Open Access Original Research Article

Effects of Oxidation on Protein-Nanoparticle Interactions

Valdez R. Rahming, Md. Abul Fazal

Journal of Pharmaceutical Research International, Page 172-185
DOI: 10.9734/BJPR/2014/5959

Aims: Upon entrance into the blood stream most nanoparticles bind to an array of proteins forming a “protein corona”. Fibrinogen is the second most abundant blood protein and has been reported to bind to a variety of nanoparticles including metal oxides, polymeric nanoparticles and carbon nanotubes.
Study Design: Study the effects of oxidation on the binding interactions between human serum fibrinogen and magnetic iron (III) oxide nanoparticles.
Place and Duration of Study: Department of Chemistry, College of St. Benedict, 37 South College Avenue, St. Joseph, MN 56374, U.S.A., between June 2011 and May 2012.
Methodology: Spectroscopic techniques (UV-Vis, IR, fluorescence, and circular dichroism) were used to study the binding interactions of magnetic nanoparticles with human serum fibrinogen and the effects of protein oxidation on its binding affinity.
Results: Magnetic nanoparticles (MNP) formed stable complex with fibrinogen under physiological conditions. The binding constants (Ka) were determined as 1.91 (± 0.14) x106 M-1 and 1.06 (± 0.09) x106 M-1 at 300 K and 310 K respectively. The secondary structure of the protein was slightly affected by the formation of fibrinogen-MNP complex. When the protein was oxidized with metal catalyzed oxidation (MCO) system, significant changes in the protein structure was detected leading to decreased binding affinity for MNP.
Conclusion: Metal catalyzed oxidation of fibrinogen significantly affects its binding interactions with magnetic iron (III) oxide nanoparticles.

Open Access Original Research Article

In Vivo Assay of Analgesic Activity of Methanolic and Petroleum Ether Extracts of Manilkara zapota leaves

Manirujjaman ., Farhana Sultana, Md. Arifur Rahman Chowdhury, Md. Tanjir Hossain, Md. Imran-Ul-Haque

Journal of Pharmaceutical Research International, Page 186-191
DOI: 10.9734/BJPR/2014/5941

Aims: The main objective of this work was to observe the analgesic activity of Manilkara zapota (leaves) on mice.
Study Design: The Present study was designed to observe pharmacological activities of the crude extracts of the plant Manilkara zapota (leaves). The study protocol consisted of Cold extraction at room temperature of the whole plant with distilled methanol. Afterwards, Filtration of the crude Methanolic and Petroleum ether extracts by using the Markin cotton cloth and subsequently through the filter paper and solvent evaporation. Finally, screening of analgesic activity of crude extracts on Swiss Albino mice.
Place and Duration of Study: School of Science and Engineering, Department of Pharmacy, Southeast University, Bangladesh, January 2011 to August 2012.
Methodology: The analgesic activity was investigated for its peripheral pharmacological actions by using acetic acid-induced writhing test in mice.
Results: The Methanolic and Petroleum Ether extracts, at the dose of 200 mg/kg body weight, displayed 96.82% & 94.27% pain inhibition which was significant (p<0.001) compared to control. These results indicate that the extracts possess significant analgesic activity.
Conclusion: This study suggests that the Methanolic and Petroleum Ether extract of Manilkara zapota leaves have analgesic activity in a dose dependent manner which supports it’s as an analgesic drug in folk medicine.

Open Access Original Research Article

Leaf Extracts of Vernonia amygdalina Del. from Northern Ghana Contain Bioactive Agents that Inhibit the Growth of Some Beta-lactamase Producing Bacteria in vitro

Oseni Lateef Adebayo, Abugri James, Sumabe Balagra Kasim, Onilimor Peter Jagri

Journal of Pharmaceutical Research International, Page 192-202
DOI: 10.9734/BJPR/2014/3200

Aims: The current study was aimed at evaluating the phytochemical profile and in vitro bacteria growth inhibitory potential of different solvent leaf extracts of V. amygdalina from northern Ghana.
Study Design: Different solvent extracts of the plant were quantitatively and qualitatively evaluated for phytochemicals. In vitro bacteria sensitivity assay of the extracts was evaluated using some beta-lactamase producing bacteria as test microbes.
Methodology: Ethanolic, methanolic, petroleum ether and aqueous leaf extracts of Vernonia amygdalina were studied in vitro for growth inhibition against beta-lactamase producing bacteria such as Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae and Pseudomonas aeruginosa using agar well diffusion method. Saponins, flavonoids, glycosides, alkaloids, tannins, phenolics, reducing sugars, anthracenosides, terpenes and phytosteroids were determined qualitatively and quantified.
Results: All the phytochemicals tested were found to be present in both the methanolic and ethanolic leaf extracts. The aqueous extract showed the presence of saponins, reducing sugars and anthracenosides. Glycosides, flavonoids, saponins and alkaloids were the only groups of phytochemicals found in the petroleum ether extract. The methanolic extract showed the greatest amount of saponins (14.23%), flavonoids (2.15%), alkaloids (7.49%), tannins (5.4%), terpenes (10.20%) and phenolics (8.24%). The methanolic extract at concentration of 4 mg/ml showed growth inhibitory activities against all the test organisms with zone of inhibitions ranging from 16.00+0.50 (against E. coli) to 20.50+0.03 mm (against S. aureus). The ethanolic extract showed activity against only two of the test organisms viz. 23.00+0.33 mm against P. aeruginosa and 12.00+0.00 mm against S. aureus at similar concentration. All test organisms were resistant to both aqueous and pet ether extracts.
Conclusion: The antibacterial activities of the methanolic and ethanolic extracts were significant (P < 0.05) and may be mediated by the presence of saponins, flavonoids, tannins, terpenes and alkaloids. Results from present study corroborate previous findings and also presents methanolic leaf extract of the plant as a credible candidate for the discovery of new phytotherapeutic agents against the beta-lactamase producing bacteria tested.

Open Access Original Research Article

Comparative Effect of Carbimazole, Citrus sinensis, Glycine Max and Levothyroxine on Blood Glucose Levels and Body Weight Changes

Okon Uduak Akpan, Udobang John Akpan, Ologhaguo Macstephen Adienbo, Ani Elemi John

Journal of Pharmaceutical Research International, Page 203-214
DOI: 10.9734/BJPR/2014/5665

Aims: The role of thyroid hormones as important mediator of glucose metabolism and body weight dynamics had long been established. This study was therefore designed to evaluate the effect of standard thyro-active drugs in comparison to crude nutritional extracts, on blood glucose level and body weight changes.
Methodology: Albino wistar rats weighing between 100 - 150g were randomly assigned to seven groups of seven rats each. Group 1 served as control, while groups 2-7 were orally administered Fresh orange juice(FOJ) (1500mg/kg), Fresh soybean(FSB) (0.01mg/kg), levothyroxine (LVT) (0.01mg/kg), carbimazole (Carb) (0.01mg/kg), FSB+LVT and FOJ+Carb respectively once daily for twenty eight days. Weekly weight records were taken and the difference calculated as weight changes. The animals were sacrificed and blood samples collected by cardiac puncture. Serum was obtained by standard procedure and used for blood glucose estimation.
Results: Separate treatment with both FOJ and FSB significantly (P<0.05) increased the blood glucose levels and body weight compared to control. Levothyroxine significantly (P<0.05) increased the blood glucose levels but significantly (P<0.05) decreased the body weight compared to the FOJ treated group. Carbimazole significantly (P<0.05) decreased the blood glucose levels compared to the control, FOJ and FSB treated groups. FSB+LVT treatment significantly (P<0.05) decreased the blood glucose levels compared to the LVT group and also significantly (P<0.05) decreased the body weight compared to the control, FOJ and FSB groups. FOJ+Carbimazole combination treatment significantly (P<0.05) decreased the blood glucose levels compared to LVT treatment and also significantly (P<0.05) decreased the body weight compared to the FOJ and FSB groups. FOJ significantly reduced T4 and T3 levels compared to control. FOJ, Carb and FOJ+Carb significantly (p<0.05) reduced T4 level compared to control.
Conclusion: The acclaimed antithyroid activity of FOJ and FSB did not appear to directly correlate with that of carbimazole, a standard antithyroid drug, considering their effect on blood glucose level and body weight changes. However their combination treatment with Carb and LVT respectively, showed similar pattern with carbimazole.

Open Access Original Research Article

Comparative Studies on Dissolution and Bioavailability of Tamoxifen Citrate Loaded Binary and Ternary Solid Dispersions

Dalia S. Shaker, Mohamed K. Ghorab, Mohamed S. Teiama

Journal of Pharmaceutical Research International, Page 215-229
DOI: 10.9734/BJPR/2014/6072

Aims: This study focused on comparing binary and ternary solid dispersions (SD's) of water-insoluble Tamoxifen citrate (TMC) regarding drug dissolution and oral bioavailability. Basically, the enhanced dissolution of this drug by water-soluble polymer has not yet been reported in literature.
Study Design: In vitro and In-vivo characterization of Tamoxifen citrate loaded binary and ternary solid dispersion systems.
Place and Duration of Study: Department of Pharmaceutics and Industrial Pharmacy, Helwan University, Cairo, Egypt between June 2012 and June 2013.
Methodology: Amorphous SD's of TMC with two hydrophilic polymers, polyethylene glycol 6000 (PEG 6000) and Methyl cellulose (MC), were prepared by melting method. Binary SD's of TMC with PEG of different weight ratios were prepared. MC was used as third component to prepare ternary SD's. Physicochemical properties of SD's were characterized by FTIR, DSC, and In-vitro drug release in comparison with physical mixtures and the drug alone. Oral bioavailability of the optimized SD formula was compared with that of free TMC in rats.
Results: Infrared spectroscopic studies suggested no interaction between TMC and polymers rather than H-bond formation. A remarkably improved dissolution of drug from the ternary solid dispersion systems when compared to the binary solid dispersion systems was detected. On the basis of % dissolution efficiency (% DE), the SD composed of PEG: TMC: MC in a ratio 4:1:2 w/w/w was selected as the optimized SD. The in-vivo studies showed extremely significant higher values of Cmax (P<.05), AUC0-24 (P<.05) and significantly (P<.05) lower values of Tmax exhibited by SD compared with free TMC.
Conclusion: Highly enhanced TMC dissolution and bioavailability exhibited by PEG: TMC: MC ternary solid dispersion in a weight ratio 4:1:2 were promising to improve the therapeutic potential of TMC.

Open Access Original Research Article

Effect of Andrographis serpyllifolia Leaves Extract on Experimentally Induced Typhoid Using Salmonella Typhi

Shyam Sundar Gupta, Joni Sharma, G. Rajesh Kumar, Garima Pandey, P. K. Mohapatra, A. K. S. Rawat, Ch. V. Rao

Journal of Pharmaceutical Research International, Page 230-239
DOI: 10.9734/BJPR/2014/5804

Aims: The aim of the study was to evaluate the effect of the standardized ethanolic extract of Andrographis serpyllifolia leaves on experimentally induced typhoid.
Study Design: Single dose of 1 ml Salmonella Typhi (106 CFU/mL) was administered orally to rats to induce typhoid in rats. Blood culture test confirmed typhoid infectioned rats received orally the ethanolic extract of Andrographis serpyllifolia at dose levels of 200 and 400 mg/kg twice daily for 10 days, respectively and control animals received physiological saline.
Place and Duration of Study: CSIR-National Botanical Research Institute (NBRI), Lucknow, between December 2011 and June 2013.
Methodology: Leaves of Andrographis serpyllifolia was extracted with ethanol and concentrated on rotavapour. Single dose of 1 ml S. Typhi (106 CFU/mL) was administered orally to rats with the help of orogastric tube to induce typhoid in rats. After seven days, typhoid confirmed rats received the standardized extract subsequently subjected to in vitro antimicrobial susceptibility test.
Results: The treatment with ethanolic extract of Andrographis serpyllifolia at dose level of 200 mg/kg showed 75.0% to 87.5% protection and 100% protection observed at higher dose of 400 mg/kg on widal, blood culture and typhidot test respectively. Biochemical test carried out on blood culture isolates confirmed the presence or absence of S. Typhi. A. serpyllifolia extract at a concentration of 1.50 mg/disc showed antimicrobial activity susceptibility against S. Typhi.
Conclusion: Andrographis serpyllifolia leaves extract showed antimicrobial activity against S. Typhi and accomplished the extract of A. serpyllifolia is recommended for clinical applications in the treatment of typhoid.

Open Access Original Research Article

Influence of Azithromycin Treatment on Hepatic Lipid Peroxidation and Antioxidant Defence Systems of Rats

Ebenezer Tunde Olayinka, Ayokanmi Ore

Journal of Pharmaceutical Research International, Page 240-256
DOI: 10.9734/BJPR/2014/3853

Aim: Azithromycin is a semisynthetic macrolide antibiotic commonly indicated for use in middle ear, upper and lower respiratory tract infections, bronchitis, and community-acquired pneumonia with a well-established safety and efficacy profile. The antibiotic is usually well tolerated; however, it has been associated with rare cases of progressive cholestatic hepatitis and fulminant hepatic failure. This study was therefore designed to examine the effect of two doses of Azithromycin; 5.6 mg/kg body weight, (b.w.) and 11.2 mg/ kg b.w. on some hepatic and renal function indices, oxidative stress and antioxidant defense system in rats.
Study Design: Toxicological, Histological and Biochemical study.
Place and Duration of Study: Biochemistry Unit, Department of Chemical Sciences, Faculty of Natural Sciences, Ajayi Crowther University, Oyo, Nigeria between March 2012 and April 2012.
Methodology: Thirty rats (Wistar strain) weighing between 180 – 200 g were randomly assigned into three treatment groups of ten animals each; Group I (Control) did not receive any drug, Group II received 5.6 mg/ kg b.w. and Group III received 11.2mg/ kg b.w. Azithromycin by oral gavage twice daily for seven days.
Results: Urea, Creatinine and Bilirubin levels were significantly (p=0.05) elevated in the plasma of the rats that received 5.6 mg/ kg b.w. and 11.2mg/ kg b.w. by 20.0% and 35.0%, 78.0% and 130.0%, and 14.0% and 47.4% respectively when compared to the control. Activities of some marker enzymes; aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and Gamma-glutamyl transferase (GGT) were also significantly increased (p=0.05) in the plasma of treated animals by 19.0% and 36.6%, 27.0% and 36.4%, 16.0% and 23.5%, and 43.9% and 80.5% respectively. Also, there was a significant increase (p=0.05) in plasma Triglycerides, Total cholesterol, HDL- and LDL-cholesterol in the treated animals by 20% and 31.2%, 19% and 35.4%, 24% and 74.3%, and 33% and 35.2% respectively. Furthermore, the two doses of Azithromycin significantly (p=0.05) reduced the levels of hepatic Ascorbic acid, Glutathione (GSH) and activities of Glutathione-S-transferase (GST) by 35%, 66.3% and 34% and 56.1%, 45% and 50%, respectively. In addition, there was a significant decrease in the activities of hepatic Catalase (CAT), and Superoxide dismutase (SOD) by 67.4% and 43.0%, and 43% and 50% respectively in the two treated group. These were accompanied by a significant increase (p=0.05) in hepatic lipid peroxidation (LPO) by 81% and 91.6% respectively in the treated groups. The histology of the liver revealed sinusoidal and portal congestion and a mild periportal cellular infiltration by mononuclear cells, by Zithromax 500®. Likewise, kidney histopathology revealed severe cortical congestion and hemorrhage and few tubules containing protein casts in the treated group.
Conclusion: In conclusion, the administration of 5.6 mg/ kg b.w. and 11.2mg/ kg b.w. of Azithromycin induced marked renal and liver damage, oxidative stress and altered the antioxidant status in rats.

Open Access Original Research Article

Wound Repair and Anti-inflammatory Potential of Fumaria indica in Excision Wound-induced Rats

Garima Pandey, G. Rajesh Kumar, Shyam Sundar Gupta, S. K. Ojha, Ch V. Rao

Journal of Pharmaceutical Research International, Page 257-266
DOI: 10.9734/BJPR/2014/5387

Aims: Objective of the study was to investigate the wound contraction and anti-inflammatory activity of the 50% ethanolic extract of Fumaria indica (Hausskn.) Pugsley (Fumariaceae) by excision wound model and estimation of pro-inflammatory and anti-inflammatory cytokines.
Study Design: Prospective.
Place and Duration of Study: Department of Pharmacognosy and Ethnopharmacology, CSIR-National Botanical Research Institute, Lucknow, India. December 2012 to May 2013.
Methodology: Dried powdered whole plant of Fumaria indica was extracted with 50% ethanolic extract. The extract was subjected to HPTLC fingerprinting, DPPH free radical scavenging and antibacterial activities. Further, 10% F. indica ointment was tested for its wound contraction, pro-inflammatory and anti-inflammatory potentials.
Results: The 50% ethanolic extract showed presence of ellagic acid, ferulic acid andquercetin. The IC50 was 0.11mg/mL and significant antibacterial activity was observed against S. aureus and E. coli. The 10% F. indica ointment applied topically to the wound area reducedits size from 500 mm2 to 40 mm2 by the end of 9th day. These results were comparable to the effect of 0.2% nitrofurazone. The extract further showed a reduction in the release of pro-inflammatory cytokines (TNFα and IL-6) and an increase in anti-inflammatory cytokine IL-10.

Open Access Original Research Article

In-vitro Evaluation of Cytotoxic and Anthelmintic Activity of Luffa acutangula, Luffa aegyptiaca and Momordica cochinchinensis

Md. Mosiqur Rahman, Ashik Ahmed, Saibal Saha Sunny, Md. Samiul Huq Atanu, Abdullah Faruque, Md. Sohel Rana

Journal of Pharmaceutical Research International, Page 267-277
DOI: 10.9734/BJPR/2014/4613

Aims: The study was designed to investigate cytotoxic and anthelmintic activity of aerial parts of Luffa acutangula (L.) Roxb. (Family: Cucurbitaceae, locally known as ‘Jhinga’), Luffa aegyptiaca Mill. (Family: Cucurbitaceae, locally known as ‘Dhundul’) and Momordica cochinchinensis (Lour.) Spreng. (Family: Cucurbitaceae, locally known as ‘Kakrol’) extracted with various solvents (petroleum ether & methanol).
Study Design: Determination of cytotoxic and anthelmintic activity of aerial parts of three (Cucurbitaceae family) Bangladeshi plants.
Place and Duration of Study: Department of Pharmacy, Jahangirnagar University, Savar, Dhaka-1342. Performed between November 2011- September 2012.
Methodology: The cytotoxic activity was evaluated by Brine Shrimp lethality bioassay and anthelmintic activity by in-vitro test using earth worm Pheretima posthuma (Annelida) as test animals.
Results: In Brine Shrimp lethality bioassay, methanol extract of M. cochinchinensis and L. aegyptiaca were found to be highly toxic to Brine Shrimp nauplii, having LC50 of 1.91±0.79 μg/ml and 3.97±0.61 μg/ml respectively. The three methanol extracts of aerial part of L. acutangula, L. aegyptiaca and M. cochinchinensis showed moderate anthelmintic activity. 50mg/ml concentration of methanol extract of M. cochinchinensis showed maximum activity showing death in test animals at 43±1.3 min which is comparable to the standard (Piperazine Citrate, 10 mg/ml) which killed the test animal at 38 ± 0.63 min.
Conclusion: Further studies are suggested to be undertaken to understand the underlying mechanism of the observed cytotoxic and anthelmintic activity of these three Bangladeshi (Cucurbitaceae family) plants.

Open Access Original Research Article

Analysis of Amantadine Hydrochloride-Phenyl Isothiocyanate Complex in Bulk and Pharmaceutical Dosage Forms by RP HPLC-PDA Method

Sri Kalyani Vemuri, N. Ramya Krishna, M. J. Mohammad, Buchi N. Nalluri

Journal of Pharmaceutical Research International, Page 278-288
DOI: 10.9734/BJPR/2014/6374

Aim: The aim of the present investigation was to perform the precolumn derivatization of Amantadine Hydrochloride (AMT) with phenylisothiocyanate and to develop a RP-HPLC-PDA method for the quantification of Amantadine Hydrochloride-phenylisothiocyanate (AMT-PITC) complex in bulk and dosage forms which is rapid, sensitive and economical.
Study Design: Method development and Validation study.
Methodology: A Phenomenex C18 RP column of 250 x 4.6mm dimensions and 5µm particle size with mobile phase containing water and acetonitrile (40:60% v/v) was used at isocratic mode binary pump and eluent was monitored at 273nm.
Results & Discussion: The retention time of AMT-PITC complex was 6.3 min. The developed method showed a good linearity in the concentration range of 10-50µg/mL with a correlation coefficient >0.998. The recoveries ranged between 95-105% with a Relative Standard Deviation of (RSD) < 2%.
Conclusion: The developed method was validated as per ICH guidelines and successfully used for quantification of AMT by derivatization with PITC. The method was found to be rapid, specific and accurate.