IN VIVO CHARACTERIZATION OF LESS PAINFUL PROPOFOL NANOEMULSION USING PALM OIL FOR INTRAVENOUS DRUG DELIVERY

Objective: The objective of present work was to evaluate the effectiveness of propofol in nanoemulsion based palm oil that called as NEMS™, which was a choice of anesthetic drug to induce and maintenance general anesthesia to reduce pain on injection activity and also to evaluate the in vivo characterization of propofol in NEMS™. Methods: Preparation of propofol nanoemulsion using NEMS™ technology has been performed for propofol 1% in NEMS™ (P1%), and propofol 2% in NEMS™ (P2%). Determination of free propofol concentration in aqueous phase was conducted using HPLC and rat paw lick test was evaluated as in vivo test to assay the intensity of pain on injection site. The sleep recovery test was conducted to evaluate the pharmacological effect and erythrocyte hemolysis test also conducted to ensure the safety of propofol in NEMS™. All of the test results were compared with Diprivan®1% as a positive standard. Results: The contents of free propofol in formulation P1% and Diprivan®1% in aqueous-phase were 6.20±0.03 µg/ml and 15.02±0.33 µg/ml, respectively (*P<0.05). The rat paw lick test showed that the formulation P1% was significantly (*P<0.05) less painful when compared to Diprivan®1%. There were no significant differences in pharmacological effect for all of the formulations (*P>0.05). The erythrocyte haemolysis test show that all formulation still safe for our blood. Conclusion: Palm oil can be used as a carrier for propofol and it was successfully reduced the free propofol contents and the intensity of pain on injection site in rats

Interaction between green tea and perindopril reduces inhibition of angiotensin-converting enzyme activity

Purpose: To investigate the blood pressure-lowering effect of green tea (GT) extract alone and in combination with an angiotensin converting enzyme (ACE) inhibitor, perindopril, on rats. Methods: The study consisted of four groups of five spontaneously hypertensive rats (SHR): negative control (2 % tragacanth mucilage), positive control group (perindopril, 0.36 mg/kg/day) and two treatment groups (green tea, 25 mg/kg/day; and combined green tea/perindopril). The treatments were given orally for 14 days. Systolic blood pressure was measured before and after treatment using the tail cuff technique. Angiotensin converting enzyme activity in the lung homogenate of the hypertensive rats was determined spectrophotometrically. Results: Green tea extract significantly reduced the rats’ systolic blood pressure (p < 0.05) but did not inhibit the angiotensin-converting enzyme. The combination of green tea extract with perindopril also caused a significant decline in blood pressure (p < 0.001). However, the green tea extract attenuated the inhibition of the angiotensin-converting enzyme activity by perindopril. Conclusion: Green tea extract produces anti-hypertensive activity in rats, but its mechanism of action is not via inhibition of angiotensin-converting enzyme. The interaction of GT extract with perindopril results in a reduction of ACE inhibitory activity

Crossover learning through a health campaign integrated in a bachelor of pharmacy curriculum

Purpose: Continuous improvement in teaching and learning methods is vital for addressing the changing needs of pharmacy education. Many have agreed that one of the most effective approaches is crossover learning, whereby learners are actively involved in shaping their learning experiences and acquiring knowledge in both formal and informal settings. In this work, we report an ongoing initiative of facilitating a student-led health campaign, Brain Awareness Day, to promote crossover learning. Method: The campaign is included in our curriculum (Bachelor of Pharmacy with Honours) as a formal mode of learning and continuous assessment. A total of 84 pharmacy students were divided into groups of eight or nine to work on a drug addiction-themed assignment and present the result in the form of a poster exhibition. A short, online questionnaire was used to gather the students’ feedback on their learning experience and perceived gain of relevant insights from the campaign. Findings: Thirty nine out of 84 students took part in the survey. Most students agreed that their involvement in the campaign had contributed favourably to their learning experience and achievement of the pre-defined learning outcomes. The students also gave several suggestions for improving the organisation of the campaign. They suggested that more budget should be allocated for running the campaign, and that finding an off-campus venue might help to increase footfall. Significance: We concluded that the campaign had been effective in encouraging crossover learning, and it would remain an integrated sub-programme in our pharmacy curriculum. Diversifying methods of teaching and learning may help to realise Malaysia’s aim of developing well-rounded and employable graduates

Effect of Allium sativum (garlic) methanol extract on viability and apoptosis of human leukemic cell lines

Purpose: To investigate the effect of Allium sativum (garlic) methanol extract on viability and apoptosis of human leukemic cells.Methods: Cell viability was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay at concentrations of 3.125, 6.25, 12.5, 25, 50, 100, 200, 400 and 800 ug/mL of Allium sativum extract following 48-h treatment on U-937, Jurkat Clone E6-1 and K-562 cell lines. The mode of cell death was determined by Annexin V-FITC staining and analyzed by flow cytometry.Results: The results show that the half-maximal inhibitory concentration (IC50) of A. sativum on U-937, Jurkat Clone E6-1, K-562 cell lines was 105 ± 2.21, 489 ± 4.51 and 455 ± 3.13 μg/mL, respectively, compared with negative control, while apoptosis was 17.93 ± 0.95 % for U-937 cells (p ≤ 0.05), 38.37 ± 1.88 % for Jurkat Clone E6-1 cells (p ≤ 0.001) and 16.37 ± 1.10 % for K-562 cells. A majority of the cells were inhibited by the extract via apoptosis. Only U-937 cells (6.87 ± 0.65 %) showed significant necrosis compared to negative control (p ≤ 0.05).Conclusion: K-562 cells are the most resistant against garlic extract, in contrast to Jurkat Clone E6-1 cells. Garlic extract does not induce necrosis in Jurkat Clone E6-1 and K-562 cells.Keywords: Anti-leukemic, Garlic, Allium sativum, Annexin V-FITC staining,  Necrosis, Apoptosis, Flow cytometry, Jurkat Clone E6-1 cell

A brief review of potential neuroprotective roles of the culinary herb Ocimum basilicum

Neurodegenerative diseases commonly affect elderly population and are characterised by progressive neuronal loss. Oxidative stress is highly associated with neurodegeneration. The targeted herbal plant in this review, Ocimum basilicum (O. basilicum), is typically used in Indochina and Italian cuisine. Pharmacological studies on O. basilicum have demonstrated potent antioxidant activities with some reports of neuroprotective actions. This brief review highlights the potential neuroprotective roles of O. basilicum by discussing previously documented antioxidative actions of the plant extract, essential oils and its phytochemical compounds on the nervous system based on in vitro and in vivo studies. Accumulating evidence on the neuroprotective action of O. basilicum points to a notion that neuroprotection is made possible by way of its antioxidant properties and largely due to the presence of polyphenol compounds such as rosmarinic acid which has been identified as the major constituent. Although the mechanisms of O. basilicum antioxidant action have been proposed, further studies are required for better understanding of its antioxidant action leading to neuroprotective roles. It is also possible that the antioxidant actions of O. basilicum are mediated through synergism of a mixture of various naturally-occurring bioactive compounds in the plant, as is with many other plant-based food supplements, to produce the putative effects instead of a single bioactive compound from the plant. Therefore, specific targeting of neuroprotection by means of antioxidant actions warrants further preclinical and clinical studies investigating the therapeutic potentials of O. basilicum particularly in view of the prevention of neurodegenerative processes

Effects of durian fruit on blood pressure of spontaneously hypertensive rats

Durian or scientifically known as Durio zibethinus is one of the most well-known seasonal fruits in the Southeast Asia region. However, its safe consumption in individuals with hypertension is still controversial. This study was conducted to investigate the effect of durian on blood pressure of spontaneously hypertensive rat model. Four groups of rats (n=5) were fed with either a low dose durian (26 g/kg), a high dose durian (52 g/kg), sugar solution (8 mL/kg) which has similar sugar composition in the durian as placebo control, and distilled water as vehicle control (8 mL/kg) for 14 days. The durian doses for rats were obtained by converting from human doses. Baseline reading of blood pressure and heart rate were recorded before the first oral administration of durian. The blood pressure and heart rate were also measured 1 h after the durian oral administration on day 1, 3, 7 and 14 of the experiment. In conclusion, durian fruit possessed an acute effect on the blood pressure of hypertensive rats but heart rate was unaffected. High dose administration of durian led to significant elevation of blood pressure after 1 h of consumption. Meanwhile, low dose of durian (26 g/kg) caused an insignificant reduction in systolic and diastolic blood pressure. Tolerance to the durian fruit was observed after three to seven days of the oral administration and low dose consumption of durian fruit was safe in the hypertensive rat

Acute oral toxicity and biodistribution study of zinc aluminium-levodopa nanocomposite

Layered double hydroxide (LDH) is an inorganic-organic nano-layered material that harbours drug between its two-layered sheets, forming a sandwich-like structure. It is attracting a great deal of attention as an alternative drug delivery (nanodelivery) system in the field of pharmacology due to their relative low toxic potential. The production of these nanodelivery systems, aimed at improving human health through decrease toxicity, targeted delivery of the active compound to areas of interest with sustained release ability. In this study, we administered zinc-aluminium-LDH-levodopa nanocomposite (ZAL) and zinc-aluminium nanocomposite (ZA) to Sprague Dawley rats to evaluate for acute oral toxicity following OECD guidelines. The oral administration of ZAL and ZA at a limit dose of 2,000 mg/kg produced neither mortality nor acute toxic signs throughout 14 days of the observation. The percentage of body weight gain of the animals showed no significant difference between control and treatment groups. Animal from the two treated groups gained weight continuously over the study period, which was shown to be significantly higher than the weight at the beginning of the study (P < 0.05). Biochemical analysis of animal serum showed no significant difference between rats treated with ZAL, ZA and controls. There was no gross lesion or histopathological changes observed in vital organs of the rats. The results suggested that ZAL and ZA at 2,000 mg/kg body weight in rats do not induce acute toxicity in the animals. Elemental analysis of tissues of treated animals demonstrated the wider distribution of the nanocomposite including the brain. In summary, findings of acute toxicity tests in this study suggest that zinc-aluminium nanocomposite intercalated with and the un-intercalated were safe when administered orally in animal models for short periods of time. It also highlighted the potential distribution ability of Tween-80 coated nanocomposite after oral administration

Quantification of BCR-ABL transcripts in peripheral blood cells and plasma of chronic myeloid leukemia patients at different stages of tyrosine kinase inhibitor treatment response

Purpose: To investigate the feasibility of using peripheral blood plasma samples as surrogates for blood cell sampling for quantification of breakpoint cluster region-Abelson oncogene (BCR-ABL) transcript levels to monitor treatment responses in chronic myeloid leukemia (CML) patients.Methods: Peripheral blood samples were collected from 20 healthy individuals and 165 CML patients at various stages of treatment response. Level of BCR-ABL mutation gene transcripts were determined by RNA extraction and quantitative real time polymerase chain reaction (RT-PCR).Results: The ratio of BCR-ABL transcripts/ABL transcripts in blood cells was significantly higher (p &lt; 0.05) than in plasma for untreated and treated patients. Patients who received tyrosine kinase inhibitor (TKI) therapy and achieved major molecular response during treatment had the lowest mean ratio detected in blood cells (0.06 ± 0.0 %) and the ratio was below the detectable limit in plasma samples. Unlike plasma samples, BCR-ABL/ABL transcript ratios in blood cell samples strongly correlated with white cell counts in CML patients undergoing all types of treatment responses.Conclusion: Blood cell sampling is more sensitive than plasma sampling in diagnosis and evaluation of CML treatment response.Keywords: Chronic myeloid leukemia, Peripheral blood cells, Plasma RNA,  Treatment response, Tyrosine kinase inhibitor

Neuroprotective effects of Ocimum basilicum extract against hydrogen peroxide-induced oxidative stress in SK-N-SH neuroblastoma cells

Neurodegenerative diseases such as Alzheimer’s and Parkinson’s disease are characterized by the progressive loss of neurons. One of the contributing factors for these diseases is oxidative stress, characterized by the imbalance of free radicals production and antioxidant defense mechanisms. In the present study, the neuroprotective effects of Ocimum basilicum var. thyrsiflora against hydrogen peroxide (H2O2)-induced oxidative stress in SK-N-SH neuroblastoma cells were evaluated. The exposure of SK-N-SH cells to 50 μM H2O2 for 24 h induced cytotoxicity and apoptosis as measured by cell viability and flow cytometry, respectively. Pretreatment with ethyl acetate (ObEA) fraction at 3.1-25 μg/mL showed the highest protection against H2O2-induced cell death compared to other fractions and crude extract by increasing cell viability and reducing apoptosis. The evaluation of antioxidant capacity via 1, 1-diphenyl-2-picrylahydrazyl (DPPH) and ferric reducing/antioxidant power (FRAP) assays showed ObEA possessed the highest antioxidative properties. The intracellular reactive oxygen species (ROS) production of H2O2 in untreated cells increased by 2.39-fold compared to the control and was significantly attenuated by the 2 h pre-treatment of O. basilicum (p<0.05). The reduction in intracellular superoxide dismutase (SOD) induced by H2O2 was also abrogated by the pretreatment of O. basilicum. These findings suggested that O. basilicum is potentially neuroprotective against oxidative damage in neuronal cells by scavenging free radicals, restoring SOD activities and eventually prevent cell death

P2Y purinergic receptor signaling in oral squamous cell carcinoma cell lines and its role in proliferation and cisplatin-mediated apoptosis

Treatment of advanced stage oral squamous cell carcinoma (OSCC) often involves the use of chemotherapeutic agents, such as cisplatin. However, its use often results in therapeutic failure due to chemoresistance. This study focused on a class of purinergic receptors, namely P2Y, which are activated via interaction with extracellular nucleotides. The functional effects of P2Y receptor activation in OSCC cell lines as well as the signaling pathways involved were investigated. The expression of P2Y2 receptors in histological sections of OSCC was studied due to its association with cancer. Activation of MAPK pathways via extracellular nucleotides were studied in OSCC cell lines, along with downstream effects such as proliferation and cisplatin-mediated apoptosis. Immunohistochemical staining of OSCC tissue samples showed loss of P2Y2 expression as the disease progressed. Western blotting identified different MAPK signaling pathways were activated by extracellular nucleotides. Bromodeoxyuridine proliferation assays showed increased cellular proliferation in the OSCC cell lines H400 (p < 0.001) and SAS (p < 0.001) after 24 h treatment with ATP. However, the ability of extracellular nucleotides to activate multiple P2Y receptor subtypes may indicate the involvement of other subtypes aside from P2Y2. Cisplatin-mediated apoptosis was enhanced in SAS cells co-treated with ATP (p < 0.001), while H376 (p < 0.001) showed reduction in the number of apoptotic cells and no significant changes were observed in H103. This study concluded that extracellular nucleotide on OSCC cell lines with different characterizations had varied downstream effects, which suggests the use of targeted therapy to specific individuals