Search results for: adipocyte-fatty acid binding protein

Authors: Iryna Atamaniuk, Hannah Boysen, Nils Wieczorek, Natalia Politaeva, Iuliia Bazarnova, Kerstin Kuchta

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Over the last decade due to climate change and a strategy of natural resources preservation, the interest for the aquatic biomass has dramatically increased. Along with mitigation of the environmental pressure and connection of waste streams (including CO₂ and heat emissions), microalgae bioeconomy can supply food, feed, as well as the pharmaceutical and power industry with number of value-added products. Furthermore, in comparison to conventional biomass, microalgae can be cultivated in wide range of conditions without compromising food and feed production, thus addressing issues associated with negative social and the environmental impacts. This paper presents the state-of-the art technology for microalgae bioeconomy from cultivation process to production of valuable components and by-streams. Microalgae Chlorella sorokiniana were cultivated in the pilot-scale innovation concept in Hamburg (Germany) using different systems such as race way pond (5000 L) and flat panel reactors (8 x 180 L). In order to achieve the optimum growth conditions along with suitable cellular composition for the further extraction of the value-added components, process parameters such as light intensity, temperature and pH are continuously being monitored. On the other hand, metabolic needs in nutrients were provided by addition of micro- and macro-nutrients into a medium to ensure autotrophic growth conditions of microalgae. The cultivation was further followed by downstream process and extraction of lipids, proteins and saccharides. Lipids extraction is conducted in repeated-batch semi-automatic mode using hot extraction method according to Randall. As solvents hexane and ethanol are used at different ratio of 9:1 and 1:9, respectively. Depending on cell disruption method along with solvents ratio, the total lipids content showed significant variations between 8.1% and 13.9 %. The highest percentage of extracted biomass was reached with a sample pretreated with microwave digestion using 90% of hexane and 10% of ethanol as solvents. Proteins content in microalgae was determined by two different methods, namely: Total Kejadahl Nitrogen (TKN), which further was converted to protein content, as well as Bradford method using Brilliant Blue G-250 dye. Obtained results, showed a good correlation between both methods with protein content being in the range of 39.8–47.1%. Characterization of neutral and acid saccharides from microalgae was conducted by phenol-sulfuric acid method at two wavelengths of 480 nm and 490 nm. The average concentration of neutral and acid saccharides under the optimal cultivation conditions was 19.5% and 26.1%, respectively. Subsequently, biomass residues are used as substrate for anaerobic digestion on the laboratory-scale. The methane concentration, which was measured on the daily bases, showed some variations for different samples after extraction steps but was in the range between 48% and 55%. CO₂ which is formed during the fermentation process and after the combustion in the Combined Heat and Power unit can potentially be used within the cultivation process as a carbon source for the photoautotrophic synthesis of biomass.

Keywords: bioeconomy, lipids, microalgae, proteins, saccharides

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Authors: Dalia. G. Aseel, E. E. Hafez, S. M. Hammad

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Viral RNAs of both potato leaf roll virus (PLRV) and potato virus Y (PVY) were extracted from infected potato leaves collected from different Egyptian regions. Differential Display Polymerase Chain Reaction (DD-PCR) using (Endogluconase, β-1,3-glucanases, Chitinase, Peroxidase and Polyphenol oxidase) primers (forward strand) for was performed. The obtained data revealed different banding patterns depending on the viral type and the region of infection. Regarding PLRV, a 58 up regulated and 19 down regulated genes were detected, while, 31 up regulated and 14 down regulated genes were observed in case of PVY. Based on the nucleotide sequencing, variable phylogenetic relationships were reported for the three sequenced genes coding for: Induced stolen tip protein, Disease resistance RPP-like protein and non-specific lipid-transfer protein. In a complementary approach, using the quantitative Real-time PCR, the expressions of PRs genes understudy were estimated in the infected leaves by PLRV and PVY of three potato cultivars (Spunta, Diamont and Cara). The infection with both viruses inhibited the expressions of the five PRs genes. On the contrary, infected leaves by PLRV or PVY elevated the expression of some defense genes. This interaction also may be enhanced and/or inhibited the expression of some genes responsible for the plant defense mechanisms.

Keywords: PLRV, PVY, PR genes, DD-PCR, qRT-PCR, sequencing

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Authors: Mourad Makhlouf, Omar Bouchher, Messabih Sidi Mohamed, Benrachedi Khaled

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The objective of this study was to investigate the removal of two organic pollutants; 4-hydroxybenzoic acid (p-hydroxybenzoic acid) and phenol from synthetic wastewater by the adsorption on mesoporous materials. In this context, the aim of this work is to study the adsorption of organic compounds phenol and 4AHB on MCM-41 and FSM-16 non-grafted (NG) and other grafted (G) by trimethylchlorosilane (TMCS). The results of phenol and 4AHB adsorption in aqueous solution show that the adsorption capacity tends to increase after grafting in relation to the increase in hydrophobicity. The materials are distinguished by a higher adsorption capacity to the other NG materials. The difference in the phenol is 14.43% (MCM-41), 14.55% (FSM-16), and 16.72% (MCM-41), 13.57% (FSM-16) in the 4AHB. Our adsorption results show that the grafted materials by TMCS are good adsorbent at 25 °C.

Keywords: MCM-41, FSM-16, TMCS, phenol, 4AHB

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Authors: Ekta, G. K. Darbha

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Nanotechnology offers the potential of simultaneously increasing efficiency as compared to their bulk material as well as reducing harmful environmental impacts of pesticides in field of agriculture. The term nanopesticide covers different pesticides that are cumulative of several surfactants, polymers, metal ions, etc. of nanometer size ranges from 1-1000 nm and exhibit abnormal behavior (high efficacy and high specific surface area) of nanomaterials. Commercial formulations of pesticides used by farmers nowadays cannot be used effectively due to a number of problems associated with them. For example, more than 90% of applied formulations are either lost in the environment or unable to reach the target area required for effective pest control. Around 20−30% of pesticides are lost through emissions. A number of factors (application methods, physicochemical properties of the formulations, and environmental conditions) can influence the extent of loss during application. It is known that among various formulations, polymer-based formulations show the greatest potential due to their greater efficacy, slow release and protection against premature degradation of active ingredient as compared to other commercial formulations. However, the nanoformulations can have a significant effect on the fate of active ingredient as well as may release some new ingredients by reacting with existing soil contaminants. Environmental fate of these newly generated species is still not explored very well which is essential to field scale experiments and hence a lot to be explored in the field of environmental fate, nanotoxicology, transport properties and stability of such formulations. In our preliminary work, we have synthesized polymer based nanoformulation of commercially used weedicide atrazine. Atrazine belongs to triazine class of herbicide, which is used in the effective control of seed germinated dicot weeds and grasses. It functions by binding to the plastoquinone-binding protein in PS-II. Plant death results from starvation and oxidative damage caused by breakdown in electron transport system. The stability of the suspension of nanoformulation containing herbicide has been evaluated by considering different parameters like polydispersity index, particle diameter, zeta-potential under different environmental relevance condition such as pH range 4-10, temperature range from 25°C to 65°C and stability of encapsulation also have been studied for different amount of added polymer. Morphological characterization has been done by using SEM.

Keywords: atrazine, nanoformulation, nanopesticide, nanotoxicology

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Authors: Cindy Woods

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After years of implacable neoliberal globalization, multinational corporations have moved from the periphery to the center of the international legal agenda. Human rights advocates have long called for greater corporate accountability in the international arena. The creation of the Global Compact in 2000, while aimed at fostering greater corporate respect for human rights, did not silence these calls. After multiple unsuccessful attempts to adopt a set of norms relating to the human rights responsibilities of transnational corporations, the United Nations succeeded in 2008 with the Guiding Principles on Business and Human Rights (Guiding Principles). The Guiding Principles, praised by some within the international human rights community for their recognition of an individual corporate responsibility to respect human rights, have not escaped their share of criticism. Many view the Guiding Principles to be toothless, failing to directly impose obligations upon corporations, and call for binding international obligations on corporate entities. After decades of attempting to promulgate human rights obligations for multinational corporations, the existing legal frameworks in place fall short of protecting individuals from the human rights abuses of multinational corporations. The Global Compact and Guiding Principles are proof of the United Nations’ unwillingness to impose international legal obligations on corporate actors. In June 2014, the Human Rights Council adopted a resolution to draft international legally binding human rights norms for business entities; however, key players in the international arena have already announced they will not cooperate with such efforts. This Note, through an overview of the existing corporate accountability frameworks and a study of Newmont Mining’s Minas Conga project in Peru, argues that binding international human rights obligations on corporations are necessary to fully protect human rights. Where states refuse to or simply cannot uphold their duty to protect individuals from transnational businesses’ human rights transgressions, there must exist mechanisms to pursue justice directly against the multinational corporation.

Keywords: business and human rights, Latin America, international treaty on business and human rights, mining, human rights

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Authors: Debarun Dhar Purkayastha, Talinungsang

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SnO₂-TiO₂ nanocomposite thin films were prepared by the sol-gel method on borosilicate glass substrate. The films were annealed at a temperature of 300ᵒC, 400ᵒC, and 500ᵒC respectively for 2h in the air. The films obtained were further modified with stearic acid in order to decrease the surface energy. The X-ray diffraction patterns for the SnO₂-TiO₂ thin films after annealing at different temperatures can be indexed to the mixture of TiO₂ (rutile and anatase) and SnO₂ (tetragonal) phases. The average crystallite size calculated from Scherrer’s formula is found to be 6 nm. The SnO₂-TiO₂ thin films were hydrophilic which on modification with stearic acid exhibit superhydrophobic behavior. The increase in hydrophobicity of SnO₂ film with stearic acid modification is attributed to the change in surface energy of the film. The films exhibit superhydrophilic behavior under UV irradiation for 1h. Thus, it is observed that stearic acid modified surfaces are superhydrophobic but convert into superhydrophilic on being subjected to UV irradiation. SnO₂-TiO₂ thin films have potential for self-cleaning applications because of photoinduced hydrophilicity under UV irradiation.

Keywords: nanocomposite, self-cleaning, superhydrophobic, surface energy

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Authors: Bingjie Wang, Su-Yeon Kwon, Ik-Joong Kang

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The goal of this experiment is to develop a sensor that can quickly check the concentration by using the nanoparticles made by chitosan and gold. Using chitosan nanoparticles crosslinking with sodium tripolyphosphate(TPP) is the first step to form the chitosan nanoparticles, which would be covered with the gold sequentially. The size of the fabricated product was around 100nm. Based on the method that the sulfur end of the MPA linked to gold can form the very strong S–Au bond, and the carboxyl group, the other end of the MPA, can easily absorb the GABA. As for the GABA, what is the primary inhibitory neurotransmitter in the mammalian central nervous system in the human body. It plays such significant role in reducing neuronal excitability pass through the nervous system. A Surface-enhanced Raman Scattering (SERS) as the principle for enhancing Raman scattering by molecules adsorbed on rough metal surfaces or by nanostructures is used to detect the concentration change of γ-Aminobutyric Acid (GABA). When the system is formed, it generated SERS, which made a clear difference in the intensity of Raman scattering within the range of GABA concentration. So it is obtained from the experiment that the calibration curve according to the GABA concentration relevant with the SERS scattering. In this study, DLS, SEM, FT-IR, UV, SERS were used to analyze the products to obtain the conclusion.

Keywords: mercaptopropionic acid, chitosan-gold nanoshell, γ-aminobutyric acid, surface-enhanced raman scattering

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Authors: C. Gallottini, W. Di Mari, C. De Carolis, A. Dolci, G. Dolci, L. Gallottini, G. Barraco, S. Eramo

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The aim of this paper is to summarize the literature on micromorphology and composition of the enamel of the cat and present an original experiment by SEM on how it responds to the etching with ortophosphoric acid for the time recommended in the veterinary literature (30", 45", 60"), derived from research and experience on human enamel; 21 teeth of cat were randomly divided into three groups of 7 (A, B, C): Group A was subjected to etching for 30 seconds by means of orthophosphoric acid to 40% on a circular area with diameter of about 2mm of the enamel coronal; the Groups B and C had the same treatment but, respectively, for 45 and 60 seconds. The samples obtained were observed by SEM to constant magnification of 1000x framing, in particular, the border area between enamel exposed and not exposed to etching to highlight differences. The images were subjected to the analysis of three blinded experienced operators in electron microscopy. In the enamel of the cat the etching for the times considered is not optimally effective for the purpose adhesives and the presence of a thick prismless layer could explain this situation. To improve this condition may clinically in the likeness of what is proposed for the enamel of human deciduous teeth: a bevel or a chamfer of 1 mm on the contour of the cavity to discover the prismatic enamel and increase the bonding surface.

Keywords: cat enamel, SEM, veterinary dentistry, acid etching

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Authors: G. Hosseini, A. Sadighzadeh, M. Rahimnejad, N. Hosseini, Z. Jamalzadeh

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Metal pollutants, especially heavy metals from anthropogenic sources such as metallurgical industries’ waste including mining, smelting, casting or production of nuclear fuel, including mining, concentrate production and uranium processing ends in the environment contamination (water and soil) and risk to human health around the facilities of this type of industrial activity. There are different methods that can be used to remove these contaminants from water and soil. These are very expensive and time-consuming. In this case, the people have been forced to leave the area and the decontamination is not done. For example, in the case of Chernobyl accident, an area of 30 km around the plant was emptied of human life. A very efficient and cost-effective method for decontamination of the soil and the water is phytoremediation. In this method, the plants preferentially native plants which are more adaptive to the regional climate are well used. In this study, three types of plants including Alfalfa, Sunflower and wheat were used to Barium decontamination. Alfalfa and Sunflower were not grown good enough in Saghand mine’s soil sample. This can be due to non-native origin of these plants. But, Wheat rise in Saghand Uranium Mine soil sample was satisfactory. In this study, we have investigated the effect of 4 types of acids inclusive nitric acid, oxalic acid, acetic acid and citric acid on the removal efficiency of Barium by Wheat. Our results indicate the increase of Barium absorption in the presence of citric acid in the soil. In this paper, we will present our research and laboratory results.

Keywords: phytoremediation, heavy metal, wheat, soil

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Authors: Joon-Suk Kim, Hae-Woo Lee

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This study discusses the effect of welding heat input on intergranular corrosion of the weld metal of Inconel 625 alloy. A specimen of Inconel 625 with a weld metal that controlled welding heat input was manufactured, and aging heat treatment was conducted to investigate sensitization by chromium carbides. The electrochemical SL and DL EPR experiments, together with the chemical ferric sulfate-sulfuric acid and nitric acid tests, were conducted to determine intergranular corrosion susceptibility between the specimens. In the SL and DL EPR experiments, specimens were stabilized in the weld metal, and therefore intergranular corrosion susceptibility could not be determined. However, in the ferric sulfate-sulfuric acid and nitric acid tests, the corrosion speed increased as heat input increased. This was because the amount of diluted Fe increased as the welding heat input increased, leading to microsegregation between the dendrites, which had a negative effect on the corrosion resistance.

Keywords: Inconel 625, weling, overlay, heat input, intergranular corrosion

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Authors: Olubukayo-Opeyemi Oyetayo, Oscar Mendez-Lucio, Andreas Bender, Hans Kiefer

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The thermal melting curve of a protein provides information on its conformational stability and could provide cues on its aggregation behavior. Naturally-occurring osmolytes have been shown to improve the thermal stability of most proteins in a concentration-dependent manner. They are therefore commonly employed as additives in therapeutic protein purification and formulation. A number of intertwined and seemingly conflicting mechanisms have been put forward to explain the observed stabilizing effects, the most prominent being the preferential exclusion mechanism. We attempted to probe and summarize molecular mechanisms for thermal stabilization of a monoclonal antibody (mAb) by developing quantitative structure-activity relationships using a rationally-selected library of 120 osmolyte-like compounds in the polyhydric alcohols, amino acids and methylamines classes. Thermal stabilization potencies were experimentally determined by thermal shift assays based on differential scanning fluorimetry. The cross-validated QSAR model was developed by partial least squares regression using descriptors generated from Molecular Operating Environment software. Careful evaluation of the results with the use of variable importance in projection parameter (VIP) and regression coefficients guided the selection of the most relevant descriptors influencing mAb thermal stability. For the mAb studied and at pH 7, the thermal stabilization effects of tested compounds correlated positively with their fractional polar surface area and inversely with their fractional hydrophobic surface area. We cannot claim that the observed trends are universal for osmolyte-protein interactions because of protein-specific effects, however this approach should guide the quick selection of (de)stabilizing compounds for a protein from a chemical library. Further work with a large variety of proteins and at different pH values would help the derivation of a solid explanation as to the nature of favorable osmolyte-protein interactions for improved thermal stability. This approach may be beneficial in the design of novel protein stabilizers with optimal property values, especially when the influence of solution conditions like the pH and buffer species and the protein properties are factored in.

Keywords: thermal stability, monoclonal antibodies, quantitative structure-activity relationships, osmolytes

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Authors: Narin Salehiyan, Ramin Ghasemi Shayan

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In order to investigate coronavirus RNA replication, transcription, recombination, protein processing and transport, virion assembly, the identification of coronavirus-specific cell receptors, and polymerase processing, the manipulation of coronavirus clones and complementary DNAs (cDNAs) of defective-interfering (DI) RNAs is the subject of this chapter. The idea of the Covid genome is nonsegmented, single-abandoned, and positive-sense RNA. When compared to other RNA viruses, its size is significantly greater, ranging from 27 to 32 kb. The quality encoding the enormous surface glycoprotein depends on 4.4 kb, encoding a forcing trimeric, profoundly glycosylated protein. This takes off exactly 20 nm over the virion envelope, giving the infection the appearance-with a little creative mind of a crown or coronet. Covid research has added to the comprehension of numerous parts of atomic science as a general rule, like the component of RNA union, translational control, and protein transport and handling. It stays a fortune equipped for creating startling experiences.

Keywords: covid-19, corona, virus, genome, genetic

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Authors: Awf A. Al-Khan, Michael J. Day, Judith Nimmo, Mourad Tayebi, Stewart D. Ryan, Samantha J. Richardson, Janine A. Danks

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Osteosarcoma (OS) is the most common type of malignant primary bone tumour in dogs. In addition to their critical roles in bone formation and remodeling, parathyroid hormone-related protein (PTHrP) and its receptor (PTHR1) are involved in progression and metastasis of many types of tumours in humans. The aims of this study were to determine the localisation and expression levels of PTHrP and PTHR1 in canine OS tissues using immunohistochemistry and to investigate if this expression is correlated with survival time. Formalin-fixed, paraffin-embedded tissue samples from 44 dogs with known survival time that had been diagnosed with primary osteosarcoma were analysed for localisation of PTHrP and PTHR1. Findings showed that both PTHrP and PTHR1 were present in all OS samples. The dogs with high level of PTHR1 protein (16%) had decreased survival time (P<0.05) compared to dogs with less PTHR1 protein. PTHrP levels did not correlate with survival time (P>0.05). The results of this study indicate that the PTHR1 is expressed differently in canine OS tissues and this may be correlated with poor prognosis. This may mean that PTHR1 may be useful as a prognostic indicator in canine OS and could represent a good therapeutic target in OS.

Keywords: dog, expression, osteosarcoma, parathyroid hormone receptor 1 (PTHR1), parathyroid hormone-related protein (PTHrP), survival

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Authors: Ibrahim Iddrisu, Joseph Ayariga, Junhuan Xu, Ayomide Adebanjo, Boakai K. Robertson, Michelle Samuel-Foo, Olufemi Ajayi

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The rise of antimicrobial resistance is a global public health crisis that threatens the effective control and prevention of infections. Due to the emergence of pan drug-resistant bacteria, most antibiotics have lost their efficacy. Bacteriophages or their components are known to target bacterial cell walls, cell membranes, and lipopolysaccharides (LPS) and hydrolyze them. Bacteriophages, being the natural predators of pathogenic bacteria, are inevitably categorized as ‘human friends’, thus fulfilling the adage that ‘the enemy of my enemy is my friend’. Leveraging on their lethal capabilities against pathogenic bacteria, researchers are searching for more ways to overcome the current antibiotic resistance challenge. In this study, we expressed and purified epsilon 34 phage tail spike protein (E34 TSP) from the E34 TSP gene, then assessed the ability of this bacteriophage protein in the killing of two CBD-resistant strains of Salmonella spp. We also assessed the ability of the tail spike protein to cause bacteria membrane disruption and dehydrogenase depletion. We observed that the combined treatment of CBD-resistant strains of Salmonella with CBD and E34 TSP showed poor killing ability, whereas the mono treatment with E34 TSP showed considerably higher killing efficiency. This study demonstrates that the inhibition of the bacteria by E34 TSP was due in part to membrane disruption and dehydrogenase inactivation by the protein. The results of this work provide an interesting background to highlight the crucial role phage proteins such as E34 TSP could play in pathogenic bacterial control.

Keywords: cannabidiol, resistance, Salmonella, antimicrobials, phages

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Authors: Chuanpeng Zhou

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The effect of dietary carbohydrate (CHO) level on serum physiological response, hepatic antioxidant abilities and heat shock protein 70 (HSP70) expression of Wuchang bream (Megalobrama amblycephala) was studied. Two isonitrogenous (28.56% crude protein) and isolipidic (5.28% crude lipid) diets were formulated to contain 30% or 53% wheat starch. Diets were fed for 90 days to fish in triplicate tanks (28 fish per tank). At the end of feeding trial, significantly higher serum triglyceride level, insulin level, cortisol level, malondialdehyde (MDA) content were observed in fish fed the 53% CHO diet, while significantly lower serum total protein content, alkaline phosphatase (AKP) activity, superoxide dismutase (SOD) activity and total antioxidative capacity (T-AOC) were found in fish fed the 53% CHO diet compared with those fed the 30% diet. The relative level of hepatic heat shock protein 70 mRNA was significantly higher in the 53% CHO group than that in the 30% CHO at 6, 12, and 48 h after feeding. The results of this study indicated that ingestion of 53% dietary CHO impacted the nonspecific immune ability and caused metabolic stress of Megalobrama amblycephala.

Keywords: Megalobrama amblycephala, carbohydrate, fasted and postprandial response, immunity, Hsp70

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Authors: S. El Hasini, M. El Azzouzi, M. De Nobili, K. Azim, A. Zouahri

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Soil salinization is one of the most severe environmental hazards which threaten sustainable agriculture in arid and semi-arid regions, including Morocco. In this regard the application of organic matter to saline soil has confirmed its effectiveness. The present study was aimed to examine the effect of humic acid which represent, among others, the important component of organic matter that contributes to reduce soil salinity. In fact, different composts taken from Agadir (Morocco), with different C/N ratio, were tested. After extraction and purification of humic acid, the interaction with Na2CO3 was carried out. The reduction of salinity is calculated as a value expressed in mg Na2CO3 equivalent/g HA. The results showed that humic acid had generally a significant effect on salinity. In that respect, the hypothesis proposed that carboxylic groups of humic acid create bonds with excess sodium in the soil to form a coherent complex which descends by leaching operation. The comparison between composts was based on C/N ratio, it showed that the compost with the lower ratio C/N had the most important effect on salinity reduction, whereas the compost with higher C/N ratio was less effective. The study is attended also to evaluate the quality of each compost by determining the humification index, we noticed that the compost which have the lowest C/N (20) ratio was relatively less stable, where a greater predominance of the humified substances, when the compost with C/N ratio is 35 exhibited higher stability.

Keywords: compost, humic acid, organic matter, salinity

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Authors: Agnieszka Sobczak-Kupiec, Dagmara Malina, Klaudia Pluta, Wioletta Florkiewicz, Bozena Tyliszczak

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Introduction: Request for compound of phosphorus grows continuously, thus, it is searched for alternative sources of this element. One of these sources could be by-products from meat industry which contain prominent quantity of phosphorus compounds. Hydroxyapatite, which is natural component of animal and human bones, is leading material applied in bone surgery and also in stomatology. This is material, which is biocompatible, bioactive and osteoinductive. Methodology: Hydroxyapatite preparation: As a raw material was applied deproteinized and defatted bone pulp called bone sludge, which was formed as waste in deproteinization process of bones, in which a protein hydrolysate was the main product. Hydroxyapatite was received in calcining process in chamber kiln with electric heating in air atmosphere in two stages. In the first stage, material was calcining in temperature 600°C within 3 hours. In the next stage unified material was calcining in three different temperatures (750°C, 850°C and 950°C) keeping material in maximum temperature within 3.0 hours. Bone sludge: Bone sludge was formed as waste in deproteinization process of bones, in which a protein hydrolysate was the main product. Pork bones coming from the partition of meat were used as a raw material for the production of the protein hydrolysate. After disintegration, a mixture of bone pulp and water with a small amount of lactic acid was boiled at temperature 130-135°C and under pressure4 bar. After 3-3.5 hours boiled-out bones were separated on a sieve, and the solution of protein-fat hydrolysate got into a decanter, where bone sludge was separated from it. Results of the study: The phase composition was analyzed by roentgenographic method. Hydroxyapatite was the only crystalline phase observed in all the calcining products. XRD investigation was shown that crystallization degree of hydroxyapatite was increased with calcining temperature. Conclusion: The researches were shown that phosphorus content is around 12%, whereas, calcium content amounts to 28% on average. The conducted researches on bone-waste calcining at the temperatures of 750-950°C confirmed that thermal utilization of deproteinized bone-waste was possible. X-ray investigations were confirmed that hydroxyapatite is the main component of calcining products, and also XRD investigation was shown that crystallization degree of hydroxyapatite was increased with calcining temperature. Contents of calcium and phosphorus were distinctly increased with calcining temperature, whereas contents of phosphorus soluble in acids were decreased. It could be connected with higher crystallization degree of material received in higher temperatures and its stable structure. Acknowledgements: “The authors would like to thank the The National Centre for Research and Development (Grant no: LIDER//037/481/L-5/13/NCBR/2014) for providing financial support to this project”.

Keywords: bone by-products, bone sludge, calcination, hydroxyapatite

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Authors: A. Ameur Ameur, F. Chougrani, M. Halbouche

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In order to characterize the sheep's milk, we analyzed and compared, in a first stage of our work, the physical and chemical characteristics in two Algerian sheep breeds: Hamra race and race Ouled Djellal breeding at the station the experimental ITELV Ain Hadjar (Saïda Province). Analyses are performed by Ekomilk Ultra-analyzer (EON TRADING LLC, USA), they focused on the pH, density, freezing, fat, total protein, solids-the total dry extract. The results obtained for these parameters showed no significant differences between the two breeds studied. The second stage of this work was the isolation and characterization of milk proteins. For this, we used the precipitation of caseins phi [pH 4.6]. For this, we used the precipitation of caseins Phi (pH 4.6). After extraction, purification and assay, both casein and serum protein fractions were then assayed by the Bradford method and controlled by polyacrylamide gel electrophoresis (PAGE) in the different conditions (native, in the presence of urea and in the presence of SDS). The electrophoretic pattern of milk samples showed the presence similarities of four major caseins variants (αs1-, αs2-β-and k-casein) and two whey proteins (β-lactoglobulin, α-lactalbumin) of two races Hamra and Ouled Djellal. But compared to bovine milk, they have helped to highlight some peculiarities as related to serum proteins (α La β Lg) as caseins, including αs1-Cn.

Keywords: Hamra, Ouled Djellal, protein polymorphism, sheep breeds

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Authors: Hamidreza Saberkari, Mousa Shamsi, Hossein Ahmadi, Saeed Vaali, , MohammadHossein Sedaaghi

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In the recent years, using signal processing tools for accurate identification of the protein coding regions has become a challenge in bioinformatics. Most of the genomic signal processing methods is based on the period-3 characteristics of the nucleoids in DNA strands and consequently, spectral analysis is applied to the numerical sequences of DNA to find the location of periodical components. In this paper, a novel ensemble algorithm for gene selection in DNA sequences has been presented which is based on the combination of Goertzel algorithm and anti-notch filter (ANF). The proposed algorithm has many advantages when compared to other conventional methods. Firstly, it leads to identify the coding protein regions more accurate due to using the Goertzel algorithm which is tuned at the desired frequency. Secondly, faster detection time is achieved. The proposed algorithm is applied on several genes, including genes available in databases BG570 and HMR195 and their results are compared to other methods based on the nucleotide level evaluation criteria. Implementation results show the excellent performance of the proposed algorithm in identifying protein coding regions, specifically in identification of small-scale gene areas.

Keywords: protein coding regions, period-3, anti-notch filter, Goertzel algorithm

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Authors: Hazrat Salman Sidique, Muhammad Tahir Khan, Haq Aman Ullah, Muhammad Mobashar, Muhammad Ishtiaq Sohail Mehmood

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The poor quality conventional feed for the livestock production in Pakistan are wheat straw, tops of sugar cane and tree leaves. To enhance the nutritive value of feed, this study focused on investigating the effects of fibrolytic enzyme (Fibrozyme®, Alltech Inc. Company, USA) at different levels i.e. 0, 5, 10, and 15g/kg of total mix ration on feed intake, digestibility, milk yield and composition, and economics of the ration in Holstein Friesians cows. Twelve Holstein Friesians cows of almost the same age, and lactation stage were randomly allocated into 4 equal groups i.e. A, B, C, and D. Four experimental rations supplemented with Fibrozyme® 0g, 5g, 10g, and 15g/Kg of total mix ration were assigned to these sets correspondingly. The dry matter intake was linearly and significantly (P<0.05) improved. A significant effect of Fibrozyme® was observed for organic matter digestibility, ether extract digestibility, crude fiber digestibility, nitrogen free extract digestibility, and acid detergent fiber digestibility while the results were statistically non-significant for crude protein digestibility, neutral detergent fiber digestibility, and ash digestibility. Milk yield and composition except fat were significantly (P<0.05) increased in all Fibrozyme® treated groups. This study concludes that supplementation of Fibrozyme® at the rate of 15g/Kg total mix ration improved the dry matter intake, nutrients digestibility, and milk production and constituents like protein, lactose, and solid not fat. Therefore, treatment of total mix ration with Fibrozyme® was desirably reasonable and profitable.

Keywords: digestibility, fibrozyme, TMR, digestibility, lactating cow

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Authors: Yamina Boukari, Nashiru Billa, Andrew Morris, Stephen Doughty, Kevin Shakesheff

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Poly (lactic-co-glycolic) acid (PLGA) is a synthetic polymer that can be used in bone tissue engineering with the aim of creating a scaffold in order to support the growth of cells. The formation of microspheres from this polymer is an attractive strategy that would allow for the development of an injectable system, hence avoiding invasive surgical procedures. The aim of this study was to develop a microsphere delivery system for use as an injectable scaffold in bone tissue engineering and evaluate various formulation parameters on its properties. Porous and lysozyme-containing PLGA microspheres were prepared using the double emulsion solvent evaporation method from various molecular weights (MW). Scaffolds were formed by sintering to contain 1 -3mg of lysozyme per gram of scaffold. The mechanical and physical properties of the scaffolds were assessed along with the release of lysozyme, which was used as a model protein. The MW of PLGA was found to have an influence on microsphere size during fabrication, with increased MW leading to an increased microsphere diameter. An inversely proportional relationship was displayed between PLGA MW and mechanical strength of formed scaffolds across loadings for low, intermediate and high MW respectively. Lysozyme release from both microspheres and formed scaffolds showed an initial burst release phase, with both microspheres and scaffolds fabricated using high MW PLGA showing the lowest protein release. Following the initial burst phase, the profiles for each MW followed a similar slow release over 30 days. Overall, the results of this study demonstrate that lysozyme can be successfully incorporated into porous PLGA scaffolds and released over 30 days in vitro, and that varying the MW of the PLGA can be used as a method of altering the physical properties of the resulting scaffolds.

Keywords: bone, microspheres, PLGA, tissue engineering

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Authors: Arash Khorasani Esmaeili, Rosna Mat Taha, Sadegh Mohajer

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Seeds of Trifolium pratense (Red clover) were exposed in vitro for 6 weeks to six levels of lead (Pb) concentrations (0, 50, 100, 150, 200, 250 µM) to analyze the effects on growth, total chlorophyll and total protein contents of grown plants against the lead accumulation. The growth of plants was negatively affected by various levels of lead treatment. The fresh and dry weights, as well as lengths of shoots and roots of grown plants under various lead treatments, were found significantly lower in comparison with the control plants. Total chlorophyll and total soluble protein contents of grown plants under lower concentrations of lead treatment did not show significant differences when compared with the control plants, although they were affected significantly in higher levels of lead accumulation (150-250 µM).

Keywords: trifolium pratense, lead accumulation, chlorophyll content, protein content

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Authors: Min-Shiuan Liou, Yi Shan Yang, Yang-Zhan Huang, Chia-Wen Hsieh

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A high speed growing and butanol-tolerant Clostridium acetobutylicum HOL1 mutant was screened throughout continuous adaption culture with C. acetobutylicum ATCC 824. The HOL1 strain can grow well in 10 g/L butanol contained CGM medium and can produce about 12.8 g /L butanol during 24 hrs. The C. acetobutylicum HOL1 strain was able to produce 166 mM butanol with 21 mM acetone at pH 4.8, resulting in a butanol selectivity (a molar ratio of butanol to total solvents) of 0.79, which is much higher than that (0.6) of the wild-type strain C. acetobutylicum ATCC 824. The acetate and butyrate accumulation were not observed during fermentation of the HOL1 strain. A hyper-butanol producing C. acetobutylicum HOL1 (pBPHS-3), which was created to overexpress the Bacillus psychrosaccharolyticus originated specific heat-shock protein gene, hspX, from a clostridial phosphotransbutyrylase promoter, was studied for its potential to produce a high titer of butanol. Overexpression of hspX resulted in increased final butanol yield 47% and 30% higher than those of the the ATCC824 and the HOL1 strains, respectively. The remarkable high-speed growth and butanol tolerance of strain HOL1 (pBPHS-3) demonstrates that overexpression of heterogeneous stress protein-encoding gene, hspX, could help C. acetobutylicum to effectively produce a high concentration of butanol.

Keywords: Clostridium acetobutylicum, butanol, heat-shock protein, resistance

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Authors: Jessica Pinheiro Silva, Brenda Rabello De Camargo, Daniel Gusmao De Morais, Eliane Ferreira Noronha

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The utilization of lignocellulosic biomass as source of polysaccharides for industrial applications requires an arsenal of enzymes with different mode of action able to hydrolyze its complex and recalcitrant structure. Clostridium thermocellum is gram-positive, thermophilic bacterium producing lignocellulosic hydrolyzing enzymes in the form of multi-enzyme complex, termed celulossomes. This complex has several hydrolytic enzymes attached to a large and enzymically inactive protein known as Cellulosome-integrating protein (CipA), which serves as a scaffolding protein for the complex produced. This attachment occurs through specific interactions between cohesin modules of CipA and dockerin modules in enzymes. The present work aims to construct celulosomes in vitro with the structural protein CipA, a xylanase called Xyn10D and a cellulose called CelJ from C.thermocellum. A mini-scafoldin was constructed from modules derived from CipA containing two cohesion modules. This was cloned and expressed in Escherichia coli. The other two genes were cloned under the control of the alcohol oxidase 1 promoter (AOX1) in the vector pPIC9 and integrated into the genome of the methylotrophic yeast Pichia pastoris GS115. Purification of each protein is being carried out. Further studies regarding enzymatic activity of the cellulosome is going to be evaluated. The cellulosome built in vitro and composed of mini-CipA, CelJ and Xyn10D, can be very interesting for application in industrial processes involving the degradation of plant biomass.

Keywords: cellulosome, CipA, Clostridium thermocellum, cohesin, dockerin, yeast

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Authors: Bello Muhammad Dogon Kade

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The study was conducted to determine the effect of a treated groundnut (Arachis hypogaea) shell in a complete diet on blood metabolites and microbial load in fattening Yankasa rams. The study was conducted at the Teaching and Research Farm (Small Ruminants Unit of Animal Science Department, Faculty of Agriculture, Ahmadu Bello University, Zaria. Each kilogram of groundnut shell was treated with 5% urea and 5% lime for treatments 2 (UTGNS) and 3 (LTGNS), respectively. For treatment 4 (ULTGNS), 1 kg of groundnut shell was treated with 2.5% urea and 2.5% lime, but the shell in treatment 1 was not treated (UNTGNS). Sixteen Yankasa rams were used and randomly assigned to the four treatment diets with four animals per treatment in a completely randomized design (CRD). The diet was formulated to have 14% crude protein (CP) content. Rumen fluid was collected from each ram at the end of the experiment at 0 and 4 hours post-feeding. The samples were then put in a 30 ml bottle and acidified with 5 drops of concentrated sulphuric (0.1N H₂SO4) acid to trap ammonia. The results of the blood metabolites showed that the mean values of NH₃-N differed significantly (P<0.05) among the treatment groups, with rams in the ULTGNS diet having the highest significant value (31.96 mg/L). TVFs were significantly (P<0.05) higher in rams fed UNTGNS diet and higher in total nitrogen; the effect of sampling periods revealed that NH3N, TVFs and TP were significantly (P<0.05) higher in rumen fluid collected 4hrs post feeding among the rams across the treatment groups, but rumen fluid pH was significantly (p<0.05) higher in 0-hour post-feeding in all the rams in the treatment diets. In the treatment and sampling period’s interaction effects, animals on the ULTGNS diet had the highest mean values of NH3N in both 0 and 4 hours post-feeding and were significantly (P<0.5) higher compared to rams on the other treatment diets. Rams on the UTGNS diet had the highest bacteria load of 4.96X105/ml, which was significantly (P<0.05) higher than a microbial load of animals fed UNTGNS, LTGNS and ULTGNS diets. However, protozoa counts were significantly (P<0.05) higher in rams fed the UTGNS diet than those followed by the ULTGNS diet. The results showed that there was no significant difference (P>0.05) in the bacteria count of the animals at both 0 and 4 hours post-feeding. But rumen fungi and protozoa load at 0 hours were significantly (P<0.05) higher than at 4 hours post-feeding. The use of untreated ground groundnut shells in the diet of fattening Yankasa ram is therefore recommended.

Keywords: blood metabolites, microbial load, volatile fatty acid, ammonia, total protein

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Authors: Mohamed A. Morsy, Azza A. K. El-Sheikh, Abdulla Y. Al-Taher

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The aim of the present study is to investigate the effect of resveratrol (RES) on methotrexate (MTX)-induced testicular damage. RES (10 mg/kg/day) was given for 8 days orally and MTX (20 mg/kg i.p.) was given at day 4 of experiment, with or without RES in rats. MTX decreased serum testosterone, induced histopathological testicular damage, increased testicular tumor necrosis factor-α level and expression of nuclear factor-κB and cyclooxygenase-2. In MTX/RES group, significant reversal of these parameters was noticed, compared to MTX group. Testicular expression of multidrug resistance protein (Mrp) 3 was three- and five-folds higher in RES- and MTX/RES-treated groups, respectively. In vitro, using prostate cancer cells, each of MTX and RES alone induced cytotoxicity with IC50 0.18 ± 0.08 and 20.5 ± 3.6 µM, respectively. RES also significantly enhanced cytotoxicity of MTX. In conclusion, RES appears to have dual beneficial effect, as it promotes MTX tumor cytotoxicity, while protecting the testes, probably via up-regulation of testicular Mrp3 as a novel mechanism.

Keywords: resveratrol, methotrexate, multidrug resistance protein 3, tumor necrosis factor-α, nuclear factor-κB, cyclooxygenase-2

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Authors: Mohamed M. Ibrahim, Gaber A. M. Mersal, Salih Al-Juaid, Samir A. El-Shazly

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A series of mixed ligand complexes, viz., [Cu(BPy)(Gly)X]Y {X = Cl (1), Y = 0; X = 0, Y = ClO4- (2); X = H2O, Y = NO3- (3); X = H2O, Y = CH3COO- (4); and [Cu(BPy)(Gly)-(H2O)]2(SO4) (5) have been synthesized. Their structures and properties were characterized by elemental analysis, thermal analaysis, IR, UV–vis, and ESR spectroscopy, as well as electrochemical measurements including cyclic voltammetry, electrical molar conductivity, and magnetic moment measurements. Complexes 1 and 2 formed slightly distorted square-pyramidal coordination geometries of CuN3OCl and CuN3O2, respectively in which the N,O-donor glycine and N,N-donor bipyridyl bind at the basal plane with chloride ion or water as the axial ligand. Complex 3 shows square planar CuN3O coordination geometry, which exhibits chemically significant hydrogen bonding interactions besides showing coordination polymer formation. The superoxide dismutase and catalase-like activities of all complexes were tested and were found to be promising candidates as durable electron-transfer catalyst being close to the efficiency of the mimicking enzymes displaying either catalase or tyrosinase activity to serve for complete reactive oxygen species (ROS) detoxification, both with respect to superoxide radicals and related peroxides. The DNA binding interaction with super coiled pGEM-T plasmid DNA was investigated by using spectral (absorption and emission) titration and electrochemical techniques. The results revealed that DNA intercalate with complexes 1 and 2 through the groove binding mode. The calculated intrinsic binding constant (Kb) of 1 and 2 were 4.71 and 2.429 × 105 M−1, respectively. Gel electrophoresis study reveals the fact that both complexes cleave super coiled pGEM-T plasmid DNA to nicked and linear forms in the absence of any additives. On the other hand, the interaction of both complexes with DNA, the quasi-reversible CuII/CuI redox couple slightly improves its reversibility with considerable decrease in current intensity. All the experimental results indicate that the bipyridyl mixed copper(II) complex (1) intercalate more effectively into the DNA base pairs.

Keywords: enzyme mimics, mixed ligand complexes, X-ray structures, antioxidant, DNA-binding, DNA cleavage

Procedia PDF Downloads 542

Authors: Xinyu Wei, Mingming Peng, Kenji Kamiya, Eika Qian

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Iso-stearic acid as a saturated fatty acid with a branched chain shows a low pour point, high oxidative stability and great biodegradability. The industrial production of iso-stearic acid involves first isomerizing unsaturated fatty acids into branched-chain unsaturated fatty acids (BUFAs), followed by hydrogenating the branched-chain unsaturated fatty acids to obtain iso-stearic acid. However, the production yield of iso-stearic acid is reportedly less than 30%. In recent decades, extensive research has been conducted on branched fatty acids. Most research has replaced acidic clays with zeolites due to their high selectivity, good thermal stability, and renewability. It was reported that isomerization of unsaturated fatty acid occurred mainly inside the zeolite channel. In contrast, the production of by-products like dimer acid mainly occurs at acid sites outside the surface of zeolite. Further, the deactivation of catalysts is attributed to the pore blockage of zeolite. In the present study, micro/mesoporous ZSM-22 zeolites were developed. It is clear that the synthesis of a micro/mesoporous ZSM-22 zeolite is regarded as the ideal strategy owing to its ability to minimize coke formation. Different mesoporosities micro/mesoporous H-ZSM-22 zeolites were prepared through recrystallization of ZSM-22 using sodium hydroxide solution (0.2-1M) with cetyltrimethylammonium bromide template (CTAB). The structure, morphology, porosity, acidity, and isomerization performance of the prepared catalysts were characterized and evaluated. The dissolution and recrystallization process of the H-ZSM-22 microporous zeolite led to the formation of approximately 4 nm-sized mesoporous channels on the outer surface of the microporous zeolite, resulting in a micro/mesoporous material. This process increased the weak Brønsted acid sites at the pore mouth while reducing the total number of acid sites in ZSM-22. Finally, an activity test was conducted using oleic acid as a model compound in a fixed-bed reactor. The activity test results revealed that micro/mesoporous H-ZSM-22 zeolites exhibited a high isomerization activity, reaching >70% selectivity and >50% yield of BUFAs. Furthermore, the yield of oligomers was limited to less than 20%. This demonstrates that the presence of mesopores in ZSM-22 enhances contact between the feedstock and the active sites within the catalyst, thereby increasing catalyst activity. Additionally, a portion of the dissolved and recrystallized silica adhered to the catalyst's surface, covering the surface-active sites, which reduced the formation of oligomers. This study offers distinct insights into the production of iso-stearic acid using a fixed-bed reactor, paving the way for future research in this area.

Keywords: Iso-stearic acid, oleic acid, skeletal isomerization, micro/mesoporous, ZSM-22

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Authors: Nusrat Praween, Krishna Thej Pammi Guru, Palash Kumar Basu

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Designing non-invasive biosensors for cancer diagnosis is essential for developing an affordable and specific tool to measure cancer-related exosome biomarkers. Exosomes, released by healthy as well as cancer cells, contain valuable information about the biomarkers of various diseases, including cancer. Despite the availability of various isolation techniques, ultracentrifugation is the standard technique that is being employed. Post isolation, exosomes are traditionally exposed to detergents for extracting their proteins, which can often lead to protein degradation. Further to this, it is very essential to develop a sensing platform for the quantification of clinically relevant proteins in a wider range to ensure practicality. In this study, exosomes were immobilized on the Au Screen Printed Electrode (SPE) using EDC/NHS chemistry to facilitate binding. After immobilizing the exosomes on the screen-printed electrode (SPE), we investigated the impact of the electric field by applying various voltages to induce exosome lysis and release their contents. The lysed solution was used for sensing TSG101, a crucial biomarker associated with various cancers, using both faradaic and non-faradaic electrochemical impedance spectroscopy (EIS) methods. The results of non-faradaic and faradaic EIS were comparable and showed good consistency, indicating that non-faradaic sensing can be a reliable alternative. Hence, the non-faradaic sensing technique was used for label-free quantification of the TSG101 biomarker. The results were validated using ELISA. Our electrochemical immunosensor demonstrated a consistent response of TSG101 from 125 pg/mL to 8000 pg/mL, with a detection limit of 0.125 pg/mL at room temperature. Additionally, since non-faradic sensing is label-free, the ease of usage and cost of the final sensor developed can be reduced. The proposed immunosensor is capable of detecting the TSG101 protein at low levels in healthy serum with good sensitivity and specificity, making it a promising platform for biomarker detection.

Keywords: biosensor, exosomes isolation on SPE, electric field lysis of exosome, EIS sensing of TSG101

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Authors: Rasha A.Ibrahim El-Ghazawy, Ashraf M. El-Saeed, Heusin El-Shafey, M. Abdel-Raheim, Maher A. El-Sockary

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Two thermotropic liquid crystalline curing agents based on abietic acid with different mesogens (LCC1 and LCC2) were synthesized for producing thermally stable liquid crystal networks suitable for high performance epoxy coatings. Differential scanning calorimetry (DSC) and polarized optical microscope (POM) was used to identify the liquid crystal phase transformation temperatures and texture, respectively. POM micro graphs for both LCCs revealing cholesteric texture. A multifunctional epoxy resin with two abietic acid moieties was also synthesized. Dynamic mechanical (DMA) and thermogravimetric (TGA) analyses show that the fully bio-based cured epoxies by either LCCs possess high glass transition temperature (Tg), high modulus (G`) and improved thermal stability. The chemical structure of the synthesized LCCs and epoxy resin was investigated through FTIR and 1HNMR spectroscopic techniques.

Keywords: abietic acid, dynamic mechanical analysis, epoxy resin, liquid crystal, thermo gravimetric analysis

Procedia PDF Downloads 359