Search results for: protein synthesis

Authors: Caroline Rocco, Feyza Karasu, Céline Croutxé-Barghorn, Xavier Allonas, Maxime Lecompère, Gérard Riess, Yujing Zhang, Catarina Esteves, Leendert van der Ven, Rolf van Benthem Gijsbertus de With

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Acrylates are widely used in UV-curing technology. Their high reactivity can, however, limit their conversion due to early vitrification. In addition, the free radical photopolymerization is known to be sensitive to oxygen inhibition leading to tacky surfaces. Although epoxides can lead to full polymerization, they are sensitive to humidity and exhibit low polymerization rate. To overcome the intrinsic limitations of both classes of monomers, Interpenetrating Polymer Networks (IPNs) can be synthesized. They consist of at least two cross linked polymers which are permanently entangled. They can be achieved under thermal and/or light induced polymerization in one or two steps approach. IPNs can display homogeneous to heterogeneous morphologies with various degrees of phase separation strongly linked to the monomer miscibility and also synthesis parameters. In this presentation, we synthesize UV-cured methacrylate - epoxide based IPNs with different chemical compositions in order to get a better understanding of their formation and phase separation. Miscibility before and during the photopolymerization, reaction kinetics, as well as mechanical properties and morphology have been investigated. The key parameters controlling the morphology and the phase separation, namely monomer miscibility and synthesis parameters have been identified. By monitoring the stiffness changes on the film surface, atomic force acoustic microscopy (AFAM) gave, in conjunction with polymerization kinetic profiles and thermomechanical properties, explanations and corroborated the miscibility predictions. When varying the methacrylate / epoxide ratio, it was possible to move from a miscible and highly-interpenetrated IPN to a totally immiscible and phase-separated one.

Keywords: investigation of properties and morphology, kinetics, phase separation, UV-cured IPNs

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Authors: Ekta Yadav, Sukanta Bhattacharya, Brijesh Yadav, Ariel Hus, Jagjit Yadav

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Background and Significance: Respiratory exposure to toxicants (chemicals or particulates) causes disruption of lung homeostasis leading to lung toxicity/injury manifested as pulmonary inflammation, edema, and/or other effects depending on the type and extent of exposure. This emphasizes the need for investigating toxicant type-specific mechanisms to understand therapeutic targets. Aquaporins, aka water channels, are known to play a role in lung homeostasis. Particularly, the two major lung aquaporins AQP5 and AQP1 expressed in alveolar epithelial and vasculature endothelia respectively allow for movement of the fluid between the alveolar air space and the associated vasculature. In view of this, the current study is focused on understanding the regulation of lung aquaporins and other targets during inhalation exposure to toxic chemicals (Cigarette smoke chemicals) versus toxic particles (Carbon nanoparticles) or co-exposures to understand their relevance as markers of injury and intervention. Methodologies: C57BL/6 mice (5-7 weeks old) were used in this study following an approved protocol by the University of Cincinnati Institutional Animal Care and Use Committee (IACUC). The mice were exposed via oropharyngeal aspiration to multiwall carbon nanotube (MWCNT) particles suspension once (33 ugs/mouse) followed by housing for four weeks or to Cigarette smoke Extract (CSE) using a daily dose of 30µl/mouse for four weeks, or to co-exposure using the combined regime. Control groups received vehicles following the same dosing schedule. Lung toxicity/injury was assessed in terms of homeostasis changes in the lung tissue and lumen. Exposed lungs were analyzed for transcriptional expression of specific targets (AQPs, surfactant protein A, Mucin 5b) in relation to tissue homeostasis. Total RNA from lungs extracted using TRIreagent kit was analyzed using qRT-PCR based on gene-specific primers. Total protein in bronchoalveolar lavage (BAL) fluid was determined by the DC protein estimation kit (BioRad). GraphPad Prism 5.0 (La Jolla, CA, USA) was used for all analyses. Major findings: CNT exposure alone or as co-exposure with CSE increased the total protein content in the BAL fluid (lung lumen rinse), implying compromised membrane integrity and cellular infiltration in the lung alveoli. In contrast, CSE showed no significant effect. AQP1, required for water transport across membranes of endothelial cells in lungs, was significantly upregulated in CNT exposure but downregulated in CSE exposure and showed an intermediate level of expression for the co-exposure group. Both CNT and CSE exposures had significant downregulating effects on Muc5b, and SP-A expression and the co-exposure showed either no significant effect (Muc5b) or significant downregulating effect (SP-A), suggesting an increased propensity for infection in the exposed lungs. Conclusions: The current study based on the lung toxicity mouse model showed that both toxicant types, particles (CNT) versus chemicals (CSE), cause similar downregulation of lung innate defense targets (SP-A, Muc5b) and mostly a summative effect when presented as co-exposure. However, the two toxicant types show differential induction of aquaporin-1 coinciding with the corresponding differential damage to alveolar integrity (vascular permeability). Interestingly, this implies the potential of AQP1 as a differential marker of toxicant type-specific lung injury.

Keywords: aquaporin, gene expression, lung injury, toxicant exposure

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Authors: Salma A. El-Marasy, Reham M. Abd-Elsalam, Omar A. Ahmed-Farid

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Dementia is characterized by impairments in memory and other cognitive abilities. This study aims to elucidate the possible ameliorative effect of silymarin on scopolamine-induced dementia using the object recognition test (ORT). The study was extended to demonstrate the role of cholinergic activity, oxidative stress, neuroinflammation, brain neurotransmitters and histopathological changes in the anti-amnestic effect of silymarin in demented rats. Wistar rats were pretreated with silymarin (200, 400, 800 mg/kg) or donepezil (10 mg/kg) orally for 14 consecutive days. Dementia was induced after the last drug administration by a single intraperitoneal dose of scopolamine (16 mg/kg). Then behavioral, biochemical, histopathological, and immunohistochemical analyses were then performed. Rats pretreated with silymarin counteracted scopolamine-induced non-spatial working memory impairment in the ORT and decreased acetylcholinesterase (AChE) activity, reduced malondialdehyde (MDA), elevated reduced glutathione (GSH), restored gamma-aminobutyric acid (GABA) and dopamine (DA) contents in the cortical and hippocampal brain homogenates. Silymarin dose-dependently reversed scopolamine-induced histopathological changes. Immunohistochemical analysis showed that silymarin dose-dependently mitigated protein expression of a glial fibrillary acidic protein (GFAP) and nuclear factor kappa-B (NF-κB) in the brain cortex and hippocampus. All these effects of silymarin were similar to that of the standard anti-amnestic drug, donepezil. This study reveals that the ameliorative effect of silymarin on scopolamine-induced dementia in rats using the ORT maybe in part mediated by, enhancement of cholinergic activity, anti-oxidant and anti-inflammatory activities as well as mitigation in brain neurotransmitters and histopathological changes.

Keywords: dementia, donepezil, object recognition test, rats, silymarin, scopolamine

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Authors: Sidra Shaw, Sarenna Shaw, Chae Joon Lee, Irimpan Mathews, Eric Koehn

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One of the biggest public health concerns of our time is increasing antimicrobial resistance. As of 2019, the CDC has documented more than 2.8 million serious antibiotic resistant infections in the United States. Currently, antibiotic resistant infections are directly implicated in over 750,000 deaths per year globally. On our current trajectory, British economist Jim O’Neill predicts that by 2050, an additional 10 million people (about half the population of New York) will die annually due to drug resistant infections. As a result, new biochemical pathways must be targeted to generate next generation antibiotic drugs that will be effective against drug resistant bacteria. One enticing target is the biosynthesis of DNA within bacteria, as few drugs interrupt this essential life process. Thymidylate synthase enzymes are essential for life as they catalyze the synthesis of a DNA building block, 2′-deoxythymidine-5′-monophosphate (dTMP). In humans, the thymidylate synthase enzyme (TSase) has been shown to be distinct from the flavin-dependent thymidylate synthase (FDTS) produced by many pathogenic bacteria. TSase and FDTS have distinct structures and mechanisms of catalysis, which should allow selective inhibition of FDTS over human TSase. Currently, C. diff is one of the most antibiotic resistant bacteria, and no drugs that target thymine biosynthesis exist for C. diff. Here we present the initial biochemical characterization of FDTS from C. diff. Specifically, we examine enzyme kinetics and binding features of this enzyme to determine the nature of interaction with ligands/inhibitors and understand the molecular mechanism of catalysis. This research will provide more insight into the targetability of the C. diff FDTS enzyme for novel antibiotic drugs.

Keywords: flavin-dependent thymidylate synthase, FDTS, clostridioides difficile, C. diff, antibiotic resistance, DNA synthesis, enzyme kinetics, binding features

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Authors: Yu-Chieh Chen, Li-Yun Wang, Chi-Chih Chen, Huy Hùng Đào, Ya-Mei Chen, Ming-Chu Cheng, Wen-Bin Chung, Hso-Chi Chaung, Guan-Ming Ke

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Many recent researches have demonstrated that CD154, a protein primarily expressed on activated T cell molecules, has potentially acted as a molecular adjuvant to improve the immunogenicity of subunit vaccines against viral infections. Classical swine fever (CSF) affects the swine industry worldwide that is one of the most devastating and highly contagious pig diseases. It is listed by the World Organization for Animal Health (OIE) as an infectious animal disease that must be reported. Although pigs vaccinated with subunit vaccines can be differentially diagnosed from those infected animals, subunit vaccines usually need adjuvants to enhance and elicit immune responses. In this study, CD154 was linked with CSFV E2 sequences and then expressed in CHO cells to produce the fusion protein as E2-CD154. The porcine specific CpG adjuvant was also used in one of the formulations. The specific pathogen-free pigs (SPF) at the age of 4-week-old were randomly separated into four groups, vaccinated with E2-CpG, E2-CD154, E2-CD154-CpG or the commercial Bayovac® CSF-E2 vaccine and boosted two weeks after primary vaccination. The results showed that the percentages of CD4+ and CD4+IL2+ in peripheral blood mononuclear cells (PBMC) in E2-CD154 vaccinated piglets seven days after primary vaccination were gained by 1-5% relative to the control group. In addition, the percentages of CD4+IFNγ+ T cells had slightly edged up 0.1-0.3% compared with the control group. Also, increased E2-specific IFNγ levels had edged up CD4+CD8+ T cells found in E2-CD154 and E2-CD154-CpG groups, particularly in the E2-CD154-CpG group. These results implicate that CD154 may enhance cellular immunity and synergistically act with species-specific CpG adjuvant as a dual-phase adjuvant. Therefore, the CD154 may be beneficial as a promising adjuvant in subunit vaccines.

Keywords: CD154, CpG adjuvant, cellular immunity, subunit vaccine, pig

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Authors: K. Shailaja

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The advanced glycation end products (AGEs) formed between the reducing sugar and protein as a result of Oxidative stress and non-enzymatic glycosylation play an important role in pathogenesis of diabetes and aging complication. Glycation results in the production of free radicals. The oxidation process is believed to play an important role in AGEs formation. Thus agents with antioxidative property and antiglycation activity may retard the process of AGEs formation. Selected medicinal plants for the present study include Catharanthus roseus, Bougainvillea spectabilis (pink flowers), Cinnamomum tamala, Cinnamomum zeylanica, Abutilon indicum, Asparagus racemosus, and Sapindus emarginatus. The crude ethanolic extracts of the selected medicinal plants at varying concentrations ranging from 1-100 mg/ml were evaluated for in vitro antioxidant and protein glycation activities by FRAP and glucose-BSA assay respectively. Among all the plants tested, Bougainvillea spectabilis, Catharanthus roseus and Abutilon indicum showed strong antioxidant activity The antioxidant activity was expressed as mg of Gallic acid/ gm sample which was found to be 4.3 mg, 1.3mg, and 1.3mg respectively for Bougainvillea spectabilis, Catharanthus roseus and Abutilon indicum. The results of inhibition of the initial glycation product i.e., fructosamine was found to be 35% for Asparagus racemosus, Cinnamomum tamala and Abutilon indicum followed by the other plant extracts. The results indicate that these plants are potential sources of natural antioxidants which have free radical scavenging activity and might be used not only for reducing oxidative stress in diabetes but also open a new research avenues in the field of Natural Products.

Keywords: in vitro antioxidant activity, anti-glycation activity, ethanol extracts, polyphenols, Catharanthus roseus, Cinnamomum tamala

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Authors: Ting Pan, Jiacheng Wang, Pengcheng Lin, Ying Chen, Songping Mo

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Phase change materials (PCMs) are widely used in latent heat thermal energy storage because of their good properties such as high energy storage density and constant heat-storage/release temperature. Microencapsulation techniques can prevent PCMs from leaking during the liquid-solid phase transition and enhance thermal properties. This technique has been widely applied in architectural materials, thermo-regulated textiles, aerospace fields, etc. One of the most important processes during the synthesis of microcapsules is to form a stable emulsion of the PCM core and reactant solution for the formation of the shell of the microcapsules. The use of surfactants is usually necessary for the formation of a stable emulsion system because of the difference in hydrophilia/lipophilicity of the PCM and the solvent. Unfortunately, the use of surfactants may cause pollution to the environment. In this study, modified kaolinite was used as an emulsion stabilizer for the microencapsulation of octodecane as PCM. Microcapsules were synthesized by phase inversion emulsification method, and the shell of polymethyl methacrylate (PMMA) was formed through free radical polymerization. The morphologies, crystalloid phase, and crystallization properties of microcapsules were investigated using scanning electron microscopy (SEM), X-ray diffractometer (XRD), and Fourier transforms infrared spectrometer (FTIR). The thermal properties and thermal stability were investigated by a differential scanning calorimeter (DSC) and a thermogravimetric analyzer (TG). The FT-IR, XRD results showed that the octodecane was well encapsulated in the PMMA shell. The SEM results showed that the microcapsules were spheres with an average size of about 50-100nm. The DSC results indicated that the latent heat of the microcapsules was 152.64kJ/kg and 164.23kJ/kg. The TG results confirmed that the microcapsules had good thermal stability due to the PMMA shell. Based on the results, it can be concluded that the modified kaolinite can be used as an emulsifier for the synthesis of PCM microcapsules, which is valid for reducing part of the possible pollution caused by the utilization of surfactants.

Keywords: kaolinite, microencapsulation, PCM, thermal energy storage

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Authors: Z. Karahaliloğlu, E. Yalçın, M. Demirbilek, E.B. Denkbaş

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Critical-sized bone defects caused by trauma, bone diseases, prosthetic implant revision or tumor excision cannot be repaired by physiological regenerative processes. Current orthopedic applications for critical-sized bone defects are to use autologous bone grafts, bone allografts, or synthetic graft materials. However, these strategies are unable to solve completely the problem, and motivate the development of novel effective biological scaffolds for tissue engineering applications and regenerative medicine applications. In particular, scaffolds combined with a variety of bio-agents as fundamental tools emerge to provide the regeneration of damaged bone tissues due to their ability to promote cell growth and function. In this study, a magnetic silk fibroin (SF) hydrogel scaffold was prepared by electrogelation process of the concentrated Bombxy mori silk fibroin (8 %wt) aqueous solution. For enhancement of osteoblast-like cells (SaOS-2) growth and adhesion, basal fibroblast growth factor (bFGF) were conjugated physically to the HSA-coated magnetic nanoparticles (Fe3O4) and magnetic SF e-gel scaffolds were prepared by incorporation of Fe3O4, HSA (human serum albumin)=Fe3O4 and HSA=Fe3O4-bFGF nanoparticles. HSA=Fe3O4, HSA=Fe3O4-bFGF loaded and bare SF e-gels scaffolds were characterized using scanning electron microscopy (SEM.) For cell studies, human osteoblast-like cell line (SaOS-2) was used and an MTT assay was used to assess the cytotoxicity of magnetic silk fibroin e-gel scaffolds and cell density on these surfaces. For the evaluation osteogenic activation, ALP (alkaline phosphatase), the amount of mineralized calcium, total protein and collagen were studied. Fe3O4 nanoparticles were successfully synthesized and bFGF was conjugated to HSA=Fe3O4 nanoparticles with %97.5 of binding yield which has a particle size of 71.52±2.3 nm. Electron microscopy images of the prepared HSA and bFGF incorporated SF e-gel scaffolds showed a 3D porous morphology. In terms of water uptake results, bFGF conjugated HSA=Fe3O4 nanoparticles has the best water absorbability behavior among all groups. In the in-vitro cell culture studies realized using SaOS-2 cell line, the coating of Fe3O4 nanoparticles surface with a protein enhance the cell viability and HSA coating and bFGF conjugation, the both have an inductive effect in the cell proliferation. One of the markers of bone formation and osteoblast differentiation, according to the ALP activity and total protein results, HSA=Fe3O4-bFGF loaded SF e-gels had significantly enhanced ALP activity. Osteoblast cultured HSA=Fe3O4-bFGF loaded SF e-gels deposited more calcium compared with SF e-gel. The proposed magnetic scaffolds seem to be promising for bone tissue regeneration and used in future work for various applications.

Keywords: basic fibroblast growth factor (bFGF), e-gel, iron oxide nanoparticles, silk fibroin

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Authors: Maher J. Tadros

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This research was conducted to study the effect of different salinity concentrations on the plant water relation and forage quality on two multipurpose forest trees species seedlings Leucaena leucocephala (Lam.) de wit and Acacia saligna (Labill.). Five different salinity concentrations mixture between sodium chloride and calcium chloride (v/v, 1:1) were applied. The control (Distilled Water), 2000, 4000, 6000, and 8000 ppm were used to water the seedlings for 3 months. The research results presented showed a marked variation among the two species in response to salinity. The Leucaena was able to withstand the highest level of salinity compared to Acacia all over the studied parameters except in the relative water content. Although all the morphological characteristics studied for the two species showed a marked decrease under the different salinity concentrations, except the shoot/root ratio that showed a trend of increase. The water stress measure the leaf water potential was more negative with as the relative water content increase under that saline conditions compared to the control. The forage quality represented by the crude protein and nitrogen content were low at 6000 ppm compared to the 8000 ppm in L. Leucocephala that increased compared that level in A. saligna. Also the results showed that growing both Leucaena and Acacia provide a good source of forage when that grow under saline condition which will be of great benefits to the agricultural sector especially in the arid and semiarid areas were these species can provide forage with high quality forage all year around when grown under irrigation with saline. This research recommended such species to be utilized and grown for forages under saline conditions.

Keywords: plant water relations, growth performance, salinity stress, protein content, forage quality, multipurpose trees

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Authors: Cristian Moisa, Andreea Lupitu, Adriana Csakvari, Dana G. Radu, Leonard Marian Olariu, Georgeta Pop, Dorina Chambre, Lucian Copolovici, Dana Copolovici

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Green synthesis of nanoparticles (NPs) represents a promising, accessible, eco-friendly, and safe process with significant applications in biotechnology, pharmaceutical sciences, and farming. The aim of our study was to obtain silver nanoparticles, using plant wastes extracts resulted in the essential oils extraction process: Thymus vulgaris L., Origanum vulgare L., Lavandula angustifolia L., and in hemp processing for seed and fibre, Cannabis sativa. Firstly, we obtained aqueous extracts of thyme, oregano, lavender, and hemp (two monoicous and one dioicous varieties), all harvested in western part of Romania. Then, we determined the chemical composition of the extracts by liquid-chromatography coupled with diode array and mass spectrometer detectors. The compounds identified in the extracts were in agreement with earlier published data, and the determination of the antioxidant activity of the obtained extracts by DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) assays confirmed their antioxidant activity due to their total polyphenolic content evaluated by Folin-Ciocalteu assay. Then, the silver nanoparticles (AgNPs) were successfully biosynthesised, as was demonstrated by UV-VIS, FT-IR spectroscopies, and SEM, by reacting AgNO₃ solution and plant extracts. AgNPs were spherical in shape, with less than 30 nm in diameter, and had a good bactericidal activity against Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli, Klebsiella pneumoniae, Pseudomonas fluorescens).

Keywords: plant wastes extracts, chemical composition, high performance liquid chromatography mass spectrometer, HPLC-MS, scanning electron microscopy, SEM, silver nanoparticles

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Authors: M. Naimi, M. B. Khaled

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The present study consisted of an applied test in meat system to assess the effectiveness of three bio agents bacteriocinproducing strains: Lm24: Lactobacillus sakei, Lm14and Lm25: Pediococcus spp. Two tests were carried out: The ex-situ test was intended for three batches added with crude bacteriocin solutions at 12.48 AU/ml for Lm25 and 8.4 AU/ml for Lm14 and Lm24. However, the in situ one consisted of four batches; three of them inoculated with one bacteriocinogenic Lm25, Lm14, Lm24, respectively. The fourth one was used in mixture: Lm14+m24 at approximately of 107 CFU/ml. The two used tests were done in the presence of the pathogen St. aureus ATCC 6538, as a test strain at 103 CFU/ml. Another batch served as a positive or a negative control was used too. The incubation was performed at 7°C. Total viable counts, staphylococci and lactic acid bacteria, at the beginning and at selected times with interval of three days were enumerated. Physicochemical determinations (except for in situ test): pH, dry mater, sugars, fat and total protein, at the beginning and at end of the experiment, were done, according to the international norms. Our results confirmed the ex situ effectiveness. Furthermore, the batches affected negatively the total microbial load over the incubation days, and showed a significant regression in staphylococcal load at day seven, for Lm14, Lm24, and Lm25 of 0.73, 2.11, and 2.4 log units. It should be noticed that, at the last day of culture, staphylococcal load was nil for the three batches. In the in situ test, the cultures displayed less inhibitory attitude and recorded a decrease in staphylococcal load, for Lm14, Lm24, Lm25, Lm14+m24 of 0.73, 0.20, 0.86, 0.032 log units. Therefore, physicochemical analysis for Lm14, Lm24, Lm25, Lm14+m24 showed an increase in pH from 5.50 to 5.77, 6.18, 5.96, 7.22, a decrease in dry mater from 7.30% to 7.05%, 6.87%, 6.32%, 6.00%.This result reflects the decrease in fat ranging from 1.53% to 1.49%, 1.07%, 0.99%, 0.87%; and total protein from 6.18% to 5.25%, 5.56%, 5.37%, 5.5%. This study suggests that the use of selected strains as Lm25 could lead to the best results and would help in preserving and extending the shelf life of lamb meat.

Keywords: biocontrol, in situ, ex situ, meat system, St. aureus, Lactobacillus sakei, Pediococcus spp.

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Authors: Insaf Mehani, Bachir Bouchekima

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The fight against climate change and the replacement of fossil energies nearing exhaustion gradually emerge as major societal and economic challenges. It is possible to develop common dates of low commercial value, and put on the local and international market a new generation of products with high added values such as bio ethanol. Besides its use in chemical synthesis, bio ethanol can be blended with gasoline to produce a clean fuel while improving the octane.

Keywords: bio-energy, waste dates, bio ethanol, Algeria

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Authors: N. Umasuthan, S. D. N. K. Bathige, W. S. Thulasitha, I. Whang, J. Lee

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The MyD88 is an evolutionarily conserved host-expressed adaptor protein that is essential for proper TLR/ IL1R immune-response signaling. A previously identified complete cDNA (1626 bp) of OfMyD88 comprised an ORF of 867 bp encoding a protein of 288 amino acids (32.9 kDa). The gDNA (3761 bp) of OfMyD88 revealed a quinquepartite genome organization composed of 5 exons (with the sizes of 310, 132, 178, 92 and 155 bp) separated by 4 introns. All the introns displayed splice signals consistent with the consensus GT/AG rule. A bipartite domain structure with two domains namely death domain (24-103) coded by 1st exon, and TIR domain (151-288) coded by last 3 exons were identified through in silico analysis. Moreover, homology modeling of these two domains revealed a similar quaternary folding nature between human and rock bream homologs. A comprehensive comparison of vertebrate MyD88 genes showed that they possess a 5-exonic structure. In this structure, the last three exons were strongly conserved, and this suggests that a rigid structure has been maintained during vertebrate evolution. A cluster of TATA box-like sequences were found 0.25 kb upstream of cDNA starting position. In addition, putative 5'-flanking region of OfMyD88 was predicted to have TFBS implicated with TLR signaling, including copies of NFB1, APRF/ STAT3, Sp1, IRF1 and 2 and Stat1/2. Using qPCR technique, a ubiquitous mRNA expression was detected in liver and blood. Furthermore, a significantly up-regulated transcriptional expression of OfMyD88 was detected in head kidney (12-24 h; >2-fold), spleen (6 h; 1.5-fold), liver (3 h; 1.9-fold) and intestine (24 h; ~2-fold) post-Fla challenge. These data suggest a crucial role for MyD88 in antibacterial immunity of teleosts.

Keywords: MyD88, innate immunity, flagellin, genomic analysis

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Authors: Sulalit Bandyopadhyay, Muhammad Awais Ashfaq Alvi, Anuvansh Sharma, Wilhelm R. Glomm

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Release of drugs from nanogels and nanogel-based systems can occur under the influence of external stimuli like temperature, pH, magnetic fields and so on. pNIPAm-AAc nanogels respond to the combined action of both temperature and pH, the former being mostly determined by hydrophilic-to-hydrophobic transitions above the volume phase transition temperature (VPTT), while the latter is controlled by the degree of protonation of the carboxylic acid groups. These nanogels based systems are promising candidates in the field of drug delivery. Combining nanogels with magneto-plasmonic nanoparticles (NPs) introduce imaging and targeting modalities along with stimuli-response in one hybrid system, thereby incorporating multifunctionality. Fe@Au core-shell NPs possess optical signature in the visible spectrum owing to localized surface plasmon resonance (LSPR) of the Au shell, and superparamagnetic properties stemming from the Fe core. Although there exist several synthesis methods to control the size and physico-chemical properties of pNIPAm-AAc nanogels, yet, there is no comprehensive study that highlights the dependence of incorporation of one or more layers of NPs to these nanogels. In addition, effective determination of volume phase transition temperature (VPTT) of the nanogels is a challenge which complicates their uses in biological applications. Here, we have modified the swelling-collapse properties of pNIPAm-AAc nanogels, by combining with Fe@Au NPs using different solution based methods. The hydrophilic-hydrophobic transition of the nanogels above the VPTT has been confirmed to be reversible. Further, an analytical method has been developed to deduce the average VPTT which is found to be 37.3°C for the nanogels and 39.3°C for nanogel coated Fe@Au NPs. An opposite swelling –collapse behaviour is observed for the latter where the Fe@Au NPs act as bridge molecules pulling together the gelling units. Thereafter, Cyt C, a model protein drug and L-Dopa, a drug used in the clinical treatment of Parkinson’s disease were loaded separately into the nanogels and nanogel coated Fe@Au NPs, using a modified breathing-in mechanism. This gave high loading and encapsulation efficiencies (L Dopa: ~9% and 70µg/mg of nanogels, Cyt C: ~30% and 10µg/mg of nanogels respectively for both the drugs. The release kinetics of L-Dopa, monitored using UV-vis spectrophotometry was observed to be rather slow (over several hours) with highest release happening under a combination of high temperature (above VPTT) and acidic conditions. However, the release of L-Dopa from nanogel coated Fe@Au NPs was the fastest, accounting for release of almost 87% of the initially loaded drug in ~30 hours. The chemical structure of the drug, drug incorporation method, location of the drug and presence of Fe@Au NPs largely alter the drug release mechanism and the kinetics of these nanogels and Fe@Au NPs coated with nanogels.

Keywords: controlled release, nanogels, volume phase transition temperature, l-dopa

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Authors: Deniz Dincel, Sena Ardicli, Hale Samli, Mustafa Ogan, Faruk Balci

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The aim of the study was to determine the effects of some environmental and genetic factors on growth, fertility traits, milk yield and composition in Saanen goats. For this purpose, the total of 173 Saanen goats and kids were investigated for growth, fertility and milk traits in Marmara Region of Turkey. Fertility parameters (n=70) were evaluated during two years. Milk samples were collected during the lactation and the milk yield/components (n=59) of each goat were calculated. In terms of CSN3 and AGPAT6 gene; the genotypes were defined by PCR-RFLP. Saanen kids (n=86-112) were measured from birth to 6 months of life. The birth, weaning, 60ᵗʰ, 90ᵗʰ, 120ᵗʰ and 180tᵗʰ days of average live weights were calculated. The effects of maternal age on pregnancy rate (p < 0.05), birth rate (p < 0.05), infertility rate (p < 0.05), single born kidding (p < 0.001), twinning rate (p < 0.05), triplet rate (p < 0.05), survival rate of kids until weaning (p < 0.05), number of kids per parturition (p < 0.01) and number of kids per mating (p < 0.01) were found significant. The impacts of year on birth rate (p < 0.05), abortion rate (p < 0.001), single born kidding (p < 0.01), survival rate of kids until weaning (p < 0.01), number of kids per mating (p < 0.01) were found significant for fertility traits. The impacts of lactation length on all milk yield parameters (lactation milk, protein, fat, totally solid, solid not fat, casein and lactose yield) (p < 0.001) were found significant. The effects of age on all milk yield parameters (lactation milk, protein, fat, total solid, solid not fat, casein and lactose yield) (p < 0.001), protein rate (p < 0.05), fat rate (p < 0.05), total solid rate (p < 0.01), solid not fat rate (p < 0.05), casein rate (p < 0.05) and lactation length (p < 0.01), were found significant too. However, the effect of AGPAT6 gene on milk yield and composition was not found significant in Saanen goats. The herd was found monomorphic (FF) for CSN3 gene. The effects of sex on live weights until 90ᵗʰ days of life (birth, weaning and 60ᵗʰ day of average weight) were found significant statistically (p < 0.001). The maternal age affected only birth weight (p < 0,001). The effects month at birth on all of the investigated day [the birth, 120ᵗʰ, 180ᵗʰ days (p < 0.05); the weaning, 60ᵗʰ, 90ᵗʰ days (p < 0,001)] were found significant. The birth type was found significant on the birth (p < 0,001), weaning (p < 0,01), 60ᵗʰ (p < 0,01) and 90ᵗʰ (p < 0,01) days of average live weights. As a result, screening the other regions of CSN3, AGPAT6 gene and also investigation the phenotypic association of them should be useful to clarify the efficiency of target genes. Environmental factors such as maternal age, year, sex and birth type were found significant on some growth, fertility and milk traits in Saanen goats. So consideration of these factors could be used as selection criteria in dairy goat breeding.

Keywords: fertility, growth, milk yield, Saanen goats

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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

Procedia PDF Downloads 108

Authors: Widari

Abstract:

This study aims to determine the relationship of fast food consumption preferences with macro and micro nutrient adequacy students of SMP Negeri 5 Padang. This study used a cross sectional study conducted on 100 students of SMP Negeri 5 Padang. The variables studied were fast food preferences, nutrition adequacy macronutrients (carbohydrate, protein, fat, fiber) and micro nutrients (sodium, calcium, iron). Confounding factor in this study was the physical activity level because it was considered quite affecting food consumption of students. Data collected by using a questionnaire food recall as many as 2 x 24 hours to see the history of the respondents eat at school day and on holidays. Then, data processed using software Nutrisurvey and Microsoft Excel 2010. The analysis was performed on samples that have low and medium category on physical activity. The physical activity was not analyzed with another variable to see the strength of the relationship between independent and dependent variables. So that, do restrictions on physical activity variables in an attempt to get rid of confounding in design. Univariate and bivariate analyzes performed using SPSS 16.0 for Windows with Kolmogrov-Smirnov statistical tests, confidence level = 95% (α = 0,05). Results of univariate analysis showed that more than 70% of respondents liked fast food. On average, respondents were malnourished macro; malnourished fiber (100%), carbohydrates (72%), and protein (56%), whereas for fat, excess intake of the respondents (41%). Furthermor, many respondents who have micronutrient deficiencies; 98% for sodium, 96% for iron, and 91% for calcium. The results of the bivariate analysis showed no significant association between fast food consumption preferences with macro and micro nutrient adequacy (p > 0,05). This happens because in the fact not all students who have a preference for fast food actually eat them. To study better in the future, it is expected sampling really like and eat fast food in order to obtain better analysis results.

Keywords: fast food, nutritional adequacy, preferences, students

Procedia PDF Downloads 368

Authors: J. Ritonja, B. Grcar

Abstract:

For the synchronous generator simulation and analysis and for the power system stabilizer design and synthesis a mathematical model of synchronous generator is needed. The model has to accurately describe dynamics of oscillations, while at the same time has to be transparent enough for an analysis and sufficiently simplified for design of control system. To study the oscillations of the synchronous generator against to the rest of the power system, the model of the synchronous machine connected to an infinite bus through a transmission line having resistance and inductance is needed. In this paper, the linearized reduced order dynamic model of the synchronous generator connected to the infinite bus is presented and analysed in details. This model accurately describes dynamics of the synchronous generator only in a small vicinity of an equilibrium state. With the digression from the selected equilibrium point the accuracy of this model is decreasing considerably. In this paper, the equations’ descriptions and the parameters’ determinations for the linearized reduced order mathematical model of the synchronous generator are explained and summarized and represent the useful origin for works in the areas of synchronous generators’ dynamic behaviour analysis and synchronous generator’s control systems design and synthesis. The main contribution of this paper represents the detailed analysis of the accuracy of the linearized reduced order dynamic model in the entire synchronous generator’s operating range. Borders of the areas where the linearized reduced order mathematical model represents accurate description of the synchronous generator’s dynamics are determined with the systemic numerical analysis. The thorough eigenvalue analysis of the linearized models in the entire operating range is performed. In the paper, the parameters of the linearized reduced order dynamic model of the laboratory salient poles synchronous generator were determined and used for the analysis. The theoretical conclusions were confirmed with the agreement of experimental and simulation results.

Keywords: eigenvalue analysis, mathematical model, power system stability, synchronous generator

Procedia PDF Downloads 236

Authors: Pham Minh Duc, Tran Thi Thanh Hien, David A. Bengtson

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Laboratory feeding trial: the study was conducted to find out the optimal dietary vitamin C, or ascorbic acid (AA) levels in terms of the growth performance of snakehead. The growth trial included six treatments with five replications. Each treatment contained 0, 125, 250, 500, 1000 and 2000 mg AA equivalent kg⁻¹ diet which included six iso-nitrogenous (45% protein), iso-lipid (9% lipid) and isocaloric (4.2 Kcal.g¹). Eighty snakehead fingerlings (6.24 ± 0.17 g.fish¹) were assigned randomly in 0.5 m³ composite tanks. Fish were fed twice daily on demand for 8 weeks. The result showed that growth rates increased, protein efficiency ratio increased and the feed conversion ratio decreased in treatments with AA supplementation compared with control treatment. The survival rate of fish tends to increase with increase AA level. The number of RBCs, lysozyme in treatments with AA supplementation tended to rise significantly proportional to the concentration of AA. The number of WBCs of snakehead in treatments with AA supplementation was higher 2.1-3.6 times. In general, supplementation of AA in the diets for snakehead improved growth rate, feed efficiency and immune response. Hapa on-farm trial: based on the results of the laboratory feeding trial, the effects of AA on snakehead in hapas to simulate farm conditions, was tested using the following treatments: commercial feed; commercial feed plus hand mixed AA at 500; 750 and 1000 mg AA.kg⁻¹; SBM diet without AA; SBM diet plus 500; 750 and 1000 mg AA.kg⁻¹. The experiment was conducted in two experimental ponds (only SBM diet without AA placed in one pond and the rest in the other pond) with four replicate hapa each. Stocking density was 150 fish.m² and culture period was 5 months until market size was attained. The growth performance of snakehead and economic aspects were examined in this research.

Keywords: fish health, growth rate, snakehead, Vitamin C

Procedia PDF Downloads 93

Authors: Louise Elizabeth Bolton

Abstract:

Statement of the problem: There are an estimated 328 million cases of COPD worldwide. It is likely to become the third biggest cause of death by 2030. The impact of living with palliative care needs arising from COPD disrupts an individual’s existential situation. Understandings of individuals' existential situations within COPD are limited within the research literature and are rarely addressed within clinical practice, yet existential suffering has been linked to poor health-related quality of life for those living with other chronic conditions. The purpose of this integrative review is to provide a synthesis of existing evidence on existential suffering for those living with palliative care needs arising from COPD. Methods: This is an integrative review undertaken in accordance with PRISMA guidelines. Nine electronic databases were searched from April 2019 to January 2021. Thirty-five empirical research papers of both qualitative and quantitative methodologies, alongside systematic literature reviews, were included. Data analysis was undertaken using an integrative thematic analysis approach. Findings: Identified themes of existential suffering when living with palliative care needs arising from COPD are as follows: Liminality, Lamented Life, Loss of Personal Liberty, Life Meaning and Existential isolation. The absence of life meaning and purpose was of most importance to patients. Conclusion and Significance: This integrative review provides a synthesis of international evidence upon the presence of existential suffering. It is present and of significant impact within the daily lives of those living with palliative care needs arising from COPD. The absence of life meaning has the most significant impact, requiring further exploration of both its physical and psychological impact. Rediscovery of life meaning diminishes feelings of worthlessness and hopelessness in daily life and facilitates feelings of inner peace. For those with COPD living with such a relentless symptom burden, a positive existential situation is desirable.

Keywords: palliative care, COPD, existential suffering, end of life care

Procedia PDF Downloads 130

Authors: Hosia T. Pule, Julius T. Tjelele, Michelle J. Tedder, Dawood Hattas

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Acceptability of plant material to herbivores is influenced by, among other factors; nutrients, plant secondary metabolites and growth stage of the plants. However, the effect of these factors on Seriphium plumosum L. acceptability to livestock is still not clearly understood, despite its importance in managing its encroachment in grassland communities. The study used 2 x 2 x 2 factorial analysis of variance to investigate the effect of season (wet and dry), fire, slope position (top and bottom) and their interaction on Seriphium plumosum chemistry. We tested the hypothesis that S. plumosum chemistry varies temporally, spatially and pre- and post-fire treatment. Seriphium plumosum edible material was collected during the wet and dry season from burned and unburned areas on both top and bottom slopes before being analysed for protein (CP) content, neutral detergent fibre (NDF), total phenolics (TP) and condensed tannins (CT). Season had a significant effect on S. plumosum protein content, neutral detergent fibre, total phenolics and condensed tannins. Fire had a significant effect on CP. Interaction of season x fire had a significant effect on NDF and CP (p < 0.05). Seriphium plumosum in the wet season (6.69% ± 0.20 (SE)) had significantly higher CP than in the dry season (5.22% ± 0.13). NDF was significantly higher (58.01% ± 0.41) in the dry season than in the wet season (53.17% ± 0.34), while TP were significantly higher in the dry season (14.44 mg/gDw ± 1.03) than in the wet season (11.08 mg/gDw ± 1.07). CT in the wet season were significantly higher (1.56 mg/gDw ± 0.13) than in the dry season (1 mg/gDw ± 0.03). CP was significantly higher in burned (6. 31 % ± 0.22) than in unburned S. plumosum edible material (5.60 % ± 0.15). Seriphium plumosum CP was significantly higher in wet season x burned (7.34 % ± 0.31) than wet season x unburned (6.08 % ± 0.20) material and dry season x burned (5.34 % ± 0.18) and unburned (5.09 % ± 0.18) material were similar. NDF was similar in dry season x burned (58.31% ± 0.54) and dry season x unburned (57.69 % ± 0.62) material and significantly higher than similar wet season x burned (52.43% ± 0.45) and wet season x post-unburned (53.88% ± 0.47) material. This study suggests integrating fire, browsers, and supplements as encroacher S. plumosum control agents, especially in the wet season, following fire due to high S. plumosum CP content.

Keywords: acceptability, chemistry, edible material, encroachment, phenolics, tannins

Procedia PDF Downloads 150

Authors: Ali Kassem, Yhea Kishik, Ali Hassan, Mohamed Abdelwahab

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Introduction: Accelerated coronary and peripheral vascular atherosclerosis is one of the most common and chronic complications of diabetes mellitus. A recent aspect of coronary artery disease in this condition is its silent nature. The aim of the work: Detection of the prevalence of silent myocardial ischemia (SMI) in Upper Egypt type 2 diabetic males and to select male diabetic population who should be screened for SMI. Patients and methods: 100 type 2 diabetic male patients with a negative history of angina or anginal equivalent symptoms and 30 healthy control were included. Full medical history and thorough clinical examination were done for all participants. Fasting and post prandial blood glucose level, lipid profile, (HbA1c), microalbuminuria, and C-reactive protein were done for all participants Resting ECG, trans-thoracic echocardiography, treadmill exercise ECG, myocardial perfusion imaging were done for all participants and patients positive for one or more NITs were subjected for coronary angiography. Results Twenty nine patients (29%) were positive for one or more NITs in the patients group compared to only one case (3.3%) in the controls. After coronary angiography, 20 patients were positive for significant coronary artery stenosis in the patients group, while it was refused to be done by the patient in the controls. There were statistical significant difference between the two groups regarding, hypertension, dyslipidemia and obesity, family history of DM and IHD with higher levels of microalbuminuria, C-reactive protein, total lipids in patient group versus controls According to coronary angiography, patients were subdivided into two subgroups, 20 positive for SMI (positive for coronary angiography) and 80 negative for SMI (negative for coronary angiography). No statistical difference regarding family history of DM and type of diabetic therapy was found between the two subgroups. Yet, smoking, hypertension, obesity, dyslipidemia and family history of IHD were significantly higher in diabetics positive versus those negative for SMI. 90% of patients in subgroup positive for SMI had two or more cardiac risk factors while only two patients had one cardiac risk factor (10%). Uncontrolled DM was detected more in patients positive for SMI. Diabetic complications were more prevalent in patients positive for SMI versus those negative for SMI. Most of the patients positive for SMI have DM more than 5 years duration. Resting ECG and resting Echo detected only 6 and 11 cases, respectively, of the 20 positive cases in group positive for SMI compared to treadmill exercise ECG and myocardial perfusion imaging that detected 16 and 18 cases respectively, Conclusion: Type 2 diabetic male patients should be screened for detection of SMI when aged above 50 years old, diabetes duration is more than 5 years, presence of two or more cardiac risk factors and/or patients suffering from one or more of the chronic diabetic complications. CRP, is an important parameter for selection of type 2 diabetic male patients who should be screened for SMI. Non invasive cardiac tests are reliable for screening of SMI in these patients in our locality.

Keywords: C-reactive protein, Silent myocardial ischemia, Stress tests, type 2 DM

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Authors: Sneha Kumari, Ravi Krishnan Elangovan

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This abstract describes the development of a high-speed tracking method by modification in motor components for nanoparticle attachment. Myosin motors are nano-sized protein machines powering movement that defines life. These miniature molecular devices serve as engines utilizing chemical energy stored in ATP to produce useful mechanical energy in the form of a few nanometre displacement events leading to force generation that is required for cargo transport, cell division, cell locomotion, translated to macroscopic movements like running etc. With the advent of in vitro motility assay (IVMA), detailed functional studies of the actomyosin system could be performed. The major challenge with the currently available IVMA for tracking actin filaments is a resolution limitation of ± 50nm. To overcome this, we are trying to develop Single Molecule IVMA in which nanoparticle (GNP/QD) will be attached along or on the barbed end of actin filaments using CapZ protein and visualization by a compact TIRF module called ‘cTIRF’. The waveguide-based illumination by cTIRF offers a unique separation of excitation and collection optics, enabling imaging by scattering without emission filters. So, this technology is well equipped to perform tracking with high precision in temporal resolution of 2ms with significantly improved SNR by 100-fold as compared to conventional TIRF. Also, the nanoparticles (QD/GNP) attached to actin filament act as a point source of light coffering ease in filament tracking compared to conventional manual tracking. Moreover, the attachment of cargo (QD/GNP) to the thin filament paves the way for various nano-technological applications through their transportation to different predetermined locations on the chip

Keywords: actin, cargo, IVMA, myosin motors and single-molecule system

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Authors: Macdalyna Esther Ronie, Hasmadi Mamat, Ahmad Hazim Abdul Aziz, Mohamad Khairi Zainol

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Current trends in gluten-free food development are increasingly leaning towards the utilization of pigmented rice flour, with a particular focus on Bario Merah Sederhana (BMS), a red-pigmented rice native to Sarawak, Malaysia. This study delves into the evaluation of the nutritional, textural, and sensory attributes of gluten-free rice bread produced from a blend of BMS rice flour and potato starch. The resulting samples are denoted as F1 (100% BMS rice flour), F2 (90% BMS rice flour and 10% potato starch), F3 (80% BMS rice flour and 20% potato starch), and F4 (70% BMS rice flour and 30% potato starch). Comparatively, these gluten-free rice bread formulations exhibit higher levels of ash and crude fiber, along with lower carbohydrate content when juxtaposed with conventional wheat bread. Notably, the crude protein content of the rice bread diminishes significantly (p<0.05) as the proportion of rice flour decreases, primarily due to the higher protein content found in wheat flour. The crumb of the rice bread appears darker owing to the red pigment in the rice flour, while the crust is lighter than that of the control sample, possibly attributable to a reduced Maillard reaction. Among the various rice bread formulations, F4 stands out with the least dough and bread hardness, accompanied by the highest levels of stickiness and springiness in both dough and bread, respectively. In sensory evaluations, wheat bread garners the highest rating (p<0.05). However, within the realm of rice breads, F4 emerges as a viable and acceptable formulation, as indicated by its commendable scores in color (7.03), flavor (5.73), texture (6.03), and overall acceptability (6.18). These findings underscore the potential of BMS in the creation of gluten-free rice breads, with the formulation consisting of 70% rice flour and 30% potato starch emerging as a well-received and suitable option.

Keywords: gluten-free bread, bario rice, proximate composition, sensory evaluation

Procedia PDF Downloads 232

Authors: Sahar Javadi-Novashnagh, Mohammad Moradi-Shahrbabak, Mostafa Sadeghi, Katarzyna Ropka-Molik, Hossein Moradi-Shahrbabak, Maria Consuelo Mura

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The objective of this study was to investigate two single nucleotide polymorphisms located in exon 2 (g.939A > G) and intron 3 (g.4349A > G) of fatty acid binding protein 3 (FABP3) gene in two Iranian sheep breeds, Lori-Bakhtiari and Zel, using polymerase chain reaction -restriction fragment length polymorphism (PCR-RFLP) approach. The association of the polymorphisms with growth traits and blood parameters was also examined. Results revealed a g.939A > G SNP (single nucleotide polymorphism) in the exon 2 exhibiting three genotypes: AA, AG, and GG. Statistical analysis indicated that this polymorphism significantly influenced blood triglyceride (P < 0.05) and cholesterol (P < 0.08) levels as well as weaning weight (P < 0.05). Animals with AG genotype had the highest blood triglyceride level and weaning weight while the highest amount of blood cholesterol was observed in animals with GG genotype. On the other hand, no significant effect was observed on birth and fat-tail weight traits. The intron 3 (g.4349A > G) was monomorphic across the studied samples. Lori-Bakhtiari breed showed significantly higher blood triglyceride and cholesterol levels, as also birth and weaning weight compared to Zel breed (P < 0.01). Considering that the literature is bereft of any report on the association study between FABP3 SNPs and sheep growth traits and blood parameters, our findings suggest that the investigated polymorphism might be one of the main genetic factors affecting growth and physiological traits in sheep.

Keywords: FABP3 gene, fatness, weaning weight, blood triglyceride, cholesterol, Zel, Lori-Bakhtiari

Procedia PDF Downloads 685

Authors: V. T. Gerov, D. I. Gerova, I. D. Micheva, N. F. Nazifova-Tasinova, M. N. Nikolova, M. G. Pasheva, B. T. Galunska

Abstract:

Multiple myeloma (MM) is the most frequent plasma cell (PC) dyscrasia that involves the skeleton. Myeloma bone disease (MBD) is characterized by osteolytic bone lesions as a result of increased osteoclasts activity not followed by reactive bone formation due to osteoblasts suppression. Skeletal complications cause significant adverse effects on quality of life and lead to increased morbidity and mortality. Last decade studies revealed the implication of different proteins in osteoclast activation and osteoblast inhibition. The aim of the present study was to determine serum levels of periostin, sRANKL and osteopontin and to evaluate their role as bone markers in MBD. Materials and methods. Thirty-two newly diagnosed MM patients (mean age: 62.2 ± 10.7 years) and 33 healthy controls (mean age: 58.9 ± 7.5 years) were enrolled in the study. According to IMWG criteria 28 patients were with symptomatic MM and 4 with monoclonal gammopathy of undetermined significance (MGUS). In respect to their bone involvement all symptomatic patients were divided into two groups (G): 9 patients with 0-3 osteolytic lesions (G1) and 19 patients with >3 osteolytic lesions and/or pathologic fractures (G2). Blood samples were drawn for routine laboratory analysis and for measurement of periostin, sRANKL and osteopontin serum levels by ELISA kits (Shanghai Sunred Biological Technology, China). Descriptive analysis, Mann-Whitney test for assessment the differences between groups and non-parametric correlation analysis were performed using GraphPad Prism v8.01. Results. The median serum levels of periostin, sRANKL and osteopontin of ММ patients were significantly higher compared to controls (554.7pg/ml (IQR=424.0-720.6) vs 396.9pg/ml (IQR=308.6-471.9), p=0.0001; 8.9pg/ml (IQR=7.1-10.5) vs 5.6pg/ml (IQR=5.1-6.4, p<0.0001 and 514.0ng/ml (IQR=469.3-754.0) vs 387.0ng/ml (IQR=335.9-441.9), p<0.0001, respectively). for assessment of differences between groups and non-parametric correlation analysis were performed using GraphPad Prism v8.01. Statistical significance was found for all tested bone markers between symptomatic MM patients and controls: G1 vs controls (p<0.03), G2 vs controls (p<0.0001) for periostin; G1 vs controls (p<0.0001), G2 vs controls (p<0.0001) for sRANKL; G1 vs controls (p=0.002), G2 vs controls (p<0.0001) for osteopontin, as well between symptomatic MM patients and MGUS patients: G1 vs MGUS (p<0.003), G2 vs MGUS (p=0.003) for periostin; G1 vs MGUS (p<0.05), G2 vs MGUS (p<0.001) for sRANKL; G1 vs MGUS (p=0.011), G2 vs MGUS (p=0.0001) for osteopontin. No differences were detected between MGUS and controls and between patients in G1 and G2 groups. Spearman correlation analysis revealed moderate positive correlation between periostin and beta-2-microglobulin (r=0.416, p=0.018), percentage bone marrow myeloma PC (r=0.432, p=0.014), and serum total protein (r=0.427, p=0.015). Osteopontin levels were also positively related to beta-2-microglobulin (r=0.540, p=0.0014), percentage bone marrow myeloma PC (r=0.423, p=0.016), and serum total protein (r=0.413, p=0.019). Serum sRANKL was only related to beta-2-microglobulin levels (r=0.398, p=0.024). Conclusion: In the present study, serum levels of periostin, sRANKL and osteopontin in newly diagnosed MM patients were evaluated. They gradually increased from MGUS to more advanced stages of MM reflecting the severity of bone destruction. These results support the idea that some new protein markers could be used in monitoring the MBD as a most severe complication of MM.

Keywords: myeloma bone disease, periostin, sRANKL, osteopontin

Procedia PDF Downloads 49

Authors: Seyhan Sahan-Firat, Meryem Temiz-Resitoglu, Demet Sinem Guden, Sefika Pinar Kucukkavruk, Bahar Tunctan, Ayse Nihal Sari, Zumrut Kocak

Abstract:

We hypothesized that mTOR inhibition may prevent the multiple organ failures following severe multiple tissue injury associated with increased NADPH oxidase system activity occur in zymosan-induced systemic inflammation. Therefore, we investigated the role of mTOR in oxidative/nitrosative stress associated with increase in NADPH oxidase activity in zymosan-induced systemic inflammation model in rats. Male Wistar rats received saline (4 ml/kg, i.p.) and zymosan (500 mg/kg, i.p.) at time 0. Saline, or zymosan-treated rats were given rapamycin (1 mg/kg, i.p.) 1 h after saline or zymosan injections. Rats were sacrified 4 h after zymosan challenge and kidney, heart, thoracic aorta, and superior mesenteric artery were collected. NADPH oxidase activity, p22phox, gp91phox, and p47phox protein expression and nitrotyrosine levels were measured in tissue samples. Zymosan administration caused an increase in NADPH oxidase activity, p22phox, gp91phox, and p47phox protein expression and nitrotyrosine levels in kidney, heart, thoracic aorta, and superior mesenteric artery. These changes caused by zymosan reversed by rapamycin, a selective mTOR inhibitor. Rapamycin alone had no effect on the parameters measured. Our results demonstrated that zymosan-induced oxidative/nitrosative stress presumably due to enhanced activity of NADPH oxidase, expression of p22phox, gp91phox, and p47phox and production of peroxynitrite were mediated by mTOR. [This work was financially supported by Research Foundation of Mersin University (2016-2-AP3-1900)].

Keywords: oxidative stress, mTOR, nitrosative stress, zymosan

Procedia PDF Downloads 309

Authors: Ka Ho Yau, Jan Frederick Engels, Kwok Kei Lai, Reinhard Renneberg

Abstract:

Lateral flow test is a prevalent technology in various sectors such as food, pharmacology and biomedical sciences. Colloidal gold (CG) is widely used as the signalling molecule because of the ease of synthesis, bimolecular conjugation and its red colour due to intrinsic SPRE. However, the production of colloidal gold is costly and requires vigorous conditions. The stability of colloidal gold are easily affected by environmental factors such as pH, high salt content etc. Silica nanoparticles are well known for its ease of production and stability over a wide range of solvents. Using reverse micro-emulsion (w/o), silica nanoparticles with different sizes can be produced precisely by controlling the amount of water. By incorporating different water-soluble dyes, a rainbow colour of the silica nanoparticles could be produced. Conjugation with biomolecules such as antibodies can be achieved after surface modification of the silica nanoparticles with organosilane. The optimum amount of the antibodies to be labelled was determined by Bradford Assay. In this work, we have demonstrated the ability of the dye-doped silica nanoparticles as a signalling molecule in lateral flow test, which showed a semi-quantitative measurement of the analyte. The image was further analysed for the LOD=10 ng of the analyte. The working range and the linear range of the test were from 0 to 2.15μg/mL and from 0 to 1.07 μg/mL (R2=0.988) respectively. The performance of the tests was comparable to those using colloidal gold with the advantages of lower cost, enhanced stability and having a wide spectrum of colours. The positives lines can be imaged by naked eye or by using a mobile phone camera for a better quantification. Further research has been carried out in multicolour detection of different biomarkers simultaneously. The preliminary results were promising as there was little cross-reactivity being observed for an optimized system. This approach provides a platform for multicolour detection for a set of biomarkers that enhances the accuracy of diseases diagnostics.

Keywords: colorimetric detection, immunosensor, paper-based biosensor, silica

Procedia PDF Downloads 375

Authors: Murali Aarthy, Sanjeev Kumar Singh

Abstract:

High risk human papillomaviruses are highly associated with the carcinoma of the cervix and the other genital tumors. Cervical cancer develops through the multistep process in which increasingly severe premalignant dysplastic lesions called cervical intraepithelial neoplastic progress to invasive cancer. The oncoprotein E7 of human papillomavirus expressed in the lower epithelial layers drives the cells into S-phase creating an environment conducive for viral genome replication and cell proliferation. The replication of the virus occurs in the terminally differentiating epithelium and requires the activation of cellular DNA replication proteins. To date, no suitable drug molecule is available to treat HPV infection whereas identification of potential drug targets and development of novel anti-HPV chemotherapies with unique mode of actions are expected. Hence, our present study aimed to identify the potential inhibitors analogous to EGCG, a green tea molecule which is considered to be safe to use for mammalian systems. A 3D similarity search on the natural small molecule library from natural product database using EGCG identified 11 potential hits based on their similarity score. The structure based docking strategies were implemented in the potential hits and the key interacting residues of protein with compounds were identified through simulation studies and binding free energy calculations. The conformational changes between the apoprotein and the complex were analyzed with the simulation and the results demonstrated that the dynamical and structural effects observed in the protein were induced by the compounds and indicated the dominance to the oncoprotein. Overall, our study provides the basis for the structural insights of the identified potential hits and EGCG and hence, the analogous compounds identified can be potent inhibitors against the HPV 16 E7 oncoprotein.

Keywords: EGCG, oncoprotein, molecular dynamics simulation, analogues

Procedia PDF Downloads 119

Authors: S. Fahiminiya, J. Nadaf, F. Rauch, L. Jerome-Majewska, J. Majewski

Abstract:

Background: Sequencing of protein coding regions of human genome (Whole Exome Sequencing; WES), has demonstrated a great success in the identification of causal mutations for several rare genetic disorders in human. Generally, most of WES studies have focused on rare variants in coding exons and splicing-sites where missense substitutions lead to the alternation of protein product. Although focusing on this category of variants has revealed the mystery behind many inherited genetic diseases in recent years, a subset of them remained still inconclusive. Here, we present the result of our WES studies where analyzing only rare variants in coding regions was not conclusive but further investigation revealed the involvement of non-coding variants and copy number variations (CNV) in etiology of the diseases. Methods: Whole exome sequencing was performed using our standard protocols at Genome Quebec Innovation Center, Montreal, Canada. All bioinformatics analyses were done using in-house WES pipeline. Results: To date, we successfully identified several disease causing mutations within gene coding regions (e.g. SCARF2: Van den Ende-Gupta syndrome and SNAP29: 22q11.2 deletion syndrome) by using WES. In addition, we showed that variants in non-coding regions and CNV have also important value and should not be ignored and/or filtered out along the way of bioinformatics analysis on WES data. For instance, in patients with osteogenesis imperfecta type V and in patients with glucocorticoid deficiency, we identified variants in 5'UTR, resulting in the production of longer or truncating non-functional proteins. Furthermore, CNVs were identified as the main cause of the diseases in patients with metaphyseal dysplasia with maxillary hypoplasia and brachydactyly and in patients with osteogenesis imperfecta type VII. Conclusions: Our study highlights the importance of considering non-coding variants and CNVs during interpretation of WES data, as they can be the only cause of disease under investigation.

Keywords: whole exome sequencing data, non-coding variants, copy number variations, rare diseases

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