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1.  Safety concerns to application of graphene compounds in pharmacy and medicine 
Graphene, the new allotrope of carbon is a single layer of monocrystalline graphite with sp2 hybridized carbon atoms. This compound has received worldwide attention due to its extraordinary physical and chemical properties. Duo to the widespread application of geraphenes, concerns are raising about its environmental safety or the safety protocols for handling and waste of graphene-based materials. The generation of reactive free radicals, adsorption of important biomolecules, and physical toxicity of graphene also matter. Hereby we criticize the concerns on the toxicity of graphenes to provide some perspective on the potential hazards of future development in graphene-based biomaterials.
PMCID: PMC3922742  PMID: 24450435
Graphene; Graphene oxide; Membrane; Reactive oxygen species; Safety; Toxicity
2.  Decolorization of two synthetic dyes using the purified laccase of Paraconiothyrium variabile immobilized on porous silica beads 
Decolorization of hazardous synthetic dyes using laccases in both free and immobilized form has gained attention during the last decades. The present study was designed to prepare immobilized laccase (purified from Paraconiothyrium variabile) on porous silica beads followed by evaluation of both free and immobilized laccases for decolorization of two synthetic dyes of Acid Blue 25 and Acid Orange 7. Effects of laccase concentration, pH and temperature alteration, and presence of 1-hydroxybenzotriazole (HBT) as laccase mediator on decolorization pattern were also studied. In addition, the kinetic parameters (K m and V max ) of the free and immobilized laccases for each synthetic dye were calculated.
Immobilized laccase represented higher temperature and pH stability compare to free one. 39% and 35% of Acid Blue 25 and Acid Orange 7 was decolorized, respectively after 65 min incubation in presence of the free laccase. In the case of immobilized laccase decolorization percent was found to be 76% and 64% for Acid Blue 25 and Acid Orange 7, respectively at the same time. Increasing of laccase activity enhanced decolorization percent using free and immobilized laccases. Relative decolorization of both applied dyes was increased after treatment by laccase-HBT system. After nine cycles of decolorization by immobilized laccase, 26% and 31% of relative activity were lost in the case of Acid Blue 25 and Acid Orange 7, respectively.
To sum up, the present investigation introduced the immobilized laccase of P. variabile on porous beads as an efficient biocatalyst for decolorization of synthetic dyes.
PMCID: PMC3897971  PMID: 24393474
Laccase; Immobilization; Decolorization; Synthetic dye; 1-hydroxybenzotriazole
3.  Sulfated Polysaccharides Purified from Two Species of Padina Improve Collagen and Epidermis Formation in the Rat 
Sulfated polysaccharides have shown promising effects on wound healing processes along with many other biological activities. The sulfated polysaccharides extracted from two algae species habitats in Persian Gulf were studied in vivo for their effects on collagen formation and epidermal regeneration. The polysaccharides were purified from aqueous extracts of P. tetrastromatica and P. boergesenii using CaCl2 and ethanol precipitation. The sulfate content of each polysaccharide was determined. Two identical wounds (either burn or excision) were made on the back of 4 groups of male Wistar rats (10 rats per group) under anesthesia. The algal polysaccharide ointments (2%) were applied twice daily on one side and the other wound was treated with Eucerin (as control). The rats were sacrificed on day 7 or 14, and then the wound samples were examined for epidermal thickness by light microscope. Furthermore, hydroxyproline content (as a marker of collagen formation) was spectro-photometrically measured. The polysaccharides purified from P. boergesenii had higher sulfate content (32.6±1%) compared to P. tetrastromatica (19±1%). Both algal polysaccharides showed some improvements in collagen formation (hydroxyproline content) and epidermal thickness in both wound models compared to the vehicle. The sulfated polysaccharides purified from P. tetrastromatica and P. boergesenii seaweeds are able to induce collagen formation and epidermal regeneration in the two wound models. The superior healing properties of P. boergesenii polysaccharides might be correlated to its higher sulfate content. Both algal polysaccharides are good candidates for wound healing clinical trials.
PMCID: PMC3927382  PMID: 24551807
Brown algae; fucoidan; wound healing
4.  Coumarin compounds of Biebersteinia multifida roots show potential anxiolytic effects in mice 
Traditional preparations of the root of Biebersteinia multifida DC (Geraniaceae), a native medicinal plant of Irano-Turanian floristic region, have been used for the treatment of phobias as anxiolytic herbal preparation.
We utilized the phobic behavior of mice in an elevated plus-maze as a model to evaluate the anxiolytic effect of the plant extract and bio-guided fractionation was applied to isolate the active compounds. Total root extract, alkaline and ether fraction were administered to mice at different doses 30 and 90 min prior to the maze test. Saline and diazepam were administered as negative and positive controls, respectively. The time spent in open and closed arms, an index of anxiety behavior and entry time, was measured as an index of animal activity.
The total root extract exhibited anxiolytic effect which was comparable to diazepam but with longer duration. This sustained effect of the crude extract was sustained for 90 min and was even more after injection of 45 mg/kg while the effect of diazepam had been reduced by 90 min. The anxiolytic effect factor was only present in the alkaline fraction and displayed its effect at lower doses than diazepam while pure vasicinone as the previously known alkaloid did not shown anxiolytic effect. The effect of the alkaline fraction was in a dose dependent manner starting at 0.2 mg/kg with a maximum at 1.0 mg/kg. Bio-guided fractionation using a variety of chromatographic methods led to isolation and purification of three coumarin derivatives from the bioactive fraction, including umbelliferone, scopoletin, and ferulic acid.
For the first time, bio-guided fractionation of the root extract of B. multifida indicates significant sustained anxiolytic effects which led to isolation of three coumarin derivatives with well-known potent MAO inhibitory and anti-anxiety effects. These data contribute to evidence-based traditional use of B. multifida root for anxiety disorders.
PMCID: PMC3707806  PMID: 23803166
Biebersteinia multifida; Coumarin; Anxiolytic; Scopoletin; Umbelliferone
5.  Cytotoxic evaluation of Melia azedarach in comparison with, Azadirachta indica and its phytochemical investigation 
Melia azedarach L. is an important medicinal plant that is used for variety of ailments in Iranian traditional medicine. Azadirachta indica A. Juss is its allied species and possesses similar properties and effects. The present study was undertaken to investigate anticancer activity of these M. azedarach in comparison with A. indica on cancer cell lines and also to evaluate their safety in humans by testing them on normal cell line. The study also aimed to determine the active components that are responsible for medicinal effects of M. azedarach in traditional usages.
In this study, the cytotoxic activity of crude extracts from M. azedarach and A. indica leaves, pulps and seeds as well as three main fractions of their leaf extracts were assayed against HT-29, A-549, MCF-7 and HepG-2 and MDBK cell lines. MTT assay was used to evaluate their cytotoxic activities. Methanol leaf fraction of M. azedarach as the safest leaf fraction in terms of cytotoxicity was subjected for phytochemical study.
Results of the present study indicated that seed kernel extract of M. azedarach had the highest cytotoxic activity and selectivity to cancer cell lines (IC50 range of 8.18- 60.10 μg mL-1). In contrast to crude seed extract of A. indica, crude pulp and crude leaf extracts of this plant showed remarkably stronger anti-prolifrative activity (IC50 ranges of 83.45 - 212.16 μg mL-1 and 34.11- 95.51 μg mL-1 respectively) than those of M. azedarach (all IC50 values of both plants > 650 μg mL-1). The phytochemical analysis led to the isolation of four flavonol 3-O-glycosides including rutin, kaempferol-3-O-robinobioside, kaempferol-3-O-rutinoside and isoquercetin along with a purin nucleoside, β-adenosine.
The anti-prolifrative potentials of extracts from different parts of M. azedarach and A. indica were determined. By comparison, methanol leaf fraction of M. azedarach seems to be safer in terms of cytotoxicity. Our study shows that flavonols are abundant in the leaves of M. azedarach and these compounds seem to be responsible for many of medicinal effects exploited in the traditional uses.
PMCID: PMC3664079  PMID: 23679992
Anti-prolifrative activity; Azadirachta indica; Flavonoid; Melia azedarach; MTT; Neem; Traditional medicine
6.  Biodegradation of 2,4-dinitrophenol with laccase immobilized on nano-porous silica beads 
Many organic hazardous pollutants, including 2,4-dinitrophenol (2,4-DNP), which are water soluble, toxic, and not easily biodegradable make concerns for environmental pollution worldwide. In the present study, degradation of nitrophenols-contained effluents by using laccase immobilized on the nano-porous silica beads was evaluated. 2,4-DNP was selected as the main constituent of industrial effluents containing nitrophenols. The performance of the system was characterized as a function of pH, contact time, temperature, pollutant, and mediator concentrations. The laccase-silica beads were employed in a mixed-batch reactor to determine the degradation efficiency after 12 h of enzyme treatment. The obtained data showed that the immobilized laccase degraded more than 90% of 2,4-DNP within 12 h treatment. The immobilization process improved the activity and sustainability of laccase for degradation of the pollutant. Temperatures more than 50°C reduced the enzyme activity to about 60%. However, pH and the mediator concentration could not affect the enzyme activity. The degradation kinetic was in accordance with a Michaelis–Menten equation with Vmax and Km obtained as 0.25–0.38 μmoles/min and 0.13–0.017 mM, respectively. The stability of the immobilized enzyme was maintained for more than 85% of its initial activity after 30 days. Based on the results, it can be concluded that high resistibility and reusability of immobilized laccase on CPC-silica beads make it considerable choice for wastewater treatment.
PMCID: PMC3691675  PMID: 23547870
Degradation; Laccase; Immobilization; Nano-porous silica beads; 2,4-dinitrophenol
7.  Halotolerant Ability and α-Amylase Activity of Some Saltwater Fungal Isolates  
Four halotolerant fungal isolates originating from the saltwater Lake Urmia in Iran were selected during a screening program for salt resistance and α-amylase activity. The isolates were identified based on sequencing the ITS region and a part of the β-tubulin gene, as Penicillium chrysogenum (isolate U1; CBS 132820), Fusarium incarnatum (isolate U2; CBS 132821), and Penicillium polonicum (isolate U3; CBS 132822, and isolate U4; CBS 132823). The growth of these isolates was determined by measuring the colony diameter and mycelia dry weight in Sabouraud dextrose agar and yeast nitrogen base medium supplemented with NaCl, KCl, and LiCl. Isolate U4 showed a growth up in 15% NaCl and U1 was the only isolate that could grow in 20% KCl. None of the strains grew in a media containing LiCl. The salt supplemented medium did not increase the size of colony diameter in all isolates (p > 0.05). The ability of the selected isolates for amylase production was quantitatively tested and showed that P. polonicum isolate U4 was the most potent producer of amylase with a yield of 260.9 U/L after 60 h, whereas P. polonicum isolate U3 was the lowest one with a production level of 97.9 U/L after 48 h. P. polonicum isolate U4 could be a suitable candidate for production of amylase on an industrial scale after optimization.
PMCID: PMC3813371  PMID: 24250679
Isolation; Amylase; Halotolerant; Fungi; Hypersaline environments
8.  Synthetic dye decolorization by three sources of fungal laccase 
Decolorization of six synthetic dyes using three sources of fungal laccase with the origin of Aspergillus oryzae, Trametes versicolor, and Paraconiothyrium variabile was investigated. Among them, the enzyme from P. variabile was the most efficient which decolorized bromophenol blue (100%), commassie brilliant blue (91%), panseu-S (56%), Rimazol brilliant blue R (RBBR; 47%), Congo red (18.5%), and methylene blue (21.3%) after 3 h incubation in presence of hydroxybenzotriazole (HBT; 5 mM) as the laccase mediator. It was also observed that decolorization efficiency of all dyes was enhanced by increasing of HBT concentration from 0.1 mM to 5 mM. Laccase from A. oryzae was able to remove 53% of methylene blue and 26% of RBBR after 30 min incubation in absence of HBT, but the enzyme could not efficiently decolorize other dyes even in presence of 5 mM of HBT. In the case of laccase from T. versicolor, only RBBR was decolorized (93%) in absence of HBT after 3 h incubation.
PMCID: PMC3564790  PMID: 23369690
Decolorization; Removal; Hydroxybenzotriazole; Laccase; Synthetic dyes; Oxidase
9.  Synthesis and antibacterial activity of novel levofloxacin derivatives containing a substituted thienylethyl moiety 
Background and the purpose of the study
Piperazinyl quinolones such as ciprofloxacin, ofloxacin and levofloxacin are an important group of quinolone antimicrobials which are widely used in the treatment of various infectious diseases. In the present study, we synthesized a new series of levofloxacin derivatives and evaluated their antibacterial activities.
The N-substituted analogs of levofloxacin 6a–j were prepared by nucleophilic reaction of N-desmethyl levofloxacin 11 with thienylethyl bromide derivatives 8 or 9. All target compounds were tested using conventional agar dilution method in comparison to levofloxacin and N-desmethyl levofloxacin and their MIC values were determined against a panel of Gram-positive and Gram-negative bacteria.
All compounds showed significant antibacterial activities against Gram-positive bacteria (MIC = 0.04-6.25 μg/mL); however, the activity against Gram-negative bacteria was lower (MIC = 1.56–100 μg/mL). As is evident from the data, oxime derivatives 6e, 6h and 6i are superior in inhibiting the growth of Gram-positive bacteria (MIC = 0.04–0.19 μg/mL), and their activities were found to be 5–25 times better than N-desmethyl levofloxacin 11 and equal or better than levofloxacin 4.
We have designed and synthesized novel quinolone derivatives bearing functionalized thienylethyl moiety on the piperazine ring of levofloxacin. The results of antibacterial screening against Gram-positive and Gram-negative bacteria revealed that the introduction of functionalized thienylethyl moiety on the piperazine ring of levofloxacin can improve the activity against Gram-positive bacteria. Gram-positive bacteria are responsible for a wide range of infectious diseases, and rising resistance in this group is causing increasing concern. Thus, this study introduces structural features of levofloxacin scaffold for development of new candidates in the field of anti-Gram positive chemotherapy
PMCID: PMC3807753  PMID: 23351676
Antibacterial activity; Quinolones; Levofloxacin; Thiophene derivatives; Oximes
10.  Immobilization of Laccase in Alginate-Gelatin Mixed Gel and Decolorization of Synthetic Dyes 
Alginate-gelatin mixed gel was applied to immobilized laccase for decolorization of some synthetic dyes including crystal violet. The immobilization procedure was accomplished by adding alginate to a gelatin solution containing the enzyme and the subsequent dropwise addition of the mixture into a stirred CaCl2 solution. The obtained data showed that both immobilized and free enzymes acted optimally at 50°C for removal of crystal violet, but the entrapped enzyme showed higher thermal stability compared to the free enzyme. The immobilized enzyme represented optimum decolorization at pH 8. Reusability of the entrapped laccase was also studied and the results showed that ca. 85% activity was retained after five successive cycles. The best removal condition was applied for decolorization of seven other synthetic dyes. Results showed that the maximum and minimum dye removal was related to amido black 10B and eosin, respectively.
PMCID: PMC3415199  PMID: 22899898
11.  Optimization of Chitinase Production by Bacillus pumilus Using Plackett-Burman Design and Response Surface Methodology 
A soil bacterium capable of degrading chitin on chitin agar plates was isolated and identified as Bacillus pumilus isolate U5 on the basis of 16S rDNA sequence analysis. In order to optimize culture conditions for chitinase production by this bacterium, a two step approach was employed. First, the effects of several medium components were studied using the Plackett-Burman design. Among various components tested, chitin and yeast extract showed positive effect on enzyme production while MgSO4 and FeSO4 had negative effect. However, the linear model proved to be insufficient for determining the optimum levels for these components due to a highly significant curvature effect. In the second step, Box-Behnken response surface methodology was used to determine the optimum values. It was noticed that a quadratic polynomial equation fitted he experimental data appropriately. The optimum concentrations for chitin, yeast extract, MgSO4 and FeSO4 were found to be 4.76, 0.439, 0.0055 and 0.019 g/L, respectively, with a predicted value of chitinase production of 97.67 U/100 mL. Using this statistically optimized medium, the practical chitinase production reached 96.1 U/100 mL.
PMCID: PMC3813072  PMID: 24250411
Optimization; Bacillus pumilus; Chitinase; Box-Behnken; Plackett-Burman
12.  Runx3 Expression Inhibits Proliferation and Distinctly Alters mRNA Expression of Bax in AGS and A549 Cancer Cells 
Runx3, a member of Runt-related transcription factor (Runx) proteins with tumor suppressor effect, is a tissue–restricted and cancer related transcription factor that regulate cell proliferation and growth, as well as differentiation. In the present study, exogenous Run3 was transiently expressed in AGS (human gastric adenocarcinoma), with undetectable Runx3 protein and in A549 (human lung carcinoma) with low levels of endogenous Runx3 protein.
The GFP tagged Runx3 was transfected into AGS and A549 cells using fugene6 and PolyFect and Runx3 expression was confirmed by fluorescent microscopy and RT-PCR. The effect of Runx3 transfection on cell proliferation was determined by MTT assay and the results were confirmed by the trypan blue dye exclusion method. The effect of Runx3 expression on mRNA expression of BCL2-associated X protein (Bax) was evaluated using RT-PCR.
In AGS and A549 cells, Runx3 expression inhibited cell proliferation (p < 0.01). The growth inhibition was less in A549 cells. We show that Runx3 expression increases Bax mRNA expression in AGS cells when compared with control (p < 0.05), but no significant differences in mRNA expression was observed in both examined cells.
Runx3 expression has antiproliferative effect in AGS cell perhaps via increase in expression of Bax. The effect of Runx3 on A549 cells’ viability which has endogenous level of Runx3 is not related to Bax. These findings implicate a complex regulation by Runx3 in inhibition of cell proliferation utilizing Bax.
PMCID: PMC3828922  PMID: 24250365
Runt; Cancer cells; Proliferation; Cell death; MRNA
13.  Synthesis and In-vitro Antibacterial Activities of Acetylanthracene and Acetylphenanthrene Derivatives of Some Fluoroquinolones 
Novel analogues of N-piperazinyl fluoroquinolones were prepared and evaluated against a panel of Gram-positive and Gram-negative bacteria, to study the effect of introducing bulky anthracene and phenanthrene moieties on the antibacterial effects of norfloxacin, ciprofloxacin and gatifloxacin. Although most of the novel synthesized compounds had lower antibacterial effects, some derivatives showed better activity in comparison with mother drugs based on molar concentration; for example, the 3-acetyl phenanthrene analogue of norfloxacin was more effective than E. coli and K. pneumonia.
PMCID: PMC3828925  PMID: 24250347
N-substituted piperazinyl quinolones; Anthracene derivatives; Phenanthrene derivatives; In-vitro antibacterial activity

Results 1-13 (13)