Search results for: adipose mesenchymal stem cells
3261 Brewing in a Domestic Refrigerator Using Freeze-Dried Raw Materials
Authors: Angelika-Ioanna Gialleli, Gousi Mantha, Maria Kanellaki, Bekatorou Argyro, Athanasios Koutinas
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In this study, a new brewing technology with dry raw materials is proposed with potential application in home brewing. Bio catalysts were prepared by immobilization of the psychrotolerant yeast strain Saccharomyces cerevisiae AXAZ-1 on tubular cellulose. Both the word and the biocatalysts were freeze-dried without any cryoprotectants and used for low temperature brewing. The combination of immobilization and freeze-drying techniques was applied successfully, giving a potential for supplying breweries with preserved and ready-to-use immobilized cells. The effect of wort sugar concentration (7°, 8.5°, 10°Be), temperature (2, 5, 7° C) and carrier concentration (5, 10, 20 g/L) on fermentation kinetics and final product quality (volatiles, colour, polyphenols, bitterness) was assessed. The same procedure was repeated with free cells for comparison of the results. The results for immobilized cells were better compared to free cells regarding fermentation kinetics and organoleptic characteristics.Keywords: brewing, tubular cellulose, low temperature, biocatalyst
Procedia PDF Downloads 3233260 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 2803259 Students’ Perception of Effort and Emotional Costs in Chemistry Courses
Authors: Guizella Rocabado, Cassidy Wilkes
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It is well known that chemistry is one of the most feared courses in college. Although many students enjoy learning about science, most of them perceive that chemistry is “too difficult”. These perceptions of chemistry result in many students not considering Science, Technology, Engineering, and Mathematics (STEM) majors because they require chemistry courses. Ultimately, these perceptions are also thought to be related to high attrition rates of students who begin STEM majors but do not persist. Students perceived costs of a chemistry class can be many, such as task effort, loss of valued alternatives, emotional, and others. These costs might be overcome by students’ interests and goals, yet the level of perceived costs might have a lasting impact on the students’ overall perception of chemistry and their desire to pursue chemistry and other STEM careers in the future. In this mixed methods study, we investigated task effort and emotional cost, as well as a mastery or performance goal orientation, and the impact these constructs may have on achievement in general chemistry classrooms. Utilizing cluster analysis as well as student interviews, we investigated students’ profiles of perceived cost and goal orientation as it relates to their final grades. Our results show that students who are well prepared for general chemistry, such as those who have taken chemistry in high school, display less negative perceived costs and thus believe they can master the material more fully. Other interesting results have also emerged from this research, which has the potential to have an impact on future instruction of these courses.Keywords: chemistry education, motivation, affect, perceived costs, goal orientations
Procedia PDF Downloads 903258 Experimental Research of Canine Mandibular Defect Construction with the Controlled Meshy Titanium Alloy Scaffold Fabricated by Electron Beam Melting Combined with BMSCs-Encapsulating Chitosan Hydrogel
Authors: Wang Hong, Liu Chang Kui, Zhao Bing Jing, Hu Min
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Objection We observed the repairment effection of canine mandibular defect with meshy Ti6Al4V scaffold fabricated by electron beam melting (EBM) combined with bone marrow mesenchymal stem cells (BMMSCs) encapsulated in chitosan hydrogel. Method Meshy titanium scaffolds were prepared by EBM of commercial Ti6Al4V power. The length of scaffolds was 24 mm, the width was 5 mm and height was 8mm. The pore size and porosity were evaluated by scanning electron microscopy (SEM). Chitosan /Bio-Oss hydrogel was prepared by chitosan, β- sodium glycerophosphate and Bio-Oss power. BMMSCs were harvested from canine iliac crests. BMMSCs were seeded in titanium scaffolds and encapsulated in Chitosan /Bio-Oss hydrogel. The validity of BMMSCs was evaluated by cell count kit-8 (CCK-8). The osteogenic differentiation ability was evaluated by alkaline phosphatase (ALP) activity and gene expression of OC, OPN and CoⅠ. Combination were performed by injecting BMMSCs/ Chitosan /Bio-Oss hydrogel into the meshy Ti6Al4V scaffolds and solidified. 24 mm long box-shaped bone defects were made at the mid-portion of mandible of adult beagles. The defects were randomly filled with BMMSCs/ Chitosan/Bio-Oss + titanium, Chitosan /Bio-Oss+titanium, titanium alone. Autogenous iliac crests graft as control group in 3 beagles. Radionuclide bone imaging was used to monitor the new bone tissue at 2, 4, 8 and 12 weeks after surgery. CT examination was made on the surgery day and 4 weeks, 12 weeks and 24 weeks after surgery. The animals were sacrificed in 4, 12 and 24 weeks after surgery. The bone formation were evaluated by histology and micro-CT. Results: The pores of the scaffolds was interconnected, the pore size was about 1 mm, the average porosity was about 76%. The pore size of the hydrogel was 50-200μm and the average porosity was approximately 90%. The hydrogel were solidified under the condition of 37℃in 10 minutes. The validity and the osteogenic differentiation ability of BMSCs were not affected by titanium scaffolds and hydrogel. Radionuclide bone imaging shown an increasing tendency of the revascularization and bone regeneration was observed in all the groups at 2, 4, 8 weeks after operation, and there were no changes at 12weeks.The tendency was more obvious in the BMMSCs/ Chitosan/Bio-Oss +titanium group and autogenous group. CT, Micro-CT and histology shown that new bone formed increasingly with the time extend. There were more new bone regenerated in BMMSCs/ Chitosan /Bio-Oss + titanium group and autogenous group than the other two groups. At 24 weeks, the autogenous group was achieved bone union. The BMSCs/ Chitosan /Bio-Oss group was seen extensive new bone formed around the scaffolds and more new bone inside of the central pores of scaffolds than Chitosan /Bio-Oss + titanium group and titanium group. The difference was significantly. Conclusion: The titanium scaffolds fabricated by EBM had controlled porous structure, good bone conduction and biocompatibility. Chitosan /Bio-Oss hydrogel had injectable plasticity, thermosensitive property and good biocompatibility. The meshy Ti6Al4V scaffold produced by EBM combined BMSCs encapsulated in chitosan hydrogel had good capacity on mandibular bone defect repair.Keywords: mandibular reconstruction, tissue engineering, electron beam melting, titanium alloy
Procedia PDF Downloads 4453257 A Green Analytical Curriculum for Renewable STEM Education
Authors: Mian Jiang, Zhenyi Wu
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We have incorporated green components into existing analytical chemistry curriculum with the aims to present a more environment benign approach in both teaching laboratory and undergraduate research. These include the use of cheap, sustainable, and market-available material; minimized waste disposal, replacement of non-aqueous media; and scale-down in sample/reagent consumption. Model incorporations have covered topics in quantitative chemistry as well as instrumental analysis, lower division as well as upper level, and research in traditional titration, spectroscopy, electrochemical analysis, and chromatography. The green embedding has made chemistry more daily life relevance, and application focus. Our approach has the potential to expand into all STEM fields to make renewable, high-impact education experience for undergraduate students.Keywords: green analytical chemistry, pencil lead, mercury, renewable
Procedia PDF Downloads 3393256 Neural Nets Based Approach for 2-Cells Power Converter Control
Authors: Kamel Laidi, Khelifa Benmansour, Ouahid Bouchhida
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Neural networks-based approach for 2-cells serial converter has been developed and implemented. The approach is based on a behavioural description of the different operating modes of the converter. Each operating mode represents a well-defined configuration, and for which is matched an operating zone satisfying given invariance conditions, depending on the capacitors' voltages and the load current of the converter. For each mode, a control vector whose components are the control signals to be applied to the converter switches has been associated. Therefore, the problem is reduced to a classification task of the different operating modes of the converter. The artificial neural nets-based approach, which constitutes a powerful tool for this kind of task, has been adopted and implemented. The application to a 2-cells chopper has allowed ensuring efficient and robust control of the load current and a high capacitors voltages balancing.Keywords: neural nets, control, multicellular converters, 2-cells chopper
Procedia PDF Downloads 8343255 Control of IL-23 Release in Dendritic Cells Protects Mice from Imiquimod-Induced Psoriasis
Authors: Xingxin Wu, Fenli Shao, Tao Tan, Yang Tan, Yang Sun, Qiang Xu
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Psoriasis is a chronic inflammatory skin disease that affects about 2% of the world's population. IL-23 signaling plays a key role in the pathogenesis of psoriasis. Control of IL-23 release by small molecule compounds during developing psoriasis has not been well established. Here, we show that compound 1, a small molecule nature product, protected mice from imiquimod-induced psoriasis with improved skin lesions, reduced skin thickness, and reduced IL-23 mRNA expression in the skin tissue. FACS results showed compound 1 reduced the number of dendritic cells in the skin. Interestingly, compound 1 was not able to ameliorate IL-23-induced psoriasis-like skin inflammation in mice. Further, compound 1 inhibited MyD88-dependent IL-23 mRNA expression induced by LPS, CpG and imiquimod in BMDC cells, but not MyD88-independent CD80 and CD86 expression induced by LPS. The methods included real-time PCR, western blot, H & E staining, FACS and ELISA et al. In conclusion, compound 1 regulates MyD88-dependent signaling to control IL-23 release in dendritic cells, which improves imiquimod-induced psoriasis.Keywords: dendritic cells, IL-23, toll-like receptor signaling, psoriasis
Procedia PDF Downloads 6453254 Non Interferometric Quantitative Phase Imaging of Yeast Cells
Authors: P. Praveen Kumar, P. Vimal Prabhu, Renu John
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In biology most microscopy specimens, in particular living cells are transparent. In cell imaging, it is hard to create an image of a cell which is transparent with a very small refractive index change with respect to the surrounding media. Various techniques like addition of staining and contrast agents, markers have been applied in the past for creating contrast. Many of the staining agents or markers are not applicable to live cell imaging as they are toxic. In this paper, we report theoretical and experimental results from quantitative phase imaging of yeast cells with a commercial bright field microscope. We reconstruct the phase of cells non-interferometrically based on the transport of intensity equations (TIE). This technique estimates the axial derivative from positive through-focus intensity measurements. This technique allows phase imaging using a regular microscope with white light illumination. We demonstrate nano-metric depth sensitivity in imaging live yeast cells using this technique. Experimental results will be shown in the paper demonstrating the capability of the technique in 3-D volume estimation of living cells. This real-time imaging technique would be highly promising in real-time digital pathology applications, screening of pathogens and staging of diseases like malaria as it does not need any pre-processing of samples.Keywords: axial derivative, non-interferometric imaging, quantitative phase imaging, transport of intensity equation
Procedia PDF Downloads 3843253 A Comparative Study on Biochar from Slow Pyrolysis of Corn Cob and Cassava Wastes
Authors: Adilah Shariff, Nurhidayah Mohamed Noor, Alexander Lau, Muhammad Azwan Mohd Ali
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Biomass such as corn and cassava wastes if left to decay will release significant quantities of greenhouse gases (GHG) including carbon dioxide and methane. The biomass wastes can be converted into biochar via thermochemical process such as slow pyrolysis. This approach can reduce the biomass wastes as well as preserve its carbon content. Biochar has the potential to be used as a carbon sequester and soil amendment. The aim of this study is to investigate the characteristics of the corn cob, cassava stem, and cassava rhizome in order to identify their potential as pyrolysis feedstocks for biochar production. This was achieved by using the proximate and elemental analyses as well as calorific value and lignocellulosic determination. The second objective is to investigate the effect of pyrolysis temperature on the biochar produced. A fixed bed slow pyrolysis reactor was used to pyrolyze the corn cob, cassava stem, and cassava rhizome. The pyrolysis temperatures were varied between 400 °C and 600 °C, while the heating rate and the holding time were fixed at 5 °C/min and 1 hour, respectively. Corn cob, cassava stem, and cassava rhizome were found to be suitable feedstocks for pyrolysis process because they contained a high percentage of volatile matter more than 80 mf wt.%. All the three feedstocks contained low nitrogen and sulphur content less than 1 mf wt.%. Therefore, during the pyrolysis process, the feedstocks give off very low rate of GHG such as nitrogen oxides and sulphur oxides. Independent of the types of biomass, the percentage of biochar yield is inversely proportional to the pyrolysis temperature. The highest biochar yield for each studied temperature is from slow pyrolysis of cassava rhizome as the feedstock contained the highest percentage of ash compared to the other two feedstocks. The percentage of fixed carbon in all the biochars increased as the pyrolysis temperature increased. The increment of pyrolysis temperature from 400 °C to 600 °C increased the fixed carbon of corn cob biochar, cassava stem biochar and cassava rhizome biochar by 26.35%, 10.98%, and 6.20% respectively. Irrespective of the pyrolysis temperature, all the biochars produced were found to contain more than 60 mf wt.% fixed carbon content, much higher than its feedstocks.Keywords: biochar, biomass, cassava wastes, corn cob, pyrolysis
Procedia PDF Downloads 2993252 Cervical Cell Classification Using Random Forests
Authors: Dalwinder Singh, Amandeep Verma, Manpreet Kaur, Birmohan Singh
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The detection of pre-cancerous changes using a Pap smear test of cervical cell is the important step for the early diagnosis of cervical cancer. The Pap smear test consists of a sample of human cells taken from the cervix which are analysed to detect cancerous and pre-cancerous stage of the given subject. The manual analysis of these cells is labor intensive and time consuming process which relies on expert cytotechnologist. In this paper, a computer assisted system for the automated analysis of the cervical cells has been proposed. We propose a morphology based approach to the nucleus detection and segmentation of the cytoplasmic region of the given single or multiple overlapped cell. Further, various texture and region based features are calculated from these cells to classify these into normal and abnormal cell. Experimental results on public available dataset show that our system has achieved satisfactory success rate.Keywords: cervical cancer, cervical tissue, mathematical morphology, texture features
Procedia PDF Downloads 5263251 Prolonged Synthesis of Chitin Polysaccharide from Chlorovirus System
Authors: Numfon Rakkhumkaew, Takeru Kawasaki, Makoto Fujie, Takashi Yamada
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Chlorella viruses or chloroviruses contain a gene that encodes a function for chitin synthesis, which is expressed early in viral infection to produce chitin polysaccharide, a polymer of β-1, 4-linked GlcNAc, on the outside of Chlorella cell wall. Interestingly, chlorovirus system is an eco-friendly system which converses CO2 and solar energy from the environment into useful materials. However, infected Chlorella cells are lysed at the final stage of viral infection, and this phenomenon is caused the breaking down of polysaccharide. To postpone the lysing period and prolong the synthesis of chitin polysaccharide on cells, the slow growing virus incorporated with aphidicolin treatment, an inhibitor of DNA synthesis, was investigated. In this study, a total of 25 virus isolates from water samples in Japan region were analyzed for CHS (the gene for CH synthase) gene by PCR (polymerase chain reaction). The accumulation and appearance of chitin polysaccharide on infected cells were detected by biotinylated chitin-binding proteins WGA (wheat germ agglutinin)-biotin for chitin in conjunction with avidin-Cy 2 or Cy 3 and investigated by fluorescence microscopy, observed as green or yellow fluorescence over the cell surface. Among all chlorovirus isolates, cells infected with CNF1 revealed the accumulation of chitin over the cell surface within 30 min p.i. and continued to accumulate on cells until 4 h p.i. before cell lyses which was 1.6 times longer accumulation period than cells infected with CVK2 (prototype virus). Furthermore, addition of aphidicolin could extend the chitin accumulation on cells infected with CNF1 until 8 h p.i. before cell lyses. Whereas, CVK2-infected cells treated with aphidicolin could prolong the chitin synthesis only for 6 h p.i. before cell lyses. Therefore, chitin synthesis by Chlorella-virus system could be prolonged by using slow-growing viral isolates and with aphidicolin.Keywords: chitin, chlorovirus, Chlorella virus, aphidicolin
Procedia PDF Downloads 2133250 Evaluation of the Synergistic Inhibition of Enterovirus 71 Infection by Interferon-α Coupled with Pleconaril in RD Cells
Authors: Wen-Yu Lin, Yi-Ching Chung, Tzyy-Rong Jinn
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It is well known that enterovirus 71 (EV71) causes recurring outbreaks of hand, foot and mouth disease (HFMD) and encephalitis leading to complications or death in young children. And, several HFMD of EV71 with high mortalities occurred in Asia countries, such as Malaysia (1997), Taiwan (1998) and China (2008). Thus, more effective antiviral drugs are needed to prevent or reduce EV71-related complications. As reported, interferon-α protects mice from lethal EV71 challenge by the modulation of innate immunity and then degrade enterovirus protease 3Cᵖʳᵒ. On the other side, pleconaril by targeting enterovirus VP1 protein and then block virus entry and attachment. Thus, the aim of this study was to evaluate the synergistic antiviral activity of interferon-α and pleconaril against enterovirus 71 infection. In a preliminary study showed that pleconaril at concentrations of 50, 100 and 300 µg/mL reduced EV71-induced CPE to 52.0 ± 2.5%, 40.2 ± 3.5% and 26.5 ± 1.5%, respectively, of that of the EV71-infected RD control cells (taken as 100%). Notably, 1000 IU/mL of interferon-α in combination with pleconaril at concentrations of 50, 100 and 300µg/mL suppressed EV71-induced CPE by 30.2 ± 3.8%, 16.5 ± 1.3% and 2.8 ± 2.0%, respectively, of that of the pleconaril alone treated with the infected RD cells. These results indicated that interferon-α 1000 IU/mL combination with pleconaril (50, 100 and 300µg/mL) inhibited EV71-induced CPE more effectively than treated with pleconaril alone in the infected RD cells.Keywords: enterovirus 71, interferon-α, pleconaril, RD cells
Procedia PDF Downloads 1403249 Protective Potential of Hyperhalophilic Diatoms Extract Against Lead Induced Oxidative Stress in Rats and Human HepG2 and HEK293 Cells Line
Authors: Wassim Guermazi, Saoussan Boukhris, Neila Annabi Trabelsi, Tarek Rebai, Alya Sellami-Kamoun, Habib Ayadi
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This work investigates the protective effects of the microalga Halamphora sp. extract (H. Ext) as a natural product on lead-intoxicated liver and kidney human cells in vitro and in vivo on rats wistar. HepG2 cells line derived from human hepatocellular carcinoma and HEK293 cells line derived from human embryonic kidney were used for the in vitro study. The analysis of the fatty acids methyl esters of the extract was performed by a GC/MS. Four groups of rats, each of which was composed of six animals, were used for the in vivo experiment. The pretreatment of HepG2 and HEK293 cells line with the extract (100 µg mL-1) significantly (p < 0.05) protected against cytotoxicity induced by lead exposure. In vivo, the biochemical parameters in serum, namely malondialdehyde level (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) activities, were measured in supernatants of organ homogenates. H. Ext was found to be rich in fatty acids, essentially palmitic and palmitoleic accounting respectively 29.46% and 42.07% of total fatty acids. Both in vitro and in vivo, the co-treatment with H. Ext allowed the protection of the liver and kidney cells structure, as well as the significant preservation of normal antioxidant and biochemical parameters in rats. Halamphora extract rich in fatty acids has been proven to be effective in protection against Pb-induced toxicity.Keywords: microalga extract, human cells line, fatty acid, lead exposure, oxidative stress, rats
Procedia PDF Downloads 883248 Analysis of Anti-Tuberculosis Immune Response Induced in Lungs by Intranasal Immunization with Mycobacterium indicus pranii
Authors: Ananya Gupta, Sangeeta Bhaskar
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Mycobacterium indicus pranii (MIP) is a saprophytic mycobacterium. It is a predecessor of M. avium complex (MAC). Whole genome analysis and growth kinetics studies have placed MIP in between pathogenic and non-pathogenic species. It shares significant antigenic repertoire with M. tuberculosis and have unique immunomodulatory properties. MIP provides better protection than BCG against pulmonary tuberculosis in animal models. Immunization with MIP by aerosol route provides significantly higher protection as compared to immunization by subcutaneous (s.c.) route. However, mechanism behind differential protection has not been studied. In this study, using mice model we have evaluated and compared the M.tb specific immune response in lung compartments (airway lumen / lung interstitium) as well as spleen following MIP immunization via nasal (i.n.) and s.c. route. MIP i.n. vaccination resulted in increased seeding of memory T cells (CD4+ and CD8+ T-cells) in the airway lumen. Frequency of CD4+ T cells expressing Th1 migratory marker (CXCR3) and activation marker (CD69) were also high in airway lumen of MIP i.n. group. Significantly high ex vivo secretion of cytokines- IFN-, IL-12, IL-17 and TNF- from cells of airway luminal spaces provides evidence of antigen-specific lung immune response, besides generating systemic immunity comparable to MIP s.c. group. Analysis of T cell response on per cell basis revealed that antigen specific T-cells of MIP i.n. group were functionally superior as higher percentage of these cells simultaneously secreted IFN-gamma, IL-2 and TNF-alpha cytokines as compared to MIP s.c. group. T-cells secreting more than one of the cytokines simultaneously are believed to have robust effector response and crucial for protection, compared with single cytokine secreting T-cells. Adoptive transfer of airway luminal T-cells from MIP i.n. group into trachea of naive B6 mice revealed that MIP induced CD8 T-cells play crucial role in providing long term protection. Thus the study demonstrates that MIP intranasal vaccination induces M.tb specific memory T-cells in the airway lumen that results in an early and robust recall response against M.tb infection.Keywords: airway lumen, Mycobacterium indicus pranii, Th1 migratory markers, vaccination
Procedia PDF Downloads 1873247 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 3743246 A New Alpha-Amylase Inhibitor Isolated from the Stem Bark of Anthocleista Djalonensis
Authors: Oseyemi O. Olubomehin, Edith O. Ajaiyeoba, Kio A. Abo, Eleonora D. Goosen
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Diabetes is a major degenerative disease of global concern and it is the third most lethal disease of mankind, accounting for about 3.2 million deaths annually. Lowering postprandial hyperglycemia by inhibition of carbohydrate hydrolyzing enzyme such as alpha-amylase is one of the therapeutic approaches to treat Type 2 Diabetes. Alpha-amylase inhibitors from plants have been found to be effective in managing postprandial hyperglycemia. In continuation of our anti-diabetic activities of this plant, bioassay-guided fractionation and isolation using 0.1-1.0 mg/mL furnished djalonenol, a monoterpene diol with a significant 53.7% α-amylase inhibition (p<0.001) from the stem bark which was comparable to acarbose which gave a 54.9% inhibition. Spectral characterization using Infra-red, Gas Chromatogrphy-Mass spectrometry, 1D and 2D NMR of the isolated compound was done to elucidate the structure of the compound.Keywords: alpha-amylase inhibitor, hyperglycemia, postprandial, diabetes
Procedia PDF Downloads 4583245 A Strategic Sustainability Analysis of Electric Vehicles in EU Today and Towards 2050
Authors: Sven Borén, Henrik Ny
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Ambitions within the EU for moving towards sustainable transport include major emission reductions for fossil fuel road vehicles, especially for buses, trucks, and cars. The electric driveline seems to be an attractive solution for such development. This study first applied the Framework for Strategic Sustainable Development to compare sustainability effects of today’s fossil fuel vehicles with electric vehicles that have batteries or hydrogen fuel cells. The study then addressed a scenario were electric vehicles might be in majority in Europe by 2050. The methodology called Strategic Lifecycle Assessment was first used, were each life cycle phase was assessed for violations against sustainability principles. This indicates where further analysis could be done in order to quantify the magnitude of each violation, and later to create alternative strategies and actions that lead towards sustainability. A Life Cycle Assessment of combustion engine cars, plug-in hybrid cars, battery electric cars and hydrogen fuel cell cars was then conducted to compare and quantify environmental impacts. The authors found major violations of sustainability principles like use of fossil fuels, which contribute to the increase of emission related impacts such as climate change, acidification, eutrophication, ozone depletion, and particulate matters. Other violations were found, such as use of scarce materials for batteries and fuel cells, and also for most life cycle phases for all vehicles when using fossil fuel vehicles for mining, production and transport. Still, the studied current battery and hydrogen fuel cell cars have less severe violations than fossil fuel cars. The life cycle assessment revealed that fossil fuel cars have overall considerably higher environmental impacts compared to electric cars as long as the latter are powered by renewable electricity. By 2050, there will likely be even more sustainable alternatives than the studied electric vehicles when the EU electricity mix mainly should stem from renewable sources, batteries should be recycled, fuel cells should be a mature technology for use in vehicles (containing no scarce materials), and electric drivelines should have replaced combustion engines in other sectors. An uncertainty for fuel cells in 2050 is whether the production of hydrogen will have had time to switch to renewable resources. If so, that would contribute even more to a sustainable development. Except for being adopted in the GreenCharge roadmap, the authors suggest that the results can contribute to planning in the upcoming decades for a sustainable increase of EVs in Europe, and potentially serve as an inspiration for other smaller or larger regions. Further studies could map the environmental effects in LCA further, and include other road vehicles to get a more precise perception of how much they could affect sustainable development.Keywords: strategic, electric vehicles, sustainability, LCA
Procedia PDF Downloads 3853244 Survey of the Effect of the Probiotic Bacterium Lactobacillus plantarum and Streptococcus mutans on Casp3, AKT/PTEN, and MAPK Signaling Pathways at Co-Culture with KB Oral Cancer Cell Line and HUVEC Cells
Authors: Negar Zaheddoust, Negin Zaheddoust, Abbas Asoudeh-Fard
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Probiotic bacteria have been employed as a novel and less side-effect strategy for anticancer therapy. Since the oral cavity is a host for probiotic and pathogen bacteria to colonize, more investigation is needed to evaluate the effectiveness of this novel adjunctive treatment for oral cancer. We considered Lactobacillus plantarum as a probiotic and Streptococcus mutans as a pathogen bacterium in our study. The aim of this study is to examine the effect of Lactobacillus plantarum and Streptococcus mutans on Casp3, AKT / PTEN, and MAPK signaling pathway, which is involved in apoptosis or survival of oral cancer KB cells. On the other hand, to study the effects of these bacteria on normal cells, we used HUVEC cells. The KB and HUVEC cell lines were co-cultured with Lactobacillus plantarum and Streptococcus mutans isolated from traditional Iranian dairy and dental plaque, respectively. The growth-inhibitory effects of these two bacteria on KB and HUVEC cells were determined by (3-(4, 5-dimethylthiazolyl-2)-2,5diphenyltetrazolium bromide) MTT assay. MTT results demonstrated that the proliferation of KB cells was affected in a time, dose, and strain-dependent manner. In the following, the examination of induced apoptosis or necrosis in co-cultured KB cells with the best IC50 concentration of the Lactobacillus plantarum and Streptococcus mutans will be analyzed by FACS flow cytometry, and the changes in gene expression of Casp3, AKT / PTEN, MAPK genes will be evaluated using real-time polymerase chain reaction.Keywords: cancer therapy, induced apoptosis, oral cancer, probiotics
Procedia PDF Downloads 2483243 Yield Loss in Maize Due to Stem Borers and Their Integrated Management
Authors: C. P. Mallapur, U. K. Hulihalli, D. N. Kambrekar
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Maize (Zea mays L.) an important cereal crop in the world has diversified uses including human consumption, animal feed, and industrial uses. A major constraint in low productivity of maize in India is undoubtedly insect pests particularly two species of stem borers, Chilo partellus (Swinhoe) and Sesamia inferens (Walker). The stem borers cause varying level of yield losses in different agro-climate regions (25.7 to 80.4%) resulting in a huge economic loss to the farmers. Although these pests are rather difficult to manage, efforts have been made to combat the menace by using effective insecticides. However, efforts have been made in the present study to integrate various possible approaches for sustainable management of these borers. Two field experiments were conducted separately during 2016-17 at Main Agricultural Research Station, University of Agricultural Sciences, Dharwad, Karnataka, India. In the first experiment, six treatments were randomized in RBD. The insect eggs at pinhead stage (@ 40 eggs/plant) were stapled to the under surface of leaves covering 15-20 % of plants in each plot after 15 days of sowing. The second experiment was planned with nine treatments replicated thrice. The border crop with NB -21 grass was planted all around the plots in the specific treatments while, cowpea intercrop (@6:1-row proportion) was sown along with the main crop and later, the insecticidal spray with chlorantraniliprole and nimbecidine was taken upon need basis in the specific treatments. The results indicated that the leaf injury and dead heart incidence were relatively more in the treatments T₂ and T₄ wherein, no insect control measures were made after the insect release (58.30 & 40.0 % leaf injury and 33.42 and 25.74% dead heart). On the contrary, these treatments recorded higher stem tunneling (32.4 and 24.8%) and resulted in lower grain yield (17.49 and 26.79 q/ha) compared to 29.04, 32.68, 40.93 and 46.38 q/ha recorded in T₁, T₃, T₅ and T₆ treatments, respectively. A maximum yield loss of 28.89 percent was noticed in T₂ followed by 19.59 percent in T₄ where no sprays were imposed. The data on integrated management trial revealed the lowest stem borer damage (19.28% leaf injury and 1.21% dead heart) in T₅ (seed treatment with thiamethoxam 70FS @ 8ml/kg seed + cow intercrop along with nimbecidine 0.03EC @ 5.0 ml/l and chlorantraniliprole 18.5SC spray @ 0.2 ml/l). The next best treatment was T₆ (ST+ NB-21 borer with nimbecidine and chlorantraniliprole spray) with 21.3 and 1.99 percent leaf injury and dead heart incidence, respectively. These treatments resulted in highest grain yield (77.71 and 75.53 q/ha in T₅ and T₆, respectively) compared to the standard check, T₁ (ST+ chlorantraniliprole spray) wherein, 27.63 percent leaf injury and 3.68 percent dead heart were noticed with 60.14 q/ha grain yield. The stem borers can cause yield loss up to 25-30 percent in maize which can be well tackled by seed treatment with thiamethoxam 70FS @ 8ml/kg seed and sowing the crop along with cowpea as intercrop (6:1 row proportion) or NB-21 grass as border crop followed by application of nimbecidine 0.03EC @ 5.0 ml/l and chlorantraniliprole 18.5SC @ 0.2 ml/l on need basis.Keywords: Maize stem borers, Chilo partellus, Sesamia inferens, crop loss, integrated management
Procedia PDF Downloads 1763242 Comparative Antibacterial Property of Matured Trunk and Stem Bark Extract of Tamarindus indica L., Preformulation, Development and Quality Control of Cream
Authors: A. M. T. Jacinto, M.O. Osi
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Tamarind has various medicinal properties among which is its antibacterial property. Its bark contains saponins, alkaloids, sesquiterpenes and tannins. It is rich in phlobapenes which is responsible for antibacterial property. The objective of the study was to determine which bark will produce the highest antibacterial property, develop it into a topical cream and evaluate its quality and characteristics. Powdered barks of Tamarind were extracted by soxhlet method using 70% acetone. Stem bark produced a higher yield than trunk bark (5.85 g vs. 4.73 g). It was found that the trunk bark was more sensitive than stem bark to microorganisms namely Staphylococcus aureus, Corynebacterium minutissimum, and Streptococcus spp. Sensitivity of trunk bark can be attributed to a more developed phytoconstituents. Dermal sensitization test on both sexes of rabbits using the following concentrations: 100%, 40% and 20% of extract showed that Tamarind has no irritating property and therefore safe for formulation into an antibacterial cream. Excipients used for formulation such as methyl paraben, propyl paraben, stearyl alcohol and white petrolatum were compatible with the Tamarind acetone extract through Differential Scanning Calorimetry except sodium lauryl sulfate that exhibited crystallization when subjected at 200˚C. The method of manufacture used in cream is fusion, therefore strict compliance of processing temperature should be observed to prevent polymorphism. Quality control tests of formulated cream based on USP 30 and Philippine Pharmacopeia were satisfactory.Keywords: antibacterial, differential scanning calorimetry, tannins, dermal sensitization
Procedia PDF Downloads 4863241 Ancelim: Health System Restoration Protocol for Cancer Patients
Authors: Mark Berry
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A number of studies have identified several factors involved in the malignant progression of cancer cells. The Primary modulator in driving inflammation to these transformed cells has been identified as the transcription factor known as nuclear factor-κB. This essential regulator of inflammation and the development of cancer, combined with a microenvironment of inflammation and signaling molecules, plays a major role in the malignant progression of cancer, and this progression is the result of the mutagenic predisposition of persistent substances that combat infection at tumor sites and other areas of chronic inflammation. Inflammation-induced tumors, and their inflammatory cells and regulators may be the primary source of metastasis of tumor cells through angiogenesis. Previous research on cytokines and chemokines, including their downstream targets, has been the focus of the cancer/inflammation connection. The identification of the biological mechanisms of other proteins vital to the inflammation cascade and their interactions are crucial to novel and effective therapeutic protocols for the treatment of inflammation-induced cancers. The Ancelim HSRP Protocol is just such a therapeutic intervention.Keywords: ancelim, cancer, inflammation, tumor
Procedia PDF Downloads 5453240 Physiological Normoxia and Cellular Adhesion of Diffuse Large B-Cell Lymphoma Primary Cells: Real-Time PCR and Immunohistochemistry Study
Authors: Kamila Duś-Szachniewicz, Kinga M. Walaszek, Paweł Skiba, Paweł Kołodziej, Piotr Ziółkowski
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Cell adhesion is of fundamental importance in the cell communication, signaling, and motility, and its dysfunction occurs prevalently during cancer progression. The knowledge of the molecular and cellular processes involved in abnormalities in cancer cells adhesion has greatly increased, and it has been focused mainly on cellular adhesion molecules (CAMs) and tumor microenvironment. Unfortunately, most of the data regarding CAMs expression relates to study on cells maintained in standard oxygen condition of 21%, while the emerging evidence suggests that culturing cells in ambient air is far from physiological. In fact, oxygen in human tissues ranges from 1 to 11%. The aim of this study was to compare the effects of physiological lymph node normoxia (5% O2), and hyperoxia (21% O2) on the expression of cellular adhesion molecules of primary diffuse large B-cell lymphoma cells (DLBCL) isolated from 10 lymphoma patients. Quantitative RT-PCR and immunohistochemistry were used to confirm the differential expression of several CAMs, including ICAM, CD83, CD81, CD44, depending on the level of oxygen. Our findings also suggest that DLBCL cells maintained at ambient O2 (21%) exhibit reduced growth rate and migration ability compared to the cells growing in normoxia conditions. Taking into account all the observations, we emphasize the need to identify the optimal human cell culture conditions mimicking the physiological aspects of tumor growth and differentiation.Keywords: adhesion molecules, diffuse large B-cell lymphoma, physiological normoxia, quantitative RT-PCR
Procedia PDF Downloads 2783239 Genetic Analysis of Rust Resistance Genes in Global Wheat
Authors: Aktar-Uz-Zaman, M. Tuhina-Khatun, Mohamed Hanafi Musa
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Three rust diseases: leaf (brown) rust caused by Puccinia triticina Eriks, stripe (yellow) rust caused by Puccinia striiformis West, and stem (black) rust caused by Puccinia graminis f. sp. tritici are economically important diseases of wheat in world wide. Yield loss due to leaf rust is 40% in susceptible cultivars. Yield losses caused by the stem rust pathogens in the mid of 20 century reached 20-30% in Eastern and Central Europe and the most virulent stem rust race Ug99 emerged first in Uganda and after that in Kenya, Ethiopia, Yemen, in the Middle East and South Asia. Yield losses were estimated up to 100%, whereas, up to 80% have been reported in Kenya during 1999. In case of stripe rust, severity level has been recorded 60% - 70% as compared to 100% severity of susceptible check in disease screening nurseries in Kenya. Improvement of resistant varieties or cultivars is the sustainable, economical and environmentally friendly approaches for increasing the global wheat production to suppress the rust diseases. More than 68 leaf rust, 49 stripe rust and 53 stem rust resistance genes have been identified in the global wheat cultivars or varieties using different molecular breeding approaches. Among these, Lr1, Lr9, Lr10, Lr19, Lr21, Lr24, Lr25, Lr28, Lr29, Lr34, Lr35, Lr37, Lr39, Lr47, Lr51, Lr3bg, Lr18, Lr40, Lr46, and Lr50 leaf rust resistance genes have been identified by using molecular, enzymatic and microsatellite markers from African, Asian, European cultivars of hexaploid wheat (Triticum aestivum), durum wheat and diploid wheat species. These genes are located on 20, of the 21 chromosomes of hexaploid wheat. Similarly, Sr1, Sr2, Sr24, and Sr3, Sr31 stem rust resistance genes have been recognized from wheat cultivars of Pakistan, India, Kenya, and Uganda etc. A race of P. striiformis (stripe rust) Yr9, Yr18, and Yr29 was first observed in East Africa, Italy, Pakistan and India wheat cultivars. These stripe rust resistance genes are located on chromosomes 1BL, 4BL, 6AL, 3BS and 6BL in bread wheat cultivars. All these identified resistant genes could be used for notable improvement of susceptible wheat cultivars in the future.Keywords: hexaploid wheat, resistance genes, rust disease, triticum aestivum
Procedia PDF Downloads 4813238 A Double-Blind, Randomized, Controlled Trial on N-Acetylcysteine for the Prevention of Acute Kidney Injury in Patients Undergoing Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Sara Ataei, Molouk Hadjibabaie, Amirhossein Moslehi, Maryam Taghizadeh-Ghehi, Asieh Ashouri, Elham Amini, Kheirollah Gholami, Alireza Hayatshahi, Mohammad Vaezi, Ardeshir Ghavamzadeh
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Acute kidney injury (AKI) is one of the complications of hematopoietic stem cell transplantation and is associated with increased mortality. N-acetylcysteine (NAC) is a thiol compound with antioxidant and vasodilatory properties that has been investigated for the prevention of AKI in several clinical settings. In the present study, we evaluated the effects of intravenous NAC on the prevention of AKI in allogeneic hematopoietic stem cell transplantation patients. A double-blind randomized placebo-controlled trial was conducted, and 80 patients were recruited to receive 100 mg/kg/day NAC or placebo as intermittent intravenous infusion from day -6 to day +15. AKI was determined on the basis of the Risk-Injury-Failure-Loss-Endstage renal disease and AKI Network criteria as the primary outcome. We assessed urine neutrophil gelatinase-associated lipocalin (uNGAL) on days -6, -3, +3, +9, and +15 as the secondary outcome. Moreover, transplant-related outcomes and NAC adverse reactions were evaluated during the study period. Statistical analysis was performed using appropriate parametric and non-parametric methods including Kaplan–Meier for AKI and generalized estimating equation for uNGAL. At the end of the trial, data from 72 patients were analyzed (NAC: 33 patients and placebo: 39 patients). Participants of each group were not different considering baseline characteristics. AKI was observed in 18% of NAC recipients and 15% of placebo group patients, and the occurrence pattern was not significantly different (p = 0.73). Moreover, no significant difference was observed between groups for uNGAL measures (p = 0.10). Transplant-related outcomes were similar for both groups, and all patients had successful engraftment. Three patients did not tolerate NAC because of abdominal pain, shortness of breath and rash with pruritus and were dropped from the intervention group before transplantation. However, the frequency of adverse reactions was not significantly different between groups. In conclusion, our findings could not show any clinical benefits from high-dose NAC particularly for AKI prevention in allogeneic hematopoietic stem cell transplantation patients.Keywords: acute kidney injury, N-acetylcysteine, hematopoietic stem cell transplantation, urine neutrophil gelatinase-associated lipocalin, randomized controlled trial
Procedia PDF Downloads 4333237 Microfluidic Based High Throughput Screening System for Photodynamic Therapy against Cancer Cells
Authors: Rina Lee, Chung-Hun Oh, Eunjin Lee, Jeongyun Kim
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The Photodynamic therapy (PDT) is a treatment that uses a photosensitizer as a drug to damage and kill cancer cells. After injecting the photosensitizer into the bloodstream, the drug is absorbed by cancer cells selectively. Then the area to be treated is exposed to specific wavelengths of light and the photosensitizer produces a form of oxygen that kills nearby cancer cells. PDT is has an advantage to destroy the tumor with minimized side-effects on normal cells. But, PDT is not a completed method for cancer therapy. Because the mechanism of PDT is quite clear yet and the parameters such as intensity of light and dose of photosensitizer are not optimized for different types of cancers. To optimize these parameters, we suggest a novel microfluidic system to automatically control intensity of light exposure with a personal computer (PC). A polydimethylsiloxane (PDMS) microfluidic chip is composed with (1) a cell culture channels layer where cancer cells were trapped to be tested with various dosed photofrin (1μg/ml used for the test) as the photosensitizer and (2) a color dye layer as a neutral density (ND) filter to reduce intensity of light which exposes the cell culture channels filled with cancer cells. Eight different intensity of light (10%, 20%, …, 100%) are generated through various concentrations of blue dye filling the ND filter. As a light source, a light emitting diode (LED) with 635nm wavelength was placed above the developed PDMS microfluidic chip. The total time for light exposure was 30 minutes and HeLa and PC3 cell lines of cancer cells were tested. The cell viability of cells was evaluated with a Live/Dead assay kit (L-3224, Invitrogen, USA). The stronger intensity of light exposed, the lower viability of the cell was observed, and vice versa. Therefore, this system was demonstrated through investigating the PDT against cancer cell to optimize the parameters as critical light intensity and dose of photosensitizer. Our results suggest that the system can be used for optimizing the combinational parameters of light intensity and photosensitizer dose against diverse cancer cell types.Keywords: photodynamic therapy, photofrin, high throughput screening, hela
Procedia PDF Downloads 3833236 Key Roles of the N-Type Oxide Layer in Hybrid Perovskite Solar Cells
Authors: Thierry Pauporté
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Wide bandgap n-type oxide layers (TiO2, SnO2, ZnO etc.) play key roles in perovskite solar cells. They act as electron transport layers, and they permit the charge separation. They are also the substrate for the preparation of perovskite in the direct architecture. Therefore, they have a strong influence on the perovskite loading, its crystallinity and they can induce a degradation phenomenon upon annealing. The interface between the oxide and the perovskite is important, and the quality of this heterointerface must be optimized to limit the recombination of charges phenomena and performance losses. One can also play on the oxide and use two oxide contact layers for improving the device stability and durability. These aspects will be developed and illustrated on the basis of recent results obtained at Chimie-ParisTech.Keywords: oxide, hybrid perovskite, solar cells, impedance
Procedia PDF Downloads 3153235 Investigating Prostaglandin E2 and Intracellular Oxidative Stress Levels in Lipopolysaccharide-Stimulated RAW 264.7 Macrophages upon Treatment with Strobilanthes crispus
Authors: Anna Pick Kiong Ling, Jia May Chin, Rhun Yian Koh, Ying Pei Wong
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Background: Uncontrolled inflammation may cause serious inflammatory diseases if left untreated. Non-steroidal anti-inflammatory drug (NSAIDs) is commonly used to inhibit pro-inflammatory enzymes, thus, reduce inflammation. However, long term administration of NSAIDs leads to various complications. Medicinal plants are getting more attention as it is believed to be more compatible with human body. One of them is a flavonoid-containing medicinal plants, Strobilanthes crispus which has been traditionally claimed to possess anti-inflammatory and antioxidant activities. Nevertheless, its anti-inflammatory activities are yet to be scientifically documented. Objectives: This study aimed to examine the anti-inflammatory activity of S. crispus by investigating its effects on intracellular oxidative stress and prostaglandin E2 (PGE2) levels. Materials and Methods: In this study, the Maximum Non-toxic Dose (MNTD) of methanol extract of both leaves and stems of S. crispus was first determined using 3-(4,5-dimethylthiazolyl-2)-2,5-diphenytetrazolium Bromide (MTT) assay. The effects of S. crispus extracts at MNTD and half MNTD (½MNTD) on intracellular ROS as well as PGE2 levels in 1.0 µg/mL LPS-stimulated RAW 264.7 macrophages were then be measured using DCFH-DA and a competitive enzyme immunoassay kit, respectively. Results: The MNTD of leaf extract was determined as 700µg/mL while for stem was as low as 1.4µg/mL. When LPS-stimulated RAW 264.7 macrophages were subjected to the MNTD of S. crispus leaf extract, both intracellular ROS and PGE2 levels were significantly reduced. In contrast, stem extract at both MNTD and ½MNTD did not significantly reduce the PGE2 level, but significantly increased the intracellular ROS level. Conclusion: The methanol leaf extract of S. crispus may possess anti-inflammatory properties as it is able to significantly reduce the intracellular ROS and PGE2 levels of LPS-stimulated cells. Nevertheless, further studies such as investigating the interleukin, nitric oxide and cytokine tumor necrosis factor-α (TNFα) levels has to be conducted to further confirm the anti-inflammatory properties of S. crispus.Keywords: anti-inflammatory, natural products, prostaglandin E2, reactive oxygen species
Procedia PDF Downloads 2843234 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 1413233 Ovarian Surface Epithelium Receptors during Pregnancy and Estrus Cycle of Rats with Emphasis on Steroids and Gonadotropins Fluctuation
Authors: Salina Yahya Saddik
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The present study is designed to demonstrate the Ovarian Surface Epithelial cells (OSE) Estrogen Receptor α (ERα) and Progesterone Receptor (PR) during pregnancy and estrous cycle in rat. Moreover, determination of the levels of plasma progesterone, estradiol, FSH and LH were also made. The levels of plasma progesterone, estradiol, FSH and LH concentrations were determined on days 7 (n=5), 14 (n=5), and 21(n=5) of pregnancy in three groups of rats and during the estrous cycle (n=5) using ELISA kit. Immunohistochemical method for PR and ERα expression was also made on the ovary. During pregnancy, FSH and LH remained low except at term when LH levels began to increase from 16 ng/ml to 47 ng/ml. Progesterone levels significantly exceeded estradiol values in all pregnant rats with a peak value of 202 ng/ml on day 14. Elevated progesterone levels were associated negatively with LH and estradiol levels during pregnancy. The levels of estradiol surged significantly on day 21. Immunohistochemistry of the ovary showed low levels of OSE cells staining positive for ERα expression. ERα positive cells were absent on day 7 and 14 of pregnancy, only day 21 recorded a very low percentage of immunostaining (0.5%) within the nuclei of OSE cells. On the contrary, immunostaining of PR was not observed within the nuclei of OSE cells in all groups of study. In conclusions, these results may suggest that progesterone effect during pregnancy seems to be overriding the positive effect of estrogens on OSE cells. High progesterone levels may have a direct negative effect on gonadotropin production and thereby it might inhibit events leading to both follicular development and OSE proliferation. Understanding the factors affecting OSE proliferation may help elucidating the mechanism(s) of assisted diseases such as ovarian cancer.Keywords: ovarian surface, pregnancy, gonadotropins, steroids
Procedia PDF Downloads 3143232 The Role of a Biphasic Implant Based on a Bioactive Silk Fibroin for Osteochondral Tissue Regeneration
Authors: Lizeth Fuentes-Mera, Vanessa Perez-Silos, Nidia K. Moncada-Saucedo, Alejandro Garcia-Ruiz, Alberto Camacho, Jorge Lara-Arias, Ivan Marino-Martinez, Victor Romero-Diaz, Adolfo Soto-Dominguez, Humberto Rodriguez-Rocha, Hang Lin, Victor Pena-Martinez
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Biphasic scaffolds in cartilage tissue engineering have been designed to influence not only the recapitulation of the osteochondral architecture but also to take advantage of the healing ability of bone to promote the implant integration with the surrounding tissue and then bone restoration and cartilage regeneration. This study reports the development and characterization of a biphasic scaffold based on the assembly of a cartilage phase constituted by fibroin biofunctionalized with bovine cartilage matrix; cellularized with differentiated pre-chondrocytes from adipose tissue stem cells (autologous) and well attached to a bone phase (bone bovine decellularized) to mimic the structure of the nature of native tissue and to promote the cartilage regeneration in a model of joint damage in pigs. Biphasic scaffolds were assembled by fibroin crystallization with methanol. The histological and ultrastructural architectures were evaluated by optical and scanning electron microscopy respectively. Mechanical tests were conducted to evaluate Young's modulus of the implant. For the biological evaluation, pre-chondrocytes were loaded onto the scaffolds and cellular adhesion, proliferation, and gene expression analysis of cartilage extracellular matrix components was performed. The scaffolds that were cellularized and matured for 10 days were implanted into critical 3 mm in diameter and 9-mm in depth osteochondral defects in a porcine model (n=4). Three treatments were applied per knee: Group 1: monophasic cellular scaffold (MS) (single chondral phase), group 2: biphasic scaffold, cellularized only in the chondral phase (BS1), group 3: BS cellularized in both bone and chondral phases (BS2). Simultaneously, a control without treatment was evaluated. After 4 weeks of surgery, integration and regeneration tissues were analyzed by x-rays, histology and immunohistochemistry evaluation. The mechanical assessment showed that the acellular biphasic composites exhibited Young's modulus of 805.01 kPa similar to native cartilage (400-800 kPa). In vitro biological studies revealed the chondroinductive ability of the biphasic implant, evidenced by an increase in sulfated glycosaminoglycan (GAGs) and type II collagen, both secreted by the chondrocytes cultured on the scaffold during 28 days. No evidence of adverse or inflammatory reactions was observed in the in vivo trial; however, In group 1, the defects were not reconstructed. In group 2 and 3 a good integration of the implant with the surrounding tissue was observed. Defects in group 2 were fulfilled by hyaline cartilage and normal bone. Group 3 defects showed fibrous repair tissue. In conclusion; our findings demonstrated the efficacy of biphasic and bioactive scaffold based on silk fibroin, which entwined chondroinductive features and biomechanical capability with appropriate integration with the surrounding tissue, representing a promising alternative for osteochondral tissue-engineering applications.Keywords: biphasic scaffold, extracellular cartilage matrix, silk fibroin, osteochondral tissue engineering
Procedia PDF Downloads 153