bluuebunny/crossref_metadata_2025 huggingface dataset:
Activation of miR-34a impairs autophagic flux and promotes cochlear cell death via repressing ATG9A: implications for age-related hearing loss
Authors: Pang Jiaqi , Xiong Hao , Lin Peiliang , Lai Lan , Yang Haidi , Liu Yimin , Huang Qiuhong , Chen Suijun , Ye Yongyi , Sun Yingfeng , Zheng Yiqing
Published: October 2017

Age-related hearing loss is a major unresolved public health problem. We have previously elucidated that the activation of cochlear miR-34a is correlated with age-related hearing loss in C57BL/6 mice. A growing body of evidence points that aberrant autophagy promotes cell death during the development of multiple age-related diseases. The aim of this study was to investigate the role of miR-34a-involved disorder of autophagy in the pathogenesis of age-related hearing loss. Our results showed that miR-34a expression was markedly upregulated in the aging cochlea accompanied with impairment of autophagic flux. In the inner ear HEI-OC1 cell line, miR-34a overexpression resulted in an accumulation of phagophores and impaired autophagosome–lysosome fusion, and led to cell death subsequently. Notably, autophagy-related protein 9A (ATG9A), an autophagy protein, was significantly decreased after miR-34a overexpression. Knockdown of ATG9A inhibited autophagy flux, which is similar to the effects of miR-34a overexpression. Moreover, ursodeoxycholic acid significantly rescued miR-34a-induced HEI-OC1 cell death by restoring autophagy activity. Collectively, these findings increase our understanding of the biological effects of miR-34a in the development of age-related hearing loss and highlight miR-34a as a promising therapeutic target for its treatment.
RTN1-C mediates cerebral ischemia/reperfusion injury via ER stress and mitochondria-associated apoptosis pathways
Authors: Gong Lingli , Tang Yuewen , An Ran , Lin Muya , Chen Lijian , Du Jian
Published: October 2017

The reticulon family has been found to induce apoptosis, inhibit axon regeneration and regulate protein trafficking. However, little is known about the mechanisms of how reticulon proteins are involved in neuronal death-promoting processes during ischemia. Here, we report that the expression of Reticulon Protein 1-C (RTN1-C) was associated with the progression of cerebral ischemia/reperfusion (I/R) injury. Using a combination of rat middle cerebral artery occlusion (MCAO) stroke and oxygen-glucose deprivation followed by reoxygenation (OGD/R) models, we determined that the expression of RTN1-C was significantly increased during cerebral ischemic/reperfusion. RTN1-C overexpression induced apoptosis and increased the cell vulnerability to ischemic injury, whereas RTN1-C knockdown reversed ischemia-induced apoptosis and attenuated the vulnerability of OGD/R-treated neural cells. Mechanistically, we demonstrated that RTN1-C mediated OGD/R-induced apoptosis through ER stress and mitochondria-associated pathways. RTN1-C interacted with Bcl-xL and increased its localization in the ER, thus reducing the anti-apoptotic activity of Bcl-xL. Most importantly, knockdown of Rtn1-c expression in vivo attenuated apoptosis in MCAO rats and reduced the extent of I/R-induced brain injury, as assessed by infarct volume and neurological score. Collectively, these data support for the first time that RTN1-C may represent a novel candidate for therapies against cerebral ischemia/reperfusion injury.
Reciprocal amplification of caspase-3 activity by nuclear export of a putative human RNA-modifying protein, PUS10 during TRAIL-induced apoptosis
Authors: Jana Sujata , Hsieh Andrew C , Gupta Ramesh
Published: October 2017

Pus10 is a pseudouridine synthase present in Archaea and Eukarya, but not in Bacteria and yeast. It has been suggested that the human PUS10 (DOBI) gene is needed during TRAIL-induced apoptosis. We analyzed the role of PUS10 in TRAIL-induced apoptosis by immunofluorescence, immunoblotting and several indicators of apoptosis. We examined several TRAIL-sensitive cell lines and we also examined some resistant cell lines after treatment with cycloheximide. PUS10 is mainly present in the nucleus. Early during apoptosis, PUS10 translocates to mitochondria via CRM1-mediated export with the concurrent release of cytochrome c and SMAC. Caspase-3 is required for PUS10 translocation, which reciprocally amplifies the activity of caspase-3 through the intrinsic/mitochondrial pathway. This suggests that in addition to cytoplasmic factors, nuclear factors also have a direct role in the major apoptosis pathways. However, p53 is not involved in TRAIL-induced PUS10 movement. The caspase-3-mediated movement of PUS10 and the release of mitochondrial contents enhancing caspase-3 activity creates a feedback amplification loop for caspase-3 action. Therefore, any defect in the movement or interactions of PUS10 would reduce the TRAIL sensitivity of tumor cells.
Overaccumulation of p53-mediated autophagy protects against betulinic acid-induced apoptotic cell death in colorectal cancer cells
Authors: Wang Sen , Wang Kexin , Zhang Chundong , Zhang Wanfeng , Xu Qian , Wang Yitao , Zhang Yulin , Li Yi , Zhang Ying , Zhu Huifang , Song Fangzhou , Lei Yunlong , Bu Youquan
Published: October 2017

Betulinic acid (BA) exhibits cytotoxic activity against some cancer cells. However, the molecular mechanism of BA against CRC cells was little reported. Here, we proved that BA elicited CRC cells' growth inhibition and apoptosis in a dose-dependent manner. In addition, BA treatment induced autophagy via inhibiting the AKT-MTOR signaling pathway. Inhibition of autophagy by either administration of autophagic inhibitor chloroquine or siRNA-mediated knockdown of ATG5 could augment BA-induced apoptotic cell death as well as inhibition of cell proliferation. Moreover, we found that p53 was firstly activated by short exposure to BA and then was rapidly degraded via the ubiquitin-mediated degradation pathway in both wtp53 and mutp53 CRC cells. Notably, more preferential cytotoxicity of BA was obtained in mutp53 cells (IC50 values: HT29, 125 μM; SW480, 58 μM) rather than wtp53 cells (IC50 values: HCT116, 178 μM). Further experiments demonstrated that siRNA-mediated p53 knockdown attenuated BA-induced autophagy, and forced overexpression of p53 augmented BA-induced autophagy, indicating that p53-enhanced BA-induced autophagy. Moreover, BA enhanced the sensitivity of mutp53 cells to chemotherapy drugs such as 5-FU and ADR by degradation of mutp53. Overall, our study proved that BA could induce CRC cell death by inducing apoptosis and reduce the overaccumulation of BA-induced protective autophagy by degrading wtp53 and mutp53 dependent on the ubiquitin-mediated degradation pathway to achieve killer effect, suggesting that BA might serve as a novel desirable drug for mutp53 cancer therapy.
Bypassing the Kohn-Sham equations with machine learning
Authors: Brockherde Felix , Vogt Leslie , Li Li , Tuckerman Mark E. , Burke Kieron , Müller Klaus-Robert
Published: October 2017

Last year, at least 30,000 scientific papers used the Kohn–Sham scheme of density functional theory to solve electronic structure problems in a wide variety of scientific fields. Machine learning holds the promise of learning the energy functional via examples, bypassing the need to solve the Kohn–Sham equations. This should yield substantial savings in computer time, allowing larger systems and/or longer time-scales to be tackled, but attempts to machine-learn this functional have been limited by the need to find its derivative. The present work overcomes this difficulty by directly learning the density-potential and energy-density maps for test systems and various molecules. We perform the first molecular dynamics simulation with a machine-learned density functional on malonaldehyde and are able to capture the intramolecular proton transfer process. Learning density models now allows the construction of accurate density functionals for realistic molecular systems.
Non-canonical reader modules of BAZ1A promote recovery from DNA damage
Authors: Oppikofer Mariano , Sagolla Meredith , Haley Benjamin , Zhang Hui-Min , Kummerfeld Sarah K. , Sudhamsu Jawahar , Flynn E. Megan , Bai Tianyi , Zhang Jennifer , Ciferri Claudio , Cochran Andrea G.
Published: October 2017

Members of the ISWI family of chromatin remodelers mobilize nucleosomes to control DNA accessibility and, in some cases, are required for recovery from DNA damage. However, it remains poorly understood how the non-catalytic ISWI subunits BAZ1A and BAZ1B might contact chromatin to direct the ATPase SMARCA5. Here, we find that the plant homeodomain of BAZ1A, but not that of BAZ1B, has the unusual function of binding DNA. Furthermore, the BAZ1A bromodomain has a non-canonical gatekeeper residue and binds relatively weakly to acetylated histone peptides. Using CRISPR-Cas9-mediated genome editing we find that BAZ1A and BAZ1B each recruit SMARCA5 to sites of damaged chromatin and promote survival. Genetic engineering of structure-designed bromodomain and plant homeodomain mutants reveals that reader modules of BAZ1A and BAZ1B, even when non-standard, are critical for DNA damage recovery in part by regulating ISWI factors loading at DNA lesions and supporting transcriptional programs required for survival.
High-fidelity DNA replication in Mycobacterium tuberculosis relies on a trinuclear zinc center
Authors: Baños-Mateos Soledad , van Roon Anne-Marie M. , Lang Ulla F. , Maslen Sarah L. , Skehel J. Mark , Lamers Meindert H.
Published: October 2017

High-fidelity DNA replication depends on a proofreading 3′–5′ exonuclease that is associated with the replicative DNA polymerase. The replicative DNA polymerase DnaE1 from the major pathogen Mycobacterium tuberculosis (Mtb) uses its intrinsic PHP-exonuclease that is distinct from the canonical DEDD exonucleases found in the Escherichia coli and eukaryotic replisomes. The mechanism of the PHP-exonuclease is not known. Here, we present the crystal structure of the Mtb DnaE1 polymerase. The PHP-exonuclease has a trinuclear zinc center, coordinated by nine conserved residues. Cryo-EM analysis reveals the entry path of the primer strand in the PHP-exonuclease active site. Furthermore, the PHP-exonuclease shows a striking similarity to E. coli endonuclease IV, which provides clues regarding the mechanism of action. Altogether, this work provides important insights into the PHP-exonuclease and reveals unique properties that make it an attractive target for novel anti-mycobacterial drugs.
The metabolic enzyme fructose-1,6-bisphosphate aldolase acts as a transcriptional regulator in pathogenic Francisella
Authors: Ziveri Jason , Tros Fabiola , Guerrera Ida Chiara , Chhuon Cerina , Audry Mathilde , Dupuis Marion , Barel Monique , Korniotis Sarantis , Fillatreau Simon , Gales Lara , Cahoreau Edern , Charbit Alain
Published: October 2017

The enzyme fructose-bisphosphate aldolase occupies a central position in glycolysis and gluconeogenesis pathways. Beyond its housekeeping role in metabolism, fructose-bisphosphate aldolase has been involved in additional functions and is considered as a potential target for drug development against pathogenic bacteria. Here, we address the role of fructose-bisphosphate aldolase in the bacterial pathogen Francisella novicida. We demonstrate that fructose-bisphosphate aldolase is important for bacterial multiplication in macrophages in the presence of gluconeogenic substrates. In addition, we unravel a direct role of this metabolic enzyme in transcription regulation of genes katG and rpoA, encoding catalase and an RNA polymerase subunit, respectively. We propose a model in which fructose-bisphosphate aldolase participates in the control of host redox homeostasis and the inflammatory immune response.
Quorum sensing integrates environmental cues, cell density and cell history to control bacterial competence
Authors: Moreno-Gámez Stefany , Sorg Robin A. , Domenech Arnau , Kjos Morten , Weissing Franz J. , van Doorn G. Sander , Veening Jan-Willem
Published: October 2017

Streptococcus pneumoniae becomes competent for genetic transformation when exposed to an autoinducer peptide known as competence-stimulating peptide (CSP). This peptide was originally described as a quorum-sensing signal, enabling individual cells to regulate competence in response to population density. However, recent studies suggest that CSP may instead serve as a probe for sensing environmental cues, such as antibiotic stress or environmental diffusion. Here, we show that competence induction can be simultaneously influenced by cell density, external pH, antibiotic-induced stress, and cell history. Our experimental data is explained by a mathematical model where the environment and cell history modify the rate at which cells produce or sense CSP. Taken together, model and experiments indicate that autoinducer concentration can function as an indicator of cell density across environmental conditions, while also incorporating information on environmental factors or cell history, allowing cells to integrate cues such as antibiotic stress into their quorum-sensing response. This unifying perspective may apply to other debated quorum-sensing systems.
Molecular identification of the wheat male fertility gene Ms1 and its prospects for hybrid breeding
Authors: Tucker Elise J. , Baumann Ute , Kouidri Allan , Suchecki Radoslaw , Baes Mathieu , Garcia Melissa , Okada Takashi , Dong Chongmei , Wu Yongzhong , Sandhu Ajay , Singh Manjit , Langridge Peter , Wolters Petra , Albertsen Marc C. , Cigan A. Mark , Whitford Ryan
Published: October 2017

The current rate of yield gain in crops is insufficient to meet the predicted demands. Capturing the yield boost from heterosis is one of the few technologies that offers rapid gain. Hybrids are widely used for cereals, maize and rice, but it has been a challenge to develop a viable hybrid system for bread wheat due to the wheat genome complexity, which is both large and hexaploid. Wheat is our most widely grown crop providing 20% of the calories for humans. Here, we describe the identification of Ms1, a gene proposed for use in large-scale, low-cost production of male-sterile (ms) female lines necessary for hybrid wheat seed production. We show that Ms1 completely restores fertility to ms1d, and encodes a glycosylphosphatidylinositol-anchored lipid transfer protein, necessary for pollen exine development. This represents a key step towards developing a robust hybridization platform in wheat.
An NF-κB-microRNA regulatory network tunes macrophage inflammatory responses
Authors: Mann Mati , Mehta Arnav , Zhao Jimmy L. , Lee Kevin , Marinov Georgi K. , Garcia-Flores Yvette , Lu Li-Fan , Rudensky Alexander Y. , Baltimore David
Published: October 2017

The innate inflammatory response must be tightly regulated to ensure effective immune protection. NF-κB is a key mediator of the inflammatory response, and its dysregulation has been associated with immune-related malignancies. Here, we describe a miRNA-based regulatory network that enables precise NF-κB activity in mouse macrophages. Elevated miR-155 expression potentiates NF-κB activity in miR-146a-deficient mice, leading to both an overactive acute inflammatory response and chronic inflammation. Enforced miR-155 expression overrides miR-146a-mediated repression of NF-κB activation, thus emphasizing the dominant function of miR-155 in promoting inflammation. Moreover, miR-155-deficient macrophages exhibit a suboptimal inflammatory response when exposed to low levels of inflammatory stimuli. Importantly, we demonstrate a temporal asymmetry between miR-155 and miR-146a expression during macrophage activation, which creates a combined positive and negative feedback network controlling NF-κB activity. This miRNA-based regulatory network enables a robust yet time-limited inflammatory response essential for functional immunity.
Electrical magnetochiral effect induced by chiral spin fluctuations
Authors: Yokouchi T. , Kanazawa N. , Kikkawa A. , Morikawa D. , Shibata K. , Arima T. , Taguchi Y. , Kagawa F. , Tokura Y.
Published: October 2017

Chirality of matter can produce unique responses in optics, electricity and magnetism. In particular, magnetic crystals transmit their handedness to the magnetism via antisymmetric exchange interaction of relativistic origin, producing helical spin orders as well as their fluctuations. Here we report for a chiral magnet MnSi that chiral spin fluctuations manifest themselves in the electrical magnetochiral effect, i.e. the nonreciprocal and nonlinear response characterized by the electrical resistance depending on inner product of current and magnetic field. Prominent electrical magnetochiral signals emerge at specific temperature-magnetic field-pressure regions: in the paramagnetic phase just above the helical ordering temperature and in the partially-ordered topological spin state at low temperatures and high pressures, where thermal and quantum spin fluctuations are conspicuous in proximity of classical and quantum phase transitions, respectively. The finding of the asymmetric electron scattering by chiral spin fluctuations may explore new electromagnetic functionality in chiral magnets.
Replication fork reversal triggers fork degradation in BRCA2-defective cells
Authors: Mijic Sofija , Zellweger Ralph , Chappidi Nagaraja , Berti Matteo , Jacobs Kurt , Mutreja Karun , Ursich Sebastian , Ray Chaudhuri Arnab , Nussenzweig Andre , Janscak Pavel , Lopes Massimo
Published: October 2017

Besides its role in homologous recombination, the tumor suppressor BRCA2 protects stalled replication forks from nucleolytic degradation. Defective fork stability contributes to chemotherapeutic sensitivity of BRCA2-defective tumors by yet-elusive mechanisms. Using DNA fiber spreading and direct visualization of replication intermediates, we report that reversed replication forks are entry points for fork degradation in BRCA2-defective cells. Besides MRE11 and PTIP, we show that RAD52 promotes stalled fork degradation and chromosomal breakage in BRCA2-defective cells. Inactivation of these factors restores reversed fork frequency and chromosome integrity in BRCA2-defective cells. Conversely, impairing fork reversal prevents fork degradation, but increases chromosomal breakage, uncoupling fork protection, and chromosome stability. We propose that BRCA2 is dispensable for RAD51-mediated fork reversal, but assembles stable RAD51 nucleofilaments on regressed arms, to protect them from degradation. Our data uncover the physiopathological relevance of fork reversal and illuminate a complex interplay of homologous recombination factors in fork remodeling and stability.
The recording and characteristics of pulmonary rehabilitation in patients with COPD using The Health Information Network (THIN) primary care database
Authors: Hakamy Ali , McKeever Tricia M. , Gibson Jack E. , Bolton Charlotte E.
Published: October 2017

Pulmonary rehabilitation is recommended for patients with COPD to improve physical function, breathlessness and quality of life. Using The Health Information Network (THIN) primary care database in UK, we compared the demographic and clinical parameters of patients with COPD in relation to coding of pulmonary rehabilitation, and to investigate whether there is a survival benefit from pulmonary rehabilitation. We identified patients with COPD, diagnosed from 2004 and extracted information on demographics, pulmonary rehabilitation and clinical parameters using the relevant Read codes. Thirty six thousand one hundred and eighty nine patients diagnosed with COPD were included with a mean (SD) age of 67 (11) years, 53% were male and only 9.8% had a code related to either being assessed, referred, or completing pulmonary rehabilitation ever. Younger age at diagnosis, better socioeconomic status, worse dyspnoea score, current smoking, and higher comorbidities level are more likely to have a record of pulmonary rehabilitation. Of those with a recorded MRC of 3 or worse, only 2057 (21%) had a code of pulmonary rehabilitation. Survival analysis revealed that patients with coding for pulmonary rehabilitation were 22% (95% CI 0.69–0.88) less likely to die than those who had no coding. In UK THIN records, a substantial proportion of eligible patients with COPD have not had a coded pulmonary rehabilitation record. Survival was improved in those with PR record but coding for other COPD treatments were also better in this group. GP practices need to improve the coding for PR to highlight any unmet need locally.
Photonic Sorting of Aligned, Crystalline Carbon Nanotube Textiles
Authors: Bulmer John S. , Gspann Thurid S. , Orozco Francisco , Sparkes Martin , Koerner Hilmar , Di Bernardo A. , Niemiec Arkadiusz , Robinson J. W. A. , Koziol Krzysztof K. , Elliott James A. , O’Neill William
Published: October 2017

Floating catalyst chemical vapor deposition uniquely generates aligned carbon nanotube (CNT) textiles with individual CNT lengths magnitudes longer than competing processes, though hindered by impurities and intrinsic/extrinsic defects. We present a photonic-based post-process, particularly suited for these textiles, that selectively removes defective CNTs and other carbons not forming a threshold thermal pathway. In this method, a large diameter laser beam rasters across the surface of a partly aligned CNT textile in air, suspended from its ends. This results in brilliant, localized oxidation, where remaining material is an optically transparent film comprised of few-walled CNTs with profound and unique improvement in microstructure alignment and crystallinity. Raman spectroscopy shows substantial D peak suppression while preserving radial breathing modes. This increases the undoped, specific electrical conductivity at least an order of magnitude to beyond that of single-crystal graphite. Cryogenic conductivity measurements indicate intrinsic transport enhancement, opposed to simply removing nonconductive carbons/residual catalyst.
Digital nonlinearity compensation in high-capacity optical communication systems considering signal spectral broadening effect
Authors: Xu Tianhua , Karanov Boris , Shevchenko Nikita A. , Lavery Domaniç , Liga Gabriele , Killey Robert I. , Bayvel Polina
Published: October 2017

Nyquist-spaced transmission and digital signal processing have proved effective in maximising the spectral efficiency and reach of optical communication systems. In these systems, Kerr nonlinearity determines the performance limits, and leads to spectral broadening of the signals propagating in the fibre. Although digital nonlinearity compensation was validated to be promising for mitigating Kerr nonlinearities, the impact of spectral broadening on nonlinearity compensation has never been quantified. In this paper, the performance of multi-channel digital back-propagation (MC-DBP) for compensating fibre nonlinearities in Nyquist-spaced optical communication systems is investigated, when the effect of signal spectral broadening is considered. It is found that accounting for the spectral broadening effect is crucial for achieving the best performance of DBP in both single-channel and multi-channel communication systems, independent of modulation formats used. For multi-channel systems, the degradation of DBP performance due to neglecting the spectral broadening effect in the compensation is more significant for outer channels. Our work also quantified the minimum bandwidths of optical receivers and signal processing devices to ensure the optimal compensation of deterministic nonlinear distortions.
Linked magnolol dimer as a selective PPARγ agonist – Structure-based rational design, synthesis, and bioactivity evaluation
Authors: Dreier Dominik , Latkolik Simone , Rycek Lukas , Schnürch Michael , Dymáková Andrea , Atanasov Atanas G. , Ladurner Angela , Heiss Elke H. , Stuppner Hermann , Schuster Daniela , Mihovilovic Marko D. , Dirsch Verena M.
Published: October 2017

The nuclear receptors peroxisome proliferator-activated receptor γ (PPARγ) and its hetero-dimerization partner retinoid X receptor α (RXRα) are considered as drug targets in the treatment of diseases like the metabolic syndrome and diabetes mellitus type 2. Effort has been made to develop new agonists for PPARγ to obtain ligands with more favorable properties than currently used drugs. Magnolol was previously described as dual agonist of PPARγ and RXRα. Here we show the structure-based rational design of a linked magnolol dimer within the ligand binding domain of PPARγ and its synthesis. Furthermore, we evaluated its binding properties and functionality as a PPARγ agonist in vitro with the purified PPARγ ligand binding domain (LBD) and in a cell-based nuclear receptor transactivation model in HEK293 cells. We determined the synthesized magnolol dimer to bind with much higher affinity to the purified PPARγ ligand binding domain than magnolol (Ki values of 5.03 and 64.42 nM, respectively). Regarding their potency to transactivate a PPARγ-dependent luciferase gene both compounds were equally effective. This is likely due to the PPARγ specificity of the newly designed magnolol dimer and lack of RXRα-driven transactivation activity by this dimeric compound.
Glomalin contributed more to carbon, nutrients in deeper soils, and differently associated with climates and soil properties in vertical profiles
Authors: Wang Wenjie , Zhong Zhaoliang , Wang Qiong , Wang Humei , Fu Yujie , He Xingyuan
Published: October 2017

Despite vital importance in soil conditioning and a proxy for arbuscular mycorrhizal (AMF), glomalin-related soil protein (GRSP) contribution to soil carbon and nutrients at vertical soil profiles and underlying mechanism were not well-defined yet. Thus, 360 soil samples were collected from 72 farmland 1-m soil profiles in northeastern China, and soil physiochemical properties, nutrients, glomalin characteristics, local climates were determined. Linear decreases of glomalin amounts were observed from the top to deep soils, and glomalin/SOC (glomalin ratio to total SOC) in the 80–100 cm soil (EEG, easily-extracted GRSP, 2.2%; TG, total GRSP, 19%) was 1.34–1.5-fold higher than did in the 0–20 cm soil. Different statistical analyses crosschecked that the lower pH and higher SOC usually accompanied with the higher EEG and TG, while EEG was more sensitive to climates; Moreover, glomalin was more physiochemical-regulated in the deep soils, but more nutrient-regulation was found in the surface soils. Structure Equation Model showed that soil depths and climates indirectly affected TG and EEG features through soil properties, except significant direct effects on EEG. In future, glomalin assessment should fully consider these for identifying the AMF importance in the whole 1-m profile, and our findings also favor degrade soil improvement from glomalin rehabilitation.
The thiazide sensitive sodium chloride co-transporter NCC is modulated by site-specific ubiquitylation
Authors: Rosenbaek Lena L. , Rizzo Federica , Wu Qi , Rojas-Vega Lorena , Gamba Gerardo , MacAulay Nanna , Staub Olivier , Fenton Robert A.
Published: October 2017

The renal sodium chloride cotransporter, NCC, in the distal convoluted tubule is important for maintaining body Na+ and K+ homeostasis. Endogenous NCC is highly ubiquitylated, but the role of individual ubiquitylation sites is not established. Here, we assessed the role of 10 ubiquitylation sites for NCC function. Transient transfections of HEK293 cells with human wildtype (WT) NCC or various K to R mutants identified greater membrane abundance for K706R, K828R and K909R mutants. Relative to WT-NCC, stable tetracycline inducible MDCKI cell lines expressing K706R, K828R and K909R mutants had significantly higher total and phosphorylated NCC levels at the apical plasma membrane under basal conditions. Low chloride stimulation increased membrane abundance of all mutants to similar or greater levels than WT-NCC. Under basal conditions K828R and K909R mutants had less ubiquitylated NCC in the plasma membrane, and all mutants displayed reduced NCC ubiquitylation following low chloride stimulation. Thiazide-sensitive sodium-22 uptakes were elevated in the mutants and internalization from the plasma membrane was significantly less than WT-NCC. K909R had increased half-life, whereas chloroquine or MG132 treatment indicated that K706 and K909 play roles in lysosomal and proteasomal NCC degradation, respectively. In conclusion, site-specific ubiquitylation of NCC plays alternative roles for NCC function.
Development of a Cell-Based Assay for Measuring Base Excision Repair Responses
Authors: Golato Tyler , Brenerman Boris , McNeill Daniel R. , Li Jianfeng , Sobol Robert W. , Wilson David M.
Published: October 2017

Base excision repair (BER) is the predominant pathway for coping with most forms of hydrolytic, oxidative or alkylative DNA damage. Measuring BER capacity in living cells is valuable for both basic science applications and epidemiological studies, since deficiencies in this pathway have been associated with cancer susceptibility and other adverse health outcomes. At present, there is an ongoing effort to develop methods to effectively quantify the rate of BER as a whole. We present a variation of a previously described “Oligonucleotide Retrieval Assay” designed to measure DNA excision repair that is capable of quantifying the rate of repair of thymine glycol in a variety of human cells with a high degree of sensitivity.
Danshenol A inhibits TNF-α-induced expression of intercellular adhesion molecule-1 (ICAM-1) mediated by NOX4 in endothelial cells
Authors: Zhao Wenwen , Feng Haitao , Guo Shuhui , Han Yantao , Chen Xiuping
Published: October 2017

ICAM-1 overexpression and subsequent adhesion of leukocytes to endothelial cells play critical roles in the early stage of atherosclerosis. Danshenol A (DA) is an abietane-type diterpenoid isolated from traditional Chinese herb Salvia miltiorrhiza Bunge. The mechanisms under its regulation of adhesion of molecular expression are explored. Here, the effect of DA on TNF-α-induced ICAM-1 expression was investigated in endothelial cells. TNF-α-induced ICAM-1 expression and subsequent adhesion of monocytes, as well as elevated reactive oxygen species (ROS) generation and NOX4 expression were all significantly reversed by DA, siNOX4 and NOX4 inhibitor GKT137831. Furthermore, TNF-α-induced ICAM-1 expression, which was increased via IKKβ/IκBα-mediated activation of NF-κB p65, was also inhibited by DA. Interestingly, NOX4 overexpression suppressed the ICAM-1 expression, and this finding may be ascribed to the activation of Nrf-2. Additionally, NF-κB inhibitor PDTC, siNOX4, or DA can decrease the TNF-α-induced ICAM-1 expression and suppress the adhesion of monocytes. In all, DA inhibited TNF-α-induced ICAM-1 expression and subsequent monocyte adhesion to endothelial cells through the NOX4-dependent IKKβ/NF-κB pathway. Besides, NOX4 played dual role in regulating ICAM-1 expression via diverse signal pathway. This novel bioactivity will make DA a good candidate to be further explored for therapeutic or preventive application for atherosclerosis.
Anodal tDCS applied during multitasking training leads to transferable performance gains
Authors: Filmer Hannah L. , Lyons Maxwell , Mattingley Jason B. , Dux Paul E.
Published: October 2017

Cognitive training can lead to performance improvements that are specific to the tasks trained. Recent research has suggested that transcranial direct current stimulation (tDCS) applied during training of a simple response-selection paradigm can broaden performance benefits to an untrained task. Here we assessed the impact of combined tDCS and training on multitasking, stimulus-response mapping specificity, response-inhibition, and spatial attention performance in a cohort of healthy adults. Participants trained over four days with concurrent tDCS – anodal, cathodal, or sham – applied to the left prefrontal cortex. Immediately prior to, 1 day after, and 2 weeks after training, performance was assessed on the trained multitasking paradigm, an untrained multitasking paradigm, a go/no-go inhibition task, and a visual search task. Training combined with anodal tDCS, compared with training plus cathodal or sham stimulation, enhanced performance for the untrained multitasking paradigm and visual search tasks. By contrast, there were no training benefits for the go/no-go task. Our findings demonstrate that anodal tDCS combined with multitasking training can extend to untrained multitasking paradigms as well as spatial attention, but with no extension to the domain of response inhibition.
Novel mechanism of regulation of the 5-lipoxygenase/leukotriene B4 pathway by high-density lipoprotein in macrophages
Authors: Tsuda Shigeyasu , Shinohara Masakazu , Oshita Toshihiko , Nagao Manabu , Tanaka Nobuaki , Mori Takeshige , Hara Tetsuya , Irino Yasuhiro , Toh Ryuji , Ishida Tatsuro , Hirata Ken-ichi
Published: October 2017

High-density lipoprotein (HDL) interacts with various cells, particularly macrophages, in functional cell-HDL interactions. Here, we found that HDL protein quality and lipid quality play critical roles in HDL functions. HDL fractions from healthy volunteers (HDLHealthy) and patients with recurrent coronary atherosclerotic disease (HDLCAD) were prepared. To analyse functional HDL-macrophage interactions, macrophages were co-incubated with each HDL, and lipid mediator production was assessed by liquid chromatography/mass spectrometry-based metabololipidomics. HDLHealthy treatment attenuated the pro-inflammatory lipid mediator production, particularly that of leukotriene (LT) B4, and this treatment enhanced lipoxin (LX) B4 and resolvin (Rv) E2 production. HDLHealthy treatment enhanced the proteasome-mediated degradation of the LTB4-producing enzyme 5-lipoxygenase (LO) in activated macrophages; however, HDLCAD did not show these anti-inflammatory effects. HDLHealthy was engulfed by macrophages via clathrin-mediated endocytosis, which was a critical step in 5-LO/LTB4 regulation. We also found that HDLCAD showed higher levels of the LTB4-producing enzymes and thus promoted LTB4 production from HDLCAD. In addition, LTB4 attenuated HDL endocytosis, HDL-mediated 5-LO degradation in macrophages, and HDL-derived augmentation of macrophage phagocytosis. These results indicated that local LTB4 produced de novo from HDLCAD regulates HDL-macrophage functional interactions and plays critical roles in dysfunctional, inflammatory HDL characteristics.
Imaging mass spectrometry analysis of ubiquinol localization in the mouse brain following short-term administration
Authors: Tatsuta Yukina , Kasai Kazuaki , Maruyama Chitose , Hamano Yoshimitsu , Matsuo Kazuhiko , Katano Hajime , Taira Shu
Published: October 2017

We analyzed the localization of ubiquinol, the reduced form of coenzyme Q10 (Re-CoQ10), in mouse brain sections using matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance imaging mass spectrometry (IMS) to evaluate the effect of dietary Re-CoQ10 in mouse brain. Mice were orally administered Re-CoQ10 for 14 days and brain Re-CoQ10 content was subsequently quantified using liquid chromatography-mass spectrometry. IMS was employed to visualize Re-CoQ10 at a resolution of 150 μm in the mouse brain. Increased Re-CoQ10 was observed in the corpus callosum, hippocampus, midbrain, cerebellum, brain stem, substantia nigra and striatum. These regions are related to movement, memory and vital life functions. Thus, we demonstrated the effect of Re-CoQ10 administration on the specific localization of Re-CoQ10 in the brain.
Interleukin 17A as a good predictor of the severity of Mycoplasma pneumoniae pneumonia in children
Authors: Yang Mingyue , Meng Fanzheng , Wang Kuo , Gao Man , Lu Ruihua , Li Mengyao , Zhao Fangxing , Huang Lijuan , Zhang Yining , Cheng Genhong , Wang Xiaosong
Published: October 2017

Early distinction between severe Mycoplasma pneumoniae pneumonia (MPP) and mild MPP is still difficult. The aim of this study was to analyze cytokines in bronchoalveolar lavage fluid (BALF) and explore predicting factors of severe MPP in children. Retrospective analysis was performed on 150 children with MPP or bronchial foreign body (FB) admitted in our hospital. The mRNA levels of IL17A were found significantly lower in severe MPP group comparing with mild MPP group or FB group. However, no significant difference was found in the levels of IL4, IL10 or interferon beta1 (IFNβ1) between the two groups. Receiver operator characteristic (ROC) curve analysis showed that IL17A can be used to distinguish severe MPP from mild MPP. These results were confirmed in a validation cohort including 40 MPP children from another hospital. IL17A levels were correlated with some clinical characters, such as refractoriness and pleural effusion. Lower IL17A levels were more likely to be found in refractory MPP children or in MPP children with pleural effusion. Moreover, the protein levels of IL17A in BALF were also found greatly decreased in children with severe MPP. Thus, decreased IL17A levels in BALF may be a valuable biomarker to identify severe MPP in children.
Efficient Single-Photon Coupling from a Nitrogen-Vacancy Center Embedded in a Diamond Nanowire Utilizing an Optical Nanofiber
Authors: Yonezu Yuya , Wakui Kentaro , Furusawa Kentaro , Takeoka Masahiro , Semba Kouichi , Aoki Takao
Published: October 2017

Nitrogen-Vacancy (NV) centers in diamond are promising solid-state quantum emitters that can be utilized for photonic quantum applications. Various diamond nanophotonic devices have been fabricated for efficient extraction of single photons emitted from NV centers to a single guided mode. However, for constructing scalable quantum networks, further efficient coupling of single photons to a guided mode of a single-mode fiber (SMF) is indispensable and a difficult challenge. Here, we propose a novel efficient hybrid system between an optical nanofiber and a cylindrical-structured diamond nanowire. The maximum coupling efficiency as high as 75% for the sum of both fiber ends is obtained by numerical simulations. The proposed hybrid system will provide a simple and efficient interface between solid-state quantum emitters and a SMF suitable for constructing scalable quantum networks.
Human TAUP301L overexpression results in TAU hyperphosphorylation without neurofibrillary tangles in adult zebrafish brain
Authors: Cosacak Mehmet I. , Bhattarai Prabesh , Bocova Ledio , Dzewas Tim , Mashkaryan Violeta , Papadimitriou Christos , Brandt Kerstin , Hollak Heike , Antos Christopher L. , Kizil Caghan
Published: October 2017

Microtubule-associated TAU protein is a pathological hallmark in Alzheimer’s disease (AD), where hyperphosphorylation of TAU generates neurofibrillary tangles. To investigate the effects of TAU in a regenerative adult vertebrate brain system, we generated a cre/lox-based transgenic model of zebrafish that chronically expresses human TAUP301L, which is a variant of human TAU protein that forms neurofibrillary tangles in mouse models and humans. Interestingly, we found that although chronic and abundant expression of TAUP301L starting from early embryonic development led to hyperphosphorylation, TAUP301L did not form oligomers and neurofibrillary tangles, and did not cause elevated apoptosis and microglial activation, which are classical symptoms of tauopathies in mammals. Additionally, TAUP301L neither increased neural stem cell proliferation nor activated the expression of regenerative factor Interleukin-4, indicating that TAUP301L toxicity is prevented in the adult zebrafish brain. By combining TAUP301L expression with our established Aβ42 toxicity model, we found that Aβ42 ceases to initiate neurofibrillary tangle formation by TAUP301L, and TAUP301L does not exacerbate the toxicity of Aβ42. Therefore, our results propose a cellular mechanism that protects the adult zebrafish brain against tauopathies, and our model can be used to understand how TAU toxicity can be prevented in humans.
A turn-on endoplasmic reticulum-targeted two-photon fluorescent probe for hydrogen sulfide and bio-imaging applications in living cells, tissues, and zebrafish
Authors: Tang Yonghe , Xu An , Ma Yanyan , Xu Gaoping , Gao Shiying , Lin Weiying
Published: October 2017

As one of the important gas signal molecules, hydrogen sulfide (H2S) is associated with many important physiological processes in living organisms. Organelles, especially endoplasmic reticulum (ER), play a crucial role in the cell metabolism. Accordingly, the detection of H2S in the ER is of high interest. Toward this goal, we have described the development of the first ER-targeted fluorescent H2S probe (Na-H
2
S-ER). The new probe exhibited favorable features, such as a large turn-on fluorescence signal (45-fold fluorescence enhancement), high sensitivity and selectivity. The probe was successfully employed for imaging exogenous and endogenous H2S in the living HeLa cells. Significantly, the new probe Na-H
2
S-ER was employed to visualize H2S in the ER of living cells for the first time. In addition, the probe was also successfully used for imaging H2S in the living tissues up to a depth of 100 μm and in the living zebrafish.
Correlation of X-ray diffraction signatures of breast tissue and their histopathological classification
Authors: Moss Robert M. , Amin Amany S. , Crews Chiaki , Purdie Colin A. , Jordan Lee B. , Iacoviello Francesco , Evans Andrew , Speller Robert D. , Vinnicombe Sarah J.
Published: October 2017

This pilot study examines the correlation of X-ray diffraction (XRD) measurements with the histopathological analysis of breast tissue. Eight breast cancer samples were investigated. Each sample contained a mixture of normal and cancerous tissues. In total, 522 separate XRD measurements were made at different locations across the samples (8 in total). The resulting XRD spectra were subjected to principal component analysis (PCA) in order to determine if there were any distinguishing features that could be used to identify different tissue components. 99.0% of the variation between the spectra were described by the first two principal components (PC). Comparing the location of points in PC space with the classification determined by histopathology indicated correlation between the shape/magnitude of the XRD spectra and the tissue type. These results are encouraging and suggest that XRD could be used for the intraoperative or postoperative classification of bulk tissue samples.
Deficiency of peroxiredoxin 6 or inhibition of its phospholipase A2 activity impair the in vitro sperm fertilizing competence in mice
Authors: Moawad Adel R. , Fernandez Maria C. , Scarlata Eleonora , Dodia Chandra , Feinstein Sheldon I. , Fisher Aron B. , O’Flaherty Cristian
Published: October 2017

Prdx6−/− male mice are subfertile, and the deficiency or inactivation of Peroxiredoxins (PRDXs) is associated with human male infertility. We elucidate the impact of the lack of PRDX6 or inhibition of its calcium-independent phospholipase A2 (Ca2+-iPLA2) activity by MJ33 on fertilization competence of mouse spermatozoa. Sperm motility, viability, fertilization and blastocyst rates were lower in Prdx6−/− spermatozoa than in C57BL/6J wild-type (WT) controls (p ≤ 0.05). MJ33 inhibited the PRDX6 Ca2+-iPLA2 activity and reduced these parameters in WT spermatozoa compared with controls (p ≤ 0.05). Levels of lipid peroxidation and of superoxide anion (O2•─) were higher in Prdx6−/− than in WT spermatozoa (p ≤ 0.05). MJ33 increased the levels of lipid peroxidation and mitochondrial O2•─ production in treated versus non-treated WT spermatozoa. Acrosome reaction, binding to zona pellucida and fusion with the oolemma were lower in Prdx6−/− capacitated spermatozoa than WT capacitated controls and lower in WT spermatozoa treated with the PRDX6 inhibitor. In conclusion, the inhibition of the PRDX6 Ca2+-iPLA2 activity promotes an oxidative stress affecting viability, motility, and the ability of mouse spermatozoa to fertilize oocytes. Thus, PRDX6 has a critical role in the protection of the mouse spermatozoon against oxidative stress to assure fertilizing competence.
Low immunogenicity of mouse induced pluripotent stem cell-derived neural stem/progenitor cells
Authors: Itakura Go , Ozaki Masahiro , Nagoshi Narihito , Kawabata Soya , Nishiyama Yuichiro , Sugai Keiko , Iida Tsuyoshi , Kashiwagi Rei , Ookubo Toshiki , Yastake Kaori , Matsubayashi Kohei , Kohyama Jun , Iwanami Akio , Matsumoto Morio , Nakamura Masaya , Okano Hideyuki
Published: October 2017

Resolving the immunogenicity of cells derived from induced pluripotent stem cells (iPSCs) remains an important challenge for cell transplant strategies that use banked allogeneic cells. Thus, we evaluated the immunogenicity of mouse fetal neural stem/progenitor cells (fetus-NSPCs) and iPSC-derived neural stem/progenitor cells (iPSC-NSPCs) both in vitro and in vivo. Flow cytometry revealed the low expression of immunological surface antigens, and these cells survived in all mice when transplanted syngeneically into subcutaneous tissue and the spinal cord. In contrast, an allogeneic transplantation into subcutaneous tissue was rejected in all mice, and allogeneic cells transplanted into intact and injured spinal cords survived for 3 months in approximately 20% of mice. In addition, cell survival was increased after co-treatment with an immunosuppressive agent. Thus, the immunogenicity and post-transplantation immunological dynamics of iPSC-NSPCs resemble those of fetus-NSPCs.
Higher Expression of Toll-Like Receptors 2 and 4 in Blood Cells of Keratoconus Patiens
Authors: Sobrino Tomás , Regueiro Uxía , Malfeito Mercedes , Vieites-Prado Alba , Pérez-Mato María , Campos Francisco , Lema Isabel
Published: October 2017

Inflammation may play a significant role in Keratoconus (KC), but the implication of immunity on this inflammatory response is unknown. Therefore, our aim was to determine the expression levels of Toll-like receptors 2 (TLR2) and 4 (TLR4) in monocytes and neutrophils from patients with KC and control subjects for demonstrating the role of innate immunity in KC. We also study the correlation between TLR2/TLR4 expression and serum levels of proinflammatory markers (IL-1β, IL-6, TNF-α, MMP-9 and NF-κB). Forty patients with bilateral KC (55% males; mean age; 33.1 ± 10.9 years) and 20 control subjects (55% males; mean age; 30.4 ± 7.6 years) were included in the study. Our results showed that mean expression of TLR2 and TLR4 in both neutrophils and monocytes was significantly higher in patients with KC compared to control subjects (all p < 0.0001). Furthermore, KC patients also showed higher serum levels of IL-1B, IL-6, TNF-α, MMP-9 (all p < 0.0001) and NF-κB (p = 0.036). In addition, we found a strong correlation between TLR2 expression in both monocytes and neutrophils (all p < 0.0001), and TLR4 in monocytes (all p < 0.05) with serum levels of IL-1B, IL-6, TNF-α and MMP-9. In conclusion, these findings suggest that TLRs may play an important role in the pathophysiology of KC.