Search results for: CD8+ T cell responses

Authors: Inna Kornienko, Svetlana Guryeva, Natalia Danilova, Elena Petersen

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Drug toxicity often goes undetected until clinical trials, the most expensive and dangerous phase of drug development. Both human cell culture and animal studies have limitations that cannot be overcome by improvements in drug testing protocols. Tissue engineering is an emerging alternative approach to creating models of human malignant tumors for experimental oncology, personalized medicine, and drug discovery studies. This new generation of bioengineered tumors provides an opportunity to control and explore the role of every component of the model system including cell populations, supportive scaffolds, and signaling molecules. An area that could greatly benefit from these models is cancer research. Recent advances in tissue engineering demonstrated that decellularized tissue is an excellent scaffold for tissue engineering. Decellularization of donor organs such as heart, liver, and lung can provide an acellular, naturally occurring three-dimensional biologic scaffold material that can then be seeded with selected cell populations. Preliminary studies in animal models have provided encouraging results for the proof of concept. Decellularized Organs preserve organ microenvironment, which is critical for cancer metastasis. Utilizing 3D tumor models results greater proximity of cell culture morphological characteristics in a model to its in vivo counterpart, allows more accurate simulation of the processes within a functioning tumor and its pathogenesis. 3D models allow study of migration processes and cell proliferation with higher reliability as well. Moreover, cancer cells in a 3D model bear closer resemblance to living conditions in terms of gene expression, cell surface receptor expression, and signaling. 2D cell monolayers do not provide the geometrical and mechanical cues of tissues in vivo and are, therefore, not suitable to accurately predict the responses of living organisms. 3D models can provide several levels of complexity from simple monocultures of cancer cell lines in liquid environment comprised of oxygen and nutrient gradients and cell-cell interaction to more advanced models, which include co-culturing with other cell types, such as endothelial and immune cells. Following this reasoning, spheroids cultivated from one or multiple patient-derived cell lines can be utilized to seed the matrix rather than monolayer cells. This approach furthers the progress towards personalized medicine. As an initial step to create a new ex vivo tissue engineered model of a cancer tumor, optimized protocols have been designed to obtain organ-specific acellular matrices and evaluate their potential as tissue engineered scaffolds for cultures of normal and tumor cells. Decellularized biomatrix was prepared from animals’ kidneys, urethra, lungs, heart, and liver by two decellularization methods: perfusion in a bioreactor system and immersion-agitation on an orbital shaker with the use of various detergents (SDS, Triton X-100) in different concentrations and freezing. Acellular scaffolds and tissue engineered constructs have been characterized and compared using morphological methods. Models using decellularized matrix have certain advantages, such as maintaining native extracellular matrix properties and biomimetic microenvironment for cancer cells; compatibility with multiple cell types for cell culture and drug screening; utilization to culture patient-derived cells in vitro to evaluate different anticancer therapeutics for developing personalized medicines.

Keywords: 3D models, decellularization, drug discovery, drug toxicity, scaffolds, spheroids, tissue engineering

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Authors: Ganesh K. Maurya, Reema Chaudhary, Neha Pandey, Hari S. Misra

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PprA, a pleiotropic protein participating in radioresistance, has been reported for its roles in DNA replication initiation, genome segregation, cell division and DNA repair in polyextremophile Deinococcus radiodurans. Interestingly, expression of deinococcal PprA in E. coli suppresses its growth by reducing the number of colony forming units and provide better resistance against γ-radiation than control. We employed different biochemical and cell biology studies using PprA and its DNA binding/polymerization mutants (K133E & W183R) in E. coli. Cells expressing wild type PprA or its K133E mutant showed reduction in the amount of genomic DNA as well as chromosome copy number in comparison to W183R mutant of PprA and control cells, which suggests the role of PprA protein in regulation of DNA replication initiation in E. coli. Further, E. coli cells expressing PprA or its mutants exhibited different impact on cell morphology than control. Expression of PprA or K133E mutant displayed a significant increase in cell length upto 5 folds while W183R mutant showed cell length similar to uninduced control cells. We checked the interaction of deinococcal PprA and its mutants with E. coli DnaA using Bacterial two-hybrid system and co-immunoprecipitation. We observed a functional interaction of EcDnaA with PprA and K133E mutant but not with W183R mutant of PprA. Further, PprA or K133E mutant has suppressed the ATPase activity of EcDnaA but W183R mutant of PprA failed to do so. These observations suggested that PprA protein regulates DNA replication initiation and cell morphology of surrogate E. coli.

Keywords: DNA replication, radioresistance, protein-protein interaction, cell morphology, ATPase activity

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Authors: Singh Deepeshwar, Suhas Vinchurkar

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Similar to neuroimaging findings through functional magnetic resonance imaging (fMRI) assessing regional cerebral blood oxygenation, the functional near infrared spectroscopy (fNIRS) has also been used to assess hemodynamic responses in the imaged region of the brain. The present study assessed hemodynamic responses in terms of changes in oxygenation (HbO), deoxygenation (HbR) and total hemoglobin (THb) on the prefrontal cortex (PFC), bilaterally, using fNIRS in 10 participants who performed three voluntary regulated breathing (pranayama) practices viz. (i) Left nostril breathing (LNB), (ii) Right nostril breathing (RNB); and (iii) Alternating nostril breathing (ANB) and compared with normal breathing as baseline (BS). For this, we used 64 channel NIRS system covering left and the right prefrontal cortex. The normal breathing kept as baseline (BS) measures as regressors in the investigation of hemodynamic responses when compared with LNB, RNB and ANB. In the results, we found greater oxygenation in contralateral side i.e., higher activation on the left prefrontal cortex (lPFC) during RNB, and right prefrontal cortex (rPFC) during LNB, whereas ANB showed greater deoxygenation responses on both sides of PFC. Interestingly, LNB showed increased oxygenation on ipsilateral side i.e., lPFC but not during RNB. This suggests that voluntary regulated breathing produced an immediate effect not only on contralateral but ipsilateral sides of the brain as well. In conclusion, breathing practices are tightly coupled to cerebral rhythms of alternating cerebral hemispheric activity during particular nostril breathing. These results of the specific nostril breathing do not support previous findings of contralateral hemispheric improvement while left or right nostril breathing only.

Keywords: hemodynamic responses, brain, pranayama, voluntary regulated breathing practices, prefrontal cortex

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Authors: Roshika Tyagi, Shuvomoy Banerjee

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Among various diseases, cancer has become a huge threat to human beings globally. In the context of viral infection, Epstein–Barr virus (EBV) infection is ubiquitous in nature world-wide as well as in India. Recepteur d’Origine Nantais (RON) receptor tyrosine kinase is overexpressed in Lymphoblastoid cell lines (LCLs) but undetectable in primary B-cells. Biologically, RON expression was found to be essential for EBV transformed LCLs proliferation. In our study, we investigated whether EBV latent antigen EBNA3C is playing a crucial role in regulating RON receptor tyrosine kinase function in EBV-induced malignancies. Interestingly, we observed that expression pattern of RON was modulated by EBNA3C in EBV transformed LCLs compared with EBV negative BJAB cell line by PCR and western blot analysis. Moreover, in the absence of EBNA3C, RON expression was found low in western blot and immunofluorescence analysis and cell proliferation rate was significantly reduced in LCLs by cell viability assays. Therefore, our study clearly indicating the potential role of EBNA3C expressed in EBV-infected B-cells for modulating the functions of oncogenic kinases that leads to EBV induced B-cell transformation.

Keywords: apoptosis, cell proliferation, Epstein–barr virus, receptor tyrosine kinase

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Authors: Ediati Budi Cahyono, Triana Hertiani, Nauval Arrazy Asawimanda, Wahyu Puji Pratomo

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Background: Doxorubicin is widely used as a chemotherapeutic drug despite having many side effects. It may cause macrophage dysfunction and decreasing proliferation of lymphocyte. Noni (Morinda citrifolia) fruit which has rich of polysaccharide content has potential as antitumor and immunostimulant effect. The isolation of polysaccharide from Noni fruit has been optimized according to four different methods based on macrophage and lymphocyte activities. We found the highest polysaccharide content from one of the four methods isolation. A method of polysaccharide isolation which has the highest immunostimulant effect was used for further observation as co-chemotherapy. The aim of the study: was to evaluate the isolated polysaccharide from the method of choice as co-chemotherapy of doxorubicin for the growth of Vero, He-La, and T47D cell lines in vitro. The method: in vitro growth assay of Vero, He-La, and T47D cell lines was done using MTT-reduction method, and apoptosis test was done by double staining method to evaluate the induction apoptotic effect of the combination. Every group was treated with doxorubicin and isolated polysaccharide from method of choice with 4 variances of concentrations (25 µg/ml, 50 µg/ml, 100 µg/ml and 200 µg/ml) a long with negative control (doxorubicin only) and normal control (without doxorubicin or polysaccharide administration). Results: The combination of polysaccharide fraction in the concentration of 100μg/ml with 2μmol of doxorubicin against He-La and T47D cell lines influenced the highest cytotoxic effect by suppressing cell viability comparing with doxorubicin only. The combination of polysaccharide fraction in the concentration of 100μg/ml with 2μmol of doxorubicin-induced apoptotic effect the He-La cell line comparing with doxorubicin only. The result of the study: it can be concluded that the combination of polysaccharide fraction and doxorubicin effect more selective toward He-La and T47D cell lines than to Vero cell line. It can be suggested isolated polysaccharide from the method of choice has co-chemotherapy activity against doxorubicin.

Keywords: polysaccharide, noni fruit, doxorubicin, cancer cell lines, vero cell line

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Authors: Rawan Al-Nemari, Abdulrahman Al-Senaidy, Abdelhabib Semlali

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Background and objectives: The most common cause of death in women worldwide is breast cancer. Regarding all chemotherapy disadvantages and side effects, it’s becoming necessary to identify natural products that target cancer cells with lesser harmful side effects on non-targeted cells and biological environment. Different parts of A. squamosa L., commonly known as custard apple, show varied therapeutic effects. The objective of this study is to investigate in vitro and in vivo, the anti-cancer activity of A. squamosa leaves extract. Methods: The physiological responses using MTT, nucleus staining, and LDH assays were used to evaluate cell survival and proliferation in both ER+ and ER- cells when they were exposed to extract. Monolayer wound repair assay was used to investigate the effect of extracts on cell migration. Apoptotic gene’s expression was investigated by real-time polymerase chain reaction. To study the effect of the extract on the size of tumor, breast cancer induced rats were used. Results: A. squamosa leaves extract showed high anti-proliferative and cytotoxicity effects against different breast cancer cell lines with high concentration, 100 ug/ml. The extracts have reduced the cells wound closure. Polymerase chain reaction revealed downregulation of Bcl-2 and upregulation of Bax. In breast cancer model animal developed in our laboratory, after 4 weeks treatment, treated groups have shown smaller tumor size in comparison with control group (n=4). Conclusion: These results suggest that A. squamosa leaves extract has anti-cancer activity against breast cancer in both in vitro and in vivo, and it may be developed as a potential novel agent to treat breast cancer.

Keywords: apoptosis, breast cancer, migration, proliferation

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Authors: A. O. Boyo, A. T. Akinwunmi

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The effect of extracting solvent and adjustment of pHs on the stability of Terminalia catappa L. dye-sensitized solar cell was investigated. We introduced ZnO as an alternative to TiO2 in the dye sensitized solar cells (DSSCs) due to its band gap similar to TiO2, higher electron mobility, and flexible procedures of preparations. Dye-sensitized solar cells (DSSCs) based on Terminalia catappa L. was extracted in water (A), ethanol (B) and the mixture of ethanol and water in the ratio 1:1by volume (C). The best performance Solar cells sensitized was from extracts A and achieved up to Jsc 1.51 mAcm−2, Voc 0.75V, FF 0.88 and η 0.63%. We notice that as pHs decreases there is the increase in DSSC efficiency. There is Long period stability in efficiency of the cells prepared using A than in C and a fair stability in efficiency of B cell. The results obtained with extracts B and C confirmed that Ethanol with water could not be considered as a suitable solvent for the extraction of natural dye.

Keywords: zinc oxide, dye-sensitized solar cell, terminalia catappa L., TiO2

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Authors: M. S. Kilic, S. Korkut, B. Hazer

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Newly synthesized Polypropylene-g-Polyethylene glycol polymer was first time used for a compartment-less enzymatic fuel cell. Working electrodes based on Polypropylene-g-Polyethylene glycol were operated as unmediated and mediated system (with ferrocene and gold/cobalt oxide nanoparticles). Glucose oxidase and bilirubin oxidase was selected as anodic and cathodic enzyme, respectively. Glucose was used as fuel in a single-compartment and membrane-less cell. Maximum power density was obtained as 0.65 nW cm-2, 65 nW cm-2, and 23500 nW cm-2 from the unmediated, ferrocene and gold/cobalt oxide modified polymeric film, respectively. Power density was calculated to be ~16000 nW cm-2 for undiluted wastewater sample with gold/cobalt oxide nanoparticles including system.

Keywords: bilirubin oxidase, enzymatic fuel cell, glucose oxidase, nanoparticles

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Authors: Jayprakash Yadav, Nivedita Patra

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Polyhydroxybutyrate (PHB) is an interesting material in the field of medical science, pharmaceutical industries, and tissue engineering because of its properties such as biodegradability, biocompatibility, hydrophobicity, and elasticity. PHB is naturally accumulated by several microbes in their cytoplasm during the metabolic process as energy reserve material. PHB can be extracted from cell biomass using halogenated hydrocarbons, chemicals, and enzymes. In this study, a cheaper and non-toxic solvent, acetone, was used for the extraction process. The different parameters like acetone percentage, and solvent pH, process temperature, and incubation periods were optimized using the Response Surface Methodology (RSM). RSM was performed and the determination coefficient (R2) value was found to be 0.8833 from the quadratic regression model with no significant lack of fit. The designed RSM model results indicated that the fitness of the response variable was significant (P-value < 0.0006) and satisfactory to denote the relationship between the responses in terms of PHB recovery and purity with respect to the values of independent variables. Optimum conditions for the maximum PHB recovery and purity were found to be solvent pH 7, extraction temperature - 43 °C, incubation time - 70 minutes, and percentage acetone – 30 % from this study. The maximum predicted PHB recovery was found to be 0.845 g/g biomass dry cell weight and the purity was found to be 97.23 % using the optimized conditions.

Keywords: acetone, PHB, RSM, halogenated hydrocarbons, extraction, bacillus subtilis.

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Authors: Ainur Mukhambetova, Miras Karzhauov, Vyacheslav Ogay

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Introduction: Mesenchymal stem cells (MSCs) have the capacity to be differentiated into several cell lineages and are a promising source for cell therapy and tissue engineering. However, currently most MSCs culturing protocols use media supplemented with fetal bovine serum (FBS), which limits their application in clinic due to the possibility of zoonotic infections, contamination and immunological reactions. Consequently, formulating effective serum free culture medium becomes one of the important problems in contemporary cell biotechnology. Objectives: The aim of this study was to define an optimal serum-free medium for culturing of periosteum derived MSCs. Materials and methods: The MSCs were extracted from human periosteum and transferred to the culture flasks pretreated with CELLstart™. Immunophenotypic characterization, proliferation and in vitro differentiation of cells grown on STEM PRO® MSC SFM were compared to the cells cultured in the standard FBS containing media. Chromosome analysis and flow cytometry were also performed. Results: We have shown that cells were grown on STEM PRO® MSC SFM retained all the morphological, immunophenotypic (CD73, CD90, CD105, vimentin and Stro-1) and cell differentiation characteristics specific to MSCs. Chromosome analysis indicated no anomalies in the chromosome structure. Flow cytometry showed a high expression of cell adhesion molecules CD44 (98,8%), CD90 (97,4%), CD105 (99,1%). In addition, we have shown that cell is grown on STEM PRO® MSC SFM have higher proliferation capacity compared to cell expanded on standard FBS containing the medium. Conclusion: We have shown that STEM PRO® MSC SFM is optimal for culturing periosteum derived human MSCs which subsequently can be safely used in cell therapy.

Keywords: cell technologies, periosteum-derived MSCs, regenerative medicine, serum-free medium

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Authors: Nabila N. El-Maraghy, Amany Essa, Yousra Abdel–Mottaleb, Nada Ismail

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The use of combination of chemotherapy and natural products to influence the cell death with low doses of chemotherapeutic agents and few side effects has recently emerged as a new method of cancer therapy. Aim: Evaluation the modulatory effect of Thymoquinone on HepG2 cells treated with Sorafenib. Methods: Hepatocellular Carcinoma HepG2 cell line was treated with Sorafenib and TQ individually and in combination. The effect of these treatments on cell viability (MTT assay), apoptosis (Expression of Caspase-3) and oxidative markers (GSH content and extent of lipid peroxidation) was determined. Results: When compared the effect of both agents alone and the combination of the IC50 of Sorafenib and the IC50 TQ, the combination resulted in reduction of cell inhibition and apoptosis and antagonize their actions on GSH content and extent of lipid peroxidation which are increased. This study showed potent anti-tumor activity of both TQ and Sorafenib separately on HepG2 but upon combination surprisingly they interacted and give antagonistic effect. Conclusion: Co-treatment resulted in antagonistic interaction between Sorafenib and Thymoquinone.

Keywords: antagonism, hepatocellular carcinoma, sorafenib, thymoquinone

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Authors: Shih-cheng Li, Ming-Yi Chung, Mei-Lien Chen

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Background: Plasticizers, such as phthalates and adipates, were substances added to a material that provided flexibility and durability to plastics such as polyvinyl chloride (PVC). Phthalates were generally recognized as an endocrine disrupter due to their estrogenic and anti-androgenic activities. Phthalates had the capacity to bind to estrogen receptors, and hence they might prolong menstrual cycles and increase the proportion of premature menopause. Recently, adipates such as di-2-ethylhexyl adipate (DEHA) and di-isononyl adipate (DiNA) had replaced phthalates and were now used for food packaging. Methods: MCF-7 cell lines were treated with di-2-ethylhexyl phthalate (DEHP), di- 2-ethylhexyl adipate (DEHA), or di-isononyl adipate (DiNA) (10-6 , 10-5 , and 10-4 mol/l), using 17β-estradiol (10-8 mol/l) as a positive control. After incubations of 24, 48, 72, and 96 hours, the cells were tested using the alamarBlue assay. Results: The alamarBlue assay revealed that cell proliferation significantly increased after treatments of DEHP and DEHA for 24 hours at a concentration of 10-6, 10-5, and 10-4 mol/l. After more than 48 hours, cell proliferations in DEHP at 10-6, 10-5, and 10-4 mol/l significantly decreased compared to the control group. Conclusions: The present study demonstrates that adipates, as well as phthalates, were capable of inducing cell proliferation. We further used MDA-MB-231 cell lines to confirm that the proliferation effect was generated through binding to estrogen receptors.

Keywords: MCF-7, phthalate, adipate, endocrine disrupter

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Authors: Quan Gu, Daniel Dimitrov

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BingleSeq was developed as a shiny-based, intuitive, and comprehensive application that enables the analysis of single-Cell RNA-Sequencing count data. This was achieved via incorporating three state-of-the-art software packages for each type of RNA sequencing analysis, alongside functional annotation analysis and a way to assess the overlap of differential expression method results. At its current state, the functionality implemented within BingleSeq is comparable to that of other applications, also developed with the purpose of lowering the entry requirements to RNA Sequencing analyses. BingleSeq is available on GitHub and will be submitted to R/Bioconductor.

Keywords: bioinformatics, functional annotation analysis, single-cell RNA-sequencing, transcriptomics

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Authors: Syed Sadique Anwer Askari, Mukul Kumar Das

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Zinc Oxide (ZnO) solar cells have drawn great attention due to the enhanced efficiency and low-cost fabrication process. In this study, ZnO thin film is used as the active layer, hole blocking layer, antireflection coating (ARC) as well as transparent conductive oxide. To improve the conductivity of ZnO, top layer of ZnO is doped with aluminum, for top contact. Intrinsic carrier concentration of silicon substrate plays an important role in enhancing the power conversion efficiency (PCE) of ZnO/Si solar cell. With the increase of intrinsic carrier concentration PCE decreased due to increase in dark current in solar cell. At 80nm ZnO and 160µm Silicon substrate thickness, power conversion efficiency of 26.45% and 21.64% is achieved with intrinsic carrier concentration of 1x109/cm3, 1.4x1010/cm3 respectively.

Keywords: hetero-junction solar cell, solar cell, substrate intrinsic carrier concentration, ZnO/Si

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Authors: Agatha Bebbington, Terry Tetley, Kathryn Hadler

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Background: Astronauts will set foot on the Moon later this decade, and are at high risk of lunar dust inhalation. Freshly-fractured lunar dust produces reactive oxygen species in solution, which are known to cause cellular damage and inflammation. Cytotoxicity and inflammatory mediator release was measured in pulmonary alveolar epithelial cells (cells that line the gas-exchange zone of the lung) exposed to a lunar dust simulant, LMS-1. It was hypothesised that freshly-fractured LMS-1 would result in increased cytotoxicity and inflammatory mediator release, owing to the angular morphology and high reactivity of fractured particles. Methods: A human alveolar epithelial type 1-like cell line (TT1) and a human macrophage-like cell line (THP-1) were exposed to 0-200μg/ml of unground, aged-ground, and freshly-ground LMS-1 (screened at <22μm). Cell viability, cytotoxicity, and inflammatory mediator release (IL-6, IL-8) were assessed using MMT, LDH, and ELISA assays, respectively. LMS-1 particles were characterised for their size, surface area, and morphology before and after grinding. Results: Exposure to LMS-1 particles did not result in overt cytotoxicity in either TT1 epithelial cells or THP-1 macrophage-like cells. A dose-dependent increase in IL-8 release was observed in TT1 cells, whereas THP-1 cell exposure, even at low particle concentrations, resulted in increased IL-8 release. Both cytotoxic and pro-inflammatory responses were most marked and significantly greater in TT1 and THP-1 cells exposed to freshly-fractured LMS-1. Discussion: LMS-1 is a novel lunar dust simulant; this is the first study to determine its toxicological effects on respiratory cells in vitro. An increased inflammatory response in TT1 and THP-1 cells exposed to ground LMS-1 suggests that low particle size, increased surface area, and angularity likely contribute to toxicity. Conclusions: Evenlow levels of exposure to LMS-1 could result in alveolar inflammation. This may have pathological consequences for astronauts exposed to lunar dust on future long-duration missions. Future research should test the effect of low-dose, intermittent lunar dust exposure on the respiratory system.

Keywords: lunar dust, LMS-1, lunar dust simulant, long-duration space travel, lunar dust toxicity

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Authors: H. Katkhuda

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A time domain approach is used in this paper to identify unknown dynamic forces applied on two dimensional frames using the measured dynamic structural responses for a sub-structure in the two dimensional frame. In this paper a sub-structure finite element model with short length of measurement from only three or four accelerometers is required, and an iterative least-square algorithm is used to identify the unknown dynamic force applied on the structure. Validity of the method is demonstrated with numerical examples using noise-free and noise-contaminated structural responses. Both harmonic and impulsive forces are studied. The results show that the proposed approach can identify unknown dynamic forces within very limited iterations with high accuracy and shows its robustness even noise- polluted dynamic response measurements are utilized.

Keywords: dynamic force identification, dynamic responses, sub-structure, time domain

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Authors: Karen K. Perez-Ramirez, Genevieve Dupont, Virginia Gonzalez-Velez

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Pancreatic alpha-cells participate on glucose regulation together with beta cells. They release glucagon hormone when glucose level is low to stimulate gluconeogenesis from the liver. As other excitable cells, alpha cells generate Ca2+ and metabolic oscillations when they are stimulated. It is known that the glucose level can trigger or silence this activity although it is not clear how this occurs in normal and diabetic people. In this work, we propose an electric-metabolic mathematical model implemented in Matlab to study the effect of different glucose levels on the electrical response and Ca2+ oscillations of an alpha cell. Our results show that Ca2+ oscillations appear in opposite phase with metabolic oscillations in a window of glucose values. The model also predicts a direct relationship between the level of glucose and the intracellular adenine nucleotides showing a self-regulating pathway for the alpha cell.

Keywords: Ca2+ oscillations, mathematical model, metabolic oscillations, pancreatic alpha cell

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Authors: Engin Berber, Nurettin Canakoglu, Ibrahim Sozdutmaz, Merve Caliskan, Shaikh Terkis Islam Pavel, Hazel Yetiskin, Aykut Ozdarendeli

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Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne virus in the family Bunyaviridae, genus Nairovirus. The CCHFV genome consists of three molecules of negative-sense single-stranded RNA, each encapsulated separately. The virion particle contains viral RNA polymerase (L segment), surface glycoproteins Gn and Gc (Msegment), and a nucleocapsid protein NP (S segment). CCHF is characterized by high case mortality, occurring in Asia, Africa, the Middle East and Eastern Europe. Clinical CCHF was first recognized in Turkey in 2002. The numbers of CCHF cases have gradually increased in Turkey making the virus a public health concern. Between 2002 and 2014, more than 8000 the CCHF cases have been reported in Turkey and mortality rate is around 5%. So, Turkey is one of the countries where the epidemy has become spread to the wider geography and the biggest outbreaks of CCHF have occurred in the world. We have recently developed an inactivated cell-culture based vaccine against CCHF. We have showed that the Balb/c mice immunized with the CCHF vaccine induced the high level of neutralizing antibodies. In this study, we aimed to determine the immunodominant regions of nucleoprotein (NP) CCHFV Kelkit06 strain which stimulate T cells. For this purpose, pools of overlapping NP were used for an IFN- γ ELISPOT assay. Balb/c mice were divided into two groups for the experiment. Two groups (n = 10 each) were immunized via the intraperitoneal route with 5, or 10μg of the cell culture-based vaccine. The control group (n = 6) was mock immunized with PBS. Booster injections with the same formulation were given on days 21 and 42 after the first immunization. The higher reactivity against the CCHFV NP pools 31-40 and 80-90 was determined in the two dose groups. In order to analyze the vaccine-induced T cell responses in Balb/c mice immunized with varying doses of the vaccine, we have been also currently working on CD4+, CD8+ and CD3 + T cells by flow cytometry.

Keywords: Crimean-Congo hemorrhagic fever virus, immunodominant regions of NP, T cell response, vaccine

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Authors: C. Yin, B. Zhang, Y. Liu, J. Cai

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Airborne EM (AEM) is an effective geophysical exploration tool, especially suitable for ridged mountain areas. In these areas, topography will have serious effects on AEM system responses. However, until now little study has been reported on topographic effect on airborne EM systems. In this paper, an edge-based unstructured finite-element (FE) method is developed for 3D topographic modeling for both frequency and time-domain airborne EM systems. Starting from the frequency-domain Maxwell equations, a vector Helmholtz equation is derived to obtain a stable and accurate solution. Considering that the AEM transmitter and receiver are both located in the air, the scattered field method is used in our modeling. The Galerkin method is applied to discretize the Helmholtz equation for the final FE equations. Solving the FE equations, the frequency-domain AEM responses are obtained. To accelerate the calculation speed, the response of source in free-space is used as the primary field and the PARDISO direct solver is used to deal with the problem with multiple transmitting sources. After calculating the frequency-domain AEM responses, a Hankel’s transform is applied to obtain the time-domain AEM responses. To check the accuracy of present algorithm and to analyze the characteristic of topographic effect on airborne EM systems, both the frequency- and time-domain AEM responses for 3 model groups are simulated: 1) a flat half-space model that has a semi-analytical solution of EM response; 2) a valley or hill earth model; 3) a valley or hill earth with an abnormal body embedded. Numerical experiments show that close to the node points of the topography, AEM responses demonstrate sharp changes. Special attentions need to be paid to the topographic effects when interpreting AEM survey data over rugged topographic areas. Besides, the profile of the AEM responses presents a mirror relation with the topographic earth surface. In comparison to the topographic effect that mainly occurs at the high-frequency end and early time channels, the EM responses of underground conductors mainly occur at low frequencies and later time channels. For the signal of the same time channel, the dB/dt field reflects the change of conductivity better than the B-field. The research of this paper will serve airborne EM in the identification and correction of the topographic effects.

Keywords: 3D, Airborne EM, forward modeling, topographic effect

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Authors: Amin Alvani

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Multiple sclerosis (MS) is a chronic, often disabling disease in which the immune system attacks the myelin sheath of neurons in the central nervous system. The present study aimed to investigate the Relationship between behavioral inhibition/approach system (BIS-BAS) and perceived stress (PS) whit control white blood cell (WBC). 60 MS patients (male=36.7, female=63.3%; age range=15-65 participated in the study and completed the demographic questionnaire, the count blood cell (CBC) test, the behavioral Activation and behavioral inhibition scale (BIS-BAS), and the perceived stress Questionnaire (PSS-14). The results revealed that Between of BAS-reward responsiveness (BAS-DR) subscale and PS, in more than MS patient (BIS), there are increase WBC.

Keywords: behavioral inhibition/approach system, perceived stress, white blood cell, multiple sclerosis

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Authors: Elsie M. Nolte, Annie M. Joubert, Roy Lakier, Maryke Etsebeth, Jolene M. Helena, Marcel Verwey, Laurence Lafanechere, Anne E. Theron

Abstract:

Treatment regimens for breast- and lung cancers may include both radiation- and chemotherapy. Ideally, a pharmaceutical agent which selectively sensitizes cancer cells to gamma (γ)-radiation would allow administration of lower doses of each modality, yielding synergistic anti-cancer benefits and lower metastasis occurrence, in addition to decreasing the side-effect profiles. A range of 2-methoxyestradiol (2-ME) analogues, namely 2-ethyl-3-O-sulphamoyl-estra-1,3,5 (10) 15-tetraene-3-ol-17one (ESE-15-one), 2-ethyl-3-O-sulphamoyl-estra-1,3,5(10),15-tetraen-17-ol (ESE-15-ol) and 2-ethyl-3-O-sulphamoyl-estra-1,3,5(10)16-tetraene (ESE-16) were in silico-designed by our laboratory, with the aim of improving the parent compound’s bioavailability in vivo. The main effect of these compounds is the disruption of microtubule dynamics with a resultant mitotic accumulation and induction of programmed cell death in various cancer cell lines. This in vitro study aimed to determine the cellular responses involved in the radiation sensitization effects of these analogues at low doses in breast- and lung cancer cell lines. The oestrogen receptor positive MCF-7-, oestrogen receptor negative MDA-MB-231- and triple negative BT-20 breast cancer cell lines as well as the A549 lung cancer cell line were used. The minimal compound- and radiation doses able to induce apoptosis were determined using annexin-V and cell cycle progression markers. These doses (cell line dependent) were used to pre-sensitize the cancer cells 24 hours prior to 6 gray (Gy) radiation. Experiments were conducted on samples exposed to the individual- as well as the combination treatment conditions in order to determine whether the combination treatment yielded an additive cell death response. Morphological studies included light-, fluorescence- and transmission electron microscopy. Apoptosis induction was determined by flow cytometry employing annexin V, cell cycle analysis, B-cell lymphoma 2 (Bcl-2) signalling, as well as reactive oxygen species (ROS) production. Clonogenic studies were performed by allowing colony formation for 10 days post radiation. Deoxyribonucleic acid (DNA) damage was quantified via γ-H2AX foci and micronuclei quantification. Amplification of the p53 signalling pathway was determined by western blot. Results indicated that exposing breast- and lung cancer cells to nanomolar concentrations of these analogues 24 hours prior to γ-radiation induced more cell death than the compound- and radiation treatments alone. Hypercondensed chromatin, decreased cell density, a damaged cytoskeleton and an increase in apoptotic body formation were observed in cells exposed to the combination treatment condition. An increased number of cells present in the sub-G1 phase as well as increased annexin-V staining, elevation of ROS formation and decreased Bcl-2 signalling confirmed the additive effect of the combination treatment. In addition, colony formation decreased significantly. p53 signalling pathways were significantly amplified in cells exposed to the analogues 24 hours prior to radiation, as was the amount of DNA damage. In conclusion, our results indicated that pre-treatment of breast- and lung cancer cells with low doses of 2-ME analogues sensitized breast- and lung cancer cells to γ-radiation and induced apoptosis more so than the individual treatments alone. Future studies will focus on the effect of the combination treatment on non-malignant cellular counterparts.

Keywords: cancer, microtubule dynamics, radiation therapy, radiosensitization

Procedia PDF Downloads 179

Authors: Z. Mahmood, F. U. Babar, S. Naz, H. U. Rehman

Abstract:

Cadmium Sulphide (CdS) was deposited on a Tec 15 glass substrate with the help of CBD (chemical bath deposition process) and then cadmium telluride CdTe was deposited on CdS with the help of CSS (closed spaced sublimation technique) for the construction of a solar cell. The thicknesses of all the deposited materials were measured with the help of Ellipsometry. The IV graphs were drawn in order to observe the current voltage output. The efficiency of the cell was graphed with the fill factor as well (graphs not given here). The efficiency came out to be approximately 16.5 % and the CIGS (copper-indium–gallium-selenide) maximum efficiency is 20 %. The efficiency of a solar cell can further be enhanced by adapting quality materials, good experimental devices and proper procedures. The grain size was analyzed with the help of scanning electron microscope using RBS (Rutherford backscattering spectroscopy).

Keywords: Chemical Bath Deposition Technique (CBD), cadmium sulphide (CdS), CdTe, CSS (Closed Space Sublimation)

Procedia PDF Downloads 327

Authors: Seul Ki Min, Gwang Heum Yoon, Jung Hyun Joo, Hwa Sung Shin

Abstract:

Cellular membrane distortion is known as a factor to change intracellular signaling. However, progress of relevant studies is difficult because there are no facilities that can control membrane distortion finely. In this study, we developed microfluidic device which can inflict mechanical stress on cell membrane of Chlamydomonas reinhardtii using regular height of the channels. And cellular physiological changes were analyzed from cells cultured in the device. Excessive calcium ion influx through into cytoplasm was induced from mechanical stress. The results revealed that compressed cells had up-regulated Mat3 mRNA which regulates cell size and cell cycle from a prolonged G1 phase. Additionally, TAG used for the production of biodiesel was raised rapidly from 4 h after compression. Taken together, membrane distortion can be considered as an attractive inducer for biofuel production.

Keywords: mechanical stress, membrane distortion, Chlamydomonas reinhardtii, deflagellation, cell cycle, lipid metabolism

Procedia PDF Downloads 341

Authors: Ji Un Shin, Wei Mao, Hyuk Sang Yoo

Abstract:

Electrospinning is a versatile tool for fabricating nano-structured polymeric materials. Gelatin hydrogels are considered to be a good material for cell cultivation because of high water swellability as well as good biocompatibility. Three-dimensional (3-D) cell cultivation is a desirable method of cell cultivation for preparing tissues and organs because cell-to-cell interactions or cell-to-matrix interactions can be much enhanced through this approach. For this reason, hydrogels were widely employed as tissue scaffolds because they can support cultivating cells and tissue in multi-dimensions. Major disadvantages of hydrogel-based cell cultivation include low mechanical properties, lack of topography, which should be enhanced for successful tissue engineering. Herein we surface-immobilized gelatin on the surface of nanofibrous matrix for 3-D cell cultivation in topographical cues added environments. Electrospun nanofibers were electrospun with injection of poly(caprolactone) through a single nozzle syringe. Electrospun meshes were then chopped up with a high speed grinder to fine powders. This was hydrolyzed in optimized concentration of sodium hydroxide solution from 1 to 6 hours and harvested by centrifugation. The freeze-dried powders were examined by scanning electron microscopy (SEM) for revealing the morphology and fibrilar shaped with a length of ca. 20um was observed. This was subsequently immersed in gelatin solution for surface-coating of gelatin, where the process repeated up to 10 times for obtaining desirable coating of gelatin on the surface. Gelatin-coated nanofibrils showed high waterswellability in comparison to the unmodified nanofibrils, and this enabled good dispersion properties of the modified nanofibrils in aqueous phase. The degree of water-swellability was increased as the coating numbers of gelatin increased, however, it did not any meaning result after 10 times of gelatin coating process. Thus, by adjusting the gelatin coating times, we could successfully control the degree of hydrophilicity and water-swellability of nanofibrils.

Keywords: nano, fiber, cell, tissue

Procedia PDF Downloads 143

Authors: Yu Chen, Seong-Jin Kim, Jaehoon Chung

Abstract:

High throughput cells counting and sizing are often required for biomedical applications. Here we report design, fabrication and validating of a micro-machined Coulter counter device with multiple-channel to realize such application for low cost. Multiple vertical through-holes were fabricated on a silicon chip, combined with the PDMS micro-fluidics channel that serves as the sensing channel. In order to avoid the crosstalk introduced by the electrical connection, instead of measuring the current passing through, the potential of each channel is monitored, thus the high throughput is possible. A peak of the output potential can be captured when the cell/particle is passing through the microhole. The device was validated by counting and sizing the polystyrene beads with diameter of 6 μm, 10 μm and 15 μm. With the sampling frequency to be set at 100 kHz, up to 5000 counts/sec for each channel can be realized. The counting and enumeration of MCF7 cancer cells are also demonstrated.

Keywords: Coulter counter, cell enumeration, high through-put, cell sizing

Procedia PDF Downloads 572

Authors: Samineh Barmaki, Ville Jokinen, Esko Kankuri

Abstract:

We report a microfluidic chip that can be used to modify the gaseous microenvironment of a cell-culture in ambient atmospheric conditions. The aim of the study is to show the cellular response to nitric oxide (NO) under hypoxic (oxygen < 5%) condition. Simultaneously targeting to hypoxic and nitric oxide will provide an opportunity for NO‑based therapeutics. Studies on cellular responses to lowered oxygen concentration or to gaseous mediators are usually carried out under a specific macro environment, such as hypoxia chambers, or with specific NO donor molecules that may have additional toxic effects. In our study, the chip consists of a microfluidic layer and a cell culture well, separated by a thin gas permeable polydimethylsiloxane (PDMS) membrane. The main design goal is to separate the gas oxygen scavenger and NO donor solutions, which are often toxic, from the cell media. Two different types of gas exchangers, titled 'pool' and 'meander' were tested. We find that the pool design allows us to reach a higher level of oxygen depletion than meander (24.32 ± 19.82 %vs -3.21 ± 8.81). Our microchip design can make the cells culture more simple and makes it easy to adapt existing cell culture protocols. Our first application is utilizing the chip to create hypoxic conditions on targeted areas of cell culture. In this study, oxygen scavenger sodium sulfite generates hypoxia and its effect on human embryonic kidney cells (HEK-293). The PDMS membrane was coated with fibronectin before initiating cell cultures, and the cells were grown for 48h on the chips before initiating the gas control experiments. The hypoxia experiments were performed by pumping of O₂-depleted H₂O into the microfluidic channel with a flow-rate of 0.5 ml/h. Image-iT® reagent as an oxygen level responser was mixed with HEK-293 cells. The fluorescent signal appears on cells stained with Image-iT® hypoxia reagent (after 6h of pumping oxygen-depleted H₂O through the microfluidic channel in pool area). The exposure to different levels of O₂ can be controlled by varying the thickness of the PDMS membrane. Recently, we improved the design of the microfluidic chip, which can control the microenvironment of two different gases at the same time. The hypoxic response was also improved from the new design of microchip. The cells were grown on the thin PDMS membrane for 30 hours, and with a flowrate of 0.1 ml/h; the oxygen scavenger was pumped into the microfluidic channel. We also show that by pumping sodium nitroprusside (SNP) as a nitric oxide donor activated under light and can generate nitric oxide on top of PDMS membrane. We are aiming to show cellular microenvironment response of HEK-293 cells to both nitric oxide (by pumping SNP) and hypoxia (by pumping oxygen scavenger solution) in separated channels in one microfluidic chip.

Keywords: hypoxia, nitric oxide, microenvironment, microfluidic chip, sodium nitroprusside, SNP

Procedia PDF Downloads 109

Authors: Jiraporn Rojtinnakorn

Abstract:

ICAM-2 (intercellular adhesion molecule 2) or CD102 (Cluster of Differentiation 102) is type I trans-membrane glycoproteins, composing 2-9 immunoglobulin-like C2-type domains. ICAM-2 plays the particular role in immune response and cell surveillance. It is concerned in innate and specific immunity, cell survival signal, apoptosis, and anticancer. EST clone of ICAM-2, from P. gigas blood cell EST libraries, showed high identity to human ICAM-2 (92%) with conserve region of ICAM N-terminal domain and part of Ig superfamily. Gene and protein of ICAM-2 has been founded in mammals. This is the first report of ICAM-2 in fish.

Keywords: ICAM-2, CD102, Pangasianodon gigas, antitumor

Procedia PDF Downloads 198

Authors: Kevser Dincer, Basma Waisi, M. Ozan Ozdemir, Ugur Pasaogullari, Jeffrey McCutcheon

Abstract:

Nanofibers are defined as fibers with diameters less than 100 nanometers. They can be produced by interfacial polymerization, electrospinning and electrostatic spinning. In this study, behaviours of activated carbon nano fiber (ACNF), carbon nano-fiber (CNF), Polyacrylonitrile/carbon nanotube (PAN/CNT), Polyvinyl alcohol/nano silver (PVA/Ag) in PEM fuel cells are investigated experimentally. This material was used as gas diffusion layer (GDL) in PEM fuel cells. When the performances of these cells are compared to each other at 5x5 cm2 cell, it is found that the PVA/Ag exhibits the best performance among all. In this work, nano fiber and nano fiber/nano particles electrical conductivities have been studied to understand their effects on PEM fuel cell performance. According to the experimental results, the maximum electrical conductivity performance of the fuel cell with nanofiber was found to be at PVA/Ag. The electrical conductivities of CNF, ACNF, PAN/CNT are lower for PEM. The resistance of cell with PVA/Ag is lower than the resistance of cell with PAN/CNT, ACNF, CNF.

Keywords: proton exchange membrane fuel cells, electrospinning, carbon nano fiber, activate carbon nano-fiber, PVA fiber, PAN fiber, carbon nanotube, nano particle nanocomposites

Procedia PDF Downloads 355

Authors: Dani Sukkar, Ali Kanso, Philippe Laval-Gilly, Jairo Falla-Angel

Abstract:

The honeybees' immune system is challenged by different risk factors that induce various responses. However, complex scenarios where bees are exposed to different pesticides simultaneously with immune activation are not well evaluated. The Toll pathway is one of the main signaling pathways studied in invertebrate immune responses, and it is a good indicator of the effect of such complex interactions in addition to key signaling elements of other pathways like Relish of the immune deficiency (IMD) pathway or Eater, the phagocytosis receptor or vitellogenin levels. Honeybee hemocytes extracted from 5th instar larvae were exposed to imidacloprid and/or amitraz with or without the presence of the zymosan a as an immune activator. The gene expression of multiple immune related genes were studied, including spaetzle, Toll, myD88, relish, eater and vitellogenin, by real-time polymerase chain reaction after RNA extraction. The results demonstrated that the Toll pathway is mainly affected by the pesticides; imidacloprid and amitraz, especially by their different combinations. Furthermore, immune activation by zymosan A, a fungal cell-wall component, acts to mitigate to some extent the effect of pesticides on the different levels of the Toll pathway. In addition, imidacloprid, amitraz, and zymosan A have complex and context-specific interactions depending on the levels of immune activation and the pathway evaluated affecting immune-gene expression differently.

Keywords: toll pathway, immune modulation, β-glucan, imidacloprid, amitraz, honeybees, immune genes

Procedia PDF Downloads 46

Authors: Kamta P. Namdeo

Abstract:

Novel thiazolidino-(3,2-b)-1, 2,4-triazole-5(6H)-one derivatives were synthesized, and anticancer activity was studied on human breast cancer cell line MFC7. It showed a significant decrease in cell viability with reference to the standard. The findings suggest that nitro-substituted compound showed best anticancer activity and activity was due to the triazole and thiazolidinone hetero nucleus present in the structure.

Keywords: anti-cancer, adriamycine, thiazolidinone, 1, 2, 4-triazole, thiazolidino-triazolone

Procedia PDF Downloads 343