bluuebunny/crossref_metadata_2025 huggingface dataset:
The role of strigolactones and ethylene in disease caused by Pythium irregulare
Authors: Blake Sara N. , Barry Karen M. , Gill Warwick M. , Reid James B. , Foo Eloise
Published: September 2015

Plant hormones play key roles in defence against pathogen attack. Recent work has begun to extend this role to encompass not just the traditional disease/stress hormones, such as ethylene, but also growth‐promoting hormones. Strigolactones (SLs) are the most recently defined group of plant hormones with important roles in plant–microbe interactions, as well as aspects of plant growth and development, although the knowledge of their role in plant–pathogen interactions is extremely limited. The oomycete Pythium irregulare is a poorly controlled pathogen of many crops. Previous work has indicated an important role for ethylene in defence against this oomycete. We examined the role of ethylene and SLs in response to this pathogen in pea (Pisum sativum L.) at the molecular and whole‐plant levels using a set of well‐characterized hormone mutants, including an ethylene‐insensitive ein2 mutant and SL‐deficient and insensitive mutants. We identified a key role for ethylene signalling in specific cell types that reduces pathogen invasion, extending the work carried out in other species. However, we found no evidence that SL biosynthesis or response influences the interaction of pea with P. irregulare or that synthetic SL influences the growth or hyphal branching of the oomycete in vitro. Future work should seek to extend our understanding of the role of SLs in other plant interactions, including with other fungal, bacterial and viral pathogens, nematodes and insect pests.
Ehlers Danlos syndrome and gastrointestinal manifestations: a 20‐year experience at Mayo Clinic
Authors: Nelson A. D. , Mouchli M. A. , Valentin N. , Deyle D. , Pichurin P. , Acosta A. , Camilleri M.
Published: September 2015

Background

Gastrointestinal (GI) manifestations are found in Ehlers Danlos syndrome (EDS) hypermobility subtype (HM). We aimed to assess associations between EDS HM and other EDS subtypes with GI manifestations.

Methods

We reviewed medical records of EDS patients evaluated at Mayo Clinic's Medical Genetics Clinic 1994–2013. We extracted information regarding EDS subtypes, GI manifestations, and treatments.

Key Results

We identified 687 patients; 378 (56%) had associated GI manifestations (female 86.8%, diagnosis mean age 29.6 years). Of the patients identified, 58.9% (43/73) had EDS classic, 57.5% (271/471) EDS HM, 47.3% (27/57) EDS vascular subtypes. In addition, 86 patients had EDS that could not be classified in any of those three subtypes. Commonest GI symptoms were: abdominal pain (56.1%), nausea (42.3%), constipation (38.6%), heartburn (37.6%), and irritable bowel syndrome‐like symptoms (27.5%). Many GI symptoms were commoner in EDS HM than the other subtypes together. Among 37.8% of the 378 patients who underwent esophagogastroduodenoscopy, the commonest abnormalities were gastritis, hiatal hernia and reflux esophagitis. Abnormal gastric emptying was observed in 22.3% (17/76): 11.8% delayed and 10.5% accelerated. Colonic transit was abnormal in 28.3% (13/46): 19.6% delayed and 8.7% accelerated. Rectal evacuation disorder was confirmed in 18/30 patients who underwent anorectal manometry. Angiography showed aneurysms in abdominal vessels in EDS vascular type. Proton pump inhibitors (38%) and drugs for constipation (23%) were the most commonly used medications. A minority underwent colectomy (2.9%) or small bowel surgery (4%).

Conclusions & Inferences

EDS HM and other subtypes should be considered in patients with chronic functional GI symptoms and abdominal vascular lesions.
Triviality For Restrictor Conditionals
Authors: Charlow Nate
Published: September 2015

I present two Triviality results for Kratzer's standard “restrictor” analysis of indicative conditionals (while also clarifying the sense in which Kratzer's semantics might avoid such results). I both refine and undermine the common claim that problems of Triviality do not arise for Kratzer conditionals since they are not strictly conditionals at all.
The carotenoid cleavage dioxygenase CCD2 catalysing the synthesis of crocetin in spring crocuses and saffron is a plastidial enzyme
Authors: Ahrazem Oussama , Rubio‐Moraga Angela , Berman Judit , Capell Teresa , Christou Paul , Zhu Changfu , Gómez‐Gómez Lourdes
Published: September 2015

The apocarotenoid crocetin and its glycosylated derivatives, crocins, confer the red colour to saffron. Crocetin biosynthesis in saffron is catalysed by the carotenoid cleavage dioxygenase CCD2 (AIG94929). No homologues have been identified in other plant species due to the very limited presence of crocetin and its derivatives in the plant kingdom.

Spring Crocus species with yellow flowers accumulate crocins in the stigma and tepals. Four carotenoid CCDs, namely CaCCD1, CaCCD2 and CaCCD4a/b and CaCCD4c were first cloned and characterized.

CaCCD2 was localized in plastids, and a longer CCD2 version, CsCCD2L, was also localized in this compartment. The activity of CaCCD2 was assessed in Escherichia coli and in a stable rice gene function characterization system, demonstrating the production of crocetin in both systems. The expression of all isolated CCDs was evaluated in stigma and tepals at three key developmental stages in relation with apocarotenoid accumulation. CaCCD2 expression parallels crocin accumulation, but C14 apocarotenoids most likely are associated to the CaCCD1 activity in Crocus ancyrensis flowers.

The specific CCD2 localization and its membrane interaction will contribute to the development of a better understanding of the mechanism of crocetin biosynthesis and regulation in the chromoplast.
An Empirical Analysis of the Municipal Bond Market in Italy: Sovereign Risk and Sub‐Sovereign Levels of Government
Authors: Pinna Massimo
Published: September 2015

This study empirically investigates the factors affecting municipal bond yields in the Italian market, a market characterized by several tiers of sub‐sovereign issuers and by a non‐negligible sovereign risk. Economic, financial and fiscal data from regions, provinces and cities are used to assess how a municipality's credit quality affects its borrowing costs. The results suggest that investors apply different pricing schemes to different types of local governments. Whereas the default risk of regions matters, the creditworthiness of cities and provinces is irrelevant, with sovereign risk and interest rate risk alone able to largely explain the variance in yields.
Ready to Bargain: The Effect of Fiscal Stress on Supermajority Requirements to Raise Taxes
Authors: Seljan Ellen C.
Published: September 2015

This paper analyzes the interaction effect between fiscal stress and supermajority requirements to raise taxes. I hypothesize that fiscal stress nullifies the effects of supermajority requirements, making states with and without supermajority requirements equally likely to raise taxes during these periods. This conclusion was drawn from a theory on how fiscal stress affects the legislative bargaining environment. The hypothesis was tested using data from 49 states from 1980 to 2010, and the results confirmed expectations. This research contributes to a growing literature that suggests that the effect of institutions cannot be analyzed without considering the context of the political environment.
Leviathan or Flypaper: Examining the Fungibility of Earmarked Local Sales Taxes for Transportation
Authors: Afonso Whitney B.
Published: September 2015

Analyses of the relationships of earmarked finances on their respective programs and total expenditures have produced conflicting views of how governments spend earmarked revenue. Some show that earmarks are used in addition to existing finances to increase the total level of funding for recipient programs while others show that earmarks are fungible and program spending is unchanged. An analysis on local option sales taxes earmarked for transportation (LOST‐Ts) suggests that transportation spending is increased by more than LOST‐T revenue. For every dollar generated by LOST‐Ts, transportation spending increases by an estimated $1.76 and nontransportation spending decreases by an estimated $0.73.
Injuries in male professional football: A prospective comparison between individual and team‐based exposure registration
Authors: Kristenson K. , Bjørneboe J. , Waldén M. , Andersen T. E. , Ekstrand J. , Hägglund M.
Published: September 2015

Methodological considerations of football injury epidemiology have only scarcely been described. The aim of this study was to evaluate the inter‐rater agreement in injury capture rate and injury categorization for data registered in two different prospective injury surveillance audits studying the same two Norwegian male professional football clubs for two consecutive seasons, 2008–2009. One audit used team‐based exposure (TBE) recording and the other individual‐based exposure (IBE). The number of injuries recorded and corresponding injury rates (injuries/1000 h exposure) were compared between audits. Cohen's kappa and prevalence‐adjusted bias‐adjusted kappa (PABAK) coefficients were calculated for injury variables. Of 323 injuries included, the IBE audit captured 318 (overall capture rate 98.5%, training 98.9%, match 97.8%) and the TBE audit 303 injuries (overall capture rate 93.8%, training 91.4%, match 97.1%). Agreement analysis showed kappa and PABAK coefficients regarded as almost perfect (> 0.81) for 8 of 9 injury variables, and substantial (ƙ 0.75) for the variable injury severity. In conclusion, the capture rate for training injuries was slightly higher with IBE recording, and inter‐agreement in injury categorization was very high.
Effect of patellar strap and sports tape on pain in patellar tendinopathy: A randomized controlled trial
Authors: de Vries A. , Zwerver J. , Diercks R. , Tak I. , van Berkel S. , van Cingel R. , van der Worp H. , van den Akker‐Scheek I.
Published: September 2015

Numerous athletes with patellar tendinopathy (PT) use a patellar strap or sports tape during sports. This study's aim was to investigate the short‐term effect of these orthoses on patellar tendon pain. Participants performed the single‐leg decline squat, vertical jump test, and triple‐hop test under four different conditions (patellar strap, sports tape, placebo, and control). Subsequently, participants practiced sports as usual for 2 weeks; during 1 week, they were assigned to one of the four conditions. Pain was measured with the visual analog scale (VAS). In total, 97 athletes with PT [61% male, age 27.0 (SD8.1), VISA‐P 58.5 (SD12.7)] were analyzed. On the single‐leg decline squat, the VAS pain score reduced significantly in the patellar strap (14 mm, P = 0.04) and the sports tape condition (13 mm, P = 0.04), compared with control, but not placebo. A significant decrease in VAS pain during sports was found in the sports tape (7 mm, P = 0.04) and placebo group (6 mm, P = 0.04). The VAS pain score two hours after sports decreased significantly in the patellar strap, sports tape and placebo group (8‐mm, P < 0.001, 10 mm, P = 0.001 and 7 mm, P = 0.03, respectively). This study's findings indicate that an orthosis (including placebo tape) during sports can reduce pain in PT patients in the short term.
Automatic conversion of IFC datasets to geometrically and semantically correct CityGML LOD3 buildings
Authors: Donkers Sjors , Ledoux Hugo , Zhao Junqiao , Stoter Jantien
Published: September 2015

Although the international standard CityGML has five levels of detail (LODs), the vast majority of available models are the coarse ones (up to LOD2, i.e. block‐shaped buildings with roofs). LOD3 and LOD4 models, which contain architectural details such as balconies, windows and rooms, rarely exist because, unlike coarser LODs, their construction requires several datasets that must be acquired with different technologies, and often extensive manual work is needed. In this article we investigate an alternative to obtaining CityGML LOD3 models: the automatic conversion from already existing architectural models (stored in the IFC format). Existing conversion algorithms mostly focus on the semantic mappings and convert all the geometries, which yields CityGML models having poor usability in practice (spatial analysis, for instance, is not possible). We present a conversion algorithm that accurately applies the correct semantics from IFC models and that constructs valid CityGML LOD3 buildings by performing a series of geometric operations in 3D. We have implemented our algorithm and we demonstrate its effectiveness with several real‐world datasets. We also propose specific improvements to both standards to foster their integration in the future.
Locking Compression Plate Fixation of Equine Tarsal Subluxations
Authors: Keller Sara A. , Fürst Anton E. , Kircher Patrick , Ringer Simone , Kuemmerle Jan M.
Published: September 2015

Objective

To report the clinical findings and outcome of locking compression plate (LCP) fixation of tarsal subluxations in horses.

Study Design

Retrospective case series.

Animals

Horses (n = 3) and ponies (2).

Methods

All horses and ponies diagnosed with tarsal subluxations and treated with LCP fixation at our institution between 2011 and 2013 were included. Data were collected from the medical records, including all radiographic and computed tomographic (CT) studies and reviewed. Long‐term (>1 year) followup examination, including owner interview, and clinical and radiographic examinations were performed for all cases.

Results

Subluxations of the tarsometatarsal (n = 3) and proximal intertarsal (2) joints were diagnosed. CT examinations revealed complex joint injuries, including a combination of avulsion fragments and compression injuries at the opposite side of the joint from the subluxation. The LCP was applied to the plantarolateral or medial side of the limb. All horses survived and were discharged from the hospital. Short‐term complications included mild laminitis (n = 1) and peroneus tertius rupture (2). At long‐term followup, both ponies were sound and used as intended, whereas all horses showed mild to moderate lameness at the trot and were used for pleasure riding only. Radiographic signs of osteoarthritis developed in the joint affected by subluxation in all cases and in adjacent small tarsal joints in 2 horses and 1 pony.

Conclusion

LCP fixation of tarsal subluxations resulted in stable fixation and allowed adequate healing. Subluxation of the small tarsal joints represents a complex injury and osteoarthritis commonly develops in the affected and adjacent joints.
Sleep spindles and human cortical nociception: a surface and intracerebral electrophysiological study
Authors: Claude Léa , Chouchou Florian , Prados Germán , Castro Maïté , De Blay Barbara , Perchet Caroline , García‐Larrea Luis , Mazza Stéphanie , Bastuji Hélène
Published: September 2015

Key points

Sleep spindle are usually considered to play a major role in inhibiting sensory inputs.

Using nociceptive stimuli in humans, we tested the effect of spindles on behavioural, autonomic and cortical responses in two experiments using surface and intracerebral electroencephalographic recordings.

We found that sleep spindles do not prevent arousal reactions to nociceptive stimuli and that autonomic reactivity to nociceptive inputs is not modulated by spindle activity.

Moreover, neither the surface sensory, nor the insular evoked responses were modulated by the spindle, as detected at the surface or within the thalamus.

The present study comprises the first investigation of the effect of spindles on nociceptive information processing and the results obtained challenge the classical inhibitory effect of spindles.

Responsiveness to environmental stimuli declines during sleep, and sleep spindles are often considered to play a major role in inhibiting sensory inputs. In the present study, we tested the effect of spindles on behavioural, autonomic and cortical responses to pain, in two experiments assessing surface and intracerebral responses to thermo‐nociceptive laser stimuli during the all‐night N2 sleep stage. The percentage of arousals remained unchanged as a result of the presence of spindles. Neither cortical nociceptive responses, nor autonomic cardiovascular reactivity were depressed when elicited within a spindle. These results could be replicated in human intracerebral recordings, where sleep spindle activity in the posterior thalamus failed to depress the thalamocortical nociceptive transmission, as measured by sensory responses within the posterior insula. Hence, the assumed inhibitory effect of spindles on sensory inputs may not apply to the nociceptive system, possibly as a result of the specificity of spinothalamic pathways and the crucial role of nociceptive information for homeostasis. Intriguingly, a late scalp response commonly considered to reflect high‐order stimulus processing (the ‘P3’ potential) was significantly enhanced during spindling, suggesting a possible spindle‐driven facilitation, rather than attenuation, of cortical nociception.
Mitochondria and carbon monoxide: cytoprotection and control of cell metabolism – a role for Ca2+?
Authors: R. Oliveira Sara , Queiroga Cláudia S. F. , Vieira Helena L. A.
Published: September 2015

Carbon monoxide (CO) is an endogenously produced gasotransmitter with important biological functions: anti‐inflammation, anti‐apoptosis, vasomodulation and cell metabolism modulation. The most recognized cellular target for CO is the mitochondria. Physiological concentrations of CO generate mitochondrial reactive oxygen species (ROS), which are signalling molecules for CO‐induced pathways. Indeed, small amounts of ROS promote cytoprotection by a preconditioning effect. Furthermore, CO prevents cell death by limiting mitochondrial membrane permeabilization, which inhibits the release of pro‐apoptotic factors into the cytosol; both events are ROS dependent. CO also increases the ability of mitochondria to take up Ca2+. Mitochondrial metabolism is modulated by CO, namely by increasing TCA cycle rate, oxidative phosphorylation and mitochondrial biogenesis, which, in turn, increases ATP production. CO's modulation of metabolism might be important for cellular response to diseases, namely cancer and ischaemic diseases. Finally, another cytoprotective role of CO involves the control of Ca2+ channels. By limiting the activity of T‐type and L‐type Ca2+ channels, CO prevents excitotoxicity‐induced cell death and modulates cell proliferation. Several questions concerning Ca2+ signalling, mitochondria and CO can be asked, for instance whether CO modulation of cell metabolism would be dependent on the mitochondrial Ca2+ uptake capacity, since small amounts of Ca2+ can increase mitochondrial metabolism. Whether CO controls Ca2+ communication between mitochondria and endoplasmic reticulum is another open field of research. In summary, CO emerges as a key gasotransmitter in the control of several cellular functions of mitochondria: metabolism, cell death and Ca2+ signalling.

image
Shear stress induces a longitudinal Ca2+ wave via autocrine activation of P2Y1 purinergic signalling in rat atrial myocytes
Authors: Kim Joon‐Chul , Woo Sun‐Hee
Published: September 2015

Key points

Cardiac myocytes are subjected to fluid shear stress during the cardiac cycle and haemodynamic disturbance.

A longitudinally propagating, regenerative Ca2+ wave is initiated in atrial myocytes under shear stress.

Here we determine the cellular mechanism for this shear‐induced Ca2+ wave using two‐dimensional confocal Ca2+ imaging combined with pressurized fluid flow.

Our data suggest that shear stress triggers the Ca2+ wave through ryanodine receptors via P2Y1 purinoceptor–phospholipase C‐type 2 inositol 1,4,5‐trisphosphate receptor signal transduction in atrial myocytes, and that this mechanotransduction is activated by gap junction hemichannel‐mediated ATP release.

Shear‐specific mechanotransduction and the subsequent regenerative Ca2+ wave may be one way for atrial myocytes to assess mechanical stimuli directly and alter their Ca2+ signalling accordingly.

Atrial myocytes are exposed to shear stress during the cardiac cycle and haemodynamic disturbance. In response, they generate a longitudinally propagating global Ca2+ wave. Here, we investigated the cellular mechanisms underlying the shear stress‐mediated Ca2+ wave, using two‐dimensional confocal Ca2+ imaging combined with a pressurized microflow system in single rat atrial myocytes. Shear stress of ∼16 dyn cm−2 for 8 s induced ∼1.2 aperiodic longitudinal Ca2+ waves (∼79 μm s−1) with a delay of 0.2−3 s. Pharmacological blockade of ryanodine receptors (RyRs) or inositol 1,4,5‐trisphosphate receptors (IP3Rs) abolished shear stress‐induced Ca2+ wave generation. Furthermore, in atrial myocytes from type 2 IP3R (IP3R2) knock‐out mice, shear stress failed to induce longitudinal Ca2+ waves. The phospholipase C (PLC) inhibitor U73122, but not its inactive analogue U73343, abolished the shear‐induced longitudinal Ca2+ wave. However, pretreating atrial cells with blockers for stretch‐activated channels, Na+−Ca2+ exchanger, transient receptor potential melastatin subfamily 4, or nicotinamide adenine dinucleotide phosphate oxidase did not suppress wave generation under shear stress. The P2 purinoceptor inhibitor suramin, and the potent P2Y1 receptor antagonist MRS 2179, both suppressed the Ca2+ wave, whereas the P2X receptor antagonist, iso‐PPADS, did not alter it. Suppression of gap junction hemichannels permeable to ATP or extracellular application of ATP‐metabolizing apyrase inhibited the wave. Removal of external Ca2+ to enhance hemichannel opening facilitated the wave generation. Our data suggest that longitudinally propagating, regenerative Ca2+ release through RyRs is triggered by P2Y1–PLC–IP3R2 signalling that is activated by gap junction hemichannel‐mediated ATP release in atrial myocytes under shear stress.
Commedia dell'Arte as a Metaphor for the Art of Teaching
Authors: Morse Robert A.
Published: September 2015

With my background in acting, stagecraft, and some knowledge of theatre history from high school and college, it was natural for me as a novice teacher to think about theatre as a metaphor for teaching. In this article I will discuss how over my career I conceived of teaching in terms of theatrical metaphors, and make a comparison with a common metaphor of teaching as coaching.
Teaching the Delightful Laws of Physics in a Survey Course
Authors: Hewitt Paul G.
Published: September 2015

How physics can be made interesting is a question that needs no answer. That's because physics is interesting! It's a field of study jam-packed with fascination and wonder. The general public has an enormous thirst for physics knowledge, as indicated by the great numbers who purchase science magazines and books and watch NOVA and other science specials. A related question, how to make a physics course interesting, is something that we can answer. All we have to do is present physics at a proper pace in the language of the learner. My adage has always been that if the first course in physics is delightful, the rigor of a follow-up course will be welcomed.
Availability of high school physics and socioeconomics
Authors: White Susan C.
Published: September 2015

We noted last month that Hispanic and African-American seniors were less likely to have taken a high school physics course than their peers, and we suggested that socioeconomic status (SES) played a role in the lower participation. Often the proportion of students receiving free and reduced-price meals (FRMs) is used as a measure of SES in secondary education; however, we found that this measure did not differentiate very well between schools at the upper end of the socioeconomic spectrum. To help us understand both ends of the SES scale, we ask principals and teachers to assess the economic situation of their students relative to others in the area. We have found that this tracks well with FRMs on the low end and helps us distinguish between schools on the upper end. This figure displays the number of seniors in public high schools attending a school where physics is never offered by SES. While the number of seniors is roughly one-third in each SES group, over half of the seniors attending a school where physics is not offered are at “worse off” schools. Next month we will look more closely at physics by SES.
Workshop Physics and Related Curricula: A 25-Year History of Collaborative Learning Enhanced by Computer Tools for Observation and Analysis
Authors: Laws Priscilla W. , Willis Maxine C. , Sokoloff David R.
Published: September 2015

This article describes the 25-year history of development of the activity-based Workshop Physics (WP) at Dickinson College, its adaptation for use at Gettysburg Area High School, and its synergistic influence on curricular materials developed at the University of Oregon and Tufts University and vice versa. WP and these related curricula: 1) are based on Physics Education Research (PER) findings and are PER-validated; 2) feature active, collaborative learning; and 3) use computer-based tools that enable students to learn by making predictions and then collecting, displaying, and analyzing data from their experiments.
Teaching Physics with Music
Authors: Ramsey Gordon P.
Published: September 2015

The uniting of two seemingly disparate subjects in the classroom provides an interesting motivation for learning. Students are interested in how these subjects can possibly be integrated into related ideas. Such is the mixture of physics and music. Both are based upon mathematics, which becomes the interlocking theme. The connecting physical properties of sound and music are waves and harmonics. The introduction of instruments, including the voice, to the musical discussion allows the introduction of more advanced physical concepts such as energy, force, pressure, fluid dynamics, and properties of materials. Suggestions on how to teach physics concepts in the context of music at many levels are presented in this paper.
The Physlet Approach to Simulation Design
Authors: Christian Wolfgang , Belloni Mario , Esquembre Francisco , Mason Bruce A. , Barbato Lyle , Riggsbee Matt
Published: September 2015

Over the past two years, the AAPT/ComPADRE staff and the Open Source Physics group have published the second edition of Physlet Physics and Physlet Quantum Physics, delivered as interactive web pages on AAPT/ComPADRE and as free eBooks available through iTunes and Google Play. These two websites, and their associated books, add over 1000 interactive exercises for the teaching of introductory physics, introductory and intermediate modern physics, and quantum mechanics to AAPT/ComPADRE.
Color Addition and Subtraction Apps
Authors: Ruiz Frances , Ruiz Michael J.
Published: September 2015

Color addition and subtraction apps in HTML5 have been developed for students as an online hands-on experience so that they can more easily master principles introduced through traditional classroom demonstrations. The evolution of the additive RGB color model is traced through the early IBM color adapters so that students can proceed step by step in understanding mathematical representations of RGB color. Finally, color addition and subtraction are presented for the X11 colors from web design to illustrate yet another real-life application of color mixing.
Understanding the Association of Human Rhinovirus with Asthma
Authors: Stone Cosby A. , Miller E. Kathryn
Published: September 2015

Human rhinoviruses are ubiquitous seasonal pathogens. They have known associations with first onset of wheezing illnesses in children and with asthma exacerbations in patients of all ages. It is not yet certain whether human rhinoviruses play a direct role in the pathogenesis of asthma by activating deleterious inflammatory responses or if they only serve as a catalyst to accelerate the disease in genetically predisposed individuals. There have been previously demonstrated reductions in the development of the asthmatic phenotype with passive immunization against respiratory syncytial virus; however, in the case of rhinovirus, there are barriers to effective vaccine development, such as the lack of a common antigenic target due to alterations of surface markers among subtypes. It remains to be determined whether certain subtypes of human rhinovirus are more asthmagenic and therefore worthy of greater attention as vaccine candidates, but several studies have suggested that RV-C and certain RV-A strains may be more strongly linked with asthma.
Mapping Type IV Secretion Signals on the Primase Encoded by the Broad-Host-Range Plasmid R1162 (RSF1010)
Authors: Meyer Richard
Published: September 2015

The plasmid R1162 (RSF1010) encodes a primase essential for its replication. This primase makes up the C-terminal part of MobA, a multifunctional protein with the relaxase as a separate N-terminal domain. The primase is also translated separately as the protein RepB′. Here, we map two signals for type IV secretion onto the recently solved structure of RepB′. One signal is located internally within RepB′ and consists of a long α-helix and an adjacent disordered region rich in arginines. The second signal is made up of the same α-helix and a second, arginine-rich region at the C-terminal end of the protein. Successive arginine-to-alanine substitutions revealed that either signal can be utilized by the type IV secretion complex of the plasmid R751. The internal signal also enables conjugal transfer when linked to the relaxase part of MobA. Both signals are similar to those previously identified for type IV secretion substrates in the Vir system of
Agrobacterium tumefaciens
. Moreover, the C-terminal arginine-rich segment of RepB′ has been shown to be secreted by Vir. However, with R751, the signals require MobB, an R1162-encoded accessory protein active in conjugal transfer. The results of two-hybrid assays revealed that MobB interacts, via its membrane-associated domain, with the R751 plasmid coupling protein TraG. In addition, MobB interacts with a region of MobA just outside the RepB′ domain. Therefore, MobB is likely an adaptor that is essential for recognition of the primase-associated signals by the R751 secretion machinery.

IMPORTANCE
For most plasmids, type IV secretion is an intrinsic part of the mechanism for conjugal transfer. Protein relaxases, bound to the 5′ end of the transferring strand, are mobilized into recipient cells by the type IV pathway. In this work, we identify and characterize two signals for secretion in the primase domain of MobA, the relaxase of the IncQ plasmid R1162 (RSF1010). We also show that the adaptor protein MobB is required for engagement of these signals with the R751 coupling protein TraG. These results clarify the location and properties of secretion signals active during the conjugal transfer of plasmid DNA.
First Report of Wohlfahrtiimonaschitiniclastica Isolation from a Patient with Cellulitis in the United States
Authors: de Dios Angel , Jacob Seby , Tayal Amit , Fisher Mark A. , Dingle Tanis C. , Hamula Camille L.
Published: September 2015

We report the first documented isolation of
Wohlfahrtiimonas
chitiniclastica
from a human in the United States. Initially misidentified as
Acinetobacter lwoffii
by Vitek-2, the isolate was subsequently identified as
W
.
chitiniclastica
by matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) and 16S rRNA gene sequencing. While the clinical significance of the isolate in this case is unclear, it highlights the superior performance of MALDI-TOF MS for bacterial identification.
Metabolomic Profiling of Plasma from Patients with Tuberculosis by Use of Untargeted Mass Spectrometry Reveals Novel Biomarkers for Diagnosis
Authors: Lau Susanna K. P. , Lee Kim-Chung , Curreem Shirly O. T. , Chow Wang-Ngai , To Kelvin K. W. , Hung Ivan F. N. , Ho Deborah T. Y. , Sridhar Siddharth , Li Iris W. S. , Ding Vanessa S. Y. , Koo Eleanor W. F. , Wong Chi-Fong , Tam Sidney , Lam Ching-Wan , Yuen Kwok-Yung , Woo Patrick C. Y.
Published: September 2015

Although tuberculosis (TB) is a reemerging disease that affects people in developing countries and immunocompromised populations in developed countries, the current diagnostic methods are far from optimal. Metabolomics is increasingly being used for studies on infectious diseases. We performed metabolome profiling of plasma samples to identify potential biomarkers for diagnosing TB. We compared the plasma metabolome profiles of TB patients (
n
= 46) with those of community-acquired pneumonia (CAP) patients (
n
= 30) and controls without active infection (
n
= 30) using ultrahigh-performance liquid chromatography–electrospray ionization-quadrupole time of flight mass spectrometry (UHPLC-ESI-QTOFMS). Using multivariate and univariate analyses, four metabolites, 12R-hydroxy-5Z,8Z,10E,14Z-eicosatetraenoic acid [12(R)-HETE], ceramide (d18:1/16:0), cholesterol sulfate, and 4α-formyl-4β-methyl-5α-cholesta-8-en-3β-ol, were identified and found to have significantly higher levels in TB patients than those in CAP patients and controls. In a comparison of TB patients and controls, the four metabolites demonstrated area under the receiver operating characteristic curve (AUC) values of 0.914, 0.912, 0.905, and 0.856, sensitivities of 84.8%, 84.8%, 87.0%, and 89.1%, specificities of 90.0%, 86.7%, 86.7%, and 80.0%, and fold changes of 4.19, 26.15, 6.09, and 1.83, respectively. In a comparison of TB and CAP patients, the four metabolites demonstrated AUC values of 0.793, 0.717, 0.802, and 0.894, sensitivities of 89.1%, 71.7%, 80.4%, and 84.8%, specificities of 63.3%, 66.7%, 70.0%, and 83.3%, and fold changes of 4.69, 3.82, 3.75, and 2.16, respectively. 4α-Formyl-4β-methyl-5α-cholesta-8-en-3β-ol combined with 12(R)-HETE or cholesterol sulfate offered ≥70% sensitivity and ≥90% specificity for differentiating TB patients from controls or CAP patients. These novel plasma biomarkers, especially 12(R)-HETE and 4α-formyl-4β-methyl-5α-cholesta-8-en-3β-ol, alone or in combination, are potentially useful for rapid and noninvasive diagnosis of TB. The present findings may offer insights into the pathogenesis and host response in TB.
One-Step Identification of Five Prominent Chicken Salmonella Serovars and Biotypes
Authors: Zhu Chunhong , Yue Min , Rankin Shelley , Weill François-Xavier , Frey Joachim , Schifferli Dieter M.
Published: September 2015

Based on bacterial genomic data, we developed a one-step multiplex PCR assay to identify
Salmonella
and simultaneously differentiate the two invasive avian-adapted
S. enterica
serovar Gallinarum biotypes Gallinarum and Pullorum, and the most frequent, specific, and asymptomatic colonizers of chickens, serovars Enteritidis, Heidelberg, and Kentucky.
Characterization of a Novel Hepadnavirus in the White Sucker (Catostomus commersonii) from the Great Lakes Region of the United States
Authors: Hahn Cassidy M. , Iwanowicz Luke R. , Cornman Robert S. , Conway Carla M. , Winton James R. , Blazer Vicki S.
Published: September 2015

The white sucker
Catostomus commersonii
is a freshwater teleost often utilized as a resident sentinel. Here, we sequenced the full genome of a hepatitis B-like virus that infects white suckers from the Great Lakes Region of the United States. Dideoxy sequencing confirmed that the white sucker hepatitis B virus (WSHBV) has a circular genome (3,542 bp) with the prototypical codon organization of hepadnaviruses. Electron microscopy demonstrated that complete virions of approximately 40 nm were present in the plasma of infected fish. Compared to avi- and orthohepadnaviruses, sequence conservation of the core, polymerase, and surface proteins was low and ranged from 16 to 27% at the amino acid level. An X protein homologue common to the orthohepadnaviruses was not present. The WSHBV genome included an atypical, presumptively noncoding region absent in previously described hepadnaviruses. Phylogenetic analyses confirmed WSHBV as distinct from previously documented hepadnaviruses. The level of divergence in protein sequences between WSHBV and other hepadnaviruses and the identification of an HBV-like sequence in an African cichlid provide evidence that a novel genus of the family
Hepadnaviridae
may need to be established that includes these hepatitis B-like viruses in fishes. Viral transcription was observed in 9.5% (16 of 169) of white suckers evaluated. The prevalence of hepatic tumors in these fish was 4.9%, and only 2.4% of fish were positive for both virus and hepatic tumors. These results are not sufficient to draw inferences regarding the association of WSHBV and carcinogenesis in white sucker.

IMPORTANCE
We report the first full-length genome of a hepadnavirus from fishes. Phylogenetic analysis of this genome indicates divergence from genomes of previously described hepadnaviruses from mammalian and avian hosts and supports the creation of a novel genus. The discovery of this novel virus may better our understanding of the evolutionary history of hepatitis B-like viruses of other hosts. In fishes, knowledge of this virus may provide insight regarding possible risk factors associated with hepatic neoplasia in the white sucker. This may also offer another model system for mechanistic research.
Dengue Virus NS1 Protein Modulates Cellular Energy Metabolism by Increasing Glyceraldehyde-3-Phosphate Dehydrogenase Activity
Authors: Allonso Diego , Andrade Iamara S. , Conde Jonas N. , Coelho Diego R. , Rocha Daniele C. P. , da Silva Manuela L. , Ventura Gustavo T. , Silva Emiliana M. , Mohana-Borges Ronaldo
Published: September 2015

Dengue is one of the main public health concerns worldwide. Recent estimates indicate that over 390 million people are infected annually with the dengue virus (DENV), resulting in thousands of deaths. Among the DENV nonstructural proteins, the NS1 protein is the only one whose function during replication is still unknown. NS1 is a 46- to 55-kDa glycoprotein commonly found as both a membrane-associated homodimer and a soluble hexameric barrel-shaped lipoprotein. Despite its role in the pathogenic process, NS1 is essential for proper RNA accumulation and virus production. In the present study, we identified that glyceraldehyde-3-phosphate dehydrogenase (GAPDH) interacts with intracellular NS1. Molecular docking revealed that this interaction occurs through the hydrophobic protrusion of NS1 and the hydrophobic residues located at the opposite side of the catalytic site. Moreover, addition of purified recombinant NS1 enhanced the glycolytic activity of GAPDH
in vitro
. Interestingly, we observed that DENV infection promoted the relocalization of GAPDH to the perinuclear region, where NS1 is commonly found. Both DENV infection and expression of NS1 itself resulted in increased GAPDH activity. Our findings indicate that the NS1 protein acts to increase glycolytic flux and, consequently, energy production, which is consistent with the recent finding that DENV induces and requires glycolysis for proper replication. This is the first report to propose that NS1 is an important modulator of cellular energy metabolism. The data presented here provide new insights that may be useful for further drug design and the development of alternative antiviral therapies against DENV.

IMPORTANCE
Dengue represents a serious public health problem worldwide and is caused by infection with dengue virus (DENV). Estimates indicate that half of the global population is at risk of infection, with almost 400 million cases occurring per year. The NS1 glycoprotein is found in both the intracellular and the extracellular milieus. Despite the fact that NS1 has been commonly associated with DENV pathogenesis, it plays a pivotal but unknown role in the replication process. In an effort to understand the role of intracellular NS1, we demonstrate that glyceraldehyde-3-phosphate dehydrogenase (GAPDH) interacts with NS1. Our results indicate that NS1 increases the glycolytic activity of GAPDH
in vitro
. Interestingly, the GAPDH activity was increased during DENV infection, and NS1 expression alone was sufficient to enhance intracellular GAPDH activity in BHK-21 cells. Overall, our findings suggest that NS1 is an important modulator of cellular energy metabolism by increasing glycolytic flux.
CD8lowCD100−T Cells Identify a Novel CD8 T Cell Subset Associated with Viral Control during Human Hantaan Virus Infection
Authors: Liu Bei , Ma Ying , Zhang Yusi , Zhang Chunmei , Yi Jing , Zhuang Ran , Yu Haitao , Yang Angang , Zhang Yun , Jin Boquan
Published: September 2015

Hantaan virus (HTNV) infection can cause a severe lethal hemorrhagic fever with renal syndrome (HFRS) in humans. CD8+T cells play a critical role in combating HTNV infections. However, the contributions of different CD8+T cell subsets to the immune response against viral infection are poorly understood. Here, we identified a novel subset of CD8+T cells characterized by the CD8lowCD100−phenotype in HFRS patients. The CD8lowCD100−subset accounted for a median of 14.3% of the total CD8+T cells in early phase of HFRS, and this percentage subsequently declined in the late phase of infection, whereas this subset was absent in healthy controls. Furthermore, the CD8lowCD100−cells were associated with high activation and expressed high levels of cytolytic effector molecules and exhibited a distinct expression profile of effector CD8+T cells (CCR7+/−CD45RA−CD127highCD27intCD28lowCD62L−). When stimulated with specific HTNV nucleocapsid protein-derived peptide pools, most responding CD8+cells (gamma interferon [IFN-γ] positive and/or tumor necrosis factor alpha [TNF-α] positive) were CD8lowCD100−cells. The frequency of CD8lowCD100−cells among HTNV-specific CD8+T cells was higher in milder cases than in more severe cases. Importantly, the proportion of the CD8lowCD100−subset among CD8+T cells in early phase of HFRS was negatively correlated with the HTNV viral load, suggesting that CD8lowCD100−cells may be associated with viral clearance. The contraction of the CD8lowCD100−subset in late phase of infection may be related to the consistently high expression levels of PD-1. These results may provide new insights into our understanding of CD8+T cell-mediated protective immunity as well as immune homeostasis after HTNV infection in humans.

IMPORTANCECD8+T cells play important roles in the antiviral immune response. We found that the proportion of CD8lowCD100−cells among CD8+T cells from HFRS patients was negatively correlated with the HTNV viral load, and the frequency of CD8lowCD100−cells among virus-specific CD8+T cells was higher in milder HFRS cases than in more severe cases. These results imply a beneficial role for the CD8lowCD100−cell subset in viral control during human HTNV infection.
Differential Reovirus-Specific and Herpesvirus-Specific Activator Protein 1 Activation of Secretogranin II Leads to Altered Virus Secretion
Authors: Berard Alicia R. , Severini Alberto , Coombs Kevin M.
Published: September 2015

Viruses utilize host cell machinery for propagation and manage to evade cellular host defense mechanisms in the process. Much remains unknown regarding how the host responds to viral infection. We recently performed global proteomic screens of mammalian reovirus TIL- and T3D-infected and herpesvirus (herpes simplex virus 1 [HSV-1])-infected HEK293 cells. The nonenveloped RNA reoviruses caused an upregulation, whereas the enveloped DNA HSV-1 caused a downregulation, of cellular secretogranin II (SCG2). SCG2, a member of the granin family that functions in hormonal peptide sorting into secretory vesicles, has not been linked to virus infections previously. We confirmed SCG2 upregulation and found SCG2 phosphorylation by 18 h postinfection (hpi) in reovirus-infected cells. We also found a decrease in the amount of reovirus secretion from SCG2 knockdown cells. Similar analyses of cells infected with HSV-1 showed an increase in the amount of secreted virus. Analysis of the stress-activated protein kinase (SAPK)/Jun N-terminal protein kinase (JNK) pathway indicated that each virus activates different pathways leading to activator protein 1 (AP-1) activation, which is the known SCG2 transcription activator. We conclude from these experiments that the negative correlation between SCG2 quantity and virus secretion for both viruses indicates a virus-specific role for SCG2 during infection.

IMPORTANCEMammalian reoviruses affect the gastrointestinal system or cause respiratory infections in humans. Recent work has shown that all mammalian reovirus strains (most specifically T3D) may be useful oncolytic agents. The ubiquitous herpes simplex viruses cause common sores in mucosal areas of their host and have coevolved with hosts over many years. Both of these virus species are prototypical representatives of their viral families, and investigation of these viruses can lead to further knowledge of how they and the other more pathogenic members of their respective families interact with the host. Here we show that secretogranin II (SCG2), a protein not previously studied in the context of virus infections, alters virus output in a virus-specific manner and that the quantity of SCG2 is inversely related to amounts of infectious-virus secretion. Herpesviruses may target this protein to facilitate enhanced virus release from the host.
Differential Susceptibilities of Human Lung Primary Cells to H1N1 Influenza Viruses
Authors: Travanty Emily , Zhou Bin , Zhang Hongbo , Di Y. Peter , Alcorn John F. , Wentworth David E. , Mason Robert , Wang Jieru
Published: September 2015

Human alveolar epithelial cells (AECs) and alveolar macrophages (AMs) are the first lines of lung defense. Here, we report that AECs are the direct targets for H1N1 viruses that have circulated since the 2009 pandemic (H1N1pdm09). AMs are less susceptible to H1N1pdm09 virus, but they produce significantly more inflammatory cytokines than AECs from the same donor. AECs form an intact epithelial barrier that is destroyed by H1N1pdm09 infection. However, there is significant variation in the cellular permissiveness to H1N1pdm09 infection among different donors. AECs from obese donors appear to be more susceptible to H1N1pdm09 infection, whereas gender, smoking history, and age do not appear to affect AEC susceptibility. There is also a difference in response to different strains of H1N1pdm09 viruses. Compared to A/California04/09 (CA04), A/New York/1682/09 (NY1682) is more infectious and causes more epithelial barrier injury, although it stimulates less cytokine production. We further determined that a single amino acid residue substitution in NY1682 hemagglutinin is responsible for the difference in infectivity. In conclusion, this is the first study of host susceptibility of human lung primary cells and the integrity of the alveolar epithelial barrier to influenza. Further elucidation of the mechanism of increased susceptibility of AECs from obese subjects may facilitate the development of novel protection strategies against influenza virus infection.

IMPORTANCE
Disease susceptibility of influenza is determined by host and viral factors. Human alveolar epithelial cells (AECs) form the key line of lung defenses against pathogens. Using primary AECs from different donors, we provided cellular level evidence that obesity might be a risk factor for increased susceptibility to influenza. We also compared the infections of two closely related 2009 pandemic H1N1 strains in AECs from the same donor and identified a key viral factor that affected host susceptibility, the dominance of which may be correlated with disease epidemiology. In addition, primary human AECs can serve as a convenient and powerful model to investigate the mechanism of influenza-induced lung injury and determine the effect of genetic and epigenetic factors on host susceptibility to pandemic influenza virus infection.