Isolation and identification of endophytic actinobacteria from Iris persica and Echium amoenum plants and investigation of their effects on germination and growth of wheat plant.

Plant biotechnology helps to develop different types of new products with increased resistance to disease, greater tolerance to drought and salt stress, and better nutritional value. The interaction of plants and microorganisms will play a significant role to achieve this purpose. The aims of this study were to isolate endophyte Actinobacteria strains of some medicinal plants and the investigation of their bioactive potential. 15 Actinobacteria strains were selectively isolated from Persian iris and Echium amoenum plants, and then their belonging to Actinobacteria phylum was confirmed using an Actinobacteria-specific primer pair. The antioxidant activity of the crude extract obtained from the isolated strains was investigated based on DPPH method. Investigating the antioxidant activity of the crude extract showed that at a concentration of 100 µg/mL, the two strains EG1 and EG2 had 71% and 78% antioxidant activity, respectively. According to the phylogeny studies, it was determined that two strains belonged to the Streptomyces genus. The effect of supernatant achieved from selected endophytic strain on 35-day wheat plants showed that the supernatant considerably promotes root and shoot growth and chlorophyll content under salinity stress (150 mM NaCl). In general, it can be concluded strains that live symbiotically with medicinal plants are rich sources of bioactive compounds. Therefore, identification of the bioactive compounds in the extract of isolated Actinobacteria from medicinal plants and further studies on their metabolism are suggested.

Pediococcus acidilactici isolated from traditional cheese as a potential probiotic with cytotoxic activity against doxorubicin-resistant MCF-7 cells.

The considerable flexibility of cancerous cells to escape from chemical and biological drugs makes it clear that much is to be done to control and eliminate such cells. Probiotic bacteria, in this regard, have shown promising performance. In this study, we isolated and characterized lactic acid bacteria from traditional cheese. Then we evaluated their activity against doxorubicin-resistant MCF-7 cells (MCF-7/DOX) through MTT assay, Annexin V/PI protocol, real-time PCR, and western blotting. Among the isolates, one strain with more than 97% similarity with Pediococcus acidilactici showed considerable probiotics properties. Low pH, high bile salts, and NaCl could not significantly affect this strain while it was susceptible to antibiotics. Also, it had a potent antibacterial activity. Besides, the cell-free supernatant of this strain (CFS) significantly reduced the viability of MCF-7 and MCF-7/DOX cancerous cells (to about 10% and 25%, respectively), while it was safe for normal cells. Also, we found that CFS could regulate the Bax/Bcl-2 at mRNA and protein levels to induce apoptosis in drug-resistant cells. We determined 75% early apoptosis, 10% late apoptosis, and 15% necrosis in the cells treated with the CFS. These findings can accelerate the development of probiotics as promising alternative treatments to overcome drug-resistant cancers.

Penicillin and Oxacillin Loaded on PEGylated-Graphene Oxide to Enhance the Activity of the Antibiotics against Methicillin-Resistant Staphylococcus aureus.

Infectious diseases are known as the second biggest cause of death worldwide, due to the development of antibiotic resistance. To overcome this problem, nanotechnology offers some promising approaches, such as drug delivery systems that can enhance drug efficiency. Herein, a Graphene Oxide-polyethylene glycol (GO-PEG) nano-platform was synthesized and penicillin and oxacillin, two antibiotics that are ineffective against Methicillin-resistant S. aureus (MRSA), were loaded on it to improve their effectiveness. The nanocomposites were characterized using FTIR, XRD, UV-Vis, FE-SEM/EDX, and Zeta potential analyses, followed by an evaluation of their antibacterial activity toward MRSA. Based on the results, drug loaded GO-PEG nanocomposites with loading efficiencies of 81% and 92% for penicillin and oxacillin, respectively, were successfully synthesized. They showed a controlled release within six days. The zeta potential of GO-PEG-oxacillin and penicillin was -13 mV and -11 mV, respectively. The composites showed much more activity against MRSA (80-85% inhibition) in comparison to GO-PEG (almost 0% inhibition) and pure antibiotics (40-45% inhibition). SEM images of MRSA treated with GO-PEG-antibiotics showed a deformation in the structure of bacterial cells, which led to the collapse of their intracellular components. These results demonstrate the effectiveness of utilizing the GO-based nanoplatforms in enhancing the antibacterial activity of the antibiotics.

Down-regulation of biofilm-associated genes in mecA-positive methicillin-resistant S. aureus treated with M. communis extract and its antibacterial activity.

Considering the prevalence of resistance to antibiotics, the discovery of effective agents against resistant pathogens is of extreme urgency. Herein, 26 mecA-positive methicillin-resistant S. aureus (MRSA) isolated from clinical samples were identified, and their resistance to 11 antibiotics was investigated. Next, the antibacterial and anti-biofilm activity of the ethanolic extract of M. communis on these strains was evaluated. Furthermore, the effect of this extract on the expression of biofilm-associated genes, icaA, icaD, bap, sarA, and agr, was studied. According to the results, all isolated strains were multidrug-resistant and showed resistance to oxacillin and tetracycline. Also, 96.15 and 88.46 % of them were resistant to gentamicin and erythromycin. However, the extract could effectively combat the strains. The minimum inhibitory concentration (MIC) against different strains ranged from 1.56 to 25 mg/ml and the minimum bactericidal concentration (MBC) was between 3.125 and 50 mg/ml. Even though most MRSA (67 %) strongly produced biofilm, the sub-MIC concentration of the extract destroyed the pre-formed biofilm and affected the bacterial cells inside the biofilm. It could also inhibit biofilm development by significantly decreasing the expression of icaA, icaD, sarA and bap genes involved in biofilm formation and development. In conclusion, the extract inhibits biofilm formation, ruins pre-formed biofilm, and kills cells living inside the biofilm. Furthermore, it down-regulates the expression of necessary genes and nips the biofilm formation in the bud.

Probiotic potential and anticancer properties of Pediococcus sp. isolated from traditional dairy products.

Herein, 18 lactic acid bacteria isolated from 30 samples of traditional dairy products were identified, and their probiotic potential was evaluated. According to the results, almost all strains showed the probiotic properties sufficiently, though M1 had better characterise. 16S rRNA gene sequencing revealed that this strain belongs to the Pediococcus sp. (<95 % similarity). This strain had substantial antipathogenic activity and did not show any worrying antibiotic resistance. Also, the strain was resistant to high concentrations of bile salt (1 %), NaCl (6.5 %), and low pH (2). Furthermore, it was revealed that cell-free supernatant (CFS), heat-killed cells and live cells derived from M1 significantly decreased the viability of MCF-7 cells so that the CFS resulted in 85 % cell death. Flow cytometry and western blot analysis determined that this compound induced apoptosis in the cancerous cells through increasing the BAX protein expression and decreasing the Bcl-2 protein expression.

Bacteriostatic activity and partial characterization of the bacteriocin produced by L. plantarum sp. isolated from traditional sourdough.

This study was aimed to isolate and partially characterizes the bacteriocin produced by an L. plantarum sp. isolated from traditional sourdough. The bacteriocin was partially purified, and after treating it with different harsh conditions, its antibacterial activity was evaluated against L. monocytogenes as an indicator. Also, the growth phase during which the bacteriocin is produced, and its mode of action, was examined. Finally, the molecular weight of this compound was evaluated by using SDS-PAGE analysis. According to the results, this bacteriocin had a molecular weight well lower than 10 kDa that was mainly produced at the early stationary phase and reached its highest activity (3,200 AU/ml) at the same stage. It was tolerant toward a wide range of pH (2-10), temperatures (-20 to 120°C), and high concentrations of NaCl. Notably, the bacteriocin-producing strain had proteolytic activity, while the bacteriocin produced by that showed resistance to proteolytic enzymes (pepsin, trypsin, and proteinase K). Also, it was revealed that the bacteriocin activity is mostly bacteriostatic so that it considerably inhibits pathogens' growth, particularly S. aureus, E. coli, and L. monocytogenes. These characteristics prove that strain and its bacteriocin can be considered as one of the most promising agents to use in the food industry.

Antimicrobial Activity of Methanolic Extracts of Myrtus Communis L. and Eucalyptus Galbie and their Combination with Calcium Hydroxide Powder against Enterococcus Faecalis.

STATEMENT OF THE PROBLEM: The goal of endodontic therapy is the reduction or elimination of microorganisms from the root canal system. The use of intracanal medicament between treatment appointments is recommended in order to eliminate any remainang microorganisms in the pulp space. PURPOSE: The aim of the present study was to investigate the antimicrobial activity of methanolic extracts of Myrtus communis L. and Eucalyptus galbie, their combination with calcium hydroxide powder; combination of calcium hydroxide powder with water, and ready-to-use calcium hydroxide paste with iodoform; against Enterococcus faecalis. MATERIALS AND METHOD: In this experimental study, after gathering the plants, their methanolic extracts were obtained by masceration method. The diameters of inhibition zone of all mentioned materials were determined by agar diffusion test. Minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC); and anti biofilm effect of the materials that showed antibacterial effect in agar diffusion test, were then evaluated by tube dilution test, and microtiter plate assay followed by colorimetric crystal violet methods, respectively. RESULTS: After 48 hours, both herbal extracts showed antimicrobial effect. However, combination of calcium hydroxide with extracts produced no zone of inhibition. The mean inhibition zone of Eucalyptus extract was more than that of Myrtus. However, the results of ANOVA test, showed that there was no significant difference between the antibacterial effect of Eucalyptus galbie, Myrtus communis L. and positive control (Cefoxitin) (p Value= 0.987). The MIC for both extracts were 12.5 mg/ml. MBC evaluation of the two methanolic extracts showed no bactericidal effect on Enterococcus faecalis. Based on ELISA analysis, biofilm formation in response to different sub-MIC concentrations of both extracts was scored as weak to moderate. CONCLUSION: The methanolic extracts of Eucalyptus galbie and Myrtus communis L. in combination with calcium hydroxide powder were not able to eliminate Enterococcus faecalis within 48 hours.

Selective cytotoxicity of green synthesized silver nanoparticles against the MCF-7 tumor cell line and their enhanced antioxidant and antimicrobial properties.

INTRODUCTION: Silver nanoparticles (AgNPs) are of great interest due to their unique and controllable characteristics. Different synthesis methods have been proposed to produce these nanoparticles, which often require elevated temperatures/pressures or toxic solvents. Thus, green synthesis could be a replacement option as a simple, economically viable and environmentally friendly alternative approach for the synthesis of silver nanoparticles. METHODS: Here, the potential of the walnut green husk was investigated in the production of silver nanoparticles. An aqueous solution extracted from walnut green husk was used as a reducing agent as well as a stabilizing agent. Then, the synthesized nanoparticles were characterized with respect of their anticancer, antioxidant, and antimicrobial properties. RESULTS: Results showed that the synthesized nanoparticles possessed an average size of 31.4 nm with a Zeta potential of -33.8 mV, indicating high stability. A significant improvement in the cytotoxicity and antioxidant characteristics of the green synthesized Ag nanoparticles against a cancerous cell line was observed in comparison with the walnut green husk extract and a commercial silver nanoparticle (CSN). This could be due to a synergistic effect of the synthesized silver nanoparticles and their biological coating. AgNPs and the extract exhibited 70% and 40% cytotoxicity against MCF-7 cancerous cells, respectively, while CSN caused 56% cell death (at the concentration of 60 µg/mL). It was observed that AgNPs were much less cytotoxic when tested against a noncancerous cell line (L-929) in comparison with the control material (CSN). The free radical scavenging analysis demonstrated profound anti-oxidant activity for the synthesized nanoparticles in comparison with the extract and CSN. It was also detected that the synthesized AgNPs possess antibacterial activity against nosocomial and standard strains of both Gram-positive and Gram-negative bacteria (minimum inhibitory concentration =5-30 µg/mL). CONCLUSION: These findings imply that the synthesized nanoparticles using green nanotechnology could be an ideal strategy to combat cancer and infectious diseases.

A novel cationic cobalt(III) Schiff base complex: Preparation, crystal structure, Hirshfeld surface analysis, antimicrobial activities and molecular docking.

A novel Co(III) complex, [Co(L)(Imi)3]Cl incorporating 2-((3-methoxy-2-oxidobenzylidene)amino)-4-methylphenolate (L2-), as a dibasic deprotonated Schiff base ligand and imidazole (Imi) was synthesized and fully characterized using physicochemical and spectroscopic techniques including elemental analysis, conductance measurement, FT-IR, UV-Vis and X-ray single crystal diffraction. As the conductivity data showed, the synthesized complex had a 1:1 ionic nature. The structure of the complex was found to be distorted octahedral in which, O/N donor atoms of the Schiff base ligand and N atoms of three imidazole groups were involved. Antimicrobial activity of the Co(III) complex as well as the its parent Schiff base ligand against two Gram-positive bacteria (S. Aureus and M. luteus), two Gram-negative bacteria (E. coli and P. aeruginosa) and a fungus (C. Albicans) was studied. Moreover, the antimicrobial activity of [Co(L)(Imi)3]Cl was investigated using molecular docking of the complex with GlcN-6-P synthase.

Antibiotic susceptibility patterns of isolates from children with urinary tract infection in Isfahan, Iran: Impact on empirical treatment.

OBJECTIVES: The aim of this study was to identify the antibiotic susceptibility of bacteria causing urinary tract infections (UTIs) in children in Isfahan, Iran. METHODS: Retrospective and prospective analyses were conducted on isolates from children with UTIs in a referral teaching hospital of Isfahan during 2013-2015. Findings were compared between first episode versus recurrent, nosocomial versus community-acquired, previous antibiotic use versus no previous antibiotic use, and febrile versus afebrile cases. RESULTS: Among 364 patients, 68.1% had no previous UTI, 19.7% has received antibiotics prior to infection and 96.2% were infected outside the hospital. Escherichia coli was the leading cause of UTI (68.1%), followed by Enterobacter (9.3%), Klebsiella (8.8%) and other bacteria (13.7%). Most isolates were relatively highly susceptible to imipenem (79.2%), ciprofloxacin (78.0%) and nitrofurantoin (70.8%), whereas sensitivity to cefotaxime (53%), cefalexin (39.8%) and trimethoprim/sulfamethoxazole (SXT) (26.1%) was low. Resistance to imipenem, cefotaxime and cefalexin was more prevalent in recurrent cases as well as in patients consuming antibiotics prior to UTI. Resistance of nosocomial cases to nitrofurantoin and cefotaxime was higher. In addition, afebrile patients had isolates that were more resistant to imipenem, ciprofloxacin and cefotaxime. Significant cross-resistance was found between most of the studied antibiotics. CONCLUSIONS: Urinary pathogens showed low susceptibility to cefalexin, SXT and cefotaxime and high susceptibility to imipenem, ciprofloxacin and nitrofurantoin. Therefore, use of imipenem and ciprofloxacin should be considered in hospitalised children with severe or complicated disease. In addition, nitrofurantoin is a good option in afebrile cases and for UTI chemoprophylaxis in this area.

Highly Stereoselective Synthesis of Saccharin-Substituted ß-Lactams via in Situ Generation of a Heterosubstituted Ketene and a Zwitterionic Intermediate as Potential Antibacterial Agents.

Highly stereoselective synthesis of saccharin derivatives containing functionalized 2-azetidinone moiety was achieved starting from saccharin as an available precursor. The approach to these valuable heterocyclic scaffolds involves a formal [2π + 2π] cycloaddition between Schiff bases and the saccharinylketene as a novel ketene which was generated in situ and an electrocyclic reaction of a zwitterionic intermediate. The identification of the ketene was confirmed by reaction with the stable free radical TEMPO (TO•). Also, the antimicrobial activities of some new substituted saccharin against nine standard bacteria, four bacteria which were isolated from clinical samples and one yeast, were evaluated.

Synthesis, characterization, X-ray crystal structure, DFT calculation, DNA binding, and antimicrobial assays of two new mixed-ligand copper(II) complexes.

Two new Cu(II) complexes, [Cu(L)(phen)] (1), [Cu(L)(bipy)] (2), where L(2-)=(3-methoxy-2oxidobenzylidene)benzohydrazidato, phen=1,10 phenanthroline, and bipy=2,2' bipyridine, were prepared and fully characterized using elemental analyses, FT-IR, molar conductivity, and electronic spectra. The structures of both complexes were also determined by X-ray diffraction. It was found that, both complexes possessed square pyramidal coordination environment in which, Cu(II) ions were coordinated by donor atoms of HL and two nitrogens of heterocyclic bases. Computational studies were performed using DFT calculations at B3LYP/6-311+G(d,p) level of theory. DNA binding activities of these complexes were also investigated using electronic absorption, competitive fluorescence titration and cyclic voltammetry studies. The obtained results indicated that binding of the complexes to DNA was of intercalative mode. Furthermore, antimicrobial activities of these compounds were screened against microorganisms.

Detection of quinolone-resistance mutations of parC gene in clinical isolates of Acinetobacter baumannii in Iran.

BACKGROUND: The purpose of this study was to screen of parC gene mutations in clinical isolates of Acinetobacter baumannii from intensive care units (ICUs) of Alzahra Hospital, Isfahan, Iran. MATERIALS AND METHODS: Seventy isolates of A. baumannii between March 2011 and June 2012 were studied. Susceptibility test was established by E-test method. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing was performed for detection of parC gene mutation. RESULTS: 77.1% of specimens were highly resistant. Mutation at position 80 in parC was observed in 93% of isolates. CONCLUSION: High proportion of A. baumannii isolates had a mutation in parC that can play an important role in increased incidence of these isolates.

Detection of colistin sensitivity in clinical isolates of Acinetobacter baumannii in Iran.

BACKGROUND: Nosocomial infection caused by Acinetobacter baumannii has emerged as a serious problem world-wide. Finding the suitable drug is an important priority. The aim of this study was to determine colistin (polymyxin E) resistance in clinical isolates of A. baumannii from intensive care units (ICUs) of Al Zahra Hospital. MATERIALS AND METHODS: Sixty isolates of A. baumannii from patients hospitalized in ICU (Al Zahra Hospital, Isfahan University of Medical Sciences [IUMS]) were studied. All isolates of A. baumannii were tested for colistin susceptibility by Eopsilometer test (E-test). RESULTS: Of the 60 isolates 57, (95%) were multidrug resistant (MDR) and 76.6% (46/60) were highly resistant. The rate of colistin resistant with the E-test method was 11.6% (7/60). CONCLUSION: As the frequency of resistance to colistin is low, it can be used as an easily available drug for treatment of MDR A. baumannii strains, which are susceptible to colistin.

Combined experimental and theoretical studies on the X-ray crystal structure, FT-IR, 1H NMR, 13C NMR, UV-Vis spectra, NLO behavior and antimicrobial activity of 2-hydroxyacetophenone benzoylhydrazone.

A Schiff base ligand, 2-hydroxyacetophenone benzoylhydrazone (HL) was synthesized and fully characterized with FT-IR, elemental analyses, UV-Vis, (1)H NMR and (13)C NMR spectra. DFT calculations using B3LYP/6-31+G(d,p) and PW91/DZP are performed to optimize the molecular geometry. Optimized structures are used to calculate FT-IR, UV-Vis, (1)H NMR and (13)C NMR spectra of the compound. Also the energies of the frontier molecular orbitals (FMOs) have been determined. The results obtained from the optimization and spectral analyses are in good agreement with the experimental data. To investigate non-linear optical properties, the electric dipole moment (µ), polarizability (α) and molecular first hyperpolarizability (ß) were computed. The linear polarizabilities and first hyperpolarizabilities of the studied molecule indicate that the compound can be a good candidate of nonlinear optical materials. In addition, the minimal inhibitory concentration (MIC) of this compound against Staphylococcus aureus, and Candida albicans was determined.