Advanced Pharmaceutical Bulletin
Volume:3 Issue: 1, Jun 2013

Since its introduction capillary electrophoresis has shown great potential in areas where electrophoretic techniques have rarely been used before, including here the analysis of pharmaceutical substances. The large majority of pharmaceutical substances are neutral from electrophoretic point of view, consequently separations by the classic capillary zone electrophoresis; where separation is based on the differences between the own electrophoretic mobilities of the analytes; are hard to achieve. Micellar electrokinetic capillary chromatography, a hybrid method that combines chromatographic and electrophoretic separation principles, extends the applicability of capillary electrophoretic methods to neutral analytes. In micellar electrokinetic capillary chromatography, surfactants are added to the buffer solution in concentration above their critical micellar concentrations, consequently micelles are formed; micelles that undergo electrophoretic migration like any other charged particle. The separation is based on the differential partitioning of an analyte between the two-phase system: the mobile aqueous phase and micellar pseudostationary phase. The present paper aims to summarize the basic aspects regarding separation principles and practical applications of micellar electrokinetic capillary chromatography, with particular attention to those relevant in pharmaceutical analysis.

Behçet Disease (BD) is an autoimmune disorder with recurrent ocular, vascular, central nervous system, articular, mucocutaneous, and gastrointestinal manifestations with unclear etiology and pathogenesis. The further characterization of inflammatory features of Behçet’s disease may eventually lead to development of better treatment options. Clinical and laboratory observations suggested an important role of IL-17, IL-21 and neutrophil-mediated process in the pathogenesis of BD. New therapeutic modalities target specific and nonspecific suppression of the immune system. The various non-specific immunosuppressive drugs, used either alone or in combinations, frequently fail to control inflammation or maintain remissions. Due to encouraging clinical results (i.e. Antigenic specification, prolonged survival with acceptable levels of toxicity); antibody-based drugs could be effective for the clinical management of Behçet’s disease.

In the present study in vivo analgesic activity of some previously synthesized 1,2,4-triazole derivatives containing pyrazole, tetrazole, isoxazole and pyrimidine ring have been evaluated. Acetic acid induced writhing method and Hot plate method has been described to study analgesic activity of some 1,2,4-triazole derivatives containing pyrazole, tetrazole, isoxazole and pyrimidine as a pharmacological active lead. Thirty six different derivatives containing 1,2,4-triazole ring were subjected to study their in vivo analgesic activity. Chloro, nitro and methoxy, hydroxy and bromo substituted derivatives showed excellent analgesic activity and dimethylamino, furan and phenyl substituted derivatives showed moderate analgesic activity in both of the methods. Compounds IIIa, IIId, IIIf, IIIi, IIIj, IVa, IVb, IVd, IVf, IVh, IVj IV3a and IIj were found to be superior analgesic agents after screening by Acetic acid induced writhing method. Compounds IIIb, IIId, IIIf, IIIh, IIIj, IVa, IVb, IVd, IVf, IVh, IVi, IV3c, IV3e and IIj were showed analgesic potential after screening of Hot plate method. All tested compounds containing 1,2,4-triazole were found to be promising analgesic agents, for this activity pyrazole, tetrazole, isoxazole and pyrimidine leads might be supported.

Recombinant tumor necrosis factor-alpha (TNF-α) has been utilized as an antineoplastic agent for the treatment of patients with melanoma and sarcoma. It targets tumor cell antigens by impressing tumor-associated vessels. Protein purification with affinity chromatography has been widely used in the downstream processing of pharmaceutical-grade proteins. In this study, we examined the potential of our produced anti-TNF- scFv fragments for purification of TNF-α produced by Raji cells. he Raji cells were induced by lipopolysaccharides (LPS) to express TNF-α. Western blotting and Fluorescence-activated cell sorting (FACS) flow cytometry analyses were used to evaluate the TNF-α expression. The anti-TNF-α scFv selected from antibody phage display library was coupled to CNBr-activated sepharose 4B beads used for affinity purification of expressed TNF-α and the purity of the protein was assessed by SDS-PAGE. Western blot and FACS flow cytometry analyses showed the successful expression of TNF-α with Raji cells. SDS-PAGE analysis showed the performance of scFv for purification of TNF-α protein with purity over 95%. These findings confirm not only the potential of the produced scFv antibody fragments but also this highly pure recombinant TNF-α protein can be applied for various in vitro and in vivo applications.

The major limitation in the widespread use of liposome drug delivery system is its instability. Lyophilization is a promising approach to ensure the long-term stability of liposomes. The aim of this study was to prepare sirolimus-loaded liposomes, study their stability and investigate the effect of lyophilization either in the presence or in the absence of lyoprotectant on liposome properties. Two types of multi-lamellar liposomes, conventional and fusogenic, containing sirolimus were prepared by modified thin film hydration method with different ratio of dipalmitoylphosphatidylcholine (DPPC), cholesterol and dioleoylphosphoethanolamine (DOPE), and were lyophilized with or without dextrose as lyoprotectant. Chemical stability investigation was performed at 4°C and 25°C until 6 months using a validated HPLC method. Physical stability was studied with determination of particle size (PS) and encapsulation efficiency (EE %) of formulations through 6 months. Chemical stability test at 4°C and 25°C until 6 months showed that drug content of liposomes decreased 8.4% and 20.2% respectively. Initial mean EE % and PS were 72.8 % and 582 nm respectively. After 6 months mean EE % for suspended form, lyophilized without lyoprotectant and lyophilized with lyoprotectant were 54.8 %, 62.3% and 67.1 % at 4°C and 48.2%, 60.4 % and 66.8 % at 25°C respectively. Corresponding data for mean PS were 8229 nm, 2397 nm and 688nm at 4°C and 9362 nm, 1944 nm and 737 nm at 25°C respectively. It is concluded that lyophilization with and without dextrose could increase shelf life of liposome and dextrose has lyoprotectant effect that stabilized liposomes in the lyophilization process.

This study aimed to evaluate the possible estrogenic activity of some ingredients of Nigella sativa including Linoleic acid and Gama-Linolenic acid by vaginal cornification assay. Forty ovariectomized (OVX) rats, aged 16 weeks were allotted randomly to five groups: negative control (taking 1 ml olive oil/ day); positive control (taking 0.2 mg/kg/day Conjucated Equine Estrogen-CEE); experimental groups (taking 50 mg/kg/day Linoleic acid or 10 mg/kg/day Gamma Linolenic acid or 15mg/kg/day Thymoquinone). All of supplements administered via intragastric gavage for 21 consecutive days. To assess estrogen like activity, vaginal smear was examined daily and serum estradiol was measured at baseline, after 10 days and at the end of experiment. The significant occurrence of vaginal cornification cell (p<0.05) after Linoleic acid supplementation indicated estrogenic activity of Linoleic acid which was in consistency with serum estradiol level, but this effect was not as much as CEE. Gama-Linolenic acid also exist a few cornified cell in smear which was not significantly differ from those control group. Linoleic acid showed the beneficial effects on OVX rats’ reproductive performance, thereby indicating its beneficial role in the treatment of the postmenopausal symptoms.

Piroxicam, is non–steroidal anti–inflammatory and analgesic agent, which is widely used in the treatment of patients with rheumatologic disorders. A new analytical approach based on the dispersive liquid–liquid microextraction (DLLME) has been developed for the extraction and determination of PX in pharmaceutical preparation and human urine. From the PX standard solution or solutions prepared from real samples, aliquot volumes were pipetted into centrifuge tubes and mixed with acetate buffer at pH 3.0 and NaCl solution. The contents were subjected to the DLLME, so 700 μL of methanol containing 70 μL of chloroform was injected rapidly into a sample solution. A cloudy solution was rapidly produced and the PX extracted into dispersed fine droplets. The mixture was centrifuged, thus these fine droplets of chloroform were settled. The supernatant aqueous phase was readily decanted, then the remained organic phase was diluted with ethanol and the absorbance measured at 355 ± 3 nm against a reagent blank. The main factors affecting the extraction efficiency such as pH, extraction and disperser solvent types and etc. were studied and optimized systematically. Under optimized conditions, the calibration graphs were linear over the range of 0.2 to 4.8 μg/mL. The limit of detection and relative standard deviation were found to be 0.058 μg/mL and 2.83%, respectively. Relative recoveries in the spiked samples ranged from 97 to 110%. Using the developed method PX can be analyzed in pharmaceutical formulation and human urine sample in a simpler, cheaper and more rapid manner.

HIV/AIDS is a major public health issue. Studies have shown that young people between the ages of 15 to 24 years accounts for more than 40% of new HIV infections. However, new infections could be prevented if the desired impact is made on young people through interventions such as effective peer education program to improve their knowledge, attitude and practice. Therefore, this study is aimed at assessing the impact of peer education vis-a-vis attitudinal changes and practices on issues relating to HIV/AIDS among youths in North central Nigeria. This classical experimental study was carried out in three stages, that is: pre-intervention, intervention and post-intervention stages among students of Government High School, Ilorin between the ages of 15 and 24 years. The sample size was 80 students each for study and control groups. Proportional gender distribution was ensured in the selection and the sampling technique was multistage sampling technique. Data collection was through semi structured self administered pre and post evaluation questionnaire. Peer education training and forth night mentoring sessions was conducted for the study group. Impact of the intervention was assessed after eight weeks. Analysis of the result shows that out of the sixteen questions asked on respondents‟ attitude, only four showed statistical significance between the study and control group before the intervention which shot up to eleven after intervention. Also, the frequency of those having bad practices regarding prevention of HIV/AIDS also reduced after intervention. This reduction was much more in many instances when compared to the control group. This study has revealed that peer education training among youth can go a long way in educating them on issues relating to HIV/AIDS prevention. It can also bring about attitudinal changes and better practices that may help in reducing incidence and prevalence of HIV/AIDS in our society.

Salvia officinalis L., also known as Maryam Goli, is one of the native plants used to Persian medicinal herbs. Hence, the objective of this study was to examine the in vitro cytotoxic activities of a standardized crude methanol extracts prepared from Salvia officinalis L., on a non-Hodgkin’s B-cell lymphoma (Raji) and human leukemic monocyte lymphoma (U937), Human acute myelocytic leukemia (KG-1A) and Human Umbilical Vein Endothelial (HUVEC) cell lines. The effect of methanolic extract on the inhibition of cell proliferation and cytotoxic activity was evaluated by Dye exclusion and Micro culture tetrazolium test (MTT) cytotoxicity assay. Cell death ELISA was employed to quantify the nucleosome production result from nuclear DNA fragmentation during apoptosis and determined whether the mechanism involves induction of apoptosis or necrosis. The present results demonstrated that methanolic extract at 50 to 800 μg/ml dose and time-dependently suppressed the proliferation of KG-1A, U937 and Raji cells by more than 80% (p<0.01), with ascending order of IC50 values in 24: KG-1A (214.377 μg/ml), U937 (229.312 μg/ml) and Raji (239.692 μg/ml) when compared with a chemotherapeutic anticancer drug, paclitaxel (Toxol), confirming the tumour-selective cytotoxicity. The crude extract however did not exert any significant cytotoxic effect on normal cell line HUVEC (IC50>800 Ag/ml). Nucleosome productions in KG-1A, Raji and U937 cells were significantly increased respectively upon the treatment of Salvia officinalis L. extract. The Salvia officinalis L. extract was found dose and time-dependently inhibits the proliferation of lymphoma and leukemic cells possibly via an apoptosis-dependent pathway.

The purpose of this research was to analyze cagA and vacA genotypes status in H. pylori isolates and relationship with clinical outcomes. Gastric biopsy specimens were cultured for H. pylori isolation and cagA and vacA genes were detected in these isolates. Data were collected and the results were analyzed using χ2 and Fishers exact tests by SPSS software version. 16. Of the total 115 H. pylori isolates, 79 (68.7 %) were cagA positive and 82 (71.3%) of isolates contained the s1 allele which 33 (28.7%) were subtype s2. s1m2 was the most frequent vacA allelic combination in the H. pylori isolates examined (63 cases), followed by s2m2 (31 cases), s1m1 (19 cases) and s2m1 (2 case). Strains cagA positive were more frequent in peptic ulcer diseases patients than non ulcer diseases patients, as 47 (59.5%) and 32 (40.5%), while cagA negative were low, as 15 (41.7%) and 21 (58.3%), respectively. We found that the cagA and vacA status were not related to clinical outcomes in this area. Overall, in the present study, vacA s1/m2, cagA-positive strains were predominant irrespective of clinical outcome, but s2/m1 was rare.

This research was aimed to formulate and characterize a microemolsionsystems as a topical delivery system of naproxen for relief of symptoms of rheumatoidarthritis, osteoarthritis and treatment of dysmenorrheal. ME formulations prepared by mixing of appropriate amount of surfactant including Tween 80 and Span 80, co-surfactant such as propylene glycol (PG) and oil phase including Labrafac PG – transcutol P (10:1 ratio). The prepared microemolsions were evaluated regarding their particle size, zeta potential, conductivity, stability, viscosity, differential scanning calorimetry (DSC), scanning electron microscopy (SEM), refractory index (RI) and pH. The mean droplets size of microemulsion formulation were in the range of 7.03 to 79.8 nm, and its refractory index (RI) and pH were 1.45 and 6.75, respectively. Viscosity range was 253.73- 802.63cps. Drug release profile showed that 26.15% of the drug released in the first 24 hours of experiment. Also, Hexagonal and bicontinuous structures were seen in the SEM photograph of the microemulsions. characterization, physicochemical properties and in vitro release were dependent upon the contents of S/C ratio, water and, oil phase percentage in formulations. Also, ME-6 may be preferable for topical naproxen formulation.

Farnesyltransferase (FTase) is a zinc-dependent enzyme that adds a farnesyl group to the Ras proteins. L778, 123 is a potent peptidomimetic imidazole-containing FTase inhibitor. L778123 was synthesized according to known methods and evaluated alone and in combination with doxorubicin against A549 (adenocarcinomic human alveolar basal epithelial cells) and HT29 (human colonic adenocarcinoma) cell lines by MTT assay. L778123 showed weak cytotoxic activity with IC50 of 100 and 125 for A549 and HT-29 cell lines, respectively. The combination of doxorubicin and L778123 can decrease IC50 of doxorubicin in both cell lines significantly. It can be concluded that L778, 123 can be a good agent for combination therapy.

In this work a numerical method, based on the use of spectrophotometric data coupled to partial least squares (PLS) regression and net analyte preprocessing combined with classical least square (NAP/CLS) multivariate calibration, is reported for the simultaneous determination of metformin hydrochloride (MET), gliclazide (GLZ) and pioglitazone hydrochloride (PIO) in synthetic samples and combined commercial tablets. Spectra of MET, GLZ and PIO were recorded at concentrations within their linear ranges (5-25 μg/ml, 0.5-8 μg/ml and 0.5-3 μg/ml respectively) and were used to compute a total of 25 synthetic mixtures involving 15 calibration and 10 validation sets between wavelength range of 200 and 400 nm in 0.1N HCl. The suitability of the models was decided on the basis of root mean square error (RMSE) values of calibration and validation data. The analytical performances of these chemometric methods were characterized by relative prediction errors and recovery studies (%) and were compared with each other. These two methods were successfully applied to pharmaceutical formulation, tablet, with no interference with excipients as indicated by the recovery study results. Mean recoveries of the commercial formulation set together with the figures of merit (calibration sensitivity, selectivity, limit of detection, limit of quantification etc.) were estimated. The proposed methods are simple, rapid and can be easily used as an alternative analysis tool in the quality control of drugs and formulation.

The objective of this study was to prepare a suitable form of nanofiber for indomethacin using polymers Eudragit RS100 (ERS) and Eudragit S100 (ES) and to evaluate the effect of some variables on the characteristics of resulted electrospunnanofibers. Electrospinning process was used for preparation of nanofibers. Different solutions of combinations of ERS, ES and indomethacin in various solvents and different ratios were prepared. The spinning solutions were loaded in 10 mL syringes. The feeding rate was fixed by a syringe pump at 2.0 mL/h and a high voltage supply at range 10-18 kV was applied for electrospinning. Electrospunnanofibers were collected and evaluated by scanning electron microscopy, differential scanning calorimetry and FTIR for possible interaction between materials used in nanofibers. The effect of solvent and viscosity on the characteristics of nanofibers also was investigated. Fiber formation was successful using a solvent ethanol and mixture of ERS and ES. Increase in viscosity of ethanolic solutions of ERS followed by addition of ES in the solution led to preparation of smooth fibers with larger diameters and less amounts of beads. DSC analysis of fibers certified that indomethacin is evenly distributed in the nanofibers in an amorphous state. FTIR analysis did not indicate significant interaction between drug and polymer. It was shown that drug-loaded ERS and ES nanofibers could be prepared by exact selection of range of variables such as type of solvent, drug: polymer ratio and solution viscosity and the optimized formulations could be useful for colonic drug delivery.

Diabetes is associated with memory and learning disorder. The purpose of this study is to determine the effect of acute oral administration of loganin on memory in diabetic male rats. 42 male Wistar rats (250-300 g) were divided into six groups: Control, Diabetic (1 week), Diabetic (12 weeks), Loganin, Diabetic (1 week) + Loganin, Diabetic (12 weeks) + Loganin. Diabetes was induced by IP injection of Streptozotocin (60 mg/kg). Loganin (40 mg/kg, po) was administrated 1 hour before test. Then, spatial memory was compared between groups with Morris Water Maze tests. Administration of loganin during acquisition, significantly (p<0.05) decreased both escape latency and traveled distance to find hidden platform in 1 and 12 weeks diabetic rats. In evaluation of recall phase of memory, loganin significantly (p<0.05) increased time and distance spent in the target quadrant in 1 and 12 weeks diabetic rats. Acute administration of loganin could improve spatial memory in diabetic rats.

The aim of this study was to determine the toxic effect of Nigella sativa powder on the liver function which was evaluated by measuring liver enzymes and through histopathological examination of liver tissue. Twenty four male Sprague Dawley rats were allotted randomly to four groups including: control (taking normal diet); low dose (supplemented with 0.01 g/kg/day Nigella sativa); normal dose (supplemented with 0.1 g/kg/day Nigella sativa) and high dose (supplemented with 1 g/kg/day Nigella sativa). All of supplements administered in powder form mixed with rats’ pellet for 28 days. To assess liver toxicity, liver enzymes measurement and histological study were done at the end of supplementation. The finding revealed that there was no significant change in serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) between treatment groups. Histopathological study showed very minimal and mild changes in fatty degeneration in normal and high doses of Nigella sativa treated group. Inflammation and necrosis were absent. The study showed that supplementation of Nigella sativa up to the dose of 1 g/kg supplemented for a period of 28 days resulted no changes in liver enzymes level and did not cause any toxicity effect on the liver function.

The present study was designed to investigate the anti-nociceptive activity of ethanolic leaf extract of Trema cannabina Lour (family: Cannabaceae) in experimental animal models.

The anti-nociceptive action was carried out against two types of noxious stimuli, thermal (hot plate and tail immersion tests) and chemical (acetic acid-induced writhing) in mice.

Phytochemical analysis of crude extract indicated the presence of reducing sugar, tannins, steroid and alkaloid types of secondary metabolites. Crude extract of T. cannabina (500 mg/kg dose) showed maximum time needed for the response against thermal stimuli (6.79±0.15 seconds) which is comparable to diclofenac sodium (8.26±0.14 seconds) in the hot plate test. Hot tail immersion test also showed similar results as in hot plate test. At the dose of 250 and 500 mg/kg body weight, the extract showed significantly and in a dose-dependent (p<0.001) reduction in acetic acid induced writhing in mice with a maximum effect of 47.56% reduction at 500 mg/kg dose comparable to that of diclofenac sodium (67.07%) at 25 mg/kg.

The obtained results tend to suggest the Anti-nociceptive activity of ethanolic leaf extract of Trema cannabina and thus provide the scientific basis for the traditional uses of this plant part as a remedy for pain.

Monoclonal antibodies are important tools are used in basic research as well as, in diagnosis, imaging and treatment of immunodeficiency diseases, infections and cancers. The purpose of this study was to produce large scale of monoclonal antibody against CD20 in order to diagnostic application in leukemia and lymphomas disorders. Hybridoma cells that produce monoclonal antibody against human CD20 were administered into the peritoneum of the Balb/c mice which have previously been primed with 0.5 ml Pristane. After twelve days, approximately 7 ml ascetic fluid was harvested from the peritoneum of each mouse. Evaluation of mAb titration was assessed by ELISA method. In the present study, we describe a protocol for large scale production of MAbs. We prepared monoclonal antibodies (mAbs) with high specificity and sensitivity against human CD20 by hybridoma method and characterized them by ELISA. The subclass of antibody was IgG2a and its light chain was kappa. Ascetic fluid was purified by Protein-A Sepharose affinity chromatography and the purified monoclonal antibody was conjugated with FITC and Immunofluorescence was done for confirming the specific binding. The conjugated monoclonal antibody could have application in diagnosis B-cell lymphomas, hairy cell leukemia, B-cell chronic lymphocytic leukemia, and melanoma cancer stem cells.

The aim of present in vitro studies was performed to examine the antioxidant and anticancer activities of ethanol and aqueous extracts of Drosera indica L. Different concentrations (5 – 640mcg/ml) of the ethanol (EEDI) and aqueous (AEDI) extracts of D.indica L were used in various antioxidant assay methods such as hydroxyl radicals, DPPH, super oxide radical scavenging activity, chelating ability of ferrous ion, nitric oxide radical inhibition, ABTS and reducing power. Ascorbic acid (AA) was used as the standard antioxidant for the free radical scavenging assays. Dalton’s Ascitic Lymphoma (DAL) and Ehrlich Ascitic Carcinoma (EAC) cell lines were used as the in vitro cancer models for the tryphan blue dye and LDH leakage assays, where 5 to 250mcg /ml of both EEDI and AEDI were tested. EEDI showed antioxidant activities with the minimum IC50 values of 34.8±0.43 mcg/ml in scavenging of hydroxyl radical and moreover AEDI showed minimum IC50 values of 94.51±0.84 mcg/ml in Fe2+chelating assay. EEDI on the reducing power assay and ABTS showed higher IC50 than standard AA. IC50 values of AEDI on Fe2+ chelating assay and super oxide radical assay was lesser than IC50 value of AA. Both extracts at 250mcg/ml dose showed remarkable increase in the percentage of dead cancer cells (90% by EEDI and 86% by AEDI in DAL model and 89% by EEDI and 80% by AEDI in EAC model). It is concluded from this study that D.indica L exhibited excellent antioxidant activity against the different in vitro antioxidant models and anticancer activity against the two different cell lines tested.

The antimicrobial activity of doripenem in comparison of imipenem, meropenem and ertapenem among Pseudomonas aeruginosa isolated from burn and Cystic Fibrosis (CF) patients were determined. Metallo-β-lactamase (MBL) genes in imipenem non susceptible P. aeruginosa isolates were detected using PCR method. The in vitro susceptibilities of doripenem, imipenem, meropenem and ertapenem were determined by Etests. MIC50 and MIC90 for corresponding antibiotics were determined individually in burn and CF isolates. Among isolates which were resistant to imipenem, 16 isolates were positive for the bla IMP gene. All isolates had no bla VIM gene. All MBL producing isolates were excluded. MIC50/MIC90 of doripenem in CF and burn isolates were 0.75/>32 and >32/>32 mg/L respectively. The corresponding values for imipenem in CF and burn isolates were 2/>32 and >32/>32 mg/L, respectively. The susceptibility rate of doripenem is higher than that of imipenem and meropenem among P.aeruginosa isolated from CF patients, whereas, there is no difference between the efficiency of doripenem and old carbapenems in non MBL producing P.aeruginosa isolates in burn patients.

Telomerase is expressed in most cancers, including breast cancer. Curcumin, a polyphenolic compound that obtained from the herb of Curcuma longa, has many anticancer effects. But, its effect is low due to poor water solubility. In order to improve its solubility and drug delivery, we have utilized a β-cyclodextrin-curcumin inclusion complex. To evaluate cytotoxic effects of cyclodextrin-curcumin and free curcumin, MTT assay was done. Cells were treated with equal concentration of cyclodextrin-curcumin and free curcumin. Telomerase gene expression level in two groups was compared by Real-time PCR. MTT assay demonstrated that β-cyclodextrin-curcumin enhanced curcumin delivery in T47D breast cancer cells. The level of telomerase gene expression in cells treated with cyclodextrin-curcumin was lower than that of cells treated with free curcumin (P=0.001). Results are suggesting that cyclodextrin-curcumin complex can be more effective than free curcumin in inhibition of telomerase expression.

The present study was designed to evaluate the occurrence of Legionella contamination in the tap water of Tabriz hospitals, Azerbaijan, Iran. One hundred and forty water samples from diverse water supply systems of 17 hospitals were collected and analyzed for the presence of Legionella spp. by PCR assay. In this study, 10 of 140 (7.1%) samples were positive for Legionella which L. pneumophila was detected in 4 (2.85%) water samples. In conclusion, hospital potable systems are the primary reservoirs for Legionnaires’ disease. This study concludes that Legionella spp. are present in aquatic hospitals environment of Tabriz. Due to the serious risk of infections, it is better to make efforts to eliminate Legionella spp. in water supplies.

Methimazole is a drug widely used in hyperthyroidism. However, life-threatening hepatotoxicity has been associated with its clinical use. No protective agent has been found to be effective against methimazole-induced hepatotoxicity yet. Hence, the capacity of organosulfur compounds to protect rat hepatocytes against cytotoxic effects of methimazole and its proposed toxic metabolite, N-methylthiourea was evaluated. Hepatocytes were prepared by the method of collagenase enzyme perfusion via portal vein. Cells were treated with different concentrations of methimazole, N-methylthiourea, and organosulfur chemicals. Cell death, protein carbonylation, reactive oxygen species formation, lipid peroxidation, and mitochondrial depolarization were assessed as toxicity markers and the role of organosulfurs administration on them was investigated. Methimazole caused a decrease in cellular glutathione content, mitochondrial membrane potential (ΔΨm) collapse, and protein carbonylation. In addition, an increase in reactive oxygen species (ROS) formation and lipid peroxidation was observed. Treating hepatocytes with N-methylthiourea caused a reduction in hepatocytes glutathione reservoirs and an elevation in carbonylated proteins, but no significant ROS formation, lipid peroxidation, or mitochondrial depolarization was observed. N-acetyl cysteine, allylmercaptan, and diallyldisulfide attenuated cell death and prevented ROS formation and lipid peroxidation caused by methimazole. Furthermore, organosulfur compounds diminished methimazole-induced mitochondrial damage and reduced the carbonylated proteins. In addition, these chemicals showed protective effects against cell death and protein carbonylation induced by methimazole metabolite. Organosulfur chemicals extend their protective effects against methimazole-induced toxicity by attenuating oxidative stress caused by this drug and preventing the adverse effects of methimazole and/or its metabolite (s) on subcellular components such as mitochondria.

Many studies have been showed transfer of aflatoxins, toxins produced by Aspergillus flvaus and Aspergillus parasiticus fungi, into milk. These toxins are transferred into the milk through digestive system by eating contaminated food. Due to the toxicity of these materials, it seems that it has side effects on the growth of mammary cells. Therefore, the present work aimed to investigate possible toxic effects of aflatoxin B1 (AFB1) on bovine mammary epithelial cells in monolayer and three-dimensional cultures. Specimens of the mammary tissue of bovine were sized out in size 2×2 cm in slaughterhouse. After disinfection and washing in sterile PBS, primary cell culture was performed by enzymatic digestion of tissue with collagenase. When proper numbers of cells were achieved in monolayer culture, cells were seeded in a 24-well culture plate for three-dimensional (3D) culture in Matrigel matrix. After 21 days of 3D culture and reaching the required number of cells, the concentrations of 15, 25 and 35 μL of AFB1 were added to the culture in quadruplicate and incubated for 8 hours. Cellular cytotoxicity was examined using standard colorimetric assay and finally, any change in the morphology of the cells was studied by microscopic technique. Microscopic investigations showed necrosis of the AFB1-exposed cells compared to the control cells. Also, bovine mammary epithelial cells were significantly affected by AFB1 in dose and time dependent manner in cell viability assays. According to the results, it seems that AFB1 can induce cytotoxicity and necrosis in bovine mammary epithelial cells.

Thermal analysis (TGA, DTG and DTA) and differential scanning calorimetry (DSC) have been used to study the thermal behavior of terazosin hydrochloride (TER). Thermogravimetric analysis (TGA/DTG), differential thermal analysis (DTA) and differential scanning calorimetry (DSC) were used to determine the thermal behavior and purity of the used drug. Thermodynamic parameters such as activation energy (E*), enthalpy (H*), entropy (S*) and Gibbs free energy change of the decomposition (G*) were calculated using different kinetic models. The purity of the used drug was determined by differential scanning calorimetry (99.97%) and specialized official method (99.85%) indicating to satisfactory values of the degree of purity. Thermal analysis technique gave satisfactory results to obtain quality control parameters such as melting point (273 ºC), water content (7.49%) and ash content (zero) in comparison to what were obtained using official (272 ºC), (8.0%) and (0.02%) for melting point, water content and ash content, respectively. Thermal analysis justifies its application in quality control of pharmaceutical compounds due to its simplicity, sensitivity and low operational costs. DSC data indicated that the degree of purity of terazosin hydrochloride is similar to that found by official method.

The aim of this article is to synthesize, characterize and evaluate the antimicrobial activity of certain novel 3-methyl-5-oxo-4-(phenyl hydrazono)-4,5-dihydro-pyrazol-1-yl]-acetic acid N|-(4-substituted thiazol-2-yl)-hydrazides. The synthesized compounds were characterized by elemental analysis and IR, NMR and mass spectral data. The antimicrobial activity of novel compounds was evaluated by broth dilution method. XVe, XVf and XVg have shown better antibacterial activity than other compounds of the series. XVa, XVc, XVd and XVe have shown better antifungal activity than the other compounds of the series. All compounds were found to exhibit fair degree of antimicrobial activity.

The aim of this study was to assess the effects of zinc supplementation on anthropometric measures, improving lipid profile biomarkers, and fasting blood glucose level in obese people. This randomized, double- blind clinical trial was carried out on 60 obese participants in the 18-45 age range for one month. The participants were randomly divided into the intervention group, who received 30 mg/d zinc gluconate, and the placebo group who received 30mg/d starch. Anthropometric measurements (body mass index (BMI), weight and waist circumference) were recorded before and at the end of study. Lipid profile biomarkers and fasting blood glucose were determined using enzymatic procedure. Analysis of Covariance (ANCOVA) test was run to compare the post-treatment values of the two groups, and t-test was conducted to compare within group changes. Serum zinc concentration was increased significantly in intervention group (p=0.024). BMI and body weight was significantly decreased (p=0.030 and p=0.020, respectively). Lipid profile biomarkers and fating blood glucose did not change significantly but triglyceride level was significantly decreased (p=0.006) in the intervention group. The obtained results indicate that zinc supplementation improves BMI, body weight, and triglyceride concentration without considerable effects on lipid profile and glucose level. Zinc can be suggested as a suitable supplementation therapy for obese people, but more studies are needed to verify the results.

As a typical Chinese herbal medicine, Radix Glycyrrhizae (RG) possesses various pharmacological effects involved in antioxidant ability. However, its antioxidant has not been explored so far. The aim of the study was to investigate its antioxidant ability, then further discuss the antioxidant mechanism. RG was extracted by ethanol to obtain ethanolic extract of Radix Glycyrrhizae (ERG). ERG was then determined by various antioxidant methods, including DNA damage assay, DPPH assay, ABTS assay, Fe3+-reducing assay and Cu2+-reducing assay. Finally, the contents of total phenolics and total flavonoids were analyzed by spectrophotometric methods. Our results revealed that ERG could effectively protect against hydroxyl-induced DNA damage (IC50 517.28±26.61μg/mL). In addition, ERG could scavenge DPPH· radical (IC50165.18±6.48μg/mL) and ABTS+• radical (IC507.46±0.07μg/mL), reduce Fe3+ (IC50 97.23±2.88 μg/mL) and Cu2+ (IC50 59.21±0.18 μg/mL). Chemical analysis demonstrated that the contents of total phenolics and flavonoids in ERG were 111.48±0.88 and 218.26±8.57 mg quercetin/g, respectively. Radix Glycyrrhizae can effectively protect against hydroxyl-induced DNA damage. One mechanism of protective effect may be radical-scavenging which is via donating hydrogen atom (H·), donating electron (e). Its antioxidant ability can be mainly attributed to the flavonoids or total phenolics.

As a typical Chinese herbal medicine, Citri reticulatae pericarpium (CRP) possesses various pharmacological effects involved in antioxidant ability. However, its antioxidant effects have not been reported yet. The objective of this work was to investigate its antioxidant ability, then further discuss the antioxidant mechanism. CRP was extracted by ethanol to obtain ethanol extract of Citri reticulatae pericarpium (ECRP). ECRP was then measured by various antioxidant methods, including DNA damage assay, DPPH assay, ABTS assay, Fe3+-reducing assay and Cu2+-reducing assay. Finally, the content of total flavonoids was analyzed by spectrophotometric method. Our results revealed that ECRP could effectively protect against hydroxyl-induced DNA damage (IC50 944.47±147.74 μg/mL). In addition, it could also scavenge DPPH· radical (IC50349.67±1.91 μg/mL) and ABTS+• radical (IC5011.33±0.10 μg/mL), reduce Fe3+ (IC50 140.95±2.15 μg/mL) and Cu2+ (IC50 70.46±1.77 μg/mL). Chemical analysis demonstrated that the content of total flavonoids in ECRP was 198.29±12.24 mg quercetin/g. Citri reticulatae pericarpium can effectively protect against hydroxyl-induced DNA damage. One mechanism of protective effect may be radical-scavenging which is via donating hydrogen atom (H·), donating electron (e). Its antioxidant ability can be mainly attributed to the flavonoids, especially hesperidin and narirutin.

A simple RP-HPLC method has been developed and validated for the determination of almotriptan malate (ATM) in bulk and tablets. Chromatographic separation of ATM was achieved by using a Thermo Scientific C18 column. A Mobile phase containing a mixture of methanol, water and acetic acid (4:8:0.1 v/v) was pumped at the flow rate of 1 mL/min. Detection was performed at 227 nm. According to ICH guidelines, the method was validated. The calibration curve was linear in the concentration range 5–60 μg/mL for the ATM with regression coefficient 0.9999. The method was precise with RSD <1.2%. Excellent recoveries of 99.60 - 100.80% proved the accuracy of the method. The limits of detection and quantification were found to be 0.025 and 0.075 μg/mL, respectively. The method was successfully applied for the quantification of ATM in tablets with acceptable accuracy and precision.

To achieve simultaneous imaging and therapy potentials, targeted fluoromagnetic nanoparticles were synthesized and examined in human breast cancer MCF-7 cells. Fe3O4 nanoparticles (NPs) were synthesized through thermal decomposition of Fe(acac)3. Then, magnetic nanoparticles (MNPs) modified by dopamine-poly ethylene glycol (PEG)-NH2; finally, half equivalent fluorescein isothiocyanate (FITC) and half equivalent folic acid were conjugated to one equivalent of it. The presence of Fe3O4-DPA-PEG-FA/FITC in the folate receptor (FR) positive MCF-7 cells was determined via fluorescent microscopy to monitor the cellular interaction of MNPs. FT-IR spectra of final compound confirmed existence of fluorescein on folic acid grafted MNPs. The Fe3O4-DPA-PEG-FA/FITC NPs, which displayed a size rang about 30-35 nm using scanning electron microscopy (SEM) and transmission electron microscopy (TEM), were able to actively recognize the FR-positive MCF-7 cells, but not the FR-negative A549 cells. The uniform nano-sized Fe3O4-DPA-PEG-FA/FITC NPs displayed great potential as theranostics and can be used for targeted imaging of various tumors that overexpress FR.

Urinary tract infection (UTI) is the most common nosocomial infection among hospitalized patients. Meanwhile, most frequent infections involving enterococci affect the urinary tract. The aims of this study were to investigate the susceptibility pattern of isolated enterococci from UTI and the prevalence of virulence genes. The study used enterococci isolated from urinary tract infections obtained from 3 university teaching hospitals in Northwest Iran. The antimicrobial susceptibility of the strains was determined using the disc diffusion method. Multiplex PCR was performed for the detection of genus- species specific targets, and potential virulence genes. Of 188 enterococcal isolates, 138 (73.4%) and 50 (26.6%) were Enterococcus faecalis and E. faecium, respectively. Antibiotic susceptibility testing showed high resistance to amikacin (86.2%), rifampicin (86.2%) and erythromycin (73.9%), irrespective of species. In total, 68.1% were positive for gelE, and 57.4%, 53.2%, 56.4%, and 52.1% of isolates were positive for cpd, asa1, ace, and esp, respectively. The study revealed that most of UTI isolates were multidrug resistance against the antibiotics tested and antibiotic resistance was more common among E. faecium isolates than E. faecalis. A significant correlation was found between UTI and the presence of gelE among E. faecalis strains (p < 0.001).

Progressive loss of dopaminergic neurons of the substantia nigra pars compacta (SNc) in Parkinson’s disease (PD) leads to impairment of motor skills. Several evidences show that the role of serotonergic system in regulation of normal movement is pivotal and mediates via 5-HT1A receptors. Our previous study has shown that fluoxetine in acute injections able to attenuate catalepsy in 6-hydroxydopamine (6-OHDA)-lesioned rats. Since drugs are used chronically in clinic, in this study we attempted to evaluate effect of chronic administration of fluoxetine on 6-OHDA-induced catalepsy. Catalepsy was induced by unilateral infusion of 6-OHDA (8 μg/2 μl/rat) into the central region of SNc and assayed by using bar-test. Fluoxetine (1, 2.5, 5 and 10 mg/kg) was injected intraperitonealy (ip) for 10 days and its anti-cataleptic effect was assessed at the 10th day. Fluoxetine in high doses (5 and 10 mg/kg) worsened 6-OHDA-induced catalepsy while it had anti-cataleptic effect at the dose of 1mg/kg. The anti-cataleptic effect of fluoxetine (1mg/kg) was reversed by co-administration with NAN-190 (0.5 mg/kg, ip), as a5-HT1Areceptor antagonist. According to the results it can be concluded that fluoxetine has anti-cataleptic effect in parkinsonian rats only at low doses, whereas at higher doses it worsens catalepsy. It’s anti-cataleptic effect is exerted through affecting on 5-HT1Areceptors. However, at high doses other mechanisms may be involved. Further clinical studies are needed to prove it’s possible clinical application as an adjuvant therapy in reducing catalepsy of PD.

Fumaria parviflora Lam (Fumariaceae) has been used in traditional medicine in the treatment of several diseases such as diabetes. The present work was designed to evaluate the hypoglycaemic effects of methanolic extract (ME) of F. parviflora in normal and streptozotocin-induced diabetic rats. The rats used were allocated in six (I, II, III, IV, V and VI) experimental groups (n=5). Group I rats served as ‘normal control’ animals received distilled water and group II rats served as ‘diabetic control’ animals. Diabetes mellitus was induced in groups II, V and VI rats by intraperitoneal single injection of streptozotocin (STZ, 55 mg kg-1). Group V and VI rats were addi-tionally treated with ME (150 mg kg-1 day-1 and 250 mg kg-1 day-1, i.p. respectively) 24 hour post STZ injection, for seven consecutive days. Groups III and IV rats received only ME 150 mg kg-1 day-1 and 250 mg kg-1 day-1, i.p. respectively for seven days. The levels of blood glucose were determined using a Glucometer. Administra-tion of F. parviflora extract showed a potent glucose lowering effect only on streptozo-tocin (STZ) induced diabetic rats below 100 mg/dl (P<0.001). However, no significant differences in the blood glucose levels were recorded between diabetic rats received 125 or 250 mg/kg of plant extracts. The findings of the study indicated that F. parviflora has significant hypoglycemic effect on STZ-induced diabetic rats with no effects on blood glucose levels of normal rats.

Monoclonal antibodies or specific antibodies are now an essential tool of biomedical research and are of great commercial and medical value. The purpose of this study was to produce large scale of monoclonal antibody against CD34 in order to diagnostic application in leukemia and purification of human hematopoietic stem/progenitor cells. For large scale production of monoclonal antibody, hybridoma cells that produce monoclonal antibody against human CD34 were injected into the peritoneum of the Balb/c mice which have previously been primed with 0.5 ml Pristane. 5 ml ascitic fluid was harvested from each mouse in two times. Evaluation of mAb titration was assessed by ELISA method. The ascitic fluid was examined for class and subclasses by ELISA mouse mAb isotyping Kit. mAb was purified from ascitic fluid by affinity chromatography on Protein A-Sepharose. Purity of monoclonal antibody was monitored by SDS -PAGE and the purified monoclonal antibody was conjugated with FITC. Monoclonal antibodies with high specificity and sensitivity against human CD34 by hybridoma technology were prepared. The subclass of antibody was IgG1 and its light chain was kappa. The conjugated monoclonal antibody could be a useful tool for isolation, purification and characterization of human hematopoietic stem cells.

The aim of this study was to design and formulation of potassium citrate effervescent tablet for reduction of calcium oxalate and urate kidney stones in patients suffering from kidney stones. In this study, 13 formulations were prepared from potassium citrate and effervescent base in different concentration. The flowability of powders and granules was studied. Then effervescent tablets were prepared by direct compression, fusion and wet granulation methods. The prepared tablets were evaluated for hardness, friability, effervescent time, pH, content uniformity. To amend taste of formulations, different flavoring agents were used and then panel test was done by using Latin Square method by 30 volunteers. Formulations obtained from direct compression and fusion methods had good flow but low hardness. Wet granulation improves flowability and other physicochemical properties such as acceptable hardness, effervescence time ≤3 minutes, pH<6, friability < 1%, water percentage < 0.5% and accurate content uniformity. In panel test, both of combination flavors; (orange - lemon) and (strawberry - raspberry) had good acceptability. The prepared tablets by wet granulation method using PVP solution had more tablet hardness. It is a reproducible process and suitable to produce granules that are compressed into effervescent tablets due to larger agglomerates.

This article makes an attempt to synthesize certain compounds containing thiazole and imidazole moieties and screen for the antimicrobial properties. The novel compounds synthesized were characterized by elemental analysis, IR and 1HNMR spectral data. The antimicrobial activity of novel compounds was evaluated by cup plate method. The compound p-t showed more antibacterial activity than that of the standard. p-hp and p-as showed considerable antibacterial activity. p-t demonstrated higher antifungal activity than that of the standard while p-hp and p-as showed considerable antifungal activity. The antimicrobial activity studies were conducted on certain selected bacteria and fungi. In each case antimicrobial activity of the compounds was compared with that of standards. p-t, p-hp, p-np, p-cp, p-ts and p-as showed considerable antimicrobial activity.

Ambrisentan (ABS) is an antihypertensive drug used in the treatment of pulmonary atrial hypertension. The survey of literature for ABS revealed only two spectrophotometric methods for its quantification. The reported methods lack the sensitivity. This study is aimed at developing two sensitive extractive spectrophotometric methods for the determination of ABS in bulk and in tablets. The proposed methods are based on the formation of colored chloroform extractable ion-pair complexes of ABS with methylene blue (MB method) and safranine O (SO method) in buffered solution at pH 9.8. The extracted complexes showed maximum absorbance at 525 and 515 nm for methylene blue and safranine O, respectively. In both the methods, the calibration curve was linear from 1–15 μg mL−1 of drug. Apparent molar absorpitivities were 1.7911 x 105, 2.3272 x 105 L mol−1 cm−1; Sandell’s sensitivities were 0.0215, 0.0162 μg cm-2; LOD were 0.182, 0.175 μg mL−1; LOQ were 0.551, 0.531 μg mL−1 for methods MB and SO, respectively. The relative standard deviation and percent recovery ranged from 0.206–1.310% and 99.0–101.5%, respectively. The results demonstrate that the proposed methods are sensitive, precise, accurate and inexpensive. These methods can easily be used for the assay of ABS in quality control laboratories.

The study is aimed at finding new antibiotic therapy for aquaculture due to potential of bacteria to develop resistance to the existing therapies. Use of large quantities of synthetic antibiotics in aquaculture thus has the potential to be detrimental to fish health, to the environment and wildlife and to human health. Antimicrobial potential of volatile oil and fractions of chloroform extract of Oreganum vulgare was evaluated alone and in the presence of standard antimicrobials against common fish pathogens by disc-diffusion, agar well assay and two fold microdilution method by nanodrop spectrophotometric method. The best results were represented by volatile oil followed by phenolic fraction by disc-diffusion, agar well and microdilution assays (Minimum inhibitory concentration). By the interaction studies, it was observed that the volatile oil and phenolic fraction were able to inhibit the pathogens at very low concentration compared to standard drugs. The fractional inhibitory concentration index (FICI) was calculated and volatile oil and phenolic fractions were found to be synergistic against Pseudomonas fluorescens and Candida albicans. The experimental data suggests the use of volatile oil and phenolic fraction in combination with standard antimicrobials to maintain healthy aquaculture with lesser adverse effects as compared to synthetic antibiotic therapy.

Pseudomonas aeruginosa and methicillin-resistant Staphylococcus aureus (MRSA) cause difficulties in the management of skin and soft tissue infections and have led to morbidity and mortality in hospital-acquired infections especially in susceptible individuals, those who are generally sick or immunosuppressed. Currently approaches in antibacterial agents offer opportunities to manage the trouble using novel anti-infection systems. Therefore, nanotechnology, a most promising field for generating new applications in medicine, has introduced a most prominent nanoproduct named as nanosilver that revealed excellent antimicrobial activity against some of the hazardous infections. Also cathelicidin peptides which are a part of native immune defense system in the skin and epithelia exhibit excellent antimicrobial activity against some of these perilous infections.