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BACKGROUND: Butyrylcholineesterase (BChE) is a therapeutic drug and its producing as a recombinant protein is an essential issue in biotechnology. One of the highlights in this regard is choosing the best host cells and plasmids. OBJECTIVES: The aim of this study is to evaluate the production of butyrylcholinesterase in Vero, HEK-293, and CHO cell lines using a dual promoter vector. MATERIAL AND METHODS: The dual-promoter construction (pBudCE dual BChE) was transfected into cell lines categorized in three experimental groups (pBudCE dual BChE, pCMV and negative control). BChE gene expression and enzyme activity was evaluated at different times. RESULTS: All three cell lines showed higher gene expression level in pBudCE dual BChE group. BChE enzyme activity level of this group in CHO cells decreased in sixth day and increased in ninth day. In HEK-293 cells it has a downward trend from sixth to ninth day and in Vero cells its level in the ninth day was the highest. CONCLUSION: The difference of pBudCE dual BChE and pCMV groups was more pronounced in the HEK-293 cell and the BChE gene expression level of this cells was higher than the others while, CHO cells showed higher level of BChE enzyme activity.

Background: Chromene derivatives showed numerous biological activities. In the current study, the antioxidant, cytotoxicity, and apoptosis properties of halogenated dihydropyrano[3,2-b]chromene-3-carbonitrile derivatives (HDCCD) on MCF-7 cell line have been examined. Objectives: This study's principal point was synthesizing new halogenated pyranochromene derivatives and assessing their cytotoxic effects and apoptosis potential on MCF-7 breast cancer cell line by flow cytometry. Methods: Initially, 6-chloro- and 6-bromo-3-hydroxychromone compounds were prepared. In the next step, a series of HDCCD were synthesized by a one-pot three-component reaction of these two compounds, aromatic aldehydes, and malononitrile, in the presence of triethylamine in EtOH at reflux conditions. These compounds were fully characterized by standard spectroscopic techniques (IR, 1H, and 13C NMR) and elemental analyses. The potential of the antioxidant activity was determined by using ferric reducing antioxidant power assay (FRAP). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and lactate dehydrogenase (LDH) were used to evaluate metabolic activity. The nitric oxide (NO) and malondialdehyde (MDA) biomarkers of the exposed cells were evaluated on the cells and their supernatant. To quantify apoptotic death of MCF-7 breast cancer cells treated by the compounds at their IC50 concentrations, Annexin V-FITC apoptosis detection kit was utilized. Molecular docking of compounds (6a-j) into the Cyclin-dependent kinase 6 (PDB code: 4EZ5) was carried out, and the probable binding mode of compounds 6e and 6j was determined. Results: A dose-response relationship was seen in all the compounds. Most of them induced cytotoxic effects on the cells. Nitrite concentration of the culture media of the cells was decreased compared to the control. Malondialdehyde levels of the cells were below the range of the control by the addition of 6b, 6d, 6e, 6f, and 6g compounds on the cells, while the addition of the 6a, 6c, 6h, 6i, and 6j compounds increased the MDA level compared to the control. Flow cytometric analysis showed that most of the exposed cells were in the early and late apoptotic stage, and a few of them were in the necrotic stage. Conclusions: It could be concluded that HDCCD (6a-j) was toxic and caused death in the cells by apoptosis. The compounds have lipophilic characteristics, so they can easily pass the cell membrane. As confirmed by LDH results, it can be concluded that the cytotoxicity is connected with apoptosis rather than necrosis, endorsed by flowcytometry analysis afterward.

BACKGROUND: Candida albicans cause oral and vaginal mucosa infections as well as bloodstream and deep-tissue infections. Commonly, clotrimazole as a broad-spectrum antimitotic drug applied for treatment of Candida albicans infections. Bacterial ghosts are dead cells that have the broad potential to target the various body tissues and cells as drug vector. OBJECTIVES: We hope to conquest this resistance by using clotrimazole loaded on bacterial ghosts. METHODS: Lactobacillus ghosts were produced by using tween 80 and lactic acid according to the protocol and the amount of the DNA and protein in supernatant was measured by Nano-drop spectrophotometry. Ghost’s morphological characteristics were detected by using light microscopy, SEM and AFM. Antifungal activities of the synthesized ghosts were measured by plate methods. Three independent vertical Franz cells were used to evaluate drug release profile. BG-clotrimazole was added into cream base and was examined for dispensability as well as uniformity of the formulation on the skin. RESULTS: Results of the Nano-drop analysis showed that protein and DNA was seen in supernatant of treatment compared to control groups. AFM results showed well-dispersed and rod-shaped L. casei ghost cells. Lysis pores formation in the L. casei ghosts was indicated by SEM micrographs. BGs represent an excellent drug delivery system because of the high loading capability. Nearly, 50% of clotrimazole was released from BGs during 90 min. Highest anticandidal activity occurred using 100 mg/l clotrimazole-BG, while toxic effects were also seen with 10 mg/l clotrimazole. IC(50) clotrimazole-BG was found at 0.001 mg/l. Chemical stability results showed that about 90% of clotrimazole remained in the formulation. CONCLUSION: It could be concluded that the bacterial ghosts are very talented to high loading capability, keeping and releasing drug during six months, therefore these could act as an excellent drug delivery system. GRAPHIC ABSTRACT: [Image: see text]

Extracted cellulose fibers from Quinoa wastes were successfully incorporated with multi-walled carbon nanotubes/ZnO (MWCNTs/ZnO). Manganese complex was supported on natural cellulose and reacted with MWCNT/Zn bio-nanocomposite to have MWCNT/ZnO/Mn complex@oxidized cellulose. This novel bio-nanocomposite was characterized by SEM, TEM, FT-IR, TGA, and EDS. The catalytic activity of this composite was considered on the oxidation of alcohols. High conversion and selectivities of desired aldehyde and ketone products were obtained applying H2O2 and under solvent-free conditions. This heterogeneous catalyst was easily recycled and reused eight times without any significant catalytic activities and stabilities.

Cystic Echinococcosis (CE) is a parasitic infection caused by the larval stage of Echinococcus granulosus. Exploring safe and effective scolicidal agents for the surgery is an urgent need for the successful treatment of CE. This study aimed to determine scolicidal activity of the synthesized chitosan nanoparticles. Physicochemical properties of synthesized nanoparticles were determined by using DLS, FTIR, and SEM. Different concentrations of chitosan nanoparticles from 125 to 1000 μg/ml were examined at different incubation times (10, 60, 120, and 180 min). Scolicidal and cytotoxic activity of chitosan nanoparticles were confirmed by eosin exclusion and hemolysis activity tests. FTIR spectra, zeta potential (+42 ± 2.08) and PDI (0.388 ± 0.034) value revealed that the chitosan nanoparticles were synthesized. Significant differences among the scolicidal effects of chitosan nanoparticles were observed in comparison to the control treatments and highest scolicidal activity was observed at 1000 μg/ml after 180 min exposure time. Hemolytic activity was not significant at all concentrations of chitosan nanoparticles. Our findings support the hypothesis that Chitosan nanoparticles have the potential to be a safe and efficient scolicidal agent candidate at very low concentrations and in a wide range of exposure time. Further in vivo studies are recommended to evaluate chitosan nanoparticle efficacy before clinical application.

Context: One of the most common aggressive and primary brain tumors is glioma. The majority of diagnoses are referred to high-grade malignant glioblastoma, which carries the worst prognosis. Still, treatment of brain tumors remains a big challenge for clinicians. This study was designed to investigate the efficacy of gene therapy in the treatment of brain cancer. Methods: Studies use genes as a therapeutic agent in brain cancer treatment even alone or in combination with other treatment methods. Full-text papers, which met the inclusion criteria, were independently assessed by two reviewers. Disagreements were resolved by consultation with a third reviewer. Results: Statistical analysis showed that 50% of the papers used a virus, 36% used polymers, and 14% used cells as carriers to transfect the genes as a therapeutic agent in brain tumor models. Data showed that the estimated size of the brain tumor was reduced by using co-treatment of the gene with one of the conventional therapies. Conclusions: According to the results, co-treatment of the gene with conventional therapies could be more effective than the monotherapy methods.

In the present work, the lipopeptide biosurfactant (LPB) produced by Acinetobacter junii B6 was applied for the production of gold nanoparticles (Au NPs). Several methods, including transmission electron microscopy (TEM), UV-visible spectroscopy, Fourier transform infrared spectroscopy (FTIR), dynamic light scattering (DLS), and scanning electron microscopy (SEM), were used to determine the properties of the synthesized Au NPs. TEM and SEM images of Au NPs showed spherical and mostly hexagon-shaped and well-dispersed nanoparticles in the range of 10–17 nm. Cytotoxic activity of the Au NPs (assisted by WST-1 assay) revealed IC50 (μg/ml) of 89.08 ± 0.4, 646.12 ± 0.5, 3.37 ± 0.1, and 770 ± 0.15 against cell lines of U87, A549, MCF7, and 3T3, respectively. However, the anti-proliferative activities of biogenic Au NPs were significantly lower (IC50 of 770 ± 0.15 μg/mL) than HAuCl4 (IC50 of 0.26 ± 0.03 μg/mL) against 3T3 normal cell. The attained results of the antibacterial effect of the produced Au NPs and Au3+ ions revealed that Au3+ ions had a higher inhibitory effect than Au NPs against 3 Gram-positive and 4 Gram-negative bacterial strains. In the present study, we introduced a green approach for the synthesis of Au NPs using non-toxic and biodegradable LPBs in the absence of any chemical reducing agent.

Purpose The main aim of this review was to provide an updated comprehensive report regarding isolation methods of MSCs from human extra embryonic tissues, including cord blood, amniotic fluid, and different parts of the placenta and umbilical cord, with respect to the efficacy of these methods. Results Extra embryonic tissues are the most available source for harvesting of mesenchymal stem cells (MSCs). They make a large number of cells accessible using non-invasive methods of isolation and the least immune-rejection reactions. A successful culture of primary cells requires obtaining a maximum yield of functional and viable cells from the tissues. In addition, there are many reports associated with their differentiation into various kinds of cells, and there are some clinical trials regarding their utilization for patients. Conclusion Currently, cord blood-MSCs have been tested for cartilage and lung diseases. Umbilical cord-MSCs were tested for liver and neural disorders. However, these MSCs can be isolated, expanded, and cryopreserved in a cell bank for patients in need.

Purpose Genetic diseases can be the result of genetic dysfunctions that happen due to some inhibitory and/or environmental risk factors, which are mostly called mutations. One of the most promising treatments for these diseases is correcting the faulty gene. Gene delivery systems are an important issue in improving the gene therapy efficiency. Therefore, the main purpose of this study was modifying graphene oxide nanoparticles by spermine in order to optimize the gene delivery system. Methods Graphene oxide/APTES was modified by spermine (GOAS) and characterized by FT-IR, DLS, SEM and AFM techniques. Then pEGFP-p53 was loaded on GOAS, transfected into cells and evaluated by fluorescent microscopy and gene expression techniques. Results FT-IR data approved the GOAS sheet formation. Ninety percent of the particles were less than 56 nm based on DLS analysis. SEM analysis indicated that the sheets were dispersed with no aggregation. AFM results confirmed the dispersed structures with thickness of 1.25±0.87 nm. STA analysis showed that GOAS started to decompose from 400°C and was very unstable during the heating process. The first weight loss up to 200°C was due to the evaporation of absorbed water, the second one observed in the range of 200-550°C was assigned to the decomposition of labile oxygen- and nitrogen-containing functional groups, and the third one above 550°C was attributed to the removal of oxygen functionalities. In vitro release of DNA demonstrated the efficient activity of the new synthesized system. Ninety percent of the cells were transfected and showed the GFP under fluorescence microscopy, and TP53 gene was expressed 51-fold in BT-20 cells compared to β-actin as the reference gene. Flow cytometry analysis confirmed the apoptosis of the cells rather than necrosis. Conclusion It could be concluded that the new synthesized structure could transfer a high amount of the therapeutic agent into cells with best activity.

Background Microemulsions drug delivery systems (MDDS) have been known to increase the bioavailability of hydrophobic drugs. The main challenge of the MDDS is the development of an effective and safe system for drug carriage and delivery. Biosurfactants are preferred surface-active molecules because of their lower toxicity and safe characteristics when compared to synthetic surfactants. Glycolipid and lipopeptide are the most common biosurfactants that were tested for MDDS. The main goal of the present systematic review was to estimate the available evidence on the role of biosurfactant in the development of MDDS. Search strategy Literature searches involved the main scientific databases and were focused on the period from 2005 until 2017. The Search filter composed of two items: "Biosurfactant" and/or "Microemulsion." Inclusion criteria Twenty-four studies evaluating the use of biosurfactant in MDDS were eligible for inclusion. Among these 14 were related to the use of glycolipid biosurfactants in the MDDS formulations, while four reported using lipopeptide biosurfactants and six other related review articles. Results According to the output study parameters, biosurfactants acted as active stabilizers, hydrophilic or hydrophobic linkers and safety carriers in MDDS, and among them glycolipid biosurfactants had the most application in MDDS formulations. Conclusion Synthetic surfactants could be replaced by biosurfactants as an effective bio-source for MDDS due to their excellent self-assembling and emulsifying activity properties.

Purpose: The main aim of the present study was to design, fabrication and physicochemicalcharacteristics of the magnetogel nanospheres as carriers for Cisplatin in the in vitro environment.Methods: Magnetic nanospheres were synthesized by using a chemical co-precipitation methodand coated by sodium alginate through double emulsion method. Then cisplatin was encapsulatedinto β-cyclodextrin -sodium alginate grafted magnetic nanospheres. The physicochemicalproperties of the sodium alginate grafted magnetic nanospheres were characterized by using FTIR,particle size analyzing, vibrating sample magnetometry, thermogravimetric and SEM analysis.Also the drug entrapment efficiency, content and in vitro release profile were investigated.Results: Size distribution results revealed that the particles size was distributed in the rangeof 50± nm. Also morphological properties showed that particles are separated and sphericalwith the grafted layers of the polymer. The release profile data were in the acceptable rangecompared to the blank (cisplatin solution).Conclusion: It could be concluded that the sodium alginate grafted magnetic nanospheres couldact as a slow and controlled release system to deliver cisplatin.

The main aim of this study was to synthesize the superparamagnetic nanoparticles coated by alginate/chitosan/β-cyclodextrin to purify α-amylase. Isolated bacteria were identified by morphological, biochemical and taxonomic molecular studies. FTIR- spectrometer, VSM, X-ray instruments and Malvern Zetasizer were used to characterize nanoparticles characteristics. The morphological structures and the elemental composition of the nanoparticles were studied by using FESEM and EDS, respectively. The molecular weight of enzyme was determined using SDS-PAGE, and the enzyme activity detected by zymographic analysis. FTIR studies showed the presence of Fe–O–Fe in the Fe3O4 and verified the interaction between chitosan, β-cyclodextrin and alginate. The saturation magnetization for superparamagnetic and coated superparamagnetic nanoparticles was indicated 39 and 1.9 emu g−1, respectively. The maximum intensity of the XRD peak indicated the presence of the Fe3O4. FESEM and EDS analysis showed that the nanopart...

Background Magnetic nanoparticles (MNPs) loaded by various active compounds can be used for targeted drug delivery. Objectives: In the present study, the Fe3O4 magnetic nanoparticles that contained gentamicin were prepared and their antibacterial activities were studied. Materials and Methods MNPs containing gentamicin (G@SA-MNPs) were prepared using sodium alginate (SA) as a surface modifier. After and before coating, the prepared MNPs were characterized using transmission electron microscopy (TEM), X-ray diffraction spectroscopy (XRD), Fourier transform infrared spectroscopy (FTIR), and vibrating sample magnetometer (VSM). Finally, the antibacterial effect of the MNPs was investigated by a conventional serial agar dilution method. Results Particle size distribution analysis showed that the size of MNPs, before and after coating, was in the range of 1–18 nm and 12–40 nm, respectively. The magnetization curve of G@SA-MNPs (with saturation magnetization of 27.9 emu.g-1) confirmed ferromagnetic property. Loading gentamicin on the surface of MNPs was qualitatively verified by FTIR spectrum. Quantitative analysis measurements indicated the gentamicin loading on SA-MNPs as 56.7 ± 5.4%. The measured MICs of G@SA-MNPs for Pseudomonas aeruginosa (PTTC 1574) was 1.28 µg.mL-1. The sub-MIC (0.64 µg.mL-1) concentration of G@SA-MNPs in nutrient broth could successfully inhibit the growth of P. aeruginosa for 14 hours. Conclusions Loading gentamicin on the SA-MNPs exhibited reasonable antibacterial effects against P. aeruginosa.

Elham Afzali,1 Touba Eslaminejad,1 Seyede Elmira Yazdi Rouholamini,2 Mariam Shahrokhi-Farjah,2 Mehdi Ansari3 1Pharmaceutics Research Centre, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran; 2Physiology Research Centre, Institute of Basic and Clinical Physiology Sciences, Kerman University of Medical Sciences, Kerman, Iran; 3Department of Drug and Food Control, Faculty of Pharmacy, Kerman University of Medical Sciences, Kerman, IranCorrespondence: Mariam Shahrokhi-Farjah; Touba Eslaminejad Tel +983431325243Fax +983431325003Email M_Shahrokhi@KMU.ac.ir; t_eslami@kmu.ac.irPurpose: Breast cancer is one of the most lethal types of cancer in women. Curcumin showed therapeutic potential against breast cancer, but applying that by itself does not lead to the associated health benefits due to its poor bioavailability, which appears to be primarily due to poor absorption, rapid metabolism, and rapid elimination. Moreover, poor water solubility of curcumin causes accumulation of a high concentration of curcumin and so decrease its permeability to the cell. Many strategies are employed to reduce curcumin metabolism such as adjuvants and designing novel delivery systems. Therefore, in this study sodium alginate and chitosan were used to synthesize the hydrogels that are known as biocompatible, hydrophilic and low toxic drug delivery systems. Also, folic acid was used to link to chitosan in order to actively targetfolate receptors on the cells.Methods: Chitosan-β-cyclodextrin-TPP-Folic acid/alginate nanoparticles were synthesized and then curcumin was loaded on them. Interaction between the constituents of the particles was characterized by FTIR spectroscopy. Morphological structures of samples were studied by FE-SEM. Release profile of curcumin was determined by dialysis membrane. The cytotoxic test was done on the Kerman male breast cancer (KMBC-10) cell line by using MTT assay. The viability of cells was detected by fluorescent staining. Gene expression was investigated by real-time PCR.Results: The encapsulation of curcumin into nano-particles showed an almost spherical shape and an average particle size of 155 nm. In vitro cytotoxicity investigation was indicated as dose-respond reaction against cancer breast cells after 24 h incubation. On the other hand, in vitro cell uptake study revealed active targeting of CUR-NPs into spheroids. Besides, CXCR4 expression was detected about 30-fold less than curcumin alone. The CUR-NPs inhibited proliferation and increased apoptosis in spheroid human breast cancer cells.Conclusion: Our results showed the potential of NPs as an effective candidate for curcumin delivery to the target tumor spheroids that confirmed the creatable role of folate receptors.Keywords: spheroids cell, curcumin, nanospheres, cytotoxicity, chitosan, alginate

It is about Crona virus as a pandemic disease.

Purpose Genetic diseases can be the result of genetic dysfunctions that happen due to some inhibitory and/or environmental risk factors, which are mostly called mutations. One of the most promising treatments for these diseases is correcting the faulty gene. Gene delivery systems are an important issue in improving the gene therapy efficiency. Therefore, the main purpose of this study was modifying graphene oxide nanoparticles by spermine in order to optimize the gene delivery system. Methods Graphene oxide/APTES was modified by spermine (GOAS) and characterized by FT-IR, DLS, SEM and AFM techniques. Then pEGFP-p53 was loaded on GOAS, transfected into cells and evaluated by fluorescent microscopy and gene expression techniques. Results FT-IR data approved the GOAS sheet formation. Ninety percent of the particles were less than 56 nm based on DLS analysis. SEM analysis indicated that the sheets were dispersed with no aggregation. AFM results confirmed the dispersed structures with thickness of 1.25±0.87 nm. STA analysis showed that GOAS started to decompose from 400°C and was very unstable during the heating process. The first weight loss up to 200°C was due to the evaporation of absorbed water, the second one observed in the range of 200–550°C was assigned to the decomposition of labile oxygen- and nitrogen-containing functional groups, and the third one above 550°C was attributed to the removal of oxygen functionalities. In vitro release of DNA demonstrated the efficient activity of the new synthesized system. Ninety percent of the cells were transfected and showed the GFP under fluorescence microscopy, and TP53 gene was expressed 51-fold in BT-20 cells compared to β-actin as the reference gene. Flow cytometry analysis confirmed the apoptosis of the cells rather than necrosis. Conclusion It could be concluded that the new synthesized structure could transfer a high amount of the therapeutic agent into cells with best activity.

Purpose The main aim of this review was to provide an updated comprehensive report regarding isolation methods of MSCs from human extra embryonic tissues, including cord blood, amniotic fluid, and different parts of the placenta and umbilical cord, with respect to the efficacy of these methods. Results Extra embryonic tissues are the most available source for harvesting of mesenchymal stem cells (MSCs). They make a large number of cells accessible using non-invasive methods of isolation and the least immune-rejection reactions. A successful culture of primary cells requires obtaining a maximum yield of functional and viable cells from the tissues. In addition, there are many reports associated with their differentiation into various kinds of cells, and there are some clinical trials regarding their utilization for patients. Conclusion Currently, cord blood-MSCs have been tested for cartilage and lung diseases. Umbilical cord-MSCs were tested for liver and neural disorders. However, these MSCs can be isolated, expanded, and cryopreserved in a cell bank for patients in need.

Background Recently, the development of a safe bioactive material with antioxidant properties, which can improve healing activity are focusing. Biosurfactants are very famous for their antimicrobial and free radical scavenging activities. Thereof, the main aim of the present study was to investigate the antioxidant and wound healing activity of the lipopeptide biosurfactant (LBS) produced by Acinetobacter junii B6. DPPH radical scavenging activities and FRAP assays were used to measure the antioxidant properties. Methods For evaluation of the wound healing activity, 36 rats (previously wounded in depilated thoracic region) were randomly distributed into six groups and chromatic, wound contraction, and histopathological feature were examined. The assessment levels of reactive oxygen species (ROS) after LBS exposure were determined using malondialdehyde (MDA), hydrogen peroxide (H2O2), and glutathione (GSH) assay kits. Results DPPH assay showed notable scavenging activities at the corresponding concentrations with IC50 value of 0.7 mg/ml. The reductive potency of the LBS showed lower performance at low concentration, while exhibited a remarkable increase at higher concentration. The best histopathological remission was achieved following treatment by 5 mg/ml of the LBS. Scar wounds at day 13 showed the lowest lesion sizes, increased re-epithelialization, hair follicle detection, and decreased amounts of neutrophilic inflammation, immaturity of the wound bed, erythema, edema, capillary, and retention of necrotic tissue. Conclusion Results from MDA, H2O2, and GSH levels of the treated sample confirmed the scavenging property of the bacterial derived LBS through ROS. It could be concluded that the pharmaceutical formula encourages the wound healing because of its notable antioxidant capacity. Highlights • DPPH and FRAP assays showed notable scavenging activity. • MDA, H2O2, and GSH; confirmed the scavenging property of the derived biosurfactant through ROS. • Synthesized formula encourages the healing of the wound because of its antioxidant capacity.

Background Microemulsions drug delivery systems (MDDS) have been known to increase the bioavailability of hydrophobic drugs. The main challenge of the MDDS is the development of an effective and safe system for drug carriage and delivery. Biosurfactants are preferred surface-active molecules because of their lower toxicity and safe characteristics when compared to synthetic surfactants. Glycolipid and lipopeptide are the most common biosurfactants that were tested for MDDS. The main goal of the present systematic review was to estimate the available evidence on the role of biosurfactant in the development of MDDS. Search Strategy Literature searches involved the main scientific databases and were focused on the period from 2005 until 2017. The Search filter composed of two items: “Biosurfactant” and/or “Microemulsion.” Inclusion Criteria Twenty-four studies evaluating the use of biosurfactant in MDDS were eligible for inclusion. Among these 14 were related to the use of glycolipid biosurfactants in the MDDS formulations, while four reported using lipopeptide biosurfactants and six other related review articles. Results According to the output study parameters, biosurfactants acted as active stabilizers, hydrophilic or hydrophobic linkers and safety carriers in MDDS, and among them glycolipid biosurfactants had the most application in MDDS formulations. Conclusion Synthetic surfactants could be replaced by biosurfactants as an effective bio-source for MDDS due to their excellent self-assembling and emulsifying activity properties.

Magnetic nanoparticles have been used as effective vehicles for the targeted delivery of therapeutic agents that can be controlled in their concentration and distribution to a desired part of the body by using externally driven magnets. This study focuses on the synthesis, characterization, and functionalization of pullulan–spermine (PS) magnetic nanoparticles for medical applications. Magnetite nanopowder was produced by thermal decomposition of goethite (FeOOH) in oleic acid and 1-octadecene; pullulan–spermine was deposited on the magnetite nanoparticles in the form of pullulan–spermine clusters. EGFP-p53 plasmid was loaded on functionalized iron oleate to transfer into cells. Synthesized nanoparticles were characterized by Fourier transform infrared spectroscopy (FTIR), dynamic light scattering (DLS), vibrating sample magnetometry (VSM), and transmission electron microscopy (TEM). The encapsulation efficiency and drug loading efficiency of the nanocomplexes were tested. FTIR studies showed the presence of oleic acid and 1-octadecene in the iron oleate nanopowder and verified the interaction between spermine and pullulan. The characteristic bands of PS in the spectrum of the pullulan–spermine-coated iron oleate (PSCFO) confirmed that PS covered the surface of the iron oleate particles. TEM studies showed the average size of the iron oleate nanopowder, the PSCFO, and the plasmid-carrying PSCFO (PSCFO/pEGFP-p53) to be 34±12 nm, 100±50 nm and 172±3 nm, respectively. Magnetic measurements revealed that magnetic saturation of the PSCFO was lower in comparison with the iron oleate nanopowder due to the presence of organic compounds in the former. In cytotoxicity tests performed using U87 cells as glioblastoma cells, a 92% survival rate was observed at 50 µg/µl of the plasmid-carrying PSCFO, with an IC50 value of 189 µg/µl.

Aims: Applications of herbal medicine extracts as antimicrobials agents in human health and food protection as well as the analysis methods of the novel compounds, biosynthesis, and genetic regulation of them are describing in this work. According to the increasing prevalence of the antibiotic-resistant microorganisms, the alternative source of antibiotic production is necessary to replace. Plants are a valuable source in various secondary metabolites such as alkaloids, flavonoids, tannins, and terpenoids which have shown some inhibitory effect on different microorganisms. Nowadays, herbal medicines are finding their way into pharmaceuticals. There are several well-known drugs that are derived from plants used for different diseases and were topselling pharmaceuticals in the world. ‘Thymol’ is a strong disinfectant from ‘Thyme’ for treating of edema and bronchospasm, ‘morphine’ which comes from ‘Papaver somniferum’, the powerful painkiller, ‘L-dopa’ from ‘Mucuna deeringiana’ using as a cardiotonic, ‘taxol’ from the bark of ‘Taxus Review Article Sharififar et al.; EJMP, 11(1): 1-17, 2016; Article no.EJMP.20114 2 brevifolia’ as an anti-cancerous and ‘Rutin’ from ‘Orange’ using to treat sedation. In conclusion, plant medicine extracts have used in ancient medicine from several years ago, so identification of them and their compounds offer a chance to develop new drugs against diseases and according to this potential, need further work to approve reliability. Study Design and Methodology: A review of the literature from the pioneering study of 2005 until 2015. Place and Duration of Study: Pharmaceutics Research Centre (PRC), Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran between June 2014 and July 2015. Conclusion: Plant medicine extracts have used in ancient medicine from several years ago, so identification of them and their compounds offer a chance to develop new drugs against diseases and according to this potential, need further work to approve reliability.

Lipopeptide biosurfactants (LPBs) are amphiphilic compounds produced by microorganisms exhibiting various biological activities. The main aim of the present study was to assess the in vitro antimicrobial, anti-biofilm, and cytotoxic effects of LPB produced by Acinetobacter junii (AjL). We determined AjL minimum inhibitory concentration (MIC) against both Gram-positive and Gram-negative bacteria as well as two fungal strains. Also, the anti-biofilm activity of AjL against the biofilm produced by clinically isolated bacterial strains was investigated. The AjL non-selectively showed activity against both Gram-positive and Gram-negative bacterial strains. The obtained results of the present study exhibited that the AjL in concentrations nearly below critical micelle concentration (CMC) has an effective antibacterial activity. It was found that the MIC values of AjL were lower than standard antifungal and it exhibited nearly 100% inhibition against Candida utilis. The attained results of the biofilm formation revealed that AjL disrupted the biofilm of Proteus mirabilis, Staphylococcus aureus, and Pseudomonas aeruginosa at 1250 μg/ml and 2500 μg/ml concentrations. The attained results of cytotoxic effect (determined by WST-1 assay) of the AjL revealed IC50 of 7.8 ± 0.4 mg/ml, 2.4 ± 0.5 mg/ml, and 5.7 ± 0.1 mg/ml, against U87, KB, and HUVEC cell lines, respectively. The results indicated that AjL has a potential application in the relatively new field of biomedicine.

Differentiation of mesenchymal stem cells (MSCs) into cardiomyocytes is a complex phenomenon, and attempts to find an effective inducing agent are still ongoing. We studied the effect of fibrin scaffold and sodium valproate (VPA, as a histone deacetylase inhibitor) on the differentiation of human adipose-derived stem cells (hADSCs) into cardiomyocyte-like cells. The cells were cultured in culture flask (2D) and in fibrin scaffold (3D), fabricated of human plasma fibrinogen, with and without VPA (1 mM). QRT-PCR, Western blot, and immunochemistry assays were used to evaluate the expression of cardiac markers at gene and protein levels. High levels of CD44, CD90, CD73, and CD105 were expressed on the surface of hADSCs. Treated encapsulated hADSCs (3D) presented significantly higher mRNA expression of HAND1 (1.54-fold), HAND2 (1.59-fold), cTnI (1.76-fold), MLC2v (1.4-fold), Cx43 (1.38-fold), βMHC (1.34-fold), GATA4 (1.48-fold), and NKX2.5 (1.66-fold) in comparison to 2D conditions at four weeks after induction. The protein expressions of NKX2.5 (0.78 vs 0.65), cTnI (1.04 vs 0.81), and Cx43 (1.11 vs 1.08) were observed in the differentiated cells both in 3D and 2D groups, while control cells were absolutely negative for these proteins. The frequency of cTnI and Cx43-positive cells was significantly higher in 3D (24.2 ± 15 and 12 ± 3%) than 2D conditions (19.8 ± 3 and 10 ± 2%). Overall, the results showed that VPA can increase cardiomyogenesis in hADSCs and that fibrin scaffold enhances the inductive effect of VPA. Results of this study may improve cell-based protocols for implementation of more successful cardiac repair strategies.

Commonly, acquired resistances to anticancer drug are mediated by overexpression of a membrane-associated protein that encode via multi-drug resistance gene-1 (MDR1). Herein, the mRNA-cleaving DNAzyme that targets the mRNA of MDR1 gene in doxorubicin-resistant breast cancer cell line (MCF-7/DR) loaded on the chitosan β-cyclodextrin complexes was used as a tropical agent. Chitosan/β-cyclodextrin complexes were used to deliver DNAzymes into cancer cells. Determination of the physicochemical characteristics of the particles was done by photon correlation spectroscopy and scanning electron microscopy. The encapsulation efficiency of the complexes was tested by using gel retardation assay. Positively charged nanoparticles interacted with DNAzyme that could perform as an efficient DNAzyme transfection system. The rationale usage of this platform is to sensitize MCF-7/DR to doxorubicin by downregulating the drug-resistance gene MDR1. Results demonstrated a downregulation of MDR1 mRNAs in MCF-7/DR/DNZ by real-time PCR, compared to the MCF-7/DR as control. WST1 assay showed the 22-fold decrease in drug resistance on treated cells 24 h after transfection. Results showed the intracellular accumulation of Rh123 increased in the treated cells with DNAzyme. Results suggested a potential platform in association with chemotherapy drug for cancer therapy and indicated extremely efficient at delivery of DNAzyme in restoring chemosensitivity.

Background Blood-brain barrier (BBB) separates the neural tissue from circulating blood because of its high selectivity. This study focused on the in vitro application of magnetic nanoparticles to deliver Tp53 as a gene of interest to glioblastoma (U87) cells across a simulated BBB model that comprised KB cells. Material and method After magnetic and non-magnetic nanoparticles were internalized by KB cells, their location in these cells was examined by transmission electron microscopy. Transfection efficiency of DNA to U87 cells was evaluated by fluorescence microscopy, real time PCR, flowcytometry, and Western immuno-blotting. When a magnetic field was applied, a large number of magnetic nanoparticles accumulated in KB cells, appearing as black dots scattered in the cytoplasm of cells. Fluorescence microscope examination showed that transfection of the DNA to U87 target cells was highest in cells treated with magnetic nanoparticles and exposed to a magnetic field. Also it was reflected in significantly increased mRNA level while the p53 protein level was decreased. Conclusion It could be concluded that a significant increase in total apoptosis was induced in cells by magnetic nanoparticles, coupled with exposure to a magnetic force (p ≤0.01) as compared with cells that were not exposed to magnetism.

In this study cationic β-cyclodextrin-chitosan-mediated nanoparticles were used to transfer pmCherry-C1 into glioblastoma cells and their transfection efficiency were compared to lipofectamine and electroporation. Physicochemical characteristics of nanoparticles were evaluated by photon correlation spectroscopy and scanning electron microscopy. Electrophoretic nuclease resistance and stability assays were used to check the protection of DNA from nucleases digestion. mCherry reporter construct was used for visualization, followed by quantitation of cell survival and gene expression by fluorescence-activated cell sorting analysis and fluorescence microscopy. Particle size was approximately 200 nm and did not change at 4 °C even after 12 weeks. Importantly, the positively charged complexes interacted with DNA could serve as an efficient DNA delivery systems. Most of the gene was associated with the nanoparticles and was efficiently protected from DNAse I digestion. More than 80 % of transfected cells expressed mCherry efficiently.

Background Ischemic stroke is an acute brain insult that induces dramatic changes in the neurons. Treatment of brain stroke is one of the main therapeutic targets of neuroprotective therapies. The aim of this study was to evaluate the protective potential of implanted human umbilical cord mesenchymal stem (hUCMs) cells with/without aspirin (ASA) against focal cerebral ischemia. Methods We assessed the migration and distribution of PKH 26 -labeled cells after transplantation. After day 10 of transient occlusion, we evaluated the effect of ASA and hUCMs on the recovery of learning and memory in rats by Morris water maze. Afterward, animals were sacrificed, and the infarct area in the brain was evaluated using 2, 3, 5-triphenyltetrazolium chloride staining and also by hematoxylin and eosin. Results The recovery of learning and memory in ischemic animals that received ASA and hUCM cells improved significantly compared with the untreated ischemic animals. Coadministration of ASA and hUCM cells did not improve the outcome at a comparable rate with ASA and hUCM cells alone. PKH 26 -labeled cells were detectable in the ischemic area of the brain tissue sections. 2,3,5-Triphenyltetrazolium chloride staining and histologic examinations showed that treatment with ASA and hUCM cells could significantly alter the ischemic area. Conclusions The results of the present study suggest that ASA and hUCM cells can withstand degenerative changes induced by artificial stroke in the rat. Also the learning and memory disturbance in the ASA and cell-treated animals is less pronounced than ischemic animals. Coadministration of ASA and hUCM cells did not raise the outcome higher than administration of ASA and hUCM cells alone.

The Publisher regrets that this article is an accidental duplication of an article that has already been published, doi:10.1016/j.micpath.2012.05.010;. The duplicate article has therefore been withdrawn.

Roselle, or Jamaica sorrel (Hibiscus sabdariffa) is a popular vegetable in many tropical regions, cultivated for its leaves, seeds, stems and calyces which, the dried calyces are used to prepare tea, syrup, jams and jellies and as beverages. The main objectives of this study were to identify and characterise fungal pathogens associated with Roselle diseases based on their morphological and cultural characteristics and to determine the pathogenicity of four fungi infecting Roselle seedlings, namely Phoma exigua, Fusarium nygamai, Fusarium tgcq and Rhizoctonia solani in Penang. A total of 200 fungal isolates were obtained from 90 samples of symptomatic Roselle tissues. The isolates were identified based on cultural and morphological characteristics, as well as their pathogenicity. The fungal pathogen most frequently isolated was P. exigua (present in 45% of the samples), followed by F. nygamai (25%), Rhizoctonia solani (19%) and F. camptoceras (11%). Pathogenicity tests showed that P. exigua, F. nygamai, F. camptoceras and R. solani were able to infect both wounded and unwounded seedlings with different degrees of severity as indicated by the Disease severity (DS). R. solani was the most pathogenic fungus affecting both wounded and unwounded Roselle seedlings, followed by P. exigua that was highly pathogenic on wounded seedlings. F. nygamai was less pathogenic while the least pathogenic fungus was F. camptoceras, infecting only the unwounded seedlings but, surprisingly, not the wounded plants.