Functional characterization of a manganese superoxide dismutase from Avicennia marina: insights into its role in salt, hydrogen peroxide, and heavy metal tolerance.

Avicennia marina is a salt-tolerance plant with high antioxidant and antibacterial potential. In the present work, a gene encoding MnSOD from Avicennia marina (AmSOD2) was cloned in the expression vectors pET28a. The resulting constructs were transformed into Escherichia coli strains Rosetta (DE3). Following the induction with Isopropyl ß-D-1-thiogalactopyranoside, the protein His-AmSOD2 was expressed but dominantly found in the insoluble fraction of strain R-AmSOD2. Due to detection of mitochondrial transit peptide in the amino acid sequence of AmSOD2, the transit peptide was removed and AmSOD2 without transit peptide (tAmSOD2) was expressed in E. coli and dominantly found in the soluble fraction. The enzyme His-tAmSOD2 exhibited a molecular mass of 116 kDa in native condition. Nevertheless, in reducing conditions the molecular mass is 28 kDa indicating the enzyme His-tAmSOD2 is a tetramer protein. As shown by ICP analysis there is one mole Mn2+ in each monomer. The Pure His-tAmSOD2 was highly active in vitro, however the activity was almost three-fold lower than His-AmSOD1. Whereas the high stability of the recombinant His-AmSOD1was previously shown after incubation in a broad range pH and high temperature, His-tAmSOD2 was stable up to 50 °C and pH 6 for 1 h. The gene expression analysis showed that the gene encoding AmSOD2 is expressed in root, shoot and leaves of A. marina. In addition, the results show that the expression in the leaves was enhanced after treatment of plant with NaCl, H2O2, Cd2+ and Ni2+ indicating the important role of MnSOD in the resistant mechanism of mangroves.

Introduction of nitrogen doped graphene nanosheets as efficient adsorbents for nitrate removal from aqueous samples.

PURPOSE: Introducing and developing new kinds of adsorbents are always a significant challenge in water treatments. In this work, for the first time, graphene oxide (GO), nitrogen-doped graphene oxide (ND-GO), highly nitrogen-doped graphene oxide (HND-GO), and 3D high nitrogen-doped graphene oxide (3D-HND-GO) were synthesized and comparatively evaluated in the removal of nitrate content of tap and underground waters. METHODS: The removal of the target analyte was performed through a batch adsorption approach, and the factors influencing its removal efficiency (i.e., initial pH of the sample, primary concentrations of nitrate, amount of adsorbent, and contact time) were evaluated through a central composite design (CCD) and response surface methodology (RSM). RESULTS: Based on the results, 3D-HND-GO showed the highest removal efficiency in comparison with the other mentioned nanoparticles. The nitrate removal using this adsorbent was modeled successfully so that R 2, adjusted R 2, and predicted R 2 values were 0.9717, 0.9508, and 0.9010, respectively. In addition, the optimal removal condition was achieved using the Nelder-Mead non-linear optimization algorithm as follow: the initial concentrations of nitrate (expressed as nitrogen): 15.0 mg/mL, the amount of the adsorbent: 2.0 mg/mL; pH of the sample: 3.0; and the contact time: 20.0 min. Under this optimal condition, the actual removal result (92.5 ± 4.0%) was in good agreement with the expected value (94.8 ± 5.1%). Additional studies were also performed to comprehensibly evaluate the adsorption activity of the adsorbent (e.g., kinetic, isotherm, and desorption parameters). The adsorption isotherm complied with the Langmuir model illustrating the considerable mono-layer adsorption capacities for the target ions with qm of 8.7 mg/g. The adsorption process was indicated to obey a pseudo 2nd order kinetic model, with the rate-limiting step for the adsorption phase. CONCLUSIONS: This study revealed which 3D-HND-G leads to improved yield in the nitrate ions elimination, particularly at acidic media, which was related to the enhanced dispersibility and larger surface area. The adsorbent was further successfully used for treating tap and underground water samples. At the present moment, research as grown to modify 3D-HND-G in orders to increase the potentiality for industrial applications. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s40201-021-00741-7.

Polylactic Acid Piezo-Biopolymers: Chemistry, Structural Evolution, Fabrication Methods, and Tissue Engineering Applications.

Polylactide acid (PLA), as an FDA-approved biomaterial, has been widely applied due to its unique merits, such as its biocompatibility, biodegradability, and piezoelectricity. Numerous utilizations, including sensors, actuators, and bio-application-its most exciting application to promote cell migration, differentiation, growth, and protein-surface interaction-originate from the piezoelectricity effect. Since PLA exhibits piezoelectricity in both crystalline structure and an amorphous state, it is crucial to study it closely to understand the source of such a phenomenon. In this respect, in the current study, we first reviewed the methods promoting piezoelectricity. The present work is a comprehensive review that was conducted to promote the low piezoelectric constant of PLA in numerous procedures. In this respect, its chemistry and structural origins have been explored in detail. Combining any other variables to induce a specific application or to improve any PLA barriers, namely, its hydrophobicity, poor electrical conductivity, or the tuning of its mechanical properties, especially in the application of cardiovascular tissue engineering, is also discussed wherever relevant.

Solid-phase microextraction of methadone by using a chitosan nanocomposite incorporated with Polyoxomolibdate nanocluster/Graphene oxide.

In the present study, we report on the simple sol-gel preparation of a nanocomposite composed of chitosan/ polyoxometalate /graphene oxide, and its application in the headspace solid-phase microextraction combined with the ion mobility spectrometry for the analysis of methadone in biological matrices. The developed nanocomposite was characterized through the infrared spectroscopy and thermogravimetric analyses. The ternary nanocomposite coating offers good mechanical and thermal stability and high extraction efficiency thanks to its large specific surface. A central composite statistical design was used to study the main variables affecting the extraction efficiency. Afterward, to study the relationship between different input and output variables as well as to identify the optimal operating conditions, response surface methodology was used, whereby a second-order polynomial equation was fit to the experimental data. The optimized extraction conditions were as follows: temperature, 70°C; extraction time, 15 min; and concentration of NaCl, 5%w/v. The detection limit of 0.12 ng/mL was obtained at the optimized extraction conditions, and the calibration plot was linear in the concentration range of 0.30-200 ng/mL. With relatively low limit of detection and good precisions, the proposed method has the potential for the extraction and determination of methadone in biological samples.

An investigation into the polylactic acid texturization through thermomechanical processing and the improved d33 piezoelectric outcome of the fabricated scaffolds.

The bio/sensors performance has been established to be significantly affected through partially or entirely alignment of nano/microfibrous in polymeric mats. The matter of crystalline/amorphous proportion in semicrystalline polymers is another factor that can affect the application of the piezoelectric patches. The present work deals with fabricating the scaffolds of micro/nanofibers through a modified electrospinning procedure. The ratio of the relevant organic and polar solvents, the beading, the degree of fiber alignment, and fiber thickness have been intentionally elaborated. An unaligned unbeaded nanofibrous mat has been fabricated after tuning the solvents to poly-lactic acid ratio. This paper, for the first time, deals with the calculation of the value of d33 value of a commercial PLA and its improvement, it has been revealed that the d33 piezoelectric property is improved as a consequence of the thermo-mechanical processing above the cold crystallization temperature. The applied thermo (mechanical) processing causes the structural evolution from amorphous to crystallized states. Formation of the α and α' crystalline phases is introduced as the main responsible for the improvement of the piezoelectric property. This improvement not only is correlated with the degree of crystallinity, but also the orientation and alignment of the crystallites is known to be influential. In this respect, the complex helical chain structural evolution of poly-lactic acid has been analyzed through Herman's orientation function. It has been found that, besides the characterized disorder-to-order phase transformation, the C=O branched out dipoles interactions significantly affects by the texturization of the aligned polymeric chains in the direction of the electrospinning which is known as the main factor to promote the piezoelectric property of processed mat.

The effect of nano-size second precipitates on the structure, apatite-inducing ability and in-vitro biocompatibility of Ti-29Nb-14Ta-4.5Zr alloy.

The apatite formation and in-vitro biocompatibility of Ti-29Nb-14Ta-4.5Zr (TNTZ) alloy reinforced by various nano-sized phases of α″, α, and ω in the ß matrix have been studied. The electrochemical performances of the elaborated microstructures have been assessed through potentiodynamic polarization in the simulated body fluid (SBF) and interestingly, the ß + ω specimen exhibited an extraordinary corrosion resistance compared to the others. This was attributed to the uniform distribution, spherical morphology and coherent interface of the ω nano-precipitates. The polarization tests in simulated body fluid showed the high tendency of apatite formation on the surface of the ß- matrix contained ω precipitates. The in-vitro cytotoxicity analysis employing MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay showed >85% cell viability of the TNTZ alloy reinforced by nano-ω precipitations. Since this specimen showed the highest cell adhesion as well, it introduces this structure as a promising high potential candidate for biomedical applications due to its high corrosion resistance, biocompatibility, ultra-low cytotoxicity, and good cell adhesion.

Electromembrane-surrounded solid-phase microextraction coupled to ion mobility spectrometry for the determination of nonsteroidal anti-inflammatory drugs: A rapid screening method in complicated matrices.

A new robust method of electromembrane-surrounded solid-phase microextraction coupled to ion mobility mass spectrometry was applied for nonsteroidal anti-inflammatory drugs determination in complex matrices. This is the first time that a graphene/polyaniline composite coating is applied in electromembrane-surrounded solid-phase microextraction method. The homemade graphene/polyaniline composite is characterized by a high electrical conductivity and thermal stability. The variables affecting electromembrane-surrounded solid-phase microextraction, including extraction time; applied voltage and pH were optimized through chemometric methods, central composite design, and response surface methodology. Under the optimized conditions, limits of detection of 0.04 and 0.05 ng/mL were obtained for mefenamic acid and ibuprofen, respectively. The feasibility of electromembrane-surrounded solid-phase microextraction followed by ion mobility mass spectrometry was successfully confirmed by the extraction and determination of low levels of ibuprofen and mefenamic acid in human urine and plasma samples and satisfactory results were obtained.

Electromembrane extraction combined with cyclodextrin-modified capillary electrophoresis for the quantification of trimipramine enantiomers.

A sensitive, simple and reproducible method was developed for preconcentration and determination of trimipramine (TPM) enantiomers in biological samples using electromembrane extraction combined with cyclodextrin-modified capillary electrophoresis (CE). During the extraction, TPM enantiomers migrated from a 5 mL sample solution through a thin layer of 2-nitrophenyl octyl ether NPOE immobilized in the pores of a hollow fiber, and into a 20 µL acidic aqueous acceptor phase presented inside the lumen of the fiber. A Box-Behnken design and the response surface methodology (RSM) were used for the optimization of different variables on extraction efficiency. Optimized extraction conditions were: NPOE as supported liquid membrane, inter-electrode distance of 5 mm, stirring rate of 1000 rpm, 51 V potential difference, 34 min as the extraction time, acceptor phase pH 1.0 and donor phase pH 4.5. Then, the extract was analyzed using optimized cyclodextrin (CD)-modified CE method for the separation of TPM enantiomers. Best results were achieved using 100 mM phosphate running buffer (pH 2.0) containing 10 mM α-CD as the chiral selector, applied voltage of 18 kV and 20°C. The range of quantitation for both enantiomers was 20-500 ng/mL. The method was very reproducible so that intra- and interday RSDs (n=6) were <6%. The limits of quantitation and detection for both enantiomers were 20 and 7 ng/mL, respectively. Finally, this method was successfully applied to determine the concentration of TPM enantiomers in plasma and urine samples without any pre-treatment.

Optimization of carrier-mediated three-phase hollow fiber microextraction combined with HPLC-UV for determination of propylthiouracil in biological samples.

Carrier-mediated three-phase hollow fiber microextraction combined with high-performance liquid chromatography-ultra violet detection (HPLC-UV) was applied for the extraction and determination of propylthiouracil in biological samples. Propylthiouracil (PTU) was extracted from 7.5 mL of the basic solution (the source phase) with pH 12 into an organic phase (n-octanol containing 6% (w/v) of Aliquat 336 as the carrier) impregnated in the pores of a hollow fiber, and finally was back extracted into 24 µL of the acidic solution located inside the lumen of the hollow fiber (the receiving phase). The extraction was performed through the gradient of counter ion from the source to the receiving phase. The effects of different variables on the extraction efficiency were studied simultaneously using an experimental design. A half-fractional factorial design was employed for screening to determine the variables significantly affecting the extraction efficiency. Then, the factors with significant effect were optimized using a central composite design (CCD) and the response surface equations were developed. The optimal experimental conditions obtained from this statistical evaluation included: source phase, pH 12; temperature, 25°C; extraction time, 40 min; counter ion concentration, 2 mol L(-1) of NaClO(4); organic solvent 6% of Aliquat in octanol and without salt addition in the source phase. Under the optimized conditions, the preconcentration factors were between 125 and 198 and also the limit of detections (LODs) ranged from 0.1 µg L(-1) to 0.4 µg L(-1) in different biological samples. The calibration curve was linear (r(2)=0.998) in the concentration range of 0.5-1000 µg L(-1). Finally, the feasibility of the proposed method was successfully confirmed by extraction and determination of PTU in human plasma and urine as well as the bovine milk and meat samples in microgram per liter, and suitable results were obtained (RSDs<6.3%).

Molecular-imprinted polymer extraction combined with dispersive liquid-liquid micro-extractionfor ultra-preconcentration of mononitrotoluene.

Molecular-imprinted polymer (MIP) combined with dispersive liquid-liquid micro-extraction (DLLME) were developed for ultra-preconcentration and determination of mononitrotoluenes in wastewater samples using gas chromatography-flame ionization detector. MIP was synthesized by copolymerization of methacrylic acid-ethylene glycole dimethacrylate-2,2-azobisisobutyronitrile as the initiator that imprinted with 3-nitrotoluene as the template molecule. Effects of several factors, such as type and volume of eluent, adsorption, and desorption times of the analyte on the polymer, and breakthrough volume were investigated. Optimization of 3-nitrotoluene extraction from MIP was studied forward followed by DLLME. Preconcentration factor of MIP-DLLME method was about 2800 under the optimum conditions. The LOD of the proposed method was 0.02 µg/L and a linear dynamic range in the range of 0.04-20 µg/L was obtained. The performance of the present method was evaluated for extraction and determination of nitrotoluene compounds in wastewater samples in the range of microgram per liter and satisfactory results were achieved (RSD<13%).