Advanced Pharmaceutical Bulletin
Volume:3 Issue: 2, Dec 2013

Gene therapy is one of the most attractive fields in medicine. The concept of gene delivery to tissues for clinical applications has been discussed around half a century, but scientist’s ability to manipulate genetic material via recombinant DNA technology made this purpose to reality. Various approaches, such as viral and non-viral vectors and physical methods, have been developed to make gene delivery safer and more efficient. While gene therapy initially conceived as a way to treat life-threatening disorders (inborn errors, cancers) refractory to conventional treatment, to date gene therapy is considered for many non–life-threatening conditions including those adversely influence on a patient’s quality of life. Gene therapy has made significant progress, including tangible success, although much slower than was initially predicted. Although, gene therapies still at a fairly primitive stage, it is firmly science based. There is justifiable hope that with enhanced pathobiological understanding and biotechnological improvements, gene therapy will be a standard part of clinical practice within 20 years.

The production of proteins in appropriate quantity and quality is an essential requirement of the present time. There appears to be a progressive increase in the application of mammalian cells for proteins production. Expression systems utilizing mammalian cells for recombinant proteins are able to introduce proper protein folding, post-translational modifications, and product assembly, which are important for complete biological activity. This review article is totally based on literature survey. In this article much emphasis has been done on the mammalian expression system. The author focused on different mammalian cell lines that express the gene. The different vector systems that transfer the gene into mammalian cells like plasmid based expression vectors, adenovirus vectors, vaccinia vectors, retroviral vector and baculovirus as vectors were explored. The processes for the transfer of gene into mammalian cells were also reviewed. Application and limitations of mammalian expression system were also focused. The purpose of research in writing this article is to create awareness in researchers, starting their career in gene expression related to mammalian cells. The principal result and major conclusion of this article is to make available the molecular technologies, expression system and applications of gene expression in mammalian cell lines.

Ghrelin has been shown to have antiepileptic function. However, the underlying mechanisms by which, ghrelin exerts its antiepileptic effects are still unclear. In the present study, we investigated whether neuropeptide Y (NPY) mediates ghrelin anticonvulsant effect in the brain through its Y1, Y2 or Y5 receptors. Male Wistar rats were bilaterally microinjected with ghrelin 0.3 nmol/μl/side and NPY antagonists; GR231118 (Y1 receptor antagonist), BIIE0246 (Y2 receptor antagonist), CGP71683 (Y5 receptor antagonist) or solvents (Saline, DMSO) into the dorsal hippocampus 20 minutes before ghrelin administration. Thirty minutes after ghrelin microinjection, a single convulsive dose of pentylenetetrazole (PTZ) (50 mg/kg) was injected intraperitoneally (ip). Afterwards, duration of seizure and total seizure score (TSS) were assessed for 30 minutes in all animals. Intrahippocampal injection of 0.3 nmol/μl/side ghrelin decreased duration of seizure and TSS induced by PTZ. The suppression of both duration (p< 0.001) and TSS (p< 0.001) induced by ghrelin in hippocampus were significantly blocked by GR231118 (10 μg/μl/side), BIIE0246 (400 pmol/μl/side) and CGP 71683A (5 nmol/μl/side). Our findings suggest that NPY Y1, Y2 and Y5 receptors in the hippocampus may somehow mediate the anticonvulsive action of ghrelin. Therefore, it is possible to speculate that ghrelin acts in the hippocampus to modulate seizures via NPY.

Low-grade inflammation, a common feature in type 2 diabetes (DM2), causes some chronic complications in these patients. The present study was aimed to evaluate the effects of ginger (Zingiber officinale) on pro-inflammatory cytokines (IL-6 and TNF-α) and the acute phase protein hs-CRP in DM2 patients as a randomized double-blind placebo controlled trial. A total of 64 DM2 patients randomly were assigned to ginger or placebo groups and received 2 tablets/day of each for 2 months. The concentrations of IL-6, TNF-α and hs-CRP in blood samples were analyzed before and after the intervention. Ginger supplementation significantly reduced the levels of TNF-α (P = 0.006), IL-6 (P = 0.02) and hs-CRP (P = 0.012) in ginger group in comparison to baseline. Moreover, the analysis of covariance showed that the group received ginger supplementation significantly lowered TNF- α (15.3 ± 4.6 vs. 19.6 ± 5.2; P = 0.005) and hs-CRP (2.42 ± 1.7 vs. 2.56 ± 2.18; P =. 016) concentrations in comparison to control group. While there were no significant changes in IL-6 (7.9 ± 2.1 vs. 7.8 ± 2.9; P >. 05). In conclusion, ginger supplementation in oral administration reduced inflammation in type 2 diabetic patients. So it may be a good remedy to diminish the risk of some chronic complications of diabetes.

Carpal tunnel syndrome (CTS) refers to a cluster of signs and symptoms that stems from compression of the median nerve traveling through carpal tunnel. Surgery is a definite treatment for CTS; however, many conservative therapies have been proposed. The present study set out to assess the effect of vitamin B6 in patients with CTS. Forty patients (67 hands) with mild-moderate CTS were initially selected and randomly assigned into two groups as follows: 1) Case group with 20 subjects (32 affected hands) receiving vitamin B6 (120 mg/day for 3 months) and splinting. 2) Control group with 19 subjects (35 affected hands) only received splinting. One subject from the control group dispensed with continuing participation in the research. Daily symptoms and electrodiagnostic (NCV-EMG) results were assessed at baseline and after 3 months. Nocturnal awakening frequency due to pain, daily pain, daily pain frequency, daily pain persistence, hand numbness, hand weakness, hand tingling, severity of nocturnal numbness and tingling, nocturnal awakening frequency owing to hand numbness and tingling, and clumsiness in handling objects improved significantly in the vitamin B6-treated patients; even so, only problem with opening a jam bottle and handling phone significantly reduced in the control group. The median nerve sensory latency mean decreased following the treatment; and the median nerve sensory amplitude mean and sensory conduction velocity mean increased. The present study suggests that vitamin B6 treatment improves clinical symptoms and sensory electrodiagnostic results in CTS patients, and thus is recommended for CTS treatment.

Standardization and detailed pharmacognostical studies of Oreganum vulgare Linn. leaf for authentication and commercial utilization. Oreganum vulgare Linn. leaf was with standardization according to standard procedures described in WHO, 2011 and I.P. 1996. The physicochemical parameters total ash, acid insoluble ash, water soluble ash and sulphated ash were found to be 11.5%, 11%, 5, 10.5% w/w respectively. Foaming index was found be <100. The trace elements were found to be copper, lead, cadmium, zinc, cobalt, manganese, nickel and copper in ethanol extract and phytochemical screening of aqueous and ethanol extract showed the presence of carbohydrates, flavonoids, anthocyanins, phenolic compounds etc. The standardization parameters viz. physico-chemical parameters, macroscopy, microscopy, taxonomy, anatomy and preliminary phytochemical screening, microbial and aflatoxin count, HPTLC profile is being reported to help in authentication and development of monograph of this plant.

The aim of the present study was to develop a simple and rapid reversed-phase high performance liquid chromatographic method with UV detection for erlotinib hydrochloride quantification, which is applicable for protein binding studies. Ultrafilteration method was used for protein binding study of erlotinib hydrochloride. For sample analysis a simple and rapid reversed-phase high performance liquid chromatographic method with UV detection at 332 nm was developed. The mobile phase was a mixture of methanol, acetonitril and potassium dihydrogen phosphate buffer (15:45:40 %v/v) set at flow rate of 1.3 ml/min. The run time for erlotinib hydrochloride was approximately 6 minutes. The calibration curve was linear over the range of 320-20000 ng/ml with acceptable intra- and inter-day precision and accuracy. The intra-day and inter-day precisions were less than 10% and the accuracies of intra and inter-day assays were within the range of 97.20-104.83% and 98.8-102.2% respectively. Based on the obtained results, a simple, accurate and precise reversed-phase isocratic HPLC method with UV detection has been optimized and validated for the determination of erlotinib hydrochloride in biological samples.

This study was conducted to assess the effect of skin pre-treatment with Transcutol® and eucalyptus oil on systemic absorption of topical trolamine salicylate in rat. Pharmacokinetic parameters of salicylic acid following administration of trolamine salicylate on rat skin pre-treated with either Transcutol® or eucalyptus oil were determined using both non-compartmental and non-linear mixed effect modeling approaches and compared with those of control group. Median (% of interquartile range/median) of salicylic acid AUC0-8hr (ng/mL/hr) values in Transcutol® or eucalyptus oil treated rats were 2522(139%) and 58976(141%), respectively as compared to the 3023(327%) of the control group. Skin pre-treatment with eucalyptus oil could significantly decrease extravascular volume of distribution (V/F) and elimination rate constant (k) of salicylic acid. Unlike Transcutol®, eucalyptus oil lead to enhanced transdermal absorption of trolamine salicylate through rat skin.

Various cytokine regulates hematopoesis; they promote number of stages in stem cells biology such as proliferation, differentiation and endurance. Biological effects of SCF, as a hematopoietic cytokine; is triggered by binding to its ligand c-kit. Potential therapeutic applications of SCF include hematopoietic stem cell mobilization, exvivo stem/progenitor cell expansion, gene therapy, and immunotherapy. In this study we tried to construct of pPIC9 recombinant vector containing human SCF. hSCF cDNA was amplified by PCR and both hSCF cDNA and pPIC9 as yeast expression vector (shuttle vector) digested by EcoR I and Xho I restriction enzymes. Subsequent the digestion reaction, ligation reaction was carried out. In order to verifying of pPIC9 recombinant vector containing hSCF, PCR and sequence analysis was performed. The construction of recombinant expression vector of pPIC9 containing hSCF cDNA was confirmed by sequencing method successfully. rhSCF/pPIC9 vector can be transformed into the Picha pastoris yeast as a eukaryotic host in order to produce human SCF at industrial scale.

Hepatoprotective potentials of Jussiaea nervosa leaf extract against Cadmium-induced hepatotoxicity were investigated. Forty albino rats were randomly assigned into groups A-G with 4 rats in each of the groups A-F. Group A served as control and were given feed only while rats in groups B-F were orally exposed to varying concentrations of cadmium for six weeks. Effects of cadmium were most significant at 12 mg/Kg body weight (BW), and this dose was used for subsequent test involving oral administration of Jussiaea nervosa leaf extracts. In this segment, group G (n= 16) was sub-divided into four: G1-G4, with each sub-group containing four rats. Rats in sub-group G1 were given cadmium and feed only and served as positive control. Rats in sub-groups G2, G3, and G4 were given cadmium and 20, 50 and 100g/kg BW of Jussiaea nervosa extract, respectively, for six weeks. Blood and liver were analysed using standard laboratory techniques and methods. Liver function parameters (ALT, AST, ALP, bilirubin) were significantly (p<0.05) elevated in exposed rats in comparison to the controls, except for total protein and albumin, which were significantly decreased. Histopathological assessment reveals renal pathology in exposed rats in sharp contrast with the controls. Jussiaea nervosa extract however lowered the values of liver function parameters with 100mg/Kg BW dose producing the highest ameliorative effects. Similarly, the serum albumin and total protein significantly (p<0.05) improved with normal liver architecture. The results show the hepatoprotective potentials of Jussiaea nervosa extract against Cd toxicity.

The aim of this study was to design, formulate and physicochemically evaluate effervescent ranitidine hydrochloride (HCl) tablets since they are easily administered while the elderly and children sometimes have difficulties in swallowing oral dosage forms. Effervescent ranitidine HCl tablets were prepared in a dosage of 300 mg by fusion and direct compression methods. The powder blend and granule mixture were evaluated for various pre-compression characteristics, such as angle of repose, compressibility index, mean particle size and Hausner's ratio. The tablets were evaluated for post-compression features including weight variation, hardness, friability, drug content, dissolution time, carbon dioxide content, effervescence time, pH, content uniformity and water content. Effervescent systems with appropriate pre and post-compression qualities dissolved rapidly in water were selected as the best formulations. The results showed that the flowability of fusion method is more than that of direct compression and the F5 and F6 formulations of 300 mg tablets were selected as the best formulations because of their physicochemical characteristics. In this study, citric acid, sodium bicarbonate and sweeteners (including mannitol, sucrose and aspartame) were selected. Aspartame, mint and orange flavors were more effective for masking the bitter taste of ranitidine. The fusion method is the best alternative in terms of physicochemical and physical properties.

Anti cancer drugs is one of the most important chemotherapeutic factors whichcan influence spermatogenesis process and germinal epithelium. Since dividing cells aremainly affected by anticancer drugs, the aim of the present study is to investigate thepreventive effect of GnRH antagonist on spermatogenic defect produced by anticancerdrugs. In the present study thirty adult male mice aging 6-8 weeks were divided into 3 groups as: Control, Experimental 1 and Experimental 2. Experimental 1 group received Cisplatin for 5 days as 2.5 mg/kg intraperitoneally and Experimental 2 group received 0.25 mg/kg cetrorelix (GnRH antagonist) one week before cisplatin treatment and continued for 3 weeks. The mice in all groups were sacrificed 35 days after the last injection and testis specimens were fixed in boueins, formaldehyde fixative and 2.5% Glutaraldehide then prepared for light and electron microscopic examination. Light microscopy (LM) study showed that the number of spermatogonial cells, thickness of germinal epithelium, was decreased in Experimental 1group. Electron microscopy revealed that in this group several intercellular spaces appeared between spermatogenic cells and secretory granules in interstitial cells was increased. There were several vacuolated mitochondria and destroyed organelles in spermatogonial cells but in Experimental 2 group condition was similar to control group. These results indicate that the cetrorelix administration before cancer treatment may protect germinal epithelium against side effects of cisplatin.

CD14, one of the main differentiation markers on the surface of myeloid lineage cells, acts as a key role in activation of LPS-induced monocytes. LPS (lipopolysaccharide) binds to LPS-binding protein in plasma and are delivered to the cell surface receptor CD14. In this study, Various stimuli [Dimethyl Sulfoxide (DMSO), active 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] and LPS], either alone or in combination, have been recognized that have an effect on the level of CD14 expression in the human HL-60 and U937 promonocytic cell lines and therefore induce their terminal differentiation into monocytes or mature macrophages. U937 and HL-60 cells were cultured in RPMI 1640 supplemented with 10% FBS. For each cell line, 1×106 cells were seeded for 72 hours with DMSO, 14 days with LPS and 18 days with 1, 25-D3 in each well plate; then ELISA method was used to study their responses to the factors by means of anti-CD14. ELISA assay demonstrated that U937 and HL-60 cells were induced by both [1,25(OH)2D3] and DMSO to obtain characteristics of adherent cells and express CD14 protein; moreover, LPS at a low dose increased CD14 expression on surface of this cells. According to the our results, it is speculated that CD14 gene expression may be induced in human U937 and HL-60 cell lines by different factors including 1,25-D3, DMSO and LPS.

Monoclonal antibodies and related conjugates are key reagents used in biomedical researches as well as, in treatment, purification and diagnosis of infectious and non- infectious diseases. Balb/c mice were immunized with purified human IgG. Spleen cells of the most immune mouse were fused with SP2/0 in the presence of Poly Ethylene Glycol (PEG). Supernatant of hybridoma cells was screened for detection of antibody by ELISA. Then, the sample was assessed for cross-reactivity with IgM & IgA by ELISA and confirmed by immunoblotting. The subclasses of the selected mAbs were determined. The best clone was injected intraperitoneally to some pristane-injected mice. Anti-IgG mAb was purified from the animal's ascitic fluid by Ion exchange chromatography and then, mAb was conjugated with HRP. In the present study, over than 50 clones were obtained that 1 clone had optical density over than 3. We named this clone as supermonoclone which was selected for limiting dilution. The result of the immunoblotting, showed sharp band in IgG position and did not show any band in IgM&IgA position. Based on the findings of this study, the conjugated monoclonal antibody could have application in diagnosis of infectious diseases like Toxoplasmosis, Rubella and IgG class of other infectious and non- infectious diseases.

Cyclic voltammetry and differential pulse voltammetry were used to explore the diffusion behavior of two antipsychotic drugs at a glassy carbon electrode. A well-defined oxidation peak was obtained in Britton-Robinson (BR) buffer (pH 2.0). The response was evaluated as a function of some variables such as the scan rate, and pH. A simple, precise, inexpensive and sensitive voltammetric method has been developed for the determination of the cited drugs Olanzapine (OLZ) and Quetiapine fumarate (QUT). A linear calibration was obtained from 3 × 10-8 M to 4 × 10-6 M and 2 × 10-8 M to 5 × 10-6 M, with R. S. D. were 1.6 % and 1.2 % for OLZ and QUT, respectively. The limit of detection (LOD) was 1 × 10-8 M, while the limit of quantification (LOQ) was 3 × 10-8 M. The method was applied to the determination of investigated drugs in urine and serum samples and dosage forms.

The purpose of this study was to determine the effects of Phaleria macrocarpa (PM) on male fertility by assessing its effect on the sperm characteristics which included the sperm count, motility, viability and morphology. Eighteen male rats were equally divided into three groups. Each group of rats was orally supplemented for 7 weeks either with PM aqueous extract (240 mg/kg), distilled water (0 mg/kg) or testosterone hormone, Andriol® Testocaps™ (4 mg/kg) respectively. On the last day of supplementation period, the rats were sacrificed and sperm was obtained from cauda epididymis via orchidectomy. The sperm count, motility, viability and morphology were determined. PM aqueous extract significantly increased (p<0.05) the percentage of sperm viability. However, there was no significant effect of PM on the percentage of both sperm motility and morphology. The mean of body weight declined significantly in rats supplemented with PM aqueous extract compared to control groups (p<0.05). The results showed that PM significantly increased sperm viability without changing the sperm motility and morphology. Hence, this study suggests that PM offers an alternative way to improve male fertility by improving the sperm quality.

Ghrelin has been shown to have antiepileptic function. However, the underlying mechanisms by which, ghrelin exerts its antiepileptic effects are still unclear. In the present study; we investigated antiepileptic mechanism of ghrelin through GABAB receptors using CGP35348 (selective GABAB receptor antagonist). Male Wistar rat's hippocampi were bilaterally microinjected with the single dose or 10-day ghrelin (0.3 nmol/μl/side). CGP35348, GABAB receptor antagonist, (12.5 μg/μl/side) or saline injected into the dorsal hippocampus 20 minutes before ghrelin administration. Thirty min after ghrelin microinjection, a single convulsive dose of pentylenetetrazole (PTZ) (50 mg/kg) was injected intraperitoneally (i.p). Afterwards, seizure duration and total seizure score (TSS) were assessed for 30 minutes in all animals. Our results demonstrated that acute and chronic intrahippocampal (i.h.) injection of ghrelin could significantly (p<0.001) attenuate the severity of seizures. Ghrelin 0.3 nmol/μl/side decreased duration of seizure significantly both in acute (p<0.001) and chronic (p<0.01) injections. The ghrelin antiepileptic effect was completely antagonized by GABAB blockade. The suppression of both duration and TSS induced by ghrelin in hippocampus was significantly (p<0.001) blocked by CGP35348 in PTZ-induced seizures. In summary, our findings suggest that GABAB receptors may mediate the antiepileptic action of ghrelin in the hippocampus. Therefore, it is possible to speculate that ghrelin acts in the hippocampus to modulate seizures via GABA.

This study was made to investigate the antihyperglycemic and antioxidant potential of oil of seeds of Brassica nigra (BNO) in streptozotocin -nicotinamide (STZ) induced type 2 diabetic rats. BNO was orally administered to diabetic rats to study its effect in both acute and chronic antihyperglycemic study. The body weight, oral glucose tolerance test and biochemical parameters viz. glucose level, insulin level, liver glycogen content, glycosylated hemoglobin and antioxidant parameters were estimated for all treated groups and compared against diabetic control group. Administration of BNO at a dose 500 mg/kg and 1000 mg/kg body weight p.o. to STZ diabetic rats showed reduction in blood glucose level from 335 mg/dl to 280 mg/dl at 4th h and from 330 mg/dl to 265 mg/dl respectively which was found significant (p<0.01) as compared with diabetic control. BNO (500 mg/kg and 1000 mg/kg) and glibenclamide (0.6 mg/kg) in respective groups of diabetic animals administered for 28 days reduced the blood glucose level in streptozotocin-nicotinamide induced diabetic rats. There was significant increase in body weight, liver glycogen content, plasma insulin level and decrease in glycosylated hemoglobin in test groups as compared to control group. In vivo antioxidant studies on STZ-nicotinamide induced diabetic rat’s revealed decreased malondialdehyde (MDA) and increased reduced glutathione (GSH). Thus the results showed that the oil of seeds of Brassica nigra has significant antihyperglycemic and antioxidant activity.

The paper describes some thin layer chromatographic procedures that allow simple and rapid separation and identification of penicillins and cephalosporins from complex mixtures. Using silicagel GF254 as stationary phase and selecting different mobile phases we succeeded in the separation of the studied beta-lactamins. Our aim was not only to develop a simple, rapid and efficient method for their separation but also the optimization of the analytical conditions. No system will separate all the beta-lactams, but they could be identified when supplementary information is used from color reactions and/or by using additional chromatographic systems. The right combination of solvent system and detection method allows the identification of the studied penicillins and cephalosporins and can be successfully used in the preliminary analysis beta-lactam antibiotics.

To investigate postoperative pain control and analgesic use after heart surgery. 120 patients undergone heart surgery, randomly entered the study. Each patient was asked to score his pain intensity on visual analog scale (VAS) at four different occasions. 120 patients aged 59 year-old; including 81 male were enrolled in the study. 69.2% had coronary artery disease and 16.7% had heart-valve problem. Main types of surgeries were coronary artery bypass surgery (70.5%) and valve repairement (23%). Duration of ICU stay was 4.78±2.7 days and duration of intubations was 17.38 ± 36.46 hours. Pre-surgery pain relief was administrated to 42% of the subjects and morphine and promethazine was the main pre-surgery analgesia medication. Post surgery analgesic included morphine (injection), petidine (injection) and NSAIDS (oral or rectal). According to VAS, mean pain level, 1 and 4 hours after extubation, and before and one hour after transferring to wards was 5.05±2.5, 4.09±2.0, 3.52±1.8, 2.36±1.89, respectively. Although the level of pain reported was mostly moderate, 80% were reported satisfaction with their post-surgery pain management. A closer pain management control is needed for patients after heart surgery. Introduction of newer pain management techniques, medications and dosages could reduce the pain and suffering.

To evaluate phytochemical composition, antibacterial and antioxidant properties of methanolic extracts of different parts viz., leaves, follicles and latex of Indian devil tree (Alstonia scholaris Linn.) R. Br. Antibacterial activities of the methanol extracts against Gram +ve (Bacillus subtilis and Staphylococcus aureus) and Gram -ve (Escherichia coli, Pseudomonas aeruginosa) bacteria were determined by well diffusion techniques. Aantioxidant profiles of methanol extracts were determined by 1,1-diphenyl-2-picryl-hydrazil (DPPH) free radical scavenging, superoxide anion radial scavenging and ferric thiocyanate reducing assays. Phytochemical composition revealed abundance of flavonoids (97.3 mg QE/g DW), proanthocynidins (99.3 mg CE/g DW) and phenolics (49.7 mgGAE/g DW) in the leaf extract. Extracts of follicles and latex had comparatively very content of phenolics, flavonoids and proanthocyanidins. However, in follicle extract level of proanthocyanidins was significantly higher (46.8 mg CE/gDW). Latex extract among others exhibited most potent antibacterial activity. All the extracts displayed strong DPPH free radical and superoxide anion scavenging activities, only leaf extract displayed powerful reducing and ferrous ion chelating activities. Study revealed significant antioxidant activities of A. scholaris leaf, follicles and latex extracts and potential antibacterial activity of latex extract.

The purpose of this study was the isolation and structure elucidation of chemical compounds from the rhizomes of Eremostachys laciniata (L) Bunge (EL), an Iranian traditional medicinal herb with a thick root and pale purple or white flowers as well as the clinical studies on the therapeutic efficacy and safety of topical application of the EL extract in the management of some inflammatory conditions, e.g., arthritis, rheumatoid arthritis and septic arthritis (Riter’s syndrome). The structures of the isolated compounds were elucidated unequivocally on the basis of one and two dimensional NMR, UV and HR-FABMS spectroscopic data analyses. A single-blinded randomized clinical trial was carried out with the extract of the rhizomes of E. laciniata (EL) to determine the efficacy and safety of the traditional uses of EL compared to that of piroxicam in treatment of inflammatory diseases, e.g., osteoarthritis, rheumatoid arthritis and Reiter’s syndrome. Eleven iridoid glycosides, two phenylethanoids and two phytosterols were isolated and identified for the first time from the rhizomes of EL. After 14 days of treatment with the EL and piroxicam ointments, all groups showed significant improvements compared to the control groups. EL (5%) ointment induced better initial therapeutic response than piroxicam (5%) onitment. This clinical trial established that EL was suitable for topical applications as a safe and effective complementary therapy for inflammatory diseases.

Oxidative stress is generated through imbalance between composing and decomposing of reactive oxygen species (ROS). This kind of stress was rarely discussed in connection with foreign protein production in Escherichia coli. Effect of cytoplasmic recombinant protein expression on Hydrogen peroxide concentration and catalase activity was previously reported. In comparison with cytoplasm, periplasmic space has different oxidative environment. Therefore, in present study we describe the effect of periplasmic expression of recombinant human interleukin-2 (hIL-2) on H2O2 concentration and catalase activity in Escherichia coli and their correlation with cell growth. Having constructed pET2hIL2 vector, periplasmic expression of hIL-2 was confirmed. Then, H2O2 concentration and catalase activity were determined at various ODs. Wild type and empty vector transformed cells were used as negative controls. It was shown that H2O2 concentration in hIL-2 expressing cells was significantly higher than its concentration in wild type and empty vector transformed cells. Catalase activity and growth rate reduced significantly in hIL-2 expressing cells compared to empty vector transformed and wild type cells. Variation of H2O2 concentration and catalase activity is intensive in periplasmic hIL-2 expressing cells than empty vector containing cells. Correlation between H2O2 concentration elevation and catalase activity reduction with cell growth depletion are also demonstrated. Periplasmic expression of recombinant hIL-2 elevates the host cell’s hydrogen peroxide concentration possibly due to reduced catalase activity which has consequent suppressive effect on growth rate.

Analysis of drug utilized the organic solvent which are costlier, toxic and causing environment pollution. Hydrotropic solution may be a proper choice to preclude the use of organic solvents so that a simple, accurate, novel, safe and precise method has been developed for estimation of poorly water soluble drug Entacapone (Water Solubility-7.97e-02 g/l). Solubility of entacapone is increased by using 8M Urea as hydrotropic agent. There was more than 67 fold solubility enhanced in hydrotropic solution as compare with distilled water. The entacapone (ENT) shows the maximum absorbance at 378 nm. At this wavelength hydrotropic agent and other tablet excipients do not shows any significant interference in the spectrophotometric assay. The developed method was found to be linear in the range of 4-20 μg/ml with correlation coefficient (r2) of 0.9998. The mean percent label claims of tablets of ENT in tablet dosage form estimated by the proposed method were found to be 99.17±0.63. The developed methods were validated according to ICH guidelines and values of accuracy, precision and other statistical analysis were found to be in good accordance with the prescribed values. As hydrotropic agent used in the proposed method so this method is Ecofriendly and it can be used in routine quantitative analysis of drug in bulk drug and dosage form in industries.

Two plant essential oils (EOs), including those from Heracleum transcaucasicum and Heracleum anisactiss (Umbeliferae) were studied to detect the chemical constituents and evaluated for their antibacterial activities against Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli and Pseudomonas aeruginosa. The EOs of H. transcaucasicum and H.anisactis (Apiacae) were obtained by hydrodistillation from aerial parts of the plants. The chemical analyses of the EOs were performed by GC/Mass spectrometry (GC/MS). Myristicin was found to be the principal constituent in both EOs. The susceptibility tests of EOs were performed by agar disc diffusion technique against Gram-positive and Gram-negative bacterial strains. Eight components comprising 99.97% of the total essential oil of H. transcaucasicum and a total of three compounds accounting for 98.5% of the total oil composition of aerial parts of H. anisactis were identified, of which myristicin was the main compound in both EOs. The EOs of H. transcaucasicum and H. anisactis showed weak antibacterial property against Gram-positive strains of Staphylococcus aureus and Staphylococcus epidermidis with no measurable effect on Escherichia coli and Pseudomonas aeruginosa. Our GC-MS study revealed myristicin to be the major constituent of H. transcaucasicum and H.anisactis aerial parts. In spite of all the information available on the antibacterial properties of plants essential oils, we were not able to find significant antibacterial activity for both EOs.

Thermal behavior of some antidiabetic drugs such as pioglitazone hydrochloride (PTZ), rosiglitazone maleate (RGZ), glibenclamide (GBD) and glimepiride (GMP) has been studied. Thermogravimetric analysis (TGA), derivative thermogravimetry (DTG) and differential thermal analysis (DTA) techniques were used to study the thermal behavior of the drugs under investigation. Thermal analysis technique was used to obtain quality control parameters such as melting point 193.13 °C, 122.42 °C, 173.75 °C and 208 °C for PTZ, RGZ, GBD and GMP, respectively. The values of melting point of gave satisfactory results in comparison to that obtained by using the official method. Non-isothermal methods were employed to determine the activation energy values of the first stage of thermal decomposition. Comparison of the activation energy values suggests the following sequence of thermal stability: GMP > GBD > RGZ > PTZ. The results obtained are useful for the identification of these compounds and permitted interpretations concerning their thermal decomposition. Thermal stability of pharmaceutical compounds can be studied and compared by using thermal analysis techniques.

The aim of the present study was preparation, physicochemical characterization and performance evaluation of gold nanoparticles (GNPs) in radiotherapy. Another objective was the investigation of anti-bacterial efficacy of gold nanoparticle against E. coli clinical strains. Gold nanoparticles prepared by controlled reduction of an aqueous HAuCl4 solution using Tri sodium citrate. Particle size analysis and Transmission electron microscopy were used for physicochemical characterization. Polymer gel dosimetry was used for evaluation of the enhancement of absorbed dose. Diffusion method in agar media was used for investigation of anti-bacterial effect. Gold nanoparticles synthesized in size range from 57 nm to 346 nm by planning different formulation. Gold nanoparticle in 57 nm size increased radiation dose effectiveness with the magnitude of about 21 %. At the concentration of400 ppm, Nano gold exhibited significant anti-bacterial effect against E. coli clinical strains. It is concluded that gold nanoparticles can be applied as dose enhancer in radiotherapy. The Investigation of anti-bacterial efficacy showed that gold nanoparticle had significant effect against E. coli clinical strains.

Marrubium crassidens, a plant belonging to the family Lamiaceae, was studied for its volatile components present in the aerial parts of the plant during the flowering stage. The essential oil of the plant obtained through hydrodistillation of the dried plant material was assessed for its chemical composition by GC/MS and GC-FID analyses. Twenty-five compounds were identified, which constituted 94.3% of the total oil composition. The major components were identified as, m-tolualdehyde (23.3%), acetophenone (15.8%), nonacosane (13.1%), docosane (7.2%), o-tolualdehyde (4.1%), β-caryophyllene (3.8%) and caryophyllene oxide (3.4%). Non-terpenoids with 75.7% were the most abundant components of the essential oil. Overall, M. crassidens essential oil revealed to include rather higher proportions of non-terpenoid compounds compared with other species of genus Marrubium.

MSCs are multipotent progenitors which reside in bone marrow. They support hematopoietic stem cells homing, self renewal and differentiation in bone marrow. They can also differentiate into osteoblasts, adipocytes, chondrocytes, myocyates and many other tissues. In vivo, when trauma happens, MSCs operate cell renewal and migrate to the damaged tissues to regenerate that injury. In vitro, MSCs are able to proliferate and differentiate to a variety of cell lineages. This makes them a very hopeful tool for cell-based regenerative therapy for large bone defects, maxillofacial skeletal reconstruction, cardiovascular and spinal cord injury and so many other defects. The most important characteristic that make MSCs an excellent tool for cell replacement is their ability to escape from immune rejection. For therapeutic purposes they usually isolated from human bone marrow or fat and they should proliferate in order to reach an adequate number for implantation. Conventionally DMEM medium supplemented with 10% FBS is used for their expansion, but currently autologous platelet rich products are replaced FBS. Platelet granules contain so many growth factors that can support MSCs proliferation.

During long-term treatment with Levodopa, majority of patients with Parkinson's disease experience some abnormal motor problems including of Levodopa induced dyskinesia (LID) and wearing off. Incredible evidences suggest that serotonergic neurons compensate some functions of lost dopaminergic neurons in Parkinson's disease especially in advanced disease stages. Therefore, it has been postulated that serotonergic neurons are the major source for development of these unwanted effects. 5HT1A receptors are located on the serotonergic neurons and are involved in regulation of normal motor functions. With respect to the role of serotonergic projection in Parkinson's disease and importance of 5HT1A receptors in motor activity it seems that inactivation of these neurons by stimulation of 5HT1A receptors could provide benefits in treatment of Levodopa induced motor impairments.

Because mothers and fathers are more or less dissimilar at multiple HLA loci, mother considers her fetus as a semi-allograft. Mother's immune system may recognize paternal HLA as foreign antigen and may develop anti-paternal HLA antibodies and cytotoxic T lymphocyte. There are some mechanisms that modulate maternal immune responses during pregnancy, in order to make uterus an immune privileged site. This immunosuppression is believed to be mediated, at least partly, by HLA-G, non-classical class I human leukocyte antigen (HLA) molecule that is strongly expressed in cytotrophoblast and placenta. The major HLA-G function is its ability to inhibit T and B lymphocytes, NK cells and antigen-presenting cells (APC).Since HLA-G is expressed strongly at the maternofetal interface and has an essential role in immunosuppression, HLA-G polymorphism and altered expression of HLA-G seems to be associated with some complications of pregnancy, such as pre-eclampsia, recurrent misscariage and failure in IVF.This perspective discusses recent findings about HLA-G genetics, function, expression and polymorphism; and focus on HLA-G role in the etiology of recurrent miscarriage.

Eperisone Hydrochloride (EPE) is a potent new generation antispasmodic drug which is used in the treatment of moderate to severe pain in combination with Paracetamol (PAR). Both drugs are available in tablet dosage form in combination with a dose of 50 mg for EPE and 325 mg PAR respectively. The method is based upon Q-absorption ratio method for the simultaneous determination of the EPE and PAR. Absorption ratio method is used for the ratio of the absorption at two selected wavelength one of which is the iso-absorptive point and other being the λmax of one of the two components. EPE and PAR shows their iso-absorptive point at 260 nm in methanol, the second wavelength used is 249 nm which is the λmax of PAR in methanol. The linearity was obtained in the concentration range of 5-25 μg/mL for EPE and 2-10 μg/mL for PAR. The proposed method was effectively applied to tablet dosage form for estimation of both drugs. The accuracy and reproducibility results are close to 100% with 2% RSD. Results of the analysis were validated statistically and found to be satisfactory. The results of proposed method have been validated as per ICH guidelines. A simple, precise and economical spectrophotometric method has been developed for the estimation of EPE and PAR in pharmaceutical formulation.

The aim of this study was to investigate in vitro cytotoxic activity of four methanolic crude plant extracts against panel cell lines. Methanolic extracts were tested for their possible antitumor activity and cytotoxicity using the 3-(4,5-dimetylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay on six cancer cell lines; non-Hodgkin’s B-cell lymphoma (Raji), human leukemic monocyte lymphoma (U937), human acute myelocytic leukemia (KG-1A), human breast carcinoma (MCF-7 cells), human Prostate Cancer (PC3) and mouse fibrosarcoma (WEHI-164) cell lines and one normal cell line; Human Umbilical Vein Endothelial Cells (HUVEC). All species showed dose dependent inhibition of cell proliferation. IC50 values ranging from 25.66±1.2 to 205.11±1.3 μg/ml. The highest cytotoxic activity Chelidonium majus L> Ferulago Angulata DC> Echinophora platyloba DC> Salvia officinalis L, respectively. all extracts demonstrate promising cytotoxicity activity as a natural resource for future bio-guided fractionation and isolation of potential antitumor agents.

There are several reports about effects of Salvia spp. on CNS. The present experiment is undertaken to study effect of S. limbata, S. hypoleuca and S. macrosiphon on withdrawal syndrome in mice. Antinociceptive activities of aerial parts of Salvia spp. is investigated using hot plate method. In addition, the effect of its aerial parts on morphine dependence is investigated in mice. After induction of morphine dependency, different concentrations of plant extract are injected. To assess morphine withdrawal, naloxone (5 mg kg-1, i.p.) are injected into mice on the 5th day. Withdrawal syndrome is assessed by placing each mouse in a glass box 30 cm in height and recording the incidence of escape jumps for 60 minutes. A decrease in incidence of escape jumps is observed in morphine dependence mice. S. limbata and S. hypoleuca extracts produced a statistically significant inhibition of pain induced by hot plate latency at (500, 1000 and 1500 mg kg-1) i.p. A significant increase in pain threshold is observed after 30 and 60 minutes (p < 0.001). The activity was comparable to that of morphine (30 mg kg-1, i.p., p > 0.05). The antinociceptive activity increased up to 60 minutes. S. limbataand S. hypolecuca extracts produced statistically significant inhibition of pain and development of morphine dependence in mice.

Aflatoxin B1 (AFB1) is a potent mycotoxin which has been produced by fungi such as Aspergillus flavus and Aspergillus parasiticus as secondary metabolites due to their growth on food stuffs and induces hepatocellular carcinoma in many animal species, including humans. In the present study, the effect of AFB1 on STAT5A gene expression was investigated in bovine mammary epithelial cells using real time RT-PCR. Bovine mammary epithelial cells were seeded in a 24-well culture plate for three-dimensional (3D) culture in Matrigel matrix. After 21 days of 3D culture and reaching the required number of cells, cells were treated with AFB1 and incubated for 8 h. For real time PCR reaction, total RNA from the cultured and treated cells was extracted and used for complementary DNA synthesis. The expression of STAT5A gene was significantly down regulated by AFB1 in dose- dependent manner and led to the reduction of proliferation and differentiation of epithelial cells, which has direct effect in milk protein quantity and quality. According to the results, it seems that down regulation of STAT5A gene in AFB1-treated cells maybe due to DNA damage induced by AFB1 in bovine mammary epithelial cells.

The present study is aimed to evaluate phenolic profiles, cytotoxic activity and phytochemical screening of different extracts of Drynaria quercifolia leaves.

The dried and powder leaves were extracted with methanol at room temperature and the concentrated methanolic extract was fractionated by the modified Kupchan partitioning method to provide pet-ether, carbon tetrachloride, chloroform and aqueous soluble fractions. Phenolic profiles were determined by using Folin-Ciocalteau reagent, which results were expressed in gallic acid equivalent (mg of GAE/g of sample). Phytochemical properties of different extractives of plant materials were tested by the method of Trease and Evans. Brine shrimp lethality bioassay was used to investigate the cytotoxic potential of D. quercifolia.

The phytochemical screening revealed the potent source of different phytochemical constituents on different extractives including alkaloid, glycosides, tannin, saponins, proteins and amino acids, flavonoids, triterpenes, phenols, phytosterols and carbohydrate. In the determination of phenolic profiles, different extractives showed a significant content of phenolic compounds ranging from 103.43 -132.23 mg of GAE/g of extractive. Compared to vincristine sulfate different extractives of plant materials demonstrated moderate cytotoxic potential (having LC50 of 12.45 μg/ml, 13.02 μg/ml 15.83 μg/ml, 14.95 μg/ml and 7.612 μg/ml, respectively).

It is concluded from this study that D. quercifolia is an excellent source of phenolic content and phytoconstitutes as well as possesses moderate cytotoxic activity.

An accurate and precise UV spectrophotometric method with multivariate calibration technique for the determination of aripiprazole in pharmaceutical formulations has been described. This technique is based on the use of the linear regression equations by using the relationship between concentration and absorbance at five different wavelengths. The aripiprazole shows absorption maxima at 255 nm and obeyed Beer’s law in the range of 5-30 μg/mL. The results were treated statistically and were found highly accurate, precise and reproducible. This statistical approach gives optimum results for the eliminating fluctuations coming from instrumental or experimental conditions. It was concluded that the proposed method is simple, easy to apply, economical and could be used as an alternative to the existing spectrophotometric and non-spectrophotometric methods for the routine analysis of aripiprazole in pharmaceutical formulations.

The objective of this study was construction of recombinant hEGF-pPIC9 which may be used for expression of recombinant hEGF in following studies. EGF cDNA was purchased from Genecopoeia Company and used for PCR amplification. Prior to ligation, the PCR product and pPIC9 vector was digested with EcoRI and XhoI and ligated in pPIC9 vector and subjected to colony PCR screening and sequencing analysis. PCR amplification of EGF cDNA using recombinant hEGF-pPIC9 vector as template was concluded in amplification of 197bp fragment. Construction of recombinant hEGF-pPIC9 of EGf gene was verified by PCR and sequencing. Construction of Recombinant hEGF-pPIC9 was the primary stage for production and expression of EFG in the future study.

Gokhsuradi churna is an ayurvedic formulation, was investigate for antiurolithiatic activity. Calcium oxalate crystallization was induced by the addition of 0.01M sodium oxalate solutions in synthetic urine and nucleation method. The effect of Gokhsuradi Churna exhibited a concentration dependent inhibition of on calcium oxalate crystallization and nucleation. The present studies suggest that Gokhsuradi churna has a potential inhibition of calcium oxalate crystallization exhibited and nucleation. Gokhsuradi Churna showed potent antiurolethiatic activity.