Search results for: bloodstream expression site
3748 Targeting Matrix Metalloprotease-9 to Reduce Coronary Artery Manifestations of Kawasaki’s Disease
Authors: Mohammadjavad Sotoudeheian, Navid Farahmandian
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Kawasaki disease (KD) is the primary cause of acquired pediatric heart disease as an acute vasculitis. In children with prolonged fever, rash, and inflammation of the mucosa KD must be considered as a clinical diagnosis. There is a persuasive suggestion of immune-mediated damage as the pathophysiologic cascade of KD. For example, the invasion of cytotoxic T-cells supports a viral etiology and the inflammasome of the innate immune system is a critical component in the vasculitis formation in KD. Animal models of KD propose the cytokine profiles, such as increased IL-1 and GM-CSF, which cause vascular damage. CRP and IFN-γ elevated expression and the upregulation of IL-6, and IL-10 production are also described in previous studies. Untreated KD is a critical risk factor for coronary artery diseases and myocardial infarction. Vascular damage may encompass amplified T-cell activity. SMAD3 is an essential molecule in down-regulating T-cells and increasing expression of FoxP3. It has a critical effect in the differentiation of regulatory T-cells. The discrepancy of regulatory T-cells and pro-inflammatory Th17 has been studied in acute coronary syndrome during KD. However in the coronary artery damaged lymphocytes and IgA plasma cells are seen at the lesion locations, the major immune cells in the coronary lesions are monocytes/macrophages and neutrophils. These cells secrete TNF-α, and activates matrix metalloprotease (MMP)-9, reducing the integrity of vessels and prompting patients to arise aneurysm. MMPs can break down the components of the extracellular matrix and assist immune cell movement. IVIG as an effective form of treatment clarified the role of the immune system, which may target pathogenic antigens and regulate cytokine production. Several reports have revealed that in the coronary arteries, high expression of MMP-9 in monocyte/macrophage results in pathologic cascades. Curcumin is a potent antioxidant and anti-inflammatory molecule. Curcumin decreases the production of reactive oxygen and nitrogen species and inhibits transcription factors like AP-1 and NF-κB. Curcumin also contains the characteristics of inhibitory effects on MMPs, especially MMP-9. The upregulation of MMP-9 is an important cellular response. Curcumin treatment caused a reverse effect and down-regulates MMP-9 gene expression which may fund the anti-inflammatory effect. Curcumin inhibits MMP-9 expression via PKC and AMPK-dependent pathways in Human monocytes cells. Elevated expression and activity of MMP-9 are correlated with advanced vascular lesions. AMPK controls lipid metabolism and oxidation, and protein synthesis. AMPK is also necessary for the MMP-9 activity and THP-1 cell adhesion to endothelial cells. Curcumin was shown to inhibit the activation of AMPKα. Compound C (AMPK inhibitor) inhibits MMP-9 expression level. Therefore, through inactivating AMPKs and PKC, curcumin decreases the MMP-9 level, which results in inhibiting monocyte/macrophage differentiation. Compound C also suppress the phosphorylation of three major classes of MAP kinase signaling, suggesting that curcumin may suppress MMP-9 level by inactivation of MAPK pathways. MAPK cascades are activated to induce the expression of MMP-9. Curcumin inhibits MAPKs phosphorylation, which contributes to the down-regulation of MMP-9. This study demonstrated that the potential inhibitory properties of curcumin over MMP-9 lead to a therapeutic strategy to reduce the risk of coronary artery involvement during KD.Keywords: MMP-9, coronary artery aneurysm, Kawasaki’s disease, curcumin, AMPK, immune system, NF-κB, MAPK
Procedia PDF Downloads 3043747 Production of Camel Nanobodies against of Anti-Morphine-3-Glucuronide for the Development of a Biosensor for Detecting Illicit Drug
Authors: Shirin Jalili, Sadegh Hasannia, Hadi Shirzad, Afshin Khara
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Morphine is one of the most medicinally important analgesics and narcotics. Structurally, it is classified as an alkaloid because of the presence of nitrogen. Its structure is similar to that of codeine, thebaine, and heroin. An immunoassay to accurately discriminate between these analogous alkaloids would be highly beneficial. A key factor for such an assay is specificity with high sensitivity, which is totally dependent on the antibody employed. However, most antibodies against haptens are polyclonal serum antibodies that exhibit significant cross-reactivities with closely related compounds. The camel-derived single-chain antibody fragments (VHH) are the smallest molecules with antigen-binding capacity, possessing unique properties compared to other conventional antibodies. In this study, a library containing the VHH genes of a camel immunized with with morphine conjugated BSA following phage display technology was generated. By screening the camel-derived variable region of the heavy chain cDNA phage display library with the ability to bind the desired hapten, we obtained some nanobodies that recognize this hapten. Phage display expression of the Nbs from this library and pannings against this hapten resulted in a clear enrichment of four distinct Nb-displaying phages with specificity for morphine that could be a potential target site for the development of new strategies for the development of a biosensor for detecting illicit drug.Keywords: phage display, nanobody, Morphine-3, glucuronide, ELISA, biosensor
Procedia PDF Downloads 4253746 Evaluating Energy Transition of a complex of buildings in a historic site of Rome toward Zero-Emissions for a Sustainable Future
Authors: Silvia Di Turi, Nicolandrea Calabrese, Francesca Caffari, Giulia Centi, Francesca Margiotta, Giovanni Murano, Laura Ronchetti, Paolo Signoretti, Lisa Volpe, Domenico Palladino
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Recent European policies have been set ambitious targets aimed at significantly reducing CO2 emissions by 2030, with a long-term vision of transforming existing buildings into Zero-Emissions Buildings (ZEmB) by 2050. This vision represents a key point for the energy transition as the whole building stock currently accounts for 36% of total energy consumption across the Europe, mainly due to their poor energy performance. The challenge towards Zero-Emissions Buildings is particularly felt in Italy, where a significant number of buildings with historical significance or situated within protected/constrained areas can be found. Furthermore, an estimated 70% of the national building stock are built before 1976, indicating a widespread issue of poor energy performance. Addressing the energy ineƯiciency of these buildings is crucial to refining a comprehensive energy renovation approach aimed at facilitating their energy transition. In this framework the current study focuses on analysing a challenging complex of buildings to be totally restored through significant energy renovation interventions. The goal is to recover these disused buildings situated in a significant archaeological zone of Rome, contributing to the restoration and reintegration of this historically valuable site, while also oƯering insights useful for achieving zeroemission requirements for buildings within such contexts. In pursuit of meeting the stringent zero-emission requirements, a comprehensive study was carried out to assess the complex of buildings, envisioning substantial renovation measures on building envelope and plant systems and incorporating renewable energy system solutions, always respecting and preserving the historic site. An energy audit of the complex of buildings was performed to define the actual energy consumption for each energy service by adopting the hourly calculation methods. Subsequently, significant energy renovation interventions on both building envelope and mechanical systems have been examined respecting the historical value and preservation of site. These retrofit strategies have been investigated with threefold aims: 1) to recover the existing buildings ensuring the energy eƯiciency of the whole complex of buildings, 2) to explore which solutions have allowed achieving and facilitating the ZEmB status, 3) to balance the energy transition requirements with the sustainable aspect in order to preserve the historic value of the buildings and site. This study has pointed out the potentiality and the technical challenges associated with implementing renovation solutions for such buildings, representing one of the first attempt towards realizing this ambitious target for this type of building.Keywords: energy conservation and transition, complex of buildings in historic site, zero-emission buildings, energy efficiency recovery
Procedia PDF Downloads 773745 MicroRNA Profiling Reveals Novel Circulating Biomarkers in Acute Phase of Myocardial Infarction
Authors: A. Maciejak, M. Kiliszek, G. Opolski, D. Tulacz, A. Segiet, K. Matlak, S. Dobrzycki, G. Sygitowicz, B. Burzynska, M. Gora
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Introduction and aims: Acute myocardial infarction (AMI) is one of the most severe cardiovascular diseases affecting millions of patients each year worldwide. An early and accurate diagnosis of AMI is essential for optimal treatment. Therefore, new approaches that can complement and improve current strategies for AMI diagnosis are urgently needed. Recent studies have revealed the presence of stable circulating myocardial-derived microRNAs (miRNAs) in human peripheral blood, suggesting that such miRNAs could serve as potential biomarkers of infarction. The present study aimed to identify differentially expressed circulating miRNAs in ST-segment elevation myocardial infarction (STEMI) patients. Materials and methods: miRNA expression profile analysis was performed using Exiqon Serum/Plasma Focus microRNA PCR panel in plasma samples of n=16 patients on the first day of AMI (admission) and in samples from the same patients collected six months after AMI. Selected miRNAs were validated by RT-qPCR using serum samples from an independent set of n=14 AMI patients. Results: The profiling study identified 46 species of plasma miRNAs that were differentially expressed (p < 0.05) on admission compared to six months after AMI. The validation in the independent group of patients confirmed that miR-133b and miR-22-5p were significantly up-regulated upon AMI. Conclusions: Our results suggest that miRNA expression profiling provides better understanding of the changes that occur in the acute phase of MI in the myocardium and could be useful in determination of the potential role of extracellular miRNAs as paracrine signaling molecules. miR-22-5p represents a novel promising biomarker for the diagnosis of acute myocardial infarction.Keywords: acute myocardial infarction, circulating microRNAs, microRNA expression profiling, miR-22-5p
Procedia PDF Downloads 3323744 Following the Modulation of Transcriptional Activity of Genes by Chromatin Modifications during the Cell Cycle in Living Cells
Authors: Sharon Yunger, Liat Altman, Yuval Garini, Yaron Shav-Tal
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Understanding the dynamics of transcription in living cells has improved since the development of quantitative fluorescence-based imaging techniques. We established a method for following transcription from a single copy gene in living cells. A gene tagged with MS2 repeats, used for mRNA tagging, in its 3' UTR was integrated into a single genomic locus. The actively transcribing gene was detected and analyzed by fluorescence in situ hybridization (FISH) and live-cell imaging. Several cell clones were created that differed in the promoter regulating the gene. Thus, comparative analysis could be obtained without the risk of different position effects at each integration site. Cells in S/G2 phases could be detected exhibiting two adjacent transcription sites on sister chromatids. A sharp reduction in the transcription levels was observed as cells progressed along the cell cycle. We hypothesized that a change in chromatin structure acts as a general mechanism during the cell cycle leading to down-regulation in the activity of some genes. We addressed this question by treating the cells with chromatin decondensing agents. Quantifying and imaging the treated cells suggests that chromatin structure plays a role both in regulating transcriptional levels along the cell cycle, as well as in limiting an active gene from reaching its maximum transcription potential at any given time. These results contribute to understanding the role of chromatin as a regulator of gene expression.Keywords: cell cycle, living cells, nucleus, transcription
Procedia PDF Downloads 3133743 Analysis of the Degradation of the I-V Curve of the PV Module in a Harsh Environment: Estimation of the Site-Specific Factor (Installation Area)
Authors: Maibigue Nanglet, Arafat Ousman Béchir, Mahamat Hassan Béchir
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The economy of Central African countries is growing very fast, and the demand for energy is increasing every day. As a result, insufficient power generation is one of the major problems slowing down development. This paper explores the factors of degradation of the I-V curve of the PV Generator (GPV) in harsh environments, taking the case of two locals: Mongo and Abeche. Its objective is to quantify the voltage leaks due to the different GPV installation areas; after using the Newton-Raphson numerical method of the solar cell, a survey of several experimental measurement points was made. The results of the simulation in MATLAB/Simulink show a relative power loss factor of 11.8765% on the GPVs installed in Mongo and 8.5463% on those installed in Abeche; these results allow us to say that the supports on which the modules are installed have an average impact of 10.2114% on their efficiency.Keywords: calculation, degradation, site, GPV, severe environment
Procedia PDF Downloads 403742 Production of Insulin Analogue SCI-57 by Transient Expression in Nicotiana benthamiana
Authors: Adriana Muñoz-Talavera, Ana Rosa Rincón-Sánchez, Abraham Escobedo-Moratilla, María Cristina Islas-Carbajal, Miguel Ángel Gómez-Lim
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The highest rates of diabetes incidence and prevalence worldwide will increase the number of diabetic patients requiring insulin or insulin analogues. Then, current production systems would not be sufficient to meet the future market demands. Therefore, developing efficient expression systems for insulin and insulin analogues are needed. In addition, insulin analogues with better pharmacokinetics and pharmacodynamics properties and without mitogenic potential will be required. SCI-57 (single chain insulin-57) is an insulin analogue having 10 times greater affinity to the insulin receptor, higher resistance to thermal degradation than insulin, native mitogenicity and biological effect. Plants as expression platforms have been used to produce recombinant proteins because of their advantages such as cost-effectiveness, posttranslational modifications, absence of human pathogens and high quality. Immunoglobulin production with a yield of 50% has been achieved by transient expression in Nicotiana benthamiana (Nb). The aim of this study is to produce SCI-57 by transient expression in Nb. Methodology: DNA sequence encoding SCI-57 was cloned in pICH31070. This construction was introduced into Agrobacterium tumefaciens by electroporation. The resulting strain was used to infiltrate leaves of Nb. In order to isolate SCI-57, leaves from transformed plants were incubated 3 hours with the extraction buffer therefore filtrated to remove solid material. The resultant protein solution was subjected to anion exchange chromatography on an FPLC system and ultrafiltration to purify SCI-57. Detection of SCI-57 was made by electrophoresis pattern (SDS-PAGE). Protein band was digested with trypsin and the peptides were analyzed by Liquid chromatography tandem-mass spectrometry (LC-MS/MS). A purified protein sample (20µM) was analyzed by ESI-Q-TOF-MS to obtain the ionization pattern and the exact molecular weight determination. Chromatography pattern and impurities detection were performed using RP-HPLC using recombinant insulin as standard. The identity of the SCI-57 was confirmed by anti-insulin ELISA. The total soluble protein concentration was quantified by Bradford assay. Results: The expression cassette was verified by restriction mapping (5393 bp fragment). The SDS-PAGE of crude leaf extract (CLE) of transformed plants, revealed a protein of about 6.4 kDa, non-present in CLE of untransformed plants. The LC-MS/MS results displayed one peptide with a high score that matches SCI-57 amino acid sequence in the sample, confirming the identity of SCI-57. From the purified SCI-57 sample (PSCI-57) the most intense charge state was 1069 m/z (+6) on the displayed ionization pattern corresponding to the molecular weight of SCI-57 (6412.6554 Da). The RP-HPLC of the PSCI-57 shows the presence of a peak with similar retention time (rt) and UV spectroscopic profile to the insulin standard (SCI-57 rt=12.96 and insulin rt=12.70 min). The collected SCI-57 peak had ELISA signal. The total protein amount in CLE from transformed plants was higher compared to untransformed plants. Conclusions: Our results suggest the feasibility to produce insulin analogue SCI-57 by transient expression in Nicotiana benthamiana. Further work is being undertaken to evaluate the biological activity by glucose uptake by insulin-sensitive and insulin-resistant murine and human cultured adipocytes.Keywords: insulin analogue, mass spectrometry, Nicotiana benthamiana, transient expression
Procedia PDF Downloads 3513741 IL-33 Production in Murine Macrophages via PGE2-E Prostanoid Receptor 2/4 Signaling
Authors: Sachin K. Samuchiwal, Barbara Balestrieri, Amanda Paskavitz, Hannah Raff, Joshua A. Boyce
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IL-33, a recently discovered member of the IL-1 cytokine family, binds to the TLR/IL1R super family receptor ST2 and induces type 2 immune responses. IL-33 is constitutively expressed in structural cells at barrier sites such as skin, lung, and intestine, and also inducibly expressed by hematopoietic cells including macrophages. Stimulation of macrophages by Lipopolysaccharide (LPS) can induce de novo IL-33 expression, and also causes the production of prostaglandin-E2 (PGE2) via cyclooxygenase (COX)-2 and microsomal PGE2 synthase-1 (mPGES-1). Because PGE2 can regulate macrophage functions through both autocrine and paracrine mechanisms, the potential interplay of endogenous PGE2 on IL-33 production was explored. Bone-marrow derived murine macrophages (bmMF) that lack either mPGES-1 or EP2 receptor expression were stimulated with LPS in the absence or presence of exogenous PGE2 along with pharmacological agonists and antagonists. The study results demonstrate that endogenous PGE2 markedly enhances LPS-induced IL-33 production by bmMFs via EP2 receptors. Moreover, exogenous PGE2 can amplify LPS-induced IL-33 expression dominantly by EP2 and partly by EP4 receptors by a pathway involving cAMP and exchange protein activated by cAMP (EPAC), but not protein kinase A (PKA). Though both IL-33 production and PGE2 generation in response to LPS require activation of both p38 MAPK and NF-κB, PGE2 did not influence this activation. In conclusion, it is demonstrated that endogenous PGE2 signaling through EP2 and EP4 receptors is a prerequisite for LPS-induced IL-33 production in bmMFs and the underlying cAMP mediated pathway involves EPAC. Since IL-33 is a critical pro-inflammatory cytokine in various pathological disorders, this PGE2-EP2/EP4-cAMP mediated pathway can be exploited to intervene in IL-33 driven pathologies.Keywords: bone marrow macrophages, EPAC, IL-33, PGE2
Procedia PDF Downloads 1913740 Juvenile Fish Associated with Pondweed and Charophyte Habitat: A Case Study Using Upgraded Pop-up Net in the Estuarine Part of the Curonian Lagoon
Authors: M. Bučas, A. Skersonas, E. Ivanauskas, J. Lesutienė, N. Nika, G. Srėbalienė, E. Tiškus, J. Gintauskas, A.Šaškov, G. Martin
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Submerged vegetation enhances heterogeneity of sublittoral habitats; therefore, macrophyte stands are essential elements of aquatic ecosystems to maintain a diverse fish fauna. Fish-habitat relations have been extensively studied in streams and coastal waters, but in lakes and estuaries are still underestimated. The aim of this study is to assess temporal (diurnal and seasonal) patterns of fish juvenile assemblages associated with common submerged macrophyte habitats, which have significantly spread during the recent decade in the upper littoral part of the Curonian Lagoon. The assessment was performed by means of an upgraded pop-up net approach resulting in much precise sampling versus other techniques. The optimal number of samples (i.e., pop-up nets) required to cover>80% of the total number of fish species depended on the time of the day in both study sites: at least 7and 9 nets in the evening (18-24 pm) in the Southern and Northern study sites, respectively. In total, 14 fish species were recorded, where perch and roach dominated (respectively 48% and 24%). From multivariate analysis, water salinity and seasonality (temperature or sampling month) were primary factors determining fish assemblage composition. The southern littoral area, less affected by brackish water conditions, hosted a higher number of species (13) than in the Northern site (8). In the latter site, brackish water tolerant species (three-spined and nine-spined sticklebacks, spiny loach, roach, and round goby) were more abundant than in the Southern site. Perch and ruffe dominated in the Southern site. Spiny loach and nine-spined stickleback were more frequent in September, while ruffe, perch, and roach occurred more in July. The diel dynamics of the common species such as perch, roach, and ruffe followed the general pattern, but it was species specific and depended on the study site, habitat, and month. The species composition between macrophyte habitats did not significantly differ; however, it differed from the results obtained in 2005 at both study sites indicating the importance of expanded charophyte stands during the last decade in the littoral zone.Keywords: diel dynamics, charophytes, pondweeds, herbivorous and benthivorous fishes, littoral, nursery habitat, shelter
Procedia PDF Downloads 1893739 Biofilm Formation Due to the Proteome Changes Of Enterococcus Faecium in Response to Sub-Mic of Gentamicin
Authors: Amin Abbasi, Mahdi Asghari Ozma
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Background and Objective:Enterococcus faecium is a normal flora of the human gastrointestinal tract that causes infection in the host body under conditions such as biofilm formation, in which the use of antibiotics causes changes in these pathogenic mechanisms. In this study, we aimed to evaluate comprehensively the changes in E.faecium when exposed to sub-MIC of the gentamicin,especiallythe biofilm formation rate. Materials and Methods: For this study, the keywords "Enterococcus faecium ", "Biofilm", and "Gentamicin" in the databases PubMed, Google Scholar, Sid, and MagIran between 2015 and 2021 were searched, and 14 articles were chosen, studied, and analyzed. Results: Gentamicin significantly had increased biofilm formation in most of the isolates in the studies. Increased expression of the genes (efaA and esp) and proteins involved in biofilm formation and decreased expression of the genes (gelE and cylA) involved in spreading and proteins involved in metabolism and cell division in E.faecium were the most significant cause of the biofilm formation, which were increased in sub-MIC gentamicin-treated situation. Conclusion: Inadequate use of gentamicin intensify biofilm formation of E.faecium, which can make the treatment of infections caused by this bacterium difficult.Keywords: biofilm, enterococcus faecium, gentamicin, proteome
Procedia PDF Downloads 1113738 Detection of Cytotoxicity of Green Synthesized Silver, Gold, and Silver/Gold Bimetallic on Baby Hamster Kidney-21 Cells Using MTT Assay
Authors: Naila Sher, Mushtaq Ahmed, Nadia Mushtaq, Rahmat Ali Khan
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In cancer therapy, nanoparticles (NPs) shall be applied possibly by inoculation in the veins of humans. This action will connect them with white (WBCs) and red blood cells (RBCs) in the bloodstream before they reach their main targeted cancer cells. However, possible effects of silver, gold, and silver/gold bimetallic NPs (Ag, Au, and Ag/Au BNPs) on baby hamster kidney-21 (BHK-21) cells were studied by MTT assay. Here, Ag, Au, and their Ag/Au BNPs (bimetallic nanoparticles) were synthesized by using Hippeastrum hybridum (HH) extract. These NPs were characterized by UV-visible spectroscopy, FT-IR, XRD, and EDX, and SEM analysis. XRD analysis conferring the crystal structure with an average size of 13.3, 10.72, and 8.34nm of Ag, Au, and Ag/Au BNPs, respectively. SEM showed that Ag, Au, and Ag/Au BNPs had irregular morphologies, with nano measures calculated sizes of 40, 30, and 20 nm respectively. EDX spectrometers confirmed the presence of elemental Ag signal of the AgNPs with 22.75%, Au signal of the AuNPs with 48.08%, Ag signal with 12%, and Au signal with 38.26% of the Ag/Au BNPs. The BHK-21cells were incubated in the existence of doxorubicin, plant extract, Ag, Au, and Ag/Au BNPs. The cytotoxic effects could be observed in a dose-dependent mode; doxorubicin and Ag/Au BNPs were more toxic than plant extract, Ag, and Au NPs. It is demonstrated that NPs interact with BHK-21cells and significantly reduce cell viability in a concentration-dependent manner. However, to reduce the potential threats of NPs further studies are recommended.Keywords: hippeastrum hybridum, nanoparticle, BHK-21cells
Procedia PDF Downloads 1343737 Enhanced Wound Healing Efficacy of Cordycepin-Melittin Nanoconjugate in Excised Wounds of Diabetic Rats
Authors: Mohammed Flaih Alotaibi, Rasheed Ahemad Shaik, Mohammed Z. Nasrullah
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Diabetic foot ulcers are the foremost global healthcare burden. Hyperglycemia in diabetics is incriminating in impeding wound healing and it can allow for more severe medical issues. The study was intended to establish a nanoconjugate of cordycepin-melittin (COR-MEL) and evaluate its healing effects in wounded diabetic rats. Diabetes induced by injecting streptozotocin intraperitoneally (50 mg/kg, body weight). Therefore, animals were classified into various groups; diabetic untreated, vehicle-treated, COR alone, MEL alone, and COR-MEL nanoconjugate treated, respectively. Animals with diabetes were exposed to excision and treated with Vehicle, COR, MEL, or COR-MEL nanoconjugate topically. After 14 days, the wounded skin was sliced and subjected to histological and biochemical assessments. The formulated nanoconjugate has a particle size of 253.5± 17.4 nm by a polydispersity index of 0.36 ± 0.05, and a zeta potential of 1.72 ± 0.3 mV. The study demonstrated an accelerated wound contraction in COR-MEL-treated diabetic rats, which was further validated by histological analysis. The nanoconjugate further exhibited antioxidant activities by inhibiting the accumulation of malondialdehyde and exhaustion of superoxide dismutase and glutathione peroxidase enzymatic activities. The nanoconjugate further demonstrated an enhanced anti-inflammatory activity by retarding the expression of proinflammatory cytokines (IL-6 and TNF-α). Additionally, the nanoconjugate exhibits a strong expression of growth factors (TGF-β1, VEGF-A, and PDGFR-β), indicating enrichment of proliferation. Likewise, nanoconjugate increased the concentration of hydroxyproline as well as the mRNA expression of collagen, type I, alpha 1. Thus, it is concluded that the nanoconjugate possesses a potent wound-healing activity in diabetic rats via antioxidant, anti-inflammatory, and pro-angiogenetic mechanisms.Keywords: diabetic wounds, cordycepin, melittin, nanoconjugate, wound healing
Procedia PDF Downloads 813736 Rodriguez Diego, Del Valle Martin, Hargreaves Matias, Riveros Jose Luis
Authors: Nathainail Bashir, Neil Anderson
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The objective of this study site was to investigate the current state of the practice with regards to karst detection methods and recommend the best method and pattern of arrays to acquire the desire results. Proper site investigation in karst prone regions is extremely valuable in determining the location of possible voids. Two geophysical techniques were employed: multichannel analysis of surface waves (MASW) and electric resistivity tomography (ERT).The MASW data was acquired at each test location using different array lengths and different array orientations (to increase the probability of getting interpretable data in karst terrain). The ERT data were acquired using a dipole-dipole array consisting of 168 electrodes. The MASW data was interpreted (re: estimated depth to physical top of rock) and used to constrain and verify the interpretation of the ERT data. The ERT data indicates poorer quality MASW data were acquired in areas where there was significant local variation in the depth to top of rock.Keywords: dipole-dipole, ERT, Karst terrains, MASW
Procedia PDF Downloads 3153735 Efficient L-Xylulose Production Using Whole-Cell Biocatalyst With NAD+ Regeneration System Through Co-Expression of Xylitol Dehydrogenase and NADH Oxidase in Escherichia Coli
Authors: Mesfin Angaw Tesfay
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L-Xylulose is a potentially valuable rare sugar used as starting material for antiviral and anticancer drug development in pharmaceutical industries. L-Xylulose exist in a very low concentration in nature and have to be synthesized from cheap starting materials such as xylitol through biotechnological approaches. In this study, cofactor engineering and deep eutectic solvent were applied to improve the efficiency of L-xylulose production from xylitol. A water-forming NAD+ regeneration enzyme (NADH oxidase) from Streptococcus mutans ATCC 25175 was introduced into E. coli with xylitol-4-dehydrogenase (XDH) of Pantoea ananatis resulting in recombinant cells harboring the vector pETDuet-xdh-SmNox. Further, three deep eutectic solvents (DES) including, Choline chloride/glycerol (ChCl/G), Choline chloride/urea (ChCl/U), and Choline chloride/ethylene glycol (ChCl/EG) have been employed to facilitate the conversion efficiency of L-xylulose from xylitol. The co-expression system exhibited optimal activity at a temperature of 37 ℃ and pH 8.5, and the addition of Mg2+ enhanced the catalytic activity by 1.19-fold. Co-expression of NADH oxidase with XDH enzyme resulted in increased L-xylulose concentration and productivity from xylitol as well as the intracellular NAD+ concentration. Two of the DES used (ChCl/U and ChCl/EG) show positive effects on product yield and the ChCl/G has inhibiting effects. The optimum concentration of ChCl/U was 2.5%, which increased the L-xylulose yields compared to the control without DES. In a 1 L fermenter the final concentration and productivity of L-xylulose from 50 g/L of xylitol reached 48.45 g/L, and 2.42 g/L.h respectively, which was the highest report. Overall, this study is a suitable approach for large-scale production of L-xylulose from xylitol using the engineered E. coli cell.Keywords: Xylitol-4-dehydrogenase, NADH oxidase, L-xylulose, Xylitol, Coexpression, DESs
Procedia PDF Downloads 273734 Spectral Assessing of Topographic Effects on Seismic Behavior of Trapezoidal Hill
Authors: M. Amelsakhi, A. Sohrabi-Bidar, A. Shareghi
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One of the most important issues about the structural damages caused by earthquake is the evaluating of the spectral response of the site on which the construction is built. This fact has demonstrated during many earlier earthquakes and many researchers’ reports have concerned with it. According to these reports, features of the site materials and geometry of the ground surface are considered the main factors. This study concentrates on the specific form of topographies like hills. Assessing of spectral responses of different points on the hills and beside demonstrates considerable differences between 1D and 2D methods of geotechnical analyses. A general trend of amplifications on the top of the hills and de-amplifications near the toe of the hills has been appeared within the acceleration, velocity and displacement response spectrums of horizontal motion. Evaluating of spectral responses of different sizes of the hills revealed that as much as the hill-size enlarges differences between spectral responses of 1D and 2D analyses transfers to longer range of periods and becomes wider.Keywords: topography effect, amplification ratio, response spectrum, earth resources engineering
Procedia PDF Downloads 2393733 Basal Cell Carcinoma Excision Intraoperative Frozen Section for Tumor Clearance and Reconstructive Surgery: A Prospective Open Label Interventional Study
Authors: Moizza Tahir, Uzma Bashir, Aisha Akhtar, Zainab Ansari, Sameen Ansari, Muhammad Ali Tahir
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Cancer burden has globally increased. Among cutaneous cancers basal cell carcinoma constitute vast majority of skin cancer. There is need for appropriate diagnostic, therapeutic and prognostic significance evaluation for skin cancers Present study report intraoperative frozen section (FS) histopathological clearance for excision of BCC in a tertiary care center and find the frequency of involvement of surgical margin with reference to anatomical site, with size and surgical technique. It was prospective open label interventional study conducted at Dermatology department of tertiary care hospital Rawalpindi Pakistan in lais on with histopathology department from January 2023 to April 2024. Total of thirty-six (n = 36) patients between age 45-80 years with basal cell carcinoma of 10-20mm on face were included following inclusion exclusion criteria by purposive sampling technique. Informed consent was taken. Surgical excision was performed and intraoperative frozen section histopathology clearance of tumor margin was taken from histopathologist on telephone. Surgical reconstruction was done. Final Histopathology report was reexamined on day 10th for margin and depth clearance. Descriptive statistics were calculated for age, gender, sun exposure, reconstructive technique, anatomical site, and tumor free margin report on frozen section analysis. Chi square test was employed for statistical significance of involvement of surgical margin with reference to anatomical site, size and decision on reconstructive surgical technique, p value of <0.05 was considered significant. Total of 36 patients of BCC were enrolled, males 12 (33.3%) and females were 24 (66.6%). Age ranged from 45 year to 80 year mean of 58.36 ±SD7.8. Size of BCC ranged from 10mm to 35mm mean of 25mm ±SD 0.63. Morphology was nodular 18 (50%), superficial spreading 11(30.6%), morphoeic 1 (2.8%) and ulcerative in 6(16.7%) cases. Intraoperative frozen section for histopathological margin clearance with 2-3 mm safety margin and surgical technique has p-value0.51, for anatomical site p value 0.24 and size p-0.84. Intraoperative frozen section (FS) histopathological clearance for BCC face with 2-3mm safety margin with reference to reconstructive technique, anatomical site and size of BCC were insignificant.Keywords: basal cell carcinoma, tumor free amrgin, basal cell carcinoma and frozen section, safety margin
Procedia PDF Downloads 573732 Impact of Tuberculosis Co-infection on Cytokine Expression in HIV-Infected Individuals
Authors: M. Nosik, I. Rymanova, N. Adamovich, S. Sevostyanihin, K. Ryzhov, Y. Kuimova, A. Kravtchenko, N. Sergeeva, A. Sobkin
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HIV and Tuberculosis (TB) infections each speed the other's progress. HIV-infection increases the risk of TB disease. At the same time, TB infection is associated with clinical progression of HIV-infection. HIV+TB co-infected patients are also at higher risk of acquiring new opportunistic infections. An important feature of disease progression and clinical outcome is the innate and acquired immune responses. HIV and TB, however, have a spectrum of dysfunctions of the immune response. As cytokines play a crucial role in the immunopathology of both infections, it is important to study immune interactions in patients with dual infection HIV+TB. Plasma levels of proinflammatory cytokines IL-2, IFN-γ and immunoregulating cytokines IL-4, IL-10 were evaluated in 75 patients with dual infection HIV+TB, 58 patients with HIV monoinfection and 50 patients with TB monoinfection who were previously naïve for HAART. The decreased levels of IL-2, IFN-γ, IL-4 and IL-10 were observed in patients with dual infection HIV+TB in comparison with patients who had only HIV or TB which means the profound suppression of Th1 and Th2 cytokine secretion. Thus, those cytokines could possibly serve as immunological markers of progression of HIV-infection in patients with TB.Keywords: HIV, tuberculosis (TB), HIV associated with TB, Th1/ Th2 cytokine expression
Procedia PDF Downloads 3653731 Enzyme Redesign: From Metal-Dependent to Metal-Independent, a Symphony Orchestra without Concertmasters
Authors: Li Na Zhao, Arieh Warshel
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The design of enzymes is an extremely challenging task, and this is also true for metalloenzymes. In the case of naturally evolved enzymes, one may consider the active site residues as the musicians in the enzyme orchestra, while the metal can be considered as their concertmaster. Together they catalyze reactions as if they performed a masterpiece written by nature. The Lactonase can be thought as a member of the amidohydrolase family, with two concertmasters, Fe and Zn, at its active site. It catalyzes the quorum sensing signal- N-acyl homoserine lactones (AHLs or N-AHLs)- by hydrolyzing the lactone ring. This process, known as quorum quenching, provides a strategy in the treatment of infectious diseases without introducing selection pressure. However, the activity of lactonase is metal-dependent, and this dependence hampers the clinic usage. In our study, we use the empirical valence bond (EVB) approach to evaluate the catalytic contributions decomposing them to electrostatic and other components.Keywords: enzyme redesign, empirical valence bond, lactonase, quorum quenching
Procedia PDF Downloads 2543730 Inferring Thimlich Ohinga Gender Identity Through Ethnoarchaeological Analysis
Authors: David Maina Muthegethi
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The Victoria Basin is associated with gateway for migration to Southern part of Africa. Different communities migrated through the region including the Bantus and Nilotic communities that occupy present day Kenya and Tanzania. A distinct culture of dry-stone technology emerged around 15th century current era, a period associated with peopling of the western Kenya region. One of the biggest dry-stone walls enclosure is Thimlich Ohinga archaeological site. The site was constructed around fourteenth century current era. Architectural design was oval shaped stone structures that were around 4 meters and 2 meters in length and width respectively. The main subsistence strategies of the community that was crop faming, pastoralism, fishing, hunting and gathering. This paper attempts to examine gender dynamics of Thimlich Ohinga society. At that end, attempts are made to infer gender roles as manifested in archaeological record. Therefore, the study entails examination of material evidence excavated from the site. Also, ethnoarchaeological study of contemporary Luo community was undertaken in order to make inferences and analogies concerning gender roles of Thimlich Ohinga society. Overall, the study involved examination of cultural materials excavated from Thimlich Ohinga, extensive survey of the site and ethnography of Luo community. In total, an extensive survey and interviews of 20 households was undertaken in South Kanyamkango ward, Migori County in Western Kenya. The key findings point out that Thimlich Ohinga gender identities were expressed in material forms through architecture, usage of spaces, subsistence strategies, dietary patterns and household organization. Also, gender as social identity was dynamic and responsive to diversification of subsistence strategies and intensification of regional trade as documented in contemporary Luo community. The paper reiterates importance of ethnoarchaeological methods in reconstruction of past social organization as manifested in material record.Keywords: ethnoarchaeological, gender, subsistence patterns, Thimlich Ohinga
Procedia PDF Downloads 773729 Signs and Symbols of a Modern Sufi Ceremony in Istanbul
Authors: Antonio De Lisa
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The aim of this contribution is to describe a Sufi ceremony personally observed and recorded in Istanbul. It is part of research on the semiotics of religion that has been going on for some time. Over the centuries, Sufism has produced remarkable literature, elaborated mainly in the context of Arabic literature and Persian literature, but has also found expression in many other languages (Turkish, Indian, Malayo-Indonesian, etc.). Among the genres cultivated are: devotional books (prayers, meditation, spiritual exercises, etc.); the hagiographic texts, containing the biographies and sentences of the best-known Sufis; the texts that illustrate the abodes or stations of the spiritual path; finally, the theoretical treatises on various topics, often of an apologetic nature. Another typical expression of Sufism is in the literature in verse, which includes first-rate poets of both Arabic (Ibn al-Farid, Ibn 'Arabi) and Persian (Farid al-Din al-Attar, Rumi, Hafez, Gohar Shahi) expression. To observe an authentic Sufi ceremony, we contacted the Silivrikapı Mevlana Cultural Center in Istanbul. The ceremony we will talk about is a characteristic of the brotherhood called Mevleviyye (Mawlawiyya), so called from Mawlānā ('our lord', in Turkish: Mevlānā) Jalāl al-Dīn Muḥammad Rūmī (13th century) better known as the brotherhood of the Whirling Dervishes, founded in Konya. The characteristic of the order is to pray about one's love for Allah with music and dance in a ritual that has the name of Semà. The dance is a swirling movement in a group, representing a mystical journey of the spiritual ascent of man through the mind and love for the 'perfect'. Turning towards the truth, the follower of the group ascends towards love, passing beyond the World and Nothingness. After asceticism, he returns to the common world to communicate his own experience.Keywords: semiotics of religion, sufi, rumi, islamic philosophy
Procedia PDF Downloads 923728 Suitability Assessment of Water Harvesting and Land Restoration in Catchment Comprising Abandoned Quarry Site in Addis Ababa, Ethiopia
Authors: Rahel Birhanu Kassaye, Ralf Otterpohl, Kumelachew Yeshitila
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Water resource management and land degradation are among the critical issues threatening the suitable livability of many cities in developing countries such as Ethiopia. Rapid expansion of urban areas and fast growing population has increased the pressure on water security. On the other hand, the large transformation of natural green cover and agricultural land loss to settlement and industrial activities such as quarrying is contributing to environmental concerns. Integrated water harvesting is considered to play a crucial role in terms of providing alternative water source to insure water security and helping to improve soil condition, agricultural productivity and regeneration of ecosystem. Moreover, it helps to control stormwater runoff, thus reducing flood risks and pollution, thereby improving the quality of receiving water bodies and the health of inhabitants. The aim of this research was to investigate the potential of applying integrated water harvesting approaches as a provision for water source and enabling land restoration in Jemo river catchment consisting of abandoned quarry site adjacent to a settlement area that is facing serious water shortage in western hilly part of Addis Ababa city, Ethiopia. The abandoned quarry site, apart from its contribution to the loss of aesthetics, has resulted in poor water infiltration and increase in stormwater runoff leading to land degradation and flooding in the downstream. Application of GIS and multi-criteria based analysis are used for the assessment of potential water harvesting technologies considering the technology features and site characteristics of the case study area. Biophysical parameters including precipitation, surrounding land use, surface gradient, soil characteristics and geological aspects are used as site characteristic indicators and water harvesting technologies including retention pond, check dam, agro-forestation employing contour trench system were considered for evaluation with technical and socio-economic factors used as parameters in the assessment. The assessment results indicate the different suitability potential among the analyzed water harvesting and restoration techniques with respect to the abandoned quarry site characteristics. Application of agro-forestation with contour trench system with the revegetation of indigenous plants is found to be the most suitable option for reclamation and restoration of the quarry site. Successful application of the selected technologies and strategies for water harvesting and restoration is considered to play a significant role to provide additional water source, maintain good water quality, increase agricultural productivity at urban peri-urban interface scale and improve biodiversity in the catchment. The results of the study provide guideline for decision makers and contribute to the integration of decentralized water harvesting and restoration techniques in the water management and planning of the case study area.Keywords: abandoned quarry site, land reclamation and restoration, multi-criteria assessment, water harvesting
Procedia PDF Downloads 2173727 Difficulties in the Emotional Processing of Intimate Partner Violence Perpetrators
Authors: Javier Comes Fayos, Isabel RodríGuez Moreno, Sara Bressanutti, Marisol Lila, Angel Romero MartíNez, Luis Moya Albiol
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Given the great impact produced by gender-based violence, its comprehensive approach seems essential. Consequently, research has focused on risk factors for violent behaviour, linking various psychosocial variables, as well as cognitive and neuropsychological deficits with the aggressors. However, studies on affective processing are scarce, so the present study investigates possible emotional alterations in men convicted of gender violence. The participants were 51 aggressors, who attended the CONTEXTO program with sentences of less than two years, and 47 men with no history of violence. The sample did not differ in age, socioeconomic level, education, or alcohol and other substances consumption. Anger, alexithymia and facial recognition of other people´s emotions were assessed through the State-Trait Anger Expression Inventory (STAXI-2), the Toronto Alexithymia Scale (TAS-20) and Reading the mind in the eyes (REM), respectively. Men convicted of gender-based violence showed higher scores on the anger trait and temperament dimensions, as well as on the anger expression index. They also scored higher on alexithymia and in the identification and emotional expression subscales. In addition, they showed greater difficulties in the facial recognition of emotions by having a lower score in the REM. These results seem to show difficulties in different affective areas in men condemned for gender violence. The deficits are reflected in greater difficulty in identifying and expressing emotions, in processing anger and in recognizing the emotions of others. All these difficulties have been related to the use of violent behavior. Consequently, it is essential and necessary to include emotional regulation in intervention programs for men who have been convicted of gender-based violence.Keywords: alexithymia, anger, emotional processing, emotional recognition, empathy, intimate partner violence
Procedia PDF Downloads 2023726 The Role of Deformation Strain and Annealing Temperature on Grain Boundary Engineering and Texture Evolution of Haynes 230
Authors: Mohsen Sanayei, Jerzy Szpunar
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The present study investigates the effects of deformation strain and annealing temperature on the formation of twin boundaries, deformation and recrystallization texture evolution and grain boundary networks and connectivity. The resulting microstructures were characterized using Electron Backscatter Diffraction (EBSD) and X-Ray Diffraction (XRD) both immediately following small amount of deformation and after short time annealing at high temperature to correlate the micro and macro texture evolution of these alloys. Furthermore, this study showed that the process of grain boundary engineering, consisting cycles of deformation and annealing, is found to substantially reduce the mass and size of random boundaries and increase the proportion of low Coincidence Site Lattice (CSL) grain boundaries.Keywords: coincidence site lattice, grain boundary engineering, electron backscatter diffraction, texture, x-ray diffraction
Procedia PDF Downloads 3143725 The Mediating Role of Early Maladaptive Schemas in the Relationship between Attachment and Trait Anger and Anger Expression
Authors: Ayperi̇ Haspolat Özcan, Meltem Anafarta Şendağ
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This study aimed to establish a model in the light of current approaches for understanding the mediating role of early maladaptive schemas in the relationship between attachment and anger. Accordingly, the proposed mediation model was tested by mediation with bootstrapping technique, considering gender and attachment figure differences. The university students (N= 444) with ages ranging from 17 to 28 participated in the study. Participants filled out Parental and Peer Attachment Scale Short Form, Young Schema Questionnaire - Short Form 3, Trait Anger and Anger Expression Scales. The mediating role of early maladaptive schemas (impaired autonomy, disconnection and rejection, unrelenting standards, other-directedness, and impaired limits) in the relationship between attachment (mother and father) and anger aspects (trait anger, anger in, anger out and anger control) were found to be significant for both male and female participants. Separate mediation analyses for both genders and different attachment figures have also drawn attention to noticeable differences in the results. Specifically, for females, various paths were discovered in predicting various aspects of anger (anger in, anger out, anger control, and trait anger). On the other hand, for males only anger directed inwards was found to be predicted by any source of attachment through disconnection and rejection schema only. These obvious gender differences in understanding the mechanism of anger are discussed in the light of cultural gender roles and the social acceptance of anger in males. In the area of application, the study of various aspects of anger with particular attention to attachment and early maladaptive schemas as well as the importance of distinguishing the gender differences are emphasized as important points.Keywords: anger expression, attachment, early maladaptive schemas, trait anger
Procedia PDF Downloads 2883724 A Case Study: Remediation of Abandoned Mines for Residential Development
Authors: Issa S. Oweis, Gary Gartenberg, Luma J. Oweis
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The site for a residential apartment building overlies an abandoned iron mine in granitic gneiss in northern New Jersey. The mine stope is about 137 m (450 long) and dipping over 344m (800 feet) at 450 to 500. As the building footprint straddles, the mine site needed remediation. The remediation scheme consisted of compaction grouting a minimum 10 m (30 ft.) depth of the mine stope in rock to establish a buttress for the hanging wall and allow support of the building foundation. The rock strength parameters (friction and cohesion) were established based on Hoek Geologic Strength Index (GSI). The derived strength parameters were used in the wedge analysis to simulate rock cave-in. It was concluded that a cave-in would be unlikely. Verification holes confirmed the effectiveness of grouting. Although post grouting micro gravity survey depicted a few anomalies, no anomalies were found to exist by further drilling and excavation.Keywords: grout, stope, rock, properties
Procedia PDF Downloads 3313723 Life Cycle Carbon Dioxide Emissions from the Construction Phase of Highway Sector in China
Authors: Yuanyuan Liu, Yuanqing Wang, Di Li
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Carbon dioxide (CO2) emissions mitigation from road construction activities is one of the potential pathways to deal with climate change due to its higher use of materials, machinery energy consumption, and high quantity of vehicle and equipment fuels for transportation and on-site construction activities. Aiming to assess the environmental impact of the road infrastructure construction activities and to identify hotspots of emissions sources, this study developed a life-cycle CO2 emissions assessment framework covering three stages of material production, to-site and on-site transportation under the guidance of the principle of LCA ISO14040. Then streamlined inventory analysis on sub-processes of each stage was conducted based on the budget files from cases of highway projects in China. The calculation results were normalized into functional unit represented as ton per km per lane. Then a comparison between the amount of emissions from each stage, and sub-process was made to identify the major contributor in the whole highway lifecycle. In addition, the calculating results were used to be compared with results in other countries for understanding the level of CO2 emissions associated with Chinese road infrastructure in the world. The results showed that materials production stage produces the most of the CO2 emissions (for more than 80%), and the production of cement and steel accounts for large quantities of carbon emissions. Life cycle CO2 emissions of fuel and electric energy associated with to-site and on-site transportation vehicle and equipment are a minor component of total life cycle CO2 emissions from highway project construction activities. Bridges and tunnels are dominant large carbon contributor compared to the road segments. The life cycle CO2 emissions of road segment in highway project in China are slightly higher than the estimation results of highways in European countries and USA, about 1500 ton per km per lane. In particularly, the life cycle CO2 emissions of road pavement in majority cities all over the world are about 500 ton per km per lane. However, there is obvious difference between the cities when the estimation on life cycle CO2 emissions of highway projects included bridge and tunnel. The findings of the study could offer decision makers a more comprehensive reference to understand the contribution of road infrastructure to climate change, especially understand the contribution from road infrastructure construction activities in China. In addition, the identified hotspots of emissions sources provide the insights of how to reduce road carbon emissions for development of sustainable transportation.Keywords: carbon dioxide emissions, construction activities, highway, life cycle assessment
Procedia PDF Downloads 2693722 Metformin and Its Combination with Sodium Hydrosulfide Influences Plasma Galectin-3 and CSE/H₂S System in Diabetic Rat's Heart
Authors: I. V. Palamarchuk, N. V. Zaichko
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Background and Aims: Galectin-3 is a marker of subclinical cardiac injury and is elevated in individuals with type 2 diabetes mellitus; while hydrogen sulfide (H₂S), metabolite of sulfur-containing amino acids, is considered having antifibrogenic effects. This study was designed to investigate whether metformin and its combination with NaHS can influence plasma galectin-3 and cystathionine-γ-lyase/hydrogen sulfide (CSE/H₂S) system in diabetic rat’s heart. Methods: 32 healthy male rats (180-250 g) were divided into 4 groups. To induct diabetes, rats (group 2-4) were injected with streptozotocin (STZ, 40 mg/kg/i.p., 0.1 M citrate buffer (pH 4.5). Rats from 3d (STZ+Metf) and 4th (STZ+Metf+NaHS) groups were given metformin (500 mg/kg/day) orally, and rats from 4th (STZ+Metf+NaHS) group were injected sodium hydrosulfide (NaHS, 3 mg/kg/i.p.) once per day starting from 3 to 28 day after streptozotocin injection. Rats of first group (control) were administered the equivalent volumes of 0.9% NaCl. Plasma galectin-3 was measured by ELISA. Rats’ hearts were sampled for determination of H2S by reaction with N,N-Dimethyl-p-phenylenediamine. Determination of CSE gene expression was performed in real time using PCR in the presence of SYBR Green I, using DT-Light detecting amplifier ('DNA-technology', Russia). Results: Induction of streptozotocin diabetes (STZ-diabetes, group 2) was followed by low myocardial H2S concentration and CSE expression (by 35%, p < 0.05 and 60.5%, p < 0.001 respectively, than that in controls), while plasma galectin-3 in this group was significantly higher than in controls (by 3.8 times, p < 0.05). Administration of metformin (group 3) resulted in significantly higher H₂S concentration (by 28.5%, p < 0.05), whereas CSE expression was only by 6% more than that in STZ-diabetes, as well as plasma galectin-3 was only by 14.8% lower in comparison with untreated diabetic rats. The inhibition of H₂S generation and CSE activity by diabetes was greatly attenuated in STZ+Metf+NaHS group. The combination of metformin with NaHS significantly stimulated H₂S production (by 48%, p < 0.05 and 15%, p < 0.05 more than STZ-diabetes and STZ+Metf respectively) and CSE gene expression (by 64.8%, p < 0.05 compared to STZ-diabetes and by 55.4%,p < 0.05 compared to STZ+Metf). Besides, plasma galectin-3 in rats receiving metformin and NaHS was significantly lower by 42%, p < 0.05 and 32.5%, p < 0.05 compared to STZ-diabetes and STZ+Metf groups respectively. Conclusions: To summarize, dysfunction of CSE/H2S system and galectin-3 stimulation was found in streptozotocin-induced diabetic rats. Metformin and its combination with exogenous H2S effectively prevented the development of metabolic changes induced by diabetes. These findings suggest that CSE/H₂S system can be integrated into pathogenesis of diabetic complications through modulation of pro-inflammatory and pro-fibrogenic mediator galectin-3.Keywords: cystathionine-γ-lyase, diabetic heart, galectin-3, hydrogen sulfide, metformin, sodium hydrosulfide
Procedia PDF Downloads 2273721 The Expression of a Novel Gene Encoding an Ankyrin-Repeat Protein, DRA1, Is Regulated by Drought-Responsive Alternative Splicing
Authors: H. Sakamoto, Y. Nakagawara, S. Oguri
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Drought stress is a critical environmental factor that adversely affects crop productivity and quality. Because of their immobile nature, plants have evolved mechanisms to sense and respond to drought stress. We identified a novel locus of Arabidopsis, designated DRA1 (drought responsive ankyrin 1), whose disruption leads to increased drought stress tolerance. DRA1 encodes a transmembrane protein with an ankyrin repeat motif that has been implicated in diverse cellular processes such as signal transduction. RT-PCR analysis revealed that there were at least two splicing variants of DRA1 transcripts in wild type plants. In response to drought stress, the levels of DRA1 transcripts retaining second and third introns were increased, whereas these introns were removed under unstressed conditions. These results suggest that DRA1 protein may negatively regulate plant drought tolerance and that the expression of DRA1 is regulated in response to drought stress by alternative splicing.Keywords: alternative splicing, ankyrin repeat, Arabidopsis, drought tolerance
Procedia PDF Downloads 3263720 Engineering C₃ Plants with SbtA, a Cyanobacterial Transporter, for Enhancing CO₂ Fixation
Authors: Vandana Deopanée Tomar, Gurpreet Kaur Sidhu, Panchsheela Nogia, Rajesh Mehrotra, Sandhya Mehrotra
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The cyanobacterial CO₂ concentrating mechanism (CCM) operates to raise the levels of CO₂ in the vicinity of the main carboxylation enzyme Rubisco which is encapsulated in protein micro compartments called carboxysomes. Thus, due to the presence of CCM, cyanobacterial cells are able to work with high photosynthetic efficiency even at low Ci conditions and can accumulate 1000 folds high internal concentrations of Ci than external environment. Engineering of some useful CCM components into higher plants is one of the plausible approaches to improve their photosynthetic performance. The first step and the simplest approach for attaining this objective would be the transfer of cyanobacterial bicarbonate transporter such as SbtA to inner chloroplast envelope of C₃ plants. For this, SbtA transporter gene from Synechococcus elongatus PCC 7942 was fused to a transit peptide element to generate chimeric constructs in order to direct it to chloroplast inner envelope. Two transit peptides namely, TnaXTP (transit peptide from AT3G56160) and TMDTP (transit peptide from AT2G02590) were shortlisted from Arabidopsis thaliana genome and cloned in plant expression vector pCAMBIA1302 having mgfp5 as a reporter gene. Plant transformation was done by agro infiltration and Agrobacterium mediated co-culture. DNA, RNA, and protein were isolated from the leaves four days post infiltration, and the presence of transgene was confirmed by gene specific PCR (Polymerase Chain Reaction) analysis and by RT-PCR (Reverse Transcription Polymerase Chain Reaction). The expression was confirmed at the protein level by western blotting using anti-GFP primary antibody and horseradish peroxidase (HRP) conjugated secondary antibody. The localization of the protein was detected by confocal microscopy of isolated protoplasts. We observed chloroplastic expression for both the fusion constructs which suggest that the transit peptide sequences are capable of taking the cargo protein to the chloroplasts. These constructs are now being used to generate stable transgenic plants by Agrobacterium mediated transformation. The stability of transgene expression will be analyzed from T₀ to T₂ generation.Keywords: agro infiltration, bicarbonate transporter, carbon concentrating mechanisms, cyanobacteria, SbtA
Procedia PDF Downloads 2213719 A Study on the Conspicuous Consumption, Involvement and Physical and Mental Health of Pet Owners
Authors: Chi-Yueh Hsu, Hsuan-Liang Hsu, Hsiu-Hui Chiang
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This study is to explore the relationship between the conspicuous consumption, leisure involvement and physical and mental health, and to understand the prediction of conspicuous consumption and leisure involvement to physical and mental health. The data was collected and analysed by purposive sampling, and the research objects were the dog walkers in Taiwan area. A total of 300 questionnaires were issued and after shaving the invalid questionnaire, a total of 246 valid samples were collected, and the effective rate was 82%.. The data were analyzed by correlation analysis and multiple stepwise regression analysis. The results showed that there was a significant correlation between conspicuous consumption and leisure involvement, and the conspicuous consumption and leisure involvement of dog walkers have a significant impact on physical and mental health, especially in self-expression, attractiveness and centrality of leisure involvement have a significant impact on physical and mental health.Keywords: walking dog, attractiveness, self-expression, multiple stepwise regression analysis
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