Open Access Original Research Article

Japanese Kampo Medicine, Ninjinyoeito, Inhibits the Induction of iNOS Gene Expression in Proinflammatory Cytokine-Stimulated Hepatocytes

Yoshito Tanaka, Masaki Kaibori, Hirokazu Miki, Masaharu Oishi, Richi Nakatake, Katsuji Tokuhara, Mikio Nishizawa, Tadayoshi Okumura, A-Hon Kwon

Journal of Pharmaceutical Research International, Page 2226-2244
DOI: 10.9734/BJPR/2014/13301

Background: Japanese Kampo medicine, ninjinyoeito (NYT), is used for the treatment of complaints in patients with general fatigue, anorexia and anemia, and is recently applied in patients with chronic hepatitis C. However, there is little scientific evidence to demonstrate the liver-protective effects of NYT. We examined proinflammatory cytokine stimulated hepatocytes as a simple in vitro liver injury model to determine liver-protective effects of NYT and to clarify the mechanisms involved in.
Methods: Primary cultured rat hepatocytes were treated with interleukin (IL)-1β in the presence or absence of NYT. Inflammatory biomarkers including inducible nitric oxide synthase (iNOS) and tumor necrosis factor (TNF)-α were analyzed.
Results: IL-1β stimulated the induction of iNOS and enhanced the production of NO. Simultaneous addition of NYT decreased the levels of iNOS protein and its mRNA expression, resulting in the reduction of NO production. NYT also reduced the expression of TNF-α mRNA and enhanced the expression of IL-10 mRNA. NYT inhibited two essential signaling pathways for iNOS induction, the activation of nuclear factor (NF)-κB and the upregulation of type I IL-1 receptor. Experiments with transfection and iNOS gene antisense transcript revealed that NYT reduced the levels of iNOS mRNA at both the promoter activation and mRNA stabilization steps. The delayed administration of NYT after IL-1β addition also inhibited iNOS induction.
Conclusions: Results demonstrate that NYT can influence the induction of inflammatory mediators, such as iNOS and TNF-α, in part through the inhibition of NF-κB activation in hepatocytes. NYT may have therapeutic potential for various acute liver injuries.

Open Access Original Research Article

Use of Mobile Phones for Monitoring Adverse Drug Reaction in Pharmacy and Drug Stores in Ishaka, Uganda - a Pilot Assessment of Willingness to Report

A. A. Adedeji, J. Babirye, O. Nsooli, D. Kamowa, O. A. Tikare, A. G. Okoruwa, J. O. C. Ezeonwumelu, A. A. Ganiyu, C. P. Adiukwu, J. Namulema, C. Barigye, S. Yusuf, M. Vicente-Crespo

Journal of Pharmaceutical Research International, Page 2245-2260
DOI: 10.9734/BJPR/2014/8065

Aim: We investigated reporting of Adverse Drug Reaction (ADR) following use of drugs purchased from open system pharmacy (OSP) and drug stores, and the effectiveness of mobile phones for reporting drug reactions and detection of drug interactions.
Study Design: The study was descriptive and inceptional.
Place and Duration of Study: Selected Pharmacies and drug stores in Ishaka Municipality, Bushenyi, Uganda, between January and April 2012.
Methodology: A total of 190 participants purchasing prescription and non prescription drugs in the drug outlets were enrolled and drug purchases documented. Structured interviews were used to assess any existing system of ADR tracking. Possible interactions were assessed using electronic checkers software on drug combinations prescribed or purchased. Mobile phone calls were used to monitor the reporting potential, use of medication and events or reactions following drug use for ADRs.
Results: No formalized pre-study system was found for tracking ADR in the OSP and drug stores studied. Participants purchased 420 different medications with 55.8% without prescription. Antibiotics, analgesics and antimalarials ranked most purchased medications. All participants carried at least a functional mobile phone and demonstrated interest to report ADRs. Mean Effective Mobile Phone Contact Ratio (MEMPCR) for ADR monitoring was 0.91+0.2 and follow-up was 96% (n=183) and 89.5% on days 0 and 4 respectively. Interactions predicted were in 24.8% (31). Significant reporting of at least one of 404 reactions occurred within 72hr compared to 96-120hr (P=0.003). Two participants had reaction leading to discontinued use of Cotrimoxazole.
Conclusion: Use of mobile phones and drug interaction checker software may avail early detection of ADR and reporting. Facilitated toll free- call service may be an effective means of extending the scope of ADR tracking in addition to Yellow Card scheme, and augment involvement of pharmacists and consumers in safe use of drugs.

Open Access Original Research Article

Design, Formulation and Tableting Properties of Aqueous Leaf Extract of Moringa oleifera

J. Muazu, Z. A. Suleiman

Journal of Pharmaceutical Research International, Page 2261-2272
DOI: 10.9734/BJPR/2014/11516

Aim: The aim of this study is to formulate a standard dose of aqueous extract of Moringa oleifera leaves into tablets and to determine a suitable binder for the formulation.

Methodology: Aqueous extract of Moringa oleifera leaves was extracted and formulated using different binders which included Maize Starch, Gelatin and Micro-crystalline Cellulose (MCC) to find out which one produce better tablets of aqueous extract of Moringa oleifera leaves. Formulations were characterized using various parameters such as physicochemical properties (bulk density, tapped density, moisture content, Hausner’s ratio, Carr’s index, ash value), strength (friability and crushing strength) and release properties (disintegration and dissolution times tests). The result showed that tablets formulated with Gelatin as a binder has lowest friability and disintegration time compared to those formulated with either MCC or maize starch. The crushing strengths were all within the acceptable limit (3 – 6 KgF) except maize starch which was higher.
Conclusion: Moringa oleifera tablets were successfully formulated and based on experiments conducted, Gelatin is preferable in the formulation of Moringa oleifera tablets.

Open Access Original Research Article

Effects of Wild Artemisia herba-alba Essential Oil on Biofilm-Forming Bacteria

Jehad Al-Shuneigat, Sameeh Al-Sarayreh, Yousef Al–Saraireh, Ibrahim Al-Tarawneh, Mahmoud Al-Qudah, Eman Albataineh

Journal of Pharmaceutical Research International, Page 2273-2280
DOI: 10.9734/BJPR/2014/12605

Aims: The aim of the present study was to determine the effects of essential oil of wild Artemisia herba-alba grown in south Jordan on biofilm-forming bacteria.
Study Design: Seven bacterial clinical isolates were used in this study. Biofilm formation was first quantified in microtitre plates. The minimum inhibitory concentration (MIC) and biofilm inhibitory concentration (BIC) assays were performed in microtitre plates using a two fold dilution series. The most tolerant isolate were then used to test the effectiveness of Artemisia herba-alba essential oil on initial adherence to polystyrene surface.
Place and Duration of Study: Faculty of Medicine, Mutah University, Mutah, Jordan; Faculty of Science, Department of Chemistry Yarmouk University, Irbid, Jordan; and Faculty of Science, Department of Chemistry, Albalqa' Applied university, Albalqa, Al-Salt, Jordan. Between August 2012 and June 2014.
Methodology: Using a microtitre plate assay we measured inhibitory adherence effect for Artemisia herba-alba essential oil against seven biofilm-forming bacterial clinical isolates.
Results: Artemisia herba-alba essential oil produced inhibitory effects against all isolates and susceptibility varied considerably. The MIC values were found to be in the range of 0.5-4% v/v. In addition Artemisia herba-alba essential oil was able to inhibit initial adherence in the most tolerant isolate (Pseudomonas aeruginosa) at sub-inhibitory concentrations.
Conclusions: Artemisia herba-alba essential oil showed a significant activity against all isolates. It was able to inhibit initial adherence in the most tolerant isolate at sub-inhibitory concentrations.

Open Access Original Research Article

Formulation and Evaluation of In Situ forming Polymeric Drug Delivery Systems for Mixed Vaginal Infection

Haidy Atef Abass, Rabab Kamel

Journal of Pharmaceutical Research International, Page 2281-2295
DOI: 10.9734/BJPR/2014/12089

Aims: Vaginal preparations are still associated with number of problems including frequent administration and escape from vagina causing discomfort to patient. For efficient vaginal delivery of drugs, the delivery system should reside at the site of infection for a prolonged period of time therefore this work aims to prepare Vaginal Capsules containing sustained release in situ forming polymeric particles containing broad spectrum antibiotics to cover all the common pathogen associated with vaginal infections.
Study Design: Characterization for the developed beads, such as determination od, particle size, drug entrapment yield, and drug release profiles, were characterized prior to determining intracellular uptake profile, in vitro, and in vivo tissue distribution patterns of the particles.
Place and Duration of Study: Department of pharmaceutical technology in German university in Cairo and Department of Pharmaceutical Technology in National Research Center, Cairo, Egypt between June 2012 and March 2014.
Methodology: Calcium alginate, chitosan and mixed polycarbophil beads containing fluconazole (antifungal) and metronidazole (antiprotozoal) were packed in hard gelatin capsule and evaluated as new vaginal drug delivery forms beads were characterized by size, Scanning Electron Microscopy (SEM), weight uniformity and drug entrapment efficiency as well as in –vivo bioadhesion test
Results: Results of the in vitro antimicrobial study indicated that the M5 and F4 mixed beads had better antimicrobial action than the commercial intravaginal drug delivery systems and retention was prolonged in an ex vivo retention study showed that the bioadhesion of the beads were 68.88 to 84.3%.
Conclusion: Developed beads were found to be more effective in in vitro conditions. The average zone of inhibition of the developed M5 the formulae that contain (Metronidazole: Chitosan: Polycarbophil) in (1:1:1 ratio)and F4the formulae that contain (Fluconazole: Chitosan: Polycarbophil) in (1:1:1 ratio) mixed beads Candida albicans was 28.3±0.6 mm compared to 24.5±1.7 mm of commercial Amrizol®, indicating significantly higher efficacy of M5 and F4 mixed beads (p>0.001). Average zone of inhibition of the developed M5 and F4 mixed beads against E. coli was 31.5±2.2 mm compared to 28.6±1.8 mm of Amrizol®. The developed M5 and F4 mixed beads was more effective than the tested commercial formulations and last for12, 18 and 24 hr.

Open Access Original Research Article

Development of Ultrasound Sensitive eLiposomes Containing Doxorubicin for Drug Delivery

Chung-Yin Lin, William G. Pitt, Kuo-Chen Wei

Journal of Pharmaceutical Research International, Page 2296-2311
DOI: 10.9734/BJPR/2014/13143

Aims: A novel nanocarrier was formulated by remote loading of doxorubicin (Dox) into a dipalmitoylphosphatidylcholine (DPPC) liposome that also contains various perfluorocarbon (PFC) droplets within its aqueous interior. It was shown that Dox can be loaded to a level of up to 67% into these large unilamellar vesicles composed of DPPC and cholesterol by employing a transmembrane pH gradient technique.
Methods: The different encapsulation efficiencies for these eLipoDox constructs of differing PFC composition are 45.5% (PFC5), 31.5% (PFC6) and 66.7% (PFC5/PFC6 mixture, PFCm). At 30 seconds of insonation, the eLipoDox formulation with PFCm droplets appeared to release more Dox than did eLipoDox with pure PFC5 or PFC6 droplets. The thermal stability of these eLipoDox formulations were examined at 37ºC at different times; then controlled delivery was demonstrated by applying low-frequency ultrasound (US) at 1 W/cm2.
Results: The eLipoDox with PFC6 or PFCm showed the best combination of thermal stability and drug release. An immunoblotting analysis indicates that ultrasound-triggered Dox release from eLipoDox could provide a higher quantity of nanodrug into tumor cells and thus may have cytostatic effects in cancer cells.
Conclusion: These eLipoDox constructs with low boiling point PFCs have the potential to provide more effective ultrasonically activated drug therapy to a desired location.

Open Access Original Research Article

A New Bioanalytical Method Development & Validation for Simultaneous Estimation of Esomeprazole and Naproxen in Human Plasma by Using RP-HPLC

S. Ashutosh Kumar, Manidipa Debnath, J. V. L. N. Seshagiri Rao, D. Gowri Sankar

Journal of Pharmaceutical Research International, Page 2312-2327
DOI: 10.9734/BJPR/2014/10918

Aim: A new reverse phase high performance liquid chromatography (RP-HPLC) method for the quantitative determination of Esomeprazole and Naproxen in human plasma was developed and validated as per US-FDA guidelines.
Methodology: The drug was spiked in the plasma and extracted with mobile phase by precipitation method. The extracted analyte was injected into Symmetry C18 (4.6 x 150mm, 5μm, Make: XTerra) or equivalent, maintained at ambient temperature and effluent was monitored at 285nm. The mobile phase was composed of potassium dihydrogen phosphate and acetonitrile [HPLC Grade] in the ratio of 60:40. The pH of the potassium buffer was adjusted to 3.0 by using Ortho Phosphoric Acid. The flow rate was maintained at 1.0 mL/min.
Results: The developed method shows high specificity for Esomeprazole and Naproxen. The calibration curve for Esomeprazole and Naproxen was linear from 1.0 to 6.0 ppm (r2= 0.999) and 25.0 to 150.0 ppm (r2= 0.999) respectively. The inter-day and intra-day precision was found to be within limits. The proposed method was adequate sensitivity, reproducibility, and specificity for the determination of esomeprazole and naproxen in plasma. The Lower limit of quantification (LLOQ) for the drug Esomeprazole and Naproxen were found to be 0.04µg/ml and 0.4µg/ml respectively. The average percent recovery for the drugs Esomeprazole and Naproxen were found to be 98.97-99.84 & 99.80-100.95 respectively and reproducibility was found to be satisfactory.
Conclusion: The proposed method was accurate, and precise for the quantification of Esomeprazole and Naproxen in the plasma. The proposed can also be used for routine analysis in quality control. The method was validated for parameters like selectivity, sensitivity, precision, intermediate precision, accuracy, linearity, recovery & stability. This RP -HPLC method is suitable for determining the concentration of Esomeprazole and Naproxen in plasma and it can applied for routine analysis for determination of the Esomeprazole and Naproxen from dosage form during pharmacokinetic study.

Open Access Original Research Article

Adherence to Anti-retroviral Therapy among HIV-Positive Individuals in Ghana: The Role of Stress and Post Traumatic Stress Symptoms

Irene A. Kretchy, Barima A. Afrane, Emmanuel Asampong, Cynthia E. Sena, Franklin Acheampong, Benedict Mbeah-Baiden

Journal of Pharmaceutical Research International, Page 2328-2339
DOI: 10.9734/BJPR/2014/13278

Aims: To identify stress and post traumatic stress symptoms (PTSS) in HIV patients on Antiretroviral therapy (ART) and further assess the possible relationship between stress, PTSS, immune function and ART medication adherence.

Methods: After obtaining written informed consent, 150 HIV out-patients attending Pantang and Amasaman Hospitals in Accra, Ghana, were recruited for the study. Participants were interviewed using a questionnaire comprising of sections measuring demographic characteristics, perceived stress, post traumatic stress symptoms, and medication adherence. A measure of the immune function (CD4+) from individual’s health records was noted at the time of interview.

Results: Ninety four (62.7%) patients exhibited higher stress levels compared to the remaining 56(37.3%). The majority of participants (74%) showed PTSS at levels that met the optimal criteria indicative of post traumatic stress disorder (PTSD) and may have required clinical attention and management.  Participants with high stress levels were 84% less likely to be adherent to their antiretroviral medication compared to those with lower levels of stress [OR=0.16 (0.050–0.512), P<0.0001]. Patients who showed evidence of stress were approximately four times more likely to develop PTSD compared to non-stressed patients [OR=3.898 (1.428–10.635), P=0.007]. The experience of side effects did not influence adherence [OR = 0.536 (0.224–1.285), P = 0.155]. On the other hand, stress significantly related with adherence when side effects were present [OR=0.648 (0.252– 1.662), P=0.002]. The PTSD importantly associated with adherence to ART with approximately 42% less odds of attaining optimal adherence by participants with PTSD compared to non-PTSD patients [OR=0.576 (0.189–1.751), P=0.017].

Conclusion: HIV patients on ART need to be screened for stress, PTSS and PTSD in order to benefit from psychological support and therapy. This activity can significantly enhance medication adherence as well as impact positively on the general health outcomes of HIV patients on ART.