Search results for: oxygen reduction reaction (ORR)
5620 Prevalence and Molecular Characterization of Vibrio parahaemolyticus in Estuarine Fish from Dhaka City Markets
Authors: Fahmida Khalique Nitu
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Little is known on the biosafety level of Vibrio parahaemolyticus in estuarine fish in Bangladesh. The purpose of this study was to investigate the prevalence and concentration of V. parahaemolyticus in estuarine fishes using the Polymerase Chain Reaction( PCR) method . The study was conducted on 37 fishes of different species from different types of estuarine fish commonly sold at city markets. Sampling was done on the intestinal tract and gills of each fish. The prevalence of V. parahaemolyticus was found to be 29.72% with higher percentages detected in samples from the gills (89.28%) followed by the intestinal tract (10.71%). The density of Vibrio spp. in the gill of estuarine fishes with an average was 4.4 x103CFU/g and in the intestine samples was 1.5x103 CFU/g. The outcome of the biosafety assessment V. parahaemolyticus in estuarine fish indicates another potential source of food safety issues to consumers.Keywords: biosafety, estuarine, prevalence, Vibrios
Procedia PDF Downloads 2825619 Biocompatible Hydrogel Materials Containing Cytostatics for Cancer Treatment
Authors: S. Kudlacik-Kramarczyk, M. Kedzierska, B. Tyliszczak
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Recently, the continuous development of medicine and related sciences has been observed. Particular emphasis is directed on the development of biomaterials, i.e., non-toxic, biocompatible and biodegradable materials that may improve the effectiveness of treatment as well as the comfort of patients. This is particularly important in the case of cancer treatment. Currently, there are many methods of cancer treatment based primarily on chemotherapy and the surgical removal of the tumor, but it is worth noting that these therapies also cause many side effects. Among women, the most common cancer is breast cancer. It may be completely cured, but the consequence of treatment is partial or complete breast mastectomy and radiation therapy, which results in severe skin burns. The skin of the patient after radiation therapy is very burned, and therefore requires intensive care and high frequency of dressing changes. The traditional dressing adheres to the burn wounds and does not absorb adequate amount of exudate from injuries and the patient is forced to change the dressing every 2 hours. Therefore, the main purpose was to develop an innovative combination of dressing material with drug carriers that may be used in anti-cancer therapy. The innovation of this solution is the combination of these two products into one system, i.e., a transdermal system with the possibility of a controlled release of the drug- cytostatic. Besides, the possibility of modifying the hydrogel matrix with aloe vera juice provides this material with new features favorable from the point of view of healing processes of burn wounds resulting from the radiation therapy. In this study, hydrogel materials containing protein spheres with the active substance have been obtained as a result of photopolymerization process. The reaction mixture consisting of the protein (albumin) spheres incorporated with cytostatic, chitosan, adequate crosslinking agent and photoinitiator has been subjected to the UV radiation for 2 minutes. Prepared materials have been subjected to the numerous studies including the analysis of cytotoxicity using murine fibroblasts L929. Analysis was conducted based on the mitochondrial activity test (MTT reduction assay) which involves the determining the number of cells characterized by proper metabolism. Hydrogel materials obtained using different amount of crosslinking agents have been subjected to the cytotoxicity analysis. According to the standards, tested material is defined as cytotoxic when the viability of cells after 24 h incubation with this material is lower than 70%. In the research, hydrogel polymer materials containing protein spheres incorporated with the active substance, i.e. a cytostatic, have been developed. Such a dressing may support the treatment of cancer due to the content of the anti-cancer drug - cytostatic, and may also provide a soothing effect on the healing of the burn wounds resulted from the radiation therapy due to the content of aloe vera juice in the hydrogel matrix. Based on the conducted cytotoxicity studies, it may be concluded that the obtained materials do not adversely affect the tested cell lines, therefore they can be subjected to more advanced analyzes.Keywords: hydrogel polymers, cytostatics, drug carriers, cytotoxicity
Procedia PDF Downloads 1345618 Monitoring CO2 and H2S Emission in Live Austrian and UK Concrete Sewer Pipes
Authors: Anna Romanova, Morteza A. Alani
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Corrosion of concrete sewer pipes induced by sulfuric acid is an acknowledged problem and a ticking time-bomb to sewer operators. Whilst the chemical reaction of the corrosion process is well-understood, the indirect roles of other parameters in the corrosion process which are found in sewer environment are not highly reflected on. This paper reports on a field studies undertaken in Austria and United Kingdom, where the parameters of temperature, pH, H2S and CO2 were monitored over a period of time. The study establishes that (i) effluent temperature and pH have similar daily pattern and peak times, When examined in minutes scale, (ii) H2S and CO2 have an identical hourly pattern, (iii) H2S instant or shifted relation to effluent temperature is governed by the root mean square value of CO2.Keywords: concrete corrosion, carbon dioxide, hydrogen sulphide, sewer pipe, sulfuric acid
Procedia PDF Downloads 3115617 Examining the Impact of Degrees of Slag Replacement on the Carbonation Process of Slag-Blended Cement
Authors: Geta Bekalu Belayneh, Solmoi Park
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This study examines the role of slag in the process of hydration and carbonation of carbonation-cured slag cement. Carbonation-cured slag-blended cement paste samples were prepared with varying slag percentages of 0%, 10%, 30%, and 50%. The curing process lasted for a maximum of 28 days. The findings demonstrated that the carbonation depth increased as the curing period was extended, and a larger slag percentage promoted a more extensive penetration of carbonation. The degree of belite reaction was greatly enhanced in the slag-blended cement, resulting in an increased ability to bind CO₂ in the blended cement. These findings enhance comprehension of the behaviour of blended cement produced through carbonation-curing, facilitating the advancement of more environmentally friendly and long-lasting concrete constructions.Keywords: carbonation curing, blast furnace slag, characterization, Portland cement
Procedia PDF Downloads 745616 Biological Activity of Hibiscus sabdariffa Extract
Authors: Chanasit Chaocharoenphat
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Hibiscus sabdariffa is a herbal plant that is commonly used for home remedies in Thailand. This study aims to determine the antioxidant activity of polyphenols, as oxidative stress plays a vital role in the development of cancer, and H. sabdariffa was used in this study. The total flavonoids content was determined using the aluminium chloride colourimetric method and expressed as quercetin equivalents (QE)/g and the antioxidant capacity of the flavonoids using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging capacity assays. The IC50 values of H. sabdariffa extract were 167.14 μg/mL ± 0.843 and 77.59 μg/mL ± 0.798, respectively. In the DPPH assay, vitamin C was used as a positive control, whereas Trolox was used as a positive control in the ABTS assay. To summarise, H. sabdariffa extract contains a high concentration of total flavonoids and exhibits potent antioxidant activity. However, additional antioxidant activity assays such as superoxide dismutase (SOD), reactive oxygen species (ROS), and reactive nitrogen species (RNS) scavenging assays and in vitro antioxidant experiments should be carried out to investigate the molecular mechanism of the compound.Keywords: ABTS assay, antioxidant activity, Gracilaria fisheri, DPPH assays, total flavonoid content
Procedia PDF Downloads 2555615 Hypoxia Tolerance, Longevity and Cancer-Resistance in the Mole Rat Spalax – a Liver Transcriptomics Approach
Authors: Hanno Schmidt, Assaf Malik, Anne Bicker, Gesa Poetzsch, Aaron Avivi, Imad Shams, Thomas Hankeln
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The blind subterranean mole rat Spalax shows a remarkable tolerance to hypoxia, cancer-resistance and longevity. Unravelling the genomic basis of these adaptations will be important for biomedical applications. RNA-Seq gene expression data were obtained from normoxic and hypoxic Spalax and rat liver tissue. Hypoxic Spalax broadly downregulates genes from major liver function pathways. This energy-saving response is likely a crucial adaptation to low oxygen levels. In contrast, the hypoxiasensitive rat shows massive upregulation of energy metabolism genes. Candidate genes with plausible connections to the mole rat’s phenotype, such as important key genes related to hypoxia-tolerance, DNA damage repair, tumourigenesis and ageing, are substantially higher expressed in Spalax than in rat. Comparative liver transcriptomics highlights the importance of molecular adaptations at the gene regulatory level in Spalax and pinpoints a variety of starting points for subsequent functional studies.Keywords: cancer, hypoxia, longevity, transcriptomics
Procedia PDF Downloads 1625614 Effects of Sulphide Mining on AISI 304 Stainless Steel
Authors: Aguasanta Miguel Sarmiento, José Miguel Dávila, María Luisa de la Torre
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Acid mine drainage (AMD) is an acidic leachate with high levels of metals and sulphates in solution, which seriously affects the durability and strength of metallic materials used in the construction of structural and mechanical components. This paper presents the results of the evolution over time of the reduction in tensile strength and defects in AISI 304 stainless steel in contact with acid mine drainage. For this purpose, a total of 30 bars with a diameter of 8 mm and a length of 14 cm were placed transversely in the course of a stream contaminated by AMD from the sulphide mines of the Iberian Pyritic Belt (SW Spain). This stream has average pH values of 2.6, a potential of 660 mV, and average concentrations of 12 g/L of sulphates, 1.2 g/L of Fe, 191 mg/L of Zn, etc. Every two months of exposure, 6 stainless steel bars were extracted from the acid stream. They were subjected to surface roughness analysis carried out with the help of Mitutoyo Surftest SJ-210 surface roughness tester. The analysis was carried out at three different points on 5 specimens from each series. The average reading of each parameter is calculated in order to ensure the accuracy of the measurements and the surface coverage. Arithmetic mean roughness value (Ra), mean roughness depth (Rz), and root mean square roughness (Rq) were measured. Five specimens from each series were statically tensile tested using universal equipment (Servosis ME 403 of 200kN). The specimens were clamped at their ends with two grips for cylindrical sections, and the tensile force was applied at a constant speed of 0.5 kN/s, according to the requirements of standard UNE-EN ISO 6892-1: 2020. To determine the modulus of elasticity, limits close to 15% and 55% of the maximum load were used, depending on the course of each test. Field Emission Scanning Electron Microscopy (FESEM) was used to observe corrosion products and defects generated by exposure to AMD. Energy dispersive X-ray spectrometry (EDS) was used to analyse the chemical composition of the corrosion products formed. For this purpose, small pieces were cut from the resulting specimens, cleaned, and embedded in epoxy resin. The results show that after only 5 months of exposure of AISI 304 stainless steel to the mining environment, the surface roughness increases significantly, with average depths almost 6 times greater than the initial one. Cracks are observed on the surface of the material, which increases in size with the time of exposure. A large number of grains with a composition of more than 57% Pb and 16% Sn can be observed inside these cracks. Tensile tests show a reduction in the resistance of this material after only two months of exposure. The results show the serious problems that would result from the use of this material for the use of mechanical components in a sulphide mining environment, not only because of the significant reduction in the lifetime of such components, but also because of the implications for human safety.Keywords: acid mine drainage, corrosion, mechanical properties, stainless steel
Procedia PDF Downloads 255613 Granulomatous Mycoses Fungoides: A Case Report
Authors: Girum Tedla Assefa
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Background: Granulomatous mycosis fungoides is an extremely rare type of cutaneous T-cell lymphoma (<55 cases reported worldwide). Case report: A 36-year-old female presented with soft tissue atrophy of right lower limb (dermis + hypodermis) of 22 years and plaques over trunk of 3 years duration. Histological examination of a biopsy taken from the atrophied tissue showed a granulomatous reaction with epidermotropic atypical lymphocytes. However, in other areas there were only findings of conventional MF without granuloma. Conclusion: The diagnosis of a granulomatous mycosis fungoides depends exclusively on the histological demonstration of granulomas. Distinct clinical characteristics are not present. This case highlights the importance of thorough investigation of lipoatrophic skin changes in the adult to exclude underlying causes, including MF.Keywords: cutaneous lymphoma, granulomatous skin lymphoma, mycoses fungoides, skin atrophy
Procedia PDF Downloads 3735612 Development of 111In-DOTMP as a New Bone Imaging Agent
Authors: H. Yousefnia, S. Zolghadri, AR. Jalilian, A. Mirzaei, A. Bahrami-Samani, M. Erfani
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The objective of this study is the preparation of 111In-DOTMP as a new bone imaging agent. 111In was produced at the Agricultural, Medical and Industrial Research School (AMIRS) by means of 30 MeV cyclotron via natCd(p,x)111In reaction. Complexion of In‐111 with DOTMP was carried out by adding 0.1 ml of the stock solution (50 mg/ml in 2 N NaoH) to the vial containing 1 mCi of 111In. pH of the mixture was adjusted to 7-8 by means of phosphate buffer. The radiochemical purity of the complex at the optimized condition was higher than 98% (by using whatman No.1 paper in NH4OH:MeOH: H2O (0.2:2:4)). Both the biodistribution studies and SPECT imaging indicated high bone uptake. The ratio of bone to other soft tissue accumulation was significantly high which permit to observe high quality images. The results show that 111In-DOTMP can be used as a suitable tracer for diagnosis of bone metastases by SPECT imaging.Keywords: biodistribution, DOTMP, 111In, SPECT
Procedia PDF Downloads 5345611 Genetic Characterization of Barley Genotypes via Inter-Simple Sequence Repeat
Authors: Mustafa Yorgancılar, Emine Atalay, Necdet Akgün, Ali Topal
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In this study, polymerase chain reaction based Inter-simple sequence repeat (ISSR) from DNA fingerprinting techniques were used to investigate the genetic relationships among barley crossbreed genotypes in Turkey. It is important that selection based on the genetic base in breeding programs via ISSR, in terms of breeding time. 14 ISSR primers generated a total of 97 bands, of which 81 (83.35%) were polymorphic. The highest total resolution power (RP) value was obtained from the F2 (0.53) and M16 (0.51) primers. According to the ISSR result, the genetic similarity index changed between 0.64–095; Lane 3 with Line 6 genotypes were the closest, while Line 36 were the most distant ones. The ISSR markers were found to be promising for assessing genetic diversity in barley crossbreed genotypes.Keywords: barley, crossbreed, genetic similarity, ISSR
Procedia PDF Downloads 3525610 Leaching of Flotation Concentrate of Oxide Copper Ore from Sepon Mine, Lao PDR
Authors: C. Rattanakawin, S. Vasailor
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Acid leaching of flotation concentrate of oxide copper ore containing mainly of malachite was performed in a standard agitation tank with various parameters. The effects of solid to liquid ratio, sulfuric acid concentration, agitation speed, leaching temperature and time were examined to get proper conditions. The best conditions are 1:8 solid to liquid ratio, 10% concentration by weight, 250 rev/min, 30 oC and 5-min leaching time in respect. About 20% Cu grade assayed by atomic absorption technique with 98% copper recovery was obtained from these combined optimum conditions. Dissolution kinetics of the concentrate was approximated as a logarithmic function. As a result, the first-order reaction rate is suggested from this leaching study.Keywords: agitation leaching, dissolution kinetics, flotation concentrate, oxide copper ore, sulfuric acid
Procedia PDF Downloads 1245609 Characteristic on Compressive Strength of Blast Slag and Fly Ash Hybrid Geopolymer Mortar
Authors: G. S. Ryu, K. T. Koh, H. Y. Kim, G. H. An, D. W. Seo
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Geopolymer mortar is produced by alkaline activation of pozzolanic materials such as fly ground granulated blast-furnace slag (GGBFS) and fly ash (FA). Its unique reaction pathway facilitates rapid strength development in comparison with hydration of ordinary Portland cement (OPC). Geopolymer can be fabricated using various types and dosages of alkali-activator, which effectively gives a wider control over the performance of the final product. The present study investigates the effect of types of precursors and curing conditions on the fresh state and strength development characteristics of geopolymers, thereby comparatively exploring the effect of precursors from various sources of origin. The obtained result showed that the setting time and strength development of the specimens with the identical mix proportion but different precursors displayed significant variations.Keywords: alkali-activated material, blast furnace slag, fly ash, flowability, strength development
Procedia PDF Downloads 2525608 Biosynthesis of a Nanoparticle-Antibody Phthalocyanine Photosensitizer for Use in Targeted Photodynamic Therapy of Cervical Cancer
Authors: Elvin P. Chizenga, Heidi Abrahamse
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Cancer cell resistance to therapy is the main cause of treatment failures and the poor prognosis of cancer convalescence. The progression of cervical cancer to other parts of the genitourinary system and the reported recurrence rates are overwhelming. Current treatments, including surgery, chemo and radiation have been inefficient in eradicating the tumor cells. These treatments are also associated with poor prognosis and reduced quality of life, including fertility loss. This has inspired the need for the development of new treatment modalities to eradicate cervical cancer successfully. Photodynamic Therapy (PDT) is a modern treatment modality that induces cell death by photochemical interactions of light and a photosensitizer, which in the presence of molecular oxygen, yields a set of chemical reactions that generate Reactive Oxygen Species (ROS) and other free radical species causing cell damage. Enhancing PDT using modified drug delivery can increase the concentration of the photosensitizer in the tumor cells, and this has the potential to maximize its therapeutic efficacy. In cervical cancer, all infected cells constitutively express genes of the E6 and E7 HPV viral oncoproteins, resulting in high concentrations of E6 and E7 in the cytoplasm. This provides an opportunity for active targeting of cervical cancer cells using immune-mediated drug delivery to maximize therapeutic efficacy. The use of nanoparticles in PDT has also proven effective in enhancing therapeutic efficacy. Gold nanoparticles (AuNps) in particular, are explored for their use in biomedicine due to their biocompatibility, low toxicity, and enhancement of drug uptake by tumor cells. In this present study, a biomolecule comprising of AuNPs, anti-E6 monoclonal antibodies, and Aluminium Phthalocyanine photosensitizer was synthesized for use in targeted PDT of cervical cancer. The AuNp-Anti-E6-Sulfonated Aluminium Phthalocyanine mix (AlPcSmix) photosensitizing biomolecule was synthesized by coupling AuNps and anti-E6 monoclonal antibodies to the AlPcSmix via Polyethylene Glycol (PEG) chemical links. The final product was characterized using Transmission Electron Microscope (TEM), Zeta Potential, Uv-Vis Spectrophotometry, Fourier Transform Infrared Spectroscopy (FTIR), and X-ray diffraction (XRD), to confirm its chemical structure and functionality. To observe its therapeutic role in treating cervical cancer, cervical cancer cells, HeLa cells were seeded in 3.4 cm² diameter culture dishes at a concentration of 5x10⁵ cells/ml, in vitro. The cells were treated with varying concentrations of the photosensitizing biomolecule and irradiated using a 673.2 nm wavelength of laser light. Post irradiation cellular responses were performed to observe changes in morphology, viability, proliferation, cytotoxicity, and cell death pathways induced. Dose-Dependent response of the cells to treatment was demonstrated as significant morphologic changes, increased cytotoxicity, and decreased cell viability and proliferation This study presented a synthetic biomolecule for targeted PDT of cervical cancer. The study suggested that PDT using this AuNp- Anti-E6- AlPcSmix photosensitizing biomolecule is a very effective treatment method for the eradication of cervical cancer cells, in vitro. Further studies in vivo need to be conducted to support the use of this biomolecule in treating cervical cancer in clinical settings.Keywords: anti-E6 monoclonal antibody, cervical cancer, gold nanoparticles, photodynamic therapy
Procedia PDF Downloads 1285607 PLGA Nanoparticles Entrapping dual anti-TB drugs of Amikacin and Moxifloxacin as a Potential Host-Directed Therapy for Multidrug Resistant Tuberculosis
Authors: Sharif Abdelghany
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Polymeric nanoparticles have been widely investigated as a controlled release drug delivery platform for the treatment of tuberculosis (TB). These nanoparticles were also readily internalised into macrophages, leading to high intracellular drug concentration. In this study two anti-TB drugs, amikacin and moxifloxacin were encapsulated into PLGA nanoparticles. The novelty of this work appears in: (1) the efficient encapsulation of two hydrophilic second-line anti-TB drugs, and (2) intramacrophage delivery of this synergistic combination potentially for rapid treatment of multi-drug resistant TB (MDR-TB). Two water-oil-water (w/o/w) emulsion strategies were employed in this study: (1) alginate coated PLGA nanoparticles, and (2) alginate entrapped PLGA nanoparticles. The average particle size and polydispersity index (PDI) of the alginate coated PLGA nanoparticles were found to be unfavourably high with values of 640 ± 32 nm and 0.63 ± 0.09, respectively. In contrast, the alginate entrapped PLGA nanoparticles were within the desirable particle size range of 282 - 315 nm and the PDI was 0.08 - 0.16, and therefore were chosen for subsequent studies. Alginate entrapped PLGA nanoparticles yielded a drug loading of over 10 µg/mg powder for amikacin, and more than 5 µg/mg for moxifloxacin and entrapment efficiencies range of approximately 25-31% for moxifloxacin and 51-59% for amikacin. To study macrophage uptake efficiency, the nanoparticles of alginate entrapped nanoparticle formulation were loaded with acridine orange as a marker, seeded to THP-1 derived macrophages and viewed under confocal microscopy. The particles were readily internalised into the macrophages and highly concentrated in the nucleus region. Furthermore, the anti-mycobacterial activity of the drug-loaded particles was evaluated using M. tuberculosis-infected macrophages, which revealed a significant reduction (4 log reduction) of viable bacterial count compared to the untreated group. In conclusion, the amikacin-moxifloxacin alginate entrapped PLGA nanoparticles are promising for further in vivo studies.Keywords: moxifloxacin and amikacin, nanoparticles, multidrug resistant TB, PLGA
Procedia PDF Downloads 3705606 Combined Treatment of Aged Rats with Donepezil and the Gingko Extract EGb 761® Enhances Learning and Memory Superiorly to Monotherapy
Authors: Linda Blümel, Bettina Bert, Jan Brosda, Heidrun Fink, Melanie Hamann
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Age-related cognitive decline can eventually lead to dementia, the most common mental illness in elderly people and an immense challenge for patients, their families and caregivers. Cholinesterase inhibitors constitute the most commonly used antidementia prescription medication. The standardized Ginkgo biloba leaf extract EGb 761® is approved for treating age-associated cognitive impairment and has been shown to improve the quality of life in patients suffering from mild dementia. A clinical trial with 96 Alzheimer´s disease patients indicated that the combined treatment with donepezil and EGb 761® had fewer side effects than donepezil alone. In an animal model of cognitive aging, we compared the effect of combined treatment with EGb 761® or donepezil monotherapy and vehicle. We compared the effect of chronic treatment (15 days of pretreatment) with donepezil (1.5 mg/kg p. o.), EGb 761® (100 mg/kg p. o.), or the combination of the two drugs, or vehicle in 18 – 20 month old male OFA rats. Learning and memory performance were assessed by Morris water maze testing, motor behavior in an open field paradigm. In addition to chronic treatment, the substances were administered orally 30 minutes before testing. Compared to the first day and to the control group, only the combination group showed a significant reduction in latency to reach the hidden platform on the second day of testing. Moreover, from the second day of testing onwards, the donepezil, the EGb 761® and the combination group required less time to reach the hidden platform compared to the first day. The control group did not reach the same latency reduction until day three. There were no effects on motor behavior. These results suggest a superiority of the combined treatment of donepezil with EGb 761® compared to monotherapy.Keywords: age-related cognitive decline, dementia, ginkgo biloba leaf extract EGb 761®, learning and memory, old rats
Procedia PDF Downloads 3705605 Development of a Thermodynamic Model for Ladle Metallurgy Steel Making Processes Using Factsage and Its Macro Facility
Authors: Prasenjit Singha, Ajay Kumar Shukla
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To produce high-quality steel in larger volumes, dynamic control of composition and temperature throughout the process is essential. In this paper, we developed a mass transfer model based on thermodynamics to simulate the ladle metallurgy steel-making process using FactSage and its macro facility. The overall heat and mass transfer processes consist of one equilibrium chamber, two non-equilibrium chambers, and one adiabatic reactor. The flow of material, as well as heat transfer, occurs across four interconnected unit chambers and a reactor. We used the macro programming facility of FactSage™ software to understand the thermochemical model of the secondary steel making process. In our model, we varied the oxygen content during the process and studied their effect on the composition of the final hot metal and slag. The model has been validated with respect to the plant data for the steel composition, which is similar to the ladle metallurgy steel-making process in the industry. The resulting composition profile serves as a guiding tool to optimize the process of ladle metallurgy in steel-making industries.Keywords: desulphurization, degassing, factsage, reactor
Procedia PDF Downloads 2235604 Efficacy and Safety of Sublingual Sufentanil for the Management of Acute Pain
Authors: Neil Singla, Derek Muse, Karen DiDonato, Pamela Palmer
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Introduction: Pain is the most common reason people visit emergency rooms. Studies indicate however, that Emergency Department (ED) physicians often do not provide adequate analgesia to their patients as a result of gender and age bias, opiophobia and insufficient knowledge of and formal training in acute pain management. Novel classes of analgesics have recently been introduced, but many patients suffer from acute pain in settings where the availability of intravenous (IV) access may be limited, so there remains a clinical need for rapid-acting, potent analgesics that do not require an invasive route of delivery. A sublingual sufentanil tablet (SST), dispensed using a single-dose applicator, is in development for treatment of moderate-to-severe acute pain in a medically-supervised setting. Objective: The primary objective of this study was to demonstrate the repeat-dose efficacy, safety and tolerability of sufentanil 20 mcg and 30 mcg sublingual tablets compared to placebo for the management of acute pain as determined by the time-weighted sum of pain intensity differences (SPID) to baseline over the 12-hour study period (SPID12). Key secondary efficacy variables included SPID over the first hour (SPID1), Total pain relief over the 12-hour study period (TOTPAR12), time to perceived pain relief (PR) and time to meaningful PR. Safety variables consisted of adverse events (AE), vital signs, oxygen saturation and early termination. Methods: In this Phase 2, double-blind, dose-finding study, an equal number of male and female patients were randomly assigned in a 2:2:1 ratio to SST 20 mcg, SS 30 mcg or placebo, respectively, following bunionectomy. Study drug was dosed as needed, but not more frequently than hourly. Rescue medication was available as needed. The primary endpoint was the Summed Pain Intensity Difference to baseline over 12h (SPIDI2). Safety was assessed by continuous oxygen saturation monitoring and adverse event reporting. Results: 101 patients (51 Male/50 Female) were randomized, 100 received study treatment (intent-to-treat [ITT] population), and 91 completed the study. Reasons for early discontinuation were lack of efficacy (6), adverse events (2) and drug-dosing error (1). Mean age was 42.5 years. For the ITT population, SST 30 mcg was superior to placebo (p=0.003) for the SPID12. SPID12 scores in the active groups were superior for both male (ANOVA overall p-value =0.038) and female (ANOVA overall p-value=0.005) patients. Statistically significant differences in favour of sublingual sufentanil were also observed between the SST 30mcg and placebo group for SPID1(p<0.001), TOTPAR12(p=0.002), time to perceived PR (p=0.023) and time to meaningful PR (p=0.010). Nausea, vomiting and somnolence were more frequent in the sufentanil groups but there were no significant differences between treatment arms for the proportion of patients who prematurely terminated due to AE or inadequate analgesia. Conclusions: Sufentanil tablets dispensed sublingually using a single-dose applicator is in development for treatment of patients with moderate-to-severe acute pain in a medically-supervised setting where immediate IV access is limited. When administered sublingually, sufentanil’s pharmacokinetic profile and non-invasive delivery makes it a useful alternative to IM or IV dosing.Keywords: acute pain, pain management, sublingual, sufentanil
Procedia PDF Downloads 3595603 Fresh Amnion Membrane Grafting for the Regeneration of Skin in Full Thickness Burn in Newborn - Case Report
Authors: Priyanka Yadav, Umesh Bnasal, Yashvinder Kumar
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The placenta is an important structure that provides oxygen and nutrients to the growing fetus in utero. It is usually thrown away after birth, but it has a therapeutic role in the regeneration of tissue. It is covered by the amniotic membrane, which can be easily separated into the amnion layer and the chorion layer—the amnion layer act as a biofilm for the healing of burn wound and non-healing ulcers. The freshly collected membrane has stem cells, cytokines, growth factors, and anti-inflammatory properties, which act as a biofilm for the healing of wounds. It functions as a barrier and prevents heat and water loss and also protects from bacterial contamination, thus supporting the healing process. The application of Amnion membranes has been successfully used for wound and reconstructive purposes for decades. It is a very cheap and easy process and has shown superior results to allograft and xenograft. However, there are very few case reports of amnion membrane grafting in newborns; we intend to highlight its therapeutic importance in burn injuries in newborns. We present a case of 9 days old male neonate who presented to the neonatal unit of Maulana Azad Medical College with a complaint of fluid-filled blisters and burns wound on the body for six days. He was born outside the hospital at 38 weeks of gestation to a 24-year-old primigravida mother by vaginal delivery. The presentation was cephalic and the amniotic fluid was clear. His birth weight was 2800 gm and APGAR scores were 7 and 8 at 1 and 5 minutes, respectively. His anthropometry was appropriate for gestational age. He developed respiratory distress after birth requiring oxygen support by nasal prongs for three days. On the day of life three, he developed blisters on his body, starting from than face then over the back and perineal region. At a presentation on the day of life nine, he had blisters and necrotic wound on the right side of the face, back, right shoulder and genitalia, affecting 60% of body surface area with full-thickness loss of skin. He was started on intravenous antibiotics and fluid therapy. Pus culture grew Pseudomonas aeuroginosa, for which culture-specific antibiotics were started. Plastic surgery reference was taken and regular wound dressing was done with antiseptics. He had a storming course during the hospital stay. On the day of life 35 when the baby was hemodynamically stable, amnion membrane grafting was done on the wound site; for the grafting, fresh amnion membrane was removed under sterile conditions from the placenta obtained by caesarean section. It was then transported to the plastic surgery unit in half an hour in a sterile fluid where the graft was applied over the infant’s wound. The amnion membrane grafting was done twice in two weeks for covering the whole wound area. After successful uptake of amnion membrane, skin from the thigh region was autografted over the whole wound area by Meek technique in a single setting. The uptake of autograft was excellent and most of the areas were healed. In some areas, there was patchy regeneration of skin so dressing was continued. The infant was discharged after three months of hospital stay and was later followed up in the plastic surgery unit of the hospital.Keywords: amnion membrane grafting, autograft, meek technique, newborn, regeneration of skin
Procedia PDF Downloads 1645602 Comparison of Chemical Coagulation and Electrocoagulation for Boron Removal from Synthetic Wastewater Using Aluminium
Authors: Kartikaningsih Danis, Yao-Hui Huang
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Various techniques including conventional and advanced have been employed for the boron treatment from water and wastewater. The electrocoagulation involves an electrolytic reactor for coagulation/flotation with aluminium as anode and cathode. There is aluminium as coagulant to be used for removal which may induce secondary pollution in chemical coagulation. The purpose of this study is to investigate and compare the performance between electrocoagulation and chemical coagulation on boron removal from synthetic wastewater. The effect of different parameters, such as pH reaction, coagulant dosage, and initial boron concentration were examined. The results show that the boron removal using chemical coagulation was lower. At the optimum condition (e.g. pH 8 and 0.8 mol coagulant dosage), boron removal efficiencies for chemical coagulation and electrocoagulation were 61% and 91%, respectively. In addition, the electrocoagulation needs no chemical reagents and makes the boron treatment easy for application.Keywords: boron removal, chemical coagulation, aluminum, electro-coagulation
Procedia PDF Downloads 4075601 Evaluation of the Cytotoxicity and Genotoxicity of Chemical Material in Filters PM2.5 of the Monitoring Stations of the Network of Air Quality in the Valle De Aburrá, Colombia
Authors: Alejandra Betancur Sánchez, Carmen Elena Zapata Sánchez, Juan Bautista López Ortiz
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Adverse effects and increased air pollution has raised concerns about regulatory policies and has fostered the development of new air quality standards; this is due to the complexity of the composition and the poorly understood reactions in the atmospheric environment. Toxic compounds act as environmental agents having various effects, from irritation to death of cells and tissues. A toxic agent is defined an adverse response in a biological system. There is a particular class that produces some kind of alteration in the genetic material or associated components, so they are recognized as genotoxic agents. Within cells, they interact directly or indirectly with DNA, causing mutations or interfere with some enzymatic repair processes or in the genesis or polymerization of proteinaceous material involved in chromosome segregation. An air pollutant may cause or contribute to increased mortality or serious illness and even pose a potential danger to human health. The aim of this study was to evaluate the effect on the viability and the genotoxic potential on the cell lines CHO-K1 and Jurkat and peripheral blood of particulate matter PM T lymphocytes 2.5 obtained from filters collected three monitoring stations network air quality Aburrá Valley. Tests, reduction of MTT, trypan blue, NRU, comet assay, sister chromatid exchange (SCE) and chromosomal aberrations allowed evidence reduction in cell viability in cell lines CHO-K1 and Jurkat and damage to the DNA from cell line CHOK1, however, no significant effects were observed in the number of SCEs and chromosomal aberrations. The results suggest that PM2.5 material has genotoxic potential and can induce cancer development, as has been suggested in other studies.Keywords: PM2.5, cell line Jurkat, cell line CHO-K1, cytotoxicity, genotoxicity
Procedia PDF Downloads 2675600 Coal Fly Ash Based Ceramic Membrane for Water Purification via Ultrafiltration
Authors: Obsi Terfasa, Bhanupriya Das, Shiao-Shing Chen
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Converting coal fly ash (CFA) waste into ceramic membranes presents a promising alternative to traditional disposal methods, offering potential economic and environmental advantages that warrant further investigation. This research focuses on the creation of ceramic membranes exclusively from CFA using a uniaxial compaction technique. The membranes' properties were examined through various analytical methods: Scanning Electron Microscopy (SEM) revealed a porous and flawless membrane surface, X-Ray Diffraction (XRD) identified mullite and quartz crystalline structures, and Fourier-Transform Infrared Spectroscopy (FTIR) characterized the membrane's functional groups. Thermogravimetric analysis (TGA) determined the ideal sintering temperature to be 800°C. To evaluate its separation capabilities, the synthesized membrane was tested on wastewater from denim jeans production at 0.2 bar pressure. The results were impressive, with 97.42% removal of Chemical Oxygen Demand (COD), 95% color elimination, and a pure water flux of 4.5 Lm⁻²h⁻¹bar⁻¹. These findings suggest that CFA, a byproduct of thermal power plants, can be effectively repurposed to produce ultrafiltration membranes suitable for various industrial purification and separations.Keywords: wastewater treatment, separator, coal fly ash, ceramic membrane, ultrafiltration
Procedia PDF Downloads 435599 Superparamagnetic Core Shell Catalysts for the Environmental Production of Fuels from Renewable Lignin
Authors: Cristina Opris, Bogdan Cojocaru, Madalina Tudorache, Simona M. Coman, Vasile I. Parvulescu, Camelia Bala, Bahir Duraki, Jeroen A. Van Bokhoven
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The tremendous achievements in the development of the society concretized by more sophisticated materials and systems are merely based on non-renewable resources. Consequently, after more than two centuries of intensive development, among others, we are faced with the decrease of the fossil fuel reserves, an increased impact of the greenhouse gases on the environment, and economic effects caused by the fluctuations in oil and mineral resource prices. The use of biomass may solve part of these problems, and recent analyses demonstrated that from the perspective of the reduction of the emissions of carbon dioxide, its valorization may bring important advantages conditioned by the usage of genetic modified fast growing trees or wastes, as primary sources. In this context, the abundance and complex structure of lignin may offer various possibilities of exploitation. However, its transformation in fuels or chemicals supposes a complex chemistry involving the cleavage of C-O and C-C bonds and altering of the functional groups. Chemistry offered various solutions in this sense. However, despite the intense work, there are still many drawbacks limiting the industrial application. Thus, the proposed technologies considered mainly homogeneous catalysts meaning expensive noble metals based systems that are hard to be recovered at the end of the reaction. Also, the reactions were carried out in organic solvents that are not acceptable today from the environmental point of view. To avoid these problems, the concept of this work was to investigate the synthesis of superparamagnetic core shell catalysts for the fragmentation of lignin directly in the aqueous phase. The magnetic nanoparticles were covered with a nanoshell of an oxide (niobia) with a double role: to protect the magnetic nanoparticles and to generate a proper (acidic) catalytic function and, on this composite, cobalt nanoparticles were deposed in order to catalyze the C-C bond splitting. With this purpose, we developed a protocol to prepare multifunctional and magnetic separable nano-composite Co@Nb2O5@Fe3O4 catalysts. We have also established an analytic protocol for the identification and quantification of the fragments resulted from lignin depolymerization in both liquid and solid phase. The fragmentation of various lignins occurred on the prepared materials in high yields and with very good selectivity in the desired fragments. The optimization of the catalyst composition indicated a cobalt loading of 4wt% as optimal. Working at 180 oC and 10 atm H2 this catalyst allowed a conversion of lignin up to 60% leading to a mixture containing over 96% in C20-C28 and C29-C37 fragments that were then completely fragmented to C12-C16 in a second stage. The investigated catalysts were completely recyclable, and no leaching of the elements included in the composition was determined by inductively coupled plasma optical emission spectrometry (ICP-OES).Keywords: superparamagnetic core-shell catalysts, environmental production of fuels, renewable lignin, recyclable catalysts
Procedia PDF Downloads 3335598 A One-Dimensional Modeling Analysis of the Influence of Swirl and Tumble Coefficient in a Single-Cylinder Research Engine
Authors: Mateus Silva Mendonça, Wender Pereira de Oliveira, Gabriel Heleno de Paula Araújo, Hiago Tenório Teixeira Santana Rocha, Augusto César Teixeira Malaquias, José Guilherme Coelho Baeta
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The stricter legislation and the greater demand of the population regard to gas emissions and their effects on the environment as well as on human health make the automotive industry reinforce research focused on reducing levels of contamination. This reduction can be achieved through the implementation of improvements in internal combustion engines in such a way that they promote the reduction of both specific fuel consumption and air pollutant emissions. These improvements can be obtained through numerical simulation, which is a technique that works together with experimental tests. The aim of this paper is to build, with support of the GT-Suite software, a one-dimensional model of a single-cylinder research engine to analyze the impact of the variation of swirl and tumble coefficients on the performance and on the air pollutant emissions of an engine. Initially, the discharge coefficient is calculated through the software Converge CFD 3D, given that it is an input parameter in GT-Power. Mesh sensitivity tests are made in 3D geometry built for this purpose, using the mass flow rate in the valve as a reference. In the one-dimensional simulation is adopted the non-predictive combustion model called Three Pressure Analysis (TPA) is, and then data such as mass trapped in cylinder, heat release rate, and accumulated released energy are calculated, aiming that the validation can be performed by comparing these data with those obtained experimentally. Finally, the swirl and tumble coefficients are introduced in their corresponding objects so that their influences can be observed when compared to the results obtained previously.Keywords: 1D simulation, single-cylinder research engine, swirl coefficient, three pressure analysis, tumble coefficient
Procedia PDF Downloads 1085597 Synthesis of Ni/Mesopore Silica-Alumina Catalyst for Hydrocracking of Pyrolyzed α-Cellulose
Authors: Wega Trisunaryanti, Hesty Kusumastuti, Iip Izul Falah, Muhammad Fajar Marsuki, Rahmad Nuryanto
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Synthesis of Ni supported on mesopore silica-alumina (MSA) for hydrocracking of pyrolyzed α-cellulose had been carried out. The silica and alumina were extracted from Sidoarjo mud. Gelatin from catfish bone was used as a template for the mesopore design. The MSA was synthesized by using hydrothermal method at 100 °C for 24 h and calcined at 550 °C for 4 h then characterized by using X-Ray Diffraction Spectrometer (XRD) and Nitrogen Gas Sorption Analyzer (GAS). The Ni metal was loaded to the MSA by wet impregnation method. The catalytic activity in the hydrocracking reaction of pyrolyzed α-cellulose was carried out at 450 °C for 2 h. The MSA synthesized in this work is an amorphous material with specific surface area, total pore volume, and average pore diameter of 212.29 m²/g, 1.29 cm³/g, and 20.05 nm, respectively. The Ni/MSA catalyst produced 73.02 wt.% of liquid product in hydrocracking of pyrolyzed α-cellulose.Keywords: catalyst, gelatin, hydrocracking, mesopore silica-alumina, α-cellulose
Procedia PDF Downloads 1655596 Mobility of Metallic Trace Elements (MTE) in Water and Sediment of the Rivers: Case of Nil River, North-Eastern Algerian
Authors: S. Benessam, T. H. Debieche, S. Amiour, A. Chine, S. Khelili
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The metallic trace elements (MTE) are present in water and sediments of the rivers with weak concentrations. Several physicochemical parameters (Eh, pH and oxygen dissolved) and chemical processes (adsorption, absorption, complexation and precipitation) as well as nature of the sediments control their mobility. In order to determine the effect of these factors on the mobility of some MTE (Cd, Cr, Cu, Fe, Pb and Zn) in water of the rivers, a two-monthly monitoring of the physicochemical parameters and chemistry of water and sediments of the Nil wadi (Algeria) was carried out during the period from November 2013 to January 2015. The results show that each MTE has its own conditions of mobility and generally are very influence by the variations of the pH and Eh. Under the natural conditions, neutral pH with basic and medium oxidizing, only the lead presented in water with raised values, indicating its solubility in water and its salting out of the sediments. The other MTE present raised concentrations in the sediments, indicating their trapping by adsorption and/or chemical precipitation. The chemical form of each ETM was given by Eh-pH diagrams. The spatio-temporal monitoring of these ETM shows the effect of the rains, the dry periods and the rejects in the variation of their concentrations.Keywords: chemistry, metallic trace elements, sediment, water
Procedia PDF Downloads 2945595 Challenges in Achieving Profitability for MRO Companies in the Aviation Industry: An Analytical Approach
Authors: Nur Sahver Uslu, Ali̇ Hakan Büyüklü
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Maintenance, Repair, and Overhaul (MRO) costs are significant in the aviation industry. On the other hand, companies that provide MRO services to the aviation industry but are not dominant in the sector, need to determine the right strategies for sustainable profitability in a competitive environment. This study examined the operational real data of a small medium enterprise (SME) MRO company where analytical methods are not widely applied. The company's customers were divided into two categories: airline companies and non-airline companies, and the variables that best explained profitability were analyzed with Logistic Regression for each category and the results were compared. First, data reduction was applied to the transformed variables that went through the data cleaning and preparation stages, and the variables to be included in the model were decided. The misclassification rates for the logistic regression results concerning both customer categories are similar, indicating consistent model performance across different segments. Less profit margin is obtained from airline customers, which can be explained by the variables part description, time to quotation (TTQ), turnaround time (TAT), manager, part cost, and labour cost. The higher profit margin obtained from non-airline customers is explained only by the variables part description, part cost, and labour cost. Based on the two models, it can be stated that it is significantly more challenging for the MRO company, which is the subject of our study, to achieve profitability from Airline customers. While operational processes and organizational structure also affect the profit from airline customers, only the type of parts and costs determine the profit for non-airlines.Keywords: aircraft, aircraft components, aviation, data analytics, data science, gini index, maintenance, repair, and overhaul, MRO, logistic regression, profit, variable clustering, variable reduction
Procedia PDF Downloads 415594 Fabrication and Characterization of Glass Nanofibers through Electrospinning of Silica Sol-Gel along with in situ Synthesis of Ag Nanoparticles
Authors: Mahsa Kangazian Kangazi, Ali Akbar Ghareh Aghaji, Majid Montazer
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Nowadays, silica nanofibers are highly regarded among the inorganic nanofibers due to the high reactivity and availability of silicon compounds in nature. Sol-gel process is required for electrospinning of silica nanofibers in which a metal alkoxide is hydrolyzed, and the viscosity is increased. In this study, silica nanofibers containing silver nanoparticles were synthesized and electrospun from a mixture of silica sol with an easy spinnable polymer (PVA) as an additive. The silica sol contains tetraethyl orthosilicate (TEOS), silver nitrate, distilled water, nitric acid, and ethanol. Nanofibers were formed through electrospinning setup. The nanofibers were calcinated to remove the solvent and additive polymer. Consequently, pure silica nanofibers were produced. FTIR analysis indicated entire removal of polyvinyl alcohol from the structure and formation of silan groups. The presence of silver, silica and oxygen was confirmed by EDX. Also, XRD patterns revealed the presence of silver nanoparticles with a mean crystal size of 18 nm. FESEM images showed that adding silver nitrate into the sol-gel, resulted in lower nanofibers diameter from 286 to 136 nm. Furthermore, the electrospun nanofibers were more resistance in acidic media than alkaline media.Keywords: in situ synthesis of silver nanoparticles, silica nanofibers, sol-gel, tetraethyl orthosilicate
Procedia PDF Downloads 1845593 Stainless Steel Degradation by Sulphide Mining
Authors: Aguasanta M. Sarmiento, Jose Miguel Davila, Juan Carlos Fortes, Maria Luisa de la Torre
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Acid mine drainage (AMD) is an acidic leachate with high levels of metals and sulphates in solution, which seriously affects the durability and strength of metallic materials used in the construction of structural and mechanical components. This paper presents the results of the evolution over time of the reduction in tensile strength and defects in AISI 304 stainless steel in contact with acid mine drainage. For this purpose, a total of 30 bars with a diameter of 8 mm and a length of 14 cm were placed transversely in the course of a stream contaminated by AMD from the sulphide mines of the Iberian Pyritic Belt (SW Spain). This stream has average pH values of 2.6, a potential of 660 mV and average concentrations of 12 g/L of sulphates, 1.2 g/L of Fe, 191 mg/L of Zn, etc. Every two months of exposure, 6 stainless steel bars were extracted from the acid stream. They were subjected to surface roughness analysis carried out with the help of Mitutoyo Surftest SJ-210 surface roughness tester. The analysis was carried out at three different points on 5 specimens from each series. The average reading of each parameter is calculated in order to ensure the accuracy of the measurements and the surface coverage. Arithmetic mean roughness value (Ra), mean roughness depth (Rz) and root mean square roughness (Rq) were measured. Five specimens from each series were statically tensile tested using universal equipment (Servosis ME 403 of 200kN). The specimens were clamped at their ends with two grips for cylindrical sections, and the tensile force was applied at a constant speed of 0.5 kN/s, according to the requirements of standard UNE-EN ISO 6892-1: 2020. To determine the modulus of elasticity, limits close to 15% and 55% of the maximum load were used, depending on the course of each test. Field Emission Scanning Electron Microscopy (FESEM) was used to observe corrosion products and defects generated by exposure to AMD. Energy dispersive X-ray spectrometry (EDS) was used to analyze the chemical composition of the corrosion products formed. For this purpose, small pieces were cut from the resulting specimens, cleaned and embedded in epoxy resin. The results show that after only 5 months of exposure of AISI 304 stainless steel to the mining environment, the surface roughness increases significantly, with average depths almost 6 times greater than the initial one. Cracks are observed on the surface of the material, which increases in size with the time of exposure. A large number of grains with a composition of more than 57% Pb and 16% Sn can be observed inside these cracks. Tensile tests show a reduction in the resistance of this material after only two months of exposure. The results show the serious problems that would result from the use of this material for the use of mechanical components in a sulphide mining environment, not only because of the significant reduction in the lifetime of such components but also because of the implications for human safety.Keywords: Acid mine drainage, Corrosion, Mechanical properties, Stainless steel
Procedia PDF Downloads 165592 Polarization of Glass with Positive and Negative Charge Carriers
Authors: Valentina V. Zhurikhina, Mihail I. Petrov, Alexandra A. Rtischeva, Mark Dussauze, Thierry Cardinal, Andrey A. Lipovskii
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Polarization of glass, often referred to as thermal poling, is a well-known method to modify the glass physical and chemical properties, that manifest themselves in loosing central symmetry of the medium, glass structure and refractive index modification. The usage of the poling for second optical harmonic generation, fabrication of optical waveguides and electrooptic modulators was also reported. Nevertheless, the detailed description of the poling of glasses, containing multiple charge carriers is still under discussion. In particular, the role of possible migration of electrons in the space charge formation usually remains out of the question. In this work, we performed the numerical simulation of thermal poling of a silicate glass, containing Na, K, Mg, and Ca. We took into consideration the contribution of electrons in the polarization process. The possible explanation of migration of electrons can be the break of non-bridging oxygen bonds. It was found, that the modeled depth of the space charge region is about 10 times higher if the migration of the negative charges is taken under consideration. The simulated profiles of cations, participating in the polarization process, are in a good agreement with the experimental data, obtained by glow discharge spectroscopy.Keywords: glass poling, charge transport, modeling, concentration profiles
Procedia PDF Downloads 3615591 The Molecular Mechanism of Vacuolar Function in Yeast Cell Homeostasis
Authors: Chang-Hui Shen, Paulina Konarzewska
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Cell homeostasis is regulated by vacuolar activity and it has been shown that lipid composition of the vacuole plays an important role in vacuolar function. The major phosphoinositide species present in the vacuolar membrane include phosphatidylinositol 3,5-biphosphate (PI(3,5)P₂) which is generated from PI(3)P controlled by Fab1p. Deletion of FAB1 gene reduce the synthesis of PI(3,5)P₂ and thus result in enlarged or fragmented vacuoles, with neutral vacuolar pH due to reduced vacuolar H⁺-ATPase activity. These mutants also exhibited poor growth at high extracellular pH and in the presence of CaCl₂. Conversely, VPS34 regulates the synthesis of PI(3)P from phosphatidylinositol (PI), and the lack of Vps34p results in the reduction of vacuolar activity. Although the cellular observations are clear, it is still unknown about the molecular mechanism between the phospholipid biosynthesis pathway and vacuolar activity. Since both VPS34 and FAB1 are important in vacuolar activity, we hypothesize that the molecular mechanism of vacuolar function might be regulated by the transcriptional regulators of phospholipid biosynthesis. In this study, we study the role of the major phospholipid biosynthesis transcription factor, INO2, in the regulation of vacuolar activity. We first performed qRT-PCR to examine the effect of Ino2p on the expression of VPS34 and FAB1. Our results showed that VPS34 was upregulated in the presence of inositol for both WT and ino2Δ cells. However, FAB1 was only upregulated significantly in ino2Δ cells. This indicated that Ino2p might be the negative regulator for FAB1 expression. Next, growth sensitivity experiment showed that WT, vma3Δ, and ino2Δ grew well in growth medium buffered to pH 5.5 containing 10 mM CaCl₂. As cells were switched to growth medium buffered to pH 7 containing CaCl₂ WT, ino2Δ and opi1Δ showed growth reduction, whereas vma3Δ was completely nonviable. As the concentration of CaCl₂ was increased to 60 mM, ino2Δ cells showed moderate growth reduction compared to WT. This result suggests that ino2Δ cells have better vacuolar activity. Microscopic analysis and vacuolar acidification were employed to further elucidate the importance of INO2 in vacuolar homeostasis. Analysis of vacuolar morphology indicated that WT and vma3Δ cells displayed vacuoles that occupied a small area of the cell when grown in media buffered to pH 5.5. Whereas, ino2Δ displayed fragmented vacuoles. On the other hand, all strains grown in media buffered to pH 7, exhibited enlarged vacuoles that occupied most of the cell’s surface. This indicated that the presence of INO2 may play negative effect in vacuolar morphology when cells are grown in media buffered to pH 5.5. Furthermore, vacuolar acidification assay showed that only vma3Δ cells displayed notably less acidic vacuoles as cells were grown in media buffered to pH 5.5 and pH 7. Whereas, ino2Δ cells displayed more acidic pH compared to WT at pH7. Taken together, our results demonstrated the molecular mechanism of the vacuolar activity regulated by the phospholipid biosynthesis transcription factors Ino2p. Ino2p negatively regulates vacuolar activity through the expression of FAB1.Keywords: vacuole, phospholipid, homeostasis, Ino2p, FAB1
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