Open Access Method Article

A Comparison of the Humoral Immune Response Induced by a Recombinant Human Protein in Wild Type Mice and in Transgenic Mice Expressing the Protein

Britta Granath, Jan Holgersson, Karin Cederbrant, Ann Lövgren, Nina Brenden

Journal of Pharmaceutical Research International, Page 2511-2524
DOI: 10.9734/BJPR/2014/10923

Aim: The aim of this work was to investigate the correlation between anti drug antibody (ADA) induction and how different manufacturing processes of biopharmaceuticals affect the immunogenicity of the protein. This was done by testing four different batches of the same recombinant human protein in transgenic (Tg) mice.
Methodology: Wild type (Wt) and human protein-transgenic (Tg) mice were challenged by repeated subcutaneous injections of four batches of a drug candidate protein, obtained by different purification methods. Differences between drug-specific IgG1, IgG2a, IgG2b, IgG3 and IgM antibody patterns produced in Tg vs. Wt mice were investigated and compared to the plasma cytokine profiles. A conventional ELISA was used as a reference method for ADA detection.
Results: ADA responses detected in Tg mice were mainly of the IgG1 subclass and occurred only in significant response to the batch containing the highest level of proteins originating from the recombinant host cells. Wt mice, on the other hand, showed a combined IgG1/IgG2b response to all drug batches, except to the batch with the highest purity. The most pure batch failed to induce significant ADA in both Wt and Tg animals, suggesting host cell derived impurities to be a strong contributing factor to the antibody responses observed.
Conclusion: Thus, an isolated IgG1 response in drug-tolerant Tg mice may serve as a potential biomarker of an immunological reaction to process-related impurities of the protein drug. In contrast, a combined IgG1/IgG2b-profile, as observed in immunoreactive Wt mice, more likely reflects a xeno-response.

Open Access Original Research Article

Safety of Prosopis juliflora (Sw.) DC. (Fabaceae) and Entada leptostachya Harms (Leguminosae) Extract Mixtures Using Wistar Albino Rats

D. Kimani, P. G. Kareru, H. L. Kutima, F. K. Njonge, G. C. Nyagah, J. M. Karanja, K. Mutembei, P. Githira, G. Mercy

Journal of Pharmaceutical Research International, Page 2475-2483
DOI: 10.9734/BJPR/2014/10993

Aim: Many medicinal plants have been used traditionally in treating ailments in humans and animals. However, for most of herbal remedies, no scientific toxicity profiles exist in literature. In this study, the safety profile of an herbal extract mixture containing Entada leptostachya (EL) and Prosopis juliflora (PJ) was determined using acute oral toxicity tests using adult female Wistar albino rats.
Place and Duration of Study: Laboratories in the departments of Chemistry, Zoology, Botany and Biochemistry of Jomo Kenyatta University of Agriculture and Technology (J.K.U.A.T.) between March 2012 and April 2012.
Methodology: The OECD 425 guidelines (Up-and-Down procedure) were followed. Different dosages (control, 175, 550, 1750 and 5000 mg/kg body weight) were used in the experiment. Selective observations and analysis were made and recorded on mortality, signs of pain or distress and moribund animals, biochemical and macroscopic (pathological, organ and live body weights) analyses.
Results: During the entire period of the study, no signs of pain or enduring distress were observed neither was there any mortality. Alanine aminotransferase (ALT) values were within range (for experimental rats) apart from the rat in control while Aspartate aminotransferase (AST) values were within range (for experimental rat) apart from two rats in the upper limit. Macroscopic organ observations did not show colour or texture consistent with drug-induced inflammation or lesions. The toxicity studies of the extract mixture showed that the median lethal dose (LD50) was above the upper limit of 5000mg/kg body weight.
Conclusion: In conclusion, the LD50 of the extract mixture was found to be greater than 5000 mg/kg body weight and was, therefore, considered safe and has potential as a novel herbal preparation.

Open Access Original Research Article

Oral Administration of Euterpe oleracea Mart., Its Influence over Body Weight and Lymphoid Organs and Alterations in Haematological and Glycaemical Parameters

Mayara T. Pinheiro, Nayara S. R. Silva, Cleison C. Lobato, Francinaldo S. Braga, César F. Santos, Josivan S. Costa, José Carlos T. Carvalho, Cleydson Breno R. Santos

Journal of Pharmaceutical Research International, Page 2484-2493
DOI: 10.9734/BJPR/2014/10354

Aims: The aims of the study were to evaluate alterations caused by the oral administration of the aqueous extract of Euterpe oleracea to non-isogenic adult male mice during 15 and 30 consecutive days through the comparison of body and spleen weights, number of splenic and medullary cells, their hemogram and glycemia indices.
Study Design: Animals were divided into 8 groups (5 animals/group). Treated groups received the AEA concentration (125 mg/ml by gavage) at doses of 5, 50 and 500mg/kg for 15 and 30 days. The control group received only the vehicle of dilution (1x PBS) in volume of 0.5 ml. After treatment the animals were sacrificed in a CO2 chamber for testing.
Place and Duration of Study/Methodology: The aqueous extract of concentration of 125 mg/ml of lyophilized açaí pulp was prepared in Laboratory of General and Analytical Chemistry of Federal University of Amapá. Values of body weight, weight of liver and spleen, number of spleen and bone marrow cells, blood count and glucose of the mice were conducted on Drugs Research Laboratory between January and November of 2013.
Results: The statistical analysis was done by ANOVA test two-way with significance level p<0.05 in relation to control followed by Tukey post-test. The AEA caused significant changes in body weight, about 22% in animals treated with 500 mg/kg. Weight of spleen showed no significant changes, there was statistical difference in blood glucose levels between groups 5 and 500 mg/kg treated for 15 days and punctually in the 500 mg/kg group treated for 15 and 30 days. It was observed that the treatment with the AEA doses (5, 50 and 500 mg/kg) for 30 days increased the number of bone marrow cells. Regarding the number of spleen cells, treatment promoted changes, reducing the amount of cells during the 30 days of treatment, principally at the dose of 50 mg/kg. During the 15 day treatment of 500 mg/kg there was an increase in the number of spleen cells.
Conclusion: The treatment of mice with aqueous extract of açaí pointed that the concentration has significant influence in: 1) glucose concentration in the blood; 2) The increased number of bone marrow cells; 3) and the number of spleen cells. Thus, comparison between groups of animals was compatible with the hypothesis that the increase in body mass is a risk factor for insulin resistance.

Open Access Original Research Article

Extended Release Niosomal Hydrogel for Ocular Targeting of Piroxicam: In vitro and Ex vivo Evaluation

Bazigha K. Abdul Rasool, Oday S. Azeez, Hamda A. Lootah, Iman M. Abusharbain, Hiba A. Abu-Alhaj, Fazilatun Nessa

Journal of Pharmaceutical Research International, Page 2494-2510
DOI: 10.9734/BJPR/2014/13723

This study aimed at the investigation of piroxicam-niosomal hydrogel for ocular targeting to prolong and enhance its local analgesic activity. Various formulations were prepared, characterized and evaluated ex vivo for their transocular permeation using excised cow cornea. The prepared niosomes had distinct spherical multi-lamellar shape and mean vesicle size between 1.25±0.81μm and 7.47±0.85μm. Relevant increase in drug EE% was obtained with increase of cholesterol content and surfactant’s hydrophobicity. Drug retention in vesicles was significantly (p<0.05) higher at refrigerated condition than that at the room temperature. Prolonged drug release was achieved with high niosomal cholesterol content and the mechanism of drug release can be described as Fickian diffusion. The niosomal hydrogel showed 3.7 Permeability Improvement Ratio comparing to piroxicam aqueous suspension. The optimized niosomal gel showed prolonged drug release and enhanced piroxicam ocular bioavailability due to the formation of an amorphous drug form within the gel.

Open Access Original Research Article

Marine Streptomyces cyaneus Strain Alex-SK121 Mediated Eco-friendly Synthesis of Silver Nanoparticles Using Gamma Radiation

El-Batal A. I, Mohamed Helal El-Sayed, Bahgat Mohamed Refaat, Ahmed Abdel Zaher Askar

Journal of Pharmaceutical Research International, Page 2525-2547
DOI: 10.9734/BJPR/2014/12224

Aim: The present study aimed to develop cost-effective, eco-friendly marine Streptomyces cyaneus strain Alex-SK121 mediated synthesis of silver nanoparticles (AgNPs) with antimicrobial, antitumor and antioxidant activities.
Methodology: Aqueous 1mM silver nitrate (AgNO3) solution was treated with cell-free supernatant (CFS) of a novel Streptomyces cyaneus strain Alex-SK121 isolated from marine sediment samples. The prepared solution was irradiated with different doses of gamma rays ranged from 0.5 to 30.0kGy. Initial characterization of the synthesized AgNPs was performed by visual observation of color change in the prepared solution followed by analysis of UV-Visible Spectrophotometer (UV-Vis.), Fourier Transform Infrared Spectrometer (FT-IR), Dynamic Light Scattering (DLS) and Transmission Electron Microscopy (TEM). Evaluation of antimicrobial activity of the synthesized AgNPs against some pathogenic microorganisms was carried out. Antitumor activity of AgNPs was carried out against some human cancer cell lines using the method of Sulphorodamine B (SRB) assay, antioxidant activity of AgNPs was also studied using DPPH scavenging assay.
Results: In the present study, the cell-free supernatant of Streptomyces cyaneus strain Alex-SK121 isolated from sediment samples collected from Sidi Kerir region, Alexandria governorate, Egypt was found to reduce Agions to AgNPs. Identification of the producer strain was performed according to spore morphology and cell wall chemo-type, which suggested that this strain is Streptomyces. Further cultural, physiological characteristics and analysis of the nucleotide sequence of 16S rRNA gene indicated that this strain is identical to Streptomyces cyaneus and then designated Streptomyces cyaneus strain Alex-SK121. To maximize the production of AgNPs, the tested supernatant was irradiated with different doses of gamma rays and it was found that, 15 kGy is the best applied dose induces AgNPs synthesis. The synthesized AgNPs showed the characteristic absorption spectra in UV–Vis. at 425 nm. The microbiologically synthesized AgNPs showed significant antimicrobial activity towards some pathogenic microorganisms with inhibition zone ranged from 13 up to 20 mm. Also AgNPs exhibited antitumor activity against human breast carcinoma cells and human liver carcinoma cells with IC50 9.63 and 33.75 µg/ml respectively in addition to 96% antioxidant activity.
Conclusion: Gamma irradiation which induced AgNPs synthesis by cell-free supernatant of marine actinomycetes Streptomyces cyaneus strain Alex-SK121 with different applications is a simple, clean, economic and environmental friendly approach.

Open Access Original Research Article

Evaluation of the Quality of Commercial Antibacterial Discs Available in Nigeria

P. M. Eze, E. E. Ajaegbu, P. C. Ejikeugwu, R. N. Egbuna, C. C. Abba, C. O. Esimone

Journal of Pharmaceutical Research International, Page 2548-2562
DOI: 10.9734/BJPR/2014/10900

Aims: This study was undertaken to compare the quality and efficacy of six brands of antibacterial discs that are commercially available in Nigeria.
Methodology: The brands evaluated include two foreign brands (Oxoid and Abtek) and four local brands (Optudisc, Polydisc, Maxidisc and Jirehdisk). The brands were analyzed by antibiotic susceptibility testing (AST) using laboratory isolates of Staphylococcus aureus and Escherichia coli to measure the antimicrobial performances of the brands; and UV-Vis spectrophotometry to measure the absorbances of antibiotics extracted from antibiotic discs of the various brands.
Results: All of the brands of antibacterial discs of under study exhibited variations in their antimicrobial performances and UV-absorbances. This was observed where some of the discs with lower stated potencies produced inhibition zones and absorbances far greater than similar discs from other brands with higher stated potencies. Also, discs of the same stated potencies showed variable results in both the antibiotic susceptibility testing (AST) and UV-Vis spectrophotometric analyses. Coefficients of variation greater than 5%, which indicates high disc-to-disc variation and unsatisfactory reproducibility, were recorded highest among the local brands during the AST. All the brands with multidisc panels, except the Abtek and Polydisc brands, produced some zones of inhibition that are unreadable. Of all the zones of inhibition that were unreadable, Optudisc brand recorded the highest rate (36∙7%), while 6∙7% of discs of Jirehdisk brand and 6∙7% of discs of Maxidisc brand produced inhibition zones that were unreadable.
Conclusion: All brands of susceptibility discs evaluated in this study except the Oxoid and Abtek brands manifested poor quality and performed below expected standard, though one of the local brands (Polydisc) performed closest to the foreign brands. With further improvement in quality, these brands may be recommended for use in Nigeria.

Open Access Original Research Article

Characterization of Bioactive Components of Gossypium barbadense L. with Hematinic Potential in Wister Albino Rats

Zara Muhammad, Samuel Atabo, Amina Zakari

Journal of Pharmaceutical Research International, Page 2563-2574
DOI: 10.9734/BJPR/2014/13194

Aims: To ascertain the hematinic potential and bioactive compounds in Gossypium barbadense.
Place and Duration of Study: Department of Pharmaceutical Sciences, Ahmadu Bello University, Zaria, and Department of Applied Science, College of Science and Technology, Kaduna Polytechnic, Kaduna, Nigeria between February, 2013 and July, 2014.
Methodology: Forty eight (48) apparently healthy albino rats weighing (150-200g) were grouped in to seven groups of five rats each. Thirteen rats were used for the G. barbadense toxicity test. Hemolytic anemia was induced using Phenylhydrazine (10mg/kg bw). Different doses (100mg/ml, 200mg/ml, and 400mg/ml) of G. barbadense were administered with periodic evaluation of Haematological indices (Hemoglobin concentration, Packed Cell Volume, Red Blood Cells and reticulocyte count). Bioferon (0.23ml/kg b.w) was used as the standard drug. Synergistic Thin layer Chromatography and Column Chromatography were used to purify the plant extract. Gas Chromatography linked with Mass Spectroscopy (GC-MS) was used in the Characterization of purified fraction.
Results: The level of Hb (g/dl) was found to increase in a dose dependent manner (100mg-12.17g/dl, 200mg-12.60g/dl and 400mg-12.87 g/dl), likewise RBC (4.31, 4.41 and 4.72) and PCV (43.35%, 43.49%, 43.65%). Characterization revealed the presence of 19 compounds.
Conclusion: G. barbadense resulted in HB, RBC and PCV boost, owing to inherent bioactive component.

Open Access Review Article

Limitations of Conventional Parameters and Role of Urinary Protein Biomarkers in the Determination of Drug Induced Acute Kidney Injury in Very Early Stage

R. Shanmuga Sundaram, B. R. Abhirama, L. Gowtham, Gladys Kalpana, M. Sudha, T. Thiyagarajan, S. Pushpa, G. K. Priyadharsini

Journal of Pharmaceutical Research International, Page 2463-2474
DOI: 10.9734/BJPR/2014/13446

Drug-induced Acute Kidney Injury (AKI) constitutes an important cause of acute renal failure and chronic kidney disease in present day clinical practice. Drug-induced acute renal failure (ARF) accounted for 20% of all ARF in an Indian study. The incidence and prevalence of chronic kidney disease (CKD) has dramatically increasing worldwide. Progression of AKI from mild or moderate to end stage may be prevented by selecting potentially effective therapies, if it is detected in very early stage. But early detection of AKI is often difficult due to paucity of early predictive noninvasive biomarkers. Development of omics technology has led to the identification of several urinary protein biomarkers and transcriptional biomarkers, which enable earlier detection of kidney injury. Urinary protein biomarkers have great benefit due to the easy or non-invasive availability of urine and many showing good predictive power. Several urinary protein biomarkers have been identified and have demonstrated superiority in detecting kidney injury in comparison to conventional parameters like serum creatinine (SCr), blood urea nitrogen (BUN) etc. These promising experimental biomarker of kidney damage require further confirmation of its use in routine clinical use.