bluuebunny/crossref_metadata_2025 huggingface dataset:
Extended Distributed State Estimation: A Detection Method against Tolerable False Data Injection Attacks in Smart Grids
Authors: Wang Dai , Guan Xiaohong , Liu Ting , Gu Yun , Shen Chao , Xu Zhanbo
Published: March 2014

False data injection (FDI) is considered to be one of the most dangerous cyber-attacks in smart grids, as it may lead to energy theft from end users, false dispatch in the distribution process, and device breakdown during power generation. In this paper, a novel kind of FDI attack, named tolerable false data injection (TFDI), is constructed. Such attacks exploit the traditional detector’s tolerance of observation errors to bypass the traditional bad data detection. Then, a method based on extended distributed state estimation (EDSE) is proposed to detect TFDI in smart grids. The smart grid is decomposed into several subsystems, exploiting graph partition algorithms. Each subsystem is extended outward to include the adjacent buses and tie lines, and generate the extended subsystem. The Chi-squares test is applied to detect the false data in each extended subsystem. Through decomposition, the false data stands out distinctively from normal observation errors and the detection sensitivity is increased. Extensive TFDI attack cases are simulated in the Institute of Electrical and Electronics Engineers (IEEE) 14-, 39-, 118- and 300-bus systems. Simulation results show that the detection precision of the EDSE-based method is much higher than that of the traditional method, while the proposed method significantly reduces the associated computational costs.
Intratumoral Decorin Gene Delivery by AAV Vector Inhibits Brain Glioblastomas and Prolongs Survival of Animals by Inducing Cell Differentiation
Authors: Ma Hsin-I , Hueng Dueng-Yuan , Shui Hao-Ai , Han Jun-Ming , Wang Chi-Hsien , Lai Ying-Hsiu , Cheng Shi-Yuan , Xiao Xiao , Chen Ming-Teh , Yang Yi-Ping
Published: March 2014

Glioblastoma multiforme (GBM) is the most malignant cancer in the central nervous system with poor clinical prognosis. In this study, we investigated the therapeutic effect of an anti-cancer protein, decorin, by delivering it into a xenograft U87MG glioma tumor in the brain of nude mice through an adeno-associated viral (AAV2) gene delivery system. Decorin expression from the AAV vector in vitro inhibited cultured U87MG cell growth by induction of cell differentiation. Intracranial injection of AAV-decorin vector to the glioma-bearing nude mice in vivo significantly suppressed brain tumor growth and prolonged survival when compared to control non-treated mice bearing the same U87MG tumors. Proteomics analysis on protein expression profiles in the U87MG glioma cells after AAV-mediated decorin gene transfer revealed up- and down-regulation of important proteins. Differentially expressed proteins between control and AAV-decorin-transduced cells were identified through MALDI-TOF MS and database mining. We found that a number of important proteins that are involved in apoptosis, transcription, chemotherapy resistance, mitosis, and fatty acid metabolism have been altered as a result of decorin overexpression. These findings offer valuable insight into the mechanisms of the anti-glioblastoma effects of decorin. In addition, AAV-mediated decorin gene delivery warrants further investigation as a potential therapeutic approach for brain tumors.
Carvacrol and trans-Cinnamaldehyde Reduce Clostridium difficile Toxin Production and Cytotoxicity in Vitro
Authors: Mooyottu Shankumar , Kollanoor-Johny Anup , Flock Genevieve , Bouillaut Laurent , Upadhyay Abhinav , Sonenshein Abraham , Venkitanarayanan Kumar
Published: March 2014

Clostridium difficile is a nosocomial pathogen that causes a serious toxin-mediated enteric disease in humans. Reducing C. difficile toxin production could significantly minimize its pathogenicity and improve disease outcomes in humans. This study investigated the efficacy of two, food-grade, plant-derived compounds, namely trans-cinnamaldehyde (TC) and carvacrol (CR) in reducing C. difficile toxin production and cytotoxicity in vitro. Three hypervirulent C. difficile isolates were grown with or without the sub-inhibitory concentrations of TC or CR, and the culture supernatant and the bacterial pellet were collected for total toxin quantitation, Vero cell cytotoxicity assay and RT-qPCR analysis of toxin-encoding genes. The effect of CR and TC on a codY mutant and wild type C. difficile was also investigated. Carvacrol and TC substantially reduced C. difficile toxin production and cytotoxicity on Vero cells. The plant compounds also significantly down-regulated toxin production genes. Carvacrol and TC did not inhibit toxin production in the codY mutant of C. difficile, suggesting a potential codY-mediated anti-toxigenic mechanism of the plant compounds. The antitoxigenic concentrations of CR and TC did not inhibit the growth of beneficial gut bacteria. Our results suggest that CR and TC could potentially be used to control C. difficile, and warrant future studies in vivo.
Low Concentration Fe-Doped Alumina Catalysts Using Sol-Gel and Impregnation Methods: The Synthesis, Characterization and Catalytic Performance during the Combustion of Trichloroethylene
Authors: Maldonado Carolina , De la Rosa Javier , Lucio-Ortiz Carlos , Hernández-Ramírez Aracely , Barraza Felipe , Valente Jaime
Published: March 2014

The role of iron in two modes of integration into alumina catalysts was studied at 0.39 wt% Fe and tested in trichloroethylene combustion. One modified alumina was synthesized using the sol-gel method with Fe added in situ during hydrolysis; another modification was performed using calcined alumina, prepared using the sol-gel method and impregnated with Fe. Several characterization techniques were used to study the level of Fe modification in the γ-Al2O3 phase formed and to correlate the catalytic properties during trichloroethylene (TCE) combustion. The introduction of Fe in situ during the sol-gel process influenced the crystallite size, and three iron species were generated, namely, magnetite, maghemite and hematite. The impregnated Fe-alumina formed hematite and maghemite, which were highly dispersed on the γ-Al2O3 surface. The X-ray photoelectron spectra (XPS), FT-IR and Mössbauer spectroscopy analyses revealed how Fe interacted with the γ-Al2O3 lattice in both catalysts. The impregnated Fe-catalyst showed the best catalytic performance compared to the catalyst that was Fe-doped in situ by the sol-gel method; both had better catalytic activity than pure alumina. This difference in activity was correlated with the accessibility of the reactants to the hematite iron species on the surface. The chlorine poisoning for all three catalysts was less than 1.8%.
A Technology Platform to Test the Efficacy of Purification of Alginate
Authors: Paredes-Juarez Genaro , De Haan Bart , Faas Marijke , De Vos Paul
Published: March 2014

Alginates are widely used in tissue engineering technologies, e.g., in cell encapsulation, in drug delivery and various immobilization procedures. The success rates of these studies are highly variable due to different degrees of tissue response. A cause for this variation in success is, among other factors, its content of inflammatory components. There is an urgent need for a technology to test the inflammatory capacity of alginates. Recently, it has been shown that pathogen-associated molecular patterns (PAMPs) in alginate are potent immunostimulatories. In this article, we present the design and evaluation of a technology platform to assess (i) the immunostimulatory capacity of alginate or its contaminants, (ii) where in the purification process PAMPs are removed, and (iii) which Toll-like receptors (TLRs) and ligands are involved. A THP1 cell-line expressing pattern recognition receptors (PRRs) and the co-signaling molecules CD14 and MD2 was used to assess immune activation of alginates during the different steps of purification of alginate. To determine if this activation was mediated by TLRs, a THP1-defMyD88 cell-line was applied. This cell-line possesses a non-functional MyD88 coupling protein, necessary for activating NF-κB via TLRs. To identify the specific TLRs being activated by the PAMPs, we use different human embryonic kidney (HEK) cell-line that expresses only one specific TLR. Finally, specific enzyme-linked immunosorbent assays (ELISAs) were applied to identify the specific PAMP. By applying this three-step procedure, we can screen alginate in a manner, which is both labor and cost efficient. The efficacy of the platform was evaluated with an alginate that did not pass our quality control. We demonstrate that this alginate was immunostimulatory, even after purification due to reintroduction of the TLR5 activating flagellin. In addition, we tested two commercially available purified alginates. Our experiments show that these commercial alginates contained peptidoglycan, lipoteichoic acid, flagellin, and even lipopolysaccharides (LPS). The platform presented here can be used to evaluate the efficacy of purification procedures in removing PAMPs from alginates in a cost-efficient manner.
Pharmacokinetic Study in Mice of Galphimine-A, an Anxiolytic Compound from Galphimia glauca
Authors: Vargas Rodolfo , Zamilpa Alejandro , Aguilar Francisco , Herrera-Ruiz Maribel , Tortoriello Jaime , Jiménez-Ferrer Enrique
Published: March 2014

The aim of this study was to obtain pharmacokinetic data for the anxiolytic compound galphimine-A (G–A) from Galphimia glauca. G–A is the most abundant anxiolytic compound in this plant, while Galphimine-E (G–E) is the most abundant galphimine, but inactive. G–E was transformed chemically into G–A. The pharmacokinetic study was carried out in ICR mice, which were orally administered a single 200 mg/kg dose of G–A. Samples of blood and brain were taken at different times after administration of G–A. Previously, we established the validation of methods for determining the concentration of G–A. The G–A was detected in plasma 5 min after oral administration, and its concentration reached 2.47 μg/mL. Data from concentration-time curves allowed us to establish the main pharmacokinetic parameters in two models: one- and/or two-compartment. Cmax values were 3.33 and 3.42 μg/mL respectively, likewise AUC0→1440 min were 1,951.58 and 1,824.95 μg/mL·min. The G–A in brain tissue was noted to cross the blood-brain barrier, reaching Cmax 2.74 μg/mL, Tmax 81.6 min, and then drop gradually to 0.32 μg/mL detected at 24 h. The presence of G–A in brain tissue, confirmed that this anxiolytic compound can access the target organ and acts directly on the CNS.
Development of a Survey to Assess the Acceptability of an Innovative Contraception Practice among Rural Pharmacists
Authors: Wong Michael , Soon Judith , Zed Peter , Norman Wendy
Published: March 2014

Improved access to effective contraceptive methods is needed in Canada, particularly in rural areas, where unintended pregnancy rates are high and specific sexual health services may be further away. A rural pharmacist may be the most accessible health care professional. Pharmacy practice increasingly incorporates cognitive services. In Canada many provinces allow pharmacists to independently prescribe for some indications, but not for hormonal contraception. To assess the acceptability for the implementation of this innovative practice in Canada, we developed and piloted a survey instrument. We chose questions to address the components for adoption and change described in Rogers’ “diffusion of innovations” theory. The proposed instrument was iteratively reviewed by 12 experts, then focus group tested among eight pharmacists or students to improve the instrument for face validity, readability, consistency and relevancy to community pharmacists in the Canadian context. We then pilot tested the survey among urban and rural pharmacies. 4% of urban and 35% of rural pharmacies returned pilot surveys. Internal consistency on repeated re-phrased questions was high (Cronbach’s Alpha = 0.901). We present our process for the development of a survey instrument to assess the acceptability and feasibility among Canadian community pharmacists for the innovative practice of the independent prescribing of hormonal contraception.
Start a Research on Biopolymer Polyhydroxyalkanoate (PHA): A Review
Authors: Tan Giin-Yu , Chen Chia-Lung , Li Ling , Ge Liya , Wang Lin , Razaad Indah , Li Yanhong , Zhao Lei , Mo Yu , Wang Jing-Yuan
Published: March 2014

With the impending fossil fuel crisis, the search for and development of alternative chemical/material substitutes is pivotal in reducing mankind’s dependency on fossil resources. One of the potential substitute candidates is polyhydroxyalkanoate (PHA). PHA is a carbon-neutral and valuable polymer that could be produced from many renewable carbon sources by microorganisms, making it a sustainable and environmental-friendly material. At present, PHA is not cost competitive compared to fossil-derived products. Encouraging and intensifying research work on PHA is anticipated to enhance its economic viability in the future. The development of various biomolecular and chemical techniques for PHA analysis has led to the identification of many PHA-producing microbial strains, some of which are deposited in culture collections. Research work on PHA could be rapidly initiated with these ready-to-use techniques and microbial strains. This review aims to facilitate the start-up of PHA research by providing a summary of commercially available PHA-accumulating microbial cultures, PHA biosynthetic pathways, and methods for PHA detection, extraction and analysis.
Molecular Modeling of PEGylated Peptides, Dendrimers, and Single-Walled Carbon Nanotubes for Biomedical Applications
Authors: Lee Hwankyu
Published: March 2014

Polyethylene glycol (PEG) has been conjugated to many drugs or drug carriers to increase their solubility and circulating lifetime, and reduce toxicity. This has motivated many experimental studies to understand the effect of PEGylation on delivery efficiency. To complement the experimental findings and uncover the mechanism that cannot be captured by experiments, all-atom and coarse-grained molecular dynamics (MD) simulations have been performed. This has become possible, due to recent advances in simulation methodologies and computational power. Simulations of PEGylated peptides show that PEG chains wrap antimicrobial peptides and weaken their binding interactions with lipid bilayers. PEGylation also influences the helical stability and tertiary structure of coiled-coil peptides. PEGylated dendrimers and single-walled carbon nanotubes (SWNTs) were simulated, showing that the PEG size and grafting density significantly modulate the conformation and structure of the PEGylated complex, the interparticle aggregation, and the interaction with lipid bilayers. In particular, simulations predicted the structural transition between the dense core and dense shell of PEGylated dendrimers, the phase behavior of self-assembled complexes of lipids, PEGylated lipids, and SWNTs, which all favorably compared with experiments. Overall, these new findings indicate that simulations can now predict the experimentally observed structure and dynamics, as well as provide atomic-scale insights into the interactions of PEGylated complexes with other molecules.
Flotation of Biological Materials
Authors: Kyzas George , Matis Kostas
Published: March 2014

Flotation constitutes a gravity separation process, which originated from the minerals processing field. However, it has, nowadays, found several other applications, as for example in the wastewater treatment field. Concerning the necessary bubble generation method, typically dispersed-air or dissolved-air flotation was mainly used. Various types of biological materials were tested and floated efficiently, such as bacteria, fungi, yeasts, activated sludge, grape stalks, etc. Innovative processes have been studied in our Laboratory, particularly for metal ions removal, involving the initial abstraction of heavy metal ions onto a sorbent (including a biosorbent): in the first, the application of a flotation stage followed for the efficient downstream separation of metal-laden particles. The ability of microorganisms to remove metal ions from dilute aqueous solutions (as most wastewaters are) is a well-known property. The second separation process, also applied effectively, was a new hybrid cell of microfiltration combined with flotation. Sustainability in this field and its significance for the chemical and process industry is commented.
Multi-Mission Cross-Calibration of Satellite Altimeters: Constructing a Long-Term Data Record for Global and Regional Sea Level Change Studies
Authors: Bosch Wolfgang , Dettmering Denise , Schwatke Christian
Published: March 2014

Climate studies require long data records extending the lifetime of a single remote sensing satellite mission. Precise satellite altimetry exploring global and regional evolution of the sea level has now completed a two decade data record. A consistent long-term data record has to be constructed from a sequence of different, partly overlapping altimeter systems which have to be carefully cross-calibrated. This cross-calibration is realized globally by adjusting an extremely large set of single- and dual-satellite crossover differences performed between all contemporaneous altimeter systems. The total set of crossover differences creates a highly redundant network and enables a robust estimate of radial errors with a dense and rather complete sampling for all altimeter systems analyzed. An iterative variance component estimation is applied to obtain an objective relative weighting between altimeter systems with different performance. The final time series of radial errors is taken to estimate (for each of the altimeter systems) an empirical auto-covariance function. Moreover, the radial errors capture relative range biases and indicate systematic variations in the geo-centering of altimeter satellite orbits. The procedure has the potential to estimate for all altimeter systems the geographically correlated mean errors which is not at all visible in single-satellite crossover differences but maps directly to estimates of the mean sea surface.
Subpixelic Measurement of Large 1D Displacements: Principle, Processing Algorithms, Performances and Software
Authors: Guelpa Valérian , Laurent Guillaume , Sandoz Patrick , Zea July , Clévy Cédric
Published: March 2014

This paper presents a visual measurement method able to sense 1D rigid body displacements with very high resolutions, large ranges and high processing rates. Sub-pixelic resolution is obtained thanks to a structured pattern placed on the target. The pattern is made of twin periodic grids with slightly different periods. The periodic frames are suited for Fourier-like phase calculations—leading to high resolution—while the period difference allows the removal of phase ambiguity and thus a high range-to-resolution ratio. The paper presents the measurement principle as well as the processing algorithms (source files are provided as supplementary materials). The theoretical and experimental performances are also discussed. The processing time is around 3 µs for a line of 780 pixels, which means that the measurement rate is mostly limited by the image acquisition frame rate. A 3-σ repeatability of 5 nm is experimentally demonstrated which has to be compared with the 168 µm measurement range.
A Computational Model for Path Loss in Wireless Sensor Networks in Orchard Environments
Authors: Anastassiu Hristos , Vougioukas Stavros , Fronimos Theodoros , Regen Christian , Petrou Loukas , Zude Manuela , Käthner Jana
Published: March 2014

A computational model for radio wave propagation through tree orchards is presented. Trees are modeled as collections of branches, geometrically approximated by cylinders, whose dimensions are determined on the basis of measurements in a cherry orchard. Tree canopies are modeled as dielectric spheres of appropriate size. A single row of trees was modeled by creating copies of a representative tree model positioned on top of a rectangular, lossy dielectric slab that simulated the ground. The complete scattering model, including soil and trees, enhanced by periodicity conditions corresponding to the array, was characterized via a commercial computational software tool for simulating the wave propagation by means of the Finite Element Method. The attenuation of the simulated signal was compared to measurements taken in the cherry orchard, using two ZigBee receiver-transmitter modules. Near the top of the tree canopies (at 3 m), the predicted attenuation was close to the measured one—just slightly underestimated. However, at 1.5 m the solver underestimated the measured attenuation significantly, especially when leaves were present and, as distances grew longer. This suggests that the effects of scattering from neighboring tree rows need to be incorporated into the model. However, complex geometries result in ill conditioned linear systems that affect the solver’s convergence.
A New Direct Single-Molecule Observation Method for DNA Synthesis Reaction Using Fluorescent Replication Protein A
Authors: Takahashi Shunsuke , Kawasaki Shohei , Miyata Hidefumi , Kurita Hirofumi , Mizuno Takeshi , Matsuura Shun-ichi , Mizuno Akira , Oshige Masahiko , Katsura Shinji
Published: March 2014

Using a single-stranded region tracing system, single-molecule DNA synthesis reactions were directly observed in microflow channels. The direct single-molecule observations of DNA synthesis were labeled with a fusion protein consisting of the ssDNA-binding domain of a 70-kDa subunit of replication protein A and enhanced yellow fluorescent protein (RPA-YFP). Our method was suitable for measurement of DNA synthesis reaction rates with control of the ssλDNA form as stretched ssλDNA (+flow) and random coiled ssλDNA (−flow) via buffer flow. Sequentially captured photographs demonstrated that the synthesized region of an ssλDNA molecule monotonously increased with the reaction time. The DNA synthesis reaction rate of random coiled ssλDNA (−flow) was nearly the same as that measured in a previous ensemble molecule experiment (52 vs. 50 bases/s). This suggested that the random coiled form of DNA (−flow) reflected the DNA form in the bulk experiment in the case of DNA synthesis reactions. In addition, the DNA synthesis reaction rate of stretched ssλDNA (+flow) was approximately 75% higher than that of random coiled ssλDNA (−flow) (91 vs. 52 bases/s). The DNA synthesis reaction rate of the Klenow fragment (3’-5’exo–) was promoted by DNA stretching with buffer flow.
Design and Experimentation with Sandwich Microstructure for Catalytic Combustion-Type Gas Sensors
Authors: Gu Jun-Tao , Zhang Yong-De , Jiang Jin-Gang
Published: March 2014

The traditional handmade catalytic combustion gas sensor has some problems such as a pairing difficulty, poor consistency, high power consumption, and not being interchangeable. To address these issues, integrated double catalytic combustion of alcohol gas sensor was designed and manufactured using silicon micro-electro-mechanical systems (MEMS) technology. The temperature field of the sensor is analyzed using the ANSYS finite element analysis method. In this work, the silicon oxide-PECVD-oxidation technique is used to manufacture a SiO2-Si3N2-SiO2 microstructure carrier with a sandwich structure, while wet etching silicon is used to form a beam structure to reduce the heat consumption. Thin-film technology is adopted to manufacture the platinum-film sensitive resistance. Nano Al2O3-ZrO-ThO is coated to format the sensor carrier, and the sensitive unit is dipped in a Pt-Pd catalyst solution to form the catalytic sensitive bridge arm. Meanwhile the uncoated catalyst carrier is considered as the reference unit, realizing an integrated chip based on a micro double bridge and forming sensors. The lines of the Pt thin-film resistance have been observed with an electronic microscope. The compensation of the sensitive material carriers and compensation materials have been analyzed using an energy spectrum. The results show that the alcohol sensor can detect a volume fraction between 0 and 4,500 × 10−6 and has good linear output characteristic. The temperature ranges from −20 to +40 °C. The humidity ranges from 30% to 85% RH. The zero output of the sensor is less than ±2.0% FS. The power consumption is ≤0.2 W, and both the response and recovery time are approximately 20 s.
Force Sensor Attachable to Thin Fiberscopes/Endoscopes Utilizing High Elasticity Fabric
Authors: Watanabe Tetsuyou , Iwai Takanobu , Fujihira Yoshinori , Wakako Lina , Kagawa Hiroyuki , Yoneyama Takeshi
Published: March 2014

An endoscope/fiberscope is a minimally invasive tool used for directly observing tissues in areas deep inside the human body where access is limited. However, this tool only yields visual information. If force feedback information were also available, endoscope/fiberscope operators would be able to detect indurated areas that are visually hard to recognize. Furthermore, obtaining such feedback information from tissues in areas where collecting visual information is a challenge would be highly useful. The major obstacle is that such force information is difficult to acquire. This paper presents a novel force sensing system that can be attached to a very thin fiberscope/endoscope. To ensure a small size, high resolution, easy sterilization, and low cost, the proposed force visualization–based system uses a highly elastic material—panty stocking fabric. The paper also presents the methodology for deriving the force value from the captured image. The system has a resolution of less than 0.01 N and sensitivity of greater than 600 pixels/N within the force range of 0–0.2 N.
Olfaction-Inspired Sensing Using a Sensor System with Molecular Recognition and Optimal Classification Ability for Comprehensive Detection of Gases
Authors: Imahashi Masahiro , Watanabe Masashi , Jha Sunil , Hayashi Kenshi
Published: March 2014

In this study, we examined the comprehensive detection of numerous volatile molecules based on the olfactory information constructed by using olfaction-inspired sensor technology. The sensor system can simultaneously detect multiple odors by the separation and condensation ability of molecularly imprinted filtering adsorbents (MIFAs), where a MIP filter with a molecular sieve was deposited on a polydimethylsiloxane (PDMS) substrate. The adsorption properties of MIFAs were evaluated using the solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS). The results demonstrated that the system embedded with MIFAs possesses high sensitivity and specific selectivity. The digitization and comprehensive classification of odors were accomplished by using artificial odor maps constructed through this system.
A Holistic Approach to Enhance the Use of Neglected and Underutilized Species: The Case of Andean Grains in Bolivia and Peru
Authors: Padulosi Stefano , Amaya Karen , Jäger Matthias , Gotor Elisabetta , Rojas Wilfredo , Valdivia Roberto
Published: March 2014

The IFAD-NUS project, implemented over the course of a decade in two phases, represents the first UN-supported global effort on neglected and underutilized species (NUS). This initiative, deployed and tested a holistic and innovative value chain framework using multi-stakeholder, participatory, inter-disciplinary, pro-poor gender- and nutrition-sensitive approaches. The project has been linking aspects often dealt with separately by R&D, such as genetic diversity, selection, cultivation, harvest, value addition, marketing, and final use, with the goal to contribute to conservation, better incomes, and improved nutrition and strengthened livelihood resilience. The project contributed to the greater conservation of Andean grains and their associated indigenous knowledge, through promoting wider use of their diversity by value chain actors, adoption of best cultivation practices, development of improved varieties, dissemination of high quality seed, and capacity development. Reduced drudgery in harvest and postharvest operations, and increased food safety were achieved through technological innovations. Development of innovative food products and inclusion of Andean grains in school meal programs is projected to have had a positive nutrition outcome for targeted communities. Increased income was recorded for all value chain actors, along with strengthened networking skills and self-reliance in marketing. The holistic approach taken in this study is advocated as an effective strategy to enhance the use of other neglected and underutilized species for conservation and livelihood benefits.
In-vitro Neurotoxicity of Two Malaysian Krait Species (Bungarus candidus and Bungarus fasciatus) Venoms: Neutralization by Monovalent and Polyvalent Antivenoms from Thailand
Authors: Rusmili Muhamad , Yee Tee , Mustafa Mohd , Othman Iekhsan , Hodgson Wayne
Published: March 2014

Bungarus candidus and Bungarus fasciatus are two species of krait found in Southeast Asia. Envenoming by these snakes is often characterized by neurotoxicity and, without treatment, causes considerable morbidity and mortality. In this study, the in vitro neurotoxicity of each species, and the effectiveness of two monovalent antivenoms and a polyvalent antivenom, against the neurotoxic effects of the venoms, were examined in a skeletal muscle preparation. Both venoms caused concentration-dependent inhibition of indirect twitches, and attenuated responses to exogenous nicotinic receptor agonists, in the chick biventer preparation, with B. candidus venom being more potent than B. fasciatus venom. SDS-PAGE and western blot analysis indicated different profiles between the venoms. Despite these differences, most proteins bands were recognized by all three antivenoms. Antivenom, added prior to the venoms, attenuated the neurotoxic effect of the venoms. Interestingly, the respective monovalent antivenoms did not neutralize the effects of the venom from the other Bungarus species indicating a relative absence of cross-neutralization. Addition of a high concentration of polyvalent antivenom, at the t90 time point after addition of venom, partially reversed the neurotoxicity of B. fasciatus venom but not B. candidus venom. The monovalent antivenoms had no significant effect when added at the t90 time point. This study showed that B. candidus and B. fasciatus venoms display marked in vitro neurotoxicity in the chick biventer preparation and administration of antivenoms at high dose is necessary to prevent or reverse neurotoxicity.
Viral Metagenomics: Analysis of Begomoviruses by Illumina High-Throughput Sequencing
Authors: Idris Ali , Al-Saleh Mohammed , Piatek Marek , Al-Shahwan Ibrahim , Ali Shahjahan , Brown Judith
Published: March 2014

Traditional DNA sequencing methods are inefficient, lack the ability to discern the least abundant viral sequences, and ineffective for determining the extent of variability in viral populations. Here, populations of single-stranded DNA plant begomoviral genomes and their associated beta- and alpha-satellite molecules (virus-satellite complexes) (genus, Begomovirus; family, Geminiviridae) were enriched from total nucleic acids isolated from symptomatic, field-infected plants, using rolling circle amplification (RCA). Enriched virus-satellite complexes were subjected to Illumina-Next Generation Sequencing (NGS). CASAVA and SeqMan NGen programs were implemented, respectively, for quality control and for de novo and reference-guided contig assembly of viral-satellite sequences. The authenticity of the begomoviral sequences, and the reproducibility of the Illumina-NGS approach for begomoviral deep sequencing projects, were validated by comparing NGS results with those obtained using traditional molecular cloning and Sanger sequencing of viral components and satellite DNAs, also enriched by RCA or amplified by polymerase chain reaction. As the use of NGS approaches, together with advances in software development, make possible deep sequence coverage at a lower cost; the approach described herein will streamline the exploration of begomovirus diversity and population structure from naturally infected plants, irrespective of viral abundance. This is the first report of the implementation of Illumina-NGS to explore the diversity and identify begomoviral-satellite SNPs directly from plants naturally-infected with begomoviruses under field conditions.
Management of Tundra Wastewater Treatment Wetlands within a Lagoon/Wetland Hybridized Treatment System Using the SubWet 2.0 Wetland Model
Authors: Chouinard Annie , Yates Colin , Balch Gordon , Jørgensen Sven , Wootton Brent , Anderson Bruce
Published: March 2014

The benefits provided by natural (e.g., non-engineered) tundra wetlands for the treatment of municipal wastewater in the Canadian Arctic are largely under-studied and, therefore, undervalued in regard to the treatment service wetlands provide to small remote Arctic communities. In this paper we present case studies on two natural tundra systems which at the time of study had different management practices, in which one consisted of a facultative lake system continuously discharging into a tundra wetland, while the second system had wastewater discharged directly into a tundra wetland. We also examine the utility of the SubWet 2.0 wetland model and how it can be used to: (i) predict the outcomes of management options; and (ii) to assess treatment capacity within individual tundra wetlands to meet future needs associated with population growth and to help municipalities determine the appropriate actions required to achieve the desired level of treatment, both currently, and in a sustainable long-term manner. From this examination we argue that tundra wetlands can significantly augment common treatment practices which rely on waste stabilization ponds, by recognizing the services that wetlands already provide. We suggest that treatment targets could be more achievable if tundra wetlands are formally recognized as part of a hybridized treatment system that incorporates the combined benefits of both the waste stabilization pond and the tundra wetland. Under this scenario tundra wetlands would be recognized as part of the treatment process and not as the ‘receiving’ environment, which is how most tundra wetlands are currently categorized.
TIM4 Regulates the Anti-Islet Th2 Alloimmune Response
Authors: Vergani Andrea , Gatti Francesca , Lee Kang M. , D'addio Francesca , Tezza Sara , Chin Melissa , Bassi Roberto , Tian Ze , Wu Erxi , Maffi Paola , Ben Nasr Moufida , Kim James I. , Secchi Antonio , Markmann James F. , Rothstein David M. , Turka Laurence A. , Sayegh Mohamed H. , Fiorina Paolo
Published: March 2014

The role of the novel costimulatory molecule TIM4 in anti-islet response is unknown. We explored TIM4 expression and targeting in Th1 (BALB/c islets into C57BL/6 mice) and Th2 (BALB/c islets into Tbet-/- C57BL/6 mice) models of anti-islet alloimmune response and in a model of anti-islet autoimmune response (diabetes onset in NOD mice). The targeting of TIM4, using the monoclonal antibody RMT4-53, promotes islet graft survival in a Th1 model, with 30% of the graft surviving in the long term; islet graft protection appears to be mediated by a Th1 to Th2 skewing of the immune response. Differently, in the Th2 model, TIM4 targeting precipitates graft rejection by further enhancing the Th2 response. The effect of anti-TIM4 treatment in preventing autoimmune diabetes was marginal with only minor Th1 to Th2 skewing. B-Cell depletion abolished the effect of TIM4 targeting. TIM4 is expressed on human B-cells and is upregulated in diabetic and islet-transplanted patients. Our data suggest a model in which TIM4 targeting promotes Th2 response over Th1 via B-cells. The targeting of TIM4 could become a component of an immunoregulatory protocol in clinical islet transplantation, aiming at redirecting the immune system toward a Th2 response.
Mesenchymal Stem Cells Increase Hippocampal Neurogenesis and Neuronal Differentiation by Enhancing the Wnt Signaling Pathway in an Alzheimer's Disease Model
Authors: Oh Se Hee , Kim Ha Na , Park Hyun-Jung , Shin Jin Young , Lee Phil Hyu
Published: March 2014

Neurogenesis in the subgranular zone of the hippocampal dentate gyrus may act as an endogenous repair mechanism in Alzheimer's disease (AD), and the Wnt signaling pathway has been suggested to closely modulate neurogenesis in amyloid-β (Aβ)-related AD models. The present study investigated whether mesenchymal stem cells (MSCs) would modulate hippocampal neurogenesis via modulation of the Wnt signaling pathway in a model of AD. In Aβ-treated neuronal progenitor cells (NPCs), the coculture with MSCs increased significantly the expression of Ki-67, GFAP, SOX2, nestin, and HuD compared to Aβ treatment alone. In addition, MSC treatment in Aβ-treated NPCs enhanced the expression of β-catenin and Ngn1 compared to Aβ treatment alone. MSC treatment in Aβ-treated animals significantly increased the number of BrdU-ir cells in the hippocampus at 2 and 4 weeks compared to Aβ treatment alone. In addition, quantitative analysis showed that the number of BrdU and HuD double-positive cells in the dentate gyrus was significantly higher in the MSC-treated group than in controls or after Aβ treatment alone. These results demonstrate that MSC administration significantly augments hippocampal neurogenesis and enhances the differentiation of NPCs into mature neurons in AD models by augmenting the Wnt signaling pathway. The use of MSCs to modulate endogenous adult neurogenesis may have a significant impact on future strategies for AD treatment.
Frequency, amplitude, and phase measurements in contact resonance atomic force microscopies
Authors: Stan Gheorghe , Solares Santiago D
Published: March 2014

The resonance frequency, amplitude, and phase response of the first two eigenmodes of two contact-resonance atomic force microscopy (CR-AFM) configurations, which differ in the method used to excite the system (cantilever base vs sample excitation), are analyzed in this work. Similarities and differences in the observables of the cantilever dynamics, as well as the different effect of the tip–sample contact properties on those observables in each configuration are discussed. Finally, the expected accuracy of CR-AFM using phase-locked loop detection is investigated and quantification of the typical errors incurred during measurements is provided.
STATISTICAL SURVEY OF THE PATHOLOGY, DIAGNOSIS AND REHABILITATION OF ETICS IN WALLS
Authors: Amaro Bárbara , Saraiva Diogo , Brito Jorge de , Flores-Colen Inês
Published: March 2014

This paper presents a statistical survey on the pathology, diagnosis and rehabilitation of ETICS (external thermal insulation composite system) in walls. The development of an expert-knowledge inspection and diagnosis system for this type of cladding implied the creation of classification lists of anomalies, most probable causes, diagnosis methods and repair/maintenance techniques. The collection of data through field work where the tools developed within the proposed system were applied was the basis for this statistical survey. The field work consisted of the visual inspection of 146 façades with ETICS cladding located in various areas in Portugal and aged from 3 to 22 years. The data obtained enable anomalies to be related to their causes and so conclusions can be drawn on the best way to minimize the development of the former and eliminate the latter, at the design and application stages. The study further revealed the main sensitive issues of the cladding system during its service life, highlighting the importance of a correct maintenance plan.
Reduced Toll-Like Receptor 9 Expression on Peripheral C D14+ Monocytes of Chronic Hepatitis B Patients and its Restoration by Effective Therapy
Authors: Huang Yi-Wen , Hsu Cheng-Kai , Lin Shih-Chang , Wei Shu-Chen , Hu Jui-Ting , Chang Han-Yu , Liang Cher-Wei , Chen Ding-Shinn , Chen Pei-Jer , Hsu Ping-Ning , Yang Sien-Sing , Kao Jia-Horng
Published: March 2014

Background

Chronic hepatitis B (CHB) patients display Toll-like receptor 9 (TLR9)-dependent defective immune responses. We aimed to study TLR9 expression on CHB patients and its alteration during therapy.

Methods

We compared TLR9 expression on fresh peripheral CD14+ monocytes from a cohort of 97 CHB patients and 35 HBsAg-negative, anti-HCV-negative controls, during pegylated interferon or entecavir therapy. TLR9 expression on liver tissue was also investigated.

Results

Compared with controls, peripheral CD14+ monocytes of CHB patients displayed reduced expression of TLR9 mean fluorescence intensity (MFI; 9.90 ±3.64 versus 7.95 ±3.61; P=0.007) independent of age, gender and alanine aminotransferase (ALT; -2.09, 95% CI -3.568, -0.613; P=0.006). Furthermore, age, gender, ALT, HBeAg status, quantitative HBsAg (qHBsAg) or HBV DNA did not predict the TLR9 expression ( P=0.863). Hepatic TLR9 messenger RNA (mRNA) was significantly reduced in 54 patients compared with 3 controls (0.45 ±0.32 versus 1-fold). Using response-guided therapy by qHBsAg levels and pretreatment TLR9 MFI as a reference, TLR9 MFI restored to a mean of 1.7- to 2.7-fold in pegylated interferon responders and reduced to a mean of 0.6-to 0.7-fold in non-responders starting from treatment week 12. Among 10 entecavir-treated patients, TLR9 MFI gradually restored to a mean of 1.2- to 2.1-fold starting from treatment week 48.

Conclusions

CHB patients display reduced TLR9 expression on peripheral CD14+ monocytes, which is independent of host and viral markers, and on liver tissue. Responders to pegylated interferon and those under entecavir demonstrate restoration of TLR9 expression. On-treatment TLR9 expression on peripheral monocytes might predict response to pegylated interferon therapy.
Prevalence of Baseline Polymorphisms for Potential Resistance to NS5A Inhibitors in Drug-Naive Individuals Infected with Hepatitis C Genotypes 1–4
Authors: McCormick Adele L , Wang Lawrence , Garcia-Diaz Ana , Macartney Malcolm J , Webster Daniel P , Haque Tanzina
Published: March 2014

Background

The non-structural 5A (NS5A) protein of HCV is a multifunctional phosphoprotein involved in regulation of viral replication and virion assembly. NS5A inhibitors targeting domain I of NS5A protein have demonstrated high potency and pan-genotypic antiviral activity, however they possess a low genetic barrier to resistance. At present, only genotype 1, the most prevalent HCV genotype has been studied in detail for resistant variants.

Methods

Utilizing a panel of genotypic-specific resistance assays, population sequencing was performed on plasma-derived viral RNA isolated from 138 patients infected with HCV genotypes 1–4 and not treated with direct-acting antiviral agents. Amino acid changes in HCV NS5A domain I at codon positions 28, 30, 31, 32 and 93, reported to confer reduced susceptibility to certain NS5A inhibitors were examined. Additionally, genotypic outcome based on NS5A sequences were compared with VERSANT HCV Genotype Assay (LiPA) 1.0 (Siemens Healthcare Diagnostics, Surrey, UK) and Abbott m2000 RealTi me HCV genotype II assay (Abbott Molecular, Maidenhead, Berkshire, UK).

Results

Amino acid substitutions associated with moderate to high level resistance to NS5A inhibitors were detected in 2/42 (4.76%) HCV-1a, 3/23 (13.04%) HCV-1b, 4/26 (15.38%) HCV-2, 1/24 (4.17%) HCV-3 and 1/23 (4.35%) HCV-4 infected patients who had not been treated with NS5A inhibitors. Genotype prediction based on NS5A sequences were concordant with LiPA and/or Abbott Real-Ti me for 97.10% of cases.

Conclusions

Primary resistance mutations associated with resistance to first-generation NS5A inhibitors such as daclatasvir were observed in all genotypes, albeit at low frequencies. An excellent correlation based on NS5A genotyping and LiPA or Abbott Real-Ti me was achieved.
Sodium Tanshinone IIA Sulfonate Inhibits the MEQ, Ul49 and VP22 Expression of Marek's Disease Virus
Authors: Sun Na , Cong Xiang-Ming , Jiang Jun-Bing , Zhao Jun-Xing , Wang Wen-Kui , Duan Zhi-Bian , Hu Yuan-Liang , Lei Hai-Min , Li Hong-Quan
Published: March 2014

Background

Our previous studies have demonstrated that sodium tanshinone IIA sulfonate (STS), a natural compound derived from Salviae Miltiorrhizae Radix et Rhizoma (Danshen), could effectively inhibit Marek's disease virus (MDV) infection both in vitro and in vivo, but the underlying mechanisms remain unclear. The main objective of the study was to explore the effect of STS on the meq, ul49 and VP22 expression of MDV in vitro.

Methods

Quantitative real-time PCR (qRT-PCR) was used to analyse the effect of STS on meq and ul49 expression at both the DNA and messenger RNA (mRNA) level, and the effect of STS on VP22 was assessed by immunofluorescence assay and western blotting.

Results

The DNA and mRNA copy numbers of meq and ul49 significantly decreased in the groups treated with STS compared with MDV control ( P<0.05), which indicated that STS could inhibit the expression of meq and ul49 at both the DNA and mRNA level. Moreover, the expression of VP22 encoded by ul49 was also significantly inhibited ( P<0.05).

Conclusions

STS possessed anti-MDV activity in chicken embryo fibroblasts. Its antiviral mechanisms may be ascribed to inactivating MDV directly, disturbing meq and ul49 replication and inhibiting the expression of VP22 encoded by ul49. These results suggested that STS is a promising natural compound to be further developed as an antiviral agent against MDV infection.
Molecular epidemiology and virulence characteristics of prevalent group A streptococci recovered from patients in northern India
Authors: Arya Deepak Kumar , Sharma Abhinay , Mehta Geeta , Dua Meenakshi , Johri Atul Kumar
Published: March 2014

Introduction: In this study, the prevalence of M types of Group A Streptococcus (GAS) in North India, invasive behavior of prevalent M types, and inflammatory immune response by host cells were studied. Methodology: A total of 1,047 clinical samples were collected between 2004 and 2010. Confirmation of GAS was determined by serotyping and M types were identified by emm gene sequencing. The most prevalent serotypes were selected to study their invasive behavior and inflammatory immune response under different temperatures and salt concentrations in A549 and HEp-2 cells. Results: Ninety-two isolates were identified as GAS of which 17 were M types with 18.5% heterogeneity. The most prevalent M types were M1 (21.73%) and M49 (8.7%), respectively. M1 and M49 were used to study virulence potential and inflammatory immune responses. The efficiency of cell infection decreased with increased temperature for both M types, increasing with lowering temperatures compared to the uninfected control (37°C). As salt concentration was increased, cell infection efficiency was lowered with some exceptions; the infection efficiency of M1 strain in A549 cells with 0.6 M NaCl was 50 fold higher (p ≤ 0.03). Significantly increased production of IL-6 and IL-8 was observed in both cell lines infected with GAS and when grown under different environmental conditions compared to uninfected cell lines. Conclusions: This study determined the prevalence of different M types in North India and showed that environmental conditions can regulate cell infection by GAS . This information may influence the selection of GAS serotypes used in vaccine development.