Search results for: stem cell differentiation

Authors: Ishani Das, Avinash Padhi, Sitabja Mukherjee, Santosh Kar, Avinash Sonawane

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Activation of cell mediated and humoral immune responses to Mycobacterium tuberculosis (Mtb) are critical for protection. Herein, we show that mice immunized with Mtb lipid bound chitosan nanoparticles(NPs) induce secretion of prominent Th1 and Th2 cytokines in lymph node and spleen cells, and also induced significantly higher levels of IgG, IgG1, IgG2 and IgM in comparison to control mice measured by ELISA. Furthermore, significantly enhanced γδ-T cell activation was observed in lymph node cells isolated from mice immunized with Mtb lipid coated chitosan-NPs as compared to mice immunized with chitosan-NPs alone or Mtb lipid liposomes through flow cytometric analysis. Also, it was observed that in comparison to CD8+ cells, significantly higher CD4+ cells were present in both the lymph node and spleen cells isolated from mice immunized with Mtb lipid coated chitosan NP. In conclusion, this study represents a promising new strategy for efficient delivery of Mtb lipids using chitosan NPs to trigger enhanced cell mediated and antibody response against Mtb lipids.

Keywords: antibody response, chitosan nanoparticles, cytokines, mycobacterium tuberculosis lipids

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Authors: Vladimir S. Timofeev

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In this article the problem of distributional moments estimation is considered. The new approach of moments estimation based on usage of the characteristic function is proposed. By statistical simulation technique, author shows that new approach has some robust properties. For calculation of the derivatives of characteristic function there is used numerical differentiation. Obtained results confirmed that author’s idea has a certain working efficiency and it can be recommended for any statistical applications.

Keywords: characteristic function, distributional moments, robustness, outlier, statistical estimation problem, statistical simulation

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Authors: Mhuji Kilonzo, Patrick Ndakidemi, Musa Chacha

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Objective: To evaluate antibacterial activity from four selected medicinal plants namely Mystroxylon aethiopicum, Lonchocarpus capassa, Albizia anthelmentica and Myrica salicifolia used for management of bacterial infection in Tanzania. Methods: Minimum Inhibitory Concentration (MIC) of plants extracts against the tested bacterial species was determined by using 96 wells microdilution method. In this method, 50 μL of nutrient broth were loaded in each well followed by 50 μL of extract (100 mg/mL) to make a final volume of 100 μL. Subsequently, 50 μL were transferred from first rows of each well to the second rows and the process was repeated down the columns to the last wells from which 50 μL were discarded. Thereafter, 50 μL of the selected bacterial suspension were added to each well thus making a final volume of 100 μL. The lowest concentration which showed no bacterial growth was considered as MIC. Results: It was revealed that L. capassa leaf ethyl acetate extract exhibited antibacterial activity against Salmonella kisarawe and Salmonella typhi with MIC values of 0.39 and 0.781 mg/mL respectively. Likewise, L. capassa root bark ethyl acetate extracts inhibited growth of S. typhi and E. coli with MIC values of 0.39 and 0.781 mg/mL respectively. The M. aethiopicum leaf and root bark chloroform extracts displayed antibacterial activity against S. kisarawe and S. typhi respectively with MIC value of 0.781 mg/mL. The M. salicifolia stem bark ethyl acetate exhibited antibacterial activity against P. aeruginosa with MIC value of 0.39 mg/mL whereas the methanolic stem and root bark of the same plant inhibited the growth of Proteus mirabilis and Klebsiella pneumoniae with MIC value of 0.781 mg/mL. Conclusion: It was concluded that M. aethiopicum, L. capassa, A. anthelmentica and M. salicifolia are potential source of antibacterial agents. Further studies to establish structures of antibacterial and evaluate active ingredients are recommended.

Keywords: Albizia anthelmentica, Lonchocarpus capassa, Mystroxylon aethiopicum, Myrica salicifolia

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Authors: Mai M. Farid, Mona El-Shabrawy, Sameh R. Hussein, Ahmed Elkhateeb, El-Said S. Abdel-Hameed, Mona M. Marzouk

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Asphodelus aestivus Brot. Is a wild plant distributed in Egypt and is considered one of the five Asphodelus spp. from the family Asphodelaceae; it grows in dry grasslands and on rocky or sandy soil. The chemical components of A. aestivus flowers extract were analyzed using different chromatographic and spectral techniques and led to the isolation of two anthraquinones identified as emodin and emodin-O-glucoside. In addition to, five flavonoid compounds;kaempferol,Kaempferol-3-O-glucoside,Apigenin-6-C-glucoside-7-O-glucoside (Saponarine), luteolin 7-O-β-glucopyranoside, Isoorientin-O-malic acid which is a new compound in nature. The LC-ESI-MS/MS analysis of the flower extract of A. aestivus led to the identification of twenty- two compounds characterized by the presence of flavones, flavonols, and flavone C-glycosides. While GC/MS analysis led to the identification of 24 compounds comprising 98.32% of the oil, the major components of the oil were 9, 12, 15-Octadecatrieoic acid methyl ester 28.72%, and 9, 12-Octadecadieroic acid (Z, Z)-methyl ester 19.96%. In vitro cytotoxic activity of the aqueous methanol extract of A. aestivus flowers against HEPG2, HCT-116, MCF-7, and A549 culture was examined and showed moderate inhibition (62.3±1.1)% on HEPG2 cell line followed by (36.8±0.2)% inhibition on HCT-116 and a weak inhibition (5.7± 0.0.2) on MCF-7 cell line followed by (4.5± 0.4) % inhibition on A549 cell line and this is considered the first cytotoxic report of A. aestivus flowers.

Keywords: Anthraquinones, Asphodelus aestivus, Cytotoxic activity, Flavonoids, LC-ESI-MS/MS

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Authors: Hammad Khan, Wookeun Bae

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Major concerns regarding nitritation of low-strength ammonium wastewaters include low ammonium loading rates (usually below 0.2 kg/m3-d) and uncertainty about long-term stability of the process. The purpose of this study was to test a sequencing batch reactor (SBR) filled with cell-immobilized polyethylene glycol (PEG) pellets to see if it could achieve efficient and stable nitritation under various environmental conditions. SBR was fed with synthetic ammonium wastewater of 30±2 mg-N/L and pH: 8±0.05, maintaining the dissolved oxygen concentration of 1.7±0.2 mg/L and the temperature at 30±1oC. The reaction was easily converted to partial nitrification mode within a month by feeding relatively high ammonium substrate (~100 mg-N/L) in the beginning. We observed stable nitritation over 300 days with high ammonium loading rates (as high as ~1.1 kg-N/m3-d), nitrite accumulation rates (mostly over 97%) and ammonium removal rate (mostly over 95%). DO was a major limiting substrate when the DO concentration was below ~4 mg/L and the NH4+-N concentration was above 5 mg/L, giving almost linear increase in the ammonium oxidation rate with the bulk DO increase. Low temperatures mainly affected the reaction rate, which could be compensated for by increasing the pellet volume (i.e. biomass). Our results demonstrated that an SBR filled with small cell-immobilized PEG pellets could achieve very efficient and stable nitritation of a low-strength ammonium wastewater.

Keywords: ammonium loading rate (ALR), cell-immobilization, long-term nitritation, sequencing batch reactor (SBR), sewage treatment

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Authors: Ying Sun, Biao Cheng, Qing Lu, Xuefei Tao, Xiaoyu Lai, Cheng Guo, Dan Wang

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Fibrinogen-like protein 2 (FGL2) has recently been identified to play an important role in inflammatory diseases such as atherosclerosis through a thrombin-dependent manner. Here, a murine monoclonal antibody was raised against the critical residue Ser(89) of FGL2, and the effects of the anti-FGL2 mAb (SPF89) were analyzed in human umbilical vein endothelial cells (HUVECs) and THP-1 cells. Firstly, it was proved that SPF89, which belongs to the IgG1 subtype with a KD value of 44.5 pM, could specifically show the expression levels of protein FGL2 in different cell lines of known target gene status. The lipopolysaccharide (LPS)-mediated endothelial cell proliferation was significantly inhibited with a decline of phosphorylation nuclear factor-κB (NF-κB) in a dose-dependent manner after SPF89 treatment. Furthermore, SPF89 reduced LPS-induced expression of adhesion molecules and inflammatory cytokines such as vascular cell adhesion molecule-1, tumor necrosis factor-α, Matrix metalloproteinase MMP-2, Integrin αvβ3, and interleukin-6 in HUVECs. In macrophage-like THP-1 cells, SPF89 effectively inhibited LPS and low-density lipoprotein-induced foam cell formation. However, these anti-inflammatory and anti-atherosclerotic effects of anti-FGL2 mAb in HUVECs and THP-1 cells were significantly reduced after treatment with an NF-κB inhibitor PDTC. All the above suggest, by efficiently inhibiting LPS-induced pro-inflammatory effects in vascular endothelial cells by attenuating NF-κB dependent pathway, the new anti-FGL2 mAb SPF89 could to be a potential therapeutic candidate for protecting the vascular endothelium against inflammatory diseases such as atherosclerosis. This work was supported by the Program of Sichuan Science and Technology Department (2017FZ0069) and Collaborative Innovation Program of Sichuan for Elderly Care and Health(YLZBZ1511).

Keywords: monoclonal antibody, fibrinogen like protein 2, inflammation, endothelial cells

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Authors: Shen Jian, Hu Jie, Zhu Guo Niu, Peng Ying Hong

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There are two issues exited in the traditional visual prosthesis: lacking systematic method and the low level of humanization. To tackcle those obstacles, a visual prosthesis design method based on biologically inspired design is proposed. Firstly, a constrained FBS knowledge cell model is applied to construct the functional model of visual prosthesis in biological field. Then the clustering results of engineering domain are ob-tained with the use of the cross-domain knowledge cell clustering algorithm. Finally, a prototype system is designed to support the bio-logically inspired design where the conflict is digested by TRIZ and other tools, and the validity of the method is verified by the solution scheme

Keywords: knowledge-based engineering, visual prosthesis, biologically inspired design, biomedical engineering

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Authors: Yik Sin Chan, Nam Weng Sit, Fook Yee Chye, van Ofwegen Leen, de Voogd Nicole, Kong Soo Khoo

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Chikungunya fever is an arboviral disease transmitted by the Aedes mosquitoes. It has resulted in epidemics of the disease in tropical countries in the Indian Ocean and South East Asian regions. The recent spread of this disease to the temperate countries such as France and Italy, coupled with the absence of vaccines and effective antiviral drugs make chikungunya fever a worldwide health threat. This study aims to investigate the anti-chikungunya virus activity of selected marine organism samples collected from Malaysian coastal areas, including seaweeds (Caulerpa racemosa, Caulerpa sertularioides and Kappaphycus alvarezii), a soft coral (Lobophytum microlobulatum) and a sponge (Spheciospongia vagabunda). Following lyophilization (oven drying at 40C for K. alvarezii) and grinding to powder form, each sample was subjected to sequential solvent extraction using hexane, chloroform, ethyl acetate, ethanol, methanol and distilled water in order to extract bioactive compounds. The antiviral activity was evaluated using monkey kidney epithelial (Vero) cells infected with the virus (multiplicity of infection=1). The cell viability was determined by Neutral Red uptake assay. 70% of the 30 extracts showed weak inhibitory activity with cell viability ≤30%. Seven of the extracts exhibited moderate inhibitory activity (cell viability: 31%-69%). These were the chloroform, ethyl acetate, ethanol and methanol extracts of C. racemosa; chloroform and ethyl acetate extracts of L. microlobulatum; and the chloroform extract of C. sertularioides. Only the hexane and ethanol extracts of L. microlobulatum showed strong inhibitory activity against the virus, resulting in cell viabilities (mean±SD; n=3) of 73.3±2.6% and 79.2±0.9%, respectively. The corresponding mean 50% effective concentrations (EC50) for the extracts were 14.2±0.2 and 115.3±1.2 µg/mL, respectively. The ethanol extract of the soft coral L. microlobulatum appears to hold the most promise for further characterization of active principles as it possessed greater selectivity index (SI>5.6) compared to the hexane extract (SI=2.1).

Keywords: antiviral, seaweed, sponge, soft coral, vero cell

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Authors: Cuili Yang, Chengcao Sun, Ruilin Xue, Yongyong Xi, Liang Wang, Dejia Li

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Backgrounds: Previous studies showed that Sirt1 plays an important role in C2C12 myoblast cell proliferation, but the mechanism(s) involved in this process remains unclear. This work was undertaken to determine if Myostatin participates in the regulation of C2C12 proliferation by Sirt1. Methods: We administrated the Sirt1 activator resveratrol, inhibitor Nicotinamide (NAM) and Myostatin inhibitor SB431542 on C2C12 myoblast cells. Cell viability was evaluated by CCK8 assay. The expression of Sirt1 and MyoD were detected by qRT-PCR. Utilizing western blot to determinate the expression of myostatin, P107 and p-P107. Results: Our results showed that resveratrol promoted the proliferation of C2C12 myoblast cells, while NAM suppressed the proliferation of C2C12 myoblast cells; SB431542 promoted the proliferation of C2C12 myoblast cells and attenuated the inhibition effect of NAM on C2C12 myoblast cells proliferation; Resveratrol can significantly increase the expression of Sirt1 and MyoD, decrease the expression of Myostatin, while NAM can significantly down-regulate the expression of Sirt1, MyoD and the phosphorylation of P107(p-P107), but up-regulate the expression of Myostatin and the protein P107; SB431542 can significantly mitigate the effect of NAM on the expression of MyoD, P107, and p-P107. Conclusions: Taken together, these results indicate that Sirt1 promotes the proliferation of C2C12 myoblast cells via Myostatin signaling pathway.

Keywords: Sirt1, C2C12 cells, proliferation, myostatin signaling pathway

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Authors: Kohei Inoue, Kenji Hara, Kiichi Urahama

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We describe a relationship between integral images and differential images. First, we derive a simple difference filter from conventional integral image. In the derivation, we show that an integral image and the corresponding differential image are related to each other by simultaneous linear equations, where the numbers of unknowns and equations are the same, and therefore, we can execute the integration and differentiation by solving the simultaneous equations. We applied the relationship to an image fusion problem, and experimentally verified the effectiveness of the proposed method.

Keywords: integral images, differential images, differential filters, image fusion

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Authors: Samarjeet Singh, Shubhank Arya, Shubham Chauhan

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In recent years, the rise in carbon emissions and the widespread effects of global warming have brought new energy vehicles into the spotlight. Electric vehicles (EVs) and hydrogen fuel cell vehicles (HFCVs), both producing zero tailpipe emissions, are seen as promising alternatives. This paper examines the working, structural characteristics, and safety designs of EVs and HFCVs, comparing their carbon emissions, charging infrastructure, energy efficiency, and safety features. The analysis reveals that both EVs and HFCVs significantly reduce carbon emissions and enhance safety compared to traditional vehicles, with EVs showing greater emission reductions. Moreover, EVs are advancing more rapidly in terms of charging infrastructure compared to hydrogen energy vehicles. However, HFCVs exhibit lower energy efficiency than EVs. In terms of safety, both types surpass conventional vehicles, though EVs are more prone to overheating and fire hazards due to battery design issues. Current research suggests that EV technology and its supporting infrastructure are more comprehensive, cost-effective, and efficient in reducing carbon emissions. With continued investment in the development of new energy vehicles and potential advancements in hydrogen energy production, the future for HFCVs appears promising. The paper also expresses optimism for innovative solutions that could accelerate the growth of hydrogen energy vehicles.

Keywords: electric vehicles, fuel cell electric vehicles, automotive engineering, energy transition

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Authors: Mahdiye Khorasani, Arjun Vijay, Ali Mashayekh, Christian Trapp

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The performance of Proton Exchange Membrane Fuel Cell (PEMFC) is highly dependent on the pressure and mass flow of media (Hydrogen and air) throughout the cells and the stack. Higher pressure, on the one hand, results in higher output power of the stack but, on the other hand, increases the electrical power demand of the compressor. In this work, a simulation model of a PEMFC system for vehicle and airplane applications is developed. With this new model, the effect of different pressures and air mass flow rates are investigated to discover the optimum operating point in a PEMFC system, and innovative operation strategies are implemented to optimize reactants flow while minimizing electrical power demand of the compressor for optimum performance. Additionally, a fuel cell system test bench is set up, which contains not only all the auxiliary components for conditioning the gases, reactants, and flows but also a dynamic titling table for testing different orientations of the stack to simulate the flight conditions during take-off and landing and off-road-vehicle scenarios. The results of simulation will be tested and validated on the test bench for future works.

Keywords: air mass flow effect, optimization of operation, pressure effect, PEMFC system, PEMFC system simulation

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Authors: Ripon Sarkar, Kabita Chaterjee, Ananya Barui

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Smoking is one of the leading causes of oral cancer. Cigarette smoke affects various cellular parameters and alters molecular metabolism of cells. Epithelial cells losses their cytoskeleton structure, membrane integrity, cellular polarity that subsequently initiates the process of epithelial cells to mesenchymal transition due to long exposure of cigarette smoking. It changes the normal cellular metabolic activity which induces oxidative stress and enhances the reactive oxygen spices (ROS) formation. Excessive ROS and associated oxidative stress are considered to be a driving force in alteration in cellular phenotypes, polarity distribution and mitochondrial metabolism. Noninvasive assessment of such parameters plays essential role in development of routine screening system for early diagnosis of oral cancer. Electrical cell-substrate impedance sensing (ECIS) is one of such method applied for detection of cellular membrane impedance which can be correlated to cell membrane integrity. Present study intends to explore the alteration in cellular impedance along with the expression of cellular polarity molecules and cytoskeleton distributions in oral epithelial cells of habitual smokers and to correlate the outcome to that of clinically diagnosed oral leukoplakia and oral squamous cell carcinoma patients. Total 80 subjects were categorized into four study groups: nonsmoker (NS), cigarette smoker (CS), oral leukoplakia (OLPK) and oral squamous cell carcinoma (OSCC). Cytoskeleton distribution was analyzed by staining of actin filament and generation of ROS was measured using assay kit using standard protocol. Cell impedance was measured through ECIS method at different frequencies. Expression of E-cadherin and protease-activated receptor (PAR) proteins were observed through immune-fluorescence method. Distribution of actin filament is well organized in NS group however; distribution pattern was grossly varied in CS, OLPK and OSCC. Generation of ROS was low in NS which subsequently increased towards OSCC. Expressions of E-cadherin and change in cellular electrical impedance in different study groups indicated the hallmark of cancer progression from NS to OSCC. Expressions of E-cadherin, PAR protein, and cell impedance were decreased from NS to CS and farther OSCC. Generally, the oral epithelial cells exhibit apico-basal polarity however with cancer progression these cells lose their characteristic polarity distribution. In this study expression of polarity molecule and ECIS observation indicates such altered pattern of polarity among smoker group. Overall the present study monitored the alterations in intracellular ROS generation and cell metabolic function, membrane integrity in oral epithelial cells in cigarette smokers. Present study thus has clinical significance, and it may help in developing a noninvasive technique for early diagnosis of oral cancer amongst susceptible individuals.

Keywords: cigarette smoking, early oral cancer detection, electric cell-substrate impedance sensing, noninvasive screening

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Authors: Shaimaa Eltanani, Thangal Yumnamcha, Ahmed S. Ibrahim

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Purpose: Mitochondria dysfunction is central to breaking the barrier integrity of retinal endothelial cells (RECs) in various blinding eye diseases such as diabetic retinopathy and retinopathy of prematurity. Therefore, we aimed to dissect the role of different mitochondrial components, specifically, those of oxidative phosphorylation (OxPhos), in maintaining the barrier function of RECs. Methods: Electric cell-substrate impedance sensing (ECIS) technology was used to assess in real-time the role of different mitochondrial components in the total impedance (Z) of human RECs (HRECs) and its components; the capacitance (C) and the total resistance (R). HRECs were treated with specific mitochondrial inhibitors that target different steps in OxPhos: Rotenone for complex I; Oligomycin for ATP synthase; and FCCP for uncoupling OxPhos. Furthermore, data were modeled to investigate the effects of these inhibitors on the three parameters that govern the total resistance of cells: cell-cell interactions (Rb), cell-matrix interactions (α), and cell membrane permeability (Cm). Results: Rotenone (1 µM) produced the greatest reduction in the Z, followed by FCCP (1 µM), whereas no reduction in the Z was observed after the treatment with Oligomycin (1 µM). Following this further, we deconvoluted the effect of these inhibitors on Rb, α, and Cm. Firstly, rotenone (1 µM) completely abolished the resistance contribution of Rb, as the Rb became zero immediately after the treatment. Secondly, FCCP (1 µM) eliminated the resistance contribution of Rb only after 2.5 hours and increased Cm without considerable effect on α. Lastly, Oligomycin had the lowest impact among these inhibitors on Rb, which became similar to the control group at the end of the experiment without noticeable effects on Cm or α. Conclusion: These results demonstrate differential roles for complex I, complex V, and coupling of OxPhos in maintaining the barrier functionality of HRECs, in which complex I being the most important component in regulating the barrier functionality and the spreading behavior of HRECs. Such differences can be used in investigating gene expression as well as for screening selective agents that improve the functionality of complex I to be used in the therapeutic approach for treating REC-related retinal diseases.

Keywords: human retinal endothelial cells (hrecs), rotenone, oligomycin, fccp, oxidative phosphorylation, oxphos, capacitance, impedance, ecis modeling, rb resistance, α resistance, and barrier integrity

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Authors: N. Ojaroudi Parchin, H. Jahanbakhsh Basherlou, Y. Al-Yasir, A. M. Abdulkhaleq, R. A. Abd-Alhameed, P. S. Excell

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A compact design of multi-sector patch antenna array for 60 GHz applications is presented and discussed in details. The proposed design combines five 1×8 linear patch antenna arrays, referred to as sectors, in a multi-sector configuration. The coaxial-fed radiation elements of the multi-sector array are designed on 0.2 mm Rogers RT5880 dielectrics. The array operates in the frequency range of 58-62 GHz and provides switchable directional/omnidirectional radiation beams with high gain and high directivity characteristics. The designed multi-sector array exhibits good performances and could be used in the fifth generation (5G) cellular networks.

Keywords: mm-wave communications, multi-sector array, patch antenna, small cell networks

Procedia PDF Downloads 157

Authors: Yunjia Yang, Peng Li, Gordon Xu, Timothy Mahony, Bing Zhang, Neena Mitter, Karishma Mody

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Sheep flystrike is one of the most economically important diseases affecting the Australian sheep and wool industry (>356M/annually). Currently, control of Lucillia cuprina relies almost exclusively on chemicals controls, and the parasite has developed resistance to nearly all control chemicals used in the past. It is, therefore, critical to develop an alternative solution for the sustainable control and management of flystrike. RNA interference (RNAi) technologies have been successfully explored in multiple animal industries for developing parasites controls. This research project aims to develop a RNAi based biological control for sheep blowfly. Double-stranded RNA (dsRNA) has already proven successful against viruses, fungi, and insects. However, the environmental instability of dsRNA is a major bottleneck for successful RNAi. Bentonite polymer (BenPol) technology can overcome this problem, as it can be tuned for the controlled release of dsRNA in the gut challenging pH environment of the blowfly larvae, prolonging its exposure time to and uptake by target cells. To investigate the potential of BenPol technology for dsRNA delivery, four different BenPol carriers were tested for their dsRNA loading capabilities, and three of them were found to be capable of affording dsRNA stability under multiple temperatures (4°C, 22°C, 40°C, 55°C) in sheep serum. Based on stability results, dsRNA from potential targeted genes was loaded onto BenPol carriers and tested in larvae feeding assays, three genes resulting in knockdowns. Meanwhile, a primary blowfly embryo cell line (BFEC) derived from L. cuprina embryos was successfully established, aim for an effective insect cell model for testing RNAi efficacy for preliminary assessments and screening. The results of this study establish that the dsRNA is stable when loaded on BenPol particles, unlike naked dsRNA rapidly degraded in sheep serum. The stable nanoparticle delivery system offered by BenPol technology can protect and increase the inherent stability of dsRNA molecules at higher temperatures in a complex biological fluid like serum, providing promise for its future use in enhancing animal protection.

Keywords: lucilia cuprina, primary cell line establishment, RNA interference, insect cell transfection

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Authors: P. Praveen Kumar, P. Vimal Prabhu, Renu John

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In biology most microscopy specimens, in particular living cells are transparent. In cell imaging, it is hard to create an image of a cell which is transparent with a very small refractive index change with respect to the surrounding media. Various techniques like addition of staining and contrast agents, markers have been applied in the past for creating contrast. Many of the staining agents or markers are not applicable to live cell imaging as they are toxic. In this paper, we report theoretical and experimental results from quantitative phase imaging of yeast cells with a commercial bright field microscope. We reconstruct the phase of cells non-interferometrically based on the transport of intensity equations (TIE). This technique estimates the axial derivative from positive through-focus intensity measurements. This technique allows phase imaging using a regular microscope with white light illumination. We demonstrate nano-metric depth sensitivity in imaging live yeast cells using this technique. Experimental results will be shown in the paper demonstrating the capability of the technique in 3-D volume estimation of living cells. This real-time imaging technique would be highly promising in real-time digital pathology applications, screening of pathogens and staging of diseases like malaria as it does not need any pre-processing of samples.

Keywords: axial derivative, non-interferometric imaging, quantitative phase imaging, transport of intensity equation

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Authors: M. Y. Abubakar, Ipinjolu J. K., Yuzine B. Esa, Magawata I., Hassan W. A., Turaki A. A.

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Catfish (Heterobranchus spp.) is a major freshwater fish that are widely distributed in Nigeria waters and are gaining rapid aquaculture expansion. However, indiscriminate artificial crossbreeding of the species with others poses a threat to their biodiversity. There is a paucity of information about the genetic variability, hence this insight on the genetic variability is badly needed, not only for the species conservation but for aquaculture expansion. In this study, we tested the level of Genetic diversity, population differentiation and phylogenetic relationship analysis on 35 individuals of two populations of Heterobranchus bidorsalis and 29 individuals of three populations of Heterobranchus longifilis using the mitochondrial cytochrome c oxidase subunit I (mtDNA COI) gene sequence. Nucleotide sequences of 650 bp fragment of the COI gene of the two species were compared. In the whole 4 and 5 haplotypes were distinguished in the populations of H. bidorsalis & H. longifilis with accession numbers (MG334168 - MG334171 & MG334172 to MG334176) respectively. Haplotypes diversity indices revealed a range of 0.59 ± 0.08 to 0.57 ± 0.09 in H. bidorsalis and 0.000 to 0.001051 ± 0.000945 in H. longifilis population, respectively. Analysis of molecular variance (AMOVA) revealed no significant variation among H. bidorsalis population of the Niger & Benue Rivers, detected significant genetic variation was between the Rivers of Niger, Kaduna and Benue population of H. longifilis. Two main clades were recovered, showing a clear separation between H. bidorsalis and H. longifilis in the phylogenetic tree. The mtDNA COI genes studied revealed high gene flow between populations with no distinct genetic differentiation between the populations as measured by the fixation index (FST) statistic. However, a proportion of population-specific haplotypes was observed in the two species studied, suggesting a substantial degree of genetic distinctiveness for each of the population investigated. These findings present the description of the species character and accessions of the fish’s genetic resources, through gene sequence submitted in Genetic database. The data will help to protect their valuable wild resource and contribute to their recovery and selective breeding in Nigeria.

Keywords: AMOVA, genetic diversity, Heterobranchus spp., mtDNA COI, phylogenetic tree

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Authors: Bhumika Wadhwa, Fayaz Malik

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The serine/threonine protein kinase B (PKB) also known as Akt, is one of the multifaceted kinase in human kinome, existing in three isoforms. Akt plays a vital role in phosphoinositide 3-kinase (PI3K) mediated oncogenesis in various malignancies and is one of the attractive targets for cancer drug discovery. The functional significance of an individual isoform of Akt is not redundant in cancer cell proliferation and metastasis instead Akt isoforms play distinct roles during metastasis; thereby regulating EMT. This study aims to determine isoform specific functions of Akt in cancer. The results obtained suggest that Akt1 restrict tumor invasion, whereas Akt2 promotes cell migration and invasion by various techniques like MTT, wound healing and invasion assay. Similarly, qRT-PCR also revealed that Akt3 has shown promising results in promoting cancer cell migration. Contrary to pro-oncogenic properties attributed to Akt, it is to be understood how various isoforms of Akt compensates each other in the regulation of common pathways during cancer progression and drug resistance. In conclusion, this study aims to target selective isoforms which is essential to inhibit cancer. However, the question now is whether, and how much, Akt inhibition will be tolerated in the clinic remains to be answered and the experiments will have to address the question of which combinations of newly devised Akt isoform specific inhibitors exert a favourable therapeutic effect in in vivo models of cancer to provide the therapeutic window with minimal toxicity.

Keywords: Akt isoforms, cancer, drug resistance, epithelial mesenchymal transition

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Authors: S. H. Jang

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Posterior reversible encephalopathy syndrome (PRES) is characterized by acute neurologic symptoms (visual loss, headache, altered mentality and seizures) and by typical imaging findings (bilateral subcortical and cortical edema with predominatly posterior distribution). Nephrotic syndrome is a syndrome comprising signs of proteinuria, hypoalbuminemia, and edema. It is well known that hypertension predispose patient with nephrotic syndrome to PRES. A 45-year old male was referred for suddenly developed vertigo, disequilibrium. He had previous history of nephrotic syndrome. His medical history included diabetes controlled with medication. He was hospitalized because of generalized edema a few days ago. His vital signs were stable. On neurologic examination, his mental state was alert. Horizontal nystagmus to right side on return to primary position was observed. He showed good grade motor weakness and ataxia in right upper and lower limbs without other sensory abnormality. Brain MRI showed increased signal intensity in FLAIR image, decreased signal intensity in T1 image and focal enhanced lesion in T1 contrast image at whole midbrain, pons and cerebellar peduncle symmetrically, which was compatible with vasogenic edema. Laboratory findings showed severe proteinuria and hypoalbuminemia. He was given intravenous dexamethasone and diuretics to reduce vasogenic edema and raise the intra-vascular osmotic pressure. Nystagmus, motor weakness and limb ataxia improved gradually over 2 weeks; He recovered without any neurologic symptom and sign. Follow-up MRI showed decreased vasogenic edema fairly. We report a case of brain stem PRES in normotensive, nephrotic syndrome patient.

Keywords: posterior reversible encephalopathy syndrome, MRI, nephrotic syndrome, vasogenic brain edema

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Authors: Livinus A. Obasi, Augustine N. Ajah

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Biotechnology in recent times has tried to develop a mechanism whereby sustainable electricity can be generated by the activity of microorganisms on waste and renewable biomass (often regarded as “negative value”) in a device called microbial fuel cell, MFC. In this paper, we established how the biocatalytic activities of bacteria on organic matter (substrates) produced some electrons with the associated removal of some water pollution parameters; Biochemical oxygen demand (BOD), chemical oxygen demand (COD) to the tune of 77.2% and 88.3% respectively from a petrochemical sanitary wastewater. The electricity generation was possible by conditioning the bacteria to operate anaerobically in one chamber referred to as the anode while the electrons are transferred to the fully aerated counter chamber containing the cathode. Power densities ranging from 12.83 mW/m² to 966.66 mW/m² were achieved using a dual-chamber starch membrane MFC experimental set-up. The maximum power density obtained in this research shows an improvement in the use of low cost MFC set up to achieve power production. Also, the level of organic matter removal from the sanitary waste water by the operation of this device clearly demonstrates its potential benefit in achieving an improved benign environment. The beauty of the MFCs is their potential utility in areas lacking electrical infrastructures like in most developing countries.

Keywords: bioelectricity, COD, microbial fuel cell, sanitary wastewater, wheat starch

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Authors: Osama Terra, Hatem Hussein, Adriaan Van Brakel

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Dense wavelength division multiplexing (DWDM) technology requires tight specification and therefore measurement of wavelength accuracy and stability of the telecommunication lasers. Thus, calibration of the used Optical Spectrum Analyzers (OSAs) that are used to measure wavelength is of a great importance. Proficiency testing must be performed on such measuring activity to insure the accuracy of the measurement results. In this paper, a new comparison scheme is introduced to test the performance of such calibrations. This comparison scheme is implemented between NIS-Egypt and NMISA-South Africa for the calibration of the wavelength scale of an OSA. Both institutes employ reference gas cell to calibrate OSA according to the standard IEC/ BS EN 62129 (2006). The result of this comparison is compiled in this paper.

Keywords: OSA calibration, HCN gas cell, DWDM technology, wavelength measurement

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Authors: B. Niez, B. Moulia, J. Dlouha, E. Badel

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Trees adjust their development to the environmental conditions they experience. Storms events of last decades showed that acclimation of trees to mechanical stresses due to wind is a very important process that allows the trees to sustain for long years. In the future, trees will experience new wind patterns, namely, more often strong winds and fewer daily moderate winds. Moreover, these patterns will go along with drought periods that may interact with the capacity of trees to adjust their growth to mechanical stresses due to wind. It is necessary to understand the mechanisms of wood functional acclimations to environmental conditions in order to predict their behaviour and in order to give foresters and breeders the relevant tools to adapt their forest management. This work aims to study how trees adjust the mechanical and hydraulic functions of their wood to environmental stresses and how this acclimation may be beneficial for the tree to resist to future stresses. In this work, young poplars were grown under controlled climatic conditions that include permanent environmental stress (daily mechanical stress of the stem by bending and/or hydric stress). Then, the properties of wood formed under these stressed conditions were characterized. First, hydraulic conductivity and sensibility to cavitation were measured at the tissue level in order to evaluate the changes in water transport capacity. Secondly, bending tests and Charpy impact tests were carried out at the millimetric scale to locally measure mechanical parameters such as elastic modulus, elastic limit or rupture energy. These experimental data allow evaluating the impacts of mechanical and water stress on the wood material. At the stem level, they will be merged in an integrative model in order to evaluate the beneficial aspect of wood acclimation for trees.

Keywords: acclimation, environmental stresses, hydraulics, mechanics, wood

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Authors: Johanna Perdomo, Riki Lamont, Edmund Pickering, Naomi C. Paxton, Maria A. Woodruff

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Background: Tissue Engineering (TE) has combined advanced materials, such as biomaterials, to create affordable scaffolds and dynamic systems to generate stimulation of seeded cells on these scaffolds, improving and maintaining the cellular growth process in a cell culture. However, Few TE skin products have been clinically translated, and more research is required to produce highly biomimetic skin substitutes that mimic the native elasticity of skin in a controlled manner. Therefore, this work will be focused on the fabrication of a novel mechanical system to enhance the TE treatment approaches for the reparation of damaged tissue skin. Aims: To archive this, a soft robotic device will be created to emulate different deformation of skin stress. The design of this soft robot will allow the attachment of scaffolds, which will then be mechanically actuated. This will provide a novel and highly adaptable platform for dynamic cell culture. Methods: Novel, low-cost soft robot is fabricated via 3D printed moulds and silicone. A low cost, electro-mechanical device was constructed to actuate the soft robot through the controlled combination of positive and negative air pressure to control the different state of movements. Mechanical tests were conducted to assess the performance and calibration of each electronic component. Similarly, pressure-displacement test was performed on scaffolds, which were attached to the soft robot, applying various mechanical loading regimes. Lastly, digital image correlation test was performed to obtain strain distributions over the soft robot’s surface. Results: The control system can control and stabilise positive pressure changes for long hours. Similarly, pressure-displacement test demonstrated that scaffolds with 5µm of diameter and wavy geometry can displace at 100%, applying a maximum pressure of 1.5 PSI. Lastly, during the inflation state, the displacement of silicone was measured using DIC method, and this showed a parameter of 4.78 mm and strain of 0.0652. Discussion And Conclusion: The developed soft robot system provides a novel and low-cost platform for the dynamic actuation of tissue scaffolds with a target towards dynamic cell culture.

Keywords: soft robot, tissue engineering, mechanical stimulation, dynamic cell culture, bioreactor

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Authors: Dileep K. Verma, Sunil K. Lal

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Influenza still remains one of the most challenging diseases posing significant threat to public health causing seasonal epidemics and pandemics. Previous studies suggest that influenza hemagglutinin is essential for viral attachment to host sialic acid receptors and concentrate in lipid rafts for efficient viral fusion. Studies also reported selective nature of Influenza virus to utilize rafts micro-domain for efficient virus assembly and budding. However, the detailed mechanism of Influenza A Virus (IAV) binding to host cell membrane and entry inside the host remains elusive. In the present study, we investigated if host membrane lipid rafts play any significant role in early life cycle events of influenza A virus. Role of host lipid rafts was studied using raft disruption method by extraction of cholesterol and Methyl-β-Cyclodextrin was used to remove membrane cholesterol. We observed co-localization of Influenza A Virus to lipid rafts by visualization of known lipid raft marker GM1 on host cell membrane. Co-localization suggest direct involvement of these micro-domain in initiation of IAV life cycle. We found significant reduction in influenza A virus adsorption in raft disrupted target host cells indicating poor binding and attachment in absence of coherent membrane rafts. Taken together, the results of present study provide evidence for critical involvement of host lipid rafts and its constituents in adsorption process of Influenza A Virus and suggests crucial involvement in other early events of IAV life cycle. The present study opens a new domain to study influenza virus-host interaction and to combat flu at the very early steps of viral life cycle.

Keywords: lipid raft, adsorption, cholesterol, methyl-β-cyclodextrin, GM1

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Authors: Husham Bayazed

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Purpose of Presentation: The eye is one of a few sites in the body with immune privilege (IP). However, this IP is relatively easily bypassed in the face of sufficient strong local or systemic immunological responses. As immune responses are crucial elements of the repair response, the eye has developed distinct mechanisms to deliver immune responses to injury in the avascular regions of the eye. This presentation may cover and provide an overview of the mechanisms that dictate immune cell trafficking to the local ocular microenvironment in response to different autoimmune and inflammatory-mediated diseases. Recent Findings: Literature reviews declare that immune responses and inflammation play a key role in a diverse range of eye diseases. In recent years, our understanding of ocular immune responses has widely spread in ocular surface inflammation, uveitis, age-related macular degeneration (AMD), glaucoma, transplantation rejection, and other ocular diseases. It is becoming increasingly clear that multiple seemingly unrelated diseases involve immune responses with common themes in their ocular pathogenesis. Recent studies are focusing on elucidating the pathogenesis of ocular inflammatory disease to identify new targets for immunotherapy that will not only improve efficacy but also minimize adverse effects from traditional therapy. Summary: While IP was believed to protect the eye from day-to-day inflammatory insults, however, it is relatively easily breached in the face of different strong local or systemic immunological and inflammatory responses. Therefore, the ocular immune response encapsulates the full range of classical and non-classical immune responses and demonstrates many features which are reflected in other tissues, but eye tissues, by modifying these responses, may reveal unexpected and novel findings which are relevant to immune responses generally. This may have therapeutic potential for new targeting immunotherapy, restoring immune tolerance in ocular autoimmune and inflammatory diseases, and preventing rejection such as stem cells, currently being considered for treatment of worldwide blinding diseases such as AMD.

Keywords: ocular diseases, immunology, immune privilege, immunotherapy

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Authors: Katarína Čurová, Leonard Siegfried, Radka Vargová, Marta Kmeťová, Vladimír Hrabovský

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Introduction: Cytolethal distending toxins (CDTs) represent intracellular acting proteins which interfere with cell cycle of eukaryotic cells. They are produced by Gram-negative bacteria with afinity to mucocutaneous surfaces and could play a role in the pathogenesis of various diseases. CDTs induce DNA damage probably through DNAse activity, which causes cell cycle arrest and leads to further changes (cell distension and death, apoptosis) depending on the cell type. Five subtypes of CDT (I to V) were reported in E. coli. Methods: We examined 252 E. coli strains belonging to four different groups. Of these strains, 57 were isolated from patients with diarrhea, 65 from patients with urinary tract infections (UTI), 65 from patients with sepsis and 65 from patients with other extraintestinal infections (mostly surgical wounds, decubitus ulcers and respiratory tract infections). Identification of these strains was performed by MALDI-TOF analysis and detection of genes encoding CDTs and determination of the phylogenetic group was performed by PCR. Results: In this study, we detected presence of cdt genes in 11 of 252 E. coli strains tested (4,4 %). Four cdt positive E. coli strains were confirmed in group of UTI (6,15 %), three cdt positive E. coli strains in groups of diarrhea (5,3 %) and other extraintestinal infections (4,6 %). The lowest incidence, one cdt positive E. coli strain, was observed in group of sepsis (1,5 %). All cdt positive E. coli strains belonged to phylogenetic group B2. Conclusion: CDT-producing E. coli are isolated in a low percentage from patients with intestinal and extraintestinal infections, including sepsis and our results correspond with these studies. A weak prevalence of cdt genes suggests that CDTs are not major virulence factors but in combination with other virulence factors may increase virulence potential of E. coli. We suppose that all 11 cdt positive E. coli strains represent real pathogens because they belong to the phylogenetic group B2 which is pathogenic lineage for bacteria E. coli.

Keywords: cytolethal distending toxin, E. coli, phylogenetic group, extraintestinal infection, diarrhea

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Authors: Zemlianskaya Daria, Egor Stadnichuk, Ekaterina Svechnikova

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Relativistic runaway electron avalanches (RREAs) are generally accepted as a source of thunderstorms gamma-ray radiation. Avalanches' dynamics in the electric fields can lead to their multiplication via gamma-rays and positrons, which is called relativistic feedback. This report shows that a non-uniform electric field geometry leads to the new RREAs multiplication mechanism - “geometric feedback”, which occurs due to the exchange of high-energy particles between different accelerating regions within a thundercloud. This report will present the results of the simulation in GEANT4 of feedback in a spherical cell. Necessary conditions for the occurrence of geometric feedback were obtained from it.

Keywords: electric field, GEANT4, gamma-rays, relativistic runaway electron avalanches (RREAs), relativistic feedback, the thundercloud

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Authors: Pujari Muruga, T. K. Radhakrishnan, N. Samsudeen

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Hydrogen is considered as the most important energy carrier and fuel of the future because of its high energy density and zero emission properties. Microbial Electrolysis Cell (MEC) is a new and promising approach for hydrogen production from organic matter, including wastewater and other renewable resources. By utilizing anode microorganism activity, MEC can produce hydrogen gas with smaller voltages (as low as 0.2 V) than those required for electrolytic hydrogen production ( ≥ 1.23 V). The hydrogen production processes of the MEC reactor are very nonlinear and highly complex because of the presence of microbial interactions and highly complex phenomena in the system. Increasing the hydrogen production rate and lowering the energy input are two important challenges of MEC technology. The mathematical model of the MEC is based on material balance with the integration of bioelectrochemical reactions. The main objective of the research is to produce biohydrogen by selecting the optimum current and controlling applied voltage to the MEC. Precise control is required for the MEC reactor, so that the amount of current required to produce hydrogen gas can be controlled according to the composition of the substrate in the reactor. Various simulation tests involving multiple set-point changes disturbance and noise rejection were performed to evaluate the performance using PID controller tuned with Ziegler Nichols settings. Simulation results shows that other good controller can provide better control effect on the MEC system, so that higher hydrogen production can be obtained.

Keywords: microbial electrolysis cell, hydrogen production, applied voltage, PID controller

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Authors: I-Ling Chang, Jer-An Chen

Abstract:

The elastic properties and fracture of two-dimensional graphene were calculated purely from the atomic bonding (stretching and bending) based on molecular mechanics method. Considering the representative unit cell of graphene under various loading conditions, the deformations of carbon bonds and the variations of the interlayer distance could be realized numerically under the geometry constraints and minimum energy assumption. In elastic region, it was found that graphene was in-plane isotropic. Meanwhile, the in-plane deformation of the representative unit cell is not uniform along armchair direction due to the discrete and non-uniform distributions of the atoms. The fracture of graphene could be predicted using fracture criteria based on the critical bond length, over which the bond would break. It was noticed that the fracture behavior were directional dependent, which was consistent with molecular dynamics simulation results.

Keywords: energy minimization, fracture, graphene, molecular mechanics

Procedia PDF Downloads 402