Iranian Journal of Pharmaceutical Research
Volume:12 Issue: 4, Autumn 2013

Transgenic (Tg) mice are very important models of disease, and have been introduced to biological studies since 1982. They are used for understanding the pathobiology of different diseases, finding targets for pharmacological manipulations, and for the evaluation of efficacy and toxicity of medicines in preclinical studies. Many different types of Tg mice are available as models for human diseases. For examples, there are Tg mice for different types of cancers, Down syndrome, Alzheimer’s disease, Huntington›s disease, diabetes mellitus, heart disease, hypertension, epilepsy, obesity, osteoporosis, glaucoma, blindness, deafness, anxiety, depression, and many other diseases. Transgenic animals are used in toxicity assessments as well. There are several Tg animal models for mutagenicity assays approved by the World Health Organization like lacZTg mice for evaluation of mutagenicity of medicines. Tg-rasH2 mouse is another Tg model sensitive to both genotoxic and non-genotoxic carcinogens and is used in pharmaceutical industries as a substitute method to evaluate the carcinogenicity of novel compounds. In recent years the global attention to biotechnological products has been dramatically increased. Transgenic animals can also be used to produce recombinant proteins.There are many examples of such applications for Tg animals. Productions of tissue plasminogen activator and recombinant human antithrombin III in goat are cases of using Tg animals in the biotech industries. Production of Factors VIII and IX and protein C in cows, or developing sheep milk includes thrombin and Factor XIII, and alpha-1-antitrypsin, or expression of malaria protein in mice for vaccine development are important examples for this kind of application. The ethical concerns about the genetic modifications of animals are very important issues and have been closely debated in many scientific discussions. In the last decade the contribution of Iranians in science production and their technological achievements, especially in medical and biological sciences, have been considerably increased. Therefore, demanding a greater consideration for genetically modified animals. Several strains of Tg mice are commercially available in many countries. However, the access to these animals for pharmaceutical researchers is very limited at this time in Iran. This requires serious attention from decision makers in the medical researches in Iran in order for this problem to be solved. Commercially available Tg animals can be provided either by domestic or international private companies and will be a great service for scientists and researchers in Iran.

In this paper, carbon nanotubes (CNTs) were added into chitosan (CS) hydrogels in the form of chitosan modified CNTs (CS-CNTs) composites to prepare carbon nanotubes hydrogels (CNTs-GEL). The products, named CS-MWCNTs, were characterized by scanning electron microscope (SEM) and Fourier transform infrared (FTIR) spectroscopy. Swelling properties and effect of pH on controlled release performance of the two kinds of hydrogels, CNTs-GEL and pure chitosan hydrogels without CNTs (GEL), were investigated respectively. The results showed that CNTs-GEL possess better controlled release performance than GEL. The releasing equilibrium time of CNTs-GEL was longer than that of GEL in both pH = w7.4 and pH=1.2 conditions, although the release ratios of the model drug are similar in the same pH buffer solutions. It is found that release kinetics is better fitted Ritger-Peppas empirical model indicating a fick-diffusion process in pH = 1.2, while in pH = 7.4 it was non-fick diffusion involving surface diffusion and corrosion diffusion processes.

Clarithromycin (CLA), a broad-spectrum macrolide, is a poorly soluble drug with dissolution rate limited absorption. The aim of this investigation was to prepare CLA nanoparticles from a ternary ground mixture in the presence of sodium lauryl sulfate (SLS) and polyvinyl pyrrolidone (PVP) as co-grinding water-soluble compounds, in order to improve the drug dissolution rate. Different weight ratios of CLA: SLS: PVP were ground in a dry process by planetary ball mill using different grinding ball size. Following the dissolution rate study, physical properties of the best dissolved co-ground formulation was studied. The accelerated stability studies were also conducted on the co-ground formulation. The results revealed that the dissolution rate of ternary ground mixtures was much higher than that of the intact drug (p < 0.001). Decreasing the grinding ball size and weight with the same rotation speed resulted in particles with decreased dissolution. On the other hand, increasing the PVP concentration in the formulations reduced the drug dissolution. Dissolution efficiencies (DE10 and DE30) for the best dissolved formulation, which consisted of the equal ratio of each co-ground component, were 8.7 and 5 folds higher than the untreated CLA, respectively. This formulation formed nanocrystals with enhanced solubility after dispersing in water. X-ray diffraction, differential scanning calorimetry and infrared spectrophotometry confirmed no chemical interaction and phase transition during the process. Accelerated stability studies confirmed that the co-ground mixture almost remained unchanged in terms of dissolution rate, drug assay and particle size after exposing in stability conditions for three months.

In this study, tretinoin microemulsion has been formulated based on phase diagram studies by changing the amounts and proportions of inactive ingredients, such as surfactants, co-surfactants and oils. The effects of these variables have been determined on microemulsion formation, particle size of the dispersed phase and release profile of tretinoin from microemulsion through dialysis membrane. In released studies, static Franz diffusion cells mounted with dialysis membrane were used. Sampling was conducted every 3 h at room temperature over a period of 24 h. The amount of released drug was measured with UV-spectrophotometer and the percentage of drug released was calculated. Based on the results obtained, the oil phase concentration had a proportional effect on particle size which can consequently influence on drug release. The particle size and the amount of released drug were affected by the applied surfactants. The components of the optimized microemulsion formulation were 15% olive oil, 12% propylene glycol (as co-surfactant), 33% Tween®80 (as surfactant) and 40% distilled water, which was tested for viscosity and rheological behavior. The prepared tretinoin microemulsion showed pseudoplastic-thixotropic behavior. The profile of drug release follows zero order kinetics. The optimized tretinoin microemulsion showed enhanced in-vitro release profile compared to the commercial gels and creams.

Cefquinome Sulfate (CS) is a fourth-generation cephalosporin, which has been developed solely for veterinary use. It shows potent antibacterial activity against a broad spectrum of bacterial species. However, Cefquinome is susceptible to hydrolysis, which limiting its clinical employment efficacies to some extent. So, in this study, to increase Cefquinome Sulfate biological half-life, a novel Cefquinome Sulfate proliposome was prepared by solid dispersion and effervescent techniques and characterized for morphology, particle size, entrapment efficiency and in vitro release. A Reversed Phase-High Performance Liquid Chromatography (RP–HPLC) method was first chosen and established to determine the drug concentration in plasma after intra muscular (IM) administrating Cefquinome Sulfate solution and liposome at a single dosage of 18 mg/kg in rabbit. Then their pharmacokinetics in vivo was compared. Results showed that the received liposome was milky white suspension, spherical or ellipsoidal in shape. The mean particle size was 203±5 nm and the entrapment efficiency was 53.5±0.16%. The cefaquinom sulfate solution and liposome both followed a two compartment model, in vivo. The pharmacokinetic parameters for the solution and liposomal formulations were measured as follows: t1/2α were (1.214 ± 0.135) h and (1.395 ± 0.113) h, t1/2β were (8.752 ± 0.846) h and (16.503 ± 1.275) h, AUC(0-24) were (49.582 ± 9.173) (mg·h)/L and (138.727 ± 11.034) (mg·h)/L, CL/F were (0.357 ± 0.015) L/(h·kg) and (0.127 ± 0.012) L/(h·kg), MRT(0-24) were (2.68 ± 0.229) h and (5.945 ± 0.479) h, respectively. It could be clearly seen that t1/2β of liposome prolonged (p < 0.05), AUC and MRT both increased remarkably (p < 0.01), CL/F decreased. Results indicated that this preparation has more residence time and exhibits some sustained–release tendency.

Attempts have been made to prepare nanoparticles based on poly(lactic-co-glycolic acid) (PLGA) and doxorubicin. Biological evaluation and physio-chemical characterizations were performed to elucidate the effects of initial drug loading and polymer composition on nanoparticle properties and its antitumor activity. PLGA nanoparticles were formulated by sonication method. Lactide/glycolide ratio and doxorubicin amounts have been tailored. Fourier transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC) were employed to identify the presence of doxorubicin within nanospheres. The in vitro release studies were performed to determine the initial ant net release rates over 24 h and 20 days, respectively. Furthermore, cytotoxicity assay was measured to evaluate therapeutic potency of doxorubicin-loaded nanoparticles. Spectroscopy and thermal results showed that doxorubicin was loaded into the particles successfully. It was observed that lactide/glycolide content of PLGA nanoparticles containing doxorubicin has more prominent role in tuning particle characteristics. Doxorubicin release profiles from PLGA 75 nanospheres demonstrated that the cumulative release rate increased slightly and higher initial burst was detected in comparison to PLGA 50 nanoparticles. MTT data revealed doxorubicin induced antitumor activity was enhanced by encapsulation process, and increasing drug loading and glycolide portion. The results led to the conclusion that by controlling the drug loading and the polymer hydrophilicity, we can adjust the drug targeting and blood clearance, which may play a more prominent role for application in chemotherapy.

Losartan potassium, Valsartan, Telmisartan and Irbesartan are angiotensin-II-receptor antagonists (ARA II) group which used in treatment of hypertension alone or in combination with other drugs mainly Hydrochlorothiazide. RP- HPLC method was developed for the assay of three angiotensin-II-receptor antagonists (ARA-IIs) in presence of Hydrochlorothiazide. The method was performed by reversed phase high performance liquid chromatography using a mobile phase which is consisted of 0.025 M potassium dihydrogen phosphate (pH 6.0): acetonitrile =65:35% with detection at 220 nm on an ACE C18 column (250 mm × 4.6 mm, 5 μm) at flow rate 1.5 ml/min in an isocratic manner. The proposed method was validated according to ICH guidline in terms of linearity, accuracy, precision, robustness, limit of detection and limit of quantitation.

A novel potentiometric ion-selective PVC membrane sensor for analysis of atorvastatin (AT) in pharmaceutical preparations based on atorvastatin-(tetraphenyl borate), (AT-(TPB)2) as sensing element, tetraphenyl borate as additive and tris-2-ethyl-hexyl phosphate (TOP) as plasticizer solvent was prepared. The electrode shows a good Nernestian response over the concentration range of 0.09–5586 µg mL-1of AT with slope of 30.1±0.1 mV/decade and limit of detection0.056µg mL-1.The response time of sensor is fats (less than 10 sec)and could be used for about one month in the pH range of 4.5–8.0. The electrode exhibit good selectivity for the AT in the presence of large amount of co-drugs and inorganic cations. The method is precise and accurate with mean relative standard deviation of <2%.Atorvastatin is determined successfully in several tablets by the proposed membrane.

The forced degradation study of lidocaine HCl was carried out according to the ICH guideline Q1A (R2). The degradation conditions were assessed to be hydrolysis, oxidation, photolysis and dry heat during 24 h, 48 h and 72 h and then the samples were investigated by GC-FID method and nuclear magnetic resonance (NMR) spectroscopy. According to these results, the degradation products were not observed in all reaction conditions during the 72 h period. Only spectral changes in the 1H and 13C-NMR spectrum were observed in hydrogen peroxide and acid degradation. As a result of this degradation, n-oxide was formed. After acid-induced degradation with HCl, the secondary amine salt was formed. Furthermore, trifluoroacetic acid (TFA) was used as the acidic media, and the decomposition products were observed. A simple and reliable gas chromatography method with flame ionization detection (GC-FID) was developed and validated for the determination of lidocaine HCl in pharmaceutical formulations in the form of a cream and injections. The GC-FID method can be used for a routine analysis of lidocaine HCl in pharmaceutical formulations and the proposed method, together with NMR spectroscopy, can be applied in stability studies.

In this work, we developed a simple and selective method for separation and spectrophotometric determination of trace amounts of cetylpyridinium chloride (CPC) in pharmaceutical products using cloud point extraction (CPE) technique. The method is based on cloud point extraction of the CPC in alkali conditions using of nonionic surfactant Triton X-114. Under selected conditions, the calibration graph was linear in the range of 0.50-30 µg/mL of CPC with r=0.9993 (n=10). Average recoveries for spiked samples were determined to be between 95–104%. The relative standard deviation (RSD) for 5.0 µg/mL of CPC was 1.86 % (n=10). Also, the use of micellar extraction for extracting CPC was enhanced the molar absorptivity (ε) from 1.83103 L/mol.cm in aqueous solution to 1.539104 L/mol.cm in surfactant-rich phase. The proposed method was applied for the determination of CPC in a commercial mouth washer product

A series of 2-amino-4-(nitroalkyl)-4H-chromene-3-carbonitriles were synthesized by an efficient multicomponent reaction in aqueous media using DBU (1,8-diazabicyclo[5.4.0]undec-7-ene) as a catalyst at room temperature. Mild condition, environment friendly procedure and excellent yields are the main advantages of this procedure. The cytotoxic activity of target compounds were evaluated against three cancer cell lines MDA-MB-231, MCF-7 and T47D in comparison with etoposide as reference drug. Generally, all compounds showed good cell growth inhibitory activity with IC50 values less than 30 μg/mL. Their activities were comparable or more potent than standard drug etoposide. The 6-bromo- derivatives 7e and 7f showed promising cytotoxic activity with IC50 values in the range of 3.46–18.76 μg/mL, being more potent than etoposide against all tested cell lines.

Cancer is the second leading cause of death in the world. Despite advances in the diagnosis and treatment, overall survival of patients still remains poor. Hence, there is an urgent need for development of new anticancer agents. Considering promising biological activity of 1,3,4-thiadiazole derivatives, in the present study, synthesis and cytotoxicity assessment of new derivatives of this ring was done. All synthesized compounds were characterized by NMR, IR and MS spectroscopic methods. Obtained data from MTT assay showed that all compounds 3a-3l hadbetter anticancer activity against MDA(breast cancer) compared to PC3(prostate cancer) and U87(Glioblastoma).Compound 3g with m-OCH3 moiety on the phenyl ring was the most potent one in this series with IC50 = 9 µM against MDA breast cell line in comparison with imatinib(IC50 = 20 µM) as reference drug.

Compounds containing triazene ring structure are cytotoxic agents and clinically used as antitumor alkylating agents. In this study, a series of triazene derivatives holding alkyl and aryl moieties were synthesized and proved to be potent cytotoxic agents in-vitro particularly against eight cancer cell lines (PC3, HT29, Hela, HL60, Jurkat, K562, MCF7, HepG2) and a non-cancerous cell line (HUVEC). The cytotoxic activity was assessed using two methods, LDH assay, and trypan blue exclusion. Some of the triazene derivatives showed cytotoxic activity more than temozolomide (TMZ) as the reference drug. The synthesized triazenes showed marked cytotoxicity effects on all eight cancer cell lines. Among the compounds synthesized, 1,3-bis(2-ethoxyphenyl)triazene C had unique efficacy and selectivity so that it had IC50 between 0.560-3.33 µM on cancer cell lines and 12.61 μM on normal cell line (HUVEC). 1-(4-nitrophenyl)-3-(2-hydroxyethyl)triazene E shows weaker effect on cancer cell lines than the other compounds having IC50 between 3-15.54 μM.

Caffeic acid phenethyl ester(CAPE) suppresses the growth of transformed cells such as human breast cancer cells, hepatocarcinoma, myeloid leukemia, colorectal cancer cells, fibrosarcoma, glioma and melanoma. A group of heterocyclic esters of caffeic acid was synthesized using Mitsunobu reaction and the esters were subjected to further structural modification by electrooxidation of the catechol ring of caffeic acid esters in the presence of sodium benenesulfinate and sodium toluensulfinate as nucleophiles. Both heterocycle esters of caffeic acid and their arylsulfonyl derivatives were evaluated for their cytotoxic activity against HeLa, SK-OV-3, and HT-29 cancer cell lines. HeLa cells showed the highest sensitivity to the compounds and heterocyclic esters with no substituent on catechol ring showed better activity compared to their substituted counterparts. QSAR studies reemphasized the importance of molecular shape of the compounds for their cytotoxic activity.

The uses of non-steroidal anti-inflammatory drugs (NSAIDs) are limited by a variety of side effects. So research on preparing new analgesic agents is important. According to some reports about the analgesic activity of hydrazide and hydrazine derivatives a new series of these compounds were synthesized in order to obtain new analgesic compounds. The final compounds 10a-10e and 15a-15d were prepared by condensation of corresponding hydrazides 7 and 8with different aldehydes 9a-9e. The structures of all synthesized compounds were confirmed by means of FT-IR, 1H-NMR and Mass spectra. All compounds were evaluated for their analgesic activities by abdominal constriction test (writhing test). Most of the synthesized compounds induced significant reduction in the writhing response when compared to control and compound 15was more potent than mefenamic acid in the writhing test.

Radioiodinated meta iodobenzylguanidine (MIBG) is one of the important radiopharmaceuticals in Nuclear Medicine. [123/131I]MIBG is used for imaging of Adrenal medulla, studying heart sympathetic nerves, treatment of pheochromacytoma and neuroblastoma. For clinical application, radioiodinated MIBG is prepared through isotopic exchange method, which includes replacement of radioactive iodine in a nucleophilic substitution reaction with cold iodine (127I). The unlabelled MIBG hemisulfate is synthesized by the procedure described by Wieland et al (1980). The availability of a more practical and cost-effective procedure for MIBG preparation encouraged us to study the MIBG synthesis methods. In this study the preparation of MIBG through different methods were evaluated and a new method, which is one step, simple and cost-effective is introduced. The method has ability to be scaled up for production of unlabelled MIBG.

Radiolabeled porphyrins are potential tumor avid radiopharmaceuticals because of their impersonation in the human body, ability to complex various radionuclides, water solubility, low toxicity etc. in this work a radiogallium porphyrin complex has been developed. [67Ga] labeled 5,10,15,20-tetrakis(3,4-dimethoxyphenyl) porphyrin ([67Ga]-TDMPP) was prepared using freshly prepared [67Ga]GaCl3 and 5,10,15,20-tetrakis(3,4-dimethoxyphenyl) porphyrin (H2TDMPP) for 60 min at 100C (. Stability of the complex was checked in final formulation and human serum for 24 h, followed by biodistribution and imaging studies in wild type rats up to 24 h. The radiocomplex was obtained with radiochemical purity >99% (ITLC) and >98% (HPLC), specific activity: 12-15 GBq/mmol. The partition coefficient was determined (log P=1.63). A detailed comparative pharmacokinetic study performed for 67Ga cation and [67Ga]-TDMPP. The complex is mostly washed out from the circulation through kidneys. Myocardial uptake was significantly observed by SPECT and biodistribution studies. Knee and shoulder joints demonstrated significant activity uptake in 2h post injection. Higher water solubility of the complex due to ionic nature of the complex is an advantage for rapid wash-out of the complex from the system, the complex has significant joint uptake compared to other radiolabeled porphyrins which the mechanisms are explained.

Pyrazine derivatives are important class of compounds with diverse biological and cytotoxic activities and clinical applications. In this study, B3 p 86 / 6 – 31 + + G * was used to compute and map the molecular surface electrostatic potentials of a group of substituted amides of pyrazine-2-carboxylic acids to identify common features related to their subsequent cytotoxicities. Several statistical properties including potentials extrema (Vs, min,Vs, max), the average of positive electrostatic potential on the surface (Vs+), the average of V(r) over the surface (Vs) and the Lowest Unoccupied Molecular Orbital (LUMO) and system cytotoxicities were computed. Statistically, the most significant correlation is a five -parameter equation with correlation coefficient, R² values of 0.922 and R²adj = 0.879. The obtained models allowed us to reveal cytotoxic activity of substituted amides of Pyrazine2- carboxcylic acid.

Sesame (Sesamum indicum L.) seed and oil have long been used widely as healthy foods to supply energy and prevent aging. Some of the main active anti-oxidative constituents in sesame seeds are γ-tocopherol and phenols. The purpose of this study was to investigate the relationship between roasting temperature and time with γ-tocopherol and total phenolic compounds (TPC) of sesame seeds when roasted in a domestic electric oven. Eight cultivars of sesame seeds in this study were Darab, Dezful, Karaj, Moghan, Naz-Branching, Naz-NonBranching, Siah and Varamin. Each cultivar was divided into ten group based on the roasting time (10, 15 and 20 min) and temperatures (180, 200 and 220°C)andunroasted one. The high-performance liquid chromatography (HPLC) and spectrophotometeric methods were used for γ-tocopherol (n = 80) and TPC (n = 80) analysis, respectively. The γ-tocopherol content ranged from 329 ± 5 mg/L in Naz-Branching sesame oil to 1114±7 mg/L in Siah sesame oil and 169±6 to 577±1 mg/kg in sesame seed respectively. γ-tocopherol content of six cultivars increased significantly (p < 0.05) as the roasting temperature and time; until 200°C for 10 min, but they were decreased by roasting at 220 °C in longer time. Also TPC increased significantly as the roasting temperature. The amount of TPC varied in different sesame cultivars from 20.109 ± 3.967 µM to 129.300±3.493 in Varamin and Naz-Branching sesame seed cultivars, respectively, also TPC increased from 70.953 ± 5.863 µM in unroasted Naz-Branching sesame seed to 129.300 ± 3.493 µM after roasting in 200°C for 20 min. The present study showed that Iranian sesame seed can be considered as a good source of natural antioxidant specially after roasting. The optimum temperature and time roasting to obtain the most γ-tocopherol and total phenolic content was 200°C for 10 and 20 min, respectively.

In this research, dried acetone:chloroform extract of aerial parts of E. denticulata as one of the endemic plants to Iran, afforded a number of triterpenes and steroids including: betulin, 24-methylene-cycloart-3-ol, cycloart-23Z-ene-3β,25-diol, cycloart-23Z-ene-3β,25-diol, ergosta-8,24-dien-3-ol (obtusifoliol) and beta-sitosterol which were reported for the first time from this plant. The structure of these compounds was elucidated by NMR and mass spectroscopic methods. The MTS assay was used to determine the toxicity and antiviral activity of betulin and (3β,23E)-Cycloarta-23-ene-3,25-diol. Betulin showed anti-HSV-1 activity with EC50 value of 84.37±0.02 µg/ml, and toxicity on normal vero cells with CC50 value of 660.718±0.072 µg/ml. (3β,23E)-Cycloarta-23-ene-3,25-diol showed antiviral effect with EC50 value of 86.63±0.03 µg/ml, and toxicity with CC50 value of 1089.21±0.25 µg/ml. The results revealed that these two compounds have the antiviral activity far below the CC50 value with selectivity index (CC50/EC50) values of 7.83, and 12.57, respectively.

Ipomoea reniformis Roxb. (Convolvulaceae) is a small, weedy herb used for the management of cardiac problems in traditional systems of medicine in India and Pakistan. Objective of the present study is to investigate the hypotensive, diuretic and angiotensin converting enzyme (ACE) inhibitory activities of the aqueous-methanol (30:70) crude extract of the dried aerial parts of I. reniformis (Ir.Cr.) in rats. To record blood pressure lowering effects of the Ir.Cr., different doses of the extract were administered through jugular vein to the ketamine-diazepam anesthetized normotensive rats and blood pressure was recorded via carotid artery. ACE inhibitory activity of the extract was studied in vitro; using hippuryl-l-histidyl-l-leucine as substrate and the product hippurate was quantified spectrophotometrically after reacting with cyanuric chloride/dioxane reagent. Effects of intraperitoneal administration of the extract on urine and urinary electrolyte excretion were also investigated in rats.The extract (Ir.Cr.) produced 21.51 ± 3.41, 28.99 ± 2.30, 53.34 ± 0.88 and 61.71 ± 3.37% fall in mean arterial blood pressure of the anesthetized rats at the doses of 0.1, 0.3, 1.0 and 3.0 mg/Kg, respectively. Ir.Cr. was found to have serum ACE inhibitory activity, with IC50 value of 422 ± 21.16 μg/ml. The extract also increased urine volume and urinary Na+ excretion significantly at the doses of 30 and 50 mg/Kg in rats. The study concludes that the crude extract of Ipomoea reniformis (Ir.Cr.) has hypotensive, ACE inhibitory and diuretic activities, which provide the scientific justification for the traditional uses of the plant as cardioprotective, antihypertensive and diuretic remedy.

The rhizome of Anemarrhena asphodeloides is used as food and traditional Chinese medicine for its hypoglycemic effect. The aim of this study was to investigate the isolation, purification and hypoglycemic activity of Anemaran as the active component. The influence factors (isolation duration, ratio of residuals to water and extracting times) during the isolation process were evaluated. The optimal conditions for NA and AA were extraction temperature 90ºC and 100ºC, duration 1h and 1.5 h, extraction time 3 and 3, and the solid–liquor ratio 1:20 and 1:15, respectively. Neutral and acid Anemaran (NA and AA) were isolated from the rhizome of Anemarrhena asphodeloides. Five fractions of NA-1, NA-2, NA-3, AA-1 and AA-2 were obtained after crude neutral and acid Anemaran purified through DEAE-52 cellulose anion-exchange column. The characterizations of Anemaran and its different fractions were both analyzed by Fourier transform infrared spectroscopy (FT-IR) and scanning electron micrographs (SEM). Structural properties of different fractions were examined by FT-IR. Strong characteristic absorption peaks were observed at around 1744 cm−1and 1650 cm−1 caused by the C=O group of uronic acids, and the band between 1440 cm−1 and 1395 cm−1 associated with the stretching vibration of C–O of galacturonic acid. Neither the crude neutral, nor the acid anemaran significantly inhibited the growth of HepG2 cells in-vitro, which indicated the low cytotoxicity of the anemaran. Furthermore, both neutral and acid anemaran showed hypoglycemic effect. The hypoglycemic effect of neutral anemaran was much higher than that of acid anemaran.

Fragrant species of the genus Salvia have been attributed many medicinal properties, which include anticancer activity. In the present study, cytotoxic properties of total methanol extract of Salvia chloroleuca Rech. f. & Aellen and its fractions were investigated on MCF-7, a breast carcinoma cell line. Malignant and non-malignant cells were cultured in RPMI medium and incubated with different concentrations of plant extracts. Cell viability was quantitated by 3-(4,5-dimethylthiazol-2-yl) -5-(3-carboxymethoxyphenyl) -2-(4-sulphophenyl) -2H-tetrazolium (MTS) assay. Apoptotic cells were determined using propidium iodide (PI) staining of DNA fragmentation by flow cytometry (sub-G1 peak). S. chloroleuca inhibited the growth of malignant cells in a dose-dependent manner. Among solvent fractions of S. chloroleuca, the n-hexane and methylene chloride fractions were found to be more toxic compared to other fractions. S. chloroleuca-induced a sub-G1 peak in flow cytometry histogram of treated cells compared to control and DNA fragmentation suggested the induction of apoptosis. Administration of N-acetyl cysteine and vitamin C two ROS scavengers also resulted in significant inhibition of cytotoxicity induced by S. chloroleuca. These results support a mechanism whereby S. chloroleuca induces apoptosis of MCF-7 human breast cells through a ROS-mediated pathway.

The plants of the genus Salvia synthesize several types of secondary metabolites with antimicrobial, cytotoxic, and radical scavenging activities and are used in the folk medicine of different countries. Eleven Salvia species including S. aegyptiaca, S. aethiopis, S. atropatana, S. eremophila, S. hypoleuca, S. limbata, S. nemorosa, S. santolinifolia, S. sclarea, S. syriaca, and S. xanthocheila were collected from different localities in Iran and screened for their cytotoxic activity using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay. The antioxidant potential and total phenol contents of the plant extracts were assessed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay and Folin-Ciocalteu reagent respectively and finally antimicrobial activity of the these extracts were determined by using agar disc diffusion (ADD) and nutrient broth micro-dilution (NBMD) bioassays. Cytotoxic activity of methanol, 80% methanol and dichloromethane extracts of these plants were assessed on 3 human cancer cell lines. All of the extracts of S. eremophila and S. santolinifolia were active at IC50 values of 10.5-75.2 µg extract/mL, while the methanol and dichloromethane extracts of S. limbata, S. hypoleuca and S. aethiopis showed considerable cytotoxic activity against the tested cell lines. Among the tested plants for their antioxidant activity, S. nemorosa, S. atropatana, S. santolinifolia, and S. eremophila were the most active radical scavengers with higher total phenol contents while S. limbata, S. xanthocheila and S. aegyptiaca were the weakest ones respectively. The methanol extracts of S. santolinifolia, S. eremophila, S. sclarea and S. limbata inhibited the growth of all tested bacterial strains.

Rheum turkestanicum Janischew. (Polygonaceae) is a plant that grows in central Asia and in north-east of Iran. Traditionally, people use roots of R. turkestanicum as an anti-diabetic and anti-hypertensive as well as anticancer agent. In this study the cytotoxicity and apoptogenic properties of ethyl acetate (EtOAc), n-hexane and H2O extracts from Rheum turkestanicum Janischew. (Polygonaceae) root were determined against HeLa and MCF-7 cell lines and human blood lymphocytes. Malignant and non-malignant cells were cultured in RPMI 1640 medium and incubated with different concentrations of plant extracts. Cell viability was measured by MTS assay. Apoptotic cells were evaluated using PI staining of DNA fragmentation by flow cytometry (sub-G1 peak). The degree of DNA fragmentation was analyzed using agarose gel electrophoresis based on the formation of inter-nucleosomal units. The expression of apoptosis-related protein Bax and PARP cleavage were detected by Western blotting. EtOAc and n-hexane extracts decreased cell viability in malignant but not in non-malignant cells, as a concentration and time dependent manner. EtOAc extract induced a sub-G1 peak in flow cytometry histogram of treated cells compared to the control. DNA fragmentation indicating apoptotic cell death was involved in R. turkestanicum induced toxicity and cleaved PARP fragment was also detected. In conclusion, this is the first report on the cytotoxic effects of R. turkestanicum in which apoptosis played an important role. However, further evaluations are needed to fully understand the possible anti-tumor properties.

Diazinon is an organophosphate which is extensively used in trade and agriculture. Due to its widespread application, its toxicity is common. Several studies have shown that organophosphates are able to induce oxidative stress by generating free radicals and depletion of endogenous antioxidants. Pomegranate seed oil (PSO) possesses anti-inflammatory and antioxidant effects. In this study, the effect of PSO was evaluated on diazinon-induced nephrotoxicity in rat. Wistar male rats were randomly divided into four groups, 6 each. Group one received saline, 1 mL/kg, group 2 received diazinon 100 mg/kg. Groups 3 and 4 received PSO, 0.32 and 0.64 mg/kg, one hour before diazinon 100 mg/kg respectively. After 24 h, animals were anesthetized. Blood samples were taken out by cardiac puncture for measuring the level of serum urea and creatinine. 24 h urine samples were also collected for measuring glucose and protein concentration. The right kidney was removed and homogenized for measuring malondialdehyde and thiol content Compare to control group, DIZ increased urea and serum creatinine, urinary glucose, and malondialdehyde, but did not modify significantly urinary protein and thiol content. In groups received PSO+ DIZ, serum creatinine, urinary glucose and MDA were significantly decreased. DIZ induced acute nephrotoxicity and oxidative stress. Probably, increasing of serum creatinine and urinary glucose are appropriate markers for diagnosis of kidney damage. In addition increasing of MDA level emphasizes that DIZ plays role in pathogenesis of kidney via oxidative stress mechanism. PSO reduced DIZ toxicity by antioxidant activity.

In this research, we investigated the cytotoxic mechanisms of Cochlodinium polykrikoidescell lysate on isolated rat liver hepatocytes.This micro algae is responsible for a severe and widespread harmful algal bloom in the Persian Gulf and Gulf of Oman (2008-2009). Isolated hepatocytes were obtained by collagenase perfusion of Sprague-Dawley rat liver.According to our results, incubation of algal lysate with isolated rat hepatocytescaused hepatocyte membrane lysis, reactive oxygen species (ROS) formation, glutathione depletion, collapse of mitochondrial membrane potential,ATP depletion and increase in ADP/ATP ratio, cytochrome c release in to the hepatocyte cytosol,activation of caspase-3 (final mediator of apoptosis) and appearance of apoptosis phenotype. On the other hand, pretreatment of antioxidants (α-tocopherol succinate and BHT), hydroxyl radical scavengers (mannitol and DMSO), mitochondrial permeability transition (MPT) pore sealing agents (cyclosporine A, carnitine and trifluoperazine), NADPH P450 reductase inhibitor (Diphenyliodonium chloride), CYP2E1 inhibitors (Phenylimidazole and 4-Methylpyrazole) andATP generators (L-glutamine, Fructose and Xylitol)inhibited caspase-3 activation and cell death in algal lysate treated hepatocytes.Our data also confirmed that algal lysate activates apoptosis signaling via oxidative stress and mitochondrial pathway. Thus, ROS formation caused by the lysate exposure could directly be involved in mitochondrial MPT pore opening and activation of caspase-3 leading to C.polykrikoides lysateinduced apoptosis on rat hepatocytes. These findings contribute to a better understanding of C.polykrikoides-toxic effects on mammalian liver cells.

Methylsulfonylmethane (MSM) is a sulfur-containing compound commonly found in diet and known to reduce oxidative stress. This trial was conducted to determine whether single dose supplementation with MSM attenuates post-exercise oxidative stress in healthy untrained young men. Sixteen untrained men volunteered for this study. Participants were randomized in a double-blind placebo-controlled fashion into 2 groups: Methylsulfonylmethane (MSM) (n = 8) and placebo (n = 8). The participants took supplementation or placebo before running on treadmill for 45 min at 75% VO2max. The MSM supplementation was prepared in water as 100 mg/kg body weight. The placebo group received water. Serum Malondealdehyde (MDA), uric acid, bilirubin, protein carbonyl (PC) and plasma vitamin E levels were determined as the markers of oxidative stress. Plasma GSH (reduced Glutathione) and total antioxidant capacity (TAC) were measured as markers of plasma antioxidant system. MSM supplementation successfully lowered serum PC 2 and 24 h after exercise. Plasma TAC in MSM group was higher at 24 h after exercise. Serum level of uric acid and bilirubin were significantly low immediately after exercise in MSM supplemented group. There was no significant difference between groups in terms of plasma GSH level. These results complement earlier studies showing anti-oxidant effect of MSM and suggest that single dose oral supplementation with MSM lowers exercise induced oxidative stress in healthy untrained young men, but is not adequate to significantly affect plasma GSH level.

The goal of this study was to determine the effects of the L-type calcium channel blockers verapamil and diltiazem on the currents of voltage-gated potassium channel (fKv1.4ΔN), an N-terminal-deleted mutant of the ferret Kv1.4 potassium channel. Measurements were made using a two electrode voltage clamp technique with channels expressed stably in Xenopus oocytes. The fKv1.4ΔN currents displayed slow inactivation, with a half-inactivation potential of –38.38 mV and slow recovery from inactivation (τ = 1.90 seconds at –90 mV). The fKv1.4ΔN currents exhibited state-dependent blockade by both drugs, and the inhibition was frequency-, voltage-, and concentration-dependent, consistent with open channel block. Verapamil and diltiazem blocked fKv1.4ΔN currents with 50% inhibitory concentration (IC50) values of 260.71 ± 18.50 μmol/L and 241.04 ± 23.06 μmol/L, respectively. Verapamil accelerated the C-type inactivation rate and slowed recovery of the fKv1.4Δ N channel, while shifting the steady activation curve to the right. Blockade of fKv1.4ΔN currents by diltiazem was similar to that of verapamil, but diltiazem accelerated the decay rate of inactivation of fKv1.4ΔN currents without modifying the kinetics of current activation. The present results suggest that verapamil and diltiazem accelerate the C-type inactivation and slow the recovery of the fKv1.4ΔN channel in the open state.

In this study, we investigated the cardioprotective effects of resveratrol. Rats were intraperitoneally administered with resveratrol (25 mg/kg bw) or vehicle (ethanol 10%) for 7 days and their heart subjected to ischemia/reperfusion injury. Isolated hearts were langendorff perfused, left ventricular functions as heart rate and developed pressure, as well as, heart antioxidant status were determined. Data showed that resveratrol improved recovery of post-ischemic ventricular functions when compared to control. Resveratrol also improved myocardial lipoperoxidation, free iron and antioxidant enzyme activities. Resveratrol decreased significantly catalase while it increased peroxidase and superoxide dismutase activities. In this later case, native PAGE analysis of superoxide dismutase isoforms revealed that resveratrol up regulated iron and manganese isoforms. Resveratrol exerted potential cardioprotection partly by its antioxidant properties.

Several studies point to an important function of cyclooxygenase (COX) and prostaglandin signaling in models of synaptic plasticity which is associated with N-methyl-D-aspartate receptors (NMDARs). Cyclooxygenase gene is suggested to be an immediate early gene that is tightly regulated in neurons by NMDA dependent synaptic activity. Nonsteroid Antiinflammatory Drugs (NSAIDs) exert their antiinflammatory effect by the inhibion of COX have controversial effects on learning and memory. We administered ibuprofen as a non-selective COX-2 inhibitor and nimesulide as a selective COX-2 inhibitor for 8 weeks for determining the cognitive impact of subchronic administration of NSAIDs to aged rats. Wistar albino rats (16 mo, n = 30) were separated into control (n = 10), ibuprofen (n = 10) and nimesulide (n = 10) treated groups. First we evaluated hippocampus-dependent spatial memory in the radial arm maze (RAM) and than we evaluated the expression of the NMDAR subunits, NR2A and NR2B by western blotting to see if their expressions are effected by subchronic administration with these drugs.Ibuprofen and nimesulide treated rats completed the task in a statistically significant shorter time when compared with control group (p < 0.01), but there was no statistically significant difference between groups about choice accuracy data in RAM. Furthermore, no statistically significant difference was detected for the protein expressions of NR2A and NR2B of the subjects. Oral administration of ibuprofen and nimesulide for 8 weeks showed no impairment but partly improved spatial memory.

Paraquat is a commonly used herbicide in many countries which can lead to systematic poisoning on exposure, In this study, paraquat (PQ)-induced changes in the expression of Cyclooxygenase-2 (COX-2) along with biochemical and histopathological changes in the lungs, liver and kidneys were studied. Twenty four male Wistar rats (180-200 g) were exposed either against saline as control group or various doses of PQ (3.5, 7 and 10 mg/kg, SC) as test groups for 7 consecutive days. The animals in test groups demonstrated a significant increase of malondialdehyde and NO contents, while a remarkable decrease of total thiol molecules was recorded. Histopathological studies revealed a severe alveolar edema and hemorrhages in the lungs, congestion and glycogen degeneration in the liver and multifocal interstitial nephritis along with proximal tubular degeneration in the kidneys. PQ up-regulated the COX-2 expression at mRNA level significantly in the examined organs. This data suggest that the PQ-induced oxidative disturbances and pathological damages can be attributed to the upregulation of COX-2 in examined organs.

Diazinon (DZN) is a synthetic organophosphorus (OPs) insecticide widely used in agricultural and household applications. OPs، particularly DZN، are highly lipid soluble liquids. Intravenous lipid emulsion (ILE) has been shown to reduce toxicity caused by some lipid soluble agents. We evaluated the antidote effect of ILE on acute toxicity of DZN. Twenty-four Sprague-Dawley female rats weighting 200-250 g were treated orally with dose of 480 mg/kg of DZN gavaged at the volume of 0. 5 mL/kg. Thirty minutes after administration of DZN، two groups were treated by either ILE 10% (ILE10) or normal saline (NS) (16 mL/kg) (NS16) that were infused for the duration of 15 minutes. Another two groups were also treated by either ILE 20% (ILE20) or NS (10 mL/kg: NS10) as above. The changes in body weight، diarrhea score، muscular power، fasciculation، convulsions and mortality rate of the animals were all monitored immediately after infusions and then every 6 h up to 48 h. There was no significant difference in animals mean weight between different groups during the observation period. In addition، during the 48-hour observation we could not find any difference in muscular power and diarrhea score between groups of ILE20-NS10 and ILE10-NS16 in comparison with each other، and neither ILE 10% nor ILE %20 could not reduce mortality rate of animals or increase the survival time of rats. In conclusion،ILE seems to be unable to reverse DZN acute toxicity and it might be due to conversion of DZN to potent and less lipid soluble agent.

Artemisinin and its derivatives are very important new class of antimalarial drugs. One of the most important artemisinin derivatives is artemether. The antiparasitic activity of artemether as a derivative of artemisinin is related to endoperoxide bridge in its structure. The aim of this study was the evaluation of antileishmanial effect of artemether, with more focus on its apoptotic effect. In this study we used artemether in concentration of 5, 10, 25, 50 and 100 μg/ml for promastigote assay, promastigote proliferation measurements by MTT assay, detection of apoptotic cells by Flow cytometry analysis and DNA ladder assay. The application of artemether, promastigote IC50 was measured as 25 μg/ml. The percentage of apoptotic promastigotes by using 25 μg/ml of artemether was 42.28. The results of present study showed that artemether has inhibition effect on intracellular and extracellular growth of Leishmania major. Promastigotes of Leishmania major undergo apoptosis after exposure to artemether.

Glutathione (GSH) is one of the most important antioxidants that plays an essential role in detoxification of reactive oxygen species (ROS) which oxidizes to glutathione disulfide (GSSG). Paraquat (PQ), awidely used herbicide, causes pulmonary injury with the productionof ROS. Excessive ROS accumulation as a consequence of PQ exposure are frequently targeted by GSH thereby oxidative stress leads to depletion of cellular GSH by transforming of GSH to glutathione disulfide (GSSG). A precise method of measuring of GSSG concentration in plasma as indicator of oxidative stress is needed. Some analytical techniques such as high-performance liquid chromatography (HPLC), gas chromatography and capillary electrophoresis have been used for determination of GSSG concentration. In the present study, a new HPLC method with fluorescence detection based on derivatization of the amine group of glutathione with 9-fluorenylmethyl chloroformate (FMOC-Cl) was developed. Male Wistar albino rats exposed to different doses of PQ (20-60 mg/kg) and control group were used and after protein precipitation, their plasma was subjected to derivatization with FMOC in the presence of borate buffer. The derivatized samples were injected to HPLC system with C18 column, mobile phase consisting of methanol and phosphate buffer, λem= 315 nm, λex = 260 nm. Among all experimental groups, the rats which received 60 mg/kg PQ, showed a significant increase in the amount of oxidized glutathione (GSSG) compared to the control group. In this study, the applied derivatization and HPLC method made it possible to measure small amounts of glutathione in plasma using a precise and sensitive technique.

Effects of inoculation level (4 or 8-fold compared to standard inoculation) and order (standard inoculation before fermentation and 3-fold inoculation at the end of fermentation = 1+3, Two-fold inoculation before fermentation and the same at the end of fermentation = 2+2, 2+6, 4-fold before fermentation = 4, 4+4, and 8) of culture inoculums containing probiotics on viscosity, phase separation, particle size analysis, microstructure and sensory attributes of probiotic Doogh were studied. The probiotic microorganisms were Lactobacillus acidophilus LA-5 and Bifidobacterium lactis BB-12. Treatments with 2- and 4-fold inoculation before fermentation had the highest instrumental viscosity and surface tension at the end of fermentation. The size diameter of particles in the structure of treatment 8I was significantly (p < 0.05) smaller than I after stirring with a Lab stirrer (1500 rpm), and even after homogenization with a homogenizer (150 bar). 8I was an un-uniform, disintegrated and clumped structure with limited junctions in its network that resulted in a weak structure with bigger particles after agitation and smaller particles after stirring and homogenization compared to other treatments. This treatment also had the lowest record in ranking sensory test among treatments with a mixed culture-like and vinegary-like taint. Overall, treatments with 2- and 4-fold inoculation were realized as the best from the sum of physiochemical and sensory properties point of view.

MarA activates two membrane dependent mechanisms of resistance to different antibiotics, such as ciprofloxacin and tetracycline, including promotion of outflux and inhibition of influx of antibiotics. Thus, MarA causes multiple antibiotic resistance phenotype. The activation of these mechanisms needs overexpression of marA. This could happen through mutation in marR. Thus, the aim of this study was to measure marA expression in ciprofloxacin resistant E. coli gyrA mutants and clones with or without marR mutation. For this purpose, real time PCR was used to measure relative expression of marA in above mutants and clones. Results showed that two clones, C14 and C17 overexpressed marA. It is concluded that the level of marA expression is important for activation of above mechanisms.

Inflammatory bowel disease (IBD) is an irregular response of immune system accompanied with different inflammatory manifestations including alterations in cytokines. Probiotics are non-pathogenic organisms with probable effects in various conditions such as inflammation. The present study hypothesized whether oral intake of bifidobacterium and lactobacillus in form of probiotic yogurt may represent an immunomodulatory effect in IBD patients. Overally, 210 patients in remission phase and 95 healthy people were recruited. Patients were randomly divided into two groups of either 250 grams of probiotic yogurt (PI) or 250 grams of plain yogurt (PC) daily for 8 weeks. The healthy control group (HG) also received probiotic yogurt as noted. The serum levels ofcytokines TNF-α, IL-6, IL-1β, IL-10 and CRP levels were measured at baseline and at termination time. A significant difference was observed between intervention groups of PI and PC with HG group (p<0.05). After the intervention, serum levels of IL-1β, TNF-α and CRP were significantly decreased in PI group compared to their baseline values and intervention groups. The serum levels of IL-6 and IL-10 increased significantly after the intervention compared to baseline values and PC levels (all p values<0.05). Intestinal homeostasis is a balance between pro and anti-inflammatory responses of intestinal immunocytes and could be maintained by probiotics.

Medication errors have important effects on increased length of hospitalization, increased mortality and costs. We assessed the incidence of medication errors and characterize the error types in an emergency department in a large teaching hospital in Tehran. We also investigated the effect of Emergency Department pharmacists on patient safety with regard to recovery of potentially harmful medication errors. The study was conducted in the 24 bed emergency department from February to March, 2010 at a 600-bed teaching hospital. Two hospital pharmacists and two clinical pharmacy residents observed care provision and collected data on medication errors. Demographic data, type of medication error, the recorded stage of error, date and time of occurrence and report, who made the error, probability of error were recorded from medical records. We used chi-squared and independent sample t- tests for analyzing the data. We recorded 203 medication errors during 180 hours. The incidence of medication errors was 50.5% at various levels in the emergency department. Significant difference in age means was seen between the patients with and without medication errors. Seventy four point nine percent of errors were recorded as definitely an error. Most recorded errors were made by nurses (44.5%) and occurred in administrating stage (63.6%). Given that the rate of the errors was relatively high, it seems that the presence of clinical pharmacist can be beneficial.

Atrial fibrillation (AF) is associated with inflammatory and hypercoagulability state. Previous studies evaluated the safety and efficacy of dabigatran and warfarin in prevention of thrombothic complications. This study was intended to assess the influence of these drugs on hemostatic and inflammatory markers among patient underwent pulmonary vein ablation. A total of 100 patients with AF who underwent catheter ablation were randomized to treatment with dabigatran (D) 110 mg twice daily or warfarin (W) adjusted to an international normalized ratio (INR) of 2.0 to 3.0 for 3 months after ablation procedure. C - reactive protein (CRP), D-dimer, prothrombin fragment F1 + 2 (F1 + 2), were measured at baseline before ablation procedures, after 30 days and after 90 days of treatment. After 3 months, the D-dimer was 164.9 ± 48.9 in Dabigatran and 197.2 ± 58.6 in warfarin group, F1 + 2 was 0.4 ± 0.2 in dabigatran and 0.8 ± 0.2 in warfarin group and CRP level was 1.8 ± 1.6 in Dabigatran and 5.1 ± 5 in warfarin group. (All p-values < 0.05) The results showed that treatment with dabigatran made greater changes in the serum level of CRP, D-dimer, F1 + 2. The pattern of changes in serum CRP levels D-dimer, F1 + 2 is much faster and with a greater slope in the dabigatran group.

Pharmacists are members of the healthcare teams that provide valuable services to society. Their incentive to deliver such services is influenced by remuneration methods. In this study, we aimed to review the remuneration models for pharmacists’ services and the factors affecting the profitability of pharmacies in some selected countries, including France, Ireland, Canada and Turkey, and compared them to Iran. International data were collected by literature review on Google, Google scholar, PubMed and Scopus. In addition, domestic data were collected by contacting relevant organizations. There is no payment for pharmacists’ cognitive services in Iran and in the countries investigated, except for some Canadian provinces. The dispensing fee system in Iran does not seem to be adequate, especially considering that most of the insurers do not cover these fees. The pricing method in Iran has resulted in a low price level, in comparison to the other countries, and this issue has dramatically affected the profitability of pharmacies in standard practice. It could be concluded that changing the current formulation for the dispensing fee to a more appropriate one, defining a remuneration system for non-owner pharmacists other than salary and implementing the new pricing method are necessary in order to improve the services provided by pharmacies.