Search results for: pharmaceutical pollutants degradation

Authors: Maria C. Proença, Maria T. Rebelo, Marília Antunes, Maria J. Alves, Hugo Osório, Sofia Cunha, REVIVE team

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The environment of the mosquitoes complex Culex pipiens sl. in Portugal mainland is evaluated based in its abundance, using a data set georeferenced, collected during seven years (2006-2012) from May to October. The suitability of the different regions can be delineated using the relative abundance areas; the suitablility index is directly proportional to disease transmission risk and allows focusing mitigation measures in order to avoid outbreaks of vector-borne diseases. The interest in the Culex pipiens complex is justified by its medical importance: the females bite all warm-blooded vertebrates and are involved in the circulation of several arbovirus of concern to human health, like West Nile virus, iridoviruses, rheoviruses and parvoviruses. The abundance of Culex pipiens mosquitoes were documented systematically all over the territory by the local health services, in a long duration program running since 2006. The environmental factors used to characterize the vector habitat are land use/land cover, distance to cartographed water bodies, altitude and latitude. Focus will be on the mosquito females, which gonotrophic cycle mate-bloodmeal-oviposition is responsible for the virus transmission; its abundance is the key for the planning of non-aggressive prophylactic countermeasures that may eradicate the transmission risk and simultaneously avoid chemical ambient degradation. Meteorological parameters such as: air relative humidity, air temperature (minima, maxima and mean daily temperatures) and daily total rainfall were gathered from the weather stations network for the same dates and crossed with the standardized females’ abundance in a geographic information system (GIS). Mean capture and percentage of above average captures related to each variable are used as criteria to compute a threshold for each meteorological parameter; the difference of the mean capture above/below the threshold was statistically assessed. The meteorological parameters measured at the net of weather stations all over the country are averaged by month and interpolated to produce raster maps that can be segmented according to the meaningful thresholds for each parameter. The intersection of the maps of all the parameters obtained for each month show the evolution of the suitable meteorological conditions through the mosquito season, considered as May to October, although the first and last month are less relevant. In parallel, mean and above average captures were related to the physiographic parameters – the land use/land cover classes most relevant in each month, the altitudes preferred and the most frequent distance to water bodies, a factor closely related with the mosquito biology. The maps produced with these results were crossed with the meteorological maps previously segmented, in order to get an index of suitability for the complex Culex pipiens evaluated all over the country, and its evolution from the beginning to the end of the mosquitoes season.

Keywords: suitability index, Culex pipiens, habitat evolution, GIS model

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Authors: Beihai Xiong, Dengke Hua, Wouter Hendriks, Wilbert Pellikaan

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To improve the energy status of dairy cows in the early lactation, lots of jobs have been done on adjusting the starch to fiber ratio in the diet. As a complex ecosystem, the rumen contains a large population of microorganisms which plays a crucial role in feed degradation. Further study on the microbiota alterations and metabolic changes under different dietary energy sources is essential and valuable to better understand the function of the ruminal microorganisms and thereby to optimize the rumen function and enlarge feed efficiency. The present study will focus on the effects of two glucogenic diets (G: ground corn and corn silage; S: steam-flaked corn and corn silage) and a lipogenic diet (L: sugar beet pulp and alfalfa silage) on rumen fermentation, gas production, the ruminal microbiota and metabolome, and also their correlations in vitro. The gas production was recorded consistently, and the gas volume and producing rate at times 6, 12, 24, 48 h were calculated separately. The fermentation end-products were measured after fermenting for 48 h. The ruminal bacteria and archaea communities were determined by 16S RNA sequencing technique, the metabolome profile was tested through LC-MS methods. Compared to the diet G and S, the L diet had a lower dry matter digestibility, propionate production, and ammonia-nitrogen concentration. The two glucogenic diets performed worse in controlling methane and lactic acid production compared to the L diet. The S diet produced the greatest cumulative gas volume at any time points during incubation compared to the G and L diet. The metabolic analysis revealed that the lipid digestion was up-regulated by the diet L than other diets. On the subclass level, most metabolites belonging to the fatty acids and conjugates were higher, but most metabolites belonging to the amino acid, peptides, and analogs were lower in diet L than others. Differences in rumen fermentation characteristics were associated with (or resulting from) changes in the relative abundance of bacterial and archaeal genera. Most highly abundant bacteria were stable or slightly influenced by diets, while several amylolytic and cellulolytic bacteria were sensitive to the dietary changes. The L diet had a significantly higher number of cellulolytic bacteria, including the genera of Ruminococcus, Butyrivibrio, Eubacterium, Lachnospira, unclassified Lachnospiraceae, and unclassified Ruminococcaceae. The relative abundances of amylolytic bacteria genera including Selenomonas_1, Ruminobacter, and Succinivibrionaceae_UCG-002 were higher in diet G and S. These affected bacteria was also proved to have high associations with certain metabolites. The Selenomonas_1 and Succinivibrionaceae_UCG-002 may contribute to the higher propionate production in the diet G and S through enhancing the succinate pathway. The results indicated that the two glucogenic diets had a greater extent of gas production, a higher dry matter digestibility, and produced more propionate than diet L. The steam-flaked corn did not show a better performance on fermentation end-products than ground corn. This study has offered a deeper understanding of ruminal microbial functions which could assistant the improvement in rumen functions and thereby in the ruminant production.

Keywords: gas production, metabolome, microbiota, rumen fermentation

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Authors: Indra Bahadur Chand

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This paper will focus on the need for development and application of global protocols and policy in planning, designing, development, and management of systems of eco-towns and eco-villages so that sustainable development will be assured from the perspective of environmental, economical, peace, and harmonized social dynamics. This perspective is essential for the development of civilized and eco-friendly human settlements in the town and rural areas of the nation that will be a milestone for developing a happy and sustainable lifestyle of rural and urban communities of the nation. The urban population of most of the town of developing economies has been tremendously increasing, whereas rural people have been tremendously migrating for the past three decades. Consequently, the urban lifestyle in most towns has stressed in terms of environmental pollution, water crisis, congested traffic, energy crisis, food crisis, and unemployment. Eco-towns and villages should be developed where lifestyle of all residents is sustainable and happy. Built up environment of settlement should reduce and minimize the problems of non ecological CO2 emissions, unbalanced utilization of natural resources, environmental degradation, natural calamities, ecological imbalance, energy crisis, water scarcity, waste management, food crisis, unemployment, deterioration of cultural heritage, social, the ratio among the public and private land ownership, ratio of land covered with vegetation and area of settlement, the ratio of people in the vehicles and foot, the ratio of people employed outside of town and village, ratio of resources recycling of waste materials, water consumption level, the ratio of people and vehicles, ratio of the length of the road network and area of town/villages, a ratio of renewable energy consumption with total energy, a ratio of religious/recreational area out of the total built-up area, the ratio of annual suicide case out of total people, a ratio of annual injured and death out of total people from a traffic accident, a ratio of production of agro foods within town out of total food consumption will be used to assist in designing and monitoring of each eco-towns and villages. An eco-town and villages should be planned and developed to offer sustainable infrastructure and utilities that maintain CO2 level in individual homes and settlements, home energy use, transport, food and consumer goods, water supply, waste management, conservation of historical heritages, healthy neighborhood, conservation of natural landscape, conserving bio-diversity and developing green infrastructures. Eco-towns and villages should be developed on the basis of master planning and architecture that affect and define the settlement and its form. Master planning and engineering should focus in delivering the sustainability criteria of eco towns and eco village. This will involve working with specific landscape and natural resources of locality.

Keywords: eco-town, ecological habitation, master plan, sustainable development

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Authors: Tauseef Ahmad, Muhammad Ishaq, Mathew Eapen, Ahyoung Park, Sam Karpiniec, Vanni Caruso, Rajaraman Eri

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Fucoidans are complex, fucose-rich sulfated polymers discovered in brown seaweeds. Fucoidans are popular around the world, particularly in the nutraceutical and pharmaceutical industries, due to their promising medicinal properties. Fucoidans have been shown to have a variety of biological activities, including anti-inflammatory effects. They are known to inhibit inflammatory processes through a variety of mechanisms, including enzyme inhibition and selectin blockade. Inflammation is a part of the complicated biological response of living systems to damaging stimuli, and it plays a role in the pathogenesis of a variety of disorders, including arthritis, inflammatory bowel disease, cancer, and allergies. In the current investigation, various fucoidan extracts from Undaria pinnatifida, Fucus vesiculosus, Macrocystis pyrifera, Ascophyllum nodosum, and Laminaria japonica were assessed for inhibition of pro-inflammatory cytokine production (TNF-α, IL-1β, and IL-6) in LPS induced human macrophage cell line (THP-1) and human peripheral blood mononuclear cells (PBMCs). Furthermore, we also sought to catalogue these extracts based on their anti-inflammatory effects in the current in-vitro cell model. Materials and Methods: To assess the cytotoxicity of fucoidan extracts, MTT (3-[4,5-dimethylthiazol-2-yl]-2,5, -diphenyltetrazolium bromide) cell viability assay was performed. Furthermore, a dose-response for fucoidan extracts was performed in LPS induced THP-1 cells and PBMCs after pre-treatment for 24 hours, and levels of TNF-α, IL-1β, and IL-6 cytokines were measured using Enzyme-Linked Immunosorbent Assay (ELISA). Results: The MTT cell viability assay demonstrated that fucoidan extracts exhibited no evidence of cytotoxicity in THP-1 cells or PBMCs after 48 hours of incubation. The results of the sandwich ELISA revealed that all fucoidan extracts suppressed cytokine production in LPS-stimulated PBMCs and human THP-1 cells in a dose-dependent manner. Notably, at lower concentrations, the lower molecular fucoidan (5-30 kDa) extract from Macrocystis pyrifera was a highly efficient inhibitor of pro-inflammatory cytokines. Fucoidan extracts from all species including Undaria pinnatifida, Fucus vesiculosus, Macrocystis pyrifera, Ascophyllum nodosum, and Laminaria japonica exhibited significant anti-inflammatory effects. These findings on several fucoidan extracts provide insight into strategies for improving their efficacy against inflammation-related diseases. Conclusion: In the current research, we have successfully catalogued several fucoidan extracts based on their efficiency in LPS-induced macrophages and PBMCs in downregulating the key pro-inflammatory cytokines (TNF-, IL-1 and IL-6), which are prospective targets in human inflammatory illnesses. Further research would provide more information on the mechanism of action, allowing it to be tested for therapeutic purposes as an anti-inflammatory medication.

Keywords: fucoidan, PBMCs, THP-1, TNF-α, IL-1β, IL-6, inflammation

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Authors: Prescilla Lambert, Christophe Roos, Mounim Lebrini

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For several years, the Caribbean islands and West Africa have had to deal with the massive arrival of the brown seaweed Sargassum. Overall, this macroalgae, which constitutes a habitat for a great diversity of marine organisms, is also an additional stress factor for the marine environment (e.g., coral reefs). In addition, the accumulation followed by the significant decomposition of the Sargassum spp. biomass on the coast leads to the release of toxic gases (H₂S and NH₃), which calls into question the functioning of the economic, health and tourist life of the island and the other interested territories. Originally, these algae are formed by the eutrophication of the oceans accentuated by global warming. Unfortunately, scientists predict a significant recurrence of these Sargassum strandings for years to come. It is therefore more than necessary to find solutions by putting in place a sustainable management plan for this phenomenon. Martinique, a small island in the Caribbean arc, is one of the many areas impacted by Sargassum seaweed strandings. Since 2011, there has been a constant increase in the degradation of the materials present in this region, largely due to toxic/corrosive gases released by the algae decomposition. In order to protect the structures and the vulnerable building materials while limiting the use of synthetic/petroleum based molecules as much as possible, research is being conducted on molecules of natural origin. Thus, thanks to the chemical composition, which comprise molecules with interesting properties, algae such as Sargassum could potentially help to solve many issues. Therefore, this study focuses on the green extraction and characterization of molecules from the species Sargassum fluitans and Sargassum natans present in Martinique. The secondary metabolites found in these extracts showed variability in yield rates due to local climatic conditions. The tests carried out shed light on the anticorrosive and antibacterial potential of the algae. These extracts can thus be described as natural inhibitors. The effect of variation in inhibitor concentrations was tested in electrochemistry using electrochemical impedance spectroscopy and polarization curves. The analysis of electrochemical results obtained by direct immersion in the extracts and self-assembled molecular layers (SAMs) for Sargassum fluitans III, Sargassum natans I and VIII species was conclusive in acid and alkaline environments. The excellent results obtained reveal an inhibitory efficacy of 88% at 50mg/L for the crude extract of Sargassum fluitans III and efficacies greater than 97% for the chemical families of Sargassum fluitans III. Similarly, microbiological tests also suggest a bactericidal character. Results for Sargassum fluitans III crude extract show a minimum inhibitory concentration (MIC) of 0.005 mg/mL on Gram-negative bacteria and a MIC greater than 0.6 mg/mL on Gram-positive bacteria. These results make it possible to consider the management of local and international issues while valuing a biomass rich in biodegradable molecules. The next step in this study will therefore be the evaluation of the toxicity of Sargassum spp..

Keywords: Sargassum, secondary metabolites, anticorrosive, antibacterial, natural inhibitors

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Authors: Godfrey Omulo, Regina Birner, Karlheinz Koller, Thomas Daum

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Conservation agriculture (CA) as a climate-resilient and sustainable practice have been carried out for over three decades in Zambia. However, its continued promotion and adoption has been predominantly on a small-scale basis. Despite the plethora of scholarship pointing to the positive benefits of CA in regard to enhanced yield, profitability, carbon sequestration and minimal environmental degradation, these have not stimulated commensurate agricultural extensification desired for Zambia. The objective of this study was to investigate the potential differences between mechanized conventional and conservation tillage practices on operation time, fuel consumption, labor costs, soil moisture retention, soil temperature and crop yield. An on-farm mechanized conservation agriculture (MCA) experiment arranged in a randomized complete block design with four replications was used. The research was conducted on a 15 ha of sandy loam rainfed land: soybeans on 7ha with plot dimensions of 24 m by 210 m and maize on 8ha with plot dimensions of 24 m by 250 m. The three tillage treatments were: residue burning followed by disc harrowing, ripping tillage and no-till. The crops were rotated in two subsequent seasons. All operations were done using a 60hp 2-wheel tractor, a disc harrow, a two-tine ripper and a two-row planter. Soil measurements and the agro-economic factors were recorded for two farming seasons. The season results showed that the yield of maize and soybeans under no-till and ripping tillage practices were not significantly different from the conventional burning and discing. But, there was a significant difference in soil moisture content between no-till (25.31SFU±2.77) and disced (11.91SFU±0.59) plots at depths from 10-60 cm. Soil temperature in no-till plots (24.59°C±0.91) was significantly lower compared to the disced plots (26.20°C±1.75) at the depths 15 cm and 45 cm. For maize, there was a significant difference in operation time between disc-harrowed (3.68hr/ha±1.27) and no-till (1.85hr/ha±0.04) plots, and a significant difference in cost of labor between disc-harrowed (45.45$/ha±19.56) and no-till (21.76$/ha) plots. There was no significant difference in fuel consumption between ripping and disc-harrowing and direct seeding. For soybeans, there was a significant difference in operation time between no-tillage (1.96hr/ha±0.31) and ripping (3.34hr/ha±0.53) and disc harrowing (3.30hr/ha±0.16). Further, fuel consumption and labor on no-till plots were significantly different from both the ripped and disc-harrowed plots. The high seed emergence percentage on maize disc-harrowed plot (93.75%±5.87) was not significantly different from ripping and no-till plots. Again, the high seed emergence percentage for the soybean ripped plot (93.75%±13.03) had no significant difference with discing and ripping. The results show that it is economically sound and timesaving to practice MCA and get viable yields compared to conventional farming. This research fills the gap on the potential of MCA in the context of Zambia and its profitability in incentivizing policymakers to invest in appropriate and sustainable machinery and implements for extensive agricultural production.

Keywords: climate-smart agriculture, labor cost, mechanized conservation agriculture, soil moisture, Zambia

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Authors: Srijata Mitra, Mobina Parveen, Pranab Roy, Narayan Chandra Chattopadhyay

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Chlorinated hydrocarbon can be a major pollution problem in groundwater as well as soil. Many people interact with these chemicals on daily accidentally or by professionally in the laboratory. One of the most common sources for Chlorinated hydrocarbon contamination of soil and groundwater are industrial effluents. The wide use and discharge of Trichloroethylene (TCE), a volatile chlorohydrocarbon from chemical industry, led to major water pollution in rural areas. TCE is an mainly used as an industrial metal degreaser in industries. Biotransformation of TCE to the potent carcinogen vinyl chloride (VC) by consortia of anaerobic bacteria might have role for the above purpose. For these reasons, the aim of current study was to isolate and characterized the genes involved in TCE metabolism and also to investigate the in silico study of those genes. To our knowledge, only one aromatic dioxygenase system, the toluene dioxygenase in Pseudomonas putida F1 has been shown to be involved in TCE degradation. This is first instance where Bacillus cereus group being used in biodegradation of trichloroethylene. A novel bacterial strain 2479 was isolated from oil depot site at Rajbandh, Durgapur (West Bengal, India) by enrichment culture technique. It was identified based on polyphasic approach and ribotyping. The bacterium was gram positive, rod shaped, endospore forming and capable of degrading trichloroethylene as the sole carbon source. On the basis of phylogenetic data and Fatty Acid Methyl Ester Analysis, strain 2479 should be placed within the genus Bacillus and species cereus. However, the present isolate (strain 2479) is unique and sharply different from the usual Bacillus strains in its biodegrading nature. Fujiwara test was done to estimate that the strain 2479 could degrade TCE efficiently. The gene for TCE biodegradation was PCR amplified from genomic DNA of Bacillus cereus 2479 by using todC1 gene specific primers. The 600bp amplicon was cloned into expression vector pUC I8 in the E. coli host XL1-Blue and expressed under the control of lac promoter and nucleotide sequence was determined. The gene sequence was deposited at NCBI under the Accession no. GU183105. In Silico approach involved predicting the physico-chemical properties of deduced Tce1 protein by using ProtParam tool. The tce1 gene contained 342 bp long ORF encoding 114 amino acids with a predicted molecular weight 12.6 kDa and the theoretical pI value of the polypeptide was 5.17, molecular formula: C559H886N152O165S8, total number of atoms: 1770, aliphatic index: 101.93, instability index: 28.60, Grand Average of Hydropathicity (GRAVY): 0.152. Three differentially expressed proteins (97.1, 40 and 30 kDa) were directly involved in TCE biodegradation, found to react immunologically to the antibodies raised against TCE inducible proteins in Western blot analysis. The present study suggested that cloned gene product (TCE1) was capable of degrading TCE as verified chemically.

Keywords: cloning, Bacillus cereus, in silico analysis, TCE

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Authors: Marek Vondra, Petr Bobák

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Vacuum evaporation is a reliable and well-proven technology with a wide application range which is frequently used in food, chemical or pharmaceutical industries. Recently, numerous remarkable studies have been carried out to investigate utilization of this technology in the area of wastewater treatment. One of the most successful applications of vacuum evaporation principal is connected with seawater desalination. Since 1950’s, multi-stage flash distillation (MSF) has been the leading technology in this field and it is still irreplaceable in many respects, despite a rapid increase in cheaper reverse-osmosis-based installations in recent decades. MSF plants are conveniently operated in countries with a fluctuating seawater quality and at locations where a sufficient amount of waste heat is available. Nowadays, most of the MSF research is connected with alternative heat sources utilization and with hybridization, i.e. merging of different types of desalination technologies. Some of the studies are concerned with basic principles of the static flash phenomenon, but only few scientists have lately focused on the fundamentals of continuous multi-stage evaporation. Limited measurement possibilities at operating plants and insufficiently equipped experimental facilities may be the reasons. The aim of the presented study was to design, construct and test an up-to-date test rig with an advanced measurement system which will provide real time monitoring options of all the important operational parameters under various conditions. The whole system consists of a conventionally designed MSF unit with 8 evaporation chambers, versatile heating circuit for different kinds of feed water (e.g. seawater, waste water), sophisticated system for acquisition and real-time visualization of all the related quantities (temperature, pressure, flow rate, weight, conductivity, pH, water level, power input), access to a wide spectrum of operational media (salt, fresh and softened water, steam, natural gas, compressed air, electrical energy) and integrated transparent features which enable a direct visual control of selected physical mechanisms (water evaporation in chambers, water level right before brine and distillate pumps). Thanks to the adjustable process parameters, it is possible to operate the test unit at desired operational conditions. This allows researchers to carry out statistical design and analysis of experiments. Valuable results obtained in this manner could be further employed in simulations and process modeling. First experimental tests confirm correctness of the presented approach and promise interesting outputs in the future. The presented experimental apparatus enables flexible and efficient research of the whole MSF process.

Keywords: design of experiment, multi-stage flash distillation, test rig, vacuum evaporation

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Authors: K. S. Rao, Ankesh Kumar

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The search for alternative energy sources to petroleum has increased these days because of increase in need and depletion of petroleum reserves. Therefore the importance of oil shales as an economically viable substitute has increased many folds in last 20 years. The technologies like hydro-fracturing have opened the field of oil extraction from these unconventional rocks. Oil shale is a compact laminated rock of sedimentary origin containing organic matter known as kerogen which yields oil when distilled. Oil shales are formed from the contemporaneous deposition of fine grained mineral debris and organic degradation products derived from the breakdown of biota. Conditions required for the formation of oil shales include abundant organic productivity, early development of anaerobic conditions, and a lack of destructive organisms. These rocks are not gown through the high temperature and high pressure conditions in Mother Nature. The most common approach for oil extraction is drastically breaking the bond of the organics which involves retorting process. The two approaches for retorting are surface retorting and in-situ processing. The most environmental friendly approach for extraction is In-situ processing. The three steps involved in this process are fracturing, injection to achieve communication, and fluid migration at the underground location. Upon heating (retorting) oil shale at temperatures in the range of 300 to 400°C, the kerogen decomposes into oil, gas and residual carbon in a process referred to as pyrolysis. Therefore it is very important to understand the physico-mechenical behavior of such rocks, to improve the technology for in-situ extraction. It is clear from the past research and the physical observations that these rocks will behave as an anisotropic rock so it is very important to understand the mechanical behavior under high pressure at different orientation angles for the economical use of these resources. By knowing the engineering behavior under above conditions will allow us to simulate the deep ground retorting conditions numerically and experimentally. Many researchers have investigate the effect of organic content on the engineering behavior of oil shale but the coupled effect of organic and inorganic matrix is yet to be analyzed. The favourable characteristics of Assam coal for conversion to liquid fuels have been known for a long time. Studies have indicated that these coals and carbonaceous shale constitute the principal source rocks that have generated the hydrocarbons produced from the region. Rock cores of the representative samples are collected by performing on site drilling, as coring in laboratory is very difficult due to its highly anisotropic nature. Different tests are performed to understand the petrology of these samples, further the chemical analyses are also done to exactly quantify the organic content in these rocks. The mechanical properties of these rocks are investigated by considering different anisotropic angles. Now the results obtained from petrology and chemical analysis are correlated with the mechanical properties. These properties and correlations will further help in increasing the producibility of these rocks. It is well established that the organic content is negatively correlated to tensile strength, compressive strength and modulus of elasticity.

Keywords: oil shale, producibility, hydro-fracturing, kerogen, petrology, mechanical behavior

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Authors: Leo Nnamdi Ozurumba-Dwight

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Single-cell genomic analytical systems have proved to be a platform to isolate bulk cells into selected single cells for genomic, proteomic, and related metabolomic studies. This is enabling systematic investigations of the level of heterogeneity in a diverse and wide pool of cell populations. Single cell technologies, embracing techniques such as high parameter flow cytometry, single-cell sequencing, and high-resolution images are playing vital roles in these investigations on messenger ribonucleic acid (mRNA) molecules and related gene expressions in tracking the nature and course of disease conditions. This entails targeted molecular investigations on unit cells that help us understand cell behavoiur and expressions, which can be examined for their health implications on the health state of patients. One of the vital good sides of single-cell RNA sequencing (scRNA seq) is its probing capacity to detect deranged or abnormal cell populations present within homogenously perceived pooled cells, which would have evaded cursory screening on the pooled cell populations of biological samples obtained as part of diagnostic procedures. Despite conduction of just single-cell transcriptome analysis, scRNAseq now permits comparison of the transcriptome of the individual cells, which can be evaluated for gene expressional patterns that depict areas of heterogeneity with pharmaceutical drug discovery and clinical treatment applications. It is vital to strictly work through the tools of investigations from wet lab to bioinformatics and computational tooled analyses. In the precise steps for scRNAseq, it is critical to do thorough and effective isolation of viable single cells from the tissues of interest using dependable techniques (such as FACS) before proceeding to lysis, as this enhances the appropriate picking of quality mRNA molecules for subsequent sequencing (such as by the use of Polymerase Chain Reaction machine). Interestingly, scRNAseq can be deployed to analyze various types of biological samples such as embryos, nervous systems, tumour cells, stem cells, lymphocytes, and haematopoietic cells. In haematopoietic cells, it can be used to stratify acute myeloid leukemia patterns in patients, sorting them out into cohorts that enable re-modeling of treatment regimens based on stratified presentations. In immunotherapy, it can furnish specialist clinician-immunologist with tools to re-model treatment for each patient, an attribute of precision medicine. Finally, the good predictive attribute of scRNAseq can help reduce the cost of treatment for patients, thus attracting more patients who would have otherwise been discouraged from seeking quality clinical consultation help due to perceived high cost. This is a positive paradigm shift for patients’ attitudes primed towards seeking treatment.

Keywords: immunotherapy, transcriptome, re-modeling, mRNA, scRNA-seq

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Authors: Trina Isabel D. Santiago, Juan Raphael M. Perez, Maria Angelica O. Soriano, Teresita B. Suing, Jumee F. Tayaban

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To achieve universal healthcare for everyone, the World Health Organization has emphasized the importance of National Medicines Policies for increased accessibility and utilization of high-quality and affordable medications. In the Philippines, significant challenges have been identified surrounding the sustainability of essential medicines, resulting in limited access such as high cost and dominance and market dominance and monopoly of multinational companies (MNCs) in the Philippine pharmaceutical industry. These identified challenges have been addressed by several initiatives, such as the Philippine National Drug Policy and Generics Act of 1988 (Republic Act 6675), to attempt to reduce drug prices. Despite these efforts, the concerns with drug accessibility and affordability continue to persist; hence, Republic Act 9502 was enacted. This paper attempts to review RA 9502 in the pursuit of making medicines more affordable for Filipinos, analyze and critique the problems and challenges associated with the law, and provide recommendations to address identified problems and challenges. A literature search and review, as well as an analysis of the law, has been done to evaluate the policy. RA 9502 recognizes the importance of market competition in drug price reduction and quality medicine accessibility. Contentious issues prior to enactment of the law include 1) parallel importation, pointing out that the drug price will depend on the global market price, 2) contrasting approaches in the drafting of the law as the House version focused on medicine price control while the Senate version prioritized market competition, and 3) MNCs opposing the amendments with concerns on discrimination, constitutional violations, and noncompliance with international treaty obligations. There are also criticisms and challenges with the implementation of the law in terms of content or modeling, interpretation and implementation, and other external factors or hindrances. The law has been criticized for its narrow scope as it only covers specific essential medicines with no cooperation with the national health insurance program. Moreover, the law has sections taking advantage of the TRIPS flexibilities, which disallow smaller countries to reap the benefits of flexibilities. The sanctions and penalties have an insignificant role in implementation as they only ask for a small portion of the income of MNCs. Proposed recommendations for policy improvement include aligning existing legislation through strengthened price regulation and expanded law coverage, strengthening penalties to promote law adherence, and promoting research and development to encourage and support local initiatives. Through these comprehensive recommendations, the issues surrounding the policy can be addressed, and the goal of enhancing the affordability and accessibility of medicines in the country can be achieved.

Keywords: drug accessibility, drug affordability, price regulation, Republic Act 9502

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Authors: Razieh Karimi Aghcheh, Christian Kubicek, Joseph Strauss, Gerhard Braus

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Filamentous fungi are of considerable economic and social significance for human health, nutrition and in white biotechnology. These organisms are dominant producers of a range of primary metabolites such as citric acid, microbial lipids (biodiesel) and higher unsaturated fatty acids (HUFAs). In particular, they produce also important but structurally complex secondary metabolites with enormous therapeutic applications in pharmaceutical industry, for example: cephalosporin, penicillin, taxol, zeranol and ergot alkaloids. Several fungal secondary metabolites, which are significantly relevant to human health do not only include antibiotics, but also e.g. lovastatin, a well-known antihypercholesterolemic agent produced by Aspergillus. terreus, or aflatoxin, a carcinogen produced by A. flavus. In addition to their roles for human health and agriculture, some fungi are industrially and commercially important: Species of the ascomycete genus Hypocrea spp. (teleomorph of Trichoderma) have been demonstrated as efficient producer of highly active cellulolytic enzymes. This trait makes them effective in disrupting and depolymerization of lignocellulosic materials and thus applicable tools in number of biotechnological areas as diverse as clothes-washing detergent, animal feed, and pulp and fuel productions. Fungal LaeA/LAE1 (Loss of aflR Expression A) homologs their gene products act at the interphase between secondary metabolisms, cellulase production and development. Lack of the corresponding genes results in significant physiological changes including loss of secondary metabolite and lignocellulose degrading enzymes production. At the molecular level, the encoded proteins are presumably methyltransferases or demethylases which act directly or indirectly at heterochromatin and interact with velvet domain proteins. Velvet proteins bind to DNA and affect expression of secondary metabolites (SMs) genes and cellulases. The dynamic interplay between LaeA/LAE1, velvet proteins and additional interaction partners is the key for an understanding of the coordination of metabolic and morphological functions of fungi and is required for a biotechnological control of the formation of desired bioactive products. Aspergilli and Trichoderma represent different biotechnologically significant species with significant differences in the LaeA/LAE1-Velvet protein machinery and their target proteins. We, therefore, performed a comparative study of the interaction partners of this machinery and the dynamics of the various protein-protein interactions using our robust proteomic and mass spectrometry techniques. This enhances our knowledge about the fungal coordination of secondary metabolism, cellulase production and development and thereby will certainly improve recombinant fungal strain construction for the production of industrial secondary metabolite or lignocellulose hydrolytic enzymes.

Keywords: cellulases, LaeA/1, proteomics, secondary metabolites

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Authors: Bensaid A., Mostephaoui T., Nedjai R.

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A considerable area of Algerian lands are threatened by the phenomenon of wind erosion. For a long time, wind erosion and its associated harmful effects on the natural environment have posed a serious threat, especially in the arid regions of the country. In recent years, as a result of increases in the irrational exploitation of natural resources (fodder) and extensive land clearing, wind erosion has particularly accentuated. The extent of degradation in the arid region of the Algerian Mécheriadepartment generated a new situation characterized by the reduction of vegetation cover, the decrease of land productivity, as well as sand encroachment on urban development zones. In this study, we attempt to investigate the potential of remote sensing and geographic information systems for detecting the spatial dynamics of the ancient dune cords based on the numerical processing of PlanetScope PSB.SB sensors images by September 29, 2021. As a second step, we prospect the use of a multi-scale geographic object-based image analysis (GEOBIA) approach to segment the high spatial resolution images acquired on heterogeneous surfaces that vary according to human influence on the environment. We have used the fractal net evolution approach (FNEA) algorithm to segment images (Baatz&Schäpe, 2000). Multispectral data, a digital terrain model layer, ground truth data, a normalized difference vegetation index (NDVI) layer, and a first-order texture (entropy) layer were used to segment the multispectral images at three segmentation scales, with an emphasis on accurately delineating the boundaries and components of the sand accumulation areas (Dune, dunes fields, nebka, and barkhane). It is important to note that each auxiliary data contributed to improve the segmentation at different scales. The silted areas were classified using a nearest neighbor approach over the Naâma area using imagery. The classification of silted areas was successfully achieved over all study areas with an accuracy greater than 85%, although the results suggest that, overall, a higher degree of landscape heterogeneity may have a negative effect on segmentation and classification. Some areas suffered from the greatest over-segmentation and lowest mapping accuracy (Kappa: 0.79), which was partially attributed to confounding a greater proportion of mixed siltation classes from both sandy areas and bare ground patches. This research has demonstrated a technique based on very high-resolution images for mapping sanded and degraded areas using GEOBIA, which can be applied to the study of other lands in the steppe areas of the northern countries of the African continent.

Keywords: land development, GIS, sand dunes, segmentation, remote sensing

Procedia PDF Downloads 109

Authors: Manish Saha, Manish Kumar Niranjan, Saket Asthana

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The K₀.₅Na₀.₅NbO₃ (KNN) system has emerged as one of the most promising lead-free piezoelectric over the years. In this work, we perform a comprehensive investigation of electronic structure, lattice dynamics and dielectric/ferroelectric properties of the room temperature phase of KNN by combining ab-initio DFT-based theoretical analysis and experimental characterization. We assign the symmetry labels to KNN vibrational modes and obtain ab-initio polarized Raman spectra, Infrared (IR) reflectivity, Born-effective charge tensors, oscillator strengths etc. The computed Raman spectrum is found to agree well with the experimental spectrum. In particular, the results suggest that the mode in the range ~840-870 cm-¹ reported in the experimental studies is longitudinal optical (LO) with A_1 symmetry. The Raman mode intensities are calculated for different light polarization set-ups, which suggests the observation of different symmetry modes in different polarization set-ups. The electronic structure of KNN is investigated, and an optical absorption spectrum is obtained. Further, the performances of DFT semi-local, metal-GGA and hybrid exchange-correlations (XC) functionals, in the estimation of KNN band gaps are investigated. The KNN bandgap computed using GGA-1/2 and HSE06 hybrid functional schemes are found to be in excellant agreement with the experimental value. The COHP, electron localization function and Bader charge analysis is also performed to deduce the nature of chemical bonding in the KNN. The solid-state reaction and hydrothermal methods are used to prepare the KNN ceramics, and the effects of grain size on the physical characteristics these ceramics are examined. A comprehensive study on the impact of different synthesis techniques on the structural, electrical, and photocatalytic properties of ferroelectric ceramics KNN. The KNN-S prepared by solid-state method have significantly larger grain size as compared to that for KNN-H prepared by hydrothermal method. Furthermore, the KNN-S is found to exhibit higher dielectric, piezoelectric and ferroelectric properties as compared to KNN-H. On the other hand, the increased photocatalytic activity is observed in KNN-H as compared to KNN-S. As compared to the hydrothermal synthesis, the solid-state synthesis causes an increase in the relative dielectric permittivity (ε^') from 2394 to 3286, remnant polarization (P_r) from 15.38 to 20.41 μC/cm^², planer electromechanical coupling factor (k_p) from 0.19 to 0.28 and piezoelectric coefficient (d_33) from 88 to 125 pC/N. The KNN-S ceramics are also found to have a lower leakage current density, and higher grain resistance than KNN-H ceramic. The enhanced photocatalytic activity of KNN-H is attributed to relatively smaller particle sizes. The KNN-S and KNN-H samples are found to have degradation efficiencies of RhB solution of 20% and 65%, respectively. The experimental study highlights the importance of synthesis methods and how these can be exploited to tailor the dielectric, piezoelectric and photocatalytic properties of KNN. Overall, our study provides several bench-mark important results on KNN that have not been reported so far.

Keywords: lead-free piezoelectric, Raman intensity spectrum, electronic structure, first-principles calculations, solid state synthesis, photocatalysis, hydrothermal synthesis

Procedia PDF Downloads 49

Authors: Maja Benkovic, Dubravka Novotni, Bojana Voucko, Duska Curic, Damir Jezek, Nikolina Cukelj

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Cryo-grinding is an ultra-fine grinding method used in the pharmaceutical industry, production of herbs and spices and in the production and handling of cereals, due to its ability to produce powders with small particle sizes which maintain their favorable bioactive profile. The aim of this study was to determine the particle size distributions of the proso millet (Panicum miliaceum) bran fraction grinded at cryogenic temperature (using liquid nitrogen (LN₂) cooling, T = - 196 °C), in comparison to non-cooled grinding. Proso millet bran is primarily used as an animal feed, but has a potential in food applications, either as a substrate for extraction of bioactive compounds or raw material in the bakery industry. For both applications finer particle sizes of the bran could be beneficial. Thus, millet bran was ground for 2, 4, 8 and 12 minutes using the ball mill (CryoMill, Retsch GmbH, Haan, Germany) at three grinding modes: (I) without cooling, (II) at cryo-temperature, and (III) at cryo-temperature with included 1 minute of intermediate cryo-cooling step after every 2 minutes of grinding, which is usually applied when samples require longer grinding times. The sample was placed in a 50 mL stainless steel jar containing one grinding ball (Ø 25 mm). The oscillation frequency in all three modes was 30 Hz. Particle size distributions of the bran were determined by a laser diffraction particle sizing method (Mastersizer 2000) using the Scirocco 2000 dry dispersion unit (Malvern Instruments, Malvern, UK). Three main effects of the grinding set-up were visible from the results. Firstly, grinding time at all three modes had a significant effect on all particle size parameters: d(0.1), d(0.5), d(0.9), D[3,2], D[4,3], span and specific surface area. Longer grinding times resulted in lower values of the above-listed parameters, e.g. the averaged d(0.5) of the sample (229.57±1.46 µm) dropped to 51.29±1.28 µm after 2 minutes grinding without LN₂, and additionally to 43.00±1.33 µm after 4 minutes of grinding without LN₂. The only exception was the sample ground for 12 minutes without cooling, where an increase in particle diameters occurred (d(0.5)=62.85±2.20 µm), probably due to particles adhering to one another and forming larger particle clusters. Secondly, samples with LN₂ cooling exhibited lower diameters in comparison to non-cooled. For example, after 8 minutes of non-cooled grinding d(0.5)=46.97±1.05 µm was achieved, while the LN₂ cooling enabled collection of particles with average sizes of d(0.5)=18.57±0.18 µm. Thirdly, the application of intermediate cryo-cooling step resulted in similar particle diameters (d(0.5)=15.83±0.36 µm, 12 min of grinding) as cryo-milling without this step (d(0.5)=16.33±2.09 µm, 12 min of grinding). This indicates that intermediate cooling is not necessary for the current application, which consequently reduces the consumption of LN₂. These results point out the potential beneficial effects of millet bran grinding at cryo-temperatures. Further research will show if the lower particle size achieved in comparison to non-cooled grinding could result in increased bioavailability of bioactive compounds, as well as protein digestibility and solubility of dietary fibers of the proso millet bran fraction.

Keywords: ball mill, cryo-milling, particle size distribution, proso millet (Panicum miliaceum) bran

Procedia PDF Downloads 145

Authors: Hongmei Wang, Ziyun Xiang, Ying liu, Li Yu, Dongsheng Yue

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Intro: The COVID-19 (Corona Virus Disease 2019) has greatly changed the global economic, political and financial ecology. The mutation of the coronavirus in the UK in December 2020 has brought new panic to the world. Deep learning was performed on Chest Computed tomography (CT) of COVID-19 and Influenza and describes their characteristics. The predominant features of COVID-19 pneumonia was ground-glass opacification, followed by consolidation. Lesion density: most lesions appear as ground-glass shadows, and some lesions coexist with solid lesions. Lesion distribution: the focus is mainly on the dorsal side of the periphery of the lung, with the lower lobe of the lungs as the focus, and it is often close to the pleura. Other features it has are grid-like shadows in ground glass lesions, thickening signs of diseased vessels, air bronchi signs and halo signs. The severe disease involves whole bilateral lungs, showing white lung signs, air bronchograms can be seen, and there can be a small amount of pleural effusion in the bilateral chest cavity. At the same time, this year's flu season could be near its peak after surging throughout the United States for months. Chest CT for Influenza infection is characterized by focal ground glass shadows in the lungs, with or without patchy consolidation, and bronchiole air bronchograms are visible in the concentration. There are patchy ground-glass shadows, consolidation, air bronchus signs, mosaic lung perfusion, etc. The lesions are mostly fused, which is prominent near the hilar and two lungs. Grid-like shadows and small patchy ground-glass shadows are visible. Deep neural networks have great potential in image analysis and diagnosis that traditional machine learning algorithms do not. Method: Aiming at the two major infectious diseases COVID-19 and influenza, which are currently circulating in the world, the chest CT of patients with two infectious diseases is classified and diagnosed using deep learning algorithms. The residual network is proposed to solve the problem of network degradation when there are too many hidden layers in a deep neural network (DNN). The proposed deep residual system (ResNet) is a milestone in the history of the Convolutional neural network (CNN) images, which solves the problem of difficult training of deep CNN models. Many visual tasks can get excellent results through fine-tuning ResNet. The pre-trained convolutional neural network ResNet is introduced as a feature extractor, eliminating the need to design complex models and time-consuming training. Fastai is based on Pytorch, packaging best practices for in-depth learning strategies, and finding the best way to handle diagnoses issues. Based on the one-cycle approach of the Fastai algorithm, the classification diagnosis of lung CT for two infectious diseases is realized, and a higher recognition rate is obtained. Results: A deep learning model was developed to efficiently identify the differences between COVID-19 and influenza using chest CT.

Keywords: COVID-19, Fastai, influenza, transfer network

Procedia PDF Downloads 142

Authors: Chandan Kundu, Sankar Bhattacharya

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A significant disadvantage of fossil fuel energy production is the considerable amount of carbon dioxide (CO₂) released, which is one of the contributors to climate change. Apart from environmental concerns, changing fossil fuel prices have pushed society gradually towards renewable energy sources in recent years. Biomass is a plentiful and renewable resource and a source of carbon. Recent years have seen increased research interest in generating fuels and chemicals from biomass. Unlike fossil-based resources, biomass is composed of lignocellulosic material, which does not contribute to the increase in atmospheric CO₂ over a longer term. These considerations contribute to the current move of the chemical industry from non-renewable feedstock to renewable biomass. This presentation focuses on generating bio-oil and two major platform chemicals that can potentially improve the environment. Thermochemical processes such as pyrolysis are considered viable methods for producing bio-oil and biomass-based platform chemicals. Fluidized bed reactors, on the other hand, are known to boost bio-oil yields during pyrolysis due to their superior mixing and heat transfer features, as well as their scalability. This review and the associated experimental work are focused on the thermochemical conversion of biomass to bio-oil and two high-value platform chemicals, Levoglucosenone (LGO) and 5-Chloromethyl furfural (5-CMF), in a fluidized bed reactor. These two active molecules with distinct features can potentially be useful monomers in the chemical and pharmaceutical industries since they are well adapted to the manufacture of biologically active products. This process took several meticulous steps. To begin, the biomass was delignified using a peracetic acid pretreatment to remove lignin. Because of its complicated structure, biomass must be pretreated to remove the lignin, increasing access to the carbohydrate components and converting them to platform chemicals. The biomass was then characterized by Thermogravimetric analysis, Synchrotron-based THz spectroscopy, and in-situ DRIFTS in the laboratory. Based on the results, a continuous-feeding fluidized bed reactor system was constructed to generate platform chemicals from pretreated biomass using hydrogen chloride acid-gas as a catalyst. The procedure also yields biochar, which has a number of potential applications, including soil remediation, wastewater treatment, electrode production, and energy resource utilization. Consequently, this research also includes a preliminary experimental evaluation of the biochar's prospective applications. The biochar obtained was evaluated for its CO₂ and steam reactivity. The outline of the presentation will comprise the following: Biomass pretreatment for effective delignification Mechanistic study of the thermal and thermochemical conversion of biomass Thermochemical conversion of untreated and pretreated biomass in the presence of an acid catalyst to produce LGO and CMF A thermo-catalytic process for the production of LGO and 5-CMF in a continuously-fed fluidized bed reactor and efficient separation of chemicals Use of biochar generated from the platform chemicals production through gasification

Keywords: biomass, pretreatment, pyrolysis, levoglucosenone

Procedia PDF Downloads 141

Authors: Marita Pigłowska, Beata Kurc, Maciej Galiński

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The application of carbon materials in the branches of the electrochemical industry shows an increasing tendency each year due to the many interesting properties they possess. These are, among others, a well-developed specific surface, porosity, high sorption capacity, good adsorption properties, low bulk density, electrical conductivity and chemical resistance. All these properties allow for their effective use, among others in supercapacitors, which can store electric charges of the order of 100 F due to carbon electrodes constituting the capacitor plates. Coals (including expanded graphite, carbon black, graphite carbon fibers, activated carbon) are commonly used in electrochemical methods of removing oil derivatives from water after tanker disasters, e.g. phenols and their derivatives by their electrochemical anodic oxidation. Phenol can occupy practically the entire surface of carbon material and leave the water clean of hydrophobic impurities. Regeneration of such electrodes is also not complicated, it is carried out by electrochemical methods consisting in unblocking the pores and reducing resistances, and thus their reactivation for subsequent adsorption processes. Graphite is commonly used as an anode material in lithium-ion cells, while due to the limited capacity it offers (372 mAh g-1), new solutions are sought that meet both capacitive, efficiency and economic criteria. Increasingly, biodegradable materials, green materials, biomass, waste (including agricultural waste) are used in order to reuse them and reduce greenhouse effects and, above all, to meet the biodegradability criterion necessary for the production of lithium-ion cells as chemical power sources. The most common of these materials are cellulose, starch, wheat, rice, and corn waste, e.g. from agricultural, paper and pharmaceutical production. Such products are subjected to appropriate treatments depending on the desired application (including chemical, thermal, electrochemical). Starch is a biodegradable polysaccharide that consists of polymeric units such as amylose and amylopectin that build an ordered (linear) and amorphous (branched) structure of the polymer. Carbon is also used as a catalyst. Elemental carbon has become available in many nano-structured forms representing the hybridization combinations found in the primary carbon allotropes, and the materials can be enriched with a large number of surface functional groups. There are many examples of catalytic applications of coal in the literature, but the development of this field has been hampered by the lack of a conceptual approach combining structure and function and a lack of understanding of material synthesis. In the context of catalytic applications, the integrity of carbon environmental management properties and parameters such as metal conductivity range and bond sequence management should be characterized. Such data, along with surface and textured information, can form the basis for the provision of network support services.

Keywords: carbon materials, catalysis, BET, capacitors, lithium ion cell

Procedia PDF Downloads 174

Authors: Leslie Raphael de M. Ferraz, Salvana Priscylla M. Costa, Tarcyla de A. Gomes, Giovanna Christinne R. M. Schver, Cristóvão R. da Silva, Magaly Andreza M. de Lyra, Danilo Augusto F. Fontes, Larissa A. Rolim, Amanda Carla Q. M. Vieira, Miracy M. de Albuquerque, Pedro J. Rolim-Neto

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Efavirenz (EFV) is a drug used as first-line treatment of AIDS. However, it has poor aqueous solubility and wettability, presenting problems in the gastrointestinal tract absorption and bioavailability. One of the most promising strategies to improve the solubility is the use of solid dispersions (SD). Therefore, this study aimed to characterize SD EFZ with the polymers: PVP-K30, PVPVA 64 and SOLUPLUS in order to find an optimal formulation to compose a future pharmaceutical product for AIDS therapy. Initially, Physical Mixtures (PM) and SD with the polymers were obtained containing 10, 20, 50 and 80% of drug (w/w) by the solvent method. The best formulation obtained between the SD was selected by in vitro dissolution test. Finally, the drug-carrier system chosen, in all ratios obtained, were analyzed by the following techniques: Differential Scanning Calorimetry (DSC), polarization microscopy, Scanning Electron Microscopy (SEM) and spectrophotometry of absorption in the region of infrared (IR). From the dissolution profiles of EFV, PM and SD, the values of area Under The Curve (AUC) were calculated. The data showed that the AUC of all PM is greater than the isolated EFV, this result is derived from the hydrophilic properties of the polymers thus favoring a decrease in surface tension between the drug and the dissolution medium. In adittion, this ensures an increasing of wettability of the drug. In parallel, it was found that SD whom had higher AUC values, were those who have the greatest amount of polymer (with only 10% drug). As the amount of drug increases, it was noticed that these results either decrease or are statistically similar. The AUC values of the SD using the three different polymers, followed this decreasing order: SD PVPVA 64-EFV 10% > SD PVP-K30-EFV 10% > SD Soluplus®-EFV 10%. The DSC curves of SD’s did not show the characteristic endothermic event of drug melt process, suggesting that the EFV was converted to its amorphous state. The analysis of polarized light microscopy showed significant birefringence of the PM’s, but this was not observed in films of SD’s, thus suggesting the conversion of the drug from the crystalline to the amorphous state. In electron micrographs of all PM, independently of the percentage of the drug, the crystal structure of EFV was clearly detectable. Moreover, electron micrographs of the SD with the two polymers in different ratios investigated, we observed the presence of particles with irregular size and morphology, also occurring an extensive change in the appearance of the polymer, not being possible to differentiate the two components. IR spectra of PM corresponds to the overlapping of polymer and EFV bands indicating thereby that there is no interaction between them, unlike the spectra of all SD that showed complete disappearance of the band related to the axial deformation of the NH group of EFV. Therefore, this study was able to obtain a suitable formulation to overcome the solubility limitations of the EFV, since SD PVPVA 64-EFZ 10% was chosen as the best system in delay crystallization of the prototype, reaching higher levels of super saturation.

Keywords: characterization, dissolution, Efavirenz, solid dispersions

Procedia PDF Downloads 631

Authors: Colin S. Chen, Chien-Jung Tien, Hsin-Jan Huang

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For weathering studies, the change of chemical constituents by biodegradation effect in diesel-contaminated soils are important factors to be considered, especially when there is a prolonged period of weathering processes. The objective was to evaluate biodegradation effects onto hydrocarbon fingerprinting and distribution patterns of diesel fuels, fuel source screening and differentiation, source-specific marker compounds, and diagnostic ratios of diesel fuel constituents by laboratory and field studies. Biodegradation processes of diesel contaminated soils were evaluated by experiments lasting for 15 and 12 months, respectively. The degradation of diesel fuel in top soils was affected by organic carbon content and biomass of microorganisms in soils. Higher depletion of total petroleum hydrocarbon (TPH), n-alkanes, and polynuclear aromatic hydrocarbons (PAHs) and their alkyl homologues was observed in soils containing higher organic carbon content and biomass. Decreased ratio of selected isoprenoids (i.e., pristane (Pr) and phytane (Ph)) including n-C17/pristane and n-C18/phytane was observed. The ratio of pristane/phytane was remained consistent for a longer period of time. At the end of the experimental period, a decrease of pristane/phytane was observed. Biomarker compounds of bicyclic sesquiterpanes (BS) were less susceptible to the effects of biodegradation. The ratios of characteristic factors such as C15 sesquiterpane/ 8β(H)-drimane (BS3/BS5), C15 sesquiterpane/ 8β(H)-drimane (BS4/BS5), 8β(H)-drimane/8β(H)-homodrimane (BS5/BS10), and C15 sesquiterpane/8β(H)-homodrimane (BS3/BS10) could be adopted for source identification of diesel fuels in top soil. However, for biodegradation processes lasted for six months but shorter than nine months, only BS3/BS5 and BS3/BS10 could be distinguished in two diesel fuels. In subsoil experiments (contaminated soil located 50 cm below), the ratios of characteristic factors including BS3/BS5, BS4/BS5, and BS5/BS10 were valid for source identification of two diesel fuels for nine month biodegradation. At the early stage of contamination, biomass of soil decreased significantly. However, 6 and 7 dominant species were found in soils in top soil experiments, respectively. With less oxygen and nutrients in subsoil, less biomass of microorganisms was observed in subsoils. Only 2 and 4 diesel-degrading species of microorganisms were identified in two soils, respectively. Parameters of double ratio such as fluorene/C1-fluorene: C2-phenanthrene/C3-phenanthrene (C0F/C1F:C2P/C3P) in both top and subsoil, C2-naphthalene/C2-phenanthrene: C1-phenanthrene/C3-phenanthrene (C2N/C2P:C1P/C3P), and C1-phenanthrene/C1-fluorene: C3-naphthalene/C3-phenanthrene (C1P/C1F:C3N/C3P) in subsoil could serve as forensic indicators in diesel contaminated sites. BS3/BS10:BS4/BS5 could be used in 6 to 9 months of biodegradation processes. Results of principal component analysis (PCA) indicated that source identification of diesel fuels in top soil could only be perofrmed for weathering process less than 6 months. For subsoil, identification can be conducted for weathering process less than 9 months. Ratio of isoprenoids (pristane and phytane) and PAHs might be affected by biodegradation in spilled sites. The ratios of bicyclic sesquiterpanes could serve as forensic indicators in diesel-contaminated soils. Finally, source identification was attemped for samples collected from different fuel contaminated sites by using the unique pattern of sesquiterpanes. It was anticipated that the information generated from this study would be adopted by decision makers to evaluate the liability of cleanup in diesel contaminated sites.

Keywords: biodegradation, diagnostic ratio, diesel fuel, environmental forensics

Procedia PDF Downloads 228

Authors: G. M. Vlasceanu, H. Iovu, E. Vasile, M. Ionita

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Herein, the synthesis and characterization of chitosan-gelatin highly porous scaffold reinforced with graphene oxide, and hydroxyapatite (HAp), crosslinked with genipin was targeted. In tissue engineering, chitosan and gelatin are two of the most robust biopolymers with wide applicability due to intrinsic biocompatibility, biodegradability, low antigenicity properties, affordability, and ease of processing. HAp, per its exceptional activity in tuning cell-matrix interactions, is acknowledged for its capability of sustaining cellular proliferation by promoting bone-like native micro-media for cell adjustment. Genipin is regarded as a top class cross-linker, while graphene oxide (GO) is viewed as one of the most performant and versatile fillers. The composites with natural bone HAp/biopolymer ratio were obtained by cascading sonochemical treatments, followed by uncomplicated casting methods and by freeze-drying. Their structure was characterized by Fourier Transform Infrared Spectroscopy and X-ray Diffraction, while overall morphology was investigated by Scanning Electron Microscopy (SEM) and micro-Computer Tomography (µ-CT). Ensuing that, in vitro enzyme degradation was performed to detect the most promising compositions for the development of in vivo assays. Suitable GO dispersion was ascertained within the biopolymer mix as nanolayers specific signals lack in both FTIR and XRD spectra, and the specific spectral features of the polymers persisted with GO load enhancement. Overall, correlations between the GO induced material structuration, crystallinity variations, and chemical interaction of the compounds can be correlated with the physical features and bioactivity of each composite formulation. Moreover, the HAp distribution within follows an auspicious density gradient tuned for hybrid osseous/cartilage matter architectures, which were mirrored in the mice model tests. Hence, the synthesis route of a natural polymer blend/hydroxyapatite-graphene oxide composite material is anticipated to emerge as influential formulation in bone tissue engineering. Acknowledgement: This work was supported by the project 'Work-based learning systems using entrepreneurship grants for doctoral and post-doctoral students' (Sisteme de invatare bazate pe munca prin burse antreprenor pentru doctoranzi si postdoctoranzi) - SIMBA, SMIS code 124705 and by a grant of the National Authority for Scientific Research and Innovation, Operational Program Competitiveness Axis 1 - Section E, Program co-financed from European Regional Development Fund 'Investments for your future' under the project number 154/25.11.2016, P_37_221/2015. The nano-CT experiments were possible due to European Regional Development Fund through Competitiveness Operational Program 2014-2020, Priority axis 1, ID P_36_611, MySMIS code 107066, INOVABIOMED.

Keywords: biopolymer blend, ectopic osteoinduction, graphene oxide composite, hydroxyapatite

Procedia PDF Downloads 104

Authors: Marcin Zieliński, Marcin Dębowski, Mirosław Krzemieniewski

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The anaerobic degradation of substrates is limited especially by the rate and effectiveness of the first (hydrolytic) stage of fermentation. This stage may be intensified through pre-treatment of substrate aimed at disintegration of the solid phase and destruction of substrate tissues and cells. The most frequently applied criterion of disintegration outcomes evaluation is the increase in biogas recovery owing to the possibility of its use for energetic purposes and, simultaneously, recovery of input energy consumed for the pre-treatment of substrate before fermentation. Hydrodynamic cavitation is one of the methods for organic substrate disintegration that has a high implementation potential. Cavitation is explained as the phenomenon of the formation of discontinuity cavities filled with vapor or gas in a liquid induced by pressure drop to the critical value. It is induced by a varying field of pressures. A void needs to occur in the flow in which the pressure first drops to the value close to the pressure of saturated vapor and then increases. The process of cavitation conducted under controlled conditions was found to significantly improve the effectiveness of anaerobic conversion of organic substrates having various characteristics. This phenomenon allows effective damage and disintegration of cellular and tissue structures. Disintegration of structures and release of organic compounds to the dissolved phase has a direct effect on the intensification of biogas production in the process of anaerobic fermentation, on reduced dry matter content in the post-fermentation sludge as well as a high degree of its hygienization and its increased susceptibility to dehydration. A device the efficiency of which was confirmed both in laboratory conditions and in systems operating in the technical scale is a hydrodynamic generator of cavitation. Cavitators, agitators and emulsifiers constructed and tested worldwide so far have been characterized by low efficiency and high energy demand. Many of them proved effective under laboratory conditions but failed under industrial ones. The only task successfully realized by these appliances and utilized on a wider scale is the heating of liquids. For this reason, their usability was limited to the function of heating installations. Design of the presented cavitation generator allows achieving satisfactory energy efficiency and enables its use under industrial conditions in depolymerization processes of biomass with various characteristics. Investigations conducted on the laboratory and industrial scale confirmed the effectiveness of applying cavitation in the process of biomass destruction. The use of the cavitation generator in laboratory studies for disintegration of sewage sludge allowed increasing biogas production by ca. 30% and shortening the treatment process by ca. 20 - 25%. The shortening of the technological process and increase of wastewater treatment plant effectiveness may delay investments aimed at increasing system output. The use of a mechanical cavitator and application of repeated cavitation process (4-6 times) enables significant acceleration of the biogassing process. In addition, mechanical cavitation accelerates increases in COD and VFA levels.

Keywords: hydrodynamic cavitation, pretreatment, biomass, methane fermentation, Virginia fanpetals

Procedia PDF Downloads 434

Authors: T. Ghatauray, J. Ingram, P. Holborn

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The transition from a ‘carbon rich’ fossil fuel dependent to a ‘sustainable’ and ‘renewable’ hydrogen based society will see the deployment of hydrogen fuel cells (HFC) in transport applications and in the generation of heat and power for buildings, as part of a decentralised power network. Many deployments will be low power HFCs for domestic combined heat and power (CHP) and commercial ‘transportable’ HFCs for environmental situations, such as lighting and telephone towers. For broad commercialisation of small fuel cells to be achieved there needs to be significant confidence in their safety in both domestic and environmental applications. Low power HFCs are housed in protective steel enclosures. Standard enclosures have plain rectangular ventilation openings intended for thermal management of electronics and not the dispersion of a buoyant gas. Degradation of the HFC or supply pipework in use could lead to a low-level leak and a build-up of hydrogen gas in the enclosure. Hydrogen’s wide flammable range (4-75%) is a significant safety concern, with ineffective enclosure ventilation having the potential to cause flammable mixtures to develop with the risk of explosion. Mechanical ventilation is effective at managing enclosure hydrogen concentrations, but drains HFC power and is vulnerable to failure. This is undesirable in low power and remote installations and reliable passive ventilation systems are preferred. Passive ventilation depends upon buoyancy driven flow, with the size, shape and position of ventilation openings critical for producing predictable flows and maintaining low buoyant gas concentrations. With environmentally sited enclosures, ventilation openings with pressed horizontal and angled louvres are preferred to protect the HFC and electronics inside. There is an economic cost to adding louvres, but also a safety concern. A question arises over whether the use of pressed louvre vents impairs enclosure passive ventilation performance, when compared to same opening area plain vents. Comparison small enclosure (0.144m³) tests of same opening area pressed louvre and plain vents were undertaken. A displacement ventilation arrangement was incorporated into the enclosure with opposing upper and lower ventilation openings. A range of vent areas were tested. Helium (used as a safe analogue for hydrogen) was released from a 4mm nozzle at the base of the enclosure to simulate a hydrogen leak at leak rates from 1 to 10 lpm. Helium sensors were used to record concentrations at eight heights in the enclosure. The enclosure was otherwise empty. These tests determined that the use of pressed and angled louvre ventilation openings on the enclosure impaired the passive ventilation flow and increased helium concentrations in the enclosure. High-level stratified buoyant gas layers were also found to be deeper than with plain vent openings and were within the flammable range. The presence of gas within the flammable range is of concern, particularly as the addition of the fuel cell and electronics in the enclosure would further reduce the available volume and increase concentrations. The opening area of louvre vents would need to be greater than equivalent plain vents to achieve comparable ventilation flows or alternative schemes would need to be considered.

Keywords: enclosure, fuel cell, helium, hydrogen safety, louvre vent, passive ventilation

Procedia PDF Downloads 273

Authors: Darren A. Whitaker, Dan Palmer, Jens Wesholowski, James Flaherty, John Mack, Ahmad B. Albadarin, Gavin Walker

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Traditional mechanosynthesis has been performed by either ball milling or manual grinding. However, neither of these techniques allow the easy application of process control. The temperature may change unpredictably due to friction in the process. Hence the amount of energy transferred to the reactants is intrinsically non-uniform. Recently, it has been shown that the use of Twin-Screw extrusion (TSE) can overcome these limitations. Additionally, TSE enables a platform for continuous synthesis or manufacturing as it is an open-ended process, with feedstocks at one end and product at the other. Several materials including metal-organic frameworks (MOFs), co-crystals and small organic molecules have been produced mechanochemically using TSE. The described advantages of TSE are offset by drawbacks such as increased process complexity (a large number of process parameters) and variation in feedstock flow impacting on product quality. To handle the above-mentioned drawbacks, this study utilizes UV-Vis spectrophotometry (InSpectroX, ColVisTec) as an online tool to gain real-time information about the quality of the product. Additionally, this is combined with real-time process information in an Advanced Process Control system (PharmaMV, Perceptive Engineering) allowing full supervision and control of the TSE process. Further, by characterizing the dynamic behavior of the TSE, a model predictive controller (MPC) can be employed to ensure the process remains under control when perturbed by external disturbances. Two reactions were studied; a Knoevenagel condensation reaction of barbituric acid and vanillin and, the direct amidation of hydroquinone by ammonium acetate to form N-Acetyl-para-aminophenol (APAP) commonly known as paracetamol. Both reactions could be carried out continuously using TSE, nuclear magnetic resonance (NMR) spectroscopy was used to confirm the percentage conversion of starting materials to product. This information was used to construct partial least squares (PLS) calibration models within the PharmaMV development system, which relates the percent conversion to product to the acquired UV-Vis spectrum. Once this was complete, the model was deployed within the PharmaMV Real-Time System to carry out automated optimization experiments to maximize the percentage conversion based on a set of process parameters in a design of experiments (DoE) style methodology. With the optimum set of process parameters established, a series of PRBS process response tests (i.e. Pseudo-Random Binary Sequences) around the optimum were conducted. The resultant dataset was used to build a statistical model and associated MPC. The controller maximizes product quality whilst ensuring the process remains at the optimum even as disturbances such as raw material variability are introduced into the system. To summarize, a combination of online spectral monitoring and advanced process control was used to develop a robust system for optimization and control of two TSE based mechanosynthetic processes.

Keywords: continuous synthesis, pharmaceutical, spectroscopy, advanced process control

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Authors: Dan Kanmegne

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Africa will become the most populated continent by the end of the century, with around 4 billion inhabitants. Food security and climate changes will become continental issues since agricultural practices depend on climate but also contribute to global emissions and land degradation. Agroforestry has been identified as a cost-efficient and reliable strategy to address these two issues. It is defined as the integrated management of trees and crops/animals in the same land unit. Agroforestry provides benefits in terms of goods (fruits, medicine, wood, etc.) and services (windbreaks, fertility, etc.), and is acknowledged to have a great potential for carbon sequestration; therefore it can be integrated into reduction mechanisms of carbon emissions. Particularly in sub-Saharan Africa, the constraint stands in the lack of information about both areas under agroforestry and the characterization (composition, structure, and management) of each agroforestry system at the country level. This study describes and quantifies “what is where?”, earliest to the quantification of carbon stock in different systems. Remote sensing (RS) is the most efficient approach to map such a dynamic technology as agroforestry since it gives relatively adequate and consistent information over a large area at nearly no cost. RS data fulfill the good practice guidelines of the Intergovernmental Panel On Climate Change (IPCC) that is to be used in carbon estimation. Satellite data are getting more and more accessible, and the archives are growing exponentially. To retrieve useful information to support decision-making out of this large amount of data, satellite data needs to be organized so to ensure fast processing, quick accessibility, and ease of use. A new solution is a data cube, which can be understood as a multi-dimensional stack (space, time, data type) of spatially aligned pixels and used for efficient access and analysis. A data cube for Burkina Faso has been set up from the cooperation project between the international service provider WASCAL and Germany, which provides an accessible exploitation architecture of multi-temporal satellite data. The aim of this study is to map and characterize agroforestry systems using the Burkina Faso earth observation data cube. The approach in its initial stage is based on an unsupervised image classification of a normalized difference vegetation index (NDVI) time series from 2010 to 2018, to stratify the country based on the vegetation. Fifteen strata were identified, and four samples per location were randomly assigned to define the sampling units. For safety reasons, the northern part will not be part of the fieldwork. A total of 52 locations will be visited by the end of the dry season in February-March 2020. The field campaigns will consist of identifying and describing different agroforestry systems and qualitative interviews. A multi-temporal supervised image classification will be done with a random forest algorithm, and the field data will be used for both training the algorithm and accuracy assessment. The expected outputs are (i) map(s) of agroforestry dynamics, (ii) characteristics of different systems (main species, management, area, etc.); (iii) assessment report of Burkina Faso data cube.

Keywords: agroforestry systems, Burkina Faso, earth observation data cube, multi-temporal image classification

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Authors: Akram Abi, Mohammad Hossein Sarrafzadeh

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Because of their several advantages over chemical surfactants, biosurfactants have given rise to a growing interest in the past decades. Advantages such as lower toxicity, higher biodegradability, higher selectivity and applicable at extreme temperature and pH which enables them to be used in a variety of applications such as: enhanced oil recovery, environmental and pharmaceutical applications, etc. Bacillus subtilis produces a cyclic lipopeptide, called surfactin, which is one of the most powerful biosurfactants with ability to decrease surface tension of water from 72 mN/m to 27 mN/m. In addition to its biosurfactant character, surfactin exhibits interesting biological activities such as: inhibition of fibrin clot formation, lyses of erythrocytes and several bacterial spheroplasts, antiviral, anti-tumoral and antibacterial properties. Surfactin is an antibiotic substance and has been shown recently to possess anti-HIV activity. However, application of biosurfactants is limited by their high production cost. The cost can be reduced by optimizing biosurfactant production using cheap feed stock. Utilization of inexpensive substrates and unconventional carbon sources like urban or agro-industrial wastes is a promising strategy to decrease the production cost of biosurfactants. With suitable engineering optimization and microbiological modifications, these wastes can be used as substrates for large-scale production of biosurfactants. As an effort to fulfill this purpose, in this work we have tried to utilize olive oil as second carbon source and also yeast extract as second nitrogen source to investigate the effect on both biomass and biosurfactant production improvement in Bacillus subtilis cultures. Since the turbidity of the culture was affected by presence of the oil, optical density was compromised and no longer could be used as an index of growth and biomass concentration. Therefore, cell Dry Weight measurements with applying necessary tactics for removing oil drops to prevent interference with biomass weight were carried out to monitor biomass concentration during the growth of the bacterium. The surface tension and critical micelle dilutions (CMD-1, CMD-2) were considered as an indirect measurement of biosurfactant production. Distinctive and promising results were obtained in the cultures containing olive oil compared to cultures without it: more than two fold increase in biomass production (from 2 g/l to 5 g/l) and considerable reduction in surface tension, down to 40 mN/m at surprisingly early hours of culture time (only 5hr after inoculation). This early onset of biosurfactant production in this culture is specially interesting when compared to the conventional cultures at which this reduction in surface tension is not obtained until 30 hour of culture time. Reducing the production time is a very prominent result to be considered for large scale process development. Furthermore, these results can be used to develop strategies for utilization of agro-industrial wastes (such as olive oil mill residue, molasses, etc.) as cheap and easily accessible feed stocks to decrease the high costs of biosurfactant production.

Keywords: agro-industrial waste, bacillus subtilis, biosurfactant, fermentation, second carbon and nitrogen source, surfactin

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Authors: Hazel James P. Agngarayngay, Arnold R. Elepaño

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Mango is the third most important export fruit in the Philippines. Despite the expanding mango trade in world market, problems on postharvest losses caused by pests and diseases are still prevalent. Many disease control and pest disinfestation methods have been studied and adopted. Heat treatment is necessary to eliminate pests and diseases to be able to pass the quarantine requirements of importing countries. During heat treatments, temperature and time are critical because fruits can easily be damaged by over-exposure to heat. Modeling the process enables researchers and engineers to study the behaviour of temperature distribution within the fruit over time. Understanding physical processes through modeling and simulation also saves time and resources because of reduced experimentation. This research aimed to simulate the heat transfer mechanism and predict the temperature distribution in ‘Carabao' mangoes during hot water treatment (HWT) and extended hot water treatment (EHWT). The simulation was performed in ANSYS CFD Software, using ANSYS CFX Solver. The simulation process involved model creation, mesh generation, defining the physics of the model, solving the problem, and visualizing the results. Boundary conditions consisted of the convective heat transfer coefficient and a constant free stream temperature. The three-dimensional energy equation for transient conditions was numerically solved to obtain heat flux and transient temperature values. The solver utilized finite volume method of discretization. To validate the simulation, actual data were obtained through experiment. The goodness of fit was evaluated using mean temperature difference (MTD). Also, t-test was used to detect significant differences between the data sets. Results showed that the simulations were able to estimate temperatures accurately with MTD of 0.50 and 0.69 °C for the HWT and EHWT, respectively. This indicates good agreement between the simulated and actual temperature values. The data included in the analysis were taken at different locations of probe punctures within the fruit. Moreover, t-tests showed no significant differences between the two data sets. Maximum heat fluxes obtained at the beginning of the treatments were 394.15 and 262.77 J.s-1 for HWT and EHWT, respectively. These values decreased abruptly at the first 10 seconds and gradual decrease was observed thereafter. Data on heat flux is necessary in the design of heaters. If underestimated, the heating component of a certain machine will not be able to provide enough heat required by certain operations. Otherwise, over-estimation will result in wasting of energy and resources. This study demonstrated that the simulation was able to estimate temperatures accurately. Thus, it can be used to evaluate the influence of various treatment conditions on the temperature-time history in mangoes. When combined with information on insect mortality and quality degradation kinetics, it could predict the efficacy of a particular treatment and guide appropriate selection of treatment conditions. The effect of various parameters on heat transfer rates, such as the boundary and initial conditions as well as the thermal properties of the material, can be systematically studied without performing experiments. Furthermore, the use of ANSYS software in modeling and simulation can be explored in modeling various systems and processes.

Keywords: heat transfer, heat treatment, mango, modeling and simulation

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Authors: Manavjit Singh Dhindsa, Ursula Das, Kshirasagar Naik, Marzia Zaman, Richard Purcell, Srinivas Sampalli, Abdul Mutakabbir, Chung-Horng Lung, Thambirajah Ravichandran

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These days, wildland fires, also known as forest fires, are more prevalent than ever. Wildfires have major repercussions that affect ecosystems, communities, and the environment in several ways. Wildfires lead to habitat destruction and biodiversity loss, affecting ecosystems and causing soil erosion. They also contribute to poor air quality by releasing smoke and pollutants that pose health risks, especially for individuals with respiratory conditions. Wildfires can damage infrastructure, disrupt communities, and cause economic losses. The economic impact of firefighting efforts, combined with their direct effects on forestry and agriculture, causes significant financial difficulties for the areas impacted. This research explores different forest fire spread models and presents a comprehensive review of various techniques and methodologies used in the field. A forest fire spread model is a computational or mathematical representation that is used to simulate and predict the behavior of a forest fire. By applying scientific concepts and data from empirical studies, these models attempt to capture the intricate dynamics of how a fire spreads, taking into consideration a variety of factors like weather patterns, topography, fuel types, and environmental conditions. These models assist authorities in understanding and forecasting the potential trajectory and intensity of a wildfire. Emphasizing the need for a comprehensive understanding of wildfire dynamics, this research explores the approaches, assumptions, and findings derived from various models. By using a comparison approach, a critical analysis is provided by identifying patterns, strengths, and weaknesses among these models. The purpose of the survey is to further wildfire research and management techniques. Decision-makers, researchers, and practitioners can benefit from the useful insights that are provided by synthesizing established information. Fire spread models provide insights into potential fire behavior, facilitating authorities to make informed decisions about evacuation activities, allocating resources for fire-fighting efforts, and planning for preventive actions. Wildfire spread models are also useful in post-wildfire mitigation strategies as they help in assessing the fire's severity, determining high-risk regions for post-fire dangers, and forecasting soil erosion trends. The analysis highlights the importance of customized modeling approaches for various circumstances and promotes our understanding of the way forest fires spread. Some of the known models in this field are Rothermel’s wildland fuel model, FARSITE, WRF-SFIRE, FIRETEC, FlamMap, FSPro, cellular automata model, and others. The key characteristics that these models consider include weather (includes factors such as wind speed and direction), topography (includes factors like landscape elevation), and fuel availability (includes factors like types of vegetation) among other factors. The models discussed are physics-based, data-driven, or hybrid models, also utilizing ML techniques like attention-based neural networks to enhance the performance of the model. In order to lessen the destructive effects of forest fires, this initiative aims to promote the development of more precise prediction tools and effective management techniques. The survey expands its scope to address the practical needs of numerous stakeholders. Access to enhanced early warning systems enables decision-makers to take prompt action. Emergency responders benefit from improved resource allocation strategies, strengthening the efficacy of firefighting efforts.

Keywords: artificial intelligence, deep learning, forest fire management, fire risk assessment, fire simulation, machine learning, remote sensing, wildfire modeling

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Authors: Gianmarco Taveri, Ivo Dlouhy

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Geopolymers are well-suited materials to abate CO2 emission coming from the Portland cement production, and then replace them, in the near future, in building and other applications. The cost of production of geopolymers may be seen the only weakness, but the use of wastes as raw materials could provide a valid solution to this problem, as demonstrated by the successful incorporation of fly-ash, a by-product of thermal power plants, and waste glasses. Recycled glass in waste-derived geopolymers was lately employed as a further silica source. In this work we present, for the first time, the introduction of recycled borosilicate glass (BSG). BSG is actually a waste glass, since it derives from dismantled pharmaceutical vials and cannot be reused in the manufacturing of the original articles. Owing to the specific chemical composition (BSG is an ‘alumino-boro-silicate’), it was conceived to provide the key components of zeolitic networks, such as amorphous silica and alumina, as well as boria (B2O3), which may replace Al2O3 and contribute to the polycondensation process. The solid–state MAS NMR spectroscopy was used to assess the extent of boron oxide incorporation in the structure of geopolymers, and to define the degree of networking. FTIR spectroscopy was utilized to define the degree of polymerization and to detect boron bond vibration into the structure. Mechanical performance was tested by means of 3 point bending (flexural strength), chevron notch test (fracture toughness), compression test (compressive strength), micro-indentation test (Vicker’s hardness). Spectroscopy (SEM and Confocal spectroscopy) was performed on the specimens conducted to failure. FTIR showed a characteristic absorption band attributed to the stretching modes of tetrahedral boron ions, whose tetrahedral configuration is compatible to the reaction product of geopolymerization. 27Al NMR and 29Si NMR spectra were instrumental in understanding the extent of the reaction. 11B NMR spectroscopies evidenced a change of the trigonal boron (BO3) inside the BSG in favor of a quasi-total tetrahedral boron configuration (BO4). Thanks to these results, it was inferred that boron is part of the geopolymeric structure, replacing the Si in the network, similarly to the aluminum, and therefore improving the quality of the microstructure, in favor of a more cross-linked network. As expected, the material gained as much as 25% in compressive strength (45 MPa) compared to the literature, whereas no improvements were detected in flexural strength (~ 5 MPa) and superficial hardness (~ 78 HV). The material also exhibited a low fracture toughness (0.35 MPa*m1/2), with a tangible brittleness. SEM micrographies corroborated this behavior, showing a ragged surface, along with several cracks, due to the high presence of porosity and impurities, acting as preferential points for crack initiation. The 3D pattern of the surface fracture, following the confocal spectroscopy, evidenced an irregular crack propagation, whose proclivity was mainly, but not always, to follow the porosity. Hence, the crack initiation and propagation are largely unpredictable.

Keywords: borosilicate glass, characterization, fly-ash, geopolymerization

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Authors: Meiling Yu, Nadia Rouatbi, Khuloud T. Al-Jamal

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Background: Treatment of neurological disorders with modern medical and surgical approaches remains difficult. Gene therapy, allowing the delivery of genetic materials that encodes potential therapeutic molecules, represents an attractive option. The treatment of brain diseases with gene therapy requires the gene-editing tool to be delivered efficiently to the central nervous system. In this study, we explored the efficiency of different delivery routes, namely intravenous (i.v.), intra-cranial (i.c.), and intra-nasal (i.n.), to deliver stable nucleic acid-lipid particles (SNALPs) containing gene-editing tools namely Cas9 mRNA and sgRNA encoding for GFP as a reporter protein. We hypothesise that SNALPs can reach the brain and perform gene-editing to different extents depending on the administration route. Intranasal administration (i.n.) offers an attractive and non-invasive way to access the brain circumventing the blood–brain barrier. Successful delivery of gene-editing tools to the brain offers a great opportunity for therapeutic target validation and nucleic acids therapeutics delivery to improve treatment options for a range of neurodegenerative diseases. In this study, we utilised Rosa26-Cas9 knock-in mice, expressing GFP, to study brain distribution and gene-editing efficiency of SNALPs after i.v.; i.c. and i.n. routes of administration. Methods: Single guide RNA (sgRNA) against GFP has been designed and validated by in vitro nuclease assay. SNALPs were formulated and characterised using dynamic light scattering. The encapsulation efficiency of nucleic acids (NA) was measured by RiboGreen™ assay. SNALPs were incubated in serum to assess their ability to protect NA from degradation. Rosa26-Cas9 knock-in mice were i.v., i.n., or i.c. administered with SNALPs to test in vivo gene-editing (GFP knockout) efficiency. SNALPs were given as three doses of 0.64 mg/kg sgGFP following i.v. and i.n. or a single dose of 0.25 mg/kg sgGFP following i.c.. knockout efficiency was assessed after seven days using Sanger Sequencing and Inference of CRISPR Edits (ICE) analysis. In vivo, the biodistribution of DiR labelled SNALPs (SNALPs-DiR) was assessed at 24h post-administration using IVIS Lumina Series III. Results: Serum-stable SNALPs produced were 130-140 nm in diameter with ~90% nucleic acid loading efficiency. SNALPs could reach and stay in the brain for up to 24h following i.v.; i.n. and i.c. administration. Decreasing GFP expression (around 50% after i.v. and i.c. and 20% following i.n.) was confirmed by optical imaging. Despite the small number of mice used, ICE analysis confirmed GFP knockout in mice brains. Additional studies are currently taking place to increase mice numbers. Conclusion: Results confirmed efficient gene knockout achieved by SNALPs in Rosa26-Cas9 knock-in mice expressing GFP following different routes of administrations in the following order i.v.= i.c.> i.n. Each of the administration routes has its pros and cons. The next stages of the project involve assessing gene-editing efficiency in wild-type mice and replacing GFP as a model target with therapeutic target genes implicated in Motor Neuron Disease pathology.

Keywords: CRISPR, nanoparticles, brain diseases, administration routes

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