Search results for: cell engineering

Authors: Gulnaz Khan, Meha F. Aftab, Munazza Murtaza, Rizwana S. Waraich

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Advanced glycation end products (AGEs) precursor and its abnormal accumulation cause damage to various tissues and organs. AGEs have pathogenic implication in several diseases including diabetes. Existing AGEs inhibitors are not in clinical use, and there is a need for development of novel inhibitors. The present investigation aimed at identifying the novel AGEs inhibitors and assessing their mechanism of action for treating insulin resistance in mice model of diabetes. Novel derivatives of benzylidene of indan-1,3-dione were synthesized. The compounds were selected to study their action mechanism in improving insulin resistance, in vitro, in human hepatocytes and murine adipocytes and then, in vivo, in mice genetic model of diabetes (db/db). Mice were treated with novel derivatives of benzylidene of indane 1,3-dione. AGEs mediated ROS production was measured by dihydroethidium fluorescence assay. AGEs level in the serum of treated mice was observed by ELISA. Gene expression of receptor for AGEs (RAGE), PPAR-gamma, TNF-alpha and GLUT-4 was evaluated by RT-PCR. Glucose uptake was measured by fluorescent method. Microscopy was used to analyze glycogen synthesis in muscle. Among several derivatives of benzylidene of indan-1,3-dione, IDD-24, demonstrated highest inhibition of AGESs. IDD-24 significantly reduced AGEs formation and expression of receptor for advanced glycation end products (RAGE) in fat, liver of db/db mice. Suppression of AGEs mediated ROS production was also observed in hepatocytes and fat cell, after treatment with IDD-24. Glycogen synthesis was increased in muscle tissue of mice treated with IDD-24. In adipocytes, IDD-24 prevented AGEs induced reduced glucose uptake. Mice treated with IDD-24 exhibited increased glucose tolerance, serum adiponectin levels and decreased insulin resistance. The result of present study suggested that IDD-24 can be a possible treatment target to address glycotoxins induced insulin resistance.

Keywords: advance glycation end product, hyperglycemia, indan-1, 3-dione, insulin resistance

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Authors: Atlal El-Assaad

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Viruses change with time through mutations and result in new variants that may persist or disappear. A Mutation refers to an actual change in the virus genetic sequence, and a variant is a viral genome that may contain one or more mutations. Critical mutations may cause the virus to be more transmissible, with high disease severity, and more vulnerable to diagnostics, therapeutics, and vaccines. Thus, variants carrying such mutations may increase the risk to human health and are considered variants of concern (VOC). Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) - the contagious in humans, positive-sense single-stranded RNA virus that caused coronavirus disease 2019 (COVID-19) - has been studied thoroughly, and several variants were revealed across the world with their corresponding mutations. SARS-CoV-2 has four structural proteins, known as the S (spike), E (envelope), M (membrane), and N (nucleocapsid) proteins, but prior study and vaccines development focused on genetic mutations in the S protein due to its vital role in allowing the virus to attach and fuse with the membrane of a host cell. Specifically, subunit S1 catalyzes attachment, whereas subunit S2 mediates fusion. In this perspective, we studied all charged amino acid mutations of the SARS-CoV-2 viral spike protein S1 when bound to Antibody CC12.1 in a crystal structure and assessed the effect of different mutations. We generated all missense mutants of SARS-CoV-2 protein amino acids (AAs) within the SARS-CoV-2:CC12.1 complex model. To generate the family of mutants in each complex, we mutated every charged amino acid with all other charged amino acids (Lysine (K), Arginine (R), Glutamic Acid (E), and Aspartic Acid (D)) and studied the new binding of the complex after each mutation. We applied Poisson-Boltzmann electrostatic calculations feeding into free energy calculations to determine the effect of each mutation on binding. After analyzing our data, we identified charged amino acids keys for binding. Furthermore, we validated those findings against published experimental genetic data. Our results are the first to propose in silico potential life-threatening mutations of SARS-CoV-2 beyond the present mutations found in the five common variants found worldwide.

Keywords: SARS-CoV-2, variant, ionic amino acid, protein-protein interactions, missense mutation, AESOP

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Authors: Zhidong Zhang, Yingchen Yang

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In this study, a diagnostic assessment model for Mastery Engineering Learning was established based on a group of undergraduate students who studied in an engineering course. A diagnostic assessment model can examine both students' learning process and report achievement results. One very unique characteristic is that the diagnostic assessment model can recognize the errors and anything blocking students in their learning processes. The feedback is provided to help students to know how to solve the learning problems with alternative strategies and help the instructor to find alternative pedagogical strategies in the instructional designs. Dynamics is a core course in which is a common course being shared by several engineering programs. This course is a very challenging for engineering students to solve the problems. Thus knowledge acquisition and problem-solving skills are crucial for student success. Therefore, developing an effective and valid assessment model for student learning are of great importance. Diagnostic assessment is such a model which can provide effective feedback for both students and instructor in the mastery of engineering learning.

Keywords: diagnostic assessment, mastery learning, engineering, bayesian network model, learning processes

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Authors: Jamboor K. Vishwanatha

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Among the potent anti-cancer agents, curcumin has been found to be very efficacious against various cancer cells. Despite multiple medicinal benefits of curcumin, poor water solubility, poor physiochemical properties and low bioavailability continue to pose major challenges in developing a formulation for clinical efficacy. To improve its potential application in the clinical area, we formulated poly lactic-co-glycolic acid (PLGA) nanoparticles. The PLGA nanoparticles were formulated using solid-oil/water emulsion solvent evaporation method and then characterized for percent yield, encapsulation efficiency, surface morphology, particle size, drug distribution within nanoparticles and drug polymer interaction. Our studies showed the successful formation of smooth and spherical curcumin loaded PLGA nanoparticles with a high percent yield of about 92.01±0.13% and an encapsulation efficiency of 90.88±0.14%. The mean particle size of the nanoparticles was found to be 145nm. The in vitro drug release profile showed 55-60% drug release from the nanoparticles over a period of 24 hours with continued sustained release over a period of 8 days. Exposure to curcumin loaded nanoparticles resulted in reduced cell viability of cancer cells compared to normal cells. We used a novel non-covalent insertion of a homo-bifunctional spacer for targeted delivery of curcumin to various cancer cells. Functionalized nanoparticles for antibody/targeting agent conjugation was prepared using a cross-linking ligand, bis(sulfosuccinimidyl) suberate (BS3), which has reactive carboxyl group to conjugate efficiently to the primary amino groups of the targeting agents. In our studies, we demonstrated successful conjugation of antibodies, Annexin A2 or prostate specific membrane antigen (PSMA), to curcumin loaded PLGA nanoparticles for targeting to prostate and breast cancer cells. The percent antibody attachment to PLGA nanoparticles was found to be 92.8%. Efficient intra-cellular uptake of the targeted nanoparticles was observed in the cancer cells. These results have emphasized the potential of our multifunctional curcumin nanoparticles to improve the clinical efficacy of curcumin therapy in patients with cancer.

Keywords: polymeric nanoparticles, cancer therapy, sustained release, curcumin

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Authors: Natalia Moreno-Castellanos, Amaia Rodriguez, Gema Frühbeck

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Introduction: In adipocytes, the cytoskeleton undergoes important expression and distribution in adipocytes rearrangements during adipogenesis and in obesity. Indeed, a role for these proteins in the regulation of adipocyte differentiation and response to insulin has been demonstrated. Recently, septins have been considered as new components of the cytoskeletal network that interact with other cytoskeletal elements (actin and tubulin) profoundly modifying their dynamics. However, these proteins have not been characterized as yet in adipose tissue. In this work, were examined the cellular, molecular and functional features of a member of this family, septin 11 (SEPT11), in adipocytes and evaluated the impact of obesity on the expression of this protein in human adipose tissue. Methods: Adipose gene and protein expression levels of SEPT11 were analysed in human samples. SEPT11 distribution was evaluated by immunocytochemistry, electronic microscopy, and subcellular fractionation techniques. GST-pull down, immunoprecipitation and a Yeast-Two Hybrid (Y2H) screening were used to identify the SEPT11 interactome. Gene silencing was employed to assess the role of SEPT11 in the regulation of insulin signaling and lipid metabolism in adipocytes. Results: SEPT11 is expressed in human adipocytes, and its levels increased in both omental and subcutaneous adipose tissue in obesity, with SEPT11 mRNA content positively correlating with parameters of insulin resistance in subcutaneous fat. In non-stimulated adipocytes, SEPT11 immunoreactivity showed a ring-like distribution at the cell surface and associated to caveolae. Biochemical analyses showed that SEPT11 interacted with the main component of caveolae, caveolin-1 (CAV1) as well as with the fatty acid-binding protein, FABP5. Notably, the three proteins redistributed and co-localized at the surface of lipid droplets upon exposure of adipocytes to oleate. In this line, SEPT11 silencing in 3T3-L1 adipocytes impaired insulin signaling and decreased insulin-induced lipogenesis. Conclusions: Those findings demonstrate that SEPT11 is a novel component of the adipocyte cytoskeleton that plays an important role in the regulation of lipid traffic, metabolism and can thus represent a potential biomarker of insulin resistance in obesity in adipocytes through its interaction with both CAV1 and FABP5.

Keywords: caveolae, lipid metabolism, obesity, septins

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Authors: Yosep Chong, Yejin Kim, Jingyun Choi, Hwanjo Yu, Eun Jung Lee, Chang Suk Kang

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For the past decades, immunohistochemistry (IHC) has been playing an important role in the diagnosis of human neoplasms, by helping pathologists to make a clearer decision on differential diagnosis, subtyping, personalized treatment plan, and finally prognosis prediction. However, the IHC performed in various tumors of daily practice often shows conflicting and very challenging results to interpret. Even comprehensive diagnosis synthesizing clinical, histologic and immunohistochemical findings can be helpless in some twisted cases. Another important issue is that the IHC data is increasing exponentially and more and more information have to be taken into account. For this reason, we reached an idea to develop an expert supporting system to help pathologists to make a better decision in diagnosing human neoplasms with IHC results. We gave probabilistic decision tree algorithm and tested the algorithm with real case data of lymphoid neoplasms, in which the IHC profile is more important to make a proper diagnosis than other human neoplasms. We designed probabilistic decision tree based on Bayesian theorem, program computational process using MATLAB (The MathWorks, Inc., USA) and prepared IHC profile database (about 104 disease category and 88 IHC antibodies) based on WHO classification by reviewing the literature. The initial probability of each neoplasm was set with the epidemiologic data of lymphoid neoplasm in Korea. With the IHC results of 131 patients sequentially selected, top three presumptive diagnoses for each case were made and compared with the original diagnoses. After the review of the data, 124 out of 131 were used for final analysis. As a result, the presumptive diagnoses were concordant with the original diagnoses in 118 cases (93.7%). The major reason of discordant cases was that the similarity of the IHC profile between two or three different neoplasms. The expert supporting system algorithm presented in this study is in its elementary stage and need more optimization using more advanced technology such as deep-learning with data of real cases, especially in differentiating T-cell lymphomas. Although it needs more refinement, it may be used to aid pathological decision making in future. A further application to determine IHC antibodies for a certain subset of differential diagnoses might be possible in near future.

Keywords: database, expert supporting system, immunohistochemistry, probabilistic decision tree

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Authors: D. S. Zhuravkova, A. G. Temnikov, O. S. Belova, L. L. Chernensky, T. K. Gerastenok, I. Y. Kalugina, N. Y. Lysov, A.V. Orlov

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Investigation of the discharges from the artificial charged water aerosol clouds in presence of the arrays of the model hydrometeors could help to receive the new data about the peculiarities of the return stroke formation between the thundercloud and the ground when the large volumes of the hail particles participate in the lightning discharge initiation and propagation stimulation. Artificial charged water aerosol clouds of the negative or positive polarity with the potential up to one million volts have been used. Hail has been simulated by the group of the conductive model hydrometeors of the different form. Parameters of the impulse current of the main stage of the discharge between the artificial positively and negatively charged water aerosol clouds and the ground in presence of the model hydrometeors array and of its corresponding electromagnetic radiation have been determined. It was established that the parameters of the array of the model hydrometeors influence on the parameters of the main stage of the discharge between the artificial thundercloud cell and the ground. The maximal values of the main stage current impulse parameters and the electromagnetic radiation registered by the plate antennas have been found for the array of the model hydrometeors of the cylinder revolution form for the negatively charged aerosol cloud and for the array of the hydrometeors of the plate rhombus form for the positively charged aerosol cloud, correspondingly. It was found that parameters of the main stage of the discharge between the artificial charged water aerosol cloud and the ground in presence of the model hydrometeor array of the different considered forms depend on the polarity of the artificial charged aerosol cloud. In average, for all forms of the investigated model hydrometeors arrays, the values of the amplitude and the current rise of the main stage impulse current and the amplitude of the corresponding electromagnetic radiation for the artificial charged aerosol cloud of the positive polarity were in 1.1-1.9 times higher than for the charged aerosol cloud of the negative polarity. Thus, the received results could indicate to the possible more important role of the big volumes of the large hail arrays in the thundercloud on the parameters of the return stroke for the positive lightning.

Keywords: main stage of discharge, hydrometeor form, lightning parameters, negative and positive artificial charged aerosol cloud

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Authors: Ahmed Abdulrahman, Jalal Foroozesh

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CO₂ based enhanced oil recovery (EOR) techniques have gained massive attention from major oil firms since they resolve the industry's two main concerns of CO₂ contribution to the greenhouse effect and the declined oil production. Carbonated water injection (CWI) is a promising EOR technique that promotes safe and economic CO₂ storage; moreover, it mitigates the pitfalls of CO₂ injection, which include low sweep efficiency, early CO₂ breakthrough, and the risk of CO₂ leakage in fractured formations. One of the main challenges that hinder the wide adoption of this EOR technique is the complexity of accurate modeling of the kinetics of CO₂ mass transfer. The mechanisms of CO₂ mass transfer during CWI include the slow and gradual cross-phase CO₂ diffusion from carbonated water (CW) to the oil phase and the CO₂ dispersion (within phase diffusion and mechanical mixing), which affects the oil physical properties and the spatial spreading of CO₂ inside the reservoir. A 2D non-equilibrium compositional simulator has been developed using a fully implicit finite difference approximation. The material balance term (k) was added to the governing equation to account for the slow cross-phase diffusion of CO₂ from CW to the oil within the gird cell. Also, longitudinal and transverse dispersion coefficients have been added to account for CO₂ spatial distribution inside the oil phase. The CO₂-oil diffusion coefficient was calculated using the Sigmund correlation, while a scale-dependent dispersivity was used to calculate CO₂ mechanical mixing. It was found that the CO₂-oil diffusion mechanism has a minor impact on oil recovery, but it tends to increase the amount of CO₂ stored inside the formation and slightly alters the residual oil properties. On the other hand, the mechanical mixing mechanism has a huge impact on CO₂ spatial spreading (accurate prediction of CO₂ production) and the noticeable change in oil physical properties tends to increase the recovery factor. A sensitivity analysis has been done to investigate the effect of formation heterogeneity (porosity, permeability) and injection rate, it was found that the formation heterogeneity tends to increase CO₂ dispersion coefficients, and a low injection rate should be implemented during CWI.

Keywords: CO₂ mass transfer, carbonated water injection, CO₂ dispersion, CO₂ diffusion, cross phase CO₂ diffusion, within phase CO2 diffusion, CO₂ mechanical mixing, non-equilibrium simulation

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Authors: Ifeoma C. Afiaenyi, Elizabeth K. Ngwu, Rufina N. B. Ayogu

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Diabetes has crept into the rural areas of Nigeria, causing devastating effects on its sufferers; most of them could not afford diabetic medications. Moringa oleifera has been used extensively in animal models to demonstrate its antilipidaemic and haematinic qualities; however, there is a scarcity of data on the effect of graded levels of Moringa oleifera leaves on the lipid profile and hematological parameters in human diabetic subjects. The study determined the effect of Moringa oleifera leaves on the lipid profile and hematological parameters of type 2 diabetic subjects in Ukehe, a rural Nigerian community. Twenty-four adult male and female diabetic subjects were purposively selected for the study. These subjects were shared into four groups of six subjects each. The diets used in the study were isocaloric. A control group (diabetics, group 1) was fed diets without Moringa oleifera leaves. Experimental groups 2, 3 and 4 received 20g, 40g and 60g of Moringa oleifera leaves daily, respectively, in addition to the diets. The subjects' lipid profile and hematological parameters were measured prior to the feeding trial and at the end of the feeding trial. The feeding trial lasted for fourteen days. The data obtained were analyzed using the computer program Statistical Product for Service Solution (SPSS) for windows version 21. A Paired-samples t-test was used to compare the means of values collected before and after the feeding trial within the groups and significance was accepted at p < 0.05. There was a non-significant (p > 0.05) decrease in the mean total cholesterol of the subjects in groups 1, 2 and 3 after the feeding trial. There was a non-significant (p > 0.05) decrease in the mean triglyceride levels of the subjects in group 1 after the feeding trial. Groups 1 and 3 subjects had a non-significant (p > 0.05) decrease in their mean low-density lipoprotein (LDL) cholesterol after the feeding trial. Groups 1, 2 and 4 had a significant (p < 0.05) increase in their mean high-density lipoprotein (HDL) cholesterol after the feeding trial. A significant (p < 0.05) decrease in the mean hemoglobin level was observed only in group 4 subjects. Similarly, there was a significant (p < 0.05) decrease in the mean packed cell volume of group 4 subjects. It was only in group 4 that a significant (p < 0.05) decrease in the mean white blood cells of the subjects was also observed. The changes observed in the parameters assessed were not dose-dependent. Therefore, a similar study of longer duration and more samples is imperative to authenticate these results.

Keywords: anemia, diabetic subjects, lipid profile, moringa oleifera

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Authors: Sajewicz-Krukowska Joanna, Domańska-Blicharz Katarzyna, Tarasiuk Karolina, Marzec-Kotarska Barbara

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Astroviral infections pose a significant problem in the poultry industry, leading to multiple adverse effects such as decreased egg production, breeding disorders, poor weight gain, and even increased mortality. Commonly observed chicken astrovirus (CAstV) was recently reported to be responsible for "white chicks syndrome" associated with increased embryo/chick mortality. The CAstV-mediated pathogenesis in chicken occurs due to complex interactions between the infectious pathogen and the immune system. Many aspects of CAstV-chicken interactions remain unclear, and there is no information available regarding gene expression changes in the chicken's spleen in response to CAstV infection. We aimed to investigate the molecular background triggered by CAstV infection. Ten 21-day-old SPF White Leghorn chickens were divided into two groups of 5 birds each. One group was inoculated with CAstV, and the other was used as the negative control. On 4th dpi, spleen samples were collected and immediately frozen at -70°C for RNA isolation. We analysed transcriptional profiles of the chickens' spleens at the 4th day following infection using RNA-seq to establish differentially expressed genes (DEGs). The RNA-seq findings were verified by quantitative real-time PCR (qRT-PCR). A total of 31959 transcripts were identified in response to CAstV infection. Eventually 45 DEGs (p-value<0.05; Log2Foldchange>1)were recognized in the spleen after CAstV infection (26 upregulated DEGs and 19 downregulated DEGs). qRT-PCR performed on 4 genes (IFIT5, OASL, RASD1, DDX60) confirmed RNAseq results. Top differentially expressed genes belonged to novel putative IFN-induced CAstV restriction factors. Most of the DEGs were associated with RIG-I–like signalling pathway or, more generally, with an innate antiviral response(upregulated: BLEC3, CMPK2, IFIT5, OASL, DDX60, IFI6, and downregulated: SPIK5, SELENOP, HSPA2, TMEM158, RASD1, YWHAB). The study provided a global analysis of host transcriptional changes that occur during CAstV infection in vivo and proved the cell cycle in the spleen and immune signalling in chickens were predominantly affected upon CAstV infection.

Keywords: chicken astrovirus, CastV, RNA-seq, transcriptome, spleen

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Authors: Mahbuba Rahman, Sabri Boughorbel, Scott Presnell, Charlie Quinn, Darawan Rinchai, Damien Chaussabel, Nico Marr

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Collections of large-scale datasets made available in public repositories can be used to identify and fill gaps in biomedical knowledge. But first, these data need to be made readily accessible to researchers for analysis and interpretation. Here a collection of transcriptome datasets was made available to investigate the functional programming of human hematopoietic cells in early life. Thirty two datasets were retrieved from the NCBI Gene Expression Omnibus (GEO) and loaded in a custom, interactive web application called the Gene Expression browser (GXB), designed for visualization and query of integrated large-scale data. Multiple sample groupings and gene rank lists were created based on the study design and variables in each dataset. Web links to customized graphical views can be generated by users and subsequently be used to graphically present data in manuscripts for publication. The GXB tool also enables browsing of a single gene across datasets, which can provide information on the role of a given molecule across biological systems. The dataset collection is available online. As a proof-of-principle, one of the datasets (GSE25087) was re-analyzed to identify genes that are differentially expressed by regulatory T cells in early life. Re-analysis of this dataset and a cross-study comparison using multiple other datasets in the above mentioned collection revealed that PMCH, a gene encoding a precursor of melanin-concentrating hormone (MCH), a cyclic neuropeptide, is highly expressed in a variety of other hematopoietic cell types, including neonatal erythroid cells as well as plasmacytoid dendritic cells upon viral infection. Our findings suggest an as yet unrecognized role of MCH in immune regulation, thereby highlighting the unique potential of the curated dataset collection and systems biology approach to generate new hypotheses which can be tested in future mechanistic studies.

Keywords: early-life, GEO datasets, PMCH, interactive query, systems biology

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Authors: Adnan A. Kadi, Nasser S. Al-Shakliah, Haitham Al-Rabiah

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Zorifertinib is a novel, potent, oral, a small molecule used to treat non-small cell lung cancer (NSCLC). zorifertinib is an Epidermal Growth Factor Receptor (EGFR) inhibitor and has good blood–brain barrier permeability for (NSCLC) patients with EGFR mutations. zorifertinibis currently at phase II/III clinical trials. The current research reports the characterization and identification of in vitro, in vivo and reactive intermediates of zorifertinib. Prediction of susceptible sites of metabolism and reactivity pathways (cyanide and GSH) of zorifertinib were performed by the Xenosite web predictor tool. In-vitro metabolites of zorifertinib were performed by incubation with rat liver microsomes (RLMs) and isolated perfused rat liver hepatocytes. Extraction of zorifertinib and it's in vitro metabolites from the incubation mixtures were done by protein precipitation. In vivo metabolism was done by giving a single oral dose of zorifertinib(10 mg/Kg) to Sprague Dawely rats in metabolic cages by using oral gavage. Urine was gathered and filtered at specific time intervals (0, 6, 12, 18, 24, 48, 72,96and 120 hr) from zorifertinib dosing. A similar volume of ACN was added to each collected urine sample. Both layers (organic and aqueous) were injected into liquid chromatography ion trap mass spectrometry(LC-IT-MS) to detect vivozorifertinib metabolites. N-methyl piperizine ring and quinazoline group of zorifertinib undergoe metabolism forming iminium and electro deficient conjugated system respectively, which are very reactive toward nucleophilic macromolecules. Incubation of zorifertinib with RLMs in the presence of 1.0 mM KCN and 1.0 Mm glutathione were made to check reactive metabolites as it is often responsible for toxicities associated with this drug. For in vitro metabolites there were nine in vitro phase I metabolites, four in vitro phase II metabolites, eleven reactive metabolites(three cyano adducts, five GSH conjugates metabolites, and three methoxy metabolites of zorifertinib were detected by LC-IT-MS. For in vivo metabolites, there were eight in vivo phase I, tenin vivo phase II metabolitesofzorifertinib were detected by LC-IT-MS. In vitro and in vivo phase I metabolic pathways wereN- demthylation, O-demethylation, hydroxylation, reduction, defluorination, and dechlorination. In vivo phase II metabolic reaction was direct conjugation of zorifertinib with glucuronic acid and sulphate.

Keywords: in vivo metabolites, in vitro metabolites, cyano adducts, GSH conjugate

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Authors: Aleya Ferdausi, Meriel Jones, Anthony Halls

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The Amaryllidaceae genus Narcissus contains secondary metabolites, which are important sources of bioactive compounds such as pharmaceuticals indicating that their biological activity extends from the native plant to humans. Transcriptome analysis (RNA-seq) is an effective platform for the identification and functional characterization of candidate genes as well as to identify genes encoding uncharacterized enzymes. The biotechnological production of secondary metabolites in plant cell or organ cultures has become a tempting alternative to the extraction of whole plant material. The biochemical pathways for the production of secondary metabolites require primary metabolites to undergo a series of modifications catalyzed by enzymes such as cytochrome P450s, methyltransferases, glycosyltransferases, and acyltransferases. Differential gene expression analysis of Narcissus was obtained from two conditions, i.e. field and in vitro callus. Callus was obtained from modified MS (Murashige and Skoog) media supplemented with growth regulators and twin-scale explants from Narcissus cv. Carlton bulb. A total of 2153 differentially expressed transcripts were detected in Narcissus bulb and in vitro callus, and 78.95% of those were annotated. It showed the expression of genes involved in the biosynthesis of alkaloids were present in both conditions i.e. cytochrome P450s, O-methyltransferase (OMTs), NADP/NADPH dehydrogenases or reductases, SAM-synthetases or decarboxylases, 3-ketoacyl-CoA, acyl-CoA, cinnamoyl-CoA, cinnamate 4-hydroxylase, alcohol dehydrogenase, caffeic acid, N-methyltransferase, and NADPH-cytochrome P450s. However, cytochrome P450s and OMTs involved in the later stage of Amaryllidaceae alkaloids biosynthesis were mainly up-regulated in field samples. Whereas, the enzymes involved in initial biosynthetic pathways i.e. fructose biphosphate adolase, aminotransferases, dehydrogenases, hydroxyl methyl glutarate and glutamate synthase leading to the biosynthesis of precursors; tyrosine, phenylalanine and tryptophan for secondary metabolites were up-regulated in callus. The knowledge of probable genes involved in secondary metabolism and their regulation in different tissues will provide insight into the Narcissus plant biology related to alkaloid production.

Keywords: narcissus, callus, transcriptomics, secondary metabolites

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Authors: Asma Chbani, Douaa Salim, Josephine Al Alam, Pascale De Caro

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Penicillium digitatum, the causal agent of citrus green mold, is responsible for 90% of post-harvest losses. Chemical fungicides remain the most used products for protection against this pathogen but are also responsible for damage to human health and the environment. The aim of this study is to evaluate the ability of Spinacia oleracea extract to serve as biological control agents, an alternative to harmful synthetic fungicides, against orange decay for storing fruit caused by P. digitatum. In this study, we studied the implication of a crude extract of a green plant, Spinacia oleracea, in the protection of oranges against P. digitatum. Thus, in vivo antifungal tests as well as adhesion test were done. For in vivo antifungal test, oranges were pulverized with the prepared crude extracts at different concentrations ranged from 25 g L⁻¹ to 200 g L⁻¹, contaminated by the fungus and then observed during 8 weeks for their macroscopic changes at 24°C. For adhesion test, the adhesion index is defined as the number of Penicillium digitatum spores fixed per orange cell. An index greater than 25 is the indicator of a strong adhesion, whereas for an index less than 10, the adhesion is low. Ten orange cells were examined in triplicate for each extract, and the averages of adherent cells were calculated. Obtained results showed an inhibitory activity of the Penicillium development with the aqueous extract of dry Spinacia oleracea with a concentration of 50 g L⁻¹ considered as the minimal protective concentration. The prepared extracts showed a greater inhibition of the development of P. digitatum up to 10 weeks, even greater than the fungicide control Nystatin. Adhesion test’s results showed that the adhesion of P. digitatum spores to the epidermal cells of oranges in the presence of the crude spinach leaves extract is weak; the mean of the obtained adhesion index was estimated to 2.7. However, a high adhesion was observed with water used a negative control. In conclusion, all these results confirm that the use of this green plant highly rich in chlorophyll having several phytotherapeutic activities, could be employed as a great treatment for protection of oranges against mold and also as an alternative for chemical fungicides.

Keywords: Penicillium digitatum, Spinacia oleracea, oranges, biological control, postharvest diseases

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Authors: Mahfoud Djebbar

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The past two decades or so have witnessed phenomenal growth of the Islamic financial services industry. The whole industry has been thriving at about 15 percent per annum. This development entails the Islamic financial engineering, IFE, to some kind of crossroads, lagging behind its conventional counterpart. Therefore, IFE, and particularly traded products development, and in order to achieve its goals, two approaches are available, i.e., replicating engineering and innovative engineering. We also try to emphasis the innovative strategy since it guards the Islamic identity of different financial products and processes, and thereby, improves the creativity in the Islamic financial industry. The attempt also centers on sukukization (Islamic securitization), innovation, liquidity management, and risk management and hedging in the Islamic financial system. Finally, the challenges facing IFE are also addressed.

Keywords: islamic financial engineering, hedging and risk management, innovation, securitization, money market instruments, islamic capital markets

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Authors: Jianming Cai

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Exposure to ionizing radiation causes severe damage to human body and an safe and effective radioprotector is urgently required for alleviating radiation damage. In 2008, flagellin, an agonist of TLR5, was found to exert radioprotective effects on radiation injury through activating NF-kB signaling pathway. From then, the radioprotective effects of TLR ligands has shed new lights on radiation protection. CpG-ODN is an unmethylated oligonucleotide which activates TLR9 signaling pathway. In this study, we demonstrated that CpG-ODN has therapeutic effects on radiation injuries induced by γ ray and 12C6+ heavy ion particles. Our data showed that CpG-ODN increased the survival rate of mice after whole body irradiation and increased the number of leukocytes as well as the bone marrow cells. CpG-ODN also alleviated radiation damage on intestinal crypt through regulating apoptosis signaling pathway including bcl2, bax, and caspase 3 etc. By using a radiation-induced pulmonary fibrosis model, we found that CpG-ODN could alleviate structural damage, within 20 week after whole–thorax 15Gy irradiation. In this model, Th1/Th2 imbalance induced by irradiation was also reversed by CpG-ODN. We also found that TGFβ-Smad signaling pathway was regulated by CpG-ODN, which accounts for the therapeutic effects of CpG-ODN in radiation-induced pulmonary injury. On another hand, for high LET radiation protection, we investigated protective effects of CpG-ODN against 12C6+ heavy ion irradiation and found that after CpG-ODN treatment, the apoptosis and cell cycle arrest induced by 12C6+ irradiation was reduced. CpG-ODN also reduced the expression of Bax and caspase 3, while increased the level of bcl2. Then we detected the effect of CpG-ODN on heavy ion induced immune dysfunction. Our data showed that CpG-ODN increased the survival rate of mice and also the leukocytes after 12C6+ irradiation. Besides, the structural damage of immune organ such as thymus and spleen was also alleviated by CpG-ODN treatment. In conclusion, we found that TLR9 ligand, CpG-ODN reduced radiation injuries in response to γ ray and 12C6+ heavy ion irradiation. On one hand, CpG-ODN inhibited the activation of apoptosis induced by radiation through regulating bcl2, bax and caspase 3. On another hand, through activating TLR9, CpG-ODN recruit MyD88-IRAK-TRAF6 complex, activating TAK1, IRF5 and NF-kB pathway, and thus alleviates radiation damage. This study provides novel insights into protection and therapy of radiation damages.

Keywords: TLR9, CpG-ODN, radiation injury, high LET radiation

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Authors: Sinethemba Yakobi, Lindiwe Zuma, Ofentse Pooe

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Gonococcal infections present a notable public health issue, and the major approach for treatment involves using β-lactam antibiotics that specifically target penicillin-binding protein 2 (PBP2) in Neisseria gonorrhoeae. This study examines the influence of flavonoids, namely rutin, on the structural changes of PBP2 in both penicillin-resistant (FA6140) and penicillin-susceptible (FA19) strains. The research clarifies the structural effects of particular mutations, such as inserting an aspartate residue at position 345 (Asp-345a) in the PBP2 protein. The strain FA6140, which is resistant to penicillin, shows specific changes that lead to a decrease in penicillin binding. These mutations, namely P551S and F504L, significantly impact the pace at which acylation occurs and the stability of the strain under high temperatures. Molecular docking analyses investigate the antibacterial activities of rutin and other phytocompounds, emphasizing its exceptional binding affinity and potential as an inhibitor of PBP2. Quercetin and protocatechuic acid have encouraging antibacterial effectiveness, with quercetin displaying characteristics similar to those of drugs. Molecular dynamics simulations offer a detailed comprehension of the interactions between flavonoids and PBP2, highlighting rutin's exceptional antioxidant effects and strong affinity for the substrate binding site. The study's wider ramifications pertain to the pressing requirement for antiviral treatments in the context of the ongoing COVID-19 epidemic. Flavonoids have a strong affinity for binding to PBP2, indicating their potential as inhibitors to impair cell wall formation in N. gonorrhoeae. Ultimately, this study provides extensive knowledge on the interactions between proteins and ligands, the dynamics of the structure, and the ability of flavonoids to combat penicillin-resistant N. gonorrhoeae bacteria. The verified simulation outcomes establish a basis for creating potent inhibitors and medicinal therapies to combat infectious illnesses.

Keywords: phytochemicals, penicillin-binding protein 2, gonococcal infection, ligand-protein interaction, binding energy, neisseria gonorrhoeae FA19, neisseria gonorrhoeae FA6140, flavonoids

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Authors: Tajinder Pal Singh

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Teaching of mathematics to engineering students is an open ended problem in education. The main goal of mathematics learning for engineering students is the ability of applying a wide range of mathematical techniques and skills in their engineering classes and later in their professional work. Most of the undergraduate engineering students and faculties feels that no efforts and attempts are made to demonstrate the applicability of various topics of mathematics that are taught thus making mathematics unavoidable for some engineering faculty and their students. The lack of understanding of concepts in engineering mathematics may hinder the understanding of other concepts or even subjects. However, for most undergraduate engineering students, mathematics is one of the most difficult courses in their field of study. Most of the engineering students never understood mathematics or they never liked it because it was too abstract for them and they could never relate to it. A right balance of application and concept based teaching can only fulfill the objectives of teaching mathematics to engineering students. It will surely improve and enhance their problem solving and creative thinking skills. In this paper, some practical (informal) ways of making mathematics-teaching application based for the engineering students is discussed. An attempt is made to understand the present state of teaching mathematics in engineering colleges. The weaknesses and strengths of the current teaching approach are elaborated. Some of the causes of unpopularity of mathematics subject are analyzed and a few pragmatic suggestions have been made. Faculty in mathematics courses should spend more time discussing the applications as well as the conceptual underpinnings rather than focus solely on strategies and techniques to solve problems. They should also introduce more ‘word’ problems as these problems are commonly encountered in engineering courses. Overspecialization in engineering education should not occur at the expense of (or by diluting) mathematics and basic sciences. The role of engineering education is to provide the fundamental (basic) knowledge and to teach the students simple methodology of self-learning and self-development. All these issues would be better addressed if mathematics and engineering faculty join hands together to plan and design the learning experiences for the students who take their classes. When faculties stop competing against each other and start competing against the situation, they will perform better. Without creating any administrative hassles these suggestions can be used by any young inexperienced faculty of mathematics to inspire engineering students to learn engineering mathematics effectively.

Keywords: application based learning, conceptual learning, engineering mathematics, word problem

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Authors: Malay Niraj, Praveen Kumar

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The present study is a new approach for improving equipment life and Overall Equipment Efficiency (O.E.E.) through suitable maintenance practice with the help of value engineering. Value engineering is a one of the most powerful decision-making techniques which depend on many factors. The improvements are the result of recommendations made by multidisciplinary teams representing all parties involved. VE is a rigorous, systematic effort to improve the OEE and optimize the life cycle cost of a facility. The study describes problems in maintenance arising due to the absence of having clear criteria and strong decision constrain how to maintain failing equipment. Using factor comparisons, the study has been made between different maintenance practices and finally best maintenance practice based on value engineering technique has been selected.

Keywords: maintenance strategy, overall equipment efficiency, value engineering, decision-making

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Authors: Sireetorn Kuharat, Anwar Beg

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In this study, multi-mode heat transfer characteristics of spacecraft solar collectors are investigated computationally. Two-dimensional steady-state incompressible laminar Newtonian viscous convection-radiative heat transfer in a rectangular solar collector geometry. The ANSYS FLUENT finite volume code (version 17.2) is employed to simulate the thermo-fluid characteristics. Several radiative transfer models are employed which are available in the ANSYS workbench, including the classical Rosseland flux model and the more elegant P1 flux model. Mesh-independence tests are conducted. Validation of the simulations is conducted with a computational Harlow-Welch MAC (Marker and Cell) finite difference method and excellent correlation. The influence of aspect ratio, Prandtl number (Pr), Rayleigh number (Ra) and radiative flux model on temperature, isotherms, velocity, the pressure is evaluated and visualized in color plots. Additionally, the local convective heat flux is computed and solutions are compared with the MAC solver for various buoyancy effects (e.g. Ra = 10,000,000) achieving excellent agreement. The P1 model is shown to better predict the actual influence of solar radiative flux on thermal fluid behavior compared with the limited Rosseland model. With increasing Rayleigh numbers the hot zone emanating from the base of the collector is found to penetrate deeper into the collector and rises symmetrically dividing into two vortex regions with very high buoyancy effect (Ra >100,000). With increasing Prandtl number (three gas cases are examined respectively hydrogen gas mixture, air and ammonia gas) there is also a progressive incursion of the hot zone at the solar collector base higher into the solar collector space and simultaneously a greater asymmetric behavior of the dual isothermal zones. With increasing aspect ratio (wider base relative to the height of the solar collector geometry) there is a greater thermal convection pattern around the whole geometry, higher temperatures and the elimination of the cold upper zone associated with lower aspect ratio.

Keywords: thermal convection, radiative heat transfer, solar collector, Rayleigh number

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Authors: Mahdi Eskandarlou, Mehrdad Taghipour

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Background: Partial-thickness skin graft is the cornerstone for scalp defect repair. Routine donor sites include abdomen, thighs, and buttocks. Given the potential side effects following harvesting from these sites and the potential advantages of harvesting from scalp (broad surface, rapid healing, and better cosmetics results), this study is trying to compare the outcomes of graft harvesting from scalp and lower limb. Methods: This clinical trial is conducted among a sample number of 40 partial thickness graft candidates (20 case and 20 control group) with scalp defect presenting to plastic surgery clinic at Besat Hospital during the time period between 2018 and 2019. Sampling was done by simple randomization using random digit table. Data gathering was performed using a designated checklist. The donor site in case group and control group was scalp and lower limb, respectively. The resultant data were analyzed using chi-squared and t-test and SPPS version 21 (SPSS Statistics for Windows, Version 21.0. Armonk, NY: IBM Corp). Results: Of the total 40 patients participating in this study, 28 patients (70%) were male, and 12 (30%) were female with and mean age of 63.62 ± 09.73 years. Hypertension and diabetes mellitus were the most common comorbidities among patients with basal cell carcinoma (BCC) and trauma being the most common etiology for the defects. There was a statistically meaningful relationship between two groups regarding the etiology of defect (P=0.02). The most common anatomic location of defect for case and control groups was temporal and parietal, respectively. Most of the defects were deep to galea zone. The mean diameter of defect was 24.28 ± 45.37 mm for all of the patients. The difference between diameter of defect in both groups was statistically meaningful, while no such difference between graft diameter was seen. The graft 'Take' was completely successful in both groups according to evaluations. The level of postoperative pain was lower in the case group compared to the control according to VAS scale, and the satisfaction was higher in them per Likert scale. Conclusion: Scalp can safely be used as donor site for skin graft to be used for scalp defects, which is associated with better results and lower complication rates compared to other donor sites.

Keywords: donor site, leg, partial-thickness graft, scalp

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Authors: Ana Beltran Sanahuja, A. Valdés García, Saray Lopez De Pablo Gallego, Maria Soledad Prats Moya

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Tomato consumption has greatly increased worldwide in the last few years, mostly due to a growing demand for products like sofrito. In this sense, regular consumption of tomato-based products has been consistently associated with a reduction in the incidence of chronic degenerative diseases. The sofrito is a homemade tomato sauce typical of the Mediterranean area, which contains as main ingredients: tomato, onion, garlic and olive oil. There are also sofrito’s variations by adding other spices which bring at the same time not only color, flavor, smell and or aroma; they also provide medicinal properties, due to their antioxidant power. This protective effect has mainly been attributed to the predominant bioactive compounds present in sofrito, such as lycopene and other carotenoids as well as more than 40 different polyphenols. Regarding the cooking process, it is known that it can modify the properties and the availability of nutrients in sofrito; however, there is not enough information regarding this issue. For this reason, the aim of the present work is to evaluate the cooking effect on the antioxidant capacity of different variants of tomato-based sofrito combined with other spices, through the analysis of total phenols content (TPC) and to evaluate the antioxidant capacity by using the method of free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH). Based on the results obtained, it can be confirmed that the basic sofrito composed of tomato, onion, garlic and olive oil and the sofrito with 1 g of rosemary added, are the ones with the highest content of phenols presenting greater antioxidant power than other industrial sofrito, and that of other variables of sofrito with added thyme or higher amounts of garlic. Moreover, it has been observed that in the elaboration of the tomato-based sofrito, it is possible to cook until 60 minutes, since the cooking process increases the bioavailability of the carotenoids when breaking the cell walls, which weakens the binding forces between the carotenoids and increases the levels of antioxidants present, confirmed both with the TPC and DPPH methods. It can be concluded that the cooking process of different variants of tomato-based sofrito, including spices, can improve the antioxidant capacity. The synergistic effects of different antioxidants may have a greater protective effect; increasing, also, the digestibility of proteins. In addition, the antioxidants help to deactivate the free radicals of diseases such as atherosclerosis, aging, immune suppression, cancer, and diabetes.

Keywords: antioxidants, cooking process, phenols sofrito

Procedia PDF Downloads 141

Authors: P. Renjitha, P. Hari Prakash

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Auditory neuropathy spectrum disorder (ANSD) is an auditory disorder with normal cochlear outer hair cell function and disrupted auditory nerve function. It results in unique clinical characteristic with absent auditory brainstem response (ABR), absent acoustic reflex and the presence of otoacoustic emissions (OAE) and cochlear microphonics. The lesion site could be at cochlear inner hair cells, the synapse between the inner hair cells and type I auditory nerve fibers, and/or the auditory nerve itself. But the literatures on synchrony at higher auditory system are sporadic and are less understood. It might be interesting to see if there is a recovery of neural synchrony at higher auditory centers. Also, does the level at which the auditory system recovers with adequate synchrony to the extent of observable evoke response potentials (ERPs) can predict speech perception? In the current study, eight ANSD participants and healthy controls underwent detailed audiological assessment including ABR, auditory middle latency response (AMLR), and auditory late latency response (ALLR). AMLR was recorded for clicks and ALLR was evoked using 500Hz and 2 kHz tone bursts. Analysis revealed that the participant could be categorized into three groups. Group I (2/8) where ALLR was present only for 2kHz tone burst. Group II (4/8), where AMLR was absent and ALLR was seen for both the stimuli. Group III (2/8) consisted individuals with identifiable AMLR and ALLR for all the stimuli. The highest speech identification sore observed in ANSD group was 30% and hence considered having poor speech perception. Overall test result indicates that the site of neural synchrony recovery could be varying across individuals with ANSD. Some individuals show recovery of neural synchrony at the thalamocortical level while others show the same only at the cortical level. Within ALLR itself there could be variation across stimuli again could be related to neural synchrony. Nevertheless, none of these patterns could possible explain the speech perception ability of the individuals. Hence, it could be concluded that neural synchrony as measured by evoked potentials could not be a good clinical predictor speech perception.

Keywords: auditory late latency response, auditory middle latency response, auditory neuropathy spectrum disorder, correlation with speech identification score

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Authors: Kamila Dus-Szachniewicz, Artur Bednarkiewicz, Katarzyna Gdesz-Birula, Slawomir Drobczynski

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In modern oncology hyperthermia (HT) is defined as a controlled tumor heating. HT treatment temperatures range between 40–48 °C and can selectively damage heat-sensitive cancer cells or limit their further growth, usually with minimal injury to healthy tissues. Despite many advantages, conventional whole-body and regional hyperthermia have clinically relevant side effects, including cardiac and vascular disorders. Additionally, the lack of accessibility of deep-seated tumor sites and impaired targeting micrometastases renders HT less effective. It is believed that above disadvantages can significantly overcome by the application of biofunctionalized microparticles, which can specifically target tumor sites and become activated by an external stimulus to provide a sufficient cellular response. In our research, the unique optical tweezers system have enabled capturing the silica microparticles, primary cells and tumor spheroids in highly controllable and reproducible environment to study the impact of localized heat stimulation on normal and pathological cell and within multicellular tumor spheroid. High throughput spheroid model was introduced to better mimic the response to HT treatment on tumors in vivo. Additionally, application of local heating of tumor spheroids was performed in strictly controlled conditions resembling tumor microenvironment (temperature, pH, hypoxia, etc.), in response to localized and nonhomogeneous hyperthermia in the extracellular matrix, which promotes tumor progression and metastatic spread. The lack of precise control over these well- defined parameters in basic research leads to discrepancies in the response of tumor cells to the new treatment strategy in preclinical animal testing. The developed approach enables also sorting out subclasses of cells, which exhibit partial or total resistance to therapy, in order to understand fundamental aspects of the resistance shown by given tumor cells in response to given therapy mode and conditions. This work was funded by the National Science Centre (NCN, Poland) under grant no. UMO-2017/27/B/ST7/01255.

Keywords: cancer spheroids, hyperthermia, microparticles, optical tweezers

Procedia PDF Downloads 133

Authors: Ivna Mororó, Lise P. Labéjof, Stephanie Ribeiro, Kely Almeida

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Lipid droplets (LDs) are normally presented in greater or lesser number in the cytoplasm of almost all eukaryotic and some prokaryotic cells. They are independent organelles composed of a lipid ester core and a surface phospholipid monolayer. As a lipid storage form, they provide an available source of energy for the cell. Recently it was demonstrated that they play an important role in other many cellular processes. Among the many unresolved questions about them, it is not even known how LDs is formed, how lipids are recruited to LDs and how they interact with the other organelles. Excess fat in the organism is pathological and often associated with the development of some genetic, hormonal or behavioral diseases. The formation and accumulation of lipid droplets in the cytoplasm can be increased by exogenous physical or chemical agents. It is well known that ionizing radiation affects lipid metabolism resulting in increased lipogenesis in cells, but the details of this process are unknown. To better understand the mode of formation of LDs in liver cells, we investigate their ultrastructural morphology after irradiation. For that, Wistar rats were exposed to whole body gamma radiation from 60-cobalt at various single doses. Samples of the livers were processed for analysis under a conventional transmission electron microscope. We found that when compared to controls, morphological changes in liver cells were evident at the higher doses of radiation used. It was detected a great number of lipid droplets of different sizes and homogeneous content and some of them merged each other. In some cells, it was observed diffused LDs, not limited by a monolayer of phospholipids. This finding suggests that the phospholipid monolayer of the LDs was disrupted by ionizing radiation exposure that promotes lipid peroxydation of endo membranes. Thus the absence of the phospholipid monolayer may prevent the realization of some cellular activities as follow: - lipid exocytosis which requires the merging of LDs membrane with the plasma membrane; - the interaction of LDs with other membrane-bound organelles such as the endoplasmic reticulum (ER), the golgi and mitochondria and; - lipolysis of lipid esters contained in the LDs which requires the presence of enzymes located in membrane-bound organelles as ER. All these impediments can contribute to lipid accumulation in the cytoplasm and the development of diseases such as liver steatosis, cirrhosis and cancer.

Keywords: radiobiology, hepatocytes, lipid metabolism, transmission electron microscopy

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Authors: Wayde Veldman, Ozlem T. Bishop, Igor Polikarpov

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In bacteria, mono and disaccharides are phosphorylated during uptake into the cell via the widely used phosphoenolpyruvate (PEP)-dependent phosphotransferase transport system. As an initial step in the phosphorylated disaccharide metabolism pathway, certain glycoside hydrolase family 1 (GH1) enzymes play a crucial role in releasing phosphorylated and non-phosphorylated monosaccharides. However, structural determinants for the specificity of these enzymes still need to be clarified. GH1 enzymes are known to have a wide array of functions. According to the CAZy database, there are twenty-one different enzymatic activities in the GH1 family. Here, the structure and substrate specificity of a GH1 enzyme from Bacillus licheniformis, hereafter known as BlBglH, was investigated. The sequence of the enzyme BlBglH was compared to the sequences of other characterized GH1 enzymes using sequence alignment, sequence identity calculations, phylogenetic analysis, and motif discovery. Through these various analyses, BlBglH was found to have sequence features characteristic of the 6-phospho-β-glucosidase activity enzymes. Additionally, motif and structure comparisons of the three most commonly studied GH1 enzyme-activities revealed a shared loop amongst the different structures that consist of different sequence motifs – this loop is thought to guide specific substrates (depending on activity) towards the active-site. To further affirm BlBglH enzyme activity, molecular docking and molecular dynamics simulations were performed. Docking was carried out using 6-phospho-β-glucosidase enzyme-activity positive (p-Nitrophenyl-beta-D-glucoside-6-phosphate) and negative (p-Nitrophenyl-beta-D-galactoside-6-phosphate) control ligands, followed by 400 ns molecular dynamics simulations. The positive-control ligand maintained favourable interactions within the active site until the end of the simulation. The negative-control ligand was observed exiting the enzyme at 287 ns. Binding free energy calculations showed that the positive-control complex had a substantially more favourable binding energy compared to the negative-control complex. Jointly, the findings of this study suggest that the BlBglH enzyme possesses 6-phospho-β-glucosidase enzymatic activity.

Keywords: 6-P-β-glucosidase, glycoside hydrolase 1, molecular dynamics, sequence analysis, substrate specificity

Procedia PDF Downloads 130

Authors: Liang Wang, Wei Zhang

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Nowadays, sustainable development has increasingly become an important research topic of engineering education. Existing research on Engineering Education for Sustainable Development (EESD) has highlighted the importance of perceptions for ethical responsibility to address sustainable development in practice. However, whether and how the professional engineering experience affects those perceptions has not been proved, especially in a Chinese context. Our study fills this gap by investigating the perceptions bias of EESD between experienced engineers and engineering students. We specifically examined what EESD means for experienced engineers and engineering students using a triple-dimensional model to understand if there are obvious differences between the two groups. Our goal is to make the benefits of these experiences more accessible in school context. The data (n=438) came from a questionnaire created and adapted from previously published studies containing 288 students from mechanical or civil engineering and 150 civil engineers with rich working experience, and the questionnaire was distributed during Fall 2020. T-test was used to find the difference in different dimensions between the two groups. The statistical results show that there is a significant difference in the perceptions of EESD between experienced engineers and inexperienced engineering students in China. Experienced engineers tend to consider sustainable development from ecological, economic, and social perspectives, while engineering students' answers focus more on ecology and ignore economic and social dimensions to some extend. The findings provide empirical evidence that professional experience is helpful to cultivate the cognition and ability of sustainable development in engineering education. The results of this work indicate that more practical content should be added to engineering education to promote sustainable development. In addition, for the design of engineering courses and professional practice systems for sustainable development, we should not only pay attention to the ecological aspects but also emphasize the coordination of ecological, economic, and socially sustainable development (e.g., engineer's ethical responsibility).

Keywords: engineering education, sustainable development, experienced engineers, engineering students

Procedia PDF Downloads 102

Authors: Caroline M. Spagnol, Vera L. B. Isaac, Marcos A. Corrêa, Hérida R. N. Salgado

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Phenolic compounds are abundant in the Brazilian plant kingdom and they are part of a large and complex group of organic substances. Cinnamic acids are part of this group of organic compounds, and caffeic acid (CA) is one of its representatives. Antioxidants are compounds which act as free radical scavengers and, in other cases, such as metal chelators, both in the initiation stage and the propagation of oxidative process. The tyrosinase, polyphenol oxidase, is an enzyme that acts at various stages of melanin biosynthesis within the melanocytes and is considered a key molecule in this process. Some phenolic compounds exhibit inhibitory effects on melanogenesis by inhibiting the tyrosinase enzymatic activity and therefore has been the subject of studies. However, few studies have reported the effectiveness of these products and their safety. Objectives: To assess the inhibitory activity of tyrosinase, the antioxidant activity of CA and its cytotoxic potential. The method to evaluate the inhibitory activity of tyrosinase aims to assess the reduction transformation of L-dopa into dopaquinone reactions catalyzed by the enzyme. For evaluating the antioxidant activity was used the analytical methodology of DPPH radical inhibition. The cytotoxicity evaluation was carried out using the MTT method (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide), a colorimetric assay which determines the amount of insoluble violet crystals formed by the reduction of MTT in the mitochondria of living cells. Based on the results obtained during the study, CA has low activity as a depigmenting agent. However, it is a more potent antioxidant than ascorbic acid (AA), since a lower amount of CA is sufficient to inhibit 50% of DPPH radical. The results are promising since CA concentration that promoted 50% toxicity in HepG2 cells (IC50=781.8 μg/mL) is approximately 330 to 400 times greater than the concentration required to inhibit 50% of DPPH (IC50 DPPH= 2.39 μg/mL) and ABTS (IC50 ABTS= 1.96 μg/mL) radicals scavenging activity, respectively. The maximum concentration of caffeic acid tested (1140 mg /mL) did not reach 50% of cell death in HaCat cells. Thus, it was concluded that the caffeic acid does not cause toxicity in HepG2 and HaCat cells in the concentrations required to promote antioxidant activity in vitro, and it can be applied in topical products.

Keywords: caffeic acid, antioxidant, cytotoxicity, cosmetic

Procedia PDF Downloads 379

Authors: Ming Lu, Xiaojun Li, Bodi Lu, Juehui Xing

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Earthquake damage phenomenon of each large earthquake gives comprehensive and profound real test to the dynamic performance and failure mechanism of different engineering structures. Cognitive engineering structure characteristics through seismic damage phenomenon are often far superior to expensive shaking table experiments. After the earthquake, people will record a variety of different types of engineering damage photos. However, a large number of earthquake damage photographs lack sufficient information and reduce their using value. To improve the research value and the use efficiency of engineering seismic damage photographs, this paper objects to explore and show seismic damage background information, which includes the earthquake magnitude, earthquake intensity, and the damaged structure characteristics. From the research requirement in earthquake engineering field, the authors use the 2008 China Wenchuan M8.0 earthquake photographs, and provide four kinds of attribute indexes and classification, which are seismic information, structure types, earthquake damage parts and disaster causation factors. The final object is to set up an engineering structural seismic damage database based on these four attribute indicators and classification, and eventually build a website providing seismic damage photographs.

Keywords: attribute index, classification method, earthquake damage picture, engineering structure

Procedia PDF Downloads 765

Authors: Vinko Lešić, Mario Vašak, Anita Martinčević, Marko Gulin, Antonio Starčić, Hrvoje Novak

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With 40% of total world energy consumption, building systems are developing into technically complex large energy consumers suitable for application of sophisticated power management approaches to largely increase the energy efficiency and even make them active energy market participants. Centralized control system of building heating and cooling managed by economically-optimal model predictive control shows promising results with estimated 30% of energy efficiency increase. The research is focused on implementation of such a method on a case study performed on two floors of our faculty building with corresponding sensors wireless data acquisition, remote heating/cooling units and central climate controller. Building walls are mathematically modeled with corresponding material types, surface shapes and sizes. Models are then exploited to predict thermal characteristics and changes in different building zones. Exterior influences such as environmental conditions and weather forecast, people behavior and comfort demands are all taken into account for deriving price-optimal climate control. Finally, a DC microgrid with photovoltaics, wind turbine, supercapacitor, batteries and fuel cell stacks is added to make the building a unit capable of active participation in a price-varying energy market. Computational burden of applying model predictive control on such a complex system is relaxed through a hierarchical decomposition of the microgrid and climate control, where the former is designed as higher hierarchical level with pre-calculated price-optimal power flows control, and latter is designed as lower level control responsible to ensure thermal comfort and exploit the optimal supply conditions enabled by microgrid energy flows management. Such an approach is expected to enable the inclusion of more complex building subsystems into consideration in order to further increase the energy efficiency.

Keywords: price-optimal building climate control, Microgrid power flow optimisation, hierarchical model predictive control, energy efficient buildings, energy market participation

Procedia PDF Downloads 465