Search results for: beta-barrel membrane proteins
260 Epiphytic Growth on Filamentous Bacteria Found in Activated Sludge: A Morphological Approach
Authors: Thobela Conco, Sheena Kumari, Thor Stenstrom, Simona Rosetti, Valter Tandoi, Faizal Bux
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Filamentous bacteria are well documented as causative agents of bulking and foaming in the biological wastewater treatment process. These filamentous bacteria are however closely associated with other non-filamentous organism forming a micro-niche. Among these specific epiphytic bacteria attach to filaments in the consortium of organisms that make up the floc. Neither the eco-physiological role of the epiphytes nor the nature of the interaction between the epiphytic bacteria and the filament hosts they colonize is well understood and in need of in-depth investigations. The focus of this presentation is on the interaction between the epiphytic bacteria and the filament host. Samples from the activated sludge treatment have been repeatedly collected from several wastewater treatment plants in KwaZulu Natal. Extensive investigations have been performed with SEM and TEM electron microscopy, Polarized Light Microscopy with Congo red staining, and Thioflavin T staining to document the interaction. SEM was used to document the morphology of both the filament host and their epiphytes counterparts with the focus on the interface/point of contact between the two, while the main focus of the TEM investigations with the higher magnification aimed to document the ultra-structure features of two organisms relating to the interaction. The interaction of the perpendicular attachment partly seems to be governed by the physiological status of the filaments. The attachment further seems to trigger a response in the filaments with distinct internal visible structures at the attachment sites. It is postulated that these structures most likely are amyloid fibrils. Amyloid fibrils may play an overarching role in different types of attachments and has earlier been noted to play a significant role in biofilm formation in activated sludge. They also play a medical role in degenerative diseases such as Alzheimer’s and Diabetes. Further studies aims to define the eco-physiological role of amyloid fibrils in filamentous bacteria, based on their observed presence at interaction sites in this study. This will also relate to additional findings where selectivity within the species of epiphytes attaching to the selected filaments has been noted. The practical implications of the research findings is still to be determined, but the ecophysiological interaction between two closely associated species or groups may have significant impact in the future understanding of wastewater treatment processes and broaden existing knowledge on population dynamics.Keywords: activated sludge, amyloid proteins, epiphytic bacteria, filamentous bacteria
Procedia PDF Downloads 423259 Evaluation of Mito-Uncoupler Induced Hyper Metabolic and Aggressive Phenotype in Glioma Cells
Authors: Yogesh Rai, Saurabh Singh, Sanjay Pandey, Dhananjay K. Sah, B. G. Roy, B. S. Dwarakanath, Anant N. Bhatt
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One of the most common signatures of highly malignant gliomas is their capacity to metabolize more glucose to lactic acid than normal brain tissues, even under normoxic conditions (Warburg effect), indicating that aerobic glycolysis is constitutively upregulated through stable genetic or epigenetic changes. However, oxidative phosphorylation (OxPhos) is also required to maintain the mitochondrial membrane potential for tumor cell survival. In the process of tumorigenesis, tumor cells during fastest growth rate exhibit both high glycolytic and high OxPhos. Therefore, metabolically reprogrammed cancer cells with combination of both aerobic glycolysis and altered OxPhos develop a robust metabolic phenotype, which confers a selective growth advantage. In our study, we grew the high glycolytic BMG-1 (glioma) cells with continuous exposure of mitochondrial uncoupler 2, 4, dinitro phenol (DNP) for 10 passages to obtain a phenotype of high glycolysis with enhanced altered OxPhos. We found that OxPhos modified BMG (OPMBMG) cells has similar growth rate and cell cycle distribution but high mitochondrial mass and functional enzymatic activity than parental cells. In in-vitro studies, OPMBMG cells showed enhanced invasion, proliferation and migration properties. Moreover, it also showed enhanced angiogenesis in matrigel plug assay. Xenografted tumors from OPMBMG cells showed reduced latent period, faster growth rate and nearly five folds reduction in the tumor take in nude mice compared to BMG-1 cells, suggesting that robust metabolic phenotype facilitates tumor formation and growth. OPMBMG cells which were found radio-resistant, showed enhanced radio-sensitization by 2-DG as compared to the parental BMG-1 cells. This study suggests that metabolic reprogramming in cancer cells enhances the potential of migration, invasion and proliferation. It also strengthens the cancer cells to escape the death processes, conferring resistance to therapeutic modalities. Our data also suggest that combining metabolic inhibitors like 2-DG with conventional therapeutic modalities can sensitize such metabolically aggressive cancer cells more than the therapies alone.Keywords: 2-DG, BMG, DNP, OPM-BMG
Procedia PDF Downloads 221258 Alleviation of Adverse Effects of Salt Stress on Soybean (Glycine max. L.) by Using Osmoprotectants and Compost Application
Authors: Ayman El Sabagh, SobhySorour, AbdElhamid Omar, Adel Ragab, Mohammad Sohidul Islam, Celaleddin Barutçular, Akihiro Ueda, Hirofumi Saneoka
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Salinity is one of the major factors limiting crop production in an arid environment. What adds to the concern is that all the legume crops are sensitive to increasing soil salinity. So it is implacable to either search for salinity enhancement of legume plants. The exogenous of osmoprotectants has been found effective in reducing the adverse effects of salinity stress on plant growth. Despite its global importance soybean production suffer the problems of salinity stress causing damages at plant development. Therefore, in the current study we try to clarify the mechanism that might be involved in the ameliorating effects of osmo-protectants such as proline and glycine betaine and compost application on soybean plants grown under salinity stress. Experiments were carried out in the greenhouse of the experimental station, plant nutritional physiology, Hiroshima University, Japan in 2011- 2012. The experiment was arranged in a factorial design with 4 replications at NaCl concentrations (0 and 15 mM). The exogenous, proline and glycine betaine concentrations (0 mM and 25 mM) for each. Compost treatments (0 and 24 t ha-1). Results indicated that salinity stress induced reduction in all growth and physiological parameters (dry weights plant-1, chlorophyll content, N and K+ content) likewise, seed and quality traits of soybean plant compared with those of the unstressed plants. In contrast, salinity stress led to increases in the electrolyte leakage ratio, Na and proline contents. Thus tolerance against salt stress was observed, the improvement of salt tolerance resulted from proline, glycine betaine and compost were accompanied with improved membrane stability, K+, and proline accumulation on contrary, decreased Na+ content. These results clearly demonstrate that could be used to reduce the harmful effect of salinity on both physiological aspects and growth parameters of soybean. They are capable of restoring yield potential and quality of seed and may be useful in agronomic situations where saline conditions are diagnosed as a problem. Consequently, exogenous osmo-protectants combine with compost will effectively solve seasonal salinity stress problem and are a good strategy to increase salinity resistance in the drylands.Keywords: compost, glycine betaine, proline, salinity tolerance, soybean
Procedia PDF Downloads 364257 Hepatoprotective Action of Emblica officinalis Linn. against Radiation and Lead Induced Changes in Swiss Albino Mice
Authors: R. K. Purohit
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Ionizing radiation induces cellular damage through direct ionization of DNA and other cellular targets and indirectly via reactive oxygen species which may include effects from epigenetic changes. So there is a need of hour is to search for an ideal radioprotector which could minimize the deleterious and damaging effects caused by ionizing radiation. Radioprotectors are agents which reduce the radiation effects on cell when applied prior to exposure of radiation. The aim of this study was to access the efficacy of Emblica officinalis in reducing radiation and lead induced changes in mice liver. For the present experiment, healthy male Swiss albino mice (6-8 weeks) were selected and maintained under standard conditions of temperature and light. Fruit extract of Emblica was fed orally at the dose of 0.01 ml/animal/day. The animal were divided into seven groups according to the treatment i.e. lead acetate solution as drinking water (group-II) or exposed to 3.5 or 7.0 Gy gamma radiation (group-III) or combined treatment of radiation and lead acetate (group-IV). The animals of experimental groups were administered Emblica extract seven days prior to radiation or lead acetate treatment (group V, VI and VII) respectively. The animals from all the groups were sacrificed by cervical dislocation at each post-treatment intervals of 1, 2, 4, 7, 14 and 28 days. After sacrificing the animals pieces of liver were taken out and some of them were kept at -20°C for different biochemical parameters. The histopathological changes included cytoplasmic degranulation, vacuolation, hyperaemia, pycnotic and crenated nuclei. The changes observed in the control groups were compared with the respective experimental groups. An increase in the value of total proteins, glycogen, acid phosphtase, alkaline phosphatase activity and RNA was observed up to day-14 in the non drug treated group and day 7 in the Emblica treated groups, thereafter value declined up to day-28 without reaching to normal. The value of cholesterol and DNA showed a decreasing trend up to day -14 in non drug treated groups and day-7 in drug treated groups, thereafter value elevated up to day-28. The biochemical parameters were observed in the form of increase or decrease in the values. The changes were found dose dependent. After combined treatment of radiation and lead acetate synergistic effect were observed. The liver of Emblica treated animals exhibited less severe damage as compared to non-drug treated animals at all the corresponding intervals. An early and fast recovery was also noticed in Emblica pretreated animals. Thus, it appears that Emblica is potent enough to check lead and radiation induced heptic lesion in Swiss albino mice.Keywords: radiation, lead , emblica, mice, liver
Procedia PDF Downloads 313256 Structural Performance of Mechanically Connected Stone Panels under Cyclic Loading: Application to Aesthetic and Environmental Building Skin Design
Authors: Michel Soto Chalhoub
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Building designers in the Mediterranean region and other parts of the world utilize natural stone panels on the exterior façades as skin cover. This type of finishing is not only intended for aesthetic reasons but also environmental. The stone, since the earliest ages of civilization, has been used in construction and to-date some of the most appealing buildings owe their beauty to stone finishing. The stone also provides warmth in winter and freshness in summer as it moderates heat transfer and absorbs radiation. However, as structural codes became increasingly stringent about the dynamic performance of buildings, it became essential to study the performance of stone panels under cyclic loading – a condition that arises under the building is subjected to wind or earthquakes. The present paper studies the performance of stone panels using mechanical connectors when subjected to load reversal. In this paper, we present a theoretical model that addresses modes of failure in the steel connectors, by yield, and modes of failure in the stone, by fracture. Then we provide an experimental set-up and test results for rectangular stone panels of varying thickness. When the building is subjected to an earthquake, its rectangular panels within the structural system are subjected to shear deformations, which in turn impart stress into the stone cover. Rectangular stone panels, which typically range from 40cmx80cm to 60cmx120cm, need to be designed to withstand transverse loading from the direct application of lateral loads, and to withstand simultaneously in-plane loading (membrane stress) caused by inter-story drift and overall building lateral deflection. Results show correlation between the theoretical model which we derive from solid mechanics fundamentals and the experimental results, and lead to practical design recommendations. We find that for panel thickness below a certain threshold, it is more advantageous to utilize structural adhesive materials to connect stone panels to the main structural system of the building. For larger panel thicknesses, it is recommended to utilize mechanical connectors with special detailing to ensure a minimum level of ductility and energy dissipation.Keywords: solid mechanics, cyclic loading, mechanical connectors, natural stone, seismic, wind, building skin
Procedia PDF Downloads 254255 Assessment of Genetic Variability of Potato Genotypes for Proline Under Salt Stress Conditions
Authors: Elchin Hajiyev, Afet Memmedova Dadash, Sabina Hajiyeva, Aynur Karimova, Ramiz Aliyev
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Although potatoes have a wide distribution range, the yield potential of varieties varies greatly depending on the region. Our country is made up of agricultural regions with very different environmental characteristics.In this case, we cannot expect the introduced varieties to show the same adaptation to the different conditions of our country. For this reason, in our country, varieties with high general adaptability should be used, rather than varieties with special adaptability in certain areas. Soil salinization has become a global problem.Increased salinity has a serious impact on food security by reducing plant productivity. Plants have protective mechanisms of adaptation to salt stress, such as the synthesis of physiologically active substances, resistance to antioxidant stress and oxidation of membrane lipids. One of these substances is free proline. Our study revealed genetic variation in proline accumulation among samples exposed to stress factors.Changes in proline content under stress conditions were studied in 50 samples. There was wide variation across all treatments.The amount of proline varied between 7.2–37.7 μM/g under salinity conditions.The lowest rate was in the SF33 genotype (1.5 times more than the control (2.5 μM/g)).The highest level of proline under the influence of salt stress was in the SF45 genotype (7.25 times higher than the control (32.5 μM/g)). Our studies have found that the protective system reacts differently to the influence of stress factors. According to the results obtained on the amount of proline, adaptation mechanisms must be more actively activated to maintain metabolism and ensure viability in sensitive forms under the influence of stress factors. At high doses of the salt stressor, a tenfold increase in proline compared to the control indicates significant damage to the plant organism as a result of stress.To prevent damage to the body, the antioxidant system needs to quickly mobilize and work at full capacity in adverse conditions. An increase in the dose of the stress factor salt in our study caused a greater increase in the amount of free proline in plant tissues. Considering the functions of proline as an osmoprotector and antioxidant, it was found that increasing its amount is aimed at protecting the plant from the acute effects of stressors.Keywords: genetic variability, potato, genotypes, proline, stress
Procedia PDF Downloads 37254 The Effect of SIAH1 on PINK1 Homeostasis in Parkinson Disease
Authors: Fatimah Abd Elghani, Raymonde Szargel, Vered Shani, Hazem Safory, Haya Hamza, Mor Savyon, Ruth Rott, Rina Bandopadhyay, Simone Engelender
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Background: PINK1 is a mitochondrial kinase mutated in some familial cases of Parkinson’s disease. Down regulation of PINK1 results in abnormal mitochondrial morphology and altered membrane potential. Although PINK1 has a predicted mitochondrial import sequence, it’s cellular, and submitochondrial localization remains unclear, in part because it is rapidly degraded. In this work, we investigated the mechanisms involved in PINK1 degradation and how this may affect PINK1 stability and function, with implications for mitochondrial function in PD. In addition, pharmacological inhibition of proteasome activity was shown to lead to PINK1 accumulation, indicating that PINK1 degradation depends on the ubiquitin-proteasome system (UPS). Methods: Using co-immunoprecipitation assays, we identified E3 ubiquitin ligase SIAH1 as a PINK1-interacting protein in HEK293 cells as well as on rat brain tissues. In addition, we determined the effect of SIAH 1, SIAH2 and Parkin on PINK1 steady-state levels by Western blot analysis, and checked their possibility to ubiquitinate and mediate PINK1 degradation through the proteasome carried out in vivo ubiquitination experiments. Results: We have obtained results showing that SIAH-1 interacts with and ubiquitinates PINK1. The ubiquitination promoted by SIAH-1 leads to the proteasomal degradation of PINK1. We confirmed these findings by knocking down SIAH-1 and observing important accumulation of PINK1 in cells. Besides, we found that SIAH-1 decreases PINK1 steady-state levels but not the E3 ligase Parkin. We also investigated the interaction of SIAH-1 with PINK1 disease mutants and its ability to promote their ubiquitination and degradation. Although, no clear difference in the ability of SIAH-1 to promote the degradation of PINK1 disease mutants was observed. It is possible that dysfunction of proteasomal activity in the disease may lead to the accumulation and aggregation of ubiquitinated PINK1 in patients with PINK1 mutations, with possible implications to the pathogenesis of PD. Conclusions: Here, we demonstrated that SIAH-1 ubiquitinates and promotes the degradation of PINK1. In addition, SIAH-1 represents now a target that may help the improvement of mitophagy in PD. Further investigations needed to understand how mitophagy is regulated by PINK1-SIAH-1 axis to provide targets for future therapeutics.Keywords: PD, Parkinson's disease, PINK1, PTEN-induced kinase1, SIAH, seven in absentia homolog, SN, substantia nigra
Procedia PDF Downloads 136253 Extracorporeal Co2 Removal (Ecco2r): An Option for Treatment for Refractory Hypercapnic Respiratory Failure
Authors: Shweh Fern Loo, Jun Yin Ong, Than Zaw Oo
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Acute respiratory distress syndrome (ARDS) is a common serious condition of bilateral lung infiltrates that develops secondary to various underlying conditions such as diseases or injuries. ARDS with severe hypercapnia is associated with higher ICU mortality and morbidity. Venovenous Extracorporeal membrane oxygenation (VV-ECMO) support has been established to avert life-threatening hypoxemia and hypercapnic respiratory failure despite optimal conventional mechanical ventilation. However, VV-ECMO is relatively not advisable in particular groups of patients, especially in multi-organ failure, advanced age, hemorrhagic complications and irreversible central nervous system pathology. We presented a case of a 79-year-old Chinese lady without any pre-existing lung disease admitted to our hospital intensive care unit (ICU) after acute presentation of breathlessness and chest pain. After extensive workup, she was diagnosed with rapidly progressing acute interstitial pneumonia with ARDS and hypercapnia respiratory failure. The patient received lung protective strategies of mechanical ventilation and neuromuscular blockage therapy as per clinical guidelines. However, hypercapnia respiratory failure was refractory, and she was deemed not a good candidate for VV-ECMO support given her advanced age and high vasopressor requirements from shock. Alternative therapy with extracorporeal CO2 removal (ECCO2R) was considered and implemented. The patient received 12 days of ECCO2R paired with muscle paralysis, optimization of lung-protective mechanical ventilation and dialysis. Unfortunately, the patient still had refractory hypercapnic respiratory failure with dual vasopressor support despite prolonged therapy. Given failed and futile medical treatment, the family opted for withdrawal of care, a conservative approach, and comfort care, which led to her demise. The effectivity of extracorporeal CO2 removal may depend on disease burden, involvement and severity of the disease. There is insufficient data to make strong recommendations about its benefit-risk ratio for ECCO2R devices, and further studies and data would be required. Nonetheless, ECCO2R can be considered an alternative treatment for refractory hypercapnic respiratory failure patients who are unsuitable for initiating venovenous ECMO.Keywords: extracorporeal CO2 removal (ECCO2R), acute respiratory distress syndrome (ARDS), acute interstitial pneumonia (AIP), hypercapnic respiratory failure
Procedia PDF Downloads 60252 Acerola and Orange By-Products as Sources of Bioactive Compounds for Probiotic Fermented Milks
Authors: Tatyane Lopes de Freitas, Antonio Diogo S. Vieira, Susana Marta Isay Saad, Maria Ines Genovese
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The fruit processing industries generate a large volume of residues to produce juices, pulps, and jams. These residues, or by-products, consisting of peels, seeds, and pulps, are routinely discarded. Fruits are rich in bioactive compounds, including polyphenols, which have positive effects on health. Dry residues from two fruits, acerola (M. emarginata D. C.) and orange (C. sinensis), were characterized in relation to contents of ascorbic acid, minerals, total dietary fibers, moisture, ash, lipids, proteins, and carbohydrates, and also high performance liquid chromatographic profile of flavonoids, total polyphenols and proanthocyanidins contents, and antioxidant capacity by three different methods (Ferric reducing antioxidant power assay-FRAP, Oxygen Radical Absorbance Capacity-ORAC, 1,1-diphenyl-2-picrylhydrazil (DPPH) radical scavenging activity). Acerola by-products presented the highest acid ascorbic content (605 mg/100 g), and better antioxidant capacity than orange by-products. The dry residues from acerola demonstrated high contents of proanthocyanidins (617 µg CE/g) and total polyphenols (2525 mg gallic acid equivalents - GAE/100 g). Both presented high total dietary fiber (above 60%) and protein contents (acerola: 10.4%; orange: 9.9%), and reduced fat content (acerola: 1.6%; orange: 2.6%). Both residues showed high levels of potassium, calcium, and magnesium, and were considered sources of these minerals. With acerola by-product, four formulations of probiotics fermented milks were produced: F0 (without the addition of acerola residue (AR)), F2 (2% AR), F5 (5% AR) and F10 (10% AR). The physicochemical characteristics of the fermented milks throughout of storage were investigated, as well as the impact of in vitro simulated gastrointestinal conditions on flavonoids and probiotics. The microorganisms analyzed maintained their populations around 8 log CFU/g during storage. After the gastric phase of the simulated digestion, the populations decreased, and after the enteric phase, no colonies were detected. On the other hand, the flavonoids increased after the gastric phase, maintaining or suffering small decrease after enteric phase. Acerola by-products powder is a valuable ingredient to be used in functional foods because is rich in vitamin C, fibers and flavonoids. These flavonoids appear to be highly resistant to the acids and salts of digestion.Keywords: acerola, orange, by-products, fermented milk
Procedia PDF Downloads 127251 Protective Role of Curcumin against Ionising Radiation of Gamma Ray
Authors: Turban Kar, Maitree Bhattacharyya
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Curcumin, a dietary antioxidant has been identified as a wonder molecule to possess therapeutic properties protecting the cellular macromolecules from oxidative damage. In our experimental study, we have explored the effectiveness of curcumin in protecting the structural paradigm of Human Serum Albumin (HSA) when exposed to gamma irradiation. HSA, being an important transport protein of the circulatory system, is involved in binding of variety of metabolites, drugs, dyes and fatty acids due to the presence of hydrophobic pockets inside the structure. HSA is also actively involved in the transportation of drugs and metabolites to their targets, because of its long half-life and regulation of osmotic blood pressure. Gamma rays, in its increasing concentration, results in structural alteration of the protein and superoxide radical generation. Curcumin, on the other hand, mitigates the damage, which has been evidenced in the following experiments. Our study explores the possibility for protection by curcumin during the molecular and conformational changes of HSA when exposed to gamma irradiation. We used a combination of spectroscopic methods to probe the conformational ensemble of the irradiated HSA and finally evaluated the extent of restoration by curcumin. SDS - PAGE indicated the formation of cross linked aggregates as a consequence of increasing exposure of gamma radiation. CD and FTIR spectroscopy inferred significant decrease in alpha helix content of HSA from 57% to 15% with increasing radiation doses. Steady state and time resolved fluorescence studies complemented the spectroscopic measurements when lifetime decay was significantly reduced from 6.35 ns to 0.37 ns. Hydrophobic and bityrosine study showed the effectiveness of curcumin for protection against radiation induced free radical generation. Moreover, bityrosine and hydrophobic profiling of gamma irradiated HSA in presence and absence of curcumin provided light on the formation of ROS species generation and the protective (magical) role of curcumin. The molecular mechanism of curcumin protection to HSA from gamma irradiation is yet unknown, though a possible explanation has been proposed in this work using Thioflavin T assay. It was elucidated, that when HSA is irradiated at low dose of gamma radiation in presence of curcumin, it is capable of retaining the native characteristic properties to a greater extent indicating stabilization of molecular structure. Thus, curcumin may be utilized as a therapeutic strategy to protect cellular proteins.Keywords: Bityrosine content, conformational change, curcumin, gamma radiation, human serum albumin
Procedia PDF Downloads 150250 Anti-Acanthamoeba Activities of Fatty Acid Salts and Fatty Acids
Authors: Manami Masuda, Mariko Era, Takayoshi Kawahara, Takahide Kanyama, Hiroshi Morita
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Objectives: Fatty acid salts are a type of anionic surfactant and are produced from fatty acids and alkali. Moreover, fatty acid salts are known to have potent antibacterial activities. Acanthamoeba is ubiquitously distributed in the environment including sea water, fresh water, soil and even from the air. Although generally free-living, Acanthamoeba can be an opportunistic pathogen, which could cause a potentially blinding corneal infection known as Acanthamoeba keratitis. So, in this study, we evaluated the anti-amoeba activity of fatty acid salts and fatty acids to Acanthamoeba castellanii ATCC 30010. Materials and Methods: The antibacterial activity of 9 fatty acid salts (potassium butyrate (C4K), caproate (C6K), caprylate (C8K), caprate (C10K), laurate (C12K), myristate (C14K), oleate (C18:1K), linoleate (C18:2K), linolenate (C18:3K)) tested on cells of Acanthamoeba castellanii ATCC 30010. Fatty acid salts (concentration of 175 mM and pH 10.5) were prepared by mixing the fatty acid with the appropriate amount of KOH. The amoeba suspension mixed with KOH with a pH adjusted solution was used as the control. Fatty acids (concentration of 175 mM) were prepared by mixing the fatty acid with Tween 80 (20 %). The amoeba suspension mixed with Tween 80 (20 %) was used as the control. The anti-amoeba method, the amoeba suspension (3.0 × 104 cells/ml trophozoites) was mixed with the sample of fatty acid potassium (final concentration of 175 mM). Samples were incubated at 30°C, for 10 min, 60 min, and 180 min and then the viability of A. castellanii was evaluated using plankton counting chamber and trypan blue stainings. The minimum inhibitory concentration (MIC) against Acanthamoeba was determined using the two-fold dilution method. The MIC was defined as the minimal anti-amoeba concentration that inhibited visible amoeba growth following incubation (180 min). Results: C8K, C10K, and C12K were the anti-amoeba effect of 4 log-unit (99.99 % growth suppression of A. castellanii) incubated time for 180 min against A. castellanii at 175mM. After the amoeba, the suspension was mixed with C10K or C12K, destroying the cell membrane had been observed. Whereas, the pH adjusted control solution did not exhibit any effect even after 180 min of incubation with A. castellanii. Moreover, C6, C8, and C18:3 were the anti-amoeba effect of 4 log-unit incubated time for 60 min. C4 and C18:2 exhibited a 4-log reduction after 180 min incubation. Furthermore, the minimum inhibitory concentration (MIC) was determined. The MIC of C10K, C12K and C4 were 2.7 mM. These results indicate that C10K, C12K and C4 have high anti-amoeba activity against A. castellanii and suggest C10K, C12K and C4 have great potential for antimi-amoeba agents.Keywords: Fatty acid salts, anti-amoeba activities, Acanthamoeba, fatty acids
Procedia PDF Downloads 472249 Effects of Nanoencapsulated Echinacea purpurea Ethanol Extract on the Male Reproductive Function in Streptozotocin-Induced Diabetic Rats
Authors: Jia-Ling Ho, Xiu-Ru Zhang, Zwe-Ling Kong
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Diabetes mellitus (DM) is a major health problem that affects patients’ life quality throughout the world due to its many complications. It characterized by chronic hyperglycemia with oxidative stress, which impaired male reproductive function. Fibroblast growth factor 21 (FGF21) is a metabolic regulator that is required for normal spermatogenesis and protects against diabetes-induced germ cell apoptosis. Echinacea purpurea ethanol extract (EE), which contain phenolic acid and isobutylamide, had been proven to have antidiabetic property. Silica-chitosan nanoparticles (Nano-CS) has drug delivery and controlled release properties. This study aims to investigate whether silica-chitosan nanoparticles encapsulated EE (Nano-EE) had more ameliorating male infertility by analyzing the effect of testicular FGF21. The Nano-EE was characterized before used to treatment the diabetic rat model. Male Sprague-Dawley (SD) rats were obtained and divided into seven groups. A group was no induced Streptozotocin (STZ), marked as normal group. Diabetic rats were induced into diabetes by STZ (33 mg/kg). A diabetic group was no treatment with sample (diabetic control group), and other groups were treatment by Nano-CS (465 mg/kg), Nano-EE (93, 279, 465 mg/kg), and metformin (Met) (200 mg/kg) used as reference drug for 7 weeks. Our results indicated that the average nanoparticle size and zeta potential of Nano-EE were 2630 nm and -21.3 mV, respectively. The encapsulation ratio of Nano-EE was about 70%. It also confirmed the antioxidative activity was unchanged by comparing the DPPH and ABTS scavenging of Nano-EE and EE. In vivo test, Nano-EE can improve the STZ induced hyperglycemia, insulin resistance, and plasma FGF21 levels. Nano-EE has increased sperm motility, mitochondria membrane potential (MMP), plasma testosterone level, and reduction of abnormal sperm, nitric oxide (NO), superoxide production as well as reactive oxygen species (ROS). In addition, in plasma antioxidant enzymes glutathione peroxidase (GPx) and superoxide dismutase (SOD) was increased whereas pro-inflammatory cytokines TNF-α, and IL-1β were decreased. Further, in testis, protein content of FGF21, PGC-1α, and SIRT1 were improved. Nano-EE might improve diabetes-induced down-regulation of testicular FGF21 and SIRT1/PGC-1α signaling hence maintain spermatogenesis.Keywords: diabetes mellitus, Echinacea purpurea, reproductive dysfunction, silica-chitosan nanoparticles
Procedia PDF Downloads 186248 The Toxicity of Doxorubicin Connected with Nanotransporters
Authors: Iva Blazkova, Amitava Moulick, Vedran Milosavljevic, Pavel Kopel, Marketa Vaculovicova, Vojtech Adam, Rene Kizek
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Doxorubicin is one of the most commonly used and the most effective chemotherapeutic drugs. This antracycline drug isolated from the bacteria Streptomyces peuceticus var. caesius is sold under the trade name Adriamycin (hydroxydaunomycin, hydroxydaunorubicin). Doxorubicin is used in single therapy to treat hematological malignancies (blood cancers, leukaemia, lymphoma), many types of carcinoma (solid tumors) and soft tissue sarcomas. It has many serious side effects like nausea and vomiting, hair lost, myelosupression, oral mucositis, skin reactions and redness, but the most serious one is the cardiotoxicity. Because of the risk of heart attack and congestive heart failure, the total dose administered to patients has to be accurately monitored. With the aim to lower the side effects and to targeted delivery of doxorubicin into the tumor tissue, the different nanoparticles are studied. The drug can be bound on a surface of nanoparticle, encapsulated in the inner cavity, or incorporated into the structure of nanoparticle. Among others, carbon nanoparticles (graphene, carbon nanotubes, fullerenes) are highly studied. Besides the number of inorganic nanoparticles, a great potential exhibit also organic ones mainly lipid-based and polymeric nanoparticle. The aim of this work was to perform a toxicity study of free doxorubicin compared to doxorubicin conjugated with various nanotransporters. The effect of liposomes, fullerenes, graphene, and carbon nanotubes on the toxicity was analyzed. As a first step, the binding efficacy of between doxorubicin and the nanotransporter was determined. The highest efficacy was detected in case of liposomes (85% of applied drug was encapsulated) followed by graphene, carbon nanotubes and fullerenes. For the toxicological studies, the chicken embryos incubated under controlled conditions (37.5 °C, 45% rH, rotation every 2 hours) were used. In 7th developmental day of chicken embryos doxorubicin or doxorubicin-nanotransporter complex was applied on the chorioallantoic membrane of the eggs and the viability was analyzed every day till the 17th developmental day. Then the embryos were extracted from the shell and the distribution of doxorubicin in the body was analyzed by measurement of organs extracts using laser induce fluorescence detection. The chicken embryo mortality caused by free doxorubicin (30%) was significantly lowered by using the conjugation with nanomaterials. The highest accumulation of doxorubicin and doxorubicin nanotransporter complexes was observed in the liver tissueKeywords: doxorubicin, chicken embryos, nanotransporters, toxicity
Procedia PDF Downloads 445247 Small Community’s Proactive Thinking to Move from Zero to 100 Percent Water Reuse
Authors: Raj Chavan
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The City of Jal serves a population of approximately 3,500 people, including 2,100 permanent inhabitants and 1,400 oil and gas sector workers and RV park occupants. Over the past three years, Jal's population has increased by about 70 percent, mostly due to the oil and gas industry. The City anticipates that the population will exceed 4,200 by 2020, necessitating the construction of a new wastewater treatment plant (WWTP) because the old plant (aerated lagoon system) cannot accommodate such rapid population expansion without major renovations or replacement. Adhering to discharge permit restrictions has been challenging due to aging infrastructure and equipment replacement needs, as well as increasing nutrient loading to the wastewater collecting system from the additional oil and gas residents' recreational vehicles. The WWTP has not been able to maintain permit discharge standards for total nitrogen of less than 20 mg N/L and other characteristics in recent years. Based on discussions with the state's environmental department, it is likely that the future permit renewal would impose stricter conditions. Given its location in the dry, western part of the country, the City must rely on its meager groundwater supplies and scant annual precipitation. The city's groundwater supplies will be depleted sooner than predicted due to rising demand from the growing population for drinking, leisure, and other industrial uses (fracking). The sole type of reuse the city was engaging in (recreational reuse for a golf course) had to be put on hold because of an effluent water compliance issue. As of right now, all treated effluent is evaporated. The city's long-term goal is to become a zero-waste community that sends all of its treated wastewater effluent either to the golf course, Jal Lake, or the oil and gas industry for reuse. Hydraulic fracturing uses a lot of water, but if the oil and gas industry can use recycled water, it can reduce its impact on freshwater supplies. The City's goal of 100% reuse has been delayed by the difficulties of meeting the constraints of the regular discharge permit due to the large rise in influent loads and the aging infrastructure. The City of Jal plans to build a new WWTP that can keep up with the city's rapid population increase due to the oil and gas industry. Several treatment methods were considered in light of the City's needs and its long-term goals, but MBR was ultimately chosen recommended since it meets all of the permit's requirements while also providing 100 percent beneficial reuse. This talk will lay out the plan for the city to reach its goal of 100 percent reuse, as well as the various avenues for funding the small community that have been considered.Keywords: membrane bioreactor, nitrogent, reuse, small community
Procedia PDF Downloads 82246 Rhizobia-Containing Rhizobacterial Consortia and Intercropping Improved Faba Bean and Wheat Performances Under Stress Combining Drought and Phosphorus Deficiency
Authors: Said Cheto, Khawla Oukaltouma, Imane Chamkhi, Ammar Ibn Yasser, Bouchra Benmrid, Ahmed Qaddoury, Lamfeddal Kouisni, Joerg Geistlinger, Youssef Zeroual, Adnane Bargaz, Cherki Ghoulam
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Our study aimed to assess, the role of inoculation of faba bean/wheat intercrops with selected rhizobacteria consortia gathering one rhizobia and two phosphate solubilizing bacteria “PSB” to alleviate the effects of combined water deficit and P limitation on Faba bean/ wheat intercrops versus monocrops under greenhouse conditions. One Vicia faba L variety (Aguadulce “Ag”), and one Triticum durum L. variety (Karim “K”) were grown as sole crops or intercrop in pots containing sterilized substrate (sand: peat 4:1v/v) added either with rock phosphate (RP) as the alone P source (P limitation) or with KH₂PO₄ in nutrient solution (P sufficient control). Plant inoculation was done using rhizobacterial consortia composed; C1(Rhizobium laguerreae, Kocuria sp, and Pseudomonas sp) and C2 (R. laguerreae, Rahnella sp, and Kocuria sp). Two weeks after inoculation, the plants were submitted to water deficit consisting of 40% of substrate water holding Capacity (WHC) versus 80% WHC for well-watered plants. At the flowering stage, the trial was assessed, and the results showed that inoculation with both consortia (C1 and C2) improved faba bean biomass in terms of shoots, roots, and nodules compared to inoculation with rhizobia alone, particularly C2 improved these parametres by 19.03, 78.99, and 72.73%, respectively. Leaf relative water content decreased under combined stress, particularly in response to C1 with a significant improvement of this parameter in wheat intercrops. For faba bean under P limitation, inoculation with C2 increased stomatal conductance (gs) by 35.73% compared to plants inoculated with rhizobia alone. Furthermore, the same inoculum C2 improved membrane stability by 44,33% versus 16,16% for C1 compared to inoculation with rhizobia alone under P deficit. For sole cropped faba bean plants, inoculation with both consortia improved N accumulation compared to inoculation with rhizobia alone with an increase of 70.75% under P limitation. Moreover, under the combined stress, intercropping inoculation with C2 improved plant biomass and N content (112.98%) in wheat plants, compared to the sole crop. Our finding revealed that consortium C2 might offer an agronomic advantage under water and P deficit and could be used as inoculum for enhancing faba bean and wheat production under both monocropping and intercropping systems.Keywords: drought, phosphorus, intercropping, PSB, rhizobia, vicia faba, Triticum durum
Procedia PDF Downloads 67245 Transdermal Delivery of Sodium Diclofenac from Palm Kernel Oil Esteres Nanoemulsions
Authors: Malahat Rezaee, Mahiran Basri, Abu Bakar Salleh, Raja Noor Zaliha Raja Abdul Rahman
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Sodium diclofenac is one of the most commonly used drugs of nonsteroidal anti-inflammatory drugs (NSAIDs). It is especially effective in the controlling the severe conditions of inflammation and pain, musculoskeletal disorders, arthritis, and dysmenorrhea. Formulation as nanoemulsions is one of the nanoscience approaches that has been progressively considered in pharmaceutical science for transdermal delivery of the drug. Nanoemulsions are a type of emulsion with particle sizes ranging from 20 nm to 200 nm. An emulsion is formed by the dispersion of one liquid, usually the oil phase in another immiscible liquid, water phase that is stabilized using the surfactant. Palm kernel oil esters (PKOEs), in comparison to other oils, contain higher amounts of shorter chain esters, which suitable to be applied in micro and nanoemulsion systems as a carrier for actives, with excellent wetting behavior without the oily feeling. This research aimed to study the effect of terpene type and concentration on sodium diclofenac permeation from palm kernel oil esters nanoemulsions and physicochemical properties of the nanoemulsions systems. The effect of various terpenes of geraniol, menthone, menthol, cineol and nerolidol at different concentrations of 0.5, 1.0, 2.0, and 4.0% on permeation of sodium diclofenac were evaluated using Franz diffusion cells and rat skin as permeation membrane. The results of this part demonstrated that all terpenes showed promoting effect on sodium diclofenac penetration. However, menthol and menthone at all concentrations showed significant effects (<0.05) on drug permeation. The most outstanding terpene was menthol with the most significant effect for skin permeability of sodium diclofenac. The effect of terpenes on physicochemical properties of nanoemulsion systems was investigated on the parameters of particle size, zeta potential, pH, viscosity and electrical conductivity. The result showed that all terpenes had the significant effect on particle size and non-significant effects on the zeta potential of the nanoemulsion systems. The effect of terpenes was significant on pH, excluding the menthone at concentrations of 0.5 and 1.0%, and cineol and nerolidol at the concentration of 2.0%. Terpenes also had significant effect on viscosity of nanoemulsions exception of menthone and cineol at the concentration of 0.5%. The result of conductivity measurements showed that all terpenes at all concentration except cineol at the concentration of 0.5% represented significant effect on electrical conductivity.Keywords: nanoemulsions, palm kernel oil esters, sodium diclofenac, terpenes, skin permeation
Procedia PDF Downloads 414244 Prevalence of Breast Cancer Molecular Subtypes at a Tertiary Cancer Institute
Authors: Nahush Modak, Meena Pangarkar, Anand Pathak, Ankita Tamhane
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Background: Breast cancer is the prominent cause of cancer and mortality among women. This study was done to show the statistical analysis of a cohort of over 250 patients detected with breast cancer diagnosed by oncologists using Immunohistochemistry (IHC). IHC was performed by using ER; PR; HER2; Ki-67 antibodies. Materials and methods: Formalin fixed Paraffin embedded tissue samples were obtained by surgical manner and standard protocol was followed for fixation, grossing, tissue processing, embedding, cutting and IHC. The Ventana Benchmark XT machine was used for automated IHC of the samples. Antibodies used were supplied by F. Hoffmann-La Roche Ltd. Statistical analysis was performed by using SPSS for windows. Statistical tests performed were chi-squared test and Correlation tests with p<.01. The raw data was collected and provided by National Cancer Insitute, Jamtha, India. Result: Luminal B was the most prevailing molecular subtype of Breast cancer at our institute. Chi squared test of homogeneity was performed to find equality in distribution and Luminal B was the most prevalent molecular subtype. The worse prognostic indicator for breast cancer depends upon expression of Ki-67 and her2 protein in cancerous cells. Our study was done at p <.01 and significant dependence was observed. There exists no dependence of age on molecular subtype of breast cancer. Similarly, age is an independent variable while considering Ki-67 expression. Chi square test performed on Human epidermal growth factor receptor 2 (HER2) statuses of patients and strong dependence was observed in percentage of Ki-67 expression and Her2 (+/-) character which shows that, value of Ki depends upon Her2 expression in cancerous cells (p<.01). Surprisingly, dependence was observed in case of Ki-67 and Pr, at p <.01. This shows that Progesterone receptor proteins (PR) are over-expressed when there is an elevation in expression of Ki-67 protein. Conclusion: We conclude from that Luminal B is the most prevalent molecular subtype at National Cancer Institute, Jamtha, India. There was found no significant correlation between age and Ki-67 expression in any molecular subtype. And no dependence or correlation exists between patients’ age and molecular subtype. We also found that, when the diagnosis is Luminal A, out of the cohort of 257 patients, no patient shows >14% Ki-67 value. Statistically, extremely significant values were observed for dependence of PR+Her2- and PR-Her2+ scores on Ki-67 expression. (p<.01). Her2 is an important prognostic factor in breast cancer. Chi squared test for Her2 and Ki-67 shows that the expression of Ki depends upon Her2 statuses. Moreover, Ki-67 cannot be used as a standalone prognostic factor for determining breast cancer.Keywords: breast cancer molecular subtypes , correlation, immunohistochemistry, Ki-67 and HR, statistical analysis
Procedia PDF Downloads 117243 Triazenes: Unearthing Their Hidden Arsenal Against Malaria and Microbial Menace
Authors: Frans J. Smit, Wisdom A. Munzeiwa, Hermanus C. M. Vosloo, Lyn-Marie Birkholtz, Richard K. Haynes
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Malaria and antimicrobial infections remain significant global health concerns, necessitating the continuous search for novel therapeutic approaches. This abstract presents an overview of the potential use of triazenes as effective agents against malaria and various antimicrobial pathogens. Triazenes are a class of compounds characterized by a linear arrangement of three nitrogen atoms, rendering them structurally distinct from their cyclic counterparts. This study investigates the efficacy of triazenes against malaria and explores their antimicrobial activity. Preliminary results revealed significant antimalarial activity of the triazenes, as evidenced by in vitro screening against P. falciparum, the causative agent of malaria. Furthermore, the compounds exhibited broad-spectrum antimicrobial activity, indicating their potential as effective antimicrobial agents. These compounds have shown inhibitory effects on various essential enzymes and processes involved in parasite survival, replication, and transmission. The mechanism of action of triazenes against malaria involves interactions with critical molecular targets, such as enzymes involved in the parasite's metabolic pathways and proteins responsible for host cell invasion. The antimicrobial activity of the triazenes against bacteria and fungi was investigated through disc diffusion screening. The antimicrobial efficacy of triazenes has been observed against both Gram-positive and Gram-negative bacteria, as well as multidrug-resistant strains, making them potential candidates for combating drug-resistant infections. Furthermore, triazenes possess favourable physicochemical properties, such as good stability, solubility, and low toxicity, which are essential for drug development. The structural versatility of triazenes allows for the modification of their chemical composition to enhance their potency, selectivity, and pharmacokinetic properties. These modifications can be tailored to target specific pathogens, increasing the potential for personalized treatment strategies. In conclusion, this study highlights the potential of triazenes as promising candidates for the development of novel antimalarial and antimicrobial therapeutics. Further investigations are necessary to determine the structure-activity relationships and optimize the pharmacological properties of these compounds. The results warrant additional research, including MIC studies, to further explore the antimicrobial activity of the triazenes. Ultimately, these findings contribute to the development of more effective strategies for combating malaria and microbial infections.Keywords: malaria, anti-microbials, triazene, resistance
Procedia PDF Downloads 94242 Component Test of Martensitic/Ferritic Steels and Nickel-Based Alloys and Their Welded Joints under Creep and Thermo-Mechanical Fatigue Loading
Authors: Daniel Osorio, Andreas Klenk, Stefan Weihe, Andreas Kopp, Frank Rödiger
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Future power plants currently face high design requirements due to worsening climate change and environmental restrictions, which demand high operational flexibility, superior thermal performance, minimal emissions, and higher cyclic capability. The aim of the paper is, therefore, to investigate the creep and thermo-mechanical material behavior of improved materials experimentally and welded joints at component scale under near-to-service operating conditions, which are promising for application in highly efficient and flexible future power plants. These materials promise an increase in flexibility and a reduction in manufacturing costs by providing enhanced creep strength and, therefore, the possibility for wall thickness reduction. At the temperature range between 550°C and 625°C, the investigation focuses on the in-phase thermo-mechanical fatigue behavior of dissimilar welded joints of conventional materials (ferritic and martensitic material T24 and T92) to nickel-based alloys (A617B and HR6W) by means of membrane test panels. The temperature and external load are varied in phase during the test, while the internal pressure remains constant. At the temperature range between 650°C and 750°C, it focuses on the creep behavior under multiaxial stress loading of similar and dissimilar welded joints of high temperature resistant nickel-based alloys (A740H, A617B, and HR6W) by means of a thick-walled-component test. In this case, the temperature, the external axial load, and the internal pressure remain constant during testing. Numerical simulations are used for the estimation of the axial component load in order to induce a meaningful damage evolution without causing a total component failure. Metallographic investigations after testing will provide support for understanding the damage mechanism and the influence of the thermo-mechanical load and multiaxiality on the microstructure change and on the creep and TMF- strength.Keywords: creep, creep-fatigue, component behaviour, weld joints, high temperature material behaviour, nickel-alloys, high temperature resistant steels
Procedia PDF Downloads 113241 Noncovalent Antibody-Nanomaterial Conjugates: A Simple Approach to Produce Targeted Nanomedicines
Authors: Nicholas Fletcher, Zachary Houston, Yongmei Zhao, Christopher Howard, Kristofer Thurecht
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One promising approach to enhance nanomedicine therapeutic efficacy is to include a targeting agent, such as an antibody, to increase accumulation at the tumor site. However, the application of such targeted nanomedicines remains limited, in part due to difficulties involved with biomolecule conjugation to synthetic nanomaterials. One approach recently developed to overcome this has been to engineer bispecific antibodies (BsAbs) with dual specificity, whereby one portion binds to methoxy polyethyleneglycol (mPEG) epitopes present on synthetic nanomedicines, while the other binds to molecular disease markers of interest. In this way, noncovalent complexes of nanomedicine core, comprising a hyperbranched polymer (HBP) of primarily mPEG, decorated with targeting ligands are able to be produced by simple mixing. Further work in this area has now demonstrated such complexes targeting the breast cancer marker epidermal growth factor receptor (EGFR) to show enhanced binding to tumor cells both in vitro and in vivo. Indeed the enhanced accumulation at the tumor site resulted in improved therapeutic outcomes compared to untargeted nanomedicines and free chemotherapeutics. The current work on these BsAb-HBP conjugates focuses on further probing antibody-nanomaterial interactions and demonstrating broad applicability to a range of cancer types. Herein are reported BsAb-HBP materials targeted towards prostate-specific membrane antigen (PSMA) and study of their behavior in vivo using ⁸⁹Zr positron emission tomography (PET) in a dual-tumor prostate cancer xenograft model. In this model mice bearing both PSMA+ and PSMA- tumors allow for PET imaging to discriminate between nonspecific and targeted uptake in tumors, and better quantify the increased accumulation following BsAb conjugation. Also examined is the potential for formation of these targeted complexes in situ following injection of individual components? The aim of this approach being to avoid undesirable clearance of proteinaceous complexes upon injection limiting available therapeutic. Ultimately these results demonstrate BsAb functionalized nanomaterials as a powerful and versatile approach for producing targeted nanomedicines for a variety of cancers.Keywords: bioengineering, cancer, nanomedicine, polymer chemistry
Procedia PDF Downloads 135240 Morphological and Molecular Evaluation of Dengue Virus Serotype 3 Infection in BALB/c Mice Lungs
Authors: Gabriela C. Caldas, Fernanda C. Jacome, Arthur da C. Rasinhas, Ortrud M. Barth, Flavia B. dos Santos, Priscila C. G. Nunes, Yuli R. M. de Souza, Pedro Paulo de A. Manso, Marcelo P. Machado, Debora F. Barreto-Vieira
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The establishment of animal models for studies of DENV infections has been challenging, since circulating epidemic viruses do not naturally infect nonhuman species. Such studies are of great relevance to the various areas of dengue research, including immunopathogenesis, drug development and vaccines. In this scenario, the main objective of this study is to verify possible morphological changes, as well as the presence of antigens and viral RNA in lung samples from BALB/c mice experimentally infected with an epidemic and non-neuroadapted DENV-3 strain. Male BALB/c mice, 2 months old, were inoculated with DENV-3 by intravenous route. After 72 hours of infection, the animals were euthanized and the lungs were collected. Part of the samples was processed by standard technique for analysis by light and transmission electronic microscopies and another part was processed for real-time PCR analysis. Morphological analyzes of lungs from uninfected mice showed preserved tissue areas. In mice infected with DENV-3, the analyzes revealed interalveolar septum thickening with presence of inflammatory infiltrate, foci of alveolar atelectasis and hyperventilation, bleeding foci in the interalveolar septum and bronchioles, peripheral capillary congestion, accumulation of fluid in the blood capillary, signs of interstitial cell necrosis presence of platelets and mononuclear inflammatory cells circulating in the capillaries and/or adhered to the endothelium. In addition, activation of endothelial cells, platelets, mononuclear inflammatory cell and neutrophil-type polymorphonuclear inflammatory cell evidenced by the emission of cytoplasmic membrane prolongation was observed. DEN-like particles were seen in the cytoplasm of endothelial cells. The viral genome was recovered from 3 in 12 lung samples. These results demonstrate that the BALB / c mouse represents a suitable model for the study of the histopathological changes induced by DENV infection in the lung, with tissue alterations similar to those observed in human cases of DEN.Keywords: BALB/c mice, dengue, histopathology, lung, ultrastructure
Procedia PDF Downloads 250239 Study of the Combinatorial Impact of Substrate Properties on Mesenchymal Stem Cell Migration Using Microfluidics
Authors: Nishanth Venugopal Menon, Chuah Yon Jin, Samantha Phey, Wu Yingnan, Zhang Ying, Vincent Chan, Kang Yuejun
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Cell Migration is a vital phenomenon that the cells undergo in various physiological processes like wound healing, disease progression, embryogenesis, etc. Cell migration depends primarily on the chemical and physical cues available in the cellular environment. The chemical cue involves the chemokines secreted and gradients generated in the environment while physical cues indicate the impact of matrix properties like nanotopography and stiffness on the cells. Mesenchymal Stem Cells (MSCs) have been shown to have a role wound healing in vivo and its migration to the site of the wound has been shown to have a therapeutic effect. In the field of stem cell based tissue regeneration of bones and cartilage, one approach has been to introduce scaffold laden with MSCs into the site of injury to enable tissue regeneration. In this work, we have studied the combinatorial impact of the substrate physical properties on MSC migration. A microfluidic in vitro model was created to perform the migration studies. The microfluidic model used is a three compartment device consisting of two cell seeding compartments and one migration compartment. Four different PDMS substrates with varying substrate roughness, stiffness and hydrophobicity were created. Its surface roughness and stiffness was measured using Atomic Force Microscopy (AFM) while its hydrphobicity was measured from the water contact angle using an optical tensiometer. These PDMS substrates are sealed to the microfluidic chip following which the MSCs are seeded and the cell migration is studied over the period of a week. Cell migration was quantified using fluorescence imaging of the cytoskeleton (F-actin) to find out the area covered by the cells inside the migration compartment. The impact of adhesion proteins on cell migration was also quantified using a real-time polymerase chain reaction (qRT PCR). These results suggested that the optimal substrate for cell migration would be one with an intermediate level of roughness, stiffness and hydrophobicity. A higher or lower value of these properties affected cell migration negatively. These observations have helped us in understanding that different substrate properties need to be considered in tandem, especially while designing scaffolds for tissue regeneration as cell migration is normally impacted by the combinatorial impact of the matrix. These observations may lead us to scaffold optimization in future tissue regeneration applications.Keywords: cell migration, microfluidics, in vitro model, stem cell migration, scaffold, substrate properties
Procedia PDF Downloads 552238 Assessment of the Effect of Ethanolic Leaf Extract of Annona squamosa L. on Den Induced Hepatocellular Carcinoma in Experimental Animals
Authors: Vanitha Varadharaj, Vijalakshmi Krishnamurthy
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Annona squamosa Linn, commonly known as Sugar apple, belonging to the family Annonaceae, is said to show varied medicinal effects, including insecticide, antiovulatory and abortifacient. The alkaloid and flavonoids present in Annona squamosa leaf has proved to have antioxidant activity. The present work has been planned to investigate the effect of ethanolic leaf extract of Annona squamosa leaf on Den Induced wistar albino rats. The study was carried out to analyze the biochemical Parmeters like Total Proteins, Bilirubin, Enzymatic and Non –Enzymatic enzymes, Marker enzymes and Tumor markers in serum and also the histopathological studies in liver is carried out in control and DEN induced rats. Supplementation of ELAS (Ethanolic Leaf Extract Of Annona squamosa) reduced the liver weight and also reduced the tumour incidence. Chemoprevention group showed near normal values of bilirubin when compared with the control rats. Total protein was decreased in the cancer bearing group and on treatment with the extract the levels of protein were restored. Both in pre and post treatment group, the activities of enzymatic antioxidants such as superoxide dismutase, catalase, and Glutathione peroxidase were increased but in pre treated animals it was more effective than post treated animals. The non- enzymatic antioxidants such as vitamin C and vitamin E were brought back to normal level significantly in post and pre treated animals. Activities of marker enzymes such as SGOT, SGPT, ALP, γ GT were significantly elevated in the serum of cancer animals and the values returned to normal after treatment with the extract suggesting the hepato protective effect of the extract. Lipid peroxide was found to be elevated in the cancer induced group. This condition was brought back to the normal in the pre and post treated animals with ELAS. Histological examination also confirmed the anti- carcinogenic potential of ELAS, Cancer induced groups had a triple fold increase in their AFP values when compared to other groups. DEN treatment increased the level of AFP expression while ELAS partially counteracted the effect of it. So the scientific validation obtained from this study may pave way to many budding scientists to find new drugs from Annona squamosa for various ailments.Keywords: annona squamosa, biochemical parmeters, cancer, leaf extract
Procedia PDF Downloads 328237 Flexible Coupling between Gearbox and Pump (High Speed Machine)
Authors: Naif Mohsen Alharbi
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This paper present failure occurred on flexible coupling installed at oil anf gas operation. Also it presents maintenance ideas implemented on the flexible coupling installed to transmit high torque from gearbox to pump. Basically, the machine train is including steam turbine which drives the pump and there is gearbox located in between for speed reduction. investigation are identifying the root causes, solving and developing the technology designs or bad actor. This report provides the study intentionally for continues operation optimization, utilize the advanced opportunity and implement a improvement. Objective: The main objectives of the investigation are identifying the root causes, solving and developing the technology designs or bad actor. Ultimately, fulfilling the operation productivity, also ensuring better technology, quality and design by solutions. This report provides the study intentionally for continues operation optimization, utilize the advanced opportunity and implemet improvement. Method: The method used in this project was a very focused root cause analysis procedure that incorporated engineering analysis and measurements. The analysis method extensively covers the measuring of the complete coupling dimensions. Including the membranes thickness, hubs, bore diameter and total length, dismantle flexible coupling to diagnose how deep the coupling has been affected. Also, defining failure modes, so that the causes could be identified and verified. Moreover, Vibration analysis and metallurgy test. Lastly applying several solutions by advanced tools (will be mentioned in detail). Results and observation: Design capacity: Coupling capacity is an inadequate to fulfil 100% of operating conditions. Therefore, design modification of service factor to be at least 2.07 is crucial to address this issue and prevent recurrence of similar scenario, especially for the new upgrading project. Discharge fluctuation: High torque flexible coupling encountered during the operation. Therefore, discharge valve behaviour, tuning, set point and general conditions revaluated and modified subsequently, it can be used as baseline for upcoming Coupling design project. Metallurgy test: Material of flexible coupling membrane (discs) tested at the lab, for a detailed metallurgical investigation, better material grade has been selected for our operating conditions,Keywords: high speed machine, reliabilty, flexible coupling, rotating equipment
Procedia PDF Downloads 59236 Functional Performance of Unpaved Roads Reinforced with Treated Coir Geotextiles
Authors: Priya Jaswal, Vivek, S. K. Sinha
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One of the most important and complicated factors influencing the functional performance of unpaved roads is traffic loading. The complexity of traffic loading is caused by the variable magnitude and frequency of load, which causes unpaved roads to fail prematurely. Unpaved roads are low-volume roads, and as peri-urbanization increases, unpaved roads act as a means to boost the rural economy. This has also increased traffic on unpaved roads, intensifying the issue of settlement, rutting, and fatigue failure. This is a major concern for unpaved roads built on poor subgrade soil, as excessive rutting caused by heavy loads can cause driver discomfort, vehicle damage, and an increase in maintenance costs. Some researchers discovered that when a consistent static load is exerted as opposed to a rapidly changing load, the rate of deformation of unpaved roads increases. Previously, some of the most common methods for overcoming the problem of rutting and fatigue failure included chemical stabilisation, fibre reinforcement, and so on. However, due to their high cost, engineers' attention has shifted to geotextiles which are used as reinforcement in unpaved roads. Geotextiles perform the function of filtration, lateral confinement of base material, vertical restraint of subgrade soil, and the tension membrane effect. The use of geotextiles in unpaved roads increases the strength of unpaved roads and is an economically viable method because it reduces the required aggregate thickness, which would need less earthwork, and is thus recommended for unpaved road applications. The majority of geotextiles used previously were polymeric, but with a growing awareness of sustainable development to preserve the environment, researchers' focus has shifted to natural fibres. Coir is one such natural fibre that possesses the advantage of having a higher tensile strength than other bast fibres, being eco-friendly, low in cost, and biodegradable. However, various researchers have discovered that the surface of coir fibre is covered with various impurities, voids, and cracks, which act as a plane of weakness and limit the potential application of coir geotextiles. To overcome this limitation, chemical surface modification of coir geotextiles is widely accepted by researchers because it improves the mechanical properties of coir geotextiles. The current paper reviews the effect of using treated coir geotextiles as reinforcement on the load-deformation behaviour of a two-layered unpaved road model.Keywords: coir, geotextile, treated, unpaved
Procedia PDF Downloads 89235 A Review on Silicon Based Induced Resistance in Plants against Insect Pests
Authors: Asim Abbasi, Muhammad Sufyan, Muhammad Kamran, Iqra
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Development of resistance in insect pests against various groups of insecticides has prompted the use of alternative integrated pest management approaches. Among these induced host plant resistance represents an important strategy as it offers a practical, cheap and long lasting solution to keep pests populations below economic threshold level (ETL). Silicon (Si) has a major role in regulating plant eco-relationship by providing strength to the plant in the form of anti-stress mechanism which was utilized in coping with the environmental extremes to get a better yield and quality end produce. Among biotic stresses, insect herbivore signifies one class against which Si provide defense. Silicon in its neutral form (H₄SiO₄) is absorbed by the plants via roots through an active process accompanied by the help of different transporters which were located in the plasma membrane of root cells or by a passive process mostly regulated by transpiration stream, which occurs via the xylem cells along with the water. Plants tissues mainly the epidermal cell walls are the sinks of absorbed silicon where it polymerizes in the form of amorphous silica or monosilicic acid. The noteworthy function of this absorbed silicon is to provide structural rigidity to the tissues and strength to the cell walls. Silicon has both direct and indirect effects on insect herbivores. Increased abrasiveness and hardness of epidermal plant tissues and reduced digestibility as a result of deposition of Si primarily as phytoliths within cuticle layer is now the most authenticated mechanisms of Si in enhancing plant resistance to insect herbivores. Moreover, increased Si content in the diet also impedes the efficiency by which insects transformed consumed food into the body mass. The palatability of food material has also been changed by Si application, and it also deters herbivore feeding for food. The production of defensive compounds of plants like silica and phenols have also been amplified by the exogenous application of silicon sources which results in reduction of the probing time of certain insects. Some studies also highlighted the role of silicon at the third trophic level as it also attracts natural enemies of insects attacking the crop. Hence, the inclusion of Si in pest management approaches can be a healthy and eco-friendly tool in future.Keywords: defensive, phytoliths, resistance, stresses
Procedia PDF Downloads 183234 Angiomotin Regulates Integrin Beta 1-Mediated Endothelial Cell Migration and Angiogenesis
Authors: Yuanyuan Zhang, Yujuan Zheng, Giuseppina Barutello, Sumako Kameishi, Kungchun Chiu, Katharina Hennig, Martial Balland, Federica Cavallo, Lars Holmgren
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Angiogenesis describes that new blood vessels migrate from pre-existing ones to form 3D lumenized structure and remodeling. During directional migration toward the gradient of pro-angiogenic factors, the endothelial cells, especially the tip cells need filopodia to sense the environment and exert the pulling force. Of particular interest are the integrin proteins, which play an essential role in focal adhesion in the connection between migrating cells and extracellular matrix (ECM). Understanding how these biomechanical complexes orchestrate intrinsic and extrinsic forces is important for our understanding of the underlying mechanisms driving angiogenesis. We have previously identified Angiomotin (Amot), a member of Amot scaffold protein family, as a promoter for endothelial cell migration in vitro and zebrafish models. Hence, we established inducible endothelial-specific Amot knock-out mice to study normal retinal angiogenesis as well as tumor angiogenesis. We found that the migration ratio of the blood vessel network to the edge was significantly decreased in Amotec- retinas at postnatal day 6 (P6). While almost all the Amot defect tip cells lost migration advantages at P7. In consistence with the dramatic morphology defect of tip cells, there was a non-autonomous defect in astrocytes, as well as the disorganized fibronectin expression pattern correspondingly in migration front. Furthermore, the growth of transplanted LLC tumor was inhibited in Amot knockout mice due to fewer vasculature involved. By using MMTV-PyMT transgenic mouse model, there was a significantly longer period before tumors arised when Amot was specifically knocked out in blood vessels. In vitro evidence showed that Amot binded to beta-actin, Integrin beta 1 (ITGB1), Fibronectin, FAK, Vinculin, major focal adhesion molecules, and ITGB1 and stress fibers were distinctly induced by Amot transfection. Via traction force microscopy, the total energy (force indicater) was found significantly decreased in Amot knockdown cells. Taken together, we propose that Amot is a novel partner of the ITGB1/Fibronectin protein complex at focal adhesion and required for exerting force transition between endothelial cell and extracellular matrix.Keywords: angiogenesis, angiomotin, endothelial cell migration, focal adhesion, integrin beta 1
Procedia PDF Downloads 228233 Oat βeta Glucan Attenuates the Development of Atherosclerosis and Improves the Intestinal Barrier Function by Reducing Bacterial Endotoxin Translocation in APOE-/- MICE
Authors: Dalal Alghawas, Jetty Lee, Kaisa Poutanen, Hani El-Nezami
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Oat β-glucan a water soluble non starch linear polysaccharide has been approved as a cholesterol lowering agent by various food safety administrations and is commonly used to reduce the risk of heart disease. The molecular weight of oat β-glucan can vary depending on the extraction and fractionation methods. It is not clear whether the molecular weight has a significant impact at reducing the acceleration of atherosclerosis. The aim of this study was to investigate three different oat β-glucan fractionations on the development of atherosclerosis in vivo. With special focus on plaque stability and the intestinal barrier function. To test this, ApoE-/- female mice were fed a high fat diet supplemented with oat bran, high molecular weight (HMW) oat β-glucan fractionate and low molecular weight (LMW) oat β-glucan fractionate for 16 weeks. Atherosclerosis risk markers were measured in the plasma, heart and aortic tree. Plaque size was measured in the aortic root and aortic tree. ICAM-1, VCAM-1, E-Selectin, P-Selectin, protein levels were assessed from the aortic tree to determine plaque stability at 16 weeks. The expression of p22phox at the aortic root was evaluated to study the NADPH oxidase complex involved in nitric oxide bioavailability and vascular elasticity. The tight junction proteins E-cadherin and beta-catenin from western blot analyses were analysed as an intestinal barrier function test. Plasma LPS, intestinal D-lactate levels and hepatic FMO gene expression were carried out to confirm whether the compromised intestinal barrier lead to endotoxemia. The oat bran and HMW oat β-glucan diet groups were more effective than the LMW β-glucan diet group at reducing the plaque size and showed marked improvements in plaque stability. The intestinal barrier was compromised for all the experimental groups however the endotoxemia levels were higher in the LMW β-glucan diet group. The oat bran and HMW oat β-glucan diet groups were more effective at attenuating the development of atherosclerosis. Reasons for this could be due to the LMW oat β-glucan diet group’s low viscosity in the gut and the inability to block the reabsorption of cholesterol. Furthermore the low viscosity may allow more bacterial endotoxin translocation through the impaired intestinal barrier. In future food technologists should carefully consider how to incorporate LMW oat β-glucan as a health promoting food.Keywords: Atherosclerosis, beta glucan, endotoxemia, intestinal barrier function
Procedia PDF Downloads 412232 Refractory Cardiac Arrest: Do We Go beyond, Do We Increase the Organ Donation Pool or Both?
Authors: Ortega Ivan, De La Plaza Edurne
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Background: Spain and other European countries have implemented Uncontrolled Donation after Cardiac Death (uDCD) programs. After 15 years of experience in Spain, many things have changed. Recent evidence and technical breakthroughs achieved in resuscitation are relevant for uDCD programs and raise some ethical concerns related to these protocols. Aim: To rethink current uDCD programs in the light of recent evidence on available therapeutic procedures applicable to victims of out-of-hospital cardiac arrest (OHCA). To address the following question: What is the current standard of treatment owed to victims of OHCA before including them in an uDCD protocol? Materials and Methods: Review of the scientific and ethical literature related to both uDCD programs and innovative resuscitation techniques. Results: 1) The standard of treatment received and the chances of survival of victims of OHCA depend on whether they are classified as Non-Heart Beating Patients (NHBP) or Non-Heart-Beating-Donors (NHBD). 2) Recent studies suggest that NHBPs are likely to survive, with good quality of life, if one or more of the following interventions are performed while ongoing CPR -guided by suspected or known cause of OHCA- is maintained: a) direct access to a Cath Lab-H24 or/and to extra-corporeal life support (ECLS); b) transfer in induced hypothermia from the Emergency Medical Service (EMS) to the ICU; c) thrombolysis treatment; d) mobile extra-corporeal membrane oxygenation (mini ECMO) instituted as a bridge to ICU ECLS devices. 3) Victims of OHCA who cannot benefit from any of these therapies should be considered as NHBDs. Conclusion: Current uDCD protocols do not take into account recent improvements in resuscitation and need to be adapted. Operational criteria to distinguish NHBDs from NHBP should seek a balance between the technical imperative (to do whatever is possible), considerations about expected survival with quality of life, and distributive justice (costs/benefits). Uncontrolled DCD protocols can be performed in a way that does not hamper the legitimate interests of patients, potential organ donors, their families, the organ recipients, and the health professionals involved in these processes. Families of NHBDs’ should receive information which conforms to the ethical principles of respect of autonomy and transparency.Keywords: uncontrolled donation after cardiac death resuscitation, refractory cardiac arrest, out of hospital cardiac, arrest ethics
Procedia PDF Downloads 231231 Behavior of GRS Abutment Facing under Variable Cycles of Lateral Excitation through Physical Model Tests
Authors: Ashutosh Verma, Satyendra Mittal
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Numerous geosynthetic reinforced soil (GRS) abutment failures over the years have been attributed to the loss of strength at the facing-reinforcement interface due to seasonal thermal expansion/contraction of the bridge deck. This causes excessive settlement below the bridge seat, causing bridge bumps along the approach road which reduces the design life of any abutment. Before designers while choosing the type of facing, a broad range of facing configurations are undoubtedly available. Generally speaking, these configurations can be divided into three groups: modular (panels/block), continuous, and full height rigid (FHR). The purpose of the current study is to use 1g physical model tests under serviceable cyclic lateral displacements to experimentally investigate the behaviour of these three facing classifications. To simulate field behaviour, a field instrumented GRS abutment prototype was modeled into a N scaled down 1g physical model (N = 5) with adjustable facing arrangements to represent these three facing classifications. For cyclic lateral displacement (d/H) of top facing at loading rate of 1mm/min, the peak earth pressure coefficient (K) on the facing and vertical settlement of the footing (s/B) at 25, 50, 75 and 100 cycles have been measured. For a constant footing offset of x/H = 0.1, three forms of cyclic displacements have been performed to simulate active condition (CA), passive condition (CP), and active-passive condition (CAP). The findings showed that when reinforcements are integrated into the wall along with presence of gravel gabions i.e. FHR design, a rather substantial earth pressure occurs over the facing. Despite this, the FHR facing's continuous nature works in conjunction with the reinforcements' membrane resilience to reduce footing settlement. On the other hand, the pressure over the wall is released upon lateral excitation by the relative displacement between the panels in modular facing reducing the connection strength at the interface and leading to greater settlements below footing. On the contrary, continuous facing do not exhibit relative displacement along the depth of facing rather fails through rotation about the base, which extends the zone of active failure in the backfill leading to large depressions in the backfill region around the bridge seat. Conservatively, FHR facing shows relatively stable responses under lateral cyclic excitations as compared to modular or continuous type of abutment facing.Keywords: GRS abutments, 1g physical model, full height rigid, cyclic lateral displacement
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