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The transcriptome from lupulin glands and associated bracts from cone tissue of hop (Humulus lupulus) c.v. ‘Cascade’, during three stages of development: early, mid, and late or near-harvest, was s...

COX-2 fluorescent probes are promising tools for cancer diagnosis. Such probes have been conventionally designed by conjugating a fluorophore to COX-2 inhibitors through lengthy synthetic processes. Herein, a type of fluorescent probe for COX-2 imaging has been developed using a single-step process from rofecoxib. In total, six rofecoxib analogues were designed using this unique strategy. Several analogues retained comparative COX-2 targeting activity of rofecoxib and also exhibited attractive fluorescent properties, which were investigated using a combination of experimental and theoretical approaches. The most potent analogue

Hops are valued for their secondary metabolites, including bitter acids, flavonoids, oils, and polyphenols, that impart flavor in beer. Previous studies have shown that hop yield and bitter acid content decline with increased temperatures and low-water stress. We looked at physiological traits and differential gene expression in leaf, stem, and root tissue from hop (Humulus lupulus) cv. USDA Cascade in plants exposed to high temperature stress, low-water stress, and a compound treatment of both high temperature and low-water stress for six weeks. The stress conditions imposed in these experiments caused substantial changes to the transcriptome, with significant reductions in the expression of numerous genes involved in secondary metabolite biosynthesis. Of the genes involved in bitter acid production, the critical gene valerophenone synthase (VPS) experienced significant reductions in expression levels across stress treatments, suggesting stress-induced lability in this gene and/or its regulatory elements may be at least partially responsible for previously reported declines in bitter acid content. We also identified a number of transcripts with homology to genes shown to affect abiotic stress tolerance in other plants that may be useful as markers for breeding improved abiotic stress tolerance in hop. Lastly, we provide the first transcriptome from hop root tissue.

Cerebrovascular dysfunction is a critical risk factor for the pathogenesis of Alzheimer’s disease (AD). The purinergic P2Y2 receptor and endoplasmic reticulum (ER) stress are tightly associated with vascular dysfunction and the pathogenesis of AD. However, the protective effects of exercise training on P2Y2 receptor- and ER stress-associated cerebrovascular dysfunction in AD are mostly unknown. Control (C57BL/6, CON) and AD (APP/PS1dE9, AD) mice underwent treadmill exercise training (EX). 2-MeS-ATP-induced dose-dependent vasoreactivity was determined by using a pressurized posterior cerebral artery (PCA) from 10–12-mo-old mice. Human brain microvascular endothelial cells (HBMECs) were exposed to laminar shear stress (LSS) at 20 dyn/cm(2) for 30 min, 2 h, and 24 h. The expression of P2Y2 receptors, endothelial nitric oxide synthase (eNOS), and ER stress signaling were quantified by Western blot analysis. Notably, exercise converted ATP-induced vasoconstriction in the PCA from AD mice to vasodilation in AD+EX mice to a degree commensurate to the vascular reactivity observed in CON mice. Exercise reduced the expression of amyloid peptide precursor (APP) and increased the P2Y2 receptor and Akt/eNOS expression in AD mice brain. Mechanistically, LSS increased the expression of both P2Y2 receptor and eNOS protein in HBMECs, but these increases were blunted by a P2Y2 receptor antagonist in HBMECs. Exercise also reduced the expression of aberrant ER stress markers p-IRE1, p/t-eIF2α, and CHOP, as well as Bax/Bcl-2, in AD mice brain. Collectively, our results demonstrate for the first time that exercise mitigates cerebrovascular dysfunction in AD through modulating P2Y2 receptor- and ER stress-dependent endothelial dysfunction. NEW & NOTEWORTHY A limited study has investigated whether exercise training can improve cerebrovascular function in Alzheimer's disease. The novel findings of the study are that exercise training improves cerebrovascular dysfunction through enhancing P2Y2 receptor-mediated eNOS signaling and reducing ER stress-associated pathways in AD. These data suggest that exercise training, which regulates P2Y2 receptor and ER stress in AD brain, is a potential therapeutic strategy for Alzheimer's disease.

The abnormal phosphorylation of tau is a necessary precursor to the formation of tau fibrils, a marker of Alzheimer's disease. We hypothesize that hyperphosphorylative conditions may result in unique cell surface markers. We identify and demonstrate the utility of such surrogate markers to identify the hyperphosphorylative state.

The development of imaging agents for in vivo detection of alpha-synuclein (α-syn) pathologies faces several challenges. A major gap in the field is the lack of diverse molecular scaffolds with high affinity and selectivity to α-syn fibrils for in vitro screening assays. Better in vitro scaffolds can instruct the discovery of better in vivo agents. We report the rational design, synthesis, and in vitro evaluation of a series of novel 1-indanone and 1,3-indandione derivatives from a Structure-Activity Relationship (SAR) study centered on some existing α-syn fibril binding ligands. Our results from fibril saturation binding experiments show that two of the lead candidates compounds 8 and 32 bind α-syn fibrils with binding constants (K(d)) of 9.0 and 18.8 nM, respectively, and selectivity of greater than 10x for α-syn fibrils compared with amyloid-β (Aβ) and tau fibrils. Our results demonstrate that the lead ligands avidly label all forms of α-syn on PD brain tissue sections, but only the dense core of senile plaques in AD brain tissue, respectively. These results are corroborated by ligand-antibody colocalization data from Syn211, which shows immunoreactivity towards all forms of α-syn aggregates, and Syn303, which displays preferential reactivity towards mature Lewy pathology. Our results reveal that 1-indanone derivatives have desirable properties for the biological evaluation of α-synucleinopathies.

Large amounts of nitrogen fertilizers applied during lettuce production are lost due to leaching or volatilization, causing severe environmental pollution and increased costs of production. Developing lettuce varieties with high nitrogen use efficiency (NUE) is the eco-friendly solution to reduce nitrogen pollution. Hence, in-depth knowledge of nitrogen metabolism and assimilation genes and their regulation is critical for developing high NUE varieties. In this study, we performed comparative transcriptomic analysis of the cultivated lettuce (Lactuca sativa L.) and its wild progenitor (L. serriola) under high and low nitrogen conditions. A total of 2,704 differentially expressing genes (DEGs) were identified. Key enriched biological processes included photosynthesis, oxidation-reduction process, chlorophyll biosynthetic process, and cell redox homeostasis. The transcription factors (TFs) belonging to the ethylene responsive factor (ERF) family and basic helix-loop-helix (bHLH) family were among the top differentially expressed TFs. Using weighted gene co-expression network analysis (WGCNA) we constructed nine co-expression modules. Among these, two modules were further investigated because of their significant association with total nitrogen content and photosynthetic efficiency of photosystem II (PSII). Three highly correlated clusters were identified which included hub genes for nitrogen metabolism, secondary metabolites, and carbon assimilation, and were regulated by cluster specific TFs. We found that the expression of nitrogen transportation and assimilation genes varied significantly between the two lettuce species thereby providing the opportunity of introgressing wild alleles into the cultivated germplasm for developing lettuce cultivars with more efficient use of nitrogen.

Electrification of endothermic reactions has the potentialto provide a compact and flexible reactor concept, and at the same time, substantially reduce CO2 emissions relative to combustion-heated processes. Here, we show how integrated electrical heating using a wash-coated catalytic structure can resolve limiting thermal conductivity acrossthe catalyst. The inherent uniform supply of heat enables engineeringof catalytic efficiency to desired values by changing wash-coat thickness.Overall, the approach diminishes catalytic efficiency as a limitingdesign parameter. Instead, coat thickness will relate to catalyst lifetime, as very thin coats are more susceptible to deactivation. Characteristic time-scale analysis indicates heat transfer to be theleast limiting mechanism, and reactor performance is instead governed by diffusion. Optimal performance based on fluid dynamic simulations favors internal diameters below 0.5 mm, and high linear gas velocities, toward alleviating mass transfer limitations. Integrated electrical heated steam methane reforming essentially inverts the order of reaction mechanisms compared to conventional reforming, challenging the constraintsof current industrial practice.

More-efficient heating Large-scale production of hydrogen through steam reforming directly produces CO 2 as a side product. In addition, the heating of reactors through fossil-fuel burning contributes further CO 2 emissions. One problem is that the catalyst bed is heated unevenly, which renders much of the catalyst effectively inactive. Wismann et al. describe an electrical heating scheme for a metal tube reactor that improves the uniformity of heating and catalyst usage (see the Perspective by Van Geem et al. ). Adoption of this alternative approach could affect CO 2 emissions by up to approximately 1% of global emissions. Science , this issue p. 756 ; see also p. 734

Advances in high-throughput imaging allow researchers to collect three-dimensional images of whole organ microvascular networks. These extremely large images contain networks that are highly complex, time consuming to segment, and difficult to visualize. In this paper, we present a framework for segmenting and visualizing vascular networks from terabyte-sized three-dimensional images collected using high-throughput microscopy. While these images require terabytes of storage, the volume devoted to the fiber network is $\approx 4$ percent of the total volume size. While the networks themselves are sparse, they are tremendously complex, interconnected, and vary widely in diameter. We describe a parallel GPU-based predictor-corrector method for tracing filaments that is robust to noise and sampling errors common in these data sets. We also propose a number of visualization techniques designed to convey the complex statistical descriptions of fibers across large tissue sections—including commonly studied microvascular characteristics, such as orientation and volume.

Gut-associated lymphoid tissues (GALT) serve as key priming sites for intestinal adaptive immune responses. Most of our understanding of GALT function and development arises from studies in mice. However, the diversity, structure and cellular composition of GALT differs markedly between mammalian species and the developmental window in which distinct GALT structures develop in large mammals remains poorly understood. Given the importance of pigs as models of human disease, as well as their role in livestock production, we adapted a recently developed protocol for the isolation of human GALT to assess the diversity, development and immune composition of large intestinal GALT in neonatal and adult pigs. We demonstrate that the large intestine of adult pigs contains two major GALT types; multifollicular submucosal GALT that we term submucosal lymphoid clusters (SLC) which develop prenatally, and as yet undescribed mucosal isolated lymphoid follicles (M-ILF), which arise after birth. Using confocal laser microscopy and flow cytometry, we additionally assess the microanatomy and lymphocyte composition of SLC and M-ILF, compare them to jejunal Peyer's patches (PP), and describe the maturation of these structures. Collectively, our results provide a deeper understanding of the diversity and development of GALT within the porcine large intestine.

In this study, multiple measures of angiogenic processes were assessed in murine brain endothelial (bEnd.3) cells after exposure to the stable prostacyclin analog, iloprost. Additionally, changes in the γ-secretase enzyme were evaluated after activation of prostacyclin signaling using PGI2overexpressing mouse brain tissue and immunohistology studies in bEnd.3 cells. A three-dimensional assay of tube formation revealed that iloprost inhibits normal formation by significantly reduced tube lengths and vessel mesh area. The iloprost-mediated inhibition of tube-like structures was ameliorated by a specific IP-receptor antagonist, CAY10449. Reductions in wound healing were observed with iloprost application in a dose-dependent manner and this effect was reversed using CAY10449. Iloprost did not exhibit anti-proliferative effects in the bEnd.3 cells. When subjected to a Transwell assay to evaluate changes in trans-epithelial electrical resistance (TEER), bEnd.3 cells displayed reduced TEER values in the presence of iloprost an effect that lasted over prolonged periods (24 hours). Again, CAY10449 was able to reverse iloprost-mediated reductions in TEER value. Surprisingly, the adenylyl cyclase activator, forskolin, produced higher TEER values in the bEnd.3 cells over the same time. The TEER results suggest that iloprost may not activating the Gs protein of the IP receptor to increase cAMP levels given by the opposing results seen with iloprost and forskolin. In terms of γ-secretase expression, PGI2overexpression in mice increased the expression of the APH-1α subunit in the hippocampus and cortex. In bEnd.3 cells, iloprost application slightly increased APH-1α subunit expression measured by western blot and interrupted the colocalization of Presenilin 1 and APH-1α subunits using immunohistochemistry. The results suggest that prostacyclin signaling within bEnd.3 cells is anti-angiogenic and further downstream events have effects on the expression and most likely the activity of the Aβ cleaving enzyme, γ-secretase.

The development of imaging agents for in vivo detection of alpha-synuclein (α-syn) pathologies faces several challenges. A major gap in the field is the lack of diverse molecular scaffolds with high affinity and selectivity to α-syn fibrils for in vitro screening assays. Better in vitro scaffolds can instruct the discovery of better in vivo agents. We report the rational design, synthesis, and in vitro evaluation of a series of novel 1-indanone and 1,3-indandione derivatives from a Structure-Activity Relationship (SAR) study centered on some existing α-syn fibril binding ligands. Our results from fibril saturation binding experiments show that two of the lead candidates bind α-syn fibrils with binding constants (Kd) of 9.0 and 18.8 nM, respectively, and selectivity of greater than 10x for α-syn fibrils compared with amyloid-β (Aβ) fibrils. Our results demonstrate that the lead ligands avidly label all forms of α-syn on PD brain tissue sections, but only the dense core of senile plaques in AD brain tissue, respectively. These results are corroborated by ligand-antibody colocalization data from Syn211, which shows immunoreactivity towards all forms of α-syn aggregates, and Syn303, which displays preferential reactivity towards mature Lewy pathology. Our results reveal that 1-indanone derivatives have desirable properties for the biological evaluation of α-synucleinopathies.

Deep understanding of genetic architecture of water-stress tolerance is critical for efficient and optimal development of water-stress tolerant cultivars, which is the most economical and environmentally sound approach to maintain lettuce production with limited irrigation. Lettuce (Lactuca sativa L.) production in areas with limited precipitation relies heavily on the use of ground water for irrigation. Lettuce plants are highly susceptible to water-stress, which also affects their nutrient uptake efficiency. Water stressed plants show reduced growth, lower biomass, and early bolting and flowering resulting in bitter flavors. Traditional phenotyping methods to evaluate water-stress are labor intensive, time-consuming and prone to errors. High throughput phenotyping platforms using kinetic chlorophyll fluorescence and hyperspectral imaging can effectively attain physiological traits related to photosynthesis and secondary metabolites that can enhance breeding efficiency for water-stress tolerance. Kinetic chlorophyll fluorescence and hyperspectral imaging along with traditional horticultural traits identified genomic loci affected by water-stress. Supervised machine learning models were evaluated for their accuracy to distinguish water-stressed plants and to identify the most important water-stress related parameters in lettuce. Random Forest (RF) had classification accuracy of 89.7% using kinetic chlorophyll fluorescence parameters and Neural Network (NN) had classification accuracy of 89.8% using hyperspectral imaging derived vegetation indices. The top ten chlorophyll fluorescence parameters and vegetation indices selected by sequential forward selection by RF and NN were genetically mapped using a L. sativa × L. serriola interspecific recombinant inbred line (RIL) population. A total of 25 quantitative trait loci (QTL) segregating for water-stress related horticultural traits, 26 QTL for the chlorophyll fluorescence traits and 34 QTL for spectral vegetation indices (VI) were identified. The percent phenotypic variation (PV) explained by the horticultural QTL ranged from 6.41 to 19.5%, PV explained by chlorophyll fluorescence QTL ranged from 6.93 to 13.26% while the PV explained by the VI QTL ranged from 7.2 to 17.19%. Eight QTL clusters harboring co-localized QTL for horticultural traits, chlorophyll fluorescence parameters and VI were identified on six lettuce chromosomes. Molecular markers linked to the mapped QTL clusters can be targeted for marker-assisted selection to develop water-stress tolerant lettuce.

BackgroundLifestyle therapy with resistance training is a potent measure to counteract age-related loss in muscle strength and mass. Unfortunately, many individuals fail to respond in the expected manner. This phenomenon is particularly common among older adults and those chronically diseased (e.g. chronic obstructive pulmonary disease, COPD), and may involve endocrine variables such as vitamin D. At present, the effects of vitamin D supplementation on responses to resistance training remain largely unexplored.MethodsNinety-five male and female participants (healthy, n=71; COPD, n=24; age 68 ± 5 years) were randomly assigned to receive either vitamin D3 or placebo supplementation for 28 weeks in a double-blinded manner (latitude 61°N, September-May). Seventy-eight participants completed the RCT, which was initiated by 12 weeks of supplementation-only (in average, 3333 IU.day-1), followed by 13 weeks of combined supplementation (2000 IU.day-1) and supervised whole-body resistance training (twice weekly), interspersed with testing and measurements. Outcome measures included multiple assessments of muscle strength (n=7), endurance performance (n=6), and muscle mass (n=3, legs, primary), as well as muscle quality (legs), muscle biology (m. vastus lateralis; muscle fibre characteristics, transcriptome), and health-related variables (e.g. visceral fat mass and blood lipid profile). For main outcome domains such as muscle strength and muscle mass, weighted combined factors were calculated from the range of singular assessments.ResultsOverall, 13 weeks of resistance training increased muscle strength (13% ± 8%), muscle mass (9% ± 8%) and endurance performance (one-legged, 23% ± 15%; whole-body, 8% ± 7%), assessed as weighted combined factors, and were associated with changes in health variables (e.g. visceral fat, − 6% ± 21%; [LDL]serum, −4% ± 14%) and muscle tissue characteristics such as fibre type proportions (e.g. IIX, −3%-points), myonuclei·fibre-1 (30% ± 65%), total RNA/rRNA abundances (15%/6-19%), and transcriptome profiles (e.g. ∼336 differentially expressed genes). Vitamin D3 supplementation led to robust increases in [25(OH)D]serum (Δ49% vs placebo), but did not affect training-associated changes for any of the main outcome domains, with no interaction being evident with disease status or pre-RCT [25(OH)D]serum. In secondary analyses, vitamin D3 affected expression of gene sets involved in vascular functions in muscle tissue, strength gains in participants with high fat mass, and [cortisol]serum (Δ20%), all of which advocate further study.ConclusionsVitamin D3 supplementation did not affect muscular responses to resistance training in old adults with or without COPD.

Alzheimer’s disease (AD) is a progressive neurodegenerative disorder that is the most common cause of dementia in aged populations. A substantial amount of data demonstrates that chronic neuroinflammation can accelerate neurodegenerative pathologies, while epidemiological and experimental evidence suggests that the use of anti-inflammatory agents may be neuroprotective. In AD, chronic neuroinflammation results in the upregulation of cyclooxygenase and increased production of prostaglandin H2, a precursor for many vasoactive prostanoids. While it is well-established that many prostaglandins can modulate the progression of neurodegenerative disorders, the role of prostacyclin (PGI2) in the brain is poorly understood. We have conducted studies to assess the effect of elevated prostacyclin biosynthesis in a mouse model of AD. Upregulated prostacyclin expression significantly worsened multiple measures associated with amyloid disease pathologies. Mice overexpressing both amyloid and PGI2exhibited impaired learning and memory and increased anxiety-like behavior compared with non-transgenic and PGI2control mice. PGI2overexpression accelerated the development of amyloid accumulation in the brain and selectively increased the production of soluble amyloid-β 42. PGI2damaged the microvasculature through alterations in vascular length and branching; amyloid expression exacerbated these effects. Our findings demonstrate that chronic prostacyclin expression plays a novel and unexpected role that hastens the development of the AD phenotype.

A comprehensive understanding of microvascular networks is required to generate platforms that track treatment efficacy. Current approaches are labor intensive and limited to small tissue volumes. In this work we describe an acquisition and segmentation framework for low-cost imaging and microvascular modeling at resolution comparable to confocal with data rates comparable to light-sheet microscopy. Segmentation is performed using a GPU-based method that extracts microvascular structure and connectivity embedded in these images. The microvasculature network is stored such that the graph [G = V, E] structure can be exploited to quantify large-scale angiomes and facilitate data mining at the terabyte scale.

Super Resolution (SR) microscopy leverages a variety of optical and computational techniques for overcoming the optical diffraction limit to acquire additional spatial details. However, added spatial details challenge existing segmentation tools. Confounding features include protein distributions that form membranes and boundaries, such as cellular and nuclear surfaces. We present a segmentation pipeline that retains the benefits provided by SR in surface separation while providing a tensor field to overcome these confounding features. The proposed technique leverages perceptual grouping to generate a tensor field that enables robust evolution of active contours despite ill-defined membrane boundaries.

The red alga Porphyra umbilicalis Kutzing has a broad distribution within the North Atlantic. In the Northeast Atlantic, P. umbilicalis is dioecious and reproduces both sexually and asexually, while in the Northwest Atlantic, only asexual reproduction has been observed. In this study, transcriptomes were mined to identify putative single nucleotide polymorphisms (SNP) markers. A computational pipeline was developed that accounts for the specific characteristics of transcriptome dataset, filtered against the available red alga Chondrus genome and P. umbilicalis EST library to eliminate microbial contamination. Five hundred forty-nine putative SNPs were detected within a single population (Schoodic Point, ME, USA). Five of the validated SNP markers were applied in a pilot study of genetic diversity and population structure of seven P. umbilicalis populations within the Gulf of Maine. Results of this study revealed the genetic diversity and structure of P. umbilicalis populations in the Gulf of Maine. Novel genotypes were found in the open coastal populations at Reid State Park, Schoodic Point, and the estuarine tidal rapid population at Wiscasset. Our study represents the first attempt to develop suitable bioinformatic pipeline for RNA-seq to detect SNP markers for red alga Porphyra umbilicalis and successfully used these SNP markers for population study.

Increasing implementation of renewable energy requires an infrastructure compatible with the intermittent production of green electricity. Herein we show the flexibility of electrically heated steam methane reforming with integrated ohmic heating, through a combination of CFD modelling and lab scale reactor tests. It is shown how start-up from an idle state to operation conditions can be achieved with instantaneous application of the full power required for a steady state conversion of 80%, with initial heating rates exceeding 50 °C/min. The initial heating rate is correlated to the thermal mass of the reactor, with the endothermic reaction governing the temperature profile. Cyclical operation displays no apparent delay between the change in temperature and methane conversion. The highest thermal gradient across the washcoat is predicted at steady state, with no increase during start-up despite the higher heating rates. The highest risk of carbon formation is predicted at the inlet at steady state operation. A temporarily peak in the equilibrated carbon potential is predicted near the outlet during start-up and shutdown between 500 and 600 °C, governed by the thermodynamics of the feed composition. Integrated ohmic heating supports steam methane reforming scalable to industrial conditions, operating closer to thermodynamic limits for carbon formation, and potentially based on the access to intermittent excess of renewable energy.

Analysis of three-dimensional biological samples is critical to understanding tissue function and the mechanisms of disease. Many chronic conditions, like neurodegenerative diseases and cancers, correlate with complex tissue changes that are difficult to explore using two-dimensional histology. While three-dimensional techniques such as confocal and light-sheet microscopy are well-established, they are time consuming, require expensive instrumentation, and are limited to small tissue volumes. Three-dimensional microscopy is therefore impractical in clinical settings and often limited to core facilities at major research institutions. There would be a tremendous benefit to providing clinicians and researchers with the ability to routinely image large three-dimensional tissue volumes at cellular resolution. In this paper, we propose an imaging methodology that enables fast and inexpensive three-dimensional imaging that can be readily integrated into current histology pipelines. This method relies on block-face imaging of paraffin-embedded samples using deep-ultraviolet excitation. The imaged surface is then ablated to reveal the next tissue section for imaging. The final image stack is then aligned and reconstructed to provide tissue models that exceed the depth and resolution achievable with modern three-dimensional imaging systems.

Microvessels are frequent targets for research into tissue development and disease progression. These complex and subtle differences between networks are currently difficult to visualize, making sample comparisons subjective and difficult to quantify. These challenges are due to the structure of microvascular networks, which are sparse but space-filling. This results in a complex and interconnected mesh that is difficult to represent and impractical to interpret using conventional visualization techniques. We develop a bi-modal visualization framework, leveraging graph-based and geometry-based techniques to achieve interactive visualization of microvascular networks. This framework allows researchers to objectively interpret the complex and subtle variations that arise when comparing microvascular networks.

This chapter examines the confluence of art and science in a collaborative project between an artist, Janet Biggs, and two scientists, Jason Eriksen and Jokūbas Žiburkus. The project delves into memory loss and Alzheimer’s disease for Biggs’ 2015 exhibition at the Blaffer Museum of Art.