Search results for: primary limbal stromal cells
7504 The Role of Okra (Abelmoschus esculentus Linn.) on Lipopolysaccharide-Induced Reactive Oxygen Species and Inflammatory Mediator in BV2 Microglial Cells
Authors: Nootchanat Mairuae, Walaiporn Tongjaroenbuangam, Chalisa Louicharoen Cheepsunthorn, Poonlarp Cheepsunthorn
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The aim of this study was to investigate the anti-oxidative effect, the anti-inflammatory effects, and the molecular mechanisms of okra (Abelmoschus esculentus Linn.) on lipopolysaccharide (LPS)-stimulated BV2 microglial cells. The BV2 cells were treated with LPS in the presence or absence of okra. Reactive oxygen species (ROS) and nitric oxide (NO) production were measured using the ROS detection reagent DCF-DA and the Griess reaction, respectively. The phosphorylation levels of nuclear factor-kappa B (NF-kB) p65 was detected by Western blot assay. Treatment of BV2 microglia cells with okra was found to significantly suppress the LPS-induced inflammatory mediator NO as well as ROS compared to untreated cells. The levels of LPS-induced NF-kB p65 phosphorylation were significantly decreased following okra treatment too. These results show that okra exerts anti-oxidative and anti-inflammatory effects in LPS-stimulated BV2 microglial cells by suppressing the NF-κB pathway. This suggests okra might be a valuable agent for treatment of anti-neuroinflammatory diseases mediated by microglial cells.Keywords: Abelmoschus esculentus Linn, microglia, neuroinflammation, reactive oxygen spicy
Procedia PDF Downloads 2957503 Neighbour Cell List Reduction in Multi-Tier Heterogeneous Networks
Authors: Mohanad Alhabo, Naveed Nawaz
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The ongoing call or data session must be maintained to ensure a good quality of service. This can be accomplished by performing the handover procedure while the user is on the move. However, the dense deployment of small cells in 5G networks is a challenging issue due to the extensive number of handovers. In this paper, a neighbour cell list method is proposed to reduce the number of target small cells and hence minimizing the number of handovers. The neighbour cell list is built by omitting cells that could cause an unnecessary handover and handover failure because of short time of stay of the user in these cells. A multi-attribute decision making technique, simple additive weighting, is then applied to the optimized neighbour cell list. Multi-tier small cells network is considered in this work. The performance of the proposed method is analysed and compared with that of the existing methods. Results disclose that our method has decreased the candidate small cell list, unnecessary handovers, handover failure, and short time of stay cells compared to the competitive method.Keywords: handover, HetNets, multi-attribute decision making, small cells
Procedia PDF Downloads 1247502 Immunoliposomes Conjugated with CD133 Antibody for Targeting Melanoma Cancer Stem Cells
Authors: Chuan Yin
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Cancer stem cells (CSCs) represent a subpopulation of cancer cells that possess the characteristics associated with normal stem cells. CD133 is a phenotype of melanoma CSCs responsible for melanoma metastasis and drug resistance. Although adriamycin (ADR) is commonly used drug in melanoma therapy, but it is ineffective in the treatment of melanoma CSCs. In this study, we constructed CD133 antibody conjugated ADR immunoliposomes (ADR-Lip-CD133) to target CD133+ melanoma CSCs. The results showed that the immunoliposomes possessed a small particle size (~150 nm), high drug encapsulation efficiency (~90%). After 72 hr treatment on the WM266-4 melanoma tumorspheres, the IC50 values of the drug formulated in ADR-Lip-CD133, ADR-Lip (ADR liposomes) and ADR are found to be 24.42, 57.13 and 59.98 ng/ml respectively, suggesting that ADR-Lip-CD133 was more effective than ADR-Lip and ADR. Significantly, ADR-Lip-CD133 could almost completely abolish the tumorigenic ability of WM266-4 tumorspheres in vivo, and showed the best therapeutic effect in WM266-4 melanoma xenograft mice. It is noteworthy that ADR-Lip-CD133 could selectively kill CD133+ melanoma CSCs of WM266-4 cells both in vitro and in vivo. ADR-Lip-CD133 represent a potential approach in targeting and killing CD133+ melanoma CSCs.Keywords: cancer stem cells, melanoma, immunoliposomes, CD133
Procedia PDF Downloads 3887501 An Improved Circulating Tumor Cells Analysis Method for Identifying Tumorous Blood Cells
Authors: Salvador Garcia Bernal, Chi Zheng, Keqi Zhang, Lei Mao
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Circulating Tumor Cells (CTC) is used to detect tumoral cell metastases using blood samples of patients with cancer (lung, breast, etc.). Using an immunofluorescent method a three channel image (Red, Green, and Blue) are obtained. These set of images usually overpass the 11 x 30 M pixels in size. An aided tool is designed for imaging cell analysis to segmented and identify the tumorous cell based on the three markers signals. Our Method, it is cell-based (area and cell shape) considering each channel information and extracting and making decisions if it is a valid CTC. The system also gives information about number and size of tumor cells found in the sample. We present results in real-life samples achieving acceptable performance in identifying CTCs in short time.Keywords: Circulating Tumor Cells (CTC), cell analysis, immunofluorescent, medical image analysis
Procedia PDF Downloads 2187500 Co-Culture of Neonate Mouse Spermatogonial Stem Cells with Sertoli Cells: Inductive Role of Melatonin following Transplantation: Adult Azoospermia Mouse Model
Authors: Mehdi Abbasi, Shadan Navid, Mohammad Pourahmadi, M. Majidi Zolbin
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We have recently reported that melatonin as antioxidant enhances the efficacy of colonization of spermatogonial stem cells (SSCs). Melatonin as an antioxidant plays a vital role in the development of SSCs in vitro. This study aimed to investigate evaluation of sertoli cells and melatonin simultaneously on SSC proliferation following transplantation to testis of adult mouse busulfan-treated azoospermia model. SSCs and sertoli cells were isolated from the testes of three to six-day old male mice.To determine the purity, Flow cytometry technique using PLZF antibody were evaluated. Isolated testicular cells were cultured in αMEM medium in the absence (control group) or presence (experimental group) of sertoli cells and melatonin extract for 2 weeks. We then transplanted SSCs by injection into the azoospermia mice model. Higher viability, proliferation, and Id4, Plzf, expression were observed in the presence of simultaneous sertoli cells and melatonin in vitro. Moreover, immunocytochemistry results showed higher Oct4 expression in this group. Eight weeks after transplantation, injected cells were localized at the base of seminiferous tubules in the recipient testes. The number of spermatogonia and the weight of testis were higher in the experimental group relative to control group. The results of our study suggest that this new protocol can increase the transplantation of these cells can be useful in the treatment of male infertility.Keywords: colonization, melatonin, spermatogonial stem cell, transplantation
Procedia PDF Downloads 1737499 Comparison of Transparent Nickel Doped Cobalt Sulfide and Platinum Counter Electrodes Used in Quasi-Solid State Dye Sensitized Solar Cells
Authors: Dimitra Sygkridou, Dimitrios Karageorgopoulos, Elias Stathatos, Evangelos Vitoratos
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Transparent nickel doped cobalt sulfide was fabricated on a SnO2:F electrode and tested as an efficient electrocatalyst and as an alternative to the expensive platinum counter electrode. In order to investigate how this electrode could affect the electrical characteristics of a dye-sensitized solar cell, we manufactured cells with the same TiO2 photoanode sensitized with dye (N719) and employing the same quasi-solid electrolyte, altering only the counter electrode used. The cells were electrically and electrochemically characterized and it was observed that the ones with the Ni doped CoS2 outperformed the efficiency of the cells with the Pt counter electrode (3.76% and 3.44% respectively). Particularly, the higher efficiency of the cells with the Ni doped CoS2 counter electrode (CE) is mainly because of the enhanced photocurrent density which is attributed to the enhanced electrocatalytic ability of the CE and the low charge transfer resistance at the CE/electrolyte interface.Keywords: nickel doped cobalt sulfide, counter electrodes, dye-sensitized solar cells, quasi-solid state electrolyte, hybrid organic-inorganic materials
Procedia PDF Downloads 7637498 Performance and Lifetime of Tandem Organic Solar Cells
Authors: Guillaume Schuchardt, Solenn Berson, Gerard Perrier
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Multi-junction solar cell configurations, where two sub-cells with complementary absorption are stacked and connected in series, offer an exciting approach to tackle the single junction limitations of organic solar cells and improve their power conversion efficiency. However, the augmentation of the number of layers has, as a consequence, to increase the risk of reducing the lifetime of the cell due to the ageing phenomena present at the interfaces. In this work, we study the intrinsic degradation mechanisms, under continuous illumination AM1.5G, inert atmosphere and room temperature, in single and tandem organic solar cells using Impedance Spectroscopy, IV Curves, External Quantum Efficiency, Steady-State Photocarrier Grating, Scanning Kelvin Probe and UV-Visible light.Keywords: single and tandem organic solar cells, intrinsic degradation mechanisms, characterization: SKP, EQE, SSPG, UV-Visible, Impedance Spectroscopy, optical simulation
Procedia PDF Downloads 3667497 Malignant Idiopathic Intracranial Hypertension Revealed a Hidden Primary Spinal Leptomeningeal Medulloblastoma
Authors: Naim Izet Kajtazi
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Context: Frequently, the cause of raised intracranial pressure remains unresolved and rarely is related to spinal tumors, moreover less to spinal medulloblastoma without primary brain focus. Process: An 18-year-old woman had a 3-month history of headaches and impaired vision. Neurological examination revealed bilateral sixth cranial nerve palsies with bilateral papilloedema of grade III. No focal brain or spine lesion was found on imaging. Consecutive lumbar punctures showed high opening pressure and subsequent increasing protein level. The meningeal biopsy was negative. At one point, she developed an increasing headache, vomiting and back pain. Spine MRI showed diffuse nodular leptomeningeal enhancement with the largest nodule at T6–T7. Malignant cells were detected in cerebrospinal fluid. She underwent laminectomy with excisional biopsy, and pathology showed medulloblastoma WHO grade IV. Outcome: She was treated with chemotherapy and craniospinal irradiation and made a good recovery. Relevance: Primary spinal leptomeningeal medulloblastoma is extremely rare, especially without primary brain focus, but may cause increased intracranial pressure, even in the early microscopic phases, and it should be considered in the differential diagnosis if conventional and aggressive treatment of idiopathic intracranial hypertension fails. We assume that arachnoiditis from tumor seeding caused increased intracranial pressure. Appropriate neurosurgical intervention and surgical biopsy are mandated if a suspicious lesion is detected. Consider proper rescreening of the whole neuroaxis in refractory cases of intracranial hypertension.Keywords: CNS infection, IIH, headache, primary spinal leptomeningeal medulloblastoma
Procedia PDF Downloads 697496 Removal of Samarium in Environmental Water Samples by Modified Yeast Cells
Authors: Homayon Ahmad Panahi, Seyed Mehdi Seyed Nejad, Elham Moniri
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A novel bio-adsorbent is fabricated by attaching a cibacron blue to yeast cells. The modified bio-sorbent has been characterized by some techniques like Fourier transform infrared spectroscopy (FT-IR) and elemental analysis (CHN) and applied for the preconcentration and determination of samarium from aqueous water samples. The best pH value for adsorption of the brilliant crecyle blue by yeast cells- cibacron blue was 7. The sorption capacity of modified biosorbent was 18.5 mg. g⁻¹. A recovery of 95.3% was obtained for Sm(III) when eluted with 0.5 M nitric acid. The method was applied for Sm(III) preconcentration and determination in river water sample.Keywords: samarium, solid phase extraction, yeast cells, water sample, removal
Procedia PDF Downloads 2637495 Genetic Association of SIX6 Gene with Pathogenesis of Glaucoma
Authors: Riffat Iqbal, Sidra Ihsan, Andleeb Batool, Maryam Mukhtar
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Glaucoma is a gathering of optic neuropathies described by dynamic degeneration of retinal ganglionic cells. It is clinically and innately heterogenous illness containing a couple of particular forms each with various causes and severities. Primary open-angle glaucoma (POAG) is the most generally perceived kind of glaucoma. This study investigated the genetic association of single nucleotide polymorphisms (SNPs; rs10483727 and rs33912345) at the SIX1/SIX6 locus with primary open-angle glaucoma (POAG) in the Pakistani population. The SIX6 gene plays an important role in ocular development and has been associated with morphology of the optic nerve. A total of 100 patients clinically diagnosed with glaucoma and 100 control individuals of age over 40 were enrolled in the study. Genomic DNA was extracted by organic extraction method. The SNP genotyping was done by (i) PCR based restriction fragment length polymorphism (RFLP) and sequencing method. Significant genetic associations were observed for rs10483727 (risk allele T) and rs33912345 (risk allele C) with POAG. Hence, it was concluded that Six6 gene is genetically associated with pathogenesis of Glaucoma in Pakistan.Keywords: genotyping, Pakistani population, primary open-angle glaucoma, SIX6 gene
Procedia PDF Downloads 1897494 Viscoelastic Separation and Concentration of Candida Using a Low Aspect Ratio Microchannel
Authors: Seonggil Kim, Jeonghun Nam, Chae Seung Lim
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Rapid diagnosis of fungal infections is critical for rapid antifungal therapy. However, it is difficult to detect extremely low concentration fungi in blood sample. To address the limitation, separation and concentration of fungi in blood sample are required to enhance the sensitivity of PCR analysis. In this study, we demonstrated a sheathless separation and concentration of fungi, candida cells using a viscoelastic fluid. To validate the performance of the device, microparticle mixture (2 and 13 μm) was used, and those particles were successfully separated based on the size difference at high flow rate of 100 μl/min. For the final application, successful separation of the Candida cells from the white blood cells (WBCs) was achieved. Based on the viscoelastic lateral migration toward the equilibrium position, Candida cells were separated and concentrated by center focusing, while WBCs were removed by patterning into two streams between the channel center and the sidewalls. By flow cytometric analysis, the separation efficiency and the purity were evaluated as ~99% and ~ 97%, respectively. From the results, the device can be the powerful tool for detecting extremely rare disease-related cells.Keywords: candida cells, concentration, separation, viscoelastic fluid
Procedia PDF Downloads 1987493 Increased Cytolytic Activity of Effector T-Cells against Cholangiocarcinoma Cells by Self-Differentiated Dendritic Cells with Down-Regulation of Interleukin-10 and Transforming Growth Factor-β Receptors
Authors: Chutamas Thepmalee, Aussara Panya, Mutita Junking, Jatuporn Sujjitjoon, Nunghathai Sawasdee, Pa-Thai Yenchitsomanus
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Cholangiocarcinoma (CCA) is an aggressive malignancy of bile duct epithelial cells in which the standard treatments, including surgery, radiotherapy, chemotherapy, and targeted therapy are partially effective. Many solid tumors including CCA escape host immune responses by creating tumor microenvironment and generating immunosuppressive cytokines such as interleukin-10 (IL-10) and transforming growth factor-β (TGF-β). These cytokines can inhibit dendritic cell (DC) differentiation and function, leading to decreased activation and response of effector CD4+ and CD8+ T cells for cancer cell elimination. To overcome the effects of these immunosuppressive cytokines and to increase ability of DC to activate effector CD4+ and CD8+ T cells, we generated self-differentiated DCs (SD-DCs) with down-regulation of IL-10 and TGF-β receptors for activation of effector CD4+ and CD8+ T cells. Human peripheral blood monocytes were initially transduced with lentiviral particles containing the genes encoding GM-CSF and IL-4 and then secondly transduced with lentiviral particles containing short-hairpin RNAs (shRNAs) to knock-down mRNAs of IL-10 and TGF-β receptors. The generated SD-DCs showed up-regulation of MHC class II (HLA-DR) and co-stimulatory molecules (CD40 and CD86), comparable to those of DCs generated by convention method. Suppression of IL-10 and TGF-β receptors on SD-DCs by specific shRNAs significantly increased levels of IFN-γ and also increased cytolytic activity of DC-activated effector T cells against CCA cell lines (KKU-213 and KKU-100), but it had little effect to immortalized cholangiocytes (MMNK-1). Thus, SD-DCs with down-regulation of IL-10 and TGF-β receptors increased activation of effector T cells, which is a recommended method to improve DC function for the preparation of DC-activated effector T cells for adoptive T-cell therapy.Keywords: cholangiocarcinoma, IL-10 receptor, self-differentiated dendritic cells, TGF-β receptor
Procedia PDF Downloads 1447492 Modeling of Processes Running in Radical Clusters Formed by Ionizing Radiation with the Help of Continuous Petri Nets and Oxygen Effect
Authors: J. Barilla, M. Lokajíček, H. Pisaková, P. Simr
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The final biological effect of ionizing particles may be influenced strongly by some chemical substances present in cells mainly in the case of low-LET radiation. The influence of oxygen may be particularly important because oxygen is always present in living cells. The corresponding processes are then running mainly in the chemical stage of radio biological mechanism. The radical clusters formed by densely ionizing ends of primary or secondary charged particles are mainly responsible for final biological effect. The damage effect depends then on radical concentration at a time when the cluster meets a DNA molecule. It may be strongly influenced by oxygen present in a cell as oxygen may act in different directions: at small concentration of it the interaction with hydrogen radicals prevails while at higher concentrations additional efficient oxygen radicals may be formed. The basic radical concentration in individual clusters diminishes, which is influenced by two parallel processes: chemical reactions and diffusion of corresponding clusters. The given simultaneous evolution may be modeled and analyzed well with the help of Continuous Petri nets. The influence of other substances present in cells during irradiation may be studied, too. Some results concerning the impact of oxygen content will be presented.Keywords: radiobiological mechanism, chemical phase, DSB formation, Petri nets
Procedia PDF Downloads 3167491 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 1027490 Evaluation of Gene Expression after in Vitro Differentiation of Human Bone Marrow-Derived Stem Cells to Insulin-Producing Cells
Authors: Mahmoud M. Zakaria, Omnia F. Elmoursi, Mahmoud M. Gabr, Camelia A. AbdelMalak, Mohamed A. Ghoneim
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Many protocols were publicized for differentiation of human mesenchymal stem cells (MSCS) into insulin-producing cells (IPCs) in order to excrete insulin hormone ingoing to treat diabetes disease. Our aim is to evaluate relative gene expression for each independent protocol. Human bone marrow cells were derived from three volunteers that suffer diabetes disease. After expansion of mesenchymal stem cells, differentiation of these cells was done by three different protocols (the one-step protocol was used conophylline protein, the two steps protocol was depending on trichostatin-A, and the three-step protocol was started by beta-mercaptoethanol). Evaluation of gene expression was carried out by real-time PCR: Pancreatic endocrine genes, transcription factors, glucose transporter, precursor markers, pancreatic enzymes, proteolytic cleavage, extracellular matrix and cell surface protein. Quantitation of insulin secretion was detected by immunofluorescence technique in 24-well plate. Most of the genes studied were up-regulated in the in vitro differentiated cells, and also insulin production was observed in the three independent protocols. There were some slight increases in expression of endocrine mRNA of two-step protocol and its insulin production. So, the two-step protocol was showed a more efficient in expressing of pancreatic endocrine genes and its insulin production than the other two protocols.Keywords: mesenchymal stem cells, insulin producing cells, conophylline protein, trichostatin-A, beta-mercaptoethanol, gene expression, immunofluorescence technique
Procedia PDF Downloads 2217489 A Novel Application of CORDYCEPIN (Cordycepssinensis Extract): Maintaining Stem Cell Pluripotency and Improving iPS Generation Efficiency
Authors: Shih-Ping Liu, Cheng-Hsuan Chang, Yu-Chuen Huang, Shih-Yin Chen, Woei-Cherng Shyu
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Embryonic stem cells (ES) and induced pluripotnet stem cells (iPS) are both pluripotent stem cells. For mouse stem cells culture technology, leukemia inhibitory factor (LIF) was used to maintain the pluripotency of stem cells in vitro. However, LIF is an expensive reagent. The goal of this study was to find out a pure compound extracted from Chinese herbal medicine that could maintain stem cells pluripotency to replace LIF and improve the iPS generation efficiency. From 20 candidates traditional Chinese medicine we found that Cordycepsmilitaris triggered the up-regulation of stem cells activating genes (Oct4 and Sox2) expression levels in MEF cells. Cordycepin, a major active component of Cordycepsmilitaris, also could up-regulate Oct4 and Sox2 gene expression. Furthermore, we used ES and iPS cells and treated them with different concentrations of Cordycepin (replaced LIF in the culture medium) to test whether it was useful to maintain the pluripotency. The results showed higher expression levels of several stem cells markers in 10 μM Cordycepin-treated ES and iPS cells compared to controls that did not contain LIF, including alkaline phosphatase, SSEA1, and Nanog. Embryonic body formation and differentiation confirmed that 10 μM Cordycepin-containing medium was capable to maintain stem cells pluripotency after four times passages. For mechanism analysis, microarray analysis indicated extracellular matrix and Jak/Stat signaling pathway as the top two deregulated pathways. In ECM pathway, we determined that the integrin αVβ5 expression levels and phosphorylated Src levels increased after Cordycepin treatment. In addition, the phosphorylated Jak2 and phosphorylated Sat3 protein levels were increased after Cordycepin treatment and suppressed with the Jak2 inhibitor, AG490. The expression of cytokines associated with Jak2/Stat3 signaling pathway were also up-regulated by Q-PCR and ELISA assay. Lastly, we used Oct4-GFP MEF cells to test iPS generation efficiency following Cordycepin treatment. We observed that 10 Μm Cordycepin significantly increased the iPS generation efficiency in day 21. In conclusion, we demonstrated Cordycepin could maintain the pluripotency of stem cells through both of ECM and Jak2/Stat3 signaling pathway and improved iPS generation efficiency.Keywords: cordycepin, iPS cells, Jak2/Stat3 signaling pathway, molecular biology
Procedia PDF Downloads 4437488 Chemical Modification of Biosorbent for Prconcentation of Cadmium in Water Sample
Authors: Homayon Ahmad Panahi, Niusha Mohseni Darabi, Elham Moniri
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A new biosorbent is prepared by coupling a cibacron blue to yeast cells. The modified yeast cells with cibacron blue has been characterized by Fourier transform infrared spectroscopy (FT-IR) and elemental analysis and applied for the preconcentration and solid phase extraction of trace cadmium ion from water samples. The optimum pH value for sorption of the cadmium ions by yeast cells- cibacron blue was 5.5. The sorption capacity of modified biosorbent was 45 mg. g−1. A recovery of 98.2% was obtained for Cd(II) when eluted with 0.5 M nitric acid. The method was applied for Cd(II) preconcentration and determination in sea water sample.Keywords: solid phase extraction, yeast cells, Nickl, isotherm study
Procedia PDF Downloads 2687487 The Role of Il-6-Mediated NS5ATP9 Expression in Autophagy of Liver Cancer Cells
Authors: Hongping Lu, Kelbinur Tursun, Yaru Li, Yu Zhang, Shunai Liu, Ming Han
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Objective: To investigate whether NS5ATP9 is involved in IL-6 mediated autophagy and the relationship between IL-6 and NS5ATP9 in liver cancer cells. Methods: 1. Detect the mRNA and protein levels of Beclin 1 after HepG2 cells were treated with or without recombinant human IL-6 protein. 2. Measure and compare of the changes of autophagy-related genes with their respective control, after IL-6 was silenced or neutralized with monoclonal antibody against human IL-6. 3. HepG2 cells were incubated with 50 ng/ml of IL-6 in the presence or absence of PDTC. The expression of NS5ATP9 was analyzed by Western blot after 48 h. 4. After NS5ATP9-silenced HepG2 cells had been treated with 50 ng/ml recombinant IL-6 protein, we detected the Beclin 1 and LC3B (LC3Ⅱ/Ⅰ) expression. 5. HepG2 cells were transfected with pNS5ATP9, si-NS5ATP9, and their respective control. Total RNA was isolated from cells and analyzed for IL-6. 6. Silence or neutralization of IL-6 in HepG2 cells which has been transfected with NS5ATP9. Beclin 1 and LC3 protein levels were analyzed by Western blot. Result: 1. After HepG2 were treated with recombinant human IL-6 protein, the expression of endogenous Beclin 1 was up-regulated at mRNA and protein level, and the conversion of endogenous LC3-I to LC3-II was also increased. These results indicated that IL-6 could induce autophagy. 2. When HepG2 cells were treated with IL-6 siRNA or monoclonal antibody against human IL-6, the expression of autophagy-related genes were decreased. 3. Exogenous human IL-6 recombinant protein up-regulated NS5ATP9 via NF-κB activation. 4. The expression of Beclin 1 and LC3B was down-regulated after IL-6 treated NS5ATP9-silenced HepG2 cells. 5. NS5ATP9 could reverse regulates IL-6 expression in HepG2 cells. 6. Silence or neutralization of IL-6 attenuates NS5ATP9-induced autophagy slightly. Conclusion: Our results implied that in HCC patients, maybe the higher level of IL-6 in the serum promoted the expression of NS5ATP9 and induced autophagy in cancer cells. And the over-expression of NS5ATP9 which induced by IL-6, in turn, increased IL-6 expression, further, promotes the IL-6/NS5ATP9-mediated autophagy and affects the progression of tumor. Therefore, NS5ATP9 silence might be a potential target for HCC therapy.Keywords: autophagy, Hepatocellular carcinoma, IL-6, microenvironment, NS5ATP9
Procedia PDF Downloads 2517486 Aerobic Exercise Increases Circulating Hematopoietic Stem Cells and Endothelial Progenitor Cells
Authors: Khaled A. shady, Fagr B. Bazeed, Nashwa K. Abousamra, Ihab H. Elberawe, Ashraf E. shaalan, Mohamed A. Sobh
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Physical activity activates a variety of adult stem cells which might be released into the circulation or might be activated in their organ-resident state. A variety of stimuli such as metabolic, mechanical, and hormonal stimuli might by responsible for the mobilization. This study was done to know the changes in hematopoietic stem cells and endothelial progenitor in athletes in the 24 hours following 30 min of aerobic exercise. Methods: Ten healthy male's athlete's (age 20.7± 0.61 y) performed moderate running with 30 min at 80% of velocity of The IAT. Blood samples taken pre-, and immediately, 30 min, 2h, 6h and 24h post-exercise were analyzed for hematopoietic stem cells (HSCs ), endothelial progenitor cells (EPCs(, vascular endothelial growth factor (VEGF), nitric oxide (NO), lactic acid (LA), and white blood cells . HSCs and EPCs were quantified by flow cytometry. Results: After 30min of aerobic exercise significant increases in HSCs, EPC, VEGF, NO, LA and WBCs (p ˂ 0.05). This increase will be at different rates according to the timing of taking blood sample and was in the maximum rate of increase after 30 min of aerobic exercise. HSCs, EPC, NO and WBCs were in the maximum rate of increase 2h post exercise. In addition, VEGF was in the maximum rate of increase immediately post exercise and LA concentration not affected after exercise. Conclusion: These data suggest that HSCs and EPCs increased after aerobic exercise due to increase of VEGF which play an important role in mobilization of stem cells and promotes NO increase which contributes to increase EPCs.Keywords: physical activity, hematopoietic stem cells, mobilization, athletes
Procedia PDF Downloads 1267485 Graphene Materials for Efficient Hybrid Solar Cells: A Spectroscopic Investigation
Authors: Mohammed Khenfouch, Fokotsa V. Molefe, Bakang M. Mothudi
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Nowadays, graphene and its composites are universally known as promising materials. They show their potential in a large field of applications including photovoltaics. This study reports on the role of nanohybrids and nanosystems known as strong light harvesters in the efficiency of graphene hybrid solar cells. Our system included Graphene/ZnO/Porphyrin/P3HT layers. Moreover, the physical properties including surface/interface, optical and vibrational properties were also studied. Our investigations confirmed the interaction between the different components as well as the sensitivity of their photonics to the synthesis conditions. Remarkable energy and charge transfer were detected and deeply investigated. Hence, the optimization of the conditions will lead to the fabrication of higher conversion efficiency in graphene solar cells.Keywords: graphene, optoelectronics, nanohybrids, solar cells
Procedia PDF Downloads 1717484 Induction of Apoptosis by Diosmin through Interleukins/STAT and Mitochondria Mediated Pathway in Hep-2 and KB Cells
Authors: M. Rajasekar, K. Suresh
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Diosmin is a flavonoid, most abundantly found in many citrus fruits. As a flavonoid, it possesses a multitude of biological activities including anti-hyperglycemic, anti-lipid peroxidative, anti-inflammatory, antioxidant, and anti-mutagenic properties. At this point, we established the anti-proliferative and apoptosis-inducing activities of diosmin in Hep-2 and KB cells. Diosmin has cytotoxic effects through inhibiting cellular proliferation of Hep-2 and KB cells, which leads to the induction of apoptosis, as apparent by an increase in the fraction of cells in the sub-G1phase of the cell cycle. Results exposed that inhibition of cell proliferation is associated with regulation of the Interleukins/STAT pathway. In addition, Diosmin treatment with Hep-2 and KB cells actively stimulated reactive oxygen species (ROS) and mitochondrial membrane depolarization. And also an imbalance in the Bax/Bcl-2 ratio triggered the caspase cascade and shifting the balance in favor of apoptosis. These observations conclude that Diosmin induce apoptosis via Interleukins /STAT-mediated pathway.Keywords: diosmin, apoptosis, antioxidant, STAT pathway
Procedia PDF Downloads 3307483 Following the Modulation of Transcriptional Activity of Genes by Chromatin Modifications during the Cell Cycle in Living Cells
Authors: Sharon Yunger, Liat Altman, Yuval Garini, Yaron Shav-Tal
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Understanding the dynamics of transcription in living cells has improved since the development of quantitative fluorescence-based imaging techniques. We established a method for following transcription from a single copy gene in living cells. A gene tagged with MS2 repeats, used for mRNA tagging, in its 3' UTR was integrated into a single genomic locus. The actively transcribing gene was detected and analyzed by fluorescence in situ hybridization (FISH) and live-cell imaging. Several cell clones were created that differed in the promoter regulating the gene. Thus, comparative analysis could be obtained without the risk of different position effects at each integration site. Cells in S/G2 phases could be detected exhibiting two adjacent transcription sites on sister chromatids. A sharp reduction in the transcription levels was observed as cells progressed along the cell cycle. We hypothesized that a change in chromatin structure acts as a general mechanism during the cell cycle leading to down-regulation in the activity of some genes. We addressed this question by treating the cells with chromatin decondensing agents. Quantifying and imaging the treated cells suggests that chromatin structure plays a role both in regulating transcriptional levels along the cell cycle, as well as in limiting an active gene from reaching its maximum transcription potential at any given time. These results contribute to understanding the role of chromatin as a regulator of gene expression.Keywords: cell cycle, living cells, nucleus, transcription
Procedia PDF Downloads 3157482 Induction of G1 Arrest and Apoptosis in Human Cancer Cells by Panaxydol
Authors: Dong-Gyu Leem, Ji-Sun Shin, Sang Yoon Choi, Kyung-Tae Lee
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In this study, we focused on the anti-proliferative effects of panaxydol, a C17 polyacetylenic compound derived from Panax ginseng roots, against various human cancer cells. We treated with panaxydol to various cancer cells and panaxydol treatment was found to significantly inhibit the proliferation of human lung cancer cells (A549) and human pancreatic cancer cells (AsPC-1 and MIA PaCa-2), of which AsPC-1 cells were most sensitive to its treatment. DNA flow cytometric analysis indicated that panaxydol blocked cell cycle progression at the G1 phase in A549 cells, which accompanied by a parallel reduction of protein expression of cyclin-dependent kinase (CDK) 2, CDK4, CDK6, cyclin D1 and cyclin E. CDK inhibitors (CDKIs), such as p21CIP1/WAF1 and p27KIP1, were gradually upregulated after panaxydol treatment at the protein levels. Furthermore, panaxydol induced the activation of p53 in A549 cells. In addition, panaxydol also induced apoptosis of AsPC-1 and MIA PaCa-2 cells, as shown by accumulation of subG1 and apoptotic cell populations. Panaxydol triggered the activation of caspase-3, -8, -9 and the cleavage of poly (ADP-ribose) polymerase (PARP). Reduction of mitochondrial transmembrane potential by panaxydol was determined by staining with dihexyloxacarbocyanine iodide. Furthermore, panaxydol suppressed the levels of anti-apoptotic proteins, XIAP and Bcl-2, and increased the levels of proapoptotic proteins, Bax and Bad. In addition, panaxydol inhibited the activation of Akt and extracellular signal-regulated kinase (ERK) and activated the p38 mitogen-activated protein kinase kinase (MAPK). Our results suggest that panaxydol is an anti-tumor compound that causes p53-mediated cell cycle arrest and apoptosis via mitochondrial apoptotic pathway in various cancer cells.Keywords: apoptosis, cancer, G1 arrest, panaxydol
Procedia PDF Downloads 3247481 Lipid-polymer Nanocarrier Platform Enables X-Ray Induced Photodynamic Therapy against Human Colorectal Cancer Cells
Authors: Rui Sang, Fei Deng, Alexander Engel, Ewa M. Goldys, Wei Deng
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In this study, we brought together X-ray induced photodynamic therapy (X-PDT) and chemo-drug (5-FU) for the treatment on colorectal cancer cells. This was achieved by developing a lipid-polymer hybrid nanoparticle delivery system (FA-LPNPs-VP-5-FU). It was prepared by incorporating a photosensitizer (verteporfin), chemotherapy drug (5-FU), and a targeting moiety (folic acid) into one platform. The average size of these nanoparticles was around 100 nm with low polydispersity. When exposed to clinical doses of 4 Gy X-ray radiation, FA-LPNPs-VP-5-FU generated sufficient amounts of reactive oxygen species, triggering the apoptosis and necrosis pathway of cancer cells. Our combined X-PDT and chemo-drug strategy was effective in inhibiting cancer cells’ growth and proliferation. Cell cycle analyses revealed that our treatment induced G2/M and S phase arrest in HCT116 cells. Our results indicate that this combined treatment provides better antitumour effect in colorectal cancer cells than each of these modalities alone. This may offer a novel approach for effective colorectal cancer treatment with reduced off-target effect and drug toxicity.Keywords: pdt, targeted lipid-polymer nanoparticles, verteporfin, colorectal cancer
Procedia PDF Downloads 817480 Hyaluronic Acid - Alginate Hydrogel for the Transdifferentiation of Testis Cells into Erythrocyte and Hepatocyte-like Cells; A Practice Within an Effective Agent Choice
Authors: Leila Rashki Ghaleno, Mohamad Amin Hajari, Leila Montazeri, Abdolhossein Shahverdi, Mojtaba Rezazadeh Valojerdi
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Background: Spermatogonia stem cells (SSCs) exhibit pluripotency, enabling them to undergo differentiation into many cell lineages, including neurons, glia, endothelial cells, and hepatocytes when cultured in vitro. Although the specific mechanisms are not yet fully understood, it has been observed that biopolymer agents, such as hyaluronic acid (HA) and alginate (Alg), have the potential to induce transdifferentiation of SSCs. The current work aimed to examine the process of in vitro spermatogenesis and the conversion of mouse testicular cells into hepatocytes and erythrocyte-like cells utilizing the HA-Alg hydrogel. Method: After being extracted from the testes of a 5-day postpartum mouse (5 DPP), the testicular cells were separated into two enzymatic stages and then put into a composite hydrogel containing 0.5% HA and 1% alginate. On days 14 and 28 of culture, the colonies' growth, the cells' viability, and their histology were assessed. Result: Despite observing significant cell proliferation on day 14 and the development of circular-shaped organoids on day 28, it was noted that the organoids generated in the HA-Alg medium tended to maintain their circular morphology on day 28. Notably, the testicular cells underwent transdifferentiation into cell types resembling erythrocytes and hepatocytes. The hepatocyte-like cells exhibited the presence of glycogen and lipid deposits, indicating their hepatocyte-like characteristics. Interestingly, immunostaining analysis revealed the secretion of albumin and the presence of VEGFR on day 14. However, on day 28, albumin expression was not detected, while the expression of Sox9 (a marker for hepatocytes), Vegf, CD34, and C-kit (markers for erythrocytes) showed increased levels in the gene expression evaluation. Conclusion: The present findings indicated that HA-Alg could be a potent and effective agent for the transdifferentiation of testis cells into erythrocyte and hepatocyte-like cells, as recent studies have confirmed the transformation of SSCs into hepatocyte cells during in vitro culture.Keywords: 3D culture, mouse testicular cell, hyaluronic acid, liver organoids
Procedia PDF Downloads 727479 Simulation of Remove the Fouling on the in vivo By Using MHD
Authors: Farhad Aalizadeh, Ali Moosavi
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When a blood vessel is injured, the cells of your blood bond together to form a blood clot. The blood clot helps you stop bleeding. Blood clots are made of a combination of blood cells, platelets(small sticky cells that speed up the clot-making process), and fibrin (protein that forms a thread-like mesh to trap cells). Doctors call this kind of blood clot a “thrombus.”We study the effects of different parameters on the deposition of Nanoparticles on the surface of a bump in the blood vessels by the magnetic field. The Maxwell and the flow equations are solved for this purpose. It is assumed that the blood is non-Newtonian and the number of particles has been considered enough to rely on the results statistically. Using MHD and its property it is possible to control the flow velocity, remove the fouling on the walls and return the system to its original form.Keywords: MHD, fouling, in-vivo, blood clots, simulation
Procedia PDF Downloads 4767478 Sitagliptin-AntiCD4 Mab Conjugated T Cell Targeting Therapy for the Effective Treatment of Type I Diabetes
Authors: T. Mahesh, M. K. Samanta
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Antibody dug conjugate (ADC’s) concept is a less explored and more trustable for the treatment of Type 1 diabetes (T1D). T1D is thought to arise from selective immunologically mediated destruction of the insulin- producing β-cells in the pancreatic islets of Langerhans with consequent insulin deficiency. It is evident that type 1 diabetes can be conquered, by 1) to stop immune destruction of βcells, 2) to replace or regenerate β-cells, and 3) to preserve β-cell function and mass. Many studies found that the regulatory T cells (Tregs) are crucial for the maintenance of immunological tolerance. Immune tolerance is liable for the activation of the Th1 response. The important role of Th1 response in pathology of T1D entails the depletion of CD4+ T cells, which initiated the use of anti-CD4 monoclonal antibodies (mAbs) against CD4+ T cells to interfere with induction of T1D.Insulin is regulated by Glucagon-Like Peptide-1 hormone (GLP-1) which also stimulates β-cells proliferation as the half-life of GLP-1 harmone is less due to rapid degradation by DPP-IV enzyme an alternative DPP-IV-inhibitors can increase the half-life of GLP-1 through which it conquers the replacement and reserve β-cells mass. Thus in the present study Anti-CD4 mAb was conjugated with Sitagliptin which is a DPP-IV inhibitor Drug loaded in Nanoparticles through Sulfo-MBS cross-linkers. The above study can be an effective approach for treatment to overcome the Passive subcutaneous insulin therapy.Keywords: antibody drug conjugates, anti-CD4 Mab, DPP IV inhibitors, GLP-1
Procedia PDF Downloads 3927477 Discover a New Technique for Cancer Recognition by Analysis and Determination of Fractal Dimension Images in Matlab Software
Authors: Saeedeh Shahbazkhany
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Cancer is a terrible disease that, if not diagnosed early, therapy can be difficult while it is easily medicable if it is diagnosed in early stages. So it is very important for cancer diagnosis that medical procedures are performed. In this paper we introduce a new method. In this method, we only need pictures of healthy cells and cancer cells. In fact, where we suspect cancer, we take a picture of cells or tissue in that area, and then take some pictures of the surrounding tissues. Then, fractal dimension of images are calculated and compared. Cancer can be easily detected by comparing the fractal dimension of images. In this method, we use Matlab software.Keywords: Matlab software, fractal dimension, cancer, surrounding tissues, cells or tissue, new method
Procedia PDF Downloads 3577476 Treatment of Neuronal Defects by Bone Marrow Stem Cells Differentiation to Neuronal Cells Cultured on Gelatin-PLGA Scaffolds Coated with Nano-Particles
Authors: Alireza Shams, Ali Zamanian, Atefehe Shamosi, Farnaz Ghorbani
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Introduction: Although the application of a new strategy remains a remarkable challenge for treatment of disabilities due to neuronal defects, progress in Nanomedicine and tissue engineering, suggesting the new medical methods. One of the promising strategies for reconstruction and regeneration of nervous tissue is replacing of lost or damaged cells by specific scaffolds after Compressive, ischemic and traumatic injuries of central nervous system. Furthermore, ultrastructure, composition, and arrangement of tissue scaffolds are effective on cell grafts. We followed implantation and differentiation of mesenchyme stem cells to neural cells on Gelatin Polylactic-co-glycolic acid (PLGA) scaffolds coated with iron nanoparticles. The aim of this study was to evaluate the capability of stem cells to differentiate into motor neuron-like cells under topographical cues and morphogenic factors. Methods and Materials: Bone marrow mesenchymal stem cells (BMMSCs) was obtained by primary cell culturing of adult rat bone marrow got from femur bone by flushing method. BMMSCs were incubated with DMEM/F12 (Gibco), 15% FBS and 100 U/ml pen/strep as media. Then, BMMSCs seeded on Gel/PLGA scaffolds and tissue culture (TCP) polystyrene embedded and incorporated by Fe Nano particles (FeNPs) (Fe3o4 oxide (M w= 270.30 gr/mol.). For neuronal differentiation, 2×10 5 BMMSCs were seeded on Gel/PLGA/FeNPs scaffolds was cultured for 7 days and 0.5 µ mol. Retinoic acid, 100 µ mol. Ascorbic acid,10 ng/ml. Basic fibroblast growth factor (Sigma, USA), 250 μM Iso butyl methyl xanthine, 100 μM 2-mercaptoethanol, and 0.2 % B27 (Invitrogen, USA) added to media. Proliferation of BMMSCs was assessed by using MTT assay for cell survival. The morphology of BMMSCs and scaffolds was investigated by scanning electron microscopy analysis. Expression of neuron-specific markers was studied by immunohistochemistry method. Data were analyzed by analysis of variance, and statistical significance was determined by Turkey’s test. Results: Our results revealed that differentiation and survival of BMMSCs into motor neuron-like cells on Gel/PLGA/FeNPs as a biocompatible and biodegradable scaffolds were better than those cultured in Gel/PLGA in absence of FeNPs and TCP scaffolds. FeNPs had raised physical power but decreased capacity absorption of scaffolds. Well defined oriented pores in scaffolds due to FeNPs may activate differentiation and synchronized cells as a mechanoreceptor. Induction effects of magnetic FeNPs by One way flow of channels in scaffolds help to lead the cells and can facilitate direction of their growth processes. Discussion: Progression of biological properties of BMMSCs and the effects of FeNPs spreading under magnetic field was evaluated in this investigation. In vitro study showed that the Gel/PLGA/FeNPs scaffold provided a suitable structure for motor neuron-like cells differentiation. This could be a promising candidate for enhancing repair and regeneration in neural defects. Dynamic and static magnetic field for inducing and construction of cells can provide better results for further experimental studies.Keywords: differentiation, mesenchymal stem cells, nano particles, neuronal defects, Scaffolds
Procedia PDF Downloads 1697475 Genotoxic and Cytotoxic Effects of Methidathion Pesticide
Authors: Mohammad Y. Alfaifi
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Methidathion (MTD) (Trade name Supracide®) is a non-systemic organophosphorus insecticide used intensively worldwide including Saudi Arabia. However, there is a lack in published studies about it's genotoxicity. In this study we evaluated MTD toxicity in rat bone marrow cells (in vivo) and in lymphocytes (in vitro) using different doses based on LD50. MNNCE (Micronucleated normocromatic erythrocytes) and MNPCE (Micronucleated polychromatic erythrocytes), NDI (Nuclear division index) and NDCI (nuclear division cytotoxicity index), necrotic and apoptotic cells were recorded in rat's bone marrow samples. CA, MI (number of cells undergoing mitosis) necrotic, and apoptotic cells recorded in lymphocytes. Results showed that there was a slight increase in the frequency of micronucleated bone marrow cells. However, no structural chromosomal aberrations were detected in vivo or in vitro. On the other hand, the results showed significant increase in necrotic and apoptotic cells following MTD administration in a dose-dependent manner comparing to positive and negative control groups. In light of these results, MTD can be considered highly cytotoxic and moderate genotoxic, and precaution should be taken when using MTD.Keywords: methidathion, micronucleus, NDI, NDCI, toxicity, chromosomal aberrations
Procedia PDF Downloads 418