Search results for: interferon gamma release assay
1337 Official Game Account Analysis: Factors Influence Users' Judgments in Limited-Word Posts
Authors: Shanhua Hu
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Social media as a critical propagandizing form of film, video games, and digital products has received substantial research attention, but there exists several critical barriers such as: (1) few studies exploring the internal and external connections of a product as part of the multimodal context that gives rise to readability and commercial return; (2) the lack of study of multimodal analysis in product’s official account of game publishers and its impact on users’ behaviors including purchase intention, social media engagement, and playing time; (3) no standardized ecologically-valid, game type-varying data can be used to study the complexity of official account’s postings within a time period. This proposed research helps to tackle these limitations in order to develop a model of readability study that is more ecologically valid, robust, and thorough. To accomplish this objective, this paper provides a more diverse dataset comprising different visual elements and messages collected from the official Twitter accounts of the Top 20 best-selling games of 2021. Video game companies target potential users through social media, a popular approach is to set up an official account to maintain exposure. Typically, major game publishers would create an official account on Twitter months before the game's release date to update on the game's development, announce collaborations, and reveal spoilers. Analyses of tweets from those official Twitter accounts would assist publishers and marketers in identifying how to efficiently and precisely deploy advertising to increase game sales. The purpose of this research is to determine how official game accounts use Twitter to attract new customers, specifically which types of messages are most effective at increasing sales. The dataset includes the number of days until the actual release date on Twitter posts, the readability of the post (Flesch Reading Ease Score, FRES), the number of emojis used, the number of hashtags, the number of followers of the mentioned users, the categorization of the posts (i.e., spoilers, collaborations, promotions), and the number of video views. The timeline of Twitter postings from official accounts will be compared to the history of pre-orders and sales figures to determine the potential impact of social media posts. This study aims to determine how the above-mentioned characteristics of official accounts' Twitter postings influence the sales of the game and to examine the possible causes of this influence. The outcome will provide researchers with a list of potential aspects that could influence people's judgments in limited-word posts. With the increased average online time, users would adapt more quickly than before in online information exchange and readings, such as the word to use sentence length, and the use of emojis or hashtags. The study on the promotion of official game accounts will not only enable publishers to create more effective promotion techniques in the future but also provide ideas for future research on the influence of social media posts with a limited number of words on consumers' purchasing decisions. Future research can focus on more specific linguistic aspects, such as precise word choice in advertising.Keywords: engagement, official account, promotion, twitter, video game
Procedia PDF Downloads 761336 Optimization of Agricultural Water Demand Using a Hybrid Model of Dynamic Programming and Neural Networks: A Case Study of Algeria
Authors: M. Boudjerda, B. Touaibia, M. K. Mihoubi
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In Algeria agricultural irrigation is the primary water consuming sector followed by the domestic and industrial sectors. Economic development in the last decade has weighed heavily on water resources which are relatively limited and gradually decreasing to the detriment of agriculture. The research presented in this paper focuses on the optimization of irrigation water demand. Dynamic Programming-Neural Network (DPNN) method is applied to investigate reservoir optimization. The optimal operation rule is formulated to minimize the gap between water release and water irrigation demand. As a case study, Foum El-Gherza dam’s reservoir system in south of Algeria has been selected to examine our proposed optimization model. The application of DPNN method allowed increasing the satisfaction rate (SR) from 12.32% to 55%. In addition, the operation rule generated showed more reliable and resilience operation for the examined case study.Keywords: water management, agricultural demand, dam and reservoir operation, Foum el-Gherza dam, dynamic programming, artificial neural network
Procedia PDF Downloads 1151335 The Role of Txnrd2 Deficiency in Epithelial-to-Mesenchymal-Transition (EMT) and Tumor Formation in Pancreatic Cancer
Authors: Chao Wu
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Thioredoxin reductase 2 is a mitochondrial enzyme that belongs to the cellular defense against oxidative stress. We deleted mitochondrial Txnrd2 in a KrasG12D-driven pancreatic tumor model. Despite an initial increase in precursor lesions, tumor incidence decreased significantly. We isolated cancer cell lines from these genetically engineered mice and observed an impaired proliferation and colony formation. Reactive Oxygen Species, as determined by DCF fluorescence, were increased. We detected a higher mitochondrial copy number in Txnrd2-deficient cells (KTP). However, measurement of mitochondrial bioenergetics showed no impairment of mitochondrial function and comparable O₂-consumption and extracellular acidification rates. In addition, the mitochondrial complex composition was affected in Txnrd2 deleted cell lines. To gain better insight into the role of Txnrd2, we deleted Txnrd2 in clones from parental KrasG12D cell lines using Crispr/Cas9 technology. The deletion was confirmed by western blot and activity assay. Interestingly, and in line with previous RNA expression analysis, we saw changes in EMT markers in Txnrd2 deleted cell lines and control cell lines. This might help us explain the reduced tumor incidence in KrasG12D; Txnrd2∆panc mice.Keywords: PDAC, TXNRD2, epithelial-to-mesenchymal-transition, ROS
Procedia PDF Downloads 1221334 Phytochemical Profile and in Vitro Bioactivity Studies on Two Underutilized Vegetables in Nigeria
Authors: Borokini Funmilayo Boede
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B. alba L., commonly called ‘Amunututu’ and Solanecio biafrae called ‘Worowo’ among the Yoruba tribe in the southwest part of Nigeria are reported to be of great ethnomedicinal importance but are among many underutilized green leafy vegetables in the country. Many studies have established the nutritional values of these vegetables, utilization are very poor and indepth information on their chemical profiles is scarce. The aqueous, methanolic and ethanolic extracts of these vegetables were subjected to phytochemical screening and phenolic profiles of the alcoholic extracts were characterized by using high-performance liquid chromatography coupled with diode array detector (HPLC-DAD). Total phenol and flavonoid contents were determined, antioxidant activities were evaluated using five in vitro assays to assess DPPH, nitric oxide and hydroxyl radical-scavenging abilities, as well as reducing power with ferric reducing antioxidant assay and phosphomolybdate method. The antibacterial activities of the extracts against Staphylococcus aureus, Pseudomonas aeruginosa, and Salmonella typhi were evaluated by using agar well diffusion method and the antifungal activity evaluated against food-associated filamentous fungi by using poisoned food technique with the aim of assessing their nutraceutical potentials to encourage their production and utilization. The results revealed the presence of saponnin, steroids, tannin, terpenoid and flavonoid as well as phenolic compounds: gallic acid, chlorogenic acid, caffeic acid, coumarin, rutin, quercitrin, quercetin and kaemferol. The vegetables showed varying concentration dependent reducing and radical scavenging abilities from weak to strong compared with gallic acid, rutin, trolox and ascorbic acid used as positive controls; the aqueous extracts which gave higher concentrations of total phenol displayed higher ability to reduce Fe (lll) to Fe (ll) and stronger inhibiting power against hydroxyl radical than the alcoholic extracts and in most cases exhibited more potency than the ascorbic acids used as positive controls, at the same concentrations, whereas, methanol and / or ethanol extracts were found to be more effective in scavenging 2, 2-diphenyl-1-picryl hydrazyl radical and showed higher ability to reduce Mo (VI) to Mo (V) in total antioxidant assay than the aqueous extracts. However, the inhibition abilities of all the extracts against nitric oxide were comparable with the ascorbic acid control at the same concentrations. There were strong positive correlations with total phenol (mg GAE/g) and total flavonoid (mg RE/g) contents in the range TFC (r=0.857- 0999 and r= 0.904-1.000) and TPC (r= 0.844- 0.992 and r= 0.900 -0.999) for Basella alba and Senecio biafrae respectively. Inhibition concentration at 50 % (IC50) for each extract to scavenge DPPH, OH and NO radicals ranged from 32.73 to 1.52 compared with control (0.846 - -6.42) mg/ml. At 0.05g/ml, the vegetables were found to exhibit mild antibacterial activities against Staphylococcus aureus, Pseudomonas aeruginosa and Salmonella typhi compared with streptomycin sulphate used as control but appreciable antifungi activities against (Trichoderma rubrum and Aspergillus fumigates) compared with bonlate antibiotic positive control. The vegetables possess appreciable antioxidant and antimicrobial properties for promoting good health, their cultivation and utilization should be encouraged especially in the face of increasing health and economic challenges and food insecurity in many parts of the world.Keywords: antimicrobial, antioxidants, extracts, phytochemicals
Procedia PDF Downloads 3241333 The Role of Moringa oleifera Extract Leaves in Inducing Apoptosis in Breast Cancer Cell Line
Authors: V. Yurina, H. Sujuti, E. Rahmani, A. R. Nopitasari
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Breast cancer has the highest prevalence cancer in women. Moringa leaves (M. oleifera) contain quercetin, kaempferol, and benzyl isothiocyanate which can enhance induction of apoptosis. This research aimed to study the role of the leaf extract of Moringa to increase apoptosis in breast cancer cell line, MCF-7 cells. This research used in vitro experimental, post-test only, control group design on breast cancer cells MCF-7 in vitro. Moringa leaves were extracted by maceration method with ethanol 70%. Cells were treated with drumstick leaves extract on 1100, 2200, and 4400 μg/ml for Hsp27 and caspase-9 expression (immunocytochemistry) and apoptosis (TUNEL assay) test. The results of this study found that the IC50 2200 µg/ml. Moringa leaves extract can significantly increase the expression of caspase-9 (p<0.05) and decreased Hsp 27 expression (p<0.05). Moreover it can increase apoptosis (p<0.05) significantly in MCF-7 cells. The conclusion of this study is Moringa leaves extract is able to increase the expression of caspase-9, decrease Hsp27 expression and increase apoptosis in breast cancer cell-line MCF-7.Keywords: apoptosis, breast cancer, caspase-9, Hsp27, Moringa oleifera
Procedia PDF Downloads 5441332 The Effects of Fertilizer in the Workplace on Male Infertility: About Workers of Unit NPK in Complex Fertial Annaba
Authors: B. Loukil, L. Mallem, M. S. Boulakoud
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Inorganic fertilizers consist mainly of salts of ammonium nitrate, phosphate and potassium, the combination of primary nutrients NPK including secondary and micro nutrients are essential for plant growth, used for intensive agriculture, ranching, and horticultural crops, to increase soil fertility and ensure sustainable crop production. The manufacture of fertilizers is generally at a high temperature and high pressure, in the presence of several highly hazardous chemicals, dust and gases. These products are absorbed high in the airway, increasing the airway resistance thereby adversely affecting the pulmonary functions of workers. A study was conducted on 34 employees, especially exposed to nitrate derivatives. A questionnaire was prepared and distributed to all employees in the unit. The workers were divided into two groups according to age. Several hormonal parameters Assay were measured. The results of the questionnaire have detected a fertility problem, Concerning the hormones a significant reduction in the concentration of testosterone in both groups and LH in the group aged 30 to 40 year were noted compared to the control. However, an increase in the concentration of prolactin in both groups compared to the control. There was a significant decrease in FSH in the group aged 30 to 40 always in compared with the control group.Keywords: fertilizers, healthy worker, risk, fertility
Procedia PDF Downloads 3991331 Determination of Aflatoxins in Edible-Medicinal Plant Samples by HPLC with Fluorescence Detector and KOBRA-Cell
Authors: Isil Gazioglu, Abdulselam Ertas
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Aflatoxins (AFs) are secondary toxic metabolites of Aspergillus flavus and A. parasiticus. AFs can be absorbed through the skin. Potent carcinogens like AFs should be completely absent from cosmetics, this can be achieved by careful quality control of the raw plant materials. Regulatory limits for aflatoxins have been established in many countries, and reliable testing methodology is needed to implement and enforce the regulatory limits. In this study, ten medicinal plant samples (Bundelia tournefortti, Capsella bursa-pastoris, Carduus tenuiflorus, Cardaria draba, Malva neglecta, Malvella sharardiana, Melissa officinalis, Sideritis libanotica, Stakys thirkei, Thymus nummularius) were investigated for aflatoxin (AF) contaminations by employing an HPLC assay for the determination of AFB1, B2, G1 and G2. The samples were extracted with 70% (v/v) methanol in water before further cleaned up with an immunoaffinity column and followed by the detection of AFs by using an electrochemically post-column derivatization with Kobra-Cell and fluorescence detector. The extraction procedure was optimized in order to obtain the best recovery. The method was successfully carried out with all medicinal plant samples. The results revealed that five (50%) of samples were contaminated with AFs. The association between particular samples and the AF contaminated could not be determined due to the low frequency of positive samples.Keywords: aflatoxin B1, HPLC-FLD, KOBRA-Cell, mycotoxin
Procedia PDF Downloads 6051330 Management of H. Armigera by Using Various Techniques
Authors: Ajmal Khan Kassi, Humayun Javed, Syed Abdul Qadeem
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The study was conducted to find out the best management practices against American bollworm on Okra variety Arka Anamika during 2016. The three different management practices viz. Release of Trichogramma chilonis, hoeing and weeding, clipping and lufenuron insect growth regulator (IGR) which were tested individually and with all possible combinations for the controlling of American bollworm at 3 diverse areas viz. University Research Farm Koont, NARC and Farmer Field Taxila. All the treatment combinations regarding damage of fruit showed significant results. The minimum fruit infestation i.e. 3.20% and 3.58% was recorded with combined treatment (i.e. T. chilonis + hoeing + weeding + lufenuron) in two different localities. This combined treatment also resulted in maximum yield at NARC and Taxila i.e. 57.67 and 62.66 q/ha respectively. This treatment gave the best results to manage H. armigera. On the basis of different integrated pest management techniques, Arka Anamika variety proved to be comparatively resistant against H. armigera in different localities. So this variety is recommended for the cultivation in Pothwar region to get maximum yield.Keywords: management, american bollworm, arka anamika, okra
Procedia PDF Downloads 551329 UV-Reactive Electrospinning: Preparation, Characterization and Cell Culture Applications of Nanofiber Scaffolds Containing Keratin
Authors: Duygu Yüksel Deniz, Memet Vezir Kahraman, Serap Erdem Kuruca, Mediha Süleymanoğlu
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Our first aim was to synthesize Hydroxy Apatite (HAP) and then modify its surface by adding 4-Vinylbenzene boronic acid (4-VBBA). The characterization was done by FT-IR. By adding Polyvinyl alcohol (PVA) to 4- VBBA-HAP, we obtained a suitable electrospinning solution. PVA solution which was also modified by using alkoxy silanes, in order to prevent the scaffolds from being damaged by aqueous cell medium, was added. Keratin was dissolved and then added into the electrospinning solution. Keratin containing 4-VBBA- HAP/PVA composite was used to fabricate nanofiber scaffolds with the simultaneous UV-reactive electrospinning technique. The structural characterization was done by FT-IR. Thermal gravimetric analysis was also performed by using TGA. The morphological characterization was determined by SEM analyses. Our second aim was to create a scaffold where cells could grow. With this purpose, suitable nanofibers were choosen according to their SEM analysis. Keratin containing nanofibers were seeded with 3T3, ECV and SAOS cells and their cytotoxicity and cell proliferation were investigated by using MTT assay. After cell culturing process morphological characterization was determined by SEM analyses. These scaffolds were designed to be nontoxic biomaterials. Here, a comparision was made between keratin containing 3T3, ECV and SAOS seeded nanofiber scaffolds and the results were presented and discussed.Keywords: cell culture, keratin, nanofibers, UV-reactive electrospinning
Procedia PDF Downloads 4541328 A Prenylflavanoid, HME5 with Antiproliferative Activity in Human Ovarian Cancer Cells
Authors: Mashitoh Abd Rahman, Najihah Mohd Hashim, Faiqah Ramli, Syam Mohan, Noraziah Nordin, Hamed Karimian, Hapipah Mohd Ali
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Ovarian cancer is the most lethal gynecological malignancies. HME5, a prenylflavanoid has been isolated from local medicinal plant. This compound has been reported to possess a broad spectrum of biological activities including anticancer property. However, the potential of HME5 as an antiproliferative and cytotoxic agent on an ovarian cancer cells has not yet been investigated. In this present study, we examined the antiproliferative and cytotoxic effect of HME5 on Caov-3 (Human Ovarian Adenocarcinoma) cell line by using 3-[4,5-dimethylthizol-2-y]-2,5-diphenyltetrazolium bromide (MTT) assay, Acridine orange and propidium Iodide (AOPi) and cell cycle analysis study. HME5 has shown to inhibit Caov-3 in a time-dependent manner with the IC50 values of 5µg/ml, 2µg/ml and 1µg/ml after 24h, 48h and 72h treatment, respectively. Morphological study from AOPi analysis showed that HME5 induced apoptosis after 24 and 48h post-treatment. Nevertheless, HME5 exhibited cell cycle arrest at G1 phase as indicated in flow cytometry cell cycle profiling. In conclusion, HME5 inhibited proliferation of Caov-3 through induction of apoptosis and cell cycle arrest at G1 phase.Keywords: apoptosis, prenylflavanoid, ovarian cancer, HME5
Procedia PDF Downloads 4611327 Antibiotic Potential of Bioactive Compounds from a Marine Streptomyces Isolated from South Pacific Sediments
Authors: Ilaisa Kacivakanadina, Samson Viulu, Brad Carte, Katy Soapi
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Two bioactive compounds namely Vulgamycin (also known as enterocin A) and 5-deoxyenterocin were purified from a marine bacterial strain 1903. Strain 1903 was isolated from marine sediments collected from the Solomon Islands. Morphological features of strain 1903 showed that it belongs to the genus Streptomyces. The two secondary metabolites were extracted using EtOAc and purified by chromatographic methods using EtOAc and hexane solvents. Mass spectrum and NMR data of pure compounds were used to elucidate the chemical structures. In this study, results showed that both compounds were strongly active against Wild Type Staphylococcus aureus (WTSA) (MIC < 1 µg/mL) and in Brine shrimp assays (BSA) (MIC < 1 µg/mL). 5-deoxyenterocin was also active against Rifamycin resistant Staphylococcus aureus (RRSA) (MIC, 250 µg/mL) while vulgamycin showed bioactivity against Methicillin resistant Staphylococcus aureus (MRSA) (MIC 250 µg/mL). To the best of our knowledge, this is the first study that showed the bio-activity of 5-deoxyenterocin. This is also the first time that Vulgamycin has been reported to be active in a BSA. There has not been any mechanism of action studies for these two compounds against pathogens. This warrants further studies on their mechanism of action against microbial pathogens.Keywords: 5-deoxyenterocin, bioactivity, brine shrimp assay (BSA), vulgamycin
Procedia PDF Downloads 1891326 Unconventional Strategies for Combating Multidrug Resistant Bacterial Biofilms
Authors: Soheir Mohamed Fathey
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Biofilms are complex biological communities which are hard to be eliminated by conventional antibiotic administration and implemented in eighty percent of humans infections. Green remedies have been used for centuries and have shown obvious effects in hindering and combating microbial biofilm infections. Nowadays, there has been a growth in the number of researches on the anti-biofilm performance of natural agents such as plant essential oil (EOs) and propolis. In this study, we investigated the antibiofilm performance of various natural agents, including four essential oils (EOs), cinnamon (Cinnamomum cassia), tea tree (Melaleuca alternifolia), and clove (Syzygium aromaticum), as well as propolis versus the biofilm of both Gram-positive pathogenic bacterium Staphylococcus aureus and Gram-negative pathogenic bacterium Pseudomonas aeruginosa which are major human and animal pathogens rendering a high risk due to their biofilm development ability. The antibiofilm activity of the tested agents was evaluated by crystal violet staining assay and detected by scanning electron and fluorescent microscopy. Antibiofilm performance declared a potent effect of the tested products versus the tested bacterial biofilms.Keywords: biofilm, essential oils, electron microscopy, fluorescent
Procedia PDF Downloads 961325 Bioprotective Role of Soil Borne Bacillus Strains against Selected Fungal Pathogens of Agriculture Relevance
Authors: Asad Ali, Asif Jamal
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The agriculture productivity losses due to microbial pathogens have been a serious issue in Pakistan and rest of the world. Present work was designed to isolate soil borne microorganisms having the antagonistic ability against notorious phytopathogens. From the initial collection of 23 bacterial isolates, two potent strains of Bacillus were screened on the basis of their comparative efficacy against devastating fungal pathogens. The strains AK-1 and AK-5 showed excellent inhibitory indexes against the majority of tested fungal strains. It was noted that both strains of Bacillus showed significant biocontrolling activity against Aspergillus flavus, Fusarium moniliforme, Colletotricum falcatum, Botrytis cinerea, Aspergillus niger, Fusarium oxysporum, Phythopthora capsici and Rhizopus oryzae. The strain AK-1 was efficient to suppress Aspergillus species and Rhizopus oryzae while AK-5 expressed significant antagonistic activity against Fusarium, Botrytis and Colletotricum species. On the basis of in vitro assay, it can be postulated that the Bacillus strains AK-1 and AK-5 can be used as bio-protective agent against various plant diseases. In addition, their applications as natural pesticides could be very helpful to prevent the adverse effects of chemical pesticides.Keywords: bacillus species, biocontrol agent, biopesticides, phytopathogens
Procedia PDF Downloads 2411324 Biodegradation of Chlorpyrifos in Real Wastewater by Acromobacter xylosoxidans SRK5 Immobilized in Calcium Alginate
Authors: Saira Khalid, Imran Hashmi
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Agrochemical industries produce huge amount of wastewater containing pesticides and other harmful residues. Environmental regulations make it compulsory to bring pesticides to a minimum level before releasing wastewater from industrial units.The present study was designed with the objective to investigate biodegradation of CP in real wastewater using bacterial cells immobilized in calcium alginate. Bacterial strain identified as Acromobacter xylosoxidans SRK5 (KT013092) using 16S rRNA nucleotide sequence analysis was used. SRK5 was immobilized in calcium alginate to make calcium alginate microspheres (CAMs). Real wastewater from industry having 50 mg L⁻¹ of CP was inoculated with free cells or CAMs and incubated for 96 h at 37˚C. CP removal efficiency with CAMs was 98% after 72 h of incubation, and no lag phase was observed. With free cells, 12h of lag phase was observed. After 96 h of incubation 87% of CP removal was observed when inoculated with free cells. No adsorption was observed on vacant CAMs. Phytotoxicity assay demonstrated considerable loss in toxicity. Almost complete COD removal was achieved at 96 h with CAMs. Study suggests the use of immobilized cells of SRK5 for bioaugmentation of industrial wastewater for CP degradation instead of free cells.Keywords: biodegradation, chlorpyrifos, immobilization, wastewater
Procedia PDF Downloads 1781323 Quantum Modelling of AgHMoO4, CsHMoO4 and AgCsMoO4 Chemistry in the Field of Nuclear Power Plant Safety
Authors: Mohamad Saab, Sidi Souvi
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In a major nuclear accident, the released fission products (FPs) and the structural materials are likely to influence the transport of iodine in the reactor coolant system (RCS) of a pressurized water reactor (PWR). So far, the thermodynamic data on cesium and silver species used to estimate the magnitude of FP release show some discrepancies, data are scarce and not reliable. For this reason, it is crucial to review the thermodynamic values related to cesium and silver materials. To this end, we have used state-of-the-art quantum chemical methods to compute the formation enthalpies and entropies of AgHMoO₄, CsHMoO₄, and AgCsMoO₄ in the gas phase. Different quantum chemical methods have been investigated (DFT and CCSD(T)) in order to predict the geometrical parameters and the energetics including the correlation energy. The geometries were optimized with TPSSh-5%HF method, followed by a single point calculation of the total electronic energies using the CCSD(T) wave function method. We thus propose with a final uncertainty of about 2 kJmol⁻¹ standard enthalpies of formation of AgHMoO₄, CsHMoO₄, and AgCsMoO₄.Keywords: nuclear accident, ASTEC code, thermochemical database, quantum chemical methods
Procedia PDF Downloads 1891322 Antioxidative Effect of Bauhinia acuminata Water Extract Consumption in Rat
Authors: Amornnat Thuppia, Pornrut Rabintossaporn, Suphaket Saenthaweesuk, Nuntiya Somparn
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The aim of this present study was to determine the antioxidant effects and its mechanism of aqueous leaves extract of Bauhinia acuminata (BA) in rat. The extract was screened for its phytochemical contents and antioxidant activity in vitro. Moreover, the extract was studied in rats to evaluate its effects in vivo. Rats were orally administered with the extract at the dose of 50, 100 and 200 mg/kg for 28 days. Phytochemical screening of plant extracts showed the presence of saponin, alkaloid, cardiac glycosides, flavonoids, tannin and steroid compounds. The extract contained phenolic compounds 53.36 ± 1.01 mg of gallic acid equivalents per gram BA extract. The free radical scavenging activity assessed by DPPH assay gave IC50 of 44.47 ± 2.83 µg/mL, which is relatively lower than that of BHT with IC50 of 12.34 ± 1.14µg/mL. In the animals, the extract was well tolerated by the animals throughout the 28 days of study as shown by normal serum levels AST, ALP, ALT, BUN and Cr as well as normal histology of liver and pancreatic and kidney tissue. Significantly, reduction of serum oxidative stress markers malondialdehyde (MDA) was found in rat treated with BA extract compared with control. Taken together, this study provides evidence that Bauhinia acuminata (BA) exhibits direct antioxidant properties and induces cytoprotective enzyme in vivo.Keywords: Bauhinia acuminata, antioxidant, malondialdehyde (MDA), oxidative marker
Procedia PDF Downloads 2731321 Potential Activities of Human Endogenous Retroviral kDNA in Melanoma Pathogenesis and HIV-1 Infection
Authors: Jianli Dong, Fangling Xu, Gengming Huang
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Human endogenous retroviral elements (HERVs) comprise approximately 8% of the human genome. They are thought to be germline-integrated genetic remnants of retroviral infections. Although HERV sequences are highly defective, some, especially the K type (HERV-K), have been shown to be expressed and may have biological activities in the pathogenesis of cancer, chronic inflammation and autoimmune diseases. We found that HERV-K GAG and ENV proteins were strongly expressed in pleomorphic melanoma cells. We also detected a critical role of HERV-K ENV in mediating intercellular fusion and colony formation of melanoma cells. Interestingly, we found that levels of HERV-K GAG and ENV expression correlated with the activation of ERK and loss of p16INK4A in melanoma cells, and inhibition of MEK or CDK4, especially in combination, reduced HERV-K expression in melanoma cells. We also performed a reverse transcription-polymerase chain reaction (RT-PCR) assay using DNase I digestion to remove “contaminating” HERV-K genomic DNA and examined HERV-K RNA expression in plasma samples from HIV-1 infected individuals. We found a covariation between HERV-K RNA expression and CD4 cell counts in HIV-1 positive samples. Although a causal link between HERV-K activation and melanoma development, and between HERV-K activation, HIV-1 infection and CD4 cell count have yet to be determined, existing data support the further research efforts in HERV-K.Keywords: CD4 cell, HERV-K, HIV-1, melanoma
Procedia PDF Downloads 2321320 The Mechanism of Antimicrobial Activity and Antioxidant Effects of the Essential Oil and the Methanolic Extract of Carum montanum (Coss. et Dur.) Benth. Et Hook. Aerial Parts from Algeria
Authors: Meriem El Kolli, Hocine Laouer, Hayet El Kolli, Salah Akkal
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The methanolic extract (ME) of C. montanum obtained by a hydo-alcoholic maceration and its polyphenol content was evaluated by Folin-Ciocalteu method. This extract and C. montanum essential oil were screened for antimicrobial activity against 21 microbial strains by agar diffusion method. MICs of the EO were determined by the broth micro dilution method. The mechanism of action of the EO was determined on the susceptible strains by the time kill assay and the lysis experience. Antioxidant properties were studied by both free DPPH radical scavenging and reducing power techniques. The TPC in the ME showed a high level of 101.50 ± 5.33 mg GAE /mg. B. cereus was the most sensitive strain with MIC of 55.5 µg/ml , then K. pneumoniae (111 µg/ml). A remarkable decrease in a survival rate as well as in the absorbance at 260 nm were recorded, which suggest that the cytoplasm membrane is one of the targets of the EO. Antioxidant effects were concentration dependent and IC50 values were 1.09 ± 0.37 µg/ml for the EO and 65.04 ± 0.00 µg/ml for the ME by DPPH method and a reducing power dose-dependent. In conclusion, C. montanum extracts showed potent which could be exploited in the food industry for food preservation.Keywords: C. montanum, Apiaceae, essential oils, antimicrobial activity, antioxidant activity, reducing power
Procedia PDF Downloads 2371319 Comparison Between a Droplet Digital PCR and Real Time PCR Method in Quantification of HBV DNA
Authors: Surangrat Srisurapanon, Chatchawal Wongjitrat, Navin Horthongkham, Ruengpung Sutthent
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HBV infection causes a potential serious public health problem. The ability to detect the HBV DNA concentration is of the importance and improved continuously. By using quantitative Polymerase Chain Reaction (qPCR), several factors in standardized; source of material, calibration standard curve and PCR efficiency are inconsistent. Digital PCR (dPCR) is an alternative PCR-based technique for absolute quantification using Poisson's statistics without requiring a standard curve. Therefore, the aim of this study is to compare the data set of HBV DNA generated between dPCR and qPCR methods. All samples were quantified by Abbott’s real time PCR and 54 samples with 2 -6 log10 HBV DNA were selected for comparison with dPCR. Of these 54 samples, there were two outlier samples defined as negative by dPCR. Of these two, samples were defined as negative by dPCR, whereas 52 samples were positive by both the tests. The difference between the two assays was less than 0.25 log IU/mL in 24/52 samples (46%) of paired samples; less than 0.5 log IU/mL in 46/52 samples (88%) and less than 1 log in 50/52 samples (96%). The correlation coefficient was r=0.788 and P-value <0.0001. Comparison to qPCR, data generated by dPCR tend to be the overestimation in the sample with low HBV DNA concentration and underestimated in the sample with high viral load. The variation in DNA by dPCR measurement might be due to the pre-amplification bias, template. Moreover, a minor drawback of dPCR is the large quantity of DNA had to be used when compare to the qPCR. Since the technology is relatively new, the limitations of this assay will be improved.Keywords: hepatitis B virus, real time PCR, digital PCR, DNA quantification
Procedia PDF Downloads 4811318 Molecular Detection and Characterization of Shiga Toxogenic Escherichia coli Associated with Dairy Product
Authors: Mohamed Al-Hazmi, Abdullah Al-Arfaj, Moussa Ihab
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Raw, unpasteurized milk can carry dangerous bacteria such as Salmonella, E. coli, and Listeria, which are responsible for causing numerous foodborne illnesses. The objective of this study was molecular characterization of shiga toxogenic E. coli in raw milk collected from different Egyptian governorates by multiplex PCR. During the period of 25th May to 25th October 2012, a total of 320 bulk-tank milk samples were collected from 10 cow farms located in different Egyptian governorates. Bacteriological examination of milk samples revealed the presence of E. coli organisms in 65 samples (20.3%), serotyping of the E. coli isolates revealed, 35 strains (10.94%) O111, 15 strains (4.69%) O157: H7, 10 strains (3.13%) O128 and 5 strains (1.56%) O119. Multiplex PCR for detection of shiga toxin type 2 and intimin genes revealed positive amplification of 255 bp fragment of shiga toxin type 2 gene and 384 bp fragment of intimin gene from all E. coli serovar O157: H7, while from serovar O111 were 25 (71.43%), 20 (57.14%) and from serovar O128 were 6 (60%), 8 (80%), respectively. The results of multiplex PCR assay are useful for identification of STEC possessing the eaeA and stx2 genes.Keywords: raw milk, E. coli, multiplex PCR, Shiga toxin type 2, intimin gene
Procedia PDF Downloads 3061317 Honey Contamination in the Republic of Kazakhstan
Authors: B. Sadepovich Maikanov, Z. Shabanbayevich Adilbekov, R. Husainovna Mustafina, L. Tyulegenovna Auteleyeva
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This study involves detailed information about contaminants of honey in the Republic of Kazakhstan. The requirements of the technical regulation ‘Requirements to safety of honey and bee products’ and GOST 19792-2001 were taken into account in this research. Contamination of honey by antibiotics wqs determined by the IEA (immune-enzyme analysis), Ridder analyzer and Tecna produced test systems. Voltammetry (TaLab device) was used to define contamination by salts of heavy metals and gamma-beta spectrometry, ‘Progress BG’ system, with preliminary ashing of the sample of honey was used to define radioactive contamination. This article pointed out that residues of chloramphenicol were detected in 24% of investigated products, in 22% of them –streptomycin, in 7.3% - sulfanilamide, in 2.4% - tylosin, and in 12% - combined contamination was noted. Geographically, the greatest degree of contamination of honey with antibiotics occurs in the Northern Kazakhstan – 54.4%, and Southern Kazakhstan - 50%, and the lowest in Central and Eastern Kazakhstan with 30% and 25%, respectively. Generally, pollution by heavy metals is within acceptable limits, but the contamination from lead is highest in the Akmola region. The level of radioactive cesium and strontium is also within acceptable concentrations. The highest radioactivity in terms of cesium was observed in the East Kazakhstan region - 49.00±10 Bq/kg, in Akmola, North Kazakhstan and Almaty - 12.00±5, 11.05±3 and 19.0±8 Bq/kg, respectively, while the norm is 100 Bq/kg. In terms of strontium, the radioactivity in the East Kazakhstan region is 25.03±15 Bq/kg, while in Akmola, North Kazakhstan and Almaty regions it is 12.00±3, 10.2±4 and 1.0±2 Bq/kg, respectively, with the norm of 80 Bq/kg. This accumulation is mainly associated with the environmental degradation, feeding and treating of bees. Moreover, in the process of collecting nectar, external substances can penetrate honey. Overall, this research determines factors and reasons of honey contamination.Keywords: antibiotics, contamination of honey, honey, radionuclides
Procedia PDF Downloads 2261316 In vitro Antioxidant and DNA Protectant Activity of Different Skin Colored Eggplant (Solanum melongena)
Authors: K. M. Somawathie, V. Rizliya, H. A. M. Wickrmasinghe, Terrence Madhujith
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The main objective of our study was to determine the in vitro antioxidant and DNA protectant activity of aqueous extract of S. melongena with different skin colors; dark purple (DP), moderately purple (MP), light purple (LP) and purple and green (PG). The antioxidant activity was evaluated using the DPPH and ABTS free radical scavenging assay, ferric reducing antioxidant power (FRAP), ferric thiocyanate (FTC) and the egg yolk model. The effectiveness of eggplant extracts against radical induced DNA damage was also determined. There was a significant difference (p < 0.0001) between the skin color and antioxidant activity. TPC and FRAP values of eggplant extracts ranged from 48.67±0.27-61.11±0.26 (mg GAE/100 g fresh weight) and 4.19±0.11-7.46±0.26 (mmol of FeS04/g of fresh weight) respectively. MP displayed the highest percentage of DPPH radical scavenging activity while, DP demonstrated the strongest total antioxidant capacity. In the FTC and egg yolk model, DP and MP showed better antioxidant activity than PG and LP. All eggplant extracts showed potent antioxidant activity in retaining DNA against AAPH mediated radical damage. DP and MP demonstrated better antioxidant activity which may be attributed to the higher phenolic content since a positive correlation was observed between the TPC and the antioxidant parameters.Keywords: Solanum melongena, skin color, antioxidant, DNA protection, lipid peroxidation
Procedia PDF Downloads 4311315 The Environmental Benefits of the Adoption of Emission Control for Locomotives in Brazil
Authors: Rui de Abrantes, André Luiz Silva Forcetto
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Air pollution is a big problem in many cities around the world. Brazilian big cities also have this problem, where millions of people are exposed daily to pollutants levels above the recommended by WHO. Brazil has taken several actions to reduce air pollution, among others, controlling the atmospheric emissions from vehicles, non-road mobile machinery, and motorcycles, but on the other side, there are no emissions controls for locomotives, which are exposing the population to tons of pollutants annually. The rail network is not homogeneously distributed in the national territory; it is denser near the big cities, and this way, the population is more exposed to pollutants; apart from that, the government intends to increase the rail network as one of the strategies for greenhouse gas mitigation, complying with the international agreements against the climate changes. This paper initially presents the estimated emissions from locomotive fleets with no emission control and with emission control equivalent to US Tier 3 from 2028 and for the next 20 years. However, we realized that a program equivalent to phase Tier 3 would not be effective, so we proposed a program in two steps that will avoid the release of more than 2.4 million tons of CO and 531,000 tons of hydrocarbons, 3.7 million tons of nitrogen oxides, and 102,000 tons of particulate matter in 20 years.Keywords: locomotives, emission control, air pollution, pollutants emission
Procedia PDF Downloads 491314 Isolation of Cytotoxic Compound from Tectona grandis Stem to Be Used as Thai Medicinal Preparation for Cancer Treatment
Authors: Onmanee Prajuabjinda, Pakakrong Thondeeying, Jipisute Chunthorng-Orn, Bhanuz Dechayont, Arunporn Itharat
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A Thai medicinal preparation has been used for cancer treatment more than ten years ago in Khampramong Temple. Tectona grandis stem is one ingredient of this Thai medicinal remedy. The ethanolic extract of Tectona grandis stem showed the highest cytotoxic activities against human breast adenocarcinoma (MCF-7), but was less cytotoxic against large cell lung carcinoma (COR-L23) (IC50 = 3.92 and 7.78 µg/ml, respectively). It was isolated by bioassay-guided isolation method. Tectoquinone, a anthraquinone compound was isolated from this plant. This compound showed high specific cytotoxicity against human breast adenocarcinoma (MCF-7), but was less cytotoxic against large cell lung carcinoma (COR-L23)(IC50 =16.15 and 47.56 µg/ml or 72.67 and 214.00 µM, respectively). However, it showed less cytotoxic activity than the crude extract. In conclusion, tectoquinone as a main compound, is not the best cytotoxic compound from Tectona grandis, so there are more active cytotoxic compounds in this extract which should be isolated in the future. Moreover, tectoquinone displayed specific cytotoxicity against only human breast adenocarcinoma (MCF-7) which is a good criterion for cancer treatment.Keywords: Tectona grandis, SRB assay, cytotoxicity, tectoquinone
Procedia PDF Downloads 4321313 Study of Fire Propagation and Soot Flow in a Pantry Car of Railway Locomotive
Authors: Juhi Kaushik, Abhishek Agarwal, Manoj Sarda, Vatsal Sanjay, Arup Kumar Das
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Fire accidents in trains bring huge disaster to human life and property. Evacuation becomes a major challenge in such incidents owing to confined spaces, large passenger density and trains moving at high speeds. The pantry car in Indian Railways trains carry inflammable materials like cooking fuel and LPG and electrical fittings. The pantry car is therefore highly susceptible to fire accidents. Numerical simulations have been done in a pantry car of Indian locomotive train using computational fluid dynamics based software. Different scenarios of a fire outbreak have been explored by varying Heat Release Rate per Unit Area (HRRPUA) of the fire source, introduction of exhaust in the cooking area, and taking a case of an air conditioned pantry car. Temporal statures of flame and soot have been obtained for each scenario and differences have been studied and reported. Inputs from this study can be used to assess casualties in fire accidents in locomotive trains and development of smoke control/detection systems in Indian trains.Keywords: fire propagation, flame contour, pantry fire, soot flow
Procedia PDF Downloads 3391312 Development of Wound Dressing System Based on Hydrogel Matrix Incorporated with pH-Sensitive Nanocarrier-Drug Systems
Authors: Dagmara Malina, Katarzyna Bialik-Wąs, Klaudia Pluta
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The growing significance of transdermal systems, in which skin is a route for systemic drug delivery, has generated a considerable amount of data which has resulted in a deeper understanding of the mechanisms of transport across the skin in the context of the controlled and prolonged release of active substances. One of such solutions may be the use of carrier systems based on intelligent polymers with different physicochemical properties. In these systems, active substances, e.g. drugs, can be conjugated (attached), immobilized, or encapsulated in a polymer matrix that is sensitive to specific environmental conditions (e.g. pH or temperature changes). Intelligent polymers can be divided according to their sensitivity to specific environmental stimuli such as temperature, pH, light, electric, magnetic, sound, or electromagnetic fields. Materials & methods—The first stage of the presented research concerned the synthesis of pH-sensitive polymeric carriers by a radical polymerization reaction. Then, the selected active substance (hydrocortisone) was introduced into polymeric carriers. In a further stage, bio-hybrid sodium alginate/poly(vinyl alcohol) – SA/PVA-based hydrogel matrices modified with various carrier-drug systems were prepared with the chemical cross-linking method. The conducted research included the assessment of physicochemical properties of obtained materials i.e. degree of hydrogel swelling and degradation studies as a function of pH in distilled water and phosphate-buffered saline (PBS) at 37°C in time. The gel fraction represents the insoluble gel fraction as a result of inter-molecule cross-linking formation was also measured. Additionally, the chemical structure of obtained hydrogels was confirmed using FT-IR spectroscopic technique. The dynamic light scattering (DLS) technique was used for the analysis of the average particle size of polymer-carriers and carrier-drug systems. The nanocarriers morphology was observed using SEM microscopy. Results & Discussion—The analysis of the encapsulated polymeric carriers showed that it was possible to obtain the time-stable empty pH-sensitive carrier with an average size 479 nm and the encapsulated system containing hydrocortisone with an average 543 nm, which was introduced into hydrogel structure. Bio-hybrid hydrogel matrices are stable materials, and the presence of an additional component: pH-sensitive carrier – hydrocortisone system, does not reduce the degree of cross-linking of the matrix nor its swelling ability. Moreover, the results of swelling tests indicate that systems containing higher concentrations of the drug have a slightly higher sorption capacity in each of the media used. All analyzed materials show stable and statically changing swelling values in simulated body fluids - there is no sudden fluid uptake and no rapid release from the material. The analysis of FT-IR spectra confirms the chemical structure of the obtained bio-hybrid hydrogel matrices. In the case of modifications with a pH-sensitive carrier, a much more intense band can be observed in the 3200-3500 cm⁻¹ range, which most likely originates from the strong hydrogen interactions that occur between individual components.Keywords: hydrogels, polymer nanocarriers, sodium alginate/poly(vinyl alcohol) matrices, wound dressings.
Procedia PDF Downloads 1461311 Blood Lipid Management: Combined Treatment with Hydrotherapy and Ozone Bubbles Bursting in Water
Authors: M. M. Wickramasinghe
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Cholesterol and triglycerides are lipids, mainly essential to maintain the cellular structure of the human body. Cholesterol is also important for hormone production, vitamin D production, proper digestion functions, and strengthening the immune system. Excess fats in the blood circulation, known as hyperlipidemia, become harmful leading to arterial clogging and causing atherosclerosis. Aim of this research is to develop a treatment protocol to efficiently break down and maintain circulatory lipids by improving blood circulation without strenuous physical exercises while immersed in a tub of water. To achieve the target of strong exercise effect, this method involves generating powerful ozone bubbles to spin, collide, and burst in the water. Powerful emission of air into water is capable of transferring locked energy of the water molecules and releasing energy. This method involves water and air-based impact generated by pumping ozone at the speed of 46 lts/sec with a concentration of 0.03-0.05 ppt according to safety standards of The Federal Institute for Drugs and Medical Devices, BfArM, Germany. The direct impact of ozone bubbles on the muscular system and skin becomes the main target and is capable of increasing the heart rate while immersed in water. A total time duration of 20 minutes is adequate to exert a strong exercise effect, improve blood circulation, and stimulate the nervous and endocrine systems. Unstable ozone breakdown into oxygen release onto the surface of the water giving additional benefits and supplying high-quality air rich in oxygen required to maintain efficient metabolic functions. The breathing technique was introduced to improve the efficiency of lung functions and benefit the air exchange mechanism. The temperature of the water is maintained at 39c to 40c to support arterial dilation and enzyme functions and efficiently improve blood circulation to the vital organs. The buoyancy of water and natural hydrostatic pressure release the tension of the body weight and relax the mind and body. Sufficient hydration (3lts of water per day) is an essential requirement to transport nutrients and remove waste byproducts to process through the liver, kidney, and skin. Proper nutritional intake is an added advantage to optimize the efficiency of this method which aids in a fast recovery process. Within 20-30 days of daily treatment, triglycerides, low-density lipoproteins (LDL), and total cholesterol reduction were observed in patients with abnormal levels of lipid profile. Borderline patients were cleared within 10–15 days of treatment. This is a highly efficient system that provides many benefits and is able to achieve a successful reduction of triglycerides, LDL, and total cholesterol within a short period of time. Supported by proper hydration and nutritional balance, this system of natural treatment maintains healthy levels of lipids in the blood and avoids the risk of cerebral stroke, high blood pressure, and heart attacks.Keywords: atherosclerosis, cholesterol, hydrotherapy, hyperlipidemia, lipid management, ozone therapy, triglycerides
Procedia PDF Downloads 911310 Prediction of Ionizing Radiation Doses in Irradiated red Pepper (Capsicum annuum) and Mint (Mentha piperita) by Gel Electrophoresis
Authors: Şeyma Özçirak Ergün, Ergün Şakalar, Emrah Yalazi̇, Nebahat Şahi̇n
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Food irradiation is a usage of exposing food to ionising radiation (IR) such as gamma rays. IR has been used to decrease the number of harmful microorganisms in the food such as spices. Excessive usage of IR can cause damage to both food and people who consuming food. And also it causes to damages on food DNA. Generally, IR detection techniques were utilized in literature for spices are Electron Spin Resonance (ESR), Thermos Luminescence (TL). Storage creates negative effect on IR detection method then analyses of samples have been performed without storage in general. In the experimental part, red pepper (Capsicum annuum) and mint (Mentha piperita) as spices were exposed to 0, 0.272, 0.497, 1.06, 3.64, 8.82, and 17.42 kGy ionize radiation. ESR was applied to samples irradiated. DNA isolation from irradiated samples was performed using GIDAGEN Multi Fast DNA isolation kit. The DNA concentration was measured using a microplate reader spectrophotometer (Infinite® 200 PRO-Life Science–Tecan). The concentration of each DNA was adjusted to 50 ng/µL. Genomic DNA was imaged by UV transilluminator (Gel Doc XR System, Bio-Rad) for the estimation of genomic DNA bp-fragment size after IR. Thus, agarose gel profiles of irradiated spices were obtained to determine the change of band profiles. Besides, samples were examined at three different time periods (0, 3, 6 months storage) to show the feasibility of developed method. Results of gel electrophoresis showed especially degradation of DNA of irradiated samples. In conclusion, this study with gel electrophoresis can be used as a basis for the identification of the dose of irradiation by looking at degradation profiles at specific amounts of irradiation. Agarose gel results of irradiated samples were confirmed with ESR analysis. This method can be applied widely to not only food products but also all biological materials containing DNA to predict radiation-induced damage of DNA.Keywords: DNA, electrophoresis, gel electrophoresis, ionizeradiation
Procedia PDF Downloads 2591309 HLA-G, a Neglected Immunosuppressive Checkpoint for Breast Cancer Immunotherapy
Authors: Xian-Peng Jiang, Catherine C. Baucom, Toby Jiang, Robert L. Elliott
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HLA-G binds to the inhibitory receptors of uterine NK cells and plays an important role in protection of fetal cells from maternal NK lysis. HLA-G also mediates tumor escape, but the immunosuppressive role is often neglected. These studies have focused on the examination of HLA-G expression in human breast carcinoma and HLA-G immunosuppressive role in NK cytolysis. We examined HLA-G expression in breast cell lines by real time PCR, ELISA and immunofluorescent staining. We treated the breast cancer cell lines with anti-human HLA-G antibody or progesterone. Then, NK cytolysis was measured by using MTT assay. We find that breast carcinoma cell lines increase the expression of HLA-G mRNA and protein, compared to normal cells. Blocking HLA-G of the breast cancer cells by the antibody increases NK cytolysis. Progesterone upregulates HLA-G mRNA and protein of human breast cancer cell lines. The increased HLA-G expression suppresses NK cytolysis. In summary, human breast carcinoma overexpress HLA-G immunosuppressive molecules. Blocking HLA-G protein by antibody improves NK cytolysis. In contrast, upregulation of HLA-G expression by progesterone impairs NK cytolytic function. Thus, HLA-G is a new immunosuppressive checkpoint and potential cancer immunotherapeutic target.Keywords: HLA-G, Breast carcinoma, NK cells, Immunosuppressive checkpoint
Procedia PDF Downloads 881308 Assessment of Genotoxic Effects of a Fungicide (Propiconazole) in Freshwater Fish Gambusia Affinis Using Alkaline Single-Cell Gel Electrophoresis (Comet Essay)
Authors: Bourenane Bouhafs Naziha
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ARTEA330EC is a fungicide used to inhibit the growth of many types of fungi on and cereals and rice, it is the single largest selling agrochemical that has been widely detected in surface waters in our area (Northeast Algerian). The studies on long-term genotoxic effects of fugicides in different tissues of fish using genotoxic biomarkers are limited. Therefore, in the present study DNA damage by propiconazole in freshwater fish Gambusia affinis by comet assays was investigated. The LC(50)- 96 h of the fungicide was estimated for the fish in a semi-static system. On this basis of LC(50) value sublethal and nonlethal concentrations were determined (25; 50; 75; and 100 ppm). The DNA damage was measured in erythrocytes as the percentage of DNA in comet tails of fishes exposed to above concentrations the fungicide. In general,non significant effects for both the concentrations and time of exposure were observed in treated fish compared with the controls. However It was found that the highest DNA damage was observed at the highest concentration and the longest time of exposure (day 12). The study indicated comet assay to be sensitive and rapid method to detect genotoxicity of propiconasol and other pesticides in fishes.Keywords: genotoxicity, fungicide, propiconazole, freshwater, Gambusia affinis, alkaline single-cell gel electrophoresis
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