DARU, Journal of Pharmaceutical Sciences
Volume:17 Issue: 2, summer 2009

Background and the purpose of the study: The Reactive oxygen species (ROS) is induced in the cells following various stresses but the effect of recombinant protein expression on ROS generation has not been studied yet. In this study, H2O2 concentration and catalase activity variations and their correlation with cell growth following cytoplasmic expression of human interleukin-2 (hIL-2) and mouse interleukin-4 (mIL-4) in Escherichia coli were investigated. Additionally, the effect of recombinant protein expression under different conditions was compared to the effect of foreign DNA introduction on these factors. Plasmids pEThIL-2 and pETmIL-4 were used for expression of human interleukin-2 (hIL-2) and mouse interleukin-4 (mIL-4) inside the cytoplasm of the cells. Having confirmed protein expression using SDS-PAGE analysis and ELISA assay, H2O2 concentration and catalase activity were measured at various ODs.Results and major Empty vector introduction increased significantly H2O2 concentration of the cells. However, H2O2 concentration in hIL-2 and mIL-4 expressing cells was significantly higher than its concentration in empty vector transformed cells. Catalase activity was reduced in foreign DNA introduced cells. It was more lowered following expression of recombinant proteins. Results of this study revealed the relationship between foreign DNA introduction and protein expression with H2O2 elevation and catalase activity reduction. There was also correlation between H2O2 elevation and reduction in catalase activity with the cell growth depression.

Mast cells are believed to play a role in irritable bowel syndrome pathogenesis and symptom genesis due to their close neighborhood to gastrointestinal innervations. This study was designed to evaluate the efficacy of orally administered cromolyn for reduction of symptoms in patients with irritable bowel syndrome (IBS).Material and Methods s: A randomized placebo-controlled double-blinded 6×6 weeks cross-over study was performed in a private gastrointestinal clinic. 10 patients were allocated to group A and 6 patients to group B. Patients in group A received 150 mg cromolyn divided in three equal doses for the first 6 weeks and placebo for the next 6 weeks but patients in group B received placebo for the first 6 weeks and cromolyn in the next 6 weeks. Weekly evaluation was performed and visual analogue scale was used to determine severity of symptoms. Sixteen patients completed the study. Mean age of the patients was 40.3 ± 10.9 years old [range: 24-57]. Eight patients had D-IBS (Diarrhea dominant) and other 8 had C-IBS (Constipation dominant). Both cromolyn sodium and the placebo decreased the severity of bloating (Freidman test, p 0.001 and 0.006 respectively). The severity of the main symptom (diarrhea or constipation) did not decrease in patients of group A and B who were treated with different sequences of the drug or placebo. The severity of pain decreased drastically after 6th week of treatment with cromolyn. Freidman test showed a significant difference between the pain levels of the former defined treatment spots (p 0.01, and 0.02 for patients in group A and B, respectively). No adverse drug reactions were observed during the study. In conclusion, long term administration of cromolyn seems to be partially effective for treatment of abdominal pain in patients with IBS while main symptoms (diarrhea or constipation) might not decrease during this treatment.

Background and the purpose of the study: Midazolam is preferably used in pediatric dentistry for quick onset of action and recovery. The aim of this prospective, observer-blind and placebo-controlled study was to assess the efficacy of a low dose of oral midazolam in modification of the behavior of young pediatric dental patients. Forty children aged 3 to 5 years who displayed ratings 1 or 2 on the Frankl Scale and were healthy by the American Society of Anesthesiologists-I status were randomly divided into two experimental and control groups of 20 each. All children required pulpotomy and restoration of D and E teeth and received either 0.25mg/kg of a 15mg/3ml IV midazolam mixed in black cherry syrup or the syrup alone. Subjects were continuously observed and monitored with pulse oximetry. Houpt''s Behavioral Ratings was used to determine the overall behavior, the degree of crying and movement during treatment. Mann-Whitney U test was used for statistical analysis.Results and major Patients who received 0.25mg/kg of the prepared oral midazolam significantly behaved better during treatment than the placebo controls (P<0.05). In comparison with the placebo group, reduced movement and crying were observed in the midazolam group (P<0.05). No adverse effects were observed and treatments were completed successfully. A low dose of 0.25mg/kg of a 15mg/3ml IV midazolam mixed in black cherry syrup was found to be effective in conscious sedation of young pediatric dental patients.

Technetium-99m phytate (99mTc-ph) is a readily available radiopharmaceutical and has been suggested as a suitable agent for sentinel lymph node (SLN) detection. In this study, the diagnostic accuracy and false-negative rate of radionuclide SLN mapping using 99mTc-ph were investigated. Forty three women (mean age 52.3 years, range 31-74 years), who all had been diagnosed with breast cancer were enrolled in the study. All patients had no palpable axillary lymph nodes and had not undergone exploratory tumor resection or any drug treatment, previously. 99mTc-ph was injected peri-tumorally at four sites. Following SLN scintigraphic imaging, the patients were operated. Intraoperatively SLN were detected by a scintillation probe and a blue dye technique. Modified radical mastectomies with radical axillary dissection were performed with excision of the lymph nodes, to evaluate the accuracy of the SLN technique. Intraoperative scinti-mapping identified SLN in 40 of the 43 patients (detection rate: 93%). Scintigraphically, none of the patients had internal mammary drainage or contralateral axillary involvement. The blue dye detection rate in 23 patients under study was 87% and all lymph nodes detected by the blue dye technique were also detected as "hot" spots in the lymphoscintigraphy. Using pathology as the gold standard, the sensitivity and negative predictive value of scintigraphic lymphatic mapping in detection of SLN by 99mTc-ph were 90% and 90.9%, respectively. The same values for blue dye lymphatic mapping were 84.6% and 77.7%, respectively. 99mTc-ph used for SLN mapping is readily available, has low cost and gives better results than the blue dye technique. Long-term follow-up is required to assess accurately the incidence of failure in patients with negative SLN and the overall diagnostic accuracy and efficacy of the SLN mapping using 99mTc-ph as the radioactive tracer.

Influenza is a viral infectious disease with frequent seasonal epidemics causing world-wide economical and social effects. Due to antigenic shifts and drifts of influenza virus, long-lasting vaccine has not been developed so far. The current annual vaccines and effective antiviral drugs are not available sufficiently. Therefore in order to prevent spread of infectious agents including viruses, antiseptics are considered by world health authorities. Small particles of silver have a long history as general antiseptic and disinfectant. Silver does not induce resistance in microorganisms and this ability in Nano-size is stronger. The aim of this study was to determine antiviral effects of Nanosilver against influenza virus. TCID50 (50% Tissue Culture Infectious Dose) of the virus as well as CC50 (50% Cytotoxic Concentration) of Nanosilver was obtained by MTT (3- [4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl-tetrazolium bromide, Sigma) method. This compound was non-toxic to MDCK (Madin-Darbey Canin Kidney) cells at concentration up to 1 µg/ml. Effective minimal cytotoxic concentration and 100 TCID50 of the virus were added to the confluent cells. Inhibitory effects of Nanosilver on the virus and its cytotoxicity were assessed at different temperatures using Hemagglutination (HA) assay, RT-PCR (Reverse Transcriptase-Polymerase Chain Reaction), and DIF (Direct Immunofluorescent). RT-PCR and free band densitometry software were used to compare the volume of the PCR product bands on the gel. In this study it was found that Nanosilver has destructive effect on the virus membrane glycoprotein knobs as well as the cells.

During the last decade, the incidence of fungal infection has been increased in many countries. Because of the advent of resistant to antifungal agents, determination of an efficient strategic plan for treatment of fungal disease is an important issue in clinical mycology. Many methods have been introduced and developed for determination of invitro susceptibility tests. During the recent years, flow cytometry has developed to solving the problem and many papers have documented the usefulness of this technique. As the first step, the invitro susceptibility of standard PTCC (Persian Type of Culture Collection) strain and some clinical isolates of Candida consisting of Candida albicans, C. dubliniensis, C. glabrata, C. kefyer and C. parapsilosis were evaluated by macrodilution broth method according to NCCLS (National Committee for Clinical Laboratory Standards) guidelines and flow cytometry susceptibility test. The data indicated that macro dilution broth methods and flow cytometry have the same results in determination of MIC (Minimum Inhibitory Concentration) for amphotericin B, clotrimazole, fluconazole, ketoconazole and miconazole in C. albicans PTCC 5027 as well as clinical Candida isolates, such as C.albicans, C.dubliniensis, C.glabrata C.kefyr, and C.parapsilosis. Comparing the results obtained by macrodilution broth and flow cytometry methods revealed that flow cytometry was faster. It is suggested that flow cytometry susceptibility test can be used as a powerful tool for determination of MIC and administration of the best antifungal drug in treatment of patients with Candida infections.

Umbelliprenin, the natural prenylated coumarin distributed in the plants of apiaceae family, has shown various biological activities, especially as a cancer chemopreventive agent. In the present study, umbelliprenin, was examined for in vitro antioxidant activity, in vitro inhibitory activity against lipoxygenase, and in vivo anti-inflammatory activity. The applied tests were interaction with 1,1-diphenyl-2-picryl-hydrazyl (DPPH) stable free radical, inhibition of lipid peroxidation, inhibition of soybean lipoxygenase and in vivo inhibition of the carrageenin-induced rat paw edema. Umbelliprenin did not show any significant antioxidant activity but exhibited a remarkable and potent inhibition against soybean lipoxygenase (IC50 = 0.0725 μM). This compound, in the in vivo anti-inflammatory test, could also inhibit the carrageenin induced paw edema significantly (39 %). The observed inhibition of lipoxygenase may be a plausible mechanism for the potent cancer chemopreventive activity of umbelliprenin and may pose this compound as a valuable agent for the treatment of inflammatory diseases.

Background and the purpose of the study: Date palms (Phoenix dactylifera L., Arecaceae) are one of the oldest cultivated plants which are used in folk medicine for treatment of various diseases. Due to the presence of antioxidant compounds in this plant and the role of oxidative stress in pathogenesis of chronic diseases, the aim of this study was to determine the antioxidant activity and total phenolic contents of date palm fruits extracts with water, methanol 50%, DMSO, and mixture of water-methanol-acetone-formic acid (20:40:40:0.1). Antioxidant activity of extracts were measured by two tests: inhibition of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and FRAP (Ferric Reducing Antioxidant Power).Results and major Among 10 different varieties which were examined, the DMSO extract of Khenizi showed the highest antioxidant activity with the FRAP value of 3279.48 µmol/100 g of the dry plant and DPPH inhibitory percentage of 56.61%. DPPH scavenging radical and FRAP values of some varieties including Khenrizi, Sayer, Shahabi and Maktub showed a significant increase and were comparable to α-tocopherol (10 mg/L) when extracted by DMSO. Formic acid extract of Shahabi variety with 276.85 mg GAE/100 g of the dry plant showed the highest total phenolic content compared to other varieties. There was no correlation between accumulation of total phenol and antioxidant activity of extracts, explaining existence of other antioxidant components in date.

Background and the purpose of the study: Ghrelin increases food intakes and body weight via growth hormone secretagogues receptor (GHSR-Ia). [D-Arg-1, D-phe-5, D-Trp-7,9, Leu-11]Substance-P (an analog of Substance-P) is known as an antagonist of GHSR-Ia receptor which inhibits ghrelin- induced food intakes. Thyroid hormones have also an important role in the regulation of metabolism and body weight. The goal of this study was to determine the effect of different doses of either ghrelin, analog of Substance-P or their interactions on the body weight, mean plasma TSH, Triiodothyronine (T3) and Thyroxine (T4) concentration and to investigate whether ghrelin exerts its effects on thyroid axis activity via GHSR-Ia. Rats received different doses of ghrelin (1, 5, or 10 nmol), analog of Substance P (5, 10, 20 nmol) or saline via lateral cerebral ventricle. Body weight was measured daily before injection and at 9 h of the day after the final injection. The blood samples were collected at the end of experiment. Plasma was assayed for thyroid hormones concentration. Ghrelin significantly increased body weight and decreased mean plasma TSH, T3 and T4 concentrations in a dose related manner. Analog of Substance-P significantly decreased body weight and increased thyroid hormones concentration in a dose related manner. It blocked the stimulatory effect of ghrelin on body weight and the inhibitory effect of ghrelin on thyroid axis activity. From the results of this study it appears that analog of substance P may be useful for treatment of obesity.

The effects of chronic lithium exposure on spatial memory in rats remain controversial. In this study a time course of the effects of lithium, administered systemically, on spatial memory acquisition in Morris water maze was investigated. Lithium (600 mg/L) was administered to four groups of rats in their drinking water; the first group of animals received lithium for one week, the second group for two weeks, the third group for three weeks, and the fourth group for four weeks. As controls, four groups of animals received only normal drinking water for the same period of time. Toward the end of their lithium or water treatment, all animals were trained for four days; each day included one block and each block contained four trials. Test trials were conducted 48 hrs after completion of the lithium treatment. Escape latency, traveled distance and swimming speed were evaluated during testing trials. Brain tissues from animals were processed according to the standard protocols for immunohistochemical analysis. Lithium treatment decreased escape latency and traveled distance, but not swimming speed, compared with controls, suggesting significant spatial memory acquisition enhancement by lithium. Quantitative analysis showed that lithium, particularly after four weeks of exposure, significantly increased the number and density of immunostained ChAT-containing (choline acetyltransferase) neurons in the medial septal area in comparison with control groups. There was also a significant correlation between the number of immunostained ChAT neurons and behavioral measures. These results suggest that chronic oral administration of lithium causes spatial memory acquisition improvement in rats and an increase in ChAT immunostaining levels in medial septal nuclei.

Large inter-individual variability has been reported for vancomycin pharmacokinetics in pediatric patients. On the other hand, the pharmacokinetic parameters of vancomycin should be known in order to individualize its dosage regimen. Therefore, this study was designed and conducted to assess the steady-state vancomycin serum concentration and pharmacokinetics in a population of Iranian pediatric patients. Vancomycin serum concentration at steady-state was determined in 62 children who were treated with vancomycin intermittent intravenous infusion. Also individual steady-steady pharmacokinetic parameters (total body clearance, apparent volume of distribution and elimination half-life) were determined in 30 patients who had both peak and trough vancomycin levels assuming one-compartment model. Calculated pharmacokinetic parameters were compared among patients with different underlying diseases and also with the results of similar studies that used one-compartment pharmacokinetic model for description of serum concentration of vancomycin at steady-state. More than half of the measured vancomycin serum concentrations were outside the recommended therapeutic range. Median trough concentration was significantly lower in critically ill patients as compared to patients of other disease categories. Although critically care patients showed greater values of apparent volume of distribution and also vancomycin clearance, no statistically significant difference of the calculated pharmacokinetic parameters could be detected among different groups of patients. While calculated volume of distribution for patients of this study was greater than those of similar studies, this difference could not be considered statistically significant in the majority of disease categories It may be concluded that design of vancomycin dosage regimens according to the recommended and general guidelines in literature (e.g. based on patient creatinine clearance) could not result in the desired therapeutic serum concentrations in the study population.

Background and purpose of the study: Several methods are available for control release of propranolol hydrochloride (PLH). The aim of the present study was to develop a novel technique to sustain PLH release from matrices. Matrices of PLH containing sodium carboxymethylcellulose (Na CMC) and various amounts of the inorganic cations Ca2+ and Al3+ were prepared. Dissolution of the matrices was carried out using the USP apparatus I. Analysis of release data was performed by some model independent and dependent approaches. The release of PLH was affected by incorporation of different amounts (milliequivalents, meq) of Ca2+ and Al3+. When the Ca2+amount increased from 0- 0.375 meq, the fraction of PLH which released within 480 min was augmented from 0.74 to 1 apparently via disintegrating effect of the cation. Al3+in the range 0- 0.125 meq, decreased the fractional release from 0.74 to 0.37 presumably by in situ cross- linking with polymer. Al3+ between 0.125 and 0.5 meq enhanced the release from 0.37 to 1 possibly due to the disintegrating effect. Among model independent metrics, the mean release time (MRT) failed to represent the effect of the cations on the release but the release efficiency (RE) as well as a suggested mean release rate (MRR) correlated well with the experimental release rate. Due to the complexity of the release, the only suitable kinetic model was the Weibull distribution. The minimum and maximum Weibull release rate constants for matrices containing Al3+ were 0.0007-0.017 1/min. The corresponding values for the matrices with Ca2+ were 0.0029-0.0082 1/min. Through careful choice of the amount of Al3+in NaCMC matrices the release of PLH can be controlled at a desired rate. The best model independent approach is MRR and the most accurate model dependent method is Weibull distribution to describe the release data.