bluuebunny/crossref_metadata_2025 huggingface dataset:
The origin of heterogeneous nanoparticle uptake by cells
Authors: Rees Paul , Wills John W. , Brown M. Rowan , Barnes Claire M. , Summers Huw D.
Published: May 2019

Understanding nanoparticle uptake by biological cells is fundamentally important to wide-ranging fields from nanotoxicology to drug delivery. It is now accepted that the arrival of nanoparticles at the cell is an extremely complicated process, shaped by many factors including unique nanoparticle physico-chemical characteristics, protein-particle interactions and subsequent agglomeration, diffusion and sedimentation. Sequentially, the nanoparticle internalisation process itself is also complex, and controlled by multiple aspects of a cell’s state. Despite this multitude of factors, here we demonstrate that the statistical distribution of the nanoparticle dose per endosome is independent of the initial administered dose and exposure duration. Rather, it is the number of nanoparticle containing endosomes that are dependent on these initial dosing conditions. These observations explain the heterogeneity of nanoparticle delivery at the cellular level and allow the derivation of simple, yet powerful probabilistic distributions that accurately predict the nanoparticle dose delivered to individual cells across a population.
Tuning the permselectivity of polymeric desalination membranes via control of polymer crystallite size
Authors: Lu Xinglin , Feng Xunda , Yang Yi , Jiang Jin , Cheng Wei , Liu Caihong , Gopinadhan Manesh , Osuji Chinedum O. , Ma Jun , Elimelech Menachem
Published: May 2019

Membrane desalination is a leading technology for treating saline waters to augment fresh water supply. The need for high-performance desalination membranes, particularly with high water/salt selectivity, has stimulated research into the fundamental structure-property-performance relationship of state-of-the-art membranes. In this study, we utilize a facile method for tuning properties of a polymeric desalination membrane to shed light on water and salt transport mechanisms of such membranes. A desalination membrane made of cellulose triacetate is treated in a plasticizer solution, followed by water rinsing. The modified membranes exhibit reduced salt flux without compromising water flux, indicating enhanced water/salt selectivity. An inspection of material characteristics using a model film system reveals a plasticizing-extracting process in changing the polymeric structure, which leads to the reduction of crystallite size in the polymer matrix, consequently affecting the transport properties of the membranes. Our findings highlight the potential of the plasticizing-extracting process in fabricating membranes with desired desalination performance.
FANCM limits ALT activity by restricting telomeric replication stress induced by deregulated BLM and R-loops
Authors: Silva Bruno , Pentz Richard , Figueira Ana Margarida , Arora Rajika , Lee Yong Woo , Hodson Charlotte , Wischnewski Harry , Deans Andrew J. , Azzalin Claus M.
Published: May 2019

Telomerase negative immortal cancer cells elongate telomeres through the Alternative Lengthening of Telomeres (ALT) pathway. While sustained telomeric replicative stress is required to maintain ALT, it might also lead to cell death when excessive. Here, we show that the ATPase/translocase activity of FANCM keeps telomeric replicative stress in check specifically in ALT cells. When FANCM is depleted in ALT cells, telomeres become dysfunctional, and cells stop proliferating and die. FANCM depletion also increases ALT-associated marks and de novo synthesis of telomeric DNA. Depletion of the BLM helicase reduces the telomeric replication stress and cell proliferation defects induced by FANCM inactivation. Finally, FANCM unwinds telomeric R-loops in vitro and suppresses their accumulation in cells. Overexpression of RNaseH1 completely abolishes the replication stress remaining in cells codepleted for FANCM and BLM. Thus, FANCM allows controlled ALT activity and ALT cell proliferation by limiting the toxicity of uncontrolled BLM and telomeric R-loops.
The FANCM-BLM-TOP3A-RMI complex suppresses alternative lengthening of telomeres (ALT)
Authors: Lu Robert , O’Rourke Julienne J. , Sobinoff Alexander P. , Allen Joshua A. M. , Nelson Christopher B. , Tomlinson Christopher G. , Lee Michael , Reddel Roger R. , Deans Andrew J. , Pickett Hilda A.
Published: May 2019

The collapse of stalled replication forks is a major driver of genomic instability. Several committed mechanisms exist to resolve replication stress. These pathways are particularly pertinent at telomeres. Cancer cells that use Alternative Lengthening of Telomeres (ALT) display heightened levels of telomere-specific replication stress, and co-opt stalled replication forks as substrates for break-induced telomere synthesis. FANCM is a DNA translocase that can form independent functional interactions with the BLM-TOP3A-RMI (BTR) complex and the Fanconi anemia (FA) core complex. Here, we demonstrate that FANCM depletion provokes ALT activity, evident by increased break-induced telomere synthesis, and the induction of ALT biomarkers. FANCM-mediated attenuation of ALT requires its inherent DNA translocase activity and interaction with the BTR complex, but does not require the FA core complex, indicative of FANCM functioning to restrain excessive ALT activity by ameliorating replication stress at telomeres. Synthetic inhibition of FANCM-BTR complex formation is selectively toxic to ALT cancer cells.
Blimp1+ cells generate functional mouse sebaceous gland organoids in vitro
Authors: Feldman Alona , Mukha Dzmitry , Maor Itzhak I. , Sedov Egor , Koren Elle , Yosefzon Yahav , Shlomi Tomer , Fuchs Yaron
Published: May 2019

Most studies on the skin focus primarily on the hair follicle and interfollicular epidermis, whereas little is known regarding the homeostasis of the sebaceous gland (SG). The SG has been proposed to be replenished by different pools of hair follicle stem cells and cells that resides in the SG base, marked by Blimp1. Here, we demonstrate that single Blimp1+ cells isolated from mice have the potential to generate SG organoids in vitro. Mimicking SG homeostasis, the outer layer of these organoids is composed of proliferating cells that migrate inward, undergo terminal differentiation and generating lipid-filled sebocytes. Performing confocal microscopy and mass-spectrometry, we report that these organoids exhibit known markers and a lipidomic profile similar to SGs in vivo. Furthermore, we identify a role for c-Myc in sebocyte proliferation and differentiation, and determine that SG organoids can serve as a platform for studying initial stages of acne vulgaris, making this a useful platform to identify potential therapeutic targets.
SORLA regulates endosomal trafficking and oncogenic fitness of HER2
Authors: Pietilä Mika , Sahgal Pranshu , Peuhu Emilia , Jäntti Niklas Z. , Paatero Ilkka , Närvä Elisa , Al-Akhrass Hussein , Lilja Johanna , Georgiadou Maria , Andersen Olav M. , Padzik Artur , Sihto Harri , Joensuu Heikki , Blomqvist Matias , Saarinen Irena , Boström Peter J. , Taimen Pekka , Ivaska Johanna
Published: May 2019

The human epidermal growth factor receptor 2 (HER2) is an oncogene targeted by several kinase inhibitors and therapeutic antibodies. While the endosomal trafficking of many other receptor tyrosine kinases is known to regulate their oncogenic signalling, the prevailing view on HER2 is that this receptor is predominantly retained on the cell surface. Here, we find that sortilin-related receptor 1 (SORLA; SORL1) co-precipitates with HER2 in cancer cells and regulates HER2 subcellular distribution by promoting recycling of the endosomal receptor back to the plasma membrane. SORLA protein levels in cancer cell lines and bladder cancers correlates with HER2 levels. Depletion of SORLA triggers HER2 targeting to late endosomal/lysosomal compartments and impairs HER2-driven signalling and in vivo tumour growth. SORLA silencing also disrupts normal lysosome function and sensitizes anti-HER2 therapy sensitive and resistant cancer cells to lysosome-targeting cationic amphiphilic drugs. These findings reveal potentially important SORLA-dependent endosomal trafficking-linked vulnerabilities in HER2-driven cancers.
Structure of the SARS-CoV nsp12 polymerase bound to nsp7 and nsp8 co-factors
Authors: Kirchdoerfer Robert N. , Ward Andrew B.
Published: May 2019

Recent history is punctuated by the emergence of highly pathogenic coronaviruses such as SARS- and MERS-CoV into human circulation. Upon infecting host cells, coronaviruses assemble a multi-subunit RNA-synthesis complex of viral non-structural proteins (nsp) responsible for the replication and transcription of the viral genome. Here, we present the 3.1 Å resolution structure of the SARS-CoV nsp12 polymerase bound to its essential co-factors, nsp7 and nsp8, using single particle cryo-electron microscopy. nsp12 possesses an architecture common to all viral polymerases as well as a large N-terminal extension containing a kinase-like fold and is bound by two nsp8 co-factors. This structure illuminates the assembly of the coronavirus core RNA-synthesis machinery, provides key insights into nsp12 polymerase catalysis and fidelity and acts as a template for the design of novel antiviral therapeutics.
Synergistic and antagonistic impacts of suspended sediments and thermal stress on corals
Authors: Fisher Rebecca , Bessell-Browne Pia , Jones Ross
Published: May 2019

Understanding pressure pathways and their cumulative impacts is critical for developing effective environmental policy. For coral reefs, wide spread bleaching resulting from global warming is occurring concurrently with local pressures, such as increases in suspended sediments through coastal development. Here we examine the relative importance of suspended sediment pressure pathways for dredging impacts on corals and evidence for synergistic or antagonistic cumulative effects between suspended sediments and thermal stress. We show that low to moderate reductions in available light associated with dredging may lead to weak antagonistic (less than expected independently) cumulative effects. However, when sediment loads are high any reductions in mortality associated with reduced bleaching are outweighed by increased mortality associated with severe low light periods and high levels of sediment deposition and impacts become synergistic (greater than what would occur independently). The findings suggest efforts to assess global cumulative impacts need to consider how pressures interact to impact ecosystems, and that the cumulative outcome may vary across the range of realised pressure fields.
Infinitesimal sulfur fusion yields quasi-metallic bulk silicon for stable and fast energy storage
Authors: Ryu Jaegeon , Seo Ji Hui , Song Gyujin , Choi Keunsu , Hong Dongki , Wang Chongmin , Lee Hosik , Lee Jun Hee , Park Soojin
Published: May 2019

A fast-charging battery that supplies maximum energy is a key element for vehicle electrification. High-capacity silicon anodes offer a viable alternative to carbonaceous materials, but they are vulnerable to fracture due to large volumetric changes during charge–discharge cycles. The low ionic and electronic transport across the silicon particles limits the charging rate of batteries. Here, as a three-in-one solution for the above issues, we show that small amounts of sulfur doping (<1 at%) render quasi-metallic silicon microparticles by substitutional doping and increase lithium ion conductivity through the flexible and robust self-supporting channels as demonstrated by microscopy observation and theoretical calculations. Such unusual doping characters are enabled by the simultaneous bottom-up assembly of dopants and silicon at the seed level in molten salts medium. This sulfur-doped silicon anode shows highly stable battery cycling at a fast-charging rate with a high energy density beyond those of a commercial standard anode.
Endophilin-A2 dependent VEGFR2 endocytosis promotes sprouting angiogenesis
Authors: Genet Gael , Boyé Kevin , Mathivet Thomas , Ola Roxana , Zhang Feng , Dubrac Alexandre , Li Jinyu , Genet Nafiisha , Henrique Geraldo Luiz , Benedetti Lorena , Künzel Steffen , Pibouin-Fragner Laurence , Thomas Jean-Leon , Eichmann Anne
Published: May 2019

Endothelial cell migration, proliferation and survival are triggered by VEGF-A activation of VEGFR2. However, how these cell behaviors are regulated individually is still unknown. Here we identify Endophilin-A2 (ENDOA2), a BAR-domain protein that orchestrates CLATHRIN-independent internalization, as a critical mediator of endothelial cell migration and sprouting angiogenesis. We show thatEndoA2knockout mice exhibit postnatal angiogenesis defects and impaired front-rear polarization of sprouting tip cells. ENDOA2 deficiency reduces VEGFR2 internalization and inhibits downstream activation of the signaling effector PAK but not ERK, thereby affecting front-rear polarity and migration but not proliferation or survival. Mechanistically, VEGFR2 is directed towards ENDOA2-mediated endocytosis by the SLIT2-ROBO pathway via SLIT-ROBO-GAP1 bridging of ENDOA2 and ROBO1. Blocking ENDOA2-mediated endothelial cell migration attenuates pathological angiogenesis in oxygen-induced retinopathy models. This work identifies a specific endocytic pathway controlling a subset of VEGFR2 mediated responses that could be targeted to prevent excessive sprouting angiogenesis in pathological conditions.
Assessing spirituality: is there a beneficial role in the management of COPD?
Authors: Gergianaki Irini , Kampouraki Maria , Williams Siân , Tsiligianni Ioanna
Published: May 2019

Here,we report on the role of spirituality assessment in the management of chronic obstructive pulmonary disease (COPD). Although a positive effect of addressing spirituality in health care has been proved in a number of chronic diseases, its potential in COPD has received less attention. Although limited, available evidence suggests that spirituality may play an important role in improving quality of life of patients with COPD. The fruitful results in other long-term conditions may lay the foundation for further research on addressing spirituality in COPD. This should focus where the burden of COPD is greatest, including low-resource settings globally. Implementation research should include exploration of an acceptable consultation process to identify patients who would welcome spiritual discussions; how to integrate spiritual approaches into health care professional curricula so that they are aware of its importance and have the confidence to raise it with patients and how to integrate spiritual approaches into holistic COPD care.
Clinical courses and complications of young adults with Autosomal Recessive Polycystic Kidney Disease (ARPKD)
Authors: Burgmaier Kathrin , Kilian Samuel , Bammens Bert , Benzing Thomas , Billing Heiko , Büscher Anja , Galiano Matthias , Grundmann Franziska , Klaus Günter , Mekahli Djalila , Michel-Calemard Laurence , Milosevski-Lomic Gordana , Ranchin Bruno , Sauerstein Katja , Schaefer Susanne , Shroff Rukshana , Sterenborg Rosalie , Verbeeck Sarah , Weber Lutz T. , Wicher Dorota , Wühl Elke , Dötsch Jörg , Schaefer Franz , Liebau Max C.
Published: May 2019

Autosomal recessive polycystic kidney disease (ARPKD) is a severe pediatric hepatorenal disorder with pronounced phenotypic variability. A substantial number of patients with early diagnosis reaches adulthood and some patients are not diagnosed until adulthood. Yet, clinical knowledge about adult ARPKD patients is scarce. Here, we describe forty-nine patients with longitudinal follow-up into young adulthood that were identified in the international ARPKD cohort study ARegPKD. Forty-five patients were evaluated in a cross-sectional analysis at a mean age of 21.4 (±3.3) years describing hepatorenal findings. Renal function of native kidneys was within CKD stages 1 to 3 in more than 50% of the patients. Symptoms of hepatic involvement were frequently detected. Fourteen (31%) patients had undergone kidney transplantation and six patients (13%) had undergone liver transplantation or combined liver and kidney transplantation prior to the visit revealing a wide variability of clinical courses. Hepatorenal involvement and preceding complications in other organs were also evaluated in a time-to-event analysis. In summary, we characterize the broad clinical spectrum of young adult ARPKD patients. Importantly, many patients have a stable renal and hepatic situation in young adulthood. ARPKD should also be considered as a differential diagnosis in young adults with fibrocystic hepatorenal disease.
Novel Genes and Metabolite Trends in Bifidobacterium longum subsp. infantis Bi-26 Metabolism of Human Milk Oligosaccharide 2′-fucosyllactose
Authors: Zabel Bryan , Yde Christian Clement , Roos Paige , Marcussen Jørn , Jensen Henrik Max , Salli Krista , Hirvonen Johanna , Ouwehand Arthur C. , Morovic Wesley
Published: May 2019

Human milk oligosaccharides (HMOs) function as prebiotics for beneficial bacteria in the developing gut, often dominated by Bifidobacterium spp. To understand the relationship between bifidobacteria utilizing HMOs and how the metabolites that are produced could affect the host, we analyzed the metabolism of HMO 2′-fucosyllactose (2′-FL) in Bifidobacterium longum subsp. infantis Bi-26. RNA-seq and metabolite analysis (NMR/GCMS) was performed on samples at early (A600 = 0.25), mid-log (0.5–0.7) and late-log phases (1.0–2.0) of growth. Transcriptomic analysis revealed many gene clusters including three novel ABC-type sugar transport clusters to be upregulated in Bi-26 involved in processing of 2′-FL along with metabolism of its monomers glucose, fucose and galactose. Metabolite data confirmed the production of formate, acetate, 1,2-propanediol, lactate and cleaving of fucose from 2′-FL. The formation of acetate, formate, and lactate showed how the cell uses metabolites during fermentation to produce higher levels of ATP (mid-log compared to other stages) or generate cofactors to balance redox. We concluded that 2′-FL metabolism is a complex process involving multiple gene clusters, that produce a more diverse metabolite profile compared to lactose. These results provide valuable insight on the mode-of-action of 2′-FL utilization by Bifidobacterium longum subsp. infantis Bi-26.
The role of stratospheric ozone for Arctic-midlatitude linkages
Authors: Romanowsky Erik , Handorf Dörthe , Jaiser Ralf , Wohltmann Ingo , Dorn Wolfgang , Ukita Jinro , Cohen Judah , Dethloff Klaus , Rex Markus
Published: May 2019

Arctic warming was more pronounced than warming in midlatitudes in the last decades making this region a hotspot of climate change. Associated with this, a rapid decline of sea-ice extent and a decrease of its thickness has been observed. Sea-ice retreat allows for an increased transport of heat and momentum from the ocean up to the tropo- and stratosphere by enhanced upward propagation of planetary-scale atmospheric waves. In the upper atmosphere, these waves deposit the momentum transported, disturbing the stratospheric polar vortex, which can lead to a breakdown of this circulation with the potential to also significantly impact the troposphere in mid- to late-winter and early spring. Therefore, an accurate representation of stratospheric processes in climate models is necessary to improve the understanding of the impact of retreating sea ice on the atmospheric circulation. By modeling the atmospheric response to a prescribed decline in Arctic sea ice, we show that including interactive stratospheric ozone chemistry in atmospheric model calculations leads to an improvement in tropo-stratospheric interactions compared to simulations without interactive chemistry. This suggests that stratospheric ozone chemistry is important for the understanding of sea ice related impacts on atmospheric dynamics.
Gut microbiome differences between wild and captive black rhinoceros – implications for rhino health
Authors: Gibson Keylie M. , Nguyen Bryan N. , Neumann Laura M. , Miller Michele , Buss Peter , Daniels Savel , Ahn Michelle J. , Crandall Keith A. , Pukazhenthi Budhan
Published: May 2019

A number of recent studies have shown the importance of the mammalian gut microbiome in host health. In the context of endangered species, a few studies have examined the relationship between the gut microbiome in wild versus captive populations due to digestive and other health issues. Unfortunately, the results seem to vary across taxa in terms of captive animals having higher, lower, or equivalent microbiome diversity relative to their wild counterparts. Here, we focus on the black rhinoceros as captive animals suffer from a number of potentially dietary related health effects. We compared gut microbiomes of wild and captive black rhinos to test for differences in taxonomic diversity (alpha and beta) and in functional diversity of the microbiome. We incorporated a more powerful metagenomic shotgun sequencing approach rather than a targeted amplification of the 16S gene for taxonomic assignment of the microbiome. Our results showed no significant differences in the alpha diversity levels between wild and captive black rhinos, but significant differences in beta diversity. We found that bacterial taxa traditionally associated with ruminant guts of domesticated animals had higher relative abundances in captive rhinos. Our metagenomic sequencing results suggest that unknown gut microbes of wild rhinos are being replaced by those found in conventional human-domesticated livestock. Wild rhinos have significantly different functional bacterial communities compared to their captive counterparts. Functional profiling results showed greater abundance of glycolysis and amino acid synthesis pathways in captive rhino microbiomes, representing an animal receiving sub-optimal nutrition with a readily available source of glucose but possibly an imbalance of necessary macro and micronutrients. Given the differences observed between wild and captive rhino gut microbiomes, we make a number of recommendations for potentially modifying captive gut microbiome to better reflect their wild counterparts and thereby hopefully improve overall rhino health in captivity.
A hydrofluoric acid-free method to dissolve and quantify silica nanoparticles in aqueous and solid matrices
Authors: Bossert David , Urban Dominic A. , Maceroni Mattia , Ackermann-Hirschi Liliane , Haeni Laetitia , Yajan Phattadon , Spuch-Calvar Miguel , Rothen-Rutishauser Barbara , Rodriguez-Lorenzo Laura , Petri-Fink Alke , Schwab Fabienne
Published: May 2019

As the commercial use of synthetic amorphous silica nanomaterials (SiO2-NPs) increases, their effects on the environment and human health have still not been explored in detail. An often-insurmountable obstacle for SiO2-NP fate and hazard research is the challenging analytics of solid particulate silica species, which involves toxic and corrosive hydrofluoric acid (HF). We therefore developed and validated a set of simple hydrofluoric acid-free sample preparation methods for the quantification of amorphous SiO2 micro- and nanoparticles. To circumvent HF, we dissolved the SiO2-NPs by base-catalyzed hydrolysis at room temperature or under microwave irradiation using potassium hydroxide, replacing the stabilizing fluoride ions with OH−, and exploiting the stability of the orthosilicic acid monomer under a strongly basic pH. Inductively coupled plasma – optical emission spectroscopy (ICP-OES) or a colorimetric assay served to quantify silicon. The lowest KOH: SiO2 molar ratio to effectively dissolve and quantify SiO2-NPs was 1.2 for colloidal Stöber SiO2-NPs at a pH >12. Fumed SiO2-NPs (Aerosil®) or food grade SiO2 (E551) containing SiO2-NPs were degradable at higher KOH: SiO2 ratios >8000. Thus, hydrofluoric acid-free SiO2-NP digestion protocols based on KOH present an effective (recoveries of >84%), less hazardous, and easy to implement alternative to current methods.
Wolves contribute to disease control in a multi-host system
Authors: Tanner E. , White A. , Acevedo P. , Balseiro A. , Marcos J. , Gortázar C.
Published: May 2019

We combine model results with field data for a system of wolves (Canis lupus) that prey on wild boar (Sus scrofa), a wildlife reservoir of tuberculosis, to examine how predation may contribute to disease control in multi-host systems. Results show that predation can lead to a marked reduction in the prevalence of infection without leading to a reduction in host population density since mortality due to predation can be compensated by a reduction in disease induced mortality. A key finding therefore is that a population that harbours a virulent infection can be regulated at a similar density by disease at high prevalence or by predation at low prevalence. Predators may therefore provide a key ecosystem service which should be recognised when considering human-carnivore conflicts and the conservation and re-establishment of carnivore populations.
Cynomolgus macaque IL37 polymorphism and control of SIV infection
Authors: Shiina Takashi , Suzuki Shingo , Congy-Jolivet Nicolas , Aarnink Alice , Garchon Henri-Jean , Dereuddre-Bosquet Nathalie , Vaslin Bruno , Tchitchek Nicolas , Desjardins Delphine , Autran Brigitte , Lambotte Olivier , Theodorou Ioannis , Le Grand Roger , Blancher Antoine
Published: May 2019

The association between gene polymorphisms and plasma virus load at the set point (SP-PVL) was investigated in Mauritian macaques inoculated with SIV. Among 44 macaques inoculated with 50 AID50, six individuals were selected: three with SP-PVL among the highest and three with SP-PVL among the lowest. The exons of 390 candidate genes of these six animals were sequenced. Twelve non-synonymous single nucleotide polymorphisms (NS-SNPs) lying in nine genes potentially associated with PVL were genotyped in 23 animals. Three NS-SNPs with probabilities of association with PVL less than 0.05 were genotyped in a total of 44 animals. One NS-SNP lying in exon 1 of the IL37 gene displayed a significant association (p = 3.33 × 10−4) and a strong odds ratio (19.52). Multiple linear regression modeling revealed three significant predictors of SP-PVL, including the IL37 exon 1 NS-SNP (p = 0.0004) and the MHC Class IB haplotypes M2 (p = 0.0007) and M6 (p = 0.0013). These three factors in conjunction explained 48% of the PVL variance (p = 4.8 × 10−6). The potential role of IL37 in the control of SIV infection is discussed.
Early exhumation of the Frontal Cordillera (Southern Central Andes) and implications for Andean mountain-building at ~33.5°S
Authors: Riesner Magali , Simoes Martine , Carrizo Daniel , Lacassin Robin
Published: May 2019

The Andes are the modern active example of a Cordilleran-type orogen, with mountain-building and crustal thickening within the upper plate of a subduction zone. Despite numerous studies of this emblematic mountain range, several primary traits of this orogeny remain unresolved or poorly documented. The onset of uplift and deformation of the Frontal Cordillera basement culmination of the Southern Central Andes is such an example, even though this structural unit appears as a first-order topographic and geological feature. To solve for this, new (U-Th)/He ages on apatite and zircon from granitoids of the Frontal Cordillera at ~33.5°S are provided here. These data, interpreted as an age-elevation thermochronological profile, imply continuous exhumation initiating well before ~12–14 Ma, and at most by ~22 Ma when considering the youngest zircon grain from the lowermost sample. The age of exhumation onset is then refined to ~20 Ma by combining these results with data on sedimentary provenance from the nearby basins. Such continuous exhumation since ~20 Ma needs to have been sustained by tectonic uplift on an underlying crustal-scale thrust ramp. Such early exhumation and associated uplift of the Frontal Cordillera invalidate the classically proposed east-vergent models of the Andes at this latitude. Additionally, they provide further support to recent views on Andean mountain-building proposing that the Andes at ~33.5°S grew firstly over west-vergent basement structures.
Sperm motility of oysters from distinct populations differs in response to ocean acidification and freshening
Authors: Falkenberg Laura J. , Styan Craig A. , Havenhand Jon N.
Published: May 2019

Species’ responses to climate change will reflect variability in the effects of physiological selection that future conditions impose. Here, we considered the effects of ocean acidification (increases in pCO2; 606, 925, 1250 µatm) and freshening (reductions in salinity; 33, 23, 13 PSU) on sperm motility in oysters (Crassostrea gigas) from two populations (one recently invaded, one established for 60+ years). Freshening reduced sperm motility in the established population, but this was offset by a positive effect of acidification. Freshening also reduced sperm motility in the recently invaded population, but acidification had no effect. Response direction, strength, and variance differed among individuals within each population. For the established population, freshening increased variance in sperm motility, and exposure to both acidification and freshening modified the performance rank of males (i.e. rank motility of sperm). In contrast, for the recently invaded population, freshening caused a smaller change in variance, and male performance rank was broadly consistent across treatments. That inter-population differences in response may be related to environmental history (recently invaded, or established), indicates this could influence scope for selection and adaptation. These results highlight the need to consider variation within and among population responses to forecast effects of multiple environmental change drivers.
Multimodal Wrist Biosensor for Wearable Cuff-less Blood Pressure Monitoring System
Authors: Rachim Vega Pradana , Chung Wan-Young
Published: May 2019

We propose a multimodal biosensor for use in continuous blood pressure (BP) monitoring system. Our proposed novel configuration measures photo-plethysmography (PPG) and impedance plethysmography (IPG) signals simultaneously from the subject wrist. The proposed biosensor system enables a fully non-intrusive system that is cuff-less, also utilize a single measurement site for maximum wearability and convenience of the patients. The efficacy of the proposed technique was evaluated on 10 young healthy subjects. Experimental results indicate that the pulse transit time (PTT)-based features calculated from an IPG peak and PPG maximum second derivative (f14) had a relatively high correlation coefficient (r) to the reference BP, with −0.81 ± 0.08 and −0.78 ± 0.09 for systolic BP (SBP) and diastolic BP (DBP), respectively. Moreover, here we proposed two BP estimation models that utilize six- and one-point calibration models. The six-point model is based on the PTT, whereas the one-point model is based on the combined PTT and radial impedance (Z). Thus, in both models, we observed an adequate root-mean-square-error estimation performance, with 4.20 ± 1.66 and 2.90 ± 0.90 for SBP and DBP, respectively, with the PTT BP model; and 6.86 ± 1.65 and 6.67 ± 1.75 for SBP and DBP, respectively, with the PTT-Z BP model. This study suggests the possibility of estimating a subject’s BP from only wrist bio-signals. Thus, the six- and one-point PTT-Z calibration models offer adequate performance for practical applications.
RETRACTED ARTICLE: The Adipocyte Na/K-ATPase Oxidant Amplification Loop is the Central Regulator of Western Diet-Induced Obesity and Associated Comorbidities
Authors: Pratt Rebecca D. , Brickman Cameron , Nawab Athar , Cottrill Cameron , Snoad Brian , Lakhani Hari Vishal , Jelcick Austin , Henderson Brandon , Bhardwaj Niharika N. , Sanabria Juan R. , Liu Jiang , Xie Zijian , Abraham Nader G. , Shapiro Joseph I. , Sodhi Komal
Published: May 2019

Obesity has become a worldwide epidemic. We have previously reported that systemic administration of pNaKtide which targets the Na/K-ATPase oxidant amplification loop (NKAL) was able to decrease systemic oxidative stress and adiposity in mice fed a high fat and fructose supplemented western diet (WD). As adipocytes are believed to play a central role in the development of obesity and its related comorbidities, we examined whether lentiviral-mediated adipocyte-specific expression of NaKtide, a peptide derived from the N domain of the alpha1 Na/K-ATPase subunit, could ameliorate the effects of the WD. C57BL6 mice were fed a WD, which activated Na/K-ATPase signaling in the adipocytes and induced an obese phenotype and caused an increase in plasma levels of leptin, IL-6 and TNFα. WD also decreased locomotor activity, expression of the D2 receptor and tyrosine hydroxylase in brain tissue, while markers of neurodegeneration and neuronal apoptosis were increased following the WD. Selective adipocyte expression of NaKtide in these mice fed a WD attenuated all of these changes including the brain biochemical alterations and behavioral adaptations. These data suggest that adipocyte derived cytokines play an essential role in the development of obesity induced by a WD and that targeting the adipocyte NKAL loop may serve as an effective therapeutic strategy.
An electrochemiluminescence based assay for quantitative detection of endogenous and exogenously applied MeCP2 protein variants
Authors: Steinkellner Hannes , Schönegger Anna , Etzler Julia , Kempaiah Prakasha , Huber Anna , Hahn Kathrin , Rose Katrin , Duerr Mark , Christodoulou John , Beribisky Alexander V. , Neuhaus Winfried , Laccone Franco
Published: May 2019

Methyl-CpG-binding protein 2 (MeCP2) is a multifunctional chromosomal protein that plays a key role in the central nervous system. Its levels need to be tightly regulated, as both deficiency and excess of the protein can lead to severe neuronal dysfunction. Loss-of-function mutations affecting MeCP2 are the primary cause of Rett syndrome (RTT), a severe neurological disorder that is thought to result from absence of functional protein in the brain. Several therapeutic strategies for the treatment of RTT are currently being developed. One of them is the use of stable and native TAT-MeCP2 fusion proteins to replenish its levels in neurons after permeation across the blood-brain barrier (BBB). Here we describe the expression and purification of various transactivator of transcription (TAT)-MeCP2 variants and the development of an electrochemiluminescence based assay (ECLIA) that is able to measure endogenous MeCP2 and recombinant TAT-MeCP2 fusion protein levels in a 96-well plate format. The MeCP2 ECLIA produces highly quantitative, accurate and reproducible measurements with low intra- and inter-assay error throughout a wide working range. To underline its broad applicability, this assay was used to analyze brain tissue and study the transport of TAT-MeCP2 variants across an in vitro model of the blood-brain barrier.
A structural and biochemical comparison of Ribonuclease E homologues from pathogenic bacteria highlights species-specific properties
Authors: Mardle Charlotte E. , Shakespeare Thomas J. , Butt Louise E. , Goddard Layla R. , Gowers Darren M. , Atkins Helen S. , Vincent Helen A. , Callaghan Anastasia J.
Published: May 2019

Regulation of gene expression through processing and turnover of RNA is a key mechanism that allows bacteria to rapidly adapt to changing environmental conditions. Consequently, RNA degrading enzymes (ribonucleases; RNases) such as the endoribonuclease RNase E, frequently play critical roles in pathogenic bacterial virulence and are potential antibacterial targets. RNase E consists of a highly conserved catalytic domain and a variable non-catalytic domain that functions as the structural scaffold for the multienzyme degradosome complex. Despite conservation of the catalytic domain, a recent study identified differences in the response of RNase E homologues from different species to the same inhibitory compound(s). While RNase E from Escherichia coli has been well-characterised, far less is known about RNase E homologues from other bacterial species. In this study, we structurally and biochemically characterise the RNase E catalytic domains from four pathogenic bacteria: Yersinia pestis, Francisella tularensis, Burkholderia pseudomallei and Acinetobacter baumannii, with a view to exploiting RNase E as an antibacterial target. Bioinformatics, small-angle x-ray scattering and biochemical RNA cleavage assays reveal globally similar structural and catalytic properties. Surprisingly, subtle species-specific differences in both structure and substrate specificity were also identified that may be important for the development of effective antibacterial drugs targeting RNase E.
Cytotoxicity of trifluridine correlates with the thymidine kinase 1 expression level
Authors: Kataoka Yuki , Iimori Makoto , Niimi Shinichiro , Tsukihara Hiroshi , Wakasa Takeshi , Saeki Hiroshi , Oki Eiji , Maehara Yoshihiko , Kitao Hiroyuki
Published: May 2019

Trifluridine (FTD), a tri-fluorinated thymidine analogue, is a key component of the oral antitumor drug FTD/TPI (also known as TAS-102), which is used to treat refractory metastatic colorectal cancer. Thymidine kinase 1 (TK1) is thought to be important for the incorporation of FTD into DNA, resulting in DNA dysfunction and cytotoxicity. However, it remains unknown whether TK1 is essential for FTD incorporation into DNA and whether this event is affected by the expression level of TK1 because TK1-specific-deficient human cancer cell lines have not been established. Here, we generated TK1-knock-out human colorectal cancer cells using the CRISPR/Cas9 genome editing system and validated the specificity of TK1 knock-out by measuring expression of AFMID, which is encoded on the same locus as TK1. Using TK1-knock-out cells, we confirmed that TK1 is essential for cellular sensitivity to FTD. Furthermore, we demonstrated a correlation between the TK1 expression level and cytotoxicity of FTD using cells with inducible TK1 expression, which were generated from TK1-knock-out cells. Based on our finding that the TK1 expression level correlates with sensitivity to FTD, we suggest that FTD/TPI might efficiently treat cancers with high TK1 expression.
New PCR primers for metabarcoding environmental DNA from freshwater eels, genus Anguilla
Authors: Takeuchi Aya , Sado Tetsuya , Gotoh Ryo O. , Watanabe Shun , Tsukamoto Katsumi , Miya Masaki
Published: May 2019

Freshwater eels of the genus Anguilla comprise 16 species that include three subspecies and are characterized by their unique catadromous life cycles. Their life histories and nocturnal life styles make it difficult to observe them in freshwater and marine habitats. To investigate their distribution and ecology in aquatic environments, we developed new PCR primers for metabarcoding environmental DNA (eDNA) from Anguilla. The new primers (MiEel) were designed for two conserved regions of the mitochondrial ATP6 gene, which amplify a variable region with sufficient interspecific variations ranging from five to 22 nucleotide differences (one to three nucleotide differences between three subspecies pairs). We confirmed the versatility of the MiEel primers for all freshwater eels using tissue DNA extracts when analyzed separately. The metabarcoding combined with the MiEel primers using mock communities enabled simultaneous detection of Anguilla at the species level. Analysis of eDNA samples from aquarium tanks, a controlled pond and natural rivers demonstrated that the MiEel metabarcoding could successfully detect the correct Anguilla species from water samples. These results suggested that eDNA metabarcoding with MiEel primers would be useful for non-invasively monitoring the presence of the endangered anguillid eels in aquatic environments where sampling surveys are difficult.
Comparative genomics approach to build a genome-wide database of high-quality, informative microsatellite markers: application on Phytophthora sojae, a soybean pathogen
Authors: Cai Guohong , Fleury Tomara J. , Zhang Ning
Published: May 2019

Microsatellites are a tract of repetitive, short DNA motifs (usually 1 to 6 bp) abundant in eukaryotic genomes. They are robust molecular markers in many areas of studies. Development of microsatellite markers usually involves three steps: (1) obtaining microsatellite-containing sequences, (2) primer design, and (3) screening microsatellite loci for polymorphism. The first and third steps require considerable resources. Next generation sequencing technologies have greatly alleviated the constraint of the first step. In this study, we leveraged the availability of genome assemblies of multiple individuals in many species and designed a comparative genomics approach to bioinformatically identify polymorphic loci. Our approach can eliminate or greatly reduce the need of experimental screening for polymorphism and ensure that the flanking regions do not have length difference that would confound interpretation of genotyping results using microsatellite markers. We applied this approach onPhytophthora sojae, a soybean pathogen, and identified 157 high-quality, informative microsatellite markers in this oomycete. Experimental validation of 20 loci supported bioinformatics predictions. Our approach can be readily applied to other organisms of which the genomes of multiple individuals have been sequenced.
RNA-seq analysis and fluorescence imaging of melon powdery mildew disease reveal an orchestrated reprogramming of host physiology
Authors: Polonio Álvaro , Pineda Mónica , Bautista Rocío , Martínez-Cruz Jesús , Pérez-Bueno María Luisa , Barón Matilde , Pérez-García Alejandro
Published: May 2019

The cucurbit powdery mildew elicited by Podosphaera xanthii is one of the most important limiting factors in cucurbit production. Our knowledge of the genetic and molecular bases underlying the physiological processes governing this disease is very limited. We used RNA-sequencing to identify differentially expressed genes in leaves of Cucumis melo upon inoculation with P. xanthii, using RNA samples obtained at different time points during the early stages of infection and their corresponding uninfected controls. In parallel, melon plants were phenotypically characterized using imaging techniques. We found a high number of differentially expressed genes (DEGs) in infected plants, which allowed for the identification of many plant processes that were dysregulated by the infection. Among those, genes involved in photosynthesis and related processes were found to be upregulated, whereas genes involved in secondary metabolism pathways, such as phenylpropanoid biosynthesis, were downregulated. These changes in gene expression could be functionally validated by chlorophyll fluorescence imaging and blue-green fluorescence imaging analyses, which corroborated the alterations in photosynthetic activity and the suppression of phenolic compound biosynthesis. The powdery mildew disease in melon is a consequence of a complex and multifaceted process that involves the dysregulation of many plant pathways such as primary and secondary metabolism.
Deciphering host lysosome-mediated elimination of Plasmodium berghei liver stage parasites
Authors: Niklaus L. , Agop-Nersesian C. , Schmuckli-Maurer J. , Wacker R. , Grünig V. , Heussler V. T.
Published: May 2019

Liver stage Plasmodium parasites reside in a parasitophorous vacuole (PV) that associates with lysosomes. It has previously been shown that these organelles can have beneficial as well as harmful effects on the parasite. Yet it is not clear how the association of lysosomes with the parasite is controlled and how interactions with these organelles lead to the antagonistic outcomes. In this study we used advanced imaging techniques to characterize lysosomal interactions with the PV. In host cells harboring successfully developing parasites we observed that these interaction events reach an equilibrium at the PV membrane (PVM). In a population of arrested parasites, this equilibrium appeared to shift towards a strongly increased lysosomal fusion with the PVM witnessed by strong PVM labeling with the lysosomal marker protein LAMP1. This was followed by acidification of the PV and elimination of the parasite. To systematically investigate elimination of arrested parasites, we generated transgenic parasites that express the photosensitizer KillerRed, which leads to parasite killing after activation. Our work provides insights in cellular details of intracellular killing and lysosomal elimination of Plasmodium parasites independent of cells of the immune system.
Encapsulation of a TRPM8 Agonist, WS12, in Lipid Nanocapsules Potentiates PC3 Prostate Cancer Cell Migration Inhibition through Channel Activation
Authors: Grolez G. P. , Hammadi M. , Barras A. , Gordienko D. , Slomianny C. , Völkel P. , Angrand P. O. , Pinault M. , Guimaraes C. , Potier-Cartereau M. , Prevarskaya N. , Boukherroub R. , Gkika D.
Published: May 2019

In prostate carcinogenesis, expression and/or activation of the Transient Receptor Potential Melastatin 8 channel (TRPM8) was shown to block in vitro Prostate Cancer (PCa) cell migration. Because of their localization at the plasma membrane, ion channels, such as TRPM8 and other membrane receptors, are promising pharmacological targets. The aim of this study was thus to use nanocarriers encapsulating a TRPM8 agonist to efficiently activate the channel and therefore arrest PCa cell migration. To achieve this goal, the most efficient TRPM8 agonist, WS12, was encapsulated into Lipid NanoCapsules (LNC). The effect of the nanocarriers on channel activity and cellular physiological processes, such as cell viability and migration, were evaluated in vitro and in vivo. These results provide a proof-of-concept support for using TRPM8 channel-targeting nanotechnologies based on LNC to develop more effective methods inhibiting PCa cell migration in zebrafish xenograft.
Stable and manipulable Bloch point
Authors: Beg Marijan , Pepper Ryan A. , Cortés-Ortuño David , Atie Bilal , Bisotti Marc-Antonio , Downing Gary , Kluyver Thomas , Hovorka Ondrej , Fangohr Hans
Published: May 2019

The prediction of magnetic skyrmions being used to change the way we store and process data has led to materials with Dzyaloshinskii-Moriya interaction coming into the focus of intensive research. So far, studies have looked mostly at magnetic systems composed of materials with single chirality. In a search for potential future spintronic devices, combination of materials with different chirality into a single system may represent an important new avenue for research. Using finite element micromagnetic simulations, we study an FeGe disk with two layers of different chirality. We show that for particular thicknesses of layers, a stable Bloch point emerges at the interface between two layers. In addition, we demonstrate that the system undergoes hysteretic behaviour and that two different types of Bloch point exist. These ‘head-to-head’ and ‘tail-to-tail’ Bloch point configurations can, with the application of an external magnetic field, be switched between. Finally, by investigating the time evolution of the magnetisation field, we reveal the creation mechanism of the Bloch point. Our results introduce a stable and manipulable Bloch point to the collection of particle-like state candidates for the development of future spintronic devices.