Search results for: cell stimulation
3525 Developing Scaffolds for Tissue Regeneration using Low Temperature Plasma (LTP)
Authors: Komal Vig
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Cardiovascular disease (CVD)-related deaths occur in 17.3 million people globally each year, accounting for 30% of all deaths worldwide, with a predicted annual incidence of deaths to reach 23.3 million globally by 2030. Autologous bypass grafts remain an important therapeutic option for the treatment of CVD, but the poor quality of the donor patient’s blood vessels, the invasiveness of the resection surgery, and postoperative movement restrictions create issues. The present study is aimed to improve the endothelialization of intimal surface of graft by using low temperature plasma (LTP) to increase the cell attachment and proliferation. Polytetrafluoroethylene (PTFE) was treated with LTP. Air was used as the feed-gas, and the pressure in the plasma chamber was kept at 800 mTorr. Scaffolds were also modified with gelatin and collagen by dipping method. Human umbilical vein endothelial cells (HUVEC) were plated on the developed scaffolds, and cell proliferation was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and by microscopy. mRNA expressions levels of different cell markers were investigated using quantitative real-time PCR (qPCR). XPS confirmed the introduction of oxygenated functionalities from LTP. HUVEC cells showed 80% seeding efficiency on the scaffold. Microscopic and MTT assays indicated increase in cell viability in LTP treated scaffolds, especially when treated with gelatin or collagen, compared to untreated scaffolds. Gene expression studies shows enhanced expression of cell adhesion marker Integrin- α 5 gene after LTP treatment. LTP treated scaffolds exhibited better cell proliferation and viability compared to untreated scaffolds. Protein treatment of scaffold increased cell proliferation. Based on our initial results, more scaffolds alternatives will be developed and investigated for cell growth and vascularization studies. Acknowledgments: This work is supported by the NSF EPSCoR RII-Track-1 Cooperative Agreement OIA-2148653.Keywords: LTP, HUVEC cells, vascular graft, endothelialization
Procedia PDF Downloads 643524 Sirt1 Activators Promote Skin Cell Regeneration and Cutaneous Wound Healing
Authors: Hussain Mustatab Wahedi, Sun You Kim
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Skin acts as a barrier against the harmful environmental factors. Integrity and timely recovery of the skin from injuries and harmful effects of radiations is thus very important. This study aimed to investigate the importance of Sirt1 in the recovery of skin from UVB-induced damage and cutaneous wounds by using natural and synthetic novel Sirt1 activators. Juglone, known as a natural Pin1 inhibitor, and NED416 a novel synthetic Sirt1 activator were checked for their ability to regulate the expression and activity of Sirt1 and hence photo-damage and wound healing in cultured skin cells (NHDF and HaCaT cells) and mouse model by using Sirt1 siRNA knockdown, cell migration assay, GST-Pulldown assay, western blot analysis, tube formation assay, and immunohistochemistry. Interestingly, Sirt1 knockdown inhibited skin cell migration in vitro. Juglone up regulated the expression of Sirt1 in both the cell lines under normal and UVB irradiated conditions, enhanced Sirt1 activity and increased the cell viability by reducing reactive oxygen species synthesis and apoptosis. Juglone promoted wound healing by increasing cell migration and angiogenesis through Cdc42/Rac1/PAK, MAPKs and Smad pathways in skin cells. NED416 upregulated Sirt1 expression in HaCaT and NHDF cells as well as increased Sirt1 activity. NED416 promoted the process of wound healing in early as well as later stages by increasing macrophage recruitment, skin cell migration, and angiogenesis through Cdc42/Rac1 and MAPKs pathways. So, both these compounds activated Sirt1 and promoted the process of wound healing thus pointing towards the possible role of Sirt1 in skin regeneration and wound healing.Keywords: skin regeneration, wound healing, Sirt1, UVB light
Procedia PDF Downloads 1843523 Optimum Design for Cathode Microstructure of Solid Oxide Fuel Cell
Authors: M. Riazat, H. Abdolvand, M. Baniassadi
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In this present work, 3D reconstruction of cathode of SOFC is developed with various volume fractions and porosity. Three Phase Boundary (TPB) of construction of such derived micro structures is calculated. The neural network is used to optimize the porosity and volume fraction of each phase to reach a structure with maximum TPB.Keywords: fuel cell, solid oxide, TPB, 3D reconstruction
Procedia PDF Downloads 3173522 SOCS1 Inhibits MDR1 in Mammary Cell Carcinoma Reverses Multidrug Resistance
Authors: Debasish Pradhan, Shaktiprasad Pradhan, Rakesh Kumar Pradhan, Gitanjali Tripathy
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Suppressors of cytokine signalling (SOCS1), a newly indentified antiapoptotic molecule is a downstream effector of the receptor tyrosine kinase-Ras signalling pathway. The current study has uncovered that SOCS1 may have wide and imperative capacities, particularly because of its close correlation with malignant tumors. To investigate the impact of SOCS1 on MDR, we analyzed the expression of P-gp and SOCS1 by immunohistochemistry and found there was a positive correlation between them. At that point, we effectively interfered with RNA translation by the contamination of siRNA of SOCS1 into MCF7/ADM breast cancer cell lines through a lentivirus, and the expression of the target gene was significantly inhibited. After RNAi, the drug resistance was reduced altogether and the expression of MDR1 mRNA and P-gp in MCF7/ADM cell lines demonstrated a significant decrease. Likewise, the expression of P53 protein increased in a statistically significant manner (p ≤ 0.01) after RNAi exposure. Moreover, flow cytometry analysis uncovers that cell cycle and anti-apoptotic enhancing capacity of cells changed after RNAi treatment. These outcomes proposed SOCS1 may take part in breast cancer MDR by managing MDR1 and P53 expression, changing cell cycle and enhancing the anti-apoptotic ability.Keywords: breast cancer, multidrug resistance, SOCS1 gene, MDR1 gene, RNA interference
Procedia PDF Downloads 3523521 Feasibility of Leukemia Cancer Treatment (K562) by Atmospheric Pressure Plasma Jet
Authors: Mashayekh Amir Shahriar, Akhlaghi Morteza, Rajaee Hajar, Khani Mohammad Reza, Shokri Babak
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A new and novel approach in medicine is the use of cold plasma for various applications such as sterilization blood coagulation and cancer cell treatment. In this paper a pin-to-hole plasma jet suitable for biological applications is investigated, characterized and the possibility and feasibility of cancer cell treatment is evaluated. The characterization includes power consumption via Lissajous method, thermal behavior of plasma using Infra-red camera as a novel method, Optical Emission Spectroscopy (OES) to determine the species that are generated. Treatment of leukemia cancer cells is also implemented and MTT assay is used to evaluate viability.Keywords: Atmospheric Pressure Plasma Jet (APPJ), Plasma Medicine, Cancer cell treatment, leukemia, Optical Emission
Procedia PDF Downloads 6503520 Annona muricata Leaves Induced Mitochondrial-Mediated Apoptosis in A549 Cells
Authors: Soheil Zorofchian Moghadamtousi, Habsah Abdul Kadir, Mohammadjavad Paydar, Elham Rouhollahi, Hamed Karimian
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The present study was designed to evaluate the molecular mechanisms of Annona muricata leaves ethyl acetate extract (AMEAE) against lung cancer A549 cells. Cell viability analysis revealed the selective cytotoxic effect of AMEAE towards A549 cells. Treatment of A549 cells with AMEAE significantly elevated the reactive oxygen species formation, followed by attenuation of mitochondrial membrane potential via upregulation of Bax and downregulation of Bcl-2, accompanied by cytochrome c release to the cytosol. The released cytochrome c triggered the activation of caspase-9 followed by caspase-3. In addition, AMEAE-induced apoptosis was accompanied by cell cycle arrest at G1 phase. Our data showed for the first time that AMEAE inhibited the proliferation of A549 cells, leading to cell cycle arrest and programmed cell death through activation of the mitochondrial-mediated signaling pathway.Keywords: Annona muricata, lung cancer, apoptosis, mitochondria
Procedia PDF Downloads 3143519 Viscoelastic Cell Concentration in a High Aspect Ratio Microchannel Using a Non-Powered Air Compressor
Authors: Jeonghun Nam, Seonggil Kim, Hyunjoo Choi, Chae Seung Lim
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Quantification and analysis of rare cells are challenging in clinical applications and cell biology due to its extremely small number in blood. In this work, we propose a viscoelastic microfluidic device for continuous cell concentration without sheath flows. Due to the viscoelastic effect on suspending cells, cells with the blockage ratio higher than 0.1 could be tightly focused at the center of the microchannel. The blockage ratio was defined as the particle diameter divided by the channel width. Finally, cells were concentrated through the center outlet and the additional suspending medium was removed to the side outlets. Since viscoelastic focusing is insensitive to the flow rate higher than 10 μl/min, the non-powered hand pump sprayer could be used with no accurate control of the flow rate, which is suitable for clinical settings in resource-limited developing countries. Using multiple concentration processes, high-throughput concentration of white blood cells in lysed blood sample was achieved by ~ 300-fold.Keywords: cell concentration, high-throughput, non-powered, viscoelastic fluid
Procedia PDF Downloads 2843518 ZVZCT PWM Boost DC-DC Converter
Authors: Ismail Aksoy, Haci Bodur, Nihan Altintaş
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This paper introduces a boost converter with a new active snubber cell. In this circuit, all of the semiconductor components in the converter softly turns on and turns off with the help of the active snubber cell. Compared to the other converters, the proposed converter has advantages of size, number of components and cost. The main feature of proposed converter is that the extra voltage stresses do not occur on the main switches and main diodes. Also, the current stress on the main switch is acceptable level. Moreover, the proposed converter can operates under light load conditions and wide input line voltage. In this study, the operating principle of the proposed converter is presented and its operation is verified with the Proteus simulation software for a 1 kW and 100 kHz model.Keywords: active snubber cell, boost converter, zero current switching, zero voltage switching
Procedia PDF Downloads 10223517 Omni-Relay (OR) Scheme-Aided LTE-A Communication Systems
Authors: Hassan Mahasneh, Abu Sesay
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We propose the use of relay terminals at the cell edge of an LTE-based cellar system. Each relay terminal is equipped with an omni-directional antenna. We refer to this scheme as the Omni-Relay (OR) scheme. The OR scheme coordinates the inter-cell interference (ICI) stemming from adjacent cells and increases the desired signal level at cell-edge regions. To validate the performance of the OR scheme, we derive the average signal-to-interference plus noise ratio (SINR) and the average capacity and compare it with the conventional universal frequency reuse factor (UFRF). The results show that the proposed OR scheme provides higher average SINR and average capacity compared to the UFRF due to the assistance of the distributed relay nodes.Keywords: the UFRF scheme, the OR scheme, ICI, relay terminals, SINR, spectral efficiency
Procedia PDF Downloads 3353516 Analysis of BSF Layer N-Gaas/P-Gaas/P+-Gaas Solar Cell
Authors: Abderrahmane Hemmani, Hamid Khachab, Dennai Benmoussa, Hassane Benslimane, Abderrachid Helmaoui
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Back surface field GaAs with n -p-p+ structures are found to have better characteristics than the conventional solar cells. A theory, based on the transport of both minority carriers under the charge neutrality condition, has been developed in the present paper which explains behavior of the back surface field solar cells. That is reported with an efficiency of 25,05% (Jsc=33.5mA/cm2, Vco=0.87v and fill factor 86% under AM1.5 global conditions). We present the effect of technological parameters of the p+ layer on the conversion efficiency on the solar cell. Good agreement is achieved between our results and the simulation results given the variation of the equivalent recombination velocity to p+ layer as a function of BSF thickness and BSF doping.Keywords: back surface field, GaAs, solar cell, technological parameters
Procedia PDF Downloads 4283515 The Effect of Education given to Parents of Children with Sickle Cell Anemia in Turkey and Chad to Reduce Children's Pain
Authors: Fatima El Zahra Amin, Emine Efe
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This study was carried out to evaluate the effect of the education program for parents of children with Sickle Cell Anemia, on the knowledge level of parents and the reduction of pain relief by non-pharmacological methods used by parents at home. In Turkey, 54 parents and 109 from Chad agreed to participate in the survey. The data were collected by the researcher using a face-to-face interview method. Non-pharmacological treatment information form for parents, face expressions rating scale, and parent education program for non-pharmacological methods used in children with sickle cell anemia were used. It was determined that there was a statistically significant difference between the educational status, occupation, disease status, place of residence, family structure and age of parents of Chad and Turkey. According to the ratings of facial expressions scale, it was concluded that there was no significant difference between the children’s average degree of pain before and after administration of non-pharmacological methods by the groups of Chad and Turkey. It was determined that the educational programs prepared for parents of children with sickle cell anemia in both Turkey and Chad were effective in increasing the knowledge level of parents and also in reducing pain crisis with non-pharmacological methods parents used at home.Keywords: Chad, child, non-pharmacological treatment methods, nurse, sickle cell anemia, Turkey
Procedia PDF Downloads 2613514 Music Therapy Intervention as a Means of Stimulating Communicative Abilities of Seniors with Neurocognitive Disorders – Theory versus Practice
Authors: Pavel Svoboda, Oldřich Müller
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The paper contains a screening of the opinions of helping professional workers working in a home for seniors with individuals with neurocognitive disorders and compares them with the opinions of a younger generation of students who are just preparing for this work. The authors carried out a comparative questionnaire survey with both target groups, focusing on the analysis and comparison of possible differences in their knowledge in the field of care for elderly people with neurocognitive disorders. Specifically, they focused on knowledge and experience with approaches, methods and tools applicable within the framework of music therapy interventions, as they are understood in practice in comparison with the theoretical knowledge of secondary school students focused on social work. The questionnaire was mainly aimed at assessing the knowledge of the possibilities of effective memory stimulation of the elderly and their communication skills using the means of music. The conducted investigation was based on the research of studies dealing with so-called non-pharmacological approaches to the given clientele; for professional caregivers, it followed music therapy lessons, which the authors regularly implemented from the beginning of 2022. Its results will, among other things, serve as the basis for an upcoming study with a scoping design review.Keywords: neurocognitive disorders, seniors, music therapy intervention, melody, rhythm, text, memory stimulation, communication skills
Procedia PDF Downloads 663513 Studies on Induction of Cytotoxicity Through Apoptosis In Ovarian Cancer Cell Line (CAOV-3) by Chloroform Extract of Artocarpus Kemando Miq
Authors: Noor Shafifiyaz Mohd Yazid, Najihah Mohd Hashim, Hapipah Mohd Ali, Syam Mohan, Rosea Go
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Artocarpus kemando is a plant species from Moraceae family. This plant is used as household utensil by the local and the fruits are edible. The plants’ bark was used for the extraction process and yielded the chloroform crude extract which was used to screen for anticancer potential. The cytotoxic effect of the extract on CAOV-3 and WRL 68 cell lines were determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide or MTT assays. Qualitative AO/PI assay was performed to confirm the apoptosis and necrosis process. Meanwhile, the measurement of cell loss, nuclear morphology, DNA content, cell membrane permeability, mitochondrial membrane potential changes and cytochrome c release from mitochondria were detected through cytotoxicity 3 assay. In MTT assay, A. kemando inhibited 50% growth of CAOV-3 cells at 27.9 ± 0:03, 20.1± 0:03, 18.21± 0:04 µg/mL after 24, 48 and 72 hour, respectively. The morphology changes can be seen on CAOV-3 with a production of cell membrane blebbing, cromatin condensation and apoptotic bodies. Evaluation of cytotoxicity 3 on CAOV-3 cells after treated with extract resulting loss of mitochondrial membrane potential and release of cytochrome c from mitochondria. The results demonstrated A. kemando has potentially anticancer agent, particularly on human ovarian cancer.Keywords: anticancer, Artocarpus kemando, ovarian cancer, cytotoxicity
Procedia PDF Downloads 5433512 The Effect of Combined Doxorubicin and Dioscorea esculenta on Apoptosis Induction in Human Breast Cancer Cells
Authors: Dina Fatmawati, Sofia Mubarika, Mae Sri Wahyuningsih
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Chemotherapy for breast cancer is largely ineffective, but innovative combinations of chemotherapeutic agents and natural compounds represent a promising strategy. In our previous study, the combination of Doxorubicin (Dox) and ethanolic extract of Dioscorea esculenta tuber ((EED) was found to have a synergistic effect on T47D human breast cancer cell line. In this study, we investigated the apoptotic effect of the combination on T47D human breast cancer cells and normal fibroblasts cell line and its effects on the expression of Caspase-3 and cleaved poly (ADP-Ribose) Polymerase-1 (cPARP-1) protein. T47D cell lines and fibroblasts cells were treated with the combination of Dox and EED. Apoptotic effect of the combination was determined using flow cytrometry assay. Protein expressions were determined by immunocytochemistry staining. The percentage of apoptotic cells were significantly higher in T47D cell lines (75%) than that of in fibroblast cells (23%). The expression of Caspase 3 (84.53%) and cPARP-1 (83.36%) were significantly higher in the cancer cell lines than those of normal cells. These results indicate that the combination of doxorubicin and Dioscorea esculenta is a promising candidate for the treatment of breast cancer cells.Keywords: Dioscorea esculenta, Doxorubicin, apoptosis, immunocytochemistry, cancer cells
Procedia PDF Downloads 4463511 The Amount of Organic Phosphates (Like DPG) Existing in Blood is Determining Factor of Mammal’s Bulk
Authors: Ramin Amirmardfar
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Throughout Necessary oxygen should be supplied for all cells of a mammal at any moment through blood to make it possible remain alive all cells the mammal’s body. In case a mammal’s bulk is large, there is a farther distance between cells in different tissues and mammals’ heart. Therefore red blood cells in bulky mammal’s body should be capable of conveying oxygen to farther distances. To make it practical, oxygen should be glued red blood cells tenaciously. In other words, cohesion strength of oxygen to red blood cell of bulky mammal’s blood should be much more than the same of small mammal’s blood. In mammal’s bodies, the controlling factor of amount of cohesion of oxygen to red blood cell, are organic phosphates (like DPG). The less DPG in red blood cells of a mammal, the more cohesion of oxygen to red blood cell (at the same rate). As much as oxygen is glued more tenacious to red blood cells, oxygen could been carried to farther distance and as much as oxygen could be conveyed to farther points of heart, bulk of mammal could be larger at the same rate.Keywords: mammals size, animals size, organic phosphates, DPG, red blood cell, metabolism
Procedia PDF Downloads 3513510 Influence of Cucurbitacin-Containing Phytonematicides on Growth of Rough Lemon (Citrus jambhiri)
Authors: Raisibe V. Mathabatha, Phatu W. Mashela, Nehemiah M. Mokgalong
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Occasional incidence of phytotoxicity in Nemarioc-BL and Nemafric-AL phytonematicides to crops raises credibility challenges that could negate their registration as commercial products. Responses of plants to phytonematicides are characterized by the existence of stimulation, neutral and inhibition phases, with the mid-point of the former being referred to as the Mean Concentration Stimulation Point (MSCP = Dm + Rh/2). The objective of this study was to determine the MCSP and the overall sensitivity (∑k) of Nemarioc-AL and Nemafric-BL phytonematicides to rough lemon seedling rootstocks using the Curve-fitting Allelochemical Response Dosage (CARD) computer-based model. Two parallel greenhouse experiments were initiated, with seven dilutions of each phytonematicide arranged in a randomised complete block design, replicated nine times. Six-month-old rough lemon seedlings were transplanted into 20-cm-diameter plastic pots, filled with steam-pasteurised river sand (300°C for 3 h) and Hygromix-T growing mixture. Treatments at 0, 2, 4, 8, 16, 32 and 164% dilutions were applied weekly at 300 ml/plant. At 84 days after the treatments, analysis of variance-significant plant variables was subjected to the CARD model to generate appropriate biological indices. Computed MCSP values for Nemarioc-AL and Nemafric-BL phytonematicides on rough lemon were 29 and 38%, respectively, whereas ∑k values were 1 and 0, respectively. At the applied concentrations, rough lemon seedlings were highly sensitive to Nemarioc-AL and Nemafric-BL phytonematicides.Keywords: crude extracts, cucurbitacins, effective microbes, fruit extracts
Procedia PDF Downloads 1413509 Harnessing of Electricity from Distillery Effluent and Simultaneous Effluent Treatment by Microbial Fuel Cell
Authors: Hanish Mohammed, C. H. Muthukumar Muthuchamy
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The advancement in the science and technology has made it possible to convert electrical energy into any desired form. It has given electrical energy a place of pride in the modern world. The survival of industrial undertakings and our social structure depends primarily upon low cost and uninterrupted supply of electrical energy. Microbial fuel cell (MFC) is a promising and emerging technique for sustainable bioelectricity generation and wastewater treatment. MFCs are devices which are capable of converting organic matter to electricity/hydrogen with help of microorganisms. Different kinds of wastewater could be used in this technique, distillery effluent is one of the most troublesome and complex and strong organic effluent with high chemical oxygen demand of 1,53,846 mg/L. A single cell MFC unit was designed and fabricated for the distillery effluent treatment and to generate electricity. Due to the high COD value of the distillery effluent helped in the production of energy for 74 days. The highest voltage got from the fuel cell is 206 mV on the 30th day. A maximum power density obtained from the MFC was 9.8 mW, treatment efficiency was evaluated in terms of COD removal and other parameters. COD removal efficiencies were around 68.5 % and other parameters such as Total Hardness (81.5%), turbidity (70 %), chloride (66%), phosphate (79.5%), Nitrate (77%) and sulphate (71%). MFC using distillery effluent is a promising new unexplored substrate for the power generation and sustainable treatment technique through harnessing of bioelectricity.Keywords: microbial fuel cell (MFC), bioelectricity, distillery effluent, wastewater treatment
Procedia PDF Downloads 2053508 Distracted Driving among Young Drivers in Qatar
Authors: Khaled Shaaban
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Distracted driving, which includes anything that distracts a driver from the main task of driving, is one of the main causes of traffic accidents in modern societies. The objective of this research was to understand the type of activities that young drivers perform while driving in Qatar and to identify which activities cause the most distraction to the driver based on their experience. The data was collected through administered questionnaires in the city of Doha, Qatar. According to the participants, the majority reported that they use their cell phone all the time or occasionally while driving. Other significantly cited activities while driving included listening to music or radio, talking with passengers, and eating, drinking or smoking. When asked about the activities that distract the driver, using cell phone was listed as the most distracting activity followed by mental activities and adjusting GPS and audio device vehicle.Keywords: driver distraction, young drivers, cell phone use, Qatar
Procedia PDF Downloads 4853507 Synthesis and Antiproliferative Activity of 5-Phenyl-N3-(4-fluorophenyl)-4H-1,2,4-triazole-3,4-diamine Derivatives
Authors: L. Mallesha, P. Mallu, B. Veeresh
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In the present study, 2, 6-diflurobenzohydrazide and 4-fluorophenylisothiocyanate were used as the starting materials to synthesize 5-phenyl-N3-(4-fluorophenyl)-4H-1, 2, 4-triazole-3, 4-diamine. Further, compound 5-phenyl-N3-(4-fluorophenyl)-4H-1, 2, 4-triazole-3,4-diamine reacted with fluoro substituted benzaldehydes to yield a series of Schiff bases. All the final compounds were characterized using IR, 1H NMR, 13C NMR, MS and elemental analyses. New compounds were evaluated for their antiproliferative effect using the MTT assay method against four human cancer cell lines (K562, COLO-205, MDA-MB231, and IMR-32) for the time period of 24 h. Among the series, few compounds showed good activity on all cell lines, whereas the other compounds in the series exhibited moderate activity.Keywords: Schiff bases, MTT assay, antiproliferative activity, human cancer cell lines, 1, 2, 4-triazoles
Procedia PDF Downloads 3673506 A Single Cell Omics Experiments as Tool for Benchmarking Bioinformatics Oncology Data Analysis Tools
Authors: Maddalena Arigoni, Maria Luisa Ratto, Raffaele A. Calogero, Luca Alessandri
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The presence of tumor heterogeneity, where distinct cancer cells exhibit diverse morphological and phenotypic profiles, including gene expression, metabolism, and proliferation, poses challenges for molecular prognostic markers and patient classification for targeted therapies. Understanding the causes and progression of cancer requires research efforts aimed at characterizing heterogeneity, which can be facilitated by evolving single-cell sequencing technologies. However, analyzing single-cell data necessitates computational methods that often lack objective validation. Therefore, the establishment of benchmarking datasets is necessary to provide a controlled environment for validating bioinformatics tools in the field of single-cell oncology. Benchmarking bioinformatics tools for single-cell experiments can be costly due to the high expense involved. Therefore, datasets used for benchmarking are typically sourced from publicly available experiments, which often lack a comprehensive cell annotation. This limitation can affect the accuracy and effectiveness of such experiments as benchmarking tools. To address this issue, we introduce omics benchmark experiments designed to evaluate bioinformatics tools to depict the heterogeneity in single-cell tumor experiments. We conducted single-cell RNA sequencing on six lung cancer tumor cell lines that display resistant clones upon treatment of EGFR mutated tumors and are characterized by driver genes, namely ROS1, ALK, HER2, MET, KRAS, and BRAF. These driver genes are associated with downstream networks controlled by EGFR mutations, such as JAK-STAT, PI3K-AKT-mTOR, and MEK-ERK. The experiment also featured an EGFR-mutated cell line. Using 10XGenomics platform with cellplex technology, we analyzed the seven cell lines together with a pseudo-immunological microenvironment consisting of PBMC cells labeled with the Biolegend TotalSeq™-B Human Universal Cocktail (CITEseq). This technology allowed for independent labeling of each cell line and single-cell analysis of the pooled seven cell lines and the pseudo-microenvironment. The data generated from the aforementioned experiments are available as part of an online tool, which allows users to define cell heterogeneity and generates count tables as an output. The tool provides the cell line derivation for each cell and cell annotations for the pseudo-microenvironment based on CITEseq data by an experienced immunologist. Additionally, we created a range of pseudo-tumor tissues using different ratios of the aforementioned cells embedded in matrigel. These tissues were analyzed using 10XGenomics (FFPE samples) and Curio Bioscience (fresh frozen samples) platforms for spatial transcriptomics, further expanding the scope of our benchmark experiments. The benchmark experiments we conducted provide a unique opportunity to evaluate the performance of bioinformatics tools for detecting and characterizing tumor heterogeneity at the single-cell level. Overall, our experiments provide a controlled and standardized environment for assessing the accuracy and robustness of bioinformatics tools for studying tumor heterogeneity at the single-cell level, which can ultimately lead to more precise and effective cancer diagnosis and treatment.Keywords: single cell omics, benchmark, spatial transcriptomics, CITEseq
Procedia PDF Downloads 1063505 The Study of Blood Consumption for Stem Cell Transplant Patients in Shahid Ghazi Tabatabaei Hospital, Tabriz, Iran
Authors: Naser Shagerdi Esmaeli, Mohsen Hamidpour, Parisa Hasankhani Tehrani
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Background And Objective: Haematopoietic stem cell transplant is a potentially curative treatment option in various benign and malignant haematological diseases. Patients undergoing stem cell transplant procedure require blood transfusion on a daily basis. Currently, there is paucity of data from developing countries on transfusion practices. This audit was undertaken to determine the consumption of packed red blood cells (PRBCs) transfusion in the bone marrow transplant unit of the Shahid Ghazi Tabatabaei Hospital, Tabriz, Iran. Subjects And Methods: A retrospective audit was conducted for packed red cell transfusion ordering practice over a period from March 2017 to march 2018. All consecutive patients admitted for stem cell transplant procedure for various underlying diseases were included. Outcome measures used in this study were (i) cross match to transfusion (C: T) ratio and (ii) transfusion trigger. Results: During the study period, n=13 patients underwent a haematopoietic stem cell transplant. There were n=10 males and n=3 females. One patient was less than 15 years of age, while rests were adults. Median age±SD was 26.5±14.5 years (12∼54 years). The underlying diagnosis included Aplastic anemia (n=4), Thalassemia major (n=1), Multiple Myeloma (n=3), Acute leukemia (n=3), Hodgkin's lymphoma (n=1), PRCA (n=1). Grand total consumption of PRBCs during the study period was 204, while 258 products were crossmatch. The C:T ratio was 1.26. The transfusion trigger was Hb level of less than 8 gr/dl. Conclusion: The results of our BMT unit indicate that the C:T ratio and transfusion trigger is comparable to the international criteria and pioneer country in BMT transplantation. Also, we hope that our blood consumption become less than it is now.Keywords: blood consumption, C: T ratio, PRBCs, stem cell transplant, tabriz, Iran
Procedia PDF Downloads 1183504 The Effects of Metformin And PCL-sorafenib Nanoparticles Co-treatment on MCF-7 Cell Culture Model of Breast Cancer
Authors: Emad Heydarnia, Aref Sepasi, Nika Asefi, Sara Khakshournia, Javad Mohammadnejad
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Background: Despite breakthrough therapeutics in breast cancer, it is one of the main causes of mortality among women worldwide. Thus, drug therapies for treating breast cancer have recently been developed by scientists. Metformin and Sorafenib are well-known therapeutic in breast cancer. In the present study, we combined Sorafenib and PCL-sorafenib with metformin to improve drug absorption and promote therapeutic efficiency. Methods: The MCF-7 cells were treated with Metformin, Sorafenib, or PCL-sorafenib. The growth inhibitory effect of these drugs and cell viability were assessed using MTT and flow cytometry assays, respectively. The expression of targeted genes involved in cell proliferation, signaling, and the cell cycle was measured by Real-time PCR. Results: The results showed that MCF-7 cells treated with Metformin/Sorafenib and PCL-sorafenib/Metformin co-treatment contributed to 50% viability compared to untreated group. Moreover, PI and Annexin V staining tests showed that the cells viability for Metformin/Sorafenib and PCL-sorafenib/Metformin was 38% and 17%, respectively. Furthermore, Sorafenib/Metformin and PCL-sorafenib/Metformin leads to p53 gene expression increase by which they can increase ROS, thereby decreasing GPX4 gene expression. In addition, they affected the expression of BCL2, and BAX genes and altered the cell cycle. Conclusion: Together, the combination of PCL-sorafenib/Metformin and Sorafenib/Metformin increased Sorafenib absorption at lower doses and also leads to apoptosis and oxidative stress increases in MCF-7 cells.Keywords: breast cancer, metformin, nanotechnology, sorafenib
Procedia PDF Downloads 633503 PNIPAAm-MAA Nanoparticles as Delivery Vehicles for Curcumin Against MCF-7 Breast Cancer Cells
Authors: H. Tayefih, F. farajzade ahari, F. Zarghami, V. Zeighamian, N. Zarghami, Y. Pilehvar-soltanahmadi
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Breast cancer is the most frequently occurring cancer among women throughout the world. Natural compounds such as curcumin hold promise to treat a variety of cancers including breast cancer. However, curcumin's therapeutic application is limited, due to its rapid degradation and poor aqueous solubility. On the other hand, previous studies have stated that drug delivery using nanoparticles might improve the therapeutic response to anticancer drugs. Poly (N-isopropylacrylamide-co-methacrylic acid) (PNIPAAm–MAA) is one of the hydrogel copolymers utilized in the drug delivery system for cancer therapy. The aim of this study was to examine the cytotoxic potential of curcumin encapsulated within the NIPAAm-MAA nanoparticle, on the MCF-7 breast cancer cell line. In this work, polymeric nanoparticles were synthesized through the free radical mechanism, and curcumin was encapsulated into NIPAAm-MAA nanoparticles. Then, the cytotoxic effect of curcumin-loaded NIPAAm-MAA on the MCF-7 breast cancer cell line was measured by MTT assays. The evaluation of the results showed that curcumin-loaded NIPAAm-MAA has more cytotoxic effect on the MCF-7 cell line and efficiently inhibited the growth of the breast cancer cell population, compared with free curcumin. In conclusion, this study indicates that curcumin-loaded NIPAAm-MAA suppresses the growth of the MCF-7 cell line. Overall, it is concluded that encapsulating curcumin into the NIPAAm-MAA copolymer could open up new avenues for breast cancer treatment.Keywords: PNIPAAm-MAA, breast cancer, curcumin, drug delivery
Procedia PDF Downloads 3693502 Based on MR Spectroscopy, Metabolite Ratio Analysis of MRI Images for Metastatic Lesion
Authors: Hossain A, Hossain S.
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Introduction: In a small cohort, we sought to assess the magnetic resonance spectroscopy's (MRS) ability to predict the presence of metastatic lesions. Method: A Popular Diagnostic Centre Limited enrolled patients with neuroepithelial tumors. The 1H CSI MRS of the brain allows us to detect changes in the concentration of specific metabolites caused by metastatic lesions. Among these metabolites are N-acetyl-aspartate (NNA), creatine (Cr), and choline (Cho). For Cho, NAA, Cr, and Cr₂, the metabolic ratio was calculated using the division method. Results: The NAA values were 0.63 and 5.65 for tumor cells, 1.86 and 5.66 for normal cells, and 1.86 and 5.66 for normal cells 2. NAA values for normal cells 1 were 1.84, 10.6, and 1.86 for normal cells 2, respectively. Cho levels were as low as 0.8 and 10.53 in the tumor cell, compared to 1.12 and 2.7 in the normal cell 1 and 1.24 and 6.36 in the normal cell 2. Cho/Cr₂ barely distinguished itself from the other ratios in terms of significance. For tumor cells, the ratios of Cho/NAA, Cho/Cr₂, NAA/Cho, and NAA/Cr₂ were significant. Normal cell 1 had significant Cho/NAA, Cho/Cr, NAA/Cho, and NAA/Cr ratios. Conclusion: The clinical result can be improved by using 1H-MRSI to guide the size of resection for metastatic lesions. Even though it is non-invasive and doesn't present any difficulties during the procedure, MRS has been shown to predict the detection of metastatic lesions.Keywords: metabolite ratio, MRI images, metastatic lesion, MR spectroscopy, N-acetyl-aspartate
Procedia PDF Downloads 893501 Level of Awareness of Genetic Counselling in Benue State Nigeria: Its Advocacy on the Inheritance of Sickle Cell Disease
Authors: Agi Sunday
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A descriptive analysis of reported cases of sickle cell disease and the level of awareness about genetic counselling in 30 hospitals were carried out. Additionally, 150 individuals between ages 16-45 were randomly selected for evaluation of genetic counselling awareness. The main tools for this study were questionnaires which were taken to hospitals, and individuals completed the others. The numbers of reported cases of sickle cell disease recorded in private, public and teaching hospitals were 14 and 57; 143 and 89; 272 and 57 for the periods of 1995-2000 and 2001-2005, respectively. A general informal genetic counselling took place mostly in the hospitals visited. 122 (86%) individuals had the knowledge of genetic disease and only 43 (30.3%) individuals have been exposed to genetic counselling. 64% of individuals agreed that genetic counselling would help in the prevention of genetic disease.Keywords: sickle disease, genetic counseling, genetic testing, advocacy
Procedia PDF Downloads 3833500 Chemical Composition of Essential Oil and in vitro Antibacterial and Anticancer Activity of the Hydroalcolic Extract from Coronilla varia
Authors: A. A. Dehpour, B. Eslami, S. Rezaie, S. F. Hashemian, F. Shafie, M. Kiaie
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The aims of study were investigation on chemical composition essential oil and the effect of extract of Coronilla varia on antimicrobial and cytotoxicity activity. The essential oils of Coronilla varia is obtained by hydrodistillation and analyzed by (GC/MS) for determining their chemical composition and identification of their components. Antibacterial activity of plant extract was determined by disc diffusion method. The effect of hydroalcolic extracts from Cornilla varia investigated on MCF7 cancer cell line by MTT assay. The major components were Caryophyllene Oxide (60.19%), Alphacadinol (4.13%) and Homoadantaneca Robexylic Acid (3.31%). The extracts from Coronilla varia had interesting activity against Proteus mirabilis in the concentration of 700 µg/disc and did not show any activity against Staphylococus aureus, Bacillus subtillis, Klebsiella pneumonia and Entrobacter cloacae. The positive control, Ampicillin, Chloramphenicol and Cenphalothin had shown zone of inhibition resistant all bacteria. Corohilla varia ethanol extract could inhibit the proliferation of MCF7 cell line in RPMI 1640 medium. IC50 5(mg/ml) was the optimum concentration of extract from Coronilla varia inhibition of cell line growth. The MCF7 cancer cell line and Proteus mirabilis were more sensitive to Coronilla varia ethanol extract.Keywords: Coronilla varia, essential oil, antibacterial, anticancer, hela cell line
Procedia PDF Downloads 3863499 Evaluation of Anticancer and Antioxidant Activity of Purified Lovastatin from Aspergillus terreus (KM017963)
Authors: Bhargavi Santebennur Dwarakanath, Praveen Vadakke Kamath, Savitha Janakiraman
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Cervical cancer is one of the leading causes of mortality in women and is the second most common malignancy worldwide. Lovastatin, a non polar, anticholesterol drug which also exerts antitumour activity in vitro. In the present study, lovastatin from Aspergillus terreus (KM017963) was purified by adsoprtion chromatography and evaluated for its anticancer and anti-oxidant properties in human cervical cancer cell lines (HeLa). The growth inhibitory and proapoptotic effects of purified lovastatin on HeLa cell lines were investigated by determining its influence on cytotoxicity, Mitochondrial Membrane Potential (MMP), DNA fragmentation and antioxidant property (Hydroxy radical scavenging effect and the levels of total reduced glutathione). Flow cytometry analysis by propidium iodide staining confirmed the induction of apoptotic cell death and revealed cell cycle arrest at G0/G1 phase. Results of the study give leads for anticancer effects of lovastatin and its potential efficacy in the chemotherapy of cervical cancer.Keywords: apoptosis, Aspergillus terreus, cervical cancer, lovastatin
Procedia PDF Downloads 3013498 The Effects of Highly Active Antiretroviral Therapy (HAART) on the Expression of Muc1 and P65 in a Cervical Cancer Cell Line, HCS-2
Authors: K. R. Thabethe, G. A. Adefolaju, M. J. Hosie
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Cervical cancer is the third most commonly diagnosed cancer globally and it is one of three AIDS defining malignancies. Highly active antiretroviral therapy (HAART) is a combination of three or more antiretroviral drugs and has been shown to play a significant role in reducing the incidence of some AIDS defining malignancies, although its effect on cervical cancer is still unclear. The aim of this study was to investigate the relationship between cervical cancer and HAART. This was achieved by studying the expression of two signalling molecules expressed in cervical cancer; MUC1 and P65. Following the 24 hour treatment of a cervical cancer cell line, HCS-2, with drugs which are commonly used as part of HAART at their clinical plasma concentrations, real-time qPCR and immunofluorescence were used in order to study gene and protein expression. A one way ANOVA followed by a Tukey Kramer Post Hoc test was conducted using JMP 11 software on both sets of data. The drug classified as a protease inhibitor (PI) (i.e. LPV/r) reduced MUC1 and P65 gene and protein expression more than the other drug tested. PIs are known to play a significant role in cell death, therefore the cells were thought to be more susceptible to cell death following treatment with PIs. In conclusion, the drugs used, especially the PI showed some anticancer effects by facilitating cell death through decreased gene and protein expression of MUC1 and P65 and present promising agents for cancer treatment.Keywords: cervical cancer, haart, MUC1, P65
Procedia PDF Downloads 3263497 Physiological Normoxia and Cellular Adhesion of Diffuse Large B-Cell Lymphoma Primary Cells: Real-Time PCR and Immunohistochemistry Study
Authors: Kamila Duś-Szachniewicz, Kinga M. Walaszek, Paweł Skiba, Paweł Kołodziej, Piotr Ziółkowski
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Cell adhesion is of fundamental importance in the cell communication, signaling, and motility, and its dysfunction occurs prevalently during cancer progression. The knowledge of the molecular and cellular processes involved in abnormalities in cancer cells adhesion has greatly increased, and it has been focused mainly on cellular adhesion molecules (CAMs) and tumor microenvironment. Unfortunately, most of the data regarding CAMs expression relates to study on cells maintained in standard oxygen condition of 21%, while the emerging evidence suggests that culturing cells in ambient air is far from physiological. In fact, oxygen in human tissues ranges from 1 to 11%. The aim of this study was to compare the effects of physiological lymph node normoxia (5% O2), and hyperoxia (21% O2) on the expression of cellular adhesion molecules of primary diffuse large B-cell lymphoma cells (DLBCL) isolated from 10 lymphoma patients. Quantitative RT-PCR and immunohistochemistry were used to confirm the differential expression of several CAMs, including ICAM, CD83, CD81, CD44, depending on the level of oxygen. Our findings also suggest that DLBCL cells maintained at ambient O2 (21%) exhibit reduced growth rate and migration ability compared to the cells growing in normoxia conditions. Taking into account all the observations, we emphasize the need to identify the optimal human cell culture conditions mimicking the physiological aspects of tumor growth and differentiation.Keywords: adhesion molecules, diffuse large B-cell lymphoma, physiological normoxia, quantitative RT-PCR
Procedia PDF Downloads 2713496 Derivation of Human NK Cells from T Cell-Derived Induced Pluripotent Stem Cells Using Xenogeneic Serum-Free and Feeder Cell-Free Culture System
Authors: Aliya Sekenova, Vyacheslav Ogay
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The derivation of human induced pluripotent stem cells (iPSCs) from somatic cells by direct reprogramming opens wide perspectives in the regenerative medicine. It means the possibility to develop the personal and, consequently, any immunologically compatible cells for applications in cell-based therapy. The purpose of our study was to develop the technology for the production of NK cells from T cell-derived induced pluripotent stem cells (TiPSCs) for subsequent application in adoptive cancer immunotherapy. Methods: In this study iPSCs were derived from peripheral blood T cells using Sendai virus vectors expressing Oct4, Sox2, Klf4 and c-Myc. Pluripotent characteristics of TiPSCs were examined and confirmed with alkaline phosphatase staining, immunocytochemistry and RT-PCR analysis. For NK cell differentiation, embryoid bodies (EB) formed from (TiPSCs) were cultured in xenogeneic serum-free medium containing human serum, IL-3, IL-7, IL-15, SCF, FLT3L without using M210-B4 and AFT-024 stromal feeder cells. After differentiation, NK cells were characterized with immunofluorescence analysis, flow cytometry and cytotoxicity assay. Results: Here, we for the first time demonstrate that TiPSCs can effectively differentiate into functionally active NK cells without M210-B4 and AFT-024 xenogeneic stroma cells. Immunofluorescence and flow cytometry analysis showed that EB-derived cells can differentiate into a homogeneous population of NK cell expressing high levels of CD56, CD45 and CD16 specific markers. Moreover, these cells significantly express killing activation receptors such as NKp44 and NKp46. In the comparative analysis, we observed that NK cells derived using feeder-free culture system have more high killing activity against K-562 tumor cells, than NK cells derived by feeder-dependent method. Thus, we think that our obtained data will be useful for the development of large-scale production of NK cells for translation into cancer immunotherapy.Keywords: induced pluripotent stem cells, NK cells, T cells, cell diffentiation, feeder cell-free culture system
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