Search results for: protein cage nanoparticles

Authors: Mohammed Omar

Abstract:

Using Chenopodium murale leaf, gold nanoparticles (Au NP's) were biosynthesized effectively in an amicable strategy. The casting process was used to create composite layers of sodium alginate and polyvinyl pyrrolidone. Gold nanoparticles were incorporated into the polyvinyl pyrrolidone (PVP)/ sodium alginate (NaAlg) polymer blend by casting technique. Before and after exposure to different doses of gamma irradiation (2, 4, 6 Mrad), thin films of synthesized nanocomposites were analyzed. XRD revealed the amorphous nature of polymer blends (PVP/ NaAlg), which decreased by both Au NP's embedding and consecutive doses of irradiation. FT-IR spectra revealed interactions and differences within the functional groups of their respective pristine components and dopant nano-fillers. The optical properties of PVP/NaAlg – Au NP thin films (refractive index n, energy gap Eg, Urbach energy Eu) were examined before and after the irradiation procedure. Transmission electron micrographs (TEM) demonstrated a decrease in the size of Au NP’s and narrow size distribution as the gamma irradiation dose was increased. Gamma irradiation was found to influence the electrical conductivity of synthesized composite films, as well as dielectric permittivity (ɛ′) and dielectric losses (ε″).

Keywords: PVP, SPR, γ-radiations, XRD

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Authors: Rajeev Jain, Nimisha Jadon, Kshiti Singh

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Titanium oxide nanoparticles and 1-butyl-2,3-dimethylimidazolium bis (trifluoromethane- sulfonyl) imide modified glassy carbon electrode (TiO2/IL/GCE) has been fabricated for electrochemical sensing of flunarizine dihydrochloride (FRH). The electrochemical properties and morphology of the prepared nanocomposite were studied by electrochemical impedance spectroscopy (EIS) and transmission electron microscopy (TEM). The response of the electrochemical sensor was found to be proportional to the concentrations of FRH in the range from 0.5 µg mL-1 to 16 µg mL-1. The detection limit obtained was 0.03 µg mL-1. The proposed method was also applied to the determination of FRH in pharmaceutical formulation and human serum with good recoveries.

Keywords: flunarizine dihydrochloride, ionic liquid, nanoparticles, voltammetry, human serum

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Authors: Ganesh K. Maurya, Reema Chaudhary, Neha Pandey, Hari S. Misra

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PprA, a pleiotropic protein participating in radioresistance, has been reported for its roles in DNA replication initiation, genome segregation, cell division and DNA repair in polyextremophile Deinococcus radiodurans. Interestingly, expression of deinococcal PprA in E. coli suppresses its growth by reducing the number of colony forming units and provides better resistance against γ-radiation than control. We employed different biochemical and cell biology studies using PprA and its DNA binding/polymerization mutants (K133E & W183R) in E. coli. Cells expressing wild type PprA or its K133E mutant showed reduction in the amount of genomic DNA as well as chromosome copy number in comparison to W183R mutant of PprA and control cells, which suggests the role of PprA protein in regulation of DNA replication initiation in E. coli. Further, E. coli cells expressing PprA or its mutants exhibited different impact on cell morphology than control. Expression of PprA or K133E mutant displayed a significant increase in cell length upto 5 folds while W183R mutant showed cell length similar to uninduced control cells. We checked the interaction of deinococcal PprA and its mutants with E. coli DnaA using Bacterial two-hybrid system and co-immunoprecipitation. We observed a functional interaction of EcDnaA with PprA and K133E mutant but not with W183R mutant of PprA. Further, PprA or K133E mutant has suppressed the ATPase activity of EcDnaA but W183R mutant of PprA failed to do so. These observations suggested that PprA protein regulates DNA replication initiation and cell morphology of surrogate E. coli.

Keywords: DNA replication, radioresistance, protein-protein interaction, cell morphology, ATPase activity

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Authors: Naser Valipour Motlagh, Majid Aliabadi, Elnaz Rahmani, Samira Ghorbanpour

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Strawberry is one of the most favored fruits all along the world. But due to its vulnerability to microbial contamination and short life storage, there are lots of problems in industrial production and transportation of this fruit. Therefore, lots of ideas have tried to increase the storage life of strawberries especially through proper packaging. This paper works on efficient packaging as well. The primary material used is produced through simple mixing of low-density polyethylene (LDPE) and silver nanoparticles in different weight fractions of 0.5 and 1% in presence of dicumyl peroxide as a cross-linking agent. Final packages were made in a twin-screw extruder. Then, their effect on the quality maintenance of strawberry is evaluated. The SEM images of nano-silver packages show the distribution of silver nanoparticles in the packages. Total bacteria count, mold, yeast and E. coli are measured for microbial evaluation of all samples. Texture, color, appearance, odor, taste and total acceptance of various samples are evaluated by trained panelists and based on 9-point hedonic scale method. The results show a decrease in total bacteria count and mold in nano-silver packages compared to the samples packed in polyethylene packages for the same storage time. The optimum concentration of silver nanoparticles for the lowest bacteria count and mold is predicted to be around 0.5% which has attained the most acceptance from the panelist as well. Moreover, organoleptic properties of strawberry are preserved for a longer period in nano-silver packages. It can be concluded that using nano-silver particles in strawberry packages has improved the storage life and quality maintenance of the fruit.

Keywords: antimicrobial properties, polyethylene, silver nanoparticles, strawberry

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

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

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

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

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

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

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

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

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

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

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

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

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Authors: Ganesh K. Maurya, Reema Chaudhary, Neha Pandey, Hari S. Misra

Abstract:

PprA, a pleiotropic protein participating in radioresistance, has been reported for its roles in DNA replication initiation, genome segregation, cell division and DNA repair in polyextremophile Deinococcus radiodurans. Interestingly, expression of deinococcal PprA in E. coli suppresses its growth by reducing the number of colony forming units and provide better resistance against γ-radiation than control. We employed different biochemical and cell biology studies using PprA and its DNA binding/polymerization mutants (K133E & W183R) in E. coli. Cells expressing wild type PprA or its K133E mutant showed reduction in the amount of genomic DNA as well as chromosome copy number in comparison to W183R mutant of PprA and control cells, which suggests the role of PprA protein in regulation of DNA replication initiation in E. coli. Further, E. coli cells expressing PprA or its mutants exhibited different impact on cell morphology than control. Expression of PprA or K133E mutant displayed a significant increase in cell length upto 5 folds while W183R mutant showed cell length similar to uninduced control cells. We checked the interaction of deinococcal PprA and its mutants with E. coli DnaA using Bacterial two-hybrid system and co-immunoprecipitation. We observed a functional interaction of EcDnaA with PprA and K133E mutant but not with W183R mutant of PprA. Further, PprA or K133E mutant has suppressed the ATPase activity of EcDnaA but W183R mutant of PprA failed to do so. These observations suggested that PprA protein regulates DNA replication initiation and cell morphology of surrogate E. coli.

Keywords: DNA replication, radioresistance, protein-protein interaction, cell morphology, ATPase activity

Procedia PDF Downloads 45

Authors: Evelyn Osehontue Uroro, Richard Bright, Jing Yang Quek, Krasimir Vasilev

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Bacterial infections have been a challenge both in the public and private sectors. The colonization of bacteria often occurs in medical devices such as catheters, heart valves, respirators, and orthopaedic implants. When biomedical devices are inserted into patients, the deposition of macromolecules such as fibrinogen and immunoglobin on their surfaces makes it easier for them to be prone to bacteria colonization leading to the formation of biofilms. The formation of biofilms on medical devices has led to a series of device-related infections which are usually difficult to eradicate and sometimes cause the death of patients. These infections require surgical replacements along with prolonged antibiotic therapy, which would incur additional health costs. It is, therefore, necessary to prevent device-related infections by inhibiting the formation of biofilms using intelligent technology. Antibiotic resistance of bacteria is also a major threat due to overuse. Different antimicrobial agents have been applied to microbial infections. They include conventional antibiotics like rifampicin. The use of conventional antibiotics like rifampicin has raised concerns as some have been found to have hepatic and nephrotoxic effects due to overuse. Hence, there is also a need for proper delivery of these antibiotics. Different techniques have been developed to encapsulate and slowly release antimicrobial agents, thus reducing host cytotoxicity. Examples of delivery systems are solid lipid nanoparticles, hydrogels, micelles, and polymeric nanoparticles. The different ways by which drugs are released from polymeric nanoparticles include diffusion-based release, elution-based release, and chemical/stimuli-responsive release. Polymeric nanoparticles have gained a lot of research interest as they are basically made from biodegradable polymers. An example of such a biodegradable polymer is polycaprolactone (PCL). PCL degrades slowly by hydrolysis but is often sensitive and responsive to stimuli like enzymes to release encapsulants for antimicrobial therapy. This study presents the synthesis of PCL nanoparticles loaded with rifampicin and the on-demand release of rifampicin for treating staphylococcus aureus infections.

Keywords: enzyme, Staphylococcus aureus, PCL, rifampicin

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Authors: Mario Pérez-Won, Anais Palma-Acevedo, Luis González-Cavieres, Roberto Lemus-Mondaca, Gipsy Tabilo-Munizaga

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The Chilean abalone (Concholepas Concholepas) is a gastropod mollusk; it has a high commercial value due to the qualities of its meat, especially hardness, as a critical acceptance parameter. However, its main problem is its short shelf-life which is usually extended using traditional technologies with high energy consumption. Therefore, applying different technologies for the pre-treatment and drying process is necessary. In this research, pulsed electric field (PEF) was used as a pre-treatment for vacuum microwave drying (VMD), freeze-drying (FD), and hot-air drying (HAD). Drying conditions and characteristics were set according to previous experiments. The Drying samples were analyzed in terms of physical quality (color, texture, microstructure, and rehydration capacity), protein quality (degree of hydrolysis and computer protein efficiency ratio), and energy parameters. Regarding quality, the treatment that obtained lower harness was PEF+FD (195 N ± 10), the lowest change of color was for treatment PEF+VMD (ΔE: 17 ± 1.5), and the best rehydration capacity was for treatment PEF+VMD (1.2 h for equilibrium). For protein quality, the highest Computer-Protein Efficiency Ratio was the sample 2.0 kV/ cm of PEF (index of 4.18 ± 0.26 at the end of the digestion). Moreover, about energetic consumption, results show that VMD decreases the drying process by 97% whether PEF was used or not. Consequently, it is possible to conclude that using PEF as a pre-treatment for VMD and FD treatments has advantages that must be used following the consumer’s needs or preferences.

Keywords: chilean abalone, freeze-drying, proteins, pulsed electric fields

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Authors: Naureen Akhtar

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The stability of the protein in detergent-containing solution is the key for its successful crystallisation. Fluorescence-detection size-exclusion chromatography (FSEC) is a potential approach for screening monodispersity as well as the stability of protein in a detergent-containing-solution. In this present study, covalently linked Green Fluorescent Protein (GFP) to bacterial nitrate transporter, NarU from Escherichia coli was studied for pre-crystallisation trials by FSEC. Immobilised metal ion affinity chromatography (IMAC) and gel filtration were employed for their purification. The main objectives of this study were over-expression, detergent screening and crystallisation of nitrate transporter proteins. This study could not produce enough proteins that could realistically be taken forward to achieve the objectives set for this particular research. In future work, different combinations of variables like vectors, tags, creation of mutant proteins, host cells, position of GFP (N- or C-terminal) and/or membrane proteins would be tried to determine the best combination as the principle of technique is still promising.

Keywords: transporters, detergents, over-expression, crystallography

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Authors: M. Kowalski, M. Brodowski, K. Dziabowska, E. Czaczyk, W. Bialobrzeska, N. Malinowska, S. Zoledowska, R. Bogdanowicz, D. Nidzworski

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Boron-doped-carbon nanowall (BCNW) electrodes are recently in much interest among scientists. BCNWs are good candidates for biosensor purposes as they possess interesting electrochemical characteristics like a wide potential range and the low difference between redox peaks. Moreover, from technical parameters, they are mechanically resistant and very tough. The production process of the microwave plasma-enhanced chemical vapor deposition (MPECVD) allows boron to build into the structure of the diamond being formed. The effect is the formation of flat, long structures with sharp ends. The potential of these electrodes was checked in the biosensing field. The procedure of simple carbon electrodes modification by antibodies was adopted to BCNW for specific antigen recognition. Surface protein D deriving from H. influenzae pathogenic bacteria was chosen as a target analyte. The electrode was first modified with the aminobenzoic acid diazonium salt by electrografting (electrochemical reduction), next anti-protein D antibodies were linked via 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride/N-hydroxysuccinimide (EDC/NHS) chemistry, and free sites were blocked by BSA. Cyclic voltammetry measurements confirmed the proper electrode modification. Electrochemical impedance spectroscopy records indicated protein detection. The sensor was proven to detect protein D in femtograms. This work was supported by the National Centre for Research and Development (NCBR) TECHMATSTRATEG 1/347324/12/NCBR/ 2017.

Keywords: anti-protein D antibodies, boron-doped carbon nanowall, impedance spectroscopy, Haemophilus influenzae.

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Authors: Krasimir Ivanov, Elitsa Kolentsova, Nguyen Nguyen, Alexander Peltekov, Violina Angelova

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The nanosized zinc hydroxide nitrate has been recently estimated as perspective foliar fertilizer, which has improved zinc solubility, but low phytotoxicity, in comparison with ZnO and other Zn containing compounds. The main problem is obtaining of stable particles with dimensions less than 100 nm. This work studies the effect of preparation conditions on the chemical compositions and particle size of the zinc hydroxide nitrates, prepared by precipitation. Zn(NO3)2.6H2O and NaOH with concentrations, ranged from 0.2 to 3.2M and the initial OH/Zn ratio from 0.5 to 1.6 were used at temperatures from 20 to 60 °C. All samples were characterized in detail by X-ray diffraction, scanning electron microscopy, differential thermal analysis and ICP. Stability and distribution of the zinc hydroxide nitrate particles were estimated too.

Keywords: zinc hydroxide nitrate, nanoparticles, preparation, foliar fertilizer

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Authors: Elmira Solati, Atousa Mehrani, Davoud Dorranian

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Generation of ZnO-Au nanocomposite under laser irradiation of a mixture of the ZnO and Au colloidal suspensions are experimentally investigated. In this work, firstly ZnO and Au nanoparticles are prepared by pulsed laser ablation of the corresponding metals in water using the 1064 nm wavelength of Nd:YAG laser. In a second step, the produced ZnO and Au colloidal suspensions were mixed in different volumetric ratio and irradiated using the second harmonic of a Nd:YAG laser operating at 532 nm wavelength. The changes in the size of the nanostructure and optical properties of the ZnO-Au nanocomposite are studied as a function of the volumetric ratio of ZnO and Au colloidal suspensions. The crystalline structure of the ZnO-Au nanocomposites was analyzed by X-ray diffraction (XRD). The optical properties of the samples were examined at room temperature by a UV-Vis-NIR absorption spectrophotometer. Transmission electron microscopy (TEM) was done by placing a drop of the concentrated suspension on a carbon-coated copper grid. To further confirm the morphology of ZnO-Au nanocomposites, we performed Scanning electron microscopy (SEM) analysis. Room temperature photoluminescence (PL) of the ZnO-Au nanocomposites was measured to characterize the luminescence properties of the ZnO-Au nanocomposites. The ZnO-Au nanocomposites were characterized by Fourier transform infrared (FTIR) spectroscopy. The X-ray diffraction pattern shows that the ZnO-Au nanocomposites had the polycrystalline structure of Au. The behavior observed by images of transmission electron microscope reveals that soldering of Au and ZnO nanoparticles include their adhesion. The plasmon peak in ZnO-Au nanocomposites was red-shifted and broadened in comparison with pure Au nanoparticles. By using the Tauc’s equation, the band gap energy for ZnO-Au nanocomposites is calculated to be 3.15–3.27 eV. In this work, the formation of ZnO-Au nanocomposites shifts the FTIR peak of metal oxide bands to higher wavenumbers. PL spectra of the ZnO-Au nanocomposites show that several weak peaks in the ultraviolet region and several relatively strong peaks in the visible region. SEM image indicates that the morphology of ZnO-Au nanocomposites produced in water was spherical. The TEM images of ZnO-Au nanocomposites demonstrate that with increasing the volumetric ratio of Au colloidal suspension the adhesion increased. According to the size distribution graphs of ZnO-Au nanocomposites with increasing the volumetric ratio of Au colloidal suspension the amount of ZnO-Au nanocomposites with the smaller size is further.

Keywords: Au nanoparticles, pulsed laser ablation, ZnO-Au nanocomposites, ZnO nanoparticles

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Authors: Shahram Naghizadeh Raeisi, Ali Alghooneh

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The loss of curd cohesiveness and syneresis are two common problems in the ultrafiltered cheese industry. In this study, by using membrane technology and protein modification, a modified cheese was developed and its properties were compared with a control sample. In order to decrease the lactose content and adjust the protein, acidity, dry matter and milk minerals, a combination of ultrafiltration, nanofiltration and reverse osmosis technologies was employed. For protein modification, a two-stage chemical and enzymatic reaction was employed before and after ultrafiltration. The physicochemical and microstructural properties of the modified ultrafiltered cheese were compared with the control one. Results showed that the modified protein enhanced the functional properties of the final cheese significantly (pvalue< 0.05), even if the protein content was 50% lower than the control one. The modified cheese showed 21 ± 0.70, 18 ± 1.10 & 25±1.65% higher hardness, cohesiveness and water-holding capacity values, respectively, than the control sample. This behavior could be explained by the developed microstructure of the gel network. Furthermore, chemical-enzymatic modification of milk protein induced a significant change in the network parameter of the final cheese. In this way, the indices of network linkage strength, network linkage density, and time scale of junctions were 10.34 ± 0.52, 68.50 ± 2.10 & 82.21 ± 3.85% higher than the control sample, whereas the distance between adjacent linkages was 16.77 ± 1.10% lower than the control sample. These results were supported by the results of the textural analysis. A non-linear viscoelastic study showed a triangle waveform stress of the modified protein contained cheese, while the control sample showed rectangular waveform stress, which suggested a better sliceability of the modified cheese. Moreover, to study the shelf life of the products, the acidity, as well as molds and yeast population, were determined in 120 days. It’s worth mentioning that the lactose content of modified cheese was adjusted at 2.5% before fermentation, while the lactose of the control one was at 4.5%. The control sample showed 8 weeks shelf life, while the shelf life of the modified cheese was 18 weeks in the refrigerator. During 18 weeks, the acidity of modified and control samples increased from 82 ± 1.50 to 94 ± 2.20 °D and 88 ± 1.64 to 194 ± 5.10 °D, respectively. The mold and yeast populations, with time, followed the semicircular shape model (R2 = 0.92, R2adj = 0.89, RMSE = 1.25). Furthermore, the mold and yeast counts and their growth rate in the modified cheese were lower than those for control one; Aforementioned result could be explained by the shortage of the source of energy for the microorganism in the modified cheese. The lactose content of the modified sample was less than 0.2 ± 0.05% at the end of fermentation, while this was 3.7 ± 0.68% in the control sample.

Keywords: non-linear viscoelastic, protein modification, semicircular shape model, ultrafiltered cheese

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Authors: Nedal N. Marei, Nashaat Nassar

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Many treatment techniques have been used to remove the soluble pollutants from wastewater as; dyes and metal ions which could be found in rich amount in the used water of the textile and tanneries industry. The effluents from these industries are complex, containing a wide variety of dyes and other contaminants, such as dispersants, acids, bases, salts, detergents, humectants, oxidants, and others. These techniques can be divided into physical, chemical, and biological methods. Adsorption has been developed as an efficient method for the removal of heavy metals from contaminated water and soil. It is now recognized as an effective method for the removal of both organic and inorganic pollutants from wastewaters. Nanosize materials are new functional materials, which offer high surface area and have come up as effective adsorbents. Nano alumina is one of the most important ceramic materials widely used as an electrical insulator, presenting exceptionally high resistance to chemical agents, as well as giving excellent performance as a catalyst for many chemical reactions, in microelectronic, membrane applications, and water and wastewater treatment. In this study, methylene blue (MB) dye has been used as model dye of textile wastewater in order to synthesize a synthetic MB wastewater. NiO nanoparticles were added in small percentage in the sand packed bed adsorption columns to remove the MB from the synthetic textile wastewater. Moreover, different parameters have been evaluated; flow of the synthetic wastewater, pH, height of the bed, percentage of the NiO to the sand in the packed material. Different mathematical models where employed to find the proper model which describe the experimental data and help to analyze the mechanism of the MB adsorption. This study will provide good understanding of the dyes adsorption using metal oxide nanoparticles in the classical sand bed.

Keywords: adsorption, column, nanoparticles, methylene

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Authors: Zi-Han Li, Lu Fang, Liang Wu, Dong-Yuan Chang, Manyuan Dong, Liang Ji, Qi Zhang, Ming-Hui Zhao, Sydney C. W. Tang, Lemin Zheng, Min Chen

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Uncoupling of mitochondrial respiration by chemical uncouplers has proven effective in ameliorating obesity, insulin resistance, and hyperglycemia, which were risk factors for diabetic nephropathy (DN). Recently, we found that ANXA1 could improve mitochondrial function to mitigate DN progression. However, the underlying mechanism is not fully clear yet. Here, we identified uncoupling protein 1 (UCP1), an inner membrane protein of mitochondria, as a key to mitochondrial homeostasis improved by ANXA1. Specifically, ANXA1 attenuated mitochondrial dysfunction via appropriately upregulating UCP1 by stabilizing its transcription factor GATA binding protein 3 (GATA3) by combining it with thioredoxin. Moreover, specific overexpression of UCP1 in the renal cortex rescued renal injuries in diabetic Anxa1-KO mice. UCP1 deletion aggravated renal injuries in HFD/STZ-induced diabetic mice. Mechanistically, UCP1 reduced mitochondrial fission through the aristaless-related homeobox (ARX)/cardiolipin synthase 1 (CRLS1) pathway. Therapeutically, CL316243, a UCP1 agonist, could attenuate established DN in db/db mice. This work established an alternative principle to harness the power of uncouplers for the treatment of DN.

Keywords: diabetic nephropathy, uncoupling protein 1, mitochondrial homeostasis, cardiolipin metabolism

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Authors: Sadegh Lotfieblisofla, Arash Khodabakhshi

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Tissue plasminogen activator (tPA) as a serine protease plays an important role in the fibrinolytic system and the dissolution of fibrin clots in human body. The production of this drug in plants such as tobacco could reduce its production costs. In this study, expression of tPA gene and protein targeting to different plant cell compartments, using various signal peptides has been investigated. For high level of expression, Kozak sequence was used after CaMV35S in the beginning of the gene. In order to design the final construction, Extensin, KDEL (amino acid sequence including Lys-Asp-Glu-Leu) and SP (γ-zein signal peptide coding sequence) were used as leader signals to conduct this protein into apoplast, endoplasmic reticulum and cytosol spaces, respectively. Cloned human tPA gene under the CaMV (Cauliflower mosaic virus) 35S promoter and NOS (Nopaline Synthase) terminator into pBI121 plasmid was transferred into tobacco explants by Agrobacterium tumefaciens strain LBA₄₄₀₄. The presence and copy number of genes in transgenic tobacco was proved by Southern blotting. Enzymatic activity of the rt-PA protein in transgenic plants compared to non-transgenic plants was confirmed by Zymography assay. The presence and amount of rt-PA recombinant protein in plants was estimated by ELISA analysis on crude protein extract of transgenic tobacco using a specific antibody. The yield of recombinant tPA in transgenic tobacco for SP, KDEL, Extensin signals were counted 0.50, 0.68, 0.69 microgram per milligram of total soluble proteins.

Keywords: tPA, recombinant, transgenic, tobacco

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Authors: Shivani A. Singh, Pravin S. More

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In the present inspection, we indicate the falsification of Fe-doped graphene nanoparticles by modified Hummers method. Structural and physiochemical properties of the resulting pallets were explored with the help of ultraviolet-visible spectroscopy (UV-Vis), Fourier transform infrared spectroscopy (FTIR), X-ray powder diffraction (XRD) and scanning electron microscopy (SEM), Photoluminescence spectroscopy (PL) for graphene sample exhibits absorption peaks ~248nm. Pure graphene shows PL peak at 348 nm. After doping of Fe with graphene the PL peak shifted from 348 nm to 332 nm. The oxidation degree, i.e. the relative amount of oxygen functional groups was estimated from the relative intensities of the oxygen related bands (ORB) in the FTIR measurements. These analyses show that this modified material can be useful for gas sensing applications and to be used in diverse areas.

Keywords: chemical doping, graphene, gas sensing, sensing

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Authors: Mohamad Ammar Ayass, Natalya Griko, Victor Pashkov, Wanying Cao, Kevin Zhu, Jin Zhang, Lina Abi Mosleh

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Respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent for coronavirus disease 2019 (COVID-19). Early evidence pointed at the angiotensin-converting enzyme 2 (ACE-2) expressed on the epithelial cells of the lung as the main entry point of SARS-CoV-2 into the cells. The viral entry is mediated by the binding of the Receptor Binding Domain (RBD) of the spike protein that is expressed on the surface of the virus to the ACE-2 receptor. As the number of SARS-CoV-2 variants continues to increase, mutations arising in the RBD of SARS-CoV-2 may lead to the ineffectiveness of RBD targeted neutralizing antibodies. To address this limitation, the objective of this study is to develop a combination of aptamers that target different regions of the RBD, preventing the binding of the spike protein to ACE-2 receptor and subsequent viral entry and replication. A safe and innovative biomedical tool was developed to inhibit viral infection and reduce the harms of COVID-19. In the present study, DNA aptamers were developed against a recombinant trimer S protein using the Systematic Evolution of Ligands by Exponential enrichment (SELEX). Negative selection was introduced at round number 7 to select for aptamers that bind specifically to the RBD domain. A series of 9 aptamers (ADI2010, ADI2011, ADI201L, ADI203L, ADI205L, ADIR68, ADIR74, ADIR80, ADIR83) were selected and characterized with high binding affinity and specificity to the RBD of the spike protein. Aptamers (ADI25, ADI2009, ADI203L) were able to bind and pull down endogenous spike protein expressed on the surface of SARS-CoV-2 virus in COVID-19 positive patient samples and determined by liquid chromatography- tandem mass spectrometry analysis (LC-MS/MS). LC-MS/MS data confirmed that aptamers can bind to the RBD of the spike protein. Furthermore, results indicated that the combination of the 9 best aptamers inhibited the binding of the purified trimer spike protein to the ACE-2 receptor found on the surface of Vero E6 cells. In the same experiment, the combined aptamers displayed a better neutralizing effect than antibodies. The data suggests that the selected aptamers could be used in therapy to neutralize the effect of the SARS-CoV-2 virus by inhibiting the interaction between the RBD and ACE-2 receptor, preventing viral entry into target cells and therefore blocking viral replication.

Keywords: aptamer, ACE-2 receptor, binding inhibitor, COVID-19, spike protein, SARS-CoV-2, treatment

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Authors: Hanan. Al Chaghouri, Mohammad Azad Malik, P. John Thomas, Paul O’Brien

Abstract:

The process of assembling metal nanoparticles at the interface of two liquids has received a great interest over the past few years due to a wide range of important applications and their unusual properties compared to bulk materials. We present a low cost, simple and cheap synthesis of metal nanoparticles, core/shell structures and semiconductors followed by assembly of these particles between immiscible liquids. The aim of this talk is divided to three parts: firstly, to describe the achievement of a closed loop recycling for producing cadmium sulphide as powders and/or nanostructured thin films for solar cells or other optoelectronic devices applications by using a different chain length of commercially available secondary amines of dithiocarbamato complexes. The approach can be extended to other metal sulphides such as those of Zn, Pb, Cu, or Fe and many transition metals and oxides. Secondly, to synthesis significantly cheaper magnetic particles suited for the mass market. Ni/NiO nanoparticles with ferromagnetic properties at room temperature were among the smallest and strongest magnets (5 nm) were made in solution. The applications of this work can be applied to produce viable storage devices and the other possibility is to disperse these nanocrystals in solution and use it to make ferro-fluids which have a number of mature applications. The third part is about preparing and assembling of submicron silver, cobalt and nickel particles by using polyol methods and liquid/liquid interface, respectively. Noble metal like gold, copper and silver are suitable for plasmonic thin film solar cells because of their low resistivity and strong interactions with visible light waves. Silver is the best choice for solar cell application since it has low absorption losses and high radiative efficiency compared to gold and copper. Assembled cobalt and nickel as films are promising for spintronic, magnetic and magneto-electronic and biomedics.

Keywords: assembling nanoparticles, liquid/liquid interface, thin film, core/shell, solar cells, recording media

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Authors: Bekan Bogale, Tsegaye Girma Asere, Tilahun Yai, Fekadu Melak

Abstract:

Aims: To study photocatalytic degradation of methylene blue dye on cuprous oxide/graphene nanocomposite. Background: Cuprous oxide (Cu2O) nanoparticles are among the metal oxides that demonstrated photocatalytic activity. However, the stability of Cu2O nanoparticles due to the fast recombination rate of electron/hole pairs remains a significant challenge in their photocatalytic applications. This, in turn, leads to mismatching of the effective bandgap separation, tending to reduce the photocatalytic activity of the desired organic waste (MB). To overcome these limitations, graphene has been combined with cuprous oxides, resulting in cuprous oxide/graphene nanocomposite as a promising photocatalyst. Objective: In this study, Cu2O/graphene nanocomposite was synthesized and evaluated for its photocatalytic performance of methylene blue (MB) dye degradation. Method: Cu2O/graphene nanocomposites were synthesized from graphite powder and copper nitrate using the facile sol-gel method. Batch experiments have been conducted to assess the applications of the nanocomposites for MB degradation. Parameters such as contact time, catalyst dosage, and pH of the solution were optimized for maximum MB degradation. The prepared nanocomposites were characterized by using UV-Vis, FTIR, XRD, and SEM. The photocatalytic performance of Cu2O/graphene nanocomposites was compared against Cu2O nanoparticles for cationic MB dye degradation. Results: Cu2O/graphene nanocomposite exhibits higher photocatalytic activity for MB degradation (with a degradation efficiency of 94%) than pure Cu2O nanoparticles (67%). This has been accomplished after 180 min of irradiation under visible light. The kinetics of MB degradation by Cu2O/graphene composites can be demonstrated by the second-order kinetic model. The synthesized nanocomposite can be used for more than three cycles of photocatalytic MB degradation. Conclusion: This work indicated new insights into Cu2O/graphene nanocomposite as high-performance in photocatalysis to degrade MB, playing a great role in environmental protection in relation to MB dye.

Keywords: methylene blue, photocatalysis, cuprous oxide, graphene nanocomposite

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Authors: Reynaldo Esquivel, Pedro Hernandez, Aaron Martinez-Higareda, Sergio Tena-Cano, Enrique Alvarez-Ramos, Armando Lucero-Acuna

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Stimuli-responsive nanomaterials play an essential role in loading, transporting and well-distribution of anti-cancer compounds in the cellular surroundings. The outstanding properties as the Lower Critical Solution Temperature (LCST), hydrolytic cleavage and protonation/deprotonation cycle, govern the release and delivery mechanisms of payloads. In this contribution, we experimentally determine the load efficiency and release of antineoplastic Vincristine Sulfate into PNIPAM-Interpenetrated-Chitosan (PIntC) nanoparticles. Structural analysis was performed by Fourier Transform Infrared Spectroscopy (FT-IR) and Proton Nuclear Magnetic Resonance (1HNMR). ζ-Potential (ζ) and Hydrodynamic diameter (DH) measurements were monitored by Electrophoretic Mobility (EM) and Dynamic Light scattering (DLS) respectively. Mathematical analysis of the release pharmacokinetics reveals a three-phase model above LCST, while a monophasic of Vincristine release model was observed at 32 °C. Cytotoxic essays reveal a noticeable enhancement of Vincristine effectiveness at low drug concentration on HeLa cervix cancer and MDA-MB-231 breast cancer.

Keywords: nanoparticles, vincristine, drug delivery, PNIPAM

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Authors: Ç. Gökçek-Saraç, Ş. Özen, N. Derin

Abstract:

The N-methyl-D-aspartate (NMDA)-dependent pathway is the major intracellular signaling pathway implemented in both short- and long-term memory formation in the hippocampus which is the most studied brain structure because of its well documented role in learning and memory. However, little is known about the effects of RF-EMR exposure on NMDA receptor signaling pathway including activation of protein kinases, notably Ca²⁺/calmodulin-dependent protein kinase II alpha (CaMKIIα). The aim of the present study was to investigate the effects of acute and chronic 900 MHz RF-EMR exposure on both passive avoidance behaviour and hippocampal levels of CaMKIIα and its phosphorylated form (pCaMKIIα). Rats were divided into the following groups: Sham rats, and rats exposed to 900 MHz RF-EMR for 2 h/day for 1 week (acute group) or 10 weeks (chronic group), respectively. Passive avoidance task was used as a behavioural method. The hippocampal levels of selected kinases were measured using Western Blotting technique. The results of passive avoidance task showed that both acute and chronic exposure to 900 MHz RF-EMR can impair passive avoidance behaviour with minor effects on chronic group of rats. The analysis of western blot data of selected protein kinases demonstrated that hippocampal levels of CaMKIIα and pCaMKIIα were significantly higher in chronic group of rats as compared to acute groups. Taken together, these findings demonstrated that different duration times (1 week vs 10 weeks) of 900 MHz RF-EMR exposure have different effects on both passive avoidance behaviour of rats and hippocampal levels of selected protein kinases.

Keywords: hippocampus, protein kinase, rat, RF-EMR

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Authors: P. F. Msomi, P. T. Nonjola, P. G. Ndungu, J. Ramontja

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In view of the projected global energy demand and increasing levels of greenhouse gases and pollutants issues have inspired an intense search for alternative new energy technologies, which will provide clean, low cost and environmentally friendly solutions to meet the end user requirements. Alkaline anion exchange membrane fuel cells (AAEMFC) have been recognized as ideal candidates for the generation of such clean energy for future stationary and mobile applications due to their many advantages. The key component of the AAEMFC is the anion exchange membrane (AEM). In this report, a series of quaternized poly (2.6 dimethyl – 1.4 phenylene oxide)/ polysulfone (QPPO/PSF) blend anionic exchange membranes (AEM) were successfully fabricated and characterized for alkaline fuel cell application. Zinc Oxide (ZnO) nanoparticles were introduced in the polymer matrix to enhance the intrinsic properties of the AEM. The characteristic properties of the QPPO/PSF and QPPO/PSF-ZnO blend membrane were investigated with X-ray diffraction (XRD), thermogravimetric analysis (TGA) scanning electron microscope (SEM) and contact angle (CA). To confirm successful quaternisation, FT-IR spectroscopy and proton nuclear magnetic resonance (1H NMR) were used. Other properties such as ion exchange capacity (IEC), water uptake, contact angle and ion conductivity (IC) were also undertaken to check if the prepared nanocomposite materials are suitable for fuel cell application. The membrane intrinsic properties were found to be enhanced by the addition of ZnO nanoparticles. The addition of ZnO nanoparticles resulted to a highest IEC of 3.72 mmol/g and a 30-fold IC increase of the nanocomposite due to its lower methanol permeability. The above results indicate that QPPO/PSF-ZnO is a good candidate for AAEMFC application.

Keywords: anion exchange membrane, fuel cell, zinc oxide nanoparticle, nanocomposite

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Authors: F. Z. Benabid, S. Kridi, F. Zouai, D. Benachour

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The aim of present work is to characterize the PVDF/TiO2 blends as nanocomposites, and study the effect of TiO2 on properties of different compositions and the evaluation of the effectiveness of the method used for filler treatment. Nanocomposite samples were synthesized by molten route in an internal mixer. The TiO2 nanoparticles were treated with stearic acid in order to obtain a good dispersion, and the demonstration of the effectiveness of the treatment on the morphology and roughness of the nanofiller was established by microstructural analysis by FTIR and AFM. The various developed nanocomposite compositions were characterized by different methods; i.e. FTIR, XRD, SEM and optical microscopy. Rheological, dielectric and mechanical studies were also performed. The results showed a remarkable increase in the crystallinity of the PVDF/neat TiO2 nanocomposite containing 1 wt% loading of filler, due to the nucleation effect of TiO2 nanoparticles. A good dispersion was obtained in PVDF/treated TiO2 nanocomposites. The rheological study showed an increase in the fluidity in all developed nanocomposite compositions, involved by the orientation of TiO2 nanoparticles in the flow direction. The dielectric study revealed an increase in electrical conductivity in PVDF/neat TiO2 nanocomposites. However, in PVDF/ treated TiO2 nanocomposites, the electrical conductivity was decreased by the addition of 0.5 and 2 wt% loading of filler.

Keywords: nanocomposites, PVDF, TiO2, comixing, mechanical treatment

Procedia PDF Downloads 285

Authors: Salahaedin Waiezi, Nik Ahmad Nizam Nik Malek, Hassan Abdelmagid Elzamzami, Shahrulnizahana Mohammad Din

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A green and safe approach to the synthesis of silver nanoparticles (AgNP) can be performed using plant leaf extract as the reducing agent. Hence, this paper reports the biosynthesis of AgNP using Clinacanthus nutans plant extract. C. nutans is known as belalai gajah in Malaysia and is widely used as a medicinal herb locally. The biosynthesized AgNP, using C. nutans aqueous extract at pH 10, with the reaction temperature of 70°C and 48 h reaction time, was characterized by UV-Vis spectroscopy, X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), energy dispersive X-ray (EDX), and transmission electron microscope (TEM). A peak appeared in the UV-Vis spectra at around 400 nm, while XRD confirmed the crystal structure of AgNP, with the average size between 20 to 30 nm, as shown in FESEM and TEM. The antibacterial activity of the biosynthesized AgNP, which was performed using the disc diffusion technique (DDT) indicated effective inhibition against Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. In contrast, minimal antibacterial activity was detected against Enterococcus faecalis and methicillin-resistant Staphylococcus aureus (MRSA). In general, AgNP produced using C. nutans leaf extract possesses potential antibacterial activity.

Keywords: silver nanoparticles, Clinacanthus nutans, antibacterial agent, biosynthesis

Procedia PDF Downloads 182

Authors: Nasser A. Al-Shabib

Abstract:

Food allergens are most common non-native form when exposed to the immune system. Most food proteins undergo various treatments (e.g. thermal or proteolytic processing) during food manufacturing. Such treatments have the potential to impact the chemical structure of food allergens so as to convert them to more denatured or unfolded forms. The conformational changes in the proteins may affect the allergenicity of treated-allergens. However, most allergenic proteins possess high resistance against thermal modification or digestive enzymes. In the present study, ovomucoid (a major allergenic protein of egg white) was heated in phosphate-buffered saline (pH 7.4) at different temperatures, aqueous solutions and on different surfaces for various times. The results indicated that different antibody-based methods had different sensitivities in detecting the heated ovomucoid. When using one particular immunoassay‚ the immunoreactivity of ovomucoid increased rapidly after heating in water whereas immunoreactivity declined after heating in alkaline buffer (pH 10). Ovomucoid appeared more immunoreactive when dissolved in PBS (pH 7.4) and heated on a stainless steel surface. To the best of our knowledge‚ this is the first time that antibody-based methods have been applied for the detection of ovomucoid adsorbed onto different surfaces under various conditions. The results obtained suggest that use of antibodies to detect ovomucoid after food processing may be problematic. False assurance will be given with the use of inappropriate‚ non-validated immunoassays such as those available commercially as ‘Swab’ tests. A greater understanding of antibody-protein interaction after processing of a protein is required.

Keywords: ovomucoid, thermal treatment, solutions, surfaces

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Authors: Han-Bin Kim, Sooim Shin, Moonsung Choi

Abstract:

There is a growing interest in introducing a desired functional group on enzymes in the field of protein engineering. In here, various redox centers were newly created using histidine tag, which is widely used for protein purification, plus cobalt in one of cupredoxins, amicyanin. The coordination of Cobalt-His tag and reactivity of the Co²⁺ loaded His-tag also were characterized. 3xHis-tag, 6xHis-tag, and 9xHis-tag were introduced on amicyanin by site-directed mutagenesis, and then Co²⁺ was loaded on each His-tagged amicyanin. The spectral changes at 330 nm corresponding to cobalt binding on His-tag site indicated the binding ratio of 3xHis-tag, 6xHis-tag, and 9xHis-tag to cobalt as 1:1, 1:2, 1:3 respectively. Based on kinetic studies of binding cobalt to 3xHis-tag, 6xHis-tag, and 9xHis-tagged amicyanin, the nature of the sites was elucidated. In addition, internal electron transfer properties between Cu¹⁺ site and engineered site of amicyanin were determined. These results provide insight into improvement of metal coordination and alternation of the redox properties of metal as a new catalytic site on proteins.

Keywords: amicyanin, cobalt, histidine, protein engineering

Procedia PDF Downloads 145