bluuebunny/crossref_metadata_2025 huggingface dataset:
Metamaterial bricks and quantization of meta-surfaces
Authors: Memoli Gianluca , Caleap Mihai , Asakawa Michihiro , Sahoo Deepak R. , Drinkwater Bruce W. , Subramanian Sriram
Published: February 2017

Controlling acoustic fields is crucial in diverse applications such as loudspeaker design, ultrasound imaging and therapy or acoustic particle manipulation. The current approaches use fixed lenses or expensive phased arrays. Here, using a process of analogue-to-digital conversion and wavelet decomposition, we develop the notion of quantal meta-surfaces. The quanta here are small, pre-manufactured three-dimensional units—which we call metamaterial bricks—each encoding a specific phase delay. These bricks can be assembled into meta-surfaces to generate any diffraction-limited acoustic field. We apply this methodology to show experimental examples of acoustic focusing, steering and, after stacking single meta-surfaces into layers, the more complex field of an acoustic tractor beam. We demonstrate experimentally single-sided air-borne acoustic levitation using meta-layers at various bit-rates: from a 4-bit uniform to 3-bit non-uniform quantization in phase. This powerful methodology dramatically simplifies the design of acoustic devices and provides a key-step towards realizing spatial sound modulators.
Opportunities to develop the professional role of community pharmacists in the care of patients with asthma: a cross-sectional study
Authors: Watkins Kim , Bourdin Aline , Trevenen Michelle , Murray Kevin , Kendall Peter A , Schneider Carl R , Clifford Rhonda
Published: February 2017

There are many indications in Australia and globally that asthma management is suboptimal. Ideally, patients need to proactively self-manage the condition with the support of health professionals. Community pharmacists are a highly accessible resource for patients but currently provide inconsistent services. General practitioners also face many barriers to the provision of chronic disease management for asthma patients. The aim of this research was to characterise patients with asthma who present to community pharmacy. The objective was to identify opportunities to develop the role of pharmacists in the context of the primary healthcare setting and in view of the needs of the patients they routinely encounter. The results of a comprehensive survey of 248 patients recruited from community pharmacies indicated there was discordance between patient perceptions of asthma control and actual asthma control. Almost half the patients surveyed had poorly controlled asthma, whereas almost three quarters perceived their asthma to be well or completely controlled. Fewer than 20% of patients were utilising written asthma action plans, and issues around quality use of medicines were identified. The significance of the incongruent perceptions regarding asthma control is that patients are unlikely to proactively seek intervention and support from healthcare professionals. Community pharmacists provide a significant opportunity to address these issues by direct intervention. There is scope to investigate pharmacists preparing written asthma action plans for patients, using software to monitor medication adherence and prescribe on-going medication. To maximise the potential of pharmacists, barriers to practice need to be identified and addressed.
Fatty acid synthase regulates estrogen receptor-α signaling in breast cancer cells
Authors: Menendez J A , Lupu R
Published: February 2017

Fatty acid synthase (FASN), the key enzyme for endogenous synthesis of fatty acids, is overexpressed and hyperactivated in a biologically aggressive subset of sex steroid-related tumors, including breast carcinomas. Using pharmacological and genetic approaches, we assessed the molecular relationship between FASN signaling and estrogen receptor alpha (ERα) signaling in breast cancer. The small compound C75, a synthetic slow-binding inhibitor of FASN activity, induced a dramatic augmentation of estradiol (E2)-stimulated, ERα-driven transcription. FASN and ERα were both necessary for the synergistic activation of ERα transcriptional activity that occurred following co-exposure to C75 and E2: first, knockdown of FASN expression using RNAi (RNA interference) drastically lowered (>100 fold) the amount of E2required for optimal activation of ERα-mediated transcriptional activity; second, FASN blockade synergistically increased E2-stimulated ERα-mediated transcriptional activity in ERα-negative breast cancer cells stably transfected with ERα, but not in ERα-negative parental cells. Non-genomic, E2-regulated cross-talk between the ERα and MAPK pathways participated in these phenomena. Thus, treatment with the pure antiestrogen ICI 182 780 or the potent and specific inhibitor of MEK/ERK, U0126, was sufficient to abolish the synergistic nature of the interaction between FASN blockade and E2-stimulated ERα transactivation. FASN inhibition suppressed E2-stimulated breast cancer cell proliferation and anchorage-independent colony formation while promoting the reduction of ERα protein. FASN blockade resulted in the increased expression and nuclear accumulation of the cyclin-dependent kinase inhibitors p21WAF1/CIP1and p27Kip1, two critical mediators of the therapeutic effects of antiestrogen in breast cancer, while inactivating AKT, a key mediator of E2-promoted anchorage-independent growth. The ability of FASN to regulate E2/ERα signaling may represent a promising strategy for anticancer treatment involving a new generation of FASN inhibitors.
Zero-energy pinning from interactions in Majorana nanowires
Authors: Domínguez Fernando , Cayao Jorge , San-Jose Pablo , Aguado Ramón , Yeyati Alfredo Levy , Prada Elsa
Published: February 2017

Majorana zero modes at the boundaries of topological superconductors are charge-neutral, an equal superposition of electrons and holes. This ideal situation is, however, hard to achieve in physical implementations, such as proximitized semiconducting nanowires of realistic length. In such systems Majorana overlaps are unavoidable and lead to their hybridization into charged Bogoliubov quasiparticles of finite energy, which, unlike true zero modes, are affected by electronic interactions. We here demonstrate that these interactions, particularly with bound charges in the dielectric surroundings, drastically change the non-interacting paradigm. Remarkably, interactions may completely suppress Majorana hybridization around parity crossings, where the total charge in the nanowire changes. This effect, dubbed zero-energy pinning, stabilizes Majoranas back to zero energy and charge, and leads to electronically incompressible parameter regions wherein Majoranas remain insensitive to local perturbations, despite their overlap.
An individual-based model for collective cancer cell migration explains speed dynamics and phenotype variability in response to growth factors
Authors: Stichel Damian , Middleton Alistair M. , Müller Benedikt F. , Depner Sofia , Klingmüller Ursula , Breuhahn Kai , Matthäus Franziska
Published: February 2017

Collective cell migration is a common phenotype in epithelial cancers, which is associated with tumor cell metastasis and poor patient survival. However, the interplay between physiologically relevant pro-migratory stimuli and the underlying mechanical cell–cell interactions are poorly understood. We investigated the migratory behavior of different collectively migrating non-small cell lung cancer cell lines in response to motogenic growth factors (e.g. epidermal growth factor) or clinically relevant small compound inhibitors. Depending on the treatment, we observed distinct behaviors in a classical lateral migration assay involving traveling fronts, finger-shapes or the development of cellular bridges. Particle image velocimetry analysis revealed characteristic speed dynamics (evolution of the average speed of all cells in a frame) in all experiments exhibiting initial acceleration and subsequent deceleration of the cell populations. To better understand the mechanical properties of individual cells leading to the observed speed dynamics and the phenotypic differences we developed a mathematical model based on a Langevin approach. This model describes intercellular forces, random motility, and stimulation of active migration by mechanical interaction between cells. Simulations show that the model is able to reproduce the characteristic spatio-temporal speed distributions as well as most migratory phenotypes of the studied cell lines. A specific strength of the proposed model is that it identifies a small set of mechanical features necessary to explain all phenotypic and dynamical features of the migratory response of non-small cell lung cancer cells to chemical stimulation/inhibition. Furthermore, all processes included in the model can be associated with potential molecular components, and are therefore amenable to experimental validation. Thus, the presented mathematical model may help to predict which mechanical aspects involved in non-small cell lung cancer cell migration are affected by the respective therapeutic treatment.
Transcriptome analysis of bagging-treated red Chinese sand pear peels reveals light-responsive pathway functions in anthocyanin accumulation
Authors: Bai Songling , Sun Yongwang , Qian Minjie , Yang Fengxia , Ni Junbei , Tao Ruiyan , Li Lin , Shu Qun , Zhang Dong , Teng Yuanwen
Published: February 2017

Bagging is an efficient method to improve fruit colour development. This work reported a transcriptome analysis using bagging-treated red Chinese sand pear peels. In total, 8,870 differentially expressed genes were further analysed by a weighted gene co-expression network analysis and early-, middle- and late light-responsive genes were identified. An annotation analysis revealed several pathways involved in the different responsive stages. The presence of LONG HYPOCOTLY 5, CRY-DASH and a CONSTANS-like transcription factors among the early light-responsive genes indicated the pivotal role of light, especially blue light, in the biological changes that occurred after bag removal. Other light-responsive transcription factors were also identified from the three light-responsive stages. In addition, the light-responsive pattern of anthocyanin biosynthetic genes differed among the biosynthetic steps. Although yeast-one hybrid assay showed that most of the structural genes were regulated by PpMYB10, their different temporal expressive pattern suggested that besides PpMYB10, other light-responsive transcriptional factors were also involved in the regulation of anthocyanin biosynthesis. In summary, our transcriptome analysis provides knowledge of the transcriptional regulatory network operating during light responses, which results in anthocyanin accumulation and other significant physiological changes in red Chinese sand pear peels after bag removal.
Serum microRNA-1 and microRNA-133a levels reflect myocardial steatosis in uncomplicated type 2 diabetes
Authors: de Gonzalo-Calvo D. , van der Meer R. W. , Rijzewijk L. J. , Smit J. W. A. , Revuelta-Lopez E. , Nasarre L. , Escola-Gil J. C. , Lamb H. J. , Llorente-Cortes V.
Published: February 2017

Using in vitro, in vivo and patient-based approaches, we investigated the potential of circulating microRNAs (miRNAs) as surrogate biomarkers of myocardial steatosis, a hallmark of diabetic cardiomyopathy. We analysed the cardiomyocyte-enriched miRNA signature in serum from patients with well-controlled type 2 diabetes and with verified absence of structural heart disease or inducible ischemia, and control volunteers of the same age range and BMI (N = 86), in serum from a high-fat diet-fed murine model, and in exosomes from lipid-loaded HL-1 cardiomyocytes. Circulating miR-1 and miR-133a levels were robustly associated with myocardial steatosis in type 2 diabetes patients, independently of confounding factors in both linear and logistic regression analyses (P < 0.050 for all models). Similar to myocardial steatosis, miR-133a levels were increased in type 2 diabetes patients as compared with healthy subjects (P < 0.050). Circulating miR-1 and miR-133a levels were significantly elevated in high-fat diet-fed mice (P < 0.050), which showed higher myocardial steatosis, as compared with control animals. miR-1 and miR-133a levels were higher in exosomes released from lipid-loaded HL-1 cardiomyocytes (P < 0.050). Circulating miR-1 and miR-133a are independent predictors of myocardial steatosis. Our results highlight the value of circulating miRNAs as diagnostic tools for subclinical diabetic cardiomyopathy.
Quantitative Limits on Small Molecule Transport via the Electropermeome — Measuring and Modeling Single Nanosecond Perturbations
Authors: Sözer Esin B. , Levine Zachary A. , Vernier P. Thomas
Published: February 2017

The detailed molecular mechanisms underlying the permeabilization of cell membranes by pulsed electric fields (electroporation) remain obscure despite decades of investigative effort. To advance beyond descriptive schematics to the development of robust, predictive models, empirical parameters in existing models must be replaced with physics- and biology-based terms anchored in experimental observations. We report here absolute values for the uptake of YO-PRO-1, a small-molecule fluorescent indicator of membrane integrity, into cells after a single electric pulse lasting only 6 ns. We correlate these measured values, based on fluorescence microphotometry of hundreds of individual cells, with a diffusion-based geometric analysis of pore-mediated transport and with molecular simulations of transport across electropores in a phospholipid bilayer. The results challenge the “drift and diffusion through a pore” model that dominates conventional explanatory schemes for the electroporative transfer of small molecules into cells and point to the necessity for a more complex model.
RNA-Sequencing data supports the existence of novel VEGFA splicing events but not of VEGFAxxxb isoforms
Authors: Bridgett Stephen , Dellett Margaret , Simpson David A.
Published: February 2017

Vascular endothelial growth factor (VEGFA), a pivotal regulator of angiogenesis and valuable therapeutic target, is characterised by alternative splicing which generates three principal isoforms, VEGFA121, VEGFA165 and VEGFA189. A second set of anti-angiogenic isoforms termed VEGFAxxxb that utilise an alternative splice site in the final exon have been widely reported, with mRNA detection based principally upon RT-PCR assays. We sought confirmation of the existence of the VEGFAxxxb isoforms within the abundant RNA sequencing data available publicly. Whilst sequences derived specifically from each of the canonical VEGFA isoforms were present in many tissues, there were no sequences derived from VEGFAxxxb isoforms. Sequencing of approximately 50,000 RT-PCR products spanning the exon 7–8 junction in 10 tissues did not identify any VEGFAxxxb transcripts. The absence or extremely low expression of these transcripts in vivo indicates that VEGFAxxxb isoforms are unlikely to play a role in normal physiology. Our analyses also revealed multiple novel splicing events supported by more reads than previously reported for VEGFA145 and VEGFA148 isoforms, including three from novel first exons consistent with existing transcription start site data. These novel VEGFA isoforms may play significant roles in specific cell types.
Evaluation of Recombinant Human FGF-2 and PDGF-BB in Periodontal Regeneration: A Systematic Review and Meta-Analysis
Authors: Li Feifei , Yu Fanyuan , Xu Xin , Li Chunjie , Huang Dingming , Zhou Xuedong , Ye Ling , Zheng Liwei
Published: February 2017

The prognosis for successful treatment of periodontal diseases is generally poor. Current therapeutic strategies often fail to regenerate infected periodontium. Recently an alternative strategy has been developed that combines conventional treatment with the application of recombinant human growth factors (rhGFs). But ambiguities in existed studies on the clinical efficacy of rhGFs do not permit either the identification of the specific growth factors effective for therapeutic interventions or the optimal concentration of them. Neither is it known whether the same rhGF can stimulate regeneration of both soft tissue and bone, or whether different patient populations call for differential use of the growth factors. In order to explore these issues, a meta-analysis was carried out. Particular attention was given to the therapeutic impact of fibroblast growth factor 2(FGF-2) and platelet derived growth factor BB (PDGF-BB). Our findings indicate that 0.3% rhFGF-2 and 0.3 mg/ml rhPDGF-BB show a greater capacity for periodontal regeneration than other concentrations and superiority to control groups with statistical significance. In the case of patients suffering only from gingival recession, however, the application of rhPDGF-BB produces no significant regenerative advantage. The findings of this study can potentially endow clinicians with guidelines for the appropriate application of these two rhGFs.
Mycoplasma bovis NADH oxidase functions as both a NADH oxidizing and O2 reducing enzyme and an adhesin
Authors: Zhao Gang , Zhang Hui , Chen Xi , Zhu Xifang , Guo Yusi , He Chenfei , Anwar Khan Farhan , Chen Yingyu , Hu Changmin , Chen Huanchun , Guo Aizhen
Published: February 2017

Mycoplasma bovis causes considerable economic losses in the cattle industry worldwide. In mycoplasmal infections, adhesion to the host cell is of the utmost importance. In this study, the amino acid sequence of NOX was predicted to have enzymatic domains. The nox gene was then cloned and expressed in Escherichia coli. The enzymatic activity of recombinant NOX (rNOX) was confirmed based on its capacity to oxidize NADH to NAD+ and reduce O2 to H2O2. The adherence of rNOX to embryonic bovine lung (EBL) cells was confirmed with confocal laser scanning microscopy, enzyme-linked immunosorbent assay, and flow cytometry. Both preblocking EBL cells with purified rNOX and preneutralizing M. bovis with polyclonal antiserum to rNOX significantly reduced the adherence of M. bovis to EBL cells. Mycoplasma bovisNOX–expressed a truncated NOX protein at a level 10-fold less than that of the wild type. The capacities of M. bovisNOX– for cell adhesion and H2O2 production were also significantly reduced. The rNOX was further used to pan phage displaying lung cDNA library and fibronectin was determined to be potential ligand. In conclusion, M. bovis NOX functions as both an active NADH oxidase and adhesin, and is therefore a potential virulence factor.
A c-Myc-regulated stem cell-like signature in high-risk neuroblastoma: A systematic discovery (Target neuroblastoma ESC-like signature)
Authors: Yang Xinan , Tang Fangming , Shin Jisu , Cunningham John M.
Published: February 2017

c-Myc dysregulation is hypothesized to account for the ‘stemness’ – self-renewal and pluripotency – shared between embryonic stem cells (ESCs) and adult aggressive tumours. High-risk neuroblastoma (HR-NB) is the most frequent, aggressive, extracranial solid tumour in childhood. Using HR-NB as a platform, we performed a network analysis of transcriptome data and presented a c-Myc subnetwork enriched for genes previously reported as ESC-like cancer signatures. A subsequent drug-gene interaction analysis identified a pharmacogenomic agent that preferentially interacted with this HR-NB-specific, ESC-like signature. This agent, Roniciclib (BAY 1000394), inhibited neuroblastoma cell growth and induced apoptosis in vitro. It also repressed the expression of the oncogene c-Myc and the neural ESC marker CDK2 in vitro, which was accompanied by altered expression of the c-Myc-targeted cell cycle regulators CCND1, CDKN1A and CDKN2D in a time-dependent manner. Further investigation into this HR-NB-specific ESC-like signature in 295 and 243 independent patients revealed and validated the general prognostic index of CDK2 and CDKN3 compared with CDKN2D and CDKN1B. These findings highlight the very potent therapeutic benefits of Roniciclib in HR-NB through the targeting of c-Myc-regulated, ESC-like tumorigenesis. This work provides a hypothesis-driven systems computational model that facilitates the translation of genomic and transcriptomic signatures to molecular mechanisms underlying high-risk tumours.
Differential expression of the protein kinase A subunits in normal adrenal glands and adrenocortical adenomas
Authors: Weigand Isabel , Ronchi Cristina L. , Rizk-Rabin Marthe , Dalmazi Guido Di , Wild Vanessa , Bathon Kerstin , Rubin Beatrice , Calebiro Davide , Beuschlein Felix , Bertherat Jérôme , Fassnacht Martin , Sbiera Silviu
Published: February 2017

Somatic mutations in protein kinase A catalytic α subunit (PRKACA) were found to be causative for 30–40% of cortisol-producing adenomas (CPA) of the adrenal gland, rendering PKA signalling constitutively active. In its resting state, PKA is a stable and inactive heterotetramer, consisting of two catalytic and two regulatory subunits with the latter inhibiting PKA activity. The human genome encodes three different PKA catalytic subunits and four different regulatory subunits that are preferentially expressed in different organs. In normal adrenal glands all regulatory subunits are expressed, while CPA exhibit reduced protein levels of the regulatory subunit IIβ. In this study, we linked for the first time the loss of RIIβ protein levels to the PRKACA mutation status and found the down-regulation of RIIβ to arise post-transcriptionally. We further found the PKA subunit expression pattern of different tumours is also present in the zones of the normal adrenal cortex and demonstrate that the different PKA subunits have a differential expression pattern in each zone of the normal adrenal gland, indicating potential specific roles of these subunits in the regulation of different hormones secretion.
Usp7-dependent histone H3 deubiquitylation regulates maintenance of DNA methylation
Authors: Yamaguchi Luna , Nishiyama Atsuya , Misaki Toshinori , Johmura Yoshikazu , Ueda Jun , Arita Kyohei , Nagao Koji , Obuse Chikashi , Nakanishi Makoto
Published: February 2017

Uhrf1-dependent histone H3 ubiquitylation plays a crucial role in the maintenance of DNA methylation via the recruitment of the DNA methyltransferase Dnmt1 to DNA methylation sites. However, the involvement of deubiquitylating enzymes (DUBs) targeting ubiquitylated histone H3 in the maintenance of DNA methylation is largely unknown. With the use of Xenopus egg extracts, we demonstrate here that Usp7, a ubiquitin carboxyl-terminal hydrolase, forms a stable complex with Dnmt1 and is recruited to DNA methylation sites during DNA replication. Usp7 deubiquitylates ubiquitylated histone H3 in vitro. Inhibition of Usp7 activity or its depletion in egg extracts results in enhanced and extended binding of Dnmt1 to chromatin, suppressing DNA methylation. Depletion of Usp7 in HeLa cells causes enhanced histone H3 ubiquitylation and enlargement of Dnmt1 nuclear foci during DNA replication. Our results thus suggest that Usp7 is a key factor that regulates maintenance of DNA methylation.
Serum resistin is causally related to mortality risk in patients with type 2 diabetes: preliminary evidences from genetic data
Authors: Fontana Andrea , Ortega Moreno Lorena , Lamacchia Olga , De Bonis Concetta , Salvemini Lucia , De Cosmo Salvatore , Cignarelli Mauro , Copetti Massimiliano , Trischitta Vincenzo , Menzaghi Claudia
Published: February 2017

Resistin has been firmly associated with all-cause mortality. We investigated, whether, in patients with type 2 diabetes (T2D), this association is sustained by a cause-effect relationship. A genotype risk score (GRS), created by summing the number of resistin increasing alleles of two genome-wide association studies (GWAS)-derived single nucleotide polymorphisms (SNPs), serum resistin measurements and all-cause death records were obtained in 1,479 (403 events/12,454 person-years), patients with T2D from three cohorts, Gargano Heart Study-prospective design (n = 350), Gargano Mortality Study (n = 698) and Foggia Mortality Study (n = 431), from Italy. GRS was strongly associated with serum resistin in a non-linear fashion (overall p = 3.5 * 10−7) with effect size modest for GRS = 1 and 2 and much higher for GRS >3, with respect to GRS = 0. A significant non-linear association was observed also between GRS and all-cause mortality (overall p = 3.3 * 10−2), with a low effect size for GRS = 1 and 2, and nearly doubled for GRS ≥ 3, with respect to GRS = 0. Based on the above-reported associations, each genetic equivalent SD increase in log-resistin levels showed a causal hazard ratio of all-cause mortality equal to 2.17 (95%CI: 1.22–3.87), thus providing evidence for a causal role of resistin in shaping the risk of mortality in diabetic patients.
Rescue of the MERTK phagocytic defect in a human iPSC disease model using translational read-through inducing drugs
Authors: Ramsden Conor M. , Nommiste Britta , R. Lane Amelia , Carr Amanda-Jayne F. , Powner Michael B. , J. K. Smart Matthew , Chen Li Li , Muthiah Manickam N. , Webster Andrew R. , Moore Anthony T. , Cheetham Michael E. , da Cruz Lyndon , Coffey Peter J.
Published: February 2017

Inherited retinal dystrophies are an important cause of blindness, for which currently there are no effective treatments. In order to study this heterogeneous group of diseases, adequate disease models are required in order to better understand pathology and to test potential therapies. Induced pluripotent stem cells offer a new way to recapitulate patient specific diseases in vitro, providing an almost limitless amount of material to study. We used fibroblast-derived induced pluripotent stem cells to generate retinal pigment epithelium (RPE) from an individual suffering from retinitis pigmentosa associated with biallelic variants in MERTK. MERTK has an essential role in phagocytosis, one of the major functions of the RPE. The MERTK deficiency in this individual results from a nonsense variant and so the MERTK-RPE cells were subsequently treated with two translational readthrough inducing drugs (G418 & PTC124) to investigate potential restoration of expression of the affected gene and production of a full-length protein. The data show that PTC124 was able to reinstate phagocytosis of labeled photoreceptor outer segments at a reduced, but significant level. These findings represent a confirmation of the usefulness of iPSC derived disease specific models in investigating the pathogenesis and screening potential treatments for these rare blinding disorders.
Three-dimensional mouse brain cytoarchitecture revealed by laboratory-based x-ray phase-contrast tomography
Authors: Töpperwien Mareike , Krenkel Martin , Vincenz Daniel , Stöber Franziska , Oelschlegel Anja M. , Goldschmidt Jürgen , Salditt Tim
Published: February 2017

Studies of brain cytoarchitecture in mammals are routinely performed by serial sectioning of the specimen and staining of the sections. The procedure is labor-intensive and the 3D architecture can only be determined after aligning individual 2D sections, leading to a reconstructed volume with non-isotropic resolution. Propagation-based x-ray phase-contrast tomography offers a unique potential for high-resolution 3D imaging of intact biological specimen due to the high penetration depth and potential resolution. We here show that even compact laboratory CT at an optimized liquid-metal jet microfocus source combined with suitable phase-retrieval algorithms and a novel tissue preparation can provide cellular and subcellular resolution in millimeter sized samples of mouse brain. We removed water and lipids from entire mouse brains and measured the remaining dry tissue matrix in air, lowering absorption but increasing phase contrast. We present single-cell resolution images of mouse brain cytoarchitecture and show that axons can be revealed in myelinated fiber bundles. In contrast to optical 3D techniques our approach does neither require staining of cells nor tissue clearing, procedures that are increasingly difficult to apply with increasing sample and brain sizes. The approach thus opens a novel route for high-resolution high-throughput studies of brain architecture in mammals.
A comparison of host gene expression signatures associated with infection in vitro by the Makona and Ecran (Mayinga) variants of Ebola virus
Authors: Bosworth Andrew , Dowall Stuart D. , Garcia-Dorival Isabel , Rickett Natasha Y. , Bruce Christine B. , Matthews David A. , Fang Yongxiang , Aljabr Waleed , Kenny John , Nelson Charlotte , Laws Thomas R. , Williamson E. Diane , Stewart James P. , Carroll Miles W. , Hewson Roger , Hiscox Julian A.
Published: February 2017

The Ebola virus (EBOV) variant Makona (which emerged in 2013) was the causative agent of the largest outbreak of Ebola Virus Disease recorded. Differences in virus-host interactions between viral variants have potential consequences for transmission, disease severity and mortality. A detailed profile of the cellular changes induced by the Makona variant compared with other Ebola virus variants was lacking. In this study, A549 cells, a human cell line with a robust innate response, were infected with the Makona variant or with the Ecran variant originating from the 1976 outbreak in Central Africa. The abundance of viral and cellular mRNA transcripts was profiled using RNASeq and differential gene expression analysis performed. Differences in effects of each virus on the expression of interferon-stimulated genes were also investigated in A549 NPro cells where the type 1 interferon response had been attenuated. Cellular transcriptomic changes were compared with those induced by human respiratory syncytial virus (HRSV), a virus with a similar genome organisation and replication strategy to EBOV. Pathway and gene ontology analysis revealed differential expression of functionally important genes; including genes involved in the inflammatory response, cell proliferation, leukocyte extravasation and cholesterol biosynthesis. Whilst there was overlap with HRSV, there was unique commonality to the EBOV variants.
Calcium pre-conditioning substitution enhances viability and glucose sensitivity of pancreatic beta-cells encapsulated using polyelectrolyte multilayer coating method
Authors: Nikravesh Niusha , Cox Sophie C. , Birdi Gurpreet , Williams Richard L. , Grover Liam M.
Published: February 2017

Type I diabetics are dependent on daily insulin injections. A therapy capable of immunoisolating pancreatic beta-cells and providing normoglycaemia is an alternative since it would avoid the late complications associated with insulin use. Here, 3D-concave agarose micro-wells were used to culture robust pancreatic MIN-6 cell spheroids within 24 hours that were shown to exhibit cell-cell contact and uniform size (201 ± 2 μm). A polyelectrolyte multilayer (PEM) approach using alginate and poly-l-lysine was employed to coat cell spheroids. In comparison to conventional PEM, use of a novel Ca2+ pre-coating step enhanced beta-cells viability (89 ± 6%) and metabolic activity since it reduced the toxic effect of the cationic polymer. Pre-coating was achieved by treating MIN-6 spheroids with calcium chloride, which enabled the adhesion of anionic polymer to the cells surface. Pre-coated cells coated with four bilayers of polymers were successfully immunoisolated from FITC-mouse antibody and pro-inflammatory cytokines. Novel PEM coated cells were shown to secret significantly (P < 0.05) different amounts of insulin in response to changes in glucose concentration (2 vs. 20 mM). This work presents a 3D culture model and novel PEM coating procedure that enhances viability, maintains functionality and immunoisolates beta-cells, which is a promising step towards an alternative therapy to insulin.
Venomics of Tropidolaemus wagleri, the sexually dimorphic temple pit viper: Unveiling a deeply conserved atypical toxin arsenal
Authors: Tan Choo Hock , Tan Kae Yi , Yap Michelle Khai Khun , Tan Nget Hong
Published: February 2017

Tropidolaemus wagleri (temple pit viper) is a medically important snake in Southeast Asia. It displays distinct sexual dimorphism and prey specificity, however its venomics and inter-sex venom variation have not been thoroughly investigated. Applying reverse-phase HPLC, we demonstrated that the venom profiles were not significantly affected by sex and geographical locality (Peninsular Malaya, insular Penang, insular Sumatra) of the snakes. Essentially, venoms of both sexes share comparable intravenous median lethal dose (LD50) (0.56–0.63 μg/g) and cause neurotoxic envenomation in mice. LCMS/MS identified six waglerin forms as the predominant lethal principles, comprising 38.2% of total venom proteins. Fourteen other toxin-protein families identified include phospholipase A2, serine proteinase, snaclec and metalloproteinase. In mice, HPLC fractions containing these proteins showed insignificant contribution to the overall venom lethality. Besides, the unique elution pattern of approximately 34.5% of non-lethal, low molecular mass proteins (3–5 kDa) on HPLC could be potential biomarker for this primitive crotalid species. Together, the study unveiled the venom proteome of T. wagleri that is atypical among many pit vipers as it comprises abundant neurotoxic peptides (waglerins) but little hemotoxic proteinases. The findings also revealed that the venom is relatively well conserved intraspecifically despite the drastic morphological differences between sexes.
A longitudinal study of cerebral blood flow under hypoxia at high altitude using 3D pseudo-continuous arterial spin labeling
Authors: Liu Wenjia , Liu Jie , Lou Xin , Zheng Dandan , Wu Bing , Wang Danny J. J. , Ma Lin
Published: February 2017

Changes in cerebral blood flow (CBF) may occur with acute exposure to high altitude; however, the CBF of the brain parenchyma has not been studied to date. In this study, identical magnetic resonance scans using arterial spin labeling (ASL) were performed to study the haemodynamic changes at both sea level and high altitude. We found that with acute exposure to high altitude, the CBF in acute mountain sickness (AMS) subjects was higher (P < 0.05), while the CBF of non-AMS subjects was lower (P > 0.05) compared with those at sea level. Moreover, magnetic resonance angiography in both AMS and non-AMS subjects showed a significant increase in the cross-sectional areas of the internal carotid, basilar, and middle cerebral arteries on the first day at high altitude. These findings support that AMS may be related to increased CBF rather than vasodilation; these results contradict most previous studies that reported no relationship between CBF changes and the occurrence of AMS. This discrepancy may be attributed to the use of ASL for CBF measurement at both sea level and high altitude in this study, which has substantial advantages over transcranial Doppler for the assessment of CBF.
Thermoelectric SnS and SnS-SnSe solid solutions prepared by mechanical alloying and spark plasma sintering: Anisotropic thermoelectric properties
Authors: Asfandiyar  , Wei Tian-Ran , Li Zhiliang , Sun Fu-Hua , Pan Yu , Wu Chao-Feng , Farooq Muhammad Umer , Tang Huaichao , Li Fu , Li Bo , Li Jing-Feng
Published: February 2017

P–type SnS compound and SnS1−xSex solid solutions were prepared by mechanical alloying followed by spark plasma sintering (SPS) and their thermoelectric properties were then studied in different compositions (x = 0.0, 0.2, 0.5, 0.8) along the directions parallel (//) and perpendicular (⊥) to the SPS–pressurizing direction in the temperature range 323–823 Κ. SnS compound and SnS1−xSex solid solutions exhibited anisotropic thermoelectric performance and showed higher power factor and thermal conductivity along the direction ⊥ than the // one. The thermal conductivity decreased with increasing contents of Se and fell to 0.36 W m−1 K−1 at 823 K for the composition SnS0.5Se0.5. With increasing selenium content (x) the formation of solid solutions substantially improved the electrical conductivity due to the increased carrier concentration. Hence, the optimized power factor and reduced thermal conductivity resulted in a maximum ZT value of 0.64 at 823 K for SnS0.2Se0.8 along the parallel direction.
Exogenous melatonin reduces somatic cell count of milk in Holstein cows
Authors: Yang Minghui , Shi Jianmin , Tian Jianhua , Tao Jingli , Chai Menglong , Wang Jing , Xu Zhiyuan , Song Yukun , Zhu Kuanfeng , Ji Pengyun , Liu Guoshi
Published: February 2017

High somatic cell counts in milk caused by mastitis significantly influence the quality of milk and result in substantial annual economic loss. This study evaluated the beneficial effects of melatonin (MT) on milk somatic cell count (SCC) in cows. To examine the effects of melatonin on SCC, one hundred twenty cows were divided into four groups based on milk SCC. In each group, half of the cows were treated with melatonin (S.C.). Melatonin treatment significantly reduced milk SCC. To explore the potential mechanism, 20 cows with relatively high SCC were selected to evaluate the biochemical and immunological profiles of their blood after melatonin treatment. Treatment with MT significantly reduced SCC in milk, lowered serum cortisol concentrations and increased the levels of albumin, alanine transaminase and lactate dehydrogenase. Following treatment with MT, the concentration of IgG and IgM rose transiently then decreased significantly, similar to changes observed for white blood cells and lymphocytes. In conclusion, MT treatment improved the quality of milk by reducing SCC. This may be due to melatonin improving immune activity in cows.
The influences of four types of soil on the growth, physiological and biochemical characteristics of Lycoris aurea (L’ Her.) Herb
Authors: Quan Miaohua , Liang Juan
Published: February 2017

Based on the characteristics of Lycoris aurea (L. aurea) natural distribution and local soil types, we selected four representative types of soil, including humus soil, sandy soil, garden soil and yellow-brown soil, for conducting the cultivation experiments to investigate key soil factors influencing its growth and development and to select the soil types suitable for cultivating it. We found that there existed significant differences in the contents of mineral elements and the activities of soil enzymes (urease, phosphatase, sucrase and catalase) etc. Among which, the contents of organic matters, alkali-hydrolysable nitrogen, Ca and Mg as well as the activities of soil enzymes in humus soil were the highest ones. In yellow-brown soil, except for Fe, the values of all the other items were the lowest ones. Net photosynthetic rate (Pn), biomass and lycorine content in humus soil were all the highest ones, which were increased by 31.02, 69.39 and 55.79%, respectively, as compared to those of yellow-brown soil. Stepwise multiple regression analysis and path analysis indicated that alkali-hydrolysable nitrogen, and Ca etc. were key soil factors influencing Pn, biomass and lycorine content of L. aurea. Thus, humus soil can be used as medium suitable for artificial cultivation of L. aurea.
Association of Renal Elasticity and Renal Function Progression in Patients with Chronic Kidney Disease Evaluated by Real-Time Ultrasound Elastography
Authors: Lin Hugo You-Hsien , Lee Yu-Li , Lin Kun-Der , Chiu Yi-Wen , Shin Shyi-Jang , Hwang Shang-Jyh , Chen Hung-Chun , Hung Chi-Chih
Published: February 2017

Glomerulosclerosis and tubulointerstitial fibrosis are associated with lower renal parenchymal elasticity. This study was designed to evaluate the predictive ability of renal elasticity in patients with chronic kidney disease (CKD). 148 non-CKD patients and 227 patients with CKD were recruited. 145 (38.7%) were female, 166 (73.1%) had diabetes, the mean estimated glomerular filtration rate (eGFR) was 33.9 ± 15.8 ml/min/1.73 m2 and the median urinary protein-to-creatinine ratio (UPCR) 502 (122–1491) mg/g. Patients with later stages of CKD had lower renal elasticity values, indicating stiffer kidneys (p < 0.001), and smaller kidney (p < 0.001). Renal elasticity correlated with log-transformed UPCR (β = −7.544, P < 0.001). Renal length correlated with age (β = −0.231, P < 0.001), sex (β = −3.730, P < 0.001), serum albumin level (β = −3.024, P = 0.001), body mass index (β = 0.390, P = 0.009) and eGFR (β = 0.146, P < 0.001). In fully-adjusted logistic regression model, the odds ratio (OR) per 10 unit change in renal elasticity for rapid renal deterioration was 0.928 (95% CI, 0.864–0.997; P = 0.042). The OR per 1 mm change in renal length for rapid renal deterioration was 1.022 (95% CI, 0.994–1.050; P = 0.125). Renal elasticity is associated with proteinuria and rapid renal deterioration in patients with CKD.
Characterization of the CD177 interaction with the ANCA antigen proteinase 3
Authors: Jerke Uwe , Marino Stephen F. , Daumke Oliver , Kettritz Ralph
Published: February 2017

Proteinase 3 is a serine protease found in neutrophil granules and on the extracellular neutrophil membrane (mPR3). mPR3 is a major antigen for anti-neutrophil cytoplasmic antibodies (PR3-ANCAs), autoantibodies causing fatal autoimmune diseases. In most individuals, a subpopulation of neutrophils also produce CD177, proposed to present additional PR3 on the surface, resulting in CD177neg/mPR3low and CD177pos/mPR3high neutrophil subsets. A positive correlation has been shown between mPR3 abundance, disease incidence, and clinical outcome. We present here a detailed investigation of the PR3:CD177 complex, verifying the interaction, demonstrating the effect of binding on PR3 proteolytic activity and explaining the accessibility of major PR3-ANCA epitopes. We observed high affinity PR3:CD177 complex formation by surface plasmon resonance. Using flow cytometry and a PR3-specific FRET assay, we found that CD177 binding reduced the proteolytic activity of PR3 in vitro using purified proteins, in neutrophil degranulation supernatants containing wtPR3 and directly on mPR3high neutrophils and PR3-loaded HEK cells. Finally, CD177pos/mPR3high neutrophils showed no migration advantage in vitro or in vivo when migrating from the blood into the oral cavity. We illuminate details of the PR3:CD177 interaction explaining mPR3 membrane orientation and proteolytic activity with relevance to ANCA activation of the distinct mPR3 neutrophil populations.
Impact of radiation attenuation by a carbon fiber couch on patient dose verification
Authors: Yu Chun-Yen , Chou Wen-Tsae , Liao Yi-Jen , Lee Jeng-Hung , Liang Ji-An , Hsu Shih-Ming
Published: February 2017

The aim of this study was to understand the difference between the measured and calculated irradiation attenuations obtained using two algorithms and to identify the influence of couch attenuation on patient dose verification. We performed eight tests of couch attenuation with two photon energies, two longitudinal couch positions, and two rail positions. The couch attenuation was determined using a radiation treatment planning system. The measured and calculated attenuations were compared. We also performed 12 verifications of head-and-neck and rectum cases by using a Delta phantom. The dose deviation (DD), distance to agreement (DTA), and gamma index of pencil-beam convolution (PBC) verifications were nearly the same. The agreement was least consistent for the anisotropic analytical algorithm (AAA) without the couch for the head-and-neck case, in which the DD, DTA, and gamma index were 74.4%, 99.3%, and 89%, respectively; for the rectum case, the corresponding values were 56.2%, 95.1%, and 92.4%. We suggest that dose verification should be performed using the following three metrics simultaneously: DD, DTA, and the gamma index.
Enhanced cytomegalovirus infection in human trabecular meshwork cells and its implication in glaucoma pathogenesis
Authors: Choi Jin A , Kim Ju-Eun , Noh Seung-Jun , Kyoung Kim Eun , Park Chan Kee , Paik Soon-Young
Published: February 2017

Cytomegalovirus (CMV) is one of the infectious causes of hypertensive anterior uveitis, which is characterized by recurrent episodes of elevated intraocular pressure (IOP) and mild anterior uveitis. Despite the potentially vision-threatening complications of this disease, the underlying mechanisms remain largely undefined. We aimed to investigate whether human trabecular meshwork (TM) cells, the key cell type that regulates IOP, could support CMV replication, as well as demonstrate the relevant pathological changes in TM. When human TM cells were infected with CMV AD169, immediate early antigens were detected 1 day post-infection (dpi); cytopathic changes including rounding, a ballooned appearance with disorganization, and a decreased number of stress fibers were noted in TM cells. The marked increase in viral DNA accumulation was observed most notably at 5 and 7 dpi, suggesting that the active viral infection in human TM cells could be the key mechanism underlying the elevation of IOP in anterior viral uveitis. Notably, CMV infection enhanced the production of transforming growth factor (TGF)-β1, an upstream molecule that increases the resistance of the outflow pathway in human TM cells. The increase of TGF-β1 was countervailed by additional treatment with corticosteroids. Our results provide a pathogenic mechanism for IOP elevation in viral anterior uveitis.
Novel quantitative pigmentation phenotyping enhances genetic association, epistasis, and prediction of human eye colour
Authors: Wollstein Andreas , Walsh Susan , Liu Fan , Chakravarthy Usha , Rahu Mati , Seland Johan H. , Soubrane Gisèle , Tomazzoli Laura , Topouzis Fotis , Vingerling Johannes R. , Vioque Jesus , Böhringer Stefan , Fletcher Astrid E. , Kayser Manfred
Published: February 2017

Success of genetic association and the prediction of phenotypic traits from DNA are known to depend on the accuracy of phenotype characterization, amongst other parameters. To overcome limitations in the characterization of human iris pigmentation, we introduce a fully automated approach that specifies the areal proportions proposed to represent differing pigmentation types, such as pheomelanin, eumelanin, and non-pigmented areas within the iris. We demonstrate the utility of this approach using high-resolution digital eye imagery and genotype data from 12 selected SNPs from over 3000 European samples of seven populations that are part of the EUREYE study. In comparison to previous quantification approaches, (1) we achieved an overall improvement in eye colour phenotyping, which provides a better separation of manually defined eye colour categories. (2) Single nucleotide polymorphisms (SNPs) known to be involved in human eye colour variation showed stronger associations with our approach. (3) We found new and confirmed previously noted SNP-SNP interactions. (4) We increased SNP-based prediction accuracy of quantitative eye colour. Our findings exemplify that precise quantification using the perceived biological basis of pigmentation leads to enhanced genetic association and prediction of eye colour. We expect our approach to deliver new pigmentation genes when applied to genome-wide association testing.