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Purpose: To characterize the retinal phenotype of juvenile neuronal ceroid lipofuscinosis (JNCL), highlight delayed and mistaken diagnosis, and propose an algorithm for early identification., Design: Retrospective case series., Participants: Eight children (5 female) with JNCL., Methods: Review of clinical notes, retinal imaging including fundus autofluorescence and OCT, electroretinography (ERG), and both microscopy and molecular genetic testing., Main Outcome Measurements: Demographic data, signs and symptoms, visual acuity (VA), fundus autofluorescence and OCT findings, ERG phenotype, and microscopy/molecular genetics., Results: Participants presented with rapid bilateral vision loss over 1 to 18 months, with mean VA deteriorating from 0.44 logarithm of the minimum angle of resolution (logMAR) (range, 0.20-1.78 logMAR) at baseline to 1.34 logMAR (0.30 logMAR - light perception) at last follow-up. Age of onset ranged from 3 to 7 years (mean, 5.3 years). The age at diagnosis of JNCL ranged from 7 to 10 years (mean, 8.3 years). Six children displayed eccentric fixation, and 6 children had cognitive or neurologic signs at the time of diagnosis (75%). Seven patients had bilateral bull's-eye maculopathy at presentation. Coats-like exudative vasculopathy, not previously reported in JNCL, was observed in 1 patient. OCT imaging revealed near complete loss of outer retinal layers and marked atrophy of the nerve fiber and ganglion cell layers at the central macula. An electronegative ERG was present in 4 patients (50%), but with additional a-wave reduction, there was an undetectable ERG in the remaining 4 patients. Blood film microscopy revealed vacuolated lymphocytes, and electron microscopy showed lysosomal (fingerprint) inclusions in all 8 patients., Conclusions: In a young child with bilateral rapidly progressive vision loss and macular disturbance, blood film microscopy to detect vacuolated lymphocytes is a rapid, readily accessible, and sensitive screening test for JNCL. Early suspicion of JNCL can be aided by detailed directed history and high-resolution retinal imaging, with subsequent targeted microscopy/genetic testing. Early diagnosis is critical to ensure appropriate management, counseling, support, and social care for children and their families. Furthermore, although potential therapies for this group of disorders are in early-phase clinical trial, realistic expectations are that successful intervention will be most effective when initiated at the earliest stage of disease., (Copyright © 2019 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.)

Intramyocardial hemorrhage is an independent predictor of adverse outcomes in ST-segment elevation myocardial infarction (STEMI). Iron deposition resulting from ischemia-reperfusion injury (I/R) is pro-inflammatory and has been associated with adverse remodeling. The role of iron chelation in hemorrhagic acute myocardial infarction (AMI) has never been explored. The purpose of this study was to investigate the cardioprotection offered by the iron-chelating agent deferiprone (DFP) in a porcine AMI model by evaluating hemorrhage neutralization and subsequent cardiac remodeling. Two groups of animals underwent a reperfused AMI procedure: control and DFP treated (N = 7 each). A comprehensive MRI examination was performed in healthy state and up to week 4 post-AMI, followed by histological assessment. Infarct size was not significantly different between the two groups; however, the DFP group demonstrated earlier resolution of hemorrhage (by T2* imaging) and edema (by T2 imaging). Additionally, ventricular enlargement and myocardial hypertrophy (wall thickness and mass) were significantly smaller with DFP, suggesting reduced adverse remodeling, compared to control. The histologic results were consistent with the MRI findings. To date, there is no effective targeted therapy for reperfusion hemorrhage. Our proof-of-concept study is the first to identify hemorrhage-derived iron as a therapeutic target in I/R and exploit the cardioprotective properties of an iron-chelating drug candidate in the setting of AMI. Iron chelation could potentially serve as an adjunctive therapy in hemorrhagic AMI.

PD-L1 is expressed in tumor cells and its interaction with PD-1 plays an important role in evading immune surveillance; this can be overcome using PD-L1 or PD-1 immunotherapy antibodies. This study reports a novel approach for targeting PD-L1. In human breast cancer cell lines and 4T1 mouse mammary tumor cells, PD-L1 expression was regulated by the nuclear receptor NR4A1/Sp1 complex bound to the proximal germinal center (GC)-rich region of the PD-L1 gene promoter. Treatment of breast cancer cells with bis-indole-derived NR4A1 antagonists including 1,1-bis(3'-indolyl)-1-(3-chloro-4-hydroxy-5-methoxyphenyl)methane (Cl-OCH3) decreased expression of PD-L1 mRNA, promoter-dependent luciferase activity, and protein. In in vivo studies using a syngeneic mouse model bearing orthotopically injected 4T1 cells, Cl-OCH3 decreased tumor growth and weight and inhibited lung metastasis. Cl-OCH3 also decreased expression of CD3 + /CD4 + /CD25 + /FoxP3 + regulatory T cells and increased the Teff/Treg ratio. Therefore, the potent anticancer activities of NR4A1 antagonists are also accompanied by enhanced antitumor immunity in PD-L1-expressing triple-negative breast cancer and thus represent a novel class of drugs that mimic immunotherapy. SIGNIFICANCE: These findings show that the orphan nuclear receptor NR4A1 controls PD-L1 expression and identify a chemical probe capable of disrupting this regulatory axis., (©2020 American Association for Cancer Research.)

Neonicotinoids are pesticides used to protect crops but with known secondary influences at sublethal doses on bees. Honeybees use their sense of smell to identify the queen and nestmates, to signal danger and to distinguish flowers during foraging. Few behavioural studies to date have examined how neonicotinoid pesticides affect the ability of bees to distinguish odours. Here, we used a differential learning task to test how neonicotinoid exposure affects learning, memory and olfactory perception in foraging-age honeybees. Bees fed with thiamethoxam could not perform differential learning and could not distinguish odours during short- and long-term memory tests. Our data indicate that thiamethoxam directly impacts the cognitive processes involved in working memory required during differential olfactory learning. Using a combination of behavioural assays, we also identified that thiamethoxam has a direct impact on the olfactory perception of similar odours. Honeybees fed with other neonicotinoids (clothianidin, imidacloprid, dinotefuran) performed the differential learning task, but at a slower rate than the control. These bees could also distinguish the odours. Our data are the first to show that neonicotinoids have compound specific effects on the ability of bees to perform a complex olfactory learning task. Deficits in decision making caused by thiamethoxam exposure could mean that this is more harmful than other neonicotinoids, leading to inefficient foraging and a reduced ability to identify nestmates., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2020. Published by The Company of Biologists Ltd.)

GABA is the most common inhibitory neurotransmitter in both vertebrate and invertebrate nervous systems. In insects, inhibition plays important roles at the neuromuscular junction, in the regulation of central pattern generators, and in the modulation of information in higher brain processing centers. Additionally, increasing our understanding of the functions of GABA is important since GABA A receptors are the targets of several classes of pesticides. To investigate the role of GABA in motor function, honey bee foragers were injected with GABA or with agonists or antagonists specific for either GABA A or GABA B receptors. Compounds that activated either type of GABA receptor decreased activity levels. Bees injected with the GABA A receptor antagonist picrotoxin lost the ability to right themselves, whereas blockade of GABA B receptors led to increases in grooming. Injection with antagonists of either GABA A or GABA B receptors resulted in an increase in extended wing behavior, during which bees kept their wings out at right angles to their body rather than folded along their back. These data suggest that the GABA receptor types play distinct roles in behavior and that GABA may affect behavior at several different levels., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Snakebite envenoming is a serious and neglected tropical disease that kills ~100,000 people annually. High-quality, genome-enabled comprehensive characterization of toxin genes will facilitate development of effective humanized recombinant antivenom. We report a de novo near-chromosomal genome assembly of Naja naja, the Indian cobra, a highly venomous, medically important snake. Our assembly has a scaffold N50 of 223.35 Mb, with 19 scaffolds containing 95% of the genome. Of the 23,248 predicted protein-coding genes, 12,346 venom-gland-expressed genes constitute the 'venom-ome' and this included 139 genes from 33 toxin families. Among the 139 toxin genes were 19 'venom-ome-specific toxins' (VSTs) that showed venom-gland-specific expression, and these probably encode the minimal core venom effector proteins. Synthetic venom reconstituted through recombinant VST expression will aid in the rapid development of safe and effective synthetic antivenom. Additionally, our genome could serve as a reference for snake genomes, support evolutionary studies and enable venom-driven drug discovery.

Purpose: Familial exudative vitreoretinopathy (FEVR) is a rare finding in patients with genetic forms of microcephaly. This study documents the detailed phenotype and expands the range of genetic heterogeneity., Design: Retrospective case series., Methods: Twelve patients (10 families) with a diagnosis of FEVR and microcephaly were ascertained from pediatric genetic eye clinics and underwent full clinical assessment including retinal imaging. Molecular investigations included candidate gene Sanger sequencing, whole-exome sequencing (WES), and whole-genome sequencing (WGS)., Results: All patients had reduced vision and nystagmus. Six were legally blind. Two probands carried bi-allelic LRP5 variants, both presenting with bilateral retinal folds. A novel homozygous splice variant, and 2 missense variants were identified. Subsequent bone density measurement identified osteoporosis in one proband. Four families had heterozygous KIF11 variants. Two probands had a retinal fold in one eye and chorioretinal atrophy in the other; the other 2 had bilateral retinal folds. Four heterozygous variants were found, including 2 large deletions not identified on Sanger sequencing or WES. Finally, a family of 2 children with learning difficulties, abnormal peripheral retinal vasculogenesis, and rod-cone dystrophy were investigated. They were found to have bi-allelic splicing variants in TUBGCP6. Three families remain unsolved following WES and WGS., Conclusions: Molecular diagnosis has been achieved in 7 of 10 families investigated, including a previously unrecognized association with LRP5. WGS enabled molecular diagnosis in 3 families after prior negative Sanger sequencing of the causative gene. This has enabled patient-specific care with targeted investigations and accurate family counseling., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Background: The promise of biopharmaceuticals comprising one or more binding domains motivates the development of novel methods for de novo isolation and affinity maturation of virion-binding domains. Identifying avenues for overcoming the challenges associated with using virions as screening reagents is paramount given the difficulties associated with obtaining high-purity virus-associated proteins that retain the conformation exhibited on the virion surface., Results: Fluorescence activated cell sorting (FACS) of 1.5 × 10 7 clones taken from a naïve yeast surface-displayed human fibronectin domain (Fn3) against whole virions yielded two unique binders to Zika virions. Construction and FACS of site-directed binding loop mutant libraries based on one of these binders yielded multiple progeny clones with enhanced Zika-binding affinities. These affinity-matured clones bound Zika virions with low double- or single-digit nanomolar affinity in ELISA assays, and expressed well as soluble proteins in E. coli shake flask culture, with post-purification yields exceeding 10 mg/L., Conclusions: FACS of a yeast-displayed binding domain library is an efficient method for de novo isolation of virion-binding domains. Affinities of isolated virion-binding clones are readily enhanced via FACS screening of mutant progeny libraries. Given that most binding domains are compatible with yeast display, the approach taken in this work may be broadly utilized for generating virion-binding domains against many different viruses for use in passive immunotherapy and the prevention of viral infection., Competing Interests: Competing interestsThe authors declare that they have no competing interests., (© The Author(s). 2019.)

Introduction: Cardiac magnetic resonance imaging (CMR) is the gold standard for the assessment of left ventricular (LV) function. However, traditional sequences are time-consuming and require breath-holding. Our aim was to evaluate the image quality of LV functional assessment with a novel, accelerated, free-breathing cine sequence and to compare LV functional parameters between it and a traditional sequence., Methods: This was a prospective cohort study of 31 patients with cardiomyopathy. All studies were performed on a 1.5 Tesla scanner. LV function was first determined using contiguous short axis slices covering the left ventricle from the base to the apex acquired with the standard cine sequence. Next, the accelerated sequence was acquired for each patient. The Wilcoxon Matched-Pair Sign Rank Test was used to compare image quality between the accelerated and traditional cine imaging sequences. Standard and accelerated left ventricular volumes and ejection fraction were compared using linear regression. Bland-Altman plots were then constructed to evaluate agreement, interobserver and intraobserver variability for left ventricular volumes and ejection fraction., Results: Mean acquisition time was 29 s for the accelerated sequence vs. 410 s for the traditional sequence. Qualitative assessment of image quality was similar for both sequences (p = 0.23). There were no significant differences in terms of LVEDV, LVESV, LVSV, LV mass and LVEF when calculated from either sequence with very good agreement between the standard and accelerated sequences. The mean differences with 95% limits of agreement were as follows: LV mass (-0.6, -22.9 g, 21.6 g), LVEDV (5.1 mL, -18.4 mL, 28.9 mL), LVEF (-0.3, -5.4, 4.7), LVESV (4.0 mL, -12.0 mL, 20.0 mL), LVSV (1.1 mL, -13.3 mL, 15.5 mL). Interobserver variability ranged from 0.1 to 6.3% while intraobserver variability ranged from 0.1 to 1.8%., Conclusions: The accelerated free-breathing cine sequence performed similarly to standard of care multi breath-hold cine imaging and was acquired in a fraction of the time without the need for breath-holding. If applied to clinical practice, this sequence can significantly reduce scanning time and facilitate CMR scanning in those patients who are unable to breath-hold., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Multicomponent therapy has gained interest for its potential to synergize and subsequently lower the effective dose of each constituent required to reduce colon cancer risk. We have previously showed that rapidly cycling Lgr5 stem cells are exquisitely sensitive to extrinsic dietary factors that modulate colon cancer risk. In the present study, we quantified the dose-dependent synergistic properties of dietary n-3 polyunsaturated fatty acids (PUFA) and curcumin (Cur) to promote targeted apoptotic deletion of damaged colonic Lgr5 stem cells. For this purpose, both heterogeneous bulk colonocytes and Lgr5 stem cells were isolated from Lgr5-EGFP-IRES-CreER knock-in mice injected with azoxymethane (AOM). Isolated cells were analyzed for DNA damage (γH2AX), apoptosis (cleaved caspase-3), and targeted apoptosis (both γH2AX and cleaved caspase-3) at 12 h post-AOM injection. Comparison of the percentage of targeted apoptosis in Lgr5 stem cells (GFP) across a broad bioactive dose-range revealed an ED50 of 16.0 mg/day n-3 PUFA + 15.9 mg/day Cur. This corresponded to a human equivalent dose of 3.0 g n-3 PUFA + 3.0 g Cur. In summary, our results provide evidence that a low dose (n-3 PUFA + Cur) combination diet reduces AOM-induced DNA damage in Lgr5 stem cells and enhances targeted apoptosis of DNA-damaged cells, implying that a lower human equivalent dose can be utilized in future human clinical trials.

The development of high-quality chromosomally assigned reference genomes constitutes a key feature for understanding genome architecture of a species and is critical for the discovery of the genetic blueprints of traits of biological significance. South American camelids serve people in extreme environments and are important fiber and companion animals worldwide. Despite this, the alpaca reference genome lags far behind those available for other domestic species. Here we produced a chromosome-level improved reference assembly for the alpaca genome using the DNA of the same female Huacaya alpaca as in previous assemblies. We generated 190X Illumina short-read, 8X Pacific Biosciences long-read and 60X Dovetail Chicago ® chromatin interaction scaffolding data for the assembly, used testis and skin RNAseq data for annotation, and cytogenetic map data for chromosomal assignments. The new assembly VicPac3.1 contains 90% of the alpaca genome in just 103 scaffolds and 76% of all scaffolds are mapped to the 36 pairs of the alpaca autosomes and the X chromosome. Preliminary annotation of the assembly predicted 22,462 coding genes and 29,337 isoforms. Comparative analysis of selected regions of the alpaca genome, such as the major histocompatibility complex (MHC), the region involved in the Minute Chromosome Syndrome (MCS) and candidate genes for high-altitude adaptations, reveal unique features of the alpaca genome. The alpaca reference genome VicPac3.1 presents a significant improvement in completeness, contiguity and accuracy over VicPac2 and is an important tool for the advancement of genomics research in all New World camelids.

Human embryonic stem cell-derived cardiomyocytes (hESC-CMs) show considerable promise for regenerating injured hearts, and we therefore tested their capacity to stably engraft in a translationally relevant preclinical model, the infarcted pig heart. Transplantation of immature hESC-CMs resulted in substantial myocardial implants within the infarct scar that matured over time, formed vascular networks with the host, and evoked minimal cellular rejection. While arrhythmias were rare in infarcted pigs receiving vehicle alone, hESC-CM recipients experienced frequent monomorphic ventricular tachycardia before reverting back to normal sinus rhythm by 4 weeks post transplantation. Electroanatomical mapping and pacing studies implicated focal mechanisms, rather than macro-reentry, for these graft-related tachyarrhythmias as evidenced by an abnormal centrifugal pattern with earliest electrical activation in histologically confirmed graft tissue. These findings demonstrate the suitability of the pig model for the preclinical development of a hESC-based cardiac therapy and provide new insights into the mechanistic basis of electrical instability following hESC-CM transplantation., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

Aim: Intensified haemodialysis is associated with regression of left ventricular (LV) mass. Compared to LV ejection fraction, LV strain allows more direct assessment of LV function. We sought to assess the impact of in-centre nocturnal haemodialysis (INHD) on global LV strain (radial, circumferential, and longitudinal) and torsion by cardiac MRI (CMR)., Methods: In this prospective, two-centre cohort study, 37 participants on conventional haemodialysis (CHD, 3-4 h/session for three sessions/week) converted to INHD (7-8 h/session for three sessions/week) and 30 participants continued CHD. Participants underwent CMR using a standardized protocol and had biomarker measurements at baseline and 52 weeks., Results: Among the 55 participants (mean age 55; 40% women) with complete CMR data, those who converted to INHD had a significant improvement in their global circumferential strain (GCS, P = 0.025), while those continuing CHD did not have any significant changes in LV strain. When the two groups were compared, there was significant improvement in torsion. LV strains were significantly correlated with each other, but not with troponin I, C-reactive protein, or brain natriuretic protein (NT-proBNP), except for global longitudinal strain (GLS) with troponin I (P = 0.001) and NT-proBNP (P = 0.038)., Conclusion: Conversion to INHD was associated with significant improvement in GCS over one year of study, although comparisons with the CHD group were not significant. There was also a significant decrease in torsion in the INHD group compared with CHD. Improvement in LV regional function would support the notion that INHD has favourable effects on both LV structure and function., (© 2018 Asian Pacific Society of Nephrology.)

Myocardial infarct (MI) related indices determined by late gadolinium enhancement (LGE) MRI have been widely investigated in determining patients suitable for implantable cardiovascular-defibrillator (ICD) therapy to complement left ventricular ejection fraction (LV EF). In comparison to LGE-MRI using inversion-recovery fast-gradient-echo (IR-FGRE), T1 mapping techniques, such as multi contrast late enhancement (MCLE), have been shown to provide more quantitative and reproducible estimates of infarct regions. The objective of this study is to use individualized heart computer models in determining the efficacy of IR-FGRE and MCLE techniques in predicting the occurrence of post-MI ventricular tachycardia (VT). Twenty-seven patients with MI underwent LGE-MRI using IR-FGRE and MCLE prior to ICD implantation and were followed up for 6-46 months. Individualized image-based computational models were built separately for each imaging technique; simulations of propensity to VT were conducted with each model. The imaging methods were evaluated by comparing simulated inducibility of VT to clinical outcome (appropriate ICD therapy) in patients. Twelve patients had at least one appropriate ICD therapy for VT at follow-up. For both MCLE and IR-FGRE, the outcomes of the simulations of VT were significantly associated with the events of appropriate ICD therapy. This indicates that, as compared to conventional measurements such as LV EF, the simulations of VT corresponding to both MCLE and IR-FGRE were more sensitive in predicting appropriate ICD therapy in post-MI patients.

Purpose: Most MR-guided catheter-based procedures, and imaging of patients with implanted medical devices, are currently contraindicated due to a significant risk of heating associated with induced RF currents. The induced RF current produces a corresponding artifact which can be used to remotely characterize current and safely predict RF heating. Application of this remote technique in vivo to safely quantify RF heating risk may allow for execution of many scans currently contraindicated. Sources of phase other than induced RF current may present difficulty in practical in vivo., Methods: A custom ultra-short echo time (UTE) sequence was developed to minimize unwanted phase contributions. A phantom experiment was performed to compare current characterization using a stock gradient-echo (GRE) sequence and the custom UTE sequence following calibration of the temperature measurement apparatus using a previously published heating prediction technique. Animal experiments were used to investigate the feasibility of using the UTE sequence to quantify RF heating., Results: Current characterization and heating prediction with a stock GRE sequence was equivalent to that with the custom UTE sequence. Heating measurements and image-based predictions in animal experiments agreed within error in all experiments., Conclusion: Through comparison of measured heating and image-based prediction, feasibility of using a custom UTE sequence to quantify RF heating risk in vivo was demonstrated., (© 2018 International Society for Magnetic Resonance in Medicine.)

Objectives: Cardiovascular magnetic resonance (CMR) imaging is the gold-standard test for the assessment of heart function. Despite its importance, many jurisdictions lack specific billing codes that can be used to identify patient receipt of CMR in administrative databases, limiting the ability to perform 'big data' CMR studies. Our objective was to identify the optimal billing code combination to identify patients who underwent CMR using administrative data in Ontario., Design: Retrospective cohort study., Setting: Quaternary care academic referral centre in Ontario, Canada., Participants: We tested all billing code combinations in order to identify the optimal one to determine receipt of CMR. The reference gold standard was a list of all cardiothoracic magnetic resonance scans performed at Sunnybrook Health Sciences Centre between 1 January 2014 and 31 December 2016, verified by chart audit. We assessed the diagnostic performance (accuracy, sensitivity, specificity, positive predictive value and negative predictive value) for all code combinations., Results: Our gold-standard cohort consisted of 2339 thoracic MRIs that were performed at Sunnybrook Health Sciences Centre from 1 January 2014 to 31 December 2016. Of these, 2139 (91.5%) were CMRs and 200 (8.5%) were chest MRIs. We identified the most accurate billing combination for the determination of patient receipt of CMR. This combination resulted in an accuracy of 95.3% (95% CI 94.4% to 96.2%), sensitivity of 97.4% (95% CI 96.6% to 98.1%), specificity of 86.4% (95% CI 83.1% to 89.6%), positive predictive value of 96.9% (95% CI 96.1% to 97.6%) and negative predictive value of 88.4% (95% CI 85.4% to 91.5%)., Conclusions: Our study is the first to verify the ability to accurately identify patient receipt of CMR using administrative data, facilitating more robust population-based CMR studies in the future., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2018. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Despite recent improvements in the engineering of viral envelope proteins, it remains a significant challenge to create lentiviral vectors that allow targeted transduction to specific cell populations of interest. In this study, we developed a simple 'plug and play' strategy to retarget lentiviral vectors to any desired cell types through in vitro covalent modification of the virions with specific cell-targeting proteins (CTPs). This strategy exploits a disulfide bond-forming protein-peptide pair PDZ1 and its pentapeptide ligand (ThrGluPheCysAla, TEFCA). PDZ1 was incorporated into an engineered Sindbis virus envelope protein (Sind-PDZ1) and displayed on lentiviral particles while the TEFCA pentapeptide ligand was genetically linked to the CTP. Her2/neu-binding designed ankyrin repeat proteins (DARPin) were used as our model CTPs. DARPin-functionalized unconcentrated lentiviral vectors harboring Sind-PDZ1 envelope protein (Sind-PDZ1-pp) exhibited >800-fold higher infectious titer in HER2 + cells than the unfunctionalized virions (8.5 × 10 6 vs. <10 4 IU/mL). Moreover, by virtue of the covalent disulfide bond interaction between PDZ1 and TEFCA, the association of the CTP with the virions is nonreversible under non-reducing conditions (e.g. serum), making these functionalized virions potentially stable in an in vivo setting.

Background: Following acute myocardial infarction (AMI), microvascular integrity and function may be compromised as a result of microvascular obstruction (MVO) and vasodilator dysfunction. It has been observed that both infarcted and remote myocardial territories may exhibit impaired myocardial blood flow (MBF) patterns associated with an abnormal vasodilator response. Arterial spin labeled (ASL) CMR is a novel non-contrast technique that can quantitatively measure MBF. This study investigates the feasibility of ASL-CMR to assess MVO and vasodilator response in swine., Methods: Thirty-one swine were included in this study. Resting ASL-CMR was performed on 24 healthy swine (baseline group). A subset of 13 swine from the baseline group underwent stress ASL-CMR to assess vasodilator response. Fifteen swine were subjected to a 90-min left anterior descending (LAD) coronary artery occlusion followed by reperfusion. Resting ASL-CMR was performed post-AMI at 1-2 days (N = 9, of which 6 were from the baseline group), 1-2 weeks (N = 8, of which 4 were from the day 1-2 group), and 4 weeks (N = 4, of which 2 were from the week 1-2 group). Resting first-pass CMR and late gadolinium enhancement (LGE) were performed post-AMI for reference., Results: At rest, regional MBF and physiological noise measured from ASL-CMR were 1.08 ± 0.62 and 0.15 ± 0.10 ml/g/min, respectively. Regional MBF increased to 1.47 ± 0.62 ml/g/min with dipyridamole vasodilation (P < 0.001). Significant reduction in MBF was found in the infarcted region 1-2 days, 1-2 weeks, and 4 weeks post-AMI compared to baseline (P < 0.03). This was consistent with perfusion deficit seen on first-pass CMR and with MVO seen on LGE. There were no significant differences between measured MBF in the remote regions pre and post-AMI (P > 0.60)., Conclusions: ASL-CMR can assess vasodilator response in healthy swine and detect significant reduction in regional MBF at rest following AMI. ASL-CMR is an alternative to gadolinium-based techniques for assessment of MVO and microvascular integrity within infarcted, as well as salvageable and remote myocardium. This has the potential to provide early indications of adverse remodeling processes post-ischemia.

Endovascular wires and devices for peripheral arterial disease therapy have evolved greatly, yet failure rates of these procedures remain high. Information on lesion composition may inform device selection to improve the success rates of these procedures. This paper, presents an approach for informed guidewire selection. The objective of this study is to quantitatively assess the performance of a radiofrequency powered guidewire in the crossing of various morphology types of peripheral chronic total occlusions. Samples taken from amputated patient limbs are characterized by magnetic resonance imaging. Using a customized catheter test station, the performance of a radiofrequency powered guidewire in puncturing these lesions is compared to a conventional guidewire, and to itself when not powered. The analysis includes quantitative and statistical comparisons of the puncture forces experienced by the different guidewires in "hard" vs. "soft" lesions as well as qualitative assessment of deflections, buckling and puncture success of the wires. Results indicate that the use of radiofrequency ablation significantly reduces the required puncture force, reduced events of buckling and deflection, and resulted in a significantly higher puncture success rate.

Objective: Limitations with current peripheral arterial imaging modalities make selection of patients for percutaneous vascular interventions difficult. The purpose of this study was to determine whether a novel preprocedural magnetic resonance imaging (MRI) method can identify lesions that would be more challenging to cross during percutaneous vascular intervention., Methods: Fourteen patients with peripheral arterial disease underwent MRI before their intervention. A novel steady-state free precession flow-independent magnetic resonance (MR) angiogram was used to locate lesions, and an ultrashort echo time image was used to characterize hard lesion components including calcium and dense collagen. Lesions were characterized as hard if ≥50% of the lumen was occluded with calcium or collagen (as determined by MR image characteristics) in the hardest cross section within the lesion. The primary outcome was the time it took to cross a guidewire through the target lesion. The secondary outcome was the need for stenting., Results: Of 14 lesions, 8 (57%) were defined as hard and 6 (43%) were soft on the basis of MR image characteristics. Hard lesions took significantly longer to cross than soft lesions (average, 14 minutes 49 seconds vs 2 minutes 17 seconds; P = .003). Hard lesions also required stenting more often than soft lesions (Fisher exact test, P = .008). Of 14 lesions, 2 (14%) could not be crossed with a guidewire, and both lesions were hard. MR images also detected occult patencies and noncalcified hard lesions that could not be seen on X-ray angiography., Conclusions: MRI can be used to determine which peripheral arterial lesions are more difficult to cross with a guidewire. Future work will determine whether MRI lesion characterization can predict long-term endovascular outcomes to aid in procedure planning., (Copyright © 2017 Society for Vascular Surgery. Published by Elsevier Inc. All rights reserved.)

Purpose: To achieve consistent effectiveness in reconstruction of fine image features for cases of varying contrast-to-noise ratio (CNR) to facilitate translating accelerated multicontrast volumetric imaging with isotropic resolution toward clinical utility in peri-infarct characterization., Theory and Methods: A low-rank and spatially varying edge-preserving constrained compressed sensing parallel imaging reconstruction method (CP-LASER) is developed to effectively preserve contrast of small-scale structures for highly accelerated multicontrast volumetric imaging in CNR-limited scenarios. CP-LASER synergistically integrates parallel imaging, low-rank and spatially varying edge-preserving sparse modeling to achieve high signal-to-noise-ratio efficiency by leveraging prior knowledge about signal properties including coil sensitivity weighting, spatiotemporally correlated signal relaxation, and spatially varying sparsity., Results: In the preclinical study using highly accelerated multicontrast volumetric imaging with an isotropic 1.5-mm resolution, CP-LASER demonstrated robust multicontrast reconstruction of peri-infarct characteristics with excellent correspondence with histopathology. CP-LASER provides better delineation of the peri-infarct border zone with improved sharpness than alternative methods in a clinical demonstration on 1.5T with an isotropic 2.2-mm resolution achieved in a single breath-hold., Conclusion: Accelerated multicontrast volumetric imaging with isotropic resolution using CP-LASER has demonstrated the potential to improve peri-infarct characterization in a clinical setting. Magn Reson Med 79:3018-3031, 2018. © 2017 International Society for Magnetic Resonance in Medicine., (© 2017 International Society for Magnetic Resonance in Medicine.)

Background: Previously, a theoretical model based on microvascular physiology was established to facilitate the interpretation of calf perfusion dynamics recorded by arterial spin labeling (ASL)., Purpose: To investigate the clinical relevance of novel perfusion indices by comparing them to the symptoms, response to revascularization, and short-term functional outcome in patients with peripheral arterial disease (PAD)., Study Type: Prospective cohort study., Population: Nineteen patients with PAD., Field Strength/sequence: Pulsed ASL at 3T., Assessment: The mid-calf reactive hyperemia induced by 2 minutes of arterial occlusion was recorded in PAD patients. The perfusion responses were characterized by the peak, time-to-peak, and physiological model-derived indices including the baseline perfusion f r , arterial resistance R a , and compliance C a , and sensitivity g ATP and response time τ ATP of downstream microvasculature to metabolic stress. These indices were compared to the disease severity and outcome within 6 months after revascularization assessed by self-reported symptoms and the ankle-brachial index. Disease severity was categorized as asymptomatic, claudication, or critical limb ischemia. The outcome was categorized as symptom resolved or limited improvement., Statistical Tests: Severity and outcome groups were compared using Mann-Whitney and Kruskal-Wallis tests with Holm-Sidak adjustments., Results: The peak perfusion decreased and model arterial resistance increased progressively with increasing severity of limb ischemia (P = 0.0402 and 0.0413, respectively). Eleven patients had a successful endovascular procedure, including six patients who had symptoms resolved, four patients who had remaining leg pain, and one patient lost to follow-up. The subjects with limited improvement had significantly lower preintervention microvascular sensitivity g ATP than those with symptoms resolved (8.72 ± 1.46 vs. 4.93 ± 0.91, P = 0.0466)., Data Conclusion: ASL reactive hyperemia reflects multiple aspects of the pathophysiology. Measures of macrovascular arterial disease are related to the manifested symptom severity, whereas preintervention g ATP associated with microvascular dysfunction is related to prognosis following revascularization., Level of Evidence: 1 Technical Efficacy: Stage 4 J. Magn. Reson. Imaging 2018;47:1578-1588., (© 2017 International Society for Magnetic Resonance in Medicine.)

Animals detect changes in the environment using modality-specific, peripheral sensory neurons. The insect gustatory system encodes tastant identity and concentration through the independent firing of gustatory receptor neurons (GRNs) that spike rapidly at stimulus onset and quickly adapt. Here, we show the first evidence that concentrated sugar evokes a temporally structured burst pattern of spiking involving two GRNs within the gustatory sensilla of bumblebees. Bursts of spikes resulted when a sucrose-activated GRN was inhibited by another GRN at a frequency of ∼22 Hz during the first 1 s of stimulation. Pharmacological blockade of gap junctions abolished bursting, indicating that bee GRNs have electrical synapses that produce a temporal pattern of spikes when one GRN is activated by a sugar ligand. Bursting permitted bee GRNs to maintain a high rate of spiking and to exhibit the slowest rate of adaptation of any insect species. Feeding bout duration correlated with coherent bursting; only sugar concentrations that produced bursting evoked the bumblebee's feeding reflex. Volume of solution imbibed was a direct function of time in contact with food. We propose that gap junctions among GRNs enable a sustained rate of GRN spiking that is necessary to drive continuous feeding by the bee proboscis., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Magnetic resonance imaging (MRI) has advanced significantly in the past decade and provides a safe and non-invasive method of evaluating peripheral artery disease (PAD), with and without using exogenous contrast agents. MRI offers a promising alternative for imaging patients but the complexity of MRI can make it less accessible for physicians to understand or use. This article provides a brief introduction to the technical principles of MRI for physicians who manage PAD patients. We discuss the basic principles of how MRI works and tailor the discussion to how MRI can evaluate anatomic characteristics of peripheral arterial lesions.

One of the most important tasks of the brain is to learn and remember information associated with food. Studies in mice and Drosophila have shown that sugar rewards must be metabolisable to form lasting memories, but few other animals have been studied. Here, we trained adult, worker honeybees (Apis mellifera) in two olfactory tasks (massed and spaced conditioning) known to affect memory formation to test how the schedule of reinforcement and the nature of a sugar reward affected learning and memory. The antennae and mouthparts of honeybees were most sensitive to sucrose but glucose and fructose were equally phagostimulatory. Whether or not bees could learn the tasks depended on sugar identity and concentration. However, only bees rewarded with glucose or sucrose formed robust long-term memory. This was true for bees trained in both the massed and spaced conditioning tasks. Honeybees fed with glucose or fructose exhibited a surge in haemolymph sugar of greater than 120mM within 30s that remained elevated for as long as 20min after a single feeding event. For bees fed with sucrose, this change in haemolymph glucose and fructose occurred with a 30s delay. Our data showed that olfactory learning in honeybees was affected by sugar identity and concentration, but that olfactory memory was most strongly affected by sugar identity. Taken together, these data suggest that the neural mechanisms involved in memory formation sense rapid changes in haemolymph glucose that occur during and after conditioning., (Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Purpose: Myocardial infarct (MI) may consist of an infarct core (IC) and a heterogeneous, semi-viable border zone (BZ). Patients with chronic MI in the left ventricular (LV) myocardium are at increased risk of developing ventricular arrhythmias, and may therefore qualify for implantable cardioverter defibrillator (ICD) therapy. Indices based on MI mass, as determined by cardiac magnetic resonance (CMR) imaging, are shown to be sensitive in predicting adverse ventricular arrhythmic events. However, several factors, such as imaging technique and spatial resolution affect the accuracy of MI mass quantification. The aim of this study was to compare the MI masses determined by T1-mapping CMR techniques to those of conventional late Gadolinium-enhanced cardiac magnetic resonance (LGE-CMR) using inversion recovery fast gradient echo (IR-FGRE). We additionally aimed to investigate the effect of diminishing image resolution on quantification of the MI mass and its ability to predict appropriate ICD therapy., Methods: Thirty-eight patients with known MI underwent acquisitions of three CMR imaging techniques: the multicontrast late enhancement (MCLE) and modified look-locker inversion recovery (MOLLI) T1-mapping techniques, and conventional inversion recovery fast gradient echo (IR-FGRE) about 20 min after double-dose injection of Gadolinium. We postprocessed images to quantify IC and BZ masses determined by each CMR technique using a full-width half-maximum (FWHM) approach in IR-FGRE images and a fuzzy c-means clustering algorithm for T1-mapping images. To determine the impact of spatial resolution in sensitivity of predicting ICD events, we artificially diminished resolution of MCLE images acquired from a separate group of 27 patients who had been followed up for ICD therapy and compared the MI masses estimated from the original and downsampled MCLE images., Results: Twelve patients out of 27 (44%) received ICD therapy (i.e., one or more delivered shock) during the follow-up stage. Between each of the three imaging methods, IC masses were not significantly different. Conversely, BZ masses determined by MOLLI were larger compared to those determined by MCLE and IR-FGRE (P value = 0.0022 and 0.0003, respectively). The BZ masses determined by MCLE were not significantly different from those determined by IR-FGRE; however, BZ masses determined by the downsampled MCLE were significantly larger than those determined by IR-FGRE and original MCLE (P value = 0.0033 and 0.0003, respectively). The BZ mass estimated by original MCLE was larger in patients who had received ICD therapy compared to those who did not (P value = 0.044). However, when the spatial resolution of the MCLE images was diminished to that of MOLLI, BZ masses were not significantly different between patients with and without ICD therapy., Conclusions: While estimated IC masses were consistent among all three techniques, the estimated BZ masses were not consistent, especially when spatial resolution of images differed between the techniques. In particular, our study showed that diminished image resolution caused an increase in estimation of the BZ mass, likely due to partial volume effects, which led to a reduced sensitivity in the prediction of appropriate ICD therapy., (© 2018 American Association of Physicists in Medicine.)

Background: Radiofrequency (RF) ablation has become a mainstay of treatment for ventricular tachycardia, yet adequate lesion formation remains challenging. This study aims to comprehensively describe the composition and evolution of acute left ventricular (LV) lesions using native-contrast cardiovascular magnetic resonance (CMR) during CMR-guided ablation procedures., Methods: RF ablation was performed using an actively-tracked CMR-enabled catheter guided into the LV of 12 healthy swine to create 14 RF ablation lesions. T 2 maps were acquired immediately post-ablation to visualize myocardial edema at the ablation sites and T 1 -weighted inversion recovery prepared balanced steady-state free precession (IR-SSFP) imaging was used to visualize the lesions. These sequences were repeated concurrently to assess the physiological response following ablation for up to approximately 3 h. Multi-contrast late enhancement (MCLE) imaging was performed to confirm the final pattern of ablation, which was then validated using gross pathology and histology., Results: Edema at the ablation site was detected in T 2 maps acquired as early as 3 min post-ablation. Acute T 2 -derived edematous regions consistently encompassed the T 1 -derived lesions, and expanded significantly throughout the 3-h period post-ablation to 1.7 ± 0.2 times their baseline volumes (mean ± SE, estimated using a linear mixed model determined from n = 13 lesions). T 1 -derived lesions remained approximately stable in volume throughout the same time frame, decreasing to 0.9 ± 0.1 times the baseline volume (mean ± SE, estimated using a linear mixed model, n = 9 lesions)., Conclusions: Combining native T 1 - and T 2 -based imaging showed that distinctive regions of ablation injury are reflected by these contrast mechanisms, and these regions evolve separately throughout the time period of an intervention. An integrated description of the T 1 -derived lesion and T 2 -derived edema provides a detailed picture of acute lesion composition that would be most clinically useful during an ablation case.

Floral nectar is a reward offered by flowering plants to visiting pollinators. Nectar chemistry is important for understanding plant nutrient allocation and plant-pollinator interactions. However, many plant species are difficult to sample as their flowers are small and produce low amounts of nectar.We compared the effects of different methods of nectar collection on the amino acid composition of flowers with low volumes of nectar. We used five methods to collect nectar from 60 (5 × 12) Calluna vulgaris flowers: microcapillary tubes, a low-volume flower rinse (the micro-rinse method, using 2 μl water), filter paper, a high-volume flower rinse (2 ml water) and a flower wash (2 ml water). We analysed the samples for free amino acids using quantitative UHPLC methods .We found that the micro-rinse method (rinsing the nectary with enough water to only cover the nectary) recovered amino acid proportions similar to raw nectar extracted using microcapillary tubes. The filter paper, 2 ml rinse and 2 ml wash methods measured significantly higher values of free amino acids and also altered the profile of amino acids. We discuss our concerns about the increased contamination risk of the filter paper and high-volume rinse and wash samples from dried nectar across the floral tissue (nectar unavailable to floral visitors), pollen, vascular fluid and cellular fluid.Our study will enable researchers to make informed decisions about nectar collection methods depending on their intended chemical analysis. These methods of sampling will enable researchers to examine a larger array of plant species' flowers to include those with low volumes of nectar.

Pollen provides floral visitors with essential nutrients including proteins, lipids, vitamins and minerals. As an important nutrient resource for pollinators, including honeybees and bumblebees, pollen quality is of growing interest in assessing available nutrition to foraging bees. To date, quantifying the protein-bound amino acids in pollen has been difficult and methods rely on large amounts of pollen, typically more than 1 g. More usual is to estimate a crude protein value based on the nitrogen content of pollen, however, such methods provide no information on the distribution of essential and non-essential amino acids constituting the proteins.Here, we describe a method of microwave-assisted acid hydrolysis using low amounts of pollen that allows exploration of amino acid composition, quantified using ultra high performance liquid chromatography (UHPLC), and a back calculation to estimate the crude protein content of pollen.Reliable analysis of protein-bound and free amino acids as well as an estimation of crude protein concentration was obtained from pollen samples as low as 1 mg. Greater variation in both protein-bound and free amino acids was found in pollen sample sizes <1 mg. Due to the variability in recovery of amino acids in smaller sample sizes, we suggest a correction factor to apply to specific sample sizes of pollen in order to estimate total crude protein content.The method described in this paper will allow researchers to explore the composition of amino acids in pollen and will aid research assessing the available nutrition to pollinating animals. This method will be particularly useful in assaying the pollen of wild plants, from which it is difficult to obtain large sample weights.

Objective: To determine the ability of an intraoperative cell salvage (IOCS) system and a leukocyte reduction filter (LRF) to remove hemangiosarcoma (HSA) cells from canine blood., Study Design: Cultured HSA cells were added to canine blood to simulate intraoperative hemorrhage and address hemoabdomen from ruptured splenic HSA. The blood/HSA cell mixture was processed through an IOCS, followed by LRF processing., Sample Population: Whole blood from 3 healthy dogs combined with cultured HSA cells., Methods: The ability of quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), multiparameter flow cytometry, and cytologic examination to detect 50 HSA cells per milliliter of culture media was confirmed. RT-PCR, multiparameter flow cytometry, and cytologic examination were used to determine the presence of cultured HSA cells at 4 points during processing., Results: HSA cells were found in all control samples and in all samples after IOCS but prior to LRF processing with all 3 cell detection methods. HSA cells were not found after IOCS/LRF processing with all 3 cell detection methods., Conclusion: IOCS combined with LRF processing is able to remove cultured HSA cells from canine blood. The addition of LRF to IOCS may allow application of IOCS in dogs with HSA., Clinical Significance: A combination of IOCS and LRF processing may provide an alternative to allogeneic blood transfusion in dogs with hemoabdomen due to HSA., (© 2017 The American College of Veterinary Surgeons.)

Honey bees feed on floral nectar and pollen that they store in their colonies as honey and bee bread. Social division of labor enables the collection of stores of food that are consumed by within-hive bees that convert stored pollen and honey into royal jelly. Royal jelly and other glandular secretions are the primary food of growing larvae and of the queen but are also fed to other colony members. Research clearly shows that bees regulate their intake, like other animals, around specific proportions of macronutrients. This form of regulation is done as individuals and at the colony level by foragers.

Sufficiently diverse and abundant resources are essential for generalist consumers, and form an important part of a suite of conservation strategies for pollinators. Honey bees are generalist foragers and are dependent on diverse forage to adequately meet their nutritional needs. Through analysis of stored pollen (bee bread) samples obtained from 26 honey bee (Apis mellifera L.) hives across NW-England, we quantified bee bread nutritional content and the plant species that produced these stores from pollen. Protein was the most abundant nutrient by mass (63%), followed by carbohydrates (26%). Protein and lipid content (but not carbohydrate) contributed significantly to ordinations of floral diversity, linking dietary quality with forage composition. DNA sequencing of the ITS2 region of the nuclear ribosomal DNA gene identified pollen from 89 distinct plant genera, with each bee bread sample containing between 6 and 35 pollen types. Dominant genera included dandelion (Taraxacum), which was positively correlated with bee bread protein content, and cherry (Prunus), which was negatively correlated with the amount of protein. In addition, proportions of amino acids (e.g. histidine and valine) varied as a function of floral species composition. These results also quantify the effects of individual plant genera on the nutrition of honey bees. We conclude that pollens of different plants act synergistically to influence host nutrition; the pollen diversity of bee bread is linked to its nutrient content. Diverse environments compensate for the loss of individual forage plants, and diversity loss may, therefore, destabilize consumer communities due to restricted access to alternative resources.

Cardiac magnetic resonance imaging (MRI) is appealing to guide complex cardiac procedures because it is ionizing radiation-free and offers flexible soft-tissue contrast. Interventional cardiac MR promises to improve existing procedures and enable new ones for complex arrhythmias, as well as congenital and structural heart disease. Guiding invasive procedures demands faster image acquisition, reconstruction and analysis, as well as intuitive intraprocedural display of imaging data. Standard cardiac MR techniques such as 3D anatomical imaging, cardiac function and flow, parameter mapping, and late-gadolinium enhancement can be used to gather valuable clinical data at various procedural stages. Rapid intraprocedural image analysis can extract and highlight critical information about interventional targets and outcomes. In some cases, real-time interactive imaging is used to provide a continuous stream of images displayed to interventionalists for dynamic device navigation. Alternatively, devices are navigated relative to a roadmap of major cardiac structures generated through fast segmentation and registration. Interventional devices can be visualized and tracked throughout a procedure with specialized imaging methods. In a clinical setting, advanced imaging must be integrated with other clinical tools and patient data. In order to perform these complex procedures, interventional cardiac MR relies on customized equipment, such as interactive imaging environments, in-room image display, audio communication, hemodynamic monitoring and recording systems, and electroanatomical mapping and ablation systems. Operating in this sophisticated environment requires coordination and planning. This review provides an overview of the imaging technology used in MRI-guided cardiac interventions. Specifically, this review outlines clinical targets, standard image acquisition and analysis tools, and the integration of these tools into clinical workflow., Level of Evidence: 1 Technical Efficacy: Stage 5 J. Magn. Reson. Imaging 2017;46:935-950., (© 2017 International Society for Magnetic Resonance in Medicine.)

To characterize and interpret arterial spin labeling (ASL) reactive hyperemia of calf muscles for a better understanding of the microcirculation in peripheral arterial disease (PAD), we present a physiological model incorporating oxygen transport, tissue metabolism, and vascular regulation mechanisms. The model demonstrated distinct effects between arterial stenoses and microvascular dysfunction on reactive hyperemia, and indicated a higher sensitivity of 2-minute thigh cuffing to microvascular dysfunction than 5-minute cuffing. The recorded perfusion responses in PAD patients (n = 9) were better differentiated from the normal subjects (n = 7) using the model-based analysis rather than characterization using the apparent peak and time-to-peak of the responses. The analysis results suggested different amounts of microvascular disease within the patient group. Overall, this work demonstrates a novel analysis method and facilitates understanding of the physiology involved in ASL reactive hyperemia. ASL reactive hyperemia with model-based analysis may be used as a noninvasive microvascular assessment in the presence of arterial stenoses, allowing us to look beyond the macrovascular disease in PAD. A subgroup who will have a poor prognosis after revascularization in the patients with critical limb ischemia may be associated with more severe microvascular diseases, which may potentially be identified using ASL reactive hyperemia.

Toxic nectar is an ecological paradox [1, 2]. Plants divert substantial resources to produce nectar that attracts pollinators [3], but toxins in this reward could disrupt the mutualism and reduce plant fitness [4]. Alternatively, such compounds could protect nectar from robbers [2], provided that they do not significantly alter pollinator visitation to the detriment of plant fitness [1, 5-8]. Indeed, very few studies have investigated the role of plant toxins in nectar for defense against nectar robbers [4, 9, 10]. Here, we compared two Aconitum species (A. napellus and A. lycoctonum) that have flowers specialized for long-tongued bumblebee pollinators (Bombus hortorum) but are occasionally robbed by short-tongued bumblebees (B. terrestris) [6, 11-13]. Pollinator visits to flowers were much more frequent than by robbers, but visits correlated negatively with nectar alkaloid concentration and declined sharply between 200 and 380 ppm. However, alkaloid concentrations of >20 ppm were deterrent to B. terrestris, suggesting that robbers were less tolerant of nectar alkaloids. Nectar of both plant species contained similar concentrations of carbohydrates and toxic alkaloids, but A. lycoctonum was more likely to secrete nectar in each flower and was also visited more frequently by pollinators and robbers. We conclude that alkaloids in Aconitum spp. nectar affect rates of both pollinator visitation and robbery but may have co-evolved with nectar availability to maintain the fitness benefits of specialized plant-pollinator relationships. Chemical defense of nectar is, however, ultimately constrained by pollinator gustatory sensitivity., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Oilseed rape (OSR; Brassica napus L.) is a major crop in temperate regions and provides an important source of nutrition to many of the yield-enhancing insect flower visitors that consume floral nectar. The manipulation of mechanisms that control various crop plant traits for the benefit of pollinators has been suggested in the bid to increase food security, but little is known about inherent floral trait expression in contemporary OSR varieties or the breeding systems used in OSR breeding programmes. We studied a range of floral traits in glasshouse-grown, certified conventional varieties of winter OSR to test for variation among and within breeding systems. We measured 24-h nectar secretion rate, amount, concentration and ratio of nectar sugars per flower, and sizes and number of flowers produced per plant from 24 varieties of OSR representing open-pollinated (OP), genic male sterility (GMS) hybrid and cytoplasmic male sterility (CMS) hybrid breeding systems. Sugar concentration was consistent among and within the breeding systems; however, GMS hybrids produced more nectar and more sugar per flower than CMS hybrid or OP varieties. With the exception of ratio of fructose/glucose in OP varieties, we found that nectar traits were consistent within all the breeding systems. When scaled, GMS hybrids produced 1.73 times more nectar resource per plant than OP varieties. Nectar production and amount of nectar sugar in OSR plants were independent of number and size of flowers. Our data show that floral traits of glasshouse-grown OSR differed among breeding systems, suggesting that manipulation and enhancement of nectar rewards for insect flower visitors, including pollinators, could be included in future OSR breeding programmes.

Purpose: To enable robust reconstruction for highly accelerated three-dimensional multicontrast late enhancement imaging to provide improved MR characterization of myocardial infarction with isotropic high spatial resolution., Theory and Methods: A new method using compressed sensing with low rank and spatially varying edge-preserving constraints (CS-LASER) is proposed to improve the reconstruction of fine image details from highly undersampled data. CS-LASER leverages the low rank feature of the multicontrast volume series in MR relaxation and integrates spatially varying edge preservation into the explicit low rank constrained compressed sensing framework using weighted total variation. With an orthogonal temporal basis pre-estimated, a multiscale iterative reconstruction framework is proposed to enable the practice of CS-LASER with spatially varying weights of appropriate accuracy., Results: In in vivo pig studies with both retrospective and prospective undersamplings, CS-LASER preserved fine image details better and presented tissue characteristics with a higher degree of consistency with histopathology, particularly in the peri-infarct region, than an alternative technique for different acceleration rates. An isotropic resolution of 1.5 mm was achieved in vivo within a single breath-hold using the proposed techniques., Conclusion: Accelerated three-dimensional multicontrast late enhancement with CS-LASER can achieve improved MR characterization of myocardial infarction with high spatial resolution. Magn Reson Med 78:598-610, 2017. © 2016 International Society for Magnetic Resonance in Medicine., (© 2016 International Society for Magnetic Resonance in Medicine.)

Background: The optimal initial noninvasive diagnostic testing strategy for stable coronary artery disease (CAD) is unknown. Although American guidelines recommend an exercise stress test as the first-line test, European guidelines suggest that stress imaging (myocardial perfusion imaging or stress echocardiography) or coronary computed tomography angiography may be preferable. Understanding the relationship between the initial strategy and downstream yield of obstructive CAD and major adverse cardiac events may provide insight as to the optimal strategy., Methods and Results: We conducted a population-based retrospective cohort study of adults in Ontario, Canada, using health administrative and clinical data. The relationship between the initial testing strategy and obstructive CAD on invasive angiography was examined. Patients were then followed from their angiogram onward to determine whether they developed a composite end point of major adverse cardiac events. After adjusting for covariates, patients with initial myocardial perfusion imaging (odds ratio: 0.92; 95% confidence interval, 0.85, 1.00), coronary computed tomography angiography (odds ratio: 1.51; 95% confidence interval, 0.91, 2.49), or stress echo (odds ratio: 0.95; 95% confidence interval, 0.84, 1.08) did not a have significantly different yield of obstructive CAD compared with those with an initial exercise stress test. Furthermore, there was no significant difference in downstream major adverse cardiac events after invasive angiography among the 4 initial testing strategies after adjusting for clinically relevant covariates., Conclusions: Our study found no evidence to suggest significant differences in either yield of obstructive CAD or downstream major adverse cardiac events in patients undergoing an initial noninvasive testing strategy with stress or anatomical imaging compared with those undergoing an initial exercise stress test., (© 2017 The Authors. Published on behalf of the American Heart Association, Inc., by Wiley.)

Background: Myocardial hemorrhage is a frequent complication following reperfusion in acute myocardial infarction and is predictive of adverse outcomes. However, it remains unsettled whether hemorrhage is simply a marker of a severe initial ischemic insult or directly contributes to downstream myocardial damage. Our objective was to evaluate the contribution of hemorrhage towards inflammation, microvascular obstruction and infarct size in a novel porcine model of hemorrhagic myocardial infarction using cardiovascular magnetic resonance (CMR)., Methods: Myocardial hemorrhage was induced via direct intracoronary injection of collagenase in a novel porcine model of ischemic injury. Animals (N = 27) were subjected to coronary balloon occlusion followed by reperfusion and divided into three groups (N = 9/group): 8 min ischemia with collagenase (+HEM); 45 min infarction with saline (I-HEM); and 45 min infarction with collagenase (I+HEM). Comprehensive CMR was performed on a 3 T scanner at baseline and 24 h post-intervention. Cardiac function was quantified by cine imaging, edema/inflammation by T2 mapping, hemorrhage by T2* mapping and infarct/microvascular obstruction size by gadolinium enhancement. Animals were subsequently sacrificed and explanted hearts underwent histopathological assessment for ischemic damage and inflammation., Results: At 24 h, the +HEM group induced only hemorrhage, the I-HEM group resulted in a non-hemorrhagic infarction, and the I+HEM group resulted in infarction and hemorrhage. Notably, the I+HEM group demonstrated greater hemorrhage and edema, larger infarct size and higher incidence of microvascular obstruction. Interestingly, hemorrhage alone (+HEM) also resulted in an observable inflammatory response, similar to that arising from a mild ischemic insult (I-HEM). CMR findings were in good agreement with histological staining patterns., Conclusions: Hemorrhage is not simply a bystander, but an active modulator of tissue response, including inflammation and microvascular and myocardial damage beyond the initial ischemic insult. A mechanistic understanding of the pathophysiology of reperfusion hemorrhage will potentially aid better management of high-risk patients who are prone to adverse long-term outcomes.

Gustatory receptors (Grs) expressed in insect taste neurons signal the presence of carbohydrates, sugar alcohols, CO2, bitter compounds and oviposition stimulants. The honeybee (Apis mellifera) has one of the smallest Gr gene sets (12 Gr genes) of any insect whose genome has been sequenced. Honeybees live in eusocial colonies with a division of labour and perform age-dependent behavioural tasks, primarily food collection. Here, we used RT-qPCR to quantify Gr mRNA in honeybees at two ages (newly-emerged and foraging-age adults) to examine the relationship between age-related physiology and expression of Gr genes. We measured the Gr mRNAs in the taste organs and also the brain and gut. The mRNA of all Gr genes was detected in all tissues analysed but showed plasticity in relative expression across tissues and in relation to age. Overall, Gr gene expression was higher in the taste organs than in the internal tissues but did not show an overall age-dependent difference. In contrast Gr gene expression in brain was generally higher in foragers, which may indicate greater reliance on internal nutrient sensing. Expression of the candidate sugar receptors AmGr1, AmGr2 and AmGr3 in forager brain was affected by the types of sugars bees fed on. The levels of expression in the brain were greater for AmGr1 but lower for AmGr2 and AmGr3 when bees were fed with glucose and fructose compared with sucrose. Additionally, AmGr3 mRNA was increased in starved bees compared to bees provided ad libitum sucrose. Thus, expression of these Grs in forager brain reflects both the satiety state of the bee (AmGr3) and the type of sugar on which the bee has fed.

Lockie, RG, Jalilvand, F, Orjalo, AJ, Giuliano, DV, Moreno, MR, and Wright, GA. A methodological report: Adapting the 505 change-of-direction speed test specific to American football. J Strength Cond Res 31(2): 539-547, 2017-The 505 involves a 10-m sprint past a timing gate, followed by a 180° change-of-direction (COD) performed over 5 m. This methodological report investigated an adapted 505 (A505) designed to be football-specific by changing the distances to 10 and 5 yd. Twenty-five high school football players (6 linemen [LM]; 8 quarterbacks, running backs, and linebackers [QB/RB/LB]; 11 receivers and defensive backs [R/DB]) completed the A505 and 40-yd sprint. The difference between A505 and 0 to 10-yd time determined the COD deficit for each leg. In a follow-up session, 10 subjects completed the A505 again and 10 subjects completed the 505. Reliability was analyzed by t-tests to determine between-session differences, typical error (TE), and coefficient of variation. Test usefulness was examined via TE and smallest worthwhile change (SWC) differences. Pearson's correlations calculated relationships between the A505 and 505, and A505 and COD deficit with the 40-yd sprint. A 1-way analysis of variance (p ≤ 0.05) derived between-position differences in the A505 and COD deficit. There were no between-session differences for the A505 (p = 0.45-0.76; intraclass correlation coefficient = 0.84-0.95; TE = 2.03-4.13%). Additionally, the A505 was capable of detecting moderate performance changes (SWC0.5 > TE). The A505 correlated with the 505 and 40-yard sprint (r = 0.58-0.92), suggesting the modified version assessed similar qualities. Receivers and defensive backs were faster than LM in the A505 for both legs, and right-leg COD deficit. Quarterbacks, running backs, and linebackers were faster than LM in the right-leg A505. The A505 is reliable, can detect moderate performance changes, and can discriminate between football position groups.

The proliferation of cardiac diagnostic tests over the past few decades has received substantial attention from policymakers. However, contemporary population-based temporal trends of the utilization of noninvasive cardiac diagnostic tests for coronary artery disease are not known. Our objective was to examine the temporal trends in the utilization of coronary computed tomography angiography (CCTA), myocardial perfusion imaging (MPI), exercise stress testing (GXT), and stress echocardiography between 2008 and 2014. We performed a population-based repeated cross-sectional study of the adult population of Ontario between January 1, 2008 and December 31, 2014. Annual utilization rates of noninvasive cardiac diagnostic tests were computed. For each cardiac testing modality, a negative binomial regression model was used to assess temporal changes in test utilization. GXT and MPI collectively accounted for 88% of all cardiac noninvasive diagnostic tests throughout our study period. Age- and sex-standardized rates of GXT declined from 26.7/1000 adult population to 21.6/1000 adult population (mean annual reduction of 3.4%; P < 0.001). MPI rates declined from 21.1/1000 adult population to 19.5/1000 adult population (mean annual reduction of 1.3%; P < 0.001). Although utilization rates for both CCTA and stress echocardiography increased over time, the combined rate of all available tests decreased from 50.8/1000 adult population to 49.1/1000 adult population (mean annual reduction of 1.1%; P < 0.001). In conclusion, utilization rates for the most prevalent noninvasive cardiac diagnostic tests-GXT and MPI-declined over our study period. Furthermore, the overall test utilization rate also declined over time. We believe our findings are encouraging from a health policy perspective. Nonetheless, rising utilization rates for CCTA and stress echocardiography will need to be monitored in the future., (Copyright © 2016 Canadian Cardiovascular Society. Published by Elsevier Inc. All rights reserved.)

Objective: MRI in the area of cardiovascular catheter-based interventional procedures is an active field. A common intervention-revascularization of chronic total occlusions, requires a conductive guidewire for revascularization. The mechanical properties of guidewires are paramount to the successful execution of such procedures. Furthermore to benefit from MRI techniques, additional conductors are required to transmit signal from the tip of a catheter. Long thin conductors in MRI systems pose a safety risk in the form of RF heating due to induced RF currents on the conductors. Unfortunately many existing techniques to mitigate this risk require physical modification of the conductors, inevitably resulting in detrimental mechanical tradeoffs in the guidewire. This manuscript proposes a novel application and miniaturization of an existing device, the floating RF trap. The RF trap couples strongly to any thin conductor passing through the trap lumen inducing significant series impedance. This results in reduction of induced RF currents, and thus, heating., Methods and Results: This study shows theoretical and experimental analysis of induced impedance as well as theoretical reduction in heating due to various distributions of traps along the length of a catheter. Results of measuring induced current and heating in phantom experiments are also presented. Through comparison with commercial simulation packages and results of phantom experiments, it is shown that miniaturized RF traps can be modeled accurately, including their induced series impedance and effect on induced RF current., Conclusion and Significance: It was demonstrated that floating RF traps present a feasible method to mitigate RF heating while maintaining important mechanical properties of guidewires.

Purpose: To test and validate magnetic resonance imaging (MRI) sequences for peripheral artery lesion characterization and relate the MRI characteristics to the amount of force required for a guidewire to puncture peripheral chronic total occlusions (CTOs) as a surrogate for immediate failure of endovascular therapy., Methods: Diseased superficial femoral, popliteal, and tibial artery segments containing 55 atherosclerotic lesions were excised from the amputated limbs of 7 patients with critical limb ischemia. The lesions were imaged at high resolution (75 μm 3 voxels) with T2-weighted (T2W) and ultrashort echo time (UTE) sequences on a 7-T MR scanner. The MR images (n=15) were validated with micro-computed tomography and histology. CTOs (n=40) were classified by their MR signal characteristics as "soft" (signals indicating fat, thrombus, microchannels, or loose fibrous tissue), "hard" (collagen and/or speckled calcium signals), or "calcified" (calcified nodule signals). A 2-kg load cell advanced the back end of a 0.035-inch stiff guidewire at a fixed displacement rate (0.05 mm/s) through the CTOs, and the forces required to cross each lesion were measured., Results: T2W images showed fat as hyperintense and hardened tissue as hypointense. Calcium and thrombus appeared as a signal void in conventional MRI sequences but were easily identified in UTE images (thrombus was hyperintense and calcium hypointense). MRI accurately differentiated "hard," "soft," and "calcified" CTOs based on associated guidewire puncture force. The guidewire could not enter "calcified" CTOs (n=6) at all. "Hard" CTOs (n=9) required a significantly higher (p<0.001) puncture force of 1.71±0.51 N vs 0.43±0.36 N for "soft" CTOs (n=25)., Conclusion: MRI characteristics of PAD lesions correlate with guidewire puncture forces, an important aspect of crossability. Future work will determine if clinical MR scanners can be used to predict success in peripheral vascular interventions.

Calcification is a major factor that limits the durability of bioprosthetic valve. A novel bovine pericardial tissue treated with aldehyde capping chemistry and glycerolization was evaluated for its resistance to calcification in comparison with porcine tissues treated with amino oleic acid and bovine pericardial tissue with ethanol rinsing in a rabbit intramuscular model. Tissue discs from the test and control tissues were implanted in rabbits for 60 days. The explanted discs were subject to X-ray imaging, calcium quantification and histology analysis. The test tissue showed 95 and 96 % reduction in calcification in comparison with amino oleic acid treatment and ethanol rinsing treatment, respectively. In addition, the test tissue showed the least inflammatory response as evidenced by a reduced amount of macrophages and giant cells in histology analysis. Furthermore, the aldehyde analysis of the pre-implanted samples showed associated reduction in free aldehyde levels with the test tissue. The reduction in calcification is consistent with previously reported results and is hypothesized to be attributed to the capping of free aldehydes in the test tissue., Competing Interests: This study was sponsored by Edwards Lifesciences LLC. All the authors are Edwards employees and receive salary from the company.

The effect of environmental pollutants on honeybee behaviour has focused mainly on currently used pesticides. However, honeybees are also exposed to persistent organic pollutants (POPs). The aim of this laboratory based study was to determine if exposure to sublethal field-relevant concentrations of POPs altered the spontaneous behaviour of foraging-age worker honeybees. Honeybees (Apis mellifera) were orally exposed to either a sublethal concentration of the polychlorinated biphenyl (PCB) mixture Aroclor 1254 (100 ng/ml), the organochlorine insecticide lindane (2.91 ng/ml) or vehicle (0.01% DMSO, 0.00015% ethanol in 1M sucrose) for 1-4 days. The frequency of single event behaviours and the time engaged in one of four behavioural states (walking, flying, upside down and stationary) were monitored for 15 min after 1, 2, 3 and 4 days exposure. Exposure to Aroclor 1254 but not lindane increased the frequency and time engaged in honeybee motor activity behaviours in comparison to vehicle. The Aroclor 1254-induced hyperactivity was evident after 1 day of exposure and persisted with repeated daily exposure. In contrast, 1 day of exposure to lindane elicited abdominal spasms and increased the frequency of grooming behaviours in comparison to vehicle exposure. After 4 days of exposure, abdominal spasms and increased grooming behaviours were also evident in honeybees exposed to Aroclor 1254. These data demonstrate that POPs can induce distinct behavioural patterns, indicating different toxicokinetic and toxicodynamic properties. The changes in spontaneous behaviour, particularly the PCB-induced chronic hyperactivity and the associated energy demands, may have implications for colony health., Competing Interests: Compliance with Ethical Standards All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed were in accordance with the ethical standards of Newcastle University, UK. This article does not contain any studies with human participants performed by any of the authors. Conflict of Interest The authors declare that they have no conflict of interest.

Bee population declines are linked to the reduction of nutritional resources due to land-use intensification, yet we know little about the specific nutritional needs of many bee species. Pollen provides bees with their primary source of protein and lipids, but nutritional quality varies widely among host-plant species. Therefore, bees might have adapted to assess resource quality and adjust their foraging behavior to balance nutrition from multiple food sources. We tested the ability of two bumble bee species, Bombus terrestris and Bombus impatiens, to regulate protein and lipid intake. We restricted B. terrestris adults to single synthetic diets varying in protein:lipid ratios (P:L). The bees over-ate protein on low-fat diets and over-ate lipid on high-fat diets to reach their targets of lipid and protein, respectively. The bees survived best on a 10:1 P:L diet; the risk of dying increased as a function of dietary lipid when bees ate diets with lipid contents greater than 5:1 P:L. Hypothesizing that the P:L intake target of adult worker bumble bees was between 25:1 and 5:1, we presented workers from both species with unbalanced but complementary paired diets to determine whether they self-select their diet to reach a specific intake target. Bees consumed similar amounts of proteins and lipids in each treatment and averaged a 14:1 P:L for B. terrestris and 12:1 P:L for B. impatiens These results demonstrate that adult worker bumble bees likely select foods that provide them with a specific ratio of P:L. These P:L intake targets could affect pollen foraging in the field and help explain patterns of host-plant species choice by bumble bees., (© 2016. Published by The Company of Biologists Ltd.)

Rationale: Altered cardiac energetics is known to play an important role in the progression toward heart failure. A noninvasive method for imaging metabolic markers that could be used in longitudinal studies would be useful for understanding therapeutic approaches that target metabolism., Objective: To demonstrate the first hyperpolarized 13 C metabolic magnetic resonance imaging of the human heart., Methods and Results: Four healthy subjects underwent conventional proton cardiac magnetic resonance imaging followed by 13 C imaging and spectroscopic acquisition immediately after intravenous administration of a 0.1 mmol/kg dose of hyperpolarized [1- 13 C]pyruvate. All subjects tolerated the procedure well with no adverse effects reported ≤1 month post procedure. The [1- 13 C]pyruvate signal appeared within the chambers but not within the muscle. Imaging of the downstream metabolites showed 13 C-bicarbonate signal mainly confined to the left ventricular myocardium, whereas the [1- 13 C]lactate signal appeared both within the chambers and in the myocardium. The mean 13 C image signal:noise ratio was 115 for [1- 13 C]pyruvate, 56 for 13 C-bicarbonate, and 53 for [1- 13 C]lactate., Conclusions: These results represent the first 13 C images of the human heart. The appearance of 13 C-bicarbonate signal after administration of hyperpolarized [1- 13 C]pyruvate was readily detected in this healthy cohort (n=4). This shows that assessment of pyruvate metabolism in vivo in humans is feasible using current technology., Clinical Trial Registration: URL: https://www.clinicaltrials.gov. Unique identifier: NCT02648009., (© 2016 The Authors.)