bluuebunny/crossref_metadata_2025 huggingface dataset:
Species Specificity of Vaccinia Virus Complement Control Protein for the Bovine Classical Pathway Is Governed Primarily by Direct Interaction of Its Acidic Residues with Factor I
Authors: Kumar Jitendra , Yadav Viveka Nand , Phulera Swastik , Kamble Ashish , Gautam Avneesh Kumar , Panwar Hemendra Singh , Sahu Arvind
Published: July 2017

Poxviruses display species tropism—variola virus is a human-specific virus, while vaccinia virus causes repeated outbreaks in dairy cattle. Consistent with this, variola virus complement regulator SPICE (smallpox inhibitor of complement enzymes) exhibits selectivity in inhibiting the human alternative complement pathway and vaccinia virus complement regulator VCP (vaccinia virus complement control protein) displays selectivity in inhibiting the bovine alternative complement pathway. In the present study, we examined the species specificity of VCP and SPICE for the classical pathway (CP). We observed that VCP is ∼43-fold superior to SPICE in inhibiting bovine CP. Further, functional assays revealed that increased inhibitory activity of VCP for bovine CP is solely due to its enhanced cofactor activity, with no effect on decay of bovine CP C3-convertase. To probe the structural basis of this specificity, we utilized single- and multi-amino-acid substitution mutants wherein 1 or more of the 11 variant VCP residues were substituted in the SPICE template. Examination of these mutants for their ability to inhibit bovine CP revealed that E108, E120, and E144 are primarily responsible for imparting the specificity and contribute to the enhanced cofactor activity of VCP. Binding and functional assays suggested that these residues interact with bovine factor I but not with bovine C4(H2O) (a moiety conformationally similar to C4b). Mapping of these residues onto the modeled structure of bovine C4b-VCP-bovine factor I supported the mutagenesis data. Taken together, our data help explain why the vaccine strain of vaccinia virus was able to gain a foothold in domesticated animals.

IMPORTANCEVaccinia virus was used for smallpox vaccination. The vaccine-derived virus is now circulating and causing outbreaks in dairy cattle in India and Brazil. However, the reason for this tropism is unknown. It is well recognized that the virus is susceptible to neutralization by the complement classical pathway (CP). Because the virus encodes a soluble complement regulator, VCP, we examined whether this protein displays selectivity in targeting bovine CP. Our data show that it does exhibit selectivity in inhibiting the bovine CP and that this is primarily determined by its amino acids E108, E120, and E144, which interact with bovine serine protease factor I to inactivate bovine C4b—one of the two subunits of CP C3-convertase. Of note, the variola complement regulator SPICE contains positively charged residues at these positions. Thus, these variant residues in VCP help enhance its potency against the bovine CP and thereby the fitness of the virus in cattle.
UV Radiation Activates Toll-Like Receptor 9 Expression in Primary Human Keratinocytes, an Event Inhibited by Human Papillomavirus 38 E6 and E7 Oncoproteins
Authors: Pacini Laura , Ceraolo Maria Grazia , Venuti Assunta , Melita Giusi , Hasan Uzma A. , Accardi Rosita , Tommasino Massimo
Published: July 2017

Several lines of evidence indicate that cutaneous human papillomavirus (HPV) types belonging to the beta genus of the HPV phylogenetic tree synergize with UV radiation in the development of skin cancer. Accordingly, the E6 and E7 oncoproteins from some beta HPV types are able to deregulate pathways related to immune response and cellular transformation. Toll-like receptor 9 (TLR9), in addition to playing a role in innate immunity, has been shown to be involved in the cellular stress response. Using primary human keratinocytes as experimental models, we have shown that UV irradiation (and other cellular stresses) activates TLR9 expression. This event is closely linked to p53 activation. Silencing the expression of p53 or deleting its encoding gene affected the activation of TLR9 expression after UV irradiation. Using various strategies, we have also shown that the transcription factors p53 and c-Jun are recruited onto a specific region of the TLR9 promoter after UV irradiation. Importantly, the E6 and E7 oncoproteins from beta HPV38, by inducing the accumulation of the p53 antagonist ΔNp73α, prevent the UV-mediated recruitment of these transcription factors onto the TLR9 promoter, with subsequent impairment of TLR9 gene expression. This study provides new insight into the mechanism that mediates TLR9 upregulation in response to cellular stresses. In addition, we show that HPV38 E6 and E7 are able to interfere with this mechanism, providing another explanation for the possible cooperation of beta HPV types with UV radiation in skin carcinogenesis.

IMPORTANCE
Beta HPV types have been suggested to act as cofactors in UV-induced skin carcinogenesis by altering several cellular mechanisms activated by UV radiation. We show that the expression of TLR9, a sensor of damage-associated molecular patterns produced during cellular stress, is activated by UV radiation in primary human keratinocytes (PHKs). Two transcription factors known to be activated by UV radiation, p53 and c-Jun, play key roles in UV-activated TLR9 expression. The E6 and E7 oncoproteins from beta HPV38 strongly inhibit UV-activated TLR9 expression by preventing the recruitment of p53 and c-Jun to the TLR9 promoter. Our findings provide additional support for the role that beta HPV types play in skin carcinogenesis by preventing activation of specific pathways upon exposure of PHKs to UV radiation.
Intracellular Colocalization of Influenza Viral RNA and Rab11A Is Dependent upon Microtubule Filaments
Authors: Nturibi Eric , Bhagwat Amar R. , Coburn Stefanie , Myerburg Mike M. , Lakdawala Seema S.
Published: July 2017

Influenza A virus (IAV) consists of eight viral RNA (vRNA) segments that are replicated in the host cell nucleus and transported to the plasma membrane for packaging into progeny virions. We have previously proposed a model where subcomplexes of vRNA are exported from the nucleus and assembled en route to the plasma membrane. However, the role of host cytoskeletal proteins in the cytoplasmic assembly of IAV vRNA segments remains unknown. Previous studies have suggested that IAV vRNA segments are transported via Rab11A-containing recycling endosomes (RE) and use both microtubules (MT) and actin. Rab11A RE transport primarily along MT; therefore, investigation of the role of MT in vRNA assembly is warranted. We explored the role of MT in vRNA assembly and replication by using multiple IAV strains in various cell types, including primary human airway epithelial cells. We observed that Rab11A localization was altered in the presence of MT-depolymerizing drugs, but growth of IAV in all of the cell types tested was unchanged. Fluorescent
in situ
hybridization was performed to determine the role of MT in the assembly of multiple vRNA segments. Unexpectedly, we found that vRNA-vRNA association in cytoplasmic foci was independent of MT. Given the disparity of localization between Rab11A and vRNA segments in the absence of intact MT filaments, we analyzed the three-dimensional spatial relationship between Rab11A and vRNA in the cytoplasm of infected cells. We found that Rab11A and vRNA colocalization is dependent upon dynamic MT filaments. Taken together, our data suggest that cytoplasmic transport of influenza vRNA may include a Rab11A RE-independent mechanism.

IMPORTANCE
IAV infections cause a large public health burden through seasonal epidemics and sporadic pandemics. Pandemic IAVs emerge through reassortment of vRNA in animal or human hosts. Elucidation of the mechanism of intracellular dynamics of IAV assembly is necessary to understand reassortment. Our results describing the role of MT in vRNA transport and assembly expand upon previous studies characterizing vRNA assembly. This study is the first to assess the role of MT in influenza virus replication in human bronchial airway epithelial cells. In addition, we present novel data on the role of MT in facilitating the association between distinct vRNA segments. Interestingly, our results suggest that progressive assembly of vRNA segments may be cell type dependent and that vRNA may be transported through the cytoplasm without Rab11A RE in the absence of intact MT. These results enhance our understanding of vRNA assembly and the role of cytoskeletal proteins in that process.
The Effector TepP Mediates Recruitment and Activation of Phosphoinositide 3-Kinase on Early Chlamydia trachomatis Vacuoles
Authors: Carpenter Victoria , Chen Yi-Shan , Dolat Lee , Valdivia Raphael H.
Published: July 2017

This article shows that chlamydia recruit PI3K, an enzyme important for host cell survival and internal membrane functions, to the pathogens inside cells by secreting a scaffolding protein called TepP. TepP enhances
Chlamydia
replication and dampens the activation of immune responses.
Functioning metastatic paraganglioma of the urinary bladder in a 10-year-old child
Authors: El Alayli Alaeddine , El Amine Mohammad Ali , El Hout Yaser , Naffaa Lena
Published: July 2017

Paragangliomas of the urinary bladder are very rare tumours representing less than 1% of bladder tumours. Preoperative diagnosis is essential to avoid perioperative complications related to catecholamine release. A high index of suspicion should be maintained when the classical symptoms of voiding-related paroxysms of headache, palpitation and dizziness are present. We present a rare case of malignant paraganglioma of the urinary bladder in a 10-year-old boy. The patient had the classic presentation. We review the radiological diagnostic tools and findings of this rare entity.
Sedentary time in older men and women: an international consensus statement and research priorities
Authors: Dogra Shilpa , Ashe Maureen C , Biddle Stuart J H , Brown Wendy J , Buman Matthew P , Chastin Sebastien , Gardiner Paul A , Inoue Shigeru , Jefferis Barbara J , Oka Koichiro , Owen Neville , Sardinha Luís B , Skelton Dawn A , Sugiyama Takemi , Copeland Jennifer L
Published: July 2017

Sedentary time is a modifiable determinant of poor health, and in older adults, reducing sedentary time may be an important first step in adopting and maintaining a more active lifestyle. The primary purpose of this consensus statement is to provide an integrated perspective on current knowledge and expert opinion pertaining to sedentary behaviour in older adults on the topics of measurement, associations with health outcomes, and interventions. A secondary yet equally important purpose is to suggest priorities for future research and knowledge translation based on gaps identified. A five-step Delphi consensus process was used. Experts in the area of sedentary behaviour and older adults (n=15) participated in three surveys, an in-person consensus meeting, and a validation process. The surveys specifically probed measurement, health outcomes, interventions, and research priorities. The meeting was informed by a literature review and conference symposium, and it was used to create statements on each of the areas addressed in this document. Knowledge users (n=3) also participated in the consensus meeting. Statements were then sent to the experts for validation. It was agreed thatself-reporttools need to be developed for understanding the context in which sedentary time is accumulated. Forhealth outcomes, it was agreed that the focus of sedentary time research in older adults needs to include geriatric-relevant health outcomes, that there is insufficient evidence to quantify the dose–response relationship, that there is a lack of evidence on sedentary time from older adults in assisted facilities, and that evidence on the association between sedentary time and sleep is lacking. Forinterventions,research is needed to assess the impact that reducing sedentary time, or breaking up prolonged bouts of sedentary time has on geriatric-relevant health outcomes. Research priorities listed for each of these areas should be considered by researchers and funding agencies.

This consensus statement has been endorsed by the following societies: Academy of Geriatric Physical Therapy, Exercise & Sports Science Australia, Canadian Centre for Activity and Aging, Society of Behavioral Medicine, and the National Centre for Sport and Exercise Medicine.
Ten years transmission of the new variant of Chlamydia trachomatis in Sweden: prevalence of infections and associated complications
Authors: Dahlberg Jenny , Hadad Ronza , Elfving Karin , Larsson Inger , Isaksson Jenny , Magnuson Anders , Fredlund Hans , Unemo Magnus , Herrmann Bjőrn
Published: July 2017

Objectives

In 2006, a new variant of Chlamydia trachomatis (nvCT) was discovered in Sweden. It has a deletion in the plasmid resulting in failed detection by the single target systems from Abbott and Roche used at that time, whereas the third system used, from Becton Dickinson (BD), detects nvCT. The proportion of nvCT was initially up to 65% in counties using Abbott/Roche systems. This study analysed the proportion of nvCT from 2007 to 2015 in four selected counties and its impact on chlamydia-associated complications.

Methods

C. trachomatis-positive specimens collected from 2007 to 2015 were analysed by a specific PCR to identify nvCT cases. Genotyping was performed by multilocus sequence typing (MLST) and ompA sequencing. Ectopic pregnancy and pelvic inflammatory disease records were extracted from the national registers.

Results

In total, 5101 C. trachomatis-positive samples were analysed. The nvCT proportion significantly decreased in the two counties using Roche systems, from 56% in 2007 to 6.5% in 2015 (p<0.001). In the two counties using BD systems, a decrease was also seen, from 19% in 2007 to 5.2% in 2015 (p<0.001). Fifteen nvCT cases from 2015 and 102 cases from 2006 to 2009 had identical MLST profiles. Counties using Roche/Abbott systems showed higher mean rates of ectopic pregnancy and pelvic inflammatory disease compared with counties using BD systems.

Conclusions

The nvCT proportion has decreased in all counties and converged to a low prevalence irrespective of previous rates. Genotyping showed that nvCT is clonal and genetically stable. Failing detection only marginally affected complication rates.
Predictors of mortality in Middle East respiratory syndrome (MERS)
Authors: Hong Ki-Ho , Choi Jae-Phil , Hong Seon-Hui , Lee Jeewon , Kwon Ji-Soo , Kim Sun-Mi , Park Se Yoon , Rhee Ji-Young , Kim Baek-Nam , Choi Hee Jung , Shin Eui-Cheol , Pai Hyunjoo , Park Su-Hyung , Kim Sung-Han
Published: July 2017

We evaluated the clinical characteristics, cytokine/chemokine concentrations, viral shedding and antibody kinetics in 30 patients with Middle East respiratory syndrome (MERS), including 6 non-survivors admitted to 3 MERS-designated hospitals. Old age, low albumin, altered mentality and high pneumonia severity index score at admission were risk factors for mortality. In addition, severe signs of inflammation at initial presentation (at hospital days 1-4), such as high inducible protein-10 (p=0.0013), monocyte chemoattractant protein-1 (p=0.0007) and interleukin 6 (p=0.0007) concentrations, and poor viral control (high viral load at hospital days 5–10, p<0.001) without adequate antibody titres (low antibody titre at hospital days 11–16, p=0.07) during the course of disease, were associated with mortality.
Phylogenomics reveals a complex evolutionary history of lobed-leaf white oaks in western North America
Authors: McVay John D. , Hauser Duncan , Hipp Andrew L. , Manos Paul S.
Published: July 2017

Species within the genus Quercus (oak) hybridize in complex patterns that have yet to be fully explored with phylogenomic data. Analyses to date have recovered reasonable divergent patterns, suggesting that the impact of introgression may not always be obvious in inferred oak phylogenies. We explore this phenomenon using RADseq data for 136 samples representing 54 oak species by conducting phylogenetic analyses designed to distinguish signals of lineage diversification and hybridization, focusing on the lobed-leaf species Quercus gambelii, Q. lobata, and Q. garryana in the context of a broad sampling of allied white oaks (Quercus section Quercus), and particularly the midwestern Q. macrocarpa. We demonstrate that historical introgressive hybridization between once sympatric species affects phylogeny estimation. Historical range expansion during periods of favorable climate likely explains our observations; analyses support genetic exchange between ancestral populations of Q. gambelii and Q. macrocarpa. We conclude that the genomic consequences of introgression caused the attraction of distant lineages in phylogenetic tree space, and that introgressive and divergent signals can be disentangled to produce a robust estimate of the phylogenetic history of the species.
Toward construction of a consistent field theory with Poincaré covariance in terms of step-function-type basis functions for gauge fields
Authors: Fukushima Kimichika , Sato Hikaru
Published: July 2017

This paper is a review by the authors concerning the construction of a Poincaré covariant (owing to space–time continuum) field-theoretic formalism in terms of step-function-type basis functions without ultraviolet divergences. This formalism analytically derives confinement/deconfinement, mass-gap and Regge trajectory for non-Abelian gauge fields, and gives solutions for self-interacting scalar fields. Fields propagate in space–time continuum and fields with finite degrees of freedom toward continuum limit have no ultraviolet divergence. Basis functions defined in a parameter space–time are mapped to real space–time. The authors derive a new solution comprised of classical fields as vacuum and quantum fluctuations, leading to the linear potential between the particle and antiparticle from the Wilson loop. The Polyakov line gives finite binding energies and reveals the deconfining property at high temperatures. The quantum action yields positive mass from the classical fields and quantum fluctuations produce the Coulomb potential. Pure Yang–Mills fields show the same mass-gap owing to the particle–antiparticle pair creation. The Dirac equation under linear potential is analytically solved in this formalism, reproducing the principal properties of Regge trajectories at a quantum level. Further outlook mentions a possibility of the difference between conventional continuum and present wave functions responsible for the cosmological constant.
Fast Frequency-Domain Algorithm for Estimating the Dynamic Wind-Induced Response of Large-Span Roofs Based on Cauchy’s Residue Theorem
Authors: Su Ning , Cao Zhenggang , Wu Yue
Published: July 2017

Wind-induced response analysis is an important process in the design of large-span roofs. Conventional time-domain methods are computationally more expensive than frequency-domain algorithms; however, the latter are not as accurate because of the ill-treatment of the modal coupling effects. This paper revisited the derivations of the frequency-domain algorithm and proposed a fast algorithm for estimating the dynamic wind-induced response considering duly the modal coupling effects. With the wind load cross-spectra modeled by rational functions, closed-form solutions to the frequency-domain integrals can be calculated by Cauchy’s residue theorem, rather than by numerical integration, thereby reducing the truncation errors and enhancing the efficiency of computation. The algorithm is applied to the analysis of a grandstand roof and a spherical dome. Through comparison with time domain analyses results, the algorithm is proved to be reliable. A criterion of the coupling modal combination was suggested based on the cumulative modal contribution rate of over 70%.
Buckling of Graphene Platelet Reinforced Composite Cylindrical Shell with Cutout
Authors: Wang Yu , Feng Chuang , Zhao Zhan , Yang Jie
Published: July 2017

This paper investigates the buckling behavior of graphene platelets (GPL) reinforced composite cylindrical shells with cutouts via finite element method (FEM) simulation. Young’s modulus of the composites is determined by the modified Halpin–Tsai micromechanics model while the mass density and Poisson’s ratio of the composites are approximated by the rule of mixture. Comprehensive parametric study is conducted to investigate the effects of the weight fraction and the shape of GPL fillers, the geometry of the shell and the position and orientation of the cutout on the buckling behaviors of the cylindrical structures. The results demonstrate that the addition of GPLs can significantly increase the load bearing capacity of the cylindrical shells. Larger sized GPLs with fewer single graphene layers are favorable reinforcing fillers in enhancing the buckling performances of the structures. The buckling load is sensitive to the location of the cutout with larger aspect ratio. Moreover, the orientation of the cutout is found to have significant effects on the buckling load when the orientation angle [Formula: see text] is falling within the ranges – [Formula: see text]/2 [Formula: see text] – [Formula: see text]/4 and [Formula: see text]/4 [Formula: see text]/2.
OntoDTA: Ontology-Guided Decision Tree Assistance
Authors: Benali Khaled , Rahal Sidi Ahmed
Published: July 2017

The effective application of a Decision Tree (DT) process is beset with many difficult and technical decisions about the choice of algorithms, parameters, evaluation, etc. Therefore, we propose assistance by using ontologies for addressing the above-mentioned challenges that face the non-specialist DT miner (person). Ontologies have been used in various research areas such as computer science, including data mining tools. In this paper, we propose the realisation of a domain ontology for DT OntoDTA to empower the non-specialist DT miner throughout the key phases of the DT process. OntoDTA ontology contains the knowledge of DT and provides a common terminology that can be shared and processed by DT miners.
Study of particles from the Kafang copper deposit in Gejiu city, Yunnan
Authors: Li Dewei , Cao Jianjin , Ke Hongling , Liu Chang , Wei Xiaojun
Published: July 2017

Samples were collected from the mining area, limestone area, and prospective area proximal to the Kafang copper deposit. The particles were analysed by the transmission electron microscopy (TEM) and the soil samples in the mining area were analysed by the X-ray diffraction (XRD). In the mining area and prospective area, metallic elements in the particles are Cu, Pb, Zn, As, Hg, and Co. Particles collected in the limestone area contain few metallic elements that exist in the ore minerals. Based on analysis of these particles, the following conclusions are made: (1) sizes of the collected particles are 20 – 300 nm; (2) the composition of the soil samples and the particulate samples are significantly different because particles in the particulate samples reflect an origin from geogas; (3) geogas particles have different elemental contents because of different physicochemical properties of the elements and physicochemical environment that the geogas particles experienced; and (4) the Kafang stratiform copper deposit is a granite-related hydrothermal deposit. Metallic elements in geogas particles occur in the ore minerals and have similar compositions to those of geogas particles in the sampling sites. The geogas particles most likely originate from concealed deposits.

Supplementary material:
EDS spectra of particles and coordinates of sampling points in the mining area, limestone area, and prospecting area are available at:
https://doi.org/10.6084/m9.figshare.c.3820687
Proof-Carrying Hardware via Inductive Invariants
Authors: Isenberg Tobias , Platzner Marco , Wehrheim Heike , Wiersema Tobias
Published: July 2017

Proof-carrying hardware (PCH) is a principle for achieving safety for dynamically reconfigurable hardware systems. The producer of a hardware module spends huge effort when creating a proof for a safety policy. The proof is then transferred as a certificate together with the configuration bitstream to the consumer of the hardware module, who can quickly verify the given proof. Previous work utilized SAT solvers and resolution traces to set up a PCH technology and corresponding tool flows. In this article, we present a novel technology for PCH based on inductive invariants. For sequential circuits, our approach is fundamentally stronger than the previous SAT-based one since we avoid the limitations of bounded unrolling. We contrast our technology to existing ones and show that it fits into previously proposed tool flows. We conduct experiments with four categories of benchmark circuits and report consumer and producer runtime and peak memory consumption, as well as the size of the certificates and the distribution of the workload between producer and consumer. Experiments clearly show that our new induction-based technology is superior for sequential circuits, whereas the previous SAT-based technology is the better choice for combinational circuits.
Vagally mediated effects of brain stem dopamine on gastric tone and phasic contractions of the rat
Authors: Anselmi L. , Toti L. , Bove C. , Travagli R. A.
Published: July 2017

Dopamine (DA)-containing fibers and neurons are embedded within the brain stem dorsal vagal complex (DVC); we have shown previously that DA modulates the membrane properties of neurons of the dorsal motor nucleus of the vagus (DMV) via DA1 and DA2 receptors. The vagally dependent modulation of gastric tone and phasic contractions, i.e., motility, by DA, however, has not been characterized. With the use of microinjections of DA in the DVC while recording gastric tone and motility, the aims of the present study were 1) assess the gastric effects of brain stem DA application, 2) identify the DA receptor subtype, and, 3) identify the postganglionic pathway(s) activated. Dopamine microinjection in the DVC decreased gastric tone and motility in both corpus and antrum in 29 of 34 rats, and the effects were abolished by ipsilateral vagotomy and fourth ventricular treatment with the selective DA2 receptor antagonist L741,626 but not by application of the selective DA1 receptor antagonist SCH 23390. Systemic administration of the cholinergic antagonist atropine attenuated the inhibition of corpus and antrum tone in response to DA microinjection in the DVC. Conversely, systemic administration of the nitric oxide synthase inhibitor nitro-l-arginine methyl ester did not alter the DA-induced decrease in gastric tone and motility. Our data provide evidence of a dopaminergic modulation of a brain stem vagal neurocircuit that controls gastric tone and motility.

NEW & NOTEWORTHY Dopamine administration in the brain stem decreases gastric tone and phasic contractions. The gastric effects of dopamine are mediated via dopamine 2 receptors on neurons of the dorsal motor nucleus of the vagus. The inhibitory effects of dopamine are mediated via inhibition of the postganglionic cholinergic pathway.
Developing better mouse models to study cisplatin-induced kidney injury
Authors: Sharp Cierra N. , Siskind Leah J.
Published: July 2017

Cisplatin is a potent chemotherapeutic used for the treatment of many types of cancer. However, its dose-limiting side effect is nephrotoxicity leading to acute kidney injury (AKI). Patients who develop AKI have an increased risk of mortality and are more likely to develop chronic kidney disease (CKD). Unfortunately, there are no therapeutic interventions for the treatment of AKI. It has been suggested that the lack of therapies is due in part to the fact that the established mouse model used to study cisplatin-induced AKI does not recapitulate the cisplatin dosing regimen patients receive. In recent years, work has been done to develop more clinically relevant models of cisplatin-induced kidney injury, with much work focusing on incorporation of multiple low doses of cisplatin administered over a period of weeks. These models can be used to recapitulate the development of CKD after AKI and, by doing so, increase the likelihood of identifying novel therapeutic targets for the treatment of cisplatin-induced kidney injury.
Acute volume loading exacerbates direct ventricular interaction in a model of COPD
Authors: Cheyne William S. , Williams Alexandra M. , Harper Megan I. , Eves Neil D.
Published: July 2017

Volume loading increases left ventricular (LV) stroke volume (LVSV) through series interaction, but may paradoxically reduce LVSV in the presence of large increases in right ventricular (RV) afterload because of direct ventricular interaction (DVI). RV afterload is often increased in chronic obstructive pulmonary disease (COPD) as a result of pathological changes to respiratory mechanics, namely increased negative intrathoracic pressure (nITP), dynamic lung hyperinflation (DH), and increased pulmonary vascular resistance (PVR). These hallmarks of COPD negatively impact LV hemodynamics in normovolemia. However, it is unknown how these heart-lung interactions are impacted by acute volume loading. Twenty healthy subjects (23 ± 2 yr) completed the study protocol, involving acute volume loading via 20° head-down tilt (HDT) in isolation and with 1) inspiratory resistance of −20 cmH2O (HDT+nITP) and 2) nITP, expiratory resistance to induce DH and hypoxic-mediated increases in PVR (HDT+COPD model). LV volumes and geometry were assessed using triplane echocardiography. HDT significantly increased LVSV by 10 ± 10% through an 8 ± 6% increase in LV end-diastolic volume (LVEDV). HDT+nITP paradoxically decreased LVSV by 11 ± 12% and LVEDV by 6 ± 9% from supine baseline, or −14 ± 10% LVSV and −15 ± 13% LVEDV from HDT ( P < 0.001). HDT+COPD model decreased LVSV (21 ± 10% and 28 ± 11%) and LVEDV (16 ± 10% and 22 ± 10%) from both supine and HDT, respectively ( P < 0.001). Under all conditions, significant septal flattening (increased radius of septal curvature) occurred, indicating DVI. Thus, when RV afterload is increased and/or an external constraint to ventricular filling exists, acute volume loading appears to paradoxically reduce LVSV. These findings have important implications for understanding how volume status impacts cardiopulmonary interactions in COPD.

NEW & NOTEWORTHY Volume loading may exacerbate adverse cardiopulmonary interaction in COPD; however, the mechanisms remain unclear. We found that when negative intrathoracic pressure is increased, acute volume loading paradoxically reduces stroke volume. This reduction in stroke volume is considerably greater in a model of COPD, owing to the effects of lung hyperinflation.
A Multiscale Model for the World’s First Parasitic Disease Targeted for Eradication: Guinea Worm Disease
Authors: Netshikweta Rendani , Garira Winston
Published: July 2017

Guinea worm disease (GWD) is both a neglected tropical disease and an environmentally driven infectious disease. Environmentally driven infectious diseases remain one of the biggest health threats for human welfare in developing countries and the threat is increased by the looming danger of climate change. In this paper we present a multiscale model of GWD that integrates the within-host scale and the between-host scale. The model is used to concurrently examine the interactions between the three organisms that are implicated in natural cases of GWD transmission, the copepod vector, the human host, and the protozoan worm parasite (Dracunculus medinensis), and identify their epidemiological roles. The results of the study (through sensitivity analysis of R0) show that the most efficient elimination strategy for GWD at between-host scale is to give highest priority to copepod vector control by killing the copepods in drinking water (the intermediate host) by applying chemical treatments (e.g., temephos, an organophosphate). This strategy should be complemented by health education to ensure that greater numbers of individuals and communities adopt behavioural practices such as voluntary reporting of GWD cases, prevention of GWD patients from entering drinking water bodies, regular use of water from safe water sources, and, in the absence of such water sources, filtering or boiling water before drinking. Taking into account the fact that there is no drug or vaccine for GWD (interventions which operate at within-host scale), the results of our study show that the development of a drug that kills female worms at within-host scale would have the highest impact at this scale domain with possible population level benefits that include prevention of morbidity and prevention of transmission.
An Improved Extrapolation Scheme for Truncated CT Data Using 2D Fourier-Based Helgason-Ludwig Consistency Conditions
Authors: Xia Yan , Berger Martin , Bauer Sebastian , Hu Shiyang , Aichert Andre , Maier Andreas
Published: July 2017

We improve data extrapolation for truncated computed tomography (CT) projections by using Helgason-Ludwig (HL) consistency conditions that mathematically describe the overlap of information between projections. First, we theoretically derive a 2D Fourier representation of the HL consistency conditions from their original formulation (projection moment theorem), for both parallel-beam and fan-beam imaging geometry. The derivation result indicates that there is a zero energy region forming a double-wedge shape in 2D Fourier domain. This observation is also referred to as the Fourier property of a sinogram in the previous literature. The major benefit of this representation is that the consistency conditions can be efficiently evaluated via 2D fast Fourier transform (FFT). Then, we suggest a method that extrapolates the truncated projections with data from a uniform ellipse of which the parameters are determined by optimizing these consistency conditions. The forward projection of the optimized ellipse can be used to complete the truncation data. The proposed algorithm is evaluated using simulated data and reprojections of clinical data. Results show that the root mean square error (RMSE) is reduced substantially, compared to a state-of-the-art extrapolation method.
Oral Administration of the Endocannabinoid Anandamide during Lactation: Effects on Hypothalamic Cannabinoid Type 1 Receptor and Food Intake in Adult Mice
Authors: Aguirre Carolina , Castillo Valeska , Llanos Miguel
Published: July 2017

We have previously shown that administration of the endocannabinoid anandamide (AEA) during lactation leads to overweight, increased body fat accumulation, and insulin resistance in adult mice. This study was designed to elucidate if these effects are due to increased food intake, stimulated by an augmented abundance and binding ability of the hypothalamic cannabinoid type 1 receptor (CB1R). With this aim, male mice pups were treated with a daily oral dose of AEA during lactation. Adult mice were also treated with a single oral dose of AEA, to evaluate acute food intake during 4 h. At 21 and 160 days, CB1R protein abundance was calculated by western blot analysis. Capacity of hypothalamic membranes to specifically bind the radioligand3[H]-CP55.940 was also measured. Western blots showed a 72% increase in CB1R abundance in AEA-treated 21-day-old mice, without differences in adult mice. Additionally, specific binding of3[H]-CP55.940 to hypothalamic membranes from adult mice was significantly lower in those mice treated with AEA during lactation. Moreover, AEA did not stimulate acute food intake in both, AEA-treated and control mice. Results suggest that metabolic alterations found in adult mice because of AEA treatment during lactation are not associated with hypothalamic CB1R.
Expression of Telomere Repeat Binding Factor 1 and TRF2 in Prostate Cancer and Correlation with Clinical Parameters
Authors: Chen Wei , Wang Yong , Li Fei , Lin Wei , Liang Yong , Ma Zhiwei
Published: July 2017

Objective. The objective of this study was to investigate the expression of telomere repeat binding factor 1 (TRF1) and TRF2 in prostate cancer and their relationships with clinicopathological features. Methods. In total 50 prostate cancer tissues and paired benign prostate hyperplasia tissues were analyzed. The telomere-binding proteins TRF1 and TRF2 were measured using immunohistochemical method. Correlation analyses were used to evaluate the association between immunohistochemical score and clinical parameters. Results. The expression of TRF1 was significantly higher in prostate cancer tissue than in benign prostate hyperplasia tissue (χ2 = 62.69, P<0.01). Elevated levels of TRF2 were observed in both prostate cancer and benign prostate hyperplasia tissue (χ2 = 1.13, P=0.76). TRF1 expression was significantly positively correlated with surgical capsular invasion (Spearman’s r=0.43, P=0.002), seminal vesicle invasion (Spearman’s r=0.35, P=0.01), lymph nodes metastases (Spearman’s r=0.41, P=0.003), total prostate specific antigen (r=0.61, P<0.05), and Gleason score (r=0.47, P=0.01). However, there were no significant statistical differences between prostate volume (r=0.06, P=0.75) and age (r=0.14, P=0.09). Conclusion. Both TRF1 and TRF2 were overexpressed in prostate cancer. There was no specificity of TRF2 in prostate cancer, while TRF1 may be associated with prostate cancer progression.
Identification of New Tumor Suppressor Genes in Triple-Negative Breast Cancer
Authors: Rangel Roberto , Guzman-Rojas Liliana , Kodama Takahiro , Kodama Michiko , Newberg Justin Y. , Copeland Neal G. , Jenkins Nancy A.
Published: July 2017

Although genomic sequencing has provided a better understating of the genetic landmarks in triple-negative breast cancer (TNBC), functional validation of candidate cancer genes (CCG) remains unsolved. In this study, we used a transposon mutagenesis strategy based on a two-step sleeping beauty (SB) forward genetic screen to identify and validate new tumor suppressors (TS) in this disease. We generated 120 siRNAs targeting 40 SB-identified candidate breast cancer TS genes and used them to downregulate expression of these genes in four human TNBC cell lines. Among CCG, whose SB-mediated genetic mutation resulted in increased cellular proliferation in all cell lines tested, the genes ADNP, AP2B1, TOMM70A, and ZNF326 showed TS activity in tumor xenograft studies. Subsequent studies showed that ZNF326 regulated expression of multiple epithelial–mesenchymal transition and cancer stem cell (CSC) pathway genes. It also modulated expression of TS genes involved in the regulation of migration and cellular invasion and was a direct transcriptional activator of genes that regulate CSC self-renewal. ZNF326 expression associated with TNBC patient survival, with ZNF326 protein levels showing a marked reduction in TNBC. Our validation of several new TS genes in TNBC demonstrate the utility of two-step forward genetic screens in mice and offer an invaluable tool to identify novel candidate therapeutic pathways and targets. Cancer Res; 77(15); 4089–101. ©2017 AACR.
Germline Mutations in Cancer Susceptibility Genes in a Large Series of Unselected Breast Cancer Patients
Authors: Sun Jie , Meng Hua , Yao Lu , Lv Meng , Bai Jian , Zhang Jianguang , Wang Lientu , Ouyang Tao , Li Jinfeng , Wang Tianfeng , Fan Zhaoqing , Fan Tie , Lin Benyao , Xie Yuntao
Published: July 2017

Purpose: The prevalence of mutations in cancer susceptibility genes such as BRCA1 and BRCA2 and other cancer susceptibility genes and their clinical relevance are largely unknown among a large series of unselected breast cancer patients in the Chinese population.

Experimental Design: A total of 8,085 consecutive unselected Chinese breast cancer patients were enrolled. Germline mutations in 46 cancer susceptibility genes were detected using a 62-gene panel.

Results: Pathogenic mutations were identified in 9.2% of patients among the 8,085 unselected breast cancer patients. Of these, 5.3% of patients carried a BRCA1 or BRCA2 mutation (1.8% in BRCA1 and 3.5% in BRCA2), 2.9% carried other breast cancer susceptibility genes (BOCG) and 1.0% carried another cancer susceptibility genes. Triple-negative breast cancers had the highest prevalence of BRCA1/2 mutations (11.2%) and other BOCG mutations (3.8%) among the four molecular subgroups, whereas ER−/PR−HER2+ breast cancers had the lowest mutations in BRCA1/2 (1.8%) and BOCG (1.6%). In addition, BRCA1 mutation carriers had a significant worse disease-free survival [unadjusted hazard ratio (HR) 1.60; 95% confidence interval (CI) 1.10–2.34; P = 0.014] and disease-specific survival (unadjusted HR 1.96; 95% CI, 1.03–3.65; P = 0.040) than did non-carriers, whereas no significant difference in survival was found between BRCA2 mutation carriers and non-carriers.

Conclusions: 9.2% of breast cancer patients carry a pathogenic mutation in cancer susceptibility genes in this large unselected series. Triple-negative breast cancers have the highest prevalence of mutations in BRCA1 /2 and other breast cancer susceptibility genes among the four molecular subgroups, whereas ER−/PR−HER2+ breast cancers had the lowest mutations in these genes. Clin Cancer Res; 23(20); 6113–9. ©2017 AACR.
BRAF Inhibitors Amplify the Proapoptotic Activity of MEK Inhibitors by Inducing ER Stress in NRAS-Mutant Melanoma
Authors: Niessner Heike , Sinnberg Tobias , Kosnopfel Corinna , Smalley Keiran S.M. , Beck Daniela , Praetorius Christian , Mai Marion , Beissert Stefan , Kulms Dagmar , Schaller Martin , Garbe Claus , Flaherty Keith T. , Westphal Dana , Wanke Ines , Meier Friedegund
Published: July 2017

Purpose: NRAS mutations in malignant melanoma are associated with aggressive disease requiring rapid antitumor intervention, but there is no approved targeted therapy for this subset of patients. In clinical trials, the MEK inhibitor (MEKi) binimetinib displayed modest antitumor activity, making combinations a requisite. In a previous study, the BRAF inhibitor (BRAFi) vemurafenib was shown to induce endoplasmic reticulum (ER) stress that together with inhibition of the RAF–MEK–ERK (MAPK) pathway amplified its proapoptotic activity in BRAF-mutant melanoma. The present study investigated whether this effect might extent to NRAS-mutant melanoma, in which MAPK activation would be expected.

Experimental Design and Results: BRAFi increased pERK, but also significantly increased growth inhibition and apoptosis induced by the MEKi in monolayer, spheroids, organotypic, and patient-derived tissue slice cultures of NRAS-mutant melanoma. BRAFi such as encorafenib induced an ER stress response via the PERK pathway, as detected by phosphorylation of eIF2α and upregulation of the ER stress–related factors ATF4, CHOP, and NUPR1 and the proapoptotic protein PUMA. MEKi such as binimetinib induced the expression of the proapoptotic protein BIM and activation of the mitochondrial pathway of apoptosis, the latter of which was enhanced by combination with encorafenib. The increased apoptotic rates caused by the combination treatment were significantly reduced through siRNA knockdown of ATF4 and BIM, confirming its critical roles in this process.

Conclusions: The data presented herein encourage further advanced in vivo and clinical studies to evaluate MEKi in combination with ER stress inducing BRAFi as a strategy to treat rapidly progressing NRAS-mutant melanoma. Clin Cancer Res; 23(20); 6203–14. ©2017 AACR.
The c.1085A>G Genetic Variant of CSF1R Gene Regulates Tumor Immunity by Altering the Proliferation, Polarization, and Function of Macrophages
Authors: Yeh Yu-Min , Hsu Shan-Ju , Lin Peng-Chan , Hsu Keng-Fu , Wu Pei-Ying , Su Wu-Chou , Chang Jang-Yang , Shen Meng-Ru
Published: July 2017

Purpose: Targeting tumor-associated macrophages with colony-stimulating factor 1 receptor (CSF-1R) inhibition reveals a strategy for cancer therapy. Here, we studied the impact of CSF1R germline genetic variant on CSF-1R signaling and the susceptibility to CSF-1R inhibitors.

Experimental designs: CSF1R germline genetic variants were studied in 140 cancer patients. CSF-1R phosphorylation, endocytosis, and macrophage polarization were measured as the response to CSF-1 stimulation. Tumor-associated macrophages in surgical specimens and sensitivity to CSF-1R inhibitors were used to determine macrophage function.

Results: A CSF1R c.1085A>G genetic variant causing the change of histidine to arginine in the domain of receptor dimerization was identified as a high allele frequency in Eastern Asian population. Cancer patients with this variant allele had less M2-like tumor-associated macrophages accompanied by low VEGF expression in tumor tissues. Importantly, CSF1R genetic variant was significantly associated with disease-free survival in colorectal, endometrial, and ovarian cancer. In terms of differentiation, macrophages with CSF1R c.1085A>G genetic variant displayed a refractory response to CSF-1 stimulation and macrophage survival was sensitive to CSF-1R inhibitors with IC50 of 0.1 to 1 nmol/L range. On contrast, CSF-1 induced a prominent phosphorylation and rapid endocytosis of CSF-1R, leading to an M2-like dominant polarization in macrophages with CSF1R c.1085 genotype A_A, in which CSF-1R inhibitors of PLX3397, BLZ945, and GW2580 inhibited macrophage survival with IC50 of 10 to 100 nmol/L range.

Conclusions: The CSF1R c.1085A>G genetic variant regulates tumor immunity by altering the polarization and function of macrophages. This genetic variant confers the sensitivity to CSF-1R inhibitors, implying as a biomarker in targeting CSF-1R signaling for cancer treatment. Clin Cancer Res; 23(20); 6021–30. ©2017 AACR.
MICA-Expressing Monocytes Enhance Natural Killer Cell Fc Receptor-Mediated Antitumor Functions
Authors: Campbell Amanda R. , Duggan Megan C. , Suarez-Kelly Lorena P. , Bhave Neela , Opheim Kallan S. , McMichael Elizabeth L. , Trikha Prashant , Parihar Robin , Luedke Eric , Lewis Adrian , Yung Bryant , Lee Robert , Raulet David , Tridandapani Susheela , Groh Veronika , Yu Lianbo , Yildiz Vedat , Byrd John C. , Caligiuri Michael A. , Carson William E.
Published: July 2017

Natural killer (NK) cells are large granular lymphocytes that promote the antitumor response via communication with other cell types in the tumor microenvironment. Previously, we have shown that NK cells secrete a profile of immune stimulatory factors (e.g., IFNγ, MIP-1α, and TNFα) in response to dual stimulation with the combination of antibody (Ab)-coated tumor cells and cytokines, such as IL12. We now demonstrate that this response is enhanced in the presence of autologous monocytes. Monocyte enhancement of NK cell activity was dependent on cell-to-cell contact as determined by a Transwell assay. It was hypothesized that NK cell effector functions against Ab-coated tumor cells were enhanced via binding of MICA on monocytes to NK cell NKG2D receptors. Strategies to block MICA–NKG2D interactions resulted in reductions in IFNγ production. Depletion of monocytes in vivo resulted in decreased IFNγ production by murine NK cells upon exposure to Ab-coated tumor cells. In mice receiving trastuzumab and IL12 therapy, monocyte depletion resulted in significantly greater tumor growth in comparison to mock-depleted controls (P < 0.05). These data suggest that NK cell–monocyte interactions enhance NK cell antitumor activity in the setting of monoclonal Ab therapy for cancer. Cancer Immunol Res; 5(9); 778–89. ©2017 AACR.