Search results for: induced pluripotent stem cells
5655 Nitrate-Induced Biochemical and Histopathological Changes in the Kidney of Rats: Attenuation by Hyparrhenia hirta
Authors: Hanen Bouaziz, Moez Rafrafi, Ghada Ben Salah, Kamel Jamoussi, Tahia Boudawara, Najiba Zeghal
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The present study investigated the protective role of Hyparrhenia hirta against sodium nitrate (NaNO3)-induced nephrotoxicity. A high-performance liquid chromatography coupled with a mass spectrometer (HPLC-MS) method was developed to separate and identify flavonoids in Hyparrhenia hirta. Seven flavonoids were identified as 3-O-methylquercetin, luteolin-7-O-glucoside, luteolin, apigenin-7-O-glucoside, apigenin-8-C-glucoside, luteolin-8-C-glucoside and luteolin-6-C-glucoside. Wistar rats were randomly divided into three groups: a control group and two treated groups during 50 days with NaNO3 administered either alone in drinking water or co-administered with Hyparrhenia hirta. NaNO3 treatment induced a significant increase in plasma levels of creatinine, urea and uric while urinary level decreased significantly. Nephrotoxicity induced by NaNO3 was characterized by significant increase in creatinine clearance. In parallel, a significant increase in malondialdehyde level along with a concomitant decrease in total glutathione content and superoxide dismutase, catalase and glutathione peroxidase activities were observed in the kidney after NaNO3 treatment. The histopathological changes in kidney after NaNO3 administration were shrunken. There were renal tubule cell degeneration and infiltration of mononuclear cells. Most glomeruli revealed shrinkage, a wide capsular space and a peri-glomerular mononuclear cells infiltration. Hyparrhenia hirta supplementation showed a remarkable amelioration of the abnormalities cited above. The results concluded that the treatment with Hyparrhenia hirta had a significant role in protecting the animals from nitrate-induced kidney dysfunction.Keywords: flavonoids, hyparrhenia hirta, kidney, nitrate toxicity, oxidative stress, rat
Procedia PDF Downloads 4455654 The Antidiabetic Properties of Indonesian Swietenia mahagoni in Alloxan-Induced Diabetic Rats
Authors: T. Wresdiyati, S. Sa’diah, A. Winarto
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Diabetes mellitus (DM) is a metabolic disease that can be indicated by the high level of blood glucose. The objective of this study was to observe the antidiabetic properties of ethanolic extract of Indonesian Swietenia mahagoni Jacq. seed on the profile of pancreatic superoxide dismutase and β-cells in the alloxan- experimental diabetic rats. The Swietenia mahagoni seed was obtained from Leuwiliang-Bogor, Indonesia. Extraction of Swietenia mahagoni was done by using ethanol with maceration methods. A total of 25 male Sprague dawley rats were divided into five groups; (a) negative control group, (b) positive control group (DM), (c) DM group that was treated with Swietenia mahagoni seed extract, (d) DM group that was treated with acarbose, and (e) non-DM group that was treated with Swietenia mahagoni seed extract. The DM groups were induced by alloxan (110 mg/kgBW). The extract was orally administrated to diabetic rats 500 mg/kg/BW/day for 28 days. The extract showed hypoglycemic effect, increased body weight, increased the content of superoxide dismutase in the pancreatic tissue, and delayed the rate of β-cells damage of experimental diabetic rats. These results suggested that the ethanolic extract of Indonesian Swietenia mahagoni Jacq. seed could be proposed as a potential anti-diabetic agent.Keywords: beta cells, diabetes, hypoglycemic, rat, Swietenia mahagoni
Procedia PDF Downloads 2955653 Treatment of Full-Thickness Rotator Cuff Tendon Tear Using Umbilical Cord Blood-Derived Mesenchymal Stem Cells and Polydeoxyribonucleotides in a Rabbit Model
Authors: Sang Chul Lee, Gi-Young Park, Dong Rak Kwon
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Objective: The aim of this study was to investigate regenerative effects of ultrasound (US)-guided injection with human umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) and/or polydeoxyribonucleotide (PDRN) injection in a chronic traumatic full-thickness rotator cuff tendon tear (FTRCTT) in a rabbit model. Material and Methods: Rabbits (n = 32) were allocated into 4 groups. After a 5-mm sized FTRCTT just proximal to the insertion site on the subscapularis tendon was created by excision, the wound was immediately covered by silicone tube to prevent natural healing. After 6 weeks, 4 injections (0.2 mL normal saline, G1; 0.2 mL PDRN, G2; 0.2 mL UCB-MSCs, G3; and 0.2 mL UCB-MSCs with 0.2ml PDRN, G4) were injected into FTRCTT under US guidance. We evaluated gross morphologic changes on all rabbits after sacrifice. Masson’s trichrome, anti-type 1 collagen antibody, bromodeoxyuridine, proliferating cell nuclear antigen, vascular endothelial growth factor and platelet endothelial cell adhesion molecule stain were performed to evaluate histological changes. Motion analysis was also performed. Results: The gross morphologic mean tendon tear size in G3 and 4 was significantly smaller than that of G1 and 2 (p < .05). However, there were no significant differences in tendon tear size between G3 and 4. In G4, newly regenerated collagen type 1 fibers, proliferating cells activity, angiogenesis, walking distance, fast walking time, and mean walking speed were greater than in the other three groups on histological examination and motion analysis. Conclusion: Co-injection of UCB-MSCs and PDRN was more effective than UCB-MSCs injection alone in histological and motion analysis in a rabbit model of chronic traumatic FTRCTT. However, there was no significant difference in gross morphologic change of tendon tear between UCB-MSCs with/without PDRN injection. The results of this study regarding the combination of UCB-MSCs and PDRN are worth additional investigations.Keywords: mesenchymal stem cell, umbilical cord, polydeoxyribonucleotides, shoulder, rotator cuff, ultrasonography, injections
Procedia PDF Downloads 1855652 Integrative Biology Teaching and Learning Model Based on STEM Education
Authors: Narupot Putwattana
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Changes in global situation such as environmental and economic crisis brought the new perspective for science education called integrative biology. STEM has been increasingly mentioned for several educational researches as the approach which combines the concept in Science (S), Technology (T), Engineering (E) and Mathematics (M) to apply in teaching and learning process so as to strengthen the 21st-century skills such as creativity and critical thinking. Recent studies demonstrated STEM as the pedagogy which described the engineering process along with the science classroom activities. So far, pedagogical contents for STEM explaining the content in biology have been scarce. A qualitative literature review was conducted so as to gather the articles based on electronic databases (google scholar). STEM education, engineering design, teaching and learning of biology were used as main keywords to find out researches involving with the application of STEM in biology teaching and learning process. All articles were analyzed to obtain appropriate teaching and learning model that unify the core concept of biology. The synthesized model comprised of engineering design, inquiry-based learning, biological prototype and biologically-inspired design (BID). STEM content and context integration were used as the theoretical framework to create the integrative biology instructional model for STEM education. Several disciplines contents such as biology, engineering, and technology were regarded for inquiry-based learning to build biological prototype. Direct and indirect integrations were used to provide the knowledge into the biology related STEM strategy. Meanwhile, engineering design and BID showed the occupational context for engineer and biologist. Technological and mathematical aspects were required to be inspected in terms of co-teaching method. Lastly, other variables such as critical thinking and problem-solving skills should be more considered in the further researches.Keywords: biomimicry, engineering approach, STEM education, teaching and learning model
Procedia PDF Downloads 2575651 Anti-Cancerous Activity of Sargassum siliquastrum in Cervical Cancer: Choreographing the Fly's Danse Macabre
Authors: Sana Abbasa, Shahzad Bhattiab, Nadir Khan
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Sargassum siliquastrum is brown seaweed with traditional claims for some medicinal properties. This research was done to investigate the methanol extract of S. siliquastrum for antiproliferative activity against human cervical cancer cell line, HeLa and its mode of cell death. From methylene blue assay, S. siliquastrum exhibited antiproliferative activity on HeLa cells with IC50 of 3.87 µg/ml without affecting non-malignant cells. Phase contrast microscopy indicated the confluency reduction in HeLa cells and changes on the cell shape. Nuclear staining with Hoechst 33258 displayed the formation of apoptotic bodies and fragmented nuclei. S. siliquastrum also induced early apoptosis event in HeLa cells as confirmed by FITC-Annexin V/propidium iodide staining by flow cytometry analysis. Cell cycle analysis indicated growth arrest of HeLa cells at G1/S phase. Protein study by flow cytometry indicated the increment of p53, slight increase of Bax and unchanged level of Bcl-2. In conclusion, S. siliquastrum demonstrated an antiproliferative activity in HeLa cell by inducing G1/S cell cycle arrest via p53-mediated pathway.Keywords: sargassum siliquastrum, cervical cancer, P53, antiproleferation
Procedia PDF Downloads 6335650 Copper Chelation by 3-(Bromoacetyl) Coumarin Derivative Induced Apoptosis in Cancer Cells: Influence of Copper Chelation Strategy in Cancer Treatment
Authors: Saman Khan, Imrana Naseem
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Copper is an essential trace element required for pro-angiogenic co-factors including vascular endothelial growth factor (VEGF). Elevated levels of copper are found in various types of cancer including prostrate, colon, breast, lung and liver for angiogensis and metastasis. Therefore, targeting copper via copper-specific chelators in cancer cells can be developed as effective anticancer treatment strategy. In continuation of our pursuit to design and synthesize copper chelators, herein we opted for a reaction to incorporate di-(2-picolyl) amine in 3-(bromoacetyl) coumarin (parent backbone) for the synthesis of complex 1. We evaluated lipid peroxidation, protein carbonylation, ROS generation, DNA damage and consequent apoptosis by complex 1 in exogenously added Cu(II) in human peripheral lymphocytes (simulate malignancy condition). Results showed that Cu(II)-complex 1 interaction leads to cell proliferation inhibition, apoptosis, ROS generation and DNA damage in human lymphocytes, and these effects were abrogated by cuprous chelator neocuproine and ROS scavengers (thiourea, catalase, SOD). This indicates that complex 1 cytotoxicity is due to redox cycling of copper to generate ROS which leads to pro-oxidant cell death in cancer cells. To further confirm our hypothesis, using the rat model of diethylnitrosamine (DEN) induced hepatocellular carcinoma; we showed that complex 1 mediates DNA breakage and cell death in isolated carcinoma cells. Membrane permeant copper chelator, neocuproine, and ROS scavengers inhibited the complex 1-mediated cellular DNA degradation and apoptosis. In summary, complex 1 anticancer activity is due to its copper chelation capability. These results will provide copper chelation as an effective targeted cancer treatment strategy for selective cytotoxic action against malignant cells without affecting normal cells.Keywords: cancer treatment, copper chelation, ROS generation, DNA damage, redox cycling, apoptosis
Procedia PDF Downloads 2925649 Machine Learning and Metaheuristic Algorithms in Short Femoral Stem Custom Design to Reduce Stress Shielding
Authors: Isabel Moscol, Carlos J. Díaz, Ciro Rodríguez
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Hip replacement becomes necessary when a person suffers severe pain or considerable functional limitations and the best option to enhance their quality of life is through the replacement of the damaged joint. One of the main components in femoral prostheses is the stem which distributes the loads from the joint to the proximal femur. To preserve more bone stock and avoid weakening of the diaphysis, a short starting stem was selected, generated from the intramedullary morphology of the patient's femur. It ensures the implantability of the design and leads to geometric delimitation for personalized optimization with machine learning (ML) and metaheuristic algorithms. The present study attempts to design a cementless short stem to make the strain deviation before and after implantation close to zero, promoting its fixation and durability. Regression models developed to estimate the percentage change of maximum principal stresses were used as objective optimization functions by the metaheuristic algorithm. The latter evaluated different geometries of the short stem with the modification of certain parameters in oblique sections from the osteotomy plane. The optimized geometry reached a global stress shielding (SS) of 18.37% with a determination factor (R²) of 0.667. The predicted results favour implantability integration in the short stem optimization to effectively reduce SS in the proximal femur.Keywords: machine learning techniques, metaheuristic algorithms, short-stem design, stress shielding, hip replacement
Procedia PDF Downloads 1965648 Impact of Light Intensity, Illumation Strategy and Self-Shading on Sustainable Algal Growth in Photo Bioreactors
Authors: Amritanshu Shriwastav, Purnendu Bose
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Algal photo bioreactors were operated at incident light intensities of 0.24, 2.52 and 5.96 W L-1 to determine the impact of light on algal growth. Low specific Chlorophyll-a content of algae was a strong indicator of light induced stress on algal cells. It was concluded that long term operation of photo bioreactors in the continuous illumination mode was infeasible under the range of incident light intensities examined and provision of a dark period after each light period was necessary for algal cells to recover from light-induced stress. Long term operation of photo bioreactors in the intermittent illumination mode was however possible at light intensities of 0.24 and 2.52 W L-1. Further, the incident light intensity in the photo bioreactors was found to decline exponentially with increase in algal concentration in the reactor due to algal ‘self-shading’. This may be an important determinant for photo bioreactor performance at higher algal concentrations.Keywords: Algae, algal growth, photo bioreactor, photo-inhibition, ‘self-shading’
Procedia PDF Downloads 3215647 Reconstruction of Alveolar Bone Defects Using Bone Morphogenetic Protein 2 Mediated Rabbit Dental Pulp Stem Cells Seeded on Nano-Hydroxyapatite/Collagen/Poly(L-Lactide)
Authors: Ling-Ling E., Hong-Chen Liu, Dong-Sheng Wang, Fang Su, Xia Wu, Zhan-Ping Shi, Yan Lv, Jia-Zhu Wang
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Objective: The objective of the present study is to evaluate the capacity of a tissue-engineered bone complex of recombinant human bone morphogenetic protein 2 (rhBMP-2) mediated dental pulp stem cells (DPSCs) and nano-hydroxyapatite/collagen/poly(L-lactide)(nHAC/PLA) to reconstruct critical-size alveolar bone defects in New Zealand rabbit. Methods: Autologous DPSCs were isolated from rabbit dental pulp tissue and expanded ex vivo to enrich DPSCs numbers, and then their attachment and differentiation capability were evaluated when cultured on the culture plate or nHAC/PLA. The alveolar bone defects were treated with nHAC/PLA, nHAC/PLA+rhBMP-2, nHAC/PLA+DPSCs, nHAC/PLA+DPSCs+rhBMP-2, and autogenous bone (AB) obtained from iliac bone or were left untreated as a control. X-ray and a polychrome sequential fluorescent labeling were performed post-operatively and the animals were sacrificed 12 weeks after operation for histological observation and histomorphometric analysis. Results: Our results showed that DPSCs expressed STRO-1 and vementin, and favoured osteogenesis and adipogenesis in conditioned media. DPSCs attached and spread well, and retained their osteogenic phenotypes on nHAC/PLA. The rhBMP-2 could significantly increase protein content, alkaline phosphatase (ALP) activity/protein, osteocalcin (OCN) content, and mineral formation of DPSCs cultured on nHAC/PLA. The X-ray graph, the fluorescent, histological observation and histomorphometric analysis showed that the nHAC/PLA+DPSCs+rhBMP-2 tissue-engineered bone complex had an earlier mineralization and more bone formation inside the scaffold than nHAC/PLA, nHAC/PLA+rhBMP-2 and nHAC/PLA+DPSCs, or even autologous bone. Implanted DPSCs contribution to new bone were detected through transfected eGFP genes. Conclutions: Our findings indicated that stem cells existed in adult rabbit dental pulp tissue. The rhBMP-2 promoted osteogenic capability of DPSCs as a potential cell source for periodontal bone regeneration. The nHAC/PLA could serve as a good scaffold for autologous DPSCs seeding, proliferation and differentiation. The tissue-engineered bone complex with nHAC/PLA, rhBMP-2, and autologous DPSCs might be a better alternative to autologous bone for the clinical reconstruction of periodontal bone defects.Keywords: nano-hydroxyapatite/collagen/poly (L-lactide), dental pulp stem cell, recombinant human bone morphogenetic protein, bone tissue engineering, alveolar bone
Procedia PDF Downloads 4025646 The Role Of Diallyl Trisulfide As A Suppressor In Activated-Platelets Induced Human Breast Cancer MDA-MB-435s Cells Hematogenous Metastasis
Authors: Yuping Liu, Li Tao, Yin Lu
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Accumulating evidence has been shown that diallyl trisulfide (DATS) from garlic may reduce the risk of developing several types of cancer. In view of the dynamic crosstalk interplayed by tumor cells and platelets in hematogenous metastasis, we demonstrate the effectiveness of DATS on the metastatic behaviors of MDA-MB-435s human breast cancer cell line co-incubated with activated platelets. Indeed, our data identified that DATS significantly blocked platelets fouction induced by PAF, followed by the decreased production of TXB2. DATS was found to dose-dependently suppressed MDA-MB-435s cell migration and invasion in presence of activated platelets by PAF in vitro. Furthermore, the expression, secretion and enzymatic activity of matrix metalloproteinase (MMP)-2/9, as well as the luciferase activity of upstream regulator NF-κB in MDA-MB-435s, were obviously diminished by DATS. In parallel, DATS blocked upstream NF-κB activation signaling complexes composed of extracellular signal-related kinase (ERK) as assessed by measuring the levels of the phosphorylated forms.Keywords: DATS, ERK, metastasis, MMPs, NF-κB, platelet
Procedia PDF Downloads 3875645 Radix Saposhnikoviae Suppresses Allergic Contact Dermatitis in Mice by Regulating DCs Activated Th1-Type Cells
Authors: Hailiang Liu, Yan Ni, Jie Zheng, Xiaoyan Jiang, Min Hong
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Allergic contact dermatitis (ACD) is a commonly clinical type IV allergic skin disease, with the pathological features of infiltration by mononuclear cells and tissue necrosis. Traditional Chinese medicine Radix Saposhnikoviae (RS) is traditionally employed to treat exogenous evils, rubella, itching, rheumatism and tetanus. Meanwhile, it is an important component of the commonly used anti-allergy compound. It’s now widely used as an immuno-modulating agent in mixed herbal decoctions to treat inflammation. However, its mechanism of anti-allergy remains unknown. RS was found to reduce ear thickness, as well as the infiltration of eosinophils. The proliferation of T lymphocytes was inhibited significantly by RS, markedly decreased IFN-γ levels in the supernatant of cells cultured and serum were detected with the treatment of RS. RS significantly decreased the amount of DCs in the mouse lymph nodes, and inhibited the expression of CD4 0 and CD86. Meanwhile, T-bet mRNA expression was down remarkably regulated by RS. These results indicate that RS cures Th1-induced allergic skin inflammation by regulating Th1/Th2 balance with decreasing Th1 differentiation, which might be associated with DCs.Keywords: allergic contact dermatitis, Radix saposhnikoviae, dendritic cells, T-bet, GATA-3, CD4+ CD25+ Foxp3+ treg cells
Procedia PDF Downloads 3745644 Intracellular Sphingosine-1-Phosphate Receptor 3 Contributes to Lung Tumor Cell Proliferation
Authors: Michela Terlizzi, Chiara Colarusso, Aldo Pinto, Rosalinda Sorrentino
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Sphingosine-1-phosphate (S1P) is a membrane-derived bioactive phospholipid exerting a multitude of effects on respiratory cell physiology and pathology through five S1P receptors (S1PR1-5). Higher levels of S1P have been registered in a broad range of respiratory diseases, including inflammatory disorders and cancer, although its exact role is still elusive. Based on our previous study in which we found that S1P/S1PR3 is involved in an inflammatory pattern via the activation of Toll-like Receptor 9 (TLR9), highly expressed on lung cancer cells, the main goal of the current study was to better understand the involvement of S1P/S1PR3 pathway/signaling during lung carcinogenesis, taking advantage of a mouse model of first-hand smoke exposure and of carcinogen-induced lung cancer. We used human samples of Non-Small Cell Lung Cancer (NSCLC), a mouse model of first-hand smoking, and of Benzo(a)pyrene (BaP)-induced tumor-bearing mice and A549 lung adenocarcinoma cells. We found that the intranuclear, but not the membrane, localization of S1PR3 was associated to the proliferation of lung adenocarcinoma cells, the mechanism that was correlated to human and mouse samples of smoke-exposure and carcinogen-induced lung cancer, which were characterized by higher utilization of S1P. Indeed, the inhibition of the membrane S1PR3 did not alter tumor cell proliferation after TLR9 activation. Instead, according to the nuclear localization of sphingosine kinase (SPHK) II, the enzyme responsible for the catalysis of the S1P last step synthesis, the inhibition of the kinase completely blocked the endogenous S1P-induced tumor cell proliferation. These results prove that the endogenous TLR9-induced S1P can on one side favor pro-inflammatory mechanisms in the tumor microenvironment via the activation of cell surface receptors, but on the other tumor progression via the nuclear S1PR3/SPHK II axis, highlighting a novel molecular mechanism that identifies S1P as one of the crucial mediators for lung carcinogenesis-associated inflammatory processes and that could provide differential therapeutic approaches especially in non-responsive lung cancer patients.Keywords: sphingosine-1-phosphate (S1P), S1P Receptor 3 (S1PR3), smoking-mice, lung inflammation, lung cancer
Procedia PDF Downloads 2015643 The Balancing of the Parental Responsibilities and Right and the Best Interest of the Child within the Parent-Child Relationship
Authors: R. Prinsloo
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Amniotic fluid stem cells (AFSC) have been shown to contribute towards the amelioration of Acute Renal Failure (ARF), but the mechanisms underlying the renoprotective effect are largely unknown. Therefore, the main goal of the current study was to evaluate the therapeutic efficacy of AFSC in a cisplatin-induced rat model of ARF and to investigate the underlying mechanisms responsible for its renoprotective effect. To study the therapeutic efficacy of AFSC, ARF was induced in Wistar rats by an intra-peritoneal injection of cisplatin, and five days after administration, the rats were randomized into two groups and injected with either AFSC or normal saline intravenously. On day 8 and 12 after cisplatin injection, i.e., day 3 and day7 post-therapy respectively, the blood biochemical parameters, histopathological changes, apoptosis, and expression of pro-apoptotic, anti-apoptotic and autophagy-related proteins in renal tissues were studied in both groups of rats. Administration of AFSC in ARF rats resulted in improvement of renal function and attenuation of renal damage as reflected by significant decrease in blood urea nitrogen, serum creatinine levels, tubular cell apoptosis as assessed by Bax/Bcl2 ratio, and expression of the pro-apoptotic proteins viz. PUMA, Bax, cleaved caspase-3 and cleaved caspase-9 as compared to saline-treated group. Furthermore, in the AFSC-treated group as compared to saline-treated group, there was a significant increase in the activation of autophagy as evident by increased expression of LC3-II, ATG5, ATG7, Beclin1 and phospho-AMPK levels with a concomitant decrease in phospho-p70S6K and p62 expression levels. To further confirm whether the protective effects of AFSC on cisplatin-induced apoptosis were dependent on autophagy, chloroquine, an autophagy inhibitor was administered by the intra-peritoneal route. Chloroquine administration led to significant reduction in the anti-apoptotic effects of the AFSC therapy and further deterioration in the renal structure and function caused by cisplatin. Collectively, our results put forth that AFSC ameliorates cisplatin-induced ARF through induction of autophagy and inhibition of apoptosis. Furthermore, the protective effects of AFSC were blunted by chloroquine, highlighting that activation of autophagy is an important mechanism of action for the protective role of AFSC in cisplatin-induced renal injury.Keywords: best interest of the child, children's rights, parent and child relationship, parental responsibilities and rights
Procedia PDF Downloads 1055642 Hydrogel Based on Cellulose Acetate Used as Scaffold for Cell Growth
Authors: A. Maria G. Melero, A. M. Senna, J. A. Domingues, M. A. Hausen, E. Aparecida R. Duek, V. R. Botaro
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A hydrogel from cellulose acetate cross linked with ethylenediaminetetraacetic dianhydride (HAC-EDTA) was synthesized by our research group, and submitted to characterization and biological tests. Cytocompatibility analysis was performed by confocal microscopy using human adipocyte derived stem cells (ASCs). The FTIR analysis showed characteristic bands of cellulose acetate and hydroxyl groups and the tensile tests evidence that HAC-EDTA present a Young’s modulus of 643.7 MPa. The confocal analysis revealed that there was cell growth at the surface of HAC-EDTA. After one day of culture the cells presented spherical morphology, which may be caused by stress of the sequestration of Ca2+ and Mg2+ ions at the cell medium by HAC-EDTA, as demonstrated by ICP-MS. However, after seven days and 14 days of culture, the cells present fibroblastoid morphology, phenotype expected by this cellular type. The results give efforts to indicate this new material as a potential biomaterial for tissue engineering, in the future in vivo approach.Keywords: cellulose acetate, hydrogel, biomaterial, cellular growth
Procedia PDF Downloads 1955641 Chemotactic Behaviour of Human Mesenchymal Stem Cells in Response to Silicate Substituted Hydroxyapatite
Authors: Dinara Ikramova, Karin A. Hing, Simon C. F. Rawlinson
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Silicate-substituted hydroxyapatite (SiHA) has been shown to enhance bone regeneration in vivo compared with phase pure stoichiometric hydroxyapatite. Evidence suggests that substrate chemistry dependent formation of a permissive protein layer on the surface of synthetic bone graft substitute materials is key for bioactivity and cell attachment. However, little information is available on whether the substrate chemistry may affect cell migration and recruitment. The aim of this study is to investigate whether or not human Mesenchymal Stem Cells (hMSCs) exhibit a chemotactic response to SiHA porous granules and if it can be linked to either the ion exchange or protein sequestering and enrichment on the surface of the material. 150mg of SiHA granules with 80% total porosity and 20% strut porosity were incubated in 1ml of either Serum Free Media (SFM) or 10% Serum Containing Media (SCM) under static cell culture conditions (37°C, 5% CO2) in absence of cells. Protein sequestering and exchange of calcium, phosphate and silicate ions were analysed at 0.5, 1, 2, 4, 8, 16 and 24 hours with n=12 per time point. Migration of hMSCs in the presence of 150mg of SiHA granules was assessed over 24 hours using a modified transwell migration system in either SFM or SCM (n=6) with 30% serum containing media acting as a positive control. At 24 hours protein sequestering and ionic exchange were analysed, and the number of cells was quantified using a high throughput confocal microscope (IN Cell Analyser 6000). In acellular condition, both calcium and phosphate ion concentrations in media showed a decrease at 24 hours which was greater in SFM than in SCM. This suggests possible formation and precipitation of a bone like apatite on the surface of SiHA. Reduction in this activity observed in SCM indicates that the presence of serum proteins is interfering with the ion exchange at the material and media interface. Adsorbed protein levels showed fluctuation over time followed by sharp decrease at 24 hours, suggesting a possible protein rearrangement on the surface of the material. The ion analysis performed on SFM and SCM after 24-hour incubation with cells in the presence of granules showed a greater reduction in phosphate concentration in both SFM and SCM compared to phosphate levels in acellular condition. Silicate concentration in SCM increased from 1.6mM (absence of cells) to 5.1mM (presence of cells). This indicates that the cells are promoting the uptake of phosphate and release of silicate ions. No significant change was seen in levels of adsorbed proteins in the presence and absence of cells. Further analysis is required to determine whether the species of these proteins change over time. The analysis of cell migration after 24-hour incubation showed more cells migrating towards the granules, 12.7% in SFM and 8.3% in SCM, than in positive control, 4.5% in SFM and 3.6% in SCM respectively. These results suggest that SiHA has a chemotactic activity independent of serum proteins. A property which has not previously been demonstrated for a synthetic bone graft material.Keywords: cell migration, hMSCs, SiHA, transwell migration system
Procedia PDF Downloads 1325640 Establishment of a Thermostable Newcastle Disease Vaccine Candidate Strain and Its Adaptation to Vero Cells
Authors: Humayun Kabir, Amirul Hasan, Yu Miyaoka, Makiko Yamaguchi, Chisaki Kadota, Kazuaki Takehara
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From field isolates of Newcastle disease virus (NDV) in Japan, one avirulent strain, APMV/northern pintail/Japan/Aomori/2003 (dk-Aomori/03, NDV 261), was selected for its excellent thermostability, and the strain was heat-treated at 56℃ temperatures for 30 min with each passage into Vero cells to maintain thermostability and to adapt Vero cells. After serial 20 passages in Vero cells, it was named NDV Vero20. When growth curves were tested in Vero cells, NDV Vero20 grew well to compare the original NDV261. The HN gene was sequenced, and found motifs that show thermostability. The intracerebral pathogenicity index (ICPI) test score was 0. The thermostability of the virus was confirmed by storing it at different temperatures, including at 37°C. When susceptible chicks were inoculated with NDV Vero20 through eye drops, induced adequate levels of antibody were measured using a serum neutralization test. The results showed that NDV Vero20, a vaccine candidate strain is thermostable, Vero cell adapted, and has immunogenic potential, which would make as an alternative to the traditional embryonated chicken eggs-based vaccine.Keywords: Newcastle disease virus, thermostability, vaccine, Vero cell adaptability
Procedia PDF Downloads 1425639 Non-Canonical Beclin-1-Independent Autophagy and Apoptosis in Cell Death Induced by Rhus coriaria in Human Colon HT-29 Cancer Cells
Authors: Rabah Iratni, Husain El Hasasna, Khawlah Athamneh, Halima Al Sameri, Nehla Benhalilou, Asma Al Rashedi
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Background: Cancer therapies have witnessed great advances in the recent past, however, cancer continues to be a leading cause of death, with colorectal cancer being the fourth cause of cancer-related deaths. Colorectal cancer affects both sexes equally with poor survival rate once it metastasizes. Phytochemicals, which are plant derived compounds, have been on a steady rise as anti-cancer drugs due to the accumulation of evidences that support their potential. Here, we investigated the anticancer effect of Rhus coriaria on colon cancer cells. Material and Method: Human colon cancer HT-29 cell line was used. Protein expression and protein phosphorylation were examined using Western blotting. Transcription activity was measure using Quantitative RT-PCR. Human tumoral clonogenic assay was used to assess cell survival. Senescence was assessed by the senescence-associated beta-galactosidase assay. Results: Rhus coriaria extract (RCE) was found to significantly inhibit the viability and colony growth of human HT-29 colon cancer cells. RCE induced senescence and cell cycle arrest at G1 phase. These changes were concomitant with upregulation of p21, p16, downregulation of cyclin D1, p27, c-myc and expression of Senescence-associated-β-Galactosidase activity. Moreover, RCE induced non-canonical beclin-1independent autophagy and subsequent apoptotic cell death through activation of activation caspase 8 and caspase 7. The blocking of autophagy by 3-methyladenine (3-MA) or chloroquine (CQ) reduced RCE-induced cell death. Further, RCE induced DNA damage, reduced mutant p53 protein level and downregulated phospho-AKT and phospho-mTOR, events that preceded autophagy. Mechanistically, we found that RCE inhibited the AKT and mTOR pathway, a regulator of autophagy, by promoting the proteasome-dependent degradation of both AKT and mTOR proteins. Conclusion: Our findings provide strong evidence that Rhus coriaria possesses strong anti-colon cancer activity through induction of senescence and autophagic cell death, making it a promising alternative or adjunct therapeutic candidate against colon cancer.Keywords: autophagy, proteasome degradation, senescence, mTOR, apoptosis, Beclin-1
Procedia PDF Downloads 2635638 Comparison of the Performance of GaInAsSb and GaSb Cells under Different Temperature Blackbody Radiations
Authors: Liangliang Tang, Chang Xu, Xingying Chen
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GaInAsSb cells probably show better performance than GaSb cells in low-temperature thermophotovoltaic systems due to lower bandgap; however, few experiments proved this phenomenon so far. In this paper, numerical simulation is used to evaluate GaInAsSb and GaSb cells with similar structures under different radiation temperatures. We found that GaInAsSb cells with n-type emitters show slightly higher output power densities compared with that of GaSb cells with n-type emitters below 1,550 K-blackbody radiation, and the power density of the later cells will suppress the formers above this temperature point. During the temperature range of 1,000~2,000 K, the efficiencies of GaSb cells are about twice of GaInAsSb cells if perfect filters are used to prevent the emission of the non-absorbed long wavelength photons. Several parameters that affect the GaInAsSb cell were analyzed, such as doping profiles, thicknesses of GaInAsSb epitaxial layer and surface recombination velocity. The non-p junctions, i.e., n-type emitters are better for GaInAsSb cell fabrication, which is similar to that of GaSb cells.Keywords: thermophotovoltaic cell, GaSb, GaInAsSb, diffused emitters
Procedia PDF Downloads 2815637 Integration of STEM Education in Quebec, Canada – Challenges and Opportunities
Authors: B. El Fadil, R. Najar
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STEM education is promoted by many scholars and curricula around the world, but it is not yet well established in the province of Quebec in Canada. In addition, effective instructional STEM activities and design methods are required to ensure that students and teachers' needs are being met. One potential method is the Engineering Design Process (EDP), a methodology that emphasizes the importance of creativity and collaboration in problem-solving strategies. This article reports on a case study that focused on using the EDP to develop instructional materials by means of making a technological artifact to teach mathematical variables and functions at the secondary level. The five iterative stages of the EDP (design, make, test, infer, and iterate) were integrated into the development of the course materials. Data was collected from different sources: pre- and post-questionnaires, as well as a working document dealing with pupils' understanding based on designing, making, testing, and simulating. Twenty-four grade seven (13 years old) students in Northern Quebec participated in the study. The findings of this study indicate that STEM activities have a positive impact not only on students' engagement in classroom activities but also on learning new mathematical concepts. Furthermore, STEM-focused activities have a significant effect on problem-solving skills development in an interdisciplinary approach. Based on the study's results, we can conclude, inter alia, that teachers should integrate STEM activities into their teaching practices to increase learning outcomes and attach more importance to STEM-focused activities to develop students' reflective thinking and hands-on skills.Keywords: engineering design process, motivation, stem, integration, variables, functions
Procedia PDF Downloads 895636 Summer STEM Institute in Environmental Science and Data Sciencefor Middle and High School Students at Pace University
Authors: Lauren B. Birney
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Summer STEM Institute for Middle and High School Students at Pace University The STEM Collaboratory NYC® Summer Fellows Institute takes place on Pace University’s New York City campus during July and provides the following key features for all participants: (i) individual meetings with Pace faculty to discuss and refine future educational goals; (ii) mentorship, guidance, and new friendships with program leaders; and (iii) guest lectures from professionals in STEM disciplines and businesses. The Summer STEM Institute allows middle school and high school students to work in teams to conceptualize, develop, and build native mobile applications that teach and reinforce skills in the sciences and mathematics. These workshops enhance students’STEM problem solving techniques and teach advanced methods of computer science and engineering. Topics include: big data and analytics at the Big Data lab at Seidenberg, Data Science focused on social and environmental advancement and betterment; Natural Disasters and their Societal Influences; Algal Blooms and Environmental Impacts; Green CitiesNYC; STEM jobs and growth opportunities for the future; renew able energy and sustainable infrastructure; and climate and the economy. In order to better align the existing Summer STEM, Institute with the CCERS model and expand the overall network, Pace is actively recruiting new content area specialists from STEM industries and private sector enterprises to participate in an enhanced summer institute in order to1) nurture student progress and connect summer learning to school year curriculum, 2) increase peer-to-peer collaboration amongst STEM professionals and private sector technologists, and 3) develop long term funding and sponsorship opportunities for corporate sector partners to support CCERS schools and programs directly.Keywords: environmental restoration science, citizen science, data science, STEM
Procedia PDF Downloads 865635 Biocompatible Chitosan Nanoparticles as an Efficient Delivery Vehicle for Mycobacterium Tuberculosis Lipids to Induce Potent Cytokines and Antibody Response through Activation of γδ T-Cells in Mice
Authors: Ishani Das, Avinash Padhi, Sitabja Mukherjee, Santosh Kar, Avinash Sonawane
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Activation of cell mediated and humoral immune responses to Mycobacterium tuberculosis (Mtb) are critical for protection. Herein, we show that mice immunized with Mtb lipid bound chitosan nanoparticles(NPs) induce secretion of prominent Th1 and Th2 cytokines in lymph node and spleen cells, and also induced significantly higher levels of IgG, IgG1, IgG2 and IgM in comparison to control mice measured by ELISA. Furthermore, significantly enhanced γδ-T cell activation was observed in lymph node cells isolated from mice immunized with Mtb lipid coated chitosan-NPs as compared to mice immunized with chitosan-NPs alone or Mtb lipid liposomes through flow cytometric analysis. Also, it was observed that in comparison to CD8+ cells, significantly higher CD4+ cells were present in both the lymph node and spleen cells isolated from mice immunized with Mtb lipid coated chitosan NP. In conclusion, this study represents a promising new strategy for efficient delivery of Mtb lipids using chitosan NPs to trigger enhanced cell mediated and antibody response against Mtb lipids.Keywords: antibody response, chitosan nanoparticles, cytokines, mycobacterium tuberculosis lipids
Procedia PDF Downloads 2805634 Studying the Anti-Cancer Effects of Thymoquinone on Tumor Cells Through Natural Killer Cells Activity
Authors: Nouf A. Aldarmahi, Nesrin I. Tarbiah, Nuha A. Alkhattabi, Huda F. Alshaibi
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Nigella sativa which is known as dark cumin is a well-known example for a widely applicable herbal medicine. Nigella sativa can be effective in a variety of diseases such as hypertension, diabetes, bronchitis, gastrointestinal upset, and cancer. The anticancer effect of Nigella sativa appeared to be mediated by immune-modulatory effect through stimulating human natural killer (NK) cells. This is a type of lymphocytes which is part of the innate immunity, also known as the first line of defense in the body against pathogens. This study investigated the effect of thymoquinone as a major component of Nigella sativa on the molecular cytotoxic pathway of NK cell and the role of thymoquinone therapeutic effect on NK cells. NK cells were cultured with breast tumor cells in different ways and cultured media was collected and the concentration of perforin, granzyme B and interferon-α were measured by ELISA. The cytotoxic effect of NK cells on breast tumor cells was enhanced in the presence of thymoquinone, with increased activity of perforin in NK cells. This improved anticancer effect of thymoquinone on breast cancer cells.Keywords: breast cancer, cancer cells, natural killer cells, thymoquinone
Procedia PDF Downloads 2445633 Determination of Agricultural Characteristics of Smooth Bromegrass (Bromus inermis Leyss) Lines under Konya Regional Conditions
Authors: Abdullah Özköse, Ahmet Tamkoç
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The present study was conducted to determine the yield and yield components of smooth bromegrass lines under the environmental conditions of the Konya region during the growing seasons between 2011 and 2013. The experiment was performed in the randomized complete block design (RCBD) with four replications. It was found that the selected lines had a statistically significant effect on all the investigated traits, except for the main stem length and the number of nodes in the main stem. According to the two-year average calculated for various parameters checked in the smooth bromegrass lines, the main stem length ranged from 71.6 cm to 79.1 cm, the main stem diameter from 2.12 mm from 2.70 mm, the number of nodes in the main stem from 3.2 to 3.7, the internode length from 11.6 cm to 18.9 cm, flag leaf length from 9.7 cm to 12.7 cm, flag leaf width from 3.58 cm to 6.04 mm, herbage yield from 221.3 kg da–1 to 354.7 kg da–1 and hay yield from 100.4 kg da–1 to 190.1 kg da–1. The study concluded that the smooth bromegrass lines differ in terms of yield and yield components. Therefore, it is very crucial to select suitable varieties of smooth bromegrass to obtain optimum yield.Keywords: semiarid region, smooth bromegrass, yield, yield components
Procedia PDF Downloads 2765632 Using Lysosomal Immunogenic Cell Death to Target Breast Cancer via Xanthine Oxidase/Micro-Antibody Fusion Protein
Authors: Iulianna Taritsa, Kuldeep Neote, Eric Fossel
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Lysosome-induced immunogenic cell death (LIICD) is a powerful mechanism of targeting cancer cells that kills circulating malignant cells and primes the host’s immune cells against future remission. Current immunotherapies for cancer are limited in preventing recurrence – a gap that can be bridged by training the immune system to recognize cancer neoantigens. Lysosomal leakage can be induced therapeutically to traffic antigens from dying cells to dendritic cells, which can later present those tumorigenic antigens to T cells. Previous research has shown that oxidative agents administered in the tumor microenvironment can initiate LIICD. We generated a fusion protein between an oxidative agent known as xanthine oxidase (XO) and a mini-antibody specific for EGFR/HER2-sensitive breast tumor cells. The anti-EGFR single domain antibody fragment is uniquely sourced from llama, which is functional without the presence of a light chain. These llama micro-antibodies have been shown to be better able to penetrate tissues and have improved physicochemical stability as compared to traditional monoclonal antibodies. We demonstrate that the fusion protein created is stable and can induce early markers of immunogenic cell death in an in vitro human breast cancer cell line (SkBr3). Specifically, we measured overall cell death, as well as surface-expressed calreticulin, extracellular ATP release, and HMGB1 production. These markers are consensus indicators of ICD. Flow cytometry, luminescence assays, and ELISA were used respectively to quantify biomarker levels between treated versus untreated cells. We also included a positive control group of SkBr3 cells dosed with doxorubicin (a known inducer of LIICD) and a negative control dosed with cisplatin (a known inducer of cell death, but not of the immunogenic variety). We looked at each marker at various time points after cancer cells were treated with the XO/antibody fusion protein, doxorubicin, and cisplatin. Upregulated biomarkers after treatment with the fusion protein indicate an immunogenic response. We thus show the potential for this fusion protein to induce an anticancer effect paired with an adaptive immune response against EGFR/HER2+ cells. Our research in human cell lines here provides evidence for the success of the same therapeutic method for patients and serves as the gateway to developing a new treatment approach against breast cancer.Keywords: apoptosis, breast cancer, immunogenic cell death, lysosome
Procedia PDF Downloads 1995631 Silica Nanoparticles Induced Oxidative Stress and Inflammation in MRC-5 Human Lung Fibroblasts
Authors: Anca Dinischiotu, Sorina Nicoleta Voicu
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Silica nanoparticles (SiO2-NPs) are widely used in consumer products such as paints, plastics, insulation materials, tires, concrete production, as well as in gene delivery systems and imaging procedures. Environmental human exposure to them occurs during utilization of these products, in a time-dependent manner, the uptake being by topic and inhalation route especially. SiO2-NPs enter cells and induce membrane damage, oxidative stress and inflammatory reactions in a concentration-dependent manner. In this study, MRC-5 cells (human fetal lung fibroblasts) were exposed to amorphous SiO2-NPs at a dose of 62.5 μg/ml for 24, 48 and 72 hours. The size distribution of NPs was a lognormal function, in the range 3-14 nm. A time-dependent decrease of total reduced glutathione concentration by 36%, 50%, and 78% and an increase of NO level by 62%, 32%, respectively 24% compared to control were noticed. An up-regulation of NF-kB expression by 20%, 50% respectively 10% and of Nrf-2 by 139%, 58%, and 16% compared to control after 24, 48 and 72 hours was noticed also. The expression of IL-1β, IL-6, IL-8, and COX-2 was up-regulated in a time-dependent manner. Also, the expression of MMP-2 and MMP-9 were down-regulated after 48 and 72 hours, whereas their activities raised in a time-dependent manner. Exposure of cells to NPs up-regulated the expression of inducible NO synthase, as previously was shown, and probably this is the reason for the increased level of NO, that can react with the thiol groups of reduced glutathione molecules, diminishing its concentration Nrf2 is a transcription factor translocated in nucleus, under oxidative stress, where downstream gene expression activates in order to modulate the adaptive intracellular response against oxidative stress. The cross-talk between Nrf2 and NF-kB activities regulates the inflammatory processes. The activation of NF-kB could activate up-regulation of IL-1β, IL-6, and IL-8. The increase of COX-2 expression could be correlated with IL-1β one. Also, probably in response to the pro-inflammatory cytokines, MMP-2 and MMP-9 were induced and activated. In conclusion, the exposure of MRC-5 cells to SiO2-NPs generated inflammation in a time-dependent manner.Keywords: inflammation, MRC-5 cells, oxidative stress, silica nanoparticles
Procedia PDF Downloads 1475630 Hydroxyapatite Based Porous Scaffold for Tooth Tissue Engineering
Authors: Pakize Neslihan Taslı, Alev Cumbul, Gul Merve Yalcın, Fikrettin Sahin
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A key experimental trial in the regeneration of large oral and craniofacial defects is the neogenesis of osseous and ligamentous interfacial structures. Currently, oral regenerative medicine strategies are unpredictable for repair of tooth supporting tissues destroyed as a consequence of trauma, chronic infection or surgical resection. A different approach combining the gel-casting method with Hydroxy Apatite HA-based scaffold and different cell lineages as a hybrid system leads to successively mimic the early stage of tooth development, in vitro. HA is widely accepted as a bioactive material for guided bone and tooth regeneration. In this study, it was reported that, HA porous scaffold preparation, characterization and evaluation of structural and chemical properties. HA is the main factor that exists in tooth and it is in harmony with structural, biological, and mechanical characteristics. Here, this study shows mimicking immature tooth at the late bell stage design and construction of HA scaffolds for cell transplantation of human Adipose Stem Cells (hASCs), human Bone Marrow Stem Cells (hBMSCs) and Gingival Epitelial cells for the formation of human tooth dentin-pulp-enamel complexes in vitro. Scaffold characterization was demonstrated by SEM, FTIR and pore size and density measurements. The biological contraction of dental tissues against each other was demonstrated by mRNA gene expressions, histopatologic observations and protein release profile by ELISA tecnique. The tooth shaped constructs with a pore size ranging from 150 to 300 µm arranged by gathering right amounts of materials provide interconnected macro-porous structure. The newly formed tissue like structures that grow and integrate within the HA designed constructs forming tooth cementum like tissue, pulp and bone structures. These findings are important as they emphasize the potential biological effect of the hybrid scaffold system. In conclusion, this in vitro study clearly demonstrates that designed 3D scaffolds shaped as a immature tooth at the late bell stage were essential to form enamel-dentin-pulp interfaces with an appropriate cell and biodegradable material combination. The biomimetic architecture achieved here is providing a promising platform for dental tissue engineering.Keywords: tooth regeneration, tissue engineering, adipose stem cells, hydroxyapatite tooth engineering, porous scaffold
Procedia PDF Downloads 2345629 Prospects of Regenerative Medicine with Human Allogeneic Adipose Tissue-Derived Mesenchymal Stem Cell Sheets: Achievements and Future Outlook in Clinical Trials for Myopic Chorioretinal Atrophy
Authors: Norimichi Nagano, Yoshio Hirano, Tsutomu Yasukawa
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Mesenchymal stem cells are thought to confer neuroprotection, facilitate tissue regeneration and exert their effects on retinal degenerative diseases, however, adverse events such as proliferative vitreoretinopathy and preretinal membrane disease associated with cell suspension transplantation have also been reported. We have recently developed human (allogeneic) adipose tissue-derived mesenchymal stem cell (adMSC) sheets through our proprietary sheet transformation technique, which could potentially mitigate these adverse events. To clarify the properties of our adMSC sheets named PAL-222, we performed in vitro studies such as viability testing, cytokine secretions by ELISA, immunohistochemical study, and migration assay. The viability of the cells exceeded 70%. Vascular Endothelial Growth Factor (VEGF) and Pigment Epithelium-Derived Factor (PEDF), which are quite important cytokines for the retinal area, were observed. PAL-222 expressed type I collagen, a strength marker, type IV collagen, a marker of the basement membrane, and elastin, an elasticity marker. Finally, the migration assay was performed and showed negative, which means that PAL-222 is stably kept in the topical area and does not come to pieces. Next, to evaluate the efficacy in vivo, we transplanted PAL-222 into the subretinal space of the eye of Royal College of Surgeons rats with congenital retinal degeneration and assessed it for three weeks after transplantation. We confirmed that PAL-222 suppressed the decrease in the thickness of the outer nuclear layer, which means that the photoreceptor protective effect treated with PAL-222 was significantly higher than that in the sham group. (p < 0.01). This finding demonstrates that PAL-222 showed their retinoprotective effect in a model of congenital retinal degeneration. As the study suggested the efficacy of PAL-222 in both in vitro and in vivo studies, we are presently engaged in clinical trials of PAL-222 for myopic chorioretinal atrophy, which is one of the retinal degenerative diseases, for the purpose of regenerative medicine.Keywords: cell sheet, clinical trial, mesenchymal stem cell, myopic chorioretinal atrophy
Procedia PDF Downloads 945628 Breast Cancer Cellular Immunotherapies
Authors: Zahra Shokrolahi, Mohammad Reza Atashzar
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The goals of treating patients with breast cancer are to cure the disease, prolong survival, and improve quality of life. Immune cells in the tumor microenvironment have an important role in regulating tumor progression. The term of cellular immunotherapy refers to the administration of living cells to a patient; this type of immunotherapy can be active, such as a dendritic cell (DC) vaccine, in that the cells can stimulate an anti-tumour response in the patient, or the therapy can be passive, whereby the cells have intrinsic anti-tumour activity; this is known as adoptive cell transfer (ACT) and includes the use of autologous or allogeneic lymphocytes that may, or may not, be modified. The most important breast cancer cellular immunotherapies involving the use of T cells and natural killer (NK) cells in adoptive cell transfer, as well as dendritic cells vaccines. T cell-based therapies including tumour-infiltrating lymphocytes (TILs), engineered TCR-T cells, chimeric antigen receptor (CAR T cell), Gamma-delta (γδ) T cells, natural killer T (NKT) cells. NK cell-based therapies including lymphokine-activated killers (LAK), cytokine-induced killer (CIK) cells, CAR-NK cells. Adoptive cell therapy has some advantages and disadvantages some. TILs cell strictly directed against tumor-specific antigens but are inactive against tumor changes due to immunoediting. CIK cell have MHC-independent cytotoxic effect and also need concurrent high dose IL-2 administration. CAR T cell are MHC-independent; overcome tumor MHC molecule downregulation; potent in recognizing any cell surface antigen (protein, carbohydrate or glycolipid); applicable to a broad range of patients and T cell populations; production of large numbers of tumor-specific cells in a moderately short period of time. Meanwhile CAR T cells capable of targeting only cell surface antigens; lethal toxicity due to cytokine storm reported. Here we present the most popular cancer cellular immunotherapy approaches and discuss their clinical relevance referring to data acquired from clinical trials .To date, clinical experience and efficacy suggest that combining more than one immunotherapy interventions, in conjunction with other treatment options like chemotherapy, radiotherapy and targeted or epigenetic therapy, should guide the way to cancer cure.Keywords: breast cancer , cell therapy , CAR T cell , CIK cells
Procedia PDF Downloads 1315627 Conserved Stem-Loop Structure at the End of Short Interspersed Nuclear Elements (SINE) and Long Interspersed Nuclear Elements (LINE) Pairs of Different Species
Authors: Daria Grechishnikova, Maria Poptsova
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Transposable elements play an important role in the evolution of various species from bacteria to human. Long Interspersed Nuclear Elements (LINEs) and Short Interspersed Nuclear Elements (SINEs) are two major classes of retrotransposons that occupy a considerable part of any genome and their copy numbers can range form several hundreds to a million. Both LINEs and SINEs multiply through a copy-and-paste mechanism. LINEs encode proteins, which make them capable of self-propagation while SINEs are parasitic and require the machinery of LINEs to multiply. The mechanisms how LINE and SINE RNA is recognized by the LINE-encoded reverse transcriptase (RT) remain unclear. For some SINE-LINE pairs, it was shown that they share a common 3’-end with a stem-loop structure. Majority of the SINE-LINE pairs do not have a common 3’-end. Recently we have shown that in the human genome Alu-L1 pairs have structurally similar stem-loop structure at the 3’-end. Here we extended our analysis to a wide range of species and analyzed LINEs from 161 different species from Repbase and 217 SINE sequences from SINEBase. It appeared that all of the analyzed sequences contained stem-loop structures at the 3’-end. Here we conclude that it is very likely that a common evolutionary mechanism of transposon RNA recognition requires the presence of stem-loop structures at their 3’-end.Keywords: LINE, SINE, mechanisms of retrotransposition, retrotransposons, stem-loop, stem-loop structures, transposons
Procedia PDF Downloads 3535626 Single-Molecule Optical Study of Cholesterol-Mediated Dimerization Process of EGFRs in Different Cell Lines
Authors: Chien Y. Lin, Jung Y. Huang, Leu-Wei Lo
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A growing body of data reveals that the membrane cholesterol molecules can alter the signaling pathways of living cells. However, the understanding about how membrane cholesterol modulates receptor proteins is still lacking. Single-molecule tracking can effectively probe into the microscopic environments and thermal fluctuations of receptor proteins in a living cell. In this study we applies single-molecule optical tracking on ligand-induced dimerization process of EGFRs in the plasma membranes of two cancer cell lines (HeLa and A431) and one normal endothelial cell line (MCF12A). We tracked individual EGFR and dual receptors, diffusing in a correlated manner in the plasma membranes of live cells. We developed an energetic model by integrating the generalized Langevin equation with the Cahn-Hilliard equation to help extracting important information from single-molecule trajectories. From the study, we discovered that ligand-bound EGFRs move from non-raft areas into lipid raft domains. This ligand-induced motion is a common behavior in both cancer and normal cells. By manipulating the total amount of membrane cholesterol with methyl-β-cyclodextrin and the local concentration of membrane cholesterol with nystatin, we further found that the amount of cholesterol can affect the stability of EGFR dimers. The EGFR dimers in the plasma membrane of normal cells are more sensitive to the local concentration changes of cholesterol than EGFR dimers in the cancer cells. Our method successfully captures dynamic interactions of receptors at the single-molecule level and provides insight into the functional architecture of both the diffusing EGFR molecules and their local cellular environment.Keywords: membrane proteins, single-molecule tracking, Cahn-Hilliard equation, EGFR dimers
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