Search results for: cells integration

Authors: Susanne Manes, Anni Glaeser, Katharina Wick, Bernhard Strauss, Uwe Berger

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Introduction: Over the last decades growing rates of mental health concerns among children and adolescents have been observed. At the same time, physical well-being of children and adolescents becomes increasingly impaired as well. Schools play an important role in preventing mental and physical disorders and in promoting well-being. Self-esteem, as well as social integration, are vital influence factors for mental and physical well-being. The purpose of this study was to develop and evaluate the program 'VorteilJena' for secondary schools in Germany focusing on self-esteem and social integration to improve mental and physical well-being. Method: The school-based health promotion program was designed for students in 5th grade and higher. It consists of several short pedagogical exercises instructed by a teacher and were integrated into the regular class over the course of ten weeks. The exercises focused on fostering social integration using either tasks improving team spirit or exercises that increase tolerance and sense of belonging. Other exercises focused on strengthening the self-esteem of the students. Additionally, the program included a poster exhibition titled 'Belonging' which was put up in the school buildings. The exhibition comprised ten posters which addressed relevant risk factors and resources related to social integration and self-esteem. The study was a randomized controlled sequential study with a pre and post measurement conducted in ten German schools. A total of 1642 students (44% male) were recruited. Their age ranged from 9 to 21 years (M=12.93 years; SD= 2.11). The program was conducted in classes ranging from 5th to 12th grade. Results: The program improved wellbeing, self-esteem and social integration of the involved students compared to the control group. Differential effects depending on implementation rates or age of the students will be analyzed. Moreover, implications for future school-based health promotion programs targeting self-esteem and social integration will be discussed. Conclusion: Social integration considerably influences self-esteem and well-being of students and can be targeted by school-based programs including short and modest exercises. Since a sufficient implementation of health promotion programs is essential, the present program due to its practicability represents a good opportunity to install health promotion focusing on social integration in schools.

Keywords: social integration, well-being, health promotion in schools, self-esteem

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Authors: Diane Vassallo., Leonard Busuttil

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Recent educational developments have seen increasing attention attributed to Computational Thinking (CT) and its integration into primary and secondary school curricula. CT is more than simply being able to use technology but encompasses fundamental Computer Science concepts which are deemed to be very important in developing the correct mindset for our future digital citizens. The case study presented in this article explores the journey of a Maltese secondary school teacher in his efforts to plan, develop and integrate CT within the context of a local classroom. The teacher participant was recruited from the Malta EU Code week summer school, a pilot initiative that stemmed from the EU Code week Team’s Train the Trainer program. The qualitative methodology involved interviews with the participant teacher as well as an analysis of the artefacts created by the students during the lessons. The results shed light on the numerous challenges and obstacles that the teacher encountered in his integration of CT, as well as portray some brilliant examples of good practices which can substantially inform further research and practice around the integration of CT in classroom practice.

Keywords: computational thinking, digital citizens, digital literacy, technology integration

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Authors: Tahereh Yavari, Sara Norozi Far

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The aim of this study was to compare psychological well-being, hope, and health concerns in leukemia patients before and after receiving stem cells. The statistical population of the present study was made up of leukemia patients in Tehran, and the research sample was among the patients referred to the Bone Marrow Transplant Center of Shariati Hospital in Tehran, and they were placed in two experimental and control groups (15 people in each group), which were selected by purposive sampling method. In order to collect the data for the research, three psychological well-being questionnaires were used by Riff (2002), Schneider's Hope Scale (SHS), and Schneider's Health Concern Questionnaire (HCQ). In order to analyze the data in this research, according to the "pre-test-post-test design with a control group," covariance analysis was used. Based on the research findings, it was concluded that receiving stem cells increases hope and psychological well-being in leukemia patients and significantly reduces health concerns.

Keywords: psychological well-being, hope, health concerns, blood cancer, stem cells

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Authors: S. Subramaniam, J. S. A. Rao, P. Ramdas, K. R. Selvaduray, N. M. Han, M. K. Kutty, A. K. Radhakrishnan

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Immune system is a complex system where the immune cells have the capability to respond against a wide range of immune challenges including cancer progression. However, in the event of cancer development, tumour cells trigger immunosuppressive environment via activation of myeloid-derived suppressor cells and T regulatory (Treg) cells. The Treg cells are subset of CD4+ T lymphocytes, known to have crucial roles in regulating immune homeostasis and promoting the establishment and maintenance of peripheral tolerance. Dysregulation of these mechanisms could lead to cancer progression and immune suppression. Recently, there are many studies reporting on the effects of natural bioactive compounds on immune responses against cancer. It was known that tocotrienol-rich-fraction consisting 70% tocotrienols and 30% α-tocopherol is able to exhibit immunomodulatory as well as anti-cancer properties. Hence, this study was designed to evaluate the effects of gamma-tocotrienol (G-T3) supplementation on T-reg cells in a syngeneic mouse model of breast cancer. In this study, female BALB/c mice were divided into two groups and fed with either soy oil (vehicle) or gamma-tocotrienol (G-T3) for two weeks followed by inoculation with tumour cells. All the mice continued to receive the same supplementation until day 49. The results showed a significant reduction in tumour volume and weight in G-T3 fed mice compared to vehicle-fed mice. Lung and liver histology showed reduced evidence of metastasis in tumour-bearing G-T3 fed mice. Besides that, flow cytometry analysis revealed T-helper cell population was increased, and T-regulatory cell population was suppressed following G-T3 supplementation. Moreover, immunohistochemistry analysis showed that there was a marked decrease in the expression of FOXP3 in the G-T3 fed tumour bearing mice. In conclusion, the G-T3 supplementation showed good prognosis towards breast cancer by enhancing the immune response in tumour-bearing mice. Therefore, gamma-T3 can be used as immunotherapy agent for the treatment of breast cancer.

Keywords: breast cancer, gamma tocotrienol, immune suppression, supplement

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Authors: D. J. Kalita

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Cancer is one of the prime causes of early death worldwide. Mutation of the gene involve in DNA repair and damage, like BRCA2 (Breast cancer gene two) genes, can be detected efficiently by PCR-RFLP to early breast cancer diagnosis and adopt the suitable method of treatment. Host Defense Peptide can be used as blueprint for the design and synthesis of novel anticancer drugs to avoid the side effect of conventional chemotherapy and chemo resistance. The change at nucleotide position 392 of a -› c in the cancer sample of dog mammary tumour at BRCA2 (exon 7) gene lead the creation of a new restriction site for SsiI restriction enzyme. This SNP may be a marker for detection of canine mammary tumour. Support vector machine (SVM) algorithm was used to design and predict the anticancer peptide from the mature functional peptide. MTT assay of MCF-7 cell line after 48 hours of post treatment showed an increase in the number of rounded cells when compared with untreated control cells. The ability of the synthesized peptide to induce apoptosis in MCF-7 cells was further investigated by staining the cells with the fluorescent dye Hoechst stain solution, which allows the evaluation of the nuclear morphology. Numerous cells with dense, pyknotic nuclei (the brighter fluorescence) were observed in treated but not in control MCF-7 cells when viewed using an inverted phase-contrast microscope. Thus, PCR-RFLP is one of the attractive approach for early diagnosis, and synthetic cationic peptide can be used for the treatment of canine mammary tumour.

Keywords: cancer, cationic peptide, host defense peptides, Breast cancer genes

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Authors: Hülya Kuru Mutlu, Mustafa Kulakcı, Uğur Serincan

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The sun is one of the latest developments in renewable energy sources, which has a variety of application. Solar energy is the most preferred renewable energy sources because it can be used directly, it protects the environment and it is economic. In this work, we investigated that important parameter of GaAs-based solar cells with respect to the growth temperature. The samples were grown on (100) oriented p-GaAs substrates by solid source Veeco GEN20MC MBE system equipped with Ga, In, Al, Si, Be effusion cells and an Arsenic cracker cell. The structures of the grown samples are presented. After initial oxide desorption, Sample 1 and Sample 2 were grown at about 585°C and 535°C, respectively. From the grown structures, devices were fabricated by using the standard photolithography procedure. Current-voltage measurements were performed at room temperature (RT). It is observed that Sample 1 which was grown at 585°C has higher efficiency and fill factor compared to Sample 2. Hence, it is concluded that the growth temperature of 585°C is more suitable to grow GaAs-based solar cells considering our samples used in this study.

Keywords: molecular beam epitaxy, solar cell, current-voltage measurement, Sun

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Authors: Pintusorn Hansakul, Ruchilak Rattarom, Arunporn Itharat

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Background: Benjakul, a Thai traditional herbal formulation, comprises of five plants: Piper chaba, Piper sarmentosum, Piper interruptum, Plumbago indica, and Zingiber officinale. It has been widely used to treat cancer patients in the context of folk medicine in Thailand. This study aimed to investigate the cytotoxic effect of the ethanol extract of Benjakul against three non-small cell lung cancer (NSCLC) cell lines (NCI-H226, A549, COR-L23), small cell lung cancer (SCLC) cell line NCI-H1688 and normal lung fibroblast cell line MRC-5. The study further examined the molecular mechanisms underlying its cytotoxicity via induction of apoptosis in NCI-H226 cells. Methods: The cytotoxic effect of Benjakul was determined by SRB assay. The effect of Benjakul on cell cycle distribution was assessed by flow cytometric analysis. The apoptotic effects of Benjakul were determined by sub-G1 quantitation and Annexin V-FITC/PI flow cytometric analyses as well as by changes in caspase-3 activity. Results: Benjakul exerted potent cytotoxicity on NCI-H226 and A549 cells but lower cytotoxicity on COR-L23 and NCI-H1688 cells without any cytotoxic effect on normal cells. Molecular studies showed that Benjakul extract induced G2/M phase arrest in human NCI-H226 cells in a dose-dependent manner. The highest concentration of Benjakul (150 μg/ml) led to the highest increase in the G2/M population at 12 h, followed by the highest increase in the sub-G1 population (apoptotic cells) at 60 h. Benjakul extract also induced early apoptosis (AnnexinV +/PI−) in NCI-H226 cells in a dose- and time- dependent manner. Moreover, treatment with 150 μg/ml Benjakul extract for 36 h markedly increased caspase-3 activity by 3.5-fold, and pretreatment with the general caspase inhibitor z-VAD-fmk completely abolished such activity. Conclusions: This study reveals for the first time the regulation of apoptosis in human lung cancer NCI-H226 cells through caspase-dependent mechanism by Benjakul extract.

Keywords: apoptosis, Benjakul, caspase activation, cytotoxicity

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Authors: Shuenn-Yih Chang, Chiu-LI Huang, Ngoc-Cuong Tran

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A new family of structure-dependent integration methods, whose coefficients of the difference equation for displacement increment are functions of the initial structural properties and the step size for time integration, is proposed in this work. This family method can simultaneously integrate the controllable numerical dissipation, explicit formulation and unconditional stability together. In general, its numerical dissipation can be continuously controlled by a parameter and it is possible to achieve zero damping. In addition, it can have high-frequency damping to suppress or even remove the spurious oscillations high frequency modes. Whereas, the low frequency modes can be very accurately integrated due to the almost zero damping for these low frequency modes. It is shown herein that the proposed family method can have exactly the same numerical properties as those of HHT-α method for linear elastic systems. In addition, it still preserves the most important property of a structure-dependent integration method, which is an explicit formulation for each time step. Consequently, it can save a huge computational efforts in solving inertial problems when compared to the HHT-α method. In fact, it is revealed by numerical experiments that the CPU time consumed by the proposed family method is only about 1.6% of that consumed by the HHT-α method for the 125-DOF system while it reduces to be 0.16% for the 1000-DOF system. Apparently, the saving of computational efforts is very significant.

Keywords: structure-dependent integration method, nonlinear dynamic analysis, unconditional stability, numerical dissipation, accuracy

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Authors: Ali A. Alshatwi, Vaiyapuri S. Periasamy, Jegan Athinarayanan

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Engineered nanoparticles’ usage rapidly increased in various applications in the last decade due to their unusual properties. However, there is an ever increasing concern to understand their toxicological effect in human health. Particularly, metal and metal oxide nanoparticles have been used in various sectors including biomedical, food and agriculture. But their impact on human health is yet to be fully understood. In this present investigation, we assessed the toxic effect of engineered nanoparticles (ENPs) including Ag, MgO and Co3O4 nanoparticles (NPs) on human mesenchymal stem cells (hMSC) adopting cell viability and cellular morphological changes as tools The results suggested that silver NPs are more toxic than MgO and Co3O4NPs. The ENPs induced cytotoxicity and nuclear morphological changes in hMSC depending on dose. The cell viability decreases with increase in concentration of ENPs. The cellular morphology studies revealed that ENPs damaged the cells. These preliminary findings have implications for the use of these nanoparticles in food industry with systematic regulations.

Keywords: cobalt oxide, human mesenchymal stem cells, MgO, silver

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Authors: Pelin Balcik Ercin

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Introduction: ZEB transcription factor family member ZEB2, has a role in epithelial to mesenchymal transition during development and metastasis. The altered circulating extracellular miRNAs expression is observed in diseases, and extracellular miRNAs have an important role in cancer cell microenvironment. In ChIP-Seq study, the expression of miR-2110 was found to be regulated by ZEB2. In this study, the effects of miR2110 on cell proliferation and migration of hepatocellular carcinoma (HCC) cells were examined. Material and Methods: SNU398 cells transfected with mimic miR2110 (20nM) (HMI0375, Sigma-Aldrich) and negative control miR (HMC0002, Sigma-Aldrich). MicroRNA isolation was accomplished with miRVANA isolation kit according to manufacturer instructions. cDNA synthesis was performed expression, respectively, and calibrated with Ct of controls. The real-time quantitative PCR (RT-qPCR) reaction was performed using the TaqMan Fast Advanced Master Mix (Thermo Sci.). Ct values of miR2110 were normalized to miR-186-5p and miR16-5p for the intracellular gene. Cell proliferation analysis was analyzed with the xCELLigence RTCA System. Wound healing assay was analyzed with the ImageJ program and relative fold change calculated. Results: The mimic-miR-2110 transfected SNU398 cells nearly nine-fold (log2) more miR-2110 expressed compared to negative control transfected cells. The mimic-miR-2110 transfected HCC cell proliferation significantly inhibited compared to the negative control cells. Furthermore, miR-2110-SNU398 cell migration capacity was relatively four-fold decreased compared to negative control-miR-SNU398 cells. Conclusion: Our results suggest the miR-2110 inhibited cell proliferation and also miR-2110 negatively affect cell migration compared to control groups in HCC cells. These data suggest the complexity of microRNA EMT transcription factors regulation. These initial results are pointed out the predictive biomarker capacity of miR-2110 in HCC.

Keywords: epithelial to mesenchymal transition, EMT, hepatocellular carcinoma cells, micro-RNA-2110, ZEB2

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Authors: Amina Ben Abla, Sylvie Changotade, Geraldine Rohman, Guilhem Boeuf, Cyrine Dridi, Ahmed Elmarjou, Florence Dufour, Didier Lutomski, Abdellatif Elm’semi

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Understanding cell adhesion and interaction with the extracellular matrix is essential for biomedical and biotechnological applications, including the development of biomaterials. In recent years, numerous biomaterials have emerged and were used in the field of tissue engineering. Nevertheless, the lack of interaction of biomaterials with cells still limits their bio-integration. Thus, the design of bioactive biomaterials to improve cell attachment and proliferation is of growing interest. In this study, bio-functionalized material was developed combining a synthetic polymer scaffold surface with selected domains of type III human fibronectin (FNIII-DOM) to promote cell adhesion and proliferation. Bioadhesive ligand includes cell-binding domains of human fibronectin, a major ECM protein that interacts with a variety of integrins cell-surface receptors, and ECM proteins through specific binding domains were engineered. FNIII-DOM was produced in bacterial system E. coli in 5L fermentor with a high yield level reaching 20mg/L. Bioactivity of the produced fragment was validated by studying cellular adhesion of human cells. The adsorption and immobilization of FNIII-DOM onto the polymer scaffold were evaluated in order to develop an innovative biomaterial.

Keywords: biomaterials, cellular adhesion, fibronectin, tissue engineering

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Authors: Jayesh M. Sonawane, Prakash C. Ghosh

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Landfill leachate emerges as a promising feedstock for microbial fuel cells (MFCs). In the present investigation, direct air-breathing cathode-based MFCs are fabricated to investigate the potential of landfill leachate. Three MFCs that have different cathode areas are fabricated and investigated for 17 days under open circuit conditions. The maximum open circuit voltage (OCV) is observed to be as high as 1.29 V. The maximum cathode area specific power density achieved in the reactor is 1513 mW m^-2. Further studies are under progress to understand the origin of high OCV obtained from landfill leachate-based MFCs.

Keywords: microbial fuel cells, landfill leachate, air-breathing cathode, performance study

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Authors: Leila Ayat, Afak Meftah

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Amorphous-silicon alloys have great promise as low cost solar cell materials. They have excellent photo-conductivity and high optical absorption to sunlight. Now PIN a-Si:H based solar cells are widely used in power generation modules. However, to improve the performance of these cells further, a better fundamental under-standing of the factors limiting cell performance in the homo junction PIN structure is necessary. In this paper we discuss the sensitivity of light J-V characteristics to various device and material parameters in PIN homo junction solar cells. This work is a numerical simulation of the output parameters of a PIN a-Si:H solar cell under AM1.5 spectrum. These parameters are the short circuit current (Jsc), the open circuit voltage (Voc), the fill factor (FF), the conversion efficiency. The simulation was performed with SCAPS-1D software version 3.3 developed at ELIS in Belgium by Marc Burgelman et al. The obtained results are in agreement with experiment. In addition, the effect of the thickness, doping density, capture cross sections of the gap states and the band microscopic mobilities on the output parameters of the cell are also presented.

Keywords: amorphous silicon p-i-n junctions, thin film, solar cells, sensitivity

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Authors: Hien Thi Thu Le, Nuno Rafael Candeias, Olli Yli-Harja, Meenakshisundaram Kandhavelu

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Purinergic receptor 1 (P2Y1R) is the potential therapeutic target for inducing prostate cancer (PCa) cell death. Recently, 1-indolinoalkyl 2-phenolic derivative, HIC, was identified as a P2Y1R agonist that increases apoptosis and inhibits cell proliferation of PCa. However, the biological effects of HIC have not been extensively studied at the molecular level. In the present study, we have investigated the anticancer effects of HIC and the molecular mechanisms underlying in PCa cells. Half maximal inhibitory concentration (IC₅₀) of HIC was measured as 15.98 μM and 15.64 μM for DU145 and PC3 cells, respectively. In addition, we found that HIC inhibited cell growth and metastasis of PC3 and DU145 cells colonies, spheroid areas, and migrated cells. RNA seq analysis revealed significant changes of over 3000 genes (p value < 0.05) upon HIC treatment in PC3 and DU145 cells. Genes involved in DNA damage, apoptosis, cell cycle arrest at G1/S phase were modulated by HIC treatment. MAPK and NF-κB protein array revealed the increased expression of ERK1/2, JNK1/2, p53 phosphorylation, and p53 protein. ERK1/2 and JNK1/2 activations are known to increase the stabilization of p53, a tumor suppressor protein, which is required to arrest the cell cycle at G1/S phase and cause cell death of PCa cells. Overall, our results suggest that HIC can serve as a multi-dimensional chemotherapeutic agent possessing strong cytotoxic, anti-cancer, and anti-metastasis against PCa growth.

Keywords: prostate cancer, P2Y1 receptor, apoptosis, metastasis

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Authors: Bekir Turedi

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Exploiting the potential of perovskite single-crystal solar cells in optoelectronic applications necessitates overcoming a significant challenge: the low charge collection efficiency at increased thickness, which has restricted their deployment in radiation detectors and nuclear batteries. Our research details a promising approach to this problem, wherein we have successfully fabricated single-crystal MAPbI3 solar cells employing a space-limited inverse temperature crystallization (ITC) methodology. Remarkably, these cells, up to 400-fold thicker than current-generation perovskite polycrystalline films, maintain a high charge collection efficiency even without external bias. The crux of this achievement lies in the long electron diffusion length within these cells, estimated to be around 0.45 mm. This extended diffusion length ensures the conservation of high charge collection and power conversion efficiencies, even as the thickness of the cells increases. Fabricated cells at 110, 214, and 290 µm thickness manifested power conversion efficiencies (PCEs) of 20.0, 18.4, and 14.7% respectively. The single crystals demonstrated nearly optimal charge collection, even when their thickness exceeded 200 µm. Devices of thickness 108, 214, and 290 µm maintained 98.6, 94.3, and 80.4% of charge collection efficiency relative to their maximum theoretical short-circuit current value, respectively. Additionally, we have proposed an innovative, self-consistent technique for ascertaining the electron-diffusion length in perovskite single crystals under operational conditions. The computed electron-diffusion length approximated 446 µm, significantly surpassing previously reported values for this material. In conclusion, our findings underscore the feasibility of fabricating halide perovskite single-crystal solar cells of hundreds of micrometers in thickness while preserving high charge extraction efficiency and PCE. This advancement paves the way for developing perovskite-based optoelectronics necessitating thicker active layers, such as X-ray detectors and nuclear batteries.

Keywords: perovskite, solar cell, single crystal, diffusion length

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Authors: M. S. Hasni, J. Guan, K. Yakimchuk, M. Berglund, B. Sander, G. Enblad, R. M. Amini, S. Okret

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Lymphomas are generally not considered as endocrine-related cancers. However, most lymphoid malignancies show gender differences in incidence and show prognosis with males being more affected. Furthermore, some epidemiological data indicate a protective role of estrogens against Non-Hodgkin lymphomas. Recent studies have demonstrated estrogen receptor β (ERβ) to be the major ER expressed in normal and malignant cells of lymphoid origin. We have analyzed the effects of estradiol and selective ERα and ERβ agonists on lymphoma growth in culture and in vivo. Treating lymphoma cells with estradiol or ERα selective agonist had minor or no effect on cell growth while selective ERβ agonist treatment showed an antiproliferative effect. When grafting mice with murine T lymphoma cells, male mice developed larger tumors compared to female mice, a difference that was abolished following ovariectomy, demonstrating estrogen-dependent growth in vivo. When subcutaneously grafting lymphoma cells to mice, so far growth of all tested human B lymphoma tumors (Raji and Ramos Burkitt lymphoma, SU.DHL4 (GC) and U2932 (ABC) DLBCL, Granta-519, Maver1 and Z138 MCL cells), were reduced following treatment with ERβ selective agonist (ref. 2 and unpublished). Moreover, the number and size of liver foci of disseminating Raji cells was reduced. We have identified target genes and mechanism that could explain the above effects of ERβ agonists. This included effects on angio and lymphangiogenesis. Now we have further analyzed effects of ERβ agonists on Ibrutinib-sensitive and -insensitive MCL cells in xenograft experiments as well as ERβ expression in primary lymphoma material (DLBCL). Preliminary statistical analysis has been done correlating ERβ expression to other biomarkers and clinical data.

Keywords: lymphomas, estrogen receptors, cancer, liver foci

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Authors: Kkochorong Park, Keun Cheon Kim, Hyoban Lee, Eun Ju Lee, Bongsoo Kim

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Introduction of biomaterials such as DNA, RNA, proteins is important for many research areas. There are many methods to introduce biomaterials into living organisms like tissue and cells. To introduce biomaterials, several indirect methods including virus‐mediated delivery, chemical reagent (i.e., lipofectamine), electrophoresis have been used. Such methods are passive delivery using an endocytosis process of cell, reducing an efficiency of delivery. Unlike the indirect delivery method, it has been reported that a direct delivery of exogenous biomolecules into nucleus have been more efficient to expression or integration of biomolecules. Nano-sized material is beneficial for detect signal from cell or deliver stimuli/materials into the cell at cellular and molecular levels, due to its similar physical scale. Especially, because 1 dimensional (1D) nanomaterials such as nanotube, nanorod and nanowire with high‐aspect ratio have nanoscale geometry and excellent mechanical, electrical, and chemical properties, they could play an important role in molecular and cellular biology. In this study, by using single crystalline 1D noble metal nanowire, we fabricated nano-sized 1D injector which can successfully interface with living cells and directly deliver biomolecules into several types of cell line (i.e., stem cell, mammalian embryo) without inducing detrimental damages on living cell. This nano-bio technology could be a promising and robust tool for introducing exogenous biomaterials into living organism.

Keywords: DNA, gene delivery, nanoinjector, nanowire

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Authors: Natalia Koltovaya, Nadezhda Zhuchkina, Ksenia Lyubimova

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We study the biological effects induced by ionizing radiation in view of therapeutic exposure and the idea of space flights beyond Earth's magnetosphere. In particular, we examine the differences between base pair substitution induction by ionizing radiation in model haploid and diploid yeast Saccharomyces cerevisiae cells. Such mutations are difficult to study in higher eukaryotic systems. In our research, we have used a collection of six isogenic trp5-strains and 14 isogenic haploid and diploid cyc1-strains that are specific markers of all possible base-pair substitutions. These strains differ from each other only in single base substitutions within codon-50 of the trp5 gene or codon-22 of the cyc1 gene. Different mutation spectra for two different haploid genetic trp5- and cyc1-assays and different mutation spectra for the same genetic cyc1-system in cells with different ploidy — haploid and diploid — have been obtained. It was linear function for dose-dependence in haploid and exponential in diploid cells. We suggest that the differences between haploid yeast strains reflect the dependence on the sequence context, while the differences between haploid and diploid strains reflect the different molecular mechanisms of mutations.

Keywords: base pair substitutions, γ-rays, haploid and diploid cells, yeast Saccharomyces cerevisiae

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Authors: Priyanka Chowdhury, Asitikantha Sarma, Utpal Ghosh

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Hadron therapy using high Linear Energy Transfer (LET) ion beam is producing promising clinical results worldwide. The major advantages are its ability to kill radio-resistant tumor and its anti-metastatic activity. Poly(ADP-ribose) polymerase-1 (PARP-1) inhibitors have been widely used as radiosensitizer, but its role in metastasis is unknown. The purpose of our study was to investigate the effect of PARP-1 depletion in combination with either Carbon Ion Beam (CIB) or gamma irradiation on metastatic potential of cultured cancerous cells. A549 cells were irradiated with CIB (0-4Gy) or gamma (0, 2, 4, 6 and 10 Gy) with and without PARP-1 inhibition. The metastatic potential of the cells was determined by cell migratory assay, expression, and activity of MMP-2 and MMP-9, expression of Cadherin, Fibronectin, and Vimentin. CIB exposure reduced migratory property and activity of MMP-2 and MMP-9 significantly. CIB with PARP-1 inhibition reduced cell migration and Matrix Metalloproteinase (MMPs) activity in a synergistic manner. Expression of MMPs was also down-regulated in CIB and combined treatment. On the contrary, MMP- 2 and MMP-9 activity was significantly increased in gamma irradiated cells but decreased upon combined treatment of gamma and PARP-1 inhibitor. MMPs expression and migration was reduced when gamma irradiation was combined with PARP-1 inhibition. Thus, our study clearly demonstrates that PARP-1 inhibition in combination with either high or low LET can significantly suppress metastatic potential in cancer cells and thereby can be a promising tool in controlling metastatic cancers.

Keywords: high LET, low LET, matrix metalloproteinase (MMP), PARP-1

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Authors: Naif Al-Jomah, Falah H Al-Mohanna, Abdelilah Aboussekhra

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Active breast cancer-associated fibroblasts (CAFs), the most influential cells in breast tumor microenvironment, express/secrete high levels of the proinvasive/metastatic interleukin-6 (IL-6). Therefore, we have tested here the effect of the IL-6 receptor (IL-6R) inhibitor tocilizumab (TCZ; Actemra) on different active breast CAFs. We have shown that TCZ potently and persistently suppresses the expression of various CAF biomarkers, namely α-SMA, SDF-1 as well as the STAT3 pathway and its downstream target AUF1. TCZ also inhibited the proliferation, migration and invasion abilities of active breast CAF cells. Additionally, TCZ repressed the ability of CAF cells in promoting epithelial-to-mesenchymal transition, and enhancing the migratory/invasive and proliferative capacities of breast cancer cells in vitro. Importantly, these findings were confirmed in orthotopic humanized breast tumors in mice. Furthermore, TCZ suppressed the expression of the pro-angiogenic factor VEGF-A and its transactivator HIF-1α in CAF cells, and consequently inhibited the angiogenic-promoting effect of active CAFs both in vitro and in orthotopic tumor xenografts. These results indicate that inhibition of the IL-6/STAT3/AUF1 pathway by TCZ can normalize active breast CAFs and suppress their paracrine pro-carcinogenic effects, which paves the way toward development of specific CAF-targeting therapy, badly needed for more efficient breast cancer treatments.

Keywords: angiogenesis, interleukin-6, paracrine, cancer-associated fibroblasts

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Authors: Ahmed Moulay, Mariia Zhuldybina, Mirko Torres, Mike Rozel, Ngoc Duc Trinh, Chloé Bois

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Hybrid printed electronics technology (HPE) provides innovative opportunities to enhance conventional electronics applications, which are often based on printed circuit boards (PCB). By combining the best of both performance from conventional electronic components and the flexibility from printed circuits makes it possible to manufacture HPE at high volumes using roll-to-roll printing processes. However, several challenges must be overcome in order to accurately integrate an electronic component on a printed circuit. In this presentation, we will demonstrate the integration process of electronic components from the lab scale to the industrialization. Both the printing quality and the integration technique must be studied to define the optimal conditions. To cover the parameters that influence the print quality of the printed circuit, different printing processes, flexible substrates, and conductive inks will be used to determine the optimized printing process/ink/substrate system. After the systems is selected, an electronic component of 2.5 mm2 chip size will be integrated to validate the functionality of the printed, electronic circuit. Critical information such as the conductive adhesive, the curing conditions, and the chip encapsulation will be determined. Thanks to these preliminary results, we are able to demonstrate the chip integration on a printed circuit using industrial equipment, showing the potential of industrialization, compatible using roll-to-roll printing and integrating processes.

Keywords: flat bed screen-printing, hybrid printed electronics, integration, large-scale production, roll-to-roll printing, rotary screen printing

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Authors: Hager E. Amer, Hany El Saftawy, Laila Rashed, Ahmed M. Ata, Fatma Metwally, Hesham Mettawei, Hossam E. Sayed, Tamer Adel, Kareem M. El Sawah

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Aim: The purpose of this study is to regenerate the damaged photoreceptor cells as a result of argon laser as a model of Retinitis Pigmentosa in rabbits' retina by using adult stem cells from rabbits' bone marrow. Background: Retinitis pigmentosa (RP) is a group of inherited disorders that primarily affect photoreceptor and pigment epithelium function. RP leads to loss of the rod outer segment and shorten the photoreceptor layer and expose the photoreceptor cell body to high-pressure levels in oxygen (oxidative stress) leads to apoptosis to the rod and cone cells. In particular, there is no specific treatment for retinitis pigmentosa. Materials and Methods: Forty Two Giant (Rex) rabbits were used in this experiment divided into 3 groups: Group 1: Control (6 rabbits), Group 2: Argon laser radiated as a model of retinitis pigmentosa (12 rabbits), Group 3: Laser radiated and treated by 6 million stem cells (12 rabbits). The last two groups are divided each into two subgroups each subgroup contains 6 rabbits, the ophthalmological examination was performed on rabbits, blood samples and retina samples were taken after 25 days and after 36 days from the laser radiation (10 days and 21 days after stem cells insertion in group 3) to perform the biochemical analysis. Results: Compared to control Group, a decrease of ERG wave amplitude and antioxidant substances (Glutathione) in blood and retina in group 2, and an increase of oxidative stress substances (Nitric oxide, Malonaldehyde, and carponyl protein) and apoptotic substances (Advanced glycation end product and M-metalloproteinase) in blood and retina. Compared to group 2, mostly increases of antioxidant substances and ERG wave amplitude in group 3, and mostly decreases in oxidative stress substances and apoptotic substances. Conclusion: Insertion of 6 million stem cells intravitreous gives good results in regeneration of the damaged photoreceptor cells after 21 days.

Keywords: retinitis pigmentosa, stem cells, argon laser, oxidative stress, apoptosis

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Authors: Haitham S. Hamza, Mohamed Maher, Shourok Alaa, Aya Khattab, Hadeal Ismail, Kamilia Hosny

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Radio Frequency Identification (RFID) has become a key technology in the margining concept of Internet of Things (IoT). Naturally, business applications would require the deployment of various RFID systems that are developed by different vendors and use various data formats. This heterogeneity poses a real challenge in developing large-scale IoT systems with RFID as integration is becoming very complex and challenging. Semantic integration is a key approach to deal with this challenge. To do so, ontology for RFID systems need to be developed in order to annotated semantically RFID systems, and hence, facilitate their integration. Accordingly, in this paper, we propose ontology for RFID systems. The proposed ontology can be used to semantically enrich RFID systems, and hence, improve their usage and reasoning. The usage of the proposed ontology is explained through a simple scenario in the health care domain.

Keywords: RFID, semantic technology, ontology, sparql query language, heterogeneity

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Authors: Emily Schlebes, Christian Hundhausen, Jens W. Fischer

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The glycosaminoglycan hyaluronan (HA) is a major component of the extracellular matrix, typically produced by fibroblasts of the connective tissue but also by immune cells. Here, we investigated the capacity of human peripheral blood CD8 T cells from healthy donors to produce HA and to express HA receptors as well as HA degrading enzymes. Further, we evaluated the effect of pharmacological HA inhibition on CD8 T cell function. Using immunocytochemistry together with quantitative PCR analysis, we found that HA synthesis is rapidly induced upon antibody-induced T cell receptor (TCR) activation and almost exclusively mediated by HA synthase 3 (HAS3). TCR activation also resulted in the upregulation of HA receptors CD44, hyaluronan-mediated motility receptor (HMMR), and layilin (LAYN), although kinetics and strength of expression varied greatly between subjects. The HA-degrading enzymes HYAL1 and HYAL2 were detected at low levels and induced by cell activation in some individuals. Interestingly, expression of HAS3, HA receptors, and hyaluronidases were modulated by the proinflammatory cytokines IL-6 and IL-1bβ in most subjects. To assess the functional role of HA in CD8 T cells, we performed carboxyfluorescein succinimidyl ester (CFSE) based proliferation assays and cytokine analysis in the presence of the HA inhibitor 4- Methylumbelliferone (4-MU). Despite significant inter-individual variation with regard to the effective dose, 4-MU resulted in the inhibition of CD8 T cell proliferation and reduced release of TNF-α and IFN-γ. Collectively, these data demonstrate that human CD8 T cells respond to TCR stimulation with a synthesis of HA and expression of HA-related genes. They further suggest that HA inhibition may be helpful in interfering with pathogenic T cell activation in human disease.

Keywords: CD8 T cells, extracellular matrix, hyaluronan, hyaluronan synthase 3

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Authors: Archana Sharma, Vijayashree Nayak, Ashok Kumar

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Three-dimensional matrices were fabricated from blend of natural-natural polymers like carrageenan-gelatin and synthetic -natural polymers such as PEG- gelatin (PEG of different molecular weights (2,000 and 6,000) using two different crosslinkers; glutaraldehyde and EDC-NHS by cryogelation technique. Blends represented a feasible approach to design 3-D scaffolds with controllable mechanical, physical and biochemical properties without compromising biocompatibility and biodegradability. These matrices possessed interconnected porous structure, good mechanical strength, biodegradable nature, constant swelling kinetics, ability to withstand high temperature and visco-elastic behavior. Hemocompatibility of cryogel matrices was determined by coagulation assays and hemolytic activity assay which demonstrated that these cryogels have negligible effects on coagulation time and have excellent blood compatibility. In vitro biocompatibility (cell-matrix interaction) inferred good cell adhesion, proliferation, and secretion of ECM on matrices. These matrices provide a microenvironment for the growth, proliferation, differentiation and secretion of ECM of different cell types such as IMR-32, C2C12, Cos-7, rat bone marrow derived MSCs and human bone marrow MSCs. Hoechst 33342 and PI staining also confirmed that the cells were uniformly distributed, adhered and proliferated properly on the cryogel matrix. An ideal scaffold used for tissue engineering application should allow the cells to adhere, proliferate and maintain their functionality. Neurotransmitter analysis has been done which indicated that IMR-32 cells adhered, proliferated and secreted neurotransmitters when they interacted with these matrices which showed restoration of their functionality. The cell-matrix interaction up to molecular level was also evaluated so to check genotoxicity and protein expression profile which indicated that these cryogel matrices are non-genotoxic and maintained biofunctionality of cells growing on these matrices. All these cryogels, when implanted subcutaneously in balb/c mice, showed no adverse systemic or local toxicity effects at implantation site. There was no significant increase in inflammatory cell count has otherwise been observed after scaffold implantation. These cryogels are supermacroporous and this porous structure allows cell infiltration and proliferation of host cells. This showed the integration and presence of infiltrated cells into the cryogel implants. Histological analysis confirmed that the implanted cryogels do not have any adverse effect in spite of host immune system recognition at the site of implantation, on its surrounding tissues and other vital host organs. In vivo biocompatibility study after in vitro biocompatibility analysis has also concluded that these synthesized cryogels act as important biological substitutes, more adaptable and appropriate for transplantation. Thus, these cryogels showed their potential for soft tissue engineering applications.

Keywords: cryogelation, hemocompatibility, in vitro biocompatibility, in vivo biocompatibility, soft tissue engineering applications

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Authors: Halil Ibrahim Demir, Alper Goksu, Onur Canpolat, Caner Erden, Melek Nur

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Traditionally process planning, scheduling and due date assignment are performed sequentially and separately. High interrelation between these functions makes integration very useful. Although there are numerous works on integrated process planning and scheduling and many works on scheduling with due date assignment, there are only a few works on the integration of these three functions. Here we tested the different integration levels of these three functions and found a fully integrated version as the best. We applied genetic search and random search and genetic search was found better compared to the random search. We penalized all earliness, tardiness and due date related costs. Since all these three terms are all undesired, it is better to penalize all of them.

Keywords: process planning, scheduling, due-date assignment, genetic algorithm, random search

Procedia PDF Downloads 357

Authors: Luciana Aparecida Bastos, Tatiana Diair L. F. Rosa, Jesus Creapldi

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The idea of Latin American integration occurred from the ideals of Simón Bolívar that, in 1824, called the Ibero-American nations to Amphictyonic Congress of Panama, on June 22, 1826, where he would defend the importance of Latin American unity. However, this congress was frustrating and the idea of Bolívar went no further. It was only after the European Union to start the process, driven by the end of World War II that the subject returned to emerge in Latin America. Thus, in 1960, supported by the European integration process, started in 1957 with the excellent result of the ECSC - European Coal and Steel Community, a result of the Customs Union of the BENELUX (integration between Belgium, the Netherlands and Luxembourg) in 1948, was created in Latin America, LAFTA - Latin American Free Trade Association, in 1960. In 1980, LAFTA was replaced by LAAI- Latin American Association, both with the same goal: to integrate Latin America, it´s economy and its trade. Most researchers in this period agree that the regional market would be expanded through the integration. The creation of one or more economic blocs in the region would provide the union of Latin American countries through a fusion of common interests and by their geographical proximity, which would try to develop common projects to promote mutual growth and economic development, tariff reductions, promotion of increased trade between, among many other goals set together. Thus, taking into account Mercosur, the main Latin-American block, created in 1994, the aim of this paper is to make a brief analysis of the trade balance performance of Brazil (larger economy of the block) in Mercosur in the periods: 1989-1994 and 1994-2012. The choice of this period was because the objective is to compare the period before and after the integration of Brazil in Mercosur. The methodologies used were the literature review and descriptive statistics. The results showed that after the integration of Brazil in Mercosur, the exports and imports grew within the bloc and the country turned out to become the leading importer of other economies of Mercosur after integration, that is, Brazil, after integration to Mercosur, was largely responsible for promoting the expansion of regional trade through the import of products from other members of the block.

Keywords: Brazil, mercosur, integration, trade balance, comparison

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Authors: Federico Herrera, Valle Gomez García de Soria, Itxaso Portero Sainz, Carlos Fernández Arandojo, Mercedes Royg, Ana Marcos Jimenez, Anna Kreutzman, Cecilia MuñozCalleja

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Introduction: Graft-versus-host disease (GVHD) still remains the major complication associated with allogeneic stem cell transplantation (SCT). The pathogenesis involves migration of donor naïve T-cells into recipient secondary lymphoid organs. Two molecules are important in this process: CD62L and CCR7, which are characteristically expressed in naïve/central memory T-cells. With this background, we aimed to study the influence of CCR7 and CD62L on donor lymphocytes in the development and severity of GVHD. Material and methods: This single center study included 98 donor-recipient pairs. Samples were collected prospectively from the apheresis product and phenotyped by flow cytometry. CCR7 and CD62L expression in CD4+ and CD8+ T-cells were compared between patients who developed acute (n=40) or chronic GVHD (n=33) and those who did not (n=38). Results: The patients who developed acute GVHD were transplanted with a higher percentage of CCR7+CD4+ T-cells (p = 0.05) compared to the no GVHD group. These results were confirmed when these patients were divided in degrees according to the severity of the disease; the more severe disease, the higher percentage of CCR7+CD4+ T-cells. Conversely, chronic GVHD patients received a higher percentage of CCR7+CD8+ T-cells (p=0.02) in comparison to those who did not develop the complication. These data were also confirmed when patients were subdivided in degrees of the disease severity. A multivariable analysis confirmed that percentage of CCR7+CD4+ T-cells is a predictive factor of acute GVHD whereas the percentage of CCR7+CD8+ T-cells is a predictive factor of chronic GVHD. In vitro functional assays (migration and activation assays) supported the idea of CCR7+ T-cells were involved in the development of GVHD. As low levels of CD62L expression were detected in all apheresis products, we tested the hypothesis that CD62L was shed during apheresis procedure. Comparing CD62L surface levels in T-cells from the same donor immediately before collecting the apheresis product, and the final apheresis product we found that this process down-regulated CD62L in both CD4+ and CD8+ T cells (p=0.008). Interestingly, when CD62L levels were analysed in days 30 or 60 after engraftment, they recovered to baseline (p=0.008). However, to investigate the relation between CD62L expression and the development of GVHD in the recipient samples after the engraftment, no differences were observed comparing patients with GVHD to those who did not develop the disease. Discussion: Our prospective study indicates that the CCR7+ T-cells from the donor, which include naïve and central memory T-cells, contain the alloreactive cells with a high ability to mediate GVHD (in the case of both migration and activation). Therefore we suggest that the proportion and functional properties of CCR7+CD4+ and CCR7+CD8+ T-cells in the apheresis could act as a predictive biomarker to both acute and chronic GVHD respectively. Importantly, our study precludes that CD62L is lost in the apheresis and therefore it is not a reliable biomarker for the development of GVHD.

Keywords: CCR7, CD62L, GVHD, SCT

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Authors: Jeongyeon Park, Yeo Jun Yoon, Jiyoung Seo, In Seok Moon, Hae Jun Lee, Kiwon Song

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Cold Atmospheric Pressure Plasma (CAPP) is defined as a partially ionized gas with electrically charged particles at room temperature and atmospheric pressure. CAPP generates reactive oxygen species (ROS) and reactive nitrogen species (RNS), and has potential as a new apoptosis-promoting cancer therapy. With an annular type dielectric barrier discharge (DBD) CAPP-generating device combined with a helium (He) gas feeding system, we showed that CAPP selectively induced apoptosis in various cancer cells while it promoted proliferation of the adipose tissue-derived stem cell (ASC). The apoptotic effect of CAPP was highly selective toward p53-mutated cancer cells. The intracellular ROS was mainly responsible for apoptotic cell death in CAPP-treated cancer cells. CAPP induced apoptosis even in doxorubicin-resistant cancer cell lines, demonstrating the feasibility of CAPP as a potent cancer therapy. With the same device and exposure conditions to cancer cells, CAPP stimulated proliferation of the ASC, a kind of mesenchymal stem cell that is capable of self-renewing and differentiating into adipocytes, chondrocytes, osteoblasts and neurons. CAPP-treated ASCs expressed the stem cell markers and differentiated into adipocytes as untreated ASCs. The increase of proliferation by CAPP in ASCs was offset by a NO scavenger but was not affected by ROS scavengers, suggesting that NO generated by CAPP is responsible for the activated proliferation in ASCs. Usually, cancer stem cells are reported to be resistant to known cancer therapies. When we applied CAPP of the same device and exposure conditions to cancer cells to liver cancer stem cells (CSCs) that express CD133 and epithelial cell adhesion molecule (EpCAM) cancer stem cell markers, apoptotic cell death was not examined. Apoptotic cell death of liver CSCs was induced by the CAPP generated from a device with an air-based flatten type DBD. An exposure of liver CSCs to CAPP decreased the viability of liver CSCs to a great extent, suggesting plasma be used as a promising anti-cancer treatment. To validate whether CAPP can be a promising anti-cancer treatment or an adjuvant modality to eliminate remnant tumor in cancer surgery of vestibular schwannoma, we applied CAPP to mouse schwannoma cell line SC4 Nf2 ‑/‑ and human schwannoma cell line HEI-193. A CAPP treatment leads to anti-proliferative effect in both cell lines. We are currently studying the molecular mechanisms of differential physiological effect of CAPP; the proliferation of ASCs and apoptosis of various cancer cells and CSCs.

Keywords: cold atmospheric pressure plasma, apoptosis, proliferation, cancer cells, adult stem cells

Procedia PDF Downloads 259

Authors: Geevika J. Ganegama Arachchi, Steve H. Flint, Lynn McIntyre, Cristina D. Cruz, Beatrice M. Dias-Wanigasekera, Craig Billington, J. Andrew Hudson, Anthony N. Mutukumira

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In seafood processing plants, Listeriamonocytogenes(L. monocytogenes)likely exists in a metabolically stressed state due to the nutrient-deficient environment, processing treatments such as heating, curing, drying, and freezing, and exposure to detergents and disinfectants. Stressed L. monocytogenes cells have been shown to be as pathogenic as unstressed cells. This study investigated lytic efficacy of (LiMN4L, LiMN4p, and LiMN17) which were previouslycharacterized as virulent against physiologically stressed cells of three seafood borne L. monocytogenesstrains (19CO9, 19DO3, and 19EO3).Physiologically compromised cells ofL. monocytogenesstrains were prepared by aging cultures in TrypticaseSoy Broth at 15±1°C for 72 h; heat injuringcultures at 54±1 - 55±1°C for 40 - 60 min;salt-stressing cultures in Milli-Q water were incubated at 25±1°C in darkness for three weeks; and incubating cultures in 9% (w/v) NaCl at 15±1°C for 72 h. Low concentrations of physiologically compromised cells of three L. monocytogenesstrainswere challenged in vitrowith high titre of three phages in separate experiments using Fish Broth medium (aqueous fish extract) at 15 °C in order to mimic the environment of seafood processing plant. Each phage, when present at ≈9 log10 PFU/ml, reduced late exponential phase cells of L. monocytogenes suspended in fish protein broth at ≈2-3 log10 CFU/ml to a non-detectable level (< 10 CFU/ml). Each phage, when present at ≈8.5 log10 PFU/ml, reduced both heat-injured cells present at 2.5-3.6 log10 CFU/ml and starved cells that were showed coccoid shape, present at ≈2-3 log10 CFU/ml to < 10 CFU/ml after 30 min. Phages also reduced salt-stressed cellspresent at ≈3 log10 CFU/ml by > 2 log10. L. monocytogenes (≈8 log10 CFU/ml) were reduced to below the detection limit (1 CFU/ml) by the three successive phage infections over 16 h, indicating that emergence of spontaneous phage resistance was infrequent. The three virulent phages showed high decontamination potential for physiologically stressed L. monocytogenes strains from seafood processing environments.

Keywords: physiologically stressed L. monocytogenes, heat injured, seafood processing environment, virulent phage

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