bluuebunny/crossref_metadata_2025 huggingface dataset:
The challenges of detecting subtle population structure and its importance for the conservation of emperor penguins
Authors: Younger Jane L. , Clucas Gemma V. , Kao Damian , Rogers Alex D. , Gharbi Karim , Hart Tom , Miller Karen J.
Published: May 2017

Understanding the boundaries of breeding populations is of great importance for conservation efforts and estimates of extinction risk for threatened species. However, determining these boundaries can be difficult when population structure is subtle. Emperor penguins are highly reliant on sea ice, and some populations may be in jeopardy as climate change alters sea‐ice extent and quality. An understanding of emperor penguin population structure is therefore urgently needed. Two previous studies have differed in their conclusions, particularly whether the Ross Sea, a major stronghold for the species, is isolated or not. We assessed emperor penguin population structure using 4,596 genome‐wide single nucleotide polymorphisms (SNPs), characterized in 110 individuals (10–16 per colony) from eight colonies around Antarctica. In contrast to a previous conclusion that emperor penguins are panmictic around the entire continent, we find that emperor penguins comprise at least four metapopulations, and that the Ross Sea is clearly a distinct metapopulation. Using larger sample sizes and a thorough assessment of the limitations of different analytical methods, we have shown that population structure within emperor penguins does exist and argue that its recognition is vital for the effective conservation of the species. We discuss the many difficulties that molecular ecologists and managers face in the detection and interpretation of subtle population structure using large SNP data sets, and argue that subtle structure should be taken into account when determining management strategies for threatened species, until accurate estimates of demographic connectivity among populations can be made.
Sex‐specific graphs: Relating group‐specific topology to demographic and landscape data
Authors: Bertrand Philip , Bowman Jeff , Dyer Rodney J. , Manseau Micheline , Wilson Paul J.
Published: May 2017

Sex‐specific genetic structure is a commonly observed pattern among vertebrate species. Facing differential selective pressures, individuals may adopt sex‐specific life history traits that ultimately shape genetic variation among populations. Although differential dispersal dynamics are commonly detected in the literature, few studies have used genetic structure to investigate sex‐specific functional connectivity. The recent use of graph theoretic approaches in landscape genetics has demonstrated network capacities to describe complex system behaviours where network topology represents genetic interaction among subunits. Here, we partition the overall genetic structure into sex‐specific graphs, revealing different male and female dispersal dynamics of a fisher (Pekania [Martes] pennanti) metapopulation in southern Ontario. Our analyses based on network topologies supported the hypothesis of male‐biased dispersal. Furthermore, we demonstrated that the effect of the landscape, identified at the population level, could be partitioned among sex‐specific strata. We found that female connectivity was negatively correlated with snow depth, whereas connectivity among males was not. Our findings underscore the potential of conducting sex‐specific analysis by identifying landscape elements or configuration that differentially promotes or impedes functional connectivity between sexes, revealing processes that may otherwise remain cryptic. We propose that the sex‐specific graph approach would be applicable to other vagile species where differential sex‐specific processes are expected to occur.
Diagnostic anoctamin‐5 protein defect in patients with ANO5‐mutated muscular dystrophy
Authors: Vihola A. , Luque H. , Savarese M. , Penttilä S. , Lindfors M. , Leturcq F. , Eymard B. , Tasca G. , Brais B. , Conte T. , Charton K. , Richard I. , Udd B.
Published: May 2017

Aims

Previously, detection of ANO5 protein has been complicated by unspecific antibodies, most of which have not identified the correct protein. The aims of the study were to specify ANO5 protein expression in human skeletal muscle, and to investigate if the ANO5 protein levels are affected by different ANO5 mutations in anoctaminopathy patients.

Methods

Four different antibodies were tested for ANO5 specificity. A sample preparation method compatible with membrane proteins, combined with tissue fractionation was used to determine ANO5 expression in cell cultures expressing ANO5, in normal muscles and eight patient biopsies with six different ANO5 mutations in homozygous or compound heterozygous states, and in other dystrophies.

Results

Only one specific monoclonal N‐terminal ANO5 antibody was efficient in detecting the protein, showing that ANO5 is expressed as a single 107 kD polypeptide in human skeletal muscle. The truncating mutations c.191dupA and c.1261C>T were found to abolish ANO5 expression, whereas the studied point mutations had variable effects; however, all the ANO5 mutations resulted in clearly reduced ANO5 expression in the patient muscle membrane fraction. Attempts to detect ANO5 using immunohistochemistry were not yet successful.

Conclusions

The data presented here indicate that the ANO5 protein expression is decreased in ANO5‐mutated muscular dystrophy and that most of the non‐truncating pathogenic ANO5 mutations likely destabilize the protein and cause its degradation. The method described here allows direct analysis of human ANO5 protein, which can be used in diagnostics, for evaluating the pathogenicity of the potentially harmful ANO5 variants of uncertain significance.
Commentary: Effects of Success on Management Performance
Authors: Stanton Thomas H.
Published: May 2017

Related Content: Nicholson‐Crotty, Nicholson‐Crotty and Fernandez (PAR July/August 2017)
CT‐determined pulmonary artery to aorta ratio as a predictor of elevated pulmonary artery pressure and survival in idiopathic pulmonary fibrosis
Authors: Yagi Mitsuaki , Taniguchi Hiroyuki , Kondoh Yasuhiro , Ando Masahiko , Kimura Tomoki , Kataoka Kensuke , Furukawa Taiki , Suzuki Atsushi , Johkoh Takeshi , Hasegawa Yoshinori
Published: May 2017

ABSTRACT

Background and objective

Elevated mean pulmonary artery pressure (mPAP) is a significant prognostic indicator in idiopathic pulmonary fibrosis (IPF). It has been reported that the computed tomography‐determined ratio of the diameter of the pulmonary artery to the aorta (PA:A) is correlated with mPAP in various respiratory diseases. However, in patients with IPF, whether the PA:A can be used to predict elevated mPAP and the prognostic value of the PA:A has not been fully evaluated.

Methods

We conducted a single‐centre, observational study of 177 consecutive IPF patients who underwent right heart catheterization. We examined the association between the PA:A and mPAP in these patients, and performed a receiver operating characteristic (ROC) analysis to test the diagnostic accuracy of the PA:A in predicting mPAP > 20 mm Hg (pulmonary hypertension (PH) or borderline PH) in relation to survival.

Results

In a multivariate linear regression analysis, the PA:A, 6‐min walk distance and diffusion capacity for carbon monoxide (DLCO
) % predicted were independent explanatory variables of elevated mPAP (P < 0.0001, P = 0.009 and P = 0.002, respectively). ROC analysis revealed a PA:A > 0.9 to be optimal for predicting mPAP > 20 mmHg (area under the curve (AUC) = 0.75; 95% CI: 0.65–0.84). Patients with a PA:A > 0.9 also had a worse prognosis (P = 0.009).

Conclusion

Measurement of the PA:A is a useful and convenient method to predict elevated mPAP in IPF at initial evaluation. Moreover, a PA:A >0.9 was found to be an indicator of worse prognosis.
Will the safe‐breakers of sleep apnoea find the right combination?
Authors: Grunstein Ronald R.
Published: May 2017

See DOI: 10.1111/resp.13044
Systematic review of exercise training in colorectal cancer patients during treatment
Authors: van Rooijen S. J. , Engelen M. A. , Scheede‐Bergdahl C. , Carli F. , Roumen R. M. H. , Slooter G. D. , Schep G.
Published: May 2017

Colorectal cancer surgery results in considerable postoperative morbidity, mortality and reduced quality of life. As many patients will undergo additional (neo)adjuvant therapy, it is imperative that each individual optimize their physical function. To elucidate the potential of exercise in patient optimization, we investigated the evidence for an exercise program before and after surgical treatment in colorectal cancer patients. A systematic review was conducted according to the Cochrane Handbook for Systematic Reviews of Interventions, the guidelines of the Physical Therapy Journal and the PRISMA guidelines. No literature pertaining to exercise training during preoperative neoadjuvant treatment was found. Seven studies, investigating the effects of regular exercise during adjuvant chemotherapy for patients with colorectal cancer or a mixed population, were identified. A small effect (effect size (ES) 0.4) of endurance/interval training and strength training (ES 0.4) was found in two studies conducted in patients with colorectal and gastrointestinal cancer. In five studies that included a mixed population of cancer patients, interval training resulted in a large improvement (ES 1.5; P≤.05). Endurance training alone was found to increase both lower extremity strength and endurance capacity. The effects of strength training in the lower extremity are moderate, whereas, in the upper extremity, the increase is small. There is limited evidence available on exercise training during treatment in colorectal cancer patients. One study concluded exercise therapy may be beneficial for colorectal cancer patients during adjuvant treatment. The possible advantages of training during neoadjuvant treatment may be explored by prehabilitation trials.
Locating the External Source of Enforceability: Alliances, Bilateral Investment Treaties, and Foreign Direct Investment
Authors: Wang Zhiyuan , Youn Hyunjin
Published: May 2017

Objective

We theorize that alliances play a role as an external source of enforceability of bilateral investment treaties (BITs), and they do this by limiting the policy opportunism in foreign direct investment (FDI) host countries. We attempt to demonstrate that being party to common BITs and common alliances substantially raises the accumulation of FDI.

Methods

Using econometric techniques including panel‐corrected standard error, fixed effect, and dynamic pooled models, we analyze panel data on dyadic FDI stock from 1978 to 2003.

Results

We find that BITs increase FDI stock and that alliances magnify this positive effect of BITs by a substantial margin.

Conclusion

Being embedded in alliances strengthens the impact of BITs on FDI. This study contributes to the extant literature on the efficacy of BITs in particular and that on international institutions in general.
A support theorem for the Hitchin fibration: the case of
Authors: de Cataldo Mark Andrea
Published: May 2017

We prove that the direct image complex for the$D$-twisted$\text{SL}_{n}$Hitchin fibration is determined by its restriction to the elliptic locus, where the spectral curves are integral. The analogous result for$\text{GL}_{n}$is due to Chaudouard and Laumon. Along the way, we prove that the Tate module of the relative Prym group scheme is polarizable, and we also prove$\unicode[STIX]{x1D6FF}$-regularity results for some auxiliary weak abelian fibrations.
A Variant PfCRT Isoform Can Contribute to
Plasmodium falciparum
Resistance to the First-Line Partner Drug Piperaquine
Authors: Dhingra Satish K. , Redhi Devasha , Combrinck Jill M. , Yeo Tomas , Okombo John , Henrich Philipp P. , Cowell Annie N. , Gupta Purva , Stegman Matthew L. , Hoke Jonathan M. , Cooper Roland A. , Winzeler Elizabeth , Mok Sachel , Egan Timothy J. , Fidock David A.
Published: May 2017

Current efforts to reduce the global burden of malaria are threatened by the rapid spread throughout Asia of
Plasmodium falciparum
resistance to artemisinin-based combination therapies, which includes increasing rates of clinical failure with dihydroartemisinin plus piperaquine (PPQ) in Cambodia. Using zinc finger nuclease-based gene editing, we report that addition of the C101F mutation to the chloroquine (CQ) resistance-conferring PfCRT Dd2 isoform common to Asia can confer PPQ resistance to cultured parasites. Resistance was demonstrated as significantly higher PPQ concentrations causing 90% inhibition of parasite growth (IC
90
) or 50% parasite killing (50% lethal dose [LD
50
]). This mutation also reversed Dd2-mediated CQ resistance, sensitized parasites to amodiaquine, quinine, and artemisinin, and conferred amantadine and blasticidin resistance. Using heme fractionation assays, we demonstrate that PPQ causes a buildup of reactive free heme and inhibits the formation of chemically inert hemozoin crystals. Our data evoke inhibition of heme detoxification in the parasite’s acidic digestive vacuole as the primary mode of both the
bis
-aminoquinoline PPQ and the related 4-aminoquinoline CQ. Both drugs also inhibit hemoglobin proteolysis at elevated concentrations, suggesting an additional mode of action. Isogenic lines differing in their
pfmdr1
copy number showed equivalent PPQ susceptibilities. We propose that mutations in PfCRT could contribute to a multifactorial basis of PPQ resistance in field isolates.

IMPORTANCE
The global agenda to eliminate malaria depends on the continued success of artemisinin-based combination therapies (ACTs), which target the asexual blood stages of the intracellular parasite
Plasmodium
. Partial resistance to artemisinin, however, is now established in Southeast Asia, exposing the partner drugs to increased selective pressure.
Plasmodium falciparum
resistance to the first-line partner piperaquine (PPQ) is now spreading rapidly in Cambodia, resulting in clinical treatment failures. Here, we report that a variant form of the
Plasmodium falciparum
chloroquine resistance transporter, harboring a C101F mutation edited into the chloroquine (CQ)-resistant Dd2 isoform prevalent in Asia, can confer PPQ resistance in cultured parasites. This was accompanied by a loss of CQ resistance. Biochemical assays showed that PPQ, like CQ, inhibits the detoxification of reactive heme that is formed by parasite-mediated catabolism of host hemoglobin. We propose that novel PfCRT variants emerging in the field could contribute to a multigenic basis of PPQ resistance.
Rationally Designed TLR4 Ligands for Vaccine Adjuvant Discovery
Authors: Gregg Kelsey A. , Harberts Erin , Gardner Francesca M. , Pelletier Mark R. , Cayatte Corinne , Yu Li , McCarthy Michael P. , Marshall Jason D. , Ernst Robert K.
Published: May 2017

Adjuvant properties of bacterial cell wall components like MPLA (monophosphoryl lipid A) are well described and have gained FDA approval for use in vaccines such as Cervarix. MPLA is the product of chemically modified lipooligosaccharide (LOS), altered to diminish toxic proinflammatory effects while retaining adequate immunogenicity. Despite the virtually unlimited number of potential sources among bacterial strains, the number of useable compounds within this promising class of adjuvants are few. We have developed bacterial enzymatic combinatorial chemistry (BECC) as a method to generate rationally designed, functionally diverse lipid A. BECC removes endogenous or introduces exogenous lipid A-modifying enzymes to bacteria, effectively reprogramming the lipid A biosynthetic pathway. In this study, BECC is applied within an avirulent strain of
Yersinia pestis
to develop structurally distinct LOS molecules that elicit differential Toll-like receptor 4 (TLR4) activation. Using reporter cell lines that measure NF-κB activation, BECC-derived molecules were screened for the ability to induce a lower proinflammatory response than
Escherichia coli
LOS. Their structures exhibit varied, dose-dependent, TLR4-driven NF-κB activation with both human and mouse TLR4 complexes. Additional cytokine secretion screening identified molecules that induce levels of tumor necrosis factor alpha (TNF-α) and interleukin-8 (IL-8) comparable to the levels induced by phosphorylated hexa-acyl disaccharide (PHAD). The lead candidates demonstrated potent immunostimulation in mouse splenocytes, human primary blood mononuclear cells (PBMCs), and human monocyte-derived dendritic cells (DCs). This newly described system allows directed programming of lipid A synthesis and has the potential to generate a diverse array of TLR4 agonist candidates.

IMPORTANCE
There is an urgent need to develop effective vaccines against infectious diseases that continue to be major causes of morbidity and mortality worldwide. Making effective vaccines requires selecting an adjuvant to strengthen an appropriate and protective immune response. This work describes a practical method, bacterial enzymatic combinatorial chemistry (BECC), for generating functionally diverse molecules for adjuvant use. These molecules were analyzed in cell culture for their ability to initiate immune stimulatory activity. Several of the assays described herein show promising
in vitro
cytokine production and costimulatory molecule expression results, suggesting that the BECC molecules may be useful in future vaccine preparations.
Primary care services co-located with Emergency Departments across a UK region: early views on their development
Authors: Ablard Suzanne , O’Keeffe Colin , Ramlakhan Shammi , Mason Suzanne M
Published: May 2017

Background

Co-location of primary care services with Emergency Departments (ED) is one initiative aiming to reduce the burden on EDs of patients attending with non-urgent problems. However, the extent to which these services are operating within or alongside EDs is not currently known.

This study aimed to create a typology of co-located primary care services in operation across Yorkshire and Humber (Y&H) as well as identify early barriers and facilitators to their implementation and sustainability.

Methods

A self-report survey was sent to the lead consultant or other key contact at 17 EDs in the Y&H region to establish the extent and configuration of co-located primary care services. Semi-structured interviews were then conducted with urgent and unscheduled care stakeholders across five hospital sites to explore the barriers and facilitators to the formation and sustainability of these services.

Results

Thirteen EDs completed the survey and interviews were carried out with four ED consultants, one ED nurse and three general practitioners (GPs). Three distinct models were identified: ‘Primary Care Services Embedded within the ED’ (seven sites), ‘Co-located Urgent Care Centre’ (two sites) and ‘GP out-of-hours’ (nine sites). Qualitative data were analysed using framework analysis. Four interview themes emerged (justification for the service, level of integration, referral processes and sustainability) highlighting some of the challenges in implementing these co-located primary care services.

Conclusion

Creating a service within or alongside the ED in which GPs can use their distinct skills and therefore add value to the existing skill mix of ED staff is an important consideration when setting up these systems. Effective triage arrangements should also be established to ensure appropriate patients are referred to GPs. Further research is required to identify the full range of models nationally and to carry out a rigorous assessment of their impact.
Adaptation of the animal welfare assessment grid (AWAG) for monitoring animal welfare in zoological collections
Authors: Justice W. S. M. , O'Brien M. F. , Szyszka O. , Shotton J. , Gilmour J. E. M. , Riordan P. , Wolfensohn S.
Published: May 2017

Animal welfare monitoring is an essential part of zoo management and a legal requirement in many countries. Historically, a variety of welfare audits have been proposed to assist zoo managers. Unfortunately, there are a number of issues with these assessments, including lack of species information, validated tests and the overall complexity of these audits which make them difficult to implement in practice. The animal welfare assessment grid (AWAG) has previously been proposed as an animal welfare monitoring tool for animals used in research programmes. This computer‐based system was successfully adapted for use in a zoo setting with two taxonomic groups: primates and birds. This tool is simple to use and provides continuous monitoring based on cumulative lifetime assessment. It is suggested as an alternative, practical method for welfare monitoring in zoos.
Combining basin modelling with high-resolution heat-flux simulations to investigate the key drivers for burial dolomitization in an offshore carbonate reservoir
Authors: Mangione A. , Lewis H. , Geiger S. , Wood R. , Beavington-Penney S. , McQuilken J. , Cortes J.
Published: May 2017

The Eocene El Garia Formation in the offshore Hasdrubal Field was originally a nummulitic limestone in which subsequent burial dolomitization has significantly enhanced permeability. Identification of the reservoir's petrophysical property distributions requires knowledge of the spatial extent of its dolomitization, in turn requiring understanding of the processes that caused the dolomitization. Some of this understanding can be derived from measurements but others need to be simulated. In this study, the former are used as guides and we focus on the latter, evaluating the character of the dolomitizing fluid's movement and temperature patterns by using basin modelling to develop heat-flux simulations to represent the time of dolomitization. Basin modelling reconstructs the region's geology at the time of dolomitization, while heat-flux simulations recreate the appropriate conductive and convective heat and mass transport through these systems. Potential key drivers are rock mass and fault-zone permeability, and the position and shape of any salt domes. The results suggest that salt dome shape and position is the dominant control, the salt dome localizing convective systems which also use convenient faults so that hotter upwelling fluids pass through the Hasdrubal reservoir and are instrumental in the development of burial dolomitization.
Throughput Modeling of the IEEE MAC for Sensor Networks
Authors: Onderwater Martijn , Hoekstra Gerard , van der Mei Rob
Published: May 2017

In this paper we provide a model for analyzing the saturation throughput of the ieee 802.15.4 mac protocol, which is the de-facto standard for wireless sensor networks, ensuring fair access to the channel. To this end, we introduce the concept of a natural layer, which reflects the time that a sensor node typically has to wait prior to sending a packet. The model is simple and provides new insight how the throughput depends on the protocol parameters and the number of nodes in the network. Validation experiments with simulations demonstrate that the model is highly accurate for a wide range of parameter settings of the mac protocol, and applicable to both large and small networks. As a byproduct, we discuss fundamental differences in the protocol stack and corresponding throughput models of the popular 802.11 standard.
Analyzing and shaping the lifetime and the performance of barrier coverage sensor networks
Authors: Donatiello Lorenzo , Marfia Gustavo
Published: May 2017

In this work we model and provide the means to extend the lifetime of a barrier coverage sensor network deployed fortarget detection. We consider a scenario where sensors are randomly dropped on a bidimensional field in order to detect target traversals which occur in a stochastic way within a critical mission time. Once a target enters a sensor's detection area, the sensor transmits such information to a cluster head, in charge of receiving and retransmitting the messages received from the sensors deployed on the field. The contribution of this work is fourfold. We first identify the sensing nodes whose behavior is key to model the duration of sensing operations, assuming prior arrival and mobility models for target traversals. We then proceed, providing a heuristic estimation of the traffic received by the cluster head to quantify its energy requirements, resorting to specific lifetime definitions. We also evaluate the relationship between our probabilistic and heuristic models and the time until the barrier remains capable of detecting and reporting the traversal of any target to a sink, as obtained by simulation. Finally, we show how the lifetime of such network may be shaped, with the use of a sequential activation mechanism, for example to combat the traversals of adversaries exploiting the lifetime models obtained in this work.
Fusarium oxysporum and the Fusarium Wilt Syndrome
Authors: Gordon Thomas R.
Published: May 2017

The Fusarium oxysporum species complex (FOSC) comprises a multitude of strains that cause vascular wilt diseases of economically important crops throughout the world. Although sexual reproduction is unknown in the FOSC, horizontal gene transfer may contribute to the observed diversity in pathogenic strains. Development of disease in a susceptible crop requires F. oxysporum to advance through a series of transitions, beginning with spore germination and culminating with establishment of a systemic infection. In principle, each transition presents an opportunity to influence the risk of disease. This includes modifications of the microbial community in soil, which can affect the ability of pathogen propagules to survive, germinate, and infect plant roots. In addition, many host attributes, including the composition of root exudates, the structure of the root cortex, and the capacity to recognize and respond quickly to invasive growth of a pathogen, can impede development of F. oxysporum.
Network Sampling: From Snowball and Multiplicity to Respondent-Driven Sampling
Authors: Heckathorn Douglas D. , Cameron Christopher J.
Published: May 2017

Network sampling emerged as a set of methods for drawing statistically valid samples of hard-to-reach populations. The first form of network sampling, multiplicity sampling, involved asking respondents about events affecting those in their personal networks; it was subsequently applied to studies of homicide, HIV, and other topics, but its usefulness is limited to public events. Link-tracing designs employ a different approach to study hard-to-reach populations, using a set of respondents that expands in waves as each round of respondents recruit their peers. Link-tracing as applied to hidden populations, often described as snowball sampling, was initially considered a form of convenience sampling. This changed with the development of respondent-driven sampling (RDS), a widely used network sampling method in which the link-tracing design is adapted to provide the basis for statistical inference. The literature on RDS is large and rapidly expanding, involving contributions by numerous independent research groups employing data from dozens of different countries. Within this literature, many important research questions remain unresolved, including how best to choose among alternative RDS estimators, how to refine existing estimators to make them less dependent on assumptions that are sometimes counterfactual, and perhaps the greatest unresolved issue, how best to calculate the variability of the estimates.
HER2 Reactivation through Acquisition of the HER2 L755S Mutation as a Mechanism of Acquired Resistance to HER2-targeted Therapy in HER2+ Breast Cancer
Authors: Xu Xiaowei , De Angelis Carmine , Burke Kathleen A. , Nardone Agostina , Hu Huizhong , Qin Lanfang , Veeraraghavan Jamunarani , Sethunath Vidyalakshmi , Heiser Laura M. , Wang Nicholas , Ng Charlotte K.Y. , Chen Edward S. , Renwick Alexander , Wang Tao , Nanda Sarmistha , Shea Martin , Mitchell Tamika , Rajendran Mahitha , Waters Ian , Zabransky Daniel J. , Scott Kenneth L. , Gutierrez Carolina , Nagi Chandandeep , Geyer Felipe C. , Chamness Gary C. , Park Ben H. , Shaw Chad A. , Hilsenbeck Susan G. , Rimawi Mothaffar F. , Gray Joe W. , Weigelt Britta , Reis-Filho Jorge S. , Osborne C. Kent , Schiff Rachel
Published: May 2017

Purpose: Resistance to anti-HER2 therapies in HER2+ breast cancer can occur through activation of alternative survival pathways or reactivation of the HER signaling network. Here we employed BT474 parental and treatment-resistant cell line models to investigate a mechanism by which HER2+ breast cancer can reactivate the HER network under potent HER2-targeted therapies.

Experimental Design: Resistant derivatives to lapatinib (L), trastuzumab (T), or the combination (LR/TR/LTR) were developed independently from two independent estrogen receptor ER+/HER2+ BT474 cell lines (AZ/ATCC). Two derivatives resistant to the lapatinib-containing regimens (BT474/AZ-LR and BT474/ATCC-LTR lines) that showed HER2 reactivation at the time of resistance were subjected to massive parallel sequencing and compared with parental lines. Ectopic expression and mutant-specific siRNA interference were applied to analyze the mutation functionally. In vitro and in vivo experiments were performed to test alternative therapies for mutant HER2 inhibition.

Results: Genomic analyses revealed that the HER2L755S mutation was the only common somatic mutation gained in the BT474/AZ-LR and BT474/ATCC-LTR lines. Ectopic expression of HER2L755S induced acquired lapatinib resistance in the BT474/AZ, SK-BR-3, and AU565 parental cell lines. HER2L755S-specific siRNA knockdown reversed the resistance in BT474/AZ-LR and BT474/ATCC-LTR lines. The HER1/2–irreversible inhibitors afatinib and neratinib substantially inhibited both resistant cell growth and the HER2 and downstream AKT/MAPK signaling driven by HER2L755S in vitro and in vivo.

Conclusions: HER2 reactivation through acquisition of the HER2L755S mutation was identified as a mechanism of acquired resistance to lapatinib-containing HER2-targeted therapy in preclinical HER2-amplified breast cancer models, which can be overcome by irreversible HER1/2 inhibitors. Clin Cancer Res; 23(17); 5123–34. ©2017 AACR.
Head-to-Head Comparison of Inflammation and Neovascularization in Human Carotid Plaques
Authors: Demeure Fabian , Bouzin Caroline , Roelants Véronique , Bol Anne , Verhelst Robert , Astarci Parla , Gerber Bernhard L. , Pouleur Anne-Catherine , Pasquet Agnès , de Meester Christophe , Vanoverschelde Jean-Louis J. , Vancraeynest David
Published: May 2017

Background—

Inflammation and intraplaque neovascularization are acknowledged to be 2 features of plaque vulnerability, although their temporal expression and their respective value in predicting clinical events are poorly understood. To determine their respective temporal associations, we conducted a comprehensive assessment of inflammation and intraplaque neovascularization in the carotid plaque of symptomatic and asymptomatic patients.

Methods and Results—

Thirty patients with severe carotid stenosis underwent
18
F-fluorodeoxyglucose-positron emission tomography/computed tomographic imaging. Plaque
18
F-fluorodeoxyglucose-uptake, indicative of inflammation, was measured by calculating the target:background ratio. The presence of intraplaque neovascularization during contrast-enhanced ultrasound was judged semiquantitatively; low-grade contrast enhancement (CE) suggested its absence, and high-grade CE, the presence of neovascularization. Carotid surgery was performed 1.6±1.8 days after completing both imaging modalities in all patients, and the presence of macrophages and neovessels was quantified by immunohistochemistry. We identified a significant correlation between the target:background ratio and macrophage quantification (
R
=0.78;
P
<0.001). The number of vessels was also significantly higher in carotid plaque with high-CE (
P
<0.001). Surprisingly, immunohistochemistry showed that high-CE and vessel number were neither associated with an elevated target:background ratio (
P
=0.28 and
P
=0.60, respectively) nor macrophage infiltration (
P
=0.59 and
P
=0.40, respectively). Finally, macrophage infiltration and target:background ratio were higher in the carotid plaque of symptomatic patients (
P
=0.021 and
P
=0.05, respectively), whereas CE grade and the presence of neovessels were not.

Conclusions—

Inflammation and intraplaque neovascularization are not systematically associated in carotid plaques, suggesting a temporal separation between the 2 processes. Inflammation seems more pronounced when symptoms are present. These data highlight the challenges that face any imaging strategy designed to assess plaque vulnerability.
Functional Characterization of the
GUCY1A3
Coronary Artery Disease Risk Locus
Authors: Kessler Thorsten , Wobst Jana , Wolf Bernhard , Eckhold Juliane , Vilne Baiba , Hollstein Ronja , von Ameln Simon , Dang Tan An , Sager Hendrik B. , Moritz Rumpf Philipp , Aherrahrou Redouane , Kastrati Adnan , Björkegren Johan L.M. , Erdmann Jeanette , Lusis Aldons J. , Civelek Mete , Kaiser Frank J. , Schunkert Heribert
Published: May 2017

Background:

A chromosomal locus at 4q32.1 has been genome-wide significantly associated with coronary artery disease risk. The locus encompasses
GUCY1A3
, which encodes the α
1
subunit of the soluble guanylyl cyclase (sGC), a key enzyme in the nitric oxide/cGMP signaling pathway. The mechanism linking common variants in this region with coronary risk is not known.

Methods:

Gene expression and protein expression were analyzed with quantitative polymerase chain reaction and immunoblotting, respectively. Putative allele-specific transcription factors were identified with in silico analyses and validated via allele-specific quantification of antibody-precipitated chromatin fractions. Regulatory properties of the lead risk variant region were analyzed with reporter gene assays. To assess the effect of zinc finger E box-binding homeobox 1 transcription factor (ZEB1), siRNA-mediated knockdown and overexpression experiments were performed. Association of
GUCY1A3
genotype and cellular phenotypes was analyzed with vascular smooth muscle cell migration assays and platelet aggregation analyses.

Results:

Whole-blood
GUCY1A3
mRNA levels were significantly lower in individuals homozygous for the lead (rs7692387) risk variant. Likewise, reporter gene assays demonstrated significantly lower
GUCY1A3
promoter activity for constructs carrying this allele. In silico analyses located a DNase I hypersensitivity site to rs7692387 and predicted binding of the transcription factor ZEB1 rather to the nonrisk allele, which was confirmed experimentally. Knockdown of
ZEB1
resulted in more profound reduction of nonrisk allele promoter activity and a significant reduction of endogenous
GUCY1A3
expression. Ex vivo–studied platelets from homozygous nonrisk allele carriers displayed enhanced inhibition of ADP-induced platelet aggregation by the nitric oxide donor sodium nitroprusside and the phosphodiesterase 5 inhibitor sildenafil compared with homozygous risk allele carriers. Moreover, pharmacological stimulation of sGC led to reduced migration only in vascular smooth muscle cells homozygous for the nonrisk allele. In the Hybrid Mouse Diversity Panel, higher levels of
GUCY1A3
expression correlated with less atherosclerosis in the aorta.

Conclusions:

Rs7692387 is located in an intronic site that modulates
GUCY1A3
promoter activity. The transcription factor ZEB1 binds preferentially to the nonrisk allele, leading to an increase in
GUCY1A3
expression, higher sGC levels, and higher sGC activity after stimulation. Finally, human and mouse data link augmented sGC expression to lower risk of atherosclerosis.
Activation of the Amino Acid Response Pathway Blunts the Effects of Cardiac Stress
Authors: Qin Pu , Arabacilar Pelin , Bernard Roberta E. , Bao Weike , Olzinski Alan R. , Guo Yuanjun , Lal Hind , Eisennagel Stephen H. , Platchek Michael C. , Xie Wensheng , del Rosario Julius , Nayal Mohamad , Lu Quinn , Roethke Theresa , Schnackenberg Christine G. , Wright Fe , Quaile Michael P. , Halsey Wendy S. , Hughes Ashley M. , Sathe Ganesh M. , Livi George P. , Kirkpatrick Robert B. , Qu Xiaoyan A. , Rajpal Deepak K. , Faelth Savitski Maria , Bantscheff Marcus , Joberty Gerard , Bergamini Giovanna , Force Thomas L. , Gatto Gregory J. , Hu Erding , Willette Robert N.
Published: May 2017

Background

The amino acid response (
AAR
) is an evolutionarily conserved protective mechanism activated by amino acid deficiency through a key kinase, general control nonderepressible 2. In addition to mobilizing amino acids, the
AAR
broadly affects gene and protein expression in a variety of pathways and elicits antifibrotic, autophagic, and anti‐inflammatory activities. However, little is known regarding its role in cardiac stress. Our aim was to investigate the effects of halofuginone, a prolyl‐
tRNA
synthetase inhibitor, on the
AAR
pathway in cardiac fibroblasts, cardiomyocytes, and in mouse models of cardiac stress and failure.

Methods and Results

Consistent with its ability to inhibit prolyl‐
tRNA
synthetase, halofuginone elicited a general control nonderepressible 2–dependent activation of the
AAR
pathway in cardiac fibroblasts as evidenced by activation of known
AAR
target genes, broad regulation of the transcriptome and proteome, and reversal by
l
‐proline supplementation. Halofuginone was examined in 3 mouse models of cardiac stress: angiotensin
II
/phenylephrine, transverse aortic constriction, and acute ischemia reperfusion injury. It activated the
AAR
pathway in the heart, improved survival, pulmonary congestion, left ventricle remodeling/fibrosis, and left ventricular function, and rescued ischemic myocardium. In human cardiac fibroblasts, halofuginone profoundly reduced collagen deposition in a general control nonderepressible 2–dependent manner and suppressed the extracellular matrix proteome. In human induced pluripotent stem cell–derived cardiomyocytes, halofuginone blocked gene expression associated with endothelin‐1‐mediated activation of pathologic hypertrophy and restored autophagy in a general control nonderepressible 2/
eIF
2α‐dependent manner.

Conclusions

Halofuginone activated the
AAR
pathway in the heart and attenuated the structural and functional effects of cardiac stress.
Different Determinants of Ventilatory Inefficiency at Different Stages of Reduced Ejection Fraction Chronic Heart Failure Natural History
Authors: Mezzani Alessandro , Giordano Andrea , Komici Klara , Corrà Ugo
Published: May 2017

Background

It is not known whether determinants of ventilation (
VE
)/volume of exhaled carbon dioxide (
VCO
2
) slope during incremental exercise may differ at different stages of reduced ejection fraction chronic heart failure natural history.

Methods and Results

VE
/
VCO
2
slope was fitted up to lowest
VE
/
VCO
2
ratio, that is, a proxy of the
VE
/perfusion ratio devoid of nonmetabolic stimuli to ventilatory drive.
VE
/
VCO
2
slope tertiles were generated from our database (<27.5 [tertile 1], ≥27.5 to <32.0 [tertile 2], and ≥32.0 [tertile 3]), and 147 chronic heart failure patients with repeated tests yielding
VE
/
VCO
2
slopes in 2 different tertiles were selected. Determinants of
VE
/
VCO
2
slope changes across tertile pairs 1 versus 2, 2 versus 3, and 1 versus 3 were assessed by exploring changes in
VE
and
VCO
2
at lowest
VE
/
VCO
2
and those in
VE
/work rate (W) and
VCO
2
/W slope. Resting and peak cardiac output (
CO
) were calculated as
VO
2
/estimated arteriovenous O
2
difference and the
CO
/W slope analyzed. Notwithstanding a progressively lower W with increasing tertile,
VE
at lowest
VE
/
VCO
2
and
VE
/W slope were significantly higher in tertiles 2 and 3 versus tertile 1. Conversely,
VCO
2
at lowest
VE
/
VCO
2
and
CO
/W slope significantly decreased across tertiles, whereas
VCO
2
/W slope did not. Difference (Δ) in
VE
/W slope between tertiles accounted for 71% of Δ
VE
/
VCO
2
slope variance, with Δ
VCO
2
/W slope explaining an additional 26% (model
r
=0.99;
r
2
=0.97;
P
<0.0001). Similar results were obtained substituting Δ
VCO
2
/W slope with Δ
CO
/W slope.

Conclusions

Ventilatory overactivation is the predominant cause of
VE
/
VCO
2
slope increase at initial stages of chronic heart failure, whereas hemodynamic impairment plays an additional role at more‐advanced pathophysiological stages.
Enhanced Detection of Edema in Malignant Anterior Circulation Stroke (EDEMA) Score
Authors: Ong Charlene Jennifer , Gluckstein Jeffrey , Laurido-Soto Osvaldo , Yan Yan , Dhar Rajat , Lee Jin-Moo
Published: May 2017

Background and Purpose—

Rapid recognition of those at high risk for malignant edema after stroke would facilitate triage for monitoring and potential surgery. Admission data may be insufficient for accurate triage decisions. We developed a risk prediction score using clinical and radiographic variables within 24 hours of ictus to better predict potentially lethal malignant edema.

Methods—

Patients admitted with diagnosis codes of cerebral edema and ischemic stroke, NIHSS score (National Institute of Health Stroke Score) of ≥8 and head computed tomographies within 24 hours of stroke onset were included. Primary outcome of potentially lethal malignant edema was defined as death with midline shift ≥5 mm or decompressive hemicraniectomy. We performed multivariate analyses on data available within 24 hours of ictus. Bootstrapping was used to internally validate the model, and a risk score was constructed from the results.

Results—

Thirty-three percent of 222 patients developed potentially lethal malignant edema. The final model C statistic was 0.76 (confidence interval, 0.68–0.82) in the derivation cohort and 0.75 (confidence interval, 0.72–0.77) in the bootstrapping validation sample. The EDEMA score (Enhanced Detection of Edema in Malignant Anterior Circulation Stroke) was developed using the following independent predictors: basal cistern effacement (=3); glucose ≥150 (=2); no tPA (tissue-type plasminogen activator) or thrombectomy (=1), midline shift >0 to 3 (=1), 3 to 6 (=2), and 6 to 9 (=4); >9 (=7); and no previous stroke (=1). A score over 7 was associated with 93% positive predictive value.

Conclusions—

The EDEMA score identifies patients at high risk for potentially lethal malignant edema. Although it requires external validation, this scale could help expedite triage decisions in this patient population.
ASPECTS (Alberta Stroke Program Early CT Score) Measurement Using Hounsfield Unit Values When Selecting Patients for Stroke Thrombectomy
Authors: Mokin Maxim , Primiani Christopher T. , Siddiqui Adnan H. , Turk Aquilla S.
Published: May 2017

Background and Purpose—

The ASPECTS (Alberta Stroke Program Early CT Score) is a quantitate score that measures the extent of early ischemic changes. Our aim was to investigate how measurement of ASPECTS using Hounsfield unit (HU) values on initial noncontrast head computerized tomography (CT) correlates with the extent of final infarct on follow-up imaging.

Methods—

Cases of acute stroke from the middle cerebral artery M1 occlusion in which complete recanalization (TICI [Thrombolysis in Cerebral Infarction] 3) was achieved were included for analysis. Using HU ratio (HU affected/HU control hemisphere) and HU difference (HU control−HU affected hemisphere) values, ASPECTS was measured on initial CT imaging and correlated with final ASPECTS at 24 hours. The study cohort consisted of 41 patients with acute stroke from the M1 occlusion. The mean time from stroke symptoms onset to baseline head CT imaging was 264 minutes and from CT to TICI 3 recanalization was 142 minutes.

Results—

HU ratio within the 0.94 to 0.96 ranges showed the highest correlation coefficient and lowest mean and median errors with the final ASPECTS. The difference of 2.0 HU between the 2 hemispheres demonstrated the higher correlation coefficient (
r
=0.71;
P
<0.0001) and the lowest mean and median absolute errors (1.4 and 1, respectively).

Conclusions—

We established a simple algorithm for rapid and accurate assessment of ASPECTS on baseline CT imaging to predict the extent of final stroke in patients with emergent large vessel occlusion who undergo endovascular revascularization.
Hemorrhage Risk of Brain Arteriovenous Malformations During Pregnancy and Puerperium in a North American Cohort
Authors: Porras Jose L. , Yang Wuyang , Philadelphia Eunice , Law Jody , Garzon-Muvdi Tomas , Caplan Justin M. , Colby Geoffrey P. , Coon Alexander L. , Tamargo Rafael J. , Huang Judy
Published: May 2017

Background and Purpose—

Conclusions reached in existing literature about risk of arteriovenous malformation (AVM) hemorrhage during pregnancy are controversial. This study compares the risk of hemorrhage in pregnant and nonpregnant female patients with AVM in a North American cohort.

Methods—

We retrospectively reviewed female patients with AVM evaluated from 1990 to 2015. Exposure period for pregnancy and puerperium was calculated as 40 and 6 weeks, respectively, for each full-term pregnancy and 6 weeks for each abortion. Hemorrhage events and patient-years were calculated during an exposure period (pregnancy and puerperium), and a nonexposure period defined as either the interval from birth until AVM obliteration or until last follow-up after subtracting exposure period. Poisson rate ratio test was used to compare rate of hemorrhage between exposure and nonexposure periods.

Results—

For 270 female patients with AVM, mean age was 35.0±19.6 years, and race distribution was white (n=165, 61.1%), black (n=59, 21.9%), Hispanic (n=9, 3.3%), Asian (n=6, 2.2%), and other (n=31, 11.5%). From 191 total pregnancies occurring before AVM obliteration, there were 175 (91.6%) term deliveries and 16 (8.4%) abortions. Overall annual hemorrhage rate for 149 total hemorrhages during an average of 11 097 patient-years was 1.34%. There were 140 hemorrhages in nonexposed women and 9 hemorrhages in pregnant women, translating to an annual hemorrhage rate of 1.3% in nonpregnant women versus 5.7% in pregnant women (
P
<0.001). Identical analysis for reproductive age patients (15–50) demonstrated a rate of 1.3% versus 7.0% (
P
<0.001).

Conclusions—

Our results conflict with those from a recent study describing no increased rate of rupture during pregnancy. This difference may reflect unique population attributes influencing brain AVM hemorrhage during pregnancy.
Reading the Bible as a Feminist
Authors: Koosed Jennifer L.
Published: May 2017

This work provides a brief introduction to feminist interpretation of scripture. Feminist interpretation is first grounded in feminism as an intellectual and political movement. Next, this introduction briefly recounts the origins of feminist readings of the Bible with attention to both early readings and the beginnings of feminist biblical scholarship in the academy. Feminist biblical scholarship is not a single methodology, but rather an approach that can shape any reading method. As a discipline, it began with literary-critical readings (especially of the Hebrew Bible) but soon also broached questions of women’s history (especially in the New Testament and Christian origins). Since these first forays, feminist interpretation has influenced almost every type of biblical scholarship. The third section of this essay, then, looks at gender archaeology, feminist poststructuralism and postcolonial readings, and newer approaches informed by gender and queer theory. Finally, it ends by examining feminist readings of Eve.