bluuebunny/crossref_metadata_2025 huggingface dataset:
Minimal convex extensions and finite difference discretisation of the quadratic Monge–Kantorovich problem
Authors: BENAMOU JEAN-DAVID , DUVAL VINCENT
Published: September 2018

We present an adaptation of the Monge–Ampère (MA) lattice basis reduction scheme to the MA equation with second boundary value condition, provided the target is a convex set. This yields a fast adaptive method to numerically solve the optimal transport (OT) problem between two absolutely continuous measures, the second of which has convex support. The proposed numerical method actually captures a specific Brenier solution which is minimal in some sense. We prove the convergence of the method as the grid step size vanishes and show with numerical experiments that it is able to reproduce subtle properties of the OT problem.
Mögliche Wege zur Erweiterung interkultureller Perspektiven in der Pädagogik
Authors: Holstein Bettina , Kindervater Angela
Published: September 2018

Zusammenfassung. Es ist notwendig, kultursensible Kompetenzen im Rahmen der Ausbildung von Gesundheitsberufen zu fördern. Nicht nur die seit 2015 steigende Anzahl von Migranten2 verdeutlicht, wie wichtig die Entwicklung kultursensibler Kompetenzen ist. Basierend auf den Ergebnissen einer quantitativen Studie (Holstein, 2016) wird diskutiert, was Unterricht in diesem Bereich leisten sollte und welche Dimensionen die kultursensible Kompetenz impliziert. Möglichkeiten zur Förderung werden vorgestellt und diskutiert.
Getting Your Paper Published Part 2: Good Grammar, Clear Figures
Authors: Robock Alan
Published: September 2018

Recommendations from an AGU journal editor on ensuring that your manuscript is well written and clearly presented.
Fracture Properties of Nash Point Shale as a Function of Orientation to Bedding
Authors: Forbes Inskip Nathaniel D. , Meredith Philip G. , Chandler Michael R. , Gudmundsson Agust
Published: September 2018

Understanding how fracture networks develop in shale formations is important when exploiting unconventional hydrocarbon reservoirs and analyzing the integrity of the seals of conventional and carbon capture and storage reservoirs. Despite this importance, experimentally derived fracture data for shale remains sparse. Here we characterize shale from Nash Point in South Wales, United Kingdom, in terms of ultrasonic wave velocities, tensile strength, and fracture toughness (KIc). We measure these properties in multiple orientations, including angles oblique to the three principal fracture orientations—Short‐transverse, Arrester, and Divider. We find that the Nash Point shale is mechanically highly anisotropic, with tensile strength and KIc values lowest in the Short‐transverse orientation and highest in the Arrester and Divider orientations. Fractures that propagate in a direction oblique or normal to bedding commonly deflect toward the weaker Short‐transverse orientation. Such deflected fractures can no longer be considered to propagate in pure mode‐I. We therefore present a method to correct measured KIc values to account for deflection by calculating mode‐I and mode‐II deflection stress intensities (KId and KIId, respectively). Because of the mixed‐mode nature of deflected fractures, we adopt a fracture (Gc) energy‐based approach that allows analysis of critical fracture propagation conditions for both deflected and undeflected fractures in all orientations. We find that Gc increases as the angle from the Short‐transverse plane increases. We conclude that a modified elliptical function, previously applied to tensile strength and KIc, can be used to estimate values of Gc at angles between the Short‐transverse and Arrester orientations.
Cholinergic basal forebrain neurons regulate fear extinction consolidation through p75 neurotrophin receptor signaling
Authors: Boskovic Zoran , Milne Michael R. , Qian Lei , Clifton Hamish D. , McGovern Alice E. , Turnbull Marion T. , Mazzone Stuart B. , Coulson Elizabeth J.
Published: September 2018

Cholinergic basal forebrain (cBF)-derived neurotransmission plays a crucial role in regulating neuronal function throughout the cortex, yet the mechanisms controlling cholinergic innervation to downstream targets have not been elucidated. Here we report that removing the p75 neurotrophin receptor (p75NTR) from cBF neurons induces a significant impairment in fear extinction consolidation. We demonstrate that this is achieved through alterations in synaptic connectivity and functional activity within the medial prefrontal cortex. These deficits revert back to wild-type levels upon re-expression of the active domain of p75NTRin adult animals. These findings demonstrate a novel role for cholinergic neurons in fear extinction consolidation and suggest that neurotrophic signaling is a key regulator of cholinergic-cortical innervation and function.
Misoprostol regulates Bnip3 repression and alternative splicing to control cellular calcium homeostasis during hypoxic stress
Authors: Field Jared T. , Martens Matthew D. , Mughal Wajihah , Hai Yan , Chapman Donald , Hatch Grant M. , Ivanco Tammy L. , Diehl-Jones William , Gordon Joseph W.
Published: September 2018

The cellular response to hypoxia involves the activation of a conserved pathway for gene expression regulated by the transcription factor complex called hypoxia-inducible factor (HIF). This pathway has been implicated in both the adaptive response to hypoxia and in several hypoxic-ischemic-related pathologies. Perinatal hypoxic injury, often associated with prematurity, leads to multi-organ dysfunction resulting in significant morbidity and mortality. Using a rodent model of neonatal hypoxia and several representative cell lines, we observed HIF1α activation and down-stream induction of the cell death gene Bnip3 in brain, large intestine, and heart which was mitigated by administration of the prostaglandin E1 analog misoprostol. Mechanistically, we determined that misoprostol inhibits full-length Bnip3 (Bnip3-FL) expression through PKA-mediated NF-κB (P65) nuclear retention, and the induction of pro-survival splice variants. We observed that the dominant small pro-survival variant of Bnip3 in mouse cells lacks the third exon (Bnip3ΔExon3), whereas human cells produce a pro-survival BNIP3 variant lacking exon 2 (BNIP3ΔExon2). In addition, these small Bnip3 splice variants prevent mitochondrial dysfunction, permeability transition, and necrosis triggered by Bnip3-FL by blocking calcium transfer from the sarco/endoplasmic reticulum to the mitochondria. Furthermore, misoprostol and Bnip3ΔExon3 promote nuclear calcium accumulation, resulting in HDAC5 nuclear export, NFAT activation, and adaptive changes in cell morphology and gene expression. Collectively, our data suggests that misoprostol can mitigate the potential damaging effects of hypoxia on multiple cell types by activating adaptive cell survival pathways through Bnip3 repression and alternative splicing.
Non-insulin determinant pathways maintain glucose homeostasis upon metabolic surgery
Authors: Lu Zongshi , Wei Xiao , Sun Fang , Zhang Hexuan , Gao Peng , Pu Yunfei , Wang Anlong , Chen Jing , Tong Weidong , Li Qiang , Zhou Xunmei , Yan Zhencheng , Zheng Hongting , Yang Gangyi , Huang Yu , Liu Daoyan , Zhu Zhiming
Published: September 2018

Insulin is critical for glucose homeostasis, and insulin deficiency or resistance leads to the development of diabetes. Recent evidence suggests that diabetes can be remitted independent of insulin. However, the underlying mechanism remains largely elusive. In this study, we utilized metabolic surgery as a tool to identify the non-insulin determinant mechanism. Here, we report that the most common metabolic surgery, Roux-en-Y gastric bypass (RYGB), reduced insulin production but persistently maintained euglycemia in healthy Sprague-Dawley (SD) rats and C57 mice. This reduction in insulin production was associated with RYGB-mediated inhibition of pancreatic preproinsulin and polypyrimidine tract-binding protein 1. In addition, RYGB also weakened insulin sensitivity that was evaluated by hyperinsulinemic-euglycemic clamp test and downregulated signaling pathways in insulin-sensitive tissues. The mechanistic evidence suggests that RYGB predominately shifted the metabolic profile from glucose utilization to fatty acid oxidation, enhanced the energy expenditure and activated multiple metabolic pathways through reducing gut energy uptake. Importantly, the unique effect of RYGB was extended to rats with islet disruption and patients with type 2 diabetes. These results demonstrate that compulsory rearrangement of the gastrointestinal tract can initiate non-insulin determinant pathways to maintain glucose homeostasis. Based on the principle of RYGB action, the development of a noninvasive intervention of the gastrointestinal tract is a promising therapeutic route to combat disorders characterized by energy metabolism dysregulation.
Genetic and pharmacological regulation of the endocannabinoid CB1 receptor in Duchenne muscular dystrophy
Authors: Iannotti Fabio A. , Pagano Ester , Guardiola Ombretta , Adinolfi Simone , Saccone Valentina , Consalvi Silvia , Piscitelli Fabiana , Gazzerro Elisabetta , Busetto Giuseppe , Carrella Diego , Capasso Raffaele , Puri Pier Lorenzo , Minchiotti Gabriella , Di Marzo Vincenzo
Published: September 2018

The endocannabinoid system refers to a widespread signaling system and its alteration is implicated in a growing number of human diseases. However, the potential role of endocannabinoids in skeletal muscle disorders remains unknown. Here we report the role of the endocannabinoid CB1 receptors in Duchenne’s muscular dystrophy. In murine and human models, CB1 transcripts show the highest degree of expression at disease onset, and then decline overtime. Similar changes are observed for PAX7, a key regulator of muscle stem cells. Bioinformatics and biochemical analysis reveal that PAX7 binds and upregulates the CB1 gene in dystrophic more than in healthy muscles. Rimonabant, an antagonist of CB1, promotes human satellite cell differentiation in vitro, increases the number of regenerated myofibers, and prevents locomotor impairment in dystrophic mice. In conclusion, our study uncovers a PAX7–CB1 cross talk potentially exacerbating DMD and highlights the role of CB1 receptors as target for potential therapies.
Nuclear lamina dysfunction triggers a germline stem cell checkpoint
Authors: Barton Lacy J. , Duan Tingting , Ke Wenfan , Luttinger Amy , Lovander Kaylee E. , Soshnev Alexey A. , Geyer Pamela K.
Published: September 2018

LEM domain (LEM-D) proteins are conserved components of the nuclear lamina (NL) that contribute to stem cell maintenance through poorly understood mechanisms. The Drosophila emerin homolog Otefin (Ote) is required for maintenance of germline stem cells (GSCs) and gametogenesis. Here, we show that ote mutants carry germ cell-specific changes in nuclear architecture that are linked to GSC loss. Strikingly, we found that both GSC death and gametogenesis are rescued by inactivation of the DNA damage response (DDR) kinases, ATR and Chk2. Whereas the germline checkpoint draws from components of the DDR pathway, genetic and cytological features of the GSC checkpoint differ from the canonical pathway. Instead, structural deformation of the NL correlates with checkpoint activation. Despite remarkably normal oogenesis, rescued oocytes do not support embryogenesis. Taken together, these data suggest that NL dysfunction caused by Otefin loss triggers a GSC-specific checkpoint that contributes to maintenance of gamete quality.
Structural insights into modulation and selectivity of transsynaptic neurexin–LRRTM interaction
Authors: Yamagata Atsushi , Goto-Ito Sakurako , Sato Yusuke , Shiroshima Tomoko , Maeda Asami , Watanabe Masahiko , Saitoh Takashi , Maenaka Katsumi , Terada Tohru , Yoshida Tomoyuki , Uemura Takeshi , Fukai Shuya
Published: September 2018

Leucine-rich repeat transmembrane neuronal proteins (LRRTMs) function as postsynaptic organizers that induce excitatory synapses. Neurexins (Nrxns) and heparan sulfate proteoglycans have been identified as presynaptic ligands for LRRTMs. Specifically, LRRTM1 and LRRTM2 bind to the Nrxn splice variant lacking an insert at the splice site 4 (S4). Here, we report the crystal structure of the Nrxn1β–LRRTM2 complex at 3.4 Å resolution. The Nrxn1β–LRRTM2 interface involves Ca2+-mediated interactions and overlaps with the Nrxn–neuroligin interface. Together with structure-based mutational analyses at the molecular and cellular levels, the present structural analysis unveils the mechanism of selective binding between Nrxn and LRRTM1/2 and its modulation by the S4 insertion of Nrxn.
A Mendelian randomization study of the effects of blood lipids on breast cancer risk
Authors: Nowak Christoph , Ärnlöv Johan
Published: September 2018

Observational studies have reported inconsistent associations between circulating lipids and breast cancer risk. Using results from >400,000 participants in two-sample Mendelian randomization, we show that genetically raised LDL-cholesterol is associated with higher risk of breast cancer (odds ratio, OR, per standard deviation, 1.09, 95% confidence interval, 1.02–1.18, P = 0.020) and estrogen receptor (ER)-positive breast cancer (OR 1.14 [1.05–1.24] P = 0.004). Genetically raised HDL-cholesterol is associated with higher risk of ER-positive breast cancer (OR 1.13 [1.01–1.26] P = 0.037). HDL-cholesterol-raising variants in the gene encoding the target of CETP inhibitors are associated with higher risk of breast cancer (OR 1.07 [1.03–1.11] P = 0.001) and ER-positive breast cancer (OR 1.08 [1.03–1.13] P = 0.001). LDL-cholesterol-lowering variants mimicking PCSK9 inhibitors are associated (P = 0.014) with lower breast cancer risk. We find no effects related to the statin and ezetimibe target genes. The possible risk-promoting effects of raised LDL-cholesterol and CETP-mediated raised HDL-cholesterol have implications for breast cancer prevention and clinical trials.
Endurance of quantum coherence due to particle indistinguishability in noisy quantum networks
Authors: Perez-Leija Armando , Guzmán-Silva Diego , León-Montiel Roberto de J. , Gräfe Markus , Heinrich Matthias , Moya-Cessa Hector , Busch Kurt , Szameit Alexander
Published: September 2018

Quantum coherence, the physical property underlying fundamental phenomena such as multi-particle interference and entanglement, has emerged as a valuable resource upon which modern technologies are founded. In general, the most prominent adversary of quantum coherence is noise arising from the interaction of the associated dynamical system with its environment. Under certain conditions, however, the existence of noise may drive quantum and classical systems to endure intriguing nontrivial effects. In this vein, here we demonstrate, both theoretically and experimentally, that when two indistinguishable non-interacting particles co-propagate through quantum networks affected by non-dissipative noise, the system always evolves into a steady state in which coherences accounting for particle indistinguishabilty perpetually prevail. Furthermore, we show that the same steady state with surviving quantum coherences is reached even when the initial state exhibits classical correlations.
Serum deprivation/starvation leads to reactivation of HIV-1 in latently infected monocytes via activating ERK/JNK pathway
Authors: Raja Rameez , Lata Sneh , Trivedi Shubhendu , Banerjea Akhil C.
Published: September 2018

Despite the high success rate, antiretroviral therapy does not cure the disease completely due to presence of latent viral reservoirs. Although several studies have addressed this issue earlier, the role of serum starvation/deprivation in HIV-1 latency has not been studied. So, we investigated the role of serum starvation in regulating HIV-1 latency. The impact of serum starvation on HIV-1 latency was assessed in latently infected monocytes U1 and T-cells J1.1. Serum starvation breaks HIV-1 latency in U1 cells. Under similar conditions, J1.1 cells failed to show reactivation of virus. We investigated the involvement of cell death pathway and autophagy during the serum starvation in viral reactivation. Inhibition of these pathways did not affect viral reactivation. Furthermore, other crucial factors like NF-κB, SP1 and AKT did not play any role in regulating viral latency. Here, we report that serum deprivation up-regulates ERK/JNK pathway. This leads to phosphorylation of c-Jun which plays an important role in viral reactivation. Treatment of cells with U0126, an ERK kinase inhibitor, potently inhibited viral replication. In summary, we show that serum starvation leads to reactivation of HIV-1 in latently infected monocytes through the ERK/JNK pathway.
Author Correction: Abnormal contractility in human heart myofibrils from patients with dilated cardiomyopathy due to mutations in TTN and contractile protein genes
Authors: Vikhorev Petr G. , Smoktunowicz Natalia , Munster Alex B. , Copeland O’ Neal , Kostin Sawa , Montgiraud Cecile , Messer Andrew E. , Toliat Mohammad R. , Li Amy , dos Remedios Cristobal G. , Lal Sean , Blair Cheavar A. , Campbell Kenneth S. , Guglin Maya , Richter Manfred , Knöll Ralph , Marston Steven B.
Published: September 2018

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.
An emotional modulation model as signature for the identification of children developmental disorders
Authors: Mencattini Arianna , Mosciano Francesco , Comes Maria Colomba , Di Gregorio Tania , Raguso Grazia , Daprati Elena , Ringeval Fabien , Schuller Bjorn , Di Natale Corrado , Martinelli Eugenio
Published: September 2018

In recent years, applications like Apple’s Siri or Microsoft’s Cortana have created the illusion that one can actually “chat” with a machine. However, a perfectly natural human-machine interaction is far from real as none of these tools can empathize. This issue has raised an increasing interest in speech emotion recognition systems, as the possibility to detect the emotional state of the speaker. This possibility seems relevant to a broad number of domains, ranging from man-machine interfaces to those of diagnostics. With this in mind, in the present work, we explored the possibility of applying a precision approach to the development of a statistical learning algorithm aimed at classifying samples of speech produced by children with developmental disorders(DD) and typically developing(TD) children. Under the assumption that acoustic features of vocal production could not be efficiently used as a direct marker of DD, we propose to apply the Emotional Modulation function(EMF) concept, rather than running analyses on acoustic features per se to identify the different classes. The novel paradigm was applied to the French Child Pathological & Emotional Speech Database obtaining a final accuracy of 0.79, with maximum performance reached in recognizing language impairment (0.92) and autism disorder (0.82).
Improved Global Surface Temperature Simulation using Stratospheric Ozone Forcing with More Accurate Variability
Authors: Xie Fei , Li Jianping , Sun Cheng , Ding Ruiqiang , Xing Nan , Yang Yun , Zhou Xin , Ma Xuan
Published: September 2018

Increasingly, studies have pointed out that variations of stratospheric ozone significantly influence climate change in the Northern and Southern hemispheres. This leads us to consider whether making the variations of stratospheric ozone in a climate model closer to real ozone changes would improve the simulation of global climate change. It is found that replacing the original specified stratospheric ozone forcing with more accurate stratospheric ozone variations improves the simulated variations of surface temperature in a climate model. The improved stratospheric ozone variations in the Northern Hemisphere lead to better simulation of variations in Northern Hemisphere circulation. As a result, the simulated variabilities of surface temperature in the middle of the Eurasian continent and in lower latitudes are improved. In the Southern Hemisphere, improvements in surface temperature variations that result from improved stratospheric ozone variations influence the simulation of westerly winds. The simulations also suggest that the decreasing trend of stratospheric ozone may have enhanced the warming trend at high latitudes in the second half of the 20th century. Our results not only reinforce the importance of accurately simulating the stratospheric ozone but also imply the need for including fully coupled stratospheric dynamical–radiative–chemical processes in climate models to predict future climate changes.
Climate change-driven range losses among bumblebee species are poised to accelerate
Authors: Sirois-Delisle Catherine , Kerr Jeremy T.
Published: September 2018

Climate change has shaped bee distributions over the past century. Here, we conducted the first species-specific assessment of future climate change impacts on North American bumblebee distributions, using the most recent global change scenarios developed in the Fifth Assessment Report of the Intergovernmental Panel on Climate Change (IPCC). We assessed potential shifts in bumblebee species distributions with models generated using Maxent. We tested different assumptions about bumblebee species’ dispersal capacities, drawing on observed patterns of range shifts to date, dispersal rates observed for bumblebee queens, and, lastly, assuming unlimited dispersal. Models show significant contractions of current ranges even under scenarios in which dispersal rates were high. Results suggest that dispersal rates may not suffice for bumblebees to track climate change as rapidly as required under any IPCC scenario for future climate change. Areas where species losses are projected overlap for many species and climate scenarios, and are concentrated in eastern parts of the continent. Models also show overlap for range expansions across many species, suggesting the presence of “hotspots” where management activities could benefit many species, across all climate scenarios. Broad-scale strategies are likely to be necessary to improve bumblebee conservation prospects under climate change.
Aphid infestation in the phyllosphere affects primary metabolic profiles in the arbuscular mycorrhizal hyphosphere
Authors: Cabral Carmina , Wollenweber Bernd , António Carla , Rodrigues Ana Margarida , Ravnskov Sabine
Published: September 2018

While effects of (a)biotic stress events in the phyllosphere have been studied intensively, possible influences of stress on the arbuscular mycorrhizal hyphosphere has scarcely been investigated. We hypothesised that stress challenge in the phyllosphere could alter primary metabolite profiles of the hyphosphere - the mycelial network connecting plants. Donor plants, connected to receiver plants by mycelial networks, were aphid-challenged during 84 h. Primary metabolite profiles in the hyphosphere were investigated. Gene-expression of plant defence gene PR1 was measured in one of the receiver plants during the challenge. Hexose levels in the hyphosphere increased when donor plants were aphid-challenged. This change in metabolic profile was influenced by leaf sampling from receiver plant. PR1 expression increased in donor plants 48 h after challenge, and consequently 60 h after, in receiver plants. We conclude that aphid infestation of donor plants modified primary carbon metabolism in the hyphosphere. Plant defence response in receiver plants, occurred 12 h after detection of response in the aphid-challenged donor plants. While this work is the first to reveal primary metabolic profiles of the AM hyphosphere, more work is needed to elucidate the possible role of transient changes of hexose metabolism in stress response and signalling processes in the hyphosphere of connected plants.
Preparation and characterization of thermo-sensitive gel with phenolated alkali lignin
Authors: Jiang Pan , Sheng Xueru , Yu Sheng , Li Haiming , Lu Jie , Zhou Jinghui , Wang Haisong
Published: September 2018

Thermo-sensitive gel exhibits great potential industrial application. It has been widely used in tissue repair, drug release and water purification for its property of phase transition in response to external stimuli, reusability and biocompatibility. In this study, a novel lignin-based thermo-sensitive gel was synthesized with alkali lignin by two steps. Firstly, phenolated lignin (PPAL) was synthesized with purified alkali lignin (PAL) catalyzed by sulfuric acid. Subsequently, thermo-sensitive gel was achieved by thermal polymerization of phenolated alkali lignin and N-isopropylacrylamide (NIPAAm). Furthermore, the prepared hydrogels were characterized in terms of thermal behavior, interior morphology and their swelling behavior. Compared with PAL-based gel, the obtained PPAL-based gel exhibits a higher crosslinking density and lower critical solution temperature (LCST) due to the increase of reaction site and smaller space volume of the hydrophobic side groups grafted on NIPAAm. TGA data revealed that thermal stability of gel was enhanced (50% weight loss at ~380 °C) by using lignin as precursor. SEM images showed that a more regular interior morphology, better compressive strength was also found (PPAL0.05, 11.15 KPa). Furthermore, the swelling ratio of PPAL-based gel was lower than that of PAL-based gel due to its more complex structure.
Baicalin, the major component of traditional Chinese medicine Scutellaria baicalensis induces colon cancer cell apoptosis through inhibition of oncomiRNAs
Authors: Tao Yili , Zhan Shoubin , Wang Yanbo , Zhou Geyu , Liang Hongwei , Chen Xi , Shen Hong
Published: September 2018

Colorectal cancer (CRC) is among the most frequently occurring cancers worldwide. Baicalin is isolated from the roots of Scutellaria baicalensis and is its dominant flavonoid. Anticancer activity of baicalin has been evaluated in different types of cancers, especially in CRC. However, the molecular mechanisms underlying the contribution of baicalin to the treatment of CRC are still unknown. Here, we confirmed that baicalin can effectively induce and enhance apoptosis in HT-29 cells in a dose-dependent manner and suppress tumour growth in xenografted nude mice. We further performed a miRNA microarray analysis of baicalin-treated and untreated HT-29 cells. The results showed that a large number of oncomiRs, including miR-10a, miR-23a, miR-30c, miR-31, miR-151a and miR-205, were significantly suppressed in baicalin-treated HT-29 cells. Furthermore, our in vitro and in vivo studies showed that baicalin suppressed oncomiRs by reducing the expression of c-Myc. Taken together, our study shows a novel mechanism for anti-cancer action of baicalin, that it induces apoptosis in colon cancer cells and suppresses tumour growth by reducing the expression of c-Myc and oncomiRs.
Parallelized, real-time, metabolic-rate measurements from individual Drosophila
Authors: Fiorino Anthony , Thompson Dakotah , Yadlapalli Swathi , Jiang Chang , Shafer Orie. T. , Reddy Pramod , Meyhofer Edgar
Published: September 2018

Significant recent evidence suggests that metabolism is intricately linked to the regulation and dysfunction of complex cellular and physiological responses ranging from altered metabolic programs in cancers and aging to circadian rhythms and molecular clocks. While the metabolic pathways and their fundamental control mechanisms are well established, the precise cellular mechanisms underpinning, for example, enzymatic pathway control, substrate preferences or metabolic rates, remain far less certain. Comprehensive, continuous metabolic studies on model organisms, such as the fruit fly Drosophila melanogaster, may provide a critical tool for deciphering these complex physiological responses. Here, we describe the development of a high-resolution calorimeter, which combines sensitive thermometry with optical imaging to concurrently perform measurements of the metabolic rate of ten individual flies, in real-time, with ~100 nW resolution. Using this calorimeter we have measured the mass-specific metabolic rates of flies of different genotypes, ages, and flies fed with different diets. This powerful new approach enables systematic studies of the metabolic regulation related to cellular and physiological function and disease mechanisms.
Bioprocessing strategies to enhance the challenging isolation of neuro-regenerative cells from olfactory mucosa
Authors: Georgiou Melanie , Reis Joana Neves dos , Wood Rachael , Esteban Patricia Perez , Roberton Victoria , Mason Chris , Li Daqing , Li Ying , Choi David , Wall Ivan
Published: September 2018

Olfactory ensheathing cells (OECs) are a promising potential cell therapy to aid regeneration. However, there are significant challenges in isolating and characterizing them. In the current study, we have explored methods to enhance the recovery of cells expressing OEC marker p75NTR from rat mucosa. With the addition of a 24-hour differential adhesion step, the expression of p75NTR was significantly increased to 73 ± 5% and 46 ± 18% on PDL and laminin matrices respectively. Additionally, the introduction of neurotrophic factor NT-3 and the decrease in serum concentration to 2% FBS resulted in enrichment of OECs, with p75NTR at nearly 100% (100 ± 0% and 98 ± 2% on PDL and laminin respectively), and candidate fibroblast marker Thy1.1 decreased to zero. Culturing OECs at physiologically relevant oxygen tension (2–8%) had a negative impact on p75NTR expression and overall cell survival. Regarding cell potency, co-culture of OECs with NG108-15 neurons resulted in more neuronal growth and potential migration at atmospheric oxygen. Moreover, OECs behaved similarly to a Schwann cell line positive control. In conclusion, this work identified key bioprocessing fundamentals that will underpin future development of OEC-based cell therapies for potential use in spinal cord injury repair. However, there is still much work to do to create optimized isolation methods.
The critical role of dietary foliage in maintaining the gut microbiome and metabolome of folivorous sifakas
Authors: Greene Lydia K. , McKenney Erin A. , O’Connell Thomas M. , Drea Christine M.
Published: September 2018

The gut microbiome (GMB) of folivores metabolizes dietary fiber into nutrients, including short-chain fatty acids (SCFAs); however, experiments probing the consequences of foliage quality on host GMBs are lacking. We therefore examined GMB structure and function via amplicon sequencing and Nuclear Magnetic Resonance spectroscopy in 31 captive sifakas (Propithecus coquereli) during dietary manipulations associated with husbandry. Supplementing standard diets with diverse foliage blends, versus with a single plant species, promoted more diverse GMBs, enriched for taxa implicated in plant-fiber metabolism, but depleted in taxa implicated in starch metabolism and bile tolerance. The consumption of diverse blends was associated with greater concentrations of colonic SCFAs. Abundant foliage, via forest access, promoted compositionally distinct and more stable GMBs, but reduced concentrations of SCFAs, possibly reflecting selection of high-quality leaves. In 11 subjects denied forest access, we examined the temporal pace of microbial shifts when supplemental foliage was abruptly switched between diverse blends and single species. The sifaka GMB responded within days, with community diversity and composition closely tracking foliage diversity. By providing experimental evidence that the folivore GMB is sensitive to minor changes in dietary foliage, we reveal the fragility of specialist GMBs, with implications for managing the wellbeing of endangered wildlife.
Shear Wave Elasticity Measurements of Three-Dimensional Cancer Cell Cultures Using Laser Speckle Contrast Imaging
Authors: Chao Pei-Yu , Liu Wei-Wen , You Shih-Feng , Li Pai-Chi
Published: September 2018

Shear wave elastography (SWE) has been widely adopted for clinical in vivo imaging of tissue elasticity for disease diagnosis, and this modality can be a valuable tool for in vitro mechanobiology studies but its full potential has yet to be explored. Here we present a laser speckle contrast SWE system for noncontact monitoring the spatiotemporal changes of the extracellular matrix (ECM) stiffness in three-dimensional cancer cell culture system while providing submillimeter spatial resolution and temporal resolution of 10 s. The shear modulus measured was found to be strongly correlated with the ECM fiber density in two types of cell culture system (r = 0.832 with P < 0.001, and r = 0.642 with P = 0.024 for cell culture systems containing 4 mg/ml Matrigel with 1 mg/ml and 2 mg/ml collagen type I hydrogel, respectively). Cell migration along the stiffness gradient in the cell culture system and an association between cell proliferation and the local ECM stiffness was observed. As the elasticity measurement is performed without the need of exogenous probes, the proposed method can be used to study how the microenvironmental stiffness interacts with cancer cell behaviors without possible adverse effects of the exogenous particles, and could potentially be an effective screening tool when developing new treatment strategies.
Impact of deep ocean mixing on the climatic mean state in the Southern Ocean
Authors: Tatebe Hiroaki , Tanaka Yuki , Komuro Yoshiki , Hasumi Hiroyasu
Published: September 2018

The Southern Ocean is of great importance for the global stratification and biological carbon storage because it is connected to the global ocean conveyor by which atmospheric information absorbed in the Southern Ocean is redistributed globally and buffered over centuries. Therefore, understanding what controls the Southern Ocean climate, the global ocean conveyor, and links between them is a key to quantifying uncertainties in future climate projections. Based on a set of climate model experiments, here we show that the tide-induced micro-scale mixing in the Pacific deep ocean has significant impacts on the wintertime Southern Ocean climate through basin-scale reorganization of ocean stratification and resultant response of the global ocean conveyor. Specifically, Pacific deep water, which is modified by the deep ocean mixing while travelling south, reinforces the subsurface stratification and suppresses deep convection in the Southern Ocean. Resultant increase of the Ross Sea sea-ice leads to decrease of incoming shortwave radiation and strengthening of the westerly and storms. Because the Southern Ocean could regulate the global warming progress through its role as heat and carbon sink, our study implies that better representation of deep ocean mixing in climate models contributes to reliability improvement in regional-to-global climate projections.
Circadian rhythmicity persists through the Polar night and midnight sun in Svalbard reindeer
Authors: Arnold Walter , Ruf Thomas , Loe Leif Egil , Irvine R. Justin , Ropstad Erik , Veiberg Vebjørn , Albon Steve D.
Published: September 2018

Studies of locomotor activity in Svalbard reindeer reported the temporary absence of diel rhythms under Arctic photic conditions. However, using Lomb-Scargle periodogram analyses with high statistical power we found diel or circadian rhythmicity throughout the entire year in measures of behaviour, temperature in the rumen and heart rate in free-living Svalbard reindeer. Significant diel rhythmicity was only lacking during some of the 15-day intervals analysed in the less frequently measured heart rate. During Polar Night these rhythms were free-running and attenuated. During continual daylight in summer, rhythms where entrained to 24 hours corresponding with the daily variation in the intensity of solar radiation, but attenuated when continuous daylight coincided with the period of growing forage. Diel rhythmicity was reduced during this short period of peak foraging activity, which coincided with peak heart rate and temperature in the rumen, most likely to facilitate fattening when food is abundant. For the rest of the year, heart rate and temperature showed the most pronounced and long-lasting suppression ever found in ungulates. The profound seasonal changes in foraging, metabolic activity, and power of diel and circadian rhythmicity of Svalbard reindeer can be viewed as adaptations to the extreme living conditions in the High Arctic.
Paraoxonase 1 knockout rats have impaired T cell development at the CD4/CD8 double-negative to double-positive transition stage
Authors: Bai Lin , Shi Guiying , Ma Yuanwu , Zhang Li , Guan Feifei , Zhang Xu , Xu Yanfeng , Chen Houzao , Zhang Lianfeng
Published: September 2018

Paraoxonase 1 (PON1) is a high-density lipoprotein-associated enzyme that performs multiple physiological activities. Previous studies suggest that PON1 plays an anti-inflammatory role in the cardiovascular system, although its roles in hematopoiesis and adaptive immunity have not been clarified. To investigate the impact of PON1 on the immune system, we generated PON1-knockout (PON1−/−) rats using the CRISPR/Cas9 system. The thymus was smaller in PON1−/− rats than that in wild-type (PON1+/+) rats. Furthermore, analysis of thymocyte development revealed diminished total T cell numbers and a decrease in CD4+, CD8+ and double-positive T cells in peripheral blood and thymus from PON1−/− rats. This may be due to a block in the transition of T cells from the double-negative to the double-positive stage. We also showed that the activation of p38 MAPK phosphorylation contributed to the increased apoptosis and defective T cell development in PON−/− rats. Therefore, our results indicate that PON1 functions as a novel regulator of T cell development.
A single reporter mouse line for Vika, Flp, Dre, and Cre-recombination
Authors: Karimova Madina , Baker Oliver , Camgoz Aylin , Naumann Ronald , Buchholz Frank , Anastassiadis Konstantinos
Published: September 2018

Site-specific recombinases (SSR) are utilized as important genome engineering tools to precisely modify the genome of mice and other model organisms. Reporter mice that mark cells that at any given time had expressed the enzyme are frequently used for lineage tracing and to characterize newly generated mice expressing a recombinase from a chosen promoter. With increasing sophistication of genome alteration strategies, the demand for novel SSR systems that efficiently and specifically recombine their targets is rising and several SSR-systems are now used in combination to address complex biological questions in vivo. Generation of reporter mice for each one of these recombinases is cumbersome and increases the number of mouse lines that need to be maintained in animal facilities. Here we present a multi-reporter mouse line for loci-of-recombination (X) (MuX) that streamlines the characterization of mice expressing prominent recombinases. MuX mice constitutively express nuclear green fluorescent protein after recombination by either Cre, Flp, Dre or Vika recombinase, rationalizing the number of animal lines that need to be maintained. We also pioneer the use of the Vika/vox system in mice, illustrating its high efficacy and specificity, thereby facilitating future designs of sophisticated recombinase-based in vivo genome engineering strategies.
Correlated Light-Serial Scanning Electron Microscopy (CoLSSEM) for ultrastructural visualization of single neurons in vivo
Authors: Hirabayashi Yusuke , Tapia Juan Carlos , Polleux Franck
Published: September 2018

A challenging aspect of neuroscience revolves around mapping the synaptic connections within neural circuits (connectomics) over scales spanning several orders of magnitude (nanometers to meters). Despite significant improvements in serial section electron microscopy (SSEM) technologies, several major roadblocks have impaired its general applicability to mammalian neural circuits. In the present study, we introduce a new approach that circumvents some of these roadblocks by adapting a genetically-encoded ascorbate peroxidase (APEX2) as a fusion protein to a membrane-targeted fluorescent reporter (CAAX-Venus), and introduce it in single pyramidal neurons in vivo using extremely sparse in utero cortical electroporation. This approach allows us to perform Correlated Light-SSEM (CoLSSEM), a variant of Correlated Light-EM (CLEM), on individual neurons, reconstructing their dendritic and axonal arborization in a targeted way via combination of high-resolution confocal microscopy, and subsequent imaging of its ultrastructural features and synaptic connections with ATUM-SEM (automated tape-collecting ultramicrotome - scanning electron microscopy) technology. Our method significantly will improve the feasibility of large-scale reconstructions of neurons within a circuit, and permits the description of some ultrastructural features of identified neurons with their functional and/or structural connectivity, one of the main goal of connectomics.
Identification of Peptide Mimotope Ligands for Natalizumab
Authors: Ruff Laura E. , Pfeilsticker Jessica A. , Johnsen Nicholas E. , Nocchi Sarah , Messmer Bradley T.
Published: September 2018

Mimotope peptides selected from combinatorial peptide libraries can be used as capture reagents for immunoassay detection of therapeutic monoclonal antibodies (mAbs). We report the use of phage display libraries to identify peptide ligands (VeritopesTM) that bind natalizumab, a therapeutic mAb indicated for use in multiple sclerosis. PKNPSKF is identified as a novel natalizumab-binding motif, and peptides containing this motif demonstrated utility as capture reagents in enzyme-linked immunosorbent assays (ELISAs). A peptide containing the identified motif was shown to be competitive with the natural ligand (α4-integrin) and a neutralizing anti-idiotype antibody for natalizumab binding, indicating that VeritopesTM act as surrogate ligands that bind the antigen binding site of natalizumab. Affinity maturation further confirmed the motif sequence and yielded peptides with greater apparent affinity by ELISA. VeritopesTM are promising assay reagents for therapeutic drug level monitoring.
Analysis of the association of EPHB6, EFNB1 and EFNB3 variants with hypertension risks in males with hypogonadism
Authors: Wu Tao , Zhang Bi-Qi , Raelson John , Yao Yu-Mei , Wu Huan-Dong , Xu Zao-Xian , Marois-blanchet Francois-christophe , Tahir Muhammad Ramzan , Wang Yujia , Bradley W. Edward , Luo Hongyu , Wu Jiangping , Sheng Jian-Zhong , Hu Shen-Jiang
Published: September 2018

Several members of the EPH kinase family and their ligands are involved in blood pressure regulation, and such regulation is often sex- or sex hormone-dependent, based on animal and human genetic studies. EPHB6 gene knockout (KO) in mice leads to hypertension in castrated males but not in un-manipulated KO males or females. To assess whether this finding in mice is relevant to human hypertension, we conducted a human genetic study for the association of EPHB6 and its two ligands, EFNB1 and EFNB3, with hypertension in hypogonadic patients. Seven hundred and fifty hypertensive and 750 normotensive Han Chinese patients, all of whom were hypogonadic, were genotyped for single nucleotide polymorphisms (SNPs) within the regions of the genes, plus an additional 50 kb 5′ of the genes for EPHB6, EFNB1 and EFNB3. An imputed insertion/deletion polymorphism, rs35530071, was found to be associated with hypertension at p-values below the Bonferroni-corrected significance level of 0.0024. This marker is located 5′ upstream of the EFNB3 gene start site. Previous animal studies showed that while male EFNB3 gene knockout mice were normotensive, castration of these mice resulted in hypertension, corroborating the results of the human genetic study. Considering the significant associations of EFNB3 SNPs with hypertension in hypogonadic males and supporting evidence from castrated EFNB3 KO mice, we conclude that loss-of-function variants of molecules in the EPHB6 signaling pathway in the presence of testosterone are protective against hypertension in humans.
Genome-wide burden and association analyses implicate copy number variations in asthma risk among children and young adults from Latin America
Authors: Oliveira Pablo , Costa Gustavo N. O. , Damasceno Andresa K. A. , Hartwig Fernando P. , Barbosa George C. G. , Figueiredo Camila A. , Ribeiro-Silva Rita de C. , Pereira Alexandre , Lima-Costa M. Fernanda , Kehdy Fernanda S. , Tarazona-Santos Eduardo , Horta Bernardo L. , Rodrigues Laura C. , Fiaccone Rosemeire L. , Barreto Maurício L.
Published: September 2018

The genetic architecture of asthma was relatively well explored. However, some work remains in the field to improve our understanding on asthma genetics, especially in non-Caucasian populations and with regards to commonly neglected genetic variants, such as Copy Number Variations (CNVs). In the present study, we investigated the contribution of CNVs on asthma risk among Latin Americans. CNVs were inferred from SNP genotyping data. Genome wide burden and association analyses were conducted to evaluate the impact of CNVs on asthma outcome. We found no significant difference in the numbers of CNVs between asthmatics and non-asthmatics. Nevertheless, we found that CNVs are larger in patients then in healthy controls and that CNVs from cases intersect significantly more genes and regulatory elements. We also found that a deletion at 6p22.1 is associated with asthma symptoms in children from Salvador (Brazil) and in young adults from Pelotas (Brazil). To support our results, we conducted an in silico functional analysis and found that this deletion spans several regulatory elements, including two promoter elements active in lung cells. In conclusion, we found robust evidence that CNVs could contribute for asthma susceptibility. These results uncover a new perspective on the influence of genetic factors modulating asthma risk.
MoB2 Driven Metallic Behavior and Interfacial Charge Transport Mechanism in MoS2/MoB2 Heterostructure: A First-Principles Study
Authors: Bano Amreen , Pandey Devendra K. , Modi Anchit , Gaur N. K.
Published: September 2018

We have performed the density functional theory calculations on heterostructure (HS) of MoS2 and MoB2 monolayers. The aim of this study is to assess the influence of MoB2 on electron transport of adjacent MoS2 layer. In present investigation we predict that the electronic properties of MoS2 monolayer is influenced by 4d-states of Mo in MoB2 monolayer. Whereas, the B atoms of MoB2 and S atoms of MoS2 exhibit overlapping of intermediate atomic orbitals thereby collectively construct the interfacial electronic structure observed to be metallic in nature. From charge density calculations, we have also determine that the charge transfer is taking place at the interface via B-2p and S-3p states. The bonds at the interface are found to be metallic which is also confirmed by adsorption analysis. Thermoelectric performance of this HS is found be in good agreement with available literature. Low Seebeck coefficient and high electrical conductivity further confirms the existence of metallic state of the HS.