Search results for: recalcitrant organic compounds
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 4357

Search results for: recalcitrant organic compounds

367 Xylanase Impact beyond Performance: A Prebiotic Approach in Laying Hens

Authors: Veerle Van Hoeck, Ingrid Somers, Dany Morisset

Abstract:

Anti-nutritional factors such as non-starch polysaccharides (NSP) are present in viscous cereals used to feed poultry. Therefore, exogenous carbohydrases are commonly added to monogastric feed to degrade these NSP. Our hypothesis is that xylanase not only improves laying hen performance and digestibility but also induces a significant shift in microbial composition within the intestinal tract and, thereby, can cause a prebiotic effect. In this context, a better understanding of whether and how the chicken gut flora can be modulated by xylanase is needed. To do so, in the herein laying hen study, the effects of dietary supplementation of xylanase on performance, digestibility, and cecal microbiome were evaluated. A total of 96 HiSex laying hens was used in this experiment (3 diets and 16 replicates of 2 hens). Xylanase was added to the diets at concentrations of 0, 45,000 (15 g/t XygestTM HT) and 90,000 U/kg (30 g/t Xygest HT). The diets were based on wheat (~55 %), soybean, and sunflower meal. The lowest dosage, 45,000 U/kg, significantly increased average egg weight and improved feed efficiency compared to the control treatment (p < 0.05). Egg quality parameters were significantly improved in the experiment in response to the xylanase addition. For example, during the last 28 days of the trial, the 45,000 U/kg and the 90,000 U/kg treatments exhibited an increase in Haugh units and albumin heights (p < 0.05). Compared with the control, organic matter digestibility and N retention were drastically improved in the 45,000 U/kg treatment group, which implies better nutrient digestibility at this lowest recommended dosage compared to the control (p < 0.05). Furthermore, gross energy and crude fat digestibility were improved significantly for birds fed 90,000 U/kg group compared to the control. Importantly, 16S rRNA gene analysis revealed that xylanase at 45,000 U/kg dosages can exert a prebiotic effect. This conclusion was drawn based on studying the sequence variation in the 16S rRNA gene in order to characterize diverse microbial communities of the cecal content. A significant increase in beneficial bacteria (Lactobacilli spp and Enterococcus casseliflavus) was documented when adding 45,000 U/kg xylanase to the diet of laying hens. In conclusion, dietary supplementation of xylanase, even at the lowest dose of (45,000 U/kg), significantly improved laying hen performance and digestibility. Furthermore, it is generally accepted that a proper bacterial balance between the number of beneficial bacteria and pathogenic bacteria in the intestine is vital for the host. It seems that the xylanase enzyme is able to modulate the laying hen microbiome beneficially and thus exerts a prebiotic effect. This microbiome plasticity in response to the xylanase provides an attractive target for stimulating intestinal health.

Keywords: laying hen, prebiotic, XygestTM HT, xylanase

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366 Medicinal Plants: An Antiviral Depository with Complex Mode of Action

Authors: Daniel Todorov, Anton Hinkov, Petya Angelova, Kalina Shishkova, Venelin Tsvetkov, Stoyan Shishkov

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Human herpes viruses (HHV) are ubiquitous pathogens with a pandemic spread across the globe. HHV type 1 is the main causative agent of cold sores and fever blisters around the mouth and on the face, whereas HHV type 2 is generally responsible for genital herpes outbreaks. The treatment of both viruses is more or less successful with antivirals from the nucleoside analogues group. Their wide application increasingly leads to the emergence of resistant mutants In the past, medicinal plants have been used to treat a number of infectious and non-infectious diseases. Their diversity and ability to produce the vast variety of secondary metabolites according to the characteristics of the environment give them the potential to help us in our warfare with viral infections. The variable chemical characteristics and complex composition is an advantage in the treatment of herpes since the emergence of resistant mutants is significantly complicated. The screening process is difficult due to the lack of standardization. That is why it is especially important to follow the mechanism of antiviral action of plants. On the one hand, it may be expected to interact with its compounds, resulting in enhanced antiviral effects, and the most appropriate environmental conditions can be chosen to maximize the amount of active secondary metabolites. During our study, we followed the activity of various plant extracts on the viral replication cycle as well as their effect on the extracellular virion. We obtained our results following the logical sequence of the experimental settings - determining the cytotoxicity of the extracts, evaluating the overall effect on viral replication and extracellular virion.During our research, we have screened a variety of plant extracts for their antiviral activity against both virus replication and the virion itself. We investigated the effect of the extracts on the individual stages of the viral replication cycle - viral adsorption, penetration and the effect on replication depending on the time of addition. If there are positive results in the later experiments, we had studied the activity over viral adsorption, penetration and the effect of replication according to the time of addition. Our results indicate that some of the extracts from the Lamium album have several targets. The first stages of the viral life cycle are most affected. Several of our active antiviral agents have shown an effect on extracellular virion and adsorption and penetration processes. Our research over the last decade has shown several curative antiviral plants - some of which are from the Lamiacea family. The rich set of active ingredients of the plants in this family makes them a good source of antiviral preparation.

Keywords: human herpes virus, antiviral activity, Lamium album, Nepeta nuda

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365 Development and Characterization of Novel Topical Formulation Containing Niacinamide

Authors: Sevdenur Onger, Ali Asram Sagiroglu

Abstract:

Hyperpigmentation is a cosmetically unappealing skin problem caused by an overabundance of melanin in the skin. Its pathophysiology is caused by melanocytes being exposed to paracrine melanogenic stimuli, which can upregulate melanogenesis-related enzymes (such as tyrosinase) and cause melanosome formation. Tyrosinase is linked to the development of melanosomes biochemically, and it is the main target of hyperpigmentation treatment. therefore, decreasing tyrosinase activity to reduce melanosomes has become the main target of hyperpigmentation treatment. Niacinamide (NA) is a natural chemical found in a variety of plants that is used as a skin-whitening ingredient in cosmetic formulations. NA decreases melanogenesis in the skin by inhibiting melanosome transfer from melanocytes to covering keratinocytes. Furthermore, NA protects the skin from reactive oxygen species and acts as a main barrier with the skin, reducing moisture loss by increasing ceramide and fatty acid synthesis. However, it is very difficult for hydrophilic compounds such as NA to penetrate deep into the skin. Furthermore, because of the nicotinic acid in NA, it is an irritant. As a result, we've concentrated on strategies to increase NA skin permeability while avoiding its irritating impacts. Since nanotechnology can affect drug penetration behavior by controlling the release and increasing the period of permanence on the skin, it can be a useful technique in the development of whitening formulations. Liposomes have become increasingly popular in the cosmetics industry in recent years due to benefits such as their lack of toxicity, high penetration ability in living skin layers, ability to increase skin moisture by forming a thin layer on the skin surface, and suitability for large-scale production. Therefore, liposomes containing NA were developed for this study. Different formulations were prepared by varying the amount of phospholipid and cholesterol and examined in terms of particle sizes, polydispersity index (PDI) and pH values. The pH values of the produced formulations were determined to be suitable with the pH value of the skin. Particle sizes were determined to be smaller than 250 nm and the particles were found to be of homogeneous size in the formulation (pdi<0.30). Despite the important advantages of liposomal systems, they have low viscosity and stability for topical use. For these reasons, in this study, liposomal cream formulations have been prepared for easy topical application of liposomal systems. As a result, liposomal cream formulations containing NA have been successfully prepared and characterized. Following the in-vitro release and ex-vivo diffusion studies to be conducted in the continuation of the study, it is planned to test the formulation that gives the most appropriate result on the volunteers after obtaining the approval of the ethics committee.

Keywords: delivery systems, hyperpigmentation, liposome, niacinamide

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364 Effects of the Natural Compound on SARS-CoV-2 Spike Protein-Mediated Metabolic Alteration in THP-1 Cells Explored by the ¹H-NMR-Based Metabolomics Approach

Authors: Gyaltsen Dakpa, K. J. Senthil Kumar, Nai-Wen Tsao, Sheng-Yang Wang

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Context: Coronavirus disease 2019 (COVID-19) is a severe respiratory illness caused by the SARS-CoV-2 virus. One of the hallmarks of COVID-19 is a change in metabolism, which can lead to increased severity and mortality. The mechanism of SARS-CoV-2-mediated perturbations of metabolic pathways has yet to be fully understood. Research Aim: This study aimed to investigate the metabolic alteration caused by SARS-CoV-2 spike protein in Phorbol 12-myristate 13-acetate (PMA)-induced human monocytes (THP-1) and to examine the regulatory effect of natural compounds like Antcins A on SARS-CoV-2 spike protein-induced metabolic alteration. Methodology: The study used a combination of proton nuclear magnetic resonance (1H-NMR) and MetaboAnalyst 5.0 software. THP-1 cells were treated with SARS-CoV-2 spike protein or control, and the metabolomic profiles of the cells were compared. Antcin A was also added to the cells to assess its regulatory effect on SARS-CoV-2 spike protein-induced metabolic alteration. Findings: The study results showed that treatment with SARS-CoV-2 spike protein significantly altered the metabolomic profiles of THP-1 cells. Eight metabolites, including glycerol-phosphocholine, glycine, canadine, sarcosine, phosphoenolpyruvic acid, glutamine, glutamate, and N, N-dimethylglycine, were significantly different between control and spike-protein treatment groups. Antcin A significantly reversed the changes in these metabolites. In addition, treatment with antacid A significantly inhibited SARS-CoV-2 spike protein-mediated up-regulation of TLR-4 and ACE2 receptors. Theoretical Importance The findings of this study suggest that SARS-CoV-2 spike protein can cause significant metabolic alterations in THP-1 cells. Antcin A, a natural compound, has the potential to reverse these metabolic alterations and may be a potential candidate for developing preventive or therapeutic agents for COVID-19. Data Collection: The data for this study was collected from THP-1 cells that were treated with SARS-CoV-2 spike protein or a control. The metabolomic profiles of the cells were then compared using 1H-NMR and MetaboAnalyst 5.0 software. Analysis Procedures: The metabolomic profiles of the THP-1 cells were analyzed using 1H-NMR and MetaboAnalyst 5.0 software. The software was used to identify and quantify the cells' metabolites and compare the control and spike-protein treatment groups. Questions Addressed: The question addressed by this study was whether SARS-CoV-2 spike protein could cause metabolic alterations in THP-1 cells and whether Antcin A can reverse these alterations. Conclusion: The findings of this study suggest that SARS-CoV-2 spike protein can cause significant metabolic alterations in THP-1 cells. Antcin A, a natural compound, has the potential to reverse these metabolic alterations and may be a potential candidate for developing preventive or therapeutic agents for COVID-19.

Keywords: SARS-CoV-2-spike, ¹H-NMR, metabolomics, antcin-A, taiwanofungus camphoratus

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363 The Role of EDTA and EDDS in Reducing Metal Toxicity for Aquaculture Shellfish Perna canaliculus

Authors: Daniel R. McDougall, Martin D. de Jonge, Gordon M. Miskelly, Duncan J. McGillivray, Andrew G. Jeffs

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The chelating agent ethylenediaminetetraacetic acid (EDTA) is commonly added as a cure-all to seawater in aquaculture hatcheries around the world to reduce heavy metal toxicity, significantly improve the survival of larval shellfish, and to therefore improve the overall production efficiency of the aquaculture industry. However, EDTA is not a biodegradable chemical and is considered to be a persistent organic pollutant, which will accumulate in the environment over time. This makes the use of EDTA unsustainable environmentally, and therefore alternatives should be considered. Ethylenediaminedisuccinic acid (EDDS) is a biodegradable alternative to EDTA with very similar metal chelation properties. This study investigates the effect of EDTA and EDDS at two different concentrations, on metal concentrations found within developing New Zealand green-lipped mussel (Perna canaliculus) larvae. P. canaliculus is New Zealand’s main shellfish aquaculture species, providing a major export for New Zealand’s economy, with excellent potential for increased production in the near future. It is well known that the early stages of bivalve development are the most vulnerable to metal toxicity and P. canaliculus is no exception. The commercially used concentration (12 µmol L⁻¹) of EDTA added to P. canaliculus larval rearing tanks often increases the yield of D-larvae by over 80%. This concentration of EDTA and EDDS will be tested in this study, along with a lower concentration (3 µmol L⁻¹). After 48 hours of larval development, the D-larvae will be analyzed for heavy metal content with Inductively Coupled Plasma Mass Spectrometry (ICP-MS) and heavy metal distribution with synchrotron X-ray Fluorescence Microscopy (XFM). In this study, we found that EDDS also improves the yield of P. canaliculus larvae and could be a viable alternative to EDTA in aquaculture. Furthermore, results suggest a higher concentration of chelating agent is more effective for improving the yield of developing P. canaliculus larvae. Metals with significant differences in concentration with the addition of EDTA were Cr, Cu, Zn, Cd and Pb (P < 0.05). We observed for the first time to the author’s best knowledge, metal distribution within 100 µm P. canaliculus D-larvae using synchrotron XFM and found changes in the distribution of metals with the addition of EDTA. XFM also has the potential to provide information about the chemical state of the metals within mussel larvae. This research provides greater insight into the reasons for the effectiveness of adding the chelating agent to aquaculture culture water, and a more environmentally conscious alternative to the currently used EDTA, which could be extremely valuable for the aquaculture industry.

Keywords: EDDS, EDTA, heavy metals, P. canaliculus, toxicity, water treatment

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362 Linking Soil Spectral Behavior and Moisture Content for Soil Moisture Content Retrieval at Field Scale

Authors: Yonwaba Atyosi, Moses Cho, Abel Ramoelo, Nobuhle Majozi, Cecilia Masemola, Yoliswa Mkhize

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Spectroscopy has been widely used to understand the hyperspectral remote sensing of soils. Accurate and efficient measurement of soil moisture is essential for precision agriculture. The aim of this study was to understand the spectral behavior of soil at different soil water content levels and identify the significant spectral bands for soil moisture content retrieval at field-scale. The study consisted of 60 soil samples from a maize farm, divided into four different treatments representing different moisture levels. Spectral signatures were measured for each sample in laboratory under artificial light using an Analytical Spectral Device (ASD) spectrometer, covering a wavelength range from 350 nm to 2500 nm, with a spectral resolution of 1 nm. The results showed that the absorption features at 1450 nm, 1900 nm, and 2200 nm were particularly sensitive to soil moisture content and exhibited strong correlations with the water content levels. Continuum removal was developed in the R programming language to enhance the absorption features of soil moisture and to precisely understand its spectral behavior at different water content levels. Statistical analysis using partial least squares regression (PLSR) models were performed to quantify the correlation between the spectral bands and soil moisture content. This study provides insights into the spectral behavior of soil at different water content levels and identifies the significant spectral bands for soil moisture content retrieval. The findings highlight the potential of spectroscopy for non-destructive and rapid soil moisture measurement, which can be applied to various fields such as precision agriculture, hydrology, and environmental monitoring. However, it is important to note that the spectral behavior of soil can be influenced by various factors such as soil type, texture, and organic matter content, and caution should be taken when applying the results to other soil systems. The results of this study showed a good agreement between measured and predicted values of Soil Moisture Content with high R2 and low root mean square error (RMSE) values. Model validation using independent data was satisfactory for all the studied soil samples. The results has significant implications for developing high-resolution and precise field-scale soil moisture retrieval models. These models can be used to understand the spatial and temporal variation of soil moisture content in agricultural fields, which is essential for managing irrigation and optimizing crop yield.

Keywords: soil moisture content retrieval, precision agriculture, continuum removal, remote sensing, machine learning, spectroscopy

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361 Conservation Agriculture under Mediterranean Climate: Effects on below and Above-Ground Processes during Wheat Cultivation

Authors: Vasiliki Kolake, Christos Kavalaris, Sofia Megoudi, Maria Maxouri, Panagiotis A. Karas, Aris Kyparissis, Efi Levizou

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Conservation agriculture (CA), is a production system approach that can tackle the challenges of climate change mainly through facilitating carbon storage into the soil and increasing crop resilience. This is extremely important for the vulnerable Mediterranean agroecosystems, which already face adverse environmental conditions. The agronomic practices used in CA, i.e. permanent soil cover and no-tillage, result in reduced soil erosion and increased soil organic matter, preservation of water and improvement of quality and fertility of the soil in the long-term. Thus the functional characteristics and processes of the soil are considerably affected by the implementation of CA. The aim of the present work was to assess the effects of CA on soil nitrification potential and mycorrhizal colonization about the above-ground production in a wheat field. Two adjacent but independent field sites of 1.5ha each were used (Thessaly plain, Central Greece), comprising the no-till and conventional tillage treatments. The no-tillage site was covered by residues of the previous crop (cotton). Potential nitrification and the nitrate and ammonium content of the soil were measured at two different soil depths (3 and 15cm) at 20-days intervals throughout the growth period. Additionally, the leaf area index (LAI) was monitored at the same time-course. The mycorrhizal colonization was measured at the commencement and end of the experiment. At the final harvest, total yield and plant biomass were also recorded. The results indicate that wheat yield was considerably favored by CA practices, exhibiting a 42% increase compared to the conventional tillage treatment. The superior performance of the CA crop was also depicted in the above-ground plant biomass, where a 26% increase was recorded. LAI, which is considered a reliable growth index, did not show statistically significant differences between treatments throughout the growth period. On the contrary, significant differences were recorded in endomycorrhizal colonization one day before the final harvest, with CA plants exhibiting 20% colonization, while the conventional tillage plants hardly reached 1%. The on-going analyses of potential nitrification measurements, as well as nitrate and ammonium determination, will shed light on the effects of CA on key processes in the soil. These results will integrate the assessment of CA impact on certain below and above-ground processes during wheat cultivation under the Mediterranean climate.

Keywords: conservation agriculture, LAI, mycorrhizal colonization, potential nitrification, wheat, yield

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360 Comparison of the Toxicity of Silver and Gold Nanoparticles in Murine Fibroblasts

Authors: Šárka Hradilová, Aleš Panáček, Radek Zbořil

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Nanotechnologies are considered the most promising fields with high added value, brings new possibilities in various sectors from industry to medicine. With the growing of interest in nanomaterials and their applications, increasing nanoparticle production leads to increased exposure of people and environment with ‘human made’ nanoparticles. Nanoparticles (NPs) are clusters of atoms in the size range of 1–100 nm. Metal nanoparticles represent one of the most important and frequently used types of NPs due to their unique physical, chemical and biological properties, which significantly differ from those of bulk material. Biological properties including toxicity of metal nanoparticles are generally determined by their size, size distribution, shape, surface area, surface charge, surface chemistry, stability in the environment and ability to release metal ions. Therefore, the biological behavior of NPs and their possible adverse effect cannot be derived from the bulk form of material because nanoparticles show unique properties and interactions with biological systems just due to their nanodimensions. Silver and gold NPs are intensively studied and used. Both can be used for instance in surface enhanced Raman spectroscopy, a considerable number of applications of silver NPs is associated with antibacterial effects, while gold NPs are associated with cancer treatment and bio imaging. Antibacterial effects of silver ions are known for centuries. Silver ions and silver-based compounds are highly toxic to microorganisms. Toxic properties of silver NPs are intensively studied, but the mechanism of cytoxicity is not fully understood. While silver NPs are considered toxic, gold NPs are referred to as toxic but also innocuous for eukaryotic cells. Therefore, gold NPs are used in various biological applications without a risk of cell damaging, even when we want to suppress the growth of cancer cells. Thus, gold NPs are toxic or harmless. Because most studies comparing particles of various sizes prepared in various ways, and testing is performed on different cell lines, it is very difficult to generalize. The novelty and significance of our research is focused to the complex biological effects of silver and gold NPs prepared by the same method, have the same parameters and the same stabilizer. That is why we can compare the biological effects of pure nanometals themselves based on their chemical nature without the influence of other variable. Aim of our study therefore is to compare the cytotoxic effect of two types of noble metal NPs focusing on the mechanisms that contribute to cytotoxicity. The study was conducted on murine fibroblasts by selected common used tests. Each of these tests monitors the selected area related to toxicity and together provides a comprehensive view on the issue of interactions of nanoparticles and living cells.

Keywords: cytotoxicity, gold nanoparticles, mechanism of cytotoxicity, silver nanoparticles

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359 Enhancing Air Quality: Investigating Filter Lifespan and Byproducts in Air Purification Solutions

Authors: Freja Rydahl Rasmussen, Naja Villadsen, Stig Koust

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Air purifiers have become widely implemented in a wide range of settings, including households, schools, institutions, and hospitals, as they tackle the pressing issue of indoor air pollution. With their ability to enhance indoor air quality and create healthier environments, air purifiers are particularly vital when ventilation options are limited. These devices incorporate a diverse array of technologies, including HEPA filters, active carbon filters, UV-C light, photocatalytic oxidation, and ionizers, each designed to combat specific pollutants and improve air quality within enclosed spaces. However, the safety of air purifiers has not been investigated thoroughly, and many questions still arise when applying them. Certain air purification technologies, such as UV-C light or ionization, can unintentionally generate undesirable byproducts that can negatively affect indoor air quality and health. It is well-established that these technologies can inadvertently generate nanoparticles or convert common gaseous compounds into harmful ones, thus exacerbating air pollution. However, the formation of byproducts can vary across products, necessitating further investigation. There is a particular concern about the formation of the carcinogenic substance formaldehyde from common gases like acetone. Many air purifiers use mechanical filtration to remove particles, dust, and pollen from the air. Filters need to be replaced periodically for optimal efficiency, resulting in an additional cost for end-users. Currently, there are no guidelines for filter lifespan, and replacement recommendations solely rely on manufacturers. A market screening revealed that manufacturers' recommended lifespans vary greatly (from 1 month to 10 years), and there is a need for general recommendations to guide consumers. Activated carbon filters are used to adsorb various types of chemicals that can pose health risks or cause unwanted odors. These filters have a certain capacity before becoming saturated. If not replaced in a timely manner, the adsorbed substances are likely to be released from the filter through off-gassing or losing adsorption efficiency. The goal of this study is to investigate the lifespan of filters as well as investigate the potentially harmful effects of air purifiers. Understanding the lifespan of filters used in air purifiers and the potential formation of harmful byproducts is essential for ensuring their optimal performance, guiding consumers in their purchasing decisions, and establishing industry standards for safer and more effective air purification solutions. At this time, a selection of air purifiers has been chosen, and test methods have been established. In the following 3 months, the tests will be conducted, and the results will be ready for presentation later.

Keywords: air purifiers, activated carbon filters, byproducts, clean air, indoor air quality

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358 Agrowastes to Edible Hydrogels through Bio Nanotechnology Interventions: Bioactive from Mandarin Peels

Authors: Niharika Kaushal, Minni Singh

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Citrus fruits contain an abundance of phytochemicals that can promote health. A substantial amount of agrowaste is produced from the juice processing industries, primarily peels and seeds. This leftover agrowaste is a reservoir of nutraceuticals, particularly bioflavonoids which render it antioxidant and potentially anticancerous. It is, therefore, favorable to utilize this biomass and contribute towards sustainability in a manner that value-added products may be derived from them, nutraceuticals, in this study. However, the pre-systemic metabolism of flavonoids in the gastric phase limits the effectiveness of these bioflavonoids derived from mandarin biomass. In this study, ‘kinnow’ mandarin (Citrus nobilis X Citrus deliciosa) biomass was explored for its flavonoid profile. This work entails supercritical fluid extraction and identification of bioflavonoids from mandarin biomass. Furthermore, to overcome the limitations of these flavonoids in the gastrointestinal tract, a double-layered vehicular mechanism comprising the fabrication of nanoconjugates and edible hydrogels was adopted. Total flavonoids in the mandarin peel extract were estimated by the aluminum chloride complexation method and were found to be 47.3±1.06 mg/ml rutin equivalents as total flavonoids. Mass spectral analysis revealed the abundance of polymethoxyflavones (PMFs), nobiletin and tangeretin as the major flavonoids in the extract, followed by hesperetin and naringenin. Furthermore, the antioxidant potential was analyzed by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method, which showed an IC50 of 0.55μg/ml. Nanoconjugates were fabricated via the solvent evaporation method, which was further impregnated into hydrogels. Additionally, the release characteristics of nanoconjugate-laden hydrogels in a simulated gastrointestinal environment were studied. The PLGA-PMFs nanoconjugates exhibited a particle size between 200-250nm having a smooth and spherical shape as revealed by FE-SEM. The impregnated alginate hydrogels offered a dense network that ensured the holding of PLGA-PMF nanoconjugates, as confirmed by Cryo-SEM images. Rheological studies revealed the shear-thinning behavior of hydrogels and their high resistance to deformation. Gastrointestinal studies showed a negligible 4.0% release of flavonoids in the gastric phase, followed by a sustained release over the next hours in the intestinal environment. Therefore, based on the enormous potential of recovering nutraceuticals from agro-processing wastes, further augmented by nanotechnological interventions for enhancing the bioefficacy of these compounds, lays the foundation for exploring the path towards the development of value-added products, thereby contributing towards the sustainable use of agrowaste.

Keywords: agrowaste, gastrointestinal, hydrogel, nutraceuticals

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357 Design, Simulation and Fabrication of Electro-Magnetic Pulse Welding Coil and Initial Experimentation

Authors: Bharatkumar Doshi

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Electro-Magnetic Pulse Welding (EMPW) is a solid state welding process carried out at almost room temperature, in which joining is enabled by high impact velocity deformation. In this process, high voltage capacitor’s stored energy is discharged in an EM coil resulting in a damped, sinusoidal current with an amplitude of several hundred kiloamperes. Due to these transient magnetic fields of few tens of Tesla near the coil is generated. As the conductive (tube) part is positioned in this area, an opposing eddy current is induced in this part. Consequently, high Lorentz forces act on the part, leading to acceleration away from the coil. In case of a tube, it gets compressed under forming velocities of more than 300 meters per second. After passing the joining gap it collides with the second metallic joining rod, leading to the formation of a jet under appropriate collision conditions. Due to the prevailing high pressure, metallurgical bonding takes place. A characteristic feature is the wavy interface resulting from the heavy plastic deformations. In the process, the formation of intermetallic compounds which might deteriorate the weld strength can be avoided, even for metals with dissimilar thermal properties. In order to optimize the process parameters like current, voltage, inductance, coil dimensions, workpiece dimensions, air gap, impact velocity, effective plastic strain, shear stress acting in the welding zone/impact zone etc. are very critical and important to establish. These process parameters could be determined by simulation using Finite Element Methods (FEM) in which electromagnetic –structural couple field analysis is performed. The feasibility of welding could thus be investigated by varying the parameters in the simulation using COMSOL. Simulation results shall be applied in performing the preliminary experiments of welding the different alloy steel tubes and/or alloy steel to other materials. The single turn coil (S.S.304) with field shaper (copper) has been designed and manufactured. The preliminary experiments are performed using existing EMPW facility available Institute for Plasma Research, Gandhinagar, India. The experiments are performed at 22kV charged into 64µF capacitor bank and the energy is discharged into single turn EM coil. Welding of axi-symetric components such as aluminum tube and rod has been proven experimentally using EMPW techniques. In this paper EM coil design, manufacturing, Electromagnetic-structural FEM simulation of Magnetic Pulse Welding and preliminary experiment results is reported.

Keywords: COMSOL, EMPW, FEM, Lorentz force

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356 Risk Assessment of Lead Element in Red Peppers Collected from Marketplaces in Antalya, Southern Turkey

Authors: Serpil Kilic, Ihsan Burak Cam, Murat Kilic, Timur Tongur

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Interest in the lead (Pb) has considerably increased due to knowledge about the potential toxic effects of this element, recently. Exposure to heavy metals above the acceptable limit affects human health. Indeed, Pb is accumulated through food chains up to toxic concentrations; therefore, it can pose an adverse potential threat to human health. A sensitive and reliable method for determination of Pb element in red pepper were improved in the present study. Samples (33 red pepper products having different brands) were purchased from different markets in Turkey. The selected method validation criteria (linearity, Limit of Detection, Limit of Quantification, recovery, and trueness) demonstrated. Recovery values close to 100% showed adequate precision and accuracy for analysis. According to the results of red pepper analysis, all of the tested lead element in the samples was determined at various concentrations. A Perkin- Elmer ELAN DRC-e model ICP-MS system was used for detection of Pb. Organic red pepper was used to obtain a matrix for all method validation studies. The certified reference material, Fapas chili powder, was digested and analyzed, together with the different sample batches. Three replicates from each sample were digested and analyzed. The results of the exposure levels of the elements were discussed considering the scientific opinions of the European Food Safety Authority (EFSA), which is the European Union’s (EU) risk assessment source associated with food safety. The Target Hazard Quotient (THQ) was described by the United States Environmental Protection Agency (USEPA) for the calculation of potential health risks associated with long-term exposure to chemical pollutants. THQ value contains intake of elements, exposure frequency and duration, body weight and the oral reference dose (RfD). If the THQ value is lower than one, it means that the exposed population is assumed to be safe and 1 < THQ < 5 means that the exposed population is in a level of concern interval. In this study, the THQ of Pb was obtained as < 1. The results of THQ calculations showed that the values were below one for all the tested, meaning the samples did not pose a health risk to the local population. This work was supported by The Scientific Research Projects Coordination Unit of Akdeniz University. Project Number: FBA-2017-2494.

Keywords: lead analyses, red pepper, risk assessment, daily exposure

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355 A Comprehensive Study on Freshwater Aquatic Life Health Quality Assessment Using Physicochemical Parameters and Planktons as Bio Indicator in a Selected Region of Mahaweli River in Kandy District, Sri Lanka

Authors: S. M. D. Y. S. A. Wijayarathna, A. C. A. Jayasundera

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Mahaweli River is the longest and largest river in Sri Lanka and it is the major drinking water source for a large portion of 2.5 million inhabitants in the Central Province. The aim of this study was to the determination of water quality and aquatic life health quality in a selected region of Mahaweli River. Six sampling locations (Site 1: 7° 16' 50" N, 80° 40' 00" E; Site 2: 7° 16' 34" N, 80° 40' 27" E; Site 3: 7° 16' 15" N, 80° 41' 28" E; Site 4: 7° 14' 06" N, 80° 44' 36" E; Site 5: 7° 14' 18" N, 80° 44' 39" E; Site 6: 7° 13' 32" N, 80° 46' 11" E) with various anthropogenic activities at bank of the river were selected for a period of three months from Tennekumbura Bridge to Victoria Reservoir. Temperature, pH, Electrical Conductivity (EC), Total Dissolved Solids (TDS), Dissolved Oxygen (DO), 5-day Biological Oxygen Demand (BOD5), Total Suspended Solids (TSS), hardness, the concentration of anions, and metal concentration were measured according to the standard methods, as physicochemical parameters. Planktons were considered as biological parameters. Using a plankton net (20 µm mesh size), surface water samples were collected into acid washed dried vials and were stored in an ice box during transportation. Diversity and abundance of planktons were identified within 4 days of sample collection using standard manuals of plankton identification under the light microscope. Almost all the measured physicochemical parameters were within the CEA standards limits for aquatic life, Sri Lanka Standards (SLS) or World Health Organization’s Guideline for drinking water. Concentration of orthophosphate ranged between 0.232 to 0.708 mg L-1, and it has exceeded the standard limit of aquatic life according to CEA guidelines (0.400 mg L-1) at Site 1 and Site 2, where there is high disturbance by cultivations and close households. According to the Pearson correlation (significant correlation at p < 0.05), it is obvious that some physicochemical parameters (temperature, DO, TDS, TSS, phosphate, sulphate, chloride fluoride, and sodium) were significantly correlated to the distribution of some plankton species such as Aulocoseira, Navicula, Synedra, Pediastrum, Fragilaria, Selenastrum, Oscillataria, Tribonema and Microcystis. Furthermore, species that appear in blooms (Aulocoseira), organic pollutants (Navicula), and phosphate high eutrophic water (Microcystis) were found, indicating deteriorated water quality in Mahaweli River due to agricultural activities, solid waste disposal, and release of domestic effluents. Therefore, it is necessary to improve environmental monitoring and management to control the further deterioration of water quality of the river.

Keywords: bio indicator, environmental variables, planktons, physicochemical parameters, water quality

Procedia PDF Downloads 109
354 Effect of Fermented Orange Juice Intake on Urinary 6‑Sulfatoxymelatonin in Healthy Volunteers

Authors: I. Cerrillo, A. Carrillo-Vico, M. A. Ortega, B. Escudero-López, N. Álvarez-Sánchez, F. Martín, M. S. Fernández-Pachón

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Melatonin is a bioactive compound involved in multiple biological activities such as glucose tolerance, circadian rhythm regulation, antioxidant defense or immune system action. In elderly subjects the intake of foods and drinks rich in melatonin is very important due to its endogenous level decreases with age. Alcoholic fermentation is a process carried out in fruits, vegetables and legumes to obtain new products with improved bioactive compounds profile in relation to original substrates. Alcoholic fermentation process carried out by Saccharomycetaceae var. Pichia kluyveri induces an important synthesis of melatonin in orange juice. A novel beverage derived of fermented orange juice could be a promising source of this bioactive compound. The aim of the present study was to determine whether the acute intake of fermented orange juice increase the levels of urinary 6-sulfatoxymelatonin in healthy humans. Nine healthy volunteers (7 women and 2 men), aged between 20 and 25 years old and BMI of 21.1  2.4 kg/m2, were recruited. On the study day, participants ingested 500 mL of fermented orange juice. The first urine collection was made before fermented orange juice consumption (basal). The rest of urine collections were made in the following time intervals after fermented orange juice consumption: 0-2, 2-5, 5-10, 10- 15 and 15-24 hours. During the experimental period only the consumption of water was allowed. At lunch time a meal was provided (60 g of white bread, two slices of ham, a slice of cheese, 125 g of sweetened natural yoghurt and water). The subjects repeated the protocol with orange juice following a 2-wk washout period between both types of beverages. The levels of 6-sulfatoxymelatonin (6-SMT) were measured in urine recollected at different time points using the Melatonin-Sulfate Urine ELISA (IBL International GMBH, Hamburg, Germany). Levels of 6-SMT were corrected to those of creatinine for each sample. A significant (p < 0.05) increase in urinary 6-SMT levels was observed between 2-5 hours after fermented orange juice ingestion with respect to basal values (increase of 67,8 %). The consumption of orange juice did not induce any significant change in urinary 6-SMT levels. In addition, urinary 6-SMT levels obtained between 2-5 hours after fermented orange juice ingestion (115,6 ng/mg) were significantly different (p < 0.05) from those of orange juice (42,4 ng/mg). The enhancement of urinary 6-SMT after the ingestion of 500 mL of fermented orange juice in healthy humans compared to orange juice could be an important advantage of this novel product as an excellent source of melatonin. Fermented orange juice could be a new functional food, and its consumption could exert a potentially positive effect on health in both the maintenance of health status and the prevention of chronic diseases.

Keywords: fermented orange juice, functional beverage, healthy human, melatonin

Procedia PDF Downloads 411
353 Control of Doxorubicin Release Rate from Magnetic PLGA Nanoparticles Using a Non-Permanent Magnetic Field

Authors: Inês N. Peça , A. Bicho, Rui Gardner, M. Margarida Cardoso

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Inorganic/organic nanocomplexes offer tremendous scope for future biomedical applications, including imaging, disease diagnosis and drug delivery. The combination of Fe3O4 with biocompatible polymers to produce smart drug delivery systems for use in pharmaceutical formulation present a powerful tool to target anti-cancer drugs to specific tumor sites through the application of an external magnetic field. In the present study, we focused on the evaluation of the effect of the magnetic field application time on the rate of drug release from iron oxide polymeric nanoparticles. Doxorubicin, an anticancer drug, was selected as the model drug loaded into the nanoparticles. Nanoparticles composed of poly(d-lactide-co-glycolide (PLGA), a biocompatible polymer already approved by FDA, containing iron oxide nanoparticles (MNP) for magnetic targeting and doxorubicin (DOX) were synthesized by the o/w solvent extraction/evaporation method and characterized by scanning electron microscopy (SEM), by dynamic light scattering (DLS), by inductively coupled plasma-atomic emission spectrometry and by Fourier transformed infrared spectroscopy. The produced particles yielded smooth surfaces and spherical shapes exhibiting a size between 400 and 600 nm. The effect of the magnetic doxorubicin loaded PLGA nanoparticles produced on cell viability was investigated in mammalian CHO cell cultures. The results showed that unloaded magnetic PLGA nanoparticles were nontoxic while the magnetic particles without polymeric coating show a high level of toxicity. Concerning the therapeutic activity doxorubicin loaded magnetic particles cause a remarkable enhancement of the cell inhibition rates compared to their non-magnetic counterpart. In vitro drug release studies performed under a non-permanent magnetic field show that the application time and the on/off cycle duration have a great influence with respect to the final amount and to the rate of drug release. In order to determine the mechanism of drug release, the data obtained from the release curves were fitted to the semi-empirical equation of the the Korsmeyer-Peppas model that may be used to describe the Fickian and non-Fickian release behaviour. Doxorubicin release mechanism has shown to be governed mainly by Fickian diffusion. The results obtained show that the rate of drug release from the produced magnetic nanoparticles can be modulated through the magnetic field time application.

Keywords: drug delivery, magnetic nanoparticles, PLGA nanoparticles, controlled release rate

Procedia PDF Downloads 263
352 Measurement of in-situ Horizontal Root Tensile Strength of Herbaceous Vegetation for Improved Evaluation of Slope Stability in the Alps

Authors: Michael T. Lobmann, Camilla Wellstein, Stefan Zerbe

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Vegetation plays an important role for the stabilization of slopes against erosion processes, such as shallow erosion and landslides. Plant roots reinforce the soil, increase soil cohesion and often cross possible shear planes. Hence, plant roots reduce the risk of slope failure. Generally, shrub and tree roots penetrate deeper into the soil vertically, while roots of forbs and grasses are concentrated horizontally in the topsoil and organic layer. Therefore, shrubs and trees have a higher potential for stabilization of slopes with deep soil layers than forbs and grasses. Consequently, research mainly focused on the vertical root effects of shrubs and trees. Nevertheless, a better understanding of the stabilizing effects of grasses and forbs is needed for better evaluation of the stability of natural and artificial slopes with herbaceous vegetation. Despite the importance of vertical root effects, field observations indicate that horizontal root effects also play an important role for slope stabilization. Not only forbs and grasses, but also some shrubs and trees form tight horizontal networks of fine and coarse roots and rhizomes in the topsoil. These root networks increase soil cohesion and horizontal tensile strength. Available methods for physical measurements, such as shear-box tests, pullout tests and singular root tensile strength measurement can only provide a detailed picture of vertical effects of roots on slope stabilization. However, the assessment of horizontal root effects is largely limited to computer modeling. Here, a method for measurement of in-situ cumulative horizontal root tensile strength is presented. A traction machine was developed that allows fixation of rectangular grass sods (max. 30x60cm) on the short ends with a 30x30cm measurement zone in the middle. On two alpine grass slopes in South Tyrol (northern Italy), 30x60cm grass sods were cut out (max. depth 20cm). Grass sods were pulled apart measuring the horizontal tensile strength over 30cm width over the time. The horizontal tensile strength of the sods was measured and compared for different soil depths, hydrological conditions, and root physiological properties. The results improve our understanding of horizontal root effects on slope stabilization and can be used for improved evaluation of grass slope stability.

Keywords: grassland, horizontal root effect, landslide, mountain, pasture, shallow erosion

Procedia PDF Downloads 171
351 Co-pyrolysis of Sludge and Kaolin/Zeolite to Stabilize Heavy Metals

Authors: Qian Li, Zhaoping Zhong

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Sewage sludge, a typical solid waste, has inevitably been produced in enormous quantities in China. Still worse, the amount of sewage sludge produced has been increasing due to rapid economic development and urbanization. Compared to the conventional method to treat sewage sludge, pyrolysis has been considered an economic and ecological technology because it can significantly reduce the sludge volume, completely kill pathogens, and produce valuable solid, gas, and liquid products. However, the large-scale utilization of sludge biochar has been limited due to the considerable risk posed by heavy metals in the sludge. Heavy metals enriched in pyrolytic biochar could be divided into exchangeable, reducible, oxidizable, and residual forms. The residual form of heavy metals is the most stable and cannot be used by organisms. Kaolin and zeolite are environmentally friendly inorganic minerals with a high surface area and heat resistance characteristics. So, they exhibit the enormous potential to immobilize heavy metals. In order to reduce the risk of leaching heavy metals in the pyrolysis biochar, this study pyrolyzed sewage sludge mixed with kaolin/zeolite in a small rotary kiln. The influences of additives and pyrolysis temperature on the leaching concentration and morphological transformation of heavy metals in pyrolysis biochar were investigated. The potential mechanism of stabilizing heavy metals in the co-pyrolysis of sludge blended with kaolin/zeolite was explained by scanning electron microscopy, X-ray diffraction, and specific surface area and porosity analysis. The European Community Bureau of Reference sequential extraction procedure has been applied to analyze the forms of heavy metals in sludge and pyrolysis biochar. All the concentrations of heavy metals were examined by flame atomic absorption spectrophotometry. Compared with the proportions of heavy metals associated with the F4 fraction in pyrolytic carbon prepared without additional agents, those in carbon obtained by co-pyrolysis of sludge and kaolin/zeolite increased. Increasing the additive dosage could improve the proportions of the stable fraction of various heavy metals in biochar. Kaolin exhibited a better effect on stabilizing heavy metals than zeolite. Aluminosilicate additives with excellent adsorption performance could capture more released heavy metals during sludge pyrolysis. Then heavy metal ions would react with the oxygen ions of additives to form silicate and aluminate, causing the conversion of heavy metals from unstable fractions (sulfate, chloride, etc.) to stable fractions (silicate, aluminate, etc.). This study reveals that the efficiency of stabilizing heavy metals depends on the formation of stable mineral compounds containing heavy metals in pyrolysis biochar.

Keywords: co-pyrolysis, heavy metals, immobilization mechanism, sewage sludge

Procedia PDF Downloads 70
350 Biopolymers: A Solution for Replacing Polyethylene in Food Packaging

Authors: Sonia Amariei, Ionut Avramia, Florin Ursachi, Ancuta Chetrariu, Ancuta Petraru

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The food industry is one of the major generators of plastic waste derived from conventional synthetic petroleum-based polymers, which are non-biodegradable, used especially for packaging. These packaging materials, after the food is consumed, accumulate serious environmental concerns due to the materials but also to the organic residues that adhere to them. It is the concern of specialists, researchers to eliminate problems related to conventional materials that are not biodegradable or unnecessary plastic and replace them with biodegradable and edible materials, supporting the common effort to protect the environment. Even though environmental and health concerns will cause more consumers to switch to a plant-based diet, most people will continue to add more meat to their diet. The paper presents the possibility of replacing the polyethylene packaging from the surface of the trays for meat preparations with biodegradable packaging obtained from biopolymers. During the storage of meat products may occur deterioration by lipids oxidation and microbial spoilage, as well as the modification of the organoleptic characteristics. For this reason, different compositions of polymer mixtures and film conditions for obtaining must be studied to choose the best packaging material to achieve food safety. The compositions proposed for packaging are obtained from alginate, agar, starch, and glycerol as plasticizers. The tensile strength, elasticity, modulus of elasticity, thickness, density, microscopic images of the samples, roughness, opacity, humidity, water activity, the amount of water transferred as well as the speed of water transfer through these packaging materials were analyzed. A number of 28 samples with various compositions were analyzed, and the results showed that the sample with the highest values for hardness, density, and opacity, as well as the smallest water vapor permeability, of 1.2903E-4 ± 4.79E-6, has the ratio of components as alginate: agar: glycerol (3:1.25:0.75). The water activity of the analyzed films varied between 0.2886 and 0.3428 (aw< 0.6), demonstrating that all the compositions ensure the preservation of the products in the absence of microorganisms. All the determined parameters allow the appreciation of the quality of the packaging films in terms of mechanical resistance, its protection against the influence of light, the transfer of water through the packaging. Acknowledgments: This work was supported by a grant of the Ministry of Research, Innovation, and Digitization, CNCS/CCCDI – UEFISCDI, project number PN-III-P2-2.1-PED-2019-3863, within PNCDI III.

Keywords: meat products, alginate, agar, starch, glycerol

Procedia PDF Downloads 178
349 The Effect of the Structural Arrangement of Binary Bisamide Organogelators on their Self-Assembly Behavior

Authors: Elmira Ghanbari, Jan Van Esch, Stephen J. Picken, Sahil Aggarwal

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Low-molecular-weight organogelators form gels by self-assembly into the crystalline network which immobilizes the organic solvent. For single bisamide organogelator systems, the effect of the molecular structure on the molecular interaction and their self-assembly behavior has been explored. The spatial arrangement of bisamide molecules in the gel-state is driven by a combination of hydrogen bonding and Van der Waals interactions. The hydrogen-bonding pattern between the amide groups of bisamide molecules is regulated by the number of methylene spacers; the even number of methylene spacers between two amide groups, in even-spaced bisamides, leads to the antiparallel position of amide groups within a molecule. An even-spaced bisamide molecule with antiparallel amide groups can make two pairs of hydrogen bonding with the molecules on the same plane. The odd-spaced bisamide with a parallel directionality of amide groups can form four independent hydrogen bonds with four other bisamide molecules on different planes. The arrangement of bisamide molecules in the crystalline state and the interaction of these molecules depends on the molecular structure, particularly the parity of the spacer length between the amide groups in the bisamide molecule. In this study, the directionality of amide groups has been exploited as a structural characteristic to affect the arrangement of molecules in the crystalline state and produce different binary bisamide gelators with different degrees of crystallinities. Single odd- and even-spaced single bisamides were synthesized and blended to produce binary bisamide organogelators to be characterized in order to understand the effect of the different directionality of amide groups on the molecular interaction in the crystalline state. The pattern of molecular interactions between these blended molecules, mixing or phase separation, has been monitored via differential scanning calorimetry (DSC) and crystallography techniques; X-ray powder diffraction (XRD) and Small-angle X-ray scattering (SAXS). The formation of lamellar structures for odd- and even-spaced bisamide gelators was confirmed by using SAXS and XRD techniques. DSC results have shown that binary bisamide organogelators with different parity of methylene spacers (odd-even binary blends) have a higher tendency for phase separation compared to the binary bisamides with the same parity (odd-odd or even-even binary blends). Phase separation in binary odd-even bisamides was confirmed by the presence of individual (100) reflections of odd and even lamellar structures. The structural characteristic of bisamide organogelators, the parity of spacer length in binary systems, is a promising tool to control the arrangement of molecules and their crystalline structure.

Keywords: binary bisamide organogelators, crystalline structure, phase separation, self-assembly behavior

Procedia PDF Downloads 188
348 Possibilities and Challenges for District Heating

Authors: Louise Ödlund, Danica Djuric Ilic

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From a system perspective, there are several benefits of DH. A possibility to utilize the excess heat from waste incineration and biomass-based combined heat and power (CHP) production (e.g. possibility to utilize the excess heat from electricity production) are two examples. However, in a future sustainable society, the benefits of DH may be less obvious. Due to the climate changes and increased energy efficiency of buildings, the demand for space heating is expected to decrease. Due to the society´s development towards circular economy, a larger amount of the waste will be material recycled, and the possibility for DH production by the energy recovery through waste incineration will be reduced. Furthermore, the benefits of biomass-based CHP production will be less obvious since the marginal electricity production will no longer be linked to high greenhouse gas emissions due to an increased share of renewable electricity capacity in the electricity system. The purpose of the study is (1) to provide an overview of the possible development of other sectors which may influence the DH in the future and (2) to detect new business strategies which would enable for DH to adapt to the future conditions and remain competitive to alternative heat production in the future. A system approach was applied where DH is seen as a part of an integrated system which consists of other sectors as well. The possible future development of other sectors and the possible business strategies for DH producers were searched through a systematic literature review In order to remain competitive to the alternative heat production in the future, DH producers need to develop new business strategies. While the demand for space heating is expected to decrease, the space cooling demand will probably increase due to the climate changes, but also due to the better insulation of buildings in the cases where the home appliances are the heat sources. This opens up a possibility for applying DH-driven absorption cooling, which would increase the annual capacity utilization of the DH plants. The benefits of the DH related to the energy recovery from the waste incineration will exist in the future since there will always be a need to take care of materials and waste that cannot be recycled (e.g. waste containing organic toxins, bacteria, such as diapers and hospital waste). Furthermore, by operating central controlled heat pumps, CHP plants, and heat storage depending on the intermittent electricity production variation, the DH companies may enable an increased share of intermittent electricity production in the national electricity grid. DH producers can also enable development of local biofuel supply chains and reduce biofuel production costs by integrating biofuel and DH production in local DH systems.

Keywords: district heating, sustainable business strategies, sustainable development, system approach

Procedia PDF Downloads 89
347 Li2S Nanoparticles Impact on the First Charge of Li-ion/Sulfur Batteries: An Operando XAS/XES Coupled With XRD Analysis

Authors: Alice Robba, Renaud Bouchet, Celine Barchasz, Jean-Francois Colin, Erik Elkaim, Kristina Kvashnina, Gavin Vaughan, Matjaz Kavcic, Fannie Alloin

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With their high theoretical energy density (~2600 Wh.kg-1), lithium/sulfur (Li/S) batteries are highly promising, but these systems are still poorly understood due to the complex mechanisms/equilibria involved. Replacing S8 by Li2S as the active material allows the use of safer negative electrodes, like silicon, instead of lithium metal. S8 and Li2S have different conductivity and solubility properties, resulting in a profoundly changed activation process during the first cycle. Particularly, during the first charge a high polarization and a lack of reproducibility between tests are observed. Differences observed between raw Li2S material (micron-sized) and that electrochemically produced in a battery (nano-sized) may indicate that the electrochemical process depends on the particle size. Then the major focus of the presented work is to deepen the understanding of the Li2S material charge mechanism, and more precisely to characterize the effect of the initial Li2S particle size both on the mechanism and the electrode preparation process. To do so, Li2S nanoparticles were synthetized according to two ways: a liquid path synthesis and a dissolution in ethanol, allowing Li2S nanoparticles/carbon composites to be made. Preliminary chemical and electrochemical tests show that starting with Li2S nanoparticles could effectively suppress the high initial polarization but also influence the electrode slurry preparation. Indeed, it has been shown that classical formulation process - a slurry composed of Polyvinylidone Fluoride polymer dissolved in N-methyle-2-pyrrolidone - cannot be used with Li2S nanoparticles. This reveals a complete different Li2S material behavior regarding polymers and organic solvents when going at the nanometric scale. Then the coupling between two operando characterizations such as X-Ray Diffraction (XRD) and X-Ray Absorption and Emission Spectroscopy (XAS/XES) have been carried out in order to interpret the poorly understood first charge. This study discloses that initial particle size of the active material has a great impact on the working mechanism and particularly on the different equilibria involved during the first charge of the Li2S based Li-ion batteries. These results explain the electrochemical differences and particularly the polarization differences observed during the first charge between micrometric and nanometric Li2S-based electrodes. Finally, this work could lead to a better active material design and so to more efficient Li2S-based batteries.

Keywords: Li-ion/Sulfur batteries, Li2S nanoparticles effect, Operando characterizations, working mechanism

Procedia PDF Downloads 270
346 Polypeptide Modified Carbon Nanotubes – Mediated GFP Gene Transfection for H1299 Cells and Toxicity Assessment

Authors: Pei-Ying Lo, Jing-Hao Ciou, Kai-Cheng Yang, Jia-Huei Zheng, Shih-Hsiang Huang, Kuen-Chan Lee, Er-Chieh Cho

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As-produced CNTs are insoluble in all organic solvents and aqueous solutions have imposed limitations to the use of CNTs. Therefore, how to debundle carbon nanotubes and to modify them for further uses is an important issue. There are several methods for the dispersion of CNTs in water using covalent attachment of hydrophilic groups to the surface of tubes. These methods, however, alter the electronic structure of the nanotubes by disrupting the network of sp2 hybridized carbons. In order to keep the nanotubes’ intrinsic mechanical and electrical properties intact, non-covalent interactions are increasingly being explored as an alternative route for dispersion. Apart from conventional surfactants such as sodium dodecylsulfate (SDS) or sodium dodecylbenzenesulfonate (SDBS) which are highly effective in dispersing CNTs, biopolymers have received much attention as dispersing agents due to the anticipated biocompatibility of the dispersed CNTs. Also, The pyrenyl group is known to interact strongly with the basal plane of graphene via π-stacking. In this study, a highly re-dispersible biopolymer is reported for the synthesis of pyrene-modified poly-L-lysine (PBPL) and poly(D-Glu, D-Lys) (PGLP). To provide the evidence of the safety of the PBPL/CNT & PGLP/CNT materials we use in this study, H1299 and HCT116 cells were incubated with PBPL/CNT & PGLP/CNT materials for toxicity analysis, MTS assays. The results from MTS assays indicated that no significant cellular toxicity was shown in H1299 and HCT116 cells. Furthermore, the fluorescence marker fluorescein isothiocyanate (FITC) was added to PBPL & PGLP dispersions. From the fluorescent measurements showed that the chemical functionalisation of the PBPL/CNT & PGLP/CNT conjugates with the fluorescence marker were successful. The fluorescent PBPL/CNT & PGLP/CNT conjugates could find application in medical imaging. In the next step, the GFP gene is immobilized onto PBPL/CNT conjugates by introducing electrostatic interaction. GFP-transfected cells that emitted fluorescence were imaged and counted under a fluorescence microscope. Due to the unique biocompatibility of PBPL modified CNTs, the GFP gene could be transported into H1299 cells without using antibodies. The applicability of such soluble and chemically functionalised polypeptide/CNT conjugates in biomedicine is currently investigated. We expect that this polypeptide/CNT system will be a safe and multi-functional nanomedical delivery platform and contribute to future medical therapy.

Keywords: carbon nanotube, nanotoxicology, GFP transfection, polypeptide/CNT hybrids

Procedia PDF Downloads 343
345 Analytical Tools for Multi-Residue Analysis of Some Oxygenated Metabolites of PAHs (Hydroxylated, Quinones) in Sediments

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

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

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

Procedia PDF Downloads 290
344 Use of Geoinformatics and Mathematical Equations to Assess Erosion and Soil Fertility in Cassava Growing Areas in Maha Sarakham Province, Thailand

Authors: Sasirin Srisomkiew, Sireewan Ratsadornasai, Tanomkwan Tipvong, Isariya Meesing

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Cassava is an important food source in the tropics and has recently gained attention as a potential source of biofuel that can replace limited fossil fuel sources. As a result, the demand for cassava production to support industries both within the country and abroad has increased. In Thailand, most farmers prefer to grow cassava in sandy and sandy loam areas where the soil has low natural fertility. Cassava is a tuber plant that has large roots to store food, resulting in the absorption of large amounts of nutrients from the soil, such as nitrogen, phosphorus, and potassium. Therefore, planting cassava in the same area for a long period causes soil erosion and decreases soil fertility. The loss of soil fertility affects the economy, society, and food and energy security of the country. Therefore, it is necessary to know the level of soil fertility and the amount of nutrients in the soil. To address this problem, this study applies geo-informatics technology and mathematical equations to assess erosion and soil fertility and to analyze factors affecting the amount of cassava production in Maha Sarakham Province. The results show that the area for cassava cultivation has increased in every district of Maha Sarakham Province between 2015-2022, with the total area increasing to 180,922 rai or 5.47% of the province’s total area during this period. Furthermore, it was found that it is possible to assess areas with soil erosion problems that had a moderate level of erosion in areas with high erosion rates ranging from 5-15 T/rai/year. Soil fertility assessment and information obtained from the soil nutrient map for 2015–2023 reveal that farmers in the area have improved the soil by adding chemical fertilizers along with organic fertilizers, such as manure and green manure, to increase the amount of nutrients in the soil. This is because the soil resources of Maha Sarakham Province mostly have relatively low agricultural potential due to the soil texture being sand and sandy loam. In this scenario, the ability to absorb nutrients is low, and the soil holds little water, so it is naturally low in fertility. Moreover, agricultural soil problems were found, including the presence of saline soil, sandy soil, and acidic soil, which is a serious restriction on land use because it affects the release of nutrients into the soil. The results of this study may be used as a guideline for managing soil resources and improving soil quality to prevent soil degradation problems that may occur in the future.

Keywords: Cassava, geoinformatics, soil erosion, soil fertility, land use change

Procedia PDF Downloads 57
343 Genetic Analysis of CYP11A1 Gene with Polycystic Ovary Syndrome from North India

Authors: Ratneev Kaur, Tajinder Kaur, Anupam Kaur

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Introduction: Polycystic Ovary Syndrome (PCOS) is a heterogenous disorder of endocrine system among women of reproductive age. PCOS is characterized by hyperandrogenism, anovulation, polycystic ovaries, hirsutism, obesity, and hyperinsulinemia. Several pathways are implicated in its etiology including the metabolic pathway of steroid hormone synthesis regulatory pathways. PCOS is an androgen excess disorder, genes operating in steroidogenesis may alter pathogenesis of PCOS. The cytochrome P450scc is a cholesterol side chain cleavage enzyme coded by CYP11A1 gene and catalyzes conversion of cholesterol to pregnenolone, the initial and rate-limiting step in steroid hormone synthesis. It is postulated that polymorphisms in this gene may play an important role in the regulation of CYP11A1 expression and leading to increased or decreased androgen production. The present study will be the first study from north India to best of our knowledge, to analyse the association of CYP11A1 (rs11632698) polymorphism in women suffering from PCOS. Methodology: The present study was approved by ethical committee of Guru Nanak Dev University in consistent with declaration of Helsinki. A total of 300 samples (150 PCOS cases and 150 controls) were recruited from Hartej hospital, for the present study. Venous blood sample (3ml) was withdrawn from women diagnosed with PCOS by doctor, according to Rotterdam 2003 criteria and from healthy age matched controls only after informed consent and detailed filled proforma. For molecular genetics analysis, blood was stored in EDTA vials. After DNA isolation by organic method, PCR-RFLP approach was used for genotyping and association analysis of rs11632698 polymorphism. Statistical analysis was done to check for significance of selected polymorphism with PCOS. Results: In 150 PCOS cases, the frequency of AA, AG and GG genotype was found to be 48%, 35%, and 13% compared to 62%, 27% and 8% in 150 controls. The major allele (A) and minor allele (G) frequency was 68% and 32% in cases and 78% and 22% in controls. Minor allele frequency was higher in cases as compared to controls, as well as the distribution of genotype was observed to be statistically significant (ᵡ²=6.525, p=0.038). Odds ratio in dominant, co-dominant and recessive models observed was 1.81 (p=0.013), 1.54 (p=0.012) and 1.77 (p=0.132) respectively. Conclusion: The present study showed statistically significant association of rs11632698 with PCOS (p=0.038) in North Indian women.

Keywords: polycystic ovary syndrome, CYP11A1, rs11632698, hyperandrogenism

Procedia PDF Downloads 146
342 The Composition of Biooil during Biomass Pyrolysis at Various Temperatures

Authors: Zoltan Sebestyen, Eszter Barta-Rajnai, Emma Jakab, Zsuzsanna Czegeny

Abstract:

Extraction of the energy content of lignocellulosic biomass is one of the possible pathways to reduce the greenhouse gas emission derived from the burning of the fossil fuels. The application of the bioenergy can mitigate the energy dependency of a country from the foreign natural gas and the petroleum. The diversity of the plant materials makes difficult the utilization of the raw biomass in power plants. This problem can be overcome by the application of thermochemical techniques. Pyrolysis is the thermal decomposition of the raw materials under inert atmosphere at high temperatures, which produces pyrolysis gas, biooil and charcoal. The energy content of these products can be exploited by further utilization. The differences in the chemical and physical properties of the raw biomass materials can be reduced by the use of torrefaction. Torrefaction is a promising mild thermal pretreatment method performed at temperatures between 200 and 300 °C in an inert atmosphere. The goal of the pretreatment from a chemical point of view is the removal of water and the acidic groups of hemicelluloses or the whole hemicellulose fraction with minor degradation of cellulose and lignin in the biomass. Thus, the stability of biomass against biodegradation increases, while its energy density increases. The volume of the raw materials decreases so the expenses of the transportation and the storage are reduced as well. Biooil is the major product during pyrolysis and an important by-product during torrefaction of biomass. The composition of biooil mostly depends on the quality of the raw materials and the applied temperature. In this work, thermoanalytical techniques have been used to study the qualitative and quantitative composition of the pyrolysis and torrefaction oils of a woody (black locust) and two herbaceous samples (rape straw and wheat straw). The biooil contains C5 and C6 anhydrosugar molecules, as well as aromatic compounds originating from hemicellulose, cellulose, and lignin, respectively. In this study, special emphasis was placed on the formation of the lignin monomeric products. The structure of the lignin fraction is different in the wood and in the herbaceous plants. According to the thermoanalytical studies the decomposition of lignin starts above 200 °C and ends at about 500 °C. The lignin monomers are present among the components of the torrefaction oil even at relatively low temperatures. We established that the concentration and the composition of the lignin products vary significantly with the applied temperature indicating that different decomposition mechanisms dominate at low and high temperatures. The evolutions of decomposition products as well as the thermal stability of the samples were measured by thermogravimetry/mass spectrometry (TG/MS). The differences in the structure of the lignin products of woody and herbaceous samples were characterized by the method of pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS). As a statistical method, principal component analysis (PCA) has been used to find correlation between the composition of lignin products of the biooil and the applied temperatures.

Keywords: pyrolysis, torrefaction, biooil, lignin

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341 Evaluation of Antimicrobial and Anti-Inflammatory Activity of Doani Sidr Honey and Madecassoside against Propionibacterium Acnes

Authors: Hana Al-Baghaoi, Kumar Shiva Gubbiyappa, Mayuren Candasamy, Kiruthiga Perumal Vijayaraman

Abstract:

Acne is a chronic inflammatory disease of the sebaceous glands characterized by areas of skin with seborrhea, comedones, papules, pustules, nodules, and possibly scarring. Propionibacterium acnes (P. acnes), plays a key role in the pathogenesis of acne. Their colonization and proliferation trigger the host’s inflammatory response leading to the production of pro-inflammatory cytokines such as interleukin-8 (IL-8) and tumour necrosis factor-α (TNF-α). The usage of honey and natural compounds to treat skin ailments has strong support in the current trend of drug discovery. The present study was carried out evaluate antimicrobial and anti-inflammatory potential of Doani Sidr honey and its fractions against P. acnes and to screen madecassoside alone and in combination with fractions of honey. The broth dilution method was used to assess the antibacterial activity. Also, ultra structural changes in cell morphology were studied before and after exposure to Sidr honey using transmission electron microscopy (TEM). The three non-toxic concentrations of the samples were investigated for suppression of cytokines IL 8 and TNF α by testing the cell supernatants in the co-culture of the human peripheral blood mononuclear cells (hPBMCs) heat killed P. acnes using enzyme immunoassay kits (ELISA). Results obtained was evaluated by statistical analysis using Graph Pad Prism 5 software. The Doani Sidr honey and polysaccharide fractions were able to inhibit the growth of P. acnes with a noteworthy minimum inhibitory concentration (MIC) value of 18% (w/v) and 29% (w/v), respectively. The proximity of MIC and MBC values indicates that Doani Sidr honey had bactericidal effect against P. acnes which is confirmed by TEM analysis. TEM images of P. acnes after treatment with Doani Sidr honey showed completely physical membrane damage and lysis of cells; whereas non honey treated cells (control) did not show any damage. In addition, Doani Sidr honey and its fractions significantly inhibited (> 90%) of secretion of pro-inflammatory cytokines like TNF α and IL 8 by hPBMCs pretreated with heat-killed P. acnes. However, no significant inhibition was detected for madecassoside at its highest concentration tested. Our results suggested that Doani Sidr honey possesses both antimicrobial and anti-inflammatory effects against P. acnes and can possibly be used as therapeutic agents for acne. Furthermore, polysaccharide fraction derived from Doani Sidr honey showed potent inhibitory effect toward P. acnes. Hence, we hypothesize that fraction prepared from Sidr honey might be contributing to the antimicrobial and anti-inflammatory activity. Therefore, this polysaccharide fraction of Doani Sidr honey needs to be further explored and characterized for various phytochemicals which are contributing to antimicrobial and anti-inflammatory properties.

Keywords: Doani sidr honey, Propionibacterium acnes, IL-8, TNF alpha

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340 Sorghum Polyphenols Encapsulated by Spray Drying, Using Modified Starches as Wall Materials

Authors: Adriana Garcia G., Alberto A. Escobar P., Amira D. Calvo L., Gabriel Lizama U., Alejandro Zepeda P., Fernando Martínez B., Susana Rincón A.

Abstract:

Different studies have recently been focused on the use of antioxidants such as polyphenols because of to its anticarcinogenic capacity. However, these compounds are highly sensible to environmental factors such as light and heat, so lose its long-term stability, besides possess an astringent and bitter taste. Nevertheless, the polyphenols can be protected by microcapsule formulation. In this sense, a rich source of polyphenols is sorghum, besides presenting a high starch content. Due to the above, the aim of this work was to obtain modified starches from sorghum by extrusion to encapsulate polyphenols the sorghum by spray drying. Polyphenols were extracted by ethanol solution from sorghum (Pajarero/red) and determined by the method of Folin-Ciocalteu, obtaining GAE at 30 mg/g. Moreover, was extracted starch of sorghum (Sinaloense/white) through wet milling (yield 32 %). The hydrolyzed starch was modified with three treatments: acetic anhydride (2.5g/100g), sodium tripolyphosphate (4g/100g), and sodium tripolyphosphate/ acetic anhydride (2g/1.25g by each 100 g) by extrusion. Processing conditions of extrusion were as follows: barrel temperatures were of 60, 130 and 170 °C at the feeding, transition, and high-pressure extrusion zones, respectively. Analysis of Fourier Transform Infrared spectroscopy (FTIR), showed bands exhibited of acetyl groups (1735 cm-1) and phosphates (1170 cm-1, 910 cm-1 and 525 cm-1), indicating the respective modification of starch. Besides, all modified starches not developed viscosity, which is a characteristic required for use in the encapsulation of polyphenols using the spray drying technique. As result of the modification starch, was obtained a water solubility index (WSI) from 33.8 to 44.8 %, and crystallinity from 8 to 11 %, indicating the destruction of the starch granule. Afterwards, microencapsulation of polyphenols was developed by spray drying, with a blend of 10 g of modified starch, 60 ml polyphenol extract and 30 ml of distilled water. Drying conditions were as follows: inlet air temperature 150 °C ± 1, outlet air temperature 80°C ± 5. As result of the microencapsulation: were obtained yields of 56.8 to 77.4 % and an efficiency of encapsulation from 84.6 to 91.4 %. The FTIR analysis showed evidence of microcapsules loaded with polyphenols in bands 1042 cm-1, 1038 cm-1 and 1148 cm-1. Analysis Differential scanning calorimetry (DSC) showed transition temperatures from 144.1 to 173.9 °C. For the order hand, analysis of Scanning Electron Microscopy (SEM), were observed rounded surfaces with concavities, typical feature of microcapsules produced by spray drying, how result of rapid evaporation of water. Finally, the modified starches were obtained by extrusion with good characteristics for use as cover materials by spray drying, where the phosphorylated starch was the best treatment in this work, according to the encapsulation yield, efficiency, and transition temperature.

Keywords: encapsulation, extrusion, modified starch, polyphenols, spray drying

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339 Cytotoxic Effect of Biologically Transformed Propolis on HCT-116 Human Colon Cancer Cells

Authors: N. Selvi Gunel, L. M. Oktay, H. Memmedov, B. Durmaz, H. Kalkan Yildirim, E. Yildirim Sozmen

Abstract:

Object: Propolis which consists of compounds that are accepted as antioxidant, antimicrobial, antiseptic, antibacterial, anti-inflammatory, anti-mutagenic, immune-modulator and cytotoxic, is frequently used in current therapeutic applications. However, some of them result in allergic side effects, causing consumption to be restricted. Previously our group has succeeded in producing a new biotechnological product which was less allergenic. In this study, we purpose to optimize production conditions of this biologically-transformed propolis and determine the cytotoxic effects of obtained new products on colon cancer cell line (HCT-116). Method: Firstly, solid propolis samples were dissolved in water after weighing, grinding and sizing (sieve-35mesh) and applied 40 kHz/10 min ultrasonication. Samples were prepared according to inoculation with Lactobacillus plantarum in two different proportions (2.5% and 3.5%). Chromatographic analyzes of propolis were performed by UPLC-MS/MS (Waters, Milford, MA) system. Results were analysed by UPLC-MS/MS system MassLynx™ 4.1 software. HCT-116 cells were treated with propolis examples at 25-1000 µg/ml concentrations and cytotoxicity were measured by using WST-8 assay at 24, 48, and 72 hours. Samples with biological transformation were compared with the non-transformed control group samples. Our experiment groups were formed as follows: untreated (group 1), propolis dissolved in water ultrasonicated at 40 kHz/10 min (group 2), propolis dissolved in water ultrasonicated at 40 kHz/10 min and inoculated 2.5% L. plantarum L1 strain (group 3), propolis dissolved in water ultrasonicated at 40 kHz/10 min and inoculated 3.5% L. plantarum L3 strain (group 4). Obtained data were calculated with Graphpad Software V5 and analyzed by two-way ANOVA test followed by Bonferroni test. Result: As a result of our study, the cytotoxic effect of propolis samples on HCT-116 cells was evaluated. There was a 7.21 fold increase in group 3 compared to group 2 in the concentration of 1000 µg/ml, and it was a 6.66 fold increase in group 3 compared to group 1 at the end of 24 hours. At the end of 48 hours, in the concentration of 500 µg/ml, it was determined 4.7 fold increase in group 4 compared to group 3. At the same time, in the concentration of 750 µg/ml it was determined 2.01 fold increase in group 4 compared to group 3 and in the same concentration, it was determined 3.1 fold increase in group 4 compared to group 2. Also, at the 72 hours, in the concentration of 750 µg/ml, it was determined 2.42 fold increase in group 3 according to group 2 and in the same time, in the concentration of 1000 µg/ml, it was determined 2.13 fold increase in group 4 according to group 2. According to cytotoxicity results, the group which were ultrasonicated at 40 kHz/10min and inoculated 3.5% L. plantarum L3-strain had a higher cytotoxic effect. Conclusion: It is known that bioavailability of propolis is halved in six months. The data obtained from our results indicated that biologically-transformed propolis had more cytotoxic effect than non-transformed group on colon cancer cells. Consequently, we suggested that L. plantarum-transformation provides both reduction of allergenicity and extension of bioavailability period by enhancing healthful polyphenols.

Keywords: bio-transformation, propolis, colon cancer, cytotoxicity

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338 High Throughput Virtual Screening against ns3 Helicase of Japanese Encephalitis Virus (JEV)

Authors: Soma Banerjee, Aamen Talukdar, Argha Mandal, Dipankar Chaudhuri

Abstract:

Japanese Encephalitis is a major infectious disease with nearly half the world’s population living in areas where it is prevalent. Currently, treatment for it involves only supportive care and symptom management through vaccination. Due to the lack of antiviral drugs against Japanese Encephalitis Virus (JEV), the quest for such agents remains a priority. For these reasons, simulation studies of drug targets against JEV are important. Towards this purpose, docking experiments of the kinase inhibitors were done against the chosen target NS3 helicase as it is a nucleoside binding protein. Previous efforts regarding computational drug design against JEV revealed some lead molecules by virtual screening using public domain software. To be more specific and accurate regarding finding leads, in this study a proprietary software Schrödinger-GLIDE has been used. Druggability of the pockets in the NS3 helicase crystal structure was first calculated by SITEMAP. Then the sites were screened according to compatibility with ATP. The site which is most compatible with ATP was selected as target. Virtual screening was performed by acquiring ligands from databases: KinaseSARfari, KinaseKnowledgebase and Published inhibitor Set using GLIDE. The 25 ligands with best docking scores from each database were re-docked in XP mode. Protein structure alignment of NS3 was performed using VAST against MMDB, and similar human proteins were docked to all the best scoring ligands. The low scoring ligands were chosen for further studies and the high scoring ligands were screened. Seventy-three ligands were listed as the best scoring ones after performing HTVS. Protein structure alignment of NS3 revealed 3 human proteins with RMSD values lesser than 2Å. Docking results with these three proteins revealed the inhibitors that can interfere and inhibit human proteins. Those inhibitors were screened. Among the ones left, those with docking scores worse than a threshold value were also removed to get the final hits. Analysis of the docked complexes through 2D interaction diagrams revealed the amino acid residues that are essential for ligand binding within the active site. Interaction analysis will help to find a strongly interacting scaffold among the hits. This experiment yielded 21 hits with the best docking scores which could be investigated further for their drug like properties. Aside from getting suitable leads, specific NS3 helicase-inhibitor interactions were identified. Selection of Target modification strategies complementing docking methodologies which can result in choosing better lead compounds are in progress. Those enhanced leads can lead to better in vitro testing.

Keywords: antivirals, docking, glide, high-throughput virtual screening, Japanese encephalitis, ns3 helicase

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