Search results for: cell viability
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 4100

Search results for: cell viability

3110 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

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3109 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

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3108 Sustainable Cities: Viability of a Hybrid Aeroponic/Nutrient Film Technique System for Cultivation of Tomatoes

Authors: D. Dannehl, Z. Taylor, J. Suhl, L. Miranda, R., Ulrichs, C., Salazar, E. Fitz-Rodriguez, I. Lopez-Cruz, A. Rojano-Aguilar, G. Navas-Gomez, U. Schmidt

Abstract:

Growing environmental and sustainability concerns have driven continual modernization of horticultural practices, especially for urban farming. Controlled environment and soilless production methods are increasing in popularity because of their efficient resource use and intensive cropping capabilities. However, some popular substrates used for hydroponic cultivation, particularly rock wool, represent a large environmental burden in regard to their manufacture and disposal. Substrate-less hydroponic systems are effective in producing short cropping cycle plants such as lettuce or herbs, but less information is available for the production of plants with larger root-systems and longer cropping times. Here, we investigated the viability of a hybrid aeroponic/nutrient film technique (AP/NFT) system for the cultivation of greenhouse tomatoes (Solanum lycopersicum ‘Panovy’). The plants grown in the AP/NFT system had a more compact phenotype, accumulated more Na+ and less P and S than the rock wool grown counterparts. Due to forced irrigation interruptions, we propose that the differences observed were cofounded by the differing severity of water-stress for plants with and without substrate. They may also be caused by a higher root zone temperature predominant in plants exposed to AP/NFT. However, leaf area, stem diameter, and number of trusses did not differ significantly. The same was found for leaf pigments and plant photosynthetic efficiency. Overall, the AP/NFT system appears to be viable for the production of greenhouse tomato, enabling the environment to be relieved by way of lessening rock wool usage.

Keywords: closed aeroponic systems, fruit quality, nutrient dynamics, substrate waste reduction, urban farming systems, water savings

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3107 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

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3106 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

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3105 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

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3104 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

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3103 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

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3102 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

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3101 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

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3100 Insect Inducible Methanol Production in Plants for Insect Resistance

Authors: Gourav Jain, Sameer Dixit, Surjeet Kumar Arya, Praveen C. Verma

Abstract:

Plant cell wall plays a major role in defence mechanism against biotic and abiotic stress as it constitutes the physical barrier between the microenvironment and internal component of the cell. It is a complex structure composed of mostly carbohydrates among which cellulose and hemicelluloses are most abundant that is embedded in a matrix of pectins and proteins. Multiple enzymes have been reported which plays a vital role in cell wall modification, Pectin Methylesterase (PME) is one of them which catalyses the demethylesterification of homogalacturonans component of pectin which releases acidic pectin and methanol. As emitted methanol is toxic to the insect pest, we use PME gene for the better methanol production. In the current study we showed overexpression of PME gene isolated from Withania somnifera under the insect inducible promoter causes enhancement of methanol production at the time of insect feeds to plants, and that provides better insect resistance property. We found that the 85-90% mortality causes by transgenic tobacco in both chewing (Spodoptera litura larvae and Helicoverpa armigera) and sap-sucking (Aphid, mealybug, and whitefly) pest. The methanol content and emission level were also enhanced by 10-15 folds at different inducible time point interval (15min, 30min, 45min, 60min) which would be analysed by Purpald/Alcohol Oxidase method.

Keywords: methanol, Pectin methylesterase, inducible promoters, Purpald/Alcohol oxidase

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3099 Impact of Glaucoma Surgery on Corneal Endothelium

Authors: Majid Moshirfar, Kyle Margulies, Yasmyne C. Ronquillo, Phillip Hoopes

Abstract:

A total of 66 articles were reviewed to compare glaucoma and its associated surgeries' effect on central corneal endothelium cell density (CECD). The paper reports the average reported central CECD loss at 3-, 6-, 12-, 24-, 36-, 48-, and 60-month post-operation for each glaucoma surgery. ALT, MLT, SLT, CS AGV, VC BGI, Hydrus + phaco, XEN gel + phaco, PRESERFLO, Dual iStent, or Trabectome had no significant impact on postoperative CECD compared to either preoperative CECD or control group CECD. The highest CECD loss was found to be EXPRESS-phaco, AC AGV, CS BGI, CS BGI, AC BGI, and AC BGI at the 3-, 6-, 12-, 24-, 36-, 48-, and 60-month follow-ups, respectively. AC AGV, Trab + MMC, Trab, AC BGI, Trab + MMC, Cypass, and Cypass showed the smallest reduction of CECD at the 3-, 6-, 12-, 24-, 36-, 48-, and 60-month follow-ups.

Keywords: glaucoma, corneal endothelium, cell density, surgery outcome

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3098 Voltage Polarity in Electrospinning: Way to Control Surface Properties of Polymer Fibers

Authors: Urszula Stachewicz

Abstract:

Surface properties of materials are the key parameter in many applications, especially in the biomedical field, to control cell-material interactions. In our work, we want to achieve the controllability of surface properties of polymer fibers via a single-step electrospinning process by alternating voltage polarities. Voltage polarity defines the charge accumulated on the surface of the liquid jet and the surface of the fibers. Positive polarity attracts negatively charged groups to fibers’ surface, whereas negative polarity moves the negatively charged functional groups away from the surface. This way, we can control the surface chemistry, wettability, and additionally surface potential of electrospun fibers. Within our research, we characterized surface chemistry using X-ray photoelectron microscopy (XPS) and surface potential with Kelvin probe force microscopy (KPFM) on electrospun fibers of commonly used polymers such as PCL, PVDF, and PMMA, often used as biomaterials. We proved the significant effect of fibers' surface potential on cell integration with the scaffolds and further cells development for the regeneration processes based on the osteoblast and fibroblast culture studies. Acknowledgments: The study was conducted within ‘Nanofiber-based sponges for atopic skin treatment’ project, which is carried out within the First TEAM programme of the Foundation for Polish Science co-financed by the European Union under the European Regional Development Fund, project no POIR.04.04.00-00- 4571/18-00.

Keywords: cell attachment, fibers, fibroblasts, osteoblast, proliferation, surface potential

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3097 High Productivity Fed-Batch Process for Biosurfactant Production for Enhanced Oil Recovery Applications

Authors: G. A. Amin, A. D. Al-Talhi

Abstract:

The bacterium B. subtilis produced surfactin in conventional batch culture as a growth associated product and a growth rate (0.4 h-1). A fed-batch process was developed and the fermentative substrate and other nutrients were fed on hourly basis and according to the growth rate of the bacterium. Conversion of different quantities of Maldex-15 into surfactin was investigated in five different fermentation runs. In all runs, most of Maldex-15 was consumed and converted into surfactin and cell biomass with appreciable efficiencies. The best results were obtained with fermentation run supplied with 200 g Maldex-15. Up to 35.4 g.l-1 of surfactin and cell biomass of 30.2 g.l-1 were achieved in 12 hrs. Also, markedly substrate yield of 0.269 g/g and volumetric reactor productivity of 2.61 g.1-1.h-1 were obtained confirming the establishment of a cost effective commercial surfactin production.

Keywords: Bacillus subtilis, biosurfactant, exponentially fed-batch fermentation, surfactin

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3096 Electro-Winning of Dilute Solution of Copper Metal from Sepon Mine, Lao PDR

Authors: S. Vasailor, C. Rattanakawin

Abstract:

Electro-winning of copper metal from dilute sulfate solution (13.7 g/L) was performed in a lab electrolytic cell with stainless-steel cathode and lead-alloy anode. The effects of various parameters including cell voltage, electro-winning temperature and time were studied in order to acquire an appropriate current efficiency of copper deposition. The highest efficiency is about 95% obtaining from electro-winning condition of 3V, 55°C and 3,600 s correspondingly. The cathode copper with 95.5% Cu analyzed using atomic absorption spectrometry can be obtained from this single-winning condition. In order to increase the copper grade, solvent extraction should be used to increase the sulfate concentration, say 50 g/L, prior to winning the cathode copper effectively.

Keywords: copper metal, current efficiency, dilute sulfate solution, electro-winning

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3095 The Biological Function and Clinical Significance of Long Non-coding RNA LINC AC008063 in Head and Neck Squamous Carcinoma

Authors: Maierhaba Mijiti

Abstract:

Objective:The aim is to understand the relationship between the expression level of the long-non-coding RNA LINC AC008063 and the clinicopathological parameters of patients with head and neck squamous cell carcinoma (HNSCC), and to clarify the biological function of LINC AC008063 in HNSCC cells. Moreover, it provides a potential biomarker for the diagnosis, treatment, and prognosis evaluation of HNSCC. Methods: The expression level of LINC AC008063 in the HNSCC was analyzed using transcriptome sequencing data from the TCGA (The cancer genome atlas) database. The expression levels of LINC AC008063 in human embryonic lung diploid cells 2BS, human immortalized keratinocytes HACAT, HNSCC cell lines CAL-27, Detroit562, AMC-HN-8, FD-LSC-1, FaDu and WSU-HN30 were determined by real-time quantitative PCR (qPCR). RNAi (RNA interference) was introduced for LINC AC008063 knockdown in HNSCC cell lines, the localization and abundance analysis of LINC AC008063 was determined by RT-qPCR, and the biological functions were examined by CCK-8, clone formation, flow cytometry, transwell invasion and migration assays, Seahorse assay. Results: LINC AC008063 was upregulated in HNSCC tissue (P<0.001), and verified b CCK-8, clone formation, flow cytometry, transwell invasion and migration assays, Seahorse assayy qPCR in HNSCC cell lines. The survival analysis revealed that the overall survival rate (OS) of patients with high LINC AC008063 expression group was significantly lower than that in the LINC AC008063 expression group, the median survival times for the two groups were 33.10 months and 61.27 months, respectively (P=0.002). The clinical correlation analysis revealed that its expression was positively correlated with the age of patients with HNSCC (P<0.001) and positively correlated with pathological state (T3+T4>T1+T2, P=0.03). The RT-qPCR results showed that LINC AC008063 was mainly enriched in cytoplasm (P=0.01). Knockdown of LINC AC008063 inhibited proliferation, colony formation, migration and invasion; the glycolytic capacity was significantly decreased in HNSCC cell lines (P<0.05). Conclusion: High level of LINC AC008063 was associated with the malignant progression of HNSCC as well as promoting the important biological functions of proliferation, colony formation, migration and invasion; in particular, the glycolytic capacity was decreased in HNSCC cells. Therefore, LINC AC008063 may serve as a potential biomarker for HNSCC and a distinct molecular target to inhibit glycolysis.

Keywords: head and neck squamous cell carcinoma, oncogene, long non-coding RNA, LINC AC008063, invasion and metastasis

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3094 Changed Behavior of the Porcine Hemagglutinating Encephalomyelitis Virus (Betacoronavirus) in Respiratory Epithelial Cells

Authors: Ateeqa Aslam, Hans J. Nauwynck

Abstract:

Porcine hemagglutinating encephalomyelitis virus (PHEV) is a betacoronavirus that has been studied in the past as a cause of vomiting and wasting disease (VWD) in young piglets (<3 weeks). Nowadays, the virus is still circulating on most farms in Belgium, but there are no descriptions anymore of VWD. Therefore, we are interested in differences between the old and new strains. We compared the replication kinetics of the old well-studied strain VW572 (1972) and the recent isolate P412 (2020) in a susceptible continuous cell line (RPD cells) and in primary porcine respiratory epithelial cells (PoRECs). The RPD cell line was inoculated with each PHEV strain at an m.o.i. of 1 the supernatant was collected, and the cells were fixed at different time points post-inoculation. The supernatant was titrated (extracellular virus titer), and the infected cells were revealed by immunofluorescence staining and quantitated by fluorescence microscopy. We found that VW572 replicated better in the RPD cell line at earlier time points when compared to P412. Porcine respiratory epithelial cells (PoREC) were isolated, grown at air-liquid interphase in transwells and inoculated with both strains of PHEV at a virus titer of 106.6TCID50 per 200 µl either at the apical side or at the basal side of the cells. At different time points after inoculation, the transwells were fixed and stained for infected cells. VW572 preferentially infected the epithelial cells via the basolateral side of porcine nasal epithelial cells, whereas P412 preferred the apical side. These findings suggest that there has been an evolution of PHEV in its interaction with the respiratory epithelial cells. In the future, more virus strains will be enclosed and the tropism of the strains for different neuronal cell types will be examined for the change in virus neurotropism.

Keywords: porcine hemagglutinating encephalomyelitis virus (PHEV), primary porcine respiratory epithelial cells (PoRECs), virus tropism, vomiting and wasting disease (VWD)

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3093 Inhibition of Glutamate Carboxypeptidase Activity Protects Retinal Ganglionic Cell Death Induced by Ischemia-Reperfusion by Reducing the Astroglial Activation in Rat

Authors: Dugeree Otgongerel, Kyong Jin Cho, Yu-Han Kim, Sangmee Ahn Jo

Abstract:

Excessive activation of glutamate receptor is thought to be involved in retinal ganglion cell (RGC) death after ischemia- reperfusion damage. Glutamate carboxypeptidase II (GCPII) is an enzyme responsible for the synthesis of glutamate. Several studies showed that inhibition of GCPII prevents or reduces cellular damage in brain diseases. Thus, in this study, we examined the expression of GCPII in rat retina and the role of GCPII in acute high IOP ischemia-reperfusion damage of eye by using a GCPII inhibitor, 2-(phosphonomethyl) pentanedioic acid (2-PMPA). Animal model of ischemia-reperfusion was induced by raising the intraocular pressure for 60 min and followed by reperfusion for 3 days. Rats were randomly divided into four groups: either intra-vitreous injection of 2-PMPA (11 or 110 ng per eye) or PBS after ischemia-reperfusion, 2-PMPA treatment without ischemia-reperfusion and sham-operated normal control. GCPII immunoreactivity in normal rat retina was detected weakly in retinal nerve fiber layer (RNFL) and retinal ganglionic cell layer (RGL) and also inner plexiform layer (IPL) and outer plexiform layer (OPL) strongly where are co-stained with an anti-GFAP antibody, suggesting that GCPII is expressed mostly in Muller and astrocytes. Immunostaining with anti-BRN antibody showed that ischemia- reperfusion caused RGC death (31.5 %) and decreased retinal thickness in all layers of damaged retina, but the treatment of 2-PMPA twice at 0 and 48 hour after reperfusion blocked these retinal damages. GCPII level in RNFL layer was enhanced after ischemia-reperfusion but was blocked by PMPA treatment. This result was confirmed by western blot analysis showing that the level of GCPII protein after ischemia- reperfusion increased by 2.2- fold compared to control, but this increase was blocked almost completely by 110 ng PMPA treatment. Interestingly, GFAP immunoreactivity in the retina after ischemia- reperfusion followed by treatment with PMPA showed similar pattern to GCPII, increase after ischemia-reperfusion but reduction to the normal level by PMPA treatment. Our data demonstrate that increase of GCPII protein level after ischemia-reperfusion injury is likely to cause glial activation and/or retinal cell death which are mediated by glutamate, and GCPII inhibitors may be useful in treatment of retinal disorders in which glutamate excitotoxicity is pathogenic.

Keywords: glutamate carboxypepptidase II, glutamate excitotoxicity, ischemia-reperfusion, retinal ganglion cell

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3092 Sickle Cell Disease: Review of Managements in Pregnancy and the Outcome in Ampang Hospital, Selangor

Authors: Z. Nurzaireena, K. Azalea, T. Azirawaty, S. Jameela, G. Muralitharan

Abstract:

The aim of this study is the review of the management practices of sickle cell disease patients during pregnancy, as well as the maternal and neonatal outcome at Ampang Hospital, Selangor. The study consisted of a review of pregnant patients with sickle cell disease under follow up at the Hematology Clinic, Ampang Hospital over the last seven years to assess their management and maternal-fetal outcome. The results of the review show that Ampang Hospital is considered the public hematology centre for sickle cell disease and had successfully managed three pregnancies throughout the last seven years. Patients’ presentations, managements and maternal-fetal outcome were compared and reviewed for academic improvements. All three patients were seen very early in their pregnancy and had been given a regime of folic acid, antibiotics and thrombo-prophylactic drugs. Close monitoring of maternal and fetal well being was done by the hematologists and obstetricians. Among the patients, there were multiple admissions during the pregnancy for either a painful sickle cell bone crisis, haemolysis following an infection and anemia requiring phenotype- matched blood and exchange transfusions. Broad spectrum antibiotics coverage during and infection, hydration, pain management and venous-thrombolism prophylaxis were mandatory. The pregnancies managed to reach near term in the third trimester but all required emergency caesarean section for obstetric indications. All pregnancies resulted in live births with good fetal outcome. During post partum all were nursed closely in the high dependency units for further complications and were discharged well. Post partum follow up and contraception counseling was comprehensively given for future pregnancies. Sickle cell disease is uncommonly seen in the East, especially in the South East Asian region, yet more cases are seen in the current decade due to improved medical expertise and advance medical laboratory technologies. Pregnancy itself is a risk factor for sickle cell patients as increased thrombosis event and risk of infections can lead to multiple crisis, haemolysis, anemia and vaso-occlusive complications including eclampsia, cerebrovasular accidents and acute bone pain. Patients mostly require multiple blood product transfusions thus phenotype-matched blood is required to reduce the risk of alloimmunozation. Emphasizing the risks and complications in preconception counseling and establishing an ultimate pregnancy plan would probably reduce the risk of morbidity and mortality to the mother and unborn child. Early management for risk of infection, thromboembolic events and adequate hydration is mandatory. A holistic approach involving multidisciplinary team care between the hematologist, obstetricians, anesthetist, neonatologist and close nursing care for both mother and baby would ensure the best outcome. In conclusion, sickle cell disease by itself is a high risk medical condition and pregnancy would further amplify the risk. Thus, close monitoring with combine multidisciplinary care, counseling and educating the patients are crucial in achieving the safe outcome.

Keywords: anaemia, haemoglobinopathies, pregnancy, sickle cell disease

Procedia PDF Downloads 258
3091 Modified 'Perturb and Observe' with 'Incremental Conductance' Algorithm for Maximum Power Point Tracking

Authors: H. Fuad Usman, M. Rafay Khan Sial, Shahzaib Hamid

Abstract:

The trend of renewable energy resources has been amplified due to global warming and other environmental related complications in the 21st century. Recent research has very much emphasized on the generation of electrical power through renewable resources like solar, wind, hydro, geothermal, etc. The use of the photovoltaic cell has become very public as it is very useful for the domestic and commercial purpose overall the world. Although a single cell gives the low voltage output but connecting a number of cells in a series formed a complete module of the photovoltaic cells, it is becoming a financial investment as the use of it fetching popular. This also reduced the prices of the photovoltaic cell which gives the customers a confident of using this source for their electrical use. Photovoltaic cell gives the MPPT at single specific point of operation at a given temperature and level of solar intensity received at a given surface whereas the focal point changes over a large range depending upon the manufacturing factor, temperature conditions, intensity for insolation, instantaneous conditions for shading and aging factor for the photovoltaic cells. Two improved algorithms have been proposed in this article for the MPPT. The widely used algorithms are the ‘Incremental Conductance’ and ‘Perturb and Observe’ algorithms. To extract the maximum power from the source to the load, the duty cycle of the convertor will be effectively controlled. After assessing the previous techniques, this paper presents the improved and reformed idea of harvesting maximum power point from the photovoltaic cells. A thoroughly go through of the previous ideas has been observed before constructing the improvement in the traditional technique of MPP. Each technique has its own importance and boundaries at various weather conditions. An improved technique of implementing the use of both ‘Perturb and Observe’ and ‘Incremental Conductance’ is introduced.

Keywords: duty cycle, MPPT (Maximum Power Point Tracking), perturb and observe (P&O), photovoltaic module

Procedia PDF Downloads 176
3090 Clinicopathological and Immunohistochemical Study of Ovarian Sex Cord-Stromal Tumors and Their Histological Mimics

Authors: Ghada Esheba, Ebtisam Aljerayan, Afnan Al-Ghamdi, Atheer Alsharif, Hanan alzahrani

Abstract:

Background: Primary ovarian neoplasms comprise a heterogeneous group of tumors of three main subtypes: surface epithelial, germ cell, and sex cord-stromal. The wide morphological variation within and between these groups can result in diagnostic difficulties. Gonadal sex cord-stromal tumors (SCST) represent one of the most heterogeneous categories of human neoplasms, because they may contain various combinations of different gonadal sex cord and stromal element. Aim: The aim of this work is to highlight the clinicopathological characteristics of SCST and to assess the value of alpha-inhibin and calretinin in the distinction between SCST and their mimics. Material and methods: This study was carried out on 100 cases using full tissue sections; 70 cases were SCST and 30 cases were histological mimics of SCST. The cases were studied using immunohistochemically using alpha-inhibin. In addition, an ovarian tissue microarray containing 170 benign and malignant ovarian neoplasms was also studied immunohistochemically for calretinin expression. The ovarian microarray included 14 SCST, 59 ovarian serous borderline tumors, 17 mucinous borderline tumors, 10 mucinous adenocarcinomas, 32 endometrioid adenocarcinomas, 34 clear cell carcinomas, and 4 germ cell tumors. Results: 99% of SCST examined using full tissue sections exhibited positive cytoplasmic staining for inhibin. On the contrary, only 7% of the histological mimics (P value < 0.0001). 86% of SCST in the tissue microarray were positive for calretinin with nuclear and/or cytoplasmic staining compared to only 7% of the other tumor types (P value < 0.0001). Conclusions: SCST have characteristic clinicopathological and immunohistochemical features and their recognition is crucial for proper diagnosis and treatment. Alpha-inhibin and calretinin are of great help in the diagnosis of sex cord-stromal tumors.

Keywords: calretinin, granulosa cell tumor, inhibin, sex cord-stromal tumors

Procedia PDF Downloads 208
3089 Effect of Diindolylmethane on BBN-Induced Bladder Carcinogenesis in Rats

Authors: Sundaresan Sivapatham, B. Prabhu

Abstract:

Cancer results from a multistage, multi-mechanism carcinogenesis process that involves mutagenic, cell death and epigenetic mechanisms, during the three distinguishable but closely allied stages: initiation, promotion, and progression. Chemoprevention is promising in the realm of cancer prevention and it has been shown to reduce the risk of development of carcinoma in highly susceptible individuals such as those with known genetic mutations or high level of risk factors. The present study is aimed at the need of early detection of bladder cancer in order to improve performance in the treatment of this disease. Consumption of certain natural products like DIM is associated with a reduction in cancer incidence in humans. The study showed the protective effects of Diindolylmethane in N-Butyl-N-(4-hydroxybutyl) nitrosamine treated rats. Results of the study had shown the changes in the tumor markers, biomarkers and histopathological alterations in experimental rats when compared to control rats. The protective effects of DIM were shown from the results of cell proliferation, apoptotic markers and histopathological findings when compared with experimental control animals. Hence, our results speculate that the tumor markers, apoptotic markers, histopathological changes and cell proliferation index measured as PCNA serves as an indicator suggestive of protective effects of DIM in BBN induced urinary bladder carcinogenesis.

Keywords: bladder cancer, N-Butyl-N-(4-hydroxybutyl) nitrosamine, diindolylmethane, histopathology

Procedia PDF Downloads 342
3088 Biocompatible Chitosan Nanoparticles as an Efficient Delivery Vehicle for Mycobacterium Tuberculosis Lipids to Induce Potent Cytokines and Antibody Response through Activation of γδ T-Cells in Mice

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

Abstract:

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

Procedia PDF Downloads 280
3087 Chemical Analysis and Cytotoxic Evaluation of Asphodelus Aestivus Brot. Flowers

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

Abstract:

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

Procedia PDF Downloads 222
3086 Rapid Start-Up and Efficient Long-Term Nitritation of Low Strength Ammonium Wastewater with a Sequencing Batch Reactor Containing Immobilized Cells

Authors: Hammad Khan, Wookeun Bae

Abstract:

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

Procedia PDF Downloads 273
3085 Effects of Anti-FGL2 Monoclonal Antibody SPF89 on Vascular Inflammation

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

Abstract:

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

Procedia PDF Downloads 271
3084 Method of Visual Prosthesis Design Based on Biologically Inspired Design

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

Abstract:

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

Procedia PDF Downloads 192
3083 Revolutionizing Mobility: Decoding Electric Vehicles (EVs) and Hydrogen Fuel Cell Vehicles (HFCVs)

Authors: Samarjeet Singh, Shubhank Arya, Shubham Chauhan

Abstract:

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

Procedia PDF Downloads 45
3082 Study of Pressure and Air Mass Flow Effect on Output Power of PEM Fuel Cell Powertrains in Vehicles and Airplanes- A Simulation-based Approach

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

Abstract:

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

Procedia PDF Downloads 175
3081 Assessment of Cellular Metabolites and Impedance for Early Diagnosis of Oral Cancer among Habitual Smokers

Authors: Ripon Sarkar, Kabita Chaterjee, Ananya Barui

Abstract:

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

Procedia PDF Downloads 176