Search results for: single cell analysis
32045 Morphology Study of Inverted Planar Heterojunction Perovskite Solar Cells in Sequential Deposition
Authors: Asmat Nawaz, Ali Koray Erdinc, Burak Gultekin, Muhammad Tayyib, Ceylan Zafer, Kaiying Wang, M. Nadeem Akram
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In this study, a sequential deposition process is used for the fabrication of PEDOT: PSS based inverted planar perovskite solar cell. A small amount of additive deionized water (DI-H2O) was added into PbI2 + Dimethyl formamide (DMF) precursor solution in order to increase the solubility of PbI2 in DMF, and finally to manipulate the surface morphology of the perovskite films. A morphology transition from needle like structure to hexagonal plates, and then needle-like again has been observed as the DI-H2O was added continuously (0.0 wt% to 3.0wt%). The latter one leads to full surface coverage of the perovskite, which is essential for high performance solar cell.Keywords: charge carrier diffusion lengths, Methylamonium lead iodide, precursor composition, perovskite solar cell, sequential deposition
Procedia PDF Downloads 45232044 Acoustic Radiation Pressure Detaches Myoblast from Culture Substrate by Assistance of Serum-Free Medium
Authors: Yuta Kurashina, Chikahiro Imashiro, Kiyoshi Ohnuma, Kenjiro Takemura
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Research objectives and goals: To realize clinical applications of regenerative medicine, a mass cell culture is highly required. In a conventional cell culture, trypsinization was employed for cell detachment. However, trypsinization causes proliferation decrease due to injury of cell membrane. In order to detach cells using an enzyme-free method, therefore, this study proposes a novel cell detachment method capable of detaching adherent cells using acoustic radiation pressure exposed to the dish by the assistance of serum-free medium with ITS liquid medium supplement. Methods used In order to generate acoustic radiation pressure, a piezoelectric ceramic plate was glued on a glass plate to configure an ultrasonic transducer. The glass plate and a chamber wall compose a chamber in which a culture dish is placed in glycerol. Glycerol transmits acoustic radiation pressure to adhered cells on the culture dish. To excite a resonance vibration of transducer, AC signal with 29-31 kHz (swept) and 150, 300, and 450 V was input to the transducer for 5 min. As a pretreatment to reduce cell adhesivity, serum-free medium with ITS liquid medium supplement was spread to the culture dish before exposed to acoustic radiation pressure. To evaluate the proposed cell detachment method, C2C12 myoblast cells (8.0 × 104 cells) were cultured on a ø35 culture dish for 48 hr, and then the medium was replaced with the serum-free medium with ITS liquid medium supplement for 24 hr. We replaced the medium with phosphate buffered saline and incubated cells for 10 min. After that, cells were exposed to the acoustic radiation pressure for 5 min. We also collected cells by using trypsinization as control. Cells collected by the proposed method and trypsinization were respectively reseeded in ø60 culture dishes and cultured for 24 hr. Then, the number of proliferated cells was counted. Results achieved: By a phase contrast microscope imaging, shrink of lamellipodia was observed before exposed to acoustic radiation pressure, and no cells remained on the culture dish after the exposed of acoustic radiation pressure. This result suggests that serum-free medium with ITS liquid inhibits adhesivity of cells and acoustic radiation pressure detaches cells from the dish. Moreover, the number of proliferated cells 24 hr after collected by the proposed method with 150 and 300 V is the same or more than that by trypsinization, i.e., cells were proliferated 15% higher with the proposed method using acoustic radiation pressure than with the traditional cell collecting method of trypsinization. These results proved that cells were able to be collected by using the appropriate exposure of acoustic radiation pressure. Conclusions: This study proposed a cell detachment method using acoustic radiation pressure by the assistance of serum-free medium. The proposed method provides an enzyme-free cell detachment method so that it may be used in future clinical applications instead of trypsinization.Keywords: acoustic radiation pressure, cell detachment, enzyme free, ultrasonic transducer
Procedia PDF Downloads 25232043 Apoptotic Induction Ability of Harmalol and Its Binding: Biochemical and Biophysical Perspectives
Authors: Kakali Bhadra
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Harmalol administration caused remarkable reduction in proliferation of HepG2 cells with GI50 of 14.2 mM, without showing much cytotoxicity in embryonic liver cell line, WRL-68. Data from circular dichroism and differential scanning calorimetric analysis of harmalol-CT DNA complex shows conformational changes with prominent CD perturbation and stabilization of CT DNA by 8 oC. Binding constant and stoichiometry was also calculated using the above biophysical techniques. Further, dose dependent apoptotic induction ability of harmalol was studied in HepG2 cells using different biochemical assays. Generation of ROS, DNA damage, changes in cellular external and ultramorphology, alteration of membrane, formation of comet tail, decreased mitochondrial membrane potential and a significant increase in Sub Go/G1 population made the cancer cell, HepG2, prone to apoptosis. Up regulation of p53 and caspase 3 further indicated the apoptotic role of harmalol.Keywords: apoptosis, beta carboline alkaloid, comet assay, cytotoxicity, ROS
Procedia PDF Downloads 20532042 Effect of Radioprotectors on DNA Repair Enzyme and Survival of Gamma-Irradiated Cell Division Cycle Mutants of Saccharomyces pombe
Authors: Purva Nemavarkar, Badri Narain Pandey, Jitendra Kumar
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Introduction: The objective was to understand the effect of various radioprotectors on DNA damage repair enzyme and survival in gamma-irradiated wild and cdc mutants of S. pombe (fission yeast) cultured under permissive and restrictive conditions. DNA repair process, as influenced by radioprotectors, was measured by activity of DNA polymerase in the cells. The use of single cell gel electrophoresis assay (SCGE) or Comet Assay to follow gamma-irradiation induced DNA damage and effect of radioprotectors was employed. In addition, studying the effect of caffeine at different concentrations on S-phase of cell cycle was also delineated. Materials and Methods: S. pombe cells grown at permissive temperature (250C) and/or restrictive temperature (360C) were followed by gamma-radiation. Percentage survival and activity of DNA Polymerase (yPol II) were determined after post-irradiation incubation (5 h) with radioprotectors such as Caffeine, Curcumin, Disulphiram, and Ellagic acid (the dose depending on individual D 37 values). The gamma-irradiated yeast cells (with and without the radioprotectors) were spheroplasted by enzyme glusulase and subjected to electrophoresis. Radio-resistant cells were obtained by arresting cells in S-phase using transient treatment of hydroxyurea (HU) and studying the effect of caffeine at different concentrations on S-phase of cell cycle. Results: The mutants of S. pombe showed insignificant difference in survival when grown under permissive conditions. However, growth of these cells under restrictive temperature leads to arrest in specific phases of cell cycle in different cdc mutants (cdc10: G1 arrest, cdc22: early S arrest, cdc17: late S arrest, cdc25: G2 arrest). All the cdc mutants showed decrease in survival after gamma radiation when grown at permissive and restrictive temperatures. Inclusion of the radioprotectors at respective concentrations during post irradiation incubation showed increase in survival of cells. Activity of DNA polymerase enzyme (yPol II) was increased significantly in cdc mutant cells exposed to gamma-radiation. Following SCGE, a linear relationship was observed between doses of irradiation and the tail moments of comets. The radioprotection of the fission yeast by radioprotectors can be seen by the reduced tail moments of the yeast comets. Caffeine also exhibited its radio-protective ability in radio-resistant S-phase cells obtained after HU treatment. Conclusions: The radioprotectors offered notable radioprotection in cdc mutants when added during irradiation. The present study showed activation of DNA damage repair enzyme (yPol II) and an increase in survival after treatment of radioprotectors in gamma irradiated wild type and cdc mutants of S. pombe cells. Results presented here showed feasibility of applying SCGE in fission yeast to follow DNA damage and radioprotection at high doses, which are not feasible with other eukaryotes. Inclusion of caffeine at 1mM concentration to S phase cells offered protection and did not decrease the cell viability. It can be proved that at minimal concentration, caffeine offered marked radioprotection.Keywords: radiation protection, cell cycle, fission yeast, comet assay, s-phase, DNA repair, radioprotectors, caffeine, curcumin, SCGE
Procedia PDF Downloads 10032041 An Ab Initio Study of Delafossite Transparent Conductive Oxides Cu(In, Ga)O2 and Absorbers Films Cu(In, Ga)S2 in Solar-Cell
Authors: Mokdad Sakhri, Youcef Bouhadda
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Thin film chalcopyrite technology is thus nowadays a solid candidate for photovoltaic cells. The currently used window layer for the solar cell Cu(In,Ga)S2 is our interest point in this work. For this purpose, we have performed a first-principles study of structural, electronic and optical properties for both delafossite transparent conductive oxides Cu (In, Ga)O2 and absorbers films Cu(In,Ga)S2. The calculations have been carried out within the local density functional (LDA) and generalized gradient approximations (GGA) combined with the hubbard potential using norm-conserving pseudopotentials and a plane-wave basis with ABINIT code. We have found the energy gap is :1.6, 2.53, 3.6, 3.8 eV for CuInS2, CuGaS2, CuInO2 and CuGaO2 respectively. The results are in good agreement with experimental results.Keywords: ABINIT code, DFT, electronic and optical properties, solar-cell absorbers, delafossite transparent conductive oxides
Procedia PDF Downloads 56232040 Vibration of Gamma Graphyne with an Attached Mass
Authors: Win-Jin Chang, Haw-Long Lee, Yu-Ching Yang
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Atomic finite element simulation is applied to investigate the vibration frequency of a single-layer gamma graphyne with an attached mass for the CCCC, SSSS, CFCF, SFSF boundary conditions using the commercial code ANSYS. The fundamental frequencies of the graphyne sheet are compared with the results of the previous study. The results of the comparison are very good in all considered cases. The attached mass causes a shift in the resonant frequency of the graphyne. The frequencies of the single-layer gamma graphyne with an attached mass for different boundary conditions are obtained, and the order based on the boundary condition is CCCC >SSSS > CFCF> SFSF. The highest frequency shift is obtained when the attached mass is located at the center of the graphyne sheet. This is useful for the design of a highly sensitive graphyne-based mass sensor.Keywords: graphyne, finite element analysis, vibration analysis, frequency shift
Procedia PDF Downloads 20232039 The Physiological Effect of Cold Atmospheric Pressure Plasma on Cancer Cells, Cancer Stem Cells, and Adult Stem Cells
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 27732038 Urinary Exosome miR-30c-5p as a Biomarker for Early-Stage Clear Cell Renal Cell Carcinoma
Authors: Shangqing Song, Bin Xu, Yajun Cheng, Zhong Wang
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miRNAs derived from exosomes exist in a body fluid such as urine were regarded as potential biomarkers for various human cancers diagnosis and prognosis, as mature miRNAs can be steadily preserved by exosomes. However, its potential value in clear cell renal cell carcinoma (ccRCC) diagnosis and prognosis remains unclear. In the present study, differentially expressed miRNAs from urinal exosomes were identified by next-generation sequencing (NGS) technology. The 16 differentially expressed miRNAs were identified between ccRCC patients and healthy donors. To explore the specific diagnosis biomarker of ccRCC, we validated these urinary exosomes from 70 early-stage renal cancer patients, 30 healthy people and other urinary system cancers, including 30 early-stage prostate cancer patients and 30 early-stage bladder cancer patients by qRT-PCR. The results showed that urinary exosome miR-30c-5p could be stably amplified and meanwhile the expression of miR-30c-5p has no significant difference between other urinary system cancers and healthy control, however, expression level of miR-30c-5p in urinary exosomal of ccRCC patients was lower than healthy people and receiver operation characterization (ROC) curve showed that the area under the curve (AUC) values was 0.8192 (95% confidence interval was 0.7388-0.8996, P= 0.0000). In addition, up-regulating miR-30c-5p expression could inhibit renal cell carcinoma cells growth. Lastly, HSP5A was found as a direct target gene of miR-30c-5p. HSP5A depletion reversed the promoting effect of ccRCC growth casued by miR-30c-5p inhibitor, respectively. In conclusion, this study demonstrated that urinary exosomal miR-30c-5p is readily accessible as diagnosis biomarker of early-stage ccRCC, and miR-30c-5p might modulate the expression of HSPA5, which correlated with the progression of ccRCC.Keywords: clear cell renal cell carcinoma, exosome, HSP5A, miR-30c-5p
Procedia PDF Downloads 25632037 Micromorphological Traits and Essential Oil Contents of Valeriana tuberosa L.
Authors: Nada Bezić, Valerija Dunkić, Antonija Markovina, Mirko Rušćić
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Valeriana is a genus of the well-known medicinal plant of Valerianacea family and growing wild in the sub-Mediterranean area. This abstract reports the types and distribution of trichomes and phyto-active composition of the essential oil of the Valeriana tuberosa from mountain Kozjak, near Split, Croatia. Two types of glandular trichomes: peltate (one basal epidermal cell, one short stalk cell and a small head) and capitate trichomes (one basal epidermal cell, one elongated stalk cell) were observed on leaf, using light microscopy. We analyzed the composition of the essential oil of stems and leaves of V. tuberosa species. Water distilled essential oils from aerial parts of investigation plant have been analysed by GC and GC/MS using VF-5ms capillary column. The total yield of oil was 0.2%, based on dry weight of samples. Forty compounds representing 94.1% of the total oil of V. tuberosa. This essential oil was characterized by a high concentration of isovaleric acid (17.2%), geranyl isovalerate (12.2%) and caryophyllene oxide (7.7%). The present study gives additional knowledge about micromorphological traits and secondary metabolites contents on the genus Valeriana.Keywords: essential oil, isovaleric acid, Valeriana tuberosa, Croatia
Procedia PDF Downloads 22532036 Profiling of Apoptotic Protein Expressions after Trabectedin Treatment in Human Prostate Cancer Cell Line PC-3 by Protein Array Technology
Authors: Harika Atmaca, Emir Bozkurt, Latife Merve Oktay, Selim Uzunoglu, Ruchan Uslu, Burçak Karaca
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Microarrays have been developed for highly parallel enzyme-linked immunosorbent assay (ELISA) applications. The most common protein arrays are produced by using multiple monoclonal antibodies, since they are robust molecules which can be easily handled and immobilized by standard procedures without loss of activity. Protein expression profiling with protein array technology allows simultaneous analysis of the protein expression pattern of a large number of proteins. Trabectedin, a tetrahydroisoquinoline alkaloid derived from a Caribbean tunicate, Ecteinascidia turbinata, has been shown to have antitumor effects. Here, we used a novel proteomic approach to explore the mechanism of action of trabectedin in prostate cancer cell line PC-3 by apoptosis antibody microarray. XTT cell proliferation kit and Cell Death Detection Elisa Plus Kit (Roche) was used for measuring cytotoxicity and apoptosis. Human Apoptosis Protein Array (R&D Systems) which consists of 35 apoptosis related proteins was used to assess the omic protein expression pattern. Trabectedin induced cytotoxicity and apoptosis in prostate cancer cells in a time and concentration-dependent manner. The expression levels of the death receptor pathway molecules, TRAIL-R1/DR4, TRAIL R2/DR5, TNF R1/TNFRSF1A, FADD were significantly increased by 4.0-, 21.0-, 4.20- and 11.5-fold by trabectedin treatment in PC-3 cells. Moreover, mitochondrial pathway related pro-apoptotic proteins Bax, Bad, Cytochrome c, and Cleaved Caspase-3 expressions were induced by 2.68-, 2.07-, 2.8-, and 4.5-fold and the expression levels of anti-apoptotic proteins Bcl-2 and Bcl-XL were reduced by 3.5- and 5.2-fold in PC-3 cells. Proteomic (antibody microarray) analysis suggests that the mechanism of action of trabectedin may be exerted via the induction of both intrinsic and extrinsic apoptotic pathways. The antibody microarray platform can be utilised to explore the molecular mechanism of action of novel anticancer agents.Keywords: trabectedin, prostate cancer, omic protein expression profile, apoptosis
Procedia PDF Downloads 43932035 The Relationship Between Social Support, Happiness, Work-Family Conflict and State-Trait Anxiety Among Single Mothers by Choice at Time of Covid-19 Pandemic
Authors: Shamir Balderman Orit, Shamir Michal
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Israel often deals with crisis situations, but most have been characterized as security crises (e.g., war). This is the first time that the Israel has dealt with a health and social emergency as part of a global crisis. The crisis began in January 2020 with the emergence of the novel coronavirus (Covid-19), which was defined as a pandemic (World Health Organization, 2020) and arrived in Israel in early March 2020. This study examined how single mothers by choice (SMBC) experience state anxiety (SA), social support, work–family conflict (WFC), and happiness. This group has not been studied in the context of crises in general or a global crisis. Using a snowball sample, 386 SMBCanswered an online questionnaire. The findings show a negative relationship between income and level of state anxiety. State anxiety was also negatively associated with social support, level of happiness, and WFC. Finally, a stepwise regression analysis indicated that happiness explained 34% of the variance in SA. We also found that most of the women did not turn to formal support agencies such as social workers, other Government Ministries, or municipal welfare. A positive and strong correlations was also found between SA and WFC. The findings of the study reinforce the understanding that although these women made a conscious and informed decision regarding the choice of their family cell, their situation is more complex in the absence of a spouse support. Therefore, this study, as other future studies in the field of SMBC, may contribute to the improvement of their social status and the understanding that they are a unique group. Although SMBC are a growing sector of society in the past few years, there are still special needs and special attention that is needed from the formal and informal supports systems. A comparative study of these two groups and in different countries would shed light on SA among mothers in general, regardless of their relationship status and location. Researchers should expand this study by comparing mothers in relationships and exploring how SMBC coped in other countries. In summary, the findings of the study contribute knowledge on three levels: (a) knowledge about SMBC in general and during crisis situations; (b) examination of social support using tools assessing receipt of assistance and support, some of which were developed for the present study; and (c) insights regarding counseling, accompaniment, and guidance of welfare mechanisms.Keywords: single mothers by choice, state anxiety, social support, happiness, work-family conflict
Procedia PDF Downloads 10032034 Inhibitory Effect of 13-Butoxyberberine Bromide on Metastasis of Skin Cancer A431 Cells
Authors: Phuriwat Laomethakorn, Siritron Samosorn, Ramida Watanapokasin
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Cancer metastasis is the major cause of cancer-related death. Therefore searching for a compound that could inhibit cancer metastasis is necessary. 13-Butoxyberberine bromide is a berberine derivative that has not been reported an anti-metastatic effect on skin cancer cells. This study aimed to investigate the anti-metastatic effect of 13-butoxyberberine bromide on skin cancer A431 cells. The effect of 13-butoxyberberine bromide on A431 cell viability was examined by MTT assay. Suppression of cell migration and invasion in A431 cells were determined by wound healing assay, transwell migration assay, and transwell invasion assay. Metastasis proteins were determined by western blotting. The results demonstrated that 13-butoxyberberine bromide decreased A431 cell viability in a dose-dependent manner. In addition, sub-toxic concentrations of 13-butoxyberberine bromide suppressed cell migration and invasion in A431 cells. In addition, 13-butoxyberberine bromide showed anti-metastatic effects by down-regulated MMP-2 and MMP-9 expression. These findings may be useful in the development of 13-butoxyberberine bromide as an anti-metastatic drug in the future.Keywords: 13-butoxyberberine bromide, metastasis, skin cancer, MMP
Procedia PDF Downloads 9832033 Cascaded Multi-Level Single-Phase Switched Boost Inverter
Authors: Van-Thuan Tran, Minh-Khai Nguyen, Geum-Bae Cho
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Recently, multilevel inverters have become more attractive for researchers due to low total harmonic distortion (THD) in the output voltage and low electromagnetic interference (EMI). This paper proposes a single-phase cascaded H-bridge quasi switched boost inverter (CHB-qSBI) for renewable energy sources applications. The proposed inverter has the advantage over the cascaded H-bridge quasi-Z-source inverter (CHB-qZSI) in reducing two capacitors and two inductors. As a result, cost, weight, and size are reduced. Furthermore, the dc-link voltage of each module is controlled by individual shoot-through duty cycle to get the same values. Therefore, the proposed inverter solves the imbalance problem of dc-link voltage in traditional CHB inverter. This paper shows the operating principles and analysis of the single-phase cascaded H-bridge quasi switched boost inverter. Also, a control strategy for the proposed inverter is shown. Experimental and simulation results are shown to verify the operating principle of the proposed inverter.Keywords: renewable energy sources, cascaded h-bridge inverter, quasi switched boost inverter, quasi z-source inverter, multilevel inverter
Procedia PDF Downloads 32832032 Identification of Individuals in Forensic Situations after Allo-Hematopoietic Stem Cell Transplantation
Authors: Anupuma Raina, Ajay Parkash
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In forensic investigation, DNA analysis helps in the identification of a particular individual under investigation. A set of Short Tandem Repeats loci are widely used for individualization at a molecular level in forensic testing. STRs with tetrameric repeats of DNA are highly polymorphic and widely used for forensic DNA analysis. Identification of an individual became challenging for forensic examiners after Hematopoietic Stem Cell Transplantation. HSCT is a well-accepted and life-saving treatment to treat malignant and nonmalignant diseases. It involves the administration of healthy donor stem cells to replace the patient’s own unhealthy stem cells. A successful HSCT results in complete donor-derived cells in a patient’s hematopoiesis and hence have the capability to change the genetic makeup of the patient. Although an individual who has undergone HSCT and then committed a crime is a very rare situation, but not impossible. Keeping such a situation in mind, various biological samples like blood, buccal swab, and hair follicle were collected and studied after a certain interval of time after HSCT. Blood was collected from both the patient and the donor before the transplant. The DNA profile of both was analyzed using a short tandem repeat kit for autosomal chromosomes. Among all exhibits studied, only hair follicles were found to be the most suitable biological exhibit, as no donor DNA profile was observed for up to 90 days of study.Keywords: chimerism, HSCT, STRs analysis, forensic identification
Procedia PDF Downloads 6132031 New 5’-O- and 6-Substituted Purine Nucleoside Analogs: Synthesis and Cytotoxic Activity on Selected Human Cancer Cell Lines
Authors: Meral Tuncbilek, Duygu Sac, Irem Durmaz, Rengul Cetin Atalay
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Nucleoside analogs are a pharmacologically diverse family that includes cytotoxic compounds, antiviral agents, and immunosuppressive molecules. Purine nucleoside derivatives such as fludarabine, cladribine, and pentostatin are significant drugs used in chemotherapy for the treatment of solid tumors and hematological malignancies. In this study, we synthesized novel purine ribonucleoside analogs containing a 4-(4-substituted phenylsulfonyl) piperazine in the substituent at N6- and O-substituted sulfonyl group at 5’-position as putative cytotoxic agents. The newly obtained compounds were then characterized for their cytotoxicity in human cancer cell lines. The 5’, 6-disubstituted 9-(β-D-ribofuranosyl)purine derivatives (44-67) were readily obtained from commercially available inosine in seven steps in very cost effective synthesis approach. The newly synthesized compounds were first evaluated for their anti-tumor activities against human liver (Huh7), colon (HCT116) and breast (MCF7) carcinoma cell lines. The IC50 values were in micromolar concentrations with 5’, 6-disubstituted purine nucleoside derivatives. Time-dependent IC50 values for each molecule were also calculated in comparison with known cytotoxic agents Camptothecin (CPT), 5-Fluorouracil (5-FU), Cladribine, Pentostatine and Fludarabine. N6-(4-trifluoromethyl phenyl) / N6-(4-bromophenyl) and 5’-O-(4-methoxybenzene sulfonyl) / 5’-O-(benzenesulfonyl) derivatives 54, 64 displayed the best cytotoxic activity with IC50 values of 8.8, 7 µM against MCF7 cell line. The N6-(4-methylphenyl) analog 50 was also very active (IC50= 10.7 μM) against HCT116 cell line. Furthermore, compound 64 had a better cytotoxic activity than the known cell growth inhibitors 5-FU and Fludarabine on Huh7 (1.5 vs 30.7, 29.9 μM for 5-FU and Fludarabine).Keywords: cytotoxic activity, Huh7, HCT116, MCF7, nucleoside, synthesis
Procedia PDF Downloads 23932030 Comfort Evaluation of Summer Knitted Clothes of Tencel and Cotton Fabrics
Authors: Mona Mohamed Shawkt Ragab, Heba Mohamed Darwish
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Context: Comfort properties of garments are crucial for the wearer, and with the increasing demand for cotton fabric, there is a need to explore alternative fabrics that can offer similar or superior comfort properties. This study focuses on comparing the comfort properties of tencel/cotton single jersey fabric and cotton single jersey fabric, with the aim of identifying fabrics that are more suitable for summer clothes. Research Aim: The aim of this study is to evaluate the comfort properties of tencel/cotton single jersey fabric and cotton single jersey fabric, with the goal of identifying fabrics that can serve as alternatives to cotton, considering their comfort properties for summer clothing. Methodology: An experimental, analytical approach was employed in this study. Two circular knitting machines were used to produce the fabrics, one with a 24 inches gauge and the other with a 28 inches gauge. Both fabrics were knitted with three different loop lengths (3.05 mm, 2.9 mm, and 2.6 mm) to obtain loose, medium, and tight fabrics for evaluation. Various comfort properties, including air permeability, water vapor permeability, wickability, and thermal resistance, were measured for both fabric types. Findings: The study found a significant difference in comfort properties between tencel/cotton single jersey fabric and cotton single jersey fabric. Tencel/cotton fabric exhibited higher air permeability, water vapor permeability, and wickability compared to cotton fabric. These findings suggest that tencel fabric is more suitable for summer clothes due to its superior ventilation and absorption properties. Theoretical Importance: This study contributes to the exploration of alternative fabrics to cotton by evaluating their comfort properties. By identifying fabrics that offer better comfort properties than cotton, particularly in terms of water usage, the study provides valuable insights into sustainable fabric choices for the fashion industry. Data Collection and Analysis Procedures: The comfort properties of the fabrics were measured using appropriate testing methods. Paired comparison t-tests were conducted to determine the significant differences between tencel/cotton fabric and cotton fabric in the measured properties. Correlation coefficients were also calculated to examine the relationships between the factors under study. Question Addressed: The study addresses the question of whether tencel/cotton single jersey fabric can serve as an alternative to cotton fabric for summer clothes, considering their comfort properties. Conclusion: The study concludes that tencel/cotton single jersey fabric offers superior comfort properties compared to cotton single jersey fabric, making it a suitable alternative for summer clothes. The findings also highlight the importance of considering fabric properties, such as air permeability, water vapor permeability, and wickability, when selecting materials for garments to enhance wearer comfort. This research contributes to the search for sustainable alternatives to cotton and provides valuable insights for the fashion industry in making informed fabric choices.Keywords: comfort properties, cotton fabric, tencel fabric, single jersey
Procedia PDF Downloads 6832029 Stimulation of Nerve Tissue Differentiation and Development Using Scaffold-Based Cell Culture in Bioreactors
Authors: Simon Grossemy, Peggy P. Y. Chan, Pauline M. Doran
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Nerve tissue engineering is the main field of research aimed at finding an alternative to autografts as a treatment for nerve injuries. Scaffolds are used as a support to enhance nerve regeneration. In order to successfully design novel scaffolds and in vitro cell culture systems, a deep understanding of the factors affecting nerve regeneration processes is needed. Physical and biological parameters associated with the culture environment have been identified as potentially influential in nerve cell differentiation, including electrical stimulation, exposure to extracellular-matrix (ECM) proteins, dynamic medium conditions and co-culture with glial cells. The mechanisms involved in driving the cell to differentiation in the presence of these factors are poorly understood; the complexity of each of them raises the possibility that they may strongly influence each other. Some questions that arise in investigating nerve regeneration include: What are the best protein coatings to promote neural cell attachment? Is the scaffold design suitable for providing all the required factors combined? What is the influence of dynamic stimulation on cell viability and differentiation? In order to study these effects, scaffolds adaptable to bioreactor culture conditions were designed to allow electrical stimulation of cells exposed to ECM proteins, all within a dynamic medium environment. Gold coatings were used to make the surface of viscose rayon microfiber scaffolds (VRMS) conductive, and poly-L-lysine (PLL) and laminin (LN) surface coatings were used to mimic the ECM environment and allow the attachment of rat PC12 neural cells. The robustness of the coatings was analyzed by surface resistivity measurements, scanning electron microscope (SEM) observation and immunocytochemistry. Cell attachment to protein coatings of PLL, LN and PLL+LN was studied using DNA quantification with Hoechst. The double coating of PLL+LN was selected based on high levels of PC12 cell attachment and the reported advantages of laminin for neural differentiation. The underlying gold coatings were shown to be biocompatible using cell proliferation and live/dead staining assays. Coatings exhibiting stable properties over time under dynamic fluid conditions were developed; indeed, cell attachment and the conductive power of the scaffolds were maintained over 2 weeks of bioreactor operation. These scaffolds are promising research tools for understanding complex neural cell behavior. They have been used to investigate major factors in the physical culture environment that affect nerve cell viability and differentiation, including electrical stimulation, bioreactor hydrodynamic conditions, and combinations of these parameters. The cell and tissue differentiation response was evaluated using DNA quantification, immunocytochemistry, RT-qPCR and functional analyses.Keywords: bioreactor, electrical stimulation, nerve differentiation, PC12 cells, scaffold
Procedia PDF Downloads 23632028 Synthesis and Anti-Cancer Evaluation of Uranyle Complexes
Authors: Abdol-Hassan Doulah
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In this research, some of the inorganic complexes of uranyl with N- donor ligands were synthesized. Complexes were characteriezed by FT-IR and UV spectra, ¹HNMR, ¹³CNMR and some physical properties. The uranyl unit (UO2) is composed of a center of uranium atom with the charge (+6) and two oxygen atom by forming two U=O double bonds. The structure is linear (O=U=O, 180) and usually stable. So other ligands often coordinate to the U atom in the plane perpendicularly to the O=U=O axis. The antitumor activity of some of ligand and their complexes against a panel of human tumor cell lines (HT29: Haman colon adenocarcinoma cell line T47D: human breast adenocarcinoma cell line) were determined by MTT(3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) assay. These data suggest that some of these compounds provide good models for the further design of potent antitumor compounds.Keywords: inorganic, uranyl complex-donor ligands, Schiff bases, anticancer activity
Procedia PDF Downloads 45032027 Gall Bladder Polyp Identified as Solitary RCC Metastasis 4 Years after Nephrectomy: An Unusual Case Report
Authors: Gerard Bray, Arya Bahadori, Sachinka Ranasinghe
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Renal cell carcinoma (RCC) is among the top 10 most common cancers worldwide, where metastatic disease carries a poor prognosis. Herein, we present a 74-year-old male presenting with asymptomatic solitary metachronous metastasis to the gall bladder 4 years following nephrectomy for clear cell RCC. Solitary RCC metastasis to the gall bladder following nephrectomy is rarely reported in the literature and brings with it a clinical conundrum of whether surgical resection or systemic therapy should be utilized. In this case, surgical excision with cholecystectomy was employed without systemic therapy. We, therefore, contribute a rare and interesting case that highlights that metastasectomy of a solitary metastasis can improve survival according to current literature.Keywords: renal cell carcinoma, gall bladder metastasis, solitary metastasectomy, metachronous
Procedia PDF Downloads 16532026 Constitutive Modeling of Different Types of Concrete under Uniaxial Compression
Authors: Mostafa Jafarian Abyaneh, Khashayar Jafari, Vahab Toufigh
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The cost of experiments on different types of concrete has raised the demand for prediction of their behavior with numerical analysis. In this research, an advanced numerical model has been presented to predict the complete elastic-plastic behavior of polymer concrete (PC), high-strength concrete (HSC), high performance concrete (HPC) along with different steel fiber contents under uniaxial compression. The accuracy of the numerical response was satisfactory as compared to other conventional simple models such as Mohr-Coulomb and Drucker-Prager. In order to predict the complete elastic-plastic behavior of specimens including softening behavior, disturbed state concept (DSC) was implemented by nonlinear finite element analysis (NFEA) and hierarchical single surface (HISS) failure criterion, which is a failure surface without any singularity.Keywords: disturbed state concept (DSC), hierarchical single surface (HISS) failure criterion, high performance concrete (HPC), high-strength concrete (HSC), nonlinear finite element analysis (NFEA), polymer concrete (PC), steel fibers, uniaxial compression test
Procedia PDF Downloads 30832025 3D Microscopy, Image Processing, and Analysis of Lymphangiogenesis in Biological Models
Authors: Thomas Louis, Irina Primac, Florent Morfoisse, Tania Durre, Silvia Blacher, Agnes Noel
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In vitro and in vivo lymphangiogenesis assays are essential for the identification of potential lymphangiogenic agents and the screening of pharmacological inhibitors. In the present study, we analyse three biological models: in vitro lymphatic endothelial cell spheroids, in vivo ear sponge assay, and in vivo lymph node colonisation by tumour cells. These assays provide suitable 3D models to test pro- and anti-lymphangiogenic factors or drugs. 3D images were acquired by confocal laser scanning and light sheet fluorescence microscopy. Virtual scan microscopy followed by 3D reconstruction by image aligning methods was also used to obtain 3D images of whole large sponge and ganglion samples. 3D reconstruction, image segmentation, skeletonisation, and other image processing algorithms are described. Fixed and time-lapse imaging techniques are used to analyse lymphatic endothelial cell spheroids behaviour. The study of cell spatial distribution in spheroid models enables to detect interactions between cells and to identify invasion hierarchy and guidance patterns. Global measurements such as volume, length, and density of lymphatic vessels are measured in both in vivo models. Branching density and tortuosity evaluation are also proposed to determine structure complexity. Those properties combined with vessel spatial distribution are evaluated in order to determine lymphangiogenesis extent. Lymphatic endothelial cell invasion and lymphangiogenesis were evaluated under various experimental conditions. The comparison of these conditions enables to identify lymphangiogenic agents and to better comprehend their roles in the lymphangiogenesis process. The proposed methodology is validated by its application on the three presented models.Keywords: 3D image segmentation, 3D image skeletonisation, cell invasion, confocal microscopy, ear sponges, light sheet microscopy, lymph nodes, lymphangiogenesis, spheroids
Procedia PDF Downloads 36632024 Changes in Global DNA Methylation and DNA Damage in Two Tumor Cell Lines Treated with Silver and Gold Nanoparticles
Authors: Marcin Kruszewski, Barbara Sochanowicz, Sylwia Męczyńska-Wielgosz, Maria Wojewódzka, Lucyna Kapka-Skrzypczak
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Metallic NPs are widely used in a number of applications in industry, science and medicine. Among metallic NPs foreseen to be widely used in medicine are gold nanoparticles (AuNPs) due to their low toxicity, and silver NPs (AgNPs) due to their strong antimicrobial activity. In this study, we compared an effect of AgNPs and gold NPs (AuNPs) on the formation of DNA damage and global DNA methylation and in A2780 and 4T1 cell lines, widely used models of human ovarian carcinoma and murine mammary carcinoma, respectively. The cells were treated with AgNPs coated with citrate (AgNPs(cit) or PEG (AgNPs(PEG), or AuNPs. A global DNA methylation was investigated with ELISA, whereas the formation of DNA damage was investigated by a comet +/- FPG. AgNPs decreased global DNA methylation and increased the formation of DNA lesions in both cell lines. The effect was dependent on the type of NPs used, it's coating, and cell line used. In conclusion, the epigenetic and genotoxic effects of NPs strongly depends on NP nature and cellular context. Epigenetic changes observed upon the action of AgNPs may play a crucial role in NPs-induced changes in protein expression.Keywords: DNA damage, gold nanoparticles, methylation, silver nanoparticles
Procedia PDF Downloads 12932023 A Fluid-Walled Microfluidic Device for Cell Migration Studies
Authors: Cyril Deroy, Agata Rumianek, David R. Greaves, Peter R. Cook, Edmond J. Walsh
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Various microfluidic platforms have been developed in the past couple of decades offering experimental methods for the study of cell migration; however, their implementation in the laboratory has remained limited. Some reasons cited for the lack of uptake include the technical complexity of the devices, high failure rate associated with gas-bubbles, biocompatibility concerns with the use of polydimethylsiloxane (PDMS) and equipment/time/expertise requirements for operation and manufacture. As sample handling remains challenging due to the closed format of microfluidic devices, open microfluidic systems have been developed offering versatility and simplicity of use. Rather than confining fluids by solid walls, samples can be accessed directly over the open platform, by removing at least one of the solid boundaries, such as the cover. In this paper, a method for the fabrication of open fluid-walled microfluidic circuits for cell migration studies is introduced, where only materials commonly used by the life-science community are required; tissue culture dishes and cell media. The simplicity of the method, and ability to retrieve cells of interest are two key features of the method. Both passive and active flow-devices can be created in this way. To demonstrate the versatility of the method a cell migration assay is performed, which requires fabricating circuits for establishing chemical gradients, loading cells and incubating, creating chemical gradients, real time imaging of cell migration and finally retrieval of cells. The open architecture has high fidelity as it eliminates air bubble related failures and enables the precise control of gradients. The ability to fabricate custom microfluidic designs in minutes should make this method suitable for use in a wide range of cell migration studies.Keywords: chemotaxis, fluid walls, gradient generation, open microfluidics
Procedia PDF Downloads 14532022 Chemotactic Behaviour of Human Mesenchymal Stem Cells in Response to Silicate Substituted Hydroxyapatite
Authors: Dinara Ikramova, Karin A. Hing, Simon C. F. Rawlinson
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Silicate-substituted hydroxyapatite (SiHA) has been shown to enhance bone regeneration in vivo compared with phase pure stoichiometric hydroxyapatite. Evidence suggests that substrate chemistry dependent formation of a permissive protein layer on the surface of synthetic bone graft substitute materials is key for bioactivity and cell attachment. However, little information is available on whether the substrate chemistry may affect cell migration and recruitment. The aim of this study is to investigate whether or not human Mesenchymal Stem Cells (hMSCs) exhibit a chemotactic response to SiHA porous granules and if it can be linked to either the ion exchange or protein sequestering and enrichment on the surface of the material. 150mg of SiHA granules with 80% total porosity and 20% strut porosity were incubated in 1ml of either Serum Free Media (SFM) or 10% Serum Containing Media (SCM) under static cell culture conditions (37°C, 5% CO2) in absence of cells. Protein sequestering and exchange of calcium, phosphate and silicate ions were analysed at 0.5, 1, 2, 4, 8, 16 and 24 hours with n=12 per time point. Migration of hMSCs in the presence of 150mg of SiHA granules was assessed over 24 hours using a modified transwell migration system in either SFM or SCM (n=6) with 30% serum containing media acting as a positive control. At 24 hours protein sequestering and ionic exchange were analysed, and the number of cells was quantified using a high throughput confocal microscope (IN Cell Analyser 6000). In acellular condition, both calcium and phosphate ion concentrations in media showed a decrease at 24 hours which was greater in SFM than in SCM. This suggests possible formation and precipitation of a bone like apatite on the surface of SiHA. Reduction in this activity observed in SCM indicates that the presence of serum proteins is interfering with the ion exchange at the material and media interface. Adsorbed protein levels showed fluctuation over time followed by sharp decrease at 24 hours, suggesting a possible protein rearrangement on the surface of the material. The ion analysis performed on SFM and SCM after 24-hour incubation with cells in the presence of granules showed a greater reduction in phosphate concentration in both SFM and SCM compared to phosphate levels in acellular condition. Silicate concentration in SCM increased from 1.6mM (absence of cells) to 5.1mM (presence of cells). This indicates that the cells are promoting the uptake of phosphate and release of silicate ions. No significant change was seen in levels of adsorbed proteins in the presence and absence of cells. Further analysis is required to determine whether the species of these proteins change over time. The analysis of cell migration after 24-hour incubation showed more cells migrating towards the granules, 12.7% in SFM and 8.3% in SCM, than in positive control, 4.5% in SFM and 3.6% in SCM respectively. These results suggest that SiHA has a chemotactic activity independent of serum proteins. A property which has not previously been demonstrated for a synthetic bone graft material.Keywords: cell migration, hMSCs, SiHA, transwell migration system
Procedia PDF Downloads 12732021 3D Electrode Carrier and its Implications on Retinal Implants
Authors: Diego Luján Villarreal
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Retinal prosthetic devices aim to repair some vision in visual impairment patients by stimulating electrically neural cells in the visual system. In this study, the 3D linear electrode carrier is presented. A simulation framework was developed by placing the 3D carrier 1 mm away from the fovea center at the highest-density cell. Cell stimulation is verified in COMSOL Multiphysics by developing a 3D computational model which includes the relevant retinal interface elements and dynamics of the voltage-gated ionic channels. Current distribution resulting from low threshold amplitudes produces a small volume equivalent to the volume confined by individual cells at the highest-density cell using small-sized electrodes. Delicate retinal tissue is protected by excessive charge densityKeywords: retinal prosthetic devices, visual devices, retinal implants., visual prosthetic devices
Procedia PDF Downloads 9732020 Chaotic Motion of Single-Walled Carbon Nanotube Subject to Damping Effect
Authors: Tai-Ping Chang
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In the present study, the effects on chaotic motion of single-walled carbon nanotube (SWCNT) due to the linear and nonlinear damping are investigated. By using the Hamilton’s principle, the nonlinear governing equation of the single-walled carbon nanotube embedded in a matrix is derived. The Galerkin’s method is adopted to simplify the integro-partial differential equation into a nonlinear dimensionless governing equation for the SWCNT, which turns out to be a forced Duffing equation. The variations of the Lyapunov exponents of the SWCNT with damping and harmonic forcing amplitudes are investigated. Based on the computations of the top Lyapunov exponent, it is concluded that the chaotic motion of the SWCNT occurs when the amplitude of the periodic excitation exceeds certain value, besides, the chaotic motion of the SWCNT occurs with small linear damping and tiny nonlinear damping.Keywords: chaotic motion, damping, Lyapunov exponents, single-walled carbon nanotube
Procedia PDF Downloads 31532019 Numerical Analysis on Triceratops Restraining System: Failure Conditions of Tethers
Authors: Srinivasan Chandrasekaran, Manda Hari Venkata Ramachandra Rao
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Increase in the oil and gas exploration in ultra deep-water demands an adaptive structural form of the platform. Triceratops has superior motion characteristics compared to that of the Tension Leg Platform and Single Point Anchor Reservoir platforms, which is well established in the literature. Buoyant legs that support the deck are position-restrained to the sea bed using tethers with high axial pretension. Environmental forces that act on the platform induce dynamic tension variations in the tethers, causing the failure of tethers. The present study investigates the dynamic response behavior of the restraining system of the platform under the failure of a single tether of each buoyant leg in high sea states. Using the rain-flow counting algorithm and the Goodman diagram, fatigue damage caused to the tethers is estimated, and the fatigue life is predicted. Results shows that under failure conditions, the fatigue life of the remaining tethers is quite alarmingly low.Keywords: fatigue life, pm spectrum, rain flow counting, triceratops, failure analysis
Procedia PDF Downloads 12832018 Model Evaluation of Nanosecond, High-Intensity Electric Pulses Induced Cellular Apoptosis
Authors: Jiahui Song, Ravindra Joshi
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High-intensity, nanosecond, pulsed electric fields have been shown to be useful non-thermal tools capable of producing a variety of specific cellular responses. While reversible and temporary changes are often desired based on electromanipulation, irreversible effects can also be important objectives. These include elimination of tumor cells and bacterial decontamination. A simple model-based rate-equation treatment of the various cellular biochemical processes was used to qualitatively predict the pulse number-dependent caspase activation and cell survival trends. The model incorporated the caspase-8 associated extrinsic pathway, the delay inherent in its activation, cytochrome c release, and the internal feedback mechanism between caspase-3 and Bid. Results were roughly in keeping with the experimental cell-survival data. A pulse-number threshold was predicted followed by a near-exponential fall-off. The intrinsic pathway was shown to be much weaker as compared to the extrinsic mechanism for electric pulse induced cell apoptosis. Also, delays of about an hour are predicted for detectable molecular concentration increases following electrical pulsing.Keywords: apoptosis, cell survival, model, pathway
Procedia PDF Downloads 23332017 Quaternized PPO/PSF Anion Exchange Membranes Doped with ZnO-Nanoparticles for Fuel Cell Application
Authors: P. F. Msomi, P. T. Nonjola, P. G. Ndungu, J. Ramontja
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In view of the projected global energy demand and increasing levels of greenhouse gases and pollutants issues have inspired an intense search for alternative new energy technologies, which will provide clean, low cost and environmentally friendly solutions to meet the end user requirements. Alkaline anion exchange membrane fuel cells (AAEMFC) have been recognized as ideal candidates for the generation of such clean energy for future stationary and mobile applications due to their many advantages. The key component of the AAEMFC is the anion exchange membrane (AEM). In this report, a series of quaternized poly (2.6 dimethyl – 1.4 phenylene oxide)/ polysulfone (QPPO/PSF) blend anionic exchange membranes (AEM) were successfully fabricated and characterized for alkaline fuel cell application. Zinc Oxide (ZnO) nanoparticles were introduced in the polymer matrix to enhance the intrinsic properties of the AEM. The characteristic properties of the QPPO/PSF and QPPO/PSF-ZnO blend membrane were investigated with X-ray diffraction (XRD), thermogravimetric analysis (TGA) scanning electron microscope (SEM) and contact angle (CA). To confirm successful quaternisation, FT-IR spectroscopy and proton nuclear magnetic resonance (1H NMR) were used. Other properties such as ion exchange capacity (IEC), water uptake, contact angle and ion conductivity (IC) were also undertaken to check if the prepared nanocomposite materials are suitable for fuel cell application. The membrane intrinsic properties were found to be enhanced by the addition of ZnO nanoparticles. The addition of ZnO nanoparticles resulted to a highest IEC of 3.72 mmol/g and a 30-fold IC increase of the nanocomposite due to its lower methanol permeability. The above results indicate that QPPO/PSF-ZnO is a good candidate for AAEMFC application.Keywords: anion exchange membrane, fuel cell, zinc oxide nanoparticle, nanocomposite
Procedia PDF Downloads 41632016 Autophagy Regulates Human Hepatocellular Carcinoma Tumorigenesis through Selective Degradation of Cyclin D1
Authors: Shan-Ying Wu, Sheng-Hui Lan, Xi-Zhang Lin, Ih-Jen Su, Ting-Fen Tsai, Chia-Jui Yen, Tsung-Hsueh Lu, Fu-Wen Liang, Huey-Jen Su, Chun-Li Su, Hsiao-Sheng Liu
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In hepatocelluar carcinoma (HCC), dysregulated expression of cyclin D1 and impaired autophagy has been reported separately. However, the relationship between them has not been explored. In this study, we demonstrated that autophagy was inversely correlated with cyclin D1 expression in 147 paired HCC patient specimens. HCC specimen with highly expression of cyclin D1 shows correlation with poor overall survival rate. Furthermore, induction of autophagy by amiodarone (antiarrhythmic drug) in Hep 3B cells, cyclin D1 was recruited into autophagosomes demonstrated by immune-gold labeling of cyclin D1 after extraction of autophagosomes. We further demonstrated that autophagy suppresses Hep 3B cell proliferation, and further analysis revealed that cell cycle was arrested at G1 phase. The interaction between LC3 (maker of autophagy) and cyclin D1 was increased after autophagy induction. In addition, ubiquitinated-cyclin D1 was also increased after autophagy induction, which is selectively degraded by autophagosome through binding with SQSTM1/p62 (an adaptor protein). In vivo study showed that amiodarone induced autophagy suppresses liver tumor formation in xenograft mouse and orthotopic rat model through decreasing cyclin D1 expression and inhibition of cell proliferation. Altogether, we reveal a novel mechanism that ubiquitinated cyclin D1 degraded by autophagic pathway by p62 and amiodarone is a promising drug for targeting cyclin D1 in liver cancer therapy.Keywords: autophagy, cyclin D1, hepatocellular carcinoma, amiodarone
Procedia PDF Downloads 291