bluuebunny/crossref_metadata_2025 huggingface dataset:
Vibration-Induced Pressures on a Cylindrical Structure Surface in Compressible Fluid
Authors: Liu Ping , Tang Bai-Jian , Kaewunruen Sakdirat
Published: April 2019

This paper unprecedentedly addresses the effect of vibrations of a cylindrical structure on dynamic pressures in a compressible and incompressible fluid situation. To obtain analytical solutions, the density of the fluid is simplified as a constant, but the rates of the density with respect to time and to space are considered as a dynamic and time-dependent function. In addition, the low velocity of the vibration is taken into account so the lower order terms are negligible. According to the assumption that the vibration at the boundary of the structure behaves as a harmonic function, some interesting and new analytical solutions can be established. Both analytical solutions in the cases of the compressible and incompressible fluid are rigorously verified by the calibrated numerical simulations. New findings reveal that, in the case of the incompressible fluid, dynamic pressure at the surface of the cylindrical shell is proportional to the acceleration of the vibration, which acts like an added mass. In the case of the compressible fluid, the pressure at the surface of the cylindrical structure is proportional to the velocity of the vibration, which acts as a damping. In addition, the proportional ratio is derived as ρ c .
Fertility Detection of Hatching Eggs Based on a Convolutional Neural Network
Authors: Geng Lei , Hu Yuzhou , Xiao Zhitao , Xi Jiangtao
Published: April 2019

In order to achieve the goal of detecting the fertility of hatching eggs which are divided into fertile eggs and dead eggs more accurately and effectively, a novel method combining a convolution neural network (CNN) and a heartbeat signal of the hatching eggs is proposed in this paper. Firstly, we collected heartbeat signals of 9-day-later hatching eggs by the method of PhotoPlethysmoGraphy (PPG), which is a non-invasive method to detect the change of blood volume in living tissues by photoelectric means. Secondly, a sequential convolutional neural network E-CNN, which was used to analyze heartbeat sequence of hatching eggs, was designed. Thirdly, an end-to-end trainable convolutional neural network SR-CNN, which was used to process heartbeat waveform images of hatching eggs, was designed to improve the classification performance in this paper. Key to improving the classification performance of SR-CNN is the SE-Res module, which combines the channel weighting unit “Squeeze-and-Excitation” (SE) block and the residual structure. The experimental results show that two models trained on our dataset, with E-CNN and SR-CNN, are able to achieve the fertility detection of the hatching eggs with superior identification accuarcy, up to 99.50% and 99.62% respectively, on our test set. It is demonstrated that the proposed method is feasible for identifying and classifying the survival of hatching eggs accurately and effectively.
Influence of Numerical Aperture on Molten Area Formation in Fusion Micro-Welding of Glass by Picosecond Pulsed Laser
Authors: Ouyang Zhiyong , Okamoto Yasuhiro , Ogino Yuta , Sakagawa Tomokazu , Okada Akira
Published: April 2019

Focusing condition such as numerical aperture (N.A.) has a great influence on the creation of molten area and the stable welding process in fusion micro-welding of glass. In this study, a picosecond pulsed laser of 1064 nm in wavelength and 12.5 ps in pulse duration was tightly focused inside a borosilicate glass using objective lenses of numerical apertures 0.45, 0.65, and 0.85 with spherical aberration correction. Influence of numerical aperture on molten area formation was experimentally investigated through analysis of focusing situation in glass, and movement of absorption point, and then molten area characteristics were discussed. It is concluded that N.A. of 0.65 with superior focusing characteristics can form a large and continuous molten area without cracks, which enables achievement of stable joining of glass material by picosecond pulsed laser.
Identifying Key Sources of City Air Quality: A Hybrid MCDM Model and Improvement Strategies
Authors: Chen Kou-Hsiung , Yien Jui-Mei , Chiang Cheng-Hsin , Tsai Pei-Chun , Tsai Fu-Sheng
Published: April 2019

Improvements for air quality were prioritized according to gaps between criteria. Existing improvement strategies for air quality often focus on a single criterion, without considering associations among criteria. Moreover, solutions are often temporary, failing to provide long-term improvement. Therefore, this study employed a multiple-criteria decision-making model and a substitution method based on causal relationships to analyze potential improvement strategies for air quality in Kaohsiung, Taiwan. Results revealed that coal-fired power plants and factory emissions are the major sources of pollution in Kaohsiung. This study suggests that environmental authorities in Kaohsiung should facilitate plants to improve energy efficiency with anti-pollution facilities.
Effect of Soil Reinforcement on Tunnel Deformation as a Result of Stress Relief
Authors: Sun Huasheng , Sun Wenbin
Published: April 2019

Adjacent geotechnical engineering activities, such as deep excavation, may adversely affect or even damage adjacent tunnels. Ground reinforcement before excavation may be an effective approach to reduce tunnel heave as a result of stress relief. However, there are few quantitative studies on the effect of soil reinforcement on tunnel deformation. Moreover, the reinforcement mechanism of the reinforced soil and the reinforcement depth are not fully understood. In order to investigate the effect of reinforcing the ground on the tunnel response, a finite element analysis was conducted based on a previously reported centrifugal model test with no ground reinforcement. The effect of the Young’s modulus and depth of the reinforced soil on tunnel deformation was analyzed. Soil stresses around the tunnel were also considered to explain the tunnel response. The results revealed that the Young’s modulus of the reinforced soil and the reinforcement depth had a significant impact on tunnel deformation as a result of basement excavation. The tunnel heave in the longitudinal direction decreased by 18% and 27% for modulus of the reinforced soil, five times and ten times higher than that of the non-reinforced soil, respectively. The reinforcement depth was effective with regard to controlling the tunnel heave caused by stress relief. This is because the reinforced soil blocked the stress transfer and thus reduced the tunnel heave caused by excavation unloading. It is expected that this study will be useful with regard to taking effective measures and ensuring the safety and serviceability of existing metro tunnels during adjacent excavation.
One-Dimensional Nanostructure Engineering of Conducting Polymers for Thermoelectric Applications
Authors: Shah Kwok Wei , Wang Su-Xi , Soo Debbie Xiang Yun , Xu Jianwei
Published: April 2019

The past few decades have witnessed considerable progress of conducting polymer-based organic thermoelectric materials due to their significant advantages over the traditional inorganic materials. The nanostructure engineering and performance investigation of these conducting polymers for thermoelectric applications have received considerable interest but have not been well documented. This review gives an outline of the synthesis of various one-dimensional (1D) structured conducting polymers as well as the strategies for hybridization with other nanomaterials or polymers. The thermoelectric performance enhancement of these materials in association with the unique morphologies and structures are discussed. Finally, perspectives and suggestions for the future research based on these interesting nanostructuring methodologies for improvement of thermoelectric materials are also presented.
Multi-Agent Planning under Uncertainty with Monte Carlo Q-Value Function
Authors: Zhang Jian , Pan Yaozong , Wang Ruili , Fang Yuqiang , Yang Haitao
Published: April 2019

Decentralized partially observable Markov decision processes (Dec-POMDPs) are general multi-agent models for planning under uncertainty, but are intractable to solve. Doubly exponential growth of the search space as the horizon increases makes a brute-force search impossible. Heuristic methods can guide the search towards the right direction quickly and have been successful in different domains. In this paper, we propose a new Q-value function representation—Monte Carlo Q-value function Q MC , which is proved to be an upper bound of the optimal Q-value function Q * . We introduce two Monte Carlo tree search enhancements—heavy playout for a simulation policy and adaptive samples—to speed up computation of Q MC . Then, we present a clustering and expansion with Monte-Carlo algorithm (CEMC)—an offline planning algorithm using Q MC as Q-value function, which is based on the generalized multi-agent A* with incremental clustering and expansion (GMAA*-ICE or ICE). CEMC calculates Q-value functions as required, without computing and storing all Q-value functions. An extended policy pruning strategy is used in CEMC. Finally, we present empirical results demonstrating that CEMC outperforms the best heuristic algorithm with a compact Q-value presentation in term of runtime for the same horizon, and has less memory usage for larger problems.
Understanding Spatial Variability of Air Quality in Sydney: Part 1—A Suburban Balcony Case Study
Authors: Simmons Jack B. , Paton-Walsh Clare , Phillips Frances , Naylor Travis , Guérette Élise-Andrée , Burden Sandy , Dominick Doreena , Forehead Hugh , Graham Joel , Keatley Thomas , Gunashanhar Gunaratnam , Kirkwood John
Published: April 2019

There is increasing awareness in Australia of the health impacts of poor air quality. A common public concern raised at a number of “roadshow” events as part of the federally funded Clean Air and Urban Landscapes Hub (CAUL) project was whether or not the air quality monitoring network around Sydney was sampling air representative of typical suburban settings. In order to investigate this concern, ambient air quality measurements were made on the roof of a two-storey building in the Sydney suburb of Auburn, to simulate a typical suburban balcony site. Measurements were also taken at a busy roadside and these are discussed in a companion paper (Part 2). Measurements made at the balcony site were compared to data from three proximate regulatory air quality monitoring stations: Chullora, Liverpool and Prospect. During the 16-month measurement campaign, observations of carbon monoxide, oxides of nitrogen, ozone and particulate matter less than 2.5-µm diameter at the simulated urban balcony site were comparable to those at the closest permanent air quality stations. Despite the Auburn site experiencing 10% higher average carbon monoxide amounts than any of the permanent air quality monitoring sites, the oxides of nitrogen were within the range of the permanent sites and the pollutants of greatest concern within Sydney (PM2.5 and ozone) were both lowest at Auburn. Similar diurnal and seasonal cycles were observed between all sites, suggesting common pollutant sources and mechanisms. Therefore, it is concluded that the existing air quality network provides a good representation of typical pollution levels at the Auburn “balcony” site.
Nitrogen Interstitial Alloying of CoCrFeMnNi High Entropy Alloy through Reactive Powder Milling
Authors: Moravcik Igor , Cizek Jan , Gouvea Larissa de Almeida , Cupera Jan , Guban Ivan , Dlouhy Ivo
Published: April 2019

The present work is focused on the synthesis of CoCrFeMnNi high entropy alloy (HEA) interstitially alloyed with nitrogen via powder metallurgy routes. Using a simple method, nitrogen was introduced to the HEA from the protective N2 gas atmosphere during mechanical alloying (MA) processing. The lattice parameter and amount of nitrogen in HEA were observed to be linearly proportional to the milling duration. The limited solubility of nitrogen in the main face centered cubic (FCC) phase resulted in the in-situ formation of nitrides and, accordingly, significant increase in the hardness values. It has been shown that fabrication of such nitrogen-doped HEA bulk materials can be conveniently achieved by a simple combination of MA + spark plasma sintering processes, without the need for adding nitrogen from other sources.
Observation Research on the Effect of UHVDC Grounding Current on Buried Pipelines
Authors: Liu Lianguang , Yu Zebang , Jiang Zhe , Hao Jianhong , Liu Wenlin
Published: April 2019

In order to research the electromagnetic interference in buried oil and gas pipelines generated by the grounding current of a grounding electrode of Ultra High Voltage Direct Current (UHVDC) system, observation experiments of stray current and pipe-to-soil potential (PSP) in the pipeline were carried out. Monitoring devices were installed at two sites of the Dong-Huang oil pipeline before the commissioning of the Zalute–Qingzhou and Shanghaimiao–Shandong ±800 kV UHVDC projects. Monitoring data on the stray current and PSP of the two monitoring sites were obtained when the two UHVDC projects were operated in monopolar mode on 24 December 2017 and 2 January 2019 and the grounding current reached 6250 A. The amplitude characteristics of the stray current and PSP at different distances from the grounding electrode and the effects of the magnitude of the stray current and PSP on the cathodic protection system are analyzed herein. The results show that the effects of the grounding current on pipeline corrosion are not only closely related to the distance between the grounding electrode and the pipeline but are also related to the running state of the potentiostats of pipelines and the distance between insulation flanges. Optimizing the performance of potentiostats and the distribution of insulation flanges can reduce the effects of UHVDC grounding current on pipeline corrosion.
Numerical Delineation of 3D Unsteady Flow Fields in Side Channel Pumps for Engineering Processes
Authors: Zhang Fan , Chen Ke , Appiah Desmond , Hu Bo , Yuan Shouqi , Asomani Stephen Ntiri
Published: April 2019

Side channel pumps are important machines for handling toxic, explosive or other dangerous liquids in various engineering processes. However, the operational reliability of these pumps is directly affected by the intensity of the pressure and velocity fluctuations, thus the flow fluctuations existing within the pump cannot be neglected because of their direct influence on the noise, vibration and harshness performance. Therefore, describing precisely the zones of highly unsteady and turbulent flow fields is a key research topic. Moreover, the size of the wrapping angle strongly affects the levels of pressure and velocity fluctuations, thus numerical calculations of the pressure and velocity fluctuation intensities in side channel pump models with different wrapping angles were conducted in this work. The results indicated that the pressure fluctuation coefficient increased gradually from the inflow to the outflow. At the interrupter, the flow experienced the most irregular flow patterns in the pump. The flow at the inflow region in both the impeller and side channel passage rendered weak pressure fluctuation intensities. All three pump cases operated with 24 blades but after one complete circulatory cycle, cases 1, 2 and 3 revealed 21, 20 and 19 regular pressure fluctuations respectively in the impeller flow passage. On the other hand, the side channel flow passage rather produced 24 regular pressure fluctuations. Furthermore, the main frequency harmonic excitations for all studied monitoring points in the impeller and side channel flow passages of the three pump cases occurred at 600 Hz (24 × fn), 1200 Hz (48 × fn), and 1800 Hz (72 × fn). For this reason, exchanged flow times between the impeller and side channel is mainly responsible for the pressure fluctuation which subsequently affects the noise and vibration generation in the side channel pump. Hence, the results could be used as a reference for Noise-Vibration-Harshness (NVH) study in turbomachinery especially modifying the side channel pump in order to improve the operational reliabilities for many engineering processes.
On-Board Energy Storage Devices with Supercapacitors for Metro Trains—Case Study Analysis of Application Effectiveness
Authors: Radu Petru Valentin , Szelag Adam , Steczek Marcin
Published: April 2019

This paper presents an analysis on using an on-board energy storage device (ESD) for enhancing braking energy re-use in electrified railway transportation. A simulation model was developed in the programming language C++ to help with the sizing of the ESD. The simulation model based on the mathematical description has been proposed for a train equipped with on-board ESD for analysis of effectiveness of its application. A case study was carried out for a metro line taking into consideration train characteristics, track alignment, line velocity limits and a running time table. This case study was used to assess the energy savings and perform a cost-benefit analysis for different sizes of the on-board ESD by applying the proposed approach. It was shown that when additional environmental benefits (reduction of CO2 emissions) are considered, this may significantly improve effectiveness of the investments due to CO2 European Emission allowances.
Microgrid Group Trading Model and Solving Algorithm Based on Blockchain
Authors: Xu Zixiao , Yang Dechang , Li Weilin
Published: April 2019

With the development of the energy Internet and the integration of multi-type energy situations, it is of great significance to study the competition game of a multi-agent microgrid group system for its development. As an emerging distributed database technology, blockchain technology has great application potential in the field of energy trading. Firstly, blockchain technology is coupled with the microgrid group transaction, and the information flow transaction model of a microgrid group based on blockchain technology is established. Aiming at this complex multi-objective optimization problem, an improved ant colony optimization algorithm is proposed to solve the model. Finally, the competitive trading model and solving algorithm are simulated and analyzed. The relevant results show that the near global optimum price strategy of each time based on the proposed model can effectively balance the efficiency of each subject in the market. In addition, the model ensures that there is no high-income and low-cost phenomenon in the trading process, therefore the security and quality of the market are guaranteed.
Experimental Investigation on Crack Development Characteristics in Shallow Coal Seam Mining in China
Authors: Huang Qingxiang , He Yanpeng , Cao Jian
Published: April 2019

The development of cracks in mining is the scientific basis for the safety and environmental exploitation of shallow multiple-seam. According to the “thickness of coal seam, interactive distance, and buried depth,” four mining coal mines are selected in Shen Fu-Dong Sheng coalfield (SFDSC). To research the mining conditions of shallow coal seam under different base-load ratio mining conditions and different working faces by the physics simulation and in-sit measurement, the key roof caves are sketched by different colors. This study shows that the typical shallow coal seams in the thin overlying bedrock and thick loose sand layer (LSL) as well as the development of the setup entry cracks (SEC) is dominated by LSL arch damage. The surface cracks are almost directly above the setup entry. The flat seam mining and the SEC development is dominated by parabolic type. The surface cracks are located inside the setup entry. With the mining height increased typically in a shallow coal seam, the rate of crack development and the extent of damaged area increased significantly. The SEC and boundary cracks are fixed. The dynamic periodic cracks (DPC) show the ability of the strata to self-repair. During the multiple-seam mining, the above three kinds of cracks have the phenomenon of activation and development. Through the reasonable coal pillar distance arrangement, the development of boundary cracks can be effectively controlled and the relatively uniform surface settlement and crack closure can be achieved. The purpose of reducing damage mining can also be achieved. Furthermore, it provides scientific support for the green mining in the shallow coal seam.
Identification and Visualization of the Full-Ring Deformation Characteristics of a Large Stormwater Sewage and Storage Tunnel Using Terrestrial Laser Scanning Technology
Authors: Zhang Zixin , Yin Tong , Huang Xin , Zhang Fan , Zhu Yeting , Liu Wei
Published: April 2019

Constructing deeply-buried stormwater sewage and storage tunnels is an effective method to mitigate the waterlogging and sewer overflow problems in modern cities. Prior to construction of such tunnels, a structural loading test is essential for acquiring the mechanical responses under complex loading conditions, such as cyclic inner hydraulic head, during which capturing the full-ring deformation of the tunnel lining is significant for a comprehensive understanding of the tunnel’s mechanical behaviors. This paper introduces the application of terrestrial laser scanning (TLS) technology in the full-scale structural loading tests of a large stormwater sewage and storage tunnel, which gives the full-ring deformation throughout the tests. A data processing methodology was developed to extract the key data points of the lining segments from the original data cloud by removing noise points and mitigating data jump, based on which the deformation of testing the lining segments at arbitrary locations can be calculated. Furthermore, a post-processing software was developed to visualize the full-ring deformation. The full-ring deformation at different loading conditions and its evolution under cyclic loading were captured. It is shown that the lining’s convergence deformation is more sensitive to the inner hydraulic head than to the external soil-water pressure, and the deformation cannot fully recover in a water-inflow-and-drainage cycle due to the presence of joints.
Perinatal Lead (Pb) Exposure and Cortical Neuron-Specific DNA Methylation in Male Mice
Authors: Dou John F. , Farooqui Zishaan , Faulk Christopher D. , Barks Amanda K. , Jones Tamara , Dolinoy Dana C. , Bakulski Kelly M.
Published: April 2019

Lead (Pb) exposure is associated with a wide range of neurological deficits. Environmental exposures may impact epigenetic changes, such as DNA methylation, and can affect neurodevelopmental outcomes over the life-course. Mating mice were obtained from a genetically invariant C57BL/6J background agouti viable yellow Avy strain. Virgin dams (a/a) were randomly assigned 0 ppm (control), 2.1 ppm (low), or 32 ppm (high) Pb-acetate water two weeks prior to mating with male mice (Avy/a), and this continued through weaning. At age 10 months, cortex neuronal nuclei were separated with NeuN+ antibodies in male mice to investigate neuron-specific genome-wide promoter DNA methylation using the Roche NimbleGen Mouse 3x720K CpG Island Promoter Array in nine pooled samples (three per dose). Several probes reached p-value < 10−5, all of which were hypomethylated: 12 for high Pb (minimum false discovery rate (FDR) = 0.16, largest intensity ratio difference = −2.1) and 7 for low Pb (minimum FDR = 0.56, largest intensity ratio difference = −2.2). Consistent with previous results in bulk tissue, we observed a weak association between early-life exposure to Pb and DNA hypomethylation, with some affected genes related to neurodevelopment or cognitive function. Although these analyses were limited to males, data indicate that non-dividing cells such as neurons can be carriers of long-term epigenetic changes induced in development.
Effect of Nanoparticles Surface Charge on the Arabidopsis thaliana (L.) Roots Development and Their Movement into the Root Cells and Protoplasts
Authors: Milewska-Hendel Anna , Zubko Maciej , Stróż Danuta , Kurczyńska Ewa U.
Published: April 2019

Increasing usage of gold nanoparticles (AuNPs) in different industrial areas inevitably leads to their release into the environment. Thus, living organisms, including plants, may be exposed to a direct contact with nanoparticles (NPs). Despite the growing amount of research on this topic, our knowledge about NPs uptake by plants and their influence on different developmental processes is still insufficient. The first physical barrier for NPs penetration to the plant body is a cell wall which protects cytoplasm from external factors and environmental stresses. The absence of a cell wall may facilitate the internalization of various particles including NPs. Our studies have shown that AuNPs, independently of their surface charge, did not cross the cell wall of Arabidopsis thaliana (L.) roots. However, the research carried out with using light and transmission electron microscope revealed that AuNPs with different surface charge caused diverse changes in the root’s histology and ultrastructure. Therefore, we verified whether this is only the wall which protects cells against particles penetration and for this purpose we used protoplasts culture. It has been shown that plasma membrane (PM) is not a barrier for positively charged (+) AuNPs and negatively charged (−) AuNPs, which passage to the cell.
Methylation of SPARCL1 Is Associated with Oncologic Outcome of Advanced Upper Urinary Tract Urothelial Carcinoma
Authors: Luo Hao-Lun , Chiang Po-Huang , Huang Chun-Chieh , Su Yu-Li , Sung Min-Tse , Tsai Eing-Mei , Lin Chang-Shen , Chiang Po-Hui
Published: April 2019

Advanced upper urinary tract urothelial carcinoma (UTUC) is often associated with poor oncologic outcomes. The secreted protein acidic and rich in cysteine-like 1 (SPARCL1) protein, belongs to the SPARC-related family of matricellular proteins. Much literature has been published describing the role of SPARCL1 in the prognosis many cancers. In this study, methylated promoter regions in high-grade and high-stage upper urinary urothelial tumours compared with normal urothelium were analyzed and revealed that SPARCL1 was the most significantly hypermethylated gene in UTUC tissues. Then we prospectively collected UTUC samples and adjacent normal urothelium for pyrosequencing validation, identifying significant CpG site methylation in UTUC tissues. In addition, SPARCL1 RNA levels were significantly lower in UTUC samples. Multivariate Cox regression analysis from 78 patients with solitary renal pelvic or ureteral pT3N0M0 urothelial carcinomas revealed that only negative SPARCL1 expression and nonpapillary tumour architecture were independently associated with systemic recurrence (p = 0.011 and 0.008, respectively). In vitro studies revealed that the behaviour of BFTC-909 cells was less aggressive and more sensitive to radiation or chemotherapy after SPARCL1 overexpression. Thus, SPARCL1 could be considered as a prognostic marker and help decision-making in clinical practice.
Hyaluronic Acid Combined with Serum Rich in Growth Factors in Corneal Epithelial Defects
Authors: Suárez-Barrio Carlota , Etxebarria Jaime , Hernáez-Moya Raquel , del Val-Alonso Marina , Rodriguez-Astigarraga Maddalen , Urkaregi Arantza , Freire Vanesa , Morales María-Celia , Durán Juan Antonio , Vicario Marta , Molina Irene , Herrero-Vanrell Rocío , Andollo Noelia
Published: April 2019

The aim of this study is to assess if an adhesive biopolymer, sodium hyaluronate (NaHA), has synergistic effects with s-PRGF (a serum derived from plasma rich in growth factors and a blood derivative that has already shown efficacy in corneal epithelial wound healing), to reduce time of healing or posology. In vitro proliferation and migration studies, both in human corneal epithelial (HCE) cells and in rabbit primary corneal epithelial (RPCE) cultures, were carried out. In addition, we performed studies of corneal wound healing in vivo in rabbits treated with s-PRGF, NaHA, or the combination of both. We performed immunohistochemistry techniques (CK3, CK15, Ki67, ß4 integrin, ZO-1, α-SMA) in rabbit corneas 7 and 30 days after a surgically induced epithelial defect. In vitro results show that the combination of NaHA and s-PRGF offers the worst proliferation rates in both HCE and RPCE cells. Addition of NaHA to s-PRGF diminishes the re-epithelializing capability of s-PRGF. In vivo, all treatments, given twice a day, showed equivalent efficacy in corneal epithelial healing. We conclude that the combined use of s-PRGF and HaNA as an adhesive biopolymer does not improve the efficacy of s-PRGF alone in the wound healing of corneal epithelial defects.
Conditioned Medium from Human Mesenchymal Stromal Cells: Towards the Clinical Translation
Authors: Sagaradze Georgy , Grigorieva Olga , Nimiritsky Peter , Basalova Nataliya , Kalinina Natalia , Akopyan Zhanna , Efimenko Anastasia
Published: April 2019

Mesenchymal stem/stromal cells (MSC) remain a promising tool for regenerative medicine as the efficacy of MSC-based cell therapy has been demonstrated for a broad spectrum of indications. Their therapeutic potency is mainly associated with their ability to secrete multiple factors critical for tissue regeneration. Due to comparable effects along with superior safety MSC conditioned medium (MSC-CM) containing a complex of MSC-secreted products is considered a reasonable alternative to cell therapy. However, the lack of standards regulating bioprocessing, use of proper auxiliary materials, and quality control complicates the development of MSC secretome-based therapeutics. In this study, we suggested several approaches addressing these issues. We manufactured 36 MSC-CM samples based on different xeno-free serum-free chemically defined media (DMEM-LG or MSC NutriStem® XF) using original protocols and considered total concentrations of regeneration-associated paracrine factors secreted by human adipose-derived MSC at each time-point of conditioning. Using regression analysis, we retrospectively predicted associations between concentrations of several components of MSC-CM and its biological activity to stimulate human dermal fibroblast and endothelial cell migration in vitro as routine examples of potency assays for cell-based products. We also demonstrated that the cell culture medium might affect MSC-CM biological activity to varying degrees depending on the potency assay type. Furthermore, we showed that regression analysis might help to overcome donor variability. The suggested approaches might be successfully applied for other cell types if their secretome was shown to be promising for application in regenerative medicine.
Influence of Fractionation Methods on Physical and Biological Properties of Injectable Platelet-Rich Fibrin: An Exploratory Study
Authors: Thanasrisuebwong Prakan , Surarit Rudee , Bencharit Sompop , Ruangsawasdi Nisarat
Published: April 2019

Injectable platelet-rich fibrin (i-PRF) has been used as an autografting material to enhance bone regeneration through intrinsic growth factors. However, fractionation protocols used to prepare i-PRF can be varied and the effects of different fractionation protocols are not known. In this study, we investigated the influence of different fractions of i-PRF on the physical and biological properties derived from variations in i-PRF fractionation preparation. The i-PRF samples, obtained from the blood samples of 10 donors, were used to harvest i-PRF and were fractioned into two types. The yellow i-PRF fractionation was harvested from the upper yellow zone, while the red i-PRF fractionation was collected from both the yellow and red zone of the buffy coat. The viscoelastic property measurements, including the clot formation time, α-angle, and maximum clot firmness, were performed by rotational thromboelastometry. The fibrin network was examined using a scanning electron microscope. Furthermore, the concentration of growth factors released, including VEGF, TGF-β1, and PDGF, were quantified using ELISA. A paired t-test with a 95% confidence interval was used. All three viscoelastic properties were statistically significantly higher in the yellow i-PRF compared to the red i-PRF. The scanning electron microscope reviewed more cellular components in the red i-PRF compared to the yellow i-PRF. In addition, the fibrin network of the yellow i-PRF showed a higher density than that in the red i-PRF. There was no statistically significant difference between the concentration of VEGF and TGF-β1. However, at Day 7 and Day 14 PDGF concentrations were statistically significantly higher in the red i-PRF compared to the yellow group. In conclusion, these results showed that the red i-PRF provided better biological properties through the release of growth factors. On the other hand, the yellow i-PRF had greater viscoelastic physical properties. Further investigations into the appropriate i-PRF fractionation for certain surgical procedures are therefore necessary to clarify the suitability for each fraction for different types of regenerative therapy.
A Molecular Dynamics Approach to Explore the Intramolecular Signal Transduction of PPAR-α
Authors: Basith Shaherin , Manavalan Balachandran , Shin Tae Hwan , Lee Gwang
Published: April 2019

Dynamics and functions of the peroxisome proliferator-activated receptor (PPAR)-α are modulated by the types of ligands that bind to the orthosteric sites. While several X-ray crystal structures of PPAR-α have been determined in their agonist-bound forms, detailed structural information in their apo and antagonist-bound states are still lacking. To address these limitations, we apply unbiased molecular dynamics simulations to three different PPAR-α systems to determine their modulatory mechanisms. Herein, we performed hydrogen bond and essential dynamics analyses to identify the important residues involved in polar interactions and conformational structural variations, respectively. Furthermore, betweenness centrality network analysis was carried out to identify key residues for intramolecular signaling. The differences observed in the intramolecular signal flow between apo, agonist- and antagonist-bound forms of PPAR-α will be useful for calculating maps of information flow and identifying key residues crucial for signal transductions. The predictions derived from our analysis will be of great help to medicinal chemists in the design of effective PPAR-α modulators and additionally in understanding their regulation and signal transductions.
A New Subclade of Leptosphaeria biglobosa Identified from Brassica rapa
Authors: Zou Zhongwei , Zhang Xuehua , Parks Paula , du Toit Lindsey J. , Van de Wouw Angela P. , Fernando W. G. Dilantha
Published: April 2019

Blackleg (Phoma stem canker) of crucifers is a globally important disease caused by the ascomycete species complex comprising of Leptosphaeria maculans and Leptosphaeria biglobosa. Six blackleg isolates recovered from Brassica rapa cv. Mizspoona in the Willamette Valley of Oregon were characterized as L. biglobosa based on standard pathogenicity tests and molecular phylogenetic analysis. These isolates were compared to 88 characterized L. biglobosa isolates from western Canada, 22 isolates from Australia, and 6 L. maculans isolates from Idaho, USA using maximum parsimony and distance analysis of phylogenetic trees generated from the ITS rDNA (internal transcribed spacer rDNA) sequence, and the actin and β-tubulin gene sequences. The L. biglobosa isolates derived from B. rapa collected in Oregon formed a separate subclade based on concatenated gene sequences or a single gene sequence, regardless of the analyses. Pathogenicity tests showed that these isolates failed to infect either resistant or susceptible B. napus cultivars, but caused severe symptoms on three B. rapa cultivars (Accession number: UM1113, UM1112, and UM1161), a B. oleracea var. capitata (cabbage) cultivar (Copenhagen Market), and two B. juncea cultivars (CBM, a common brown Mustard, and Forge). These findings demonstrated that the L. biglobosa isolates derived from a B. rapa crop in Oregon were genetically distinct from existing species of L. biglobosa, and constitute a new subclade, herein proposed as L. biglobosa ‘americensis’.
Infiltration of T Cells into a Three-Dimensional Psoriatic Skin Model Mimics Pathological Key Features
Authors: Lorthois Isabelle , Simard Mélissa , Morin Sophie , Pouliot Roxane
Published: April 2019

Psoriasis is an autoimmune chronic dermatosis that is T cell-mediated, characterized by epidermal thickening, aberrant epidermal differentiation and inflammatory infiltrates, with a dominant Th1 and Th17 profile. Additional in vitro models are required to study the complex interactions between activated T cells and skin cells, and to develop new, more effective treatments. We have therefore sought to model this psoriatic inflammation by the generation of tissue-engineered immunocompetent tissues, and we have investigated the response of activated T-cell infiltration in models produced with lesional psoriatic skin cells on major hallmarks of psoriasis. The immunocompetent lesional skin model displayed a delayed onset of epidermal differentiation, an hyperproliferation of the basal keratinocytes, a drastic increase in the secretion of proinflammatory cytokines, and a disturbed expression of key transcription factors, as observed in lesional plaques, suggesting a crucial importance of combining the pathological phenotype of cutaneous cells to T cells in order to generate a relevant model for psoriasis. Finally, we found this skin model to be responsive to methotrexate treatment, making it a valuable tool for drug development.
Genetic Restoration of Heme Oxygenase-1 Expression Protects from Type 1 Diabetes in NOD Mice
Authors: Pogu Julien , Tzima Sotiria , Kollias Georges , Anegon Ignacio , Blancou Philippe , Simon Thomas
Published: April 2019

Antigen-presenting cells (APCs) including dendritic cells (DCs) play a critical role in the development of autoimmune diseases by presenting self-antigen to T-cells. Different signals modulate the ability of APCs to activate or tolerize autoreactive T-cells. Since the expression of heme oxygenase-1 (HO-1) by APCs has been associated with the tolerization of autoreactive T-cells, we hypothesized that HO-1 expression might be altered in APCs from autoimmune-prone non-obese diabetic (NOD) mice. We found that, compared to control mice, NOD mice exhibited a lower percentage of HO-1-expressing cells among the splenic DCs, suggesting an impairment of their tolerogenic functions. To investigate whether restored expression of HO-1 in APCs could alter the development of diabetes in NOD mice, we generated a transgenic mouse strain in which HO-1 expression can be specifically induced in DCs using a tetracycline-controlled transcriptional activation system. Mice in which HO-1 expression was induced in DCs exhibited a lower Type 1 Diabetes (T1D) incidence and a reduced insulitis compared to non-induced mice. Upregulation of HO-1 in DCs also prevented further increase of glycemia in recently diabetic NOD mice. Altogether, our data demonstrated the potential of induction of HO-1 expression in DCs as a preventative treatment, and potential as a curative approach for T1D.
Genome-Wide Analysis and Hormone Regulation of Chitin Deacetylases in Silkworm
Authors: Zhang Ziyu , Yan Jiamin , Liu Qing , Zhang Yuhao , Gong Jing , Hou Yong
Published: April 2019

Chitin deacetylases (CDAs) are a group of enzymes involved in chitin metabolism in insects; they play a critical role in molting, pupation, and the modification of chitin. In this study, we identified several CDAs in the silkworm, Bombyx mori (BmCDA), and investigated the effect of various hormones on their expression in B. mori larvae and embryo cell lines (BmE). Eight genes encoding BmCDAs were identified in the silkworm genome. They showed different expression patterns in different tissues, and were classified into three types based on where they were expressed: the exoskeleton, digestive organs, and genital organs. Moreover, we found that some BmCDAs showed upregulated expression during the molting period, especially during the fourth molting period in larvae. We also verified that the expression of BmCDA1–6 was upregulated by treatment with 20-hydroxyecdysone not only in larvae, but also in BmE cells. Interestingly, juvenile hormone analog treatment also upregulated the expression of some BmCDAs. The overexpression of several transcription factors revealed that the POU transcription factor POUM2 may play a major role in the regulation of BmCDA expression. Finally, the silencing of BmCDA1 and BmCDA2 did not lead to abnormal phenotypes or death, but may have led to delays in silkworm pupation. These results provide important information about lepidopteran insects in terms of chitin deacetylases and the regulation of their expression.
Genetic Modification of Mucor circinelloides to Construct Stearidonic Acid Producing Cell Factory
Authors: Khan Md. Ahsanul Kabir , Yang Junhuan , Hussain Syed Ammar , Zhang Huaiyuan , Garre Victoriano , Song Yuanda
Published: April 2019

Stearidonic acid (SDA; 18:4, n-3) is the delta 15-desaturase product of gamma linolenic acid (GLA; 18:3, n-6) and delta 6-desaturase product of alpha linolenic acid (ALA; 18:3, n-3). Construction of engineered oleaginous microbes have been attracting significant interest in producing SDA because of its nutritional value and pharmaceutical applications. Mucor circinelloides is a GLA producing filamentous fungus, which can be a useful tool to produce SDA. This study has, therefore, overexpressed the delta-15 desaturase (D15D) gene from Mortierella alpina in this fungus to construct a SDA-producing cell factory. To produce SDA in M. circinelloides, the homologous overexpression of D15D gene was analyzed. When the gene was overexpressed in M. circinelloides CBS 277.49, up to 5.0% SDA was accumulated in this strain. According to current knowledge, this is the first study describing the construction of a SDA-producing cell factory by overexpression of D15D gene in oleaginous fungus M. circinelloides. A new scope for further research has been established by this work to improve SDA production in this fungus, specifically in its high lipid-producing strain, WJ11.
Rab39a and Rab39b Display Different Intracellular Distribution and Function in Sphingolipids and Phospholipids Transport
Authors: Gambarte Tudela Julián , Buonfigli Julio , Luján Agustín , Alonso Bivou Mariano , Cebrián Ignacio , Capmany Anahí , Damiani María
Published: April 2019

Rab GTPases define the identity and destiny of vesicles. Some of these small GTPases present isoforms that are expressed differentially along developmental stages or in a tissue-specific manner, hence comparative analysis is difficult to achieve. Here, we describe the intracellular distribution and function in lipid transport of the poorly characterized Rab39 isoforms using typical cell biology experimental tools and new ones developed in our laboratory. We show that, despite their amino acid sequence similarity, Rab39a and Rab39b display non-overlapping intracellular distribution. Rab39a localizes in the late endocytic pathway, mainly at multivesicular bodies. In contrast, Rab39b distributes in the secretory network, at the endoplasmic reticulum/cis-Golgi interface. Therefore, Rab39a controls trafficking of lipids (sphingomyelin and phospholipids) segregated at multivesicular bodies, whereas Rab39b transports sphingolipids biosynthesized at the endoplasmic reticulum-Golgi factory. Interestingly, lyso bis-phosphatidic acid is exclusively transported by Rab39a, indicating that both isoforms do not exert identical functions in lipid transport. Conveniently, the requirement of eukaryotic lipids by the intracellular pathogen Chlamydia trachomatis rendered useful for dissecting and distinguishing Rab39a- and Rab39b-controlled trafficking pathways. Our findings provide comparative insights about the different subcellular distribution and function in lipid transport of the two Rab39 isoforms.
A Transcriptomic Insight into the Impact of Colon Cancer Cells on Mast Cells
Authors: Yu Yingxin , Blokhuis Bart R. , Garssen Johan , Redegeld Frank A.
Published: April 2019

Mast cells (MCs) are one of the first immune cells recruited to a tumor. It is well recognized that MCs accumulate in colon cancer lesion and their density is associated with the clinical outcomes. However, the molecular mechanism of how colon cancer cells may modify MC function is still unclear. In this study, primary human MCs were generated from CD34+ progenitor cells and a 3D coculture model was developed to study the interplay between colon cancer cells and MCs. By comparing the transcriptomic profile of colon cancer-cocultured MCs versus control MCs, we identified a number of deregulated genes, such as MMP-2, VEGF-A, PDGF-A, COX2, NOTCH1 and ISG15, which contribute to the enrichment of cancer-related pathways. Intriguingly, pre-stimulation with a TLR2 agonist prior to colon cancer coculture induced upregulation of multiple interferon-inducible genes as well as MHC molecules in MCs. Our study provides an alternative approach to study the influence of colon cancer on MCs. The transcriptome signature of colon cancer-cocultured MCs may potentially reflect the mechanism of how colon cancer cells educate MCs to become pro-tumorigenic in the initial phase and how a subsequent inflammatory signal—e.g., TLR2 ligands—may modify their responses in the cancer milieu.
Therapeutic Approaches with Intravitreal Injections in Geographic Atrophy Secondary to Age-Related Macular Degeneration: Current Drugs and Potential Molecules
Authors: Nebbioso Marcella , Lambiase Alessandro , Cerini Alberto , Limoli Paolo Giuseppe , La Cava Maurizio , Greco Antonio
Published: April 2019

The present review focuses on recent clinical trials that analyze the efficacy of intravitreal therapeutic agents for the treatment of dry age-related macular degeneration (AMD), such as neuroprotective drugs, and complement inhibitors, also called immunomodulatory or anti-inflammatory agents. A systematic literature search was performed to identify randomized controlled trials published prior to January 2019. Patients affected by dry AMD treated with intravitreal therapeutic agents were included. Changes in the correct visual acuity and reduction in geographic atrophy progression were evaluated. Several new drugs have shown promising results, including those targeting the complement cascade and neuroprotective agents. The potential action of the two groups of drugs is to block complement cascade upregulation of immunomodulating agents, and to prevent the degeneration and apoptosis of ganglion cells for the neuroprotectors, respectively. Our analysis indicates that finding treatments for dry AMD will require continued collaboration among researchers to identify additional molecular targets and to fully interrogate the utility of pluripotent stem cells for personalized therapy.
Regulation of Proliferation, Differentiation and Functions of Osteoblasts by Runx2
Authors: Komori Toshihisa
Published: April 2019

Runx2 is essential for osteoblast differentiation and chondrocyte maturation. During osteoblast differentiation, Runx2 is weakly expressed in uncommitted mesenchymal cells, and its expression is upregulated in preosteoblasts, reaches the maximal level in immature osteoblasts, and is down-regulated in mature osteoblasts. Runx2 enhances the proliferation of osteoblast progenitors by directly regulating Fgfr2 and Fgfr3. Runx2 enhances the proliferation of suture mesenchymal cells and induces their commitment into osteoblast lineage cells through the direct regulation of hedgehog (Ihh, Gli1, and Ptch1), Fgf (Fgfr2 and Fgfr3), Wnt (Tcf7, Wnt10b, and Wnt1), and Pthlh (Pthr1) signaling pathway genes, and Dlx5. Runx2 heterozygous mutation causes open fontanelle and sutures because more than half of the Runx2 gene dosage is required for the induction of these genes in suture mesenchymal cells. Runx2 regulates the proliferation of osteoblast progenitors and their differentiation into osteoblasts via reciprocal regulation with hedgehog, Fgf, Wnt, and Pthlh signaling molecules, and transcription factors, including Dlx5 and Sp7. Runx2 induces the expression of major bone matrix protein genes, including Col1a1, Spp1, Ibsp, Bglap2, and Fn1, in vitro. However, the functions of Runx2 in differentiated osteoblasts in the expression of these genes in vivo require further investigation.