Search results for: protein matrix

Authors: Sadoon Abdallah, Mizi Fan, Xiangming Zhou

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In this study, a comprehensive approach has been adopted to examine in detail the effect of various hook geometries on bond-slip characteristics. Extensive single fibre pull-out tests on ultra-high performance matrix with three different W/B ratios and embedded lengths have been carried out. Test results showed that the mechanical deformation of fibre hook is the main mechanism governing the pull-out behaviour. Furthermore, the quantitative analyses have been completed to compare the hook design contribution of 3D, 4D and 5D fibres to assess overall pull-out behaviour. It was also revealed that there is a strong relationship between the magnitude of hook contribution and W/B ratio (i.e. matrix strength). Reducing the W/B ratio from 0.20 to 0.11 greatly optimizes the interfacial transition zone (ITZ) and enables better mobilization, straightening of the hook and results in bond-slip-hardening behaviour.

Keywords: bobond mechanisms, fibre-matrix interface, hook geometry, pullout behaviour and water to binder ratio

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Authors: Jonmichael Weaver, David Miller

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Dyneema is an ultra-high molecular weight polyethylene (UHMWPE) fiber created by DSM. This fiber has many applications due to the high tensile strength, low weight, and inability to absorb water. DSM manufactures a non-woven unidirectional cross-ply [0,90]2 lamina, using the Dyneema fiber. Using this lamina system, various thickness panels are created for a 40% lighter weight alternative to Kevlar for the same ballistics protection. Environmental effects on the ply/laminate system alter the material properties, resulting in diminished ultimate performance. Understanding the specific environmental parameters and characterizing the resulting material property degradation is essential for determining the safety and reliability of Dyneema in service. Two laminas were contrasted for their response to accelerated aging by UV, humidity, and temperature cycling. Both lamina contain the same fiber, SK-99, but differ in matrix composition, Dyneema HB-210 employs a polyurethane (PUR) based matrix, and HB-212 contains a rubber-based matrix. Each system was inspected using a scanning electron microscope (SEM) and evaluated by dynamic mechanical analysis (DMA) to characterize the material property changes alongside the corresponding composite damage and matrix failure mode over the aging parameters. Overall, resulting in the HB-212 degrading faster compared with the HB-210.

Keywords: dyneema, accelerated aging, polymers, ballistics protection, armor, DSM, kevlar, composites

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Authors: Kshitij Bhardwaj, R.K. Trivedi, Shipra Dixit

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We used biological detoxification method that converts toxic residue waste of Jatropha curcas oil seeds (non edible oil seed) into industrial bio-products and animal feed material. Present study describes the complete degradation of phorbol esters by Aspergillus Niger strain during solid state fermentation (SSF) of deoiled Jatropha curcas seed cake. Phorbol esters were completely degraded in 15 days under the optimized SSF conditions viz deoiled cake 5.0 gm moistened with 5.0 ml distilled water; inoculum 2 ml of overnight grown Aspergillus niger; incubated at 30◦ C, pH 7.0. This method simultaneously induces the production of Protease enzyme by Aspergillus Niger which has high potential to be used in feedstuffs .The maximum Protease activities obtained were 709.16 mg/ml in Jatropha curcas oil seed cake. The protein isolate had small amounts of phorbol esters, phytic acid, and saponin without any lectin. Its minimum and maximum solubility were at pH 4.0&12.0. Water and oil binding capacities were 3.22 g water/g protein and 1.86 ml oil/g protein respectively.Emulsion activity showed high values in a range of basic pH. We concluded that Jatropha Curcas seed cake has a potential to be used as a novel source of functional protein for food or feed applications.

Keywords: solid state fermentation, Jatropha curcas, oil seed cake, phorbol ester

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Authors: W. P. Hu, J. V. Kumar, Jeffrey J. P. Tsai

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The inhibition of SH2 domain regulated protein-protein interactions is an attractive target for developing an effective chemotherapeutic approach in the treatment of disease. Molecular simulation is a useful tool for developing new drugs and for studying molecular recognition. In this study, we searched potential drug compounds for the inhibition of SH2 domain by performing structural similarity search in PubChem Compound Database. A total of 37 compounds were screened from the database, and then we used the LibDock docking program to evaluate the inhibition effect. The best three compounds (AP22408, CID 71463546 and CID 9917321) were chosen for MD simulations after the LibDock docking. Our results show that the compound CID 9917321 can produce a more stable protein-ligand complex compared to other two currently known inhibitors of Src SH2 domain. The compound CID 9917321 may be useful for the inhibition of SH2 domain based on these computational results. Subsequently experiments are needed to verify the effect of compound CID 9917321 on the SH2 domain in the future studies.

Keywords: nonpeptide inhibitor, Src SH2 domain, LibDock, molecular dynamics simulation

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Authors: Natalia D. Vaysfel’d, Zinaida Y. Zhuravlova

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The loaded plane elastic semistrip, the lateral boundaries of which are fixed, is considered. The integral transformations are applied directly to Lame’s equations. It leads to one dimensional boundary value problem in the transformations’ domain which is formulated as a vector one. With the help of the matrix differential calculation’s apparatus and apparatus of Green matrix function the exact solution of a vector problem is constructed. After the satisfying the boundary condition at the semi strip’s edge the problem is reduced to the solving of the integral singular equation with regard of the unknown stress at the semis trip’s edge. The equation is solved with the orthogonal polynomials method that takes into consideration the real singularities of the solution at the ends of integration interval. The normal stress at the edge of the semis trip were calculated and analyzed.

Keywords: semi strip, Green's Matrix, fourier transformation, orthogonal polynomials method

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Authors: Mateen A Khan, Taj Mohammad, Md. Imtaiyaz Hassan

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Alzheimer’s disease is characterized by the accumulation of the processing products of the amyloid beta peptide cleaved by amyloid precursor protein (APP). Iron increases the synthesis of amyloid beta peptides, which is why iron is present in Alzheimer's disease patients' amyloid plaques. Iron misregulation in the brain is linked to the overexpression of APP protein, which is directly related to amyloid-β aggregation in Alzheimer’s disease. The APP 5'-UTR region encodes a functional iron-responsive element (IRE) stem-loop that represents a potential target for modulating amyloid production. Targeted regulation of APP gene expression through the modulation of 5’-UTR sequence function represents a novel approach for the potential treatment of AD because altering APP translation can be used to improve both the protective brain iron balance and provide anti-amyloid efficacy. The molecular docking analysis of APP IRE RNA with eukaryotic translation initiation factors yields several models exhibiting substantial binding affinity. The finding revealed that the interaction involved a set of functionally active residues within the binding sites of eIF4F. Notably, APP IRE RNA and eIF4F interaction were stabilized by multiple hydrogen bonds with residues of APP IRE RNA and eIF4F. It was evident that APP IRE RNA exhibited a structural complementarity that tightly fit within binding pockets of eIF4F. The simulation studies further revealed the stability of the complexes formed between RNA and eIF4F, which is crucial for assessing the strength of these interactions and subsequent roles in the pathophysiology of Alzheimer’s disease. In addition, MD simulations would capture conformational changes in the IRE RNA and protein molecules during their interactions, illustrating the mechanism of interaction, conformational change, and unbinding events and how it may affect aggregation propensity and subsequent therapeutic implications. Our binding studies correlated well with the translation efficiency of APP mRNA. Overall, the outcome of this study suggests that the genomic modification and/or inhibiting the expression of amyloid protein by targeting APP IRE RNA can be a viable strategy to identify potential therapeutic targets for AD and subsequently be exploited for developing novel therapeutic approaches.

Keywords: Alzheimer's disease, Protein-RNA interaction analysis, molecular docking simulations, conformational dynamics, binding stability, binding kinetics, protein synthesis.

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Authors: H. Sakamoto, Y. Nakagawara, S. Oguri

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Drought stress is a critical environmental factor that adversely affects crop productivity and quality. Because of their immobile nature, plants have evolved mechanisms to sense and respond to drought stress. We identified a novel locus of Arabidopsis, designated DRA1 (drought responsive ankyrin 1), whose disruption leads to increased drought stress tolerance. DRA1 encodes a transmembrane protein with an ankyrin repeat motif that has been implicated in diverse cellular processes such as signal transduction. RT-PCR analysis revealed that there were at least two splicing variants of DRA1 transcripts in wild type plants. In response to drought stress, the levels of DRA1 transcripts retaining second and third introns were increased, whereas these introns were removed under unstressed conditions. These results suggest that DRA1 protein may negatively regulate plant drought tolerance and that the expression of DRA1 is regulated in response to drought stress by alternative splicing.

Keywords: alternative splicing, ankyrin repeat, Arabidopsis, drought tolerance

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Authors: Salman Akrokayan

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Synthetic cDNA (ScDNA) of granulocyte colony-stimulating factor (G-CSF) was constructed using a DNA synthesizer with the aim to increase its expression level. 5' end of the ScDNA of G-CSF coding region was modified by decreasing the GC content without altering the predicted amino acids sequence. The identity of the resulting protein from ScDNA was confirmed by the highly specific enzyme-linked immunosorbent assay. In conclusion, a synthetic G-CSF cDNA in combination with the recombinant DNA protocol offers a rapid and reliable strategy for synthesizing the target protein. However, the commercial utilization of this methodology requires rigorous validation and quality control.

Keywords: synthetic cDNA, recombinant G-CSF, cloning, gene expression

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Authors: M. Alsaadi, A. Erkliğ, M. Bulut

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This paper experimentally investigates the flexural and tensile properties of the industrial wastes sewage sludge ash (SSA) and fly ash (FA), and conventional ceramic powder silicon carbide (SiC) filled polyester composites. Four weight fractions (5, 10, 15 and 20 wt%) for each micro filler were used for production of composites. Then, test samples were produced according to ASTM. The resulting degree of particle dispersion in the polymer matrix was visualized by using scanning electron microscope (SEM). Results from this study showed that the tensile strength increased up to its maximum value at filler content 5 wt% of SSA, FA and SiC. Flexural strength increased with addition of particulate filler up to its maximum value at filler content 5 wt% of SSA and FA while for SiC decreased for all weight fractions gradually. The addition of SSA, FA and SiC fillers resulted in increase of tensile and flexural modulus for all the particulate composites. Industrial waste SSA can be used as an additive with polymer to produce composite materials.

Keywords: particle-reinforcement, sewage sludge ash, polymer matrix composites, mechanical properties

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Authors: Pawan Kumar Sharma, J. Stephan Sampath Kumar, S. Anand, Chandana B. L.

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Protein is the major component for the success in culture of shrimp and fishes. Apparently, excess dietary protein is undesirable, as it not only enhances the production cost but also leads to water quality deterioration. A field survey was conducted with aqua farmers of Kerala, India, a leading state in coastal aquaculture, to assess the role of protein component in feed that can be efficiently and effectively managed for sustainable aquaculture. The study showed an average feed amount of 13.55 ± 2.16 tonnes per hectare was being used by the farmers of Kerala. The average feed cost percentage of Rs. 57.76 ± 13.46 /kg was invested for an average protein level of 36.26 % ± 0.082 in the feed and Rs.78.95 ± 3.086 per kilogram of feed was being paid by the farmers. Study revealed that replacement of fish meal and fish oil within shrimp aquafeeds with alternative protein, and lipid sources can only be achieved if changes are made in the basic shrimp culturing practices, such as closed farming system through water recycling or zero-water exchange, and by maximizing in-situ, floc and natural food production within the culture system. The upshot of such production systems is that imports of high-quality feed ingredients and aqua feeds can eventually be eliminated, and the utilization of locally available feed ingredients from agricultural by-products can be greatly improved and maximized. The promotion of closed shrimp production systems would also greatly reduce water use and increase shrimp production per unit area but would necessitate the continuous provision of electricity for aeration during production. Alternative energy sources such as solar power might be used, and resource poor farming communities should also explore wind energy for use. The study concluded that farm made feed and closed farming systems are essential for the sustainability and profitability of the aquaculture industry.

Keywords: aqua feeds, floc, fish meal, protein, zero-water exchange

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Authors: Mohammad M. Khan, Gajendra Dixit

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Partially lubricated sliding wear behaviour of a zinc-based alloy reinforced with 10wt% SiC particles has been studied as a function of applied load and solid lubricant particle size and has been compared with that of matrix alloy and conventionally used grey cast iron. The wear tests were conducted at the sliding velocities of 2.1m/sec in various partial lubricated conditions using pin on disc machine as per ASTM G-99-05. Base oil (SAE 20W-40) or mixture of the base oil with 5wt% graphite of particle sizes (7-10 µm) and (100 µm) were used for creating lubricated conditions. The matrix alloy revealed primary dendrites of a and eutectoid a + h and Î phases in the Inter dendritic regions. Similar microstructure has been depicted by the composite with an additional presence of the dispersoid SiC particles. In the case of cast iron, flakes of graphite were observed in the matrix; the latter comprised of (majority of) pearlite and (limited quantity of) ferrite. Results show a large improvement in wear resistance of the zinc-based alloy after reinforcement with SiC particles. The cast iron shows intermediate response between the matrix alloy and composite. The solid lubrication improved the wear resistance and friction behaviour of both the reinforced and base alloy. Moreover, minimum wear rate is obtained in oil+ 5wt % graphite (7-10 µm) lubricated environment for the matrix alloy and composite while for cast iron addition of solid lubricant increases the wear rate and minimum wear rate is obtained in case of oil lubricated environment. The cast iron experienced higher frictional heating than the matrix alloy and composite in all the cases especially at higher load condition. As far as friction coefficient is concerned, a mixed trend of behaviour was noted. The wear rate and frictional heating increased with load while friction coefficient was affected in an opposite manner. Test duration influenced the frictional heating and friction coefficient of the samples in a mixed manner.

Keywords: solid lubricant, sliding wear, grey cast iron, zinc based metal matrix composites

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Authors: Alan C. Lin, Tzu-Kuan Lin, Tsong Der Lin

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This paper proposes a new method to find the equations of transformation matrix for the rotation angles of the two rotational axes and the coordinates of the three linear axes of an orthogonal multi-axis milling machine. This approach provides intuitive physical meanings for rotation angles of multi-axis machines, which can be used to evaluate the accuracy of the conversion from CL data to NC data.

Keywords: CAM, multi-axis milling machining, transformation matrix, rotation angles

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Authors: Boyu Li

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Mixed matrix membranes (MMMs), as a separation technology, can improve CO₂ recycling efficiency and reduce the environmental impacts associated with huge emissions. Nevertheless, many challenges must be overcome to design excellent selectivity and permeability performance MMMs. Herein, this work demonstrates the design of nano-scale GNPs (Cu-BDC@PEG) with strong compatibility and high free friction volume (FFV) is an effective way to construct non-interfacial voids MMMs with a desirable combination of selectivity and permeability. Notably, the FFV boosted thanks to the chain length and shape of the GNPs. With this, the permeability and selectivity of Cu-BDC@PEG/PVDF MMMs had also been significantly improved. As such, compatible Cu-BDC@PEG proves very efficient for resolving challenges of MMMs with poor compatibility on the basis of the interfacial defect. Poly (Ethylene Glycol) (PEG) with oxygen groups can be finely coordinated with Cu-MOFs to disperse Cu-BDC@PEG homogenously and form hydrogen bonds with matrix to achieve continuous phase. The resultant MMMs exhibited a simultaneous enhancement of gas permeability (853.1 Barrer) and ideal CO₂/N selectivity (41.7), which has surpassed Robenson's upper bound. Moreover, Cu-BDC@PEG/PVDF has a high-temperature resistance and a long time sustainably. This attractive separation performance of Cu-BDC@PEG/PVDF offered an exciting platform for the development of composite membranes for sustainable CO₂ separations.

Keywords: metal organic framework, CO₂ separation, mixed matrix membrane, polymer

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Authors: Harendra Singh, Rajesh Pandey

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The paper presents a numerical method based on operational matrix of integration and Ryleigh method for the solution of a class of non-linear fractional variational problems (NLFVPs). Chebyshev first kind polynomials are used for the construction of operational matrix. Using operational matrix and Ryleigh method the NLFVP is converted into a system of non-linear algebraic equations, and solving these equations we obtained approximate solution for NLFVPs. Convergence analysis of the proposed method is provided. Numerical experiment is done to show the applicability of the proposed numerical method. The obtained numerical results are compared with exact solution and solution obtained from Chebyshev third kind. Further the results are shown graphically for different fractional order involved in the problems.

Keywords: non-linear fractional variational problems, Rayleigh-Ritz method, convergence analysis, error analysis

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Authors: Natalia Caldeira De Carvalho, Tassia Batista Pessato, Luis Gustavo R. Fernandes, Ricardo L. Zollner, Flavia Maria Netto

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Protein hydrolysates are ingredients of enteral diets and hypoallergenic formulas. Enzymatic hydrolysis is the most commonly used method for reducing the antigenicity of milk protein. The antigenicity and physicochemical characteristics of the protein hydrolysates depend on the reaction parameters. Among them, pH has been pointed out as of the major importance. Hydrolysis reaction in laboratory scale is commonly carried out under controlled pH (pH-stat). However, from the industrial point of view, controlling pH during hydrolysis reaction may be infeasible. This study evaluated the impact of pH control on the physicochemical properties and antigenicity of the hydrolysates of whey proteins with Alcalase. Whey protein isolate (WPI) solutions containing 3 and 7 % protein (w/v) were hydrolyzed with Alcalase 50 and 100 U g-1 protein at 60°C for 180 min. The reactions were carried out under controlled and uncontrolled pH conditions. Hydrolyses performed under controlled pH (pH-stat) were initially adjusted and maintained at pH 8.5. Hydrolyses carried out without pH control were initially adjusted to pH 8.5. Degree of hydrolysis (DH) was determined by OPA method, peptides profile was evaluated by HPLC-RP, and molecular mass distribution by SDS-PAGE/Tricine. The residual α-lactalbumin (α-La) and β-lactoglobulin (β-Lg) concentrations were determined using commercial ELISA kits. The specific IgE and IgG binding capacity of hydrolysates was evaluated by ELISA technique, using polyclonal antibodies obtained by immunization of female BALB/c mice with α-La, β-Lg and BSA. In hydrolysis under uncontrolled pH, the pH dropped from 8.5 to 7.0 during the first 15 min, remaining constant throughout the process. No significant difference was observed between the DH of the hydrolysates obtained under controlled and uncontrolled pH conditions. Although all hydrolysates showed hydrophilic character and low molecular mass peptides, hydrolysates obtained with and without pH control exhibited different chromatographic profiles. Hydrolysis under uncontrolled pH released, predominantly, peptides between 3.5 and 6.5 kDa, while hydrolysis under controlled pH released peptides smaller than 3.5 kDa. Hydrolysis with Alcalase under all conditions studied decreased by 99.9% the α-La and β-Lg concentrations in the hydrolysates detected by commercial kits. In general, β-Lg concentrations detected in the hydrolysates obtained under uncontrolled pH were significantly higher (p<0.05) than those detected in hydrolysates produced with pH control. The anti-α-La and anti-β-Lg IgE and IgG responses to all hydrolysates decreased significantly compared to WPI. Levels of specific IgE and IgG to the hydrolysates were below 25 and 12 ng ml-1, respectively. Despite the differences in peptide composition and α-La and β-Lg concentrations, no significant difference was found between IgE and IgG binding capacity of hydrolysates obtained with or without pH control. These results highlight the impact of pH on the hydrolysates characteristics and their concentrations of antigenic protein. Divergence between the antigen detection by commercial ELISA kits and specific IgE and IgG binding response was found in this study. This result shows that lower protein detection does not imply in lower protein antigenicity. Thus, the use of commercial kits for allergen contamination analysis should be cautious.

Keywords: allergy, enzymatic hydrolysis, milk protein, pH conditions, physicochemical characteristics

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Authors: I. Sani, A. Abdulhamid, F. Bello, Isah M. Fakai

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This research has focused on the application of green fluorescent protein (GFP) as a new technique for direct monitoring of fermentation processes involving cultured bacteria. To use GFP as a sensor for pH and oxygen, percentage ratio of red fluorescence to green (% R/G) was evaluated. Assessing the magnitude of the % R/G ratio in relation to low or high pH and oxygen concentration, the bacterial strains were cultivated under aerobic and anaerobic conditions. SCC1 strains of E. coli were grown in a 5 L laboratory fermenter, and during the fermentation, the pH and temperature were controlled at 7.0 and 370C respectively. Dissolved oxygen tension (DOT) was controlled between 15-100% by changing the agitation speed between 20-500 rpm respectively. Effect of reducing the DOT level from 100% to 15% was observed after 4.5 h fermentation. There was a growth arrest as indicated by the decrease in the OD650 at this time (4.5-5 h). The relative fluorescence (green) intensity was decreased from about 460 to 420 RFU. However, %R/G ratio was significantly increased from about 0.1% to about 0.25% when the DOT level was decreased to 15%. But when the DOT was changed to 100%, a little increase in the RF and decrease in the %R/G ratio were observed. Therefore, GFP can effectively detect and indicate any change in pH and oxygen level during fermentation processes.

Keywords: Escherichia coli SCC1, fermentation process, green fluorescent protein, red fluorescence

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Authors: Mohammed Alasel, Michael Keusgen

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Gold is a noble metal; in its nano-scale level (e.g. spherical nanoparticles), the conduction electrons are triggered to collectively oscillate with a resonant frequency when certain wavelengths of electromagnetic radiation interact with its surface; this phenomenon is known as surface plasmon resonance (SPR). SPR is responsible for giving the gold nanoparticles its intense red color depending mainly on its size, shape and distance between nanoparticles. A decreased distance between gold nanoparticles results in aggregation of them causing a change in color from red to blue. This aggregation enables gold nanoparticles to serve as a sensitive biosensoric indicator. In the proposed work, gold nanoparticles were modified with two proteins: i) Borrelia antigen, variable lipoprotein surface-exposed protein (VlsE), and ii) protein A. VlsE antigen induces a strong antibody response against Lyme disease and can be detected from early to late phase during the disease in humans infected with Borrelia. In addition, it shows low cross-reaction with the other non-pathogenic Borrelia strains. The high specificity of VlsE antigen to anti-Borrelia antibodies, combined simultaneously with the high specificity of protein A to the Fc region of all IgG human antibodies, was utilized to develop a rapid test for serological point of care diagnosis of borreliosis in human serum. Only in the presence of anti-Borrelia antibodies in the serum probe, an aggregation of gold nanoparticles can be observed, which is visible by a concentration-dependent colour shift from red (low IgG) to blue (high IgG). Experiments showed it is clearly possible to distinguish between positive and negative sera samples using a simple suspension of the two-protein modified gold nanoparticles in a very short time (30 minutes). The proposed work showed the potential of using such modified gold nanoparticles generally for serological diagnosis. Improved specificity and reduced assay time can be archived in applying increased salt concentrations combined with decreased pH values (pH 5).

Keywords: gold nanoparticles, gold aggregation, serological diagnosis, protein A, lyme borreliosis

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Authors: Kübra Aktaş, Nihat Akin

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Tarhana is a traditional Turkish fermented food. It is widely consumed as soup and includes many different ingredients such as wheat flour, various vegetables, and spices, yoghurt, bakery yeast. It can also be enriched by adding other ingredients. Thus, its nutritional properties can be enhanced. In this study, tarhana was supplemented with two different types of brans (rice bran and corn bran) and WPC (whey protein concentrate powder) to improve its nutritional and functional properties. Some chemical properties of tarhana containing two different brans and their levels (0, 5, 10 and 15%) and WPC (0, 5, 10%) were investigated. The results indicated that addition of WPC increased ash content in tarhanas which were fortified with rice and corn bran. The highest antioxidant and phenolic content values were obtained with addition of rice bran in tarhana formulation. Compared to tarhana with corn bran, rice bran addition gave higher oil content values. The cellulose content of tarhana samples was determined between 0.75% and 2.74% and corn bran showed an improving effect on cellulose contents of samples. In terms of protein content, addition of WPC into the tarhana raised protein content for the samples.

Keywords: corn, rice, tarhana, whey

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Authors: Katerina H. Takova, Ivan N. Minkov, Gergana G. Zahmanova

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Molecular farming is the production of recombinant proteins in plants with the aim to use the protein as a purified product, crude extract or directly in the planta. Plants gain more attention as expression systems compared to other ones due to the cost effective production of pharmaceutically important proteins, appropriate post-translational modifications, assembly of complex proteins, absence of human pathogens to name a few. In addition, transient expression in plant leaves enables production of recombinant proteins within few weeks. Hepatitis E virus (HEV) is a causative agent of acute hepatitis. HEV causes epidemics in developing countries and is primarily transmitted through the fecal-oral route. Presently, all efforts for development of Hepatitis E vaccine are focused on the Open Read Frame 2 (ORF2) capsid protein as it contains epitopes that can induce neutralizing antibodies. For our purpose, we used the CMPV-based vector-pEAQ-HT for transient expression of HEV ORF2 in Nicotiana benthamina. Different molecular analysis (Western blot and ELISA) showed that HEV ORF2 capsid protein was expressed in plant tissue in high-yield up to 1g/kg of fresh leaf tissue. Electron microscopy showed that the capsid protein spontaneously assembled in low abundance virus-like particles (VLPs), which are highly immunogenic structures and suitable for vaccine development. The expressed protein was recognized by both human and swine HEV positive sera and can be used as a diagnostic reagent for the detection of HEV infection. Production of HEV capsid protein in plants is a promising technology for further HEV vaccine investigations. Here, we reported for a rapid high-yield transient expression of a recombinant protein in plants suitable for vaccine production as well as a diagnostic reagent. Acknowledgments -The authors’ research on HEV is supported with grants from the Project PlantaSYST under the Widening Program, H2020 as well as under the UK Biotechnological and Biological Sciences Research Council (BBSRC) Institute Strategic Programme Grant ‘Understanding and Exploiting Plant and Microbial Secondary Metabolism’ (BB/J004596/1). The authors want to thank Prof. George Lomonossoff (JIC, Norwich, UK) for his contribution.

Keywords: hepatitis E virus, plant molecular farming, transient expression, vaccines

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Authors: Muhammad Arifur Rahman, Talukdar M. Waliullah, Takashi Ushimaru

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Nucleophagy selectively degrades nuclear materials, especially nucleolus after nutrient starvation or inactivation of TORC1 kinase in budding yeast. Budding yeast phosphatidate (PA) phosphatase Pah1 that converts PA to diacylglycerol is essential for partitioning of lipid precursors between membrane and storage that is crucial for many aspects of cell growth and development. Pah1 is required for nuclear/ER membrane biogenesis and vacuole function, but whether Pah1 and its activator Nem1/Spo7 protein phosphatase complex are involved in autophagy is largely unknown. Loss of Pah1 causes expansion of the nucleus and fragmentation of the vacuole. Here we show that Pah1 is required for bulk autophagy and nucleophagy after TORC1 inactivation. Loss of Pah1 impaired nucleophagy severely and bulk autophagy to a lesser extent. Loss of the Pah1 activator Nem1-Spo7 protein phosphatase exhibited similar features.

Keywords: autophagy, Nem1/Spo7 phosphatase, Pah1, nucleophagy, TORC1

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Authors: Nijole Savickiene, Antonella Di Sotto, Gabriela Mazzanti, Rasa Starselskyte, Silvia Di Giacomo, Annabella Vitalone

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Plant lectins are non-enzymic and non-immune origin proteins that specifically recognize and bind to various sugar structures and possess the activity to agglutinate cells and/or precipitate polysaccharides and glycoconjugates. The emerging evidences showed that plant lectins contribute not only to tumour cell recognition but also to cell adhesion and localization, to signal transduction, to mitogenic cytotoxicity and apoptosis. Among chitin-binding lectins, the Urtica dioica agglutinin (UDA), which is a complex of different isoforms, has been poorly studied for its biological activity. In this context and according to the increasing interest for lectins as novel antitumor drugs, present paper aimed at evaluating the potential antimutagenic activity of a lectin-like glycoprotein-enriched fraction from aerial part of Urtica dioica L. Aim: to evaluate the potential chemopreventive properties of a protein - lectin fraction from the aerial part of Urtica dioica. Materials and methods: Protein – lectin fraction has been tested for the antimutagenic activity in bacteria (50–800 mg/plate; Ames test by the preincubation method) and for the cytotoxicity on human hepatoma HepG2 cells (0.06–2 mg/mL; 24 and 48 h incubation). Results: Protein – lectin fraction from stinging nettle was not cytotoxic on HepG2 cells up to 2 mg/mL; conversely, it exhibited a strong antimutagenic activity against the mutagen 2-aminoanthracene (2AA) in all strains tested (maximum inhibition of 56.78 and 61% in TA98, TA100, and WP2uvrA strains, respectively, at 800 mg/plate). Discussion and conclusions: Protein – lectin fraction from Urtica dioica L. possesses antimutagenic and radical scavenging properties. Being 2AA a pro-carcinogenic agent, we hypothesize that the antimutagenicity of it can be due to the inhibition of CYP450-isoenzymes, involved in the mutagen bioactivation.

Keywords: lectins, antimutagenicity, chemoprevention, Urtica dioica

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Authors: Taweechai Nuntawisuttiwong, Natasha Dejdumrong

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Geometric modeling plays an important role in the constructions and manufacturing of curve, surface and solid modeling. Their algorithms are critically important not only in the automobile, ship and aircraft manufacturing business, but are also absolutely necessary in a wide variety of modern applications, e.g., robotics, optimization, computer vision, data analytics and visualization. The calculation and display of geometric objects can be accomplished by these six techniques: Polynomial basis, Recursive, Iterative, Coefficient matrix, Polar form approach and Pyramidal algorithms. In this research, the coefficient matrix (simply called monomial form approach) will be used to model polynomial rectangular patches, i.e., Said-Ball, Wang-Ball, DP, Dejdumrong and NB1 surfaces. Some examples of the monomial forms for these surface modeling are illustrated in many aspects, e.g., construction, derivatives, model transformation, degree elevation and degress reduction.

Keywords: monomial forms, rectangular surfaces, CAGD curves, monomial matrix applications

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Authors: Nurul Na'imy Wan, Mohamad Sazali Said, Jaharah Ab. Ghani, Rusli Othman

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This paper presents the surface roughness of the aluminium silicon alloy (AlSi) matrix composite which has been reinforced with aluminium nitride (AlN). Experiments were conducted at various cutting speeds, feed rates, and depths of cut, according to a standard orthogonal array L27 of Taguchi method using TiN coating tool of insert. The signal-to-noise (S/N) ratio and analysis of variance are applied to study the characteristic performance of cutting speeds, feed rates and depths of cut in measuring the surface roughness during the milling operation. The surface roughness was observed using Mitutoyo Formtracer CS-500 and analyzed using the Taguchi method. From the Taguchi analysis, it was found that cutting speed of 230 m/min, feed rate of 0.4 mm/tooth, depth of cut of 0.3 mm were the optimum machining parameters using TiN coating insert.

Keywords: AlSi/AlN metal matrix composite (MMC), surface roughness, Taguchi method, machining parameters

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Authors: Tzan-Chain Lee

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Seeds are non-leaves green tissues. Photosynthesis begins with light absorption by chlorophyll and then the energy transfer between two pigment-protein complexes (PPC). Most studies of photosynthesis and PPC expression were focused on leaves; however, during seeds’ development were rare. Developed seeds from beginning pod (stage R3) to dried seed (stage R8), and the dried seed after sowing for 1-4 day, were analyzed for their chlorophyll contents. Thornber and MARS gel systems analysis compositions of PPC. Chlorophyll fluorescence was used to detect maximal photosynthetic efficiency (Fv/Fm). During soybean and black soybean seeds development (stages R3-R6), Fv/Fm up to 0.8, and then down-regulated after full seed (stage R7). In dried seed (stage R8), the two plant seeds lost photosynthetic activity (Fv/Fm=0), but chlorophyll degradation only occurred in soybean after full seed. After seeds sowing for 4 days, chlorophyll drastically increased in soybean seeds, and Fv/Fm recovered to 0.8 in the two seeds. In PPC, the two soybean seeds contained all PPC during seeds development (stages R3-R6), including CPI, CPII, A1, AB1, AB2, and AB3. However, many proteins A1, AB1, AB2, and CPI were totally missing in the two dried seeds (stage R8). The deficiency of these proteins in dried seeds might be caused by the incomplete photosynthetic activity. After seeds germination and seedling exposed to light for 4 days, all PPC were recovered, suggesting that completed PPC took place in the two soybean seeds. This study showed the reversibility of photosynthetic activity and pigment-protein complexes during soybean and black soybean seeds development.

Keywords: light-harvesting complex, pigment–protein complexes, soybean cotyledon, grana development

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Authors: A. G. Pershina, P. S. Postnikov, M. E. Trusova, D. O. Burlakova, A. E. Sazonov

Abstract:

Invention of magnetic-fluorescent nanocomposites is a rapidly developing area of research. The magnetic-fluorescent nanocomposite attractiveness is connected with the ability of simultaneous management and control of such nanocomposites by two independent methods based on different physical principles. These nanocomposites are applied for the solution of various essential scientific and experimental biomedical problems. The aim of this research is development of principle approach to nanobiohybrid structures with magnetic and fluorescent properties design. The surface of carbon-coated iron nanoparticles (Fe@C) were covalently modified by 4-carboxy benzenediazonium tosylate. Recombinant fluorescent protein TagGFP2 (Eurogen) was obtained in E. coli (Rosetta DE3) by standard laboratory techniques. Immobilization of TagGFP2 on the nanoparticles surface was provided by the carbodiimide activation. The amount of COOH-groups on the nanoparticle surface was estimated by elemental analysis (Elementar Vario Macro) and TGA-analysis (SDT Q600, TA Instruments. Obtained nanocomposites were analyzed by FTIR spectroscopy (Nicolet Thermo 5700) and fluorescence microscopy (AxioImager M1, Carl Zeiss). Amount of the protein immobilized on the modified nanoparticle surface was determined by fluorimetry (Cary Eclipse) and spectrophotometry (Unico 2800) with the help of preliminary obtained calibration plots. In the FTIR spectra of modified nanoparticles the adsorption band of –COOH group around 1700 cm-1 and bands in the region of 450-850 cm-1 caused by bending vibrations of benzene ring were observed. The calculated quantity of active groups on the surface was equal to 0,1 mmol/g of material. The carbodiimide activation of COOH-groups on nanoparticles surface results to covalent immobilization of TagGFP2 fluorescent protein (0.2 nmol/mg). The success of immobilization was proved by FTIR spectroscopy. Protein characteristic adsorption bands in the region of 1500-1600 cm-1 (amide I) were presented in the FTIR spectrum of nanocomposite. The fluorescence microscopy analysis shows that Fe@C-TagGFP2 nanocomposite possesses fluorescence properties. This fact confirms that TagGFP2 protein retains its conformation due to immobilization on nanoparticles surface. Magnetic-fluorescent nanocomposite was obtained as a result of unique design solution implementation – the fluorescent protein molecules were fixed to the surface of superparamagnetic carbon-coated iron nanoparticles using original diazonium salts.

Keywords: carbon-coated iron nanoparticles, diazonium salts, fluorescent protein, immobilization

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Authors: Manel Hannachi, Mustapha Nechiche, Said Azem

Abstract:

This study focuses on biodegradable materials made from Poly-lactic acid (PLA) and vegetal reinforcements. Three materials are developed from PLA, as a matrix, and : (i) olive kernels (OK); (ii) alfa (α) short fibers and (iii) OK+ α mixture, as reinforcements. After processing of PLA pellets and olive kernels in powder and alfa stems in short fibers, three mixtures, namely PLA-OK, PLA-α, and PLA-OK-α are prepared and homogenized in Turbula®. These mixtures are then compacted at 180°C under 10 MPa during 15 mn. Scanning Electron Microscopy (SEM) examinations show that PLA matrix adheres at surface of all reinforcements and the dispersion of these ones in matrix is good. X-ray diffraction (XRD) analyses highlight an increase of PLA inter-reticular distances, especially for the PLA-OK case. These results are explained by the dissociation of some molecules derived from reinforcements followed by diffusion of the released atoms in the structure of PLA. This is consistent with Fourier Transform Infrared Spectroscopy (FTIR) and Differential Scanning Calorimetry (DSC) analysis results.

Keywords: alfa short fibers, biodegradable composite, olive kernels, poly-lactic acid

Procedia PDF Downloads 149

Authors: Othman Elmahdy Othman

Abstract:

The objectives of this study were to identify polymorphisms in the STAT5A using Restriction Fragment Length Polymorphism and DGAT1 using Single-Strand Conformation Polymorphism genes among three Egyptian goat breeds (Barki, Zaraibi, and Damascus) as well as investigate the effect of their genotypes on milk composition traits of Zaraibi goats. One hundred and fifty blood samples were collected for DNA extraction, 60 from Zaraibi, 40 from Damascus and 50 from Barki breeds. Fat, protein and lactose percentages were determined in Zaraibi goat milk using an automatic milk analyzer. Two genotypes, CC and CT (for STAT5A) and C-C- and C-C+ (for DGAT1), were identified in the three Egyptian goat breeds with different frequencies. The associations between these genotypes and milk fat, protein and lactose were determined in Zaraibi breed. The results showed that the STAT5A genotypes had significant effects on milk yield, protein, fat and lactose with the superiority of CT genotype over CC. Regarding DGAT1 polymorphism, the result showed the only association between it with milk fat where the animals with C-C+ genotype had greater milk fat than animals possess C-C- genotype. The association of combined genotypes with milk trait declared that the does with heterozygous genotypes for both genes are preferred than does with homozygous genotypes where the animals with CTC-C+ have more milk yield, fat and protein than those with CCC-C- genotype. In conclusion, the result showed that C/T and C-/C+ SNPs of STAT5A and DGAT1 genes respectively may be useful markers for assisted selection programs to improve goat milk composition

Keywords: DGAT1, genetic polymorphism, milk trait, STAT5A

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Authors: Mohammad M. Khan, Pankaj Agarwal

Abstract:

The present investigation deals with the microstructures, mechanical and detailed wear characteristics of in situ TiC particles reinforced zinc aluminum-based metal matrix composites. The composites have been synthesized by liquid metallurgy route using vortex technique. The composite was found to be harder than the matrix alloy due to high hardness of the dispersoid particles therein. The former was also lower in ultimate tensile strength and ductility as compared to the matrix alloy. This could be explained to be due to the use of coarser size dispersoid and larger interparticle spacing. Reasonably uniform distribution of the dispersoid phase in the alloy matrix and good interfacial bonding between the dispersoid and matrix was observed. The composite exhibited predominantly brittle mode of fracture with microcracking in the dispersoid phase indicating effective easy transfer of load from matrix to the dispersoid particles. To study the wear behavior of the samples three different types of tests were performed namely: (i) sliding wear tests using a pin on disc machine under dry condition, (ii) high stress (two-body) abrasive wear tests using different combinations of abrasive media and specimen surfaces under the conditions of varying abrasive size, traversal distance and load, and (iii) low-stress (three-body) abrasion tests using a rubber wheel abrasion tester at various loads and traversal distances using different abrasive media. In sliding wear test, significantly lower wear rates were observed in the case of base alloy over that of the composites. This has been attributed to the poor room temperature strength as a result of increased microcracking tendency of the composite over the matrix alloy. Wear surfaces of the composite revealed the presence of fragmented dispersoid particles and microcracking whereas the wear surface of matrix alloy was observed to be smooth with shallow grooves. During high-stress abrasion, the presence of the reinforcement offered increased resistance to the destructive action of the abrasive particles. Microcracking tendency was also enhanced because of the reinforcement in the matrix. The negative effect of the microcracking tendency was predominant by the abrasion resistance of the dispersoid. As a result, the composite attained improved wear resistance than the matrix alloy. The wear rate increased with load and abrasive size due to a larger depth of cut made by the abrasive medium. The wear surfaces revealed fine grooves, and damaged reinforcement particles while subsurface regions revealed limited plastic deformation and microcracking and fracturing of the dispersoid phase. During low-stress abrasion, the composite experienced significantly less wear rate than the matrix alloy irrespective of the test conditions. This could be explained to be due to wear resistance offered by the hard dispersoid phase thereby protecting the softer matrix against the destructive action of the abrasive medium. Abraded surfaces of the composite showed protrusion of dispersoid phase. The subsurface regions of the composites exhibited decohesion of the dispersoid phase along with its microcracking and limited plastic deformation in the vicinity of the abraded surfaces.

Keywords: abrasive wear, liquid metallurgy, metal martix composite, SEM

Procedia PDF Downloads 154

Authors: Hue Dinh, Binh Nguyen, Vivian Mendez, Phillip W. Taylor, Fleur Ponton

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Better understanding of how parental diet affects offspring traits is an important ecological and evolutionary question. In this study, we explored how maternal diet influences offspring physiology and resistance to infection using Bactrocera tryoni (Q-fly) as a system model. Female Q-flies were fed one of six single diets varying in their yeast-to-sugar ratio yielding six protein-to-carbohydrate ratios. As controls, we used females that were given a choice between yeast and sugar. Males were reared on a choice diet and allowed to mate with females 14 days post-emergence. Results showed that while maternal diet does not influence offspring developmental time, it has a strong effect on larval body weight. Mother fed either high-protein or high-sugar diet produced larger progeny. By challenging offspring with the bacterium Serratia marcescens, we found that female offspring from mothers fed high-sugar diet survived better the infection compared to those from mothers fed low-sugar diet. In contrast, male offspring produced by mother fed high-protein diet showed better resistance to the infection compared to those produced by mother fed low-protein diet. These results suggested sex-dependent transgenerational effects of maternal nutrition on offspring physiology and immunity.

Keywords: bacterial infection, Bactrocera tryoni, maternal diet, offspring, Serretia marcescens

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Authors: Tarana Siddika, Ilka U. Heinemann, Patrick O’Donoghue

Abstract:

Protein kinase B (AKT1) is a serine/threonine kinase and central transducer of cell survival pathways. Typical approaches to study AKT1 biology in cells rely on growth factor or insulin stimulation that activates AKT1 via phosphorylation at two key regulatory sites (Threonine308, Serine473), yet cell stimulation also activates many other kinases and fails to differentiate the effect of the two main activating sites of AKT1 on downstream substrate phosphorylation and cell growth. While both AKT1 activating sites are associated with disease and used as clinical markers, in some cancers, high levels of Threonine308 phosphorylation are associated with poor prognosis while in others poor survival correlates with high Serine473 levels. To produce cells with specific AKT1 activity, a system was developed to deliver active AKT1 to human cells. AKT1 phospho-variants were produced from Escherichia coli with programmed phosphorylation by genetic code expansion. Tagging of AKT1 with an N-terminal cell penetrating peptide tag derived from the human immunodeficiency virus trans-activator of transcription (TAT) helped to enter AKT1 proteins in mammalian cells. The TAT-tag did not alter AKT1 kinase activity and was necessary and sufficient to rapidly deliver AKT1 protein variants that persisted in human cells for 24 h without the need to use transfection reagents. TAT-pAKT1T308, TAT-pAKT1S473 and TAT-pAKT1T308S473 proteins induced selective phosphorylation of the known AKT1 substrate GSK-3αβ, and downstream stimulation of the AKT1 pathway as evidenced by phosphorylation of ribosomal protein S6 at Serine240/244 in transfected cells. Increase in cell growth and proliferation was observed due to the transfection of different phosphorylated AKT1 protein variants compared to cells with TAT-AKT1 protein. The data demonstrate efficient delivery of AKT1 with programmed phosphorylation to human cells, thus establishing a cell-based model system to investigate signaling that is dependent on specific AKT1 activity and phosphorylation.

Keywords: cell penetrating peptide, cell signaling, protein kinase b (AKT1), phosphorylation

Procedia PDF Downloads 122