Elucidation of flavonoids from potent Iranian Scutellaria species against Influenza A (H1N1) virus.

Objectives: Influenza A virus (IAV) is a contagious illness. Different species of Scutellaria genus are used as a traditional remedy to reduce influenza symptoms. This study aimed to investigate the anti-influenza capacity of several species of Iranian Scutellaria and identify active compounds of the most potent species for the first time. Materials and Methods: Some Iranian species of Scutellaria were collected from different regions of Iran, including S. pinnatifida with mucida, viridis, and alpina subspecies; S. tournefortii; S. tomentosa; S. persica. They were fractionated to chloroform and methanol. The total phenols and flavonoids of samples were examined by the folin-ciocalteau and aluminum-flavonoid complex methods, respectively. The 50% cytotoxic concentrations (CC50) on MDCK cells and non-cytotoxic concentrations (NCTC) were determined by MTT assay. The percentage of cell protection against IAV and their effect on virus titer were investigated in pre-, post-, and co-penetration treatment groups. Phytochemicals of the most effective species were isolated by various chromatographic methods and identified by different spectroscopic methods. Results: Methanol fraction of S. pinnatifida subsp. viridis demonstrated the highest amounts of flavonoid content and best activity against influenza A virus in all combination treatments, which reduced the virus titer by 5 logs with no cytotoxicity. Kaempferol-3-O-glucoside, quercetin-3-O-glucoside, apigenin-4'-methoxy-7-O-glucoside, luteolin, and luteolin-7-O-glucoside were purified and identified from this species. Conclusion: Scutellaria pinnatifida subsp. viridis can be introduced as a source of flavonoids with acceptable anti-influenza activity. S. tomentosa also showed potent antiviral effects and is a candidate for elucidation in further studies.

99mTc- Anionic dendrimer targeted vascular endothelial growth factor as a novel nano-radiotracer for in-vivo breast cancer imaging.

Since breast cancer is the commonly cause of death among women around the world, diagnosis at the early stages is significantly important to prevent the metastasis of the cancer. Among the various growth factors that are involved in angiogenesis, vascular endothelial growth factor (VEGF) is believed to be the most important factor. Overexpressed VEGF receptor on tumors surface, is particularly interesting for cancer cells targeting purposes. In this study, citric acid dendrimer conjugated with VEGF antagonist peptide was synthesized. The obtained product was confirmed by FT-IR, TEM, DLS, and EDS. In vitro cytotoxicity assay showed no toxicity on normal cells and indicated the notably dose-dependence toxicity on cancer cells. Box-Behnken software as a computational method was used to determine the optimum amount of radiolabeling parameters. Optimized parameters for reducing agent, dendrimer-anti-VEGF, and time were 1.4 mg, 17.5 mg, and about 30 min respectively. Radiochemical purity of radio-labeled conjugated dendrimer was determined about 90 percent. SPECT imaging was done to observe the in vivo accumulation of dendrimer-anti-VEGF in the tumor site. Images showed high accumulation of radio-tracer in the tumor region. All in all, obtained results confirmed our hypothesis that the dendrimer-anti-VEGF can be a good radio-tracer for diagnosis of cancer.

N-acetylcysteine-PLGA nano-conjugate: effects on cellular toxicity and uptake of gadopentate dimeglumine.

Gadolinium as a contrast agent in MRI technique combined with DTPA causes contrast induced nephropathy (CIN) and nephrogenic systemic fibrosis (NSF) which can reduce by usage of antioxidants such as N-acetyl cysteine by increasing the membrane's permeability leads to lower cytotoxicity. In this study, N-acetyl cysteine-PLGA Nano-conjugate was synthesized according to stoichiometric rules of molar ratios andafter assessment by FTIR, NMR spectroscopy and Atomic Force Microscopy (AFM) imaging was combined with Magnevist® (gadopentetate dimeglumine) and its effects on the renal cells were evaluated. MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide] and cellular uptake assays have indicated relatively significant toxicity of magnevist (P < 0.05) on three cell lines including HEK293, MCF7 and L929 compared to other synthesized ligands that shown no toxicity. Moreover, systemic evaluation has shown no notable changes of blood urea nitrogen (BUN) and creatinine in kidney of mice. In consequence, antioxidant effect was increased as well as the renal toxicity of the contrast agent reduced at the cell level. As a result, PLGA-NAC nano-conjugate can be a promising choice for decreasing the magnevist toxicity for treatment and prevention of CIN and will be able to open a new horizon to research on reduction of toxicity of contrast agents by using nanoparticles.

Exposure Assessment for Some Pesticides through Rice Consumption in Iran Using a Multiresidue Analysis by GC-MS.

In communities which consume rice as main food, importance of risk assessment for contaminants is always taken into consideration by health authorities. The present study is an attempt for monitoring of 56 pesticides from different chemical groups in rice samples collected from local markets in Tehran and estimation of daily intake of interested pesticides through this monitoring. A valid method based on spiked calibration curves and QuEChERS sample preparation was developed for determination of pesticides residue in rice by GC/MS. The analytical results of the proposed method were in good agreement with the proficiency test (FAPAS 0969). One-hundred-thirty-five rice samples were analyzed and 11 pesticide residues were found in 10.4% of the samples. Of which 5.2% were contaminated with unregulated pesticides. None of the samples, which were contaminated with regulated pesticides, had contamination higher than maximum residue limit. The mean estimated dose (ED) was calculated with respect of mean of contamination and mean daily consumption of rice. ED of the found pesticides is much lower than the related ADIs.

Novel chlorambucil-conjugated anionic linear-globular PEG-based second-generation dendrimer: in vitro/in vivo improved anticancer activity.

Evaluating the efficacy of anticancer drugs is an evolving and research-oriented issue. The objective of this study was to reduce the insolubility of chlorambucil (CBL) in water and improve the anticancer activity of CBL in vitro and in vivo through the conjugation of CBL with anionic linear-globular dendrimer (second generation, G2). In the current study, the anticancer activity among three groups that include CBL, CBL-G2 dendrimer, and control was measured in vitro and in vivo. In vitro studies showed that G2 anionic linear-globular polyethylene-glycol-based dendrimer, which conjugated to the CBL exterior through an ester linkage, was able to significantly improve the treatment efficacy over clinical CBL alone with respect to proliferation assay, 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide; half maximal inhibitory concentration (IC50) was calculated to be 141 µg/mL for CBL alone and 27.7 µg/mL for CBL-G2 dendrimer; P<0.05. In addition, CBL-G2 dendrimer conjugate forestalled the growth of MCF-7 cancerous cells in addition to enhancing the number of apoptotic and necrotic cells as demonstrated by an annexin V-fluorescein isothiocyanate assay. CBL-G2 dendrimer conjugate was able to checkmate antiapoptotic Bcl-2 expression and Bcl-2/Bax ratio in a large scale compared with the control group and CBL alone (P<0.005). In vivo studies showed that tumor treatment by CBL-G2 dendrimer conjugate outstrips the efficacy of treatment compared with CBL alone. The evaluation was based on reduction in tumor volume and tumor growth inhibition of murine 4T1 mammary tumor cells. Tumor volume of 140%±8% was measured in the treatment with CBL-G2 dendrimer, whereas 152%±13.5% was calculated in the treatment with free CBL (P<0.05). However, there were no significant differences in histological assay among the three groups. In conclusion, tumor growth suppression potential of CBL-G2 dendrimer, which was assessed in both in vitro and in vivo experiments, has provided empirical evidence to buttress the fact that this compound could be considered for functional cancer treatment with low side effects.

Synthesis and Evaluation of Chroman-4-One Linked to N-Benzyl Pyridinium Derivatives as New Acetylcholinesterase Inhibitors.

A novel series of chroman-4-one derivatives containing the N-benzyl pyridinium moiety were designed, synthesized, and evaluated for their acetylcholinesterase (AChE) inhibitory activities. Among the various synthesized compounds, (E)-1-(2,3-dibromobenzyl)-4-((7-ethoxy-4-oxochroman-3-ylidene)methyl)pyridinium bromide (8l) depicted the most potent anti-AChE activity (IC50 = 0.048 µM). In addition, the molecular modeling study allowed us to detect possible binding modes that are in full compliance with the observed results through in vitro experiments.

New tetracyclic tacrine analogs containing pyrano[2,3-c]pyrazole: efficient synthesis, biological assessment and docking simulation study.

A new series of tacrine-based acetylcholinesterase (AChE) inhibitors 7a-l were designed by replacing the benzene ring of tacrine with aryl-dihydropyrano[2,3-c]pyrazole. The poly-functionalized hybrid molecules 7a-l were efficiently synthesized through multi-component reaction and subsequent Friedländer reaction between the obtained pyrano[2,3-c]pyrazoles and cyclohexanone. Most of target compounds showed potent and selective anti-AChE activity at sub-micromolar range. The most potent compound 7h bearing a 3,4-dimethoxyphenyl group was more active than reference drug tacrine. The representative compound 7h could significantly protect neurons against oxidative stress as potent as quercetin at low concentrations. The docking study of compound 7h with AChE enzyme revealed that the (R)-enantiomer binds preferably to CAS while the (S)-enantiomer prone to be a PAS binder.

Simultaneous Determination of 17 Pesticide Residues in Rice by GC/MS using a Direct Sample Introduction Procedure and Spiked Calibration Curves.

A reliable, rapid and accurate method based on spiked calibration curves and direct sample introduction was developed for determination of 17 pesticide residues in rice by gas chromatography-mass spectrometry single quadrupole selected ion monitoring GC/MS-SQ-SIM. Sample preparation is based on extraction with acetonitrile without clean up. The use of spiked calibration curves for constructing the calibration curve substantially reduced adverse matrix-related effects. The average recovery of pesticides at 6 concentration levels was in range of 97.5-102.1%. The method was proved to be repeatable with RSDr in range of 0.7%-19.8%for all of the concentration levels. The limits of detection and limit of quantifications for all the pesticides were < 10 ng/g and < 25 ng/g, respectively. The developed method was applied for simultaneous determination of the selected pesticides in 23 rice samples collected from Tehran retail market in March 2009. Although many studies have been conducted regarding the determination of pesticides by using GC-MS, this is the first attempt in Iran using GC-MS-SIM technique that successfully can determine 17 pesticides with difference in physicochemical properties in rice.

5,6-Dimethoxybenzofuran-3-one derivatives: a novel series of dual Acetylcholinesterase/Butyrylcholinesterase inhibitors bearing benzyl pyridinium moiety.

BACKGROUND: Several studies have been focused on design and synthesis of multi-target anti Alzheimer compounds. Utilizing of the dual Acetylcholinesterase/Butyrylcholinesterase inhibitors has gained more interest to treat the Alzheimer's disease. As a part of a research program to find a novel drug for treating Alzheimer disease, we have previously reported 6-alkoxybenzofuranone derivatives as potent acetylcholinesterase inhibitors. In continuation of our work, we would like to report the synthesis of 5,6-dimethoxy benzofuranone derivatives bearing a benzyl pyridinium moiety as dual Acetylcholinesterase/Butyrylcholinesterase inhibitors. METHODS: The synthesis of target compounds was carried out using a conventional method. Bayer-Villiger oxidation of 3,4-dimethoxybenzaldehyde furnished 3,4-dimethoxyphenol. The reaction of 3,4-dimethoxyphenol with chloroacetonitrile followed by treatment with HCl solution and then ring closure yielded the 5,6-dimethoxy benzofuranone. Condensation of the later compound with pyridine-4-carboxaldehyde and subsequent reaction with different benzyl halides afforded target compounds. The biological activity was measured using standard Ellman's method. Docking studies were performed to get better insight into interaction of compounds with receptor. RESULTS: The in vitro anti acetylcholinesterase/butyrylcholinesterase activity of compounds revealed that, all of the target compounds have good inhibitory activity against both Acetylcholinesterase/Butyrylcholinesterase enzymes in which compound 5b (IC50 = 52 ± 6.38nM) was the most active compound against acetylcholinesterase. The same binding mode and interactions were observed for the reference drug donepezil and compound 5b in docking study. CONCLUSIONS: In this study, we presented a new series of benzofuranone-based derivatives having pyridinium moiety as potent dual acting Acetylcholinesterase/Butyrylcholinesterase inhibitors.

Carcinoembryonic antigen admittance biosensor based on Au and ZnO nanoparticles using FFT admittance voltammetry.

In this work, a highly sensitive carcinoembryonic antigen fast Fourier transform admittance biosensor is introduced. The proposed biosensor is based on bilayer films of ZnO/Au nanoparticles as an immobilization matrix. These layers are prepared by self-assembly and deposition method on a gold electrode surface, respectively. Carcinoembryonic antibody (anti-CEA) was immobilized on gold nanoparticles and positively charged horseradish peroxidase (HRP) was used to block sites against nonspecific binding. The admittance biosensor was developed based on fast Fourier transform continuous square wave voltammetry, which produces a sensitive, fast (less than 20 s) and reliable response for determination of carcinoembryonic antigen. The technique was applied as a detector in a flow injection system. The admittances reduction current of the biosensor decreases linearly in two concentrations ranges of CEA from 0.1 to 70 ng/mL and from 70 to 200 ng/mL with a detection limit of 0.01 ng/mL in presence of 0.5 mM H(2)O(2) as an eluent solution.

Effect of cooking process on the residues of three carbamate pesticides in rice.

A gas chromatography mass spectrometry with spike calibration curve method was used to quantify three carbamate pesticides residue in cooked white rice and to estimate the reduction percentage of the cooking process duration. The selected pesticides are three carbamate pesticides including carbaryl and pirimicarb that their MRL is issued by "The Institute of Standards of Iran" and propoxur which is used as a widely consumed pesticide in rice. THE ANALYTICAL METHOD ENTAILED THE FOLLOWING STEPS: 1- Blending 15 g cooked sample with 120 mL acetonitrile for 1 min in solvent proof blender, 2- Adding 6 g NaCl and blending for 1 min, 3- Filtering upper layer through 25 g anhydrous Na2SO4, 4- Cleaning up with PSA and MgSO4, 5- Centrifuging for 7 min, 6- Evaporating about 0.3 mL and reconstituting in toluene till 1 mL, 7- Injecting 2 µL extract into GC/MS and analyzing by single quadruple selected ion monitoring GC/MS-SQ-SIM. The concentration of pesticides and the percentage of pesticides amounts after the cooking were determined by gas chromatography mass-spectrometry (GC/MS) using with interpolation of the relative peak areas for each pesticide to internal standard peak area in the sample on the spiked calibration curve. Calibration curve was linear over the range of 25 to 1000 ng/g, and LOQ was 25 ng/g for all three pesticides. The percent of loss for the three pesticides were 78%, 55% and 35% for carbaryl, propoxur and pirimicarb respectively. Different parameters such as vapor pressure, boiling point, and suspect ability of the compound to hydrolysis, could be responsible for the losing of pesticides during the cooking process.

Design, synthesis and anticholinesterase activity of a novel series of 1-benzyl-4-((6-alkoxy-3-oxobenzofuran-2(3H)-ylidene) methyl) pyridinium derivatives.

A novel series of benzofuranone-ylidene-methyl benzylpyridinium derivatives (6a-u) were synthesized as acetylcholinesterase inhibitors. The anticholinesterase activity of synthesized compounds was measured using colorimetric Ellman's method. It was revealed that some synthesized compounds exhibited high anticholinesterase activity, among them compound 6b was the most active compound (IC(50)=10 + or - 6.87 nM).

Determination of morphine in the plasma of addicts in using Zeolite Y extraction following high-performance liquid chromatography.

BACKGROUND: The measurement of morphine in biological samples has become a routine in many clinical and forensic toxicology laboratories. A high-performance liquid chromatography (HPLC) method was developed for the determination of morphine in plasma. METHODS: Samples were extracted using Zeolite Y column followed by reversed phase HPLC with fluorescence detection. This method was based on an ex-calibration procedure and was linear between 20 and 200 ng/ml of morphine. Blood from 10 male opiate addicts were obtained from Rosbeh Hospital. All of the male smoked opiate (heroin, opium) and cigarettes. RESULTS: The mean total level 5 h after the last abuse was 152.4 ng/ml and 37.6 ng/ml at 10-15 h. The method was reliable for morphine determination in blood even after 5 half-lives after the last abuse. CONCLUSIONS: This method is simple and rapid and may be useful for routine monitoring of plasma morphine concentration.