International Journal of

Pharmaceutical and Clinical Research

e-ISSN: 0975 1556

p-ISSN 2820-2643

Peer Review Journal

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1. Evaluation of Antimicrobial Effect of Bauhinia Blakeana Flowers
Viji Saral Elezabeth D, Ramachandran P
This study was to evaluate the antimicrobial activity of petroleum ether extract of Bauhinia blakeana (Family-Fabaceae) flowers against selected microbes and it was obtained by continuous hot percolation method: Antimicrobial activity was evaluated by Agar well diffusion method. This investigation revealed the petroleum ether extract showing significant Antimicrobial activity against the selected microbes. Hence, we can conclude that the flower extract of Bauhinia blakeana was possessing Antimicrobial potential.

2. Sickle Cell Anemia in Relation to Total Homocysteine Levels and the Role of Anticoagulant Proteins
Mohammed Al-Nuzaily, Faisal Ali
Sickle cell disease (SCD) is a genetic disease characterized by hypercoagulable state and increased risk of thromboembolic events, a rare but significant complication of SCD. Protein C and S are natural anticoagulant proteins. Total homocysteine (tHcy) is an independent risk factor for venous thromboembolism and cardiovascular disease, and its level is therefore of interest in sickle cell disease. The aim of this study was to investigate the plasma levels of protein C and S levels and their relationship to homocysteine level in patients with sickle cell anemia compared to control subjects. In this study twenty patients (m=12, f=8) with sickle cell anemia, classified into sickle cell trait (Hb AS, n=15) and sickle cell disease (Hb SS, n=5), and twenty normal age-sex controls (m=12, f=8) were included. Protein C, S and homocysteine levels were measured using ELISA diagnostic kits technique. The data was statistically analysed by SPSS-17 and p values less than 0.05 were considered significant. Our results showed that mean tHcy levels were found to be significantly higher in patients (SS and/or AS) than in control group. No significant correlation was observed between tHcy with protein C or S.  The mean value of protein C and S within normal range and statistically not significant in patients compared to controls, but significantly decreased in Hb SS patients. In conclusion, sickle cell anemia is associated with mild elevated tHcy level which may contribute to increased risk of hypercoagulability and thromboembolic complications.

3. Non-Specific Immunostimulatory Capacity of Newcastle Disease Virus (NDV) and Suppression of Breast Cancer Cells
Ismaila Ahmed, Umar Ahmad, Yong Yoke K, Fauziah O
Newcastle disease virus (NDV) is an enveloped single stranded RNA virus that causes deadly infection to over 250 species of birds, comprising domestic and wild-type, thus resulting in substantial economic loss to poultry industry across the globe. NDV possesses several distinctive properties that make it an outstanding anti-cancer agent. In humans it is reported to have oncolytic and immune-stimulatory effects, precisely replicates in tumour cells while sparing normal cells and causes oncolysis. Although NDV has been extensively studied by researchers there is still need for a vigorous research on its potential use as a new treatment modality to cancer patients through a known process termed viroimmunotherapy. This paper deals with an overview of the research which has been carried out worldwide in the use of immune-stimulatory properties of NDV as an anti-cancer agent.

4. In vitro Synergistic effect of Cassia fistula leaves extracts with Fluconazole against clinical isolates of Candida albicans
Yogesh Bansal, Charulata, Devendra Singh Negi
The aim of the study was to evaluate the synergistic antifungal action of crude DMSO (Di methyl sulphoxide) & Petroleum ether leaves extract of Cassia fistula in conjunction with fluconazole against candidial species isolated from Candidasis patients. Correlation was observed between the leaves & fluconazole, therefore it was also found that the antifungal action of Cassia fistula leaves extract was enhanced (up to 3 times) in combination with fluconazole on fluconazole resistance Candida albicans strains isolated from patients with Candidiasis. The plant appears to play an important role in overcoming the azole resistant of Candidial strains. IC50 was determined by agar well diffusion and broth dilution method in the presence of fluconazole and leaves extracts of Cassia fistula in different solvents. Synergistic behavior of leaves extracts in Pet. Ether and DMSO extract of Cassia fistula with fluconazole was checked by broth dilution method.

5.  Bioanalytical Method Development and Validation of Niacin and Nicotinuric Acid in Human Plasma by LC–MS/MS
Bratati Roy, Bhupinder Singh, Anjana Rizal, C.P. Malik
An LC–MS method for the determination of Niacin and its metabolite Nicotinuric acid in human plasma was developed and validated. Sample preparation involved the solid phase extraction method. Chromatographic separation was performed on Phenomenex Gemini NX,5µm 4.6 mm x 100 mm column with the mobile phase consisting of Acetonitrile : (5 mM ammonium acetate buffer : Formic Acid::99.9:00.2 v/v) 70:30 v/v. The interface used with the API 3000 LC-MS/MS was a turbo ion spray in which positive ions were measured in MRM mode. The method was validated over the concentration range of 10.068-5002.086 ng/mL (NIC) and 10.157-5000.450 ng/mL (NIA). The recovery was 77.771% (NIC), 74.014 % (NIA) and the Lower imit of quantitation (LLOQ) was 10.068 ng/mL(NIC) and 10.157 ng/mL (NIA) . The intra- and inter-day precision of the method at four concentrations was 1.67-10.42% and 2.37-9.76% for NIC and 1.79-6.29% and 4.62-6.44% for NIA .Stability of compounds was established in a series of stability experiments. The method can be used for the simultaneous determination of Niacin and Nicotinuric acid in human plasma.

6. High Throughput Screening of Quorum Sensing Inhibitors Based Lead Molecules for Pseudomonas aeruginosa Associated Infections
Jha S K, Rashmi S, Shubhra, Singh H R
Quorum sensing (QS) is a process of cell-cell communication in bacteria by the use of signalling molecules that bind to the receptor protein and directly or indirectly affect transcription and translation. QS proteins may be used as a drug target for the inhibition of signalling pathway to control bacterial cell population. In present study the QS protein LasR of Pseudomonas aeruginosa was used as druggable target. Total fifteen inhibitors of LasR in 32 conformations were used for high-throughput computational docking. On the basis of docking score and glide score suitable inhibitors were identified. These selected inhibitors can be used as lead molecules for the designing of inhibitor based drugs. Lead molecules were further characterised by ADMET analysis, which include Lipinski’s rule, Jorgensen’s rule, blood-brain barrier penetration, Skin permeability, Human intestinal absorption and oral absorption. [(4E)-1-hydroxy-3-methylpyridin-4(1H)-ylidene]azinic acid] was found to be best inhibitor of LasR among all the inhibitors with best ADMET properties. In this study the above lead molecule was found to be a better alternative for the inhibition of bacterial population and in prevention in associated diseases. Present study focuses on the importance of structure based in silico drug design which takes less time and is cost effective.

7. Virtual Screening, Molecular Docking and Molecular Dynamics Studies For Discovery of Novel Vegfr-2 Inhibitors
Prabhu K, Manoj Kumar Mahto, Gopalakrishnan V. K
VEGFR-2 is considered as potential target for cancer therapy. In this work, the stability, binding mode between the VEGFR-2 protein and its ligand have been evaluated using the pharmacophore guided virtual screening (VS), molecular docking, and molecular dynamics (MD) simulations. The small molecule VEGFR-2 inhibitors were identified through virtual screening of chemical databases based on pharmacophore guided VS approach, that searches multi-conformer representations efficiently using PHASE module of Schrödinger. In addition, the molecular docking, using GLIDE module of Schrödinger; and molecular dynamics simulation, using GROMACS software were performed to study the interaction between the protein and the ligand. Molecular docking enables the extraordinary structural diversity of synthetic products to be harnessed in an efficient manner. The best six ligands (ZINC01056202, ZINC06091460, ZINC06091450, ZINC04107510, ZINC04623218, and ZINC81582433) with different scaffolds are selected from docking studies. VEGFR-2 and ligand complex was found to be stable at room temperature demonstrated by 1000 ps molecular dynamic simulation study using water as a solvent. The predicted inhibitors are quite novel compared with the known VEGFR-2 inhibitors. The work provides insight for molecular understanding of VEGFR-2 and can be used for development of anticancer drugs.

8. Evaluation of Analgesic Activity of Ethyl Acetate Extract of PergulariadaemiaForsk. Roots
Swapna.G, Joshi Anand.T, Dilip Raja.K
PergulariadaemiaForsk. Syn. Daemiaextensa(Asclepiadaceae) commonly known with the name of “dustaputeega” in telugu is a perennial twining herb, growing widely along the road sides of Andhra Pradesh state in India. Traditionally the plant is used to treat jaundice and it is used as anthelmintic, laxative, anti-pyretic and expectorant, and also used in infantile diarrhea. It is also used in treatment of malarial intermittent fevers. The present study was taken to evaluate the analgesic activity of Ethanolic extract of Pergularia daemia Forsk. Root. using eddy’s hot plate and heat conduction method. In eddy’s hot plate method the ethyl acetate extract showed significant analgesic activity at the doses of 200 mg/kg (p<0.05) and 400 mg/kg (p<0.01) as compared to control group, when analyzed statistically by Dunnet method, The result obtained show that the ethyl acetate root extracts of Pergulariadaemia (Forsk.) Chiov.possesses significant analgesic activity which confirms the traditional claims of the plant mentioned in Ayurveda.

9. Hepatoprotective Activity of Boerhavia diffusa Extract
Patel Monali, Verma Ramtej
Boerhavia diffusa Linn. (Nyctaginaceae), commonly known as ‘Punarnava’ is a perennial creeping herb widely studied and has a long history of uses by the tribal people and in Ayurvedic and Unani medicines. Present study was carried out to evaluate the ameliorative effect of the Boerhavia diffusa extract against carbon tetrachloride (CCl4) – induced hepatotoxicity in mice. Swiss albino female mice were cotreated with Boerhavia diffusa extract at three different doses (100, 200 and 300 mg/kg body weight/day) alongwith CCl4 (1/10 of the LD50  value). Serum marker enzymes, lipid peroxidation and histopathological analysis were carried out to evaluate the hepatoprotective effect of the Boerhavia diffusa plant extract. Cotreatment of Boerhavia diffusa extract (100, 200 and 300 mg/kg bw/day) along with CCl4  caused significant (p<0.05) decrease in the activities of serum marker enzymes (ALT, AST, ALP, ACP, LDH and γ-GT) as well as bilirubin contents and lipid peroxidation in a dose-dependent manner. Also cotreatment of Boerhavia diffusa extract significantly ameliorated CCl4 – induced histopathological changes and preserved the histoarchitecture of the liver tissue to near normal. Results of the present study indicate that the Boerhavia diffusa extract possess potent hepatoprotective activity against CCl4 – induced hepatic damage in Swiss albino mice which is mainly due to its antioxidative properties.

10. Comparision Efficacy of Hydrocortisone Acetate Cream in Patients
Manickam Tamilselvi, Ankanagari Srinivas
In a double-blinded, randomized, clinical trial lasting two weeks in 20 patients with skin infections (Eczema, Dermatitis, and pruritus), inflammatory and Pruritic manifestations of corticosteroid responsive dermatoses Pruritus, Eczema and Atopic Dermatitis were treated and compared between 1% w/w hydrocortisone cream and reference 1% w/w cutisoft hydrocortisone acetate cream and reductions of the basic criteria, i.e. itching, erythema and scaling, were evaluated.  Effects were compared using Visual Analogue Scale, Patients compliance and Physician Global Evaluation of efficacy. All treatment regimens significantly reduced itching, erythema and scaling after 4 visits. The patients were assessed on day 01 (Visit 1), day 05 + 01 (Visit 2), day 10 + 01 (Visit 3) and on day 14 + 01 (Visit 4) for the analyses of infection. The basic criteria scores were decreased from visit 1 to visit 4. All patients were well tolerated. The results of therapeutic outcome proved the better results for test cream over reference cuti soft cream, the statistical analysis based on Generalized Linear model shown that both the products are clinically equivalent in terms of anti-inflammatory effect of hydrocortisone cream in this study.

11. Vitamin D and Glycemic Control in Impaired Fasting Glycemia
Vinodhini V.M, Arun Krishnan A, Ebenezer William W.
Impaired Fasting Glycemia (IFG) is defined as elevated fasting plasma glucose concentration ≥100mg/dl and <126 mg/dl. IFG is considered to be an intermediate state in the development of diabetes and cardio vascular diseases. Vitamin D and calcium homeostasis may also play a role in the development of type 2 Diabetes Mellitus (DM). The aim of the study was to correlate the levels of serum 25hydroxy vitamin D with the glycemic parameters in a group of subjects with Impaired Fasting Glycemia. The study included 30 participants with IFG. Fasting Blood Glucose (FBG) & Post-Prandial Blood Glucose (PPBG) were estimated by Glucose Oxidase Peroxidase method, Glycated Hemoglobin (HbA1c) levels were estimated by Ion exchange HPLC method using Biorad analyzer. Serum Vitamin D levels were estimated by competitive ELISA kit (DLD Diagnostika GmbH).  Correlation between the levels of Serum 25 hydroxy Vitamin D with FBG, PPBG and HbA1c was analysed by Pearson’s Correlation. There was a negative correlation between Vitamin D levels and FBS, PPBS and HbA1c in subjects with IFG. The correlation between Vitamin D levels and FBG and the correlation between Vitamin D and HBA1C in subjects with IFG was found to be statistically significant. The results of this study have shown that IFG is associated with decreased levels of 25 (OH) Vit D.

12. Cloning and Expression of iceA1 Gene of Helicobacter pylori.
Seyed Hossein Zendehbad, Mohammad Javad Mehran., R. Senthil Kumar
Helicobacter pylori, a gram-negative, motile, spiral-shaped bacterium, have been established as the etiologic agent of chronic gastritis and belongs to phylum Proteobacteria. The gene iceA (induced by contact with epithelium) is also proved to be an important factor for the pathogenecity. The iceA1 gene is usually expressed when Helicobacter comes in close contact with the epithelial cells of the gastric lining. Moreover its presence has also been identified in those patients suffering from peptic ulcers. Due to the fact of its involvement in the contact to the epithelial layers, iceA gene has been focused primarily in the disease diagnosis. Characterization and cloning of this gene could be a potential mode of vaccine development towards this gastric cancer. And moreover the antibiotic based drugs are of limited use. Novel methods are being developed for the production of antibodies to specific antigens and thus helping in the process of development of protein based vaccines. iceA gene was isolated and ligated into pTZ57R/T cloning vector. The ligated product was then cloned into DH5α strain and allowed to propagate. The plasmids thus cloned were purified and later expressed for the gene of interest in an expression vector. The proteins specific to the gene of interest was then isolated and purified. This proteins purified can in turn be used for protein based vaccines.

13. Atomic Absorption Spectroscopic Determination of Few Major and Trace Elements in Nature’s Finest Medicine (Wheatgrass Juice Powder) Triticum  aestivum L. and their Possible Correlation With Reported Therapeutic Activities: Part-I
Sharma Vivek, Sharma Navdeep, Bano Anisha, Kumar Sanjeev, Dhaliwal Harcharan Singh
The young wheat plant (Triticum aestivum L.) belongs to the family Poaceae is referred as wheatgrass. It provides all the important nutrients for a healthy and rejuvenating body and acts as an efficient source of energy. The extensive research on wheatgrass is already mentioned in Ayurveda, different herbal system of medicines and different pharmacopoeias of the world. It possesses number of pharmaceutical properties, in which some have already been known and many more are still to be identified. In the present study the major and trace element contents like (Zn, Cu, Mn, Fe, Mg, K and Ni) were first time comparatively analysed in the three samples viz.: Wheat Grass Juice Powder (WGJP), Wheatgrass Fibre Powder (WGFP) and Dry Wheatgrass Powder (DWGP) of Triticum aestivum L. from Northern India with the help of Atomic Absorption Spectrophotometer (AAS). The main purpose of the exploration is to document confirmation and spreading the benefits of wheatgrass to daily users, practitioners and also for the researchers worldwide by correlating the properties of these elements with reported therapeutic activities.

14. The Effect of an Indigenous Drug on Abnormal Folate Metabolism and Intellectual Disability: A Study of 5,10-Methylenetetrahydrofolate Reductase Activity
Rinki Kumari, Aruna Agrawal, O.P.Upadhyay, K. Ilenago, G.P.I.Singh, G.P.Dubey
Methylenetetrahydrofolate reductase, (MTHFR) is a key enzyme that is required to metabolize folate and is essential for normal development of central nervous system. Mutation in the MTHFR gene may result in elevated plasma homocysteine level. MTHFR C677T is most common polymorphism associated with impairment of cognitive function. In the present study, we examined the relationship between 677C>T MTHFR gene polymorphisms, homocysteine (tHcy) and the effect of test drug in the level of tHcy in the treated intellectual disability children (ID). Fifty patients of the both genders (male 31, female 19) with an age range of 8 to12 years with ID (cases) were selected from Pediatric O.P.D of S.S. Hospital I.M.S, B.H.U. ID children were clinically diagnosed according to the DSM V screening system. MTHFR C677T polymorphism was detected in the normal control vs.ID children by PCR-RFLP is using Hinf I enzyme. A total 30 out of 50 ID children were treated with the test formulation and remaining 20 ID children were placebo. Plasma homocysteine levels were measured by HPLC at initial, 3 & 6 months of study. CT of MTHFR C677T was higher in ID children than controls, although it did not show statistical significance (p=0.1) and the child T677T MTHFR genotype (TT) was found 4 % in ID children In the treated group, level of homocysteine dropped while increased in the placebo after six months of treatment. In treated group ID children, CC with genotype the level of tHcy dropped from 9.06 to 8.21; CT 11.92 to 10.15 whereas in placebo no such change was observed. Therefore, the present study showed MTHFR C677Tpolymorphism associated with modifying level of tHcy. The result indicated that ID children with the MTHFR C677T gene showing the highest level of tHcy, when treated with test formulation exerted reduction in tHcy concentration suggesting improved cognitive function due to homocysteine re-methylation. The present study also suggested an association of high degree of genetic heterogeneity because of their mutation in ID children. It is concluded that the test formulation has the potentiality to reduce the risk of MTHFR C677T gene mutation because of its activity and thus, improved the cognitive ability of ID children.

15. Study on Production of Extracellular Amylase from Bacillus subtilis Strain KPA under Mild Stress Condition of Certain Antimicrobials
Khusro A, Aarti C, Preetam Raj JP, Panicker SG
The present study was investigated to improve Bacillus subtilis strain KPA for extracellular amylase production under mild stress condition of certain antimicrobials. The novel isolated bacterial culture was exposed to different concentrations of Allium sativum, ampicillin and Mercuric chloride at their sub- MIC values for 24 to 72 h. Exposure of Allium sativum and ampicillin to strain KPA did not give improved extracellular amylase production. Allium sativum and ampicillin at the concentration of 250 µ l and 100 µ l were able to induce strain KPA for amylase production of 1896.257 U/ml/min and 1740.976 U/ml/min respectively after 48 h of incubation. On the other hand Mercuric chloride at the concentration of 250 µ l was able to improve strain KPA even after 24 h of incubation for maximum enzyme production (2024.136 U/ml/min) compared to the enzyme produced (1854.099 U/ml/min) by the parental strain. The extracellular enzyme produced by strain KPA under mild stress condition of certain antimicrobials tested here were 2-3 fold higher than that of previously isolated Bacillus strains. The results clearly conclude the role of novel isolated strain in various industries such as textile, paper, pulp, food etc. due to enhanced production of amylase under mild stress conditions.

16. The Impact of Clinical Pharmacists’ Interventions on Drug Related Problems in a Teaching Based Hospital
Muhammad Umair Khan, Akram Ahmad
Drug related problems are considered as a major problem associated with hospitals. Many studies have been conducted in past in evaluating the role of clinical pharmacist in reducing this prevailed problem, however only few have assessed the impact of clinical pharmacist intervention in addressing this issue. The objective of this study was to evaluate the impact of clinical pharmacists in reducing drug related problems in Pakistan A prospective, observational and interventional study was conducted in a teaching based hospital to identity drug related problems and their causes. Interventions were also designed to address these issues. Classification of drug related problem was done on the basis of PCNE system. Data were then entered into Microsoft excel (2010) for descriptive analysis. A total of 373 profiles were reviewed in which 184 profiles had drug related issues.  A total of 147 drug related problems were identified in which major issue was related to adverse drug reaction (n=61, 41.5%). The total number of causes which lead to these problems were 161, of which dosing error was found to be more prevalent (n=68, 42.2%). Out of 161 recommendations given by clinical pharmacists, 86.33% (n=139) were successful in solving the problem while 6.83% of recommendations were termed ineffective as they failed to address their respective issues. The clinical pharmacists play an important role in healthcare settings by reducing the burden of drug related problems effectively. Their contribution has promoted positive changes in healthcare setup of Pakistan.


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