Research in Pharmaceutical Sciences (17355362)(2)
Background and purpose: Alzheimer's disease (AD) is a neurodegenerative disease specified by chronic and irreversible destruction of neurons. This study aimed to evaluate the effects of different extracts (aqueous, hydroalcoholic, hexane, and ethyl acetate) and manna of Echinops cephalotes (EC) on impaired cognitive function induced by scopolamine in mice. EC is shown to have anti-cholinesterase-butyrylcholinesterase activities. Experimental approach: In this study, aqueous and hydroalcoholic extracts, hexane and ethyl acetate fractions of EC (25, 50, 100 mg/kg, i.p.), and the manna (25, 50, 100 mg/kg, gavage) were administered for 14 days alongside scopolamine (0.7 mg/kg, i.p.). Rivastigmine (reference drug) was administered for 2 weeks i.p. Mice were tested for their memory function using two behavioral models, object recognition test (ORT) and passive avoidance test (PAT). Findings/Results: Administration of scopolamine significantly impaired memory function in both behavioral models. In the PAT model, all extracts at 50 and 100 mg/kg significantly reversed the effect of memory destruction caused by scopolamine. At a lower dose of 25 mg/kg, however, none of the extracts were able to significantly change the step-through latency time. In the ORT model, however, administration of all extracts at 50 and 100 mg/kg, significantly increased the recognition index. Only the manna and the aqueous extract at 25 mg/kg were able to reverse scopolamine-induced memory impairment. Conclusions and implications: These results suggest that all forms of EC extracts improve memory impairment induced by scopolamine comparably to rivastigmine. Whether the effects are sustained over a longer period remains to be tested in future work. © 2024 Copyright:
Golnari, M.,
Bahrami, N.,
Milanian, Z.,
Rabbani khorasgani, M.,
Asadollahi, M.A.,
Shafiei, R.,
Fatemi, S.S. Scientific Reports (20452322)14(1)
Despite the current use of some Bacillus spp. as probiotics, looking for and introducing new efficient and safe potential probiotic strains is one of the most important topics in both microbiology and food industry. This study aimed to isolate, identify, and evaluate the probiotic characteristics and safety of some Bacillus spp. from natural sources. Thirty-six spore-forming, Gram-positive, and catalase-positive Bacillus isolates were identified in 54 samples of soil, feces and dairy products. Bacterial identification was performed using 16S rDNA sequencing. To evaluate the probiotic potential of isolates, the resistance of bacterial cells to simulated gastrointestinal tract (GIT) conditions, the presence of enterotoxin genes, their susceptibility to antibiotics, antimicrobial and hemolytic activities and biochemical profiles were investigated. The results revealed that eight sporulating Bacillus spp. isolates fulfilled all tested probiotic criteria. They showed a high growth rate, non-hemolytic and lecithinase activity, and resistance to simulated GIT conditions. These strains exhibited broad-spectrum antibacterial activity against pathogenic bacteria. In addition, they did not exhibit antibacterial resistance to the 12 tested antibiotics. The results of this study suggest that these isolates can be considered as candidates for functional foods and as safe additives to improve diet quality. © 2024, The Author(s).
Research in Pharmaceutical Sciences (17355362)(5)
Background and purpose: Enzyme engineering is the process of raising enzyme efficiency and activity by altering amino acid sequences. Kallikrein 6 (KLK6) enzyme is a secreted serine protease involved in a variety of physiological and pathological activities. The increased expression of KLK6 plays a key role in various diseases. Instability and spontaneous activation and deactivation are major challenges in the study of this enzyme. This study aimed to create a stable pro-KLK6 enzyme by enzyme engineering, designing a specific cleavage site for enterokinase, and using Pichia pastoris GS115 as a host cell. Then, recombinant pro-KLK6 was used to introduce a novel inhibitor for it. Experimental approach: An engineered pro-KLK6 gene was cloned into the pPICZα A expression vector. Then, it was expressed in P. pastoris GS115 and purified by Ni-NTA chromatography. An inactive engineered pro-KLK6 gene was cleaved by enterokinase and converted to an active KLK6. The KLK6 enzyme activity and its kinetic parameters were measured using N-benzoyl-L-arginine ethyl ester (BAEE) substrates. Findings/Results: The secretory form of the pro-KLK6 was expressed at about 11 mg/L in P. pastoris (GS115). Before activation with enterokinase, pro-KLK6 was inactive and did not activate spontaneously. The kinetic parameters, including K m and V max, were estimated at 113.59 μM and 0.432 μM/s, respectively. Conclusion and implications: A stable pro-KLK6 enzyme was produced using P. pastoris (GS115) as the host cell and a specific cleavage site for enterokinase. Additionally, this study assessed the kinetic parameters of the KLK6 enzyme using the BAEE substrate for the first time. © 2023 Wolters Kluwer Medknow Publications. All rights reserved.
Scientific Reports (20452322)13(1)
Recently Bacillus spp. has gained much attention as potential probiotics due to the production of resistant cells. So, this research is purposeful for evaluation of probiotic characteristics of Bacillus isolates from camel milk as a suitable source for growth and isolation of microorganisms that can be candidate to be used as probiotic. First, forty-eight colonies were screened by using morphological and biochemical analysis. Among the isolates, two of them were recognized as Bacillussubtilis CM1 and CM2 by partial 16SrRNA sequencing that, probiotic potentials of them were evaluated. Both of them, in the preliminary safety screening, were found negative for hemolysis and lecithinase activity. Also, in vitro characteristics such as acid, bile salts and artificial gastric juice resistant, cell surface hydrophobicity, auto-aggregation, antioxidant characteristics, and adherent capability to HT-29 cells were determined for them approximately in the range of other probiotic strains. Two strains were susceptible to various antibiotics and enterotoxigenic activities were not detected by PCR which means isolated Bacillus strains could be classified as safe. Altogether, results demonstrate that Bacillus CM1 and CM2 strains could have the potential of consideration as probiotics, however more extensive in vitro/vivo studies are needed. © 2023, The Author(s).
Research in Pharmaceutical Sciences (17355362)(3)
Background and purpose: Properties of Alzheimer's disease, can be caused by several reasons and there is no definite treatment for it. We aimed to study the effect of the hydroalcoholic extract, methanolic and n-hexane fractions of brown algae Sargassum angustifolium on memory impairment in mice and rats. Experimental approach: Hydroalcoholic extract (25, 50, 100, 200 mg/kg), methanolic (20 and 40 mg/kg) and n-hexane (40 and 60 mg/kg) fractions of S. angustifolium were administered for 21 days intraperitoneally before scopolamine injection (2 mg/kg) on day 21. Rivastigmine was administered for 3 weeks intraperitoneally as well. Then, cognitive function was evaluated by three behavioral tests: passive avoidance, object recognition, and the Morris Water Maze test. Findings/Results: Scopolamine induced memory impairment and rivastigmine significantly reversed the memory dysfunction in all three tests. Hydroalcoholic extract and methanolic fraction significantly reversed scopolamine-induced memory impairment in passive avoidance by 64% and 55% and enhanced the recognition index in the object recognition test. In the Morris water maze test probe trial and training session, on days 3 and 4, the hydroalcoholic extract showed a significant decrease in time spent in the target quadrant and path length, respectively. Also, hydroalcoholic extract and methanolic fraction decreased escape latency time in training sessions on days 3 and 4, by 50% and 31% in comparison to scopolamine. N-hexane fractions had no significant effect on scopolamine-induced cognitive impairment. Conclusion and implications: Although the n-hexane fraction wasn't effective, the administration of hydroalcoholic extract and the methanolic fraction of S. angustifolium enhanced scopolamine-induced memory impairment. © 2023 Wolters Kluwer Medknow Publications. All rights reserved.
Food Chemistry: X (25901575)19
The chemical structure and bioactivity of ultrasonic-assisted alkaline extracted polysaccharides of date seed (DSP) and date flesh (DFP) were investigated. In addition, a crossover clinical trial was conducted to evaluate the effects of 28 days of date seed powder and date flesh consumption on blood biomarkers. Xylose (72.2 %) and galactose (41.6 %) were the most abundant monosaccharides in DSP and DFP, also DFP had a higher uronic acid content (12.16 ± 2.13 g/100 g) compared to DSP (5.57 ± 1.2 g/100 g). DSP had higher proliferation and antibacterial effects compared to DFP and inulin. Bifidobacterium animalis produced a higher short-chain fatty acid concentration during fermentation of DSP (66.98 ± 4.33 mM) and DFP (58.58 ± 5.57 mM) than inulin (19.68 ± 3.73 mM). Date seed powder could significantly reduce C-reactive proteins and triglycerides and increase red blood cell count (p < 0.05). DSP showed considerable prebiotic capability, antibacterial activity, and health-promoting effect; therefore, it could be considered for further investigation as nutraceuticals. © 2023 The Authors
Iranian Journal Of Veterinary Research, Shiraz University (17281997)24(1)pp. 65-73
Background: Species of the Bacillus genus have a long history of use in biotechnology. Some Bacillus strains have recently been identified for food applications and industrial as safe bacteria, which mostly have been recognized as probiotic strains. Aims: The primary purpose of the current study was to evaluate the probiotic characteristics of Bacillus subtilis strains isolated and identified from the goat milk samples. Methods: After sampling from 40 goat milk and cultivation, suspected colonies were subjected to biochemical and molecular identification. Then, the confirmed isolate was assessed for in vitro probiotic tests, including hemolysis and lecithinase properties, bile salt, acid, and artificial gastric juice resistance, antioxidant activity, antibiotics susceptibility, enterotoxin genes detection, and attachment capacity to the HT-29 cells. Results: Among 11 suspected isolates evaluated, only one isolate was identified as B. subtilis. In vitro tests for this strain showed similar results to other probiotic strains. The B. subtilis strain was susceptible to various antibiotics. The enterotoxin genes were not detected based on PCR assay. Concerning its probiotic characteristics assessment, especially tolerance to bile salts and acidic conditions, the Bacillus strain could have the potential to consider as a probiotic. Conclusion: Goat milk can be recommended as a source of Bacillus isolates. Also, the isolated strain showed high adaptability to the gastrointestinal environment, relatively equal percentages of adhesion properties, and some safety aspects, having the potential to be considered as an appropriate probiotic. © 2023 Shiraz University. All rights reserved.
Journal of Food Measurement and Characterization (21934126)(4)
Polysaccharides are polymers of sugars linked by glycosidic bonds. They have many nutritional benefits, including prebiotic effects, anti-inflammation, and gut microbiota modulation. Molecular structure investigation helps to understand polysaccharides’ functional, conformational properties and biological activities, facilitating their applications in food and nutraceuticals products. Date fruit (Phoenix dactylifera L.) is an ancient plant cultivated in tropical and subtropical areas. The most abundant bioactive compounds of date fruit are polysaccharides. Several investigations reported that date polysaccharides are biologically active compounds. Date flesh contains around 14% polysaccharide; it constitutes 80% of date seed weight (dry based). Date flesh polysaccharides consist of a particular type of (1→3)-β-D-glucopyranosyl residues with (1→6)-linked branched saccharide residues. Date seed also contains xylan as the primary hemicellulose polymer. Xylan structurally consists of β-1,4-linked D-xylose as the main string with different side chains like L-arabinose, D-galactose, and acetyl groups. The structural analysis of polysaccharides is possible through several specialized techniques. Molecular weight-based separation, monosaccharide composition, linkage patterns, anomeric configuration, and sequences of monosaccharides are the essential properties to be characterized in these investigations. The current review collects comprehensive information about date polysaccharides’ chemical and nutritional properties. © 2022, The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.
Pharmaceutical Development and Technology (10837450)(3)
In this study, a novel wafer based on Hydroxypropyl methylcellulose (HPMC) was prepared as a wound dressing for the simultaneous delivery of phenytoin (PT) and insulin; evaluation of the cutaneous wound repair property was performed too. Due to its low water solubility, PT was encapsulated in polymeric micelles (PM) by the film hydration method at different polymer/drug ratios and characterized in terms of particle size (PS), polydispersity index (PdI), zeta potential (ZP), drug loading (DL) %, entrapment efficiency (EE) %, and drug release. Then, the optimized PT loaded PM (PT-PM) was embedded in the wafers prepared from the HPMC polymer, alone or in combination with Carbopol 940 (CB) and xanthan gum (XG). This wafer also contained a fixed amount of insulin (PT-PM-Insulin-wafer). The obtained wafers were evaluated in terms of morphology, water uptake ability, porosity, bioadhesion and hardness features. Finally, the efficacy of the PT-PM-Insulin-wafer was assessed in full-thickness excision wound models. The optimized PT-PM showed the PS of 84.05 ± 1.80 nm, PdI of 0.28 ± 0.22, ZP of −3.38 ± 0.26 mV, DL of 15.63 ± 0.01%, EE of 92.66 ± 0.08%, and the release efficiency of 59.95 ± 0.03%. The results obtained from the XRD studies of PT-PM also demonstrated the transition of the crystalline nature of the PT to the amorphous form, while FTIR studies showed some intermolecular interaction of PT and the Soluplus® copolymer chain. It was also found that the incorporation of XG into HPMC wafers influenced the microstructure, thus increasing the porosity, water uptake ability and bioadhesion. Compared with other groups, the PT-PM-Insulin-wafer group showed the enhancement of wound closure through increasing collagen deposition and re-epithelialization. The present study, therefore, revealed that the PT-PM-Insulin-wafer group might have very promising applications for wound healing. © 2022 Informa UK Limited, trading as Taylor & Francis Group.
Journal of Isfahan Medical School (10277595)(667)
Background: Neuroprotective effects of L-carnitine on damage caused by fetal exposure to ethanol have been demonstrated. The purpose of this study was to evaluate the effects of different doses of L-carnitine at different stages of pregnancy on the memory of two month old offsprings whose mothers consumed alcohol during pregnancy. Methods: For the purpose of this study, 55 pregnant female rats were randomly divided into 11 groups of 5, including control group, which received drinking water, and the ethanol group which received drinking water containing 10 % (w/v) ethanol. L-Carnitine treated group received L-carnitine in doses of 50, 100, or 150 mg/kg in addition to the ethanol, in the first 10 days or the second 10 days of pregnancy. For modeling the third trimester of pregnancy, doses of L-carnitine were gavaged to the rat neonates for 10 days. After two months of birth, the memories of offsprings were evaluated using Passive Avoidance Task (PAT) and Novel Object Recognition Test (ORT). Findings: Administration of L-carnitine significantly increased discrimination of the ORT index and the escape latency of entry to dark compartment in PAT by two month old offspring compared to the ethanol group. The mentioned indices were significantly increased at a dose of 150 mg/kg compared to the dose of 50 mg/kg throughout all pregnancy periods. These indices were significantly different when l-carnitine was administered in the second 10 days of pregnancy compared to any other periods. Conclusion: The administration of L-carnitine during pregnancy can prevent alcohol-induced memory impairment in rat offsprings. This effect was more significant at higher doses of L-carnitine. The best response was seen when L-carnitine was administered to mothers in the second 10 days of pregnancy. © 2022 Isfahan University of Medical Sciences(IUMS). All rights reserved.
Research in Pharmaceutical Sciences (17355362)(5)
Background and purpose: Alzheimer's disease is a progressive brain disorder that is thought to be triggered via disruption of cholinergic neurons and enhanced oxidative stress. Therefore, antioxidant phytochemicals with the ability to fortify cholinergic function should help in preventing the progress of the disease. This study aimed at evaluating the combinational effects of two popular herbs one with anticholinesterase activity namely Piper nigrum and the other with antioxidant capacity, Cinnamomum zeylanicum. Experimental approach: In this study, P. nigrum extract (PN) (50, 100 mg/kg, ip) and C. zeylanicum extract (CZ) (100, 200, 400 mg/kg, ip) and their combinations were administered for 8 days before the injection of scopolamine (1 mg/kg, ip). Mice were then tested for their memory using two behavioral models, namely the object recognition test and the passive avoidance task. Findings/Results: Administration of scopolamine significantly impaired memory performance in both memory paradigms. In the passive avoidance test (PAT) model, PN at doses up to 100 mg/kg and CZ at doses up to 400 mg/kg did not significantly alter the memory impairment induced by scopolamine. The combination of these two plant extracts did not change the PAT parameters. In the object recognition test (ORT) model, however, administration of 100 mg/kg CZ alone and a combination of PN (50 mg/kg) with CZ (400 mg/kg), significantly increased the recognition index (P < 0.05). Conclusion and implications: Two plant extracts when administered alone or in combinations affected the memory performance differently in two memory paradigms. In the PAT model, the extracts did not show any memory improvement, in ORT, however, some improvements were observed after plant extracts. © 2021 Wolters Kluwer Medknow Publications. All rights reserved.
Advanced Biomedical Research (22779175)(1)
Background: Cognitive impairment is an unpleasant and progressive mental disorder characterized by learning and memory disabilities. Stress and alcohol are two known environmental factors that increase cognitive impairment. This study was designed to evaluate the relative role of cyclooxygenase 2 in alcohol or stress-induced cognitive impairment. Materials and Methods: Male Wistar rats were randomly divided into groups with six rats in each. The groups included sham, control, alcohol (15% ethanol in drinking water), and restraint stress (restraint 6 h per day). Three separated groups received celecoxib at a dose of 20 mg/kg in addition to those listed above. The treatments continued daily for 28 days. The object recognition task (ORT) and Morris water maze (MWM) are used to evaluate the learning and memory. Results: Alcohol or restrain stress significantly increased the time and distance needed to find the hidden platform in MWM. Furthermore, they decreased the recognition index in ORT compared to the control group. Administration of celecoxib significantly decreased the required time and traveled distance to reach the platform in alcohol-treated animals but not in the stress-exposed rats. Celecoxib also significantly increased the recognition index both in alcohol- or restraint stress-exposed animals. Conclusion: We found that either alcohol or restraint stress impairs memory in rats. In MWM, celecoxib improved the alcohol-induced memory impairment but could not show a reduction in memory deterioration due to restraint stress. In ORT, celecoxib reversed memory impairment due to both alcohol and restraint stress. © 2021 Advanced Biomedical Research.
Research in Pharmaceutical Sciences (17355362)(1)
Background and purpose: Doxorubicin (DOX) is an effective agent for the treatment of many neoplastic diseases. Cardiotoxicity is the major side effect of this drug and limits its use. Vanillic acid (VA) is a pharmaceutical compound from the phenolic acids family. The present study is an attempt to investigate the possible helpful effects of VA against DOX-induced cardiotoxicity in rats. Experimental approach: For induction of cardiotoxicity, male Wistar rats received total of six doses of DOX (2.5 mg/kg i.p.) three times per week from days 14 to 28. Treatment groups received daily oral doses of VA (10, 20, and 40 mg/kg) two weeks before DOX injection and then plus DOX for 2 weeks. At the end of experiment, systolic blood pressure (SBP) and heart rate (HR) were detected using tail-cuff method. Lactate dehydrogenase (LDH), creatine phosphokinase-MB (CK-MB), serum glutamic oxaloacetic transaminase (SGOT), malondialdehyde (MDA), and ferric reducing antioxidant power (FRAP) were measured in serum samples. Troponin-I and toll-like receptor 4 (TLR4) were measured in cardiac tissue. All the measurements processed spectrophotometrically using commercial ELISA kits. Cardiac tissue was also processed for histopathological examination. Findings/Results: Treatment with VA significantly increased SBP compared to the DOX group and restored HR near to the normal level. Administration of VA at all of doses, decreased serum levels of LDH, SGOT, CK-MB, MDA, cardiac troponin-I, cardiac TLR4 and increased FRAP value. Conclusion and implications: These results suggest that VA may exert cardioprotective effects against DOX-induced cardiotoxicity by decreasing oxidative stress and biomarkers of cardiotoxicity, suppression of TLR4 signaling and consequently inflammation pathway. © 2020 Wolters Kluwer Medknow Publications. All rights reserved.
Biotechnologia (08607796)(2)
Human growth hormone (somatropin) is one of the most widely used recombinant proteins that stimulates growth, cell reproduction, and cell regulation in humans. Synthetic production of this protein normally results in low yields and inclusion body formation. To overcome these difficulties, the production of somatropin along with two common signal peptides, namely TorA and SufI, in co-expression with a cytosolic chaperone, GroEL/ES, was evaluated in the present study. The target protein and the two signal sequences (TorA and SufI) were synthe-sized and cloned into an expression plasmid (pET-22) by using Nde l and Xho l endonucleases. The expression vector (pGro7) containing chaperone proteins (GroES/EL) and one of the expression vectors containing the signal sequence (and the target protein) were co-expressed in the BL21 DE3 expression host. The results showed that although some of the expressed proteins exit the cytoplasm and enter the periplasmic space, there is also an accumulation of proteins (probably as inclusion body) inside the cytoplasmic area. Western blot analysis showed that the inclusion of a signal sequence in the cassette containing the target protein could help to secrete the protein in the periplasmic space and culture media when compared with control groups. The result of these ex-periments show that the TAT secretion system promotes transportation of the target protein out of the cyto-plasm. This secretory system completes folding of the protein structure and transfers the mature protein to the periplasmic space. © 2020, Institute of Bioorganic Chemistry. All rights reserved.
Iranian Journal Of Fisheries Sciences (15622916)(3)
Rainbow trout fillet is susceptible to microbial and oxidative spoilage. Therefore, it is essential to use preservatives to extend its shelf life. Date extract has significant antibacterial and antioxidant properties. This research was conducted to study the effect of aqueous date extracts on chemical, microbial and sensory properties of farmed rainbow trout during refrigeration. Total phenolic and flavonoid content were determined through Folin-Ciocalteu and colorimetric method. The first step of study was performed for determination the antimicrobial activity of date extracts against the inherent flora of fish fillet, lactic acid bacteria, Pseudomonas and Enterobacteriaceae by disc diffusion method. In parallel with antimicrobial tests, sensory evaluation was also performed for choosing the best concentration of extracts in order to applying on fish fillets. In the second step of study, fish fillet samples were immersed in date extracts (3% w/v for 5 minute), packaged in zip-bags and stored at 4 °C over a period of 17 days. The extract-free fillet was used as control. The samples were analyzed for microbiological (mesophilic, lactic acid bacteria, Pseudomonas and Enterobacteriaceae counts) and chemical (PV, TBARS and TVB-N) parameters. The 5-point hedonic method was carried out for sensory evaluation by 30 trained panelists. Analyses were conducted at 2 h after preparation and 1, 3, 5, 7, 9, 11, 13, 15 and 17 days of storage. Phenolic and flavonoids contents showed no significant differences between date extracts (p>0.05). Total bacterial counts, Enterobacteriaceae, Pseudomonas, TBARS, PV and TVB-N showed no significant difference in test samples (p>0.05). These parameters were within an acceptable range up to 15 days for test samples while the control samples had a shelf life of 5 days. Samples preserved by Piaroum extract had the longest shelf life while samples preserved by the combination of extracts had the shortest. According to the results, the Piaroum, Zahedi, Mozafati date extracts and their combination could be used as natural preservatives for trout fillet shelf-life extension. © 2020 Iranian Fisheries Research Organization. All rights reserved.
Journal Of Reports In Pharmaceutical Sciences (23221232)(1)
Purpose: In the previous studies, we designed an anticancer immunotoxin containing the catalytic and translocation domains of diphtheria toxin fused to BR2, a buforin II-derived antimicrobial peptide as a cancer-specific cell penetrating peptide, in order to target various cancer cells. The aim of this study was to evaluate the in vitro cytotoxicity of DT386-BR2 against K-562 cells as the most famous cell line for leukemia. Materials and Methods: MTT and flow-cytometry assays were used for determining the cytotoxic effects and cell death mechanism of DT386-BR2, respectively, against K-562 cell line. The recombinant DT386 and synthetic BR2 were used as the negative control in cytotoxicity assay. Results: The results of this study showed a significant reduction in survival of K-562 cells caused by DT386-BR2 as compared with BR2 and DT386 fragments. On the contrary, the flow-cytometry results showed apoptosis induction by DT386-BR2 after 12h in a dose- and time-dependent manner. Conclusion: DT386-BR2 fusion protein can be used for further preclinical studies for determining its pharmacokinetic/pharmacodynamic profiles and evaluating its anticancer efficacy in suitable animal models. © 2020 Wolters Kluwer Medknow Publications. All rights reserved.
Clinical Psychopharmacology and Neuroscience (17381088)(2)
Objective: Alzheimer’s disease is a popular neurodegenerative disorder which is growing in the elderly people. Exposure to environmental pollutant like aluminum could trigger or accelerate its involved mechanisms like tau phosphorylation. The current study will evaluate the effect of alone or co-administration of Citicoline or/and magnesium on the aluminum chloride induced memory impairment. Methods: Male albino mice were randomly divided into different groups (n = 7). Memory impairment was induced via orally administration of 300 mg/kg Aluminum Chloride for 28 days. Based on respective group, animals received 100, 250, 500 mg/kg of Citicoline or 50, 100, 150 mg/kg of Magnesium sulfate (MgSO4), intraperitoneally. In co-administration, 50 mg/kg of MgSO4 injected concomitantly with 100, 250, or 500 mg/kg of Citicoline. Rivastigmine (2 mg/kg intraperitoneally) was used as a positive control. Memory was evaluated using the Object Recognition Task (ORT) and Passive Avoidance Test (PAT). Results: The studied doses of Citicoline or MgSO4 when administered individually showed significant increase in the discrimination index in ORT and latency time in the PAT compared to the Aluminium chloride (AlCl3) treated group. Concomitant injection of 50 mg/kg MgSO4 with the different doses of Citicoline strongly increased the above indices values in comparison to each alone. Conclusion: The findings show, individual administration of Citicoline or MgSO4 inverted the AlCl3-induced memory impairment in a dose independent manner. The addition of MgSO4 to the Citicoline showed a synergistic effect in the PAT and likely additive effect in the ORT. Copyright © 2020, Korean College of Neuropsychopharmacology
Clinical Psychopharmacology and Neuroscience (17381088)(1)
Objective: Diabetes mellitus is associated with cognitive disorders such as Alzheimer's disease. Studies have shown that citicoline and benfotiamine can improve memory and learning through different mechanism of actions. The aim of this study was to compare the individual effects of benfotiamine (100, 200, 300 mg/kg) and citicoline (50, 100, 250, 500 mg/kg, gavage) and their co-administration on memory impairments in diabetic mice. Methods: Diabetes was induced by a single dose of streptozotocin (STZ, 140 mg/kg, intraperitoneal) and benfotiamine and/or citicoline were administered for three weeks. Memory was evaluated using the object recognition task (ORT) and passive avoidance test (PAT). Results: Results from ORT shows that citicoline at 50, 100, 250, and 500 mg/kg and benfotiamine at 100, 200, and 300 mg/kg and their combination (benfotiamine at 100 mg/kg added to citicoline at 50, 100, and 250 mg/kg) are equally effective in reversing the memory loss induced by STZ (p ＜ 0.001). PAT results demonstrate that citicoline at 100, 250, and 500 mg/kg and benfotiamine at above doses did not improve the latency time when administered separately, but benfotiamine at a fixed dose of 100 mg/kg in the presence of citicoline at 50, 100, and 250 mg/kg increased the latency time and improved memory significantly. Conclusion: In conclusion, in PAT, co-administration of benfotiamine and citicoline was more effective than either alone in improving memory. Regarding ORT, although benfotiamine added to citicoline improved memory notably, the difference between combination therapy and single-drug therapy was not considerable. Copyright © 2020, Korean College of Neuropsychopharmacology
Research in Pharmaceutical Sciences (17355362)(3)
Development of new medicine with fewer deleterious effects and more efficacies for treatment of inflammatory bowel disease is needed. 5-Hydroxytryptamine 3 receptor (5-HT3R) antagonists have exhibited analgesic and anti-inflammatory features in vitro and in vivo. The present study was designed to evaluate the anti-inflammatory effect of alosetron, a 5-HT3R antagonist, on trinitrobenzenesulfonic acid (TNBS)-induced ulcerative colitis in rats. Two h subsequent to induce colitis (intracolonic instillation of TNBS, 50 mg/kg) in male Wistar rats, alosetron (1 mg/kg), dexamethasone (1 mg/kg), meta-chlorophenylbiguanide (mCPBG, a 5-HT3R agonist, 5 mg/kg), or alosetron + mCPBG were administrated intraperitoneally for 6 days. Animals were thereafter sacrificed and the efficacy of drugs was evaluated macroscopically, histologically, and biochemically (myeloperoxidase, tumor necrosis factor-alpha, interleukin-6, and interleukin-1 beta) on distal colon samples. Treatment with alosetron and dexamethasone improved macroscopic and microscopic colonic damages significantly and decreased myeloperoxidase activity and colonic levels of inflammatory cytokines. The profitable effects of alosetron were antagonized by concurrent administration of mCPBG. Our data provided evidence that the protective effects of alosetron on TNBS-induced colitis can be mediated by 5-HT3R. © 2019 Wolters Kluwer Medknow Publications. All rights reserved.
Iranian Journal Of Basic Medical Sciences (20083866)(9)
Objective(s): To address the alarming problem of methicillin-resistant Staphylococcus aureus (MRSA), herein, a marine Streptomyces capable of producing an anti-MRSA compound has been studied. Materials and Methods: Strain MN41 was morphologically and physiologically characterized and then, molecularly identified using 16SrRNA analysis. To produce the bioactive compound in large scale, a kind of submerged liquid fermentation was adopted. The antibacterial agent was purified using a silica gel column followed by a semi-preparative HPLC and the isolated metabolite was identified using mass spectrometry, Nuclear magnetic resonance (NMR) and Fourier-transform infrared (FTIR). Finally, the production process was subjected to a two steps optimization using Plackett-Burman design (PBD) and Response Surface Method (RSM), respectively. In addition, the antitumor activity of the active agent was studied. Results: The purified compound with a molecular weight of 421.2 was identified as a natural pyrrole-derivative. The optimization revealed a significant effect for starch, pH, calcium carbonate and peptone on the production of this anti-MRSA compound and resulted in a 218% increase in the production yield. Conclusion: The isolated pyrrole-derivative showed a remarkable activity against MRSA and also showed some promising anti-tumor activity. © 2019, Mashhad University of Medical Sciences. All rights reserved.
Research in Pharmaceutical Sciences (17355362)(3)
Lippia citriodora is commonly used in Iranian folk medicine for treatment of many disorders. Since there scientific data to prove the anxiolytic properties of this plant in Iran are scarce, we aimed to evaluate the sedative and anxiolytic activity of the leaf extract and essence of L. citriodora in an animal model of anxiety. The extract and the essence used were obtained after maceration and hydro-distillation of the leaves of L. citriodora, respectively. We evaluated the anti-anxiety profile and sedative activity of diazepam (1 mg/kg i.p. as the standard), hydroethanolic extract (200 and 400 mg/kg i.p.) and the essence (10, 15, and 50 mg/kg i.p.) of leaves of L. citriodora using elevated plus-maze and locomotor activity. We also used flumazenil, to find out if the possible effects are mediated through gamma-aminobutyric acid (GABA)/benzodiazepine receptor complex. The results showed that the essence of L. citriodora at a dose of 15 mg/kg is the most effective anxiolytic dose. Interestingly, flumazenil reversed this action of the essence as well as that of diazepam. The extract even at a dose of 400 mg/kg did not show significant anxiolytic effect. In locomotor activity studies, the essence caused sedation to a lesser extent than diazepam. The results suggest that the essence of this plant could be a better candidate for further analysis and fractionation. As the anxiolytic effect of the essence is reversed by flumazenil, it is possible that the GABA receptor could be involved in mediating these effects. © 2018 Herpetologist's League Inc. All rights reserved.
Journal Of Herbmed Pharmacology (23455004)(2)
Introduction: Stachys lavandulifolia is traditionally used for the treatment of anxiety. Our previous study showed that the ethyl acetate fraction of the plant had substantial anxiolytic action in mice. The present study was aimed to investigate the main constituent responsible for the observed effects. Methods: Ethyl acetate extract was fractionated using column chromatography. Yielded fractions (FR1-4) at 50 mg/kg, and diazepam at 0.5 mg/kg were tested on the elevated plus-maze (EPM). Bioactive fraction was subjected to more purification on repeated chromatography columns. The isolated compound was identified based on 1H-NMR, 13C-NMR, and ESI-Mass spectra. Results: In the time spent in open arm, Fr1, and Fr3 did not show any significant effect on mice behavior, Fr2 marginally decreased the percentage of spent time by -8.4%, and Fr4 significant increased in the time spent in the open arms by 15.7%. In the open arm entry number Fr1, and Fr2 did not show any significant effect on mice behavior, Fr3 marginally increased the percentage of open arm entries by 7.9 %, and Fr4 significantly increased the open arm entry by 18.2%. The bioactive fraction (Fr.4) was subjected to more purification. Phytochemical analysis of Fr4 lead to the identification of 4',5,7-trihydroxyflavon (apigenin). Conclusion: Semi-polar sub-fraction of S. lavandulifolia showed anxiolytic effects by increased time spent and the entry numbers in the open arms comparable to diazepam in the EPM model in mice. Bioactivity-guided isolation leads to the characterization of apigenin with flavone structure as its active constituent. Hence, it might be introduced as a new anxiolytic agent. © 2018 Nickan Research Institute.
Annals of Oncology (09237534)
International Journal Of Molecular And Cellular Medicine (22519637)(1)
Considering antimicrobial resistance problem, marine microorganisms with the bioactivity against multi-drug resistant (MDR) pathogens have attracted many scientific interests. To address this issue, a total of 21 marine actinomycetes isolated from the Caspian Sea have been screened out. Primary screening via cross-streak method revealed that 3 strains: MN2, MN39, and MN40 produce antimicrobial agents with wide spectrum activity. In the second step, the potent strains were characterized morphologically, and then identified genetically using 16S rRNA analysis. After that, the bioactivity of the ethyl acetate extracts of liquid culture against some MDR bacteria has been studied using disc diffusion method. Finally, the exoenzymatic activity of the strains, and the anti-vibrio activity of the extracts have been evaluated. The nucleotide sequence of the 16S rRNA gene (1.5 kb) showed that the potent strains belong to the genus Streptomyces. The results of disk diffusion method indicated that among the 3 potent isolates, MN39 and MN2 produce biomolecules with antibacterial activity against MDR bacteria specially methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus (VRE). In addition, potent strains showed remarkable anti-vibrio activity as well as extracellular enzyme production including amylase and protease. The results of this study revealed that the marine actinomycetes isolated from the sediments of Caspian Sea produce biomolecules effective against MDR bacteria, and suggested that these strains deserve to be studied as potential probiotics due to their anti-vibrio activity besides exoenzyme production. © 2018. All Rights Reserved.
International Journal Of Molecular And Cellular Medicine (22519637)(1)
Considering antimicrobial resistance problem, marine microorganisms with the bioactivity against multi-drug resistant (MDR) pathogens have attracted many scientific interests. To address this issue, a total of 21 marine actinomycetes isolated from the Caspian Sea have been screened out. Primary screening via cross-streak method revealed that 3 strains: MN2, MN39, and MN40 produce antimicrobial agents with wide spectrum activity. In the second step, the potent strains were characterized morphologically, and then identified genetically using 16S rRNA analysis. After that, the bioactivity of the ethyl acetate extracts of liquid culture against some MDR bacteria has been studied using disc diffusion method. Finally, the exoenzymatic activity of the strains, and the antivibrio activity of the extracts have been evaluated. The nucleotide sequence of the 16S rRNA gene (1.5 kb) showed that the potent strains belong to the genus Streptomyces. The results of disk diffusion method indicated that among the 3 potent isolates, MN39 and MN2 produce biomolecules with antibacterial activity against MDR bacteria specially methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococcus (VRE). In addition, potent strains showed remarkable antivibrio activity as well as extracellular enzyme production including amylase and protease. The results of this study revealed that the marine actinomycetes isolated from the sediments of Caspian Sea produce biomolecules effective against MDR bacteria, and suggested that these strains deserve to be studied as potential probiotics due to their antivibrio activity besides exoenzyme production. © 2018 Babol University of Medical Sciences.
Research in Pharmaceutical Sciences (17355362)(3)
Memory impairment is one of the greatest concerns when it comes to long-term CNS-affecting drug administration. Drugs like gabapentin, pregabalin and baclofen are administered in a long-term period in conditions such as epilepsy, neuropathic pain, spasticity associated with spinal cord injury or multiple sclerosis. Despite their wide spread use, few data are available on the effects of these drugs on cognitive functions, such as learning memory. In the present study, the effects of long-term administration of gabapentin, pregabalin and baclofen on memory were investigated in a comparative manner. Male Wistar rats received intraperitoneal (i.p.) injection of gabapentin (30 mg/kg), pregabalin (30 mg/kg), baclofen (3 mg/kg), combination of gabapentin/baclofen (30/3 mg/kg) and combination of pregabalin/baclofen (30/3 mg/kg) once a day for 3 weeks respective to their groups. After the end of treatments, rat memories were assessed using the object-recognition task. The discrimination and recognition indices (RI and DI) in the T2 trials were used as the memory indicating factors. The results showed that daily i.p. administrations of pregabalin but not gabapentin or baclofen significantly decreased DI and RI compared to saline group. In combination groups, either gabapentin or pregabalin impaired discrimination between new and familiar objects. Our findings suggested that pregabalin alone or in combination with baclofen significantly caused cognitive deficits.
International Journal of Food Properties (10942912)
Aspergillus flavus detection and the removal of contaminated pistachio are critically important. In this work, an impedimetric electrochemical DNA sensor using gold nanoparticles on the glassy carbon electrode was developed for the sensitive detection of the aflD gene of A. flavus. This biosensor had a linear range of the detection, which was from 1 nM to 10 µM with the detection limit of 0.55 nM, while the best time for hybridization was 4 hr. Results for the detection and reproducibility of the synthetic 21-nucleotide target and the target 76-nucleotide polymerase chain reaction product were the same due to the sensitivity of measurement. In addition, this biosensor can be used to enumerate A. flavus up to 2.5 × 108 spores/mL. © 2017 Taylor & Francis Group, LLC.
Research in Pharmaceutical Sciences (17355362)(2)
Potassium bromide (KBr), an old antiepileptic agent, is illegally used in pharmaceutical or food industries to improve the product appearance. KBr has been proven to influence several pathways which are important in memory formation. Therefore, the present study aimed to evaluate the effect of KBr on spatial working memory using object recognition task (ORT). Rats received a single dose of KBr (50, 100 or 150 mg/kg), per oral, in acute treatment. KBr long term effects were also studied in animals receiving 50 mg/kg/day of KBr for 28 consecutive days. At the end of treatments, animals underwent two trials of ORT, five min each. In the first trial (T1), animals encountered with two identical objects for exploration. After 1 h, the animals were exposed to a familiar and an unfamiliar object (T2). The exploration times for discrimination (D) and recognition (R) as well as the frequency of exploration for any objects were determined. Acute administration of 150 mg/kg of KBr significantly decreased the discrimination and recognition indices (RI and DI) (P < 0.01) compared to the control. However, lower doses failed to influence the animals' performance in the test. In addition, long term administration of KBr remarkably diminished the DI and RI and the frequency of exploration (P < 0.05). The results of this study indicate that acute doses of KBr as high as 150 mg/kg are required to hamper memory function in ORT. However, cognitive impairment occured with lower doses of KBr when the duration of treatment is extended.
Journal of Isfahan Medical School (10277595)(433)
Background: Cancer is one of the greatest health-related problems and due to increasing drug resistance and severe side effects of chemotherapeutic agents, production of targeted anticancer agents with lower side effects is under consideration. Previously, we produced a fusion protein consisted of catalytic and translocation domains of diphtheria toxin (DT386) fused to BR2, a cancer specific cell penetrating peptide. First steps of this study showed selective antiproliferative effects of DT386-BR2 on cancer cells but not on normal cell lines. The aim of the present study was evaluation of its in-vivo non-specific toxicity in healthy mice. Methods: The fusion protein was produced and purified through recombinant DNA technology. Intraperitoneal injections with various concentrations of DT386-BR2 were done in healthy mice for five consecutive days and they observed for 14 days after the last injection inspecting their food consumption, body weight, body temperature, and finally being dead or alive. Negative controls were injected with normal saline solution. Findings: There was not any significant effect on temperature, body weight, and viability of mice received various concentrations of DT386-BR2 (P < 0.05), and increasing the protein concentration did not show any adverse effects on mice. Conclusion: DT386-BR2 can be used for further pre-clinical studies to determine its pharmacokinetics/pharmacodynamics profiles and evaluation of its anticancer efficacy in suitable xenograft animal models. © 2017, Isfahan University of Medical Sciences(IUMS). All rights reserved.
Iranian Journal of Pharmaceutical Research (17350328)
Stachys lavandulifolia Vahl is an herbaceous wild plant native to Iran which is traditionally used in Iranian folk medicine as a mild sedative tea for reducing anxiety and for treatment of gastrointestinal disorders. Our previous study on ethyl acetate extract of S. lavandulifolia proved anti-anxiolytic activity and so the present study was designed to determine chemical components of this biologically active fraction. The extract was prepared using maceration method. Column chromatography and medium pressure liquid chromatography (MPLC) was used respectively to separate the fractions. Finally, some evaluated fractions were used for high pressure liquid (HPLC) and peak shaving recycle technique to achieve more purification. Separated compounds were determined using NMR analysis and mass spectroscopy. Six compounds have been isolated from ethylacetate extract of aerial parts of S. lavandulifolia including four flavonoids (apigenin, kumatakenin, penduletin and 4ʹ, 7-dihyroxy- 3, 5, 6-trimethoxy flavone), a labdan diterpenoid (labda-13-en-8, 15-diol), and an iridoid. © 2017 by School of Pharmacy.
Journal of Medical Microbiology (00222615)(4)
Purpose. The purpose of this study was to investigate New Delhi metallo-β-lactamase (NDM) production among gramnegative bacilli. Methodology. Antibiogram-resistotyping and detection of New Delhi metallo-β-lactamase (NDM) in clinical isolates of Klebsiella pneumoniae, Acinetobacter baumannii and Pseudomonas aeruginosa and comparative evaluation of the diagnostic performance of three phenotypic methods for NDM detection, with PCR considered as the gold standard, were performed. Minimum inhibitory concentration (MIC) of antibiotics against NDM-positive strains using E-tests and clonal relationship analysis using enterobacterial repetitive intergenic consensus (ERIC)-PCR in these strains were determined. Results. The most effective antibiotics against strains of the species K. pneumoniae were Colistin, Chloramphenicol and Tigecycline; against P. aeruginosa were Fosfomycin and Polymyxins, and against A. baumannii were Polymyxins, Ampicillin/ Sulbactam and Minocycline. Overall, 66, 31 and 40 different resistotypes were observed among K. pneumoniae, A. baumannii and P. aeruginosa strains, respectively. The blaNDM-1 gene was detected in 28 (8.5 %) strains of the bacteria investigated. The sensitivities and specificities of the Meropenem-EDTA combined disk test, the meropenem-dipicolinic acid combined disk test and the modified Hodge test methods for NDM detection were 96.43, 55.15; 96.43, 54.85; and 89.29, 35.15, respectively. Additionally, in spite of the low positive predictive values of these tests, their negative predictive values were high. ERIC-PCR results revealed two main clusters in NDM-positive strains of each of the species P. aeruginosa and A. baumannii, and ten main clusters in K. pneumoniae. In all the NDM-positive strains maximum MIC rates (>256) were observed for all beta-lactam antibiotics. Conclusion. There were high levels of antibiotic resistance and a high frequency of multi-drug resistance and extensive-drug resistance profiles, as well as highly prevalent blaNDM-1 genes in the bacteria investigated. © 2017 The Authors.
Iranian Journal of Pharmaceutical Research (17350328)(2)
Promoter methylation is one of the main epigenetic mechanisms that leads to the inactivation of tumor suppressor genes during carcinogenesis. Due to the reversible nature of DNA methylation, many studies have been performed to correct theses epigenetic defects by inhibiting DNA methyltransferases (DNMTs). In this case novel therapeutics especially siRNA oligonucleotides have been used to specifically knock down the DNMTs at mRNA level. Also many studies have focused on transcriptional gene silencing in mammalian cells via siRNA mediated promoter methylation. The present study was designed to assess the role of siRNA mediated promoter methylation in DNMT3B knockdown and alteration of promoter methylation of Cadherin-1 (CDH1), Glutathione S-Transferase Pi 1(GSTP1), and DNMT3B genes in MDA-MB-453 cell line. MDA-MB-453 cells were transfected with siDNMT targeting DNMT3B promoter and harvested at 24 and 48 h post transfection to monitor gene silencing and promoter methylation respectively. DNMT3B expression was monitored by quantitative RT-PCR method. Promoter methylation was quantitatively evaluated using differential high resolution melting analysis. A non-significant 20% reduction in DNMT3B mRNA level was shown only after first transfection with siDNMT, which was not reproducible. Promoter methylation levels of DNMT3B, CDH1, and GSTP1 were detected at about 15%, 70% and 10% respectively, in the MDA-MB-453 cell line, with no significant change after transfection. Our results indicated that siDNMT sequence were not able to affect promoter methylation and silencing of DNMT3B in MDA-MB-453 cells. However, quantitation of methylation confirmed a hypermethylated phenotype at CDH1 and GSTP1 promoters as well as a differential methylation pattern at DNMT3B promoter in breast cancer. © 2017 by School of Pharmacy.
Research in Pharmaceutical Sciences (17355362)(1)
Tramadol hydrochloride, a synthetic opioid, acts via a multiple mechanism of action. Tramadol can potentially change the behavioral phenomena. The present study evaluates the effect of tramadol after single or multiple dose/s on the spatial memory of rat using object recognition task (ORT). Tramadol, 20 mg/kg, was injected intraperitoneally (i.p) as a single dose or once a day for 21 successive days considered as acute or chronic treatment respectively. After treatment, animals underwent two trials in the ORT. In the first trial (T1), animals encountered with two identical objects for exploration in a five-minute period. After 1 h, in the T2 trial, the animals were exposed to a familiar and a nonfamiliar object. The exploration times and frequency of the exploration for any objects were recorded. The results showed that tramadol decreased the exploration times for the nonfamiliar object in the T2 trial when administered either as a single dose (P<0.001) or as the multiple dose (P<0.05) compared to the respective control groups. Both acute and chronic tramadol administration eliminated the different frequency of exploration between the familiar and nonfamiliar objects. Our findings revealed that tramadol impaired memory when administered acutely or chronically. Single dose administration of tramadol showed more destructive effect than multiple doses of tramadol on the memory. The observed data can be explained by the inhibitory effects of tramadol on the wide range of neurotransmitters and receptors including muscarinic, N-methyl D-aspartate, AMPA as well as some second messenger like cAMP and cGMP or its stimulatory effect on the opioid, gama amino butyric acid, dopamine or serotonin in the brain.
Iranian Journal of Pharmaceutical Research (17350328)
Depressive disorders are more common among persons with chronic diseases such as inflammatory bowel disease and anti-inflammatory effect of some antidepressants such as amitriptyline has been reported. Acetic acid colitis was induced in both reserpinised (depressed) and non-reserpinised (normal) rats. Reserpinised groups received reserpine (6 mg/kg, i.p.) one hour prior to colitis induction. Then Amitriptyline (5, 10, 20 mg/kg, i.p.) was administered to separate groups of male Wistar rats. All treatments were carried out two hours after colitis induction and continued daily for four days. Dexamethasone (1 mg/kg) and normal saline (1 mL/kg) were used in reference and control groups, respectively. At day five, animals were euthanized and colonic tissue injuries were assessed macroscopically and pathologically. Myeloperoxidase activity as a marker of neutrophil infiltration was also measured in colonic tissues. Results showed that reserpine (6 mg/kg, i.p.) intensified colitic condition. Compared to control, amitriptyline (10, 20 mg/kg) and dexamethasone significantly decreased weight of colon and ulcer index in normal and reserpine-induced depressed rats. Myeloperoxidase activity and pathological assessments also proved anti-inflammatory effect of amitriptyline. Our results suggest that amitriptyline, a tricyclic antidepressant, could reduce inflammatory and ulcerative injuries of colon both in normal and depressed rats. So among the wide spread anti-depressant drugs, amitriptyline is a good choice to treat depression comorbidities in patients with IBD. © 2016 by School of Pharmacy Shaheed Beheshti University of Medical Sciences and Health Services.
International Journal of Radiation Research (23223243)(3)
Background: Diarrhea is a well-recognized side effect associated with pelvic radiation; however, there is not any effective common treatment for radiation-induced diarrhea. A popular alternative is probiotics, which have been used in several gastrointestinal disorders. Probiotics are live microbial food supplements. Furthermore, honey is a putative nutritional with a variety of health effects, including antibacterial, antioxidant, anti-inflammatory and prebiotic. The present study evaluated the effects of probiotic with or without honey on radiation-induced diarrhea. Materials and Methods: Sixty-seven adult patients with pelvic cancer underwent radiotherapy for four weeks. They randomized to receive probiotic (n = 22), probiotic plus honey (n = 21) or placebo (n = 24) from one week before radiotherapy for five weeks. Diarrhea grade and stool consistency score were recorded weekly according to the Common Toxicity Criteria system and the Bristol scales, respectively. Results: The results showed a decrease in the daily number of bowel movements (p = 0.003 and 0.006), diarrhea grade (p = 0.001 and 0.001) and the need for antidiarrheal medication (p = 0.021 and 0.041) also an increase in the stool consistency (p = 0.004 and 0.005) in patients who either used probiotic or probiotic plus honey (respectively), these were significant in weeks 4 and 5 of treatment. Conclusion: Probiotics with or without honey can reduce the incidence of radiation-induced diarrhea and the need for antidiarrheal medication.
Iranian Journal of Microbiology (20083289)(2)
Background and Objectives: The most prevalent and worldwide oral disease is dental caries that affects a significant proportion of the world population. There are some classical approaches for control, prevention and treatment of this pathologic condition; however, the results are still not completely successful. Therefore new methods are needed for better management of this important challenge. Chitosan is a natural and non-toxic polysaccharide with many biological applications, particularly as an antimicrobial agent. Chitosan nanoparticle is a bioactive and environment friendly material with unique physicochemical properties. The aim of the present study was to investigate the antimicrobial effect of chitosan and nano-chitosan on the most important cariogenic streptococci. Materials and Methods: For evaluation of antimicrobial effect of chitosan and nano-chitosan against oral streptococci broth micro-dilution method was carried out for four bacterial species; Streptococcus mutans, Streptococcus sobrinus, Streptococcus sanguis and Streptococcus salivarius. Also the effect of these materials on adhesion of above bacteria was evaluated. One-way ANOVA and post hoc Tukey test were used for statistical analysis. Results: The MICs of chitosan for S. mutans, S. sanguis, S. salivarius and S. sobrinus were 1.25, 1.25, 0.625 and 0.625 mg/mL, respectively. The MIC of chitosan nanoparticle for S. mutans, S. salivarius and S. sobrinus was 0.625 mg/mL and for S. sanguis was 0.312 mg/mL. Chitosan and chitosan nanoparticles at a concentration of 5 mg/mL also reduced biofilm formation of S. mutans up to 92.5% and 93.4%, respectively. Conclusion: The results of this study supported the use of chitosan and chitosan nanoparticles as antimicrobial agents against cariogenic Streptococci. © 2016, Tehran University of Medical Science. All rights reserved.
Journal of Microbiological Methods (01677012)
The aim of this study was to produce a fusion protein consisting of the catalytic and translocation domains of diphtheria toxin fused to BR2, a cancer specific cell penetrating peptide, and evaluation of its cytotoxic effects for targeted eradication of cancer cells. For this purpose, The DT386-BR2 structure was predicted using Modeller 9.14 and the best predicted model was selected based on the minimum DOPE score. A synthetic gene encoding DT386-BR2 was cloned in pET28a expression vector, expressed and purified by affinity chromatography. SDS-PAGE and Western blotting confirmed the expression of the DT386-BR2 fusion protein by revealing a band of about 47 kDa after the induction of the expression. Finally, the purified protein was subjected to MTT assay for evaluation of its cyto-lethal effects on cancer and normal cell lines. Statistical analysis showed significant reduction in survival percent of HeLa and MCF-7 cancer cells in comparison to negative control (PBS), while the cytotoxic effect was not significant on the normal cells, i.e. HUVEC and HEK 293. The IC50 of DT386-BR2 for HeLa and MCF-7 was about 0.55 and 2.08 μg/ml, respectively. In conclusion, the production and purification of DT386-BR2 fusion protein was successfully achieved and its cytotoxic effects on the studied cancer cell lines was established. The promising cytotoxic effects of this newly constructed fusion protein made it a suitable candidate for targeted therapy of cancer, and further in vitro and in vivo studies on this fusion protein is underway. © 2016 Elsevier B.V.
Research in Pharmaceutical Sciences (17355362)(5)
Lead belongs to the heavy metal group and is considered as an environmental contaminant. Acute or chronic contact to lead can change the physiological function of human organs. One of the most important disorders following the lead exposure is neurotoxicity. Lead neurotoxicity consists of the neurobehavioral disturbances like cognitive impairment. The aim of the current study is to evaluate the possible protective effect of vitamin C (Vit C), vitamin B12 (Vit B12), omega 3 (ω-3), or their combination on the lead-induced memory disorder. Adult wistar rats were orally administered Vit C (120 mg/kg/day) or Vit B12 (1 mg/kg/day) or ω-3 (1000 mg/kg/day) or their combination for 3 weeks in groups of 7 animals each. Then lead acetate (15 mg/kg/day) was injected intraperitoneally for one week to all pretreated animals. The control group received normal saline as a vehicle while the positive control for cognitive impairment received just lead acetate. At the end of treatments animal memories were evaluated in Object Recognition Task. The results showed, although 15 mg/kg lead acetate significantly declines the memory-evaluating parameters, pretreatment with Vit C, Vit B12, ω-3, or their combination considerably inverted the lead induced reduction in discrimination (d2) index (P < 0.001) and recognition (R) index (P < 0.001, P < 0.05, P < 0.05, and P < 0.001, respectively). Our findings indicate while lead acetate impairs spatial memory in rat, administration of Vit C, Vit B12, ω-3, or their combination prior to the lead exposure inhibits the lead induced cognitive loss. There was no remarkable difference in this effect between the used supplements.
Toxin Reviews (15569543)(1-2)
The effect of ω-conotoxin MVIIA (Ziconotide or SNX-111) as an inhibitor of the neuronal N-type calcium channels on the signs of morphine withdrawal was investigated. The animals were rendered to physical dependence by the continuous intraperitoneal (IP) injections of gradually enhancing the doses of morphine (10–50 mg/kg) during 7 days. The withdrawal was induced in animals through the IP administration of naloxone with the dose of 2 mg/kg or it was elicited spontaneously. The single and chronic intrathecal injections of ω-conotoxin following the naloxone-precipitated withdrawal diminished the number of jumpings (p < 0.05). Moreover, both acute and chronic administrations of the drug alleviated the intensity of ptosis, piloerection, teeth chattering, diarrhea, irritability and genital grooming (p < 0.01) during the two models of the experiment. It can be concluded that the class of N-type calcium channel blockers ameliorate the severity of morphine withdrawal syndrome. © 2016 Informa UK Limited, trading as Taylor & Francis Group.
Journal Of Research In Medical Sciences (17351995)(7)
Background: Radiotherapy is frequently used in treatment approaches of pelvic malignancies. Nevertheless, it has some known systemic effects on blood cells and the immune system that possibly results in their susceptibility to infection. Probiotics are live microbial food ingredients that provide a health advantage to the consumer. Honey has prebiotic properties. The aim of this clinical trial was to investigate probable effects of probiotic or probiotics plus honey on blood cell counts and serum IgA levels in patients receiving pelvic radiotherapy. Materials and Methods: Sixty-seven adult patients with pelvic cancer were enrolled. Patients were randomized to receive either: (1) Probiotic capsules (including: Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus bulgaricus, Bifidobacterium breve, Bifidobacterium longum, and Streptococcus thermophiles) (n = 22), (2) probiotic capsules plus honey (n = 21) or (3) placebo capsules (n = 24) all for 6 weeks. Blood and serum samples were collected for one week before radiotherapy and 24-72 h after the end of radiotherapy. Results: White blood cells (WBC), red blood cells (RBC), platelet counts, and serum IgA level were not significantly changed in patients taking probiotic (alone or plus honey) during pelvic radiotherapy. The mean decrease in RBC count was 0.52, 0.18, and 0.23 × 106 cells/μL, WBC count was 2.3, 1.21, and 1.34 × 103 cells/μL and platelet count was, 57.6, 53.3, and 66.35 × 103 cells/μL for the probiotic, probiotic plus honey, and placebo groups, respectively. The mean decrease of serum IgA was 22.53, 29.94, and 40.73 mg/dL for the probiotic, probiotic plus honey, and placebo groups, respectively. Conclusion: The observed nonsignificant effect of probiotics may be in favor of local effects of this product in the gut rather than systemic effects, however, as a trend toward a benefit was indicated, further studies are necessary in order to extract effects of probiotics or probiotic plus honey on hematologic and immunologic parameters in patients receiving pelvic radiotherapy. © 2015 Journal of Research in Medical Sciences.
European Journal of Pharmacology (00142999)
High prevalence of psychological comorbidities such as depression and anxiety in patients with inflammatory bowel disease (IBD) supports the premise that adding an anti-depressant drug with known anti-inflammatory effect to the medical treatment have beneficial effect in the course of the underlying disease. Colitis was induced by intracolonic instillation of 2 ml of 4% v/v acetic acid solution in rats. Anti-colitic effect of fluvoxamine was evaluated in two categories: A: normal rats, B: reserpinized (6 mg/kg, i.p.) depressed rats. In group A, fluvoxamine (2.5, 5, 10 mg/kg, i.p.) was administered 2 h after induction of colitis and in group B: reserpine (6 mg/kg, i.p.) was administered 1 h prior to colitis induction and then fluvoxamine (2.5, 5, 10 mg/kg, i.p.) was administered 2 h after colitis induction. Dexamethasone (1 mg/kg) was used as reference drug. All the treatments continued daily for five days. The effect was assessed on the basis of macroscopic score, biochemical (myeloperoxidase) changes and histopathological studies. Results showed that fluvoxamine (2.5 and 5 mg/kg) and dexamethasone treatment markedly reduced disease severity in both reserpinized and non-reserpinized rats as indicated by reduction in macroscopic and microscopic colonic damages while reserpine adversely exacerbated the colitis damage. Myeloperoxidase activity which was increased following colitis induction was also decreased. The findings of this study elucidate the anti-colitic and anti-inflammatory properties of fluvoxamine and so introduced it as a good candidate to treat depressive symptoms in people comorbid to IBD. © 2014 Elsevier B.V. All rights reserved.
Journal of Isfahan Medical School (10277595)(328)
Only a short time after the discovery of antibiotics by Alexander Fleming, antibiotic-resistant bacteria were reported. Besides the traditional mechanisms of resistance in bacteria, new mechanisms have been described by different researchers. For example, recently a new gene operon of D-alanine-Dserine called VanL that cuased resistant to the antibiotic vancomycin was reported. Other researchers have shown that Klebsiella pneumoniae strains lost purine Ompk36 that have a major role in the antibiotic diffusion into cells, are resistant to carbapenem. In 2011, the first full resistance to tigecycline was reported that the enzyme-linked monooxygenase flavin hydroxyl TetX cause deactivation of antibiotics. In 2012, high resistance to mupricin antibiotic was described that intermediated by a new locus named mupB. In addition, in 2013, it was showed that some strains of Enterococcus faecalis can move daptomycin antibiotic from its target to other places and so inhibit its activity. Removal of whole target gene (PBP3) of ceftazidime in Burkholderia pseudomallei was reported in 2012. In other hand, new role of riboswitch for resistance to aminoglycoside antibiotics reported in 2013. In addition, recent studies have indicated a relationship between SOS response and quorum sensing with antibiotic resistance and new antibiotic resistance mechanisms in bacterial biofilm and persister cells are known. In this review, recent studies have been conducted to identify these new mechanisms. © 2015, Isfahan University of Medical Sciences(IUMS). All rights reserved.
Journal of Isfahan Medical School (10277595)(355)
Background: Leptin deficiency or dysfunction in leptin receptor (ObR) signaling may tend to resistance to autoimmune diseases. On the other hand, leptin receptors exist on many cells and therefore blocking all of them will probably result in unfavorable effects. Targeted ObR blocking on specific immune cells with a leptin antagonist such as taFv (Tandem single chain fragment variable or Tandem scFv) may be advantageous for patients with autoimmune diseases. This project aimed to optimize the condition for large scale production of such molecule and to test its effect. Methods: The cloned taFv gene was sub-cloned from pAB1 to pET32a vector. The taFv fragment existence in pET32a vector was confirmed via polymerase chain reaction (PCR) method using T7 primers. Dot blotting was recruited to detect protein expression. Optimization experiments were carried out and assayed using enzyme-linked immunosorbent assay (ELISA). Finally, the functional activity was evaluated via flow cytometry. Findings: The result of PCR confirmed integration of taFv 2300 bp gene fragment in pET32a vector. Dot blotting confirmed taFv higher expression in pET32a vector compared to previous vector. It was found that media containing sorbitol, Escherichia coli BL21 strain, IPTG 0.05 mM and 18° C temperatures were resulted in higher production of protein levels. Based on flow cytometry, taFv was able to attach to 20% of lymphocytes. Conclusion: pET32a vector with pel B fragment is suitable for secretory overexpression. Production of taFv could be enhanced via optimizing media and culture conditions. © 2015, Isfahan University of Medical Sciences(IUMS). All rights reserved.
Jundishapur Journal Of Natural Pharmaceutical Products (17357780)(1)
Background: Reteplase is a mutant version of t-PA (tissue plasminogen activator) with prolonged half-life. In the present study, E. coli Top 10 bacteria were utilized in the production of reteplase, which is the nonglycosylated active domain of t-PA. Reteplase gene was ligated into pBAD/gIII plasmid which, allows secretion of this protein in periplasmic space. It would allow the correct formation of disulfide bonds in protein structure. Objectives: This study aimed at expression of reteplase in optimum condition. In this study, the reteplase gene was cloned and expressed in Escherichia coli top 10 as a suitable host cell and its expression was optimized. Materials and Methods: The recombinant plasmid, pET15b/reteplase was digested by NcoI and BamHI restriction enzymes; while pBAD/gIIIA vector was digested by NcoI and BglII. Then the insert and vector were ligated and used for transformation of E. coli Top10 cells by heat shock method. Overnight culture of transformed bacteria was induced by L-arabinose in various concentrations (0.2, 0.02, 0.002, and 0.0002%) and at various temperatures. Results: The obtained recombinant plasmid was sequenced to confirm the presence and correct framing of reteplase gene regarding the expression of reteplase. Maximum production of this enzyme was obtained under the following condition: 0.0002% L-arabinose at 37°C for 2 hours incubation. The purified protein was detected on SDS-PAGE (sodium dodecyl sulfate Polyacrylamide gel electrophoresis) as a 66 kDa band. The concentration of t-PA standard was 1 unit which is equal to 12 μg/mL. The enzymatic activity of samples was measured as 0.8 units compared to the standards. Conclusions: Reteplase was expressed in E. coli Top 10 after activation of pBAD/gIIIA promoter region by arabinose and optimized. © 2015, School of Pharmacy, Ahvaz Jundishapur University of Medical Sciences.
International Journal of Diabetes in Developing Countries (09733930)(4)
Immunology Letters (01652478)(2)
Different commensal bacteria employed as probiotics have been shown to be endowed with immunomodulatory properties and to actively interact with antigen presenting cells, such as dendritic cells and macrophages. In particular, different strains of probiotic bacteria may induce the secretion of a discrete cytokine profile able to induce divergent T cell polarization. Here, we briefly review current knowledge regarding the effects of different species and strains of probiotic bacteria on T cell polarization. Given that the loss of intestinal homeostasis is frequently associated with an aberrant T cell polarization profile, a comprehensive knowledge of the immunomodulatory potential of these bacteria is crucial for their employment in the management of human immune-mediated pathologies, such as allergies or inflammatory bowel diseases. © 2015 European Federation of Immunological Societies.
Avicenna Journal Of Medical Biotechnology (20082835)(3)
Background: One of the most important producers of high quality industrial enzymes is the Gram-positive bacterium, Bacillus subtilis (B. Subtilis). One major limitation that hinders the wide application of B. subtilis is the secretion of high levels of extracellular proteases which degrade the secreted foreign proteins. In this study, homologus recombination technique was used to knock out its protease gene, aprE. Methods: The internal segment of the pro-sequence of aprE gene of B. subtilis 168 with a length of 80 bps and its complementary sequence were synthesized and ligated into pUB110 at EcoR1 and XbaI restriction sites. Competent cells of B. subtilis 168 were prepared and transformed by electroporation using Bio Rad gene pulser as explained in the methods section. Transformants carrying the recombinant plasmid were selected for resistance to neomycin. The success of homologous recombination was checked by PCR amplification of the neomycin gene which was part of the vector and did not exist in the genome of B. subtilis 168. The protease activity was measured using the Protease Fluorescent Detection Kit based on the proteolytic hydrolysis of fluorescein isothiocyanate (FITC)-labeled casein-substrate. Results: The results demonstrated that aprE gene would not be able to produce further active subtilisin E. The reduction of protease activity also confirmed the efficacy of the induced mutation in this gene. Conclusion: It will therefore be a major challenge for future research to identify and modulate quality control systems of B. subtilis which limit the production of high quality protease- sensitive products such as lipase. © 2014, Avicenna Journal of Medical Biotechnology. All rights reserved.
World Journal of Microbiology and Biotechnology (09593993)(3)
The present study deals with isolation and characterization of copper oxide nanoparticles resistant Pseudomonas strains that were isolated from the soil collected from mining and refining sites of Sarcheshmeh copper mine in the Kerman Province of Iran. The three isolates were selected based on high level of copper oxide nanoparticles (CuO NPs) resistance. The isolates were authentically identified as Pseudomonas fluorescens CuO-1, Pseudomonas fluorescens CuO-2 and Pseudomonas sp. CuO-3 by morphological, biochemical and 16S rDNA gene sequencing analysis. The growth pattern of these isolates with all the studied CuO NPs concentrations was similar to that of control (without CuO NPs) indicating that CuO NPs would not affect the growth of isolated strains. A reduction in the amount of exopolysaccharides was observed after CuO NPs-P. fluorescens CuO-1 culture supernatant interaction. The Fourier transform infrared spectroscopy (FT-IR) peaks for the exopolysaccharides extracted from the bacterial culture supernatant and the interacted CuO NPs were almost similar. The exopolysaccharide capping of the CuO NPs was confirmed by FT-IR and X-ray diffraction analysis. The study of bacterial exopolysaccharides capped CuO NPs with E. coli PTCC 1338 and S. aureus PTCC 1113 showed less toxicity compared to uncoated CuO NPs. Our study suggests that the capping of nanoparticles by bacterially produced exopolysaccharides serve as the probable mechanism of tolerance. © 2013 Springer Science+Business Media Dordrecht.
Genetics in the Third Millennium (24237159)(4)
Probiotics are microbial food additives that improve the intestinal microbial balance through beneficial effects on the host. Recent studies have proved the benefits of probiotics in maintaining the health and strength of the body and fighting against intestinal and other diseases. Extensive use of probiotics is common but for better utilization in industry and medicine, recombination in probiotics is taken into consideration. Genetic manipulation in probiotics can improve carbohydrate fermentation, increase the production of specific metabolites, produce or increase the activity of enzymes (proteolytic, Alpha-Amylase, Beta-Galactosidase and antioxidant enzymes) or increase the capacity of the production of beneficial substances such as bacteriocins, vitamins, and anti-inflammatory substances. On the other hand, some lactic acid bacteria have expressive capabilities for overexpression; therefore, they could be used as a cellular factory for the production of proteins. In the present study, several original papers are reviewed and the most important findings on recombinant probiotics are classified and presented. The results indicated that recombinant probiotics could be used to improve the properties of probiotics (anti-adhesive lining, resistance to acid, etc), production of vaccines, drug transfer, reduction of allergy, immune system modulation, and creation of more effective metabolic pathways. © 2014, Iranian Neurogenetics Society. All rights reserved.
Research in Pharmaceutical Sciences (17355362)(6)
We have previously evaluated the effect of nimodipine, L-type calcium channel blocker, on memory loss during spontaneous morphine withdrawal. In the present study the effect of nimodipine on memory loss in naloxone-induced morphine withdrawal mice was investigated. Mice were made dependent by increasing doses of morphine for three days. Object recognition task that was used for evaluation of memory performance comprised of three sections: 15 min habitation, 12 min first trial and 5 min test trial. Naloxone was injected 3 h after the administration of the last dose of morphine. Recognition index was evaluated 20 min after naloxone injection. Nimodipine was administrated in repeated form (1, 5 and 10 mg/kg) with daily doses of morphine or as a single injection (5 and 10 mg/kg) on the last day. Both acute and repeated treatments with nimodipine prevented the memory impairment in naloxone-induced morphine withdrawal mice (P<0.05 comparison of acute and repeated treatment data with their corresponding control values). Corticosterone concentration was significantly increased in the brain and blood of the mice during withdrawal. Pretreatment with nimodipine, however, decreased the corticosterone concentration in both brain and blood. The present study showed that nimodipine prevents intense memory loss following naloxoneinduced morphine withdrawal.
Avicenna Journal of Medical Biotechnology (20084625)(3)
Background: Production of tissue Plasminogen Activator protein (t-PA) in pro-karyotes systems has many problems such as the lack of active protein pro-duction, multiple purification steps, and renaturation process which has been shown to be costly and time-consuming. Methods: In this study, reteplase which is the nonglycosylated active domain of t-PA was used to transform TOP10 Escherichia coli (E. coli) bacteria to resolve some of the above mentioned problems. Reteplase cDNA was ligated into pBAD/gIII plasmid which allowed secretion of this protein into the periplasmic space and would allow the correct formation of disulfide bonds in protein structure. The presence of reteplase cDNA in pBAD/gIII plasmid was con-firmed by restriction digestion and sequencing. After induction of the expres-sion of this protein by adding 0.0002% L-Arabinose to the medium, the pro-teins in periplasmic space as well as the inclusion bodies formed inside the cell were extracted. Subsequently, these proteins were purified and detected by Western blot method. Results: Our results showed that the amount of reteplase extracted from peri-plasmic space was much lower than the extracted inclusion bodies and large quantities of the recombinant protein were present as inclusion bodies. There-fore, it was more efficient to use inclusion body extraction method for protein isolation and purification. Conclusion: We produced active reteplase after its expression in E. coli TOP10 and isolation of inclusion bodies produced the best results for purification and extraction of this protein. © 2013, Avicenna Journal of Medical Biotechnology.
Research in Pharmaceutical Sciences (17359414)(1)
Apoptosis has a critical role in the pathogenesis of bleomycin induced-pulmonary fibrosis. The severity of fibrosis varies among different strains of mice. Recent studies have indicated that expression of apoptotic regulatory genes may be specific in different cell types in various strains. In this study, bleomycin-induced pulmonary apoptosis in NMRI (Naval Medical Research Institute, USA) albino mice were compared with C57BL/6 black mice. Pulmonary fibrosis induced by single intratracheal administration of bleomycin (3 U/kg). Control mice were instilled with the same volume of saline. After 2 weeks, fibrotic responses were studied by biochemical measurement of collagen deposition and histological examination of pathological lung changes. Apoptosis was detected and quantitated by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Bleomycin significantly (P<0.05) increased lung collagen content and also induced fibrotic histological changes in both strains. Apoptosis was detected in the bronchiolar and alveolar epithelial cells after bleomycin instillation. TUNEL-positive alveolar epithelial cells in bleomycintreated lungs of C57BL/6 and NMRI mice (19.5% ± 2.7 and 17% ± 2.0, respectively) were significantly (P<0.05) higher than that of saline-treated lungs (1.5% ± 0.5) with no significant difference between two strains of mice (P>0.05). Despite some murine strain variation in the expression of apoptotic regulatory genes in bleomycin-induced pulmonary fibrosis, the results of the present study revealed no significant differences in alveolar epithelial apoptosis between NMRI and C57BL/6 black mice. However, these results confirm the role of apoptosis in the pathogenesis of pulmonary fibrosis and suitability of both strains as experimental models of lung fibrosis.
Comparative Clinical Pathology (1618565X)(5)
Brucellosis is a zoonotic disease which is characterized by abortion and reduced fertility in many species. Camel brucellosis is caused by Brucella abortus and Brucella melitensis. To investigate sensitive methods in the detection of camel brucellosis, PCR was used to overcome the limitations of serology and culture methods. Three hundred ten camels were examined for brucellosis infection using serological tests (RBPT, mRB, Wright, and 2-ME). In addition, 100 serological tested cases (39 mRB positive and 61 mRB negative) were analyzed with both bacteriological (lymph node culture on Brucella agar supplemented with antibiotics) and PCR (nested-PCR on sera and blood samples) methods. The nested-PCR was genus-specific and amplified the 16S rDNA locus. Six out of 310 (1.94 %) of the examined camels were positive using the serological tests, whereas, no bacteria was isolated from lymph tissues. Nested-PCR was positive in six and nine individuals in sera and blood samples, respectively. The genus-specific nested-PCR assay on blood samples detected a higher number of camel brucellosis compared with serological and classical culture methods. These results have identified a sensitive PCR method which could be used as a complement test for the detection of brucellosis in live camels with the lowest risk of infection to laboratory workers. © 2012 Springer-Verlag London Limited.
Preparative Biochemistry and Biotechnology (15322297)(7)
Reteplase is the recombinant type of tissue plasminogen activator variant. In this study, preplasmic and cytoplasmic (as inclusion body: IBs) production and activity of recombinant reteplase in E. coli were investigated and compared using a pET system (pET22b and pET15b). The cDNA of reteplase was cloned by polymerase chain reaction (PCR) amplification, sequenced, inserted into the vector pET 22b and pET15b, and expressed using isopropyl β-D-1- thiogalactopyranoside (IPTG). The recombinant plasmid was expressed in the form of inclusion body in pET 15b and in periplasmic space in pET22b. The obtained results of inclusion body extraction from recombinant pET22b (rpET22b) and recombinant pET15b (rpET15b) plasmids using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed a band of ∼39 kD. However, the obtained results of periplasmic space extraction from rpET22b plasmid showed a very weak band, while cytoplasmic expression of reteplase (pET15b) produced a strong protein band confirmed with Western blotting. Consequently, our results demonstrated that the cytoplasmic expression system is efficient for the production of reteplase protein in prokaryote systems and a high amount of reteplase was obtained from the expressed proteins in the form of IBs. The obtained activity of rpET15b plasmid showed a higher enzyme absorbance in comparison to rpET22b plasmid. This suggests rpET15b as an appropriate candidate for reteplase production. © 2013 Copyright Taylor and Francis Group, LLC.
Journal Of Medicinal Plants (2717204X)(45)
Background: Insomnia, which is difficulty in initiating and maintaining sleep, is a very common experience for many people. Considering the increasing interest in medicinal plants in the past decade, many plants such as Coriandrum sativum, Salvia leriifolia, Salvia reuterana and Stachys lavanduli folia have been used in Iranian traditional medicine to abate insomnia. Objective: The present study was designed to investigate hypnotic effect of Salvia reuterana on male mice. Methods: Ethanolic extract of S. reuterana was prepared. Five groups of 6 animals each were pretreated with vehicle, Salvia extract (50, 100 and 250 mg/kg; i.p.) or diazepam (0.5mg/kg; i.p.) 30 minutes before ketamine injection (100 mg/kg, i.p.). Results: The latency and total sleeping times were recorded to determine the hypnotic effect of the extract. The results indicated that ethanolic extract of S. reuterana, reduced the latency time and induced the total sleeping time in a dose dependent manner, compared to saline group. Conclusion: The present study suggests that S. reuterana produces hypnotic effect which can be evaluated clinically.
Introduction: There is a pressing need for research leading to the development of new effective drugs with lower side effects and more efficacy for treating inflammatory bowel disease (IBD). The analgesic and anti-inflammatory properties of 5-Hydroxytryptamine (5-HT)-3 receptor antagonists have been shown in in vivo and in vitro studies. The present study was designed to investigate the effects of tropisetron, a 5-HT3 receptor antagonist, on an immune-based animal model of IBD. Methods: In the present study, the trinitrobenzenesulfonic acid (TNBS) model of colitis in the rat was used. Two hours after induction of colitis in rats, tropisetron (2 mg/kg), dexamethasone (1 mg/kg), meta-chlorophenylbiguanide (mCPBG, 5 mg/kg), a 5-HT3 receptor agonist, or tropisetron + mCPBG were intraperitoneally (i.p.) administrated for 6 days. Animals were then sacrificed; macroscopic, histological, biochemical (myeloperoxidase [MPO]) assessments and ELISA test (tumor necrosis factor-alpha, interleukin-6 and interleukin-1 beta) were performed on distal colon samples. Results: Tropisetron or dexamethasone treatment significantly reduced macroscopic and microscopic colonic damages. In addition, a significant reduction in MPO activity and colonic levels of inflammatory cytokines was seen. The beneficial effects of tropisetron were antagonized by concurrent administration of mCPBG. Conclusion: The present study indicates that the protective effects of tropisetron on TNBS-induced colitis can be mediated by 5-HT3 receptors. © 2013 by Tabriz University of Medical Sciences.
Journal of Isfahan Medical School (10277595)(213)
Background: This study was aimed to clone and express the reteplase complementary deoxyribonucleic acid (cDNA), a thrombolytic agent used for the treatment of acute myocardial infarction and stroke, in Escherichia coli using tac promoter. Methods: Reteplase cDNA was amplified using polymerase chain reaction (PCR) with designed primers. The product was then cloned into pTZ57 plasmid. The cloned DNA was digested out and ligated into pGEX-5x-1 expression vector. The presence of the insert was confirmed by restriction digestion and determination of the nucleotide sequence. By using 0.2, 0.5, and 1 mM isopropyl beta-D thiogalactopyranoside (IPTG), reteplase was induced in escherichia coli TOP10 cells and analyzed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Findings: Electrophoresis of PCR product as well as double digested recombinant pTZ57 plasmid showed a 1068 base pair band of reteplase. SDS-PAGE analysis showed a 60 kilodalton band of protein product induced by IPTG. Conclusion: In the present study, reteplase cDNA was successfully cloned and expressed using tac promoter. This vector was used for the optimization of the expression of reteplase in escherichia coli.
Research in Pharmaceutical Sciences (17359414)(1)
Abrupt cessation of morphine leads to withdrawal signs and cognitive deficits. Endocannabinoid system is activated during withdrawal; therefore, the aim of the present study was to assess the effects of AM281, cannabinoid antagonist/inverse agonist, on memory deficit following spontaneous morphine withdrawal. Cognition was evaluated by using the object recognition task. The novel object recognition task was tested in a square wooden open-field box using objects. The test was consisting of three sections: 15 min exploration, first trial for 12 min and second one for 5 min. In the second trial the difference in exploration between a previously seen object and a novel one, was considered as an index of memory performance (recognition index - RI). Male mice were made dependent by increasing doses of morphine (30-90 mg/kg) subcutaneously twice daily for 3 days. AM281 (0.62, 1.25 and 2.5 mg/kg) were used in chronic form concurrent with morphine i.p. or acutely (2.5, 5 and 10 mg/kg) on the last day. RI was evaluated on the third day 4 h after the last dose of morphine. Chronic administration of AM281 at 2.5 mg/kg improved RI to the 22.1 ± 4.8 and single dose of AM281 at 5 mg/kg improved the memory impairment to the 8.5 ± 4, as compared with vehicle-treated which was 4.8 ± 2.5. The results suggested that administration of AM281 at a dose of 2.5 mg/kg in chronic form and 5 mg/kg in acute dose improved memory.
Evidence-based Complementary and Alternative Medicine (17414288)
The aim of the present study was to assess the effects of chronic and acute treatment of the essential oil (EO) of Kelussia odoratissima Mozaff. on the development of morphine tolerance and dependence in mice. Mice were rendered tolerant to and dependent on morphine by subcutaneous injection of morphine over a period of 5 days. Tolerance was assessed using the tail-pinch test and withdrawal signs of morphine were precipitated by injecting naloxone 2h after the final morphine injection. Repeated injection of the EO of K. odoratissima (5 and 10mg/kg) for 4 days significantly suppressed morphine-withdrawal jumps, a sign of the development of dependence to opiate as assessed by naloxone precipitation withdrawal on day 5 of testing. A single injection (25, 50, 100mg/kg) of the EO on day 5, 1h prior to morphine failed to produce any significant change in morphine withdrawal signs. Neither the acute nor the chronic administration of EO of the K. odoratissima did significantly influence the development of tolerance to the analgesic effect of morphine. Alleviation in morphine signs of withdrawal after chronic injection with K. odoratissima is indicative of reversal of neuronal adaptation that takes place during morphine presence in the brain. © 2012 Mohammed Rabbani et al.
Iranian Journal of Basic Medical Sciences (20083874)(5)
Objective(s) Cannabinoids have been implicated in memory deficit. We examined the effect of AM281, cannabinoid antagonist/inverse agonist in prevention of scopolamine-induced cognitive deficit. Materials and Methods Object recognition task was used to evaluate memory in mice. Exploration time in the first and the second trial was recorded. The differences in exploration between a previously seen object and a novel object in second trial were taken as an index of memory. Scopolamine and AM281 were administrated at the same time, 40 min before second trial in the treatment group. Results Object discrimination was impaired after scopolamine (2 mg/kg; IP) administration. AM281 (2.5, 5 mg/kg; IP) significantly restored object recognition ability in mice treated with scopolamine by 75%. Conclusion This study extends earlier findings, suggesting the interaction of cannabinoid and cholinergic system in memory. Additionally cannabinoid antagonists seem to show variable pharmacological properties.
Inflammatory bowel disease (IBD) is a chronic inflammatory disorder of the intestinal tract whose etiology has not yet been fully elucidated. Available medicines for treatment of IBD are not universally effective and result in marked deleterious effects. This challenge has thus height-ened the need for research in order to adopt new therapeutic approaches for the treatment of IBD. 5-HT 3 receptor antagonists have shown analgesic and anti-inflammatory properties in vitro and in vivo. Our aim was to investigate the effect of ondansetron, 5-HT 3 receptor antagonist, in an im-mune-based animal model of IBD, trinitrobenzene sulfonic acid (TNBS)-induced rat colitis and probable involvement of 5-HT 3 receptors. Two hours after induction of colitis (instillation of 50 mg/kg of TNBS dissolved in 0.25 ml of ethanol 50 % v/v) to male Wistar rats, ondansetron (2 mg/kg), dexamethasone (1 mg/kg), meta-chlorophenylbiguanide (mCPBG, 5 mg/kg), a 5-HT 3 receptor agonist, or ondansetron + mCPBG were administrated intraperitoneally (ip) and continued daily for six days. The animals were sac-rificed and distal colons were assessed macroscopically, histologically and biochemically [myeloperoxidase (MPO), tumor necrosis factor-alpha, interleukin-6 and interleukin-1 beta]. Ondansetron and dexamethasone resulted in a decrease in macroscopic and microscopic colonic damage significantly. In addition a dramatic reduction in MPO activity and colonic levels of in-flammatory cytokines were seen. The protective effects of ondansetron were antagonized by concurrent administration of mCPBG. Our data suggests that the beneficial effects of ondansetron in TNBS-induced colitis could be mediated by 5-HT 3 receptors.
Magnesium Research (19524021)(1)
The present study was conducted to investigate the effect of oral administration of calcium gluconate and magnesium acetate on morphine withdrawal syndrome. Mice were rendered dependent on morphine by subcutaneous injection of increasing doses of morphine. Mice were observed for 30 minutes for the withdrawal signs (jumping or standing events, diarrhea, piloerection, tremor and ptosis). Separate oral administration of magnesium (50, 75 and 100 mg/kg) and calcium (500, 750 and 1,000 mg/kg) significantly decreased the jumping, without affecting standing in animals withdrawn from morphine. Co-administration of magnesium (at a fixed dose of 100 mg/kg) and calcium (at a range of doses from 250 to 1,000 mg/kg) resulted in a significant reduction in jumping and standing events (P<0.05). In a similar fashion, the qualitative signs of withdrawal were also reduced when the above combination of calcium and magnesium was administered. Co-administration of calcium/magnesium at 500/50, 750/75 and 1,000/100 mg/kg significantly reduced the number of jumps in morphine-dependent animals without affecting the number of standing events. This study demonstrates the potential activity of the co-administration calcium and magnesium in preventing the signs associated with morphine withdrawal syndrome.
Gastroenterology Research and Practice (1687630X)
There is a pressing need for research that will lead to the reveal of targets designed to analyse the possible pathways for the treatment of IBD. Because of the probable involvement of serotonin in inflammatory conditions of intestine and the important role of 5HT4 receptors in GI function, the investigation of the role of 5HT4 receptors in the pathogenesis of IBD will be interesting. The aim of this study was to investigate the effects of cisapride, a 5HT4 receptor agonist, in trinitrobenzenesulfonic-acid-(TNBS) induced rat colitis. Two hours subsequent to induction of colitis using TNBS in rats, cisapride (2mg/kg, intraperitoneally (i.p); 4mg/kg, orally (p.o)) and dexamethasone (1mg/kg, i.p; 2mg/kg, p.o) were administrated for 6 days. Animals were thereafter euthanized; macroscopic, histological, and biochemical assessments and ELISA test were carried out on distal colon samples. Our data showed that dexamethasone treatment (i.p, p.o) significantly decreased macroscopic and microscopic damage and also biochemical markers, but there were no significant differences in aforementioned parameters between cisapride (i.p or p.o) and TNBS-treated rats. It can be deduced that because the severity of colitis produced by TNBS is massive (through various pathways), cisapride could not bring about more colitis damages through 5HT4 receptors. Based on the present study further researches are required for investigating the exact roles of 5HT4 receptors in the pathogenesis of ulcerative colitis. Copyright © 2012 Azadeh Motavallian et al.
Journal of Isfahan Medical School (10277595)(193)
Background: The aim of this project was to isolate and purify the highly active recombinant Taq DNA polymerase from the strain of Escherichia coli BL21. This enzyme, with a molecular weight of about 94 kDa, is widely used in polymerase chain reaction (PCR). In PCR, the activity and fidelity of Taq polymerase can significantly influence the results. One of the active regions of Taq polymerase has been suggested to be the O-helix region. In previous studies, an expression vector containing mutated Asn 666 Glu Taq polymerase gene was designed. In order to investigate the effects of this mutation on the function of the enzyme, Taq polymerase needs to be purified first. Methods: In this study, after transformation of competent cells, enzyme expression was induced by isopropyl β D thiogalactopyranoside (IPTG) method. Modified Desai protocol with DNase and RNase, nickel-nitrilotriacetic acid (Ni-NTA) resin, trichloroacetic acid (TCA), and refolding protocols were subsequently used for purification of this enzyme. These protocols were finally compared. Findings: Using Desai protocol resulted in the production of a sharp band in the expected region (94 kDa) and several other visible bands. After further modification of Desai protocol, only the desirable band was observed. In protocols using TCA and Ni-NTA resin, the expected bands were weak. Refolding protocol caused a band in an undesirable region (66 kDa). Conclusion: From the different purification techniques that were used in this study, the modified method of Desai containing RNase and DNase worked best. Addition of TCA can precipitate proteins that had not been affected by heat. Using Ni-NTA resin resulted in elimination of unwanted bands. However, the amount of Taq polymerase was also decreased. The extra band that was observed in refolding protocol was probably due to the presence of proteases that were isolated with inclusion body and could digest Taq polymerase.
Jundishapur Journal Of Microbiology (20083645)(4)
Background: Sugar alcohol erythritol is a non-caloric sweetener, non-cariogenic, and safe for diabetics because of no change to blood glucose and insulin levels after oral administration. Erythritol cannot be degraded by any enzymatic systems and must be eliminated from the blood through the kidney. Objectives: The aim of this study was production and optimization of erythritol from glucose by Yarrowia lipolytica. Materials and methods: Yarrowia lipolytica DSM70562 was cultivated at 30°C in a 250 mL Erlenmeyer containing 50 mL of production medium composed of 200 g/L glucose, 10 g/L yeast extract, 10 mg/L MnSO4.4H2O, and 2 mg/L CuSO4.5H2O. Erythritol was separated from the sugars and other polyols by thin layer chromatography. Total polyols was determined using colorimetric method of Bok and Demain, and erythritol was also eluted from the paper and determined by this colorimetric method. Results: In a batch culture with 200 g/L glucose at pH 5.5, an erythritol producer of Y.lipolytica capable to produce 27.8 g/L erythritol after seven days was selected, corresponding to a 21.52% yield and a productivity of 0.165 g/L/h. Conclusions: In this investigation we optimized the production medium and through altering medium components that resulted in a drastic change in polyol composition. Present study reports the production of erythritol for the first time by a Y. lipolytica strain DSM70562. Due to increasing demand for erythritol as a low caloric sweetener in food industry, its production via biological processes is becoming increasingly important. © 2012 Ahvaz Jundishapur University of Medical Sciences; Published by Kowsar Corp.
Iranian Journal of Pharmaceutical Research (17350328)(1)
The aim of the present study was to synthesis a series of phthalimides based on our previous works and examine their anxiolytic properties. Using a three steps process, phthalimides were prepared from the corresponding di-methyl phthalate derivatives. Phthalic anhydride was nitrated to produce 3-nitrophthalic acid. Ring closer of either 3-nitrophthalic acid or di-methyl phthalate with urea were carried out in reflux condition. Final compounds were prepared by base catalyzed condensation of 4-methylbenzoyl chloride, benzoyl chloride and benzyl chloride with the resulting imides. From the tested compounds, only N-benzoyl 3-nitro-phthalimide was shown to produce anxiolytic activity by increasing the number of entries and time spent in open arms at 10 mg/kg. © 2012 by School of Pharmacy.
Basic and Clinical Pharmacology and Toxicology (17427843)(3)
Morphine withdrawal leads to the activation of endocannabinoid system and cognitive deficits. The aim of this study was to evaluate the effects of AM281, a cannabinoid antagonist/inverse agonist, on memory deficit following naloxone-precipitated morphine withdrawal in mice. Male mice were made dependent by increasing doses of morphine (30-90 mg/kg) twice daily for 3 days. The object recognition task was used to evaluate memory dysfunction. The test comprised three sections: habituation for 15 min., first trial for 12 min. and test trial for 5 min. In this learning paradigm, the difference in exploration between a previously seen object and a new object is taken as an index of memory performance (recognition index). The recognition index was assessed on the third day of morphine treatment by the injection of 0.1 mg/kg naloxone 3 hr after the last dose of morphine. Chronic administration of AM281 at 2.5 mg/kg significantly improved the memory impairment, producing a recognition index of 36.0 ± 3.9 as compared with vehicle-treated data (recognition index = -3.1 ± 8.2%). A single dose of AM281 at 5 mg/kg improved the recognition index from -1.5 ± 3.9% in morphine withdrawal animals to 18.5 ± 11.6%. Concurrent administration of AM281 with morphine proved to be more effective in protecting the animals from losing their memory compared to acute action of AM281. These results indicate that the contribution of the cannabinoid system to memory deficit is attributable to morphine withdrawal. By blocking cannabinoid receptors, AM281 may become useful in preventing memory deficit after morphine withdrawal. © 2012 Nordic Pharmacological Society.
European Journal of Pharmacology (18790712)(1-3)
Effects of the nimodipine, L-type calcium channel antagonist, has been studied on memory loss caused by spontaneous morphine withdrawal in mice. Mice were made dependent by increasing doses of morphine over three days. Memory was evaluated using object recognition task, which is based on tendency of rodents to exploration of new objects. The test was comprised of three sections: 15 min habitation, 12 min first trial and 5 min test trial. Recognition index was evaluated 4 h after the last dose of morphine. Nimodipine was administrated either in chronic form (1, 5 and 10 mg/kg) with daily doses of morphine or it was given as a single injection (5 and 10 mg/kg) on the last day. Nimodipine in both treatment forms prevented the memory impairment following spontaneous morphine withdrawal. Corticosterone concentration was increased in brain and blood of mice during abstinence phase and pretreatment with nimodipine prevented the increase in brain and blood corticosterone concentration. The results show that blockade of L-type calcium channels improves memory deficits caused by morphine withdrawal. This indicates that some kind of treatments, such as nimodipine, administrated over the acute withdrawal phase, can prevent memory deficit during withdrawal. © 2012 Elsevier B.V. All rights reserved.
Research in Pharmaceutical Sciences (17359414)(3)
Trinitrobenzene sulfonic acid (TNBS)-induced colitis is one of the most common methods for studying inflammatory bowel disease in animal models. Several factors may, however, affect its reproducibility, rate of animal mortality, and macroscopic and histopathological outcomes. Our aim was to validate the main contributing factors to this method and compare the effects of different reference drugs upon remission of resultant colon injuries. TNBS was dissolved in 0.25 ml of ethanol (50% v/v) and instilled (25, 50, 100 and 150 mg/kg) intracolonically to the male Wistar rats. After determination of optimum dose of TNBS in male rats and assessment of this dose in female rats, they were treated with reference drugs including dexamethasone [1 mg/kg, intraperitoneally (i.p.) and 2 mg/kg, orally (p.o.)], Asacol (mesalazine, 100 mg/kg, p.o.; 150 mg/kg, enema) and hydrocortisone acetate (20 mg/kg, i.p.; 20 mg/kg, enema) which started 2 h after colitis induction and continued daily for 6 consecutive days. Thereafter, macroscopic and microscopic parameters and clinical features were assessed and compared in different groups. We found that the optimum dose of TNBS for the reproducibility of colonic damage with the least mortality rate was 50 mg/kg. Amongst studied reference drugs, hydrocortisone acetate (i.p.), dexamethasone (i.p. and p.o.) and Asacol (p.o.) significantly diminished the severity of macroscopic and microscopic injuries and could be considered effective for experimental colitis studies in rats. Our findings suggest that optimization of TNBS dose is essential for induction of colitis under the laboratory conditions; and gender exerts no impact upon macroscopic and histological characteristics of TNBS-induced colitis in rats. Furthermore, the enema forms of hydrocortisone and Asacol are not appropriate reference drugs.
Research in Pharmaceutical Sciences (17359414)(2)
The anxiolytic effect of the flower of Echium. amoenum was shown in several experimental studies in mice. The present study was aimed to determine whether tolerance develops to anxiolytic action of E. amoenum in mice. NMRI male mice were injected intraperitoneal with hydroalcoholic extract (12.5, 25 and 50 mg/kg) or saline once each day (8 am) for period of 7 days and then tested on light/dark box model. Anxiolytic effect was determined by light/dark box and elevated plus-maze. According to the results, hydroalcohoic extract of E. amoenum when given both acutely and chronically (7 days) at 25 and 50 mg/kg, significantly increased the time in the illuminated zone. The number of transitions in the light/dark apparatus, however, was not significantly altered by the tested doses of the plant. Diazepam at 0.5 and 1 mg/kg produced anxiolytic effect in both model of anxiety, namely, the light/dark box and elevated plus-maze. No tolerance was developed to the anxiolytic effect of E. amoenum extract after 7 days of treatment. Our results suggest that one week treatment with extract of the E. amoenum does not produce tolerance to its anxiolytic action. Longer period of treatment using implant procedure is probably necessary to cause molecular changes in order to induce tolerance.
OBJECTIVE: The present study aimed to evaluate the sedative and anxiolytic effects of the essential oils and hydroalcoholic extract of Kelussia odoratissima Mozaff. (K. odoratissima) in mice by utilizing an elevated plus maze. The chemical composition of its essential oil was also determined. METHODS: The hydroalcoholic extract or essential oil fraction from this plant were administered intraperitoneally to male mice at various doses 30 min before testing. The anxiolytic and sedative effects were determined by an elevated plus maze and locomotor activity tests, respectively. RESULTS: According to the results, none of the administered doses of hydroalcoholic extract or essential oil fraction of K. odoratissima changed the percentage of the time spent or number of entries into the open arms of the elevated plus maze. In contrast, the cumulative spontaneous locomotor activity of mice treated with the essential oil or hydroalcoholic extract was significantly decreased. Chemical analysis of the essential oil by Gas chromatography- mass spectromentry (GC-MS) showed that 3-butylidene-4,5-dihydrophthalide (85.9%) was the major component. CONCLUSION: These data confirm the sedative properties of K. odoratissima, yet there were no profound anxiolytic effects observed. © 2011 CLINICS.
Genetic Testing and Molecular Biomarkers (19450257)(1-2)
The estrogen receptor β (ERβ) mediates the action of estrogen on metabolism of lipids and lipoprotein. Therefore, its gene is a promising candidate gene for cardiovascular disease. The aim of the present study was to investigate whether the ERβ A1730G polymorphism modifies the metabolic response to hormone replacement therapy (HRT) in postmenopausal women. The population included 60 normolipidemic postmenopausal women with equal numbers of each A1730G genotype followed during a 90-day experimental period. All subjects received oral estrogen together with a progestin therapy during the HRT. ABCA1 gene expression and serum lipid and lipoprotein concentrations were measured at the beginning and end of the HRT trial. At baseline, ABCA1 gene expression, lipid or lipoprotein concentrations were not significantly different among the ERβ A1730G genotype groups. After HRT, however, subjects with GG genotype had a greater increase in ABCA1 gene expression (p=0.002) and a trend toward greater increase in apoA-I (p=0.058) than subjects carrying the A allele. An interaction effect between genotype and HRT effect was observed on ABCA1 gene expression. In conclusion, the positive changes of ABCA1 gene expression and apoA-I were affected by the ERβ A1730G polymorphism in women taking estrogen-progesterone therapy. Copyright 2011, Mary Ann Liebert, Inc.
Gynecological Endocrinology (14730766)(9)
Objective. The ATP binding cassette A1 (ABCA1) is a key participant in the reverse cholesterol process whereby mediates cholesterol efflux directly to HDL particles. The aim of this study was to investigate whether long-term treatment with conventional hormone replacement therapy (HRT) in post-menopausal women could affect their leukocytes ABCA1 expression. Changes in various serum lipids and lipoprotein fractions were also investigated. Material and Methods. A total of 60 non-obese normolipidaemic post-menopausal women treated with oral oestrogen together with progestin therapy for 3 months were selected. Leukocytes ABCA1 gene expression and serum lipid and lipoprotein concentrations were measured at the start and end of the HRT. Results. HRT led to significant increases in HDL cholesterol (P = 0.001) and apoA-I (P = 0.046) and significant decrease in apoB (P = 0.049) and LDL cholesterol (P = 0.022) when compared with the baseline levels. Analysis of leukocytes ABCA1 mRNA showed a significant increase in ABCA1 gene expression after HRT (P = 0.001). There was also a significant inverse association (r = -0.28, P = 0.03) between ABCA1 gene expression and log TG/HDL cholesterol changes related to HRT. Conclusion. The beneficial cardiovascular effects of HRT could be explained, at least in part, by increasing the ABCA1 gene expression. © 2011 Informa UK, Ltd.
Annals of Clinical Biochemistry (00045632)(3)
Research in Pharmaceutical Sciences (17359414)(2)
Reteplase is a segment of tissue plasminogen activator used for the removal of thrombi in blood vessels. In the present study the cloned reteplase gene was used for its expression in competent E. coli. The recombinant plasmid, pET15b/reteplase (rpET-BL21), was transformed into competent E.coli strain BL21 (DE3) cells. Overnight culture of the transformed bacteria was induced by the addition of isopropylthio-ß-Dgalactoside (IPTG) to the final concentrations of 0.25, 0.5, 1 and 1.5 mM. Also, the effects of different temperatures(25, 30, 37 and 39°C), shaking speeds (100, 170 and 190 rpm), and various glucose concentrations (0.25, 0.5, 0.75 and 1 mM) on the expression of reteplase were examined. Samples were analyzed by SDS-PAGE. Maximum amount of protein production was obtained by the addition of 1 mM IPTG at 37°C, 100 rpm of shaking speed in the absence of glucose.
Journal Of Medicinal Plants (2717204X)(39)
Anxiety disorders are amongst the most popular diseases which interfere with normal life. Benzodiazepines are used as a first line of treatment, but difficulties with pharmacotherapy of anxiety disorders such as dependence and low response rate, encourage researchers to find new approaches. From the past, the role of medicinal plants have been a subject of intense interest. In this respect, Citrus aurantium, Coriandrum sativum, Crocus sativus, Echium amoenum, Nepeta persica Boiss, Stachys lavandulifolia and Salix aegyptiaca are widely used by Iranian population. This review summarized the information on Iranian plant species that have been explored for their potential anti-anxiety profile using validated animal models, doses and possible mechanism.
Journal of Complementary and Integrative Medicine (15533840)(1)
The aim of the present study was to evaluate the anxiolytic effects of total extract of flowers of Salix aegyptiaca on the elevated plus-maze (EPM) model of anxiety. The extract of the flower parts of the plant was administered by i.p. and p.o. to male NMRI mice, at various doses. Oral and i.p. administration of the S. aegyptiaca significantly increased the percentage of time spent in the open arms of the EPM at 200 and 100 mg/kg, respectively. The plant extract at 100 and 200 mg/kg significantly decreased the animal's locomotor activity at 10 and 15 min time intervals. These doses, however, did not affect the ketamine-induced sleeping time. The 100 mg/kg dose of the plant extract when given by i.p. route seemed to be the optimal dose in producing the anxiolytic effects. Lack of interaction with ketamine in increasing the sleep time is an advantage over diazepam which is water down by its reduction in locomotor activity. Copyright © 2010 The Berkeley Electronic Press. All rights reserved.
Journal of Pediatric Gastroenterology and Nutrition (02772116)(5)
Objective: The objective of the study was to evaluate the clinical efficacy of oral bovine colostrum in the management of nonorganic failure to thrive (FTT). Materials and Methods: In a randomized clinical trial, 120 children (1-10 years of age) of either sex with mild or moderate nonorganic FTT were divided into 2 groups. Both groups were matched with regard to age, sex, weight, and height. One group (control) received routine treatments for FTT and the other group (case), besides routine treatments, received supplementary bovine colostrum at the dose of 40 mg • kg-1 • day-1 for a 3-month period. Following the initial visit, subsequent visits were completed following 1, 2, and 3 months of supplementation. For quantitative measurements, Waterlow I (height for age) and Gomez (weight for age) indices were used. Results: The mean value of Gomez index in the case group (81.72) was significantly higher than the control group (77.12) at the end of the third month of supplementation (P = 0.003). Such a difference was not reported based on Waterlow I index between the case and control groups (92.91 vs 91.71; P = 0.094). According to Gomez index 12 patients (20%) who received colostrum became healthy at the end of the third month, which was significantly higher than the control group (2 cases, 3.3%); P = 0.006. Conclusions: Bovine colostrum supplementation for a 3-month period is a useful method without any side effects, in addition to known medical and psychological treatments, to increase the weight of children with nonorganic FTT. © Copyright 2010 by European Society for Pediatric Gastroenterology.
INTRODUCTION: We investigated the antianxiety and sedative effects of the essential oil of Ducrosia anethifolia. Boiss. (Apiaceae). METHODS: We used elevated plus maze, spontaneous motor activity and ketamine-induced sleep tests in mice. In addition, the essential oil was analyzed by GC/MS. Twenty compounds were identified, and n-decanal (70.1%) and alpha-pinene (12.4%) constituted the major components. RESULTS: In elevated plus maze, Ducrosia anethifolia essential oil at doses of 25-200 mg/kg increased the percentage of open arm time and entries. Unlike diazepam, ducrosia anethifolia essential oil could not suppress spontaneous motor activity and did not alter ketamine-induced sleep parameters. These results are indicative of antianxiety effect of Ducrosia anethifolia essential oil without sedative effect. © 2010 CLINICS.
Iranian Journal Of Biotechnology (17283043)(1)
The present study was undertaken to screen the soil samples collected in Iran for the presence of the Bacillus subtilis lipase A gene. The bacterial colonies obtained from the collected soil samples were examined by physical appearance, biochemical tests and the polymerase chain reaction (PCR). Only four colonies were identified as putative B. subtilis strains and all contained the lipase A gene. However, the intensities of the DNA bands were different and correlated with the differences obtained from the biochemical tests. Polymorphism of the lipase gene was also determined in samples using SSCP assay. In conclusion, this study demonstrates an easy and reliable method for detection of the lipase gene in B. subtilis strains. Further screening of the soil by this method will enable the detection and identification of industrially more favorable lipases.
Research in Pharmaceutical Sciences (17359414)(1)
Taq DNA polymerase is widely used in laboratories and for this reason many investigators have focused their attention on understanding the role of various regions and amino acids in this enzyme. O-helix is a part of taq polymerase suggested to play an important role in the enzyme fidelity. The influence of Asn666 in this helix on the enzyme function has never been investigated, and therefore by using nested PCR, a portion of taq DNA polymerase gene containing Asn666Glu mutation was amplified. This DNA was digested with Eco RI restriction enzyme to confirm the presence of Asn666Glu mutation. After digesting this product and the wild type taq-pET-15b plasmid with NheI and BamHI restriction enzymes, they were ligated and used for the transformation of E. coli DH5α competent cells. The obtained colonies were screened for the presence of the mutated taq polymerase gene using EcoRI, NdeI and BamHI restriction enzymes. In conclusion, with the use of the obtained recombinant plasmid it is possible to study the role of this amino acid on taq DNA polymerase function.
DARU, Journal of Pharmaceutical Sciences (15608115)(4)
Background and the purpose of the study: Vasopressin type 2 receptor (V2R), a G protein coupled receptor (GPCR), plays an important role in the regulation of renal antidiuretic function. The highly conserved DRH motif is essential for G protein signaling of V2R; however its role especially regarding the histidin residue is not fully understood. Methods: Site directed mutagenesis was performed with replacements of the histidin to isoleucine by using nested polymerase chain reaction. ELISA was performed for receptor expression assay and the adenylyl cyclase activity assay was performed for functional characterization of DRI mutation on V2R signaling.Results and major conclusion: The adenylyl cyclase activity assay in COS-7 cells showed no difference in the amount of cAMP production between the wild type and the mutant V2 receptors. The V2 receptor expression was not changed in the presence of this mutation using ELISA assay. These results suggest that the role of histidin residue is not critical in the V2 receptor function, however further mutagenesis studies are required to define the role of this motif in V2R function.
Comparative Clinical Pathology (1618565X)(3)
The presence of major virulence factors of enterohaemorrhagic Escherichia coli (EHEC; stx1, stx2, eae, Ehly) were determined among isolates from 158 diarrhoeic calves by multiplex polymerase chain reaction (PCR). Strains positive for virulence factors were subjected to serotype specific PCR assays for O157:H7 and O111 antigens. Additionally, serogroups were determined by three monovalent antisera for O26, O111 and O157 somatic antigens and enterohaemolysin production were also shown phenotypically. Thirteen (8. 2%) calves carried strains positive for one or more of the virulence factors tested, and eleven (6. 9%) calves harboured the shiga toxin-producing strains (stx1 or stx2). stx1 was detected in eight (5%) and stx2 in three (1. 9%) calves. eae and Ehly were observed in the same frequency (6. 3%) and were detected in parallel. Of the 13 virulence-positive strains, the predominant genotype was (stx1/eae/Ehly) at 53. 8%. None of the EHEC in this study belonged to O157:H7 or O111 serotypes, but four strains (30. 7%) belonged to the O26 serogroup. The results show the possible role of stx1/eae in calf diarrhoea and the particular importance of O26 EHEC. Calves can also act as a reservoir for EHEC and in the transmission of the disease to humans. © 2009 Springer-Verlag London Limited.
Iranian Biomedical Journal (1028852X)(1-2)
Background: Vasopressin type 2 receptor (V2R) plays an important role in the water reabsorption in the kidney collecting ducts. V2R is a G protein coupled receptor (GPCR) and the triplet of amino acids aspartatearginine- histidine (DRH) in this receptor might significantly influence its activity similar to other GPCR. However, the role of this motif has not been fully confirmed. Therefore, the present study attempted to shed some more light on the role of DRH motif in G protein coupling and V2R function with the use of sitedirected mutagenesis. Methods: Nested PCR using specific primers was used to produce DNA fragments containing aspartate-lysine-isoleucine and aspartate-arginine-tyrosine mutations with replacements of the arginine to lysine and histidine to tyrosine, respectively. After digestion, these inserts were ligated into the pcDNA3 vector and transformation into E. coli HB101 was performed using heat shock method. The obtained colonies were analyzed for the presence and orientation of the inserts using proper restriction enzymes. After transient transfection of COS-7 cells using diethylaminoethyl-dextran method, the adenylyl cyclase activity assay was performed for functional study. The cell surface expression was analyzed by indirect ELISA method. Results: The functional assay indicated that none of these mutations significantly altered cAMP production and cell surface expression of V2R in these cells. Conclusion: Since some substitutions in arginine residue have shown to lead to the inactive V2 receptor, further studies are required to define the role of this residue more precisely. However, it seems that the role of the histidine residue is not critical in the V2 receptor function.
Journal of Biomolecular Techniques (19434731)(2)
Glycosylation of the μ-opioid receptor may play an important role on its function. Using nested PCR, N53Q mutation was prepared in the N-terminal region of the rat μ-opioid receptor cDNA and cloned into the pcDNA3 vector. The plasmids containing the wild-type and mutated receptor cDNA were transfected into the COS-7 cells. Intracellular cAMP was measured in the morphine-treated and untreated transfected cells using an ELISA kit. Plasmid expression was evaluated using X-gal staining. Intracellular concentration of cAMP in the N53Q-mutated cells was not significantly different from the wild-type. The expression of the transfected plasmids was confirmed. Therefore, based on these results, it seems that glycosylation at the N53 site of the rat μ-opioid receptor does not influence the function of this receptor significantly. © 2010 ABRF.
Veterinary Record (00424900)(22)
The virulence properties of Shiga toxin-producing Escherichia coli (STEC) strains isolated from diarrhoeic and non-diarrhoeic calves were compared. The strains were also tested for O157:H7, O111 and O26 serotypes, using PCR and conventional serotyping methods. E coli strains isolated from 297 faecal samples, from 200 diarrhoeic and 97 non-diarrhoeic calves, were screened by multiplex PCR assay for the stx1, stx2, eae and Ehly virulence genes. STECs were recovered from 8 per cent of diarrhoeic calves and 10.3 per cent of non-diarrhoeic calves. The predominant virulence gene profile was stx1/eae/Ehly (47.3 per cent) among isolates from diarrhoeic calves and eae/Ehly (36.8 per cent) among isolates from non-diarrhoeic calves. Among three tested serogroups, the predominant serogroup was O26 (18.4 per cent), and O157:H7 was not detected. Intimin subtyping by restriction fragment length polymorphism analysis revealed only three intimin subtypes (β, γ and θ). A significant difference was observed in the distribution of Int-θ between two groups. Int-6 was present in 50 per cent of the isolates from diarrhoeic calves and in 11.1 per cent of the isolates from non-diarrhoeic calves; this difference was statistically significant (P=0.01).
Magnesium Research (09531424)(2)
The aim of present study was to assess the effects of chronic administration of calcium-magnesium soft gels on the development of morphine tolerance and dependence in mice. Tolerance was assessed using the tail-pinch test and withdrawal signs of morphine were precipitated by injecting naloxone 2 h after the final morphine injection. CalMag capsules were given in final doses of 50/25, 25/12.5 and 12.5/6.25 mg/kg based on calcium/magnesium ratio. Similar doses of Ca and/or Mg were prepared, separately. CalMag at 50/25, 25/12.5 mg/kg and the mixture of calcium and magnesium (Ca + Mg) at 50/25, 25/12.5, 12.5/6.25 mg/kg and calcium at 50, 25 mg/kg significantly reduced the number of jumps. The number of standings was only reduced after the administration of CalMag at 50/25 mg/kg and Ca + Mg at 25/12.5 mg/kg. The development of morphine tolerance was prevented in all drug-treated groups, except the one which received 6.25 mg/kg Mg. The data suggested that combination of calcium and magnesium at 50/25 and 25/12.5 mg/kg prevented the development of tolerance and dependence. It seems that other ingredients of CalMag capsules do not have an important effect on preventing tolerance and withdrawal signs. Compared to the acute effects, chronic administration of CalMag allowed the effective dose to be reduced. Unlike the acute treatment, chronic administration of calcium alone was effective in reducing morphine tolerance and dependence, and magnesium had no significant effect on withdrawal signs, suggestive of some pharmacological adaptations.
Glucocorticoid hormones evidently affect memory. Morphine withdrawal causes a cognitive deficit and an increase in corticosterone secretion. In the present study brain corticosterone concentrations were determined after morphine withdrawal. Male mice were made dependent by increasing doses of morphine over 3 days. Blood and brain samples were collected following withdrawal induced by injection of naloxone (0.1 mg/kg) or spontaneously after 4 and 14 h. Brain corticosterone was extracted and measured by enzyme immunoassay. Short-term memory was determined in the novel object recognition task, using a 20 min interval between the learning trial and the test trial. In this memory trial, the difference in exploration between a previously seen object and a novel object is taken as an index of memory performance (recognition index, RI). RI in morphine dependent mice undergoing withdrawal was decreased compared to their control group. Brain corticosterone concentrations after naloxone withdrawal or 4 h after spontaneous withdrawal were respectively 22 and 34% greater than control values. Corticosterone concentration was normalized 14 h after the last dose of morphine. The results indicate that increase in brain corticosterone concentration may play an important role in short-term memory impairment following morphine withdrawal. © 2009 Informa UK Ltd All rights reserved.
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 (AT 1R) 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. Methods: 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 conclusion: 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.
Comparative Clinical Pathology (1618565X)(4)
Brucellosis is a major health problem worldwide especially in developing countries. It is an endemic disease in Iran. The transmission of brucellosis to man is primarily by direct contact with infected animals or their products. However, the organisms can also be transmitted by transfusion of infected blood. In this study, we determined serological prevalence of brucellosis among volunteer blood donors in Boushehr. A total of 10,500 blood donors were screened for serological evidence of infection with brucella spp. using the Rose Bengal plate test (RBPT). To confirm positive RBPT results, a standard tube agglutination test (STAT) and a 2-mercaptoethanol test (2ME) were performed. Of the 10,500 serum samples from blood donors in Boushehr province, Iran, we found anti-brucella antibodies in 0.076 and 0.057% sera samples with RBPT and STAT, respectively. The 2ME test was positive at low titer (1/20) in one sample only. © Springer-Verlag London Limited 2007.
Evidence-based Complementary and Alternative Medicine (17414288)(2)
The aim of the present study was to evaluate the anxiolytic effects of hydroalcoholic extract (HE) of Nepeta persica Boiss. (Lamiaceae) on the elevated plus-maze (EPM) model of anxiety. The extract of arial parts of the plant was administered intraperitoneally to male NMRI mice, at various doses, 30 min before behavioural evaluation. The HE extract of N. persica at the dose of 50 mg kg-1 significantly increased the percentage of time spent and percentage of arm entries in the open arms of the EPM. This dose of plant extract affected neither animal's locomotor activity nor ketamine-induced sleeping time. The 50 mg kg-1 dose of the plant extract seemed to be the optimal dose in producing the anxiolytic effects, lower or higher doses of the plant produce either sedative or stimulant effects. At 100 mg kg -1, the plant extract increased the locomotor activity. These results suggested that the extract of N. persica at dose of 50 mg kg-1 possess anxiolytic effect with less sedative and hypnotic effects than that of diazepam and causes a non-specific stimulation at 100 mg kg-1. © 2007 The Author(s).
Basic and Clinical Pharmacology and Toxicology (17427843)(4)
Morphine withdrawal leads to an increase in corticosterone concentration in plasma, and cognitive deficits are found after withdrawal. Evidence indicates that glucocorticoid hormones affect memory. The aim of the current study was to evaluate the effects of metyrapone and mifepristone on memory deficit following spontaneous morphine withdrawal. Memory was tested by using the object recognition task. The novel object recognition task was carried out in a square wooden open-field apparatus using objects. The test was comprised of three sections: habituation for 15 min., first trial for 12 min. and test trial for 5 min. In this learning paradigm, the difference in exploration between a previously seen object and a novel object is taken as an index of memory performance (recognition index - RI). Male mice were made dependent by increasing doses of morphine (30-90 mg/kg) subcutaneously twice daily for 3 days. RI was assessed 4 hr after the last dose of morphine on the third day. Mifepristone (50 and 100 mg/kg) and metyrapone (12.5 and 25 mg/kg) were used subcutaneously before the first trial and effects were compared to control values. Metyrapone (25 mg/kg) and mifepristone (50 mg/kg) improved RI to 34.8 ± 10.8% and 25.4 ± 11.7%, respectively, which are significantly different from control values (RI = -14.8 ± 10.7%, P < 0.05). These results show that increased glucocorticoid concentration may be involved in memory deficit caused by morphine withdrawal. Metyrapone by inhibiting glucocorticoid formation, and mifepristone by inhibiting glucocorticoid receptors may be useful for preventing memory deficit following morphine withdrawal. © 2008 The Authors.
Pakistan Journal of Biological Sciences (18125735)(23)
This study hypothesized that the expression of apoptosis-regulatory genes, such as BCL-2 and BAX may be affected by genetic variation in bleomycin-induced pulmonary fibrosis in C57BL/6 and NMRI mice. Pulmonary fibrosis induced by single intratracheal dose of bleomycin (3 U kg-1). After 2 weeks, lung samples were analyzed for collagen deposition, pathological changes and expression of BCL-2 and BAX. The fibrotic lung changes were similar in both strains. The immunohistochemical assay using a biotin-streptavidin technique showed no significant difference in immunoreactivity for BCL-2 protein between the controls and bleomycin-treated C57BL/6 mice. However, in NMRI mice, the expression of BCL-2 was significantly (p<0.05) upregulated in myofibroblasts and neutrophils. The expression of BAX protein was significantly (p<0.05) upregulated in alveolar epithelial cells of both strains and downregulated in myofibroblasts and lymphocytes of the lung tissues of C57BL/6 mice and also in lymphocytes of NMRI mice at 2 weeks after bleomycin instillation. These results confirm the role of BCL-2 and BAX proteins in the pathogenesis of pulmonary fibrosis and suggest that the expression of apoptotic regulatory genes may be specific in different cell types in various strains. © 2008 Asian Network for Scientific Information.
Pakistan Journal of Biological Sciences (10288880)(6)
The DNA sequence of Kringle-2 and serine protease domains of the human tissue plasminogen activator (reteplase, K2S) was PCR amplified. This product was then cloned into the expression vector pET15b plasmid. The presence of the insert was confirmed by restriction digestion, PCR and determination of the nucleotide sequence. By using isopropyl β-D thiogalactopyranoside (IPTG), reteplase was induced in E. coli BL21 cells and analyzed using polyacrylamide gel electrophoresis (PAGE). © 2007 Asian Network for Scientific Information.
Bleomycin-induced pulmonary fibrosis is a widely used experimental model for human lung fibrosis. The severity of fibrosis varies among different strains of mice and investigation on different strains and finding the mechanisms of variation is important in understanding the pathogenesis of human lung fibrosis. In the present study, NMRI mice were used to investigate the severity and also time-course of bleomycin-induced pulmonary fibrosis in comparison with C57BL/6 mice. After single dose administration of intratracheal bleomycin, the fibrotic response was studied by biochemical measurement of collagen deposition and semiquantitative analysis of pathological lung changes. NMRI mice developed lung fibrosis from 1 to 4 week after bleomycin instillation, with significant increases in lung collagen content and significant morphological changes (P < 0.05). These findings indicate that NMRI mice might be suitable as an experimental model of bleomycin-induced lung fibrosis.
Journal of Isfahan Medical School (10277595)(84)
Background: The p53 tumor suppressor gene plays an important role in genomic stability. A common G-to-C polymorphism at codon 72 in the p53 gene has been accompanied with high risk of lung, nasopharyngeal, oral, prostate, and colorectal cancers, and may result in genetic susceptibility to breast cancer. We studied the effect of this p53 polymorphism on breast invasive ductal carcinoma development Methods: This case-control study was conducted among 51 patients with breast invasive ductal carcinoma and 51 matched controls in Isfahan. P53 codon 72 genotypes were identified using allele-specific polymerase chain reaction (PCR). Findings: In control samples, the genotype distribution of p53 polymorphism showed 43.2%, 52.9% and 3.9% for the Arg/Arg, Arg/Pro and Pro/Pro genotypes, respectively. In cancer group the distribution was 86.2% for Arg/Arg, 11.8% for Arg/Pro and 2% for Pro/Pro. Distribution differences in p53 codon 72 polymorphism between the cases and controls were statistically significant (P<0.001). Conclusion: This study indicates that p53 codon 72 polymorphism is a genetic predisposing factor for breast invasive ductal carcinoma development in samples of Isfahan. However, further studies are needed to determine the role of p53 codon72 polymorphism in breast cancer development. © 2007, Isfahan University of Medical Sciences(IUMS). All rights reserved.
Journal of Camel Practice and Research (09716777)(1)
Pakistan Journal of Biological Sciences (18125735)(2)
The purpose of this study was to determine the presence of selected virulence genes in Escherichia coli isolated from diarrheic calves (Tehran province) and lambs (Ghume provinces) of Iran. We examined 40 isolates (29 from calves and 11 from lambs). All 40 isolates were tested for the presence of stx1, stx2, eae, espB and hly genes by multiplex polymerase chain reaction in two protocols. In the first protocol the isolates were tested with EC and hly primers and in the second protocol the isolates were examined with eae, stx1, stx2 and espB primers. Multiplex PCR showed that from 29 strains isolated from diarrheic calves, 4 of isolates (13.7%) were stx1 positive, 16 isolates (55.17%) carried stx2 and 2 isolates (6.89%) had espB gene. 1 isolate (3.44%) possessed eae. Among 11 strains isolated from diarrheic sheep, 9 isolates (81.81%) carried stx2 and 2 isolates (18.18%) had espB gene. Six isolates (54.54%) possessed eae and none of them was stx1 positive. The hly gene was not detected in any of the isolates. The findings of this study indicated that the stx2 may be widespread among pathogenic Escherichia coli in Iran. © 2006 Asian Network for Scientific Information.
Magnesium Research (09531424)(1)
The present study was aimed at evaluating the acute effects of Calcium-Magnesium soft gels (CalMag) in morphine tolerant and dependent mice. Mice were rendered tolerant and dependent on morphine by subcutaneous injection of morphine over a fixed time period. Withdrawal signs were precipitated by injecting naloxone 2 h after the final injection of morphine. The tail-pinch assay was used to investigate the effects of various compounds on the development and reversal of morphine tolerance. Acute injection of CalMag (containing 50 mg/kg calcium and 25 mg/kg magnesium) significantly reduced the number of jumps, stands and fast breathing in morphine dependent mice. Co-administration of calcium (50 mg/kg) and magnesium (25 mg/kg) was also effective in preventing the development of morphine tolerance and dependence. Administration of calcium (up to 50 mg/kg) alone did not significantly block the development of tolerance and dependence. The mean latency to pain was significantly increased in animals pretreated with CalMag (containing 50 mg/kg calcium and 25 mg/kg magnesium). The mixture of calcium and magnesium at specific concentrations seem to be critical for preventing the development of morphine tolerance and dependence.
Journal of Ethnopharmacology (03788741)(1-3)
The anxiolytic and sedative effects of hydroalcoholic extract (HE) of Salvia reuterana Boiss. was evaluated in mice. The HE of Salvia reuterana (100 mg/kg), increased the percentage of time-spent and the percentage of arm entries in the open arms of the elevated plus-maze. Spontaneous locomotor activity count measured in 15 min of the test was significantly decreased in animals pretreated with diazepam and 100 mg/kg of Salvia reuterana extract. Results of the present study provide support for the traditional usage of Salvia reuterana as a sedative and anxiolytic medicinal plant. © 2005 Elsevier Ireland Ltd. All rights reserved.
Phytotherapy Research (0951418X)(10)
The present study aimed to investigate the anxiolytic effects of four fractions of Stachys lavandulifolia Vahl. The aerial parts of the plant were extracted with petroleum ether (PF), ethyl acetate (EF), butanol (BF) and water (AF) and tested for spontaneous motor activity and elevated plus-maze (EPM) behaviour in mice. The hydroalcohol extract (HE) and different fractions of S. lavandulifolia were administered intraperitoneally to male Syrian mice, at various doses, 30 min before the behavioural evaluation. The HE of S. lavandulifolia (at 50 mg/kg) increased the percentage of time spent (39%) and the percentage of arm entries in the open arms (53%). The HE (50 mg/kg), PF (25 and 50 mg/kg), EF (25 and 50 mg/kg) and AF (50 mg/kg) of S. lavandulifolia significantly increased the percentage of time spent and the percentage of arm entries in the open arms. The BF up to a dose of 50 mg/kg had no significant effects on any of the measured parameters in the EPM. The spontaneous locomotor activity was significantly decreased in animals injected with each plant fractions, compared with that of saline. The EF and AF showed the least and the most reduction in the activity, respectively. The anxiolytic effects of EF, PF and AF could be related to their content of flavonoids, phenylpropanoids or terpenoids. Copyright © 2005 John Wiley & Sons, Ltd.
Biotechnology (1682296X)(4)
The present study was conducted to create mutations in this motif for further investigation of its function. By using nested PCR two mutations were produced, which would translate to DRY and DKH. The PCR products as well as the PGEM3Z vector were digested using NooI and EcoRI restriction enzymes and were ligated and transformed to E. coli HB101 cells. The obtained colonies were analyzed for the presence of the inserts using suitable restriction enzymes. The obtained plasmids have the advantage of having restriction sites, which would not interfere with further cloning and expression of this receptor using mammalian expression vectors. © 2005 Asian Network for Scientific Information.
Iranian Journal of Pharmaceutical Research (17350328)(2)
There is growing evidence indicating that neuronal calcium channels play an important role in the mechanism of morphine dependence. In this study, the effects of acute and chronic administration of nitrendipine on naloxone precipitated morphine withdrawal signs were investigated. Mice were rendered dependent to morphine by subcutaneous injection of morphine over a period of 5 days. In chronic studies, nitrendipine (25 and 50 mg/kg, i.p.), or vehicle injections were given once a day during the morphine treatment, and the last injection of nitrendipine was given 24 h before the morphine withdrawal. For acute studies, nitrendipine (25 and 50 mg/kg, i.p.) was given 1 h after the last dose of morphine (1 h before naloxone). A single injection of nitrendipine at 25 mg/kg was ineffective in blocking most signs of morphine withdrawal, however, at 50 mg/kg nitrendipine blocked signs such as hair raising, sniffing, diarrhea and number of jumping. The concurrent injections of nitrendipine with morphine prevented most signs of morphine withdrawal. In agreement with previous findings, these results suggest that alterations in voltage-sensitive calcium channels play a role in the adaptations that occur on chronic treatment with morphine. © 2004 by School of Pharmacy.
Fitoterapia (0367326X)(5)
The ethanolic extract of Echium amoenum flowers at the dose of 50 mg/kg increased the percentage of time-spent and the percentage of arm entries in the open arms of the elevated plus-maze (EPM) and decreased the percentage of time-spent in the closed arms of EPM. Moreover, it prolonged the ketamine-induced latency to sleep but had no significant effects on total sleeping time induced by ketamine. Also, the locomotor activity was affected but not to the same extent as observed for diazepam. These results suggested that the extract of E. amoenum seems to possess anxiolytic effect with lower sedative activity than that of diazepam. © 2004 Elsevier B.V. All rights reserved.
Iranian Journal of Pharmaceutical Research (17350328)(3)
Effect of Otostegia persica on naloxone-induced morphine withdrawal syndrome was studied in male mice. Dependence was induced using daily subcutaneous injections of morphine for three days. Morphine was injected to mice at doses of 30 and 45mg/kg on day 1 and 60 and 90mg/kg on day 2 (8:00 am and 6:00 pm). On day 3, morphine (90mg/kg) was injected 1h before oral administration and 1.5h before intraperitoneal (i.p.) injection of hydroalcoholic and hexane extracts of the plant. Naloxone was injected (5mg/kg, i.p.) 2h after the final dose of morphine and the withdrawal signs including jumping, rearing, diarrhoea, piloerection, tremor and ptosis were recorded during a period of 30 minutes. While oral and i.p. administration of hydroalcoholic extract reduced the number of jumping and rearing, the hexane extract could not exert any significant change. Also the hydroalcoholic extract (1500mg/kg) significantly (p<0.05) reduced diarrhoea, piloerection, tremor and ptosis. The hexane extract only significantly (p<0.05 ) inhibited diarrhoea. Results of this study indicated that the extract of Otostegia persica contained component(s) that alleviate morphine withdrawal syndrome and the responsible constituent(s) is(are) found in polar fraction since the hexane extract had only a negligible effect. © 2004 by School of Pharmacy Shaheed Beheshti University of Medical Sciences and Health Services.
Journal of Ethnopharmacology (03788741)(2-3)
Interest in alternative medicine and plant-derived medications that affect the "mind" is growing. The aim of the present study was to investigate the effects of a hydroalcoholic extract and essential oil of Stachys lavanduifolia Vahl on the elevated plus-maze (EPM) model of anxiety. The Stachys lavandulifolia extract or its essential oil was administered intraperitoneally to male TO mice, at various doses, 30min before the behavioral evaluation. The extract of Stachys lavandulifolia at the dose of 100mg/kg increased the percentage of time spent and the percentage of arm entries in the open arms of the EPM and decreased the percentage of time spent and the percentage of arm entries in the closed arms of the EPM. The plant extract at doses lower than 100mg/kg had no significant effects on any of the parameters measured on the EPM. This dose of the plant extract prolonged the ketamine-induced sleeping time, and decreased the locomotor activity in mice. These results suggested that the extract of Stachys lavandulifolia possessed anxiolytic effect with relatively lower sedative activity than diazepam. The essential oil of Stachys lavandulifolia, however, at doses of up to 100mg/kg did not have any significant effects on the mice behaviour on the EPM. © 2003 Elsevier Ireland Ltd. All rights reserved.
European Journal of Pharmacology (00142999)(1)
Characteristic changes of platelet membrane adenylyl cyclase activity have been described in men with alcoholism. We studied the occurrence of these changes in human erythroleukemia (HEL) cells after chronic ethanol treatment. Chronic treatment of the HEL cell with ethanol (50 or 100 mM) for 48 h resulted in significant reduction of prostaglandin E1-stimulated adenylyl cyclase activity. The acute ethanol (200 mM, 5 min) enhancement of adenylyl cyclase activity was significantly reduced after chronic ethanol treatment. We also observed a reduction in phorbol-12,13-dibutyrate (PDB) enhancement of prostaglandin E1-stimulation after chronic ethanol treatment. Chronic ethanol treatment (50 or 100 mM) reduced the activity of adenylyl cyclase in response to stimulation by acute ethanol to a greater extent than that of after acute PDB. The increase in cAMP formation by ethanol and PDB was only evident when prostaglandin E1 was present and under basal conditions (when no stimulatory agent was present) ethanol up to 200 mM, and PDB up to 1 M, had no significant effect on adenylyl cyclase activity. The reduced capacity of ethanol and/or PDB to stimulate adenylyl cyclase activity after chronic ethanol treatment suggests the involvement of a common denominator in the action of ethanol and PDB. © 2001 Elsevier Science B.V. All rights reserved.
Iranian Journal Of Medical Sciences (02530716)(3-4)
Background: The cAMP signal transduction systems are known to play a critical role in the acute and chronic effects of ethanol and other drugs of abuse. Objective: To investigate the role of protein kinase C (PKC) in the action of ethanol on platelets and human erythroleukemia (HEL) cells. Methods: HEL, HEK 293 cells [transfected with AC type VII(AC7)] and platelets were used to study the effects of ethanol. cAMP accumulation assay was used to determine the percentage conversion of [3H]ATP into [3H]cAMP. Each fraction was separated through sequential chromatography and the radioactive material was then quantified using a liquid scintillation counter. Results: Ethanol enhanced the PGE1-stimulated AC activity in HEL cells. The percent enhancement of cAMP accumulation by 200 mM ethanol was similar in HEL cells, platelets and HEK 293 cells transfected with AC7. When combined with PDBu, 200 mM ethanol further enhanced the PDBu stimulation of AC activity suggesting a separate mechanism by which ethanol and PDBu exert their effects on the AC. Pretreatment of the cells with staurosporine and chelerythrine significantly blocked the PDBu-and/or ethanol-enhancement of cAMP accumulation. Conclusion: These results indicate a clear role for PKC in the action of ethanol on AC in platelets.
European Journal of Pharmacology (00142999)(2-3)
Chronic barbital treatment significant increased the net K+-stimulated uptake of 45Ca2+ in cerebrocortical synaptosomal preparations, 24 h after withdrawal from chronic barbital administration. Basal uptake was not significantly changed. Hippocampal synaptosomal preparations showed a similar pattern, but the increase was not significant. The synaptosomal Ca2+ uptake was not affected by incubation with the dihydropyridine Ca2+ channel antagonist, nitrendipine, in controls or after chronic barbital treatment. Acute administration of a single dose of barbital did not alter the basal or stimulated uptake of 45Ca2+ in cortical synaptosomes, when this was measured 36 h after the barbital administration. Hippocampal slices prepared 24 h after withdrawal from chronic barbital treatment showed a significant increase in K+-stimulated uptake of 45Ca2+, and the basal uptake was significantly decreased. Both changes were prevented by nitrendipine. An increase in the density of dihydropyridine-sensitive binding sites was found in the cerebral cortex. The results indicate that both dihydropyridine-sensitive and insensitive neuronal Ca2+ channels are altered by chronic barbiturate treatment. These changes may be involved in physical dependence on barbiturates. Copyright (C) 1999 Elsevier Science B.V.
Alcoholism: Clinical and Experimental Research (01456008)(1)
Ethanol is known to enhance the activity of adenylyl cyclase (AC) in a number of cells and tissues. Recent work has suggested that the various isoforms of AC show differential sensitivity to ethanol, with Type VII AC being most sensitive. However, the mechanism of action of ethanol is unclear. In the present work, we investigated the effect of ethanol on AC activity in the human erythroleukemia (HEL) cell line, platelets, and AC VII-transfected HEK 293 cells. The HEL cells contain abundant amounts of mRNA for Type VII AC. We found that both ethanol and phorbol dibutyrate (PDBu) treatment enhanced agonist (prostaglandin E1; PGE1)-stimulated AC activity in HEL cells, as well as in platelets and HEK 293 cells transfected with AC VII. Inhibitors of protein kinase C (PKC) blocked the stimulatory effects of both ethanol and PDBu. However, the effects of ethanol and PDBu on AC activity were additive, suggesting that the mechanisms of action of ethanol and PDBu were not identical. Furthermore, a 30-min exposure of HEL cells to ethanol attenuated (desensitized) the ability of ethanol, but not PDBu, to enhance agonist-activated AC activity. On the other hand, a 30-min pretreatment with PDBu attenuated the AC response to the phorbol ester, but not to ethanol; but, after a 20 hr preincubation with phorbol ester, the ability of both PDBu and ethanol to enhance prostaglandin E1-stimulated AC activity was completely eliminated. Finally, pretreatment of HEL cells with pertussis toxin blocked the effect of PDBu, but not ethanol, on AC activity. The results support the involvement of phorbol ester-sensitive PKC(s) in ethanol's enhancement of agonist-activated activity of AC in HEL cells, but suggest that the mechanism of ethanol's action is different from that of PDBu. The findings with pertussis toxin suggest that PDBu activation of PKC(s) may affect AC activity through phosphorylation of a G(i) protein, whereas ethanol may act by promoting phosphorylation of a different substrate (e.g., AC VII).
Pharmacology, Biochemistry and Behavior (00913057)(1)
Tolerance occurred to the sedative actions of the competitive NMDA antagonists, CGP39551 and CGP37849, as measured by a decrease in spontaneous locomotor activity after 1 week or 2 weeks of administration, respectively, in studies using the TO strain of mice. Crosstolerance was seen between these compounds. When CGP37849 was given after 2 weeks treatment with CGP39551, an increase in locomotor activity was seen. Chronic barbiturate treatment, producing tolerance to the actions of pentobarbitone, did not affect the sedative properties of CGP39551 or CGP37849. Chronic treatment with CGP39551 did not alter the ataxic actions of pentobarbitone. Seven days of treatment with HA966 caused complete tolerance to its sedative actions, but no Crosstolerance was seen to pentobarbitone, CGP39551, or CGP37849. A small but significant decrease was seen in the convulsion thresholds to NMDA after 15 days of treatment with CGP39551, and a small significant increase in ratings of convulsive behavior after 16 days injections of CGP37849. No significant changes were found in either Bmax or Kd for [3H]-MK-801 binding in cerebrocortical tissue 24 h after the last chronic treatment with either of the NMDA antagonists. © 1994.
Pharmacology, Biochemistry and Behavior (00913057)(3)
We have shown previously that the dihydropyridine calcium channel antagonist nitrendipine, given chronically, prevents the development of ethanol tolerance and physical dependence. The present study examines the effects on barbiturate tolerance and physical dependence. Nitrendipine, given acutely during withdrawal, provided little protection against barbiturate withdrawal, as measured by convulsive behaviour on handling. When nitrendipine was given chronically concurrently with the barbiturate, a prolonged protection against the withdrawal syndrome was seen. Acute nitrendipine significantly increased the latency of seizures in response to the partial benzodiazepine inverse agonist FG7142 during barbiturate withdrawal, but there was no effect on the seizure incidence in response to bicuculline. Chronic treatment with nitrendipine did not alter the development of tolerance to the ataxic or general anaesthetic actions of barbiturates, but evidence was found of a possible interaction between nitrendipine and pentobarbitone, which may have been pharmacokinetic. The results suggest that neuronal calcium channels may be involved to some degree in the development of the changes responsible for barbiturate withdrawal, but to a less extent than found previously for ethanol dependence. © 1994.
British Journal of Pharmacology (00071188)(1)
The competitive antagonists at the N‐methyl‐d‐aspartate (NMDA) receptor, CGP39551 and CGP37849, protected against the barbiturate withdrawal syndrome in mice, as measured by ratings of convulsive behaviour on handling. The effective doses of these compounds were lower than those required to prevent seizures due to NMDA in naive animals; these were in turn lower than those needed to prevent the convulsive effects of the α‐aminobutyric acid (GABA) antagonist, bicuculline. The NMDA‐receptor antagonists did not alter the increase in the incidence of convulsions due to the GABAA antagonist, bicuculline, that is seen during barbiturate withdrawal, although the latencies to these convulsions during barbital withdrawal were significantly increased after CGP39551. Barbiturate withdrawal did not affect the convulsive actions of NMDA, whether measured by the incidence of convulsions or by intravenous infusion. The Bmax for [3H]‐dizocilpine ([3H]‐MK801) binding was significantly increased by chronic barbital treatment in cerebrocortical but not in hippocampal tissues, while the Kd remained unaltered in either case. At 1 h and 24 h after administration of a single dose of barbitone, the Bmax for [3H]‐dizocilpine binding was unaltered in cerebrocortical tissue. Acute addition of barbitone in vitro did not alter [3H]‐dizocilpine binding or the displacement of binding of thienylcyclohexylpyridine. 1994 British Pharmacological Society
Neuropharmacology (00283908)(3)
The effects of the dihydropyridine calcium antagonist, nitrendipine and the calcium channel activator, Bay K 8644, have been compared on the anaesthetic, ataxic and anticonvulsant effects of benzodiazepines. Possible interactions between the peripheral benzodiazepine receptor antagonist, PK11195, and the classical benzodiazepines were also examined. Nitrendipine considerably potentiated the anaesthetic effects of benzodiazepines and increased their ataxic effects but had no effect on the anticonvulsant actions. Clonazepam did not produce anaesthesia, at doses up to 1 g kg-1 or when given with nitrendipine. When given alone, nitrendipine did not cause general anaesthesia. Nitrendipine did not appear to alter the metabolism of midazolam. The calcium channel activator, Bay K 8644, reduced the anaesthetic potency of midazolam and, when given alone, produced ataxia. It did not significantly alter central concentrations of midazolam. The "peripheral" benzodiazepine antagonist, PK11195, did not affect the ataxic or anaesthetic actions of benzodiazepines. These results suggest that dihydropyridine-sensitive calcium channels may be more important to the general anaesthetic than to the anticonvlsant actions of benzodiazepines. The "peripheral" benzodiazepine site did not appear to play a role in either of these properties. © 1991.
British Journal of Pharmacology (00071188)(3)
1. The effects of the calcium channel blocking agent, nitrendipine, were studied on seizures in mice produced during withdrawal from chronic benzodiazepine treatment and on the development of tolerance to benzodiazepines. 2. Nitrendipine produced a dose-dependent decrease in seizure incidence, when seizures were produced by the partial inverse agonist FG7142 during withdrawal from seven days treatment with flurazepam. 3. Nitrendipine did not raise the seizure thresholds in naive mice to the full inverse agonist methyl-6,7-dimethoxy-4-ethyl-β-carboline-3-carboxylate (DMCM), or to the γ-aminobutyric acid (GABA) antagonist, bicuculline. 4. When given concurrently with flurazepam for seven days, nitrendipine did not affect the incidence of seizures during flurazepam withdrawal. 5. When given concurrently with the benzodiazepines, nitrendipine did not prevent the development of tolerance to midazolam general anaesthesia or tolerance to the ataxic actions of flurazepam or midazolam. 6. Chronic treatment with flurazepam for seven days did not affect the K(d) or B(max) of [3H]-nimodipine binding in mouse whole brain or cerebral cortex. 7. These results with benzodiazepines are partially in contrast with those for ethanol, where nitrendipine not only decreased ethanol withdrawal seizures when given acutely, but also prevented the development of tolerance and withdrawal signs when given concurrently with ethanol. However, they do confirm the selectivity of nitrendipine for withdrawal-induced seizures.
