Search results for: head and neck squamous cell carcinoma

Authors: Ilzira A. Minigalieva, Tatiana V. Bushueva, Eleonore Frohlich, Vladimir Panov, Ekaterina Shishkina, Boris A. Katsnelson

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Background: The overwhelming majority of the experimental studies in the field of metal nanotoxicology have been performed on cultures of established cell lines, with very few researchers focusing on animal experiments, while a juxtaposition of conclusions inferred from these two types of research is blatantly lacking. The least studied aspect of this problem relates to characterizing and predicting the combined toxicity of metallic nanoparticles. Methods: Comparative and combined toxic effects of purposefully prepared spherical NiO and Mn₃O₄ nanoparticles (mean diameters 16.7 ± 8.2 nm and 18.4 ± 5.4 nm respectively) were estimated on cultures of human cell lines: MRC-5 fibroblasts, THP-1 monocytes, SY-SY5Y neuroblastoma cells, as well as on the latter two lines differentiated to macrophages and neurons, respectively. The combined cytotoxicity was mathematically modeled using the response surface methodology. Results: The comparative assessment of the studied NPs unspecific toxicity previously obtained in vivo was satisfactorily reproduced by the present in vitro tests. However, with respect to manganese-specific brain damage which had been demonstrated by us in animal experiment with the same NPs, the testing on neuronall cell culture showed only a certain enhancing effect of Mn₃O₄-NPs on the toxic action of NiO-NPs, while the role of the latter prevailed. Conclusion: From the point of view of the preventive toxicology, the experimental modeling of metallic NPs combined toxicity on cell cultures can give non-reliable predictions of the in vivo action’s effects.

Keywords: manganese oxide, nickel oxide, nanoparticles, in vitro toxicity

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Authors: Patrizia Frontera, Anastasia Macario, Simona Crispi, Angela Malara, Pierantonio De Luca, Stefano Trocino

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The exploration of solar energy for the photoelectrochemical splitting of water into hydrogen and oxygen has been extensively researched as a means of generating sustainable H₂ fuel. However, despite these efforts, commercialization of this technology has not yet materialized. Presently, the primary impediments to commercialization include low solar-to-hydrogen efficiency (2-3% in PEC with an active area of up to 10-15 cm²), the utilization of costly and critical raw materials (e.g., BiVO₄), and energy losses during the separation of H₂ from O₂ and H₂O vapours in the output stream. The SERGIO partners have identified an advanced approach to fabricate photoelectrode materials, coupled with an appropriate scientific direction to achieve cost-effective solar-driven H₂ production in a tandem photoelectrochemical cell. This project is designed to reach Technology Readiness Level (TRL) 4 by validating the technology in the laboratory using a cell with an active area of up to 10 cm², boasting a solar-to-hydrogen efficiency of 5%, and ensuring acceptable hydrogen purity (99.99%). Our objectives include breakthroughs in cost efficiency, conversion efficiency, and H₂ purity.

Keywords: photoelectrolysis, green hydrogen, photoelectrochemical cell, semiconductors

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Authors: Yuanhao Gao, Ping Lin, Kees Weijer

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An optimal control system model is proposed for the cell flow in the process of chick embryo gastrulation in this paper. The target is to determine the cellular interaction forces which are hard to measure. This paper will take an approach to investigate the forces with the idea of the inverse problem. By choosing the forces as the control variable and regarding the cell flow as Stokes fluid, an objective functional will be established to match the numerical result of cell velocity with the experimental data. So that the forces could be determined by minimizing the objective functional. The Lagrange multiplier method is utilized to derive the state and adjoint equations consisting the optimal control system, which specifies the first-order necessary conditions. Finite element method is used to discretize and approximate equations. A conjugate gradient algorithm is given for solving the minimum solution of the system and determine the forces.

Keywords: optimal control model, Stokes equation, conjugate gradient method, finite element method, chick embryo gastrulation

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Authors: Mohd Tariq

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The three phase system drives produce the problem of more torque pulsations and harmonics. This issue prevents the smooth operation of the drives and it also induces the amount of heat generated thus resulting in an increase in power loss. Higher phase system offers smooth operation of the machines with greater power capacity. Five phase variable-speed induction motor drives are commonly used in various industrial and commercial applications like tractions, electrical vehicles, ship propulsions and conveyor belt drive system. In this work, a comparative analysis of the different modulation schemes applied on the five-level five-phase Packed U Cell (PUC) inverter fed induction motor drives is presented. The performance of the inverter is greatly affected with the modulation schemes applied. The system is modeled, designed, and implemented in MATLAB^®/Simulink environment. Experimental validation is done for the prototype of single phase, whereas five phase experimental validation is proposed in the future works.

Keywords: Packed U-Cell (PUC) inverter, five-phase system, pulse width modulation (PWM), induction motor (IM)

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Authors: Nelson Barrientos, Matthew J. Davies, Marco C. Marques, Darren N. Nesbeth, Gary J. Lye, Nicolas Szita

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Microbioreactor technology is making important advances towards its application in cell culture and bioprocess development. In particular, the technology promises flexible and controllable devices capable to perform parallelised experimentation at low cost. Currently, state of the art methods (e.g. optical sensors) allow the accurate monitoring of the microbioreactor operation. In addition, the laminar flow regime encountered in these devices allows more predictive fluid dynamics modelling, improving the control over the soluble, physical and mechanical environment of the cells. This work describes the development and characterisation of a novel microbioreactor cassette system (microbioreactor volume is 150 μL. The volumetric oxygen transfer coefficient (KLa) and mixing time have been characterised to be between 25 to 113 h-1 and 0.5 and 0.1 s, respectively. In addition, the Residence time distribution (RTD) analysis confirms that the reactor operates at well mixed conditions. Finally, Staphylococcus carnosus TM300 growth is demonstrated via batch culture experiments. Future work consists in expanding the optics of the microbioreactor design to include the monitoring of variables such as fluorescent protein expression, among others.

Keywords: microbioreactor, cell-culture, fermentation, microfluidics

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Authors: Bing Li, Zhi Li, Yilong Yang

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Web classification can promote the quality of service discovery and management in the service repository. It is widely used to locate developers desired services. Although traditional classification methods based on supervised learning models can achieve classification tasks, developers need to manually mark web services, and the quality of these tags may not be enough to establish an accurate classifier for service classification. With the doubling of the number of web services, the manual tagging method has become unrealistic. In recent years, the attention mechanism has made remarkable progress in the field of deep learning, and its huge potential has been fully demonstrated in various fields. This paper designs a multi-head attention graph convolutional network (MHAGCN) service classification method, which can assign different weights to the neighborhood nodes without complicated matrix operations or relying on understanding the entire graph structure. The framework combines the advantages of the attention mechanism and graph convolutional neural network. It can classify web services through automatic feature extraction. The comprehensive experimental results on a real dataset not only show the superior performance of the proposed model over the existing models but also demonstrate its potentially good interpretability for graph analysis.

Keywords: attention mechanism, graph convolutional network, interpretability, service classification, service discovery

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Authors: Nunthawan Nowwarote, Waleerat Sukarawan, Prasit Pavasant, Thanaphum Osathanon

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Interleukin 6 (IL-6) is a multifunctional cytokine, regulating various biological responses in several tissues. A Recent study shows that IL-6 plays a role in stemness maintenance in stem cells isolated from human exfoliated deciduous teeth (SHEDs). However, the role of IL-6 on cell differentiation in SHEDs remains unknown. The present study investigated the effect of IL-6 on SHEDs differentiation. Cells were isolated from dental pulp tissues of human deciduous teeth. Flow cytometry was used to determined mesenchymal stem cell marker expression, and the multipotential differentiation (osteogenic, adipogenic and neurogenic lineage ) was also determined. The mRNA was determined using real-time quantitative polymerase chain reaction, and the phenotypes were confirmed by chemical and immunofluorescence staining. Results demonstrated that SHEDs expressed CD44, CD73, CD90, CD105 but not CD45. Further, the up-regulation of osteogenic, adipogenic and neurogenic marker genes was observed upon maintaining cells in osteogenic, adipogenic and neurogenic induction medium, respectively. The addition of IL-6 induced osteogenic by up-regulated osteogenic marker gene also increased in vitro mineralization. Under neurogenic medium supplement with IL-6, up-regulated neurogenic marker. Whereas, an addition of IL-6 attenuated adipogenic differentiation by SHEDs. In conclusion, this evidence implies that IL-6 may participate in cells differentiation ability of SHEDs.

Keywords: SHEDs, IL-6, cell differentiations, dental pulp

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Authors: Lethukuthula Ngobese, Abin Gupthar, Patrick Govender

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The ability of yeast cell wall-derived mannoproteins (glycoproteins) to positively contribute to oenological properties has been a key factor that stimulates research initiatives into these industrially important glycoproteins. In addition, and from a fundamental research perspective, yeast cell wall glycoproteins are involved in a wide range of biological interactions. To date, and to the best of our knowledge, our understanding of the fine molecular structure of these mannoproteins is fairly limited. Generally, the amino acid sequences of their protein moieties have been established from structural and functional analysis of the genomic sequence of these yeasts whilst far less information is available on the glycosyl moieties of these mannoproteins. A novel strategy was devised in this study that entails the genetic engineering of yeast strains that over-express and release cell wall-associated glycoproteins into the liquid growth medium. To this end, the Flo1p mannoprotein was overexpressed in Saccharomyces cerevisiae laboratory strains bearing a specific deletion in KNR4 and GPI7 genes involved in cell wall biosynthesis that have been previously shown to extracellularly hyper-secrete cell wall-associated glycoproteins. A polymerase chain reaction (PCR) -based cloning strategy was employed to generate transgenic yeast strains in which the native cell wall FLO1 glycoprotein-encoding gene is brought under transcriptional control of the constitutive PGK1 promoter. The modified Helm’s flocculation assay was employed to assess flocculation intensities of a Flo1p over-expressing wild type and deletion mutant as an indirect measure of their abilities to release the desired mannoprotein. The flocculation intensities of the transformed strains were assessed and all the strains showed similar intensities (>98% flocculation). To assess if mannoproteins were released into the growth medium, the supernatant of each strain was subjected to the BCA protein assay and the transformed Δknr4 strain showed a considerable increase in protein levels. This study has the potential to produce mannoproteins in sufficient quantities that may be employed in future investigations to understand their molecular structures and mechanisms of interaction to the benefit of both fundamental and industrial applications.

Keywords: glycoproteins, genetic engineering, flocculation, over-expression

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Authors: William Kelly, Sorelle Veigne, Xianhua Li, Zuyi Huang, Shyamsundar Subramanian, Eugene Schaefer

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The ambr15™ bioreactor is a single-use microbioreactor for cell line development and process optimization. The ambr system offers fully automatic liquid handling with the possibility of fed-batch operation and automatic control of pH and oxygen delivery. With operating conditions for large scale biopharmaceutical production properly scaled down, micro bioreactors such as the ambr15™ can potentially be used to predict the effect of process changes such as modified media or different cell lines. In this study, gassing rates and dilution rates were varied for a semi-continuous cell culture system in the ambr15™ bioreactor. The corresponding changes to metabolite production and consumption, as well as cell growth rate and therapeutic protein production were measured. Conditions were identified in the ambr15™ bioreactor that produced metabolic shifts and specific metabolic and protein production rates also seen in the corresponding larger (5 liter) scale perfusion process. A Dynamic Flux Balance model was employed to understand and predict the metabolic changes observed. The DFB model-predicted trends observed experimentally, including lower specific glucose consumption when CO₂ was maintained at higher levels (i.e. 100 mm Hg) in the broth. A Computational Fluid Dynamic (CFD) model of the ambr15™ was also developed, to understand transfer of O₂ and CO₂ to the liquid. This CFD model predicted gas-liquid flow in the bioreactor using the ANSYS software. The two-phase flow equations were solved via an Eulerian method, with population balance equations tracking the size of the gas bubbles resulting from breakage and coalescence. Reasonable results were obtained in that the Carbon Dioxide mass transfer coefficient (kLa) and the air hold up increased with higher gas flow rate. Volume-averaged kLa values at 500 RPM increased as the gas flow rate was doubled and matched experimentally determined values. These results form a solid basis for optimizing the ambr15™, using both CFD and FBA modelling approaches together, for use in microscale simulations of larger scale cell culture processes.

Keywords: cell culture, computational fluid dynamics, dynamic flux balance analysis, microbioreactor

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Authors: Khaoula Bensaida, Osama Eljamal

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The electrical energy generation through Microbial Fuel Cells (MFCs) using microorganisms is a renewable and sustainable approach. It creates truly an efficient technology for power production and wastewater treatment. MFC is an electrochemical device which turns wastewater into electricity. The most important part of MFC is microbes. Nano zero-valent Iron NZVI technique was successfully applied in degrading the chemical pollutants and cleaning wastewater. However, the use of NZVI for enhancing the current production is still not confirmed yet. This study aims to confirm the effect of these particles on the current generation by using MFC. A constructed microbial fuel cell, which utilizes domestic wastewater, has been considered for wastewater treatment and bio-electricity generation. The two electrodes were connected to an external resistor (200 ohms). Experiments were conducted in two steps. First, the MFC was constructed without adding NZVI particles (Control) while at a second step, nanoparticles were added with a concentration of 50mg/L. After 20 hours, the measured voltage increased to 5 and 8mV, respectively. To conclude, the use of zero-valent iron in an MFC system can increase electricity generation.

Keywords: bacterial growth, electricity generation, microbial fuel cell MFC, nano zero-valent iron NZVI.

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Authors: Murali Aarthy, Sanjeev Kumar Singh

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High risk human papillomaviruses are highly associated with the carcinoma of the cervix and the other genital tumors. Cervical cancer develops through the multistep process in which increasingly severe premalignant dysplastic lesions called cervical intraepithelial neoplastic progress to invasive cancer. The oncoprotein E7 of human papillomavirus expressed in the lower epithelial layers drives the cells into S-phase creating an environment conducive for viral genome replication and cell proliferation. The replication of the virus occurs in the terminally differentiating epithelium and requires the activation of cellular DNA replication proteins. To date, no suitable drug molecule is available to treat HPV infection whereas identification of potential drug targets and development of novel anti-HPV chemotherapies with unique mode of actions are expected. Hence, our present study aimed to identify the potential inhibitors analogous to EGCG, a green tea molecule which is considered to be safe to use for mammalian systems. A 3D similarity search on the natural small molecule library from natural product database using EGCG identified 11 potential hits based on their similarity score. The structure based docking strategies were implemented in the potential hits and the key interacting residues of protein with compounds were identified through simulation studies and binding free energy calculations. The conformational changes between the apoprotein and the complex were analyzed with the simulation and the results demonstrated that the dynamical and structural effects observed in the protein were induced by the compounds and indicated the dominance to the oncoprotein. Overall, our study provides the basis for the structural insights of the identified potential hits and EGCG and hence, the analogous compounds identified can be potent inhibitors against the HPV 16 E7 oncoprotein.

Keywords: EGCG, oncoprotein, molecular dynamics simulation, analogues

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Authors: Adhimoolam Saravana Kumar, K. N. Govindarajan, B. Devanand, R. Rajakumar

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The quality of Computed Tomography (CT) procedures has improved in recent years due to technological developments and increased diagnostic ability of CT scanners. Due to the fact that CT doses are the peak among diagnostic radiology practices, it is of great significance to be aware of patient’s CT radiation dose whenever a CT examination is preferred. CT radiation dose delivered to patients in the form of volume CT dose index (CTDIvol) values, is displayed on scanner monitors at the end of each examination and it is an important fact to assure that this information is accurate. The objective of this study was to estimate the CTDIvol values for great number of patients during the most frequent CT examinations, to study the comparison between CT dose monitor values and measured ones, as well as to highlight the fluctuation of CTDIvol values for the same CT examination at different centres and scanner models. The output CT dose indices measurements were carried out on single and multislice scanners for available kV, 5 mm slice thickness, 100 mA and FOV combination used. The 100 CT scanners were involved in this study. Data with regard to 15,000 examinations in patients, who underwent routine head, chest and abdomen CT were collected using a questionnaire sent to a large number of hospitals. Out of the 15,000 examinations, 5000 were head CT examinations, 5000 were chest CT examinations and 5000 were abdominal CT examinations. Comprehensive quality assurance (QA) was performed for all the machines involved in this work. Followed by QA, CT phantom dose measurements were carried out in South India using actual scanning parameters used clinically by the hospitals. From this study, we have measured the mean divergence between the measured and displayed CTDIvol values were 5.2, 8.4, and -5.7 for selected head, chest and abdomen procedures for protocols as mentioned above, respectively. Thus, this investigation revealed an observable change in CT practices, with a much wider range of studies being performed currently in South India. This reflects the improved capacity of CT scanners to scan longer scan lengths and at finer resolutions as permitted by helical and multislice technology. Also, some of the CT scanners have used smaller slice thickness for routine CT procedures to achieve better resolution and image quality. It leads to an increase in the patient radiation dose as well as the measured CTDIv, so it is suggested that such CT scanners should select appropriate slice thickness and scanning parameters in order to reduce the patient dose. If these routine scan parameters for head, chest and abdomen procedures are optimized than the dose indices would be optimal and lead to the lowering of the CT doses. In South Indian region all the CT machines were routinely tested for QA once in a year as per AERB requirements.

Keywords: CT dose index, weighted CTDI, volumetric CTDI, radiation dose

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Authors: Guillaume Soubeyran, Fabrice Micoud, Benoit Morin, Jean-Philippe Poirot-Crouvezier, Magali Reytier

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Proton Exchange Membrane Fuel Cell (PEMFC) technology is considered as a solution for the reduction of CO2 emissions. However, this technology still meets several challenges for high-scale industrialization. In this context, the increase of durability remains a critical aspect for competitiveness of this technology. Fortunately, performance degradations in nominal operating conditions is partially reversible, meaning that if specific conditions are applied, a partial recovery of fuel cell performance can be achieved, while irreversible degradations can only be mitigated. Thus, it is worth studying the optimal conditions to rejuvenate these reversible degradations and assessing the long-term impact of such procedures on the performance of the cell. Reversible degradations consist mainly of anode Pt active sites poisoning by carbon monoxide at the anode, heterogeneities in water management during use, and oxidation/deactivation of Pt active sites at the cathode. The latter is identified as a major source of reversible performance loss caused by the presence oxygen, high temperature and high cathode potential that favor platinum oxidation, especially in high efficiency operating points. Hence, we studied here a recovery procedure aiming at reducing the platinum oxides by decreasing cathode potential during operation. Indeed, the application of short air starvation phase leads to a drop of cathode potential. Cell performances are temporarily increased afterwards. Nevertheless, local temperature and current heterogeneities within the cells are favored and shall be minimized. The consumption of fuel during the recovery phase shall also be considered to evaluate the global efficiency. Consequently, the purpose of this work is to find an optimal compromise between the recovery of reversible degradations by air starvation, the increase of global cell efficiency and the mitigation of irreversible degradations effects. Different operating parameters have first been studied such as cell voltage, temperature and humidity in single cell set-up. Considering the global PEMFC system efficiency, tests showed that reducing duration of recovery phase and reducing cell voltage was the key to ensure an efficient recovery. Recovery phase frequency was a major factor as well. A specific method was established to find the optimal frequency depending on the duration and voltage of the recovery phase. Then, long-term degradations have also been studied by applying FC-DLC cycles based on NEDC cycles on a 4-cell short stack by alternating test sequences with and without recovery phases. Depending on recovery phase timing, cell efficiency during the cycle was increased up to 2% thanks to a mean voltage increase of 10 mV during test sequences with recovery phases. However, cyclic voltammetry tests results suggest that the implementation of recovery phases causes an acceleration of the decrease of platinum active areas that could be due to the high potential variations applied to the cathode electrode during operation.

Keywords: durability, PEMFC, recovery procedure, reversible degradation

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Authors: Chih-Yang Liao

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Baseball is usually played outdoors in the warmest months of the year. Therefore, baseball players are susceptible to the influence of the hot environment. It has been shown that hitting performance is increased in games played in warm weather, compared to in cold weather, in Major League Baseball. Intermittent cooling during sporting events can prevent the risk of hyperthermia and increase endurance performance. However, the effects of cooling during baseball games played in a hot environment are unclear. This study adopted a cross-over design. Ten Division I collegiate male baseball players in Taiwan volunteered to participate in this study. Each player played two simulated baseball games, with one day in between. Five of the players received intermittent cooling during the first simulated game, while the other five players received intermittent cooling during the second simulated game. The participants were covered in neck and forehand regions for 6 min with towels that were soaked in icy salt water 3 to 4 times during the games. The participants received the cooling treatment in the dugout when they were not on the field for defense or hitting. During the 2 simulated games, the temperature was 31.1-34.1°C and humidity was 58.2-61.8%, with no difference between the two games. Ratings of perceived exertion, thermal sensation, tympanic and forehead skin temperature immediately after each defensive half-inning and after cooling treatments were recorded. Ratings of perceived exertion were measured using the Borg 10-point scale. The thermal sensation was measured with a 6-point scale. The tympanic and skin temperature was measured with infrared thermometers. The data were analyzed with a two-way analysis of variance with repeated measurement. The results showed that intermitted cooling significantly reduced ratings of perceived exertion and thermal sensation. Forehead skin temperature was also significantly decreased after cooling treatments. However, the tympanic temperature was not significantly different between the two trials. In conclusion, intermittent cooling in the neck and forehead regions was effective in alleviating the perceived exertion and heat sensation. However, this cooling intervention did not affect the core temperature. Whether intermittent cooling has any impact on hitting or pitching performance in baseball players warrants further investigation.

Keywords: baseball, cooling, ratings of perceived exertion, thermal sensation

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Authors: H. Oudira, A. Saifi

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The therapeutic utility of certain Auger emitters such as iodine-125 depends on their position within the cell nucleus . Or diagnostically, and to maintain as low as possible cell damage, it is preferable to have radionuclide localized outside the cell or at least the core. One solution to this problem is to consider markers capable of conveying anticancer drugs to the tumor site regardless of their location within the human body. The objective of this study is to simulate the impact of a complex such as bleomycin on single and double strand breaks in the DNA molecule. Indeed, this simulation consists of the following transactions: - Construction of BLM -Fe- DNA complex. - Simulation of the electron’s transport from the metastable state excitation of Fe 57 by the Monte Carlo method. - Treatment of chemical reactions in the considered environment by the diffusion equation. For physical, physico-chemical and finally chemical steps, the geometry of the complex is considered as a sphere of 50 nm centered on the binding site , and the mathematical method used is called step by step based on Monte Carlo codes.

Keywords: concentration, yield, radical species, bleomycin, excitation, DNA

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Authors: Pyi Phyo Maung

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In the study, the calculations of the permeability coefficient, values of the volume and porosity of a unit cell of a woven fabric before and after deformation based on the geometrical parameters are presented. Two types of carbon woven fabric structures were investigated: standard type, which integrated the filament, has a cross sectional shape of a cylinder and spread tow type, which has a rectangular cross sectional shape. The space antennas reflector, which distinctive feature is the presence of the surface of double curvature, is considered as the object of the research. Modeling of the kinetics of the process of impregnation of the reflector for the two types of carbon fabric’s unit cell structures was performed using software RAM-RTM. This work also investigated the influence of the grid angle between warp and welt of the unit cell on the duration of impregnation process. The results showed that decreasing the angle between warp and welt of the unit cell, the decreasing of the permeability values were occurred. Based on the results of calculation samples of the reflectors, their quality was determined. The comparisons of the theoretical and experimental results have been carried out. Comparison of the two textile structures (standard and spread tow) showed that the standard textiles with circular cross section were impregnated faster than spread tows, which have a rectangular cross section.

Keywords: vacuum assistant resin infusion, impregnation time, shear angle, reflector and modeling

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Authors: Sara Pereira-Nogueira, Tim Worbs, Marc Permanyer-Bosser, Reinhold Förster

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While the entry mechanism of lymphocytes into the lymph node via the blood are well described, it is still largely unknown how cells enter lymph nodes that arrive via afferent lymphatics. In order to address this, our group has established a micro-injection technique in mice through which cells are delivered directly into the lymphatic vessel immediately afferent to the popliteal lymph node. Injected cells can then be tracked via multi-colour fluorescence or 2-photon microscopy, and their localization can be analysed within the popliteal or downstream lymph nodes by immunohistology. Since naïve B cells express the chemokine receptor CXCR5 we intra-lymphatically co-injected B cells derived from wildtype and Cxcr5-deficient mice. While CXCR5 does not play a role in guiding B cells out of the subcapsular sinus, it affects their positioning within the lymph node parenchyma, since CXCR5-deficient B cells are impaired in migrating into the B cell follicle. The knowledge obtained by studying B-cell migration may prove beneficial in clinical settings regarding tumor metastasis or autoimmune diseases.

Keywords: afferent lymphatics, B cell migration, chemokine, intra-lymphatic injection

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Authors: Fatemeh Abbasi, Arne Hofemeier, Timo Betz

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Muscle growth and regeneration critically depend on satellite cells (SCs) which are muscle stem cells located between the basal lamina and myofibres. Upon damage, SCs become activated, enter the cell cycle, and give rise to myoblasts that form new myofibres, while a sub-population self-renew and re-populate the muscle stem cell niche. In aged muscle as well as in certain muscle diseases such as muscular dystrophy, some of the SCs lose their regenerative ability. Although it is demonstrated that the chemical composition of SCs quiescent niche is different from the activated niche, the mechanism initially activated in the SCs remains unknown. While extensive research efforts focused on potential chemical activation, no such factor has been identified to the author’s best knowledge. However, it is substantiated that niche mechanics affects SCs behaviors, such as stemness and engraftment. We hypothesize that mechanical stress in the healthy niche (homeostasis) is different from the regenerative niche and that this difference could serve as an early signal activating SCs upon fiber damage. To investigate this hypothesis, we develop a biomimetic system to reconstitute both, the mechanical and the chemical environment of the SC niche. Cells will be confined between two elastic polyacrylamide (PAA) hydrogels with controlled elastic moduli and functionalized surface chemistry. By controlling the distance between the PAA hydrogel surfaces, we vary the compression forces exerted by the substrates on the cells, while the lateral displacement of the upper hydrogel will create controlled shear forces. To establish such a system, a simplified system is presented. We engineered a sandwich-like configuration of two elastic PAA layer with stiffnesses between 1 and 10 kPa and confined a precursor myoblast cell line (C2C12) in between these layers. Our initial observations in this sandwich model indicate that C2C12 cells show different behaviors under mechanical compression if compared to a control one-layer gel without compression. Interestingly, this behavior is stiffness-dependent. While the shape of C2C12 cells in the sandwich consisting of two stiff (10 kPa) layers was much more elongated, showing almost a neuronal phenotype, the cell shape in a sandwich situation consisting of one stiff and one soft (1 kPa) layer was more spherical. Surprisingly, even in proliferation medium and at very low cell density, the sandwich situation stimulated cell differentiation with increased striation and myofibre formation. Such behavior is commonly found for confluent cells in differentiation medium. These results suggest that mechanical changes in stiffness and applied pressure might be a relevant stimulation for changes in muscle cell behavior.

Keywords: C2C12 cells, compression, force, satellite cells, skeletal muscle

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Authors: Safa Gharbi, Hayfa Zgaya, Nesrine Zoghlami, Slim Hammadi, Cyril De Barbarin, Laurent Vinatier, Christiane Coupier

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Increasing productivity and quality of service, improving the working comfort and ensuring the efficiency of all processes are important challenges for every warehouse. The order picking is recognized to be the most important and costly activity of all the process in warehouses. This paper presents a new approach using Augmented Reality (AR) in the field of logistics. It aims to create a Head-Up Display (HUD) interface with a Warehouse Management System (WMS), using AR glasses. Integrating AR technology allows the optimization of order picking by reducing time of picking process, increasing the efficiency and delivering quickly. The picker will be able to access immediately to all the information needed for his tasks. All the information is displayed when needed in the field of vision (FOV) of the operator, without any action requested from him. These research works are part of the industrial project RASL (Réalité Augmentée au Service de la Logistique) which gathers two major partners: the LAGIS (Laboratory of Automatics, Computer Engineering and Signal Processing in Lille-France) and Genrix Group, European leader in warehouses logistics, who provided his software for implementation, and his logistics expertise.

Keywords: Augmented Reality (AR), logistics and optimization, Warehouse Management System (WMS), Head-Up Display (HUD)

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Authors: J. E. Cobbinah

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One of the most challenging issues confronting schools is good leadership succession planning. Experts argue that, although the idea of leadership succession planning is one of the strategies or practices that can help sustain improvement and promote continuity of good leadership, seem to have been neglected in many schools over the years. Appointment of head teachers in senior high schools is based on long service or one’s ability to demonstrate his/her competence in a leadership selection interview. There is no clear and well-structured leadership succession plan, before leadership position is filled, while school leadership succession planning seem to be an issue that nobody talks about. In faith-based schools the issue is even worse, because religious groups impose whoever they consider strong in the faith on schools as leaders, irrespective of the individual competence, ability to take up challenges associated with individuals’ preparedness to take up leadership position. Therefore, the present study examined the nature (including type) of leadership succession plans in faith-based senior high schools and its associated challenges. Convergent mixed method design was employed to effectively achieve the objectives of the study. The data collection strategies involved the use of interviews, questionnaires, and reviews of secondary data. The data was gathered from students, school leaders (head teachers, deputy heads, and head of departments), selected parents teachers associated members, school management committee members and members from school governors. The results show that governors of faith-based schools are making efforts to enhance education quality, by making school leadership accountable, the absence and the neglect of clear, and well-structured leadership succession plan has some negative outcomes. Unsustainable students’ academic performance, lack of support from existing staffs and senior leaders and lack of support in the implementation of school improvement plan. It would be concluded that, faith-based schools should focus on leadership competence and abilities in the selection process of potential school leaders to achieve a good succession plan rather than appointing leaders who are affiliates of one’s faith.

Keywords: school leadership, succession planning, faith-based schools, school governors

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Authors: Leo Nnamdi Ozurumba-Dwight

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Single-cell genomic analytical systems have proved to be a platform to isolate bulk cells into selected single cells for genomic, proteomic, and related metabolomic studies. This is enabling systematic investigations of the level of heterogeneity in a diverse and wide pool of cell populations. Single cell technologies, embracing techniques such as high parameter flow cytometry, single-cell sequencing, and high-resolution images are playing vital roles in these investigations on messenger ribonucleic acid (mRNA) molecules and related gene expressions in tracking the nature and course of disease conditions. This entails targeted molecular investigations on unit cells that help us understand cell behavoiur and expressions, which can be examined for their health implications on the health state of patients. One of the vital good sides of single-cell RNA sequencing (scRNA seq) is its probing capacity to detect deranged or abnormal cell populations present within homogenously perceived pooled cells, which would have evaded cursory screening on the pooled cell populations of biological samples obtained as part of diagnostic procedures. Despite conduction of just single-cell transcriptome analysis, scRNAseq now permits comparison of the transcriptome of the individual cells, which can be evaluated for gene expressional patterns that depict areas of heterogeneity with pharmaceutical drug discovery and clinical treatment applications. It is vital to strictly work through the tools of investigations from wet lab to bioinformatics and computational tooled analyses. In the precise steps for scRNAseq, it is critical to do thorough and effective isolation of viable single cells from the tissues of interest using dependable techniques (such as FACS) before proceeding to lysis, as this enhances the appropriate picking of quality mRNA molecules for subsequent sequencing (such as by the use of Polymerase Chain Reaction machine). Interestingly, scRNAseq can be deployed to analyze various types of biological samples such as embryos, nervous systems, tumour cells, stem cells, lymphocytes, and haematopoietic cells. In haematopoietic cells, it can be used to stratify acute myeloid leukemia patterns in patients, sorting them out into cohorts that enable re-modeling of treatment regimens based on stratified presentations. In immunotherapy, it can furnish specialist clinician-immunologist with tools to re-model treatment for each patient, an attribute of precision medicine. Finally, the good predictive attribute of scRNAseq can help reduce the cost of treatment for patients, thus attracting more patients who would have otherwise been discouraged from seeking quality clinical consultation help due to perceived high cost. This is a positive paradigm shift for patients’ attitudes primed towards seeking treatment.

Keywords: immunotherapy, transcriptome, re-modeling, mRNA, scRNA-seq

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Authors: Adhara Puspa Noorita, Azkiyatin Nailil M., Rita Aryanti, Sushanti Nuraini, Pujiati Abbas, Suparmi

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T-Dabuto is a tea made from leaves and fruits of parijoto (Madinella speciosa L.), which flavonoid, saponin and tanin contained in that tea are reported have diarrhea-caused antibacterial activity. However, the in vivo antidiarrhea effect have not clear yet. This study was conducted to determine the effect of T-DABUTO to faecal characteristics in male Balb/C-mice induced oleum ricini. Experimental research with post-test only control group design was conducted using 35 young male mice strain Balb-C which was divided into 5 groups. All groups were induced by 0.7 ml/ head of oleum ricini and 3 hours later followed by aquadest for first group, while the 2nd, 3rd, 4th and 5th group were treated by T-DABUTO solution with 75 mg/kgBW, 150 mg/kgBW, 300 mg/kgBW, and 600 mg/kgBW respectively as 0.7 ml/ head/ 0.5 hous for 8 hours. Feces collected were used to identify the frequency, absorbtion diameter and fecal weight. T-DABUTO on dose 75 mg/kg BW has the highest antidiarrhea activity which the mean of frequency defecation, water feacal absorbsion and feacal weight were 1.71±0.95 times, 0.38±0.49 mm, 0.43±0.28 mg, respectively. The T-DABUTO treatment did not influence the body weight of diarrheal mice. The T-DABUTO is potential as one of natural diarrhea tratment, especially in children.

Keywords: diarrhea, flavonid, tannin, saponin

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Authors: Anthony S. Machi, Joseph Minardi

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We present a case report of left atrial myxoma diagnosed by bedside transesophageal (TEE) ultrasound. Left atrial myxoma is the most common benign cardiac tumor and can obstruct blood flow and cause valvular insufficiency. Common symptoms consist of dyspnea, pulmonary edema and other features of left heart failure in addition to thrombus release in the form of tumor fragments. The availability of bedside ultrasound equipment is essential for the quick diagnosis and treatment of various emergency conditions including cardiac neoplasms. A 48-year-old Caucasian female with a four-year history of an untreated renal mass and anemia presented to the ED with two months of sharp, intermittent, bilateral flank pain radiating into the abdomen. She also reported intermittent vomiting and constipation along with generalized body aches, night sweats, and 100-pound weight loss over last year. She had a CT in 2013 showing a 3 cm left renal mass and a second CT in April 2016 showing a 3.8 cm left renal mass along with a past medical history of diverticulosis, chronic bronchitis, dyspnea on exertion, uncontrolled hypertension, and hyperlipidemia. Her maternal family history is positive for breast cancer, hypertension, and Type II Diabetes. Her paternal family history is positive for stroke. She was a current everyday smoker with an 11 pack/year history. Alcohol and drug use were denied. Physical exam was notable for a Grade II/IV systolic murmur at the right upper sternal border, dyspnea on exertion without angina, and a tender left lower quadrant. Her vitals and labs were notable for a blood pressure of 144/96, heart rate of 96 beats per minute, pulse oximetry of 96%, hemoglobin of 7.6 g/dL, hypokalemia, hypochloremia, and multiple other abnormalities. Physicians ordered a CT to evaluate her flank pain which revealed a 7.2 x 8.9 x 10.5 cm mixed cystic/solid mass in the lower pole of the left kidney and a filling defect in the left atrium. Bedside TEE was ordered to follow up on the filling defect. TEE reported an ejection fraction of 60-65% and visualized a mobile 6 x 3 cm mass in the left atrium attached to the interatrial septum extending into the mitral valve. Cardiothoracic Surgery and Urology were consulted and confirmed a diagnosis of left atrial myxoma and clear cell renal cell carcinoma. The patient returned a week later due to worsening nausea and vomiting and underwent emergent nephrectomy, lymph node dissection, and colostomy due to a necrotic colon. Her condition declined over the next four months due to lung and brain metastases, infections, and other complications until she passed away.

Keywords: bedside ultrasound, echocardiography, emergency medicine, left atrial myxoma

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Authors: Ya L. V., Benjamin Ong Wei Lin, Wanli Niu, Benjamin Seah Chin Tat

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This article introduces a methodology that improves reliability and cyclability of 2nd-life Li-ion battery system repurposed as an energy storage system (ESS). Most of the 2nd-life retired battery systems in the market have module/pack-level state-of-health (SOH) indicator, which is utilized for guiding appropriate depth-of-discharge (DOD) in the application of ESS. Due to the lack of cell-level SOH indication, the different degrading behaviors among various cells cannot be identified upon reaching retired status; in the end, considering end-of-life (EOL) loss and pack-level DOD, the repurposed ESS has to be oversized by > 1.5 times to complement the application requirement of reliability and cyclability. This proposed battery grading algorithm, using non-invasive methodology, is able to detect outlier cells based on historical voltage data and calculate cell-level historical maximum temperature data using semi-analytic methodology. In this way, the individual battery cell in the 2nd-life battery system can be graded in terms of SOH on basis of the historical voltage fluctuation and estimated historical maximum temperature variation. These grades will have corresponding DOD grades in the application of the repurposed ESS to enhance system reliability and cyclability. In all, this introduced battery grading algorithm is non-invasive, compatible with all kinds of retired Li-ion battery systems which lack of cell-level SOH indication, as well as potentially being embedded into battery management software for preventive maintenance and real-time cyclability optimization.

Keywords: battery grading algorithm, 2nd-life repurposing battery system, semi-analytic methodology, reliability and cyclability

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Authors: Ashwini Shripatwar, Mayur Biyani, Nikhil Rao, Rajendra Bodake, Sachin Sane

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This paper documents the correlation of valve cover sealing, cracking, and warpage Finite Element Modelling with observations on engine test development. The valve cover is a component mounted on engine head with a gasket which provides sealing against oil which flows around camshaft, valves, rockers, and other overhead components. Material nonlinearity and contact nonlinearity characteristics are taken into consideration because the valve cover is made of a composite material having temperature dependent elastic-plastic properties and because the gasket load-deformation curve is also nonlinear. The leakage is observed between the valve cover and the engine head due to the insufficient contact pressure. The crack is observed on the valve cover due to force application at a region with insufficient stiffness and with elevated temperature. The valve cover shrinkage is observed during the disassembly process on hot exhaust side bolt holes after the engine has been running. In this paper, an analytical approach is developed to correlate a Finite Element Model with the observed failures and to address the design issues associated with the failure modes in question by making design changes in the model.

Keywords: cracking issue, gasket sealing analysis, nonlinearity of contact and material, valve cover

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Authors: David A. Ullisch, Yantree D. Sankar-Thomas, Stefan Wilke, Thomas Selge, Matthias Pump, Thomas Leibold, Kai Schütte, Gilbert Gorr

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Plants have been considered as a source of natural substances for ages. Secondary metabolites from plants are utilized especially in medical applications but are more and more interesting as cosmetical ingredients and in the field of nutraceuticals. However, supply of compounds from natural harvest can be limited by numerous factors i.e. endangered species, low product content, climate impacts and cost intensive extraction. Especially in the pharmaceutical industry the ability to provide sufficient amounts of product and high quality are additional requirements which in some cases are difficult to fulfill by plant harvest. Whereas in many cases the complexity of secondary metabolites precludes chemical synthesis on a reasonable commercial basis, plant cells contain the biosynthetic pathway – a natural chemical factory – for a given compound. A promising approach for the sustainable production of natural products can be plant cell fermentation (PCF®). A thoroughly accomplished development process comprises the identification of a high producing cell line, optimization of growth and production conditions, the development of a robust and reliable production process and its scale-up. In order to address persistent, long lasting production, development of cryopreservation protocols and generation of working cell banks is another important requirement to be considered. So far the most prominent example using a PCF® process is the production of the anticancer compound paclitaxel. To demonstrate the power of plant suspension cultures here we present three case studies: 1) For more than 17 years Phyton produces paclitaxel at industrial scale i.e. up to 75,000 L in scale. With 60 g/kg dw this fully controlled process which is applied according to GMP results in outstanding high yields. 2) Thapsigargin is another anticancer compound which is currently isolated from seeds of Thapsia garganica. Thapsigargin is a powerful cytotoxin – a SERCA inhibitor – and the precursor for the derivative ADT, the key ingredient of the investigational prodrug Mipsagargin (G-202) which is in several clinical trials. Phyton successfully generated plant cell lines capable to express this compound. Here we present data about the screening for high producing cell lines. 3) The third case study covers ingenol-3-mebutate. This compound is found in the milky sap of the intact plants of the Euphorbiacae family at very low concentrations. Ingenol-3-mebutate is used in Picato® which is approved against actinic keratosis. Generation of cell lines expressing significant amounts of ingenol-3-mebutate is another example underlining the strength of plant cell culture. The authors gratefully acknowledge Inspyr Therapeutics for funding.

Keywords: Ingenol-3-mebutate, plant cell culture, sustainability, thapsigargin

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Authors: Jienny Lee, In-Soo Cho, Sang-Ho Cha

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Adult mesenchymal stem cells (MSCs) have been investigated using preclinical approaches for tissue regeneration. Porcine MSCs (pMSCs) are capable of growing and attaching to plastic with a fibroblast-like morphology and then differentiating into bone, adipose, and cartilage tissues in vitro. This study was conducted to investigate the proliferating abilities, differentiation potentials, and multipotency of miniature pig adipose tissue-derived MSCs (mpAD-MSCs) with or without long-term cryopreservation, considering that cryostorage has the potential for use in clinical applications. After confirming the characteristics of the mpAD-MSCs, we examined the effect of long-term cryopreservation (> 2 years) on expression of cell surface markers (CD34, CD90 and CD105), proliferating abilities (cumulative population doubling level, doubling time, colony-forming unit, and MTT assay) and differentiation potentials into mesodermal cell lineages. As a result, the expression of cell surface markers is similar between thawed and fresh mpAD-MSCs. However, long-term cryopreservation significantly lowered the differentiation potentials (adipogenic, chondrogenic, and osteogenic) of mpAD-MSCs. When compared with fresh mpAD-MSCs, thawed mpAD-MSCs exhibited lower expression of mesodermal cell lineage-related genes such as peroxisome proliferator-activated receptor-g2, lipoprotein lipase, collagen Type II alpha 1, osteonectin, and osteocalcin. Interestingly, long-term cryostoraged mpAD-MSCs exhibited significantly higher cell viability than the fresh mpAD-MSCs. Long-term cryopreservation induced a 30% increase in the cell viability of mpAD-MSCs when compared with the fresh mpAD-MSCs at 5 days after thawing. However, long-term cryopreservation significantly lowered expression of stemness markers such as Oct3/4, Sox2, and Nanog. Furthermore, long-term cryopreservation negatively affected expression of senescence-associated genes such as telomerase reverse transcriptase and heat shock protein 90 of mpAD-MSCs when compared with the fresh mpAD-MSCs. The results from this study might be important for the successful application of MSCs in clinical trials after long-term cryopreservation.

Keywords: mesenchymal stem cells, cryopreservation, stemness, senescence

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Authors: Vineeth Siripuram, Abhineet Nigam

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A bioreactor is an engineered system that supports a biologically active environment. Along the years, the advancements in bioreactors have been widely accepted all over the world for varied applications ranging from sewage treatment to tissue cloning. Driven by tissue and organ shortage, tissue engineering has emerged as an alternative to transplantation for the reconstruction of lost or damaged organs. In this study, Computational fluid dynamics (CFD) has been used to model porous medium flow in scaffolds (taken from the literature) with different flow patterns. A detailed analysis of different scaffold geometries and their influence on cell deposition in the perfusion system is been carried out using Computational fluid dynamics (CFD). Considering the fact that, the scaffold should mimic the organs or tissues structures in a three-dimensional manner, certain assumptions were made accordingly. The research on scaffolds has been extensively carried out in different bioreactors. However, there has been less focus on the morphology of the scaffolds and the flow patterns in which the perfusion system is laid upon. The objective of this paper is to employ a computational approach using CFD simulation to determine the optimal morphology and the anisotropic measurements of the various samples of scaffolds. Using predictive computational modelling approach, variables which exert dominant effects on the cell deposition within the scaffold were prioritised and corresponding changes in morphology of scaffold and flow patterns in the perfusion systems are made. A Eulerian approach was carried on in multiple CFD simulations, and it is observed that the morphological and topological changes in the scaffold perfusion system are of great importance in the commercial applications of scaffolds.

Keywords: cell seeding, CFD, flow patterns, modelling, perfusion systems, scaffold

Procedia PDF Downloads 156

Authors: Vongsathorn Ngampuak, Yutachai Chookaew, Wipawee Dejtisakdi

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Dunaliella salina has great potential as a system for generating commercially valuable products, including beta-carotene, pharmaceuticals, and biofuels. Our goal is to improve this potential by enhancing growth rate and other properties of D. salina under optimal growth conditions. We used ethyl methane sulfonate (EMS) to generate random mutants in D. salina KU11, a strain classified in Thailand. In a preliminary experiment, we first treated D. salina cells with 0%, 0.8%, 1.0%, 1.2%, 1.44% and 1.66% EMS to generate a killing curve. After that, we randomly picked 30 candidates from approximately 300 isolated survivor colonies from the 1.44% EMS treatment (which permitted 30% survival) as an initial test of the mutant screen. Among the 30 survivor lines, we found that 2 strains (mutant #17 and #24) had significantly improved growth rates and cell number accumulation at stationary phase approximately up to 1.8 and 1.45 fold, respectively, 2 strains (mutant #6 and #23) had significantly decreased growth rates and cell number accumulation at stationary phase approximately down to 1.4 and 1.35 fold, respectively, while 26 of 30 lines had similar growth rates compared with the wild type control. We also analyzed cell size for each strain and found there was no significant difference comparing all mutants with the wild type. In addition, mutant #24 had shown an increase of biomass accumulation approximately 1.65 fold compared with the wild type strain on day 5 that was entering early stationary phase. From these preliminary results, it could be feasible to identify D. salina mutants with significant improved growth rate, cell accumulation and biomass production compared to the wild type for the further study; this makes it possible to improve this microorganism as a platform for biotechnology application.

Keywords: Dunaliella salina, ethyl methyl sulfonate, growth rate, biomass

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Authors: Iulianna Taritsa, Kuldeep Neote, Eric Fossel

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Lysosome-induced immunogenic cell death (LIICD) is a powerful mechanism of targeting cancer cells that kills circulating malignant cells and primes the host’s immune cells against future remission. Current immunotherapies for cancer are limited in preventing recurrence – a gap that can be bridged by training the immune system to recognize cancer neoantigens. Lysosomal leakage can be induced therapeutically to traffic antigens from dying cells to dendritic cells, which can later present those tumorigenic antigens to T cells. Previous research has shown that oxidative agents administered in the tumor microenvironment can initiate LIICD. We generated a fusion protein between an oxidative agent known as xanthine oxidase (XO) and a mini-antibody specific for EGFR/HER2-sensitive breast tumor cells. The anti-EGFR single domain antibody fragment is uniquely sourced from llama, which is functional without the presence of a light chain. These llama micro-antibodies have been shown to be better able to penetrate tissues and have improved physicochemical stability as compared to traditional monoclonal antibodies. We demonstrate that the fusion protein created is stable and can induce early markers of immunogenic cell death in an in vitro human breast cancer cell line (SkBr3). Specifically, we measured overall cell death, as well as surface-expressed calreticulin, extracellular ATP release, and HMGB1 production. These markers are consensus indicators of ICD. Flow cytometry, luminescence assays, and ELISA were used respectively to quantify biomarker levels between treated versus untreated cells. We also included a positive control group of SkBr3 cells dosed with doxorubicin (a known inducer of LIICD) and a negative control dosed with cisplatin (a known inducer of cell death, but not of the immunogenic variety). We looked at each marker at various time points after cancer cells were treated with the XO/antibody fusion protein, doxorubicin, and cisplatin. Upregulated biomarkers after treatment with the fusion protein indicate an immunogenic response. We thus show the potential for this fusion protein to induce an anticancer effect paired with an adaptive immune response against EGFR/HER2+ cells. Our research in human cell lines here provides evidence for the success of the same therapeutic method for patients and serves as the gateway to developing a new treatment approach against breast cancer.

Keywords: apoptosis, breast cancer, immunogenic cell death, lysosome

Procedia PDF Downloads 198