Search results for: particle per cell
3898 A Novel Epitope Prediction for Vaccine Designing against Ebola Viral Envelope Proteins
Authors: Manju Kanu, Subrata Sinha, Surabhi Johari
Abstract:
Viral proteins of Ebola viruses belong to one of the best studied viruses; however no effective prevention against EBOV has been developed. Epitope-based vaccines provide a new strategy for prophylactic and therapeutic application of pathogen-specific immunity. A critical requirement of this strategy is the identification and selection of T-cell epitopes that act as vaccine targets. This study describes current methodologies for the selection process, with Ebola virus as a model system. Hence great challenge in the field of ebola virus research is to design universal vaccine. A combination of publicly available bioinformatics algorithms and computational tools are used to screen and select antigen sequences as potential T-cell epitopes of supertypes Human Leukocyte Antigen (HLA) alleles. MUSCLE and MOTIF tools were used to find out most conserved peptide sequences of viral proteins. Immunoinformatics tools were used for prediction of immunogenic peptides of viral proteins in zaire strains of Ebola virus. Putative epitopes for viral proteins (VP) were predicted from conserved peptide sequences of VP. Three tools NetCTL 1.2, BIMAS and Syfpeithi were used to predict the Class I putative epitopes while three tools, ProPred, IEDB-SMM-align and NetMHCII 2.2 were used to predict the Class II putative epitopes. B cell epitopes were predicted by BCPREDS 1.0. Immunogenic peptides were identified and selected manually by putative epitopes predicted from online tools individually for both MHC classes. Finally sequences of predicted peptides for both MHC classes were looked for common region which was selected as common immunogenic peptide. The immunogenic peptides were found for viral proteins of Ebola virus: epitopes FLESGAVKY, SSLAKHGEY. These predicted peptides could be promising candidates to be used as target for vaccine design.Keywords: epitope, b cell, immunogenicity, ebola
Procedia PDF Downloads 3143897 Protein-Enrichment of Oilseed Meals by Triboelectrostatic Separation
Authors: Javier Perez-Vaquero, Katryn Junker, Volker Lammers, Petra Foerst
Abstract:
There is increasing importance to accelerate the transition to sustainable food systems by including environmentally friendly technologies. Our work focuses on protein enrichment and fractionation of agricultural side streams by dry triboelectrostatic separation technology. Materials are fed in particulate form into a system dispersed in a highly turbulent gas stream, whereby the high collision rate of particles against surfaces and other particles greatly enhances the electrostatic charge build-up over the particle surface. A subsequent step takes the charged particles to a delimited zone in the system where there is a highly uniform, intense electric field applied. Because the charge polarity acquired by a particle is influenced by its chemical composition, morphology, and structure, the protein-rich and fiber-rich particles of the starting material get opposite charge polarities, thus following different paths as they move through the region where the electric field is present. The output is two material fractions, which differ in their respective protein content. One is a fiber-rich, low-protein fraction, while the other is a high-protein, low-fiber composition. Prior to testing, materials undergo a milling process, and some samples are stored under controlled humidity conditions. In this way, the influence of both particle size and humidity content was established. We used two oilseed meals: lupine and rapeseed. In addition to a lab-scale separator to perform the experiments, the triboelectric separation process could be successfully scaled up to a mid-scale belt separator, increasing the mass feed from g/sec to kg/hour. The triboelectrostatic separation technology opens a huge potential for the exploitation of so far underutilized alternative protein sources. Agricultural side-streams from cereal and oil production, which are generated in high volumes by the industries, can further be valorized by this process.Keywords: bench-scale processing, dry separation, protein-enrichment, triboelectrostatic separation
Procedia PDF Downloads 1903896 A Microfluidic Biosensor for Detection of EGFR 19 Deletion Mutation Targeting Non-Small Cell Lung Cancer on Rolling Circle Amplification
Authors: Ji Su Kim, Bo Ram Choi, Ju Yeon Cho, Hyukjin Lee
Abstract:
Epidermal growth factor receptor (EGFR) 19 deletion mutation gene is over-expressed in carcinoma patient. EGFR 19 deletion mutation is known as typical biomarker of non-small cell lung cancer (NSCLC), which one section in the coding exon 19 of EGFR is deleted. Therefore, there have been many attempts over the years to detect EGFR 19 deletion mutation for replacing conventional diagnostic method such as PCR and tissue biopsy. We developed a simple and facile detection platform based on Rolling Circle Amplification (RCA), which provides highly amplified products in isothermal amplification of the ligated DNA template. Limit of detection (~50 nM) and a faster detection time (~30 min) could be achieved by introducing RCA.Keywords: EGFR19, cancer, diagnosis, rolling circle amplification (RCA), hydrogel
Procedia PDF Downloads 2553895 Effects of Bipolar Plate Coating Layer on Performance Degradation of High-Temperature Proton Exchange Membrane Fuel Cell
Authors: Chen-Yu Chen, Ping-Hsueh We, Wei-Mon Yan
Abstract:
Over the past few centuries, human requirements for energy have been met by burning fossil fuels. However, exploiting this resource has led to global warming and innumerable environmental issues. Thus, finding alternative solutions to the growing demands for energy has recently been driving the development of low-carbon and even zero-carbon energy sources. Wind power and solar energy are good options but they have the problem of unstable power output due to unpredictable weather conditions. To overcome this problem, a reliable and efficient energy storage sub-system is required in future distributed-power systems. Among all kinds of energy storage technologies, the fuel cell system with hydrogen storage is a promising option because it is suitable for large-scale and long-term energy storage. The high-temperature proton exchange membrane fuel cell (HT-PEMFC) with metallic bipolar plates is a promising fuel cell system because an HT-PEMFC can tolerate a higher CO concentration and the utilization of metallic bipolar plates can reduce the cost of the fuel cell stack. However, the operating life of metallic bipolar plates is a critical issue because of the corrosion phenomenon. As a result, in this work, we try to apply different coating layer on the metal surface and to investigate the protection performance of the coating layers. The tested bipolar plates include uncoated SS304 bipolar plates, titanium nitride (TiN) coated SS304 bipolar plates and chromium nitride (CrN) coated SS304 bipolar plates. The results show that the TiN coated SS304 bipolar plate has the lowest contact resistance and through-plane resistance and has the best cell performance and operating life among all tested bipolar plates. The long-term in-situ fuel cell tests show that the HT-PEMFC with TiN coated SS304 bipolar plates has the lowest performance decay rate. The second lowest is CrN coated SS304 bipolar plate. The uncoated SS304 bipolar plate has the worst performance decay rate. The performance decay rates with TiN coated SS304, CrN coated SS304 and uncoated SS304 bipolar plates are 5.324×10⁻³ % h⁻¹, 4.513×10⁻² % h⁻¹ and 7.870×10⁻² % h⁻¹, respectively. In addition, the EIS results indicate that the uncoated SS304 bipolar plate has the highest growth rate of ohmic resistance. However, the ohmic resistance with the TiN coated SS304 bipolar plates only increases slightly with time. The growth rate of ohmic resistances with TiN coated SS304, CrN coated SS304 and SS304 bipolar plates are 2.85×10⁻³ h⁻¹, 3.56×10⁻³ h⁻¹, and 4.33×10⁻³ h⁻¹, respectively. On the other hand, the charge transfer resistances with these three bipolar plates all increase with time, but the growth rates are all similar. In addition, the effective catalyst surface areas with all bipolar plates do not change significantly with time. Thus, it is inferred that the major reason for the performance degradation is the elevated ohmic resistance with time, which is associated with the corrosion and oxidation phenomena on the surface of the stainless steel bipolar plates.Keywords: coating layer, high-temperature proton exchange membrane fuel cell, metallic bipolar plate, performance degradation
Procedia PDF Downloads 2813894 Scale Effects on the Wake Airflow of a Heavy Truck
Authors: Aude Pérard Lecomte, Georges Fokoua, Amine Mehel, Anne Tanière
Abstract:
Air quality in urban areas is deteriorated by pollution, mainly due to the constant increase of the traffic of different types of ground vehicles. In particular, particulate matter pollution with important concentrations in urban areas can cause serious health issues. Characterizing and understanding particle dynamics is therefore essential to establish recommendations to improve air quality in urban areas. To analyze the effects of turbulence on particulate pollutants dispersion, the first step is to focus on the single-phase flow structure and turbulence characteristics in the wake of a heavy truck model. To achieve this, Computational Fluid Dynamics (CFD) simulations were conducted with the aim of modeling the wake airflow of a full- and reduced-scale heavy truck. The Reynolds Average Navier-Stokes (RANS) approach with the Reynolds Stress Model (RSM)as the turbulence model closure was used. The simulations highlight the apparition of a large vortex coming from the under trailer. This vortex belongs to the recirculation region, located in the near-wake of the heavy truck. These vortical structures are expected to have a strong influence on particle dynamics that are emitted by the truck.Keywords: CDF, heavy truck, recirculation region, reduced scale
Procedia PDF Downloads 2183893 Autophagy Regulates Human Hepatocellular Carcinoma Tumorigenesis through Selective Degradation of Cyclin D1
Authors: Shan-Ying Wu, Sheng-Hui Lan, Xi-Zhang Lin, Ih-Jen Su, Ting-Fen Tsai, Chia-Jui Yen, Tsung-Hsueh Lu, Fu-Wen Liang, Huey-Jen Su, Chun-Li Su, Hsiao-Sheng Liu
Abstract:
In hepatocelluar carcinoma (HCC), dysregulated expression of cyclin D1 and impaired autophagy has been reported separately. However, the relationship between them has not been explored. In this study, we demonstrated that autophagy was inversely correlated with cyclin D1 expression in 147 paired HCC patient specimens. HCC specimen with highly expression of cyclin D1 shows correlation with poor overall survival rate. Furthermore, induction of autophagy by amiodarone (antiarrhythmic drug) in Hep 3B cells, cyclin D1 was recruited into autophagosomes demonstrated by immune-gold labeling of cyclin D1 after extraction of autophagosomes. We further demonstrated that autophagy suppresses Hep 3B cell proliferation, and further analysis revealed that cell cycle was arrested at G1 phase. The interaction between LC3 (maker of autophagy) and cyclin D1 was increased after autophagy induction. In addition, ubiquitinated-cyclin D1 was also increased after autophagy induction, which is selectively degraded by autophagosome through binding with SQSTM1/p62 (an adaptor protein). In vivo study showed that amiodarone induced autophagy suppresses liver tumor formation in xenograft mouse and orthotopic rat model through decreasing cyclin D1 expression and inhibition of cell proliferation. Altogether, we reveal a novel mechanism that ubiquitinated cyclin D1 degraded by autophagic pathway by p62 and amiodarone is a promising drug for targeting cyclin D1 in liver cancer therapy.Keywords: autophagy, cyclin D1, hepatocellular carcinoma, amiodarone
Procedia PDF Downloads 2953892 A Two-Stage Airport Ground Movement Speed Profile Design Methodology Using Particle Swarm Optimization
Authors: Zhang Tianci, Ding Meng, Zuo Hongfu, Zeng Lina, Sun Zejun
Abstract:
Automation of airport operations can greatly improve ground movement efficiency. In this paper, we study the speed profile design problem for advanced airport ground movement control and guidance. The problem is constrained by the surface four-dimensional trajectory generated in taxi planning. A decomposed approach of two stages is presented to solve this problem efficiently. In the first stage, speeds are allocated at control points which ensure smooth speed profiles can be found later. In the second stage, detailed speed profiles of each taxi interval are generated according to the allocated control point speeds with the objective of minimizing the overall fuel consumption. We present a swarm intelligence based algorithm for the first-stage problem and a discrete variable driven enumeration method for the second-stage problem since it only has a small set of discrete variables. Experimental results demonstrate the presented methodology performs well on real world speed profile design problems.Keywords: airport ground movement, fuel consumption, particle swarm optimization, smoothness, speed profile design
Procedia PDF Downloads 5823891 Studies on Organic and Inorganic Micro/Nano Particle Reinforced Epoxy Composites
Authors: Daniel Karthik, Vijay Baheti, Jiri Militky, Sundaramurthy Palanisamy
Abstract:
Fibre based nano particles are presently considered as one of the potential filler materials for the improvement of mechanical and physical properties of polymer composites. Due to high matrix-filler interfacial area there will be uniform and homogeneous dispersion of nanoparticles. In micro/nano filler reinforced composites, resin material is usually tailored by organic or inorganic nanoparticles to have improved matrix properties. The objective of this study was to compare the potential of reinforcement of different organic and inorganic micro/nano fillers in epoxy composites. Industrial and agricultural waste of fibres like Agave Americana, cornhusk, jute, basalt, carbon, glass and fly ash was utilized to prepare micro/nano particles. Micro/nano particles were obtained using high energy planetary ball milling process in dry condition. Milling time and ball size were kept constant throughout the ball milling process. Composites were fabricated by hand lay method. Particle loading was kept constant to 3% wt. for all composites. In present study, loading of fillers was selected as 3 wt. % for all composites. Dynamic mechanical properties of the nanocomposite films were performed in three-point bending mode with gauge length and sample width of 50 mm and 10 mm respectively. The samples were subjected to an oscillating frequency of 1 Hz, 5 Hz and 10 Hz and 100 % oscillating amplitude in the temperature ranges of 30°C to 150°C at the heating rate of 3°C/min. Damping was found to be higher with the jute composites. Amongst organic fillers lowest damping factor was observed with Agave Americana particles, this means that Agave americana fibre particles have betters interface adhesion with epoxy resin. Basalt, fly ash and glass particles have almost similar damping factors confirming better interface adhesion with epoxy.Keywords: ball milling, damping factor, matrix-filler interface, particle reinforcements
Procedia PDF Downloads 2643890 A Comparative Study on the Thermophysical and Lubricity Characteristics of Multiwall Carbon Nanotube/Oil and Nanoclay/Oil Nanofluids
Authors: H. Singh, H. Bhowmick
Abstract:
Now-a-days, particle based lubricants have been widely used to enhance the lubrication performance. Use of tailor made micro/nanofluids can reduce the friction losses and dissipate heat in a better way. Use of Carbon Nanotubes (CNTs) has gained interests because of its structure that can endure much better in a system mechanically or thermally in comparison to any other additive in oil. On the other hand, nanoclays have been characterized mechanically and tribologically for the use of clay/polymer composite, and they have been gaining huge interest. Hence it is interesting to be investigated the effect of nanoclays as additive in oil. Thermophysical characteristics of lubricant play a predominant role in defining the friction and wear characteristics of lubricated contacts. However, very limited studies have been carried out to correlate the thermophysical properties of nanolubricants with their lubricity characteristics. Besides, most of the lubricant formulations till dates are found to be optimized for steel/steel contacts. In the present study, Multiwall Carbon Nanotube (MWCNT) and nanoclay are used as particle additives in mineral oil to develop nanofluids of various concentrations. The prepared lubricants are tested for their rheological, thermal and lubricity characteristics under aluminium-steel contacts. From the thermophysical investigation, it is observed that nanoclay particles significantly improve the viscosity of lubricant with an insignificant improvement in thermal conductivity. On the other hand, MWCNT particles moderately increase the viscosity but significantly increase the thermal conductivity of the base oil. Frictional responses of the nanofluids are characterized using a Pin-on-Disc tribometer which reveal some interesting facts. The findings from this study will greatly aid in formulating the particle based lubricants for cutting fluid in metal forming industries as well as fully developed nanolubricants for aluminium and Aluminium Metal Matrix Composite (AMMC) tribocontact for the use in the automotive and their allied industries.Keywords: MWCNT, Multiwall Carbon Nanotube, nanoclay, nanolubricant, rheology, thermal conductivity
Procedia PDF Downloads 1403889 Apoptosis Activity of Persea declinata (Bl.) Kosterm Bark Methanolic Crude Extract
Authors: P. Narrima, C. Y. Looi, M. A. Mohd, H. M. Ali
Abstract:
Persea declinata (Bl.) Kosterm is a member of the Lauraceae family, widely distributed in Southeast Asia. It is from the same genus with avocado (Persea americana Mill), which is widely consumed as food and for medicinal purposes. In the present study, we examined the anticancer properties of Persea declinata (Bl.) Kosterm bark methanolic crude extract (PDM). PDM exhibited a potent antiproliferative effect in MCF-7 human breast cancer cells, with an IC50 value of 16.68 µg/mL after 48h of treatment. We observed that PDM caused cell cycle arrest and subsequent apoptosis in MCF-7 cells, as exhibited by increased population at G0/G1 phase, higher lactate dehydrogenase (LDH) release, and DNA fragmentation. Mechanistic studies showed that PDM caused significant elevation in ROS production, leading to perturbation of mitochondrial membrane potential, cell permeability, and activation of caspases-3/7. On the other hand, real-time PCR and Western blot analysis showed that PDM treatment increased the expression of the proapoptotic molecule, Bax, but decreased the expression of prosurvival proteins, Bcl-2 and Bcl-xL, in a dose-dependent manner. These findings imply that PDM could inhibit proliferation in MCF-7 cells via cell cycle arrest and apoptosis induction, indicating its potential as a therapeutic agent worthy of further development.Keywords: antiproliferative, apoptosis, MCF-7 human breast cancer, Persea declinata
Procedia PDF Downloads 2433888 Effect of Helical Flow on Separation Delay in the Aortic Arch for Different Mechanical Heart Valve Prostheses by Time-Resolved Particle Image Velocimetry
Authors: Qianhui Li, Christoph H. Bruecker
Abstract:
Atherosclerotic plaques are typically found where flow separation and variations of shear stress occur. Although helical flow patterns and flow separations have been recorded in the aorta, their relation has not been clearly clarified and especially in the condition of artificial heart valve prostheses. Therefore, an experimental study is performed to investigate the hemodynamic performance of different mechanical heart valves (MHVs), i.e. the SJM Regent bileaflet mechanical heart valve (BMHV) and the Lapeyre-Triflo FURTIVA trileaflet mechanical heart valve (TMHV), in a transparent model of the human aorta under a physiological pulsatile right-hand helical flow condition. A typical systolic flow profile is applied in the pulse-duplicator to generate a physiological pulsatile flow which thereafter flows past an axial turbine blade structure to imitate the right-hand helical flow induced in the left ventricle. High-speed particle image velocimetry (PIV) measurements are used to map the flow evolution. A circular open orifice nozzle inserted in the valve plane as the reference configuration initially replaces the valve under investigation to understand the hemodynamic effects of the entered helical flow structure on the flow evolution in the aortic arch. Flow field analysis of the open orifice nozzle configuration illuminates the helical flow effectively delays the flow separation at the inner radius wall of the aortic arch. The comparison of the flow evolution for different MHVs shows that the BMHV works like a flow straightener which re-configures the helical flow pattern into three parallel jets (two side-orifice jets and the central orifice jet) while the TMHV preserves the helical flow structure and therefore prevent the flow separation at the inner radius wall of the aortic arch. Therefore the TMHV is of better hemodynamic performance and reduces the pressure loss.Keywords: flow separation, helical aortic flow, mechanical heart valve, particle image velocimetry
Procedia PDF Downloads 1743887 Investigation of Supercapacitor Properties of Nanocomposites Obtained from Acid and Base-functionalized Multi-walled Carbon Nanotube (MWCNT) and Polypyrrole (PPy)
Authors: Feridun Demir, Pelin Okdem
Abstract:
Polymers are versatile materials with many unique properties, such as low density, reasonable strength, flexibility, and easy processability. However, the mechanical properties of these materials are insufficient for many engineering applications. Therefore, there is a continuous search for new polymeric materials with improved properties. Polymeric nanocomposites are an advanced class of composite materials that have attracted great attention in both academic and industrial fields. Since nano-reinforcement materials are very small in size, they provide ultra-large interfacial area per volume between the nano-element and the polymer matrix. This allows the nano-reinforcement composites to exhibit enhanced toughness without compromising hardness or optical clarity. PPy and MWCNT/PPy nanocomposites were synthesized by the chemical oxidative polymerization method and the supercapacitor properties of the obtained nanocomposites were investigated. In addition, pure MWCNT was functionalized with acid (H₂SO₄/H₂O₂) and base (NH₄OH/H₂O₂) solutions at a ratio of 3:1 and a-MWCNT/d-PPy, and b-MWCNT/d-PPy nanocomposites were obtained. The homogeneous distribution of MWCNTs in the polypyrrole matrix and shell-core type morphological structures of the nanocomposites was observed with SEM images. It was observed with SEM, FTIR and XRD analyses that the functional groups formed by the functionalization of MWCNTs caused the MWCNTs to come together and partially agglomerate. It was found that the conductivity of the nanocomposites consisting of MWCNT and d-PPy was higher than that of pure d-PPy. CV, GCD and EIS results show that the use of a-MWCNT and b-MWCNTs in nanocomposites with low particle content positively affects the supercapacitor properties of the materials but negatively at high particle content. It was revealed that the functional MWCNT particles combined in nanocomposites with high particle content cause a decrease in the conductivity and distribution of ions in the electrodes and, thus, a decrease in their energy storage capacity.Keywords: polypyrrole, multi-walled carbon nanotube (MWCNT), conducting polymer, chemical oxidative polymerization, nanocomposite, supercapacitor
Procedia PDF Downloads 213886 In vitro Study of Laser Diode Radiation Effect on the Photo-Damage of MCF-7 and MCF-10A Cell Clusters
Authors: A. Dashti, M. Eskandari, L. Farahmand, P. Parvin, A. Jafargholi
Abstract:
Breast Cancer is one of the most considerable diseases in the United States and other countries and is the second leading cause of death in women. Common breast cancer treatments would lead to adverse side effects such as loss of hair, nausea, and weakness. These complications arise because these cancer treatments damage some healthy cells while eliminating the cancer cells. In an effort to address these complications, laser radiation was utilized and tested as a targeted cancer treatment for breast cancer. In this regard, tissue engineering approaches are being employed by using an electrospun scaffold in order to facilitate the growth of breast cancer cells. Polycaprolacton (PCL) was used as a material for scaffold fabricating because of its biocompatibility, biodegradability, and supporting cell growth. The specific breast cancer cells have the ability to create a three-dimensional cell cluster due to the spontaneous accumulation of cells in the porosity of the scaffold under some specific conditions. Therefore, we are looking for a higher density of porosity and larger pore size. Fibers showed uniform diameter distribution and final scaffold had optimum characteristics with approximately 40% porosity. The images were taken by SEM and the density and the size of the porosity were determined with the Image. After scaffold preparation, it has cross-linked by glutaraldehyde. Then, it has been washed with glycine and phosphate buffer saline (PBS), in order to neutralize the residual glutaraldehyde. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromidefor (MTT) results have represented approximately 91.13% viability of the scaffolds for cancer cells. In order to create a cluster, Michigan Cancer Foundation-7 (MCF-7, breast cancer cell line) and Michigan Cancer Foundation-10A (MCF-10A, human mammary epithelial cell line) cells were cultured on the scaffold in 24 well plate for five days. Then, we have exposed the cluster to the laser diode 808 nm radiation to investigate the effect of laser on the tumor with different power and time. Under the same conditions, cancer cells lost their viability more than the healthy ones. In conclusion, laser therapy is a viable method to destroy the target cells and has a minimum effect on the healthy tissues and cells and it can improve the other method of cancer treatments limitations.Keywords: breast cancer, electrospun scaffold, polycaprolacton, laser diode, cancer treatment
Procedia PDF Downloads 1433885 Combining in vitro Protein Expression with AlphaLISA Technology to Study Protein-Protein Interaction
Authors: Shayli Varasteh Moradi, Wayne A. Johnston, Dejan Gagoski, Kirill Alexandrov
Abstract:
The demand for a rapid and more efficient technique to identify protein-protein interaction particularly in the areas of therapeutics and diagnostics development is growing. The method described here is a rapid in vitro protein-protein interaction analysis approach based on AlphaLISA technology combined with Leishmania tarentolae cell-free protein production (LTE) system. Cell-free protein synthesis allows the rapid production of recombinant proteins in a multiplexed format. Among available in vitro expression systems, LTE offers several advantages over other eukaryotic cell-free systems. It is based on a fast growing fermentable organism that is inexpensive in cultivation and lysate production. High integrity of proteins produced in this system and the ability to co-express multiple proteins makes it a desirable method for screening protein interactions. Following the translation of protein pairs in LTE system, the physical interaction between proteins of interests is analysed by AlphaLISA assay. The assay is performed using unpurified in vitro translation reaction and therefore can be readily multiplexed. This approach can be used in various research applications such as epitope mapping, antigen-antibody analysis and protein interaction network mapping. The intra-viral protein interaction network of Zika virus was studied using the developed technique. The viral proteins were co-expressed pair-wise in LTE and all possible interactions among viral proteins were tested using AlphaLISA. The assay resulted to the identification of 54 intra-viral protein-protein interactions from which 19 binary interactions were found to be novel. The presented technique provides a powerful tool for rapid analysis of protein-protein interaction with high sensitivity and throughput.Keywords: AlphaLISA technology, cell-free protein expression, epitope mapping, Leishmania tarentolae, protein-protein interaction
Procedia PDF Downloads 2373884 DOG1 Expression Is in Common Human Tumors: A Tissue Microarray Study on More than 15,000 Tissue Samples
Authors: Kristina Jansen, Maximilian Lennartz, Patrick Lebok, Guido Sauter, Ronald Simon, David Dum, Stefan Steurer
Abstract:
DOG1 (Discovered on GIST1) is a voltage-gated calcium-activated chloride and bicarbonate channel that is highly expressed in interstitial cells of Cajal and in gastrointestinal stromal tumors (GIST) derived from Cajal cells. To systematically determine in what tumor entities and normal tissue types DOG1 may be further expressed, a tissue microarray (TMA) containing 15,965 samples from 121 different tumor types and subtypes as well as 608 samples of 76 different normal tissue types were analyzed by immunohistochemistry. DOG1 immunostaining was found in 67 tumor types, including GIST (95.7%), esophageal squamous cell carcinoma (31.9%), pancreatic ductal adenocarcinoma (33.6%), adenocarcinoma of the Papilla Vateri (20%), squamous cell carcinoma of the vulva (15.8%) and the oral cavity (15.3%), mucinous ovarian cancer (15.3%), esophageal adenocarcinoma (12.5%), endometrioid endometrial cancer (12.1%), neuroendocrine carcinoma of the colon (11.1%) and diffuse gastric adenocarcinoma (11%). Low level-DOG1 immunostaining was seen in 17 additional tumor entities. DOG1 expression was unrelated to histopathological parameters of tumor aggressiveness and/or patient prognosis in cancers of the breast (n=1,002), urinary bladder (975), ovary (469), endometrium (173), stomach (233), and thyroid gland (512). High DOG1 expression was linked to estrogen receptor expression in breast cancer (p<0.0001) and the absence of HPV infection in squamous cell carcinomas (p=0.0008). In conclusion, our data identify several tumor entities that can show DOG1 expression levels at similar levels as in GIST. Although DOG1 is tightly linked to a diagnosis of GIST in spindle cell tumors, the differential diagnosis is much broader in DOG1 positive epithelioid neoplasms.Keywords: biomarker, DOG1, immunohistochemistry, tissue microarray
Procedia PDF Downloads 2163883 Adaptive Cooperative Scheme Considering the User Location
Authors: Bit-Na Kwon, Hyun-Jee Yang, Dong-Hyun Ha, Hyoung-Kyu Song
Abstract:
In this paper, an adaptive cooperative scheme in the cell edge is proposed. The proposed scheme considers the location of a user and applies the suitable cooperative scheme. In cellular systems, the performance of communication is degraded if the user is located in the cell edge. In conventional scheme, two base stations are used in order to obtain diversity gain. However, the performance of communication is not sufficiently improved since the distance between each base station and a user is still distant. Therefore, we propose a scheme that the relays are used and the cooperative scheme is adaptively applied according to the user location. Through simulation results, it is confirmed that the proposed scheme has better performance than the conventional scheme.Keywords: adaptive transmission, cooperative communication, diversity gain, OFDM
Procedia PDF Downloads 5723882 The Creation of a Yeast Model for 5-oxoproline Accumulation
Authors: Pratiksha Dubey, Praveen Singh, Shantanu Sen Gupta, Anand K. Bachhawat
Abstract:
5-oxoproline (pyroglutamic acid) is a cyclic lactam of glutamic acid. In the cell, it can be produced by several different pathways and is metabolized into glutamate with the help of the 5-oxoprolinase enzyme (OPLAH or OXP1). The inhibition of 5-oxoprolinase enzyme in mammals was found to result in heart failure and is thought to be a consequence of oxidative stress [1]. To analyze the consequences of 5-oxoproline accumulation more clearly, we are generating models for 5-oxoproline accumulation in yeast. The 5-oxoproline accumulation model in yeast is being developed by two different strategies. The first one is by overexpression of the mouse -glutamylcyclotransferase enzyme. It degrades -glu-met dipeptide into 5-oxoproline and methionine taken by the cell from the medium. The second strategy is by providing high concentration of 5-oxoproline externally to the yeast cells. The intracellular 5-oxoproline levels in both models are being evaluated. In addition, the metabolic and cellular consequences are being investigated.Keywords: 5-oxoproline, pyroglutamic acid, yeast, genetics
Procedia PDF Downloads 873881 The Effect of Size and Tumor Depth on Histological Clearance Margins of Basal Cell Carcinomas
Authors: Martin Van, Mohammed Javed, Sarah Hemington-Gorse
Abstract:
Aim: Our aim was to determine the effect of size and tumor depth of basal cell carcinomas (BCCs) on surgical margin clearance. Methods: A retrospective study was conducted at the Welsh Centre for Burns and Plastic Surgery (WCBPS), Morriston Hospital between 1 Jan 2016 – 31 July 2016. Only patients with confirmed BCC on histopathological analysis were included. Patient data including anatomical region treated, lesion size, histopathological clearance margins and histological sub-types were recorded. An independent T-test was performed determine statistical significance. Results: A total of 228 BCCs were excised in 160 patients. Eleven lesions (4.8%) were incompletely excised. The nose area had the highest rate of incomplete excision. The mean diameter of incompletely excised lesions was 11.4mm vs 11.5mm in completely excised lesions (p=0.959) and the mean histological depth of incompletely excised lesions was 4.1mm vs. 2.5mm for completely excised BCCs (p < 0.05). Conclusions: BCC tumor depth of > 4.1 mm was associated with high rate of incomplete margin clearance. Hence, in prospective patients, a BCC tumor depth (>4 mm) on tissue biopsy should alert the surgeon of potentially higher risk of incomplete excision of lesion.Keywords: basal cell carcinoma, excision margins, plastic surgery, treatment
Procedia PDF Downloads 2383880 Cytotoxic Activity of Parkia javanica Merr. and Parkia speciosa Hassk. against Human Cancer Cell Lines
Authors: Srisopa Ruangnoo, Arunporn Itharat
Abstract:
The ethanolic and aqueous extracts of Parkia javanica Merr. germinating seeds and Parkia speciosa Hassk. seeds were evaluated for cytotoxic activity against three different types of human cancer cell lines including colon cancer (LS174T), breast cancer (MCF-7) and prostate cancer (PC3) using sulforhodamine B (SRB) assay. The fresh plant parts were divided into 2 parts. The first part was extracted by maceration with 95% ethanol for 3 days and then filtered, and the filtrates were evaporated by rotary evaporator. The other part was squeezed and filtered. Then the filtrates were dried by freeze dryer. The screening found that the aqueous extract of P. javanica Merr. germinating seeds exhibited more than 70% inhibition (at concentration 50 µg/ml) against all types of human cancer cells. The aqueous extract of P. javanica Merr. germinating seeds showed the highest cytotoxic activity against MCF-7 with the IC50 value as 5.63 µg/ml. The aqueous extract of P. javanica Merr. germinating seeds also showed high cytotoxic activity against PC3 and LS174T with the IC50 values as 10.79 and 11.40 µg/ml, respectively. In conclusion, P. javanica Merr. germinating seed is a natural source of anticancer activity and further research to isolate active compounds from this plant should be undertaken.Keywords: cytotoxic activity, Parkia javanica Merr., Parkia speciosa Hassk., human cancer cell lines
Procedia PDF Downloads 4083879 Comparison of Physical and Chemical Effects on Senescent Cells
Authors: Svetlana Guryeva, Inna Kornienko, Andrey Usanov, Dmitry Usanov, Elena Petersen
Abstract:
Every day cells in our organism are exposed to various factors: chemical agents, reactive oxygen species, ionizing radiation, and others. These factors can cause damage to DNA, cellular membrane, intracellular compartments, and proteins. The fate of cells depends on the exposure intensity and duration. The prolonged and intense exposure causes the irreversible damage accumulation, which triggers the permanent cell cycle arrest (cellular senescence) or cell death programs. In the case of low dose of impacts, it can lead to cell renovation and to cell functional state improvement. Therefore, it is a pivotal question to investigate the factors and doses that result in described positive effects. In order to estimate the influence of different agents, the proliferation index and levels of cell death markers (annexin V/propidium iodide), senescence-associated β-galactosidase, and lipofuscin were measured. The experiments were conducted on primary human fibroblasts of the 8th passage. According to the levels of mentioned markers, these cells were defined as senescent cells. The effect of low-frequency magnetic field was investigated. Different modes of magnetic field exposure were tested. The physical agents were compared with chemical agents: metformin (10 mM) and taurine (0.8 mM and 1.6 mM). Cells were incubating with chemicals for 5 days. The highest decrease in the level of senescence-associated β-galactosidase (21%) and lipofuscin (17%) was observed in the primary senescent fibroblasts after 5 days after double treatments with 48 h intervals with low-frequency magnetic field. There were no significant changes in the proliferation index after magnetic field application. The cytotoxic effect of magnetic field was not observed. The chemical agent taurine (1.6 mM) decreased the level of senescence-associated β-galactosidase (23%) and lipofuscin (22%). Metformin improved the activity of senescence-associated β-galactosidase on 15% and the level of lipofuscin on 19% in this experiment. According to these results, the effect of double treatment with 48 h interval with low-frequency magnetic field and the effect of taurine (1.6 mM) were comparable to the effect of metformin, for which anti-aging properties are proved. In conclusion, this study can become the first step towards creation of the standardized system for the investigation of different effects on senescent cells.Keywords: biomarkers, magnetic field, metformin, primary fibroblasts, senescence, taurine
Procedia PDF Downloads 2803878 Designing of Multi-Epitope Peptide Vaccines for Fasciolosis (Fasciola gigantica) using Immune Epitope and Analysis Resource (IEDB) Server
Authors: Supanan Chansap, Werachon Cheukamud, Pornanan Kueakhai, Narin Changklungmoa
Abstract:
Fasciola species (Fasciola spp.) is caused fasciolosis in ruminants such as cattle, sheep, and buffalo. Fasciola gigantica (F.gigantica) commonly infects tropical regions. Fasciola hepatica (F.hepatica) in temperate regions. Liver fluke infection affects livestock economically, for example, reduced milk and meat production, weight loss, sterile animals. Currently, Triclabendazole is used to treat liver flukes. However, liver flukes have also been found to be resistant to drugs in countries. Therefore, vaccination is an attractive alternative to prevent liver fluke infection. Peptide vaccines are new vaccine technologies that mimic epitope antigens that trigger an immune response. An interesting antigen used in vaccine production is catepsin L, a family of proteins that play an important role in the life of the parasite in the host. This study aims to identify immunogenic regions of protein and construct a multi-epidetope vaccine using an immunoinformatic tool. Fasciola gigantica Cathepsin L1 (FgCatL1), Fasciola gigantica Cathepsin L1G (FgCatL1G), and Fasciola gigantica Cathepsin L1H (FgCatL1H) were predicted B-cell and Helper T lymphocytes (HTL) by Immune Epitope and Analysis Resource (IEDB) servers. Both B-cell and HTL epitopes aligned with cathepsin L of the host and Fasciola hepatica (F. hepatica). Epitope groups were selected from non-conserved regions and overlapping sequences with F. hepatica. All overlapping epitopes were linked with the GPGPG and KK linker. GPGPG linker was linked between B-cell epitope. KK linker was linked between HTL epitope and B-cell and HTL epitope. The antigenic scores of multi-epitope peptide vaccine was 0.7824. multi-epitope peptide vaccine was non-allergen, non-toxic, and good soluble. Multi-epitope peptide vaccine was predicted tertiary structure and refinement model by I-Tasser and GalaxyRefine server, respectively. The result of refine structure model was good quality that was generated by Ramachandran plot analysis. Discontinuous and linear B-cell epitopes were predicted by ElliPro server. Multi-epitope peptide vaccine model was two and seven of discontinuous and linear B-cell epitopes, respectively. Furthermore, multi-epitope peptide vaccine was docked with Toll-like receptor 2 (TLR-2). The lowest energy ranged from -901.3 kJ/mol. In summary, multi-epitope peptide vaccine was antigenicity and probably immune response. Therefore, multi-epitope peptide vaccine could be used to prevent F. gigantica infections in the future.Keywords: fasciola gigantica, Immunoinformatic tools, multi-epitope, Vaccine
Procedia PDF Downloads 793877 Excellent Outcome with Early Diagnosis in an Infant with Wiskott-Aldrich Syndrome in a Tertiary Hospital in Oman
Authors: Surekha Tony, Roshan Mevada
Abstract:
Wiskott-Aldrich syndrome (WAS) is a primary immunodeficiency disease resulting in recurrent infections, eczema, and microthrombocytopenia. In its classical form, significant combined immune deficiency, autoimmune complications, and risk of hematological malignancy necessitate early correction, preferably before 2 years of age, with hematopoietic stem cell transplant (HSCT) or gene therapy. Clinical features and severity are varied, making the diagnosis difficult in milder cases. We report an Omani boy diagnosed in early infancy with WAS based on clinical presentation and confirmed by genetic diagnosis with cure by HSCT from an HLA-identical sibling donor.Keywords: genetic diagnosis, hematopoietic stem cell transplant, infant, Wiskott-Aldrich syndrome
Procedia PDF Downloads 183876 Investigation of Astrocyte Physiology on Stiffness-Controlled Cellulose Acetate Nanofiber as a Tissue Scaffold
Authors: Sun Il Yu, Jung Hyun Joo, Hwa Sung Shin
Abstract:
Astrocytes are known as dominant cells in CNS and play a role as a supporter of CNS activity and regeneration. Recently, three-dimensional culture of astrocytes were actively applied to understand in vivo astrocyte works. Electrospun nanofibers are attractive for 3D cell culture system because they have a high surface to volume ratio and porous structure, and have already been used for 3D astrocyte cultures. In this research, the stiffness of cellulose acetate (CA) nanofiber was controlled by heat treatment. As stiffness increased, astrocyte cell viability and adhesion increased. Reactivity of astrocyte was also upregulated in stiffer CA nanofiber in terms of GFAP, an intermediate filament protein. Finally, we demonstrated that stiffness-controllable CA is attractive for astrocyte tissue engineering.Keywords: astrocyte, cellulose acetate, nanofiber, tissue scaffold
Procedia PDF Downloads 3553875 Burn/Traumatic Scar Maturation Using Autologous Fat Grafts + SVF
Authors: Ashok K. Gupta
Abstract:
Over the past few decades, since the bio-engineering revolution, autologous cell therapy (ACT) has become a rapidly evolving field. Currently, this form of therapy has broad applications in modern medicine and plastic surgery, ranging from the treatment/improvement of wound healing to life-saving operations. A study was conducted on 50 patients having to disfigure, and deform post burn scars and was treated by injection of extracted, refined adipose tissue grafts with their unique stem cell properties. To compare the outcome, a control of 20 such patients was treated with conventional skin or soft-tissue flaps or skin grafting, and a control of 10 was treated with more advanced microsurgical techniques such as Pre-fabricated flaps/pre laminated flaps / free flaps. Assessment of fat volume and survival post- follow up period was done by radiological aid, using MRI and clinically (Survival of the autograft and objective parameters for scar elasticity were evaluated skin elasticity parameters 3 to 9 months postoperatively). Recently, an enzyme that is involved in collagen crosslinking in fibrotic tissue, lysyl hydroxylase (LH2), was identified. This enzyme is normally active in bone and cartilage but hardly in the skin. It has been found that this enzyme is highly expressed in scar tissue and subcutaneous fat; this is in contrast to the dermis, where the enzyme is hardly expressed. Adipose tissue-derived stem cell injections are an effective method in the treatment of various extensive post-burn scar deformities that makes it possible to re-create the lost sub-dermal tissue for improvement in the function of involved joint movements.Keywords: adipose tissue-derived stem cell injections, treatment of various extensive post-burn scar deformities, re-create the lost sub-dermal tissue, improvement in function of involved joint movements
Procedia PDF Downloads 673874 Mitigation of Size Effects in Woven Fabric Composites Using Finite Element Analysis Approach
Authors: Azeez Shaik, Yagnik Kalariya, Amit Salvi
Abstract:
High-performance requirements and emission norms were forcing the automobile industry to opt for lightweight materials which improve the fuel efficiency and absorb energy during crash applications. In such scenario, the woven fabric composites are providing better energy absorption compared to metals. Woven fabric composites have a repetitive unit cell (RUC) and the mechanical properties of these materials are highly dependent on RUC. This work investigates the importance of detailed modelling of the RUC, the size effects associated and the mitigation techniques to avoid them using Finite element analysis approach.Keywords: repetitive unit cell, representative volume element, size effects, cohesive zone, finite element analysis
Procedia PDF Downloads 2553873 In Vitro Intestine Tissue Model to Study the Impact of Plastic Particles
Authors: Ashleigh Williams
Abstract:
Micro- and nanoplastics’ (MNLPs) omnipresence and ecological accumulation is evident when surveying recent environmental impact studies. For example, in 2014 it was estimated that at least 52.3 trillion plastic microparticles are floating at sea, and scientists have even found plastics present remote Arctic ice and snow (5,6). Plastics have even found their way into precipitation, with more than 1000 tons of microplastic rain precipitating onto the Western United States in 2020. Even more recent studies evaluating the chemical safety of reusable plastic bottles found that hundreds of chemicals leached into the control liquid in the bottle (ddH2O, ph = 7) during a 24-hour time period. A consequence of the increased abundance in plastic waste in the air, land, and water every year is the bioaccumulation of MNLPs in ecosystems and trophic niches of the animal food chain, which could potentially cause increased direct and indirect exposure of humans to MNLPs via inhalation, ingestion, and dermal contact. Though the detrimental, toxic effects of MNLPs have been established in marine biota, much less is known about the potentially hazardous health effects of chronic MNLP ingestion in humans. Recent data indicate that long-term exposure to MNLPs could cause possible inflammatory and dysbiotic effects. However, toxicity seems to be largely dose-, as well as size-dependent. In addition, the transcytotic uptake of MNLPs through the intestinal epithelia in humans remain relatively unknown. To this point, the goal of the current study was to investigate the mechanisms of micro- and nanoplastic uptake and transcytosis of Polystyrene (PE) in human stem-cell derived, physiologically relevant in vitro intestinal model systems, and to compare the relative effect of particle size (30 nm, 100 nm, 500 nm and 1 µm), and concentration (0 µg/mL, 250 µg/mL, 500 µg/mL, 1000 µg/mL) on polystyrene MNLP uptake, transcytosis and intestinal epithelial model integrity. Observational and quantitative data obtained from confocal microscopy, immunostaining, transepithelial electrical resistance (TEER) measurements, cryosectioning, and ELISA cytokine assays of the proinflammatory cytokines Interleukin-6 and Interleukin-8 were used to evaluate the localization and transcytosis of polystyrene MNPs and its impact on epithelial integrity in human-derived intestinal in vitro model systems. The effect of Microfold (M) cell induction on polystyrene micro- and nanoparticle (MNP) uptake, transcytosis, and potential inflammation was also assessed and compared to samples grown under standard conditions. Microfold (M) cells, link the human intestinal system to the immune system and are the primary cells in the epithelium responsible for sampling and transporting foreign matter of interest from the lumen of the gut to underlying immune cells. Given the uptake capabilities of Microfold cells to interact both specifically and nonspecific to abiotic and biotic materials, it was expected that M- cell induced in vitro samples would have increased binding, localization, and potentially transcytosis of Polystyrene MNLPs across the epithelial barrier. Experimental results of this study would not only help in the evaluation of the plastic toxicity, but would allow for more detailed modeling of gut inflammation and the intestinal immune system.Keywords: nanoplastics, enteroids, intestinal barrier, tissue engineering, microfold (M) cells
Procedia PDF Downloads 853872 Epidemiology of Primary Bronchopulmonary Cancer in Tunisia
Authors: Melliti Rihab, Zaeid Sonia, Khechine Wiem, Daldoul Amira
Abstract:
Introduction: Lung cancer is the leading cause of cancer death. Its incidence is increasing, and its prognosis remains pejorative. We present the clinical, pathological, and therapeutic characteristics of bronchopulmonary cancer (BPC) in Tunisia. Methods: Retrospective study including patients followed in the oncology department of the University Hospital of Monastir between April 2014 and December 2021 suffering from lung cancer. Results: These are 117 patients, including 86.3% men and 13.7% women (sex ratio 6.3). The average age was 64 years ± 9 (37-83), with 95.7% being over 50 years old. Patients were smokers in 82% of cases. The clinical signs were dominated by chest pain (27.5%) and dyspnea in 21.1% of cases. In 6 patients, an episode of COVID-19 infection revealed the diagnosis. Half of the patients had a PS between 0 and 1. Small cell lung cancer was present in 18 patients (15.4%). The majority of non small cell lung cancer was of the adenocarcinoma type (68.7%). The diagnosis was late (stage IV) in 62.4% of cases. BPC was metastatic to bone (52%), contralateral lung (25.9%), and brain (27.3%). Patients were oligometastatic in 26% of cases. Surgery and radiotherapy were performed respectively in 14.5% and 23.1% of cases. Three-quarters of the patients had had nutrition (75.2%). The ROS1 mutation was present in 1 patient. PDL-1 expression was >40% in 2 patients. Survival was mean eight months ± 7.4. Conclusion: Lung cancer is diagnosed at a late stage in Tunisia. The lack of molecular study for non-small cell PBC and the lack of marketing authorization for tyrosine kinase inhibitors in Tunisia make the management incomplete.Keywords: SCLC, NCSLC, ROS1, PDL1
Procedia PDF Downloads 793871 Comparison of Entropy Coefficient and Internal Resistance of Two (Used and Fresh) Cylindrical Commercial Lithium-Ion Battery (NCR18650) with Different Capacities
Authors: Sara Kamalisiahroudi, Zhang Jianbo, Bin Wu, Jun Huang, Laisuo Su
Abstract:
The temperature rising within a battery cell depends on the level of heat generation, the thermal properties and the heat transfer around the cell. The rising of temperature is a serious problem of Lithium-Ion batteries and the internal resistance of battery is the main reason for this heating up, so the heat generation rate of the batteries is an important investigating factor in battery pack design. The delivered power of a battery is directly related to its capacity, decreases in the battery capacity means the growth of the Solid Electrolyte Interface (SEI) layer which is because of the deposits of lithium from the electrolyte to form SEI layer that increases the internal resistance of the battery. In this study two identical cylindrical Lithium-Ion (NCR18650)batteries from the same company with noticeable different in capacity (a fresh and a used battery) were compared for more focusing on their heat generation parameters (entropy coefficient and internal resistance) according to Brandi model, by utilizing potentiometric method for entropy coefficient and EIS method for internal resistance measurement. The results clarify the effect of capacity difference on cell electrical (R) and thermal (dU/dT) parameters. It can be very noticeable in battery pack design for its Safety.Keywords: heat generation, Solid Electrolyte Interface (SEI), potentiometric method, entropy coefficient
Procedia PDF Downloads 4733870 The Transcriptional Regulation of Human LRWD1 through DNA Methylation
Authors: Yen-Ni Teng, Hsing-Yi Chen, Hsien-An Pan, Yung-Ming Lin, Hany A. Omar, Jui-Hsiang Hung
Abstract:
Leucine-rich repeats and WD repeat domain containing 1 (LRWD1) is highly expressed in the testes of healthy males. On the other hand, LRWD1 is significantly down-regulated in the testicular tissues of patients with severe spermatogenic defects. In our study, the downregulation of LRWD1 expression by shRNA caused a significant reduction of cell growth and mitosis and a noteworthy increase in the cell microtubule atrophy rate. Here, we used EMBOSS CpG plot analysis to explore the promoter region of LRWD1 gene. We found that CpG islands are located between positions -253 to +5 nucleotides upstream from the LRWD1 transcription start site. Luciferase reporter assay revealed that the hypermethylation of the LRWD1 promoter reduced the transcription activity in cells. In addition, quantitative methylation-specific PCR and immunostaining showed that the methylation inhibitor, 5-Aza-2'-deoxycytidine, increased LRWD1 promoter activity, LRWD1 mRNA, protein expression and cell viability. Whereas, the methylation activator, S-adenosylmethionine, caused opposite effects. The overexpression of p53 and Nrf2 in NT2/D1 cells increased LRWD1 promoter activity while 5-fluorodeoxyuridine decreased it. In conclusion, this study highlights evidence that the methylation status of LRWD1 promoter is associated with LRWD1 expression. Since the expression level of LRWD1 plays an important role in spermatogenesis, the methylation status of LRWD1 may serve as a novel molecular diagnostic or therapeutic approach in male's infertility.Keywords: LRWD1, DNA methylation, p53, Nrf2
Procedia PDF Downloads 1483869 MAFB Expression in LPS-Induced Exosomes: Revealing the Connection to sepsis-trigerred Hepatic Injury
Authors: Gizaw Mamo Gebeyehu, Marianna Pap, Geza Makkai, Tibor Z. Janosi, Shima Rashidian, Tibor A. Rauch
Abstract:
Sepsis poses a significant global health threat, necessitating extensive exploration of indicators tied to its pathological mechanisms and multi-organ dysfunction. While murine studies have shed light on sepsis, the intricate cellular and molecular landscape in human sepsis remains enigmatic. Exploring the influence of activated monocyte-derived exosomes in sepsis sheds light on a promising pathway for understanding the intricate cellular and molecular mechanisms involved in this condition in humans. In sepsis, exosome-borne mRNA and miRNA orchestrate immune response gene expression in recipient cells. Yet, the specifics of exosome-mediated cell-to-cell communication, especially how mRNA cargoes modulate gene expression in recipient cells, remain poorly understood. This study aims to elucidate the precise molecular pathways through which exosomal mRNA cargo, particularly MAFB, contributes to the developing sepsis-induced molecular aberrations in liver tissues, employing rigorously defined cell culture conditions. THP-1 cells were treated with LPS to induce changes in exosomal RNA profiles. Exosomes were isolated and characterized using microscopy and mass spectrometry. RNA was extracted from exosomes and sequenced. The most abundant exosomal mRNAs were subjected to GO analysis for functional annotation analysis and KEGG database analysis to identify the involved enriched pathways. PCR (Polymerase Chain Reaction), RNA sequencing, and Western blotting were involved to analyze changes in gene expression, protein levels, and signaling pathways within the liver cells( HepG2) after exposure to exosomal MAFB. This study pinpoints exosomal MAFB as a potential key regulator linked to liver cell damage during sepsis, along with associated genes (miR155HG, H3F3A, and possibly JARD2) forming a crucial molecular pathway contributing to liver cell injury, Together, these elements indicate a vital molecular pathway that plays a significant role in the emergence of liver cell injury during sepsis.. These findings suggest the importance of further research on these components for potential therapeutic interventions in managing acute liver damage in sepsis.Keywords: sepsis, exososome, exosomal MAFB, LPS-induced THP-1 cells, RNA profiles, sepsis-triggered liver injury
Procedia PDF Downloads 64