DARU, Journal of Pharmaceutical Sciences
Volume:17 Issue: 3, Autumn 2009

Background and the purpose of the study: Of the gene delivery systems, non-viral polycationic gene delivery nanosystems have been alternatively exploited as a relatively safe delivery reagents compared to viral vectors. However, little is known about the genomic impacts of these delivery systems in target cells/tissues. In this study, the toxicogenomics and genotoxicity potential of some selected polycationic lipid/polymer based nanostructures (i.e., Oligofectamine® (OF), starburst polyamidoamine Polyfect® (PF) and diaminobutane (DAB) dendrimers) were investigated in human alveolar epithelial A549 cells. To study the nature and the ontology of the gene expression changes in A549 cells upon treatment with polycationic nanostructures, MTT assay and microarray gene expression profiling methodology were employed. For microarray analysis, cyanine (Cy3/Cy5) labeled cDNA samples from treated and untreated cells were hybridized on target arrays housing 200 genes.Results and major The polycationic nanosystems induced significant gene expression changes belonging to different genomic ontologies such as cell defence and apoptosis pathways. These data suggest that polycationic nanosystems can elicit multiple gene expression changes in A549 cells upon their chemical structures and interactions with cellular/subcellular components. Such impacts may interfere with the main goals of the desired genemedicine.

A new herbal drug, Setarud (IMODTM) that has been shown to have beneficial immune effects was tested to determine its acute and chronic toxicity in animals and to establish its intravenous form maximum tolerated dose (MTD) in an open-labeled phase I clinical trial. BALB/c and C57BL/6 mice and Wistar rats were monitored for general state and biochemical markers for chronic test. At the end of chronic test, animals examined macroscopically and histologically. HIV-infected asymptomatic male patients with CD4 counts more than 200, were enrolled in the trial. Baseline dose was calculated from the 10% lethal dose (LD10) established in laboratory animal studies. Dose escalation was performed in four cohorts of 3 patients receiving IMODTM intravenously at a cohort-specific dose of 2, 4, 6.7, and 10 ml daily for 4 weeks. Patients were clinically examined at days of 1, 2, 3 and then weekly; and the safety was assessed on the basis of reports of adverse events, laboratory-test data and toxicity signs. LD50 values in acute toxicity test were 42-66 and 50-56 ml/kg in i.m. and i.p. injections, respectively. Total scores of embryotoxicity during pregnancy were significantly lower in the Setarud group (p < 0.05). Pre-implantational deaths in the Setarud group were significantly higher, but post-implantational deaths level was lower than those in the control group. Inhibition of ossification in the skeletons of the fetuses and incidence of still birth were significantly higher while body weight of new-born rats of treatment group in the first month of their lifes were lower than those of the control group.In all four cohorts, there were no major side effects or dose-limiting toxicity, except for a mild sweating and weight loss in two patients from the first group that was reversed without discontinuation of the treatment. No clinically relevant trends in laboratory test results or ECG changes were noted. No adverse effect due to IMODTM was observed at one month of follow-up. Maximum tolerated dose of IMODTM was 10 ml a day. Results of this study has identified a safe dose of IMODTM that can be used in future clinical trials.

Background and Purpose of the study: Hyperglycemia and insulin resistance are common findings among critically ill patients. Intensive insulin therapy reduces morbidity and mortality in patients of surgical and medical intensive care units (ICUs), but its role in patients of general intensive care units still remains unknown. The present study was designed to determine the effect of intensive insulin therapy on ICU mortality.] Adult patients admitted to general intensive care units in Valiy-e- Asr Hospital, who required intensive care for at least five days were considered for a prospective, randomized and control study. On admission, patients were randomly chosen either to normalize their blood glucose levels or to prepare them for conventional therapy. Intensive insulin therapy reduced blood glucose levels of 129 patients but did not have any significant effect on reduction of mortality rate of hospitalized patients (30.6% in the conventional-treatment group vs. 38.8% in the intensive-treatment group, p > 0.05). However, the morbidity rate was significantly plummeted as a consequence of acceleration in the process of weaning of the patient from mechanical ventilation, and subsequently discharging from the ICU. The benefit of intensive insulin therapy was attributed to its effect on mortality among patients who remained in the intensive care unit for more than five days (78.9% conventional-treatment group vs. 46.2% intensive-treatment group, p = 0.04). Intensive insulin therapy significantly reduced morbidity but not mortality among the patients in general intensive care units. The very possible risk of subsequent disease-associated and fatal complications was reduced in patients who were treated for five days or longer time. Further studies are required to confirm these preliminary results.

Background and the purpose of the study: Tuberculosis is a curable disease if diagnosed and treated properly with anti-tuberculosis drugs. These drugs can cause severe adverse reactions including hepatotoxicity.The goal of this study was to evaluate the rate and the time of incidence, pattern of alterations in liver enzyme, risk factors and outcome of anti-tuberculosis drugs induced hepatotoxicity in Iranian Tuberculosis patients. In a prospective cohort study, 102 patients (68 male, 34 female, mean age 43.21±18 years) with tuberculosis diagnosis were followed during anti-tuberculosis drug treatment course. Drug related hepatotoxicity was defined as increase in serum alanine aminotransfrase or aspartate aminotransfrase greater than three or five times of the upper limit of normal, with or without symptoms of hepatitis, respectively. anti-tuberculosis induced hepatotoxicity was detected in 32 (31.37%) of the patients. Human immunodeficiency virus and hepatitis C virus infections, concomitant use of hepatotoxic drugs, and abnormal baseline serum alanine aminotransfrase and aspartate aminotransfrase level were risk factors for anti-tuberculosis drugs induced hepatotoxicity. Anti-tuberculosis drugs induced hepatotoxicity is a major problem in Iranian tuberculosis patients and cause treatment interruption in 31.37% of patients.

The plant Pergularia daemia (Asclepiadaceae) and Carissa carandas (Apocynaceae) are traditionally used as a medicinal agent and they are widely distributed to tropical and subtropical region of India. In the present study the folklore uses of P. daemia and C. carandas was investigated. The analgesic activity was studied in mice using hot plate and acetic acid induced writhing methods, while carrageenan induced paw edema was used to access anti-inflammatory activity. The antipyretic activity was evaluated by Brewer''s yeast induced pyrexia in rats. The ethanol and aqueous extracts from roots of P. daemia and C. carandas exhibited significant (p < 0.01) analgesic, anti-inflammatory and antipyretic activities at the doses of 100 and 200 mg/kg body weight. In analgesic activity, the highest reaction time was observed (9.8 sec.) from ethanol extracts of P.daemia at a dose of 200 mg/kg body weight, while highest percentage of inhibition of abdominal constriction (72.67%) was observed for ethanol extracts of C.carandas at a dose of 100 mg/kg body weight. The ethanol and aqueous extracts of P. daemia and C. carandas were found to reduce significantly the formation of edema induced by carrageenan after 2 hrs. Both plants showed significantly competent on yeast induced hyperpyrexia in rats after 2 hrs. The results of this study indicated that the ethanol and aqueous extracts from roots of P. daemia and C. carandas possess significant analgesic, anti-inflammatory and antipyretic activities in rodent models.

Background and the purpose of study: Rosa damascena Mill. (Rosaceae) has cooling, soothing, astringent, and anti-inflammatory effects, and has been used in the north of Iran as a cardiotonic agent. The aim of this study was to identify components of R. damascena (cultivated in Guilan Province) extract and essential oil and to study their biological activities. Essential oil of R. damascena was prepared by hydrodistillation and analyzed with GC/MS instrument. The antioxidant activity of hydro-alcohlic extract of petals and essential oil was measured using free radical scavenging activity with 2-2-diphenyl, 1-picrylhydrazyl (DPPH) and lipid peroxidation (ferric ammonium thiocyanate) methods. Hydro-alcoholic extract showed strong free radical scavenging capacity compared to lipid peroxidation inhibitory effects. IC50 values of the extract were 2.24 μg/mL and 520 μg/mL in free radical scavenging and lipid peroxidation assays, respectively. The major components of essential oil were linalool (3.8%), nerol (3.05%), geraniol (15.05%), 1-nonadecene (18.56%), n-tricosane (16.68%), hexatriacontane (24.6%) and n-pentacosane (3.37%). The bioassay-guided fractionation of extract led to the isolation of three flavonol glycosides: quercetin-3-O-glucoside, kaempferol-3-O-rhamnoside and kaempferol-3-O-arabinoside. The IC 50 value of the radical scavenging activity of kaempferol-3-O-rhamnoside which was, 530 μg/mL was weaker than the extract.Major The petal of this cultivated rose has no bitter taste and because of its potential antioxidant activity and good taste, can be used as food flavor and a preventing agent for many diseases.

Background and purpose of the study:Screening of different plant components for new anticancer drugs is one of the main research activities throughout the world. In this study, the anticancer effects of Astrodaucus persicus, an Iranian species of family of Umbelliferae, in human breast cancer T47D cells was investigated. Also since tumorigenesis is thought to result from a series of progressive gene alterations, including activa tion of oncogenes and inactivation of tumor suppres sor genes, expression of two such genes, p53 and Bcl-2 that are believed to play a crucial role in tumorigenesis and cell death were determined. The p53 and Bcl-2 genes and proteins expression alterations in T47D cells at RNA synthesis level was studied by using RT-PCR analysis and protein synthesis using immunocytochemistry technique. p53 gene expression increased significantly in the presence of both plant extracts but Bcl-2 expression increased significantly in the presence of aerial and decreased significantly in the presence of root extract. In addition, treatment of T47D cells with plant extracts decreased the nuclear staining of p53 and cytoplasmic staining of Bcl-2 proteins. These results suggest that the methanolic extracts of Astrodaucus persicus especially its root extract may contains bioactive compounds, probably coumarins that prevents proliferation of T47D breast carcinoma cells by mechanisms such as apoptosis. These data are the first report on the possible molecular mechanisms of action of Astrodaucus persicus extracts in breast cancer cell proliferation.

Prolonged storage of cryopreserved isolated hepatocytes is now possible and such cells have been used with success for drug metabolism and toxicity studies. Quality of the cells before cryopreservation is important for viability and function after freezing. In this study, fresh rat hepatocytes were incubated with the thiol-containing compounds N-acetylcysteine (NAC), dithiotheritol (DTT) and fructose as ATP supplier, at incubation times 1 and 3 hrs. The preincubated hepatocytes cryopreserved for 24 hour, and 1 and 3 months. Hepatocytes, viability were determined immediately postthaw by Trypan Blue exclusion. Fructose preincubation improved the viability of hepatocytes at a concentration of 300 mM. Preincubation with DTT (50, 100 and 200 μM) prior to cryopreservation had beneficial effects on viability of hepatocytes. The postthaw viability of hepatocytes preincubated with NAC was uniformly poor. Preincubation with DTT, improved GSH levels before freezing which could be responsible for the reduction in membrane damage during cryopreservation. It may be concluded that the intracellular

Background and the purpose of the study: Taxol, a natural antitumor agent, was first isolated from the extract of the bark of Taxus brevifolia Nutt., which is potentially a limited source for Taxol. In the search of an alternative source, optimum and cost benefit extracting solvents, various solvents with different percentage were utilized to extract Taxol from needles of Taxus baccata. One g of the dried needles of Taxus baccata, collected from Torkaman and Noor cities of Iran, was extracted with pure ethanol or acetone and 50% and 20% of ethanol or acetone in water. Solvents were evaporated to dryness and the residues were dissolved in 5 ml of methanol and filtered. To one ml of the filtrate was added 50 μl of cinamyl acetate as the internal standard and 20 μl of the resulting solution was subjected to the HPLC to determine the extraction efficiencies of tested solvents. Five μl of filtrate was also subjected to the LC-MS using water/acetonitrile (10/90) as mobile phase and applying positive electrospray ionization (ESI) to identify the authenticity of Taxol. Results of this study indicated that Taxol extraction efficiency was enhanced as the percentage of ethanol or acetone was increased. HPLC analysis showed that Taxol could be quantified by UV detection using standard curve. The standard curve covering the concentration ranges of 7.8 - 500 μg/ml was linear (r2= 0.9992) and CV% ranged from 0.52 to 15.36. LC-MS analysis using ESI in positive-ion mode confirmed the authenticity of Taxol (m/z 854; M+H), as well as some adduct ions such as M+Na (m/z 876), M+K (m/z 892) and M+CH3CN+H2O (m/z 913). The results suggest that 100% acetone is the best solvent for the extraction of Taxol from Taxus baccata needles.

Background and the purpose of the study: This study reports the laboratory optimization for the preparation of salbutamol sulphate-ethylcellulose microparticles by a non-solvent addition coacervation technique through adjustment of the ratio of salbutamol sulphate to ethylcellulose. The variation of drug release between the microparticles and tabletted microparticles was also investigated. In vitro release profiles of developed microparticles and tabletted microparticles were studied using USP XXIV dissolution apparatus I and II, respectively, in 450 ml double distilled water at 50 rpm maintained at 37°C. White microparticles with no definite shape having good entrapment efficiency (96.68 to 97.83%) and production yield (97.48 ± 1.21 to 98.35 ± 1.08%) were obtained. In this investigation, initial burst effect was observed in the drug release behavior. The rate of drug release from microparticles decreased as the concentration of polyisobutylene was increased from 6% to 12% during microencapsulation. The release pattern of tabletted microparticles was affected significantly (p < 0.05) by the addition of hydroxy propyl methyl cellulose (HPMC) as excepient and insignificantly (p > 0.05) by the type of dissolution media and stirring speed. Tabletted microparticles showed good stability and reproducibility. Ethylcellulose was found to be compatible with salbutamol sulphate. The drug release from all formulations was best fit to Higuchi''s equation and the mechanism of drug release was anomalous diffusion from all formulations. The results of this study suggest that by using ethylcellulose it is possible to design a single-unit, sustained-release oral dosage form of salbutamol sulphate for indication of twice a day

Background and the purpose of the study:Lecithin organogels are formed spontaneously by adding a given amount of water to lecithin/organic solvent mixture. The aim of this research was to develop and optimize a semisolid preparation with appropriate release profile. Lecithin organogels containing Propranolol hydrochloride (PR) were formulated, based on phase diagram studies, using soybean lecithin (Epikuron 200), isopropyl myristate (IPM) and propranolol hydrochloride (PR) solutions (10, 20, 30, 50 % w/w) or water at various lecithin/ IPM weight ratios. The flux and the viscosity of the prepared formulations were determined and further chosen as two responses for optimization, using experimental design and optimization methods (i.e. Modified Simplex and Central Composite Designs, respectively). Results of modified simplex runs (i.e. lecithin: 30-50%, PR: 20-40% and water: 3-4%) were also used as constraints for constructing central composite design space. The numerical and graphical optimizations were then run and the "sweet spot" corresponding to the most desirable formulation region compromising both responses were achieved. Phase diagrams showed a narrow area of existence of non-birefringent, transparent, viscoelastic region, which was extended as %PR incorporated into the system was increased. It was observed that as the lecithin concentration increased from 30 to 60 % w/w, drug incorporation capacity and viscosity increased while the flux of PR from organogels decreased remarkably. Also it was found through optimization that among the organogels investigated, those formulations containing 31.5-37.5 % w/w lecithin, 30.5-34.5 % w/w PR solutions and 3-3.35 % w/w water possessed the highest flux.Major Data confirmed that the choice of lecithin/IPM weight ratio and the amount of drug incorporated may be crucial in determining the performance of an organogel.