DARU, Journal of Pharmaceutical Sciences
Volume:17 Issue: 1, Spring 2009

Background and the purpose of the study: Ichnocarpus frutescens has been widely used in the traditional medicine for the treatment of a variety of diseases. In the present study anti-inflammatory and antioxidant property of roots of this plant was investigated. The anti-inflammatory activity of methanolic extract of Ichnocarpus frutescens (MEIF) was evaluated by carrageenan, and cotton pellet induced granuloma tests to determine its effects on acute and chronic phase of inflammation models in rats.Results and major Preliminary Phytochemical analysis of methanolic extracts showed presence of terpenoids, flavonoids, and sterols. Maximum inhibition (54.63 %) was obtained at the dose of 100 mg/kg after 3 hrs of drug treatment in carrageenan induced paw oedema, whereas indomethacin produced 57.65 % of inhibition. In the chronic model, 300 mg/kg of MEIF like indomethacin and dexamethasone standard drug decreased formation of granuloma tissue by 22.64, 29.63 % and 34.84 % respectively. The successive methanol re-extract of Ichnocarpus frutescens root (MEIF) exhibited strong scavenging effects on 2, 2-diphenyl-2-picryl hydroxyl (DPPH) free radical, nitric oxide, super oxide anion, hydroxyl radical and inhibition of lipid peroxidation. These results clearly indicate strong anti-inflammatory and antioxidant properties of Ichnocarpus frutescens root.

The ability of supercritical fluids (SCFs), such as carbon dioxide, to dissolve and expand or extract organic solvents and as result lower their solvation power, makes it possible the use of SCFs for the precipitation of solids from organic solutions. The process could be the injection of a solution of the substrate in an organic solvent into a vessel which is swept by a supercritical fluid. The aim of this study was to ascertain the feasibility of supercritical processing to prepare different particulate forms of fluticasone propionate (FP), and to evaluate the influence of different liquid solvents and precipitation temperatures on the morphology, size and crystal habit of particles. The solution of FP in organic solvents, was precipitated by supercritical carbon dioxide (SCCO2) at two pressure and temperature levels. Effects of process parameters on the physicochemical characteristics of harvested microparticles were evaluated. Particle formation was observed only at the lower selected pressure, whilst at the higher pressure, no precipitation of particles was occurred due to dissolution of FP in supercritical antisolvent. The micrographs of the produced particles showed different morphologies for FP obtained from different conditions. The results of thermal analysis of the resulted particles showed that changes in the processing conditions didn''t influence thermal behavior of the precipitated particles.Evaluation of the effect of temperature on the size distribution of particles showed that increase in the temperature from 40 oC to 50 oC, resulted in reduction of the mean particle size from about 30 µm to about 12 μm.‍ From the results of this study it may be concluded that, processing of FP by supercritical antisolvent could be an approach for production of diverse forms of the drug and drastic changes in the physical characteristics of microparticles could be achieved by changing the type of solvent and temperature of operation.

In this study antimicrobial effectiveness test was performed on eye-drops which had high microbial contaminations in hospital practice to find out whether their antimicrobial efficacies affect the magnitude of microbial contamination during their uses. Artificial tear, atropine sulfate, betamethasone, homatropine hydrobromide, phenylephrine hydrochloride, phenylephrine zinc, pilocarpine hydrochloride, tetracaine hydrochloride and tropicamide eye-drops were subjected to the United States Pharmacopeia (USP) and British Pharmacopeia (BP) antimicrobial preservative effectiveness tests. The results of this study showed that eight out of the nine products met the BP ''B'' and USP criteria. The preservative employed in phenylephrine zinc eye-drop did not possess adequate antimicrobial activity against P. aeruginosa. Other eye-drops showed appropriate reductions in bacterial viability after 6 hrs, 24 hrs and 7 days, but showed a very low bacterial recovery after 28 days which didn''t comply with the no recovery (NR) term of BP ''A'' criteria. Since viable microbial counts were usually determined by plate count method, it seems that the term of NR should define an acceptable range. The results indicated that there is not a clear correlation between antimicrobial efficacy testing of eye-drops and the rate of their microbial contamination while are being used. Other factors such as hygienic practices of eye-drops, proper bottle design and training of patients could influence their microbial contaminations. Regulation of in-use efficacy testing of eye-drops which is influenced by the environment, the frequency and technique of use, might be essential

Candidiasis associated with indwelling medical devices is especially problematic since they can act as substrates for biofilm growth which are highly resistant to antifungal drugs. Farnesol is a quorum-sensing molecule that inhibits filamentation and biofilm formation in Candida albicans. Since in recent years Candida tropicalis have been reported as an important and common non-albicans Candida species with high drug resistance pattern, the inhibitory effect of farnesol on biofilm formation by Candida tropicalis was evaluated. Five Candida tropicalis strains were treated with different concentration of farnesol (0, 30 and 300 µM) after 0, 1 and 4 hrs of adherence and then they were maintained under biofilm formation condition in polystyrene, 96-well microtiter plates at 37°C for 48 hrs. Biofilm formation was measured by a semiquantitative colorimetric technique based on reduction assay of 2,3- bis -2H-tetrazolium- 5- carboxanilide (XTT). The results indicated that the initial adherence time had no effect on biofilm formation and low concentration of farnesol (30 µM) could not inhibit biofilm formation. However the presence of non-adherent cells increased biofilm formation significantly and the high concentration of farnesol (300 µM) could inhibit biofilm formation. Results of this study showed that the high concentration of farnesol could inhibit biofilm formation and may be used as an adjuvant in prevention and in therapeutic strategies with antifungal drugs.

Background and purpose of the study: The acute respiratory distress syndrome (ARDS) is a common clinical disorder caused by injury to the alveolar epithelial and endothelial barriers of lung. In ARDS patients, oxidative stress is increased and plasma antioxidant levels are reduced. Vitamin E has an important role in antioxidant defense mechanisms. In this study the effect of vitamin E on decrease of APACHE II score in ARDS patients was investigated. Twenty patients [mean (SE): age = 51.2 ± 6.41 years] with ARDS were enrolled. After diagnosis based on inclusion and exclusion criteria, ten patients as treatment group received 600 IU vitamin E daily intramuscularly. Control group received normal saline as placebo. Plasma samples and Acute Physiology and Chronic Health Evaluation (APACHE) II score were obtained before administration, 4hrs and 12hrs after each intervention and repeated three days for each patient. Results were analyzed by use of an SPSS software package with a repeated-measures analysis of variance (ANOVA). Significant changes were observed in APACHE II score from first to seventh measurement (p=0.0001) in treatment group, but vitamin E concentration altered significantly in only first to seventh measurement (p= 0.019). From the results of this study, it seems that the use of vitamin E as a lipid-soluble antioxidant along with other supportive measures is beneficial in decreasing APACHE II score in ARDS patients.

Halothane and isoflurane inhibit glucose-induced insulin secretion in animal and in vitro experiments.A randomized trial was designed to determine their in vivo effects in diabetic patients. Sixty diabetic patients with various malignancies were randomized to receive anesthesia with either halothane or isoflurane. Blood glucose level (BGL) was measured before induction and every 30 minutes during surgery and then after operation in the recovery room. Patients in both arms matched with gender, age, operation time, and initial BGL. In both arms an increase in blood glucose level could be detected 30 minutes after induction of anesthesia, but higher in the halothane arm. This difference was maintained for the next 30 minutes as well as the rest of duration of the anesthesia. Both arms had higher blood glucose levels after operation compared to initial, pre-anesthesia state, and this increase was more conspicuous in the halothane arm. No Halothane toxicity was detected during hospitalization. Although the effects of stress hormones was not evaluated, halothane anesthesia caused a greater degree of hyperglycemia compared to isoflurane anesthesia. This might be secondary to halothane''s greater inhibitory effect.

Ornithogalum procerum Stapf. (Family: Liliaceae) is an Iranian medicinal plant found mainly in the east Azarbaiijan province of Iran. As part of our on-going phytochemical and bioactivity studies on medicinal plants from Iranian flora, various extracts and essential oils of this plant were analysed by GC-MS. The hydrodistillation of the ground aerial parts of O. procerum, Soxhlet-extraction of the dried and ground bulbs, and the hydrolysis of the methanol extract of the bulbs yielded, the essential oils, n-hexane extrcat and the hydrolysed methanolic extract respectively, which were analysed by the GC-MS.A total of 23 compounds were identified from the essential oils of the the aerial parts. The identified compounds represented 70.27% of the total essential oils. The main components of the aerial parts were phenylacetaldehyde (7.57%), hexahydrofarnesyl acetone (8.13%), docosan (5.52%) and 5-methyl octadecane (4.63%). From the n-hexan extract of the bulbs, seven hydrocarbons representing 99.39% of the total extract, were identified. Finally, from the hydrolyzed methanolic extract of the bulbs, four polysterol-type compounds accounting for 59.81% of the extract, were detected. The GC-MS analyses reavealed that the essential oils are mainly composed of oxygenated hydrocarbons, the n-hexane extract contains predominatly hydrocarbons, and the hydrolyzed methanolic extract comprises polysterol-type compounds.

Background and the purpose of study: Achillea talagonica Boiss. (Asteraceae) grows in the western and central parts of Iran. This plant has long been used in traditional medicine as an anti-inflammatory agent for treatment of rheumatic pain. Previously, the immunosuppressive activity of the aqueous extract of this endemic plant in experimental animals was reported. In this research, isolation of the main immunologically active components of A. talagonica, which were effective on humoral immune responses in BALB/c mice is elucidated. In order to find the main immunosuppressive components of A. talagonica, methanol and methanol-water (80% and 50% v:v) extracts were injected to BALB/c mice and the hemagglutinating antibody titer was assayed after immunization with SRBC (sheep red blood cells). Guided by this assay, active principles were separated by chromatographic methods. Isolated compounds were identified as caffeic acid 9-O-glucoside (1), quercetin (2), luteolin (3), 3''-methoxy luteolin (4), proline (5) and choline (6) by comparison of their spectral data with those of reported in literatures. Immunosuppressive property of choline (5 mgkg-1) was comparable to those of prednisolone (10 mgkg-1); although, quercetin (20 mgkg-1) and caffeoyl glucoside (20 mgkg-1) decreased anti-SRBC titer in comparison with control groups.Major Immunosuppressive effects of A. talagonica are due to some components belonging to betaine, flavonol and phenoilc esters.

In Iranian and other traditional medicines, an antidepressant effect has been indicated for Melissa officinalis (Lamiaceae). However, studies showing its antidepressant effect is lacking. Therefore, the present study was undertaken to examine whether the aqueous extract and essential oil from leaves of Melissa officinalis have an antidepressant-like activity in mice. The effect of subchronic administration of different doses of the aqueous extract (25, 75, 150, 300 mg/kg or water; n=9-10) and the essential oil (10, 25, 75, 150, 300 mg/kg or almond oil; n=9-10) on immobility, climbing, and swimming behaviors were evaluated in the forced swimming test. Fluoxetine (20mg/kg) and imipramine (15 mg/kg) were used as reference drugs. Additionally, the effect of both plant preparations on spontaneous activity was examined. All doses of the aqueous extract, used in this study, produced a significant reduction in immobility along with an increase in climbing behavior which is similar to those which have been observed with imipramine. Essential oil caused a dose-dependent reduction in immobility and an increase in climbing at all studied doses, compared to control group. Only the highest dose (300mg/kg) of essential oil showed a significant increase in swimming behavior. The aqueous extract, but not the essential oil, decreased spontaneous activity in a dose dependent manner. The results of this study suggests that the Melissa officinalis possess an antidepressant-like activity similar to imipramine which may have a potential clinical value for treatment of depression.

Nitric oxide (NO) have a dual action in mouse oocyte meiotic maturation which depends on its concentration, but the mechanisms by which it influences oocyte maturation has not been exactly clarified. In this study different signaling mechanisms which exist for in vitro maturation of meiosis was examined in cumulus cell-enclosed oocytes (CEOs) after injection of pregnant mare''s serum gonadotropin (PMSG) to immature female mice. The CEOs were cultured in spontaneous maturation and hypoxanthine (HX) arrested model. Sodium nitroprusside (SNP, an NO donor, 10mM) delayed germinal vesicle breakdown (GVBD) significantly during the first 5 hrs of incubation and inhibited the formation of first polar body (PB1) at the end of 24 hrs of incubation. SNP (10-5M) stimulated the meiotic maturation of oocytes significantly by overcoming the inhibition of HX. Sildenafil (a cGMP stimulator, 100 nM), had a significant inhibitory effects on both spontaneous meiotic maturation and HX-arrested meiotic maturation. Forskolin (an adenylate cyclase stimulator, 6µM) and SNP (10mM) had the same effects on GVBD. Forskolin reversed the SNP (10-5M) stimulated meiotic maturation. These results suggest that differences in pathways are present between SNP-inhibited spontaneous meiotic maturation and SNP-stimulated meiotic maturation in mouse oocytes

Background and the purpose of the study: Recent studies have indicated the role of apoptosis and angiotensin in the pathogenesis of bleomycin induced-pulmonary fibrosis. Losartan, an angiotensin type 1 receptor (AT1R) antagonist, has ameliorated apoptosis and fibrosis from bleomycin. In this study, alterations in the expression of apoptosis-regulatory genes (bcl-2 and bax) were investigated in different cells of lung tissue of mice treated with bleomycin in the presence of losartan. Losartan (10 mg/kg, i.p.) was given to mice two days before administration of bleomycin (3 U/kg) and throughout the test period. After two weeks, lung tissues of mice were evaluated for fibrosis by biochemical measurement of collagen deposition and semiquantitative analysis of pathological changes of the lung. The expression of bcl-2 and bax was assessed by immunohistochemical assay using biotin-streptavidin staining method on paraffin-embedded lung tissues.Results and major Pre-treatment with losartan significantly (P < 0.05) reduced the increase in lung collagen content and also inhibited the histological changes induced by bleomycin. Immunohistochemical studies showed that losartan significantly (P < 0.05) reduced the bax/bcl-2 expression ratio in the alveolar epithelial cells, lymphocytes, macrophages and interstitial myofibroblasts. Losartan also inhibited the bcl-2 upregulation which was educed by bleomycin in neutrophils. By reduction of bax/bcl-2 ratio as a determinant of susceptibility of a cell to apoptosis, losartan exerted protective effects on the alveolar epithelial cells that may be important in the amelioration of pulmonary fibrosis. These results may help to better understanding of the role of angiotensin II and apoptosis in pulmonary fibrosis.

Background and the purpose of the study: One of the major concerns about recombinant protein production is its possible toxicity for the organism. Purification of the recombinant protein is another challenge in this respect. Recently In Vitro translation cell free system that provides a coupled transcription-translation reaction for protein synthesis to overcome the above mentioned problems has been emerged. The aim of this study was expression of GFP as a marker for gene expression and protein in In Vitro translation system. pIVEX2.3-GFP plasmid was cloned to E. coli and the plasmid DNA extracted. In Vitro translation was performed with RTS 100 E. coli Hy kit according to manufacture''s instructions. Expression of recombinant fusion protein, His- GFP, was determined by SDS-PAGE, ELISA and western blot analysis. Expected size of recombinant protein was detected in SDS-PAGE and further confirmed by western blot analysis and ELISA.Major Results showed that In Vitro translation is suitable for expression of recombinant protein and fusion of the recombinant protein with His-tag facilitates the purification.