Publications by authors named "Masoud Torkzadeh-Mahani"

64 Publications

Bioactive anti-oxidative polycaprolactone/gelatin electrospun nanofibers containing selenium nanoparticles/vitamin E for wound dressing applications.

J Biomater Appl 2021 Mar 15:8853282211001359. Epub 2021 Mar 15.

Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Kerman University of Medical Sciences, Kerman, Iran.

In this study, polycaprolactone/gelatin (PCL/GEL) electrospun nanofibers containing biogenic selenium nanoparticles (Se NPs) and Se NPs/vitamin E (VE) with average diameters of 397.8 nm and 279.5 nm, respectively (as determined by SEM inspection) were prepared and their effect on wound healing was evaluated using in-vivo studies. The energy dispersive X-ray (EDX) mapping, TEM micrograph, and FTIR spectra of the prepared nanofibers strongly demonstrated well entrapment of Se NPs and VE into scaffolds. An amount of 57% Se NPs and 43% VE were gradually released from PCL/GEL/Se NPs/VE scaffold after 4 days immersion in PBS solution (pH 7.4). The both PCL/GEL/Se NPs and PCL/GEL/Se NPs/VE scaffolds supported 3T3 cell proliferation and attachment as confirmed by MTT assay and SEM imaging. Complete re-epithelialization, low level of edema and inflammatory cells in coordination with high level of oriented collagens demonstrated the wound healing activity of PCL/GEL/Se NPs/VE. Besides, significant antioxidant efficacy of PCL/GEL/Se NPs and PCL/GEL/Se NPs/VE scaffolds was demonstrated according to GSH and MDA assays. To sum up, the prepared PCL/GEL/Se NPs/VE scaffold in the present study represented suitable healing effect on animal model which candidate it for further studies.
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http://dx.doi.org/10.1177/08853282211001359DOI Listing
March 2021

Niosomal virosome derived by vesicular stomatitis virus glycoprotein as a new gene carrier.

Biochem Biophys Res Commun 2021 01 31;534:980-987. Epub 2020 Oct 31.

Herbal and Traditional Medicines Research Center, Kerman University of Medical Sciences, Kerman, Iran; Department of Biochemistry, Faculty of Medicine, Kerman University of Medical Sciences, Kerman, Iran. Electronic address:

Virosomes as membranous vesicles with viral fusion protein in their membrane are versatile vehicles for cargo delivery. The vesicular stomatitis virus glycoprotein (VSV-G) is a common fusogenic protein used in virosome preparation. This glycoprotein has been used in liposomal systems so far, but in this study, we have tried to use the niosomal form instead of liposome for. Niosomes are vesicular systems composed of non-ionic surfactants. Niosomes were constructed by the thin-film hydration method. VSV-G gene in pMD2.G plasmid was expressed in the HEK293T cell line and then was reconstituted in the niosome bilayer. The formation of niosomal virosomes was confirmed with different methods such as SDS-PAGE gel, western blotting, and transmission electron microscopy (TEM). The efficiency of niosomal virosome was investigated with the pmCherry reporter gene. SDS-PAGE and western blotting proved the expression and successful insertion of protein into the bilayer. The TEM images showed the spike projection of VSV-G on the surface of niosomes. The transfection results showed high efficiency of niosomal virosomes as a novel carrier. This report has verified that niosome could be used as an efficient bilayer instead of liposome to construct virosomes.
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http://dx.doi.org/10.1016/j.bbrc.2020.10.054DOI Listing
January 2021

New insight into the molecular mechanism of the trehalose effect on urate oxidase stability.

J Biomol Struct Dyn 2020 Oct 1:1-11. Epub 2020 Oct 1.

Department of Biotechnology, Institute of Science, High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman-Iran, Iran.

Urate oxidase (EC 1.7.3.3) is a key enzyme in the purine metabolism which is applied in the treatment of gout and also, as a diagnostic reagent for uric acid detection. In the current study, the trehalose (TRE) effects as an additive on the structural stability and function of uricase were investigated. For recombinant expression of UOX in E. coli BL21 cells, firstly the coding sequence was subcloned into the pET-28a vector and after induction with IPTG, the recombinant UOX was purified by affinity chromatography using a Ni-NTA agarose column. To specify the trehalose effects on the urate oxidase (UOX) structure, optimum pH, optimum temperature, kinetic and thermodynamic parameters and also, the intrinsic fluorescence of UOX in the absence and presence of trehalose were examined. The UOX half-life is 24.32 min at 40 °C, whereas the UOX-TRE has a higher half-life (32.09 min) at this temperature. Generally, our findings confirm that trehalose has a protective effect on the enzyme structure. Optimum pH and temperature were 9 and 25 °C, respectively for both the naked and treated enzymes and their activity retained 42.18 and 64.80%, respectively after 48 h of incubation at room temperature. Also, theoretical results indicate that the random coil of the enzyme was converted to α-helix and β-sheet in the presence of trehalose which may preserve the integrity of the active site of the enzyme and increased the enzymatic activity. The MD simulation results indicated greater stability of the uricase structure in the presence of trehalose. Communicated by Ramaswamy H. Sarma.
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http://dx.doi.org/10.1080/07391102.2020.1828167DOI Listing
October 2020

Comprehensive Evaluation of Gene Expression in Negative and Positive Trigger-based Targeting Niosomes in HEK-293 Cell Line.

Iran J Pharm Res 2020 ;19(1):166-180

Physiology Research Center, Institute of Basic and Clinical Physiology Sciences, Kerman University of Medical Sciences, Kerman, Iran.

An efficient gene delivery system has some critical factors that enhance the efficiency of nanocarrier. These factors are low production cost, high bioavailability, high encapsulation efficiency, controllable release, and targeting ability. Niosome (the nonionic surfactant vesicles) was considered as a promising gene delivery system. Niosome can increase stability and uptake of active agents. We used all mentioned factors in one optimized formulation entitled plasmid- loaded magnetic niosomes (PMN). To increase the bioavailability of niosomes, we used ergosterol (a natural lipid) instead of cholesterol in structure of niosome. Also, cetyl trimethyl ammonium bromide (CTAB) in different concentrations was used to improve encapsulation of plasmid and compared to niosomes that did not have CTAB (negative niosome). Afterward, magnetic nanoparticle (FeO@SiO) was synthesized and loaded into niosome to obtain targeting ability. Prepared formulations were evaluated regarding size, zeta potential, morphology, encapsulation of magnetic nanoparticles and plasmid (Pm-cherry-N1), release rate, and transfection efficiency. Results demonstrated that optimum formulation (Nio/CTAB3%/Fe/P) has a nanometric size (118 ± 2.31 nm, positive zeta potential (+25 ± 0.67 mV), high loading of plasmid (72%), and good gene expression (35%). Interestingly, after applying a magnetic field below the cell plate, we obtained ac increased gene expression from 35% to 42%. These results showed that this new formulation would have a promising future and also can be used for delivering the other drugs and active agents.
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http://dx.doi.org/10.22037/ijpr.2019.112058.13507DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7462495PMC
January 2020

A new formulation of hydrophobin-coated niosome as a drug carrier to cancer cells.

Mater Sci Eng C Mater Biol Appl 2020 Aug 18;113:110975. Epub 2020 Apr 18.

Physiology Research Center, Institute of Basic and Clinical Physiology Sciences, Kerman University of Medical Sciences, Kerman, Iran; Department of Biochemistry, School of Medicine, University of Medical Sciences, Kerman, Iran.

Hydrophobin-1 (HFB-1) found on the surface of fungal spores, plays a role in the lack of antigen recognition by the host immune system. The present study aimed to evaluate the potential application of HFB-1 for the delivery of doxorubicin (Dox) into different cell lines. Coating the surface of niosomes (Nio) with HFB-1 leads to the hypothesis that this protein can confer protection against in vivo immune-system recognition and prevent the immune response. Thus, HFB-1 could become a promising alternative to polyethylene glycol (PEG). Here, HFB-1-coated niosome loaded with doxorubicin (Dox) based on Span 40, Tween 40 and cholesterol was prepared and compared with the PEG-coated niosome. Physicochemical characteristics of the prepared formulations in terms of size, zeta potential, polydispersity index (PDI), morphology, entrapment efficiency (EE), and release rate were evaluated at different pH levels (2, 5.2, and 7.4). In the end, the in vitro cytotoxicity assay was performed on four different cancer cell lines namely A549, MDA-MB-231, C6 and PC12 in addition to one control cell line (3 T3) to ensure the formulation's selectivity against cancer cells. Results showed that the niosomes coated with HFB-1 presented better size distribution, higher EE, more sustained release profile, enhanced biocompatibility and improved anticancer effects as compared to the PEG-coated niosomes. Interestingly, the viability percentage of the control cell line was higher than different cancer cells when treated with the formulations, which indicates the higher selectivity of the formulation against cancer cells. In conclusion, loading the niosomes with Dox and coating them with HFB-1 enhanced their efficacy and selectivity toward cancer cells, presenting a promising drug delivery system for sustained drug release in cancer treatment.
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http://dx.doi.org/10.1016/j.msec.2020.110975DOI Listing
August 2020

A label-free aptasensor for highly sensitive detection of homocysteine based on gold nanoparticles.

Bioelectrochemistry 2020 Aug 4;134:107497. Epub 2020 Mar 4.

Research Center for Tropical and Infectious Diseases, Kerman University of Medical Sciences, Kerman, Iran.

In the present study, an original electrode fabrication approach was devised to create a label free sensitive electrochemical aptasensor for the detection of Homocysteine (Hcy) (Homocysteine signal was used for detection). To bind certain targets, synthetic oligonucleotides used as aptamers (APs) were specifically selected. Aptamers are substitutes for antibodies for analytical devices because of their sensitivity and high affinity. In this study, Hcy-Binding-Aptamer (HBA) was grafted onto the surface of Au nanoparticles/Glassy Carbon Electrode (Au/GCE) in order to create an aptasensor. The effects of buffer concentration, buffer type, interaction time, and aptamer concentration were investigated and optimized. In addition, Differential Pulse Voltammetry (DPV) was implemented to identify homocysteine. Favorable performance was achieved at a detection limit of 0.01 μM (S/N = 3) and linear range 0.05-20.0 μM. Furthermore, the fabricated aptasensor displayed desirable stability and reproducibility. The developed electrochemical aptasensor was found to have reasonable selectivity for the detection of homocysteine in the presence of cysteine and methionine. Analysis of real samples showed good ability of the proposed homocysteine biosensor to provide sensitive, quick, easy, and cost effective measurement of homocysteine in human blood serum and urine samples.
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http://dx.doi.org/10.1016/j.bioelechem.2020.107497DOI Listing
August 2020

Comparative transcriptome provides molecular insight into defense-associated mechanisms against spider mite in resistant and susceptible common bean cultivars.

PLoS One 2020 4;15(2):e0228680. Epub 2020 Feb 4.

Department of Agronomy and Plant Breeding, Faculty of Agriculture, University of Tehran, Karaj, Iran.

Common bean (Phaseolus vulgaris L.) is a major source of proteins and one of the most important edible foods for more than three hundred million people in the world. The common bean plants are frequently attacked by spider mite (Tetranychus urticae Koch), leading to a significant decrease in plant growth and economic performance. The use of resistant cultivars and the identification of the genes involved in plant-mite resistance are practical solutions to this problem. Hence, a comprehensive study of the molecular interactions between resistant and susceptible common bean cultivars and spider mite can shed light into the understanding of mechanisms and biological pathways of resistance. In this study, one resistant (Naz) and one susceptible (Akhtar) cultivars were selected for a transcriptome comparison at different time points (0, 1 and 5 days) after spider mite feeding. The comparison of cultivars in different time points revealed several key genes, which showed a change increase in transcript abundance via spider mite infestation. These included genes involved in flavonoid biosynthesis process; a conserved MYB-bHLH-WD40 (MBW) regulatory complex; transcription factors (TFs) TT2, TT8, TCP, Cys2/His2-type and C2H2-type zinc finger proteins; the ethylene response factors (ERFs) ERF1 and ERF9; genes related to metabolism of auxin and jasmonic acid (JA); pathogenesis-related (PR) proteins and heat shock proteins.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0228680PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6999899PMC
May 2020

Bimetallic nickel-ferrite nanorod particles: greener synthesis using rosemary and its biomedical efficiency.

Artif Cells Nanomed Biotechnol 2020 Dec;48(1):242-251

NanoBioElectrochemistry Research Center, Bam University of Medical Sciences, Bam, Iran.

Nickel-ferrite (NiFeO) nanorods particles (NRP) was biosynthesised for the first time by the Rosemary Extract. The NRP was fully characterised, including the type, nanostructure and physicochemical properties of using XRD, HRTEM, FeSEM, XPS, FTIR and VSM. TEM confirmed rod-shaped nano-sized particles with average sizes ranging from 10 nm to 28 nm. The EDAX Analysis showed the presence of iron, nickel, oxygen, and carbon. XRD analysis confirmed the synthesis of NiFeO crystals. XPS curves showed photoelectron for iron, oxygen and nickel. EDS showed the atomic, weight percentages ratios of Ni(12%): Fe(24%) and: O(48) are close to the theoretical value (Ni: Fe: = 1:2:4), of bimetallic magnetic NiFeO NRP. NiFeO NRP had cytotoxicity effect on MCF-7 cells survival which suggests that NiFeO NRP can be used as a new class of anticancer agent in design novel cancer therapy research.
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http://dx.doi.org/10.1080/21691401.2019.1699830DOI Listing
December 2020

The Blepharis persica seed hydroalcoholic extract synergistically enhances the apoptotic effect of doxorubicin in human colon cancer and gastric cancer cells.

Mol Biol Rep 2020 Feb 17;47(2):843-853. Epub 2019 Dec 17.

Faculty of Basic Sciences, Department of Plant Biology, University of Jiroft, 8th km Bandar Abbas Road, Kerman, Iran.

The goal of this survey is to evaluate the anti-proliferative effects of the hydroalcholic extract of Blepharis persica seeds and its synergic effect on doxorubicin (DOX) in human colon cancer (HT-29) and gastric cancer cell (AGS) lines. 70% Ethanol was used for extraction of B. persica seed. Aluminum-chloride colorimetric and Folin-Ciocalteu reagent methods were used to measure total flavonoid and total phenolic contents of the extract respectively. Gas chromatography-mass spectrometry (GC-MS) analysis of the B. persica extract was performed on GC-MS equipment after silylation. HT-29, AGS, and human fibroblast (SKM) cell lines were treated by different concentration of the B. persica extract, (DOX) and the combination of extraction and DOX. The cytotoxicity was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay while the apoptosis induction was monitored using flowcytometry by annexin-V FITC/PI double-staining. The changes in expression levels of BAX and BCL-2 were determined using Real-Time RT-qPCR. GC-MS analysis of the hydroalcoholic extract from B. persica seeds revealed 24 major components. The MTT assay revealed the cytotoxicity against three cell lines and also it was shown that 125 ng/mL of DOX and 0.625 mg/mL of B. persica extract had synergistic behavior against HT29 cell line. These results showed B. persica extract induced apoptosis in AGS and HT29 cells and its extract caused dose-dependent increase in up-regulation of BAX level (p < 0.05) and down-regulation of BCL (p < 0.05). B. persica showed the synergistic effect in combination with DOX on HT29 cell line. These findings demonstrated a basis for further studies on the characterization and mechanistic evaluation of the bioactive compounds of B. persica extract which had antiproliferative effects on cancer cell lines.
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http://dx.doi.org/10.1007/s11033-019-04711-zDOI Listing
February 2020

From in vitro to in silico: Modeling and recombinant production of DT-Diaphorase enzyme.

Int J Biol Macromol 2020 Jan 5;143:213-223. Epub 2019 Dec 5.

Department of Biotechnology, Institute of Science, High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran. Electronic address:

DT-Diaphorase (DTD) belonging to the oxidoreductase family, is among the most important enzymes and is of great significance in present-day biotechnology. Also, it has potential applications in glucose and pyruvate biosensors. Another important role of the DTD enzyme is in the detection of Phenylketonuria disease. According to the above demands, at first, we tried to study molecular cloning and production of recombinant DTD in E. coli BL21 strain. We have successfully cloned, expressed, and purified functionally active diaphorase. The amount of enzyme was increased in 10-h using IPTG induction, and the recombinant protein was purified by Ni-NTA agarose affinity chromatography. After that, the kinetic and thermodynamic parameters of the enzyme, optimum temperature and pH were also investigated to find more in-depth information. In the end, to represent the connections between the structures and function of this enzyme, the molecular dynamics simulations have been considered at two temperatures in which DTD had maximum and minimum activity (310 and 293 K, respectively). The results of MD simulations indicated that the interaction between NADH with phenylalanine 232 residue at 310 K is more severe than other residues. So, to investigate the interaction details of NADH/PHE 232 the DFT calculations were done.
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http://dx.doi.org/10.1016/j.ijbiomac.2019.12.029DOI Listing
January 2020

cytotoxicity assay of D-limonene niosomes: an efficient nano-carrier for enhancing solubility of plant-extracted agents.

Res Pharm Sci 2019 Oct 4;14(5):448-458. Epub 2019 Oct 4.

Department of Biotechnology, Institute of Science, High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, I.R. Iran.

The low solubility of the plant-extracted agent like D-limonene in cancer therapy is a critical problem. In this study, we prepared D-limonene-loaded niosomes (D-limonene/Nio) for cancer therapy through cytotoxicity assay of HepG2, MCF-7, and A549 cell lines. The niosomal formulation was prepared by film hydration technique with Span 40: Tween 40: cholesterol (35:35:30 molar ratio) and characterized for vesicle distribution size, morphology, entrapment efficiency (EE%), and release behaviour. The obtained niosomes showed a nanometric size and spherical morphology with EE% about 87 ± 1.8%. Remarkably prolonged release of D-limonene from niosomes compared to free D-limonene observed. The loaded formulation showed significantly enhanced cytotoxic activity with all three cancer cell lines (HepG2, Macf-7 and A549) at the concentration of 20 μM. These results indicated that niosome loaded with phytochemicals can be a promising nano-carrier for cancer therapy applications.
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http://dx.doi.org/10.4103/1735-5362.268206DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6827193PMC
October 2019

Electrochemical aptasensor for activated protein C using a gold nanoparticle - Chitosan/graphene paste modified carbon paste electrode.

Bioelectrochemistry 2019 Dec 25;130:107322. Epub 2019 Jun 25.

Department of Biotechnology, Institute of Science, High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran. Electronic address:

In this work, a selective and simple electrochemical aptasensor was developed for the detection of activated protein C by employing methylene blue (MB) as a redox indicator. An activated protein C aptamer (APC-apt) was covalently immobilized on the surface of a carbon paste electrode modified with gold nanoparticle - chitosan /graphene paste (AuNPs-Chi/Gr). The AuNPs-Chi/Gr paste increased electrochemical peak current and immobilized the aptamer on the electrode surface. The process of aptasensor construction and successful immobilization of the aptamer on the electrode surface was confirmed by electrochemical cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Differential pulse voltammetry (DPV) was used to determine the methylene blue peak current. By replacing APC instead of MB at the electrode surface, the cathodic current of the MB decreases, and this decrease corresponds to the APC concentration. Under optimum conditions, the APC concentration was detected in the range from of 0.1 ng·mL to 40 μg·mL with a relatively low detection limit of 0.073 ng·mL. This method was then applied to the determination of APC in human serum samples. The results revealed that this strategy can be used to measure other proteins in biological samples.
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http://dx.doi.org/10.1016/j.bioelechem.2019.06.007DOI Listing
December 2019

A hydrophobin-based-biosensor layered by an immobilized lactate dehydrogenase enzyme for electrochemical determination of pyruvate.

Bioelectrochemistry 2019 Dec 26;130:107323. Epub 2019 Jun 26.

Department of Biotechnology, Institute of Science, High Technology and Environmental Science, Graduate University of Advanced Technology, Kerman, Iran. Electronic address:

Appropriate enzyme immobilization on the electrode surface in order to access its active site has always been an important strategy for electrode modification. In this report, lactate dehydrogenase enzyme was appropriately immobilized on the glassy carbon electrode via hydrophobin (HFB1) and graphene oxide nanocomposite. The step-by-step modification was successfully confirmed by water contact analysis, cyclic voltammetry, and electrochemical impedance spectroscopy. Under optimum conditions, this biosensor demonstrated a detection limit of 8.69 nM and RSD of 4.3% and 3.6% (n = 5) for reproducibility and repeatability. The effect of scan rate on the oxidation behavior of NADH was investigated by cyclic voltammetry; and diffusion coefficient for NADH was estimated at 6.27 × 10 cm.s. The apparent Michaelis-Menten constant (K) was amperometrically determined and it was lower than K for the free enzyme. Also, the modified electrode represented good stability after nine days with 6% decrease in current. The proposed assay was successfully used in real sample-serum-analysis and the obtained recoveries were between 93% and 104.0%.
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http://dx.doi.org/10.1016/j.bioelechem.2019.06.008DOI Listing
December 2019

Diosgenin-loaded niosome as an effective phytochemical nanocarrier: physicochemical characterization, loading efficiency, and cytotoxicity assay.

Daru 2019 Jun 27;27(1):329-339. Epub 2019 May 27.

Department of Biotechnology, Institute of Science, High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran.

Background: The use of phytochemicals to prevent or suppress tumours is known as chemoprevention. Numerous plant-derived agents have been reported to have anticancer potentials. As one such anticancer phytochemical, diosgenin has several applications which are nevertheless limited due to its low solubility in water.

Methods: We loaded diosgenin into niosome to increase its solubility and hence efficiency. Diosgenin-niosome (diosgenin loaded into niosome) was prepared by thin-film hydration method and characterised by optical microscopy, dynamic light scattering (DLS), scanning electron microscopy (SEM), and UV-visible spectrophotometry. Also, loading efficiency, in vitro drug release, and cytotoxicity assay were performed on HepG2 cell line.

Results And Discussion: Diosgenin-niosome has a nanometric size with a normal size distribution and spherical morphology. The loading efficiency of diosgenin was about 89% with a sustainable and controllable release rate. Finally, the viability of free diosgenin was 61.25%, and after loading into niosomes, it was improved to 28.32%.

Conclusion: The results demonstrated that niosomes increase the solubility of naturally derived hydrophobic chemicals and thus enhance their anticancer effect. Graphical abstract.
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http://dx.doi.org/10.1007/s40199-019-00277-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6592984PMC
June 2019

Evaluation of Carum-loaded Niosomes on Breast Cancer Cells:Physicochemical Properties, In Vitro Cytotoxicity, Flow Cytometric, DNA Fragmentation and Cell Migration Assay.

Sci Rep 2019 05 9;9(1):7139. Epub 2019 May 9.

Department of Biotechnology, Institute of Science, High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran.

Thymoquinone (TQ), a phytochemical compound found in Carum carvil seeds (C. carvil), has a lot of applications in medical especially cancer therapy. However, TQ has a hydrophobic nature, and because of that, its solubility, permeability and its bioavailability in biological mediums are poor. To diminish these drawbacks, we have designed a herbal carrier composed of Ergosterol (herbal lipid), Carum carvil extract (Carum) and nonionic surfactants for herbal cancer treatment. C. carvil was extracted and characterized by GC/Mass. Two different formulations containing TQ and Carum were encapsulated into niosomes (Nio/TQ and Nio/Carum, respectively) and their properties were compared together. Morphology, size, zeta potential, encapsulation efficiency (EE%), profile release rate, in vitro cytotoxicity, flow cytometric, DNA fragmentation and cell migration assay of formulations were evaluated. Results show that both loaded formulations have a spherical morphology, nanometric size and negative zeta potential. EE% of TQ and Carum loaded niosomes was about 92.32% ± 2.32 and 86.25% ± 1.85, respectively. Both loaded formulations provided a controlled release compared with free TQ. MTT assay showed that loaded niosomes have more anti-cancer activity compared with Free TQ and free Carum against MCF-7 cancer cell line and these results were confirmed by flow cytometric analysis. Cell cycle analysis showed G2/M arrest in TQ, Nio/TQ and Nio/Carum formulations. TQ, Nio/TQ and Nio/Carum decreased the migration of MCF7 cells remarkedly. These results show that the TQ and Carum loaded niosomes are novel carriers with high efficiency for encapsulation of low soluble phytochemicals and also would be favourable systems for breast cancer treatment.
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http://dx.doi.org/10.1038/s41598-019-43755-wDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6509162PMC
May 2019

cytotoxicity studies of parent and nanoencapsulated Holmium-2,9-dimethyl-1,10-phenanthroline complex toward fish-salmon DNA-binding properties and antibacterial activity.

J Biomol Struct Dyn 2019 10 16;37(17):4437-4449. Epub 2019 Jan 16.

Department of Biotechnology, Institute of Science, High Technology & Environmental Science, Graduate University of Advance Technology , Kerman , Iran.

In this study, the interaction of Holmium (Ho) complex including 2, 9-dimethyl-1,10-phenanthroline, also called Neocuproine (Neo), [Ho(Neo)Cl.HO], as fluorescence probe with fish-salmon DNA (FS-DNA) is studied during experimental investigations. Multi-spectroscopic methods are utilized to determine the affinity binding constants () of complex-FS-DNA. It is found that fluorescence of Ho complex is strongly quenched by the FS-DNA through a static quenching procedure. Under optimal conditions in Tris(trishydroxymethyl-aminomethane)-HCl buffer at 25 °C with pH ≈ 7.2, intrinsic binding constant of Ho complex is 6.12 ± 0.04 × 10 M. Also, the binding site number and Stern-Volmer quenching constant are calculated. There are different approaches, including iodide quenching assay, salt effect and thermodynamical assessment to determine the features of the binding mode between Ho complex and FS-DNA. Also, the parent and starch and lipid nanoencapsulated Ho complex, as potent antitumor candidates, were synthesized. The main structure of Ho complex is maintained after encapsulation using starch and lipid nanoparticles. 3-[4,5-Dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT) method was used to assess the anticancer properties of Ho complex and its encapsulated forms on human cancer cell lines of human lung carcinoma cell line and breast cancer cell line. In conclusion, these compounds could be considered as new antitumor candidates. Communicated by Ramaswamy H. Sarma.
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http://dx.doi.org/10.1080/07391102.2018.1557077DOI Listing
October 2019

Genomic and protein structure analysis of the luciferase from the Iranian bioluminescent beetle, Luciola sp.

Int J Biol Macromol 2019 Mar 28;124:689-698. Epub 2018 Nov 28.

Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences Shiraz, Iran.

To date, two Iranian luciferase genes from the Lampyris turkestanicus and Lampyroidea maculata have been carefully studied. Here, we report the cloning and characterization of the gene and protein of luciferase enzyme from the beetle of an Iranian lampyrid species, Luciola sp. (Coleoptera-Lampyridae). In this study, a Luciola sp. firefly was collected from the Yasouj area of Iran and its luciferase gene sequence was cloned and characterized. The genomic DNA length for this luciferase was the 1950 bp that combined of seven exons and separated by six introns. The results of multiple sequence alignment show that this gene has the most similarity with DNA gene luciferase from the Hotaria unmunsana species. Further analysis determined accurately the location of these introns in the luciferase gene. However, the deduced amino acid sequences of the luciferase gene (548 residues) showed that this luciferase had 97.8% resemblance to luciferase from Lampyroidea maculata species. By in silico modeling of firefly luciferase in an I-TASSER server, the 3D structure of this enzyme was evaluated. The results of phylogenetic tree analysis display the close evolutionary relationship of this luciferase gene and luciferase gene from the Lampyroidea maculata and Hotaria unmunsana.
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http://dx.doi.org/10.1016/j.ijbiomac.2018.11.264DOI Listing
March 2019

Controlled release of lawsone from polycaprolactone/gelatin electrospun nano fibers for skin tissue regeneration.

Int J Biol Macromol 2019 Mar 27;124:478-491. Epub 2018 Nov 27.

Textile Engineering Department, Amirkabir University of Technology, Tehran, Iran.

In this study, we aimed to evaluate the role of electrospun nanofibers containing lawsone (2-hydroxy-1,4-naphthoquinone) in wound healing. Different concentrations of lawsone (0.5%, 1%, 1.5%) were electrospun in polycaprolactone-gelatin (PCL/Gel) polymers in core-shell architecture. Nanofibers were characterized using scanning electron microscopy (SEM), Transmission electron microscopy (TEM). Coaxial electrospinning of PCL/Gel/lawsone (PCL/Gel/Law) scaffolds prolonged the release of lawsone over a period of 20 days. In vitro bioactivity of fibers on human gingiva fibroblast cells (HGF) was evaluated by MTT assay. The PCL/Gel/Law 1% increased cell attachment and proliferation significantly. Additionally, the in vitro gene expression of transforming growth factor β (TGF-B1), collagen (COL1) and epidermal growth factor (EGF) was monitored using RT-qPCR technique, which there was significant increase in TGF-B1 and COL1 gene expression in PCL/Gel/Law 0.5% and 1% mats. In vivo wound healing activity of lawsone loaded scaffolds in rat wound model revealed that the PCL/Gel/Law 1% had the highest impact on healing by increasing re-epithelialization of the wound after 14 days. It can be inferred that lawsone 1% incorporation in the core of PCL/Gel electrospun nanofibers has excellent characteristics and can be used as wound dressing patch in medicine.
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http://dx.doi.org/10.1016/j.ijbiomac.2018.11.237DOI Listing
March 2019

Synthesis and characterization of a novel organosilane-functionalized chitosan nanocarrier as an efficient gene delivery system: Expression of green fluorescent protein.

Int J Biol Macromol 2019 Mar 26;125:143-148. Epub 2018 Nov 26.

Department of New Materials, Institute of Science and High Technology and Environmental Sciences, Graduate University of Advanced Technology, PO Box 76315-117, Kerman, Iran. Electronic address:

Semi-essential arginine amino acid was selected to synthesis an organosilane linker for modifying chitosan biopolymer. The novel organosilane linker which was chemically synthesized by reaction of arginine with 3‑chloropropyl trimethoxy silane, covalently bonded to the chitosan amino group. The chemical structure of resulting nanocarrier was characterized by H NMR, wide-X-ray diffraction, TEM, FESEM and EDX. A maximum retardation capacity of the nanocarrier to the plox plasmid was obtained 3 at physiological pH (7.4). The mean of cell viability and cytotoxicity of the nanocarrier was determined 85% by MTT assay. In addition, the gene transfection of the nanocarrier was obtained top of 20% gene expression. The studies have shown very good DNase 1 enzyme protection of plasmid by the nanocarrier.
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http://dx.doi.org/10.1016/j.ijbiomac.2018.11.145DOI Listing
March 2019

Stabilization of d-lactate dehydrogenase diagnostic enzyme via immobilization on pristine and carboxyl-functionalized carbon nanotubes, a combined experimental and molecular dynamics simulation study.

Arch Biochem Biophys 2019 01 23;661:178-186. Epub 2018 Nov 23.

Department of Biotechnology, Institute of Science, High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran. Electronic address:

The most important mode of enzyme inactivation is thermal inactivation. Immobilization technology is an efficient approach to elongate the life-time of enzymes. d-lactate dehydrogenase (D-LDH) was stabilized at high temperatures with immobilization on CNT and fCNT. The kinetic and thermodynamic parameters, optimum temperature and pH, and the intrinsic fluorescence of free and immobilized enzymes were examined in the present study. Also, an attempt was made to investigate the effect of CNT and fCNT on the adsorption and conformation of d-lactate dehydrogenase using molecular dynamics (MD) simulations. In comparison with free enzyme, the immobilized enzyme displayed an improved stability at high temperatures and, therefore, the immobilized enzyme is suitable for use in the industry because most reactions in the industry happen at high temperatures. Results of the present study showed that the adsorption of enzyme on CNT is mediated through the van der Waals and π-π stacking interactions, whereas in the adsorption of enzyme on fCNT in addition to hydrophobic interactions, the hydrogen bonding between enzyme and functional groups of fCNT is involved. Moreover, RMSD, RMSF and secondary structure analysis indicate that the fCNT protects the conformation of enzyme more than CNT. Therefore, D-LDH can be efficiently immobilized upon the fCNT compared to the pristine CNT.
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http://dx.doi.org/10.1016/j.abb.2018.11.019DOI Listing
January 2019

In silico and in vitro study of magnetic niosomes for gene delivery: The effect of ergosterol and cholesterol.

Mater Sci Eng C Mater Biol Appl 2019 Jan 8;94:234-246. Epub 2018 Sep 8.

Department of Biochemistry, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran.

A low transfection efficiency and failure to deliver therapeutic genes to target organs limit the use of vesicular systems in gene therapy. In this study, magnetic niosomes were used to improve transfection efficiency and overcome limitations. In this light, Tween 60 and Span 60 molecules were employed as the bilayer component and ergosterol and/or cholesterol as membrane-stabilizing agents. We studied the structural and dynamical properties of cholesterol-containing niosomes (ST60/Chol) and ergosterol-containing vesicles (ST60/Ergo) using the molecular dynamics (MD) simulation technique. In in vitro experiments, the protamine-condensed DNA along with magnetic nanoparticles were prepared and incorporated into the niosome to form magnetic niosome-entrapped protamine-condensed DNA (M-NPD). The MD simulation comparison of two bilayers showed that the ST60/Ergo vesicles have better properties for gene delivery. Our in vitro results confirmed the in silico results and revealed that Ergo-niosomes have smaller size, better polydispersity, and slower release of plasmid than Chol-niosome. Moreover, M-NPD-Ergo showed higher cellular uptake and gene expresssion in HEK-293T cell line compared to M-NPD-Chol vesicles.
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http://dx.doi.org/10.1016/j.msec.2018.09.026DOI Listing
January 2019

Tris-chelated complexes of nickel(II) with bipyridine derivatives: DNA binding and cleavage, BSA binding, molecular docking, and cytotoxicity.

J Biomol Struct Dyn 2019 09 5;37(15):3887-3904. Epub 2019 Feb 5.

a Pharmaceutics Research Center, Institute of Neuropharmacology, Kerman University of Medical Sciences , Kerman , Iran.

Two nickel(II) complexes with substituted bipyridine ligand of the type [Ni(NN)](ClO), where NN is 4,4'-dimethyl-2,2'-bipyridine (dimethylbpy) () and 4,4'-dimethoxy-2,2'-bipyridine (dimethoxybpy) (), have been synthesized, characterized, and their interaction with DNA and bovine serum albumin (BSA) studied by different physical methods. X-ray crystal structure of shows a six-coordinate complex in a distorted octahedral geometry. DNA-binding studies of and reveal that both complexes sit in DNA groove and then interact with neighboring nucleotides differently; undergoes a partial intercalation. This is supported by molecular-docking studies, where hydrophobic interactions are apparent between and DNA as compared to hydrogen bonding, hydrophobic, and interactions between and DNA minor groove. Moreover, the two complexes exhibit oxidative cleavage of supercoiled plasmid DNA in the presence of hydrogen peroxide as an activator in the order of >. In terms of interaction with BSA, the results of spectroscopic methods and molecular docking show that binds with BSA only via hydrophobic contacts while interacts through hydrophobic and hydrogen bonding. It has been extensively demonstrated that the nature of the methyl- and methoxy-groups in ligands is a strong determinant of the bioactivity of nickel(II) complexes. This may justify the above differences in biomolecular interactions. In addition, the cytotoxicity of the complexes on human carcinoma cells lines (MCF-7, HT-29, and U-87) has been examined by MTT assay. According to our observations, and display cytotoxicity activity against selected cell lines. Communicated by Ramaswamy H. Sarma.
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http://dx.doi.org/10.1080/07391102.2018.1534700DOI Listing
September 2019

Magnetic delivery of antitumor carboplatin by using PEGylated-Niosomes.

Daru 2018 Sep 12. Epub 2018 Sep 12.

Department of Biotechnology, Institute of Science, High Technology & Environmental Sciences, Graduate University of Advanced Technology, Haft-Bagh Highway, Kerman, 7631133131, Iran.

To improve the efficiency of niosomal drug delivery, here we employed two tactics. First, niosomes were magnetized using FeO@SiO mangnetic nanoparticles, and second, their surface was modified by PEGylation. PEGylation was intended for increasing the bioavailability of niosomes, and magnetization was used for rendering them capable of targeting specific tissues. These PEGylated magnetic niosomes were also loaded with Carboplatin, an antitumor drug. Next, these niosomes were studied in terms of size, morphology, zeta potential, and drug entrapment efficiency. Then, the in vitro drug release from these modified niosomes was compared to that of both naked and nonmagnetized niosomes. Interestingly, although loading of naked-niosomes with magnetic particles lead to an increase in the rate of drug release, PEGylation of these magnetized niosomes caused a more sustained drug release. Thus, PEGylation of magnetic niosomes, besides improving their bioavailability, caused a slower and sustained release of the drug over time. Finally, studying the in vitro effectives of niosomal formulations towards MCF-7, a breast cancer cell line, showed that PEGylated magnetic niosomes had a satisfactory toxicity towards these cells in the presence of an external magnetic field. In conclusion, PEGylated magnetic niosomes showed enhanced qualities regarding the controlled release and delivery of drug. Graphical abstract ᅟ.
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http://dx.doi.org/10.1007/s40199-018-0215-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6154485PMC
September 2018

Lawsone-loaded Niosome and its antitumor activity in MCF-7 breast Cancer cell line: a Nano-herbal treatment for Cancer.

Daru 2018 Sep 27;26(1):11-17. Epub 2018 Jul 27.

Department of Biochemistry, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran.

Phytochemicals like Lawsone have some drawbacks that stem from their poor solubility. Low solubility in aqueous mediums results in low bioavailability, poor permeability and instability of phytochemical compounds in biological environments. The aim of this study was to design nanoniosomes containing Lawsone (Law) using non-ionic surfactants and cholesterol. Niosomes were prepared by thin film hydration method (TFH). Then, they were loaded with Henna extract (HLaw) and standard Lawsone (SLaw), and two resulted formulations were compared. The henna extract was analyzed by mass gas chromatography. Size, zeta potential, polydispersity index (PDI) and morphology of the loaded formulations were evaluated by dynamic light scattering (DLS) and scanning electron spectroscopy (SEM). The incorporation and release rate of Law from niosome bilayers were evaluated by UV-Vis spectroscopy. In vitro experiments were carried out to evaluate antitumor activity in MCF-7 cell line. The results showed distinct spherical shapes and particle sizes were about 250 nm in diameter and have negative zeta potentials. Niosomes were stable at 4 °C for 2 months. Entrapment efficiently of both formulations was about 70% and showed a sustained release profile. In vitro study exhibited that using of niosome to encapsulating Law can significantly increase antitumor activity of formulation in MCF-7 cell line compared to Law solution (free Law). Thus, niosomes are a promising carrier system for delivery of phytochemical compounds that have poor solubility in biological fluids. Graphical abstract ᅟ.
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http://dx.doi.org/10.1007/s40199-018-0207-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6154483PMC
September 2018

Improvement of kinetic properties and thermostability of recombinant Lepidium draba peroxidase (LDP) upon exposed to osmolytes.

Int J Biol Macromol 2018 Nov 8;119:1036-1041. Epub 2018 Aug 8.

Department of Biotechnology, Institute of Science and High technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran.

In this study, effects of different concentrations of glycine and D-sorbitol were analyzed on the activity and thermostability of recombinant Lepidium draba peroxidase (LDP). Based on the results, activity of the enzyme increased in the presence of various concentrations of these osmolytes. Maximum activity was detected for the enzyme in the presence of 300 mM glycine and 600 mM sorbitol. In presence of the aforementioned doses of osmolytes, enzyme affinity for substrate (3,3',5,5'-tetramethylbenzidine and HO) and V increased. According to the results, enzyme stability improved against temperature and HO. Furthermore, structural changes of the enzyme upon exposure to the osmolytes were revealed by the use of far-UV circular dichroism and fluorescence methods. The results showed, whereas the secondary structure of the enzyme was not significantly changed upon exposed to the osmolytes, the fluorescence studies revealed microenvironment of the aromatic residues dramatically affected by them. Overall, it may be speculated, structural changes of the enzyme upon exposed to the osmolytes, lead to the improvement of its kinetic properties and stability that can be benefit for using of it in in vitro applications.
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http://dx.doi.org/10.1016/j.ijbiomac.2018.08.021DOI Listing
November 2018

Double strand DNA-based determination of menadione using a FeO nanoparticle decorated reduced graphene oxide modified carbon paste electrode.

Bioelectrochemistry 2018 Dec 19;124:165-171. Epub 2018 Jul 19.

Department of Biotechnology, Institute of Science, High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran. Electronic address:

In this work an electrochemical label free DNA biosensor (ds-DNA) for the determination of menadione (MD) was developed. The biosensor was constructed using a modified nanocomposite consisting of FeO nanoparticles decorated reduced graphene oxide (Gr) on a carbon paste electrode (CPE). Scanning electron microscope (SEM), energy dispersive X-ray (EDAX) and Fourier transform infrared (FT-IR) spectroscopy confirmed the structure of the synthesized nanocomposites (electrode composition). The Gr-FeO nanocomposites formed a sensitive layer with large surface area. Electrochemical studies revealed that modification of the electrode surface with ds-DNA and Gr- FeO nanocomposite significantly increases the oxidation peak currents and reduces the peak potentials of MD. Under the optimum conditions, calibration curve was linear in the range of 0.3-100.0 nM with a detection limit of 0.13 nM. The relative standard deviation for 50.0 nM was 3.90% (n = 5). The proposed biosensor was successfully applied to the determination of MD.
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http://dx.doi.org/10.1016/j.bioelechem.2018.07.014DOI Listing
December 2018

Amperometric immunosensor for prolactin hormone measurement using antibodies loaded on a nano-Au monolayer modified ionic liquid carbon paste electrode.

Talanta 2018 Oct 15;188:701-707. Epub 2018 Jun 15.

Biotechnology Department, Institute of Science and High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran.

The fabrication of a novel electrochemical immunosensor for rapid and precise determination of prolactin was carried out using carbon paste electrode (CPE) consist of ionic liquid (IL) and graphite. Gold nanoparticles were employed as a modifier on the surface of CPE to immobilize the prolactin antibody (anti-PRL). The immunoassay was set up by sandwiching the antigen between prolactin antibody and the polyclonal anti-human-prolactin antibody labeled with horseradish peroxidase (HRP-labeled anti-PRL) as secondary antibody, on the surface of modified CPE. The reaction between O-aminophenol (OAP) and HO which is catalyzed by labeled HRP on the sandwich immunosensor generate a signal in differential pulse voltammetry (DPV) that is used to determine the concentration of prolactin. This immunosensor provides the measurement of prolactin concentration in a linear range of 25.0-2000.0 mIU L with a detection limit 12.5 mIU L. Moreover, it is applicable in the clinical assay of prolactin due to its high sensitivity and acceptable stability.
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http://dx.doi.org/10.1016/j.talanta.2018.06.044DOI Listing
October 2018

Optimization of in vitro refolding conditions of recombinant Lepidium draba peroxidase using design of experiments.

Int J Biol Macromol 2018 Oct 26;118(Pt B):1369-1376. Epub 2018 Jun 26.

Department of Nanotechnology Engineering, Mineral Industries Research Center, Shahid Bahonar University of Kerman, Kerman, Iran.

The main objective of this study was to optimize the in vitro refolding conditions of the recombinant Lepidium draba peroxidase (LDP). Initially, the effects of various factors were investigated on LDP refolding yield using one-factor-at-a-time (OFAT) method. Based on the OFAT results, optimum concentrations for LDP refolding were 2 M urea, 2 mM CaCl, 0.42 mM l-glutathione oxidized (GSSG), 0.20 mg/ml protein, and 12 μM hemin as well as pH 7. Secondly, according to the OFAT results, design of experiments (DOE) was applied for investigation of the interactions between factors including protein (P), urea (U), CaCl (C), and GSSG (G). The results showed the possible interaction between PC, PG, PU, and GU. Lastly, response surface methodology (RSM) was used for final refolding conditions optimization. The final optimized refolding conditions for LDP were conducted as 2 M urea, 1 mM CaCl, 0.70 mM GSSG, 0.07 mM DTT, 0.15 mg/ml protein which they obtained from RSM results and 12 μM hemin, and pH 7 according to the results of OFAT method. Overall, under optimal conditions, 23.4 mg active refolded LDP per liter of expression medium was obtained. So, the refolding yield was calculated to be approximately 48%.
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http://dx.doi.org/10.1016/j.ijbiomac.2018.06.122DOI Listing
October 2018

Synthesis and characterization of aminotetrazole-functionalized magnetic chitosan nanocomposite as a novel nanocarrier for targeted gene delivery.

Mater Sci Eng C Mater Biol Appl 2018 Aug 5;89:166-174. Epub 2018 Apr 5.

Department of New Materials, Institute of Science and High Technology and Environmental Sciences, Graduate University of Advanced Technology, PO Box 76315-117, Kerman, Iran. Electronic address:

In the present study, FeO/chitosan biopolymer grafted to a novel organosilane modified 5-amino-1H-tetrazole, a kind of drug intermediate, was successfully synthesized by chemical modification technique and evaluated as a high potential carrier of gene delivery. The loading capacity was evaluated, and in vitro release of nanocarrier was assessed using the dialysis method. The transfection efficiency of plasmid was optimal at an N/P ratio of 3. The chemically modified chitosan showed no inherent toxicity toward the cells. The synthesized nanocarrier had enhanced release of the plasmid at physiological pH 7.4. The N-functionalized magnetic chitosan nanocarrier demonstrated its efficacy in the enhancement of gene expression in the HECK-293T cell line. Therefore, the novel magnetic N-functionalized chitosan showed promise as a highly efficient gene carrier with potential applications in cancer therapy.
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http://dx.doi.org/10.1016/j.msec.2018.03.032DOI Listing
August 2018

Statistical optimization of kojic acid production by a UV-induced mutant strain of Aspergillus terreus.

Braz J Microbiol 2018 Oct - Dec;49(4):865-871. Epub 2018 Apr 24.

Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Kerman University of Medical Sciences, Kerman, Iran; Pharmaceutics Research Center, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran. Electronic address:

The ability of four Aspergillus strains for biosynthesis of kojic acid was evaluated among which Aspergillus terreus represented the highest level (2.21g/L) of kojic acid production. Improvement kojic acid production ability of A. terreus by random mutagenesis using different exposure time to ultraviolet light (5-40min) was then performed to obtain a suitable mutant of kojic acid production (designated as C, 7.63g/L). Thereafter, design of experiment protocol was employed to find medium components (glucose, yeast extract, KHPO (NH)SO, and pH) influences on kojic acid production by the C mutant. A 2 fractional factorial design augmented to central composite design showed that glucose, yeast extract, and KHPO were the most considerable factors within the tested levels (p<0.05). The optimum medium composition for the kojic acid production by the C mutant was found to be glucose, 98.4g/L; yeast extract, 1.0g/L; and KHPO, 10.3mM which was theoretically able to produce 120.2g/L of kojic acid based on the obtained response surface model for medium optimization. Using these medium compositions an experimental maximum Kojic acid production (109.0±10g/L) was acquired which verified the efficiency of the applied method.
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http://dx.doi.org/10.1016/j.bjm.2018.03.009DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6175716PMC
December 2018