrimp.
T‐cell receptor affinity in thymic development
Authors: Moran Amy E. , Hogquist Kristin A.
Published: December 2011

Understanding the thymic processes that support the generation of functionally competent and self‐tolerant lymphocytes requires dissection of the T‐cell receptor (TCR) response to ligands of different affinities. In spatially segregated regions of the thymus, with unique expression of proteases and cytokines, TCR affinity guides a number of cell fate decisions. Yet affinity alone does not explain the selection paradox. Increasing evidence suggests that the ‘altered peptide’ model of the 1980s together with the affinity model might best explain how the thymus supports conventional and regulatory T‐cell development. Development of new tools to study the strength of TCR signals perceived by T cells, novel regulatory T‐cell transgenic mice, and tetramer enrichment strategies have provided an insight into the nature of TCR signals perceived during thymocyte development. These topics are discussed and support for the prevailing hypotheses is presented.
BiodentineTM induces TGF‐β1 release from human pulp cells and early dental pulp mineralization
Authors: Laurent P. , Camps J. , About I.
Published: December 2011

Laurent P, Camps J, About I. Biodentine™ induces TGF‐β1 release from human pulp cells and early dental pulp mineralization. International Endodontic Journal, 45, 439–448, 2012.

Aim  To assess the ability of a recently developed tricalcium silicate‐based cement (Biodentine™) to induce reparative dentine synthesis and to investigate its capacity to modulate pulp cells TGF‐β1 secretion.

Methodology  Biodentine™ was directly applied onto the dental pulp in an entire human tooth culture model. After various culture periods, the interaction of the material with dental pulp tissue was analysed on tissue sections. The effect of increasing surface area of this material on TGF‐β1 secretion was investigated on pulp cell cultures and compared with that of MTA, calcium hydroxide and Xeno®III adhesive resin. After performing artificial injuries on pulp cell cultures, the materials eluates were added for 24 h and then TGF‐β1 secretion was quantified by ELISA. Controls were performed by incubating intact cells with the culture medium, while injured cells TGF‐β1 level was used as the baseline value.

Results  Biodentine™ induced mineralized foci formation early after its application. The mineralization appeared under the form of osteodentine and expressed markers of odontoblasts. Biodentine™ significantly increased TGF‐β1 secretion from pulp cells (P < 0.03) independently of the contact surface increase. This increase was also observed with calcium hydroxide and MTA, but not with the resinous Xeno®III. The statistical analysis showed statistically significant differences between capping materials and the resinous Xeno®III (P < 0.001).

Conclusions  When Biodentine™ was applied directly onto the pulp, it induced an early form of reparative dentine synthesis, probably due to a modulation of pulp cell TGF‐β1 secretion.
Towards novel approaches to modelling biotic interactions in multispecies assemblages at large spatial extents
Authors: Kissling W. D. , Dormann Carsten F. , Groeneveld Jürgen , Hickler Thomas , Kühn Ingolf , McInerny Greg J. , Montoya José M. , Römermann Christine , Schiffers Katja , Schurr Frank M. , Singer Alexander , Svenning Jens‐Christian , Zimmermann Niklaus E. , O’Hara Robert B.
Published: December 2011

Aim  Biotic interactions – within guilds or across trophic levels – have widely been ignored in species distribution models (SDMs). This synthesis outlines the development of ‘species interaction distribution models’ (SIDMs), which aim to incorporate multispecies interactions at large spatial extents using interaction matrices.

Location  Local to global.

Methods  We review recent approaches for extending classical SDMs to incorporate biotic interactions, and identify some methodological and conceptual limitations. To illustrate possible directions for conceptual advancement we explore three principal ways of modelling multispecies interactions using interaction matrices: simple qualitative linkages between species, quantitative interaction coefficients reflecting interaction strengths, and interactions mediated by interaction currencies. We explain methodological advancements for static interaction data and multispecies time series, and outline methods to reduce complexity when modelling multispecies interactions.

Results  Classical SDMs ignore biotic interactions and recent SDM extensions only include the unidirectional influence of one or a few species. However, novel methods using error matrices in multivariate regression models allow interactions between multiple species to be modelled explicitly with spatial co‐occurrence data. If time series are available, multivariate versions of population dynamic models can be applied that account for the effects and relative importance of species interactions and environmental drivers. These methods need to be extended by incorporating the non‐stationarity in interaction coefficients across space and time, and are challenged by the limited empirical knowledge on spatio‐temporal variation in the existence and strength of species interactions. Model complexity may be reduced by: (1) using prior ecological knowledge to set a subset of interaction coefficients to zero, (2) modelling guilds and functional groups rather than individual species, and (3) modelling interaction currencies and species’ effect and response traits.

Main conclusions  There is great potential for developing novel approaches that incorporate multispecies interactions into the projection of species distributions and community structure at large spatial extents. Progress can be made by: (1) developing statistical models with interaction matrices for multispecies co‐occurrence datasets across large‐scale environmental gradients, (2) testing the potential and limitations of methods for complexity reduction, and (3) sampling and monitoring comprehensive spatio‐temporal data on biotic interactions in multispecies communities.
A new timeframe for the diversification of Japan’s mammals
Authors: McKay Bailey D.
Published: December 2011

Aim The main Japanese islands are land‐bridge islands divided by the biogeographic division Blakiston’s Line and represent two natural laboratories for studying land‐bridge diversification. Colonization of the current mammal fauna has been dated to the middle to late Pleistocene using fossil evidence. The purpose of this paper is to apply a molecular clock to the genetic divergences between Japanese mammalian taxa and their sister mainland taxa to test the late Pleistocene land‐bridge colonization hypothesis.

Location The main Japanese islands (Kyushu, Shikoku, Honshu and Hokkaido).

Methods I used mitochondrial DNA (cytochromeb) and a species tree approach to estimate the divergence times of 24 Japanese non‐volant terrestrial mammal taxa and their mainland sister taxa using the program *beast. I then tested for evidence of non‐simultaneous divergence among these taxon‐pairs by controlling for expected coalescent stochasticity using the program MsBayes.

Results Divergence events between taxa on Japan and their mainland sister taxa were significantly older than expected under the current paradigm, which is based on fossil data. Consistent with the land‐bridge colonization hypothesis, there was evidence of multiple divergence events.

Main conclusions These results implicate a colonization timeframe that is older than posited by the current paradigm based on fossil evidence. However, these results are still consistent with the land‐bridge colonization hypothesis. Multiple periods of land‐bridge connectivity may account for the current mammalian fauna in Japan. In addition, half of the divergence time estimates in the Honshu–Shikoku–Kyushu region were clumped around 2.4 Ma, which might suggest a dramatic interchange period, concordant with a period of significant global cooling, when the first land bridge may have connected Japan to the mainland.
High Fat Diet Affects Reproductive Functions in Female Diet‐Induced Obese and Dietary Resistant Rats
Authors: Balasubramanian P. , Jagannathan L. , Mahaley Rochell E , Subramanian M. , Gilbreath E. T. , MohanKumar P. S. , MohanKumar S. M. J.
Published: December 2011

The incidence of ovulatory disorders is common in obese animal models. The mechanism behind this effect is unclear. We hypothesised that a high‐fat (HF) diet induces alterations in neuroendocrine mechanisms resulting in anovulation in diet‐induced obese (DIO) animals. Adult female DIO and diet‐resistant (DR) rats were fed either chow or a HF diet (45% calories from fat) for 6 weeks. Oestrous cyclicity and body weight were monitored regularly. At the end of treatment, rats were implanted with a jugular catheter to monitor luteinising hormone (LH) levels on the day of pro‐oestrous. Rats were sacrificed on the next pro‐oestrous, and their brains and ovaries were collected. Plasma from trunk blood was analysed for oestradiol and leptin concentrations. Ovaries were fixed and sectioned for histological analysis. Brains were removed, frozen and sectioned, and norepinephrine (NE) concentrations in discrete hypothalamic areas were measured using high‐performance liquid chromatography with electrochemical detection. A HF diet exposure affected oestrous cyclicity in both DIO and DR rats, with the effect being more pronounced in DIO animals. HF diet exposure increased leptin levels in both DIO and DR rats. Oestradiol levels were low in the DIO‐HF group. NE levels in the hypothalamus were unaffected by HF diet or genotype. A normal LH surge was observed in DR‐Chow rats and LH levels were low in the remaining groups. These results lead to the conclusion that DIO rats have an inherently reduced reproductive capacity and exposure to a HF diet decreases it further. A reduction in oestradiol and LH surge levels could contribute to this effect; however, the underlying mechanisms need to be investigated further.
Exercise using tongue‐holding swallow does not improve swallowing function in normal subjects
Authors: OH J.‐C. , PARK J.‐W. , CHA T.‐H. , WOO H.‐S. , KIM D.‐K.
Published: December 2011

Summary  We evaluated the effects of repetitive tongue‐holding swallow (THS) as an exercise for improving swallowing function. Twenty healthy subjects were randomly divided into two groups. One group [THS group (THSG)] performed the tongue‐holding manoeuvre every 5 s for 20 min. The other group (normal swallow group) swallowed without tongue‐holding for the same length of time as a control. Twenty sessions of training were performed for 4 weeks. All participants also had a videofluoroscopic swallowing study for analysing the biomechanical parameters of swallowing (hyolaryngeal movement, posterior pharyngeal wall movement and the pharyngeal constriction ratio). After 4 weeks of training, none of the biomechanical parameters changed in either group. Exercise using THS did not affect swallowing function in normal subjects.
Spore formation in Myxococcus xanthus is tied to cytoskeleton functions and polysaccharide spore coat deposition
Authors: Müller Frank D. , Schink Christian W. , Hoiczyk Egbert , Cserti Emöke , Higgs Penelope I.
Published: December 2011

Myxococcus xanthus is a Gram‐negative bacterium that differentiates into environmentally resistant spores. Spore differentiation involves septation‐independent remodelling of the rod‐shaped vegetative cell into a spherical spore and deposition of a thick and compact spore coat outside of the outer membrane. Our analyses suggest that spore coat polysaccharides are exported to the cell surface by the Exo outer membrane polysaccharide export/polysaccharide co‐polymerase 2a (OPX/PCP‐2a) machinery. Conversion of the capsule‐like polysaccharide layer into a compact spore coat layer requires the Nfs proteins which likely form a complex in the cell envelope. Mutants in either nfs, exo or two other genetic loci encoding homologues of polysaccharide synthesis enzymes fail to complete morphogenesis from rods to spherical spores and instead produce a transient state of deformed cell morphology before reversion into typical rods. We additionally provide evidence that the cell cytoskeletal protein, MreB, plays an important role in rod to spore morphogenesis and for spore outgrowth. These studies provide evidence that this novel Gram‐negative differentiation process is tied to cytoskeleton functions and polysaccharide spore coat deposition.
Dynamics of the assembly of a complex macromolecular structure – the coat of spores of the bacterium Bacillus subtilis
Authors: Setlow Peter
Published: December 2011

The coat is the outermost layer of spores of many Bacillus species, and plays a key role in these spores' resistance. The Bacillus subtilis spore coat contains > 70 proteins in four distinct layers: the basement layer, inner coat, outer coat and crust. In this issue of Molecular Microbiology, McKenney and Eichenberger study the dynamics of spore coat assembly using GFP‐fusions to 41 B. subtilis coat proteins. A key finding in the work is that formation of the spore coat is initiated by the apparently simultaneous assembly of foci of proteins from all four coat layers on the developing spore just as forespore engulfment by the mother cell begins. The expansion of these foci before completion of forespore engulfment then sets up the scaffold to which coat proteins added later in sporulation are added. This study provides new understanding of the mechanism of the assembly of a multi‐protein, multi‐lamellar structure.
Volatiles of two growth‐inhibiting rhizobacteria commonly engage AtWRKY18 function
Authors: Wenke Katrin , Wanke Dierk , Kilian Joachim , Berendzen Kenneth , Harter Klaus , Piechulla Birgit
Published: December 2011

Interactions with the (a)biotic environment play key roles in a plant’s fitness and vitality. In addition to direct surface‐to‐surface contact, volatile chemicals can also affect the physiology of organism. Volatiles of Serratia plymuthica and Stenotrophomonas maltophilia significantly inhibited growth and induced H2O2 production in Arabidopsis in dual culture. Within 1 day, transcriptional changes were observed by promoter–GUS assays using a stress‐inducible W‐box‐containing 4xGST1 construct. Expression studies performed at 6, 12 and 24 h revealed altered transcript levels for 889 genes and 655 genes in response to Se. plymuthica or St. maltophilia volatiles, respectively. Expression of 162 genes was altered in both treatments. Meta‐analysis revealed that specifically volatile‐responsive genes were significantly overlapping with those affected by abiotic stress. We use the term mVAMP (microbial volatile‐associated molecular pattern) to describe these volatile‐specific responses. Genes responsive to both treatments were enriched for W‐box motifs in their promoters, and were significantly enriched for transcription factors (ERF2, ZAT10, MYB73 and WRKY18). The susceptibility of wrky18 mutant lines to volatiles was significantly delayed, suggesting an indispensable role for WRKY18 in bacterial volatile responses.
Haemolysis and sublethal injury of RBCs after routine blood bank manipulations
Authors: Harm S. K. , Raval J. S. , Cramer J. , Waters J. H. , Yazer M. H.
Published: December 2011

Objectives: To determine the extent of RBC sublethal injury in male donor units as measured by both the mechanical fragility index (MFI) and percentage haemolysis after RBCs underwent leucoreduction (LR), irradiation (IRRAD), and washing.

Background: RBCs frequently undergo post‐collection processing to meet certain recipient's special needs. The extent of haemolysis and sublethal injury following these interventions has not been fully characterised.

Methods: Eight to ten day old male, AS‐5 RBCs underwent either LR, IRRAD or washing. A control group of male, AS‐5 RBCs were unmanipulated. The MFI, percent haemolysis, and plasma free haemoglobin (PFHb) were measured immediately after manipulation and, for a series of irradiated RBCs, 28 days after irradiation (IRRAD28).

Results: The MFI of the washed units was significantly higher than unmanipulated, LR, IRRAD, IRRAD28 units (P < 0·0001). The percent haemolysis was highest in the IRRAD28 units (1·4%) followed by the washed units (0·74%); the other three units demonstrated significantly less haemolysis (P < 0·0001). The largest mean total amount of PFHb per unit was found in the IRRAD28 units (500·5 mg/unit) followed by the washed units (149·8 mg/unit); the mean total amount of PFHb in the three other types of units was significantly less than that found in both the IRRAD28 and washed units (P at least < 0·001).

Conclusion: There is a significant quantity of PFHb in IRRAD28 RBC units, and potentially in washed allogeneic RBC units. Clinical correlation is required to determine if this quantity of PFHb and the transfusion of potentially fragile RBCs causes adverse events.
Fluorescence diagnostics as a guide for demarcation and biopsy of suspected anal cancer
Authors: Kriz Martin , Hegyi Juraj , Ruzicka Thomas , Berking Carola
Published: December 2011

Background  Malignant epithelial tumors of the perianal skin are relatively rare however show high recurrence rates. A preoperative precise diagnosis and demarcation of the involved cancerous skin remains challenging but is worthwhile to minimize tissue defects and possible limitations of quality of life.

Objectives  To show the use of fluorescence diagnostics (FD) in the management of chronic perianal disease.

Methods  FD was performed using methylaminolevulinate (MAL) for three hours topically followed by visualization with blue light (405 nm).

Results  We report on two patients in whom FD helped to demarcate the involved tissue or to direct biopsy to ascertain the final diagnosis. In one patient with a history of Bowen′s disease and recurrent anal carcinoma, biopsy showed extramammary Paget’s disease and thus was treated by surgical anoplasty. The other patient had a history of chronic anal eczema, and FD‐directed biopsy could exclude suspected malignancy so that the anal eczema was treated by local therapy.

Conclusions  FD is a non‐invasive technique which can be used in the perianal region in order to delineate pathologic processes which may reduce the number of surgical interventions.
The implementation of a perioperative checklist increases patients' perioperative safety and staff satisfaction
Authors: BÖHMER A. B. , WAPPLER F. , TINSCHMANN T. , KINDERMANN P. , RIXEN D. , BELLENDIR M. , SCHWANKE U. , BOUILLON B. , GERBERSHAGEN M. U.
Published: December 2011

Background

The implementation of the ‘Surgical Safety Checklist’ caused a significant reduction in the incidence of complications and mortality among patients undergoing surgery. The aim of the present study was to evaluate perioperative safety standards and the quality of interprofessional cooperation before and after the introduction of a safety checklist from staff members' point of view.

Methods

Employees' attitude concerning safety‐relevant aspects of the perioperative period, work processes, and quality of interprofessional cooperation was surveyed before and 3 months after introducing an adapted form of the ‘Surgical Safety Checklist’ by a 19‐item questionnaire.

Results

After the implementation of the checklist, the cognizance of the names and functions of the individual operating room (OR) staff members, verification of the patient's written consent for surgery, indication for antibiotics before the surgical incision, and the quality of interprofessional cooperation were rated more positively. Traumatology physicians were more convinced that all artifacts had been removed from the surgical field. Finally, communication about intraoperative complications had improved.

Conclusions

Our attitude surveys demonstrate that from the OR staff's perspective, in the perioperative setting, safety‐relevant factors can be handled significantly better and with greater awareness by implementing a safety checklist as proposed by the World Health Organization.