bluuebunny/crossref_metadata_2025 huggingface dataset:
Astrocytic endfeet re-cover blood vessels after removal by laser ablation
Authors: Kubotera Hideaki , Ikeshima-Kataoka Hiroko , Hatashita Yoshiki , Allegra Mascaro Anna Letizia , Pavone Francesco Saverio , Inoue Takafumi
Published: February 2019

The astrocyte, one of the glial cells, plays many functional roles. These include provision of nutrients from blood vessels to neurons, supply of neurotransmitters and support of blood–brain barrier (BBB) integrity. Astrocytes are known to support the integrity of BBB through maintenance of the tight junction between endothelial cells of blood vessels. However, evidence of its direct contribution to BBB is lacking owing to technical limitations. In this study, astrocytic endfeet covering blood vessels were removed by the laser ablation method with two photon laser scanning microscopy in in vivo mouse brain, and the re-covering of blood vessels with the astrocytic endfeet was observed in about half of the cases. Blood vessels kept their integrity without astrocytic endfoot covers: leakage of plasma marker dyes, Evans Blue or dextran-conjugated fluorescein, was not observed from stripped blood vessels, while ablation of vascular walls induced extravasation of Evans Blue. These results suggest that the astrocytic endfeet covering blood vessels do not contribute to the immediate BBB barrier.
A cancer-associated point mutation disables the steric gate of human PrimPol
Authors: Díaz-Talavera Alberto , Calvo Patricia A. , González-Acosta Daniel , Díaz Marcos , Sastre-Moreno Guillermo , Blanco-Franco Luis , Guerra Susana , Martínez-Jiménez Maria I. , Méndez Juan , Blanco Luis
Published: February 2019

PrimPol is a human primase/polymerase specialized in re-starting stalled forks by repriming beyond lesions such as pyrimidine dimers, and replication-perturbing structures including G-quadruplexes and R-loops. Unlike most conventional primases, PrimPol proficiently discriminates against ribonucleotides (NTPs), being able to start synthesis using deoxynucleotides (dNTPs), yet the structural basis and physiological implications for this discrimination are not understood. In silico analyses based on the three-dimensional structure of human PrimPol and related enzymes enabled us to predict a single residue, Tyr100, as the main effector of sugar discrimination in human PrimPol and a change of Tyr100 to histidine to boost the efficiency of NTP incorporation. We show here that the Y100H mutation profoundly stimulates NTP incorporation by human PrimPol, with an efficiency similar to that for dNTP incorporation during both primase and polymerase reactions in vitro. As expected from the higher cellular concentration of NTPs relative to dNTPs, Y100H expression in mouse embryonic fibroblasts and U2OS osteosarcoma cells caused enhanced resistance to hydroxyurea, which decreases the dNTP pool levels in S-phase. Remarkably, the Y100H PrimPol mutation has been identified in cancer, suggesting that this mutation could be selected to promote survival at early stages of tumorigenesis, which is characterized by depleted dNTP pools.
Structural biology workflow for the expression and characterization of functional human sodium glucose transporter type 1 in Pichia pastoris
Authors: Suades Albert , Alcaraz Antonio , Cruz Esteban , Álvarez-Marimon Elena , Whitelegge Julian P. , Manyosa Joan , Cladera Josep , Perálvarez-Marín Alex
Published: February 2019

Heterologous expression of human membrane proteins is a challenge in structural biology towards drug discovery. Here we report a complete expression and purification process of a functional human sodium/D-glucose co-transporter 1 (hSGLT1) in Pichia pastoris as representative example of a useful strategy for any human membrane protein. hSGLT1 gene was cloned in two different plasmids to develop parallel strategies: one which includes green fluorescent protein fusion for screening optimal conditions, and another for large scale protein production for structural biology and biophysics studies. Our strategy yields at least 1 mg of monodisperse purified recombinant hSGLT1 per liter of culture, which can be characterized by circular dichroism and infrared spectroscopy as an alpha-helical fold protein. This purified hSGLT1 transports co-substrates (Na+ and glucose) and it is inhibited by phlorizin in electrophysiological experiments performed in planar lipid membranes.
Contribution of syndecans to cellular internalization and fibrillation of amyloid-β(1–42)
Authors: Letoha Tamás , Hudák Anett , Kusz Erzsébet , Pettkó-Szandtner Aladár , Domonkos Ildikó , Jósvay Katalin , Hofmann-Apitius Martin , Szilák László
Published: February 2019

Intraneuronal accumulation of amyloid-β(1–42) (Aβ1–42) is one of the earliest signs of Alzheimer’s disease (AD). Cell surface heparan sulfate proteoglycans (HSPGs) have profound influence on the cellular uptake of Aβ1–42 by mediating its attachment and subsequent internalization into the cells. Colocalization of amyloid plaques with members of the syndecan family of HSPGs, along with the increased expression of syndecan-3 and -4 have already been reported in postmortem AD brains. Considering the growing evidence on the involvement of syndecans in the pathogenesis of AD, we analyzed the contribution of syndecans to cellular uptake and fibrillation of Aβ1–42. Among syndecans, the neuron specific syndecan-3 isoform increased cellular uptake of Aβ1–42 the most. Kinetics of Aβ1–42 uptake also proved to be fairly different among SDC family members: syndecan-3 increased Aβ1–42 uptake from the earliest time points, while other syndecans facilitated Aβ1–42 internalization at a slower pace. Internalized Aβ1–42 colocalized with syndecans and flotillins, highlighting the role of lipid-rafts in syndecan-mediated uptake. Syndecan-3 and 4 also triggered fibrillation of Aβ1–42, further emphasizing the pathophysiological relevance of syndecans in plaque formation. Overall our data highlight syndecans, especially the neuron-specific syndecan-3 isoform, as important players in amyloid pathology and show that syndecans, regardless of cell type, facilitate key molecular events in neurodegeneration.
Mechanism of action of the moonlighting protein EfTu as a Substance P sensor in Bacillus cereus
Authors: N’Diaye Awa R. , Borrel Valerie , Racine Pierre-Jean , Clamens Thomas , Depayras Segolene , Maillot Olivier , Schaack Beatrice , Chevalier Sylvie , Lesouhaitier Olivier , Feuilloley Marc G. J.
Published: February 2019

The striking feature of the ubiquitous protein EfTu (Thermo unstable ribosomal Elongation factor) is its moonlighting (multifunctional) activity. Beyond its function at the ribosomal level it should be exported to the bacterial surface and act as an environmental sensor. In Bacillus cereus, and other cutaneous bacteria, it serves as a Substance P (SP) receptor and is essential for bacterial adaptation to the host. However, the modus operandi of EfTu as a bacterial sensor remains to be investigated. Studies realized by confocal and transmission electron microscopy revealed that, in the absence of an exogenous signal, EfTu is not exposed on the bacterial surface but is recruited under the effect of SP. In addition, SP acts as a transcriptional regulator of the tuf gene encoding for EfTu. As observed using gadolinium chloride, an inhibitor of membrane mechanosensitive channels (Msc), Msc control EfTu export and subsequently the bacterial response to SP both in terms of cytotoxicity and biofilm formation activity. Microscale thermophoresis revealed that in response to SP, EfTu can form homopolymers. This event should occur after EfTu export and, as shown by proteo-liposome reconstruction studies, SP appears to promote EfTu polymers association to the membrane, leading subsequently to the bacterial response. Molecular modeling suggests that this mechanism should involve EfTu unfolding and insertion into the bacterial cytoplasmic membrane, presumably through formation of homopolymers. This study is unraveling the original mechanism action of EfTu as a bacterial sensor but also reveals that this protein should have a broader role, including in eukaryotes.
Laser- synthesized TiN nanoparticles as promising plasmonic alternative for biomedical applications
Authors: Popov Anton A. , Tselikov Gleb , Dumas Noé , Berard Charlotte , Metwally Khaled , Jones Nicola , Al-Kattan Ahmed , Larrat Benoit , Braguer Diane , Mensah Serge , Da Silva Anabela , Estève Marie-Anne , Kabashin Andrei V.
Published: February 2019

Exhibiting a red-shifted absorption/scattering feature compared to conventional plasmonic metals, titanium nitride nanoparticles (TiN NPs) look as very promising candidates for biomedical applications, but these applications are still underexplored despite the presence of extensive data for conventional plasmonic counterparts. Here, we report the fabrication of ultrapure, size-tunable TiN NPs by methods of femtosecond laser ablation in liquids and their biological testing. We show that TiN NPs demonstrate strong and broad plasmonic peak around 640–700 nm with a significant tail over 800 nm even for small NPs sizes (<7 nm). In vitro tests of laser-synthesized TiN NPs on cellular models evidence their low cytotoxicity and excellent cell uptake. We finally demonstrate a strong photothermal therapy effect on U87–MG cancer cell cultures using TiN NPs as sensitizers of local hyperthermia under near-infrared laser excitation. Based on absorption band in the region of relative tissue transparency and acceptable biocompatibility, laser-synthesized TiN NPs promise the advancement of biomedical modalities employing plasmonic effects, including absorption/scattering contrast imaging, photothermal therapy, photoacoustic imaging and SERS.
High-velocity impact of solid objects on Non-Newtonian Fluids
Authors: de Goede Thijs C. , de Bruin Karla G. , Bonn Daniel
Published: February 2019

We investigate which property of non-Newtonian fluids determines the deceleration of a high-speed impacting object. Using high-speed camera footage, we measure the velocity decrease of a high-speed spherical object impacting a typical Newtonian fluid (water) as a reference and compare it with a shear thickening fluid (cornstarch) and a shear thinning viscoelastic fluid (a weakly cross-linked polymer gel). Three models describing the kinetic energy loss of the object are considered: fluid inertia, shear thickening and viscoelasticity. By fitting the three models to the experimental data, we conclude that the viscoelastic model works best for both the cornstarch and the polymer gel. Since the cornstarch is also viscoelastic, we conclude that the ability to stop objects of these complex fluids is given by their viscoelasticity rather than shear thickening or shear thinning.
An ADAMTS Sol narae is required for cell survival in Drosophila
Authors: Tsogtbaatar Orkhon , Won Jong-Hoon , Kim Go-Woon , Han Jeong-Hoon , Bae Young-Kyung , Cho Kyung-Ok
Published: February 2019

Cell survival is essential for all living organisms to cope against multiple environmental insults. Intercellular signaling between dying and surviving cells plays an important role to ensure compensatory proliferation, preventing tissue loss after environmental stresses. Here, we show that Sol narae (Sona), a Disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) in Drosophila is required for cell survival. sona exhibited a positive genetic interaction with Death-associated inhibitor of apoptosis 1 (Diap1), and a negative genetic interaction with reaper (rpr). Transcription patterns of sona, Diap1, and rpr genes in the pouch region of wing discs were coordinately changed after irradiation. Interestingly, there was a negative correlation in the expression levels of Sona and DIAP1, and both cell types, one with high Sona level and the other with high Diap1 level, were resistant to irradiation-induced cell death. The sona-expressing cells rarely entered into cell cycle themselves but promoted the nearby cells to proliferate in irradiation conditions. We found that these sona-expressing cells are able to upregulate Cyclin D (Cyc D) and increase tissue size. Furthermore, transient Sona overexpression increased survival rate and promoted development of flies in irradiation conditions. We propose that the two types of radiation-resistant cells, one with high Sona level and the other with high Diap1 level, communicate with dying cells and between each other for cell survival and proliferation in response to irradiation.
Ultra-High Efficiency and Broad Band Operation of Infrared Metasurface Anomalous Reflector based on Graphene Plasmonics
Authors: Soleymani Sina , Güngördü M. Zeki , Kung Patrick , Kim Seongsin M.
Published: February 2019

Infrared metasurface anomalous reflector with ultra-high efficiency and broad band operation is designed via multi-sheet graphene layer with triangular holes. The anomalous reflection angle covers the range of 10° to 90° with the efficiency higher than 80%, over a broad spectral range from 7 μm–40 μm of infrared spectrum. It reaches above 92% at the center wavelength in the spectral response. By increasing the periodicity of phase gradient, we can expand this frequency band even further without losing efficiency. The compact design of metasurface affords the adjustability of the electrochemical potential level of graphene by means of gating. Additionally, the impact of the number of graphene sheets for the optimum efficiency of the proposed structure is investigated. By adding the secondary graphene metasurface with opposite direction of phase gradient, we demonstrated the tunability of the reflection angle from θr to −θr with bias voltage.
Distinct impacts of the 1997–98 and 2015–16 extreme El Niños on Japanese eel larval catch
Authors: Lin Yong-Fu , Wu Chau-Ron
Published: February 2019

Extraordinarily poor recruitment of Japanese eels in East Asia has been generally reported during extreme El Niño years. However, the scenario failed to take place during the 2015–16 extreme event. In this study, we examined possible factors responsible for differing eel abundance in East Asia during the two strongest recent extreme El Niños, which occurred in 1997–98 and 2015–16. Numerical tracer experiments were carried out to determine why the impacts on eel catches seen in 1997–98 were not repeated in 2015–16. Among physical factors, two scenarios are likely responsible for extremely poor recruitment in East Asia: southward migration of the North Equatorial Current (NEC) or southward movement of eel spawning grounds. Comparing the latitudinal shift of NEC locations between the 1997–98 and 2015–16 El Niños, we conclude that NEC migration may be a factor, but is not chiefly responsible, for lower eel catches. Our findings pointed to southward movement of spawning grounds as the dominant factor. The northward movement of spawning grounds during 2015–16 meant that eel larvae were preferentially transported into the NEC-Kuroshio system, which resulted in a higher rate of recruitment success. The distinct evolution and dynamics of these two El Niño events led to different spawning ground locations, impacting eel abundance in East Asian countries.
The NOP-1 peptide derived from the central regulator of ethylene signaling EIN2 delays floral senescence in cut flowers
Authors: Hoppen Claudia , Müller Lena , Albrecht Anna Christina , Groth Georg
Published: February 2019

The plant hormone ethylene was identified as important triggering factor and primary regulator of flower senescence in many species. Consequently, application of chemical inhibitors of ethylene biosynthesis and action is used to extend the longevity of ethylene-sensitive flowers. Here, we show that the peptide NOP-1, a biological derived from the nuclear localization signal of ethylene regulator EIN2 tightly binds to the ethylene receptor of carnation plants - a model to study flower senescence. When applied on cut flowers the peptide biological delays petal senescence similar to previously identified and currently used chemical inhibitors, but offers significant advances to these chemicals in biodegradability, sustainability and ecotoxicity. Our bioinformatic analysis of a wide range of ethylene receptors indicates complete sequence conservation of the anticipated NOP-1 binding site in flower species supporting a widespread use of the peptide on flowering ornamentals to delay senescence and decay in cut flowers. We anticipate our innovative approach to extend flower longevity by a new class of biomolecules such as peptides, peptide analogues and peptide mimetics will significantly advance our technological capability to delay flower senescence and expand vase-life of cut flowers in a sustainable and environmentally friendly manner.
Self-assembly of dental surface nanofilaments and remineralisation by SnF2 and CPP-ACP nanocomplexes
Authors: Fernando James R. , Shen Peiyan , Sim Christina P. C. , Chen Yu-Yen , Walker Glenn D. , Yuan Yi , Reynolds Coralie , Stanton David P. , MacRae Colin M. , Reynolds Eric C.
Published: February 2019

Dental caries, erosion and hypersensitivity are major public health problems. SnF2 is used widely in oral care products to help prevent/treat these conditions. Casein phosphopeptide-stabilised amorphous calcium phosphate nanocomplexes (CPP-ACP) are a biomimetic nanotechnology of salivary phosphopeptide-ACP complexes that deliver bioavailable calcium and phosphate ions to promote dental remineralisation (repair). We show here using in vitro studies and a double-blind, randomised controlled, cross-over design in situ clinical trial that SnF2 and CPP-ACP interact to form a nanofilament coating on the tooth surface and that together they are superior in their ability to promote dental remineralisation. Sn(II) by cross-linking the CPP-ACP helps to stabilise the complexes which improves delivery to the tooth surface and enhances binding and ion incorporation into tooth mineral. The combination of SnF2 and CPP-ACP in oral care products may significantly improve their efficacy in prevention/treatment of dental caries/erosion and hypersensitivity.
Sol-gel derived boehmite nanostructures is a versatile nanoplatform for biomedical applications
Authors: Solovev Yaroslav V. , Prilepskii Artur Y. , Krivoshapkina Elena F. , Fakhardo Anna F. , Bryushkova Ekaterina A. , Kalikina Polina A. , Koshel Elena I. , Vinogradov Vladimir V.
Published: February 2019

Alumina is one of the most promising carriers for drug delivery due to the long history of its usage as a vaccine adjuvant. Sol-gel synthesis provides excellent conditions for entrapment of biomolecules within an inorganic cage providing stabilization of proteins under the extremal conditions. In this paper, we show in vitro investigation of monodisperse alumina xerogel nanocontainers (AXNCs) using bovine serum albumin as a model protein entrapped in sol-gel alumina building blocks. Particularly, dose and cell-type dependent cytotoxicity in HeLa and A549 cancer cell lines were employed as well as investigation of antibacterial effect and stability of AXNCs in different biological media. It was shown, that the release of entrapped protein could be provided only in low pH buffer (as in cancer cell cytoplasm). This property could be applied for anticancer drug development. We also discovered boehmite nanoparticles effect on horizontal gene transfer and observed the appearance of antibiotic resistance by means of exchanging of the corresponding plasmid between two different E. coli strains. The present work may help to understand better the influence of AXNCs on various biological systems, such as prokaryotic and eukaryotic cells, and the activity of AXNCs in different biological media.
Original association of ion transporters mediates the ECM-induced breast cancer cell survival: Kv10.1-Orai1-SPCA2 partnership
Authors: Peretti Marta , Badaoui Mehdi , Girault Alban , Van Gulick Laurence , Mabille Marie-Pierre , Tebbakha Riad , Sevestre Henri , Morjani Hamid , Ouadid-Ahidouch Halima
Published: February 2019

In the last years it has been shown that many components of tumor microenvironment (TM) can induce cell signaling that permit to breast cancer cells (BC) to maintain their aggressiveness. Ion channels have a role in mediating TM signal; recently we have demonstrated a functional collaboration between Kv10.1 and Orai1 channels in mediating the pro-survival effect of collagen 1 on BC cells. Here we show how SPCA2 (Secretory Pathway Ca2+ ATPase) has a role in this process and is able to support survival and proliferation induced by collagen 1. By participating to an auto-sustaining loop, SPCA2 enhances membrane expression of Kv10.1 and Orai1; the activity of every component of this trio is necessary to mediate a store independent calcium entry (SICE). This SICE is fundamental to maintain both the activation of the pro-survival pathway and the membrane localization and consequently the activity of the two channels. Moreover, the three proteins and the collagen receptor DDR1 are overexpressed only in aggressive tumors tissues. In this work, we propose a novel association between SPCA2, Kv10.1 and Orai1 involved in mediating transduction signals from TM to the BC cells that can be potentially exploited in the search of novel therapeutic targets specific to tumor tissues.
Mass Effect Deformation Heterogeneity (MEDH) on Gadolinium-contrast T1-weighted MRI is associated with decreased survival in patients with right cerebral hemisphere Glioblastoma: A feasibility study
Authors: Prasanna Prateek , Mitra Jhimli , Beig Niha , Nayate Ameya , Patel Jay , Ghose Soumya , Thawani Rajat , Partovi Sasan , Madabhushi Anant , Tiwari Pallavi
Published: February 2019

Subtle tissue deformations caused by mass-effect in Glioblastoma (GBM) are often not visually evident, and may cause neurological deficits, impacting survival. Radiomic features provide sub-visual quantitative measures to uncover disease characteristics. We present a new radiomic feature to capture mass effect-induced deformations in the brain on Gadolinium-contrast (Gd-C) T1w-MRI, and their impact on survival. Our rationale is that larger variations in deformation within functionally eloquent areas of the contralateral hemisphere are likely related to decreased survival. Displacements in the cortical and subcortical structures were measured by aligning the Gd-C T1w-MRI to a healthy atlas. The variance of deformation magnitudes was measured and defined as Mass Effect Deformation Heterogeneity (MEDH) within the brain structures. MEDH values were then correlated with overall-survival of 89 subjects on the discovery cohort, with tumors on the right (n = 41) and left (n = 48) cerebral hemispheres, and evaluated on a hold-out cohort (n = 49 subjects). On both cohorts, decreased survival time was found to be associated with increased MEDH in areas of language comprehension, social cognition, visual perception, emotion, somato-sensory, cognitive and motor-control functions, particularly in the memory areas in the left-hemisphere. Our results suggest that higher MEDH in functionally eloquent areas of the left-hemisphere due to GBM in the right-hemisphere may be associated with poor-survival.
The oncological safety in minimally invasive versus open distal pancreatectomy for pancreatic ductal adenocarcinoma: a systematic review and meta-analysis
Authors: Yang Du-Jiang , Xiong Jun-Jie , Lu Hui-Min , Wei Yi , Zhang Ling , Lu Shan , Hu Wei-Ming
Published: February 2019

The safety of minimally invasive distal pancreatectomy (MIDP) and open distal pancreatectomy (ODP) regarding oncological outcomes of pancreatic ductal adenocarcinoma (PDAC) remains inconclusive. Therefore, the aim of this study was to examine the oncological safety of MIDP and ODP for PDAC. Major databases including PubMed, Embase, Science Citation Index Expanded, and the Cochrane Library were searched for studies comparing outcomes in patients undergoing MIDP and ODP for PDAC from January 1994 to August 2018. In total, 11 retrospective comparative studies with 4829 patients (MIDP: 1076, ODP: 3753) were included. The primary outcome was long-term survival, including 3-year overall survival (OS) and 5-year OS. The 3-year OS (hazard ratio (HR): 1.03, 95% confidence interval (CI): 0.89, 1.21; P = 0.66) and 5-year OS (HR: 0.91, 95% CI: 0.65, 1.28; P = 0.59) showed no significant differences between the two groups. Furthermore, the positive surgical margin rate (weighted mean difference (WMD): 0.71, 95% CI: 0.56, 0.89, P = 0.003) was lower in the MIDP group. However, patients in the MIDP group had less intraoperative blood loss (WMD: −250.03, 95% CI: −359.68, −140.39; P < 0.00001), a shorter hospital stay (WMD: −2.76, 95% CI: −3.73, −1.78; P < 0.00001) and lower morbidity (OR: 0.57, 95% CI: 0.46, 0.71; P < 0.00001) and mortality (OR: 0.50, 95% CI: 0.31, 0.81, P = 0.005) than patients in the ODP group. The limited evidence suggested that MIDP might be safer with regard to oncological outcomes in PDAC patients. Therefore, future high-quality studies are needed to examine the oncological safety of MIDP.
Compton-based prompt gamma imaging using ordered origin ensemble algorithm with resolution recovery in proton therapy
Authors: Yao Zhiyang , Xiao Yongshun , Chen Zhiqiang , Wang Bo , Hou Qinhan
Published: February 2019

Prompt gamma ray (PG) imaging based on Compton camera (CC) is promising to realize in vivo verification during the proton therapy. However, the finite spatial and energy resolution of current CC, as well as the Doppler broaden effect, degrade the quality and resolution of PG images. In addition, due to the inherent geometrical complexity of Compton camera data, PG imaging can be time-consuming and difficult to reconstruct in real-time, while using standard techniques such as filtered back-projection or maximum likelihood-expectation maximization. In this paper, we propose three modifications of origin ensembles with resolution recovery (OE-RR) algorithm based on Markov chains to accelerate the convergence to equilibrium of OE-RR algorithm and improve the image quality. For evaluation, we performed a Monte Carlo simulation of a three-stage CZT Compton camera with resolution loss to detect the PG produced by a proton beam in a water phantom, and evaluate image quality of the gamma rays emitted during proton irradiation. The results show that our ordered OE-RR algorithm realized a good resolution recovery and accurate estimation of the position, including the peak and the distal falloff of the PG emission with remarkably faster reconstruction, thus demonstrating the feasibility of this new method in non-idealized PG-based proton range verification.
The Placental Transcriptome in Late Gestational Hypoxia Resulting in Murine Intrauterine Growth Restriction Parallels Increased Risk of Adult Cardiometabolic Disease
Authors: Chu Alison , Casero David , Thamotharan Shanthie , Wadehra Madhuri , Cosi Amy , Devaskar Sherin U.
Published: February 2019

Intrauterine growth restriction (IUGR) enhances risk for adult onset cardiovascular disease (CVD). The mechanisms underlying IUGR are poorly understood, though inadequate blood flow and oxygen/nutrient provision are considered common endpoints. Based on evidence in humans linking IUGR to adult CVD, we hypothesized that in murine pregnancy, maternal late gestational hypoxia (LG-H) exposure resulting in IUGR would result in (1) placental transcriptome changes linked to risk for later CVD, and 2) adult phenotypes of CVD in the IUGR offspring. After subjecting pregnant mice to hypoxia (10.5% oxygen) from gestational day (GD) 14.5 to 18.5, we undertook RNA sequencing from GD19 placentas. Functional analysis suggested multiple changes in structural and functional genes important for placental health and function, with maximal dysregulation involving vascular and nutrient transport pathways. Concordantly, a ~10% decrease in birthweights and ~30% decrease in litter size was observed, supportive of placental insufficiency. We also found that the LG-H IUGR offspring exhibit increased risk for CVD at 4 months of age, manifesting as hypertension, increased abdominal fat, elevated leptin and total cholesterol concentrations. In summary, this animal model of IUGR links the placental transcriptional response to the stressor of gestational hypoxia to increased risk of developing cardiometabolic disease.
Minor sequence modifications in temporin B cause drastic changes in antibacterial potency and selectivity by fundamentally altering membrane activity
Authors: Manzo Giorgia , Ferguson Philip M. , Gustilo V. Benjamin , Hind Charlotte K. , Clifford Melanie , Bui Tam T. , Drake Alex F. , Atkinson R. Andrew , Sutton J. Mark , Batoni Giovanna , Lorenz Christian D. , Phoenix David A. , Mason A. James
Published: February 2019

Antimicrobial peptides (AMPs) are a potential source of new molecules to counter the increase in antimicrobial resistant infections but a better understanding of their properties is required to understand their native function and for effective translation as therapeutics. Details of the mechanism of their interaction with the bacterial plasma membrane are desired since damage or penetration of this structure is considered essential for AMPs activity. Relatively modest modifications to AMPs primary sequence can induce substantial changes in potency and/or spectrum of activity but, hitherto, have not been predicted to substantially alter the mechanism of interaction with the bacterial plasma membrane. Here we use a combination of molecular dynamics simulations, circular dichroism, solid-state NMR and patch clamp to investigate the extent to which temporin B and its analogues can be distinguished both in vitro and in silico on the basis of their interactions with model membranes. Enhancing the hydrophobicity of the N-terminus and cationicity of the C-terminus in temporin B improves its membrane activity and potency against both Gram-negative and Gram-positive bacteria. In contrast, enhancing the cationicity of the N-terminus abrogates its ability to trigger channel conductance and renders it ineffective against Gram-positive bacteria while nevertheless enhancing its potency against Escherichia coli. Our findings suggest even closely related AMPs may target the same bacterium with fundamentally differing mechanisms of action.
Electrospun patterned porous scaffolds for the support of ovarian follicles growth: a feasibility study
Authors: Liverani Liliana , Raffel Nathalie , Fattahi Amir , Preis Alexander , Hoffmann Inge , Boccaccini Aldo R. , Beckmann Matthias W. , Dittrich Ralf
Published: February 2019

Recently, the interest of the scientific community is focused on the application of tissue engineering approach for the fertility restoration. In this paper innovative patterned electrospun fibrous scaffolds were fabricated and used as 3D system for porcine follicles culture. The obtained scaffolds demonstrated to be a suitable support which did not alter or interfere with the typical spherical follicles morphology. The fibrillar structure of the scaffolds mimics the morphology of the healthy native tissue. The use of porcine follicles implied many advantages respect to the use of mouse model. Relevant results showed that more than the scaffold pattern and struts dimension, the selection of proper biomaterials improve the follicles adhesion and development.
A continuous fluorescence assay for simple quantification of bile salt hydrolase activity in the gut microbiome
Authors: Brandvold Kristoffer R. , Weaver Jacqueline M. , Whidbey Christopher , Wright Aaron T.
Published: February 2019

The microbiota of the mammalian gut plays a dynamic role in controlling host physiology. The effect of gut microbiota activity on host health is particularly evident in the case of bile homeostasis. Bile is produced by the host and is modified by the gut microbiota, which impacts the net hydrophobicity of the total bile acid pool, and also modulates host signaling pathways. A key mechanism by which the microbiota modify bile is through deconjugation of bile salts through bile salt hydrolase (BSH) enzymatic activity, which is postulated to be a prerequisite for all further microbial metabolism. BSH activity in the gut is largely considered to be beneficial for the host, and genes encoding BSHs are found in the genomes of many taxa found in over-the-counter probiotics. Despite the therapeutic relevance of this enzyme, there is no sensitive and simple assay for continuous monitoring of BSH activity, and there are no non-destructive means of characterizing its activity in whole cell or microbial community samples. Herein, we describe a continuous fluorescence assay that can be used for characterization of BSH activity with purified protein, cell lysates, whole cells, and in human gut microbiome samples. The method is a “turn-on” reporter strategy, which employs synthetic substrates that yield a fluorescent product upon BSH-dependent turnover. This assay is used to show the first in vivo characterization of BSH activity. We also demonstrate continuous, non-destructive quantification of BSH activity in a human fecal microbiome sample containing recombinant BSH.
Determination of basal phosphodiesterase activity in mouse rod photoreceptors with cGMP clamp
Authors: Turunen Teemu T. , Koskelainen Ari
Published: February 2019

Light regulates cGMP concentration in the photoreceptor cytoplasm by activating phosphodiesterase (PDE) molecules through a G-protein signalling cascade. Spontaneous PDE activity is present in rod outer segments even in darkness. This basal PDE activity (βdark) has not been determined in wild type mammalian photoreceptor cells although it plays a key role in setting the sensitivity and recovery kinetics of rod responses. We present a novel method for determination of βdark using local electroretinography (LERG) from isolated mouse retinas. The method is based on the ability of PDE inhibitors to decrease βdark, which can be counterbalanced by increasing PDE activity with light. This procedure clamps cytoplasmic cGMP to its dark value. βdark can be calculated based on the amount of light needed for the “cGMP clamp” and information extracted from the registered rod photoresponses. Here we apply this method to determine βdark values for the first time in the mammalian rods and obtain the following estimates for different mouse models: 3.9 s−1 for wild type, 4.5 s−1 for guanylate cyclase activating proteins (GCAPs) knockout, and 4.4 s−1 for GCAPs and recoverin double knockout mice. Our results suggest that depletion of GCAPs or recoverin do not affect βdark.
Observation of Restored Topological Surface States in Magnetically Doped Topological Insulator
Authors: Kim Jinsu , Shin Eun-Ha , Sharma Manoj K. , Ihm Kyuwook , Dugerjav Otgonbayar , Hwang Chanyong , Lee Hwangho , Ko Kyung-Tae , Park Jae-Hoon , Kim Miyoung , Kim Hanchul , Jung Myung-Hwa
Published: February 2019

The introduction of ferromagnetic order in topological insulators in general breaks the time-reversal symmetry and a gap is opened in topological surface bands. Various studies have focused on gap-opened magnetic topological insulators, because such modified band structures provide a promising platform for observing exotic quantum physics. However, the role of antiferromagnetic order in topological insulators is still controversial. In this report, we demonstrate that it is possible to restore the topological surface states by effectively reducing the antiferromagnetic ordering in Gd-substituted Bi2Te3. We successfully control the magnetic impurities via thermal treatments in ultra-high vacuum condition and observe apparent restoration of topological surface band dispersions. The microscopic mechanism of atomic rearrangements and the restoration process of topological surface states are unraveled by the combination of scanning tunneling microscopy measurements and density functional theory calculations. This work provides an effective way to control the magnetic impurities which is strongly correlated with topological surface states.
Nanoparticle-mediated magnetic hyperthermia is an effective method for killing the human-infective protozoan parasite Leishmania mexicana in vitro
Authors: Berry Sarah L. , Walker Karen , Hoskins Clare , Telling Neil D. , Price Helen P.
Published: January 2019

Cutaneous leishmaniasis is a neglected tropical disease characterized by disfiguring skin lesions. Current chemotherapeutic options depend on toxic, expensive drugs that are both difficult to administer and becoming less effective due to increasing levels of resistance. In comparison, thermotherapy displays greater patient compliance and less adverse systemic effects, but there are still significant issues associated with this. The procedure is painful, requiring local anaesthetic, and is less effective against large lesions. Using nanoparticles to controllably generate heat in a localized manner may provide an alternative solution. Here we evaluate magnetic hyperthermia, using iron oxide magnetic nanoparticles, as a localized, heat-based method to kill the human-infective parasite in vitro. We assessed the effectiveness of this method against the differentiated, amastigote form of the parasite using three distinct viability assays: PrestoBlue, Live/Dead stain and a novel luciferase-based assay. Changes in amastigote morphology and ultrastructure were assessed by immunofluorescence, scanning and transmission electron microscopy. Our findings show that magnetic hyperthermia is an effective method to kill host-infective amastigotes, with morphological changes consistent with heat treatment. This method has the potential to be a step-change for research into new therapeutic options that moves away from the expensive chemotherapeutics currently dominating the research climate.
Acidogenesis, solventogenesis, metabolic stress response and life cycle changes in Clostridium beijerinckii NRRL B-598 at the transcriptomic level
Authors: Patakova Petra , Branska Barbora , Sedlar Karel , Vasylkivska Maryna , Jureckova Katerina , Kolek Jan , Koscova Pavlina , Provaznik Ivo
Published: February 2019

Clostridium beijerinckii NRRL B-598 is a sporulating, butanol and hydrogen producing strain that utilizes carbohydrates by the acetone-butanol-ethanol (ABE) fermentative pathway. The pathway consists of two metabolic phases, acidogenesis and solventogenesis, from which the latter one can be coupled with sporulation. Thorough transcriptomic profiling during a complete life cycle and both metabolic phases completed with flow cytometry, microscopy and a metabolites analysis helped to find out key genes involved in particular cellular events. The description of genes/operons that are closely involved in metabolism or the cell cycle is a necessary condition for metabolic engineering of the strain and will be valuable for all C. beijerinckii strains and other Clostridial species. The study focused on glucose transport and catabolism, hydrogen formation, metabolic stress response, binary fission, motility/chemotaxis and sporulation, which resulted in the composition of the unique image reflecting clostridial population changes. Surprisingly, the main change in expression of individual genes was coupled with the sporulation start and not with the transition from acidogenic to solventogenic metabolism. As expected, solvents formation started at pH decrease and the accumulation of butyric and acetic acids in the cultivation medium.
The Dissociable Effects of Induced Positive and Negative Moods on Cognitive Flexibility
Authors: Hsieh Shulan , Lin Siang Jyun
Published: February 2019

This study investigates how different valences of induced moods modulate cognitive flexibility in a task-switching paradigm. Forty-eight participants aged 19–25 years performed task switching after watching emotional film clips to induce an emotion (neutral, positive, or negative emotions). Two indicators of flexibility were evaluated: (1) the performance decrement reflected by increased reaction time (RT) or errors on the task-switch trial relative to a task-repetition trial, which is known as the “switching cost,” and (2) the performance improvement reflected by decreased RT or errors when switching from a task-switching context to a single-task context, which is known as the “fade-out” effect. These indicators reflect cognitive flexibility on short and long time scales, respectively. The results show that negative moods reduced switching costs, particularly in incongruent trials. In addition, negative moods were found to cause a prolonged fade-out effect compared with neutral and positive moods, indicating that participants required more trials to adjust to the single-task condition after experiencing the task-switching context. The result suggests that only negative moods and not positive moods modulated both the short and long time scales of cognitive flexibility but with dissociable effects. Possible explanations are discussed.
Identification and characterization of synthetic chondroitin-4-sulfate binding peptides in neuronal functions
Authors: Loers Gabriele , Liao Yonghong , Hu Chengliang , Xue Weikang , Shen Huifan , Zhao Weijiang , Schachner Melitta
Published: January 2019

Chondroitin sulfate proteoglycans (CSPGs), up-regulated in and around the glial scar after mammalian spinal cord injury, have been suggested to be key inhibitory molecules for functional recovery by impeding axonal regrowth/sprouting and synaptic rearrangements. CSPG-mediated inhibition is mainly associated with the glycosaminoglycan chains of CSPGs, and chondroitin-4-sulfate (C4S) is the predominant sulfated structure that regulates axonal guidance and growth in the adult nervous system. With the aim to find molecules that neutralize the inhibitory functions of C4S, we screened a phage display library for peptides binding to C4S. From the phage clones binding to C4S we selected three peptides for further analysis. We observed that these peptides bind to C4S, but not chondroitin-6-sulfate, heparin sulfate or dermatan sulfate, in a concentration-dependent and saturable manner, whereas the scrambled peptides showed highly reduced or no binding to C4S. The C4S-binding peptides, but not their scrambled counterparts, when added to cultures of mouse cerebellar neurons and human neuroblastoma cells, neutralized the inhibitory functions of the C4S- and CSPG-coated substrate on cell adhesion, neuronal migration and neurite outgrowth. These results indicate that the C4S-binding peptides neutralize several inhibitory functions of CSPGs, suggesting that they may be beneficial in repairing mammalian nervous system injuries.
Cell population balance of cardiovascular spheroids derived from human induced pluripotent stem cells
Authors: Yan Yuanwei , Bejoy Julie , Xia Junfei , Griffin Kyle , Guan Jingjiao , Li Yan
Published: February 2019

Stem cell-derived cardiomyocytes and vascular cells can be used for a variety of applications such as studying human heart development and modelling human disease in culture. In particular, protocols based on modulation of Wnt signaling were able to produce high quality of cardiomyocytes or vascular cells from human pluripotent stem cells (hPSCs). However, the mechanism behind the development of 3D cardiovascular spheroids into either vascular or cardiac cells has not been well explored. Hippo/Yes-associated protein (YAP) signaling plays important roles in the regulation of organogenesis, but its impact on cardiovascular differentiation has been less evaluated. In this study, the effects of seeding density and a change in YAP signaling on 3D cardiovascular spheroids patterning from hPSCs were evaluated. Compared to 2D culture, 3D cardiovascular spheroids exhibited higher levels of sarcomeric striations and higher length-to-width ratios of α-actinin+ cells. The spheroids with high seeding density exhibited more α-actinin+ cells and less nuclear YAP expression. The 3D cardiovascular spheroids were also treated with different small molecules, including Rho kinase inhibitor (Y27632), Cytochalasin D, Dasatinib, and Lysophosphatidic acid to modulate YAP localization. Nuclear YAP inhibition resulted in lower expression of active β-catenin, vascular marker, and MRTF, the transcription factor mediated by RhoGTPases. Y27632 also promoted the gene expression of MMP-2/-3 (matrix remodeling) and Notch-1 (Notch signaling). These results should help our understanding of the underlying effects for the efficient patterning of cardiovascular spheroids after mesoderm formation from hPSCs.
Chronic kidney disease induces left ventricular overexpression of the pro-hypertrophic microRNA-212
Authors: Sárközy Márta , Gáspár Renáta , Zvara Ágnes , Siska Andrea , Kővári Bence , Szűcs Gergő , Márványkövi Fanni , Kovács Mónika G. , Diószegi Petra , Bodai László , Zsindely Nóra , Pipicz Márton , Gömöri Kamilla , Kiss Krisztina , Bencsik Péter , Cserni Gábor , Puskás László G. , Földesi Imre , Thum Thomas , Bátkai Sándor , Csont Tamás
Published: February 2019

Chronic kidney disease (CKD) is a public health problem that increases the risk of cardiovascular morbidity and mortality. Heart failure with preserved ejection fraction (HFpEF) characterized by left ventricular hypertrophy (LVH) and diastolic dysfunction is a common cardiovascular complication of CKD. MicroRNA-212 (miR-212) has been demonstrated previously to be a crucial regulator of pathologic LVH in pressure-overload-induced heart failure via regulating the forkhead box O3 (FOXO3)/calcineurin/nuclear factor of activated T-cells (NFAT) pathway. Here we aimed to investigate whether miR-212 and its hypertrophy-associated targets including FOXO3, extracellular signal-regulated kinase 2 (ERK2), and AMP-activated protein kinase (AMPK) play a role in the development of HFpEF in CKD. CKD was induced by 5/6 nephrectomy in male Wistar rats. Echocardiography and histology revealed LVH, fibrosis, preserved systolic function, and diastolic dysfunction in the CKD group as compared to sham-operated animals eight and/or nine weeks later. Left ventricular miR-212 was significantly overexpressed in CKD. However, expressions of FOXO3, AMPK, and ERK2 failed to change significantly at the mRNA or protein level. The protein kinase B (AKT)/FOXO3 and AKT/mammalian target of rapamycin (mTOR) pathways are also proposed regulators of LVH induced by pressure-overload. Interestingly, phospho-AKT/total-AKT ratio was increased in CKD without significantly affecting phosphorylation of FOXO3 or mTOR. In summary, cardiac overexpression of miR-212 in CKD failed to affect its previously implicated hypertrophy-associated downstream targets. Thus, the molecular mechanism of the development of LVH in CKD seems to be independent of the FOXO3, ERK1/2, AMPK, and AKT/mTOR-mediated pathways indicating unique features in this form of LVH.
Epidemiological and molecular forensics of cholera recurrence in Haiti
Authors: Rebaudet Stanislas , Moore Sandra , Rossignol Emmanuel , Bogreau Hervé , Gaudart Jean , Normand Anne-Cécile , Laraque Marie-José , Adrien Paul , Boncy Jacques , Piarroux Renaud
Published: February 2019

Cholera has affected Haiti with damping waves of outbreaks since October 2010. However, mechanisms behind disease persistence during lull periods remain poorly understood. By mid 2014, cholera transmission seemed to only persist in the northern part of Haiti. Meanwhile, cholera appeared nearly extinct in the capital, Port-au-Prince, where it eventually exploded in September 2014. This study aimed to determine whether this outbreak was caused by local undetected cases or by re-importation of the disease from the north. Applying an integrated approach between November 2013 and November 2014, we assessed the temporal and spatial dynamics of cholera using routine surveillance data and performed population genetics analyses of 178 Vibrio cholerae O1 clinical isolates. The results suggest that the northern part of the country exhibited a persisting metapopulation pattern with roaming oligoclonal outbreaks that could not be effectively controlled. Conversely, undetected and unaddressed autochthonous low-grade transmission persisted in the Port-au-Prince area, which may have been the source of the acute outbreak in late-2014. Cholera genotyping is a simple but powerful tool to adapt control strategies based on epidemic specificities. In Haiti, these data have already yielded significant progress in cholera surveillance, which is a key component of the strategy to eventually eliminate cholera.