Search results for: core protein

Authors: Lavinia Ruta, Ileana Farcasanu

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The yeast surface display (YSD) technique enables the expression of proteins on yeast cell surfaces, facilitating the identification and isolation of proteins with targeted binding properties, such as nanobodies. Nanobodies, derived from camelid species, are single-domain antibody fragments renowned for their high affinity and specificity towards target proteins, making them valuable in research and potentially in therapeutics. Their advantages include a compact size (~15 kDa), robust stability, and the ability to target challenging epitopes. The project endeavors to establish and validate a platform for producing Green Fluorescent Protein (GFP) and mCherry nanobodies using the yeast surface display method. mCherry, a prevalent red fluorescent protein sourced from coral species, is commonly utilized as a genetic marker in biological studies due to its vibrant red fluorescence. The GFP-nanobody, a single variable domain of heavy-chain antibodies (VHH), exhibits specific binding to GFP, offering a potent means for isolating and engineering fluorescent protein fusions across various biological research domains. Both GFP and mCherry nanobodies find specific utility in cellular imaging and protein analysis applications.

Keywords: YSD, nanobodies, GFP, Saccharomyces cerevisiae

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Authors: Vineeta Kaushik, Manisha Goel

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Chaperones are proteins that help other proteins fold correctly, and are found in all domains of life i.e., prokaryotes, eukaryotes and archaea. Various comparative genomic studies have suggested that the archaeal protein folding machinery appears to be highly similar to that found in eukaryotes. In case of protein folding; slow rotation of peptide prolyl-imide bond is often the rate limiting step. Formation of the prolyl-imide bond during the folding of a protein requires the assistance of other proteins, termed as peptide prolyl cis-trans isomerases (PPIases). Cyclophilins constitute the class of peptide prolyl isomerases with a wide range of biological function like protein folding, signaling and chaperoning. Most of the cyclophilins exhibit PPIase enzymatic activity and play active role in substrate protein folding which classifies them as a category of molecular chaperones. Till date, there is not very much data available in the literature on archaeal cyclophilins. We aim to compare the structural and biochemical features of the cyclophilin protein from within the three domains to elucidate the features affecting their stability and enzyme activity. In the present study, we carry out in-silico analysis of the cyclophilin proteins to predict their conserved residues, sites under positive selection and compare these proteins to their bacterial and eukaryotic counterparts to predict functional divergence. We also aim to clone and express these proteins in heterologous system and study their biophysical characteristics in detail using techniques like CD and fluorescence spectroscopy. Overall we aim to understand the features contributing to the folding, stability and dynamics of the archaeal cyclophilin proteins.

Keywords: biophysical characterization, x-ray crystallography, chaperone-like activity, cyclophilin, PPIase activity

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Authors: Meenakshi Verma, Mandeep Singh Bakshi, Kultar Singh

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Magnetic nanoparticles have received incredible importance in view of their diverse applications, which arise primarily due to their response to the external magnetic field. The magnetic behaviour of magnetic nanoparticles (NPs) helps them in numerous different ways. The most important amongst them is the ease with which they can be purified and also can be separated from the media in which they are present merely by applying an external magnetic field. This exceptional ease of separation of the magnetic NPs from an aqueous media enables them to use for extracting/removing metal pollutants from complex aqueous medium. Functionalized magnetic NPs can be subjected for the metallic impurities extraction if are favourably adsorbed on the NPs surfaces. We have successfully used the magnetic NPs as vehicles for gold and silver NPs removal from the complex fluids. The NPs loaded with gold and silver NPs pollutant fractions has been easily removed from the aqueous media by using external magnetic field. Similarly, we have used the magnetic NPs for extraction of protein from complex media and then constantly washed with pure water to eliminate the unwanted surface adsorbed components for quantitative estimation. The purified and protein loaded magnetic NPs are best analyzed with SDS Page to not only for characterization but also for separating the protein fractions. A collective review of the results indicates that we have synthesized surfactant coated iron oxide NPs and then functionalized these with selected materials. These surface active magnetic NPs work very well for the extraction of metallic NPs from the aqueous bulk and make the whole process environmentally sustainable. Also, magnetic NPs-Au/Ag/Pd hybrids have excellent protein extracting properties. They are much easier to use in order to extract the magnetic impurities as well as protein fractions under the effect of external magnetic field without any complex conventional purification methods.

Keywords: magnetic nanoparticles, protein, functionalized, extraction

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Authors: Yasir Ali Arfat

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Composite films based on fish protein isolate (FPI) and fish skin gelatin (FSG) blend incorporated with 50 and 100% (w/w, protein) basil leaf essential oil (BEO) in the absence and presence of 3% (w/w, protein) ZnO nanoparticles (ZnONP) were prepared and characterised. Tensile strength (TS) decreased, whilst elongation at break (EAB) increased as BEO level increased (p < 0.05). However, ZnONP addition resulted in higher TS but lower EAB (p < 0.05). The lowest water vapour permeability (WVP) was observed for the film incorporated with 100% BEO and 3% ZnONP (p < 0.05). BEO and ZnONP incorporation decreased transparency of FPI/FSG films (p < 0.05). FTIR spectra indicated that films added with BEO exhibited higher hydrophobicity. Both BEO and ZnONP had a marked impact on thermal stability of the films. Microstructural study revealed that presence of ZnONP prevented bilayer formation of film containing 100% BEO. FPI/FSG films incorporated with 100% BEO, especially in combination with ZnONP, exhibited strong antibacterial activity against food pathogenic and spoilage bacteria and thus could be used as an active food packaging material to ensure safety and to extend the shelf-life of packaged foods.

Keywords: bionanocomposite, fish protein isolate, fish skin gelatin, basil essential oil, ZnO nanoparticles, antimicrobial packaging

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Authors: Muhammad Ajmal

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- Thermal processing and subsequent storage of ultra-heat treated (UHT) milk leads to alteration in protein profile, Maillard reaction and lipid oxidation. Concentration of carbohydrates in normal and flavored version of UHT milk is considerably different. Transition in protein profile, Maillard reaction and lipid oxidation in UHT flavored milk was determined for 90 days at ambient conditions and analyzed at 0, 45 and 90 days of storage. Protein profile, hydroxymethyl furfural, furosine, Nε-carboxymethyl-l-lysine, fatty acid profile, free fatty acids, peroxide value and sensory characteristics were determined. After 90 days of storage, fat, protein, total solids contents and pH were significantly less than the initial values determined at 0 day. As compared to protein profile normal UHT milk, more pronounced changes were recorded in different fractions of protein in UHT milk at 45 and 90 days of storage. Tyrosine content of flavored UHT milk at 0, 45 and 90 days of storage were 3.5, 6.9 and 15.2 µg tyrosine/ml. After 45 days of storage, the decline in αs1-casein, αs2-casein, β-casein, κ-casein, β-lactoglobulin, α-lactalbumin, immunoglobulin and bovine serum albumin were 3.35%, 10.5%, 7.89%, 18.8%, 53.6%, 20.1%, 26.9 and 37.5%. After 90 days of storage, the decline in αs1-casein, αs2-casein, β-casein, κ-casein, β-lactoglobulin, α-lactalbumin, immunoglobulin and bovine serum albumin were 11.2%, 34.8%, 14.3%, 33.9%, 56.9%, 24.8%, 36.5% and 43.1%. Hydroxy methyl furfural content of UHT milk at 0, 45 and 90 days of storage were 1.56, 4.18 and 7.61 (µmol/L). Furosine content of flavored UHT milk at 0, 45 and 90 days of storage intervals were 278, 392 and 561 mg/100g protein. Nε-carboxymethyl-l-lysine content of UHT flavored milk at 0, 45 and 90 days of storage were 67, 135 and 343mg/kg protein. After 90 days of storage of flavored UHT milk, the loss of unsaturated fatty acids 45.7% from the initial values. At 0, 45 and 90 days of storage, free fatty acids of flavored UHT milk were 0.08%, 0.11% and 0.16% (p<0.05). Peroxide value of flavored UHT milk at 0, 45 and 90 days of storage was 0.22, 0.65 and 2.88 (MeqO²/kg). Sensory analysis of flavored UHT milk after 90 days indicated that appearance, flavor and mouth feel score significantly decreased from the initial values recorded at 0 day. Findings of this investigation evidenced that in flavored UHT milk more pronounced changes take place in protein profile, Maillard reaction products and lipid oxidation as compared to normal UHT milk.

Keywords: UHT flavored milk , hydroxymethyl furfural, lipid oxidation, sensory properties

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Authors: Hwang Ahreum, Kang Taejoon, Kim Bongsoo

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Nanosensors for high sensitive detection of diseases have been widely studied to improve the quality of life. Here, we suggest robust nano-plasmonic diagnostic sensor using cysteine tagged protein G (Cys3-protein G) and ultraflat, ultraclean and single-crystalline Au nanoplates. Protein G formed on an ultraflat Au surface provides ideal background for dense and uniform immobilization of antibodies. The Au is highly stable in diverse biochemical environment and can immobilize antibodies easily through Au-S bonding, having been widely used for various biosensing applications. Especially, atomically smooth single-crystalline Au nanomaterials synthesized using chemical vapor transport (CVT) method are very suitable to fabricate reproducible sensitive sensors. As the C-reactive protein (CRP) is a nonspecific biomarker of inflammation and infection, it can be used as a predictive or prognostic marker for various cardiovascular diseases. Cys3-protein G immobilized uniformly on the Au nanoplate enable CRP antibody (anti-CRP) to be ordered in a correct orientation, making their binding capacity be maximized for CRP detection. Immobilization condition for the Cys3-protein G and anti-CRP on the Au nanoplate is optimized visually by AFM analysis. Au nanoparticle - Au nanoplate (NPs-on-Au nanoplate) assembly fabricated from sandwich immunoassay for CRP can reduce zero-signal extremely caused by nonspecific bindings, providing a distinct surface-enhanced Raman scattering (SERS) enhancement still in 10-18 M of CRP concentration. Moreover, the NP-on-Au nanoplate sensor shows an excellent selectivity against non-target proteins with high concentration. In addition, comparing with control experiments employing a Au film fabricated by e-beam assisted deposition and linker molecule, we validate clearly contribution of the Au nanoplate for the attomolar sensitive detection of CRP. We expect that the devised platform employing the complex of single-crystalline Au nanoplates and Cys3-protein G can be applied for detection of many other cancer biomarkers.

Keywords: Au nanoplate, biomarker, diagnostic sensor, protein G, SERS

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Authors: Debbarma Asis, Guha Chandan

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Tumor Protein-53 (P53) is one of the tumor suppressor proteins. P53 regulates the cell cycle that conserves stability by preventing genome mutation. It is named so as it runs as 53-kilodalton (kDa) protein on Polyacrylamide gel electrophoresis although the actual mass is 43.7 kDa. Experimental evidence has indicated that P53 cancer mutants loses tumor suppression activity and subsequently gain oncogenic activities to promote tumourigenesis. Tumor-specific DNA has recently been detected in the plasma of breast cancer patients. Detection of tumor-specific genetic materials in cancer patients may provide a unique and valuable tumor marker for diagnosis and prognosis. Commercially available MDA-468 breast cancer cell line was used for the proposed study.

Keywords: tumor protein (P53), cancer mutants, MDA-468, tumor suppressor gene

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Authors: Yazun Jarrar

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Smoking has effect on platelet aggregation and the activity of anti-platelet drugs. The chemical 20-hydroxyeicosatetraenoic acid (20-HETE) is a cardiotoxic arachidonic acid metabolite which increases platelet aggregation. In this study, we investigated the influence of cigarette smoking on 20-HETE levels and protein expression of 20-HETE producing enzyme CYP4A11 in isolated platelets from smoker and non-smoker volunteers. The protein expression and 20-HETE levels were analyzed using immunoblot and High-Performance Liquid Chromatography with Mass Spectrometry (HPL-MS) assays. The results showed that 20-HETE level was higher significantly among smokers than non-smokers (t-test, p-value<0.05). The protein expression of CYP4A11 was significantly higher (t-test, p-value<0.05) among the platelets of smokers. We concluded that cigarette smoking increased the level of platelet activator 20-HETE through increasing the protein expression of CYP4A11. These findings may increase the understanding of smoking-drug interaction during antiplatelets therapy.

Keywords: smoking, 20-HETE, CYP4A11, platelet

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Authors: Aghasadeghi MR., Bahramali G., Sadat SM., Sadeghi SA., Yousefi M., Khodaei K., Ghorbani M., Sadat Larijani M.

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Background:The emerging COVID-19 pandemic is a serious concernfor the public health worldwide. Despite the many mutations in the virus genome, it is important to find an effective vaccine against viral mutations. Therefore, in current study, we aimed at immunoinformatic evaluation of the virus proteins immunogenicity to design a preventive vaccine candidate, which could elicit humoral and cellular immune responses as well. Methods:Three antigenic regions are included;Spike, Membrane, and Nucleocapsid amino acid sequences were obtained, and possible fusion proteins were assessed andcompared by immunogenicity, structural features, and population coverage. The best fusion protein was also evaluated for MHC-I and MHC-II T-cell epitopes and the linear and conformational B-cell epitopes. Results: Among the four predicted models, the truncated Spike protein in fusion with M and N proteins is composed of 24 highly immunogenic human MHC class I and 29 MHC class II, along with 14 B-cell linear and 61 discontinues epitopes. Also, the selected protein has high antigenicity and acceptable population coverage of 82.95% in Iran and 92.51% in Europe. Conclusion: The data indicate that the truncated Spike-M-N SARS-CoV-2form which could be potential targets of neutralizing antibodies. The protein also has the ability to stimulate humoral and cellular immunity. The in silico study provided the fusion protein as a potential preventive vaccine candidate for further in vivo evaluation.

Keywords: SARS-CoV-2, immunoinformatic, protein, vaccine

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Authors: Kah Yan How, Peh Fern Ong, Keang Peng Song

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Porphyromonas gingivalis is a black-pigmented, anaerobic Gram-negative bacterium that is important in the progression of chronic and severe periodontitis. This organism has an essential requirement for iron, which is usually obtained from hemin, using specific membrane receptors, proteases, and lipoproteins. In this study, we report the characterization of a novel 24 kDa hemin-binding protein, HmuX, in P. gingivalis W50. The hmuX gene is 651 bp long which encodes for a 217 amino acid protein. HmuX was found to be identical at the C-terminus to the previously reported HmuY protein, differing by an additional 74 amino acids at the N-terminus. Recombinant HmuX demonstrated hemin-binding ability by LDS- PAGE and TMBZ staining. Sequence analysis of HmuX revealed a putative lipoprotein attachment site, suggesting its possible role as a lipoprotein. HmuX was also localized to the outer cell surface by transmission electron microscopy. Northern analysis showed hmuX to be transcribed as a single gene and that hmuX mRNA was tightly regulated by the availability of extra-cellular hemin. P. gingivalis isogenic mutant deficient in hmuX gene exhibited significant growth retardation under hemin-limited conditions. Taken together, these results suggest that HmuX is a hemin-binding lipoprotein, important in hemin utilization for the growth of P. gingivalis.

Keywords: Porphyromonas gingivalis, periodontal diseases, HmuX, protein characterization

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Authors: Hyun-Woo Jun, Tae-Chul Jeong, Huai-Cong Liu, Ju Lee

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The paper presents the electromagnetic-thermal flow coupled analysis of permanent magnet synchronous motor (PMSM) which has cooling channel in stator core for forced air cooling. Unlike the general PMSM design, to achieve ohmic loss reduction for high efficiency, cooling channel actively used in the stator core. Equivalent thermal network model was made to analyze the effect of the formation of the additional flow path in the core. According to the shape and position changing of the channel design, electromagnetic-thermal coupled analysis results were reviewed.

Keywords: coupled problems, electric motors, equivalent circuits, fluid flow, thermal analysis

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Authors: Che Hsiu Chen, Su Yun Chen, Hon Wen Cheng

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Traditional core stability, core endurance, and balance exercises on a stable surface with isometric muscle actions, low loads, and multiple repetitions, which may not improvements the swimming and running economy performance. However, the effects of high intensity core muscle exercise training on jump height, sprint, and aerobic fitness remain unclear. The purpose of this study was to examine whether high intensity core muscle exercises training could improve sport performances in dancers. Thirty healthy university dancer students (28 women and 2 men; age 20.0 years, height 159.4 cm, body mass 52.7 kg) were voluntarily participated in this study, and each participant underwent five suspension exercises (e.g., hip abduction in plank alternative, hamstring curl, 45-degree row, lunge and oblique crunch). Each type of exercise was performed for 30-second, with 30-second of rest between exercises, two times per week for eight weeks and each exercise session was increased by 10-second every week. We measured agility, explosive force, anaerobic and cardiovascular fitness in dancer performance before and after eight weeks of training. The results showed that the 8-week high intensity core muscle training would significantly increase T-test agility (7.78%), explosive force of acceleration (3.35%), vertical jump height (8.10%), jump power (6.95%), lower extremity anaerobic ability (7.10%) and oxygen uptake efficiency slope (4.15%). Therefore, it can be concluded that eight weeks of high intensity core muscle exercises training can improve not only agility, sprint ability, vertical jump ability, anaerobic and but also cardiovascular fitness measures as well.

Keywords: balance, jump height, sprint, maximal oxygen uptake

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Authors: Oscar Rubiano, Oscar Ortiz, Natalia Morales, Lida Alfonso, Johana Alvarado, Adriana Gutierrez, Daniel Botero

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Athletes who usually use protein supplements, are those who practice strength and power sports, whose goal is to achieve a large muscle mass. However, it has also been explored in sports or endurance activities such as cycling, and where despite requiring high power, prominent muscle development can impede good competitive performance due to the determinant of body mass for good performance of the athlete body. This research shows, the effect with protein supplements establishes a protein - muscle mass ratio, although in a lesser proportion the relationship between protein types and muscle power. Thus, we intend to explore as a first approximation, the behavior of muscle power in lower limbs after the intake of two protein supplements from different sources. The aim of the study was to describe the behavior of muscle power in lower limbs after the consumption of animal protein (AP) and vegetable protein (VP) in four route cyclists from 18 to 20 years of the Bogota cycling league. The methodological design of this study is quantitative, with a non-probabilistic sampling, based on a pre-experimental model. The jumping power was evaluated before and after the intervention by means of the squat jump test (SJ), Counter movement jump (CMJ) and Abalacov (AB). Cyclists consumed a drink with whey protein and a soy isolate after training four times a week for three months. The amount of protein in each cyclist, was calculated according to body weight (0.5 g / kg of muscle mass). The results show that subjects who consumed PV improved muscle strength and landing strength. In contrast, the power and landing force decreased for subjects who consumed PA. For the group that consumed PV, the increase was positive at 164.26 watts, 135.70 watts and 33.96 watts for the AB, SJ and CMJ jumps respectively. While for PA, the differences of the medians were negative at -32.29 watts, -82.79 watts and -143.86 watts for the AB, SJ and CMJ jumps respectively. The differences of the medians in the AB jump were positive for both the PV (121.61 Newton) and PA (454.34 Newton) cases, however, the difference was greater for PA. For the SJ jump, the difference for the PA cases was 371.52 Newton, while for the PV cases the difference was negative -448.56 Newton, so the difference was greater in the SJ jump for PA. In jump CMJ, the differences of the medians were negative for the cases of PA and PV, being -7.05 for PA and - 958.2 for PV. So the difference was greater for PA. The conclusion of this study shows that serum protein supplementation showed no improvement in muscle power in the lower limbs of the cyclists studied, which could suggest that whey protein does not have a beneficial effect on performance in terms of power, either, showed an impact on body composition. In contrast, supplementation with soy isolate showed positive effects on muscle power, body.

Keywords: animal protein (AP), muscle power, supplements, vegetable protein (VP)

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Authors: F. C. Amadi, G. C. Enyi, G. Nasr

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Relative permeabilities are practical factors that are used to correct the single phase Darcy’s law for application to multiphase flow. For effective characterisation of large-scale multiphase flow in hydrocarbon recovery, relative permeability and capillary pressures are used. These parameters are acquired via special core flooding experiments. Special core analysis (SCAL) module of reservoir simulation is applied by engineers for the evaluation of these parameters. But, core flooding experiments in shale core sample are expensive and time consuming before various flow assumptions are achieved for instance Darcy’s law. This makes it imperative for the application of coreflooding simulations in which various analysis of relative permeabilities and capillary pressures of multiphase flow can be carried out efficiently and effectively at a relative pace. This paper presents a Sendra software simulation of core flooding to achieve to relative permeabilities and capillary pressures using different correlations. The approach used in this study was three steps. The first step, the basic petrophysical parameters of Marcellus shale sample such as porosity was determined using laboratory techniques. Secondly, core flooding was simulated for particular scenario of injection using different correlations. And thirdly the best fit correlations for the estimation of relative permeability and capillary pressure was obtained. This research approach saves cost and time and very reliable in the computation of relative permeability and capillary pressures at steady or unsteady state, drainage or imbibition processes in oil and gas industry when compared to other methods.

Keywords: relative permeabilty, porosity, 1-D black oil simulator, capillary pressures

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Authors: Hamid Hossein Khezri, Afsaneh Javdani-Mallak

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Fast inhibitory neurotransmission in the mammalian nervous system is largely mediated by GABAA receptors, chloride-selective members of the superfamily of pentameric Cys-loop receptors. Cannabidiol (CBD) is one of the members of cannabinoid compounds found in cannabis. CBD and Cannabinol (CBN), as the other extract of plant Cannabis were able to reduce myofascial pain in rats with immunosuppressive and anti-inflammatory activities. In this study, we accomplished protein-protein BLAST, and the sequence was found to be for Gamma-aminobutyric acid receptor subunit alpha-1 (GBRA1) chain A and its 3D structure was subsequently downloaded from Protein Data Bank. The structures of the ligands, cannabinol, and cannabidiol, were obtained from PubChem. After the necessary process of the obtained files, AutoDock Vina was used to perform molecular docking. Docking between the ligands and GBRA1 chain A revealed that cannabinol has a higher affinity to GBRA1 (binding energy = -7.5 kcal/mol) compared to cannabidiol (binding energy = -6.5 kcal/mol). Furthermore, cannabinol seems to be able to interact with 10 residues of the protein, out of which 3 are in the neurotransmitter-gated ion-channel transmembrane domain of GBRA1, whereas cannabidiol interacts with two other residues. Although the results of this project do not indicate the activating /or inhibitory capability of the studied compounds, it suggests that cannabinol can act as a relatively strong ligand for GBRA1.

Keywords: protein-ligand docking, cannabinol, cannabidiol, GBRA1

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Authors: Zahra Akbari, Farzin Zokaee, Talat Ghomashchi

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Whey is a by-product of the dairy industry whose major components are lactose (44–52 g/L), proteins (6–8 g/L) and mineral salts (4–9 g/L). Approximately 50% of 121 million tons of whey produced in the world in 1993 were disposed into rivers, lakes or other water bodies, treated in wastewater treatment plants or loaded onto land. This represents a significant loss of resources and causes serious pollution problems since whey is a heavy organic pollutant with high COD and BOD values, 40–60 g/L and 50–80 g/L, respectively. The removal of cheese whey proteins and minerals represent an important task both in environmental and in food sciences. The most important treatments which are considered in this study, have been done by using lime, Al2O3, FeCl3 and AlCl3 along with heating and also acidic-alkaline method. Results show that the best way for removal of protein is accomplished with adding HCl to decrease pH from 6 to 4, boiling for 20 min, and filtering protein aggregates. Also partial demineralization in whey solution for reducing ash is accomplished by adding NaOH to increase pH to 7.2 and heating solution for 20 min.

Keywords: whey treatment, dairy industry, precipitation, protein, mineral

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Authors: Shahram Naghizadeh Raeisi, Ali Alghooneh

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The application of whey protein in drink industry to enhance the nutritional value of the products is important. Furthermore, the gelification of protein during thermal treatment and shelf life makes some limitations in its application. So, the main goal of this research is manufacturing of high concentrate whey protein orange drink with appropriate shelf life. In this way, whey protein was 5 to 30% hydrolyzed ( in 5 percent intervals at six stages), then thermal stability of samples with 10% concentration of protein was tested in acidic condition (T= 90 °C, pH=4.2, 5 minutes ) and neutral condition (T=120° C, pH:6.7, 20 minutes.) Furthermore, to study the shelf life of heat treated samples in 4 months at 4 and 24 °C, the time sweep rheological test were done. At neutral conditions, 5 to 20% hydrolyzed sample showed gelling during thermal treatment, whereas at acidic condition, was happened only in 5 to 10 percent hydrolyzed samples. This phenomenon could be related to the difference in hydrodynamic radius and zeta potential of samples with different level of hydrolyzation at acidic and neutral conditions. To study the gelification of heat resistant protein solutions during shelf life, for 4 months with 7 days intervals, the time sweep analysis were performed. Cross over was observed for all heat resistant neutral samples at both storage temperature, while in heat resistant acidic samples with degree of hydrolysis, 25 and 30 percentage at 4 and 20 °C, it was not seen. It could be concluded that the former sample was stable during heat treatment and 4 months storage, which made them a good choice for manufacturing high protein drinks. The Scheffe polynomial model and numerical optimization were employed for modeling and high protein orange drink formula optimization. Scheffe model significantly predicted the overal acceptance index (Pvalue<0.05) of sensorial analysis. The coefficient of determination (R2) of 0.94, the adjusted coefficient of determination (R2Adj) of 0.90, insignificance of the lack-of-fit test and F value of 64.21 showed the accuracy of the model. Moreover, the coefficient of variable (C.V) was 6.8% which suggested the replicability of the experimental data. The desirability function had been achieved to be 0.89, which indicates the high accuracy of optimization. The optimum formulation was found as following: Modified whey protein solution (65.30%), natural orange juice (33.50%), stevia sweetener (0.05%), orange peel oil (0.15%) and citric acid (1 %), respectively. Its worth mentioning that this study made an appropriate model for application of whey protein in drink industry without bitter flavor and gelification during heat treatment and shelf life.

Keywords: croos over, orange beverage, protein modification, optimization

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Authors: Santhosh Nallapu, G. Radhakrishnamacharya

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A two-fluid model of Herschel-Bulkley fluid flow through tubes of small diameters is studied. It is assumed that the core region consists of Herschel-Bulkley fluid and Newtonian fluid in the peripheral region. The analytical solutions for velocity, flow flux, effective viscosity, core hematocrit and mean hematocrit have been derived and the effects of various relevant parameters on these flow variables have been studied. It has been observed that the effective viscosity and mean hematocrit increase with yield stress, power-law index, hematocrit and tube radius. Further, the core hematocrit decreases with hematocrit and tube radius.

Keywords: two-layered model, non-Newtonian fluid, hematocrit, Fahraeus-Lindqvist effect, plug flow

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Authors: Avinoam Rabinovich

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CO₂ geological storage is a proven technology for reducing anthropogenic carbon emissions, which is paramount for achieving the ambitious net zero emissions goal. Core flooding experiments are an important step in any CO₂ storage project, allowing us to gain information on the flow of CO₂ and brine in the porous rock extracted from the reservoir. This information is important for understanding basic mechanisms related to CO₂ geological storage as well as for reservoir modeling, which is an integral part of a field project. In this work, a different method for constructing accurate models of CO₂-brine core flooding will be presented. Results for synthetic cases and real experiments will be shown and compared with numerical models to exhibit their predictive capabilities. Furthermore, the various mechanisms which impact the CO₂ distribution and trapping in the rock samples will be discussed, and examples from models and experiments will be provided. The new method entails solving an inverse problem to obtain a three-dimensional permeability distribution which, along with the relative permeability and capillary pressure functions, constitutes a model of the flow experiments. The model is more accurate when data from a number of experiments are combined to solve the inverse problem. This model can then be used to test various other injection flow rates and fluid fractions which have not been tested in experiments. The models can also be used to bridge the gap between small-scale capillary heterogeneity effects (sub-core and core scale) and large-scale (reservoir scale) effects, known as the upscaling problem.

Keywords: CO₂ geological storage, residual trapping, capillary heterogeneity, core flooding, CO₂-brine flow

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Authors: Manssour Habbash, Srinivasa Rao Idapalapati

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Although writing is a multifaceted task, teaching writing is a demanding task basically for two reasons: Grammar and Syntax. This article provides a method of teaching writing that was found to be effective in improving students’ academic writing composition skill. The article explains the concepts of ‘guided-discovery’ and ‘guided-construction’ upon which a method of teaching writing is grounded and developed. Providing a brief commentary on what the core could mean primarily, the article presents an exposition of understanding and identifying the core and building upon the core that can demonstrate the way a teacher can make use of the concepts in teaching for improving the writing skills of their students. The method is an adaptation of grammar translation method that has been improvised to suit to a student-centered classroom environment. An intervention of teaching writing through this method was tried out with positive outcomes in formal classroom research setup, and in view of the content’s quality that relates more to the classroom practices and also in consideration of its usefulness to the practicing teachers the process and the findings are presented in a narrative form along with the results in tabular form.

Keywords: core of a text, guided construction, guided discovery, theme of a text

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Authors: Berhan Fikru

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The objective of this study was to develop new complementary food from maize, soybean and moringa for young children. The complementary foods were formulated with linear programming (LP Nutri-survey software) and Faffa (corn soya blend) use as control. Analysis were made for formulated blends and compared with the control and recommended daily intake (RDI). Three complementary foods composed of maize, soya bean, moringa and sugar with ratio of 65:20:15:0, 55:25:15:5 and 65:20:10:5 for blend 1, 2 and 3, respectively. The blends were formulated based on the protein, energy, mineral (iron, zinc an calcium) and vitamin (vitamin A and C) content of foods. The overall results indicated that nutrient content of faffa (control) was 16.32 % protein, 422.31 kcal energy, 64.47 mg calcium, 3.8 mg iron, 1.87mg zinc, 0.19 mg vitamin A and 1.19 vitamin C; blend 1 had 17.16 % protein, 429.84 kcal energy, 330.40 mg calcium, 6.19 mg iron, 1.62 mg zinc, 6.33 mg vitamin A and 4.05 mg vitamin C; blend 2 had 20.26 % protein, 418.79 kcal energy, 417.44 mg calcium, 9.26 mg iron, 2.16 mg zinc, 8.43 mg vitamin A and 4.19 mg vitamin C whereas blend 3 exhibited 16.44 % protein, 417.42 kcal energy, 242.4 mg calcium, 7.09 mg iron, 2.22 mg zinc, 3.69 mg vitamin A and 4.72 mg vitamin C, respectively. The difference was found between all means statically significance (P < 0.05). Sensory evaluation showed that the faffa control and blend 3 were preferred by semi-trained panelists. Blend 3 had better in terms of its mineral and vitamin content than FAFFA corn soya blend and comparable with WFP proprietary products CSB+, CSB++ and fulfills the WHO recommendation for protein, energy and calcium. The suggested formulation with Moringa powder can therefore be used as a complementary food to improve the nutritional status and also help solve problems associated with protein energy and micronutrient malnutrition for young children in developing countries, particularly in Ethiopia.

Keywords: corn soya blend, proximate composition, micronutrient, mineral chelating agents, complementary foods

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Authors: M. Chaichanyut, S. Tungjitkusolmun

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This paper is concerned with microwave (MW) ablation for a liver cancer tissue by using helix antenna. The antenna structure supports the propagation of microwave energy at 2.45 GHz. A 1½ turn spiral catheter-based microwave antenna applicator has been developed. We utilize the three-dimensional finite element method (3D FEM) simulation to analyze where the tissue heat flux, lesion pattern and volume destruction during MW ablation. The configurations of helix antenna where Helix air-core antenna and Helix Dielectric-core antenna. The 3D FEMs solutions were based on Maxwell and bio-heat equations. The simulation protocol was power control (10 W, 300s). Our simulation result, both helix antennas have heat flux occurred around the helix antenna and that can be induced the temperature distribution similar (teardrop). The region where the temperature exceeds 50°C the microwave ablation was successful (i.e. complete destruction). The Helix air-core antenna and Helix Dielectric-core antenna, ablation zone or axial ratios (Widest/length) were respectively 0.82 and 0.85; the complete destructions were respectively 4.18 cm³ and 5.64 cm³.

Keywords: liver cancer, Helix antenna, finite element, microwave ablation

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Authors: Fengmei Li, Li Xu, Guoliang Xia

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Whey acidic proteins (WAP) domain-containing proteins in crustacean are involved in innate immune response against microbial invasion. In the present study, a novel double WAP domain (DWD)-containing protein gene 3 was identified from Chinese mitten crab Eriocheir sinensis (designated EsDWD3) by expressed sequence tag (EST) analysis and PCR techniques. The full-length cDNA of EsDWD3 was of 1223 bp, consisting of a 5′-terminal untranslated region (UTR) of 74 bp, a 3′ UTR of 727 bp with a polyadenylation signal sequence AATAAA and a polyA tail, and an open reading frame (ORF) of 423 bp. The ORF encoded a polypeptide of 140 amino acids with a signal peptide of 22 amino acids. The deduced protein sequence EsDWD3 showed 96.4 % amino acid similar to other reported EsDWD1 from E. sinensis, and phylogenetic tree analysis revealed that EsDWD3 had closer relationships with the reported two double WAP domain-containing proteins of E. sinensis species.

Keywords: Chinese mitten crab, Eriocheir sinensis, cloning, double WAP domain-containing protein

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Authors: Hamid Hossein Khezri, Afsaneh Javdani-Mallak

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Fast inhibitory neurotransmission in the mammalian nervous system is largely mediated by GABAA receptors, chloride-selective members of the superfamily of pentameric Cys-loop receptors. Cannabidiol (CBD) is one of the members of cannabinoid compounds found in cannabis. CBD and Cannabinol (CBN), as the other extract of plant Cannabis, were able to reduce myofascial pain in rats with immunosuppressive and anti-inflammatory activities. In this study, we accomplished protein-protein BLAST and the sequence was found to be for Gamma-aminobutyric acid receptor subunit alpha-1 (GBRA1) chain A and its 3D structure was subsequently downloaded from Protein Data Bank. The structures of the ligands cannabinol and cannabidiol were obtained from PubChem. After a necessary process of the obtained files, AutoDock Vina was used to performing molecular docking. Docking between the ligands and GBRA1 chain A revealed that cannabinol has a higher affinity to GBRA1 (binding energy = -7.5 kcal/mol) compared to cannabidiol (binding energy = -6.5 kcal/mol). Furthermore, cannabinol seems to be able to interact with 10 residues of the protein, out of which 3 are in the neurotransmitter-gated ion-channel transmembrane domain of GBRA1, whereas cannabidiol interacts with two other residues. Although the results of this project do not indicate the activating /or inhibitory capability of the studied compounds, it suggests that cannabinol can act as a relatively strong ligand for GBRA1.

Keywords: protein-ligand docking, cannabinol, cannabidiol, GBRA1

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Authors: Keaghan Brown

Abstract:

The prioritization of drug resistance mutations impacting protein folding or protein-drug and protein-DNA interactions within macromolecular systems is critical to the success of treatment regimens. With a continual increase in computational tools to assess these impacts, the need for scalability and reproducibility became an essential component of computational analysis and experimental research. Here it introduce a bioinformatics pipeline that combines several structural analysis tools in a simplified workflow, by optimizing the present computational hardware and software to automatically ease the flow of data transformations. Utilizing preestablished software tools, it was possible to develop a pipeline with a set of pre-defined functions that will automate mutation introduction into the HIV-1 Integrase protein structure, calculate the gain and loss of polar interactions and calculate the change in energy of protein fold. Additionally, an automated molecular dynamics analysis was implemented which reduces the constant need for user input and output management. The resulting pipeline, Automated Mutation Introduction and Analysis (AMIA) is an open source set of scripts designed to introduce and analyse the effects of mutations on the static protein structure as well as the results of the multi-conformational states from molecular dynamic simulations. The workflow allows the user to visualize all outputs in a user friendly manner thereby successfully enabling the prioritization of variant systems for experimental validation.

Keywords: automated workflow, variant prioritization, drug resistance, HIV Integrase

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Authors: Tzyy-Rong Jinn, Sheng-Kuo Hsieh, Yi-Ching Chung, Feng-Chia Hsieh

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In this study, we attempted to develop a recombinant oleosin-based fusion expression strategy in Escherichia coli (E. coli) and coupled with the artificial oil bodies (AOB)-cyanogen bromide technology platform to produce bioactive mastoparan B (MP-B). As reported, the oleosin in AOB system plays a carrier (fusion with target protein), since oleosin possess two amphipathic regions (at the N-terminus and C-terminus), which result in the N-terminus and C-terminus of oleosin could be arranged on the surface of AOB. Thus, the target protein fused to the N-terminus or C-terminus of oleosin which also is exposed on the surface of AOB, and this process will greatly facilitate the subsequent separation and purification of target protein from AOB. In addition, oleosin, a unique structural protein of seed oil bodies, has the added advantage of helping the fused MP-B expressed in inclusion bodies, which can protect from proteolytic degradation. In this work, MP-B was fused to the C-terminus of oleosin and then was expressed in E. coli as an insoluble recombinant protein. As a consequence, we successfully developed a reliable recombinant oleosin-based fusion expression strategy in Escherichia coli and coupled with the artificial oil bodies (AOB)-cyanogen bromide technology platform to produce the small peptide, MP-B. Take together, this platform provides an insight into the production of active MP-B, which will facilitate studies and applications of this peptide in the future.

Keywords: artificial oil bodies, Escherichia coli, Oleosin-fusion protein, Mastoparan-B

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Authors: Zyrah Lou R. Samar, Genecarlo Liwanag

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Type 2 Diabetes Mellitus is the more prevalent type, caused by a combination of insulin resistance and inadequate insulin response to hyperglycemia1. Aside from pharmacologic interventions, medical nutrition therapy is an integral part of the management of patients with Type 2 Diabetes Mellitus. Whey protein, which is one of the best protein sources, has been investigated for its applicability in improving glycemic control in patients with Type 2 Diabetes Mellitus. This systematic review and meta-analysis was conducted to measure the magnitude of the effect of whey protein on glycemic control in type 2 diabetes mellitus. The aim of this review is to evaluate the efficacy and safety of whey protein in patients with type 2 diabetes mellitus. Methods: A systematic electronic search for studies in the PubMed and Cochrane Collaboration database was done. Included in this review were randomized controlled trials of whey protein enrolling patients with type 2 diabetes mellitus. Three reviewers independently searched, assessed, and extracted data from the individual studies. Results: A systematic literature search on online databases such as Cochrane Central Registry, PubMed, and Herdin Plus was conducted in April to September 2021 to identify eligible studies. The search yielded 21 randomized controlled trials after removing duplicates. Only 5 articles were included after reviewing the full text, which met the criteria for selection. Conclusion: Whey protein supplementation significantly reduced fasting blood glucose. However, it did not reduce post-prandial blood glucose, HbA1c level, and weight when compared with the placebo. There has been a considerate heterogeneity across all studies, which may have contributed/confounded its effects. A larger sample size and better inclusion, and a more specific study may be included in the future reviews.

Keywords: whey protein, diabetes, nutrition, fasting blood sugar, postprandial glucose, HbA1c, weight reduction

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Authors: Khansa Hayat

Abstract:

Organizational stress is one of the vital phenomena which is having its roots deep down in has deep roots in management, psychology, and organizational behavior research. In the meanwhile, keeping its focus on the positive strength of humans rather than the traditional negativity oriented research, positive psychology has emerged as a separate branch of organizational behavior. The current study investigates the interactive effects of Challenge and hindrance stressors and core Self Evaluations (CSE’s) of the individual on job performances including the in-role performance and extra role performances. The study also aims to investigate the supporting/buffering role of the human dispositions (i.e., self esteem, self efficacy, locus of control and emotional stability). The results show that Challenge stressors have a significant positive effect on in role performance and extra role performance of the individual. The findings of the study indicate that Core Self evaluations strengthen the relationship between challenge stressors and in role performance of the individual. In case of Hindrance Stressors the Core self Evaluations lessen the negative impact of Hindrance stressors and they let the individual perform at a better and normal position even when the Hindrance stressors are high. The relationship and implication of conservation of resource theory are also discussed. The limitations, future research directions and implications of the study are also discussed.

Keywords: challenge-hindrance stressors, core self evaluations, in-role performance, extra-role performance

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Authors: Jiun-Nan Hou, Tien-Huang Chen, Wei-June Chen

Abstract:

Dengue viruses (DENVs) are naturally transmitted between humans by mosquito vectors. Mosquito cells usually survive DENV infection, allowing infected mosquitoes to retain an active status for virus transmission. In this study, we found that DENV2 virus infection in mosquito cells causes the unfolded protein response (UPR) that activates the protein kinase RNA-like endoplasmic reticulum kinase (PERK) signal pathway, leading to shutdown of global protein translation in infected cells which was apparently regulated by the PERK signal pathway. According to observation in this study, the PERK signal pathway in DENV2-infected C6/36 cells alleviates ER stress, and reduces initiator and effector caspases, as well as the apoptosis rate via shutdown of cellular proteins. In fact, phosphorylation of eukaryotic initiation factor 2ɑ (eIF2ɑ) by the PERK signal pathway may impair recruitment of ribosomes that bind to the mRNA 5’-cap structure, resulting in an inhibitory effect on canonical cap-dependent cellular protein translation. The resultant pro-survival “byproduct” of infected mosquito cells is undoubtedly advantageous for viral replication. This finding provides insights into elucidating the PERK-mediated modulating web that is actively involved in dynamic protein synthesis, cell survival, and viral replication in mosquito cells.

Keywords: cap-dependent protein translation, dengue virus, endoplasmic reticulum stress, mosquito cells, PERK signal pathway

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Authors: Nutnicha Wongpadungkiat, Suwit Siriwatanayotin, Aluck Thipayarat, Punchira Vongsawasdi, Chotika Viriyarattanasak

Abstract:

Chicken products are major export product of Thailand. With a dramatically increasing consumption of chicken product in the world, there are abundant wastes from chicken meat processing industry. Recently, much research in the development of value-added products from chicken meat industry has focused on the production of protein hydrolysate, utilized as food ingredients for human diet and animal feed. The present study aimed to determine the effect of thermal pre-treatment on functional properties of chicken protein hydrolysate. Chicken breasts were heated at 40, 60, 80 and 100ºC prior to hydrolysis by Alcalase at 60ºC, pH 8 for 4 hr. The hydrolysate was freeze-dried, and subsequently used for assessment of its functional properties molecular weight by gel electrophoresis (SDS-PAGE). The obtained results show that increasing the pre-treatment temperature increased oil holding capacity and emulsion stability while decreasing antioxidant activity and water holding capacity. The SDS-PAGE analysis showed the evidence of protein aggregation in the hydrolysate treated at the higher pre-treatment temperature. These results suggest the connection between molecular weight of the hydrolysate and its functional properties.

Keywords: chicken protein hydrolysate, enzymatic hydrolysis, thermal pretreatment, functional properties

Procedia PDF Downloads 245