Search results for: immobilised cell
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3574

Search results for: immobilised cell

604 Drought Alters the Expression of a Candidate Zea Mays P-Coumarate 3-Hydroxylase Gene and Caffeic Acid Biosynthesis

Authors: Zintle Kolo, Ndiko Ludidi

Abstract:

The enzymatic activity of p-coumarate 3-hydroxylase (C3H) synthesize caffeic acid from p-coumaric acid. We recently showed that exogenously applied caffeic acid confers salinity tolerance in soybean (Glycine max) by inducing antioxidant enzymatic activity to promote enhanced scavenging or reactive oxygen species, thus limiting salinity-induced oxidative stress. Recent evidence also establishes that pre-treatment of plants with exogenously supplied caffeic acid improves plant tolerance to osmotic stress by improving plant antioxidant capacity and enhancing biosynthesis of compatible solutes. We aimed to identify a C3H in maize (Zea mays) and evaluate the effect of drought on the spatial and temporal expression of the gene encoding the candidate maize C3H (ZmC3H). Primary sequence analysis shows that ZmC3H shares 71% identity with an Arabidopsis thaliana C3H that is implicated in the control of Arabidopsis cell expansion, growth, and responses to stress. In silico ZmC3H promoter analysis reveals the presence of cis-acting elements that interact with transcription factors implicated in plant responses to drought. Spatial expression analysis by semi-quantitative RT-PCR shows that ZmC3H is expressed in both leaves and roots under normal conditions. However, drought represses the expression of ZmC3H in leaves whereas it up-regulates its expression in roots. These changes in ZmC3H expression correlate with the changes in the content of caffeic acid in maize in response to drought. We illustrate the implications of these changes in the expression of the gene in relation to maize responses to drought and discuss the potential of regulating caffeic acid biosynthesis towards genetic improvement of maize tolerance to drought stress. These findings have implications for food security because of the potential of the implications of the study for drought tolerance in maize.

Keywords: caffeic acid, drought-responsive expression, maize drought tolerance, p-coumarate 3-hydroxylase

Procedia PDF Downloads 442
603 Seroprevalence of Toxoplasmosis among Hemato-Oncology Patients in Tertiary Hospital of East Cost Malaysia

Authors: Aisha Khodijah Kholib Jati, Suharni Mohamad, Azlan Husin, Wan Suriana Wan Ab Rahman

Abstract:

Introduction: Toxoplasmosis is caused by an obligate intracellular parasite, Toxoplasma gondii (T. gondii). It is commonly asymptomatic in normal individual, but it can be fatal to immunocompromised patients as it can lead to severe complications such as encephalitis, chorioetinitis and myocarditis. Objective: The aim of the study was to determine the seroprevalence of toxoplasmosis and its association with socio-demographic and behavioral characteristics among hemato-oncology patients in Hospital USM. Methods: In this cross-sectional study, 56 hemato-oncology patients were screened for immunoglobulin M (IgM) antibodies, immunoglobulin G (IgG) antibodies, and IgG avidity of T. gondii by using ELISA Kit (BioRad, USA). For anti-T. gondii IgG antibody, titer ≥ 9 IU/ml was considered as recent infection, while for IgM, ratio ≥ 1.00 was considered as reactive for the anti-T. gondii IgM antibodies. Low avidity index is considered as recent infection within 20 weeks while high avidity considered as past infection. T. gondii exposure, socio-demographic and behavioral characteristics was assessed by a questionnaire and interview. Results: A total of 28 (50.0%) hemato-oncology patients were seropositive for T. gondii antibodies. Out of that total, 27 (48.21%) patients were IgG+/IgM- and one patient (1.79%) was IgG+/IgM+ with high avidity index. Univariate analysis showed that age, gender, ethnicity, marital status, educational level, employment status, stem cell transplant, blood transfusion, close contact with cats, water supply, and consumption of undercooked meat were not significantly associated with Toxoplasma seropositivity rate. Discussion: The seropositivity rate of IgG anti-T. gondii was high among hemato-oncology patients in Hospital USM. With impaired immune system, these patients might have a severe consequence if the infection reactivated. Therefore, screening for anti-T. gondii may be considered in the future. Moreover, health programme towards healthy food and good hygiene practice need to be implemented.

Keywords: immunocompromised, seroprevalence, socio-demographic, toxoplasmosis

Procedia PDF Downloads 132
602 Clinical Efficacy of Nivolumab and Ipilimumab Combination Therapy for the Treatment of Advanced Melanoma: A Systematic Review and Meta-Analysis of Clinical Trials

Authors: Zhipeng Yan, Janice Wing-Tung Kwong, Ching-Lung Lai

Abstract:

Background: Advanced melanoma accounts for the majority of skin cancer death due to its poor prognosis. Nivolumab and ipilimumab are monoclonal antibodies targeting programmed cell death protein 1 (PD-1) and cytotoxic T-lymphocytes antigen 4 (CTLA-4). Nivolumab and ipilimumab combination therapy has been proven to be effective for advanced melanoma. This systematic review and meta-analysis are to evaluate its clinical efficacy and adverse events. Method: A systematic search was done on databases (Pubmed, Embase, Medline, Cochrane) on 21 June 2020. Search keywords were nivolumab, ipilimumab, melanoma, and randomised controlled trials. Clinical trials fulfilling the inclusion criteria were selected to evaluate the efficacy of combination therapy in terms of prolongation of progression-free survival (PFS), overall survival (OS), and objective response rate (ORR). The odd ratios and distributions of grade 3 or above adverse events were documented. Subgroup analysis was performed based on PD-L1 expression-status and BRAF-mutation status. Results: Compared with nivolumab monotherapy, the hazard ratios of PFS, OS and odd ratio of ORR in combination therapy were 0.64 (95% CI, 0.48-0.85; p=0.002), 0.84 (95% CI, 0.74-0.95; p=0.007) and 1.76 (95% CI, 1.51-2.06; p < 0.001), respectively. Compared with ipilimumab monotherapy, the hazard ratios of PFS, OS and odd ratio of ORR were 0.46 (95% CI, 0.37-0.57; p < 0.001), 0.54 (95% CI, 0.48-0.61; p < 0.001) and 6.18 (95% CI, 5.19-7.36; p < 0.001), respectively. In combination therapy, the odds ratios of grade 3 or above adverse events were 4.71 (95% CI, 3.57-6.22; p < 0.001) compared with nivolumab monotherapy, and 3.44 (95% CI, 2.49-4.74; p < 0.001) compared with ipilimumab monotherapy, respectively. High PD-L1 expression level and BRAF mutation were associated with better clinical outcomes in patients receiving combination therapy. Conclusion: Combination therapy is effective for the treatment of advanced melanoma. Adverse events were common but manageable. Better clinical outcomes were observed in patients with high PD-L1 expression levels and positive BRAF-mutation.

Keywords: nivolumab, ipilimumab, advanced melanoma, systematic review, meta-analysis

Procedia PDF Downloads 111
601 Microglia Activity and Induction of Mechanical Allodynia after Mincle Receptor Ligand Injection in Rat Spinal Cord

Authors: Jihoon Yang, Jeong II Choi

Abstract:

Mincle is expressed in macrophages and is members of immunoreceptors induced after exposure to various stimuli and stresses. Mincle receptor activation promotes the production of these substances by increasing the transcription of inflammatory cytokines and chemokines. Cytokines, which play an important role in the initiation and maintenance of such inflammatory pain diseases, have a significant effect on sensory neurons in addition to their enhancement and inhibitory effects on immune and inflammatory cells as mediators of cell interaction. Glial cells in the central nervous system play a critical role in development and maintenance of chronic pain states. Microglia are tissue-resident macrophages in the central nervous system, and belong to a group of mononuclear phagocytes. In the central nervous system, mincle receptor is present in neurons and glial cells of the brain.This study was performed to identify the Mincle receptor in the spinal cord and to investigate the effect of Mincle receptor activation on nociception and the changes of microglia. Materials and Methods: C-type lectins(Mincle) was identified in spinal cord of Male Sprague–Dawley rats. Then, mincle receptor ligand (TDB), via an intrathecal catheter. Mechanical allodynia was measured using von Frey test to evaluate the effect of intrathecal injection of TDB. Result: The present investigation shows that the intrathecal administration of TDB in the rat produces a reliable and quantifiable mechanical hyperalgesia. In addition, The mechanical hyperalgesia after TDB injection gradually developed over time and remained until 10 days. Mincle receptor is identified in the spinal cord, mainly expressed in neuronal cells, but not in microglia or astrocyte. These results suggest that activation of mincle receptor pathway in neurons plays an important role in inducing activation of microglia and inducing mechanical allodynia.

Keywords: mincle, spinal cord, pain, microglia

Procedia PDF Downloads 138
600 Poly-ε-Caprolactone Nanofibers with Synthetic Growth Factor Enriched Liposomes as Controlled Drug Delivery System

Authors: Vera Sovkova, Andrea Mickova, Matej Buzgo, Karolina Vocetkova, Eva Filova, Evzen Amler

Abstract:

PCL (poly-ε-caprolactone) nanofibrous scaffolds with adhered liposomes were prepared and tested as a possible drug delivery system for various synthetic growth factors. TGFβ, bFGF, and IGF-I have been shown to increase hMSC (human mesenchymal stem cells) proliferation and to induce hMSC differentiation. Functionalized PCL nanofibers were prepared with synthetic growth factors encapsulated in liposomes adhered to them in three different concentrations. Other samples contained PCL nanofibers with adhered, free synthetic growth factors. The synthetic growth factors free medium served as a control. The interaction of liposomes with the PCL nanofibers was visualized by SEM, and the release kinetics were determined by ELISA testing. The potential of liposomes, immobilized on the biodegradable scaffolds, as a delivery system for synthetic growth factors, and as a suitable system for MSCs adhesion, proliferation and differentiation in vitro was evaluated by MTS assay, dsDNA amount determination, confocal microscopy, flow cytometry and real-time PCR. The results showed that the growth factors adhered to the PCL nanofibers stimulated cell proliferation mainly up to day 11 and that subsequently their effect was lower. By contrast, the release of the lowest concentration of growth factors from liposomes resulted in gradual proliferation of MSCs throughout the experiment. Moreover, liposomes, as well as free growth factors, stimulated type II collagen production, which was confirmed by immunohistochemical staining using monoclonal antibody against type II collagen. The results of this study indicate that growth factors enriched liposomes adhered to surface of PCL nanofibers could be useful as a drug delivery instrument for application in short timescales, be combined with nanofiber scaffolds to promote local and persistent delivery while mimicking the local microenvironment. This work was supported by project LO1508 from the Ministry of Education, Youth and Sports of the Czech Republic

Keywords: drug delivery, growth factors, hMSC, liposomes, nanofibres

Procedia PDF Downloads 264
599 Contour Defects of Face with Hyperpigmentation

Authors: Afzaal Bashir, Sunaina Afzaal

Abstract:

Background: Facial contour deformities associated with pigmentary changes are of major concern for plastic surgeons, both being important and difficult in treating such issues. No definite ideal treatment option is available to simultaneously address both the contour defect as well as related pigmentation. Objectives: The aim of the current study is to compare the long-term effects of conventional adipose tissue grafting and ex-vivo expanded Mesenchymal Stem Cells enriched adipose tissue grafting for the treatment of contour deformities with pigmentary changes on the face. Material and Methods: In this study, eighty (80) patients with contour deformities of the face with hyperpigmentation were recruited after informed consent. Two techniques i.e., conventional fat grafting (C-FG) and fat grafts enriched with expanded adipose stem cells (FG-ASCs), were used to address the pigmentation. Both techniques were explained to patients, and enrolled patients were divided into two groups i.e., C-FG and FG-ASCS, per patients’ choice and satisfaction. Patients of the FG-ASCs group were treated with fat grafts enriched with expanded adipose stem cells, while patients of the C-FGs group were treated with conventional fat grafting. Patients were followed for 12 months, and improvement in face pigmentation was assessed clinically as well as measured objectively. Patient satisfaction was also documented as highly satisfied, satisfied, and unsatisfied. Results: Mean age of patients was 24.42(±4.49), and 66 patients were females. The forehead was involved in 61.20% of cases, the cheek in 21.20% of cases, the chin in 11.20% of cases, and the nose in 6.20% of cases. In the GF-ASCs group, the integrated color density (ICD) was decreased (1.08×10⁶ ±4.64×10⁵) as compared to the C-FG group (2.80×10⁵±1.69×10⁵). Patients treated with fat grafts enriched with expanded adipose stem cells were significantly more satisfied as compared to patients treated with conventional fat grafting only. Conclusion: Mesenchymal stem cell-enriched autologous fat grafting is a preferred option for improving the contour deformities related to increased pigmentation of face skin.

Keywords: hyperpigmentation, color density, enriched adipose tissue graft, fat grafting, contour deformities, Image J

Procedia PDF Downloads 72
598 Development of Low Calorie Jelly with Increased Content of Natural Compounds from Superfoods with No Added Sugar

Authors: Liana C. Salanță, Maria Tofană, Carmen R. Pop, Vlad Mureșan

Abstract:

The landscape of functional food is expanding very fast, due to the consumer interest for healthy natural products. Consumers nowadays demand healthy products that impart phytonutrients to encourage good health and well-being, prevent diseases, without sacrificing taste and texture. Candies are foodstuffs appreciated by all category of consumers. They are available in a range variety of forms (jellies, marshmallows, caramels, lollipops, etc.). Jelly is characterized by a gummy and chewy texture typically conferred by a hydrocolloid (gelatin, pectin). The purpose of this research was to obtain hypocaloric jelly (no added sugar) enriched with protein powder from acai, chia seeds and hemp, which are considered superfood. Peach and raspberry juice were used for obtaining functional jelly, due to the specific flavour, natural carbohydrate, natural pigments and vitamins (C, B1, PP, etc). Instead of classic hydrocolloids used in Romania for the industry of jelly, agar-agar was used in this study, due to its properties. Agar-agar is able to form gels in the aqueous medium, stronger than other gel-forming agents. High sugar concentrations or an acid environment (as is necessary with pectins) are not needed. In addition to its gelation properties, Agar-agar is considered to have important nutritional benefits, high content of fibre and has low calories. Six prototypes of jellies were obtained and evaluated by physicochemical, microbiological and sensorial analysis. For the textural profile analysis, the Brookfield CT3 Texture Analyzer, equipped with a 10kg load cell, was used. The results revealed that hypocaloric jelly can serve as a good source of bioactive compounds in the diet. The jelly is a convenient way of delivering potential health benefits of protein powder and agar-agar to a wide range of consumers.

Keywords: agar-agar, functional food, hypocaloric jelly, superfoods

Procedia PDF Downloads 106
597 Monitoring the Thin Film Formation of Carrageenan and PNIPAm Microgels

Authors: Selim Kara, Ertan Arda, Fahrettin Dolastir, Önder Pekcan

Abstract:

Biomaterials and thin film coatings play a fundamental role in medical, food and pharmaceutical industries. Carrageenan is a linear sulfated polysaccharide extracted from algae and seaweeds. To date, such biomaterials have been used in many smart drug delivery systems due to their biocompatibility and antimicrobial activity properties. Poly (N-isopropylacrylamide) (PNIPAm) gels and copolymers have also been used in medical applications. PNIPAm shows lower critical solution temperature (LCST) property at about 32-34 °C which is very close to the human body temperature. Below and above the LCST point, PNIPAm gels exhibit distinct phase transitions between swollen and collapsed states. A special class of gels are microgels which can react to environmental changes significantly faster than microgels due to their small sizes. Quartz crystal microbalance (QCM) measurement technique is one of the attractive techniques which has been used for monitoring the thin-film formation process. A sensitive QCM system was designed as to detect 0.1 Hz difference in resonance frequency and 10-7 change in energy dissipation values, which are the measures of the deposited mass and the film rigidity, respectively. PNIPAm microgels with the diameter around few hundred nanometers in water were produced via precipitation polymerization process. 5 MHz quartz crystals with functionalized gold surfaces were used for the deposition of the carrageenan molecules and microgels in the solutions which were slowly pumped through a flow cell. Interactions between charged carrageenan and microgel particles were monitored during the formation of the film layers, and the Sauerbrey masses of the deposited films were calculated. The critical phase transition temperatures around the LCST were detected during the heating and cooling cycles. It was shown that it is possible to monitor the interactions between PNIPAm microgels and biopolymer molecules, and it is also possible to specify the critical phase transition temperatures by using a QCM system.

Keywords: carrageenan, phase transitions, PNIPAm microgels, quartz crystal microbalance (QCM)

Procedia PDF Downloads 206
596 Molecular Dynamics Simulation Studies of Thermal Effects Created by High-Intensity, Ultra-Short Pulses Induced Cell Membrane Electroporation

Authors: Jiahui Song

Abstract:

The use of electric fields with high intensity (~ 100kV/cm or higher) and ultra short pulse durations (nanosecond range) has been a recent development. Most of the studies of electroporation have ignored possible thermal effects because of the small duration of the applied voltage pulses. However, it has been predicted membrane temperature gradients ranging from 0.2×109 to 109 K/m. This research focuses on thermal effects that drive for electroporative enhancements, even though the actual temperature values might not have changed appreciably from their equilibrium levels. The dynamics of pore formation with the application of an externally applied electric field is studied on the basis of molecular dynamics (MD) simulations using the GROMACS package. MD simulations of a lipid layer with constant electric field strength of 0.5 V/nm at 25 °C and 47 °C are implemented to simulate the appropriate thermal effects. The GROMACS provides the force fields for the lipid membranes, which is taken to comprise of dipalmitoyl-phosphatidyl-choline (DPPC) molecules. The water model mimicks the aqueous environment surrounding the membrane. Velocities of water and membrane molecules are generated randomly at each simulation run according to a Maxwellian distribution. The high background electric field is typically used in MD simulations to probe electroporation. It serves as an accelerated test of the pore formation process since low electric fields would take inordinately long simulation time. MD simulation shows no pore is formed in a 1-ns snapshot for a DPPC membrane set at a temperature of 25°C after a 0.5 V/nm electric field is applied. A nano-sized pore is clearly seen in a 0.75-ns snapshot on the same geometry, but with the membrane surfaces kept at temperatures of 47°C. And the pore increases at 1 ns. The MD simulation results suggest the possibility that the increase in temperature can result in different degrees of electrically stimulated bio-effects. The results points to the role of thermal effects in facilitating and accelerating the electroporation process.

Keywords: high-intensity, ultra-short, electroporation, thermal effects, molecular dynamics

Procedia PDF Downloads 27
595 Differences in Cognitive Functioning over the Course of Chemotherapy in Patients Suffering from Multiple Myeloma and the Possibility to Predict Their Cognitive State on the Basis of Biological Factors

Authors: Magdalena Bury-Kaminska, Aneta Szudy-Szczyrek, Aleksandra Nowaczynska, Olga Jankowska-Lecka, Marek Hus, Klaudia Kot

Abstract:

Introduction: The aim of the research was to determine the changes in cognitive functioning in patients with plasma cell myeloma by comparing patients’ state before the treatment and during chemotherapy as well as to determine the biological factors that can be used to predict patients’ cognitive state. Methods: The patients underwent the research procedure twice: before chemotherapy and after 4-6 treatment cycles. A psychological test and measurement of the following biological variables were carried out: TNF-α (tumor necrosis factor), IL-6 (interleukin 6), IL-10 (interleukin 10), BDNF (brain-derived neurotrophic factor). The following research methods were implemented: the Montreal Cognitive Assessment (MoCA), Battery of Tests for Assessing Cognitive Functions PU1, experimental and clinical trials based on the Choynowski’s Memory Scale, Stroop Color-Word Interference Test (SCWT), depression measurement questionnaire. Results: The analysis of the research showed better cognitive functions of patients during chemotherapy in comparison to the phase before it. Moreover, neurotrophin BDNF allows to predict the level of selected cognitive functions (semantic fluency and execution control) already at the diagnosis stage. After 4-6 cycles, it is also possible to draw conclusions concerning the extent of working memory based on the level of BDNF. Cytokine TNF-α allows us to predict the level of letter fluency during anti-cancer treatment. Conclusions: It is possible to presume that BDNF has a protective influence on patients’ cognitive functions and working memory and that cytokine TNF-α co-occurs with a diminished execution control and better material grouping in terms of phonological fluency. Acknowledgment: This work was funded by the National Science Center in Poland [grant no. 2017/27/N/HS6/02057.

Keywords: chemobrain, cognitive impairment, non−central nervous system cancers, hematologic diseases

Procedia PDF Downloads 130
594 Attenuation of Amyloid beta (Aβ) (1-42)-Induced Neurotoxicity by Luteolin

Authors: Dona Pamoda W. Jayatunga, Veer Bala Gupta, Eugene Hone, Ralph N. Martins

Abstract:

Being a neurodegenerative disorder, Alzheimer’s disease (AD) affects a majority of the elderly demented worldwide. The key risk factors for AD are age, metabolic syndrome, allele status of APOE gene, head injuries and lifestyle. The progressive nature of AD is characterized by symptoms of multiple cognitive deficits exacerbated over time, leading to death within a decade from clinical diagnosis. However, it is revealed that AD originates via a prodromal phase that spans from one to few decades before symptoms first manifest. The key pathological hallmarks of AD brains are deposition of amyloid beta (Aβ) plaques and neurofibrillary tangles (NFT). However, the yet unknown etiology of the disease fails to distinguish mitochondrial dysfunction between a cause or an outcome. The absence of early diagnosis tools and definite therapies for AD have permitted recruits of nutraceutical-based approaches aimed at reducing the risk of AD by modulating lifestyle or be used as preventive tools during AD prodromal state before widespread neurodegeneration begins. The objective of the present study was to investigate beneficial effects of luteolin, a plant-based flavone compound, against AD. The neuroprotective effects of luteolin on amyloid beta (Aβ) (1-42)-induced neurotoxicity was measured using cultured human neuroblastoma BE(2)-M17 cells. After exposure to 20μM Aβ (1-42) for 48 h, the neuroblastoma cells exhibited marked apoptotic death. Co-treatment of 20μM Aβ (1-42) with luteolin (0.5-5μM) significantly protected the cells against Aβ (1-42)-induced toxicity, as assessed by the MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2(4sulfophenyl)-2H-tetrazolium, inner salt; MTS] reduction assay and the lactate dehydrogenase (LDH) cell death assay. The results suggest that luteolin prevents Aβ (1-42)-induced apoptotic neuronal death. However, further studies are underway to determine its protective mechanisms in AD including the activity against tau hyperphosphorylation and mitochondrial dysfunction.

Keywords: Aβ (1-42)-induced toxicity, Alzheimer’s disease, luteolin, neuroblastoma cells

Procedia PDF Downloads 127
593 Comparative Numerical Simulations of Reaction-Coupled Annular and Free-Bubbling Fluidized Beds Performance

Authors: Adefarati Oloruntoba, Yongmin Zhang, Hongliang Xiao

Abstract:

An annular fluidized bed (AFB) is gaining extensive application in the process industry due to its efficient gas-solids contacting. But a direct evaluation of its reaction performance is still lacking. In this paper, comparative 3D Euler–Lagrange multiphase-particle-in-cell (MP-PIC) computations are performed to assess the reaction performance of AFB relative to a bubbling fluidized bed (BFB) in an FCC regeneration process. By using the energy-minimization multi-scale (EMMS) drag model with a suitable heterogeneity index, the MP-PIC simulation predicts the typical fountain region in AFB and solids holdup of BFB, which is consistent with an experiment. Coke combustion rate, flue gas and temperature profile are utilized as the performance indicators, while related bed hydrodynamics are explored to account for the different performance under varying superficial gas velocities (0.5 m/s, 0.6 m/s, and 0.7 m/s). Simulation results indicate that the burning rates of coke and its species are relatively the same in both beds, albeit marginal increase in BFB. Similarly, the shape and evolution time of flue gas (CO, CO₂, H₂O and O₂) curves are indistinguishable but match the coke combustion rates. However, AFB has high proclivity to high temperature-gradient as higher gas and solids temperatures are predicted in the freeboard. Moreover, for both beds, the effect of superficial gas velocity is only conspicuous on the temperature but negligible on combustion efficiency and effluent gas emissions due to constant gas volumetric flow rate and bed loading criteria. Cross-flow of solids from the annulus to the spout region as well as the high primary gas in the AFB directly assume the underlying mechanisms for its unique gas-solids hydrodynamics (pressure, solids holdup, velocity, mass flux) and local spatial homogeneity, which in turn influence the reactor performance. Overall, the study portrays AFB as a cheap alternative reactor to BFB for catalyst regeneration.

Keywords: annular fluidized bed, bubbling fluidized bed, coke combustion, flue gas, fountaining, CFD, MP-PIC, hydrodynamics, FCC regeneration

Procedia PDF Downloads 137
592 The Use of Bleomycin and Analogues to Probe the Chromatin Structure of Human Genes

Authors: Vincent Murray

Abstract:

The chromatin structure at the transcription start sites (TSSs) of genes is very important in the control of gene expression. In order for gene expression to occur, the chromatin structure at the TSS has to be altered so that the transcriptional machinery can be assembled and RNA transcripts can be produced. In particular, the nucleosome structure and positioning around the TSS has to be changed. Bleomycin is utilized as an anti-tumor agent to treat Hodgkin's lymphoma, squamous cell carcinoma, and testicular cancer. Bleomycin produces DNA damage in human cells and DNA strand breaks, especially double-strand breaks, are thought to be responsible for the cancer chemotherapeutic activity of bleomycin. Bleomycin is a large glycopeptide with molecular weight of approximately 1500 Daltons and hence its DNA strand cleavage activity can be utilized as a probe of chromatin structure. In this project, Illumina next-generation DNA sequencing technology was used to determine the position of DNA double-strand breaks at the TSSs of genes in intact cells. In this genome-wide study, it was found that bleomycin cleavage preferentially occurred at the TSSs of actively transcribed human genes in comparison with non-transcribed genes. There was a correlation between the level of enhanced bleomycin cleavage at TSSs and the degree of transcriptional activity. In addition, bleomycin was able to determine the position of nucleosomes at the TSSs of human genes. Bleomycin analogues were also utilized as probes of chromatin structure at the TSSs of human genes. In a similar manner to bleomycin, the bleomycin analogues 6′-deoxy-BLM Z and zorbamycin preferentially cleaved at the TSSs of human genes. Interestingly this degree of enhanced TSS cleavage inversely correlated with the cytotoxicity (IC50 values) of BLM analogues. This indicated that the degree of cleavage by bleomycin analogues at the TSSs of human genes was very important in the cytotoxicity of bleomycin and analogues. It also provided a deeper insight into the mechanism of action of this cancer chemotherapeutic agent since actively transcribed genes were preferentially targeted.

Keywords: anti-cancer activity, chromatin structure, cytotoxicity, gene expression, next-generation DNA sequencing

Procedia PDF Downloads 95
591 Defining New Limits in Hybrid Perovskites: Single-Crystal Solar Cells with Exceptional Electron Diffusion Length Reaching Half Millimeters

Authors: Bekir Turedi

Abstract:

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

Procedia PDF Downloads 26
590 Flow Reproduction Using Vortex Particle Methods for Wake Buffeting Analysis of Bluff Structures

Authors: Samir Chawdhury, Guido Morgenthal

Abstract:

The paper presents a novel extension of Vortex Particle Methods (VPM) where the study aims to reproduce a template simulation of complex flow field that is generated from impulsively started flow past an upstream bluff body at certain Reynolds number Re-Vibration of a structural system under upstream wake flow is often considered its governing design criteria. Therefore, the attention is given in this study especially for the reproduction of wake flow simulation. The basic methodology for the implementation of the flow reproduction requires the downstream velocity sampling from the template flow simulation; therefore, at particular distances from the upstream section the instantaneous velocity components are sampled using a series of square sampling-cells arranged vertically where each of the cell contains four velocity sampling points at its corner. Since the grid free Lagrangian VPM algorithm discretises vorticity on particle elements, the method requires transformation of the velocity components into vortex circulation, and finally the simulation of the reproduction of the template flow field by seeding these vortex circulations or particles into a free stream flow. It is noteworthy that the vortex particles have to be released into the free stream exactly at same rate of velocity sampling. Studies have been done, specifically, in terms of different sampling rates and velocity sampling positions to find their effects on flow reproduction quality. The quality assessments are mainly done, using a downstream flow monitoring profile, by comparing the characteristic wind flow profiles using several statistical turbulence measures. Additionally, the comparisons are performed using velocity time histories, snapshots of the flow fields, and the vibration of a downstream bluff section by performing wake buffeting analyses of the section under the original and reproduced wake flows. Convergence study is performed for the validation of the method. The study also describes the possibilities how to achieve flow reproductions with less computational effort.

Keywords: vortex particle method, wake flow, flow reproduction, wake buffeting analysis

Procedia PDF Downloads 285
589 Antioxidant Potential of Pomegranate Rind Extract Attenuates Pain, Inflammation and Bone Damage in Experimental Rats

Authors: Ritu Karwasra, Surender Singh

Abstract:

Inflammation is an important physiological response of the body’s self-defense system that helps in eliminating and protecting organism from harmful stimuli and in tissue repair. It is a highly regulated protective response which helps in eliminating the initial cause of cell injury, and initiates the process of repair. The present study was designed to evaluate the ameliorative effect of pomegranate rind extract on pain and inflammation. Hydroalcoholic standardized rind extract of pomegranate at doses 50, 100 and 200 mg/kg and indomethacin (3 mg/kg) was tested against eddy’s hot plate induced thermal algesia, carrageenan (acute inflammation) and Complete Freund’s Adjuvant (chronic inflammation) induced models in Wistar rats. Parameters analyzed were inhibition of paw edema, measurement of joint diameter, levels of GSH, TBARS, SOD, TNF-α, radiographic imaging, tissue histology and synovial expression of pro-inflammatory cytokine receptor (TNF-R1). Radiological and light microscopical analysis were carried out to find out the bone damage in CFA-induced chronic inflammatory model. Findings of the present study revealed that pomegranate rind extract at a dose of 200 mg/kg caused a significant (p<0.05) reduction in paw swelling in both the inflammatory models. Nociceptive threshold was also significantly (p<0.05) improved. Immunohistochemical analysis of TNF-R1 in CFA-induced group showed elevated level, whereas reduction in level of TNF-R1 was observed in pomegranate (200 mg/kg). Henceforth, we might say that pomegranate produced a dose-dependent reduction in inflammation and pain along with the reduction in levels of oxidative stress markers and tissue histology, and the effect was found to be comparable to that of indomethacin. Thus, it can be concluded that pomegranate is a potential therapeutic target in the pathogenesis of inflammation and pain, and punicalagin is the major constituents found in rind extract might be responsible for the activity.

Keywords: carrageenan, inflammation, nociceptive-threshold, pomegranate, histopathology

Procedia PDF Downloads 190
588 In vivo Activity of Pathogenic Bacteria on Natural Polyphenolic Compounds

Authors: Lubna Azmi, Ila Shukla, Shyam Sundar Gupta, Padam Kant, Ch. V. Rao

Abstract:

Gastric ulcer is a major global health threat, and it is the leading cause of stomach cancer death worldwide. Helicobacter pylori bacteriumis the most important etiologic factor for gastric ulcer. This infection is highly pervasive in South Asian developing countries, especially in India, Nepal, Srilanka etc. due to diversification in geographic area. Pathophysiology of gastric mucosal damage associated with non-invasive bacterium has not justified in detail, but it leads to change in histopathology, immunochemistry of the gastric and duodenal reason of host. The mechanism responsible for bacteria tissue tropism and mucosal damage in stomach variance during the disease is not clearly described and understood scientifically in treatment and control of pathogenic organisms. Polyphenols are secondary metabolites of plants and are generally involved in defense against aggression by pathogens. 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxychromen-4-one and 1-hydroxy-5,7-dimethoxy-2-naphthalene-carboxaldehyde are polyphenolic compound obtained from popular Indian medicinal plants ghavpatta (ArgeriaspeciosaLinn.f) andBael (Aeglemarmelos) have long been used in traditional Ayurvedic Indian medicine for various diseases. They have promising effects on ulcer, as detailed investigation has made in our laboratory. Therefore, the aim of present study is to explore membrane –dependent morphogenesis of H. pylori and associated apoptosis-mediated cell death. Based on this we analyzed immune gene expression in stomach of experimental animals with H. pylori, using quantitative reverse transcription polymerase chain reaction(q RT-PCR). This revealed rapid induction of prostaglandin, interferon I (INF-I), interferon II (INF-II) and INF-I associated genes in the infected animal. Ultrastructural changes associated with H. pylori will be taken for advanced studies. This investigation shows that the biomarkers eradicate H. pylori bacterium caused gastric ulcer which is a major risk factor for gastric cancer.

Keywords: gastric ulcer, Helicobacter pylori, immunochemistry, polyphenols

Procedia PDF Downloads 349
587 Insight into the Electrocatalytic Activities of Nitrogen-Doped Graphyne and Graphdiyne Families: A First-Principles Study

Authors: Bikram K. Das, Kalyan K. Chattopadhyay

Abstract:

The advent of 2-D materials in the last decade has induced a fresh spur of growth in fuel cell technology as these materials have some highly promising traits that can be exploited to felicitate Oxygen Reduction Reaction (ORR) in an efficient way. Among the various 2-D carbon materials, graphyne (Gy) and graphdiyne (Gdy)1 with their intrinsic non-uniform charge distribution holds promises in this purpose and it is expected2 that substitutional Nitrogen (N) doping could further enhance their efficiency. In this regard, dispersive force corrected density functional theory is used to map the oxygen reduction reaction (ORR) kinetics of five different kinds of N doped graphyne and graphdiyne systems (namely αGy, βGy, γGy, RGy and 6,6,12Gy and Gdy) in alkaline medium. The best doping site for each of the Gy/ Gdy system is determined comparing the formation energies of the possible doping configurations. Similarly, the best di-oxygen (O₂) adsorption sites for the doped systems are identified by comparing the adsorption energies. O₂ adsorption on all N doped Gy/ Gdy systems is found to be energetically favorable. ORR on a catalyst surface may occur either via the Eley-Rideal (ER) or the Langmuir–Hinschelwood (LH) pathway. Systematic studies performed on the considered systems reveal that all of them favor the ER pathway. Further, depending on the nature of di-oxygen adsorption ORR can follow either associative or dissociative mechanism; the possibility of occurrence of both the mechanisms is tested thoroughly for each N doped Gy/ Gdy. For the ORR process, all the Gy/Gdy systems are observed to prefer the efficient four-electron pathway but the expected monotonically exothermic reaction pathway is found only for N doped 6,6,12Gy and RGy following the associative pathway and for N doped βGy, γGy and Gdy following the dissociative pathway. Further computation performed for these systems reveals that for N doped 6,6,12Gy, RGy, βGy, γGy and Gdy the overpotentials are 1.08 V, 0.94 V, 1.17 V, 1.21 V and 1.04 V respectively depicting N doped RGy is the most promising material, to carry out ORR in alkaline medium, among the considered ones. The stability of the ORR intermediate states with the variation of pH and electrode potentials is further explored with Pourbiax diagrams and the activities of these systems in the alkaline medium are compared with the prior reported B/N doped identical systems for ORR in an acidic medium in terms of a common descriptor.

Keywords: graphdiyne, graphyne, nitrogen-doped, ORR

Procedia PDF Downloads 98
586 Comparative Evaluation of Seropositivity and Patterns Distribution Rates of the Anti-Nuclear Antibodies in the Diagnosis of Four Different Autoimmune Collagen Tissue Diseases

Authors: Recep Kesli, Onur Turkyilmaz, Cengiz Demir

Abstract:

Objective: Autoimmune collagen diseases occur with the immune reactions against the body’s own cell or tissues which cause inflammation and damage the tissues and organs. In this study, it was aimed to compare seropositivity rates and patterns of the anti-nuclear antibodies (ANA) in the diagnosis of four different autoimmune collagen tissue diseases (Rheumatoid Arthritis-RA, Systemic Lupus Erythematous-SLE, Scleroderma-SSc and Sjogren Syndrome-SS) with each other. Methods: One hundred eighty-eight patients applied to different clinics in Afyon Kocatepe University ANS Practice and Research Hospital between 11.07.2014 and 14.07.2015 that thought the different collagen disease such as RA, SLE, SSc and SS have participated in the study retrospectively. All the data obtained from the patients participated in the study were evaluated according to the included criteria. The historical archives belonging to the patients have been screened, assessed in terms of ANA positivity. The obtained data was analysed by using the descriptive statistics; chi-squared, Fischer's exact test. The evaluations were performed by SPSS 20.0 version and p < 0.05 level was considered as significant. Results: Distribution rates of the totally one hundred eighty-eight patients according to the diagnosis were found as follows: 82 (43.6%) were RA, 38 (20.2%) were SLE, 22 (11.7%) were SSc, and 46 (24.5%) were SS. Distribution of ANA positivity rates according to the collagen tissue diseases were found as follows; for RA were 54 (65,9 %), for SLE were 36 (94,7 %), for SSc were 18 (81,8 %), and for SS were 43 (93,5 %). Rheumatoid arthritis should be evaluated and classified as a different class among all the other investigated three autoimmune illnesses. ANA positivity rates were found as differently higher (91.5 %) in the SLE, SSc, and SS, from the RA (65.9 %). Differences at ANA positivity rates for RA and the other three diseases were found as statistically significant (p=0.015). Conclusions: Systemic autoimmune illnesses show broad spectrum. ANA positivity was found as an important predictor marker in the diagnosis of the rheumatologic illnesses. ANA positivity should be evaluated as more valuable and sensitive a predictor diagnostic marker in the laboratory findings of the SLE, SSc, and SS according to RA.

Keywords: antinuclear antibody (ANA), rheumatoid arthritis, scleroderma, Sjogren syndrome, systemic lupus Erythemotosus

Procedia PDF Downloads 206
585 Pale, Soft, Exudative (PSE) Turkey Meat in a Brazilian Commercial Processing Plant

Authors: Danielle C. B. Honorato, Rafael H. Carvalho, Adriana L. Soares, Ana Paula F. R. L. Bracarense, Paulo D. Guarnieri, Massami Shimokomaki, Elza I. Ida

Abstract:

Over the past decade, the Brazilian production of turkey meat increased by more than 50%, indicating that the turkey meat is considered a great potential for the Brazilian economy contributing to the growth of agribusiness at the marketing international scenario. However, significant color changes may occur during its processing leading to the pale, soft and exudative (PSE) appearance on the surface of breast meat due to the low water holding capacity (WHC). Changes in PSE meat functional properties occur due to the myofibrils proteins denaturation caused by a rapid postmortem glycolysis resulting in a rapid pH decline while the carcass temperature is still warm. The aim of this study was to analyze the physical, chemical and histological characteristics of PSE turkey meat obtained from a Brazilian commercial processing plant. The turkey breasts samples were collected (n=64) at the processing line and classified as PSE at L* ≥ 53 value. The pH was also analyzed after L* measurement. In sequence, PSE meat samples were evaluated for WHC, cooking loss (CL), shear force (SF), myofibril fragmentation index (MFI), protein denaturation (PD) and histological evaluation. The abnormal color samples presented lower pH values, 16% lower fiber diameter, 11% lower SF and 2% lower WHC than those classified as normal. The CL, PD and MFI were, respectively, 9%, 18% and 4% higher in PSE samples. The Pearson correlation between the L* values and CL, PD and MFI was positive, while that SF and pH values presented negative correlation. Under light microscopy, a shrinking of PSE muscle cell diameter was approximately 16% shorter in relation to normal samples and an extracellular enlargement of endomysium and perimysium sheaths as the consequence of higher water contents lost as observed previously by lower WHC values. Thus, the results showed that PSE turkey breast meat presented significant changes in their physical, chemical and histological characteristics that may impair its functional properties.

Keywords: functional properties, histological evaluation, meat quality, PSE

Procedia PDF Downloads 431
584 Electrical and Structural Properties of Solid Electrolyte Systems

Authors: Yasin Polat, Yılmaz Dağdemir, Mehmet Arı

Abstract:

Samarium (III) oxide and Ytterbium (III) oxide doped Bismuth trioxide solid solutions, the nano ceramic (Bi2O3)1-x-y(Sm2O3)x(Yb2O3)y ternary system were obtained with x=5, 20 mol %, and y=5, 20 mol % dopant concentrations have been synthesized in air atmosphere with solid state reaction. Temperature dependent electrical conductivity of the samples have been investigated by 4-point probe technique by heating and cooling process. Doped-Bi2O3 materials of solid electrolyte systems are good oxygen anions O2-conductors which have collected much attention as potential solid ceramic electrolytes for solid oxide fuel cells (SOFCs) because of their relatively high oxygen ionic conductivity at lower temperatures.(Bi2O3)-based electrolytes have also wide other technological applications in devices with high economical interest such as oxygen sensors, ceramic membranes for oxygen separation, oxygen pumps, catalyzing of some heterogeneous reactions, partial oxidation of the hydrocarbons, and additive material in paints. In recent years, many experimental researches have mostly focused on improving of the Bi-based electrolytes which have high oxide ionic conductivity at low temperatures and better performance as alternatives to traditional stabilized zirconia has taken place. Generally, these systems are much better solid electrolytes than well-known stabilized zirconia, because some of the bismuth trioxide phases exhibit higher ion conductivity than other oxide ionic conductors. Crystal structure of the Nano ceramic (Bi2O3)1-x-y(Sm2O3)x(Yb2O3)y has been determined by X-Ray powder diffractions (XRD) measurements before and after electrical conductivity measurements of the samples. Surface and grain structure properties of the samples were determined by SEM analysis. The samples which synthesized in this study can be used in industrial applications such as electrolytes of the solid oxide fuel cells (SOFC).

Keywords: 4-point probe technique, bismuth trioxide, solid state reaction, solid oxide fuel cell

Procedia PDF Downloads 262
583 Highly Active, Non-Platinum Metal Catalyst Material as Bi-Functional Air Cathode in Zinc Air Battery

Authors: Thirupathi Thippani, Kothandaraman Ramanujam

Abstract:

Current research on energy storage has been paid to metal-air batteries, because of attractive alternate energy source for the future. Metal – air batteries have the probability to significantly increase the power density, decrease the cost of energy storage and also used for a long time due to its high energy density, low-level pollution, light weight. The performance of these batteries mostly restricted by the slow kinetics of the oxygen reduction reaction (ORR) and oxygen evolution reaction (OER) on cathode during battery discharge and charge. The ORR and OER are conventionally carried out with precious metals (such as Pt) and metal oxides (such as RuO₂ and IrO₂) as catalysts separately. However, these metal-based catalysts are regularly undergoing some difficulties, including high cost, low selectivity, poor stability and unfavorable to environmental effects. So, in order to develop the active, stable, corrosion resistance and inexpensive bi-functional catalyst material is mandatory for the commercialization of zinc-air rechargeable battery technology. We have attempted and synthesized non-precious metal (NPM) catalysts comprising cobalt and N-doped multiwalled carbon nanotubes (N-MWCNTs-Co) were synthesized by the solid-state pyrolysis (SSP) of melamine with Co₃O₄. N-MWCNTs-Co acts as an excellent electrocatalyst for both the oxygen reduction reaction (ORR) and the oxygen evolution reaction (OER), and hence can be used in secondary metal-air batteries and in unitized regenerative fuel cells. It is important to study the OER and ORR at high concentrations of KOH as most of the metal-air batteries employ KOH concentrations > 4M. In the first 16 cycles of the zinc-air battery while using N-MWCNTs-Co, 20 wt.% Pt/C or 20 wt.% IrO₂/C as air electrodes. In the ORR regime (the discharge profile of the zinc-air battery), the cell voltage exhibited by N-MWCNTs-Co was 44 and 83 mV higher (based on 5th cycle) in comparison to of 20 wt.% Pt/C and 20 wt.% IrO₂/C respectively. To demonstrate this promise, a zinc-air battery was assembled and tested at a current density of 0.5 Ag⁻¹ for charge-discharge 100 cycles.

Keywords: oxygen reduction reaction (ORR), oxygen evolution reaction(OER), non-platinum, zinc air battery

Procedia PDF Downloads 206
582 The Efficacy of Pre-Hospital Packed Red Blood Cells in the Treatment of Severe Trauma: A Retrospective, Matched, Cohort Study

Authors: Ryan Adams

Abstract:

Introduction: Major trauma is the leading cause of death in 15-45 year olds and a significant human, social and economic costs. Resuscitation is a stalwart of trauma management, especially in the pre-hospital environment and packed red blood cells (pRBC) are being increasingly used with the advent of permissive hypotension. The evidence in this area is lacking and further research is required to determine its efficacy. Aim: The aim of this retrospective, matched cohort study was to determine if major trauma patients, who received pre-hospital pRBC, have a difference in their initial emergency department cardiovascular status; when compared with injury-profile matched controls. Methods: The trauma databases of the Royal Brisbane and Women's Hospital, Royal Children's Hospital (Herston) and Queensland Ambulance Service were accessed and major trauma patient (ISS>12) data, who received pre-hospital pRBC, from January 2011 to August 2014 was collected. Patients were then matched against control patients that had not received pRBC, by their injury profile. The primary outcomes was cardiovascular status; defined as shock index and Revised Trauma Score. Results: Data for 25 patients who received pre-hospital pRBC was accessed and the injury profiles matched against suitable controls. On admittance to the emergency department, a statistically significant difference was seen in the blood group (Blood = 1.42 and Control = 0.97, p-value = 0.0449). However, the same was not seen with the RTS (Blood = 4.15 and Control 5.56, p-value = 0.291). Discussion: A worsening shock index and revised trauma score was associated with pre-hospital administration of pRBC. However, due to the small sample size, limited matching protocol and associated confounding factors it is difficult to draw any solid conclusions. Further studies, with larger patient numbers, are required to enable adequate conclusions to be drawn on the efficacy of pre-hospital packed red blood cell transfusion.

Keywords: pre-hospital, packed red blood cells, severe trauma, emergency medicine

Procedia PDF Downloads 370
581 Cost Effective Microfabrication Technique for Lab on Chip (LOC) Devices Using Epoxy Polymers

Authors: Charmi Chande, Ravindra Phadke

Abstract:

Microfluidics devices are fabricated by using multiple fabrication methods. Photolithography is one of the common methods wherein SU8 is widely used for making master which in turn is used for making working chip by the process of soft lithography. The high-aspect ratio features of SU-8 makes it suitable to be used as micro moulds for injection moulding, hot embossing, and moulds to form polydimethylsiloxane (PDMS) structures for bioMEMS (Microelectromechanical systems) applications. But due to high cost, difficulty in procuring and need for clean room, restricts the use of this polymer especially in developing countries and small research labs. ‘Bisphenol –A’ based polymers in mixture with curing agent are used in various industries like Paints and coatings, Adhesives, Electrical systems and electronics, Industrial tooling and composites. We present the novel use of ‘Bisphenol – A’ based polymer in fabricating micro channels for Lab On Chip(LOC) devices. The present paper describes the prototype for production of microfluidics chips using range of ‘Bisphenol-A’ based polymers viz. GY 250, ATUL B11, DER 331, DER 330 in mixture with cationic photo initiators. All the steps of chip production were carried out using an inexpensive approach that uses low cost chemicals and equipment. This even excludes the need of clean room. The produced chips using all above mentioned polymers were validated with respect to height and the chip giving least height was selected for further experimentation. The lowest height achieved was 7 micrometers by GY250. The cost of the master fabricated was $ 0.20 and working chip was $. 0.22. The best working chip was used for morphological identification and profiling of microorganisms from environmental samples like soil, marine water and salt water pan sites. The current chip can be adapted for various microbiological screening experiments like biochemical based microbial identification, studying uncultivable microorganisms at single cell/community level.

Keywords: bisphenol–A based epoxy, cationic photoinitiators, microfabrication, photolithography

Procedia PDF Downloads 262
580 Construction of Genetic Recombinant Yeasts with High Environmental Tolerance by Accumulation of Trehalose and Detoxication of Aldehyde

Authors: Yun-Chin Chung, Nileema Divate, Gen-Hung Chen, Pei-Ru Huang, Rupesh Divate

Abstract:

Many environmental factors, such as glucose concentration, ethanol, temperature, osmotic pressure and pH, decrease the production rate of ethanol using yeast as a starter. Fermentation starters with high tolerance to various stresses are always demanded for brewing industry. Trehalose, a storage carbohydrate in cell wall of yeast, plays an important role in tolerance of environmental stress by preserving integrity of plasma membrane and stabilizing proteins. Furan aldehydes are toxic to yeast and the growth rate of yeast is significantly reduced if furan aldehydes were present in the fermentation medium. In yeast, aldehyde reductase is involved in the detoxification of reactive aldehydes and consequently the growth of yeast is improved. The aims of this study were to construct a genetic recombinant Saccharomyces cerevisiae or Pichia pastoris with furfural and HMF degrading and high ethanol tolerance capacities. Yeast strains were engineered by genetic recombination for overexpression of trehalose-6-phosphate synthase gene (tps1) and aldehyde reductase gene (ari1). TPS1 gene was cloned from S. cerevisiae by reverse transcription-polymerase chain reaction (RT-PCR) and then ligated with pGAPZαC vector. The constructed vector, pGAPZC-tps1, was transformed to recombinant yeasts strain with overexpression of ari1. The transformants with pGAPZC-tps1-ari1 were generated called STA (S. cerevisiae) and PTA (P. pastoris) with overexpression of tps1, ari1. PCR with tps1-specific primers and western blot with his-tag confirmed the gene insertion and protein expression of tps1 in the transformants, respectively. The neutral trehalase gene (nth1) of STA was successfully deleted and the novel strain STAΔN will be used for further study, including the measurement of trehalose concentration and ethanol, furfural tolerance assay.

Keywords: genetic recombinant, yeast, ethanol tolerance, trehalase, aldehyde reductase

Procedia PDF Downloads 398
579 Biosurfactants Production by Bacillus Strain from an Environmental Sample in Egypt

Authors: Mervat Kassem, Nourhan Fanaki, F. Dabbous, Hamida Abou-Shleib, Y. R. Abdel-Fattah

Abstract:

With increasing environmental awareness and emphasis on a sustainable society in harmony with the global environment, biosurfactants are gaining prominence and have already taken over for a number of important industrial uses. They are produced by living organisms, for examples Pseudomonas aeruginosa which produces rhamnolipids, Candida (formerly Torulopsis) bombicola, which produces high yields of sophorolipids from vegetable oils and sugars and Bacillus subtilis which produces a lipopeptide called surfactin. The main goal of this work was to optimize biosurfactants production by an environmental Gram positive isolate for large scale production with maximum yield and low cost. After molecular characterization, phylogenetic tree was constructed where it was found to be B. subtilis, which close matches to B. subtilis subsp. subtilis strain CICC 10260. For optimizing its biosurfactants production, sequential statistical design using Plackett-Burman and response surface methodology, was applied where 11 variables were screened. When analyzing the regression coefficients for the 11 variables, pH, glucose, glycerol, yeast extract, ammonium chloride and ammonium nitrate were found to have a positive effect on the biosurfactants production. Ammonium nitrate, pH and glucose were further studied as significant independent variables for Box-Behnken design and their optimal levels were estimated and were found to be 7.328 pH value, 3 g% glucose and 0.21g % ammonium nitrate yielding high biosurfactants concentration that reduced the surface tension of the culture medium from 72 to 18.16 mN/m. Next, kinetics of cell growth and biosurfactants production by the tested B. subtilis isolate, in bioreactor was compared with that of shake flask where the maximum growth and specific growth (µ) in the bioreactor was higher by about 25 and 53%, respectively, than in shake flask experiment, while the biosurfactants production kinetics was almost the same in both shake flask and bioreactor experiments.

Keywords: biosurfactants, B. subtilis, molecular identification, phylogenetic trees, Plackett-Burman design, Box-Behnken design, 16S rRNA

Procedia PDF Downloads 383
578 Self-Healing Hydrogel Triggered by Magnetic Microspheres to Control Glutathione Release for Cartilage Repair

Authors: I-Yun Cheng, Min-Yu Chiang, Shwu-Jen Chang, San-Yuan Chen

Abstract:

Osteoarthritis (OA) is among the most challenging joint diseases, and as far as we know, there is currently no exact and effective cure for it because it has low self-repair ability due to lack of blood vessels and low cell density in articular cartilage. So far, there have been several methods developed to treat cartilage disorder. The most common method is to treat the high molecular weight of hyaluronic acid (HA) injection, but it will degrade after a period of time, so the patients need to inject HA repeatedly. In recent years, self-healing hydrogel has drawn considerable attention because it can recover its initial mechanical properties after damaged and further increase the lifetime of the hydrogel. Here, we aim to develop a self-healable composite hydrogel combined with magnetic microspheres to trigger glutathione(GSH) release for promoting cartilage repair. We use HA-cyclodextrin (CD) as host polymer and poly(acrylic acid)-ferrocene (pAA-Fc) as guest polymer to form the self-healable HA-pAA hydrogel by host and guest interaction where various graft amount of pAA-Fc (pAA:Fc= 1:2, 1:1.5, 1:1, 2:1, 4:1) was conducted to develop different mechanical strength hydrogel. The rheology analysis showed that the 4:1 of pAA-Fc has higher mechanical strength than other formulations. On the other hand, iron oxide nanoparticle, poly(lactic-co-glycolic acid) (PLGA) and polyethyleneimine (PEI) were used to synthesize porous magnetic microspheres via double emulsification water-in-oil-in-water (W/O/W) to increase GSH loading which acted as a reductant to control the hydrogel crosslink density and promote hydrogel self-healing. The results show that the porous magnetic microspheres can be loaded with 70% of GSH and sustained release about 50% of GSH after 24 hours. More importantly, the HA-pAA composite hydrogel can self-heal rapidly within 24 hours when suffering external force destruction by releasing GSH from the magnetic microspheres. Therefore, the developed the HA-pAA composite hydrogel combined with GSH-loaded magnetic microspheres can be in-vivo guided to damaged OA surface for inducing the cartilage repair by controlling the crosslinking of self-healing hydrogel via GSH release.

Keywords: articular cartilage, magnetic microsphere, osteoarthritis, self-healing hydrogel

Procedia PDF Downloads 106
577 Transcriptome Analysis for Insights into Disease Progression in Dengue Patients

Authors: Abhaydeep Pandey, Shweta Shukla, Saptamita Goswami, Bhaswati Bandyopadhyay, Vishnampettai Ramachandran, Sudhanshu Vrati, Arup Banerjee

Abstract:

Dengue virus infection is now considered as one of the most important mosquito-borne infection in human. The virus is known to promote vascular permeability, cerebral edema leading to Dengue hemorrhagic fever (DHF) or Dengue shock syndrome (DSS). Dengue infection has known to be endemic in India for over two centuries as a benign and self-limited disease. In the last couple of years, the disease symptoms have changed, manifesting severe secondary complication. So far, Delhi has experienced 12 outbreaks of dengue virus infection since 1997 with the last reported in 2014-15. Without specific antivirals, the case management of high-risk dengue patients entirely relies on supportive care, involving constant monitoring and timely fluid support to prevent hypovolemic shock. Nonetheless, the diverse clinical spectrum of dengue disease, as well as its initial similarity to other viral febrile illnesses, presents a challenge in the early identification of this high-risk group. WHO recommends the use of warning signs to identify high-risk patients, but warning signs generally appear during, or just one day before the development of severe illness, thus, providing only a narrow window for clinical intervention. The ability to predict which patient may develop DHF and DSS may improve the triage and treatment. With the recent discovery of high throughput RNA sequencing allows us to understand the disease progression at the genomic level. Here, we will collate the results of RNA-Sequencing data obtained recently from PBMC of different categories of dengue patients from India and will discuss the possible role of deregulated genes and long non-coding RNAs NEAT1 for development of disease progression.

Keywords: long non-coding RNA (lncRNA), dengue, peripheral blood mononuclear cell (PBMC), nuclear enriched abundant transcript 1 (NEAT1), dengue hemorrhagic fever (DHF), dengue shock syndrome (DSS)

Procedia PDF Downloads 285
576 Stereological Evaluation of Liver of Rabbit Fetuses After Transplantation of Human Wharton’s Jelly-Derived Mesenchymal Stromal/Stem Cells

Authors: Zahra Khodabandeh, Leila Rezaeian, Mohammad Amin Edalatmanesh, Asghar Mogheiseh, Nader Tanideh, Mehdi Dianatpour, Shahrokh Zare, Hossein Bordbar, Neda Baghban, Amin Tamadon

Abstract:

Background: In-utero xenotransplantation of stem cells in abnormal fetuses effectively treats several genetic illnesses. Objective: The current research aimed to evaluate structural and morphological alterations in the liver of rabbit fetuses following xenotransplantation of human Wharton’s jelly-derived mesenchymal stromal cells (hWJ-MSCs) using a stereological technique. Methods: hWJ-MSCs were isolated from the human umbilical cord, and their authenticity was established by flow cytometry and differentiation. At gestational day 14, the rabbits were anesthetized, and hWJ-MSCs were injected into the uteri of 24 fetuses. Twenty-two fetuses were born successfully. Ten rabbit liver specimens were prepared from injected fetuses, including eight rabbits on day three following birth and two rabbits on the 21st post-natal day. The non-injected fetuses were considered positive controls. The livers of the control and hWJ-MSCs-treated rabbits were fixed, processed, stained, and examined through stereological approaches. Results: In the hWJ-MSCs-treated group, the mean liver weight and volume increased by 42% and 78% compared to the control group. The total volume of the hepatocytes increased by 63% and that of sinusoids by threefold in the treated rabbits. The total volume of the central veins increased by 70%. The total number corresponding to hepatocytes in the experimental group increased by 112% compared to the rabbits in the control. The total volume of the hepatocyte nuclei in the experimental group increased by 117% compared to the rabbits in the control. Conclusion: After xenotransplantation of human MSCs, host tissue microenvironments (here, the rabbit liver) were altered, and these included quantitative factors corresponding to the liver tissue and hepatocyte morphometric indices.

Keywords: xenotransplantation, mesenchymal stromal, stem cell, Wharton ‘s jelly, liver

Procedia PDF Downloads 57
575 Motion Planning and Simulation Design of a Redundant Robot for Sheet Metal Bending Processes

Authors: Chih-Jer Lin, Jian-Hong Hou

Abstract:

Industry 4.0 is a vision of integrated industry implemented by artificial intelligent computing, software, and Internet technologies. The main goal of industry 4.0 is to deal with the difficulty owing to competitive pressures in the marketplace. For today’s manufacturing factories, the type of production is changed from mass production (high quantity production with low product variety) to medium quantity-high variety production. To offer flexibility, better quality control, and improved productivity, robot manipulators are used to combine material processing, material handling, and part positioning systems into an integrated manufacturing system. To implement the automated system for sheet metal bending operations, motion planning of a 7-degrees of freedom (DOF) robot is studied in this paper. A virtual reality (VR) environment of a bending cell, which consists of the robot and a bending machine, is established using the virtual robot experimentation platform (V-REP) simulator. For sheet metal bending operations, the robot only needs six DOFs for the pick-and-place or tracking tasks. Therefore, this 7 DOF robot has more DOFs than the required to execute a specified task; it can be called a redundant robot. Therefore, this robot has kinematic redundancies to deal with the task-priority problems. For redundant robots, Pseudo-inverse of the Jacobian is the most popular motion planning method, but the pseudo-inverse methods usually lead to a kind of chaotic motion with unpredictable arm configurations as the Jacobian matrix lose ranks. To overcome the above problem, we proposed a method to formulate the motion planning problems as optimization problem. Moreover, a genetic algorithm (GA) based method is proposed to deal with motion planning of the redundant robot. Simulation results validate the proposed method feasible for motion planning of the redundant robot in an automated sheet-metal bending operations.

Keywords: redundant robot, motion planning, genetic algorithm, obstacle avoidance

Procedia PDF Downloads 124