bluuebunny/crossref_metadata_2025 huggingface dataset:
ZmSTK1 and ZmSTK2, encoding receptor‐like cytoplasmic kinase, are involved in maize pollen development with additive effect
Authors: Fan Mingxia , Zhang Chunyu , Shi Lei , Liu Chen , Ma Wenjuan , Chen Meiming , Liu Kuichen , Cai Fengchun , Wang Guohong , Wei Zhengyi , Jiang Min , Liu Zaochang , Javeed Ansar , Lin Feng
Published: January 2018

Pollen germination and pollen tube growth are important physiological processes of sexual reproduction of plants and also are involved in signal transduction. Our previous study reveals that ZmSTK1 and ZmSTK2 are two receptor‐like cytoplasmic kinases (RLCK) homologs in Zea mays as members of receptor‐like protein kinase (RLK) subfamily, sharing 86% identity at the amino acid level. Here, we report that ZmSTK1 and ZmSTK2, expressed at late stages of pollen development, regulate maize pollen development with additive effect. ZmSTK1 or ZmSTK2 mutation exhibited severe pollen transmission deficiency, which thus influenced pollen fertility. Moreover, the kinase domains of ZmSTKs were cross‐interacted with C‐terminus of enolases detected by co‐immunoprecipitation (Co‐IP) and yeast two‐hybrid system (Y2H), respectively. Further, the detective ZmSTK1 or ZmSTK2 was associated with decreased activity of enolases and also reduced downstream metabolite contents, which enolases are involved in glycolytic pathway, such as phosphoenolpyruvate (PEP), pyruvate, ADP/ATP, starch, glucose, sucrose and fructose. This study reveals that ZmSTK1 and ZmSTK2 regulate maize pollen development and indirectly participate in glycolytic pathway.
Biogenesis and regulatory hierarchy of phased small interfering RNAs in plants
Authors: Deng Pingchuan , Muhammad Sajid , Cao Min , Wu Liang
Published: January 2018

Several varieties of small RNAs including microRNAs (miRNAs) and small interfering RNAs (siRNAs) are generated in plants to regulate development, genome stability and response to adverse environments. Phased siRNA (phasiRNA) is a type of secondary siRNA that is processed from a miRNA‐mediated cleavage of RNA transcripts, increasing silencing efficiency or simultaneously suppressing multiple target genes. Trans‐acting siRNAs (ta‐siRNAs) are a particular class of phasiRNA produced from noncoding transcripts that silence targets in trans. It was originally thought that ‘one‐hit’ and ‘two‐hit’ models were essential for processing distinct TAS precursors; however, a single hit event was recently shown to be sufficient at triggering all types of ta‐siRNAs. This review discusses the findings about biogenesis, targeting modes and regulatory networks of plant ta‐siRNAs. We also summarize recent advances in the generation of other phasiRNAs and their possible biological benefits to plants.
For Those Who Care: The Effect of Public Service Motivation on Sector Selection
Authors: Holt Stephen B.
Published: January 2018

Public service motivation (PSM) theory suggests that the alignment of values may explain sorting into public service work. Evidence suggests that people with high PSM cluster in government and nonprofit organizations. However, reliance on cross‐sectional data leaves open the question of whether observed patterns are the result of public and nonprofit organizations attracting and selecting high‐PSM people or cultivating PSM through socialization within the sector. Using longitudinal data, this article analyzes the relationship between motivational bases, such as PSM, and sorting into the public, for‐profit, and nonprofit sectors. The results indicate that PSM‐related values, measured before labor market entry, predict the sector a person will select for employment. Moreover, the effect on sector selection does not operate through some commonly cited alternative predictor of sector employment, such as college completion. Rather, PSM predicts sorting into college majors in a manner consistent with sector sorting in the labor market.
Pneumonia as a cardiovascular disease
Authors: Restrepo Marcos I. , Reyes Luis F.
Published: January 2018

Community‐acquired pneumonia (CAP) is an important cause of death around the globe. Up to 30% of patients admitted to hospital for CAP develop cardiovascular complications (i.e. new/worsening heart failure, new/worsening arrhythmias, myocardial infarctions and/or strokes), acutely and up to 10 years thereafter. Cardiac complications result from complex interactions between preexisting conditions, relative ischaemia, upregulation of the sympathetic system, systemic inflammation and direct pathogen‐mediated damage to the cardiovascular system. The exact mechanisms underlying the direct host–pathogen interactions are of great interest to identify potential therapeutic and preventative targets for CAP. In this review, we summarize the epidemiological data, risk factors and the pathogen‐driven cardiovascular damage affecting patients with CAP.
Following the cold: geographical differentiation between interglacial refugia and speciation in the arcto‐alpine species complex Bombus monticola (Hymenoptera: Apidae)
Authors: MARTINET BAPTISTE , LECOCQ THOMAS , BRASERO NICOLAS , BIELLA PAOLO , URBANOVÁ KLÁRA , VALTEROVÁ IRENA , CORNALBA MAURIZIO , GJERSHAUG JAN OVE , MICHEZ DENIS , RASMONT PIERRE
Published: January 2018

Cold‐adapted species are expected to have reached their largest distribution range during a part of the Ice Ages whereas postglacial warming has led to their range contracting toward high‐latitude and high‐altitude areas. This has resulted in an extant allopatric distribution of populations and possibly to trait differentiations (selected or not) or even speciation. Assessing inter‐refugium differentiation or speciation remains challenging for such organisms because of sampling difficulties (several allopatric populations) and disagreements on species concept. In the present study, we assessed postglacial inter‐refugia differentiation and potential speciation among populations of one of the most common arcto‐alpine bumblebee species in European mountains, Bombus monticola Smith, 1849. Based on mitochondrial DNA/nuclear DNA markers and eco‐chemical traits, we performed integrative taxonomic analysis to evaluate alternative species delimitation hypotheses and to assess geographical differentiation between interglacial refugia and speciation in arcto‐alpine species. Our results show that trait differentiations occurred between most Southern European mountains (i.e. Alps, Balkan, Pyrenees, and Apennines) and Arctic regions. We suggest that the monticola complex actually includes three species: B. konradini stat.n. status distributed in Italy (Central Apennine mountains), B. monticola with five subspecies, including B. monticola mathildis ssp.n. distributed in the North Apennine mountains ; and B. lapponicus. Our results support the hypothesis that post‐Ice Age periods can lead to speciation in cold‐adapted species through distribution range contraction. We underline the importance of an integrative taxonomic approach for rigorous species delimitation, and for evolutionary study and conservation of taxonomically challenging taxa.
Molecular phylogenetics and piercer evolution in the bug‐killing flies (Diptera: Tachinidae: Phasiinae)
Authors: BLASCHKE JEREMY D. , STIREMAN JOHN O. , O'HARA JAMES E. , CERRETTI PIERFILIPPO , MOULTON JOHN K.
Published: January 2018

Phasiinae (Diptera: Tachinidae) are endoparasitoid flies that attack Heteroptera, including a multitude of agricultural pests. A phylogenetically informed classification of Phasiinae has eluded systematists for over a century, primarily because of the conflicting character states and confusing morphology of certain taxa that indicate potential placement within other subfamilies. The unstable nature of phasiine taxonomy discourages important research into their classification, life history and potential use in biological control. In hopes of resolving several longstanding taxonomic debates and encouraging future research into this important group of parasitoids, the first molecular systematic analysis of Phasiinae is presented, including 128 worldwide taxa (80 genera) and approximately 7.6 kb of nuclear data representing four genes. Special emphasis is placed on the resolution of taxonomically ambiguous groups. The resulting robustly supported phylogenetic trees [maximum‐likelihood (ML)/Bayesian] were used to trace the evolution of significant adaptive traits within Tachinidae and test hypotheses about the classification of Phasiinae. Subfamily placements of certain taxa are confidently resolved including Eutherini, Epigrimyiini, Litophasia Girschner within Dexiinae, and Strongygastrini and Parerigonini within Phasiinae. The members of tribe Phasiini are redistributed: Cistogaster Latreille, Clytiomya Rondani, Ectophasia Townsend, Eliozeta Rondani and Euclytia Townsend transferred to Gymnosomatini; Opesia Robineau‐Desvoidy to Strongygastrini; and Xysta Meigen to Xystini. Similarly, members of Parerigonini are treated as belonging to Parerigonini (Parerigone Brauer, Zambesomima Walker), Cylindromyiini (Australotachina Curran, Pygidimyia Crosskey, Neobrachelia Townsend) or new tribe Zitini (Zita Curran, Leverella Baranov). Penthosia van der Wulp is transferred from Cylindromyiini to Hermyini. Ancestral state reconstruction suggests that piercing structures used to insert eggs directly into host tissues have evolved separately in a number of groups, but have also been lost or reduced in several lineages. A single potentially unequivocal morphological synapomorphy of Phasiinae, an elongated medial plate of the hypandrium in males, is identified.

This published work has been registered in ZooBank,
http://zoobank.org/urn:lsid:zoobank.org:pub:8BE75122‐FC7C‐4809‐AAF7‐19575596EF78.
A new approach to geobarometry by combining fluid inclusion and clumped isotope thermometry in hydrothermal carbonates
Authors: Honlet Robin , Gasparrini Marta , Muchez Philippe , Swennen Rudy , John Cédric M.
Published: January 2018

This study presents a new approach to geobarometry by combining fluid inclusion and clumped isotope (Δ47) thermometry on carbonate minerals. The offset between homogenisation temperatures of primary fluid inclusions with known composition and Δ47 temperatures of the host mineral allows a direct estimation of the fluid pressure at the time of carbonate crystallisation. This new approach is illustrated via hydrothermal dolomite samples from the Variscan foreland fold‐and‐thrust belt in northern Spain. Clumped isotope analyses yield crystallisation temperatures (107–168°C) which are higher than homogenisation temperatures in corresponding samples (95–145°C). The calculated pressure values suggest that dolomitizing fluids were overpressured during formation of zebra dolomite textures, whereas lower pressures are obtained for dolomite cement from breccia textures. This new approach to geobarometry opens up the possibility of estimating the pressure of carbonate crystallisation and has potential applications in diagenesis, basin analysis, ore geology and tectonics.
Retrospective analysis of forward and reverse ABO typing discrepancies among patients and blood donors in a tertiary care hospital
Authors: Makroo R. N. , Kakkar B. , Agrawal S. , Chowdhry M. , Prakash B. , Karna P.
Published: January 2018

Objective

The aim of our study was to determine the incidence and causes of ABO typing discrepancies among patients and blood donors at our centre.

Background

An accurate interpretation of the ABO blood group of an individual is of utmost importance to ensure patient safety and good transfusion practices.

Methods

A retrospective observational study was carried out in the Department of Transfusion Medicine in our hospital from March 2013 to December 2015. Records of all patient and blood donor samples were retrieved and analysed for ABO typing discrepancies.

Results

In total, 135 853 patient and 62 080 donor samples were analysed for ABO typing discrepancies. The incidence among patients and blood donors was found to be 0·1% (138/135853) and 0·02% (14/62080), respectively. The mean age for patients and blood donors was 48·4 and 29·2 years, respectively. The most common cause of ABO typing discrepancies was due to cold autoantibodies among the patients (50·7%) and blood donors (57%) causing discrepant results in reverse typing. The various other causes of reverse typing discrepancies among patients were weak/missing antibody (25·4%), cold‐reacting alloantibody (4·3%), warm autoantibody (2·2%), anti‐A1 antibody (2·2%), Bombay phenotype (1·5%), transplantation (0·7%) and rouleaux (0·7%), whereas in blood donors, the causes were cold‐reacting antibody (7%) and weak antibody (7%). The major cause of forward typing discrepancies among patients (12·3%) and blood donors (29%) was ABO subgroups.

Conclusion

The resolution of ABO typing discrepancy is essential to minimise the chance of transfusion of ABO‐incompatible blood.
Treatment interruption in HIV‐positive patients followed up in Cameroon's antiretroviral treatment programme: individual and health care supply‐related factors (ANRS‐12288 EVOLCam survey)
Authors: Tong Christelle , Suzan‐Monti Marie , Sagaon‐Teyssier Luis , Mimi Mohamed , Laurent Christian , Maradan Gwenaëlle , Mengue Marie‐Thérèse , Spire Bruno , Kuaban Christopher , Vidal Laurent , Boyer Sylvie ,   the EVOL Cam group
Published: January 2018

Introduction

Decreasing international financial resources for HIV and increasing numbers of antiretroviral treatment (ART)‐treated patients may jeopardise treatment continuity in low‐income settings. Using data from the EVOLCam ANRS‐12288 survey, this study aimed to document the prevalence of unplanned treatment interruption for more than 2 consecutive days (TI>2d) and investigate the associated individual and health care supply‐related factors within the Cameroonian ART programme.

Methods

A cross‐sectional mixed methods survey was carried out between April and December 2014 in 19 HIV services of the Centre and Littoral regions. A multilevel logistic model was estimated on 1885 ART‐treated patients in these services to investigate factors of TI>2d in the past 4 weeks.

Results

Among the study population, 403 (21%) patients reported TI>2d. Patients followed up in hospitals reporting ART stock‐outs were more likely to report TI>2d while those followed up in the Littoral region, in medium‐ or small‐sized hospitals and in HIV services proposing financial support were at lower risk of TI>2d. The following individual factors were also associated with a lower risk of TI>2d: living in a couple, having children, satisfaction with attention provided by doctor, tuberculosis co‐infection and not having consulted a traditional healer.

Conclusions

Besides identifying individual factors of TI>2d, our study highlighted the role of health care supply‐related factors in shaping TI in Cameroon's ART programme, especially the deleterious effect of ART stock‐outs. Our results also suggest that the high proportion of patients reporting TI could jeopardise progress in the fight against HIV in the country, unless effective measures are quickly implemented like ensuring the continuity of ART supply.
Shank3‐deficient thalamocortical neurons show HCN channelopathy and alterations in intrinsic electrical properties
Authors: Zhu Mengye , Idikuda Vinay Kumar , Wang Jianbing , Wei Fusheng , Kumar Virang , Shah Nikhil , Waite Christopher B. , Liu Qinglian , Zhou Lei
Published: January 2018

Key points

Shank3 increases the HCN channel surface expression in heterologous expression systems.

Shank3Δ13–16 deficiency causes significant reduction in HCN2 expression and Ih current amplitude in thalamocortical (TC) neurons.

Shank3Δ13–16‐ but not Shank3Δ4–9‐deficient TC neurons share changes in basic electrical properties which are comparable to those of HCN2−/− TC neurons.

HCN channelopathy may critically mediate events downstream from Shank3 deficiency.

SHANK3 is a scaffolding protein that is highly enriched in excitatory synapses. Mutations in the SHANK3 gene have been linked to neuropsychiatric disorders especially the autism spectrum disorders. SHANK3 deficiency is known to cause impairments in synaptic transmission, but its effects on basic neuronal electrical properties that are more localized to the soma and proximal dendrites remain unclear. Here we confirmed that in heterologous expression systems two different mouse Shank3 isoforms, Shank3A and Shank3C, significantly increase the surface expression of the mouse hyperpolarization‐activated, cyclic‐nucleotide‐gated (HCN) channel. In Shank3Δ13–16 knockout mice, which lack exons 13–16 in the Shank3 gene (both Shank3A and Shank3C are removed) and display a severe behavioural phenotype, the expression of HCN2 is reduced to an undetectable level. The thalamocortical (TC) neurons from the ventrobasal (VB) complex of Shank3Δ13–16 mice demonstrate reduced Ih current amplitude and correspondingly increased input resistance, negatively shifted resting membrane potential, and abnormal spike firing in both tonic and burst modes. Impressively, these changes closely resemble those of HCN2−/− TC neurons but not of the TC neurons from Shank3Δ4–9 mice, which lack exons 4–9 in the Shank3 gene (Shank3C still exists) and demonstrate moderate behavioural phenotypes. Additionally, Shank3 deficiency increases the ratio of excitatory/inhibitory balance in VB neurons but has a limited impact on the electrical properties of connected thalamic reticular (RTN) neurons. These results provide new understanding about the role of HCN channelopathy in mediating detrimental effects downstream from Shank3 deficiency.
Kinematic Analysis of a Novel Kinematically Redundant Spherical Parallel Manipulator
Authors: Landuré Jérôme , Gosselin Clément
Published: January 2018

This paper introduces a new architecture of spherical parallel robot which significantly extends the workspace when compared to existing architectures. To this end, the singularity locus is studied and the design parameters are chosen so as to confine the singularities to areas already limited by other constraints such as mechanical interferences. First, the architecture of the spherical redundant robot is presented and the Jacobian matrices are derived. Afterwards, the analysis of the singularities is addressed from a geometric point of view, which yields a description of the singularity locus expressed as a function of the architectural parameters. Then, the results are applied to an example set of architectural parameters, which are chosen in order to illustrate the advantages of the redundant architecture over current designs in terms of workspace.
Rigidly Foldable Quadrilateral Meshes From Angle Arrays
Authors: Lang Robert J. , Howell Larry
Published: January 2018

We present a design technique for generating rigidly foldable quadrilateral meshes (RFQMs), taking as input four arrays of direction angles and fold angles for horizontal and vertical folds. By starting with angles, rather than vertex coordinates, and enforcing the fold-angle multiplier condition at each vertex, it is possible to achieve arbitrarily large and complex panel arrays that flex from unfolded to flatly folded with a single degree-of-freedom (DOF). Furthermore, the design technique is computationally simple, reducing for some cases to a simple linear-programming problem. The resulting mechanisms have applications in architectural facades, furniture design, and more.
A Prismatic-Revolute-Revolute Joint Hand for Grasping From Unmanned Aerial Vehicles and Other Minimally Constrained Vehicles
Authors: Backus Spencer B. , Dollar Aaron M.
Published: January 2018

Here, we present the design, fabrication, and evaluation of a prismatic-revolute-revolute joint hand called the model B that we developed for grasping from ungrounded vehicles. This hand relies on a prismatic proximal joint followed by revolute distal joints in each finger and is actuated by a single motor- and a tendon-based underactuated transmission. We evaluate this design's grasping capabilities both when fully constrained by a robotic arm and when minimally constrained and evaluate its performance in terms of general grasping capabilities and suitability for aerial grasping applications. The evaluation shows that the model B can securely grasp a wide range of objects using a wrap grasp due to the prismatic-revolute-revolute joint finger kinematics. We also show that the prismatic proximal joints and between finger coupling allows the hand to grasp objects under large positional uncertainty without exerting large reaction forces on the object or host vehicle.
An Elastoplastic Finite Element Study of Displacement-Controlled Fretting in a Plane-Strain Cylindrical Contact
Authors: Yang Huaidong , Green Itzhak
Published: January 2018

This work presents a finite element study of a two-dimensional (2D) plane strain fretting model of a half cylinder in contact with a flat block under oscillatory tangential loading. The two bodies are deformable and are set to the same material properties (specifically steel), however, because the results are normalized, they can characterize a range of contact scales (micro to macro), and are applicable for ductile material pairs that behave in an elastic-perfectly plastic manner. Different coefficients of friction (COFs) are used in the interface. This work finds that the edges of the contacting areas experience large von Mises stresses along with significant residual plastic strains, while pileup could also appear there when the COFs are sufficiently large. In addition, junction growth is investigated, showing a magnitude that increases with the COF, while the rate of growth stabilization decreases with the COF. The fretting loop (caused by the tangential force during the fretting motion) for the initial few cycles of loading is generated, and it compares well with reported experimental results. The effects of boundary conditions are also discussed where a prestressed compressed block is found to improve (i.e., reduce) the magnitude of the plastic strain compared to an unstressed block.
Real-Time Variable Geometry Triaxial Gas Turbine Model for Hardware-in-the-Loop Simulation Experiments
Authors: Wang Tao , Tian Yong-Sheng , Yin Zhao , Zhang Da-Yue , Ma Ming-Ze , Gao Qing , Tan Chun-Qing
Published: January 2018

This paper proposes a hybrid method (HMRC) comprised of a radial basis function (RBF) neural net algorithm and component-level modeling method (CMM) as a real-time simulation model for triaxial gas turbines with variable power turbine guide vanes in matlab/simulink. The sample size is decreased substantially after analyzing the relationship between high and low pressure shaft rotational speeds under dynamic working conditions, which reduces the computational burden of the simulation. The effects of the power turbine rotational speed on overall performance are also properly accounted for in the model. The RBF neural net algorithm and CMM are used to simulate the gas generator and power turbine working conditions, respectively, in the HMRC. The reliability and accuracy of both the traditional single CMM model (SCMM) and HMRC model are verified using gas turbine experiment data. The simulation models serve as a controlled object to replace the real gas turbine in a hardware-in-the-loop simulation experiment. The HMRC model shows better real-time performance than the traditional SCMM model, suggesting that it can be readily applied to hardware-in-the-loop simulation experiments.
Modeling Particle Spray and Capture Efficiency for Direct Laser Deposition Using a Four Nozzle Powder Injection System
Authors: Katinas Christopher , Shang Weixiao , Shin Yung C. , Chen Jun
Published: January 2018

Powder capture efficiency is indicative of the amount of material that is added to the substrate during laser additive manufacturing (AM) processes, and thus, being able to predict capture efficiency provides capability of predictive modeling during such processes. The focus of the work presented in this paper is to create a numerical model to understand particle trajectories and velocities, which in turn allows for the prediction of capture efficiency. To validate the numerical model, particle tracking velocimetry (PTV) experiments at two powder flow rates were conducted on free stream particle spray to track individual particles such that particle concentration and velocity fields could be obtained. Results from the free stream comparison showed good agreement to the trends observed in experimental data and were subsequently used in a direct laser deposition (DLD) simulation to assess capture efficiency and temperature profile at steady-state. The simulation was validated against a single track deposition experiment and showed proper correlation of the free surface geometry, molten pool boundary, heat affected zone boundary, and capture efficiency.
Dependence of the subharmonic signal from contrast agent microbubbles on ambient pressure: A theoretical analysis
Authors: Jiménez-Fernández J.
Published: January 2018

This paper investigates the dependence of the subharmonic response in a signal scattered by contrast agent microbubbles on ambient pressure to provide quantitative estimations of local blood pressure. The problem is formulated by assuming a gas bubble encapsulated by a shell of finite thickness with dynamic behavior modeled by a nonlinear viscoelastic constitutive equation. For ambient overpressure compatible with the clinical range, the acoustic pressure intervals where the subharmonic signal may be detected (above the threshold for the onset and below the limit value for the first chaotic transition) are determined. The analysis shows that as the overpressure is increased, all harmonic components are displaced to higher frequencies. This displacement is significant for the subharmonic of order 1/2 and explains the increase or decrease in the subharmonic amplitude with ambient pressure described in previous works. Thus, some questions related to the monotonic dependence of the subharmonic amplitude on ambient pressure are clarified. For different acoustic pressures, quantitative conditions for determining the intervals where the subharmonic amplitude is a monotonic or non-monotonic function of the ambient pressure are provided. Finally, the influence of the ambient pressure on the subharmonic resonance frequency is analyzed.
Germination, Outgrowth, and Vegetative-Growth Kinetics of Dry-Heat-Treated Individual Spores of Bacillus Species
Authors: He Lin , Chen Zhan , Wang Shiwei , Wu Muying , Setlow Peter , Li Yong-qing
Published: January 2018

DNA damage kills dry-heated spores of
Bacillus subtilis
, but dry-heat-treatment effects on spore germination and outgrowth have not been studied. This is important, since if dry-heat-killed spores germinate and undergo outgrowth, toxic proteins could be synthesized. Here, Raman spectroscopy and differential interference contrast microscopy were used to study germination and outgrowth of individual dry-heat-treated
B. subtilis
and
Bacillus megaterium
spores. The major findings in this work were as follows: (i) spores dry-heat-treated at 140°C for 20 min lost nearly all viability but retained their Ca
2+
-dipicolinic acid (CaDPA) depot; (ii) in most cases, dry-heat treatment increased the average times and variability of all major germination events in
B. subtilis
spore germination with nutrient germinants or CaDPA, and in one nutrient germination event with
B. megaterium
spores; (iii)
B. subtilis
spore germination with dodecylamine, which activates the spore CaDPA release channel, was unaffected by dry-heat treatment; (iv) these results indicate that dry-heat treatment likely damages spore proteins important in nutrient germinant recognition and cortex peptidoglycan hydrolysis, but not CaDPA release itself; and (v) analysis of single spores incubated on nutrient-rich agar showed that while dry-heat-treated spores that are dead can complete germination, they cannot proceed into outgrowth and thus not to vegetative growth. The results of this study provide new information on the effects of dry heat on bacterial spores and indicate that dry-heat sterilization regimens should produce spores that cannot outgrow and thus cannot synthesize potentially dangerous proteins.

IMPORTANCE
Much research has shown that high-temperature dry heat is a promising means for the inactivation of spores on medical devices and spacecraft decontamination. Dry heat is known to kill
Bacillus subtilis
spores by DNA damage. However, knowledge about the effects of dry-heat treatment on spore germination and outgrowth is limited, especially at the single spore level. In the current work, Raman spectroscopy and differential interference contrast microscopy were used to analyze CaDPA levels in and kinetics of nutrient- and non-nutrient germination of multiple individual dry-heat-treated
B. subtilis
and
Bacillus megaterium
spores that were largely dead. The outgrowth and subsequent cell division of these germinated but dead dry-heat-treated spores were also examined. The knowledge obtained in this study will help understand the effects of dry heat on spores both on Earth and in space, and indicates that dry heat can be safely used for sterilization purposes.
Parainfluenza Virus Infection Sensitizes Cancer Cells to DNA-Damaging Agents: Implications for Oncolytic Virus Therapy
Authors: Fox Candace R. , Parks Griffith D.
Published: January 2018

A parainfluenza virus 5 (PIV5) with mutations in the P/V gene (P/V-CPI
−
) is restricted for spread in normal cells but not in cancer cells
in vitro
and is effective at reducing tumor burdens in mouse model systems. Here we show that P/V-CPI
−
infection of HEp-2 human laryngeal cancer cells results in the majority of the cells dying, but unexpectedly, over time, there is an emergence of a population of cells that survive as P/V-CPI
−
persistently infected (PI) cells. P/V-CPI
−
PI cells had elevated levels of basal caspase activation, and viability was highly dependent on the activity of cellular inhibitor-of-apoptosis proteins (IAPs) such as Survivin and XIAP. In challenge experiments with external inducers of apoptosis, PI cells were more sensitive to cisplatin-induced DNA damage and cell death. This increased cisplatin sensitivity correlated with defects in DNA damage signaling pathways such as phosphorylation of Chk1 and translocation of damage-specific DNA binding protein 1 (DDB1) to the nucleus. Cisplatin-induced killing of PI cells was sensitive to the inhibition of wild-type (WT) p53-inducible protein 1 (WIP1), a phosphatase which acts to terminate DNA damage signaling pathways. A similar sensitivity to cisplatin was seen with cells during acute infection with P/V-CPI
−
as well as during acute infections with WT PIV5 and the related virus human parainfluenza virus type 2 (hPIV2). Our results have general implications for the design of safer paramyxovirus-based vectors that cannot establish PI as well as the potential for combining chemotherapy with oncolytic RNA virus vectors.

IMPORTANCE
There is intense interest in developing oncolytic viral vectors with increased potency against cancer cells, particularly those cancer cells that have gained resistance to chemotherapies. We have found that infection with cytoplasmically replicating parainfluenza virus can result in increases in the killing of cancer cells by agents that induce DNA damage, and this is linked to alterations to DNA damage signaling pathways that balance cell survival versus death. Our results have general implications for the design of safer paramyxovirus-based vectors that cannot establish persistent infection, the repurposing of drugs that target cellular IAPs as antivirals, and the combined use of DNA-damaging chemotherapy agents in conjunction with oncolytic RNA virus vectors.
Attenuation of Simian Varicella Virus Infection by Enhanced Green Fluorescent Protein in Rhesus Macaques
Authors: Mahalingam Ravi , Kaufer Benedikt B. , Ouwendijk Werner J. D. , Verjans Georges M. G. M. , Coleman Colin , Hunter Meredith , Das Arpita , Palmer Brent E. , Clambey Eric , Nagel Maria A. , Traina-Dorge Vicki
Published: January 2018

Simian varicella virus (SVV), the primate counterpart of varicella-zoster virus, causes varicella (chickenpox), establishes latency in ganglia, and reactivates to produce zoster. We previously demonstrated that a recombinant SVV expressing enhanced green fluorescent protein (rSVV.eGFP) is slightly attenuated both in culture and in infected monkeys. Here, we generated two additional recombinant SVVs to visualize infected cells
in vitro
and
in vivo
. One harbors eGFP fused to the N terminus of open reading frame 9 (ORF9) (rSVV.eGFP-2a-ORF9), and another harbors eGFP fused to the C terminus of ORF66 (rSVV.eGFP-ORF66). Both recombinant viruses efficiently expressed eGFP in cultured cells. Both recombinant SVV infections in culture were comparable to that of wild-type SVV (SVV.wt). Unlike SVV.wt, eGFP-tagged SVV did not replicate in rhesus cells in culture. Intratracheal (i.t.) or i.t. plus intravenous (i.v.) inoculation of rhesus macaques with these new eGFP-tagged viruses resulted in low viremia without varicella rash, although SVV DNA was abundant in bronchoalveolar lavage (BAL) fluid at 10 days postinoculation (dpi). SVV DNA was also found in trigeminal ganglia of one monkey inoculated with rSVV.eGFP-ORF66. Intriguingly, a humoral response to both SVV and eGFP was observed. In addition, monkeys inoculated with the eGFP-expressing viruses were protected from superinfection with SVV.wt, suggesting that the monkeys had mounted an efficient immune response. Together, our results show that eGFP expression could be responsible for their reduced pathogenesis.

IMPORTANCE
SVV infection in nonhuman primates has served as an extremely useful animal model to study varicella-zoster virus (VZV) pathogenesis. eGFP-tagged viruses are a great tool to investigate their pathogenesis. We constructed and tested two new recombinant SVVs with eGFP inserted into two different locations in the SVV genome. Both recombinant SVVs showed robust replication in culture but reduced viremia compared to that with SVV.wt during primary infection in rhesus macaques. Our results indicate that conclusions on eGFP-tagged viruses based on
in vitro
results should be handled with care, since eGFP expression could result in attenuation of the virus.
Inter-rater reliability in the Paediatric Observation Priority Score (POPS)
Authors: Langton Lisa , Bonfield Adam , Roland Damian
Published: January 2018

Objective

The primary objective of this study was to determine the level of inter-rater reliability between nursing staff for the Paediatric Observation Priority Score (POPS).

Design

Retrospective observational study.

Setting

Single-centre paediatric emergency department.

Participants

12 participants from a convenience sample of 21 nursing staff.

Interventions

Participants were shown video footage of three pre-recorded paediatric assessments and asked to record their own POPS for each child. The participants were blinded to the original, in-person POPS. Further data were gathered in the form of a questionnaire to determine the level of training and experience the candidate had using the POPS score prior to undertaking this study.

Main outcome measures

Inter-rater reliability among participants scoring of the POPS.

Results

Overall kappa value for case 1 was 0.74 (95% CI 0.605 to 0.865), case 2 was 1 (perfect agreement) and case 3 was 0.66 (95% CI 0.58 to 0.744).

Conclusion

This study suggests there is good inter-rater reliability between different nurses’ use of POPS in assessing sick children in the emergency department.
What are the sources of stress and distress for general practitioners working in England? A qualitative study
Authors: Riley Ruth , Spiers Johanna , Buszewicz Marta , Taylor Anna Kathryn , Thornton Gail , Chew-Graham Carolyn Anne
Published: January 2018

Objectives

This paper reports the sources of stress and distress experienced by general practitioners (GP) as part of a wider study exploring the barriers and facilitators to help-seeking for mental illness and burnout among this medical population.

Design

Qualitative study using in-depth interviews with 47 GP participants. The interviews were audio-recorded, transcribed, anonymised and imported into NVivo V.11 to facilitate data management. Data were analysed using a thematic analysis employing the constant comparative method.

Setting

England.

Participants

A purposive sample of GP participants who self-identified as: (1) currently living with mental distress, (2) returning to work following treatment, (3) off sick or retired early as a result of mental distress or (4) without experience of mental distress. Interviews were conducted face-to-face or over the telephone.

Results

The key sources of stress/distress related to: (1) emotion work—the work invested and required in managing and responding to the psychosocial component of GPs’ work, and dealing with abusive or confrontational patients; (2) practice culture—practice dynamics and collegial conflict, bullying, isolation and lack of support; (3) work role and demands—fear of making mistakes, complaints and inquests, revalidation, appraisal, inspections and financial worries.

Conclusion

In addition to addressing escalating workloads through the provision of increased resources, addressing unhealthy practice cultures is paramount. Collegial support, a willingness to talk about vulnerability and illness, and having open channels of communication enable GPs to feel less isolated and better able to cope with the emotional and clinical demands of their work. Doctors, including GPs, are not invulnerable to the clinical and emotional demands of their work nor the effects of divisive work cultures—culture change and access to informal and formal support is therefore crucial in enabling GPs to do their job effectively and to stay well.
Is placebo response in antidepressant trials rising or not? A reanalysis of datasets to conclude this long-lasting controversy
Authors: Furukawa Toshi A , Cipriani Andrea , Leucht Stefan , Atkinson Lauren Z , Ogawa Yusuke , Takeshima Nozomi , Hayasaka Yu , Chaimani Anna , Salanti Georgia
Published: January 2018

It had long been believed that placebo response rates in antidepressant trials have been increasing and that they were responsible for rising numbers of so-called failed antidepressant trials. Two recent systematic reviews examined this issue and reached completely opposite findings. Furukawa and colleagues in a paper published in 2016 found that the placebo response rates are stable since 1991 and the apparent increase up to 2000 was confounded by changes in trial design features. By contrast, Khan and colleagues more recently concluded that placebo response rates had grown steadily in the past 30 years. The two reviews differed in the datasets they used, definitions of placebo response and statistical analyses. In this perspective article, we examined if such differences were responsible for the two reviews’ contrasting conclusions. Our reanalyses confirmed our previous results. We found that in any dataset and for any placebo response definition, there was no increase in placebo response over the years when the analysis was adjusted for the confounders related to study design features or when it was limited to studies published after 1990s. We conclude that placebo response in antidepressant trials has remained stable for the past 25 years, during which time the large majority of the studies have come to share similar design features.
Survival in relation to multimorbidity patterns in older adults in primary care in Barcelona, Spain (2010–2014): a longitudinal study based on electronic health records
Authors: Ibarra-Castillo Carolina , Guisado-Clavero Marina , Violan-Fors Concepció , Pons-Vigués Mariona , López-Jiménez Tomàs , Roso-Llorach Albert
Published: January 2018

Background

Several studies have analysed the characteristics of multimorbidity patterns but none have evaluated the relationship with survival. The purpose of this study was to compare survival across older adults with different chronic multimorbidity patterns (CMPs).

Methods

Prospective longitudinal observational study using electronic health records for 190 108 people aged ≥65 years in Barcelona, Spain (2009–2014). CMPs were identified by cluster analysis. Mortality rates were estimated using the Catalan population structure and individual time at risk. Survival according to CMP (Cox regression) was analysed using hazard ratios (HRs) and 95% confidence intervals (CIs) with stratification by sex and age group (65–79, 80–94) and adjustment for age at onset, deprivation index, number of chronic conditions and invoiced drugs.

Results

The highest mortality rates were observed in men, adults aged 80–94 years, socially disadvantaged quintiles and people prescribed more drugs and with fewer conditions. Using the musculoskeletal pattern as the reference category, men with the digestive-respiratory pattern had a higher risk of death, with adjusted HRs of 6.16 (95% CI 5.37 to 7.06) in the 65–79 age group and 2.62 (95% CI 2.31 to 2.97) in the 80–94 age group. In women, the cardiovascular pattern was associated with the highest risk, with adjusted HRs of 6.34 (95% CI 5.28 to 7.61) in the 65–79 age group and 3.05 (95% CI 2.73 to 3.41) in the 80–94 age group. These patterns were also associated with the highest mortality rates.

Conclusions

Mortality and survival vary according to CMPs in older adults stratified by sex and age. Our findings are useful for guiding the design and implementation of clinical management strategies.
Role of germline aberrations affecting CTNNA1, MAP3K6 and MYD88 in gastric cancer susceptibility
Authors: Weren Robbert D A , van der Post Rachel S , Vogelaar Ingrid P , van Krieken J Han , Spruijt Liesbeth , Lubinski Jan , Jakubowska Anna , Teodorczyk Urszula , Aalfs Cora M , van Hest Liselotte P , Oliveira Carla , Kamping Eveline J , Schackert Hans K , Ranzani Guglielmina N , Gómez García Encarna B , Hes Frederik J , Holinski-Feder Elke , Genuardi Maurizio , Ausems Margreet G E M , Sijmons Rolf H , Wagner Anja , van der Kolk Lizet E , Cats Annemieke , Bjørnevoll Inga , Hoogerbrugge Nicoline , Ligtenberg Marjolijn J L
Published: January 2018

Background

In approximately 10% of all gastric cancer (GC) cases, a heritable cause is suspected. A subset of these cases have a causative germline CDH1 mutation; however, in most cases the cause remains unknown. Our objective was to assess to what extent these remaining cases may be explained by germline mutations in the novel candidate GC predisposing genes CTNNA1, MAP3K6 or MYD88.

Methods

We sequenced a large cohort of unexplained young and/or familial patients with GC (n=286) without a CDH1germline mutation for germline variants affecting CTNNA1, MAP3K6 and MYD88 using a targeted next-generation sequencing approach based on single-molecule molecular inversion probes.

Results

Predicted deleterious germline variants were not encountered in MYD88, but recurrently observed in CTNNA1 (n=2) and MAP3K6 (n=3) in our cohort of patients with GC. In contrast to deleterious variants in CTNNA1, deleterious variants in MAP3K6 also occur frequently in the general population.

Conclusions

Based on our results MAP3K6 should no longer be considered a GC predisposition gene, whereas deleterious CTNNA1 variants are confirmed as an infrequent cause of GC susceptibility. Biallelic MYD88 germline mutations are at most a very rare cause of GC susceptibility as no additional cases were identified.
Fabry Disease: prevalence of affected males and heterozygotes with pathogenic GLA mutations identified by screening renal, cardiac and stroke clinics, 1995–2017
Authors: Doheny Dana , Srinivasan Ram , Pagant Silvere , Chen Brenden , Yasuda Makiko , Desnick Robert J
Published: January 2018

Background

Fabry Disease (FD), an X linked lysosomal storage disease due to pathogenic α-galactosidase A (GLA) mutations, results in two major subtypes, the early-onset Type 1 ‘Classic’ and the Type 2 ‘Later-Onset’ phenotypes. To identify previously unrecognised patients, investigators screened cardiac, renal and stroke clinics by enzyme assays. However, some screening studies did not perform confirmatory GLA mutation analyses, and many included recently recognised ‘benign/likely-benign’ variants, thereby inflating prevalence estimates.

Methods

Online databases were searched for all FD screening studies in high-risk clinics (1995–2017). Studies reporting GLA mutations were re-analysed for pathogenic mutations, sex and phenotype. Phenotype-specific and sex-specific prevalence rates were determined.

Results

Of 67 studies, 63 that screened 51363patients (33943M and 17420F) and provided GLA mutations were reanalysed for disease-causing mutations. Of reported GLA mutations, benign variants occurred in 47.9% of males and 74.1% of females. The following were the revised prevalence estimates: among 36820 (23954M and 12866F) haemodialysis screenees, 0.21% males and 0.15% females; among 3074 (2031M and 1043F) renal transplant screenees, 0.25% males and no females; among 5491 (4054M and 1437F) cardiac screenees, 0.94% males and 0.90% females; and among 5978 (3904M and 2074F) stroke screenees, 0.13% males and 0.14% females. Among male and female screenees with pathogenic mutations, the type 1 Classic phenotype was predominant (~60%), except more male cardiac patients (75%) had type 2 Later-Onset phenotype.

Conclusions

Compared with previous findings, reanalysis of 63 studies increased the screenee numbers (~3.4-fold), eliminated 20 benign/likely benign variants, and provided more accurate sex-specific and phenotype-specific prevalence estimates, ranging from ~0.13% of stroke to ~0.9% of cardiac male or female screenees.
The Casper Stent System for carotid artery stenosis
Authors: Mutzenbach Sebastian J , Millesi Katharina , Roesler Cornelia , Broussalis Erasmia , Pikija Slaven , Sellner Johann , Machegger Lukas , Griessenauer Christoph J , Killer-Oberpfalzer Monika
Published: January 2018

Purpose

To report the results of a retrospective analysis of prospectively collected data evaluating the safety and efficacy of a double layer stent engineered for carotid artery occlusive disease.

Methods

Between January 2014 and February 2017, 138 patients (25.4% women; median age 71 years) underwent Casper stent implantation for carotid artery stenosis. Eligibility criteria included stenosis >70% of vessel diameter (or >50% diameter with ulceration) in symptomatic patients or asymptomatic patients with >80% stenosis at the carotid bifurcation or in the proximal internal carotid artery. For all procedures, a distal embolic protection device was used. The primary endpoint was the rate of 90 day major adverse neurological events, defined as minor stroke, major stroke, or death by independent neurological assessment.

Results

Stent deployment was completed successfully in all cases without documented technical failure. There were no adverse neurological events or mortalities within 90 days. One thromboembolic occlusion of a small distal branch of the anterior cerebral artery occurred during the procedure and resolved with systemic recombinant tissue plasminogen activator administration. New ischemic lesions, all clinically silent, were seen in 6.5% of patients on post-procedure cerebral MRI.

Conclusion

The Casper carotid stent demonstrated safety and efficacy in the treatment of carotid stenosis, with no technical failures and no adverse neurological events seen throughout the 90 day follow-up period. Its double layer structure seems to combine adequate plaque scaffolding with high vessel adaptability.