Search results for: whey protein isolated

Authors: D. Dixon, J. Nicol, J. A. Coulter, E. Harrison

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The ease with which they can be functionalized, combined with their excellent biocompatibility, make gold nanoparticles (AuNPs) ideal candidates for various applications in nanomedicine. Indeed several promising treatments are currently undergoing human clinical trials (CYT-6091 and Auroshell). A successful nanoparticle treatment must first evade the immune system, then accumulate within the target tissue, before enter the diseased cells and delivering the payload. In order to create a clinically relevant drug delivery system, contrast agent or radiosensitizer, it is generally necessary to functionalize the AuNP surface with multiple groups; e.g. Polyethylene Glycol (PEG) for enhanced stability, targeting groups such as antibodies, peptides for enhanced internalization, and therapeutic agents. Creating and characterizing the biological response of such complex systems remains a challenge. The two commonly used methods to attach multiple groups to the surface of AuNPs are the creation of a mixed monolayer, or by binding groups to the AuNP surface using a bi-functional PEG linker. While some excellent in-vitro and animal results have been reported for both approaches further work is necessary to directly compare the two methods. In this study AuNPs capped with both PEG and a Receptor Mediated Endocytosis (RME) peptide were prepared using both mixed monolayer and PEG linker approaches. The PEG linker used was SH-PEG-SGA which has a thiol at one end for AuNP attachment, and an NHS ester at the other to bind to the peptide. The work builds upon previous studies carried out at the University of Ulster which have investigated AuNP synthesis, the influence of PEG on stability in a range of media and investigated intracellular payload release. 18-19nm citrate capped AuNPs were prepared using the Turkevich method via the sodium citrate reduction of boiling 0.01wt% Chloroauric acid. To produce PEG capped AuNPs, the required amount of PEG-SH (5000Mw) or SH-PEG-SGA (3000Mw Jenkem Technologies) was added, and the solution stirred overnight at room temperature. The RME (sequence: CKKKKKKSEDEYPYVPN, Biomatik) co-functionalised samples were prepared by adding the required amount of peptide to the PEG capped samples and stirring overnight. The appropriate amounts of PEG-SH and RME peptide were added to the AuNP to produce a mixed monolayer consisting of approximately 50% PEG and 50% RME. The PEG linker samples were first fully capped with bi-functional PEG before being capped with RME peptide. An increase in diameter from 18-19mm for the ‘as synthesized’ AuNPs to 40-42nm after PEG capping was observed via DLS. The presence of PEG and RME peptide on both the mixed monolayer and PEG linker co-functionalized samples was confirmed by both FTIR and TGA. Bi-functional PEG linkers allow the entire AuNP surface to be capped with PEG, enabling in-vitro stability to be achieved using a lower molecular weight PEG. The approach also allows the entire outer surface to be coated with peptide or other biologically active groups, whilst also offering the promise of enhanced biological availability. The effect of mixed monolayer versus PEG linker attachment on both stability and non-specific protein corona interactions was also studied.

Keywords: nanomedicine, gold nanoparticles, PEG, biocompatibility

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Authors: Ashish Thakur

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This technical paper was written in view of focusing the biomaterials and its various applications in modern industries. Author tires to elaborate not only the medical, infect plenty of application in other industries. The scope of the research area covers the wide range of physical, biological and chemical sciences that underpin the design of biomaterials and the clinical disciplines in which they are used. A biomaterial is now defined as a substance that has been engineered to take a form which, alone or as part of a complex system, is used to direct, by control of interactions with components of living systems, the course of any therapeutic or diagnostic procedure. Biomaterials are invariably in contact with living tissues. Thus, interactions between the surface of a synthetic material and biological environment must be well understood. This paper reviews the benefits and challenges associated with surface modification of the metals in biomedical applications. The paper also elaborates how the surface characteristics of metallic biomaterials, such as surface chemistry, topography, surface charge, and wettability, influence the protein adsorption and subsequent cell behavior in terms of adhesion, proliferation, and differentiation at the biomaterial–tissue interface. The chapter also highlights various techniques required for surface modification and coating of metallic biomaterials, including physicochemical and biochemical surface treatments and calcium phosphate and oxide coatings. In this review, the attention is focused on the biomaterial-associated infections, from which the need for anti-infective biomaterials originates. Biomaterial-associated infections differ markedly for epidemiology, aetiology and severity, depending mainly on the anatomic site, on the time of biomaterial application, and on the depth of the tissues harbouring the prosthesis. Here, the diversity and complexity of the different scenarios where medical devices are currently utilised are explored, providing an overview of the emblematic applicative fields and of the requirements for anti-infective biomaterials. In addition to this, chapter introduces nanomedicine and the use of both natural and synthetic polymeric biomaterials, focuses on specific current polymeric nanomedicine applications and research, and concludes with the challenges of nanomedicine research. Infection is currently regarded as the most severe and devastating complication associated to the use of biomaterials. Osteoporosis is a worldwide disease with a very high prevalence in humans older than 50. The main clinical consequences are bone fractures, which often lead to patient disability or even death. A number of commercial biomaterials are currently used to treat osteoporotic bone fractures, but most of these have not been specifically designed for that purpose. Many drug- or cell-loaded biomaterials have been proposed in research laboratories, but very few have received approval for commercial use. Polymeric nanomaterial-based therapeutics plays a key role in the field of medicine in treatment areas such as drug delivery, tissue engineering, cancer, diabetes, and neurodegenerative diseases. Advantages in the use of polymers over other materials for nanomedicine include increased functionality, design flexibility, improved processability, and, in some cases, biocompatibility.

Keywords: nanomedicine, tissue, infections, biomaterials

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Authors: Swati Mishra

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In recent years, there has been an explosion in Gold NanoParticle (GNP) research, with a rapid increase in publications in diverse fields, including imaging, bioengineering, and molecular biology. GNPs exhibit unique physicochemical properties, including surface plasmon resonance (SPR) and bind amine and thiol groups, allowing surface modification and use in biomedical applications. Nanoparticle functionalization is the subject of intense research at present, with rapid progress being made towards developing biocompatible, multi-functional particles. In the present study, the photochemical method has been done to functionalize various-shaped GNPs like nanostars by the molecules like ninhydrin. Ninhydrin is bactericidal, virucidal, fungicidal, antigen-antibody reactive, and used in fingerprint technology in forensics. The GNPs functionalized with ninhydrin efficiently will bind to the amino acids on the target protein, which is of eminent importance during the pandemic, especially where long-term treatments of COVID- 19 bring many side effects of the drugs. The photochemical method is adopted as it provides low thermal load, selective reactivity, selective activation, and controlled radiation in time, space, and energy. The GNPs exhibit their characteristic spectrum, but a distinctly blue or redshift in the peak will be observed after UV irradiation, ensuring efficient ninhydrin binding. Now, the bound ninhydrin in the GNP carrier, upon chemically reacting with any amino acid, will lead to the formation of Rhumann purple. A common method of GNP production includes citrate reduction of Au [III] derivatives such as aurochloric acid (HAuCl4) in water to Au [0] through a one-step synthesis of size-tunable GNPs. The following reagents are prepared to validate the approach. Reagent A solution 1 is0.0175 grams ninhydrin in 5 ml Millipore water Reagent B 30 µl of HAuCl₄.3H₂O in 3 ml of solution 1 Reagent C 1 µl of gold nanostars in 3 ml of solution 1 Reagent D 6 µl of cetrimonium bromide (CTAB) in 3 ml of solution1 ReagentE 1 µl of gold nanostars in 3 ml of ethanol ReagentF 30 µl of HAuCl₄.₃H₂O in 3 ml of ethanol ReagentG 30 µl of HAuCl₄.₃H₂O in 3 ml of solution 2 ReagentH solution 2 is0.0087 grams ninhydrin in 5 ml Millipore water ReagentI 30 µl of HAuCl₄.₃H₂O in 3 ml of water The reagents were irradiated at 254 nm for 15 minutes, followed by their UV Visible spectroscopy. The wavelength was selected based on the one reported for excitation of a similar molecule Pthalimide. It was observed that the solution B and G deviate around 600 nm, while C peaks distinctively at 567.25 nm and 983.9 nm. Though it is tough to say about the chemical reaction happening, butATR-FTIR of reagents will ensure that ninhydrin is not forming Rhumann purple in the absence of amino acids. Therefore, these experiments, we achieved the functionalization of gold nanostars with ninhydrin corroborated by the deviation in the spectrum obtained in a mixture of GNPs and ninhydrin irradiated with UV light. It prepares them as a carrier molecule totake up amino acids for targeted delivery or germicidal action.

Keywords: gold nanostars, ninhydrin, photochemical method, UV visible specgtroscopy

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Authors: Ripon Sarkar, Kabita Chaterjee, Ananya Barui

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Smoking is one of the leading causes of oral cancer. Cigarette smoke affects various cellular parameters and alters molecular metabolism of cells. Epithelial cells losses their cytoskeleton structure, membrane integrity, cellular polarity that subsequently initiates the process of epithelial cells to mesenchymal transition due to long exposure of cigarette smoking. It changes the normal cellular metabolic activity which induces oxidative stress and enhances the reactive oxygen spices (ROS) formation. Excessive ROS and associated oxidative stress are considered to be a driving force in alteration in cellular phenotypes, polarity distribution and mitochondrial metabolism. Noninvasive assessment of such parameters plays essential role in development of routine screening system for early diagnosis of oral cancer. Electrical cell-substrate impedance sensing (ECIS) is one of such method applied for detection of cellular membrane impedance which can be correlated to cell membrane integrity. Present study intends to explore the alteration in cellular impedance along with the expression of cellular polarity molecules and cytoskeleton distributions in oral epithelial cells of habitual smokers and to correlate the outcome to that of clinically diagnosed oral leukoplakia and oral squamous cell carcinoma patients. Total 80 subjects were categorized into four study groups: nonsmoker (NS), cigarette smoker (CS), oral leukoplakia (OLPK) and oral squamous cell carcinoma (OSCC). Cytoskeleton distribution was analyzed by staining of actin filament and generation of ROS was measured using assay kit using standard protocol. Cell impedance was measured through ECIS method at different frequencies. Expression of E-cadherin and protease-activated receptor (PAR) proteins were observed through immune-fluorescence method. Distribution of actin filament is well organized in NS group however; distribution pattern was grossly varied in CS, OLPK and OSCC. Generation of ROS was low in NS which subsequently increased towards OSCC. Expressions of E-cadherin and change in cellular electrical impedance in different study groups indicated the hallmark of cancer progression from NS to OSCC. Expressions of E-cadherin, PAR protein, and cell impedance were decreased from NS to CS and farther OSCC. Generally, the oral epithelial cells exhibit apico-basal polarity however with cancer progression these cells lose their characteristic polarity distribution. In this study expression of polarity molecule and ECIS observation indicates such altered pattern of polarity among smoker group. Overall the present study monitored the alterations in intracellular ROS generation and cell metabolic function, membrane integrity in oral epithelial cells in cigarette smokers. Present study thus has clinical significance, and it may help in developing a noninvasive technique for early diagnosis of oral cancer amongst susceptible individuals.

Keywords: cigarette smoking, early oral cancer detection, electric cell-substrate impedance sensing, noninvasive screening

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Authors: José Silvestre Mendoza Figueroa, Anders Kvarnheden, Jesús Méndez Lozano, Edgar Antonio Rodríguez Negrete, Manuel Soriano García

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Agroindustrial plants such as cereals and pseudo cereals offer a substantial source of biomacromolecules, as they contain large amounts per tissue-gram of proteins, polysaccharides and lipids in comparison with other plants. In particular, Amaranthus hypochondriacus seeds have high levels of proteins in comparison with other cereal and pseudo cereal species, which makes the plant a good source of bioactive molecules such as peptides. Geminiviruses are one principal class of pathogens that causes important economic losses in crops, affecting directly the development and production of the plant. One such virus is the Tomato yellow leaf curl virus (TYLCV), which affects mainly Solanacea family plants such as tomato species. The symptoms of the disease are curling of leaves, chlorosis, dwarfing and floral abortion. The aim of this work was to get peptides derived from enzymatic hydrolysis of globulins and albumins from amaranth seeds with specific recognition of the replication origin in the TYLCV genome, and to test the antiviral activity on host plants with the idea to generate a direct control of this viral infection. Globulins and albumins from amaranth were extracted, the fraction was enzymatically digested with papain, and the aromatic peptides fraction was selected for further purification. Six peptides were tested against the replication origin (OR) using affinity assays, surface resonance plasmon and fluorescent titration, and two of these peptides showed high affinity values to the replication origin of the virus, dissociation constant values were calculated and showed specific interaction between the peptide Ampep1 and the OR. An in vitro replication test of the total TYLCV DNA was performed, in which the peptide AmPep1 was added in different concentrations to the system reaction, which resulted in a decrease of viral DNA synthesis when the peptide concentration increased. Also, we showed that the peptide can decrease the complementary DNA chain of the virus in Nicotiana benthamiana leaves, confirming that the peptide binds to the OR and that its expected mechanism of action is to decrease the replication rate of the viral genome. In an infection assay, N. benthamiana plants were agroinfected with TYLCV-Israel and TYLCV-Guasave. After confirming systemic infection, the peptide was infiltrated in new infected leaves, and the plants treated with the peptide showed a decrease of virus symptoms and viral titer. In order to confirm the antiviral activity in a commercial crop, tomato plants were infected with TYLCV. After confirming systemic infection, plants were infiltrated with peptide solution as above, and the symptom development was monitored 21 days after treatment, showing that tomato plants treated with peptides had lower symptom rates and viral titer. The peptide was also tested against other begomovirus such as Pepper huasteco yellow vein virus (PHYVV-Guasave), showing a decrease of symptoms in N. benthamiana infected plants. The model of direct biochemical control of TYLCV infection shown in this work can be extrapolated to other begomovirus infections, and the methods reported here can be used for design of antiviral agrochemicals for other plant virus infections.

Keywords: agrochemical screening, antiviral, begomovirus, geminivirus, peptides, plasmon, TYLCV

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Authors: Amer Ait Sidhoum

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Interest in sustainability has gained ground among practitioners, academics and policy-makers due to growing stakeholders’ awareness of environmental and social concerns. This is particularly true for agriculture. However, relatively little research has been conducted on the quantification of social sustainability and the contribution of social issues to the agricultural production efficiency. This research's main objective is to propose a method for evaluating prices of social outputs, more precisely shadow prices, by allowing for the stochastic nature of agricultural production that is to say for production uncertainty. In this article, the assessment of social outputs’ shadow prices is conducted within the methodological framework of nonparametric Data Envelopment Analysis (DEA). An output-oriented directional distance function (DDF) is implemented to represent the technology of a sample of Catalan arable crop farms and derive the efficiency scores the overall production technology of our sample is assumed to be the intersection of two different sub-technologies. The first sub-technology models the production of random desirable agricultural outputs, while the second sub-technology reflects the social outcomes from agricultural activities. Once a nonparametric production technology has been represented, the DDF primal approach can be used for efficiency measurement, while shadow prices are drawn from the dual representation of the DDF. Computing shadow prices is a method to assign an economic value to non-marketed social outcomes. Our research uses cross sectional, farm-level data collected in 2015 from a sample of 180 Catalan arable crop farms specialized in the production of cereals, oilseeds and protein (COP) crops. Our results suggest that our sample farms show high performance scores, from 85% for the bad state of nature to 88% for the normal and ideal crop growing conditions. This suggests that farm performance is increasing with an improvement in crop growth conditions. Results also show that average shadow prices of desirable state-contingent output and social outcomes for efficient and inefficient farms are positive, suggesting that the production of desirable marketable outputs and of non-marketable outputs makes a positive contribution to the farm production efficiency. Results also indicate that social outputs’ shadow prices are contingent upon the growing conditions. The shadow prices follow an upward trend as crop-growing conditions improve. This finding suggests that these efficient farms prefer to allocate more resources in the production of desirable outputs than of social outcomes. To our knowledge, this study represents the first attempt to compute shadow prices of social outcomes while accounting for the stochastic nature of the production technology. Our findings suggest that the decision-making process of the efficient farms in dealing with social issues are stochastic and strongly dependent on the growth conditions. This implies that policy-makers should adjust their instruments according to the stochastic environmental conditions. An optimal redistribution of rural development support, by increasing the public payment with the improvement in crop growth conditions, would likely enhance the effectiveness of public policies.

Keywords: data envelopment analysis, shadow prices, social sustainability, sustainable farming

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Authors: Aleksandra Rajewska-Rager, Maria Skibinska, Monika Dmitrzak-Weglarz, Natalia Lepczynska, Pawel Kapelski, Joanna Pawlak, Joanna Hauser

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Introduction: Inflammation and cytokines have emerged as a promising target in mood disorders research; however there are still very limited numbers of study regarding inflammatory alterations among adolescents and young adults with mood disorders. The Macrophage Migration Inhibitory Factor (MIF) and Stem Cell Factor (SCF) are the pleiotropic cytokines which may play an important role in mood disorders pathophysiology. The aim of this study was to investigate levels of these factors in serum of adolescent and young adults with mood disorders compared to healthy controls. Subjects: We involved 79 patients aged 12-24 years in 2-year follow-up study with a primary diagnosis of mood disorders: bipolar disorder (BP) and unipolar disorder with BP spectrum. Study group includes 23 males (mean age 19.08, SD 3.3) and 56 females (18.39, SD 3.28). Control group consisted 35 persons: 7 males (20.43, SD 4.23) and 28 females (21.25, SD 2.11). Clinical diagnoses according to DSM-IV-TR criteria were assessed using Kiddie-Schedule for Affective Disorders and Schizophrenia-Present and Lifetime Version (K-SADS-PL) and Structured Clinical Interview for the Diagnostic and Statistical Manual (SCID) in young adults respectively. Clinical assessment includes evaluation of clinical factors and symptoms severity (rated using the Hamilton Depression Rating Scale and Young Mania Rating Scale). Clinical and biological evaluations were made at control visits respectively at baseline (week 0), euthymia (at month 3 or 6) and after 12 and 24 months. Methods: Serum protein concentration was determined by Enzyme-Linked Immunosorbent Assays (ELISA) method. Human MIF and SCF DuoSet ELISA kits were used. In the analyses non-parametric tests were used: Mann-Whitney U test, Kruskal-Wallis ANOVA, Friedman’s ANOVA, Wilcoxon signed rank test, Spearman correlation. We defined statistical significance as p < 0.05. Results: Comparing MIF and SCF levels between acute episode of depression/hypo/mania at baseline and euthymia (at month 3 or 6) we did not find any statistical differences. At baseline patients with age above 18 years old had decreased MIF level compared to patients younger than 18 years. MIF level at baseline positively correlated with age (p=0.004). Positive correlations of SCF level at month 3 and 6 with depression or mania occurrence at month 24 (p=0.03 and p=0.04, respectively) was detected. Strong correlations between MIF and SCF levels at baseline (p=0.0005) and month 3 (p=0.03) were observed. Discussion: Our results did not show any differences in MIF and SCF levels between acute episode of depression/hypo/mania and euthymia in young patients. Further studies on larger groups are recommended. Grant was founded by National Science Center in Poland no 2011/03/D/NZ5/06146.

Keywords: cytokines, MIF, mood disorders, SCF

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Authors: Natalia V. Naryzhnaya, Alexandr V. Mukhomedzyanov, Ivan A. Derkachev, Boris K. Kurbatov, Leonid N. Maslov

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Background: Techniques of adaptation to hypoxia and remote postconditioning (RPost) have great prospects for use in the clinic. However, recent studies have shown low efficacy of remote postconditioning in patients with AMI. We hypothesize that the reasons for this inefficiency may be metabolic disorders, which are very common, especially in patients with cardiovascular disease, and age of patients. The purpose of the study was to reveal the effectiveness of adaptation to chronic hypoxia and RPost. To determine the possible relationship between the decrease in the effectiveness of projective impacts and disorders of carbohydrate and lipid metabolism. Design: The study was carried out on Wistar rats 60 day old. MetS was induced by high-carbohydrate, high-fat diet (HСHFD). Modeling MS led to the formation of obesity, hypertension, impaired lipid and carbohydrate metabolism, hyperleptinemia, and moderate stress. Groups with adaptation to chronic hypoxia were subjected to hypoxia for 21 days at 12% O2 and 0.3% CO2 after complete of HСHFD. All animals were subjected to 45 min coronary occlusion and 120 min reperfusion. Groups with RPost, immediately after the end of ischemia, tourniquets were applied to the hind limbs in the area of the hip joint (3 times in the mode of 5 min ischemia, 5 min reperfusion). Results: RPost led to a twofold reduction of infarct size in rats with intact metabolism (р < 0.0001), while in rats with MetS, a decrease in infarct size during RPost was 25 % (p = 0.00003). A direct correlation was found between of infarct size during RPost and the serum leptin level of rats with MetC (r = 0.85). The presented data suggested that a decrease in the efficiency of remote postconditioning in rats with diet-induced metabolic syndrome depends on serum leptin. Chronic hypoxia resulted in a 38% reduced in infarct size in metabolically intact rats. The decrease of cardioprotection was observed in rats with chronic hypoxia and MetS. Infarct size showed a direct correlation with impaired glucose tolerance (AUC, glucose tolerance test, r = 0.034) and serum triglyceride levels (r = 0.39). Our study showed the dependence of cardioprotection in rats with metabolic syndrome during chronic hypoxia and DPost on opioids in the blood serum and myocardium, protein kinase C and NO synthase activity. Conclusion: The results obtained showed that the infarct-limiting efficiency of adaptation to hypoxia and remote postconditioning is reduced or completely absent in animals with metabolic syndrome. The increase in the infarction, in this case, directly depends on the disturbances in carbohydrate. lipid metabolism and opioids signaling. Funding: Investigation of effectiveness of chronic hypoxia at the metabolic syndrome was carried out within the support of Russian Science Foundation Grant 22-15-00048. Studies of the mechanisms of arterial hypertension in induced metabolic syndrome were carried out within the framework of the state assignment (122020300042-4). The work was performed using the Center for Collective Use "Medical Genomics".

Keywords: chronic hypoxia, opioids, remote postconditioning, metabolic syndrome

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Authors: Francisco Pereira, António Augusto Vicente, Filipe Vaz, Joel Borges, Pedro Geada

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Light is a growth-limiting factor in microalgae cultivation, where factors like spectral components, intensity, and duration, often characterized by its wavelength, are well-reported to have a substantial impact on cell growth rates and, consequently, photosynthetic performance and mitigation of CO2, one of the most significant greenhouse gases (GHGs). Photobioreactors (PBRs) are commonly used to grow microalgae under controlled conditions, but they often fail to provide an even light distribution to the cultures. For this reason, there is a pressing need for innovations aiming at enhancing the efficient utilization of light. So, one potential approach to address this issue is by implementing plasmonic films, such as the localized surface plasmon resonance (LSPR). LSPR is an optical phenomenon connected to the interaction of light with metallic nanostructures. LSPR excitation is characterized by the oscillation of unbound conduction electrons of the nanoparticles coupled with the electromagnetic field from incident light. As a result of this excitation, highly energetic electrons and a strong electromagnetic field are generated. These effects lead to an amplification of light scattering, absorption, and extinction of specific wavelengths, contingent on the nature of the employed nanoparticle. Thus, microalgae might benefit from this biotechnology as it enables the selective filtration of inhibitory wavelengths and harnesses the electromagnetic fields produced, which could lead to enhancements in both biomass and metabolite productivity. This study aimed at implementing and evaluating a “plasmon-enhanced PBR”. The goal was to utilize LSPR thin films to enhance the growth and CO2 bio-fixation rate of Chlorella vulgaris. The internal/external walls of the PBRs were coated with a TiO2 matrix containing different nanoparticles (Au, Ag, and Au-Ag) in order to evaluate the impact of this approach on microalgae’s performance. Plasmonic films with distinct compositions resulted in different Chlorella vulgaris growth, ranging from 4.85 to 6.13 g.L-1. The highest cell concentrations were obtained with the metallic Ag films, demonstrating a 14% increase compared to the control condition. Moreover, it appeared to be no differences in growth between PBRs with inner and outer wall coatings. In terms of CO2 bio-fixation, distinct rates were obtained depending on the coating applied, ranging from 0.42 to 0.53 gCO2L-1d-1. Ag coating was demonstrated to be the most effective condition for carbon fixation by C. vulgaris. The impact of LSPR films on the biochemical characteristics of biomass (e.g., proteins, lipids, pigments) was analysed as well. Interestingly, Au coating yielded the most significant enhancements in protein content and total pigments, with increments of 15 % and 173 %, respectively, when compared to the PBR without any coating (control condition). Overall, the incorporation of plasmonic films in PBRs seems to have the potential to improve the performance and efficiency of microalgae cultivation, thereby representing an interesting approach to increase both biomass production and GHGs bio-mitigation.

Keywords: CO₂ bio-fixation, plasmonic effect, photobioreactor, photosynthetic microalgae

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Authors: Rebeca P Ramos-Bueno, Maria Jose Gonzalez-Fernandez, Rosa M. Moreno-Zamora, Antonia Barros Heras, Yolanda Serrano Alonso, Carolina Sanchez Barranco

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Insects are a new source of fatty acids (FA), so they are considered a sustainable and environmentally friendly alternative for both animal feed and the human diet, and furthermore, their harvesting/rearing require a low-tech and low capital investment. For that reason, lipids obtained by insect breeding open interesting possibilities with alimentary and industrial purposes, i.e., the production of biodiesel. Particularly, certain insect species, especially during the larval stage, contain high proportions of fat which is highly dependent on their feed and stage of development. Among them, Hermetia illucens larvae can be bred on food wastes to produce fat- and protein-rich raw materials for food by-product management. So, insects can act as excellent bioconverters of organic waste to nutrient-rich materials. In this regard, the aim of the study was to evaluate the effects of fruit byproducts on the FA compositions of Tenebrio molitor, Zophoba morio, and H. illucens larvae. Firstly, oil was extracted with the green solvent ethyl acetate, and FA methyl ester was obtained and analyzed by GC to show the FA profile. In addition, the triacylglycerol (TAG) profile was obtained by HPLC. Dehydrated watermelon, tomato, and papaya by-products, as well as wheat-based control feed, were assayed. High FA content was reached by Z. morio larvae fed with all fruits; however, no differences were shown in lipid profile with any change. It is worth highlighting that both Z. morio and H. illucens could be selected as the best candidates for biodiesel production due to their high content of saturated FA. On the other hand, T. molitor larvae showed a higher content of monounsaturated FA than control larvae, whereas the n-6 polyunsaturated FA content decreased in larvae fed with fruits. This result indicates that the improvement of the FA profile of Tenebrio can depend on both the type of feeding and the intended use. The lipid profile of H. illucens larvae fed with papaya and tomato showed a slight increase in the content of α-linoleic acid (ALA, 18:3n3). This FA is the precursor of docosahexaenoic acid (DHA, 22:6n3), which plays an important role as a component of structural lipids in cell membranes as well as in the synthesis of eicosanoids, protecting and resolving. Also, it was evaluated the TAG profile of Z. morio larvae due to their highest oil content. The results showed a high oleic acid (OA, 18:1n9) content, which displays modulatory effects in a wide range of physiological functions, having anti-inflammatory and anti-atherogenic properties. In conclusion, this study clearly shows that Z. morio and H. illucens larvae constitute an alternative source of OA- and ALA-rich oils, respectively, which can be devoted for food use, as well as for using in the food and pharmaceutical industries, with agronomic implications. Finally, although the profile of Z. morio was not improved with fruit feeding, this kind of feeding could be used due to its low environmental impact.

Keywords: fatty acids, fruit byproducts, Hermetia illucens, Zophoba morio, Tenebrio molitor, insect rearing

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Authors: Leo Nnamdi Ozurumba-Dwight

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Messenger ribooxynucleic acid (mRNA) molecules are compositional, protein-based. These proteins, encoding mRNA molecules (which collectively connote the transcriptome), when analyzed by RNA sequencing (RNAseq), unveils the nature of gene expression in the RNA. The obtained gene expression provides clues of cellular traits and their dynamics in presentations. These can be studied in relation to function and responses. RNAseq is a practical concept in Genomics as it enables detection and quantitative analysis of mRNA molecules. Single cell and spatial transcriptomics both present varying avenues for expositions in genomic characteristics of single cells and pooled cells in disease conditions such as cancer, auto-immune diseases, hematopoietic based diseases, among others, from investigated biological tissue samples. Single cell transcriptomics helps conduct a direct assessment of each building unit of tissues (the cell) during diagnosis and molecular gene expressional studies. A typical technique to achieve this is through the use of a single-cell RNA sequencer (scRNAseq), which helps in conducting high throughput genomic expressional studies. However, this technique generates expressional gene data for several cells which lack presentations on the cells’ positional coordinates within the tissue. As science is developmental, the use of complimentary pre-established tissue reference maps using molecular and bioinformatics techniques has innovatively sprung-forth and is now used to resolve this set back to produce both levels of data in one shot of scRNAseq analysis. This is an emerging conceptual approach in methodology for integrative and progressively dependable transcriptomics analysis. This can support in-situ fashioned analysis for better understanding of tissue functional organization, unveil new biomarkers for early-stage detection of diseases, biomarkers for therapeutic targets in drug development, and exposit nature of cell-to-cell interactions. Also, these are vital genomic signatures and characterizations of clinical applications. Over the past decades, RNAseq has generated a wide array of information that is igniting bespoke breakthroughs and innovations in Biomedicine. On the other side, spatial transcriptomics is tissue level based and utilized to study biological specimens having heterogeneous features. It exposits the gross identity of investigated mammalian tissues, which can then be used to study cell differentiation, track cell line trajectory patterns and behavior, and regulatory homeostasis in disease states. Also, it requires referenced positional analysis to make up of genomic signatures that will be sassed from the single cells in the tissue sample. Given these two presented approaches to RNA transcriptomics study in varying quantities of cell lines, with avenues for appropriate resolutions, both approaches have made the study of gene expression from mRNA molecules interesting, progressive, developmental, and helping to tackle health challenges head-on.

Keywords: transcriptomics, RNA sequencing, single cell, spatial, gene expression.

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Authors: Zatovska T. V., Nesterova N. V., Baranova G. V., Zagorodnya S. D.

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In recent years, biosensor technologies based on the phenomenon of surface plasmon resonance (SPR) are becoming increasingly used in biology and medicine. Their application facilitates exploration in real time progress of binding of biomolecules and identification of agents that specifically interact with biologically active substances immobilized on the biosensor surface (biochips). Special attention is paid to the use of Biosensor analysis in determining the antibody-antigen interaction in the diagnostics of diseases caused by viruses and bacteria. According to WHO, the diseases that are caused by the herpes simplex virus (HSV), take second place (15.8%) after influenza as a cause of death from viral infections. Current diagnostics of HSV infection include PCR and ELISA assays. The latter allows determination the degree of immune response to viral infection and respective stages of its progress. In this regard, the searches for new and available diagnostic methods are very important. This work was aimed to develop Biosensor chip for detection of specific antibodies to HSV-1 in the human blood serum. The proteins of HSV1 (strain US) were used as antigens. The viral particles were accumulated in cell culture MDBK and purified by differential centrifugation in cesium chloride density gradient. Analysis of the HSV1 proteins was performed by polyacrylamide gel electrophoresis and ELISA. The protein concentration was measured using De Novix DS-11 spectrophotometer. The device for detection of antigen-antibody interactions was an optoelectronic two-channel spectrometer ‘Plasmon-6’, using the SPR phenomenon in the Krechman optical configuration. It was developed at the Lashkarev Institute of Semiconductor Physics of NASU. The used carrier was a glass plate covered with 45 nm gold film. Screening of human blood serums was performed using the test system ‘HSV-1 IgG ELISA’ (GenWay, USA). Development of Biosensor chip included optimization of conditions of viral antigen sorption and analysis steps. For immobilization of viral proteins 0.2% solution of Dextran 17, 200 (Sigma, USA) was used. Sorption of antigen took place at 4-8°C within 18-24 hours. After washing of chip, three times with citrate buffer (pH 5,0) 1% solution of BSA was applied to block the sites not occupied by viral antigen. It was found direct dependence between the amount of immobilized HSV1 antigen and SPR response. Using obtained biochips, panels of 25 positive and 10 negative for the content of antibodies to HSV-1 human sera were analyzed. The average value of SPR response was 185 a.s. for negative sera and from 312 to. 1264 a.s. for positive sera. It was shown that SPR data were agreed with ELISA results in 96% of samples proving the great potential of SPR in such researches. It was investigated the possibility of biochip regeneration and it was shown that application of 10 mM NaOH solution leads to rupture of intermolecular bonds. This allows reuse the chip several times. Thus, in this study biosensor chip for detection of specific antibodies to HSV1 was successfully developed expanding a range of diagnostic methods for this pathogen.

Keywords: biochip, herpes virus, SPR

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Authors: H. I. Obadiah, S. N. Wieser, E. A. Omudu, B. O. Atu, O. Byanet, L. Schnittger, M. Florin-Christensen

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Sarcocystis sp. are Apicomplexan protozoan parasites with a life cycle that involves a predator and a prey as final and intermediate hosts, respectively. In tissues of the intermediate hosts, the parasites produce sarcocysts that vary in size and morphology according to the species. When a suitable predator ingests sarcocyst-containing meat, the parasites are released in the intestine and undergo sexual reproduction producing infective sporocysts, which are excreted with the feces into the environment. The cycle is closed when a prey ingests sporocyst-contaminated water or pasture; the parasites gain access to the circulation, and eventually invade tissues and reproduce asexually yielding sarcocysts. Pig farming is a common practice in Nigeria as well as in many countries around the world. In addition to its importance as protein source, pork is also a source of several pathogens relevant to humans. In the case of Sarcocystis, three species have been described both in domestic and wild pigs, namely, S. miescheriana, S. porcifelis and S. suihominis. Humans can act both as final and aberrant intermediate hosts of S. suihominis, after ingesting undercooked sarcocyst-infested pork. Infections are usually asymptomatic but can be associated with inappetence, nausea, vomiting and diarrhea, or with muscle pain, fever, eosinophilia and bronchospasm, in humans acting as final or intermediate hosts, respectively. Moreover, excretion of infective forms with human feces leads to further dissemination of the infection. In this study, macroscopic sarcocysts of white color, oval shape and a size range of approximately 3-5 mm were observed in the skeletal muscle of a slaughtered pig in an abattoir in Makurdi, Benue State, Nigeria, destined to human consumption. Sarcocysts were excised and washed in distilled water, and genomic DNA was extracted using a commercial kit. The near-complete length of the 18S rRNA gene was analyzed after PCR amplification of two overlapping fragments, each of which were submitted to direct sequencing. In addition, the mitochondrial cytochrome oxidase (cox-1) gene was PCR-amplified and directly sequenced. Two phylogenetic trees containing the obtained sequences along with available relevant 18S rRNA and cox-1 sequences were constructed by neighbor joining after alignment, using the corresponding sequences of Toxoplasma gondii as outgroup. The results showed in both cases that the analyzed sequences grouped with S. suihominis with high bootstrap value, confirming the identity of this macroscopic sarcocyst-forming parasite as S. suihominis. To the best of our knowledge, these results represent the first demonstration of this parasite in pigs of Nigeria and the largest sarcocysts described so far for S. suihominis. The close proximity between pigs and humans in pig farms, and the frequent poor sanitary conditions in human dwellings strongly suggest that the parasite undergoes the sexual stages of its life cycle in humans as final hosts. These findings provide an important reference for the examination and control of Sarcocystis species in pigs of Nigeria.

Keywords: nigeria, pork, sarcocystis suihominis, zoonotic parasite

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Authors: Meena Agrawal, Chaitanya P. Agrawal

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The power systems are transforming and becoming smarter with innovations in technologies to enable embark simultaneously upon the sustainable energy needs, rising environmental concerns, economic benefits and quality requirements. The advantages provided by inter-connection of renewable energy resources are becoming more viable and dependable with the smart controlling technologies. The limitation of most renewable resources have their diversity and intermittency causing problems in power quality, grid stability, reliability, security etc. is being cured by these efforts. A necessitate of optimal energy management by intelligent Micro-Grids at the distribution end of the power system has been accredited to accommodate sustainable renewable Distributed Energy Resources on large scale across the power grid. All over the world Smart Grids are emerging now as foremost concern infrastructure upgrade programs. The hardware setup includes NI cRIO 9022, Compact Reconfigurable Input Output microcontroller board connected to the PC on a LAN router with three hardware modules. The Real-Time Embedded Controller is reconfigurable controller device consisting of an embedded real-time processor controller for communication and processing, a reconfigurable chassis housing the user-programmable FPGA, Eight hot-swappable I/O modules, and graphical LabVIEW system design software. It has been employed for signal analysis, controls and acquisition and logging of the renewable sources with the LabVIEW Real-Time applications. The employed cRIO chassis controls the timing for the module and handles communication with the PC over the USB, Ethernet, or 802.11 Wi-Fi buses. It combines modular I/O, real-time processing, and NI LabVIEW programmable. In the presented setup, the Analog Input Module NI 9205 five channels have been used for input analog voltage signals from renewable energy sources and NI 9227 four channels have been used for input analog current signals of the renewable sources. For switching actions based on the programming logic developed in software, a module having Electromechanical Relays (single-pole single throw) with 4-Channels, electrically isolated and LED indicating the state of that channel have been used for isolating the renewable Sources on fault occurrence, which is decided by the logic in the program. The module for Ethernet based Data Acquisition Interface ENET 9163 Ethernet Carrier, which is connected on the LAN Router for data acquisition from a remote source over Ethernet also has the module NI 9229 installed. The LabVIEW platform has been employed for efficient data acquisition, monitoring and control. Control logic utilized in program for operation of the hardware switching Related to Fault Relays has been portrayed as a flowchart. A communication system has been successfully developed amongst the sources and loads connected on different computers using Hypertext transfer protocol, HTTP or Ethernet Local Stacked area Network TCP/IP protocol. There are two main I/O interfacing clients controlling the operation of the switching control of the renewable energy sources over internet or intranet. The paper presents experimental results of the briefed setup for intelligent control of the micro-grid for renewable energy sources, besides the control of Micro-Grid with data acquisition and control hardware based on a microcontroller with visual program developed in LabVIEW.

Keywords: data acquisition and control, LabVIEW, microcontroller cRIO, Smart Micro-Grid

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Authors: S. F. Afzali, W. L. Wong

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The epizootic ulcerative syndrome (EUS) causes by the oomycete fungus, Aphanomyces invadans; known to be one of the infectious fish diseases for farmed and wild fishes in fresh and brackish-water from the Asia-pacific region, America and Africa. Although, EUS had been documented by the Office International des Epizooties (OIE) since 1995, hitherto, there is neither standard chemical agents that can be used for successful treatment of this destructive infection in the time of outbreak; nor available vaccine for prevention. Plant-based remedies in controlling fish diseases are gaining much attention recently as an alternative to chemical treatments, which possess negative effects to the environment and human. In present study, Sonneratia alba, a mangrove plant belongs to the Sonneratiaceae family, was screened in vitro and in vivo for its antifungal activity against A. invadans mycelium growth and its effects on fish innate immune system and disease resistant. The in vitro tests was performed using the disc diffusion methods with measurements of minimum inhibitory concentration (MIC) and inhibition zone. For in vivo study, the S. alba extract supplemented diets were administrated at 0.0, 1.0%, 3.0%, and 5.0% on healthy goldfish, Carassius auratus, which challenged with A. invadans zoospores (100 spores/ml). To compare the significant differences in the hematological and immunological parameters obtained from the experiments, the data were analysed using the SPSS. The methanol extract of S. alba effectively inhibited the mycelial growth of A. invadans at a minimum concentration of 1000 ppm for agar and filter paper diffusion experiments. In the agar diffusion test, 500 ppm of the extract inhibited the fungus mycelial growth up to 96 hours after exposure. The mycelial growth from the edge of the pre-inoculated A. invadans agar discs treated with S. alba extracts at concentrations of 100, 500 and 1000 ppm were 15, 8 and 0 mm respectively. The results of the filter paper disc test showed that the S. alba extract at its minimal inhibitory concentration (1000 ppm) has similar qualitative inhibitory effect as malachite green at 1 ppm and formalin at 250 ppm. According to the in vivo tests findings, in the infected fish fed with 3.0% and 5.0% supplementation diet, the numbers of white blood cell and myeloperoxidase activity significantly increased after the second week of treatment. Whilst the numbers of red blood cell significantly decreased in the infected fish fed with 0.0 and 1.0% supplementation diet. After the third week of feeding, significant increases in the total protein, albumin level, lysozyme activity were recorded in the infected fish fed with 3.0% and 5.0% supplementation diet. Also, the enriched diets increased the survival rate as compared to the untreated group that suffered from 90% mortality. The present study indicated that S. alba extract may inhibit the mycelial growth of A. invadans effectively, suggesting an alternative to other chemotherapeutic agents, which brought much environmental and health concerns to the public, for EUS treatment.

Keywords: fungal pathogen, goldfish, organic extract, treatment

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Authors: Yetti Marlida, Syukri Arif, Nadirman Haska

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Recently, alcohol fuels have been produced on industrial scales by fermentation of sugars derived from wheat, corn, sugar beets, sugar cane etc. The enzymatic hydrolysis of cellulosic materials to produce fermentable sugars has an enormous potential in meeting global bioenergy demand through the biorefinery concept, since agri-food processes generate millions of tones of waste each year (Xeros and Christakopoulos 2009) such as sugar cane baggase , wheat straw, rice straw, corn cob, and oil palm trunk. In fact oil palm trunk is one of the most abundant lignocellulosic wastes by-products worldwide especially come from Malaysia, Indonesia and Nigeria and provides an alternative substrate to produce useful chemicals such as bioethanol. Usually, from the ages 3 years to 25 years, is the economical life of oil palm and after that, it is cut for replantation. The size of trunk usually is 15-18 meters in length and 46-60 centimeters in diameter. The trunk after cutting is agricultural waste causing problem in elimination but due to the trunk contains about 42% cellulose, 34.4%hemicellulose, 17.1% lignin and 7.3% other compounds,these agricultural wastes could make value added products (Pumiput, 2006).This research was production of bioethanol from oil palm trunk via saccharafication by cocktail carbohydrases enzymes. Enzymatic saccharification of acid treated oil palm trunk was carried out in reaction mixture containing 40 g treated oil palm trunk in 200 ml 0.1 M citrate buffer pH 4.8 with 500 unit/kg amylase for treatment A: Treatment B: Treatment A + 500 unit/kg cellulose; C: treatment B + 500 unit/kgg xylanase: D: treatment D + 500 unit/kg ligninase and E: OPT without treated + 500 unit/kg amylase + 500 unit/kg cellulose + 500 unit/kg xylanase + 500 unit/kg ligninase. The reaction mixture was incubated on a water bath rotary shaker adjusted to 600C and 75 rpm. The samples were withdraw at intervals 12 and 24, 36, 48,60, and 72 hr. For bioethanol production in biofermentor of 5L the hydrolysis product were inoculated a loop of Saccharomyces cerevisiae and then incubated at 34 0C under static conditions. Samples are withdraw after 12, 24, 36, 48 and 72 hr for bioethanol and residual glucose. The results of the enzymatic hidrolysis (Figure1) showed that the treatment B (OPT hydrolyzed with amylase and cellulase) have optimum condition for glucose production, where was both of enzymes can be degraded OPT perfectly. The same results also reported by Primarini et al., (2012) reported the optimum conditions the hydrolysis of OPT was at concentration of 25% (w /v) with 0.3% (w/v) amylase, 0.6% (w /v) glucoamylase and 4% (w/v) cellulase. In the Figure 2 showed that optimum bioethanol produced at 48 hr after incubation,if time increased the biothanol decreased. According Roukas (1996), a decrease in the concentration of ethanol occur at excess glucose as substrate and product inhibition effects. Substrate concentration is too high reduces the amount of dissolved oxygen, although in very small amounts, oxygen is still needed in the fermentation by Saccaromyces cerevisiae to keep life in high cell concentrations (Nowak 2000, Tao et al. 2005). The results of the research can be conluded that the optimum enzymatic hydrolysis occured when the OPT added with amylase and cellulase and optimum bioethanol produced at 48 hr incubation using Saccharomyses cerevicea whereas 18.08 % bioethanol produced from glucose conversion. This work was funded by Directorate General of Higher Education (DGHE), Ministry of Education and Culture, contract no.245/SP2H/DIT.LimtabMas/II/2013

Keywords: oil palm trunk, enzymatic hydrolysis, saccharification

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Authors: Sandy Elsayed, Aya Mohamed, Noha Nassar

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Introduction: Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by social deficits and repetitive behavior. Multiple studies suggest that oxidative stress and neuroinflammation are key factors in the etiology of ASD and often associated with worsening of ASD-related behaviors. Nuclear factor erythroid 2-related factor 2 (NRF-2) is a transcription factor that promotes expression of antioxidant response element genes in oxidative stress. In ASD subjects, decreased expression of NRF-2 in frontal cortex shifted the redox homeostasis towards oxidative stress, and resulted in inflammation evidenced by elevation of nuclear factor kappa B (NF-κB) transcriptional activity. Dimethyl fumarate (DMF) is a NRF-2 activator that is used in the treatment of psoriasis and multiple sclerosis. It participates in the transcriptional control of inflammatory factors via inhibition of NF-κB and its downstream targets. This study aimed to investigate the role of DMF in alleviating the cognitive impairments and behavior deficits associated with ASD through mitigation of oxidative stress and inflammation in prenatal valproic acid (VPA) rat model of autism. Methods: Pregnant female Wistar rats received a single intraperitoneal injection of VPA (600 mg/kg) to induce autistic-like-behavioral and neurobiological alterations in their offspring. Chronic oral gavage of DMF (150mg/kg/day) started from postnatal day (PND) 24 till PND62 (39 days). Prenatal VPA exposure elicited autistic behaviors including decreased social interaction and stereotyped behavior. Social interaction was evaluated using three-chamber sociability test and calculation of sociability index (SI), while stereotyped repetitive behavior and anxiety associated with ASD were assessed using marble burying test (MBT). Biochemical analyses were done on prefrontal cortex homogenates including NRF-2, and NF-κB expression. Moreover, inducible nitric oxide synthase (iNOS) gene expression and tumor necrosis factor (TNF-) protein expression were evaluated as markers of inflammation. Results: Prenatal VPA elicited decreased social interaction shown by decreased SI compared to control group (p < 0.001) and DMF enhanced SI (p < 0.05). In MBT, prenatal injection of VPA manifested stereotyped behavior and enhanced number of buried marbles compared to control (p < 0.05) and DMF reduced the anxiety-related behavior in rats exhibiting ASD-like behaviors (p < 0.05). In prefrontal cortex, NRF-2 expression was downregulated in prenatal VPA model (p < 0.0001) and DMF reversed this effect (p < 0.0001). The inflammatory transcription factor NF-κB was elevated in prenatal VPA model (p < 0.0001) and reduced (p < 0.0001) upon NRF-2 activation by DMF. Prenatal VPA expressed higher levels of proinflammatory cytokine TNF- compared to control group (p < 0.0001) and DMF reduced it (p < 0.0001). Finally, the gene expression of iNOS was downregulated upon NRF-2 activation by DMF (p < 0.01). Conclusion: This study proposes that DMF is a potential agent that can be used to ameliorate autistic-like-changes through NRF-2 activation along with NF-κB downregulation and therefore, it is a promising novel therapy for ASD.

Keywords: autism spectrum disorders, dimethyl fumarate, neuroinflammation, NRF-2

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Authors: B. Atika, S. Lehmann, E. Wimmer

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The defense strategy is very common in the insect world. Defensive substances play a wide variety of functions for beetles, such as repellents, toxicants, insecticides, and antimicrobics. Beetles react to predators, invaders, and parasitic microbes with the release of toxic and repellent substances. Defensive substances are directed against a large array of potential target organisms or may function for boiling bombardment or as surfactants. Usually, Coleoptera biosynthesize and store their defensive compounds in a complex secretory organ, known as odoriferous defensive stink glands. The red flour beetle, Tribolium castaneum (Coleoptera: Tenebrionidae), uses these glands to produce antimicrobial p-benzoquinones and 1-alkenes. In the past, the morphology of stink gland has been studied in detail in tenebrionid beetles; however, very little is known about the genes that are involved in the production of gland secretion. In this study, we studied a subset of genes that are essential for the benzoquinone production in red flour beetle. In the first phase, we selected 74 potential candidate genes from a genome-wide RNA interference (RNAi) knockdown screen named 'iBeetle.' All these 74 candidate genes were functionally characterized by RNAi-mediated gene knockdown. Therefore, they were selected for a subsequent gas chromatography-mass spectrometry (GC-MS) analysis of secretion volatiles in respective RNAi knockdown glands. 33 of them were observed to alter the phenotype of stink gland. In the GC-MS analysis, 7 candidate genes were noted to display a strongly altered gland, in terms of secretion color and chemical composition, upon knockdown, showing their key role in the biosynthesis of gland secretion. Morphologically altered stink glands were found for odorant receptor and protein kinase superfamily. Subsequent GC-MS analysis of secretion volatiles revealed reduced benzoquinone levels in LIM domain, PDZ domain, PBP/GOBP family knockdowns and a complete lack of benzoquinones in the knockdown of sulfatase-modifying factor enzyme 1, sulfate transporter family. Based on stink gland transcriptome data, we analyzed the function of sulfatase-modifying factor enzyme 1 and sulfate transporter family via RNAi-mediated gene knockdowns, GC-MS, in situ hybridization, and enzymatic activity assays. Morphologically altered stink glands were noted in knockdown of both these genes. Furthermore, GC-MS analysis of secretion volatiles showed a complete lack of benzoquinones in the knockdown of these two genes. In situ hybridization showed that these two genes are expressed around the vesicle of certain subgroup of secretory stink gland cells. Enzymatic activity assays on stink gland tissue showed that these genes are involved in p-benzoquinone biosynthesis. These results suggest that sulfatase-modifying factor enzyme 1 and sulfate transporter family play a role specifically in benzoquinone biosynthesis in red flour beetles.

Keywords: Red Flour Beetle, defensive stink gland, benzoquinones, sulfate transporter, sulfatase-modifying factor enzyme 1

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Authors: Nicholas Mavrogiannis, Francesca Crivellari, Zachary Gagnon

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Development of low-cost, rapid, sensitive and portable biosensing systems are important for the detection and prevention of disease in developing countries, biowarfare/antiterrorism applications, environmental monitoring, point-of-care diagnostic testing and for basic biological research. Currently, the most established commercially available and widespread assays for portable point of care detection and disease testing are paper-based dipstick and lateral flow test strips. These paper-based devices are often small, cheap and simple to operate. The last three decades in particular have seen an emergence in these assays in diagnostic settings for detection of pregnancy, HIV/AIDS, blood glucose, Influenza, urinary protein, cardiovascular disease, respiratory infections and blood chemistries. Such assays are widely available largely because they are inexpensive, lightweight, and portable, are simple to operate, and a few platforms are capable of multiplexed detection for a small number of sample targets. However, there is a critical need for sensitive, quantitative and multiplexed detection capabilities for point-of-care diagnostics and for the detection and prevention of disease in the developing world that cannot be satisfied by current state-of-the-art paper-based assays. For example, applications including the detection of cardiac and cancer biomarkers and biothreat applications require sensitive multiplexed detection of analytes in the nM and pM range, and cannot currently be satisfied with current inexpensive portable platforms due to their lack of sensitivity, quantitative capabilities and often unreliable performance. In this talk, inexpensive label-free biomolecular detection at liquid interfaces using a newly discovered electrokinetic phenomenon known as fluidic dielectrophoresis (fDEP) is demonstrated. The electrokinetic approach involves exploiting the electrical mismatches between two aqueous liquid streams forced to flow side-by-side in a microfluidic T-channel. In this system, one fluid stream is engineered to have a higher conductivity relative to its neighbor which has a higher permittivity. When a “low” frequency (< 1 MHz) alternating current (AC) electrical field is applied normal to this fluidic electrical interface the fluid stream with high conductivity displaces into the low conductive stream. Conversely, when a “high” frequency (20MHz) AC electric field is applied, the high permittivity stream deflects across the microfluidic channel. There is, however, a critical frequency sensitive to the electrical differences between each fluid phase – the fDEP crossover frequency – between these two events where no fluid deflection is observed, and the interface remains fixed when exposed to an external field. To perform biomolecular detection, two streams flow side-by-side in a microfluidic T-channel: one fluid stream with an analyte of choice and an adjacent stream with a specific receptor to the chosen target. The two fluid streams merge and the fDEP crossover frequency is measured at different axial positions down the resulting liquid

Keywords: biodetection, fluidic dielectrophoresis, interfacial polarization, liquid interface

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Authors: Enya Trenholm-Jensen

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This paper employs various semiotic methodologies to examine the mechanism of soft power. Soft power refers to a country’s global reputation and their ability to leverage that reputation to achieve certain aims. South Korea has invested heavily in their soft power strategy for a multitude of predominantly historical and geopolitical reasons. On account of this investment and the global prominence of their strategy, South Korea was considered to be the optimal candidate for the aims of this investigation. Having isolated the entertainment industry as one of the most heavily funded segments of the South Korean soft power strategy, the analysis restricted itself to this sector. Within this industry, two entertainment products were selected as case studies. The case studies were chosen based on commercial success according to metrics such as streams, purchases, and subsequent revenue. This criterion was deemed to be the most objective and verifiable indicator of the products general appeal. The entertainment products which met the chosen criterion were Netflix’ “Squid Game” and BTS’ hit single “Butter”. The methodologies employed were chosen according to the medium of the entertainment products. For “Squid Game,” an aesthetic analysis was carried out to investigate how multi- layered meanings were mobilized in a show popularized by its visual grammar. To examine “Butter”, both music semiology and linguistic analysis were employed. The music section featured an analysis underpinned by denotative and connotative music semiotic theories borrowing from scholars Theo van Leeuwen and Martin Irvine. The linguistic analysis focused on stance and semantic fields according to scholarship by George Yule and John W. DuBois. The aesthetic analysis of the first case study revealed intertextual references to famous artworks, which served to augment the emotional provocation of the Squid Game narrative. For the second case study, the findings exposed a set of musical meaning units arranged in a patchwork of familiar and futuristic elements to achieve a song that existed on the boundary between old and new. The linguistic analysis of the song’s lyrics found a deceptively innocuous surface level meaning that bore implications for authority, intimacy, and commercial success. Whether through means of visual metaphor, embedded auditory associations, or linguistic subtext, the collective findings of the three analyses exhibited a desire to conjure a form of positive arousal in the spectator. In the synthesis section, this process is likened to that of branding. Through an exploration of branding, the entertainment products can be understood as cogs in a larger operation aiming to create positive associations to Korea as a country and a concept. Limitations in the form of a timeframe biased perspective are addressed, and directions for future research are suggested. This paper employs semiotic methodologies to examine two entertainment products as mechanisms of soft power. Through means of visual metaphor, embedded auditory associations, or linguistic subtext, the findings reveal a desire to conjure positive arousal in the spectator. The synthesis finds similarities to branding, thus positioning the entertainment products as cogs in a larger operation aiming to create positive associations to Korea as a country and a concept.

Keywords: BTS, cognitive semiotics, entertainment, soft power, south korea, squid game

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Authors: Laura Norfolk, Georgina Zimbitas, Jan Sefcik, Sarah Staniland

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Magnetite nanoparticles (MNP) have been an area of increasing research interest due to their extensive applications in industry, such as in carbon capture, water purification, and crucially, the biomedical industry. The use of MNP in the biomedical industry is rising, with studies on their effect as Magnetic resonance imaging contrast agents, drug delivery systems, and as hyperthermic cancer treatments becoming prevalent in the nanomaterial research community. Particles used for biomedical purposes must meet stringent criteria; the particles must have consistent shape and size between particles. Variation between particle morphology can drastically alter the effective surface area of the material, making it difficult to correctly dose particles that are not homogeneous. Particles of defined shape such as octahedral and cubic have been shown to outperform irregular shaped particles in some applications, leading to the need to synthesize particles of defined shape. In nature, highly homogeneous MNP are found within magnetotactic bacteria, a unique bacteria capable of producing magnetite nanoparticles internally under ambient conditions. Biomineralisation proteins control the properties of the MNPs, enhancing their homogeneity. One of these proteins, Mms6, has been successfully isolated and used in vitro as an additive in room-temperature co-precipitation reactions (RTCP) to produce particles of defined mono-dispersed size & morphology. When considering future industrial scale-up it is crucial to consider the costs and feasibility of an additive, as an additive that is not readily available or easily synthesized at a competitive price will not be sustainable. As such, additives selected for this research are inspired by the functional groups of biomineralisation proteins, but cost-effective, environmentally friendly, and compatible with scale-up. Diethylenetriamine (DETA), triethylenetetramine (TETA), tetraethylenepentamine (TEPA), and pentaethylenehexamine (PEHA) have been successfully used in RTCP to modulate the properties of particles synthesized, leading to the formation of octahedral nanoparticles with no use of organic solvents, heating, or toxic precursors. By extending this principle to a fluidic system, ongoing research will reveal whether the amine additives can also exert morphological control in an environment which is suited toward higher particle yield. Two fluidic systems have been employed; a peristaltic turbulent flow mixing system suitable for the rapid production of MNP, and a macrofluidic system for the synthesis of tailored nanomaterials under a laminar flow regime. The presence of the amine additives in the turbulent flow system in initial results appears to offer similar morphological control as observed under RTCP conditions, with higher proportions of octahedral particles formed. This is a proof of concept which may pave the way to green synthesis of tailored MNP on an industrial scale. Mms6 and amine additives have been used in the macrofluidic system, with Mms6 allowing magnetite to be synthesized at unfavourable ferric ratios, but no longer influencing particle size. This suggests this synthetic technique while still benefiting from the addition of additives, may not allow additives to fully influence the particles formed due to the faster timescale of reaction. The amine additives have been tested at various concentrations, the results of which will be discussed in this paper.

Keywords: bioinspired, green synthesis, fluidic, magnetite, morphological control, scale-up

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Authors: Shatarupa Thakurta Roy

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Along the limelight of mainstream academic practices in Indian art, exist a significant lot of habitual art practices that are mutually susceptible in their contemporary forms. Narrative folk paintings of regional India has successfully dispersed to its audience social messages through pulsating pictures and orations. The paper consists of images from narrative scroll paintings on ‘Ramayana’ theme from various neighboring states as well as districts in India, describing their subtle differences in style of execution, method, and use of material. Despite sharing commonness in the choice of subject matter, habitual and ceremonial Indian folk art in its formative phase thrived within isolated locations to yield in remarkable variety in the art styles. The differences in style took place district wise, cast wise and even gender wise. An open flow is only evident in the contemporary expressions as a result of substantial changes in social structures, mode of communicative devices, cross-cultural exposures and multimedia interactivities. To decipher the complex nature of popular cultural taste of contemporary India it is important to categorically identify its root in vernacular symbolism. The realization of modernity through European primitivism was rather elevated as a perplexed identity in Indian cultural margin in the light of nationalist and postcolonial ideology. To trace the guiding factor that has still managed to obtain ‘Indianness’ in today’s Indian art, researchers need evidences from the past that are yet to be listed in most instances. They are commonly created on ephemeral foundations. The artworks are also found in endangered state and hence, not counted much friendly for frequent handling. The museums are in dearth of proper technological guidelines to preserve them. Even though restoration activities are emerging in the country, the existing withered and damaged artworks are in threat to perish. An immediacy of digital achieving is therefore envisioned as an alternative to save this cultural legacy. The method of this study is, two folded. It primarily justifies the richness of the evidences by conducting categorical aesthetic analysis. The study is supported by comments on the stylistic variants, thematic aspects, and iconographic identities alongside its anthropological and anthropomorphic significance. Further, it explores the possible ways of cultural preservation to ensure cultural sustainability that includes technological intervention in the form of digital transformation as an altered paradigm for better accessibility to the available recourses. The study duly emphasizes on visual description in order to culturally interpret and judge the rare visual evidences following Feldman’s four-stepped method of formal analysis combined with thematic explanation. A habitual design that emerges and thrives within complex social circumstances may experience change placing its principle philosophy at risk by shuffling and altering with time. A tradition that respires in the modern setup struggles to maintain timeless values that operate its creative flow. Thus, the paper hypothesizes the survival and further growth of this practice within the dynamics of time and concludes in realization of the urgency to transform the implicitness of its knowledge into explicit records.

Keywords: aesthetic, identity, implicitness, paradigm

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Authors: Laura E. Hall, Joel Monárrez-Espino, Luz María Tejada Tayabas

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College students are at risk of weight gain and poor dietary habits, and health behaviours established during this period have been shown to track into midlife. They may therefore be an important target group for health promotion strategies, yet there is a lack of literature regarding dietary intakes and associated factors in this group, particularly in middle-income countries such as Mexico. The aim of this exploratory research was to describe and compare reported dietary intakes among nursing and nutrition college students at two public universities in Mexico, and to explore the relationship between demographic, behavioural and other health-related factors and the risk of low diet quality. Mexican college students (n=444) majoring in nutrition or nursing at two urban universities completed questionnaires regarding dietary and health-related behaviours and risks. Dietary intake was assessed via 24-hour recall. Weight, height and abdominal circumference were measured. Descriptive statistics were reported and nutrient intakes were compared between colleges and study tracks using Student’s t tests, odds ratios and Pearson chi square tests. Two dietary quality scores were constructed to explore the relationship between demographic, behavioural and other health-related factors and the diet quality scores using binary logistic regression. Analysis was performed using SPSS statistics, with differences considered statistically significant at p<0.05. The response rate to the survey was 91%. When macronutrients were considered as a percentage of total energy, the majority of students had protein intakes within recommended ranges, however one quarter of students had carbohydrate and fat intakes exceeding recommended levels. Three quarters had fibre intakes that were below recommendations. More than half of the students reported intakes of magnesium, zinc, vitamin A, folate and vitamin E that were below estimated average requirements. Students studying nutrition reported macronutrient and micronutrient intakes that were more compliant with recommendations compared to nursing students, and students studying in central-north Mexico were more compliant than those studying in southeast Mexico. Breakfast skipping (Adjusted Odds Ratio (OR) = 5.3; 95% Confidence Interval (CI) = 1.2-22.7), risk of anxiety (OR = 2.3; CI = 1.3-4.4), and university location (OR = 1.6; CI = 1.03-2.6) were associated with a greater risk of having a low macronutrient score. Caloric intakes <1800kcal (OR = 5.8; CI = 3.5-9.7), breakfast skipping (OR = 3.7; CI = 1.4-10.3), vigorous exercise ≤1h/week (OR = 2.6; CI = 1.3-5.2), soda consumption >250mls/day (OR = 2.0; CI = 1.2-3.3), unhealthy diet perception (OR = 1.9; CI = 1.2-3.0), and university location (OR = 1.8; CI = 1.1-2.8) were significantly associated with greater odds of having a low micronutrient score. College students studying nursing and nutrition did not report ideal diets, and these students should not be overlooked in public health interventions. Differences in dietary intakes between universities and study tracks were evident, with more favourable profiles evident in nutrition compared to nursing, and North-central compared to Southeast students. Further, demographic, behavioural and other health-related factors were associated with diet quality scores, warranting further research.

Keywords: college student, diet quality, nutrient intake, young adult

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Authors: Marthinus Conradie

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The conversational dynamics of democratically orientated deliberation continue to stimulate critical scholarship for its potential to bolster robust engagement between different sections of pluralist societies. Several axes of deliberation that have attracted academic attention include face-to-face vs. online interaction, and citizen-to-citizen communication vs. engagement between citizens and political elites. In all these areas, numerous researchers have explored deliberative procedures aimed at achieving instrumental goals such a securing consensus on policy issues, against procedures that prioritise expressive outcomes such as broadening the range of argumentative repertoires that discursively construct and mediate specific political issues. The study that informs this paper, works in the latter stream. Drawing its data from the reader-comments section of a South African broadsheet newspaper, the study investigates online, citizen-to-citizen deliberation by analysing the discursive practices through which competing understandings of social problems are articulated and contested. To advance this agenda, the paper deals specifically with user-generated comments posted in response to news stories on questions of race and racism in South Africa. The analysis works to discern and interpret the various sets of discourse practices that shape how citizens deliberate contentious political issues, especially racism. Since the website in question is designed to encourage the critical comparison of divergent interpretations of news events, without feeding directly into national policymaking, the study adopts an analytic framework that traces how citizens articulate arguments, rather than the instrumental effects that citizen deliberations might exert on policy. The paper starts from the argument that such expressive interactions are particularly crucial to current trends in South African politics, given that the precise nature of race and racism remain contested and uncertain. Centred on a sample of 2358 conversational moves in 814 posts to 18 news stories emanating from issues of race and racism, the analysis proceeds in a two-step fashion. The first stage conducts a qualitative content analysis that offers insights into the levels of reciprocity among commenters (do readers engage with each other or simply post isolated opinions?), as well as the structures of argumentation (do readers support opinions by citing evidence?). The second stage involves a more fine-grained discourse analysis, based on a theorisation of argumentation that delineates it into three components: opinions/conclusions, evidence/data to support opinions/conclusions and warrants that explicate precisely how evidence/data buttress opinions/conclusions. By tracing the manifestation and frequency of specific argumentative practices, this study contributes to the archive of research currently aggregating around the practices that characterise South Africans’ engagement with provocative political questions, especially racism and racial inequity. Additionally, the study also contributes to recent scholarship on the affordances of Web 2.0 software by eschewing a simplistic bifurcation between cyber-optimist vs. pessimism, in favour of a more nuanced and context-specific analysis of the patterns that structure online deliberation.

Keywords: online deliberation, discourse analysis, qualitative content analysis, racism

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Authors: E. Tan, O. McKay, T. Clarnette T., D. Croagh

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Background: Historically with pancreatic trauma, complete disruption of the main pancreatic duct (MPD), classified as Grade IV-V by the American Association for the Surgery of Trauma (AAST), necessitated a damage-control laparotomy. This was to avoid mortality, shorten diet upgrade timeframe, and hence shorter length of stay. However, acute pancreatic resection entailed complications of pancreatic fistulas and leaks. With the advance of imaging-guided interventions, non-operative management such as percutaneous and transpapillary drainage of traumatic peripancreatic collections have been trialled favourably. The aim of this case series is to evaluate the efficacy of endoscopic ultrasound-guided (EUS) transmural drainage in managing traumatic peripancreatic collections as a less invasive alternative to traditional approaches. This study also highlights the importance of anatomical knowledge regarding peripancreatic collection’s common location in the lesser sac, the pancreas relationship to adjacent organs, and the formation of the main pancreatic duct in regards to the feasibility of therapeutic internal drainage. Methodology: A retrospective case series was conducted at a single tertiary endoscopy unit, analysing patient data over a 5-year period. Inclusion criteria outlined patients age 5 to 80-years-old, traumatic pancreatic injury of at least Grade IV and haemodynamic stability. Exclusion criteria involved previous episodes of pancreatitis or abdominal trauma. Patient demographics and clinicopathological characteristics were retrospectively collected. Results: The study identified 7 patients with traumatic pancreatic injuries that were managed from 2018-2022; age ranging from 5 to 34 years old, with majority being female (n=5). Majority of the mechanisms of trauma were a handlebar injury (n=4). Diagnosis was confirmed with an elevated lipase and computerized tomotography (CT) confirmation of proximal pancreatic transection with MPD disruption. All patients sustained an isolated single organ grade IV pancreatic injury, except case 4 and 5 with other intra-abdominal visceral Grade 1 injuries. 6 patients underwent early ERCP-guided transpapillary drainage with 1 being unsuccessful for pancreatic duct stent insertion (case 1) and 1 complication of stent migration (case 2). Surveillance imaging post ERCP showed the stents were unable to bridge the disrupted duct and development of symptomatic collections with an average size of 9.9cm. Hence, all patients proceeded to EUS-guided transmural drainage, with 2/7 patients requiring repeat drainages (case 6 and 7). Majority (n=6) had a cystogastrostomy, whilst 1 (case 6) had a cystoenterostomy due to feasibility of the peripancreatic collection being adjacent to duodenum rather than stomach. However, case 6 subsequently required repeat EUS-guided drainage with cystogastrostomy for ongoing collections. Hence all patients avoided initial laparotomy with an average index length of stay of 11.7 days. Successful transmural drainage was demonstrated, with no long-term complications of pancreatic insufficiency; except for 1 patient requiring a distal pancreatectomy at 2 year follow-up due to chronic pain. Conclusion: The early results of this series support EUS-guided transmural drainage as a viable management option for traumatic peripancreatic collections, showcasing successful outcomes, minimal complications, and long-term efficacy in avoiding surgical interventions. More studies are required before the adoption of this procedure as a less invasive and complication-prone management approach for traumatic peripancreatic collections.

Keywords: endoscopic ultrasound, cystogastrostomy, pancreatic trauma, traumatic peripancreatic collection, transmural drainage

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Authors: Mohsen Abdelrazek Khorshid Ali Selim

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Disadvantages and errors of current concrete reinforcement methodsL: Current concrete reinforcement methods are adopted in most parts of the world in their various doctrines and names. They adopt the so-called concrete slab system, where these slabs are semi-independent and isolated from each other and from the surrounding environment of concrete columns or beams, so that the reinforcing steel does not cross from one slab to another or from one slab to adjacent columns. It or the beams surrounding it and vice versa are only a few centimeters and no more. The same applies exactly to the concrete columns that support the building, where the reinforcing steel does not extend from the slabs or beams to the inside of the columns or vice versa except for a few centimeters and no more, just as the reinforcing steel does not extend from inside the column at the top. The ceiling is only a few centimetres, and the same thing is literally repeated in the concrete beams that connect the columns and separate the slabs, where the reinforcing steel does not cross from one beam to another or from one beam to the slabs or columns adjacent to it and vice versa, except for a few centimeters, which makes the basic building elements of columns, slabs and beams They all work in isolation from each other and from the environment surrounding them from all sides. This traditional method of reinforcement may be valid and lasting in geographical areas that are not exposed to earthquakes and earthquakes, where all the loads and tensile forces in the building are constantly directed vertically downward due to gravity and are borne directly by the vertical reinforcement of the building. However, in the case of earthquakes and earthquakes, the loads and tensile forces in the building shift from the vertical direction to the horizontal direction at an angle of inclination that depends on the strength of the earthquake, and most of them are borne by the horizontal reinforcement extending between the basic elements of the building, such as columns, slabs and beams, and since the crossing of the reinforcement between each of the columns, slabs and beams between them And each other, and vice versa, does not exceed several centimeters. In any case, the tensile strength, cohesion and bonding between the various parts of the building are very weak, which causes the buildings to disintegrate and collapse in the horrific manner that we saw in the earthquake in Turkey and Syria in February 2023, which caused the collapse of tens of thousands of buildings in A few seconds later, it left more than 50,000 dead, hundreds of thousands injured, and millions displaced. Description of the new earthquake-resistant model: The idea of the new model in the reinforcement of concrete buildings and constructions is based on the theory that we have formulated as follows: [The tensile strength, cohesion and bonding between the basic parts of the concrete building (columns, beams and slabs) increases as the lengths of the reinforcing steel bars increase and they extend and branch and the different parts of the building share them with each other.] . In other words, the strength, solidity, and cohesion of concrete buildings increase and they become resistant to earthquakes as the lengths of the reinforcing steel bars increase, extend, branch, and share with the various parts of the building, such as columns, beams, and slabs. That is, the reinforcing skewers of the columns must extend in their lengths without cutting to cross from one floor to another until their end. Likewise, the reinforcing skewers of the beams must extend in their lengths without cutting to cross from one beam to another. The ends of these skewers must rest at the bottom of the columns adjacent to the beams. The same thing applies to the reinforcing skewers of the slabs where they must These skewers should be extended in their lengths without cutting to cross from one tile to another, and the ends of these skewers should rest either under the adjacent columns or inside the beams adjacent to the slabs as follows: First, reinforce the columns: The columns have the lion's share of the reinforcing steel in this model in terms of type and quantity, as the columns contain two types of reinforcing bars. The first type is large-diameter bars that emerge from the base of the building, which are the nerves of the column. These bars must extend over their normal length of 12 meters or more and extend to a height of three floors, if desired. In raising other floors, bars with the same diameter and the same length are added to the top after the second floor. The second type is bars with a smaller diameter, and they are the same ones that are used to reinforce beams and slabs, so that the bars that reinforce the beams and slabs facing each column are bent down inside this column and along the entire length of the column. This requires an order. Most engineers do not prefer it, which is to pour the entire columns and pour the roof at once, but we prefer this method because it enables us to extend the reinforcing bars of both the beams and slabs to the bottom of the columns so that the entire building becomes one concrete block that is cohesive and resistant to earthquakes. Secondly, arming the cameras: The beams' reinforcing skewers must also extend to a full length of 12 meters or more without cutting. The ends of the skewers are bent and dropped inside the column at the beginning of the beam to its bottom. Then the skewers are extended inside the beam so that their other end falls under the facing column at the end of the beam. The skewers may cross over the head of a column. Another passes through another adjacent beam and rests at the bottom of a third column, according to the lengths of each of the skewers and beams. Third, reinforcement of slabs: The slab reinforcing skewers must also extend their entire length, 12 meters or more, without cutting, distinguishing between two cases. The first case is the skewers opposite the columns, and their ends are dropped inside one of the columns. Then the skewers cross inside the adjacent slab and their other end falls below the opposite column. The skewers may cross over The head of the adjacent column passes through another adjacent slab and rests at the bottom of a third column, according to the dimensions of the slabs and the lengths of the skewers. The second case is the skewers opposite the beams, and their ends must be bent in the form of a square or rectangle according to the dimensions of the beam’s width and height, and this square or rectangle is dropped inside the beam at the beginning of the slab, and it serves as The skewers are for the beams, then the skewers are extended along the length of the slab, and at the end of the slab, the skewers are bent down to the bottom of the adjacent beam in the shape of the letter U, after which the skewers are extended inside the adjacent slab, and this is repeated in the same way inside the other adjacent beams until the end of the skewer, then it is bent downward in the form of a square or rectangle inside the beam, as happened. In its beginning.

Keywords: earthquake resistant buildings, earthquake resistant concrete constructions, new technology for reinforcement of concrete buildings, new technology in concrete reinforcement

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Authors: Andrés Durán, Brian Castellanos, Marta Quicazán, Carlos Zuluaga-Domínguez

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Bee-pollen is an apicultural-derived food product, with a growing appreciation among consumers given the remarkable nutritional and functional composition, in particular, protein (24%), dietary fiber (15%), phenols (15 – 20 GAE/g) and carotenoids (600 – 900 µg/g). These properties are given by the geographical and climatic characteristics of the region where it is collected. There are several countries recognized by their pollen production, e.g. China, United States, Japan, Spain, among others. Beekeepers use traps in the entrance of the hive where bee-pollen is collected. After the removal of foreign particles and drying, this product is ready to be marketed. However, in countries located along the equator, the absence of seasons and a constant tropical climate throughout the year favors a more rapid spoilage condition for foods with elevated water activity. The climatic conditions also trigger the proliferation of microorganisms and insects. This, added to the factor that beekeepers usually do not have adequate processing systems for bee-pollen, leads to deficiencies in the quality and safety of the product. In contrast, the Andean region of South America, lying on equator, typically has a high production of bee-pollen of up to 36 kg/year/hive, being four times higher than in countries with marked seasons. This region is also located in altitudes superior to 2500 meters above sea level, having extremes sun ultraviolet radiation all year long. As a mechanism of defense of radiation, plants produce more secondary metabolites acting as antioxidant agents, hence, plant products such as bee-pollen contain remarkable more phenolics and carotenoids than collected in other places. Considering this, the improvement of bee-pollen processing facilities by technical modifications and the implementation of an integrated cleaning and drying system for the product in an apiary in the area was proposed. The beehives were modified through the installation of alternative bee-pollen traps to avoid sources of contamination. The processing facility was modified according to considerations of Good Manufacturing Practices, implementing the combined use of a cabin dryer with temperature control and forced airflow and a greenhouse-type solar drying system. Additionally, for the separation of impurities, a cyclone type system was implemented, complementary to a screening equipment. With these modifications, a decrease in the content of impurities and the microbiological load of bee-pollen was seen from the first stages, principally with a reduction of the presence of molds and yeasts and in the number of foreign animal origin impurities. The use of the greenhouse solar dryer integrated to the cabin dryer allowed the processing of larger quantities of product with shorter waiting times in storage, reaching a moisture content of about 6% and a water activity lower than 0.6, being appropriate for the conservation of bee-pollen. Additionally, the contents of functional or nutritional compounds were not affected, even observing an increase of up to 25% in phenols content and a non-significant decrease in carotenoids content and antioxidant activity.

Keywords: beekeeping, drying, food processing, food safety

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Authors: Ricardo Godoy, Herbert Venthur, Hector Jimenez, Andres Quiroz, Ana Mutis

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In agriculture, grape production has great economic importance at global level, considering that in 2013 it reached 7.4 million hectares (ha) covered by plantations of this fruit worldwide. Chile is the number one exporter in the world with 800,000 tons. However, these values have been threatened by the attack of the grapevine moth, Lobesia botrana (Denis & Schiffermuller) (Lepidoptera: Tortricidae), since its detection in 2008. Nowadays, the use of semiochemicals, in particular the major component of the sex pheromone, (E,Z)-7.9-dodecadienil acetate, are part of mating disruption methods to control L. botrana. How insect pests can recognize these molecules, is being part of huge efforts to deorphanize their olfactory mechanism at molecular level. Thus, an interesting group of proteins has been identified in the antennae of insects, where odorant-binding proteins (OBPs) are known by transporting molecules to odorant receptors (ORs) and a co-receptor (ORCO) causing a behavioral change in the insect. Other proteins such as chemosensory proteins (CSPs), ionotropic receptors (IRs), odorant degrading enzymes (ODEs) and sensory neuron membrane proteins (SNMPs) seem to be involved, but few studies have been performed so far. The above has led to an increasing interest in insect communication at a molecular level, which has contributed to both a better understanding of the olfaction process and the design of new pest management strategies. To date, it has been reported that the ORs can detect one or a small group of odorants in a specific way. Therefore, the objective of this study is the identification of genes that encode these ORs using the antennal transcriptome of L. botrana. Total RNA was extracted for females and males of L. botrana, and the antennal transcriptome sequenced by Next Generation Sequencing service using an Illumina HiSeq2500 platform with 50 million reads per sample. Unigenes were assembled using Trinity v2.4.0 package and transcript abundance was obtained using edgeR. Genes were identified using BLASTN and BLASTX locally installed in a Unix system and based on our own Tortricidae database. Those Unigenes related to ORs were characterized using ORFfinder and protein Blastp server. Finally, a phylogenetic analysis was performed with the candidate amino acid sequences for LbotORs including amino acid sequences of other moths ORs, such as Bombyx mori, Cydia pomonella, among others. Our findings suggest 61 genes encoding ORs and one gene encoding an ORCO in both sexes, where the greatest difference was found in the OR6 because of the transcript abundance according to the value of FPKM in females and males was 1.48 versus 324.00. In addition, according to phylogenetic analysis OR6 is closely related to OR1 in Cydia pomonella and OR6, OR7 in Epiphyas postvittana, which have been described as pheromonal receptors (PRs). These results represent the first evidence of ORs present in the antennae of L. botrana and a suitable starting point for further functional studies with selected ORs, such as OR6, which is potentially related to pheromonal recognition.

Keywords: antennal transcriptome, lobesia botrana, odorant receptors (ORs), phylogenetic analysis

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Authors: Sithandazile Masuku

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Introduction: Public health approaches to offender mental health are driven by international policies and frameworks in response to the disproportionately large representation of people with mental health problems within the offender pathway compared to the general population. Public health service innovations include mental health courts in the US, restorative models in Singapore and, liaison and diversion services in Australia, the UK, and some other European countries. Mental health nurses are at the forefront of offender health service innovations. In the U.K. context, police custody has been identified as an early point within the offender pathway where nurses can improve outcomes by offering assessments and share information with criminal justice partners. This scope of nursing practice has introduced challenges related to skills and support required for nurses working at the interface of health and the criminal justice system. Parallel literature exploring experiences of nurses working in forensic settings suggests the presence of compassion fatigue, burnout and vicarious trauma that may impede risk harm to the nurses in these settings. Published research explores mainly service-level outcomes including monitoring of figures indicative of a reduction in offending behavior. There is minimal research exploring the experiences of liaison and diversion nurses who are situated away from a supportive clinical environment and engaged in complex autonomous decision-making. Aim: This paper will share qualitative findings (in progress) from a PhD study that aims to explore the experiences of liaison and diversion nurses in one service in the U.K. Methodology: This is a qualitative interview study conducted using an Interpretative Phenomenological Analysis to gain an in-depth analysis of lived experiences. Methods: A purposive sampling technique was used to recruit n=8 mental health nurses registered with the UK professional body, Nursing and Midwifery Council, from one UK Liaison and Diversion service. All participants were interviewed online via video call using semi-structured interview topic guide. Data were recorded and transcribed verbatim. Data were analysed using the seven steps of the Interpretative Phenomenological Analysis data analysis method. Emerging Findings Analysis to date has identified pertinent themes: • Difficulties of meaning-making for nurses because of the complexity of their boundary spanning role. • Emotional burden experienced in a highly emotive and fast-changing environment. • Stress and difficulties with role identity impacting on individual nurses’ ability to be resilient. • Challenges to wellbeing related to a sense of isolation when making complex decisions. Conclusion Emerging findings have highlighted the lived experiences of nurses working in liaison and diversion as challenging. The nature of the custody environment has an impact on role identity and decision making. Nurses left feeling isolated and unsupported are less resilient and may go on to experience compassion fatigue. The findings from this study thus far point to a need to connect nurses working in these boundary spanning roles with a supportive infrastructure where the complexity of their role is acknowledged, and they can be connected with a health agenda. In doing this, the nurses would be protected from harm and the likelihood of sustained positive outcomes for service users is optimised.

Keywords: liaison and diversion, nurse experiences, offender health, staff wellbeing

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Authors: Su Mon Saw, Joe Rothnagel

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Short open reading frames (sORFs) located in 5’UTR of mRNAs are known as uORFs. Characterization of uORF-encoded peptides (uPEPs) i.e., a subset of short open reading frame encoded peptides (sPEPs) and their translation regulation lead to understanding of causes of genetic disease, proteome complexity and development of treatments. Existence of uORFs within cellular proteome could be detected by LC-MS/MS. The ability of uORF to be translated into uPEP and achievement of uPEP identification will allow uPEP’s characterization, structures, functions, subcellular localization, evolutionary maintenance (conservation in human and other species) and abundance in cells. It is hypothesized that a subset of sORFs are translatable and that their encoded sPEPs are functional and are endogenously expressed contributing to the eukaryotic cellular proteome complexity. This project aimed to investigate whether sORFs encode functional peptides. Liquid chromatography-mass spectrometry (LC-MS) and bioinformatics were thus employed. Due to probable low abundance of sPEPs and small in sizes, the need for efficient peptide enrichment strategies for enriching small proteins and depleting the sub-proteome of large and abundant proteins is crucial for identifying sPEPs. Low molecular weight proteins were extracted using SDS-PAGE from Human Embryonic Kidney (HEK293) cells and Strong Cation Exchange Chromatography (SCX) from secreted HEK293 cells. Extracted proteins were digested by trypsin to peptides, which were detected by LC-MS/MS. The MS/MS data obtained was searched against Swiss-Prot using MASCOT version 2.4 to filter out known proteins, and all unmatched spectra were re-searched against human RefSeq database. ProteinPilot v5.0.1 was used to identify sPEPs by searching against human RefSeq, Vanderperre and Human Alternative Open Reading Frame (HaltORF) databases. Potential sPEPs were analyzed by bioinformatics. Since SDS PAGE electrophoresis could not separate proteins <20kDa, this could not identify sPEPs. All MASCOT-identified peptide fragments were parts of main open reading frame (mORF) by ORF Finder search and blastp search. No sPEP was detected and existence of sPEPs could not be identified in this study. 13 translated sORFs in HEK293 cells by mass spectrometry in previous studies were characterized by bioinformatics. Identified sPEPs from previous studies were <100 amino acids and <15 kDa. Bioinformatics results showed that sORFs are translated to sPEPs and contribute to proteome complexity. uPEP translated from uORF of SLC35A4 was strongly conserved in human and mouse while uPEP translated from uORF of MKKS was strongly conserved in human and Rhesus monkey. Cross-species conserved uORFs in association with protein translation strongly suggest evolutionary maintenance of coding sequence and indicate probable functional expression of peptides encoded within these uORFs. Translation of sORFs was confirmed by mass spectrometry and sPEPs were characterized with bioinformatics.

Keywords: bioinformatics, HEK293 cells, liquid chromatography-mass spectrometry, ProteinPilot, Strong Cation Exchange Chromatography, SDS-PAGE, sPEPs

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