Search results for: injection
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 908

Search results for: injection

98 Retrospective Analysis of 142 Cases of Incision Infection Complicated with Sternal Osteomyelitis after Cardiac Surgery Treated by Activated PRP Gel Filling

Authors: Daifeng Hao, Guang Feng, Jingfeng Zhao, Tao Li, Xiaoye Tuo

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Objective: To retrospectively analyze the clinical characteristics of incision infection with sternal osteomyelitis sinus tract after cardiac surgery and the operation method and therapeutic effect of filling and repairing with activated PRP gel. Methods: From March 2011 to October 2022, 142 cases of incision infection after cardiac surgery with sternal osteomyelitis sinus were retrospectively analyzed, and the causes of poor wound healing after surgery, wound characteristics, perioperative wound management were summarized. Treatment during operation, collection and storage process of autologous PRP before debridement surgery, PRP filling repair and activation method after debridement surgery, effect of anticoagulant drugs on surgery, postoperative complications and average wound healing time, etc.. Results: Among the cases in this group, 53.3% underwent coronary artery bypass grafting, 36.8% underwent artificial heart valve replacement, 8.2% underwent aortic artificial vessel replacement, and 1.7% underwent allogeneic heart transplantation. The main causes of poor incision healing were suture reaction, fat liquefaction, osteoporosis, diabetes, and metal allergy in sequence. The wound is characterized by an infected sinus tract. Before the operation, 100-150ml of PRP with 4 times the physiological concentration was collected separately with a blood component separation device. After sinus debridement, PRP was perfused to fill the bony defect in the middle of the sternum, activated with thrombin freeze-dried powder and calcium gluconate injection to form a gel, and the outer skin and subcutaneous tissue were sutured freely. 62.9% of patients discontinued warfarin during the perioperative period, and 37.1% of patients maintained warfarin treatment. There was no significant difference in the incidence of postoperative wound hematoma. The average postoperative wound healing time was 12.9±4.7 days, and there was no obvious postoperative complication. Conclusions: Application of activated PRP gel to fill incision infection with sternal osteomyelitis sinus after cardiac surgery has a less surgical injury and satisfactory and stable curative effect. It can completely replace the previously used pectoralis major muscle flap transplantation operation scheme.

Keywords: platelet-rich plasma, negative-pressure wound therapy, sternal osteomyelitis, cardiac surgery

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97 Giant Cancer Cell Formation: A Link between Cell Survival and Morphological Changes in Cancer Cells

Authors: Rostyslav Horbay, Nick Korolis, Vahid Anvari, Rostyslav Stoika

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Introduction: Giant cancer cells (GCC) are common in all types of cancer, especially after poor therapy. Some specific features of such cells include ~10-fold enlargement, drug resistance, and the ability to propagate similar daughter cells. We used murine NK/Ly lymphoma, an aggressive and fast growing lymphoma model that has already shown drastic changes in GCC comparing to parental cells (chromatin condensation, nuclear fragmentation, tighter OXPHOS/cellular respiration coupling, multidrug resistance). Materials and methods: In this study, we compared morpho-functional changes of GCC that predominantly show either a cytostatic or a cytotoxic effect after treatment with drugs. We studied the effect of a combined cytostatic/cytotoxic drug treatment to determine the correlation of drug efficiency and GCC formation. Doses of G1/S-specific drug paclitaxel/PTX (G2/M-specific, 50 mg/mouse), vinblastine/VBL (50 mg/mouse), and DNA-targeting agents doxorubicin/DOX (125 ng/mouse) and cisplatin/CP (225 ng/mouse) on C57 black mice. Several tests were chosen to estimate morphological and physiological state (propidium iodide, Rhodamine-123, DAPI, JC-1, Janus Green, Giemsa staining and other), which included cell integrity, nuclear fragmentation and chromatin condensation, mitochondrial activity, and others. A single and double factor ANOVA analysis were performed to determine correlation between the criteria of applied drugs and cytomorphological changes. Results: In all cases of treatment, several morphological changes were observed (intracellular vacuolization, membrane blebbing, and interconnected mitochondrial network). A lower gain in ascites (49.97% comparing to control group) and longest lifespan (22+9 days) after tumor injection was obtained with single VBL and single DOX injections. Such ascites contained the highest number of GCC (83.7%+9.2%), lowest cell count number (72.7+31.0 mln/ml), and a strong correlation coefficient between increased mitochondrial activity and percentage of giant NK/Ly cells. A high number of viable GCC (82.1+9.2%) was observed compared to the parental forms (15.4+11.9%) indicating that GCC are more drug resistant than the parental cells. All this indicates that the giant cell formation and its ability to obtain drug resistance is an expanding field in cancer research.

Keywords: ANOVA, cisplatin, doxorubicin, drug resistance, giant cancer cells, NK/Ly lymphoma, paclitaxel, vinblastine

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96 Radiomics: Approach to Enable Early Diagnosis of Non-Specific Breast Nodules in Contrast-Enhanced Magnetic Resonance Imaging

Authors: N. D'Amico, E. Grossi, B. Colombo, F. Rigiroli, M. Buscema, D. Fazzini, G. Cornalba, S. Papa

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Purpose: To characterize, through a radiomic approach, the nature of nodules considered non-specific by expert radiologists, recognized in magnetic resonance mammography (MRm) with T1-weighted (T1w) sequences with paramagnetic contrast. Material and Methods: 47 cases out of 1200 undergoing MRm, in which the MRm assessment gave uncertain classification (non-specific nodules), were admitted to the study. The clinical outcome of the non-specific nodules was later found through follow-up or further exams (biopsy), finding 35 benign and 12 malignant. All MR Images were acquired at 1.5T, a first basal T1w sequence and then four T1w acquisitions after the paramagnetic contrast injection. After a manual segmentation of the lesions, done by a radiologist, and the extraction of 150 radiomic features (30 features per 5 subsequent times) a machine learning (ML) approach was used. An evolutionary algorithm (TWIST system based on KNN algorithm) was used to subdivide the dataset into training and validation test and to select features yielding the maximal amount of information. After this pre-processing, different machine learning systems were applied to develop a predictive model based on a training-testing crossover procedure. 10 cases with a benign nodule (follow-up older than 5 years) and 18 with an evident malignant tumor (clear malignant histological exam) were added to the dataset in order to allow the ML system to better learn from data. Results: NaiveBayes algorithm working on 79 features selected by a TWIST system, resulted to be the best performing ML system with a sensitivity of 96% and a specificity of 78% and a global accuracy of 87% (average values of two training-testing procedures ab-ba). The results showed that in the subset of 47 non-specific nodules, the algorithm predicted the outcome of 45 nodules which an expert radiologist could not identify. Conclusion: In this pilot study we identified a radiomic approach allowing ML systems to perform well in the diagnosis of a non-specific nodule at MR mammography. This algorithm could be a great support for the early diagnosis of malignant breast tumor, in the event the radiologist is not able to identify the kind of lesion and reduces the necessity for long follow-up. Clinical Relevance: This machine learning algorithm could be essential to support the radiologist in early diagnosis of non-specific nodules, in order to avoid strenuous follow-up and painful biopsy for the patient.

Keywords: breast, machine learning, MRI, radiomics

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95 Energy Efficiency Measures in Canada’s Iron and Steel Industry

Authors: A. Talaei, M. Ahiduzzaman, A. Kumar

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In Canada, an increase in the production of iron and steel is anticipated for satisfying the increasing demand of iron and steel in the oil sands and automobile industries. It is predicted that GHG emissions from iron and steel sector will show a continuous increase till 2030 and, with emissions of 20 million tonnes of carbon dioxide equivalent, the sector will account for more than 2% of total national GHG emissions, or 12% of industrial emissions (i.e. 25% increase from 2010 levels). Therefore, there is an urgent need to improve the energy intensity and to implement energy efficiency measures in the industry to reduce the GHG footprint. This paper analyzes the current energy consumption in the Canadian iron and steel industries and identifies energy efficiency opportunities to improve the energy intensity and mitigate greenhouse gas emissions from this industry. In order to do this, a demand tree is developed representing different iron and steel production routs and the technologies within each rout. The main energy consumer within the industry is found to be flared heaters accounting for 81% of overall energy consumption followed by motor system and steam generation each accounting for 7% of total energy consumption. Eighteen different energy efficiency measures are identified which will help the efficiency improvement in various subsector of the industry. In the sintering process, heat recovery from coolers provides a high potential for energy saving and can be integrated in both new and existing plants. Coke dry quenching (CDQ) has the same advantages. Within the blast furnace iron-making process, injection of large amounts of coal in the furnace appears to be more effective than any other option in this category. In addition, because coal-powered electricity is being phased out in Ontario (where the majority of iron and steel plants are located) there will be surplus coal that could be used in iron and steel plants. In the steel-making processes, the recovery of Basic Oxygen Furnace (BOF) gas and scrap preheating provides considerable potential for energy savings in BOF and Electric Arc Furnace (EAF) steel-making processes, respectively. However, despite the energy savings potential, the BOF gas recovery is not applicable in existing plants using steam recovery processes. Given that the share of EAF in steel production is expected to increase the application potential of the technology will be limited. On the other hand, the long lifetime of the technology and the expected capacity increase of EAF makes scrap preheating a justified energy saving option. This paper would present the results of the assessment of the above mentioned options in terms of the costs and GHG mitigation potential.

Keywords: Iron and Steel Sectors, Energy Efficiency Improvement, Blast Furnace Iron-making Process, GHG Mitigation

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94 Strategies of Drug Discovery in Insects

Authors: Alaaeddeen M. Seufi

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Many have been published on therapeutic derivatives from living organisms including insects. In addition to traditional maggot therapy, more than 900 therapeutic products were isolated from insects. Most people look at insects as enemies and others believe that insects are friends. Many beneficial insects rather than Honey Bees, Silk Worms and Shellac insect could insure human-insect friendship. In addition, insects could be MicroFactories, Biosensors or Bioreactors. InsectFarm is an amazing example of the applied research that transfers insects from laboratory to market by Prof Mircea Ciuhrii and co-workers. They worked for 18 years to derive therapeutics from insects. Their research resulted in production of more than 30 commercial medications derived from insects (e.g. Imunomax, Noblesse, etc.). Two general approaches were followed to discover drugs from living organisms. Some laboratories preferred biochemical approach to purify components of the innate immune system of insects and insect metabolites as well. Then the purified components could be tested for many therapeutic trials. Other researchers preferred molecular approach based on proteomic studies. Components of the innate immune system of insects were then tested for their medical activities. Our Laboratory team preferred to induce insect immune system (using oral, topical and injection routes of administration), then a transcriptomic study was done to discover the induced genes and to identify specific biomarkers that can help in drug discovery. Biomarkers play an important role in medicine and in drug discovery and development as well. Optimum biomarker development and application will require a team approach because of the multifaceted nature of biomarker selection, validation, and application. This team uses several techniques such as pharmacoepidemiology, pharmacogenomics, and functional proteomics; bioanalytical development and validation; modeling and simulation to improve and refine drug development. Our Achievements included the discovery of four components of the innate immune system of Spodoptera littoralis and Musca domestica. These components were designated as SpliDef (defesin), SpliLec (lectin), SpliCec (cecropin) and MdAtt (attacin). SpliDef, SpliLec and MdAtt were confirmed as antimicrobial peptides, while SpliCec was additionally confirmed as anticancer peptide. Our current research is going on to achieve something in antioxidants and anticoagulants from insects. Our perspective is to achieve something in the mass production of prototypes of our products and to reach it to the commercial level. These achievements are the integrated contributions of everybody in our team staff.

Keywords: AMPs, insect, innate immunitty, therappeutics

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93 Antimicrobial Properties of SEBS Compounds with Copper Microparticles

Authors: Vanda Ferreira Ribeiro, Daiane Tomacheski, Douglas Naue Simões, Michele Pitto, Ruth Marlene Campomanes Santana

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Indoor environments, such as car cabins and public transportation vehicles are places where users are subject to air quality. Microorganisms (bacteria, fungi, yeasts) enter these environments through windows, ventilation systems and may use the organic particles present as a growth substrate. In addition, atmospheric pollutants can act as potential carbon and nitrogen sources for some microorganisms. Compounds base SEBS copolymers, poly(styrene-b-(ethylene-co-butylene)-b-styrene, are a class of thermoplastic elastomers (TPEs), fully recyclable and largely used in automotive parts. Metals, such as cooper and silver, have biocidal activities and the production of the SEBS compounds by melting blending with these agents can be a good option for producing compounds for use in plastic parts of ventilation systems and automotive air-conditioning, in order to minimize the problems caused by growth of pathogenic microorganisms. In this sense, the aim of this work was to evaluate the effect of copper microparticles as antimicrobial agent in compositions based on SEBS/PP/oil/calcite. Copper microparticles were used in weight proportion of 0%, 1%, 2% and 4%. The compounds were prepared using a co-rotating double screw extruder (L/D ratio of 40/1 and 16 mm screw diameter). The processing parameters were 300 rpm of screw rotation rate, with a temperature profile between 150 to 190°C. SEBS based TPE compounds were injection molded. The compounds emission were characterized by gravimetric fogging test. Compounds were characterized by physical (density and staining by contact), mechanical (hardness and tension properties) and rheological properties (melt volume rate – MVR). Antibacterial properties were evaluated against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) strains. To avaluate the abilities toward the fungi have been chosen Aspergillus niger (A. niger), Candida albicans (C. albicans), Cladosporium cladosporioides (C. cladosporioides) and Penicillium chrysogenum (P. chrysogenum). The results of biological tests showed a reduction on bacteria in up to 88% in E.coli and up to 93% in S. aureus. The tests with fungi showed no conclusive results because the sample without copper also demonstrated inhibition of the development of these microorganisms. The copper addition did not cause significant variations in mechanical properties, in the MVR and the emission behavior of the compounds. The density increases with the increment of copper in compounds.

Keywords: air conditioner, antimicrobial, cooper, SEBS

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92 Stability Indicating RP – HPLC Method Development, Validation and Kinetic Study for Amiloride Hydrochloride and Furosemide in Pharmaceutical Dosage Form

Authors: Jignasha Derasari, Patel Krishna M, Modi Jignasa G.

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Chemical stability of pharmaceutical molecules is a matter of great concern as it affects the safety and efficacy of the drug product.Stability testing data provides the basis to understand how the quality of a drug substance and drug product changes with time under the influence of various environmental factors. Besides this, it also helps in selecting proper formulation and package as well as providing proper storage conditions and shelf life, which is essential for regulatory documentation. The ICH guideline states that stress testing is intended to identify the likely degradation products which further help in determination of the intrinsic stability of the molecule and establishing degradation pathways, and to validate the stability indicating procedures. A simple, accurate and precise stability indicating RP- HPLC method was developed and validated for simultaneous estimation of Amiloride Hydrochloride and Furosemide in tablet dosage form. Separation was achieved on an Phenomenexluna ODS C18 (250 mm × 4.6 mm i.d., 5 µm particle size) by using a mobile phase consisting of Ortho phosphoric acid: Acetonitrile (50:50 %v/v) at a flow rate of 1.0 ml/min (pH 3.5 adjusted with 0.1 % TEA in Water) isocratic pump mode, Injection volume 20 µl and wavelength of detection was kept at 283 nm. Retention time for Amiloride Hydrochloride and Furosemide was 1.810 min and 4.269 min respectively. Linearity of the proposed method was obtained in the range of 40-60 µg/ml and 320-480 µg/ml and Correlation coefficient was 0.999 and 0.998 for Amiloride hydrochloride and Furosemide, respectively. Forced degradation study was carried out on combined dosage form with various stress conditions like hydrolysis (acid and base hydrolysis), oxidative and thermal conditions as per ICH guideline Q2 (R1). The RP- HPLC method has shown an adequate separation for Amiloride hydrochloride and Furosemide from its degradation products. Proposed method was validated as per ICH guidelines for specificity, linearity, accuracy; precision and robustness for estimation of Amiloride hydrochloride and Furosemide in commercially available tablet dosage form and results were found to be satisfactory and significant. The developed and validated stability indicating RP-HPLC method can be used successfully for marketed formulations. Forced degradation studies help in generating degradants in much shorter span of time, mostly a few weeks can be used to develop the stability indicating method which can be applied later for the analysis of samples generated from accelerated and long term stability studies. Further, kinetic study was also performed for different forced degradation parameters of the same combination, which help in determining order of reaction.

Keywords: amiloride hydrochloride, furosemide, kinetic study, stability indicating RP-HPLC method validation

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91 Cellular Technologies in Urology

Authors: R. Zhankina, U. Zhanbyrbekuly, A. Tamadon, M. Askarov, R. Sherkhanov, D. Akhmetov, D. Saipiyeva, N. Keulimzhaev

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Male infertility affects about 15% of couples of reproductive age. Approximately 10–15% have azoospermia who have previously been diagnosed with male infertility. Azoospermia is regarded as the absence of spermatozoa in the ejaculate and is found in 10-15% of infertile men. Non-obstructive azoospermia is considered a cause of male infertility that is not amenable to drug therapy. Patients with non-obstructive azoospermia are unable to have their "own" children and have only options for adoption or use of donor sperm. Advances in assisted reproductive technologies such as intracytoplasmic sperm injection in vitro fertilization have significantly changed the management of patients with non-obstructive azoospermia. Advances in biotechnology have increased the options for treating patients with non-obstructive azoospermia. Mesenchymal stem cell therapy has been recognized as a new option for infertility treatment. Material and methods of the study: After obtaining informed consent, 5 patients diagnosed with non-obstructive azoospermia were included in an open, non-randomized study. The age of the patients ranged from 24 to 35 years. The examination was carried out before the start of treatment, which included biochemical blood tests, hormonal profile levels (luteinizing hormone, follicle-stimulating hormone, testosterone, prolactin, inhibin B); tests for tumor markers; genetic research. All studies were carried out in compliance with the requirements of Protocol No. 8 dated 06/09/20, approved by the Local Ethical Commission of NJSC "Astana Medical University". The control examination of patients was carried out after 6 months, by re-taking the program and hormonal profile (testosterone, luteinizing hormone, follicle-stimulating hormone, prolactin, inhibin B). Before micro-TESE of the testis, all 5 patients underwent myeloexfusion in the operating room. During the micro-TESE, autotransplantation of mesenchymal stem cells into the testicular network, previously cultured in a cell technology laboratory for 2 weeks, was performed. Results of the study: in all patients, the levels of total testosterone increased, the level of follicle-stimulating hormone decreased, the levels of luteinizing hormone returned to normal, the level of inhibin B increased. IVF with a positive result; another patient (20%) had spermatogenesis cells. Non-obstructive azoospermia and mesenchymal stem cells Conclusions: The positive results of this work serve as the basis for the application of a new cellular therapeutic approach for the treatment of non-obstructive azoospermia using mesenchymal stem cells.

Keywords: cell therapy, regenerative medicine, male infertility, mesenchymal stem cells

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90 Aframomum melegueta Improves Antioxidant Status of Type 2 Diabetes Rats Model

Authors: Aminu Mohammed, Shahidul Islam

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Aframomum melegueta K.Schum commonly known as Grains of Paradise has been a popularly used spice in most of the African food preparation. Available data have shown that ethyl acetate fraction from crude ethanolic extract exhibited α-amylase and α-glucosidase inhibitory actions, improved pancreatic β-cell damage and ameliorated insulin resistance in diabetic rats. Additionally, 6-gingerol, 6-shogaol, 6-paradol and oleanolic acid are shown to be the compounds responsible for the antidiabetic action of A. melegueta. However, detail antioxidant potential of this spice in a diabetic animal model has not yet been reported. Thus, the present study investigates the effect of oral consumption of A. melegueta fruit on the in vivo antioxidant status of type 2 diabetes (T2D) model of rats. T2D was induced in rats by feeding a 10% fructose solution ad libitum for two weeks followed by a single intraperitoneal injection of streptozotocin (40 mg/kg body weight (bw)). The animals were orally administered with 150 (DAML) or 300 mg/kg bw (DAMH) of the fraction once daily for four weeks. Data were analyzed by using a statistical software package (SPSS for Windows, version 22, IBM Corporation, NY, USA) using Tukey’s-HSD multiple range post-hoc test. Values were considered significantly different at p < 0.05. According to the data, after four weeks of intervention, diabetic untreated animals showed significantly (p < 0.05) elevation of blood glucose levels. The levels of thiobarbituric acid reactive substances (TBARS) were observed to increase with concomitant reduction of reduced glutathione (GSH) levels in the serum and organs (liver, kidney, heart and pancreas) of diabetic untreated animals. The activities of endogenous antioxidant enzymes (superoxide dismutase, catalase, glutathione peroxidase, and reductase) were greatly reduced in the serum and organs of diabetic untreated animals compared to the normal animals. These alterations were reverted to near-normal after the treatment of A. melegueta fruit in the treated groups (DAML & DAMH) within the study period, especially at the dose of 300 mg/kg bw. This potent antioxidant action may partly be attributed to the presence of the 6-Gingerol, 6-shogaol and 6-paradol are known to possess antioxidant action. The results of our study showed that A. melegueta intake improved the antioxidant status of T2D rats and therefore could be used to ameliorate the diabetes-induced oxidative damage.

Keywords: Aframomum melegueta, antioxidant, ethyl acetate extract, type 2 diabetes

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89 Experimental Investigation of Hydrogen Addition in the Intake Air of Compressed Engines Running on Biodiesel Blend

Authors: Hendrick Maxil Zárate Rocha, Ricardo da Silva Pereira, Manoel Fernandes Martins Nogueira, Carlos R. Pereira Belchior, Maria Emilia de Lima Tostes

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This study investigates experimentally the effects of hydrogen addition in the intake manifold of a diesel generator operating with a 7% biodiesel-diesel oil blend (B7). An experimental apparatus setup was used to conduct performance and emissions tests in a single cylinder, air cooled diesel engine. This setup consisted of a generator set connected to a wirewound resistor load bank that was used to vary engine load. In addition, a flowmeter was used to determine hydrogen volumetric flowrate and a digital anemometer coupled with an air box to measure air flowrate. Furthermore, a digital precision electronic scale was used to measure engine fuel consumption and a gas analyzer was used to determine exhaust gas composition and exhaust gas temperature. A thermopar was installed near the exhaust collection to measure cylinder temperature. In-cylinder pressure was measured using an AVL Indumicro data acquisition system with a piezoelectric pressure sensor. An AVL optical encoder was installed in the crankshaft and synchronized with in-cylinder pressure in real time. The experimental procedure consisted of injecting hydrogen into the engine intake manifold at different mass concentrations of 2,6,8 and 10% of total fuel mass (B7 + hydrogen), which represented energy fractions of 5,15, 20 and 24% of total fuel energy respectively. Due to hydrogen addition, the total amount of fuel energy introduced increased and the generators fuel injection governor prevented any increases of engine speed. Several conclusions can be stated from the test results. A reduction in specific fuel consumption as a function of hydrogen concentration increase was noted. Likewise, carbon dioxide emissions (CO2), carbon monoxide (CO) and unburned hydrocarbons (HC) decreased as hydrogen concentration increased. On the other hand, nitrogen oxides emissions (NOx) increased due to average temperatures inside the cylinder being higher. There was also an increase in peak cylinder pressure and heat release rate inside the cylinder, since the fuel ignition delay was smaller due to hydrogen content increase. All this indicates that hydrogen promotes faster combustion and higher heat release rates and can be an important additive to all kind of fuels used in diesel generators.

Keywords: diesel engine, hydrogen, dual fuel, combustion analysis, performance, emissions

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88 Microbiological Analysis on Anatomical Specimens of Cats for Use in Veterinary Surgery

Authors: Raphael C. Zero, Marita V. Cardozo, Thiago A. S. S. Rocha, Mariana T. Kihara, Fernando A. Ávila, Fabrício S. Oliveira

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There are several fixative and preservative solutions for use on cadavers, many of them using formaldehyde as the fixative or anatomical part preservative. In some countries, such as Brazil, this toxic agent has been increasingly restricted. The objective of this study was to microbiologically identify and quantify the key agents in tanks containing 96GL ethanol or sodium chloride solutions, used respectively as fixatives and preservatives of cat cadavers. Eight adult cat corpses, three females and five males, with an average weight of 4.3 kg, were used. After injection via the external common carotid artery (120 ml/kg, 95% 96GL ethyl alcohol and 5% pure glycerin), the cadavers were fixed in a plastic tank with 96GL ethanol for 60 days. After fixing, they were stored in a 30% sodium chloride aqueous solution for 120 days in a similar tank. Samples were collected at the start of the experiment - before the animals were placed in the ethanol tanks, and monthly thereafter. The bacterial count was performed by Pour Plate Method in BHI agar (Brain Heart Infusion) and the plates were incubated aerobically and anaerobically for 24h at 37ºC. MacConkey agar, SPS agar (Sulfite Polymyxin Sulfadizine) and MYP Agar Base were used to isolate the microorganisms. There was no microbial growth in the samples prior to alcohol fixation. After 30 days of fixation in the alcohol solution, total aerobic and anaerobic (<1.0 x 10 CFU/ml) were found and Pseudomonas sp., Staphylococcus sp., Clostridium sp. were the identified agents. After 60 days in the alcohol fixation solution, total aerobes (<1.0 x 10 CFU/ml) and total anaerobes (<2.2 x 10 CFU/mL) were found, and the identified agents were the same. After 30 days of storage in the aqueous solution of 30% sodium chloride, total aerobic (<5.2 x 10 CFU/ml) and total anaerobes (<3.7 x 10 CFU/mL) were found and the agents identified were Staphylococcus sp., Clostridium sp., and fungi. After 60 days of sodium chloride storage, total aerobic (<3.0 x 10 CFU / ml) and total anaerobes (<7.0 x 10 CFU/mL) were found and the identified agents remained the same: Staphylococcus sp., Clostridium sp., and fungi. The microbiological count was low and visual inspection did not reveal signs of contamination in the tanks. There was no strong odor or purification, which proved the technique to be microbiologically effective in fixing and preserving the cat cadavers for the four-month period in which they are provided to undergraduate students of University of Veterinary Medicine for surgery practice. All experimental procedures were approved by the Municipal Legal Department (protocol 02.2014.000027-1). The project was funded by FAPESP (protocol 2015-08259-9).

Keywords: anatomy, fixation, microbiology, small animal, surgery

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87 Efficacy and Safety of COVID-19 Vaccination in Patients with Multiple Sclerosis: Looking Forward to Post-COVID-19

Authors: Achiron Anat, Mathilda Mandel, Mayust Sue, Achiron Reuven, Gurevich Michael

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Introduction: As coronavirus disease 2019 (COVID-19) vaccination is currently spreading around the world, it is of importance to assess the ability of multiple sclerosis (MS) patients to mount an appropriate immune response to the vaccine in the context of disease-modifying treatments (DMT’s). Objectives: Evaluate immunity generated following COVID-19 vaccination in MS patients, and assess factors contributing to protective humoral and cellular immune responses in MS patients vaccinated against severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) virus infection. Methods: Review our recent data related to (1) the safety of PfizerBNT162b2 COVID-19 mRNA vaccine in adult MS patients; (2) the humoral post-vaccination SARS-CoV2 IgG response in MS vaccinees using anti-spike protein-based serology; and (3) the cellular immune response of memory B-cells specific for SARS-CoV-2 receptor-binding domain (RBD) and memory T-cells secreting IFN-g and/or IL-2 in response to SARS-CoV2 peptides using ELISpot/Fluorospot assays in MS patients either untreated or under treatment with fingolimod, cladribine, or ocrelizumab; (4) covariate parameters related to mounting protective immune responses. Results: COVID-19 vaccine proved safe in MS patients, and the adverse event profile was mainly characterised by pain at the injection site, fatigue, and headache. Not any increased risk of relapse activity was noted and the rate of patients with acute relapse was comparable to the relapse rate in non-vaccinated patients during the corresponding follow-up period. A mild increase in the rate of adverse events was noted in younger MS patients, among patients with lower disability, and in patients treated with DMTs. Following COVID-19 vaccination protective humoral immune response was significantly decreased in fingolimod- and ocrelizumab- treated MS patients. SARS-CoV2 specific B-cell and T-cell cellular responses were respectively decreased. Untreated MS patients and patients treated with cladribine demonstrated protective humoral and cellular immune responses, similar to healthy vaccinated subjects. Conclusions: COVID-19 BNT162b2 vaccine proved as safe for MS patients. No increased risk of relapse activity was noted post-vaccination. Although COVID-19 vaccination is new, accumulated data demonstrate differences in immune responses under various DMT’s. This knowledge can help to construct appropriate COVID-19 vaccine guidelines to ensure proper immune responses for MS patients.

Keywords: covid-19, vaccination, multiple sclerosis, IgG

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86 Numerical Investigation of the Effects of Surfactant Concentrations on the Dynamics of Liquid-Liquid Interfaces

Authors: Bamikole J. Adeyemi, Prashant Jadhawar, Lateef Akanji

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Theoretically, there exist two mathematical interfaces (fluid-solid and fluid-fluid) when a liquid film is present on solid surfaces. These interfaces overlap if the mineral surface is oil-wet or mixed wet, and therefore, the effects of disjoining pressure are significant on both boundaries. Hence, dewetting is a necessary process that could detach oil from the mineral surface. However, if the thickness of the thin water film directly in contact with the surface is large enough, disjoining pressure can be thought to be zero at the liquid-liquid interface. Recent studies show that the integration of fluid-fluid interactions with fluid-rock interactions is an important step towards a holistic approach to understanding smart water effects. Experiments have shown that the brine solution can alter the micro forces at oil-water interfaces, and these ion-specific interactions lead to oil emulsion formation. The natural emulsifiers present in crude oil behave as polyelectrolytes when the oil interfaces with low salinity water. Wettability alteration caused by low salinity waterflooding during Enhanced Oil Recovery (EOR) process results from the activities of divalent ions. However, polyelectrolytes are said to lose their viscoelastic property with increasing cation concentrations. In this work, the influence of cation concentrations on the dynamics of viscoelastic liquid-liquid interfaces is numerically investigated. The resultant ion concentrations at the crude oil/brine interfaces were estimated using a surface complexation model. Subsequently, the ion concentration parameter is integrated into a mathematical model to describe its effects on the dynamics of a viscoelastic interfacial thin film. The film growth, stability, and rupture were measured after different time steps for three types of fluids (Newtonian, purely elastic and viscoelastic fluids). The interfacial films respond to exposure time in a similar manner with an increasing growth rate, which resulted in the formation of more droplets with time. Increased surfactant accumulation at the interface results in a higher film growth rate which leads to instability and subsequent formation of more satellite droplets. Purely elastic and viscoelastic properties limit film growth rate and consequent film stability compared to the Newtonian fluid. Therefore, low salinity and reduced concentration of the potential determining ions in injection water will lead to improved interfacial viscoelasticity.

Keywords: liquid-liquid interfaces, surfactant concentrations, potential determining ions, residual oil mobilization

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85 Transgenerational Impact of Intrauterine Hyperglycaemia to F2 Offspring without Pre-Diabetic Exposure on F1 Male Offspring

Authors: Jun Ren, Zhen-Hua Ming, He-Feng Huang, Jian-Zhong Sheng

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Adverse intrauterine stimulus during critical or sensitive periods in early life, may lead to health risk not only in later life span, but also further generations. Intrauterine hyperglycaemia, as a major feature of gestational diabetes mellitus (GDM), is a typical adverse environment for both F1 fetus and F1 gamete cells development. However, there is scare information of phenotypic difference of metabolic memory between somatic cells and germ cells exposed by intrauterine hyperglycaemia. The direct transmission effect of intrauterine hyperglycaemia per se has not been assessed either. In this study, we built a GDM mice model and selected male GDM offspring without pre-diabetic phenotype as our founders, to exclude postnatal diabetic influence on gametes, thereby investigate the direct transmission effect of intrauterine hyperglycaemia exposure on F2 offspring, and we further compared the metabolic difference of affected F1-GDM male offspring and F2 offspring. A GDM mouse model of intrauterine hyperglycemia was established by intraperitoneal injection of streptozotocin after pregnancy. Pups of GDM mother were fostered by normal control mothers. All the mice were fed with standard food. Male GDM offspring without metabolic dysfunction phenotype were crossed with normal female mice to obtain F2 offspring. Body weight, glucose tolerance test, insulin tolerance test and homeostasis model of insulin resistance (HOMA-IR) index were measured in both generations at 8 week of age. Some of F1-GDM male mice showed impaired glucose tolerance (p < 0.001), none of F1-GDM male mice showed impaired insulin sensitivity. Body weight of F1-GDM mice showed no significance with control mice. Some of F2-GDM offspring exhibited impaired glucose tolerance (p < 0.001), all the F2-GDM offspring exhibited higher HOMA-IR index (p < 0.01 of normal glucose tolerance individuals vs. control, p < 0.05 of glucose intolerance individuals vs. control). All the F2-GDM offspring exhibited higher ITT curve than control (p < 0.001 of normal glucose tolerance individuals, p < 0.05 of glucose intolerance individuals, vs. control). F2-GDM offspring had higher body weight than control mice (p < 0.001 of normal glucose tolerance individuals, p < 0.001 of glucose intolerance individuals, vs. control). While glucose intolerance is the only phenotype that F1-GDM male mice may exhibit, F2 male generation of healthy F1-GDM father showed insulin resistance, increased body weight and/or impaired glucose tolerance. These findings imply that intrauterine hyperglycaemia exposure affects germ cells and somatic cells differently, thus F1 and F2 offspring demonstrated distinct metabolic dysfunction phenotypes. And intrauterine hyperglycaemia exposure per se has a strong influence on F2 generation, independent of postnatal metabolic dysfunction exposure.

Keywords: inheritance, insulin resistance, intrauterine hyperglycaemia, offspring

Procedia PDF Downloads 238
84 Effect of Starch and Plasticizer Types and Fiber Content on Properties of Polylactic Acid/Thermoplastic Starch Blend

Authors: Rangrong Yoksan, Amporn Sane, Nattaporn Khanoonkon, Chanakorn Yokesahachart, Narumol Noivoil, Khanh Minh Dang

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Polylactic acid (PLA) is the most commercially available bio-based and biodegradable plastic at present. PLA has been used in plastic related industries including single-used containers, disposable and environmentally friendly packaging owing to its renewability, compostability, biodegradability, and safety. Although PLA demonstrates reasonably good optical, physical, mechanical, and barrier properties comparable to the existing petroleum-based plastics, its brittleness and mold shrinkage as well as its price are the points to be concerned for the production of rigid and semi-rigid packaging. Blending PLA with other bio-based polymers including thermoplastic starch (TPS) is an alternative not only to achieve a complete bio-based plastic, but also to reduce the brittleness, shrinkage during molding and production cost of the PLA-based products. TPS is a material produced mainly from starch which is cheap, renewable, biodegradable, compostable, and non-toxic. It is commonly prepared by a plasticization of starch under applying heat and shear force. Although glycerol has been reported as one of the most plasticizers used for preparing TPS, its migration caused the surface stickiness of the TPS products. In some cases, mixed plasticizers or natural fibers have been applied to impede the retrogradation of starch or reduce the migration of glycerol. The introduction of fibers into TPS-based materials could reinforce the polymer matrix as well. Therefore, the objective of the present research is to study the effect of starch type (i.e. native starch and phosphate starch), plasticizer type (i.e. glycerol and xylitol with a weight ratio of glycerol to xylitol of 100:0, 75:25, 50:50, 25:75, and 0:100), and fiber content (i.e. in the range of 1-25 % wt) on properties of PLA/TPS blend and composite. PLA/TPS blends and composites were prepared using a twin-screw extruder and then converted into dumbbell-shaped specimens using an injection molding machine. The PLA/TPS blends prepared by using phosphate starch showed higher tensile strength and stiffness than the blends prepared by using the native one. In contrast, the blends from native starch exhibited higher extensibility and heat distortion temperature (HDT) than those from the modified starch. Increasing xylitol content resulted in enhanced tensile strength, stiffness, and water resistance, but decreased extensibility and HDT of the PLA/TPS blend. Tensile properties and hydrophobicity of the blend could be improved by incorporating silane treated-jute fibers.

Keywords: polylactic acid, thermoplastic starch, Jute fiber, composite, blend

Procedia PDF Downloads 424
83 Through the Robot’s Eyes: A Comparison of Robot-Piloted, Virtual Reality, and Computer Based Exposure for Fear of Injections

Authors: Bonnie Clough, Tamara Ownsworth, Vladimir Estivill-Castro, Matt Stainer, Rene Hexel, Andrew Bulmer, Wendy Moyle, Allison Waters, David Neumann, Jayke Bennett

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The success of global vaccination programs is reliant on the uptake of vaccines to achieve herd immunity. Yet, many individuals do not obtain vaccines or venipuncture procedures when needed. Whilst health education may be effective for those individuals who are hesitant due to safety or efficacy concerns, for many of these individuals, the primary concern relates to blood or injection fear or phobia (BII). BII is highly prevalent and associated with a range of negative health impacts, both at individual and population levels. Exposure therapy is an efficacious treatment for specific phobias, including BII, but has high patient dropout and low implementation by therapists. Whilst virtual reality approaches exposure therapy may be more acceptable, they have similarly low rates of implementation by therapists and are often difficult to tailor to an individual client’s needs. It was proposed that a piloted robot may be able to adequately facilitate fear induction and be an acceptable approach to exposure therapy. The current study examined fear induction responses, acceptability, and feasibility of a piloted robot for BII exposure. A Nao humanoid robot was programmed to connect with a virtual reality head-mounted display, enabling live streaming and exploration of real environments from a distance. Thirty adult participants with BII fear were randomly assigned to robot-pilot or virtual reality exposure conditions in a laboratory-based fear exposure task. All participants also completed a computer-based two-dimensional exposure task, with an order of conditions counterbalanced across participants. Measures included fear (heart rate variability, galvanic skin response, stress indices, and subjective units of distress), engagement with a feared stimulus (eye gaze: time to first fixation and a total number of fixations), acceptability, and perceived treatment credibility. Preliminary results indicate that fear responses can be adequately induced via a robot-piloted platform. Further results will be discussed, as will implications for the treatment of BII phobia and other fears. It is anticipated that piloted robots may provide a useful platform for facilitating exposure therapy, being more acceptable than in-vivo exposure and more flexible than virtual reality exposure.

Keywords: anxiety, digital mental health, exposure therapy, phobia, robot, virtual reality

Procedia PDF Downloads 78
82 Beneficial Effect of Autologous Endometrial Stromal Cell Co-Culture on Day 3 Embryo Quality

Authors: I. Bochev, A. Shterev, S. Kyurkchiev

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One of the factors associated with poor success rates in human in vitro fertilization (IVF) is the suboptimal culture conditions in which fertilization and early embryonic growth occur. Co-culture systems with helper cell lines appear to enhance the in vitro conditions and allow embryos to demonstrate improved in vitro development. The co-culture of human embryos with monolayers of autologous endometrial stromal cell (EnSCs) results in increased blastocyst development with a larger number of blastomeres, lower incidence of fragmentation and higher pregnancy rates in patients with recurrent implantation failure (RIF). The aim of the study was to examine the influence of autologous endometrial stromal cell (EnSC) co-culture on day 3 embryo quality by comparing the morphological status of the embryos from the same patients undergoing consecutive IVF/Intracytoplasmic sperm injection (ICSI) cycles without and with EnSC co-culture. This retrospective randomized study (2015-2017) includes 20 couples and a total of 46 IVF/ICSI cycles. Each patient couple included had at least two IVF/ICSI procedures – one with and one without autologous EnSC co-culture. Embryo quality was assessed at 68±1 hours in culture, according to Istanbul consensus criteria (2010). Day 3 embryos were classified into three groups: good – grade 1; fair – grade 2; poor – grade 3. Embryos from all cycles were divided into two groups (A – co-cultivated; B – not co-cultivated) and analyzed. Second, for each patient couple, embryos from matched IVF/ICSI cycles (with and without co-culture) were analyzed separately. When an analysis of co-cultivated day 3 embryos from all cycles was performed (n=137; group A), 43.1% of the embryos were graded as “good”, which was not significantly different from the respective embryo quality rate of 42.2% (p = NS) in group B (n=147) with non-co-cultivated embryos. The proportions of fair and poor quality embryos in group A and group B were similar as well – 11.7% vs 10.2% and 45.2% vs 47.6% (p=NS), respectively. Nevertheless, the separate embryo analysis by matched cycles for each couple revealed that in 65% of the cases the proportion of morphologically better embryos was increased in cycles with co-culture in comparison with those without co-culture. A decrease in this proportion after endometrial stromal cell co-cultivation was found in 30% of the cases, whereas no difference was observed in only one couple. The results demonstrated that there is no marked difference in the overall morphological quality between co-cultured and non-co-cultured embryos on day 3. However, in significantly greater percentage of couples the process of autologous EnSC co-culture could increase the proportion of morphologically improved day 3 embryos. By mimicking the in vivo relationship between embryo and maternal environment, co-culture in autologous EnSC system represents a perspective approach to improve the quality of embryos in cases with elevated risk for development of embryos with impaired morphology.

Keywords: autologous endometrial stromal cells, co-culture, day 3 embryo, morphological quality

Procedia PDF Downloads 234
81 Effect of Accelerated Aging on Antibacterial and Mechanical Properties of SEBS Compounds

Authors: Douglas N. Simoes, Michele Pittol, Vanda F. Ribeiro, Daiane Tomacheski, Ruth M. C. Santana

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Thermoplastic elastomers (TPE) compounds are used in a wide range of applications, like home appliances, automotive components, medical devices, footwear, and others. These materials are susceptible to microbial attack, causing a crack in polymer chains compounds based on SEBS copolymers, poly (styrene-b-(ethylene-co-butylene)-b-styrene, are a class of TPE, largely used in domestic appliances like refrigerator seals (gaskets), bath mats and sink squeegee. Moisture present in some areas (such as shower area and sink) in addition to organic matter provides favorable conditions for microbial survival and proliferation, contributing to the spread of diseases besides the reduction of product life cycle due the biodegradation process. Zinc oxide (ZnO) has been studied as an alternative antibacterial additive due its biocidal effect. It is important to know the influence of these additives in the properties of the compounds, both at the beginning and during the life cycle. In that sense, the aim of this study was to evaluate the effect of accelerated aging in oven on antibacterial and mechanical properties of ZnO loaded SEBS based TPE compounds. Two different comercial zinc oxide, named as WR and Pe were used in proportion of 1%. A compound with no antimicrobial additive (standard) was also tested. The compounds were prepared using a co-rotating double screw extruder (L/D ratio of 40/1 and 16 mm screw diameter). The extrusion parameters were kept constant for all materials, screw rotation rate was set at 226 rpm, with a temperature profile from 150 to 190 ºC. Test specimens were prepared using the injection molding machine at 190 ºC. The Standard Test Method for Rubber Property—Effect of Liquids was applied in order to simulate the exposition of TPE samples to detergent ingredients during service. For this purpose, ZnO loaded TPE samples were immersed in a 3.0% w/v detergent (neutral) and accelerated aging in oven at 70°C for 7 days. Compounds were characterized by changes in mechanical (hardness and tension properties) and mass. The Japan Industrial Standard (JIS) Z 2801:2010 was applied to evaluate antibacterial properties against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli). The microbiological tests showed a reduction up to 42% in E. coli and up to 49% in S. aureus population in non-aged samples. There were observed variations in elongation and hardness values with the addition of zinc The changes in tensile at rupture and mass were not significant between non-aged and aged samples.

Keywords: antimicrobial, domestic appliance, sebs, zinc oxide

Procedia PDF Downloads 247
80 Insertion of Photovoltaic Energy at Residential Level at Tegucigalpa and Comayagüela, Honduras

Authors: Tannia Vindel, Angel Matute, Erik Elvir, Kelvin Santos

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Currently in Honduras, is been incentivized the generation of energy using renewable fonts, such as: hydroelectricity, wind power, biomass and, more recently with the strongest growth, photovoltaic energy. In July 2015 were installed 455.2 MW of photovoltaic energy, increasing by 24% the installed capacity of the national interconnected system existing in 2014, according the National Energy Company (NEC), that made possible reduce the thermoelectric dependency of the system. Given the good results of those large-scale photovoltaic plants, arises the question: is it interesting for the distribution utility and for the consumers the integration of photovoltaic systems in micro-scale in the urban and rural areas? To answer that question has been researched the insertion of photovoltaic energy in the residential sector in Tegucigalpa and Comayagüela (Central District), Honduras to determine the technical and economic viability. Francisco Morazán department, according the National Statistics Institute (NSI), in 2001 had more than 180,000 houses with power service. Tegucigalpa, department and Honduras capital, and Comayagüela, both, have the highest population density in the region, with 1,300,000 habitants in 2014 (NSI). The residential sector in the south-central region of Honduras represents a high percentage being 49% of total consumption, according with NEC in 2014; where 90% of this sector consumes in a range of 0 to 300 kWh / month. All this, in addition to the high level of losses in the transmission and distribution systems, 31.3% in 2014, and the availability of an annual average solar radiation of 5.20 kWh/(m2∙day) according to the NASA, suggests the feasibility of the implementation of photovoltaic systems as a solution to give a level of independency to the households, and besides could be capable of injecting the non-used energy to the grid. The capability of exchange of energy with the grid could make the photovoltaic systems acquisition more affordable to the consumers, because of the compensation energy programs or other kinds of incentives that could be created. Technical viability of the photovoltaic systems insertion has been analyzed, considering the solar radiation monthly average to determine the monthly average of energy that would be generated with the technology accessible locally and the effects of the injection of the energy locally generated on the grid. In addition, the economic viability has been analyzed too, considering the photovoltaic systems high costs, costs of the utility, location and monthly energy consumption requirements of the families. It was found that the inclusion of photovoltaic systems in Tegucigalpa and Comayagüela could decrease in 6 MW the demand for the region if 100% of the households use photovoltaic systems, which acquisition may be more accessible with the help of government incentives and/or the application of energy exchange programs.

Keywords: grid connected, photovoltaic, residential, technical analysis

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79 Effect of Fuel Type on Design Parameters and Atomization Process for Pressure Swirl Atomizer and Dual Orifice Atomizer for High Bypass Turbofan Engine

Authors: Mohamed K. Khalil, Mohamed S. Ragab

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Atomizers are used in many engineering applications including diesel engines, petrol engines and spray combustion in furnaces as well as gas turbine engines. These atomizers are used to increase the specific surface area of the fuel, which achieve a high rate of fuel mixing and evaporation. In all combustion systems reduction in mean drop size is a challenge which has many advantages since it leads to rapid and easier ignition, higher volumetric heat release rate, wider burning range and lower exhaust concentrations of the pollutant emissions. Pressure atomizers have a different configuration for design such as swirl atomizer (simplex), dual orifice, spill return, plain orifice, duplex and fan spray. Simplex pressure atomizers are the most common type of all. Among all types of atomizers, pressure swirl types resemble a special category since they differ in quality of atomization, the reliability of operation, simplicity of construction and low expenditure of energy. But, the disadvantages of these atomizers are that they require very high injection pressure and have low discharge coefficient owing to the fact that the air core covers the majority of the atomizer orifice. To overcome these problems, dual orifice atomizer was designed. This paper proposes a detailed mathematical model design procedure for both pressure swirl atomizer (Simplex) and dual orifice atomizer, examines the effects of varying fuel type and makes a clear comparison between the two types. Using five types of fuel (JP-5, JA1, JP-4, Diesel and Bio-Diesel) as a case study, reveal the effect of changing fuel type and its properties on atomizers design and spray characteristics. Which effect on combustion process parameters; Sauter Mean Diameter (SMD), spray cone angle and sheet thickness with varying the discharge coefficient from 0.27 to 0.35 during takeoff for high bypass turbofan engines. The spray atomizer performance of the pressure swirl fuel injector was compared to the dual orifice fuel injector at the same differential pressure and discharge coefficient using Excel. The results are analyzed and handled to form the final reliability results for fuel injectors in high bypass turbofan engines. The results show that the Sauter Mean Diameter (SMD) in dual orifice atomizer is larger than Sauter Mean Diameter (SMD) in pressure swirl atomizer, the film thickness (h) in dual orifice atomizer is less than the film thickness (h) in pressure swirl atomizer. The Spray Cone Angle (α) in pressure swirl atomizer is larger than Spray Cone Angle (α) in dual orifice atomizer.

Keywords: gas turbine engines, atomization process, Sauter mean diameter, JP-5

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78 Leuprolide Induced Scleroderma Renal Crisis: A Case Report

Authors: Nirali Sanghavi, Julia Ash, Amy Wasserman

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Introduction: To the best of our knowledge, there is only one case report that found an association between leuprolide and scleroderma renal crisis (SRC). Leuprolide has been noted to cause acute renal failure in some patients. Given the close timing of the leuprolide injection and the worsening renal function in our patient, leuprolide likely caused exacerbation of lupus nephritis and SRC. Interestingly, our patient on long-term hydroxychloroquine (HCQ) with normal baseline cardiac function was found to have HCQ cardiomyopathy highlighting the need for close monitoring of HCQ toxicity. We know that some of the risk factors that are involved in HCQ induced cardiomyopathy are older age, females, increased dose and >10 years of HCQ use, and pre-existing cardiac and renal insufficiency. Case presentation: A 34-year-old African American woman with a history of overlap of systemic lupus erythematosus (SLE) and scleroderma features and class III lupus nephritis presented with severe headaches, elevated blood pressure (180/120 mmHg) and worsening creatinine levels (2.07 mg/dL). The headaches started 1 month ago after she started leuprolide injections for fibroids. She was being treated with mycophenolate mofetil 1 gm twice a day, belimumab weekly, HCQ 200mg, and prednisone 5 mg daily. She has been on HCQ since her teenage years. The examination was unremarkable except for proximal interphalangeal joint contractures in the right hand and sclerodactyly of bilateral hands, unchanged from baseline. Laboratory findings include urinalysis, which showed 3+ protein, 1+ blood, 6 red blood cells, and 14 white blood cells ruling out thrombotic microangiopathy. C3 was 32 mg/dL, C4 <5 mg/dL, and +dsDNA increased >1000. She was started on captopril and discharged once creatinine and blood pressure was controlled. She was readmitted with hypertension, hyperkalemia, worsening creatinine, nephrotic range proteinuria, complaints of chest pressure, and shortness of breath with pleuritic chest pain. Physical examination and lab findings were unchanged. She was treated with pulse dose methyl prednisone followed by taper and multiple anti-hypertensive agents, including captopril, for presumed lupus nephritis flare versus SRC. Renal biopsy was consistent with SRC and class IV lupus nephritis and was started on cyclophosphamide. While cardiac biopsy showed borderline myocarditis without necrosis and cytoplasmic vacuolization consistent with HCQ cardiomyopathy, hence HCQ was discontinued. Summary: It highlights a rare association of leuprolide causing exacerbation of lupus nephritis or SRC. Although rare, the current case reinforces the importance of close monitoring for HCQ toxicity in patients with renal insufficiency.

Keywords: leuprolide, lupus nephritis, scleroderma, SLE

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77 Comparative Histological, Immunohistochemical and Biochemical Study on the Effect of Vit. C, Vit. E, Gallic Acid and Silymarin on Carbon Tetrachloride Model of Liver Fibrosis in Rats

Authors: Safaa S. Hassan, Mohammed H. Elbakry, Safwat A. Mangoura, Zainab M. Omar

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Background: Liver fibrosis is the main reason for increased mortality in chronic liver disease. It has no standard treatment. Antioxidants from a variety of sources are capable of slowing or preventing oxidation of other molecules. Aim: to evaluate the hepatoprotective effect of vit. C, vit. E and gallic acid in comparison to silymarin in the rat model of carbon tetrachloride induced liver fibrosis and their possible mechanisms of action. Material& Methods: A total number of 60 adult male albino rats 160-200gm were divided into six equal groups; received subcutaneous (s.c) injection for 8 weeks. Group I: as control. Group II: received 1.5 mL/kg of CCL4 .Group III: CCL4 and co- treatment with silymarin 100mg/kg p.o. daily. Group IV: CCL4 and co-treatment with vit. C 50mg/kg p.o. daily. Group V: CCL4 and co-treatment with vit. E 200mg/kg. p.o. Group VI: CCL4 and co-treatment with Gallic acid 100mg/kg. p.o. daily. Liver was processed for histological and immunohistochemical examination. Levels of AST, ALT, ALP, reduced GSH, MDA, SOD and hydroxyproline concentration were measured and evaluated statistically. Results: Light and electron microscopic examination of liver of group II exhibited foci of altered cells with dense nuclei and vacuolated, granular cytoplasm, mononuclear cell infiltration in portal areas, profuse collagen fiber deposits were found around portal tract, more intense staining α-SMA-positive cells occupied most of the liver fibrosis tissue, electron lucent areas in the cytoplasm of the hepatocytes, margination of nuclear chromatin. Treatment by any of the antioxidants variably reduced the hepatic structural changes induced by CCL4. Biochemical analysis showed that carbon tetrachloride significantly increased the levels of serum AST, ALT, ALP, hepatic malondialdehyde and hydroxyproline content. Moreover, it decreased the activities of superoxide dismutase and glutathione. Treatment with silymarin, gallic acid, vit. C and vit. E decreased significantly the AST, ALT, and ALP levels in plasma, MDA and hydroxyproline and increased the activities of SOD and glutathione in liver tissue. The effect of administration of CCl4 was improved with the used antioxidants in variable degrees. The most efficient antioxidant was silymarin followed by gallic acid and vit. C then vit. E. It is possibly due to their antioxidant effect, free radical scavenging properties and the reduction of oxidant dependent activation and proliferation of HSCs. Conclusion: So these antioxidants can be a promising drugs candidate for ameliorating liver fibrosis better than the use of the drugs and their side effects.

Keywords: antioxidant, ccl4, gallic acid, liver fibrosis

Procedia PDF Downloads 273
76 Anti-Apoptotic Effect of Pueraria tuberosa in Rats with Streptozotocin Induced Diabetic Nephropathy

Authors: Rashmi Shukla, Yamini Bhusan Tripathi

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Diabetic nephropathy (DN) is characterized as diabetic kidney disease which involves many pathways e.g. hyperactivated protein kinase c (PKC), polyol pathway, excess production of advanced glycation end product (AGEs) & free radical accumulation etc. All of them results to hypoxia followed by apoptosis of podocytes, glomerulosclerosis, extracellular matrix (ECM) accumulation and fibrosis resulting to irreversible changes in kidney. This is continuously rising worldwide and there are not enough specific drugs, to retard its progress. Due to increasing side effects of allopathic drugs, interest in herbal remedies is growing. Earlier, we have reported that PTY-2 (a phytomedicine, derived from Pueraria tuberosa Linn.) inhibits the accumulation of extracellular matrix (ECM) through activation of MMP-9. Present study exhibited the therapeutic potential of Pueraria tuberosa in the prevention of podocytes apoptosis and modulation of nephrin expression in streptozotocin (STZ) induced DN rats. DN rats were produced by maintaining persistent hyperglycemia for 8 weeks by intra-peritoneal injection of 55 mg/kg streptozotocin (STZ). These rats were randomly divided in 2 groups, i.e. DN control, and DN+ water extract of Pueraria tuberosa (PTW). One group of age-matched normal rats served as non-diabetic control (group-1), The STZ induced DN rats (group-2) and DN+PTW treated rats (group-3). The PTW was orally administered (0.3g/kg) daily to group-2 rats and drug vector (1 ml of 10% tween 20) in control rats. The treatments were continued for 20 days and blood and urine samples were collected. Rats were then sacrificed to investigate the expression Bcl2, Bax and nephroprotective protein i.e. nephrin in kidney glomerulus. The effect of PTW was evaluated, we have found that the PTW significantly(p < .001) reversed the raised serum urea, serum creatinine, urine protein and improved the creatinine clearance in STZ induce diabetic nephropathy in rats and also significantly(p < .001) prevented the rise in urine albumin excretion. The Western blot analysis of kidney tissue homogenate showed increased expression of Bcl2 in PTW treated rats. The RT-PCR showed the increased expression and accumulation of nephrin mRNA. The confocal photomicrographs also supported the reduction of Bax and a simultaneous increase in Bcl2 and nephrin in glomerular podocytes. Hence, our finding suggests that the nephroprotective role of PTW is mediated via restoration of nephrin thus prevents the podocytes apoptosis and ameliorates diabetic nephropathy. The clinical trial of PTW would prove to be a potential food supplement/ drug of alternative medicine for patients with diabetic nephropathy in early stage.

Keywords: Pueraria tuberosa, diabetic nephropathy, anti-apoptosis, nephrin

Procedia PDF Downloads 219
75 Analytical Tools for Multi-Residue Analysis of Some Oxygenated Metabolites of PAHs (Hydroxylated, Quinones) in Sediments

Authors: I. Berger, N. Machour, F. Portet-Koltalo

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Polycyclic aromatic hydrocarbons (PAHs) are toxic and carcinogenic pollutants produced in majority by incomplete combustion processes in industrialized and urbanized areas. After being emitted in atmosphere, these persistent contaminants are deposited to soils or sediments. Even if persistent, some can be partially degraded (photodegradation, biodegradation, chemical oxidation) and they lead to oxygenated metabolites (oxy-PAHs) which can be more toxic than their parent PAH. Oxy-PAHs are less measured than PAHs in sediments and this study aims to compare different analytical tools in order to extract and quantify a mixture of four hydroxylated PAHs (OH-PAHs) and four carbonyl PAHs (quinones) in sediments. Methodologies: Two analytical systems – HPLC with on-line UV and fluorescence detectors (HPLC-UV-FLD) and GC coupled to a mass spectrometer (GC-MS) – were compared to separate and quantify oxy-PAHs. Microwave assisted extraction (MAE) was optimized to extract oxy-PAHs from sediments. Results: First OH-PAHs and quinones were analyzed in HPLC with on-line UV and fluorimetric detectors. OH-PAHs were detected with the sensitive FLD, but the non-fluorescent quinones were detected with UV. The limits of detection (LOD)s obtained were in the range (2-3)×10-4 mg/L for OH-PAHs and (2-3)×10-3 mg/L for quinones. Second, even if GC-MS is not well adapted to the analysis of the thermodegradable OH-PAHs and quinones without any derivatization step, it was used because of the advantages of the detector in terms of identification and of GC in terms of efficiency. Without derivatization, only two of the four quinones were detected in the range 1-10 mg/L (LODs=0.3-1.2 mg/L) and LODs were neither very satisfying for the four OH-PAHs (0.18-0.6 mg/L). So two derivatization processes were optimized, comparing to literature: one for silylation of OH-PAHs, one for acetylation of quinones. Silylation using BSTFA/TCMS 99/1 was enhanced using a mixture of catalyst solvents (pyridine/ethyle acetate) and finding the appropriate reaction duration (5-60 minutes). Acetylation was optimized at different steps of the process, including the initial volume of compounds to derivatize, the added amounts of Zn (0.1-0.25 g), the nature of the derivatization product (acetic anhydride, heptafluorobutyric acid…) and the liquid/liquid extraction at the end of the process. After derivatization, LODs were decreased by a factor 3 for OH-PAHs and by a factor 4 for quinones, all the quinones being now detected. Thereafter, quinones and OH-PAHs were extracted from spiked sediments using microwave assisted extraction (MAE) followed by GC-MS analysis. Several mixtures of solvents of different volumes (10-25 mL) and using different extraction temperatures (80-120°C) were tested to obtain the best recovery yields. Satisfactory recoveries could be obtained for quinones (70-96%) and for OH-PAHs (70-104%). Temperature was a critical factor which had to be controlled to avoid oxy-PAHs degradation during the MAE extraction process. Conclusion: Even if MAE-GC-MS was satisfactory to analyze these oxy-PAHs, MAE optimization has to be carried on to obtain a most appropriate extraction solvent mixture, allowing a direct injection in the HPLC-UV-FLD system, which is more sensitive than GC-MS and does not necessitate a previous long derivatization step.

Keywords: derivatizations for GC-MS, microwave assisted extraction, on-line HPLC-UV-FLD, oxygenated PAHs, polluted sediments

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74 Neuroprotection against N-Methyl-D-Aspartate-Induced Optic Nerve and Retinal Degeneration Changes by Philanthotoxin-343 to Alleviate Visual Impairments Involve Reduced Nitrosative Stress

Authors: Izuddin Fahmy Abu, Mohamad Haiqal Nizar Mohamad, Muhammad Fattah Fazel, Renu Agarwal, Igor Iezhitsa, Nor Salmah Bakar, Henrik Franzyk, Ian Mellor

Abstract:

Glaucoma is the global leading cause of irreversible blindness. Currently, the available treatment strategy only involves lowering intraocular pressure (IOP); however, the condition often progresses despite lowered or normal IOP in some patients. N-methyl-D-aspartate receptor (NMDAR) excitotoxicity often occurs in neurodegeneration-related glaucoma; thus it is a relevant target to develop a therapy based on neuroprotection approach. This study investigated the effects of Philanthotoxin-343 (PhTX-343), an NMDAR antagonist, on the neuroprotection of NMDA-induced glaucoma to alleviate visual impairments. Male Sprague-Dawley rats were equally divided: Groups 1 (control) and 2 (glaucoma) were intravitreally injected with phosphate buffer saline (PBS) and NMDA (160nM), respectively, while group 3 was pre-treated with PhTX-343 (160nM) 24 hours prior to NMDA injection. Seven days post-treatments, rats were subjected to visual behavior assessments and subsequently euthanized to harvest their retina and optic nerve tissues for histological analysis and determination of nitrosative stress level using 3-nitrotyrosine ELISA. Visual behavior assessments via open field, object, and color recognition tests demonstrated poor visual performance in glaucoma rats indicated by high exploratory behavior. PhTX-343 pre-treatment appeared to preserve visual abilities as all test results were significantly improved (p < 0.05). H&E staining of the retina showed a marked reduction of ganglion cell layer thickness in the glaucoma group; in contrast, PhTX-343 significantly increased the number by 1.28-folds (p < 0.05). PhTX-343 also increased the number of cell nuclei/100μm2 within inner retina by 1.82-folds compared to the glaucoma group (p < 0.05). Toluidine blue staining of optic nerve tissues showed that PhTX-343 reduced the degeneration changes compared to the glaucoma group which exhibited vacuolation overall sections. PhTX-343 also decreased retinal 3- nitrotyrosine concentration by 1.74-folds compared to the glaucoma group (p < 0.05). All results in PhTX-343 group were comparable to control (p > 0.05). We conclude that PhTX-343 protects against NMDA-induced changes and visual impairments in the rat model by reducing nitrosative stress levels.

Keywords: excitotoxicity, glaucoma, nitrosative stress , NMDA receptor , N-methyl-D-aspartate , philanthotoxin, visual behaviour

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73 Mechanical Properties of Carbon Fibre Reinforced Thermoplastic Composites Consisting of Recycled Carbon Fibres and Polyamide 6 Fibres

Authors: Mir Mohammad Badrul Hasan, Anwar Abdkader, Chokri Cherif

Abstract:

With the increasing demand and use of carbon fibre reinforced composites (CFRC), disposal of the carbon fibres (CF) and end of life composite parts is gaining tremendous importance on the issue especially of sustainability. Furthermore, a number of processes (e. g. pyrolysis, solvolysis, etc.) are available currently to obtain recycled CF (rCF) from end-of-life CFRC. Since the CF waste or rCF are neither allowed to be thermally degraded nor landfilled (EU Directive 1999/31/EC), profitable recycling and re-use concepts are urgently necessary. Currently, the market for materials based on rCF mainly consists of random mats (nonwoven) made from short fibres. The strengths of composites that can be achieved from injection-molded components and from nonwovens are between 200-404 MPa and are characterized by low performance and suitable for non-structural applications such as in aircraft and vehicle interiors. On the contrary, spinning rCF to yarn constructions offers good potential for higher CFRC material properties due to high fibre orientation and compaction of rCF. However, no investigation is reported till yet on the direct comparison of the mechanical properties of thermoplastic CFRC manufactured from virgin CF filament yarn and spun yarns from staple rCF. There is a lack of understanding on the level of performance of the composites that can be achieved from hybrid yarns consisting of rCF and PA6 fibres. In this drop back, extensive research works are being carried out at the Textile Machinery and High-Performance Material Technology (ITM) on the development of new thermoplastic CFRC from hybrid yarns consisting of rCF. For this purpose, a process chain is developed at the ITM starting from fibre preparation to hybrid yarns manufacturing consisting of staple rCF by mixing with thermoplastic fibres. The objective is to apply such hybrid yarns for the manufacturing of load bearing textile reinforced thermoplastic CFRCs. In this paper, the development of innovative multi-component core-sheath hybrid yarn structures consisting of staple rCF and polyamide 6 (PA 6) on a DREF-3000 friction spinning machine is reported. Furthermore, Unidirectional (UD) CFRCs are manufactured from the developed hybrid yarns, and the mechanical properties of the composites such as tensile and flexural properties are analyzed. The results show that the UD composite manufactured from the developed hybrid yarns consisting of staple rCF possesses approximately 80% of the tensile strength and E-module to those produced from virgin CF filament yarn. The results show a huge potential of the DREF-3000 friction spinning process to develop composites from rCF for high-performance applications.

Keywords: recycled carbon fibres, hybrid yarn, friction spinning, thermoplastic composite

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72 Properties of the CsPbBr₃ Quantum Dots Treated by O₃ Plasma for Integration in the Perovskite Solar Cell

Authors: Sh. Sousani, Z. Shadrokh, M. Hofbauerová, J. Kollár, M. Jergel, P. Nádaždy, M. Omastová, E. Majková

Abstract:

Perovskite quantum dots (PQDs) have the potential to increase the performance of the perovskite solar cell (PSCs). The integration of PQDs into PSCs can extend the absorption range and enhance photon harvesting and device efficiency. In addition, PQDs can stabilize the device structure by passivating surface defects and traps in the perovskite layer and enhance its stability. The integration of PQDs into PSCs is strongly affected by the type of ligands on the surface of PQDs. The ligands affect the charge transport properties of PQDs, as well as the formation of well-defined interfaces and stability of PSCs. In this work, the CsPbBr₃ QDs were synthesized by the conventional hot-injection method using cesium oleate, PbBr₂ and two different ligands, namely oleic acid (OA) oleylamine (OAm) and didodecyldimethylammonium bromide (DDAB). The STEM confirmed regular shape and relatively monodisperse cubic structure with an average size of about 10-14 nm of the prepared CsPbBr₃ QDs. Further, the photoluminescent (PL) properties of the PQDs/perovskite bilayer with the ligand OA, OAm and DDAB were studied. For this purpose, ITO/PQDs as well as ITO/PQDs/MAPI perovskite structures were prepared by spin coating and the effect of the ligand and oxygen plasma treatment was analyzed. The plasma treatment of the PQDs layer could be beneficial for the deposition of the MAPI perovskite layer and the formation of a well-defined PQDs/MAPI interface. The absorption edge in UV-Vis absorption spectra for OA, OAm CsPbBr₃ QDs is placed around 513 nm (the band gap 2.38 eV); for DDAB CsPbBr₃ QDs, it is located at 490 nm (the band gap 2.33 eV). The photoluminescence (PL) spectra of CsPbBr₃ QDs show two peaks located around 514 nm (503 nm) and 718 nm (708 nm) for OA, OAm (DDAB). The peak around 500 nm corresponds to the PL of PQDs, and the peak close to 710 nm belongs to the surface states of PQDs for both types of ligands. These surface states are strongly affected by the O₃ plasma treatment. For PQDs with DDAB ligand, the O₃ exposure (5, 10, 15 s) results in the blue shift of the PQDs peak and a non-monotonous change of the amplitude of the surface states' peak. For OA, OAm ligand, the O₃ exposition did not cause any shift of the PQDs peak, and the intensity of the PL peak related to the surface states is lower by one order of magnitude in comparison with DDAB, being affected by O₃ plasma treatment. The PL results indicate the possibility of tuning the position of the PL maximum by the ligand of the PQDs. Similar behavior of the PQDs layer was observed for the ITO/QDs/MAPI samples, where an additional strong PL peak at 770 nm coming from the perovskite layer was observed; for the sample with PQDs with DDAB ligands, a small blue shift of the perovskite PL maximum was observed independently of the plasma treatment. These results suggest the possibility of affecting the PL maximum position and the surface states of the PQDs by the combination of a suitable ligand and the O₃ plasma treatment.

Keywords: perovskite quantum dots, photoluminescence, O₃ plasma., Perovskite Solar Cells

Procedia PDF Downloads 65
71 Properties of the CsPbBr₃ Quantum Dots Treated by O₃ Plasma for Integration in the Perovskite Solar Cell

Authors: Sh. Sousani, Z. Shadrokh, M. Hofbauerová, J. Kollár, M. Jergel, P. Nádaždy, M. Omastová, E. Majková

Abstract:

Perovskite quantum dots (PQDs) have the potential to increase the performance of the perovskite solar cells (PSCs). The integration of PQDs into PSCs can extend the absorption range and enhance photon harvesting and device efficiency. In addition, PQDs can stabilize the device structure by passivating surface defects and traps in the perovskite layer and enhance its stability. The integration of PQDs into PSCs is strongly affected by the type of ligands on the surface of PQDs. The ligands affect the charge transport properties of PQDs, as well as the formation of well-defined interfaces and stability of PSCs. In this work, the CsPbBr₃ QDs were synthesized by the conventional hot-injection method using cesium oleate, PbBr₂, and two different ligands, namely oleic acid (OA)@oleylamine (OAm) and didodecyldimethylammonium bromide (DDAB). The STEM confirmed regular shape and relatively monodisperse cubic structure with an average size of about 10-14 nm of the prepared CsPbBr₃ QDs. Further, the photoluminescent (PL) properties of the PQDs/perovskite bilayer with the ligand OA@OAm and DDAB were studied. For this purpose, ITO/PQDs, as well as ITO/PQDs/MAPI perovskite structures, were prepared by spin coating, and the effect of the ligand and oxygen plasma treatment was analysed. The plasma treatment of the PQDs layer could be beneficial for the deposition of the MAPI perovskite layer and the formation of a well-defined PQDs/MAPI interface. The absorption edge in UV-Vis absorption spectra for OA@OAm CsPbBr₃ QDs is placed around 513 nm (the band gap 2.38 eV); for DDAB CsPbBr₃ QDs, it is located at 490 nm (the band gap 2.33 eV). The photoluminescence (PL) spectra of CsPbBr₃ QDs show two peaks located around 514 nm (503 nm) and 718 nm (708 nm) for OA@OAm (DDAB). The peak around 500 nm corresponds to the PL of PQDs, and the peak close to 710 nm belongs to the surface states of PQDs for both types of ligands. These surface states are strongly affected by the O₃ plasma treatment. For PQDs with DDAB ligand, the O₃ exposure (5, 10, 15 s) results in the blue shift of the PQDs peak and a non-monotonous change of the amplitude of the surface states' peak. For OA@OAm ligand, the O₃ exposition did not cause any shift of the PQDs peak, and the intensity of the PL peak related to the surface states is lower by one order of magnitude in comparison with DDAB, being affected by O₃ plasma treatment. The PL results indicate the possibility of tuning the position of the PL maximum by the ligand of the PQDs. Similar behaviour of the PQDs layer was observed for the ITO/QDs/MAPI samples, where an additional strong PL peak at 770 nm coming from the perovskite layer was observed; for the sample with PQDs with DDAB ligands, a small blue shift of the perovskite PL maximum was observed independently of the plasma treatment. These results suggest the possibility of affecting the PL maximum position and the surface states of the PQDs by the combination of a suitable ligand and the O₃ plasma treatment.

Keywords: perovskite quantum dots, photoluminescence, O₃ plasma., perovskite solar cells

Procedia PDF Downloads 71
70 Thermoplastic-Intensive Battery Trays for Optimum Electric Vehicle Battery Pack Performance

Authors: Dinesh Munjurulimana, Anil Tiwari, Tingwen Li, Carlos Pereira, Sreekanth Pannala, John Waters

Abstract:

With the rapid transition to electric vehicles (EVs) across the globe, car manufacturers are in need of integrated and lightweight solutions for the battery packs of these vehicles. An integral part of a battery pack is the battery tray, which constitutes a significant portion of the pack’s overall weight. Based on the functional requirements, cost targets, and packaging space available, a range of materials –from metals, composites, and plastics– are often used to develop these battery trays. This paper considers the design and development of integrated thermoplastic-intensive battery trays, using the available packaging space from a representative EV battery pack. Presented as a proposed alternative are multiple concepts to integrate several connected systems such as cooling plates and underbody impact protection parts of a multi-piece incumbent battery pack. The resulting digital prototype was evaluated for several mechanical performance measures such as mechanical shock, drop, crush resistance, modal analysis, and torsional stiffness. The performance of this alternative design is then compared with the incumbent solution. In addition, insights are gleaned into how these novel approaches can be optimized to meet or exceed the performance of incumbent designs. Preliminary manufacturing feasibility of the optimal solution using injection molding and other commonly used manufacturing methods for thermoplastics is briefly explained. Then numerical and analytical evaluations are performed to show a representative Pareto front of cost vs. volume of the production parts. The proposed solution is observed to offer weight savings of up to 40% on a component level and part elimination of up to two systems in the battery pack of a typical battery EV while offering the potential to meet the required performance measures highlighted above. These conceptual solutions are also observed to potentially offer secondary benefits such as improved thermal and electrical isolations and be able to achieve complex geometrical features, thus demonstrating the ability to use the complete packaging space available in the vehicle platform considered. The detailed study presented in this paper serves as a valuable reference for researches across the globe working on the development of EV battery packs – especially those with an interest in the potential of employing alternate solutions as part of a mixed-material system to help capture untapped opportunities to optimize performance and meet critical application requirements.

Keywords: thermoplastics, lightweighting, part integration, electric vehicle battery packs

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69 Direct Assessment of Cellular Immune Responses to Ovalbumin with a Secreted Luciferase Transgenic Reporter Mouse Strain IFNγ-Lucia

Authors: Martyna Chotomska, Aleksandra Studzinska, Marta Lisowska, Justyna Szubert, Aleksandra Tabis, Jacek Bania, Arkadiusz Miazek

Abstract:

Objectives: Assessing antigen-specific T cell responses is of utmost importance for the pre-clinical testing of prototype vaccines against intracellular pathogens and tumor antigens. Mainly two types of in vitro assays are used for this purpose 1) enzyme-linked immunospot (ELISpot) and 2) intracellular cytokine staining (ICS). Both are time-consuming, relatively expensive, and require manual dexterity. Here, we assess if a straightforward detection of luciferase activity in blood samples of transgenic reporter mice expressing a secreted Lucia luciferase under the transcriptional control of IFN-γ promoter parallels the sensitivity of IFNγ ELISpot assay. Methods: IFN-γ-LUCIA mouse strain carrying multiple copies of Lucia luciferase transgene under the transcriptional control of IFNγ minimal promoter were generated by pronuclear injection of linear DNA. The specificity of transgene expression and mobilization was assessed in vitro using transgenic splenocytes exposed to various mitogens. The IFN-γ-LUCIA mice were immunized with 50mg of ovalbumin (OVA) emulsified in incomplete Freund’s adjuvant three times every two weeks by subcutaneous injections. Blood samples were collected before and five days after each immunization. Luciferase activity was assessed in blood serum. Peripheral blood mononuclear cells were separated and assessed for frequencies of OVA-specific IFNγ-secreting T cells. Results: We show that in vitro cultured splenocytes of IFN-γ-LUCIA mice respond by 2 and 3 fold increase in secreted luciferase activity to T cell mitogens concanavalin A and phorbol myristate acetate, respectively but fail to respond to B cell-stimulating E.coli lipopolysaccharide. Immunization of IFN-γ-LUCIA mice with OVA leads to over 4 fold increase in luciferase activity in blood serum five days post-immunization with a barely detectable increase in OVA-specific, IFNγ-secreting T cells by ELISpot. Second and third immunizations, further increase the luciferase activity and coincidently also increase the frequencies of OVA-specific T cells by ELISpot. Conclusions: We conclude that minimally invasive monitoring of luciferase secretions in blood serum of IFN-γ-LUCIA mice constitutes a sensitive method for evaluating primary and memory Th1 responses to protein antigens. As such, this method may complement existing methods for rapid immunogenicity assessment of prototype vaccines.

Keywords: ELISpot, immunogenicity, interferon-gamma, reporter mice, vaccines

Procedia PDF Downloads 172