Highly Efficient Activated Carbon (HEAC) as an adsorbent of toxins has been successfully produced from palm shell through chemical activation process using phosphoric acid as an activating agent. Palm Kernel shell used as the main raw material for activated carbon production, was purchased from a local oil palm mill in Pahang, Malaysia. Temperature range 550 ºC – 650 ºC was used during the activation process. The effect of temperature variation on the pore size and surface morphology of the activated carbon were studied. Well developed pore size and low number of functional groups observed on activated carbon at 600 ºC were determined by Scanning Electron Microscope (SEM) and Fourier- Transform Infrared (FTIR) spectroscopy, respectively. The surface area and pore volume were determined by Brunauer, Emmett and Teller (BET) method using N2 gas adsorption. The highest surface area (1287 m2g-1) and pore volume (0.74 cm3 g-1) was observed with sample HEAC-2. The adsorption efficiency of HEAC-2 was studied in vitro for paraquat as toxin using distilled water and NaCl (0.9%) solution. These study shows paraquat was adsorbed more on HEAC-2 in the presence of sodium chloride solution (4.68 mgL-1) than in distilled water (3.62 mgL-1). Furthermore, a comparision was done between HEAC-2 (4.68 mgL-1) and commercially available activated carbon (4.18mg L-1) which proved HEAC-2 is more effective than commercially available activated carbon

Luvunga scandens belong to the family of Rutaceae which usually inhabits tropical and moist environment. This plant is known as ‘Mengkurat Jakun’ among locals and used traditionally to treat fever and fatigue via decoction. The study aimed to elucidate the compounds from L. scandens stem, which were isolated using bioactivity guided isolation together with their mechanism of action on human breast cancer cell (MCF-7). The bioactivity-guided isolation of cytotoxic agent of the stem of L. scandens resulted in the isolation and characterization of a new triterpenoid from this species, 3-oxotirucalla-7,24-dien-21-oic-acid (2), along with a known triterpenoid, flindissol (1). The isolation was conducted using chromatographic techniques on silica gel and sephadex LH-20. The structures of the isolated compounds were elucidated on the basis of spectroscopic analysis including UV, IR, NMR, MS and 2D NMR. The cytotoxic activity of the plant on the proliferation of MCF-7 cell line was evaluated by MTT, WST-1 assay, scanning electron microscope (SEM), flow cytometry and RT-qPCR. The cytotoxic evaluation of the extracts showed that IC50 value of the dichloromethane (LSC-SD) and methanol (LSC-SM) extracts from the stem were 75.0 and 77.0 µg/mL respectively. Whereas IC50 value of n-hexane (LSF-H), n-hexane:DCM (LSF-HD), DCM (LSF-D) and DCM:MeOH (LSF-DM) fraction extracts were 97.5, 35, 98.9 and 94 µg/mL respectively. The IC50 value of MeOH (LSF-M) fraction could not be extrapolated since none of the concentration of this extract was able to reduce the proliferation activity to 50 %. Compound 1 and 2 showed potent cytotoxicity against MCF-7 cell line with IC50 values of 13.8 µM and 27.5 respectively after 24 h of treatment. Doxorubicin was used as a positive control drug with IC50 values of 6.21 µM. Morphological analysis of the cells surface exhibit the apoptosis results after 24 h treated with LSC-SD extract, 1 and 2, where the cells were rounded up, shrank and lost contact with neighboring cells. Apoptosis of MCF-7 cells treated with both compounds were confirmed by flow cytometry. The results show that compound 1 and 2 exert their anti-proliferative effect on MCF-7 cells through inhibiting cell cycle where the distribution of cells at Sub G1 phase was 7.7 and 9.3 % respectively. Further evident of apoptosis induction by compound 1 and 2 towards MCF-7 cell lines were assayed by RT-qPCR on the expression of PUMA, caspase-8 and caspase-9. In this assay, both compounds showed the increased of the expression of PUMA, caspase-8 and caspase-9 gene. In conclusion, both triterpenoids, 3-oxotirucalla-7,24-dien-21-oic-acid (2) and flindissol (1) have a potential to be developed as anticancer agents against breast cancer.

Type 1 diabetes is a perpetual and profound disease impacting people at all ages. It is diagnosed with the loss of insulin production because of dynamic demolition of the ?-cells in the pancreas. One bottleneck of the drug discovery on type 1 diabetes is the expensive and long testing period of the established in vivo assays. Psychotria malayana has been reported traditionally to treat diabetes. Thus, the aim of this study were to develop a type 1 diabetic adult zebrafish model through chemical induction, and to evaluate the anti-diabetic activity of Psychotria malayana Jack leaf extract on the developed type 1 diabetic adult zebrafish. Different chemical inducers, streptozotocin and alloxan, with different doses were evaluated in elevating the blood glucose level of zebrafish. Two positive controls (glibenclamide and insulin) were tested for the confirmation of the developed model. Finally, the anti-diabetic activity of a traditional medicinal plant, P. malayana aqueous extract was evaluated using the developed model. LC-MS based fingerprinting of the zebrafish and the histological examinations were applied to confirm the ability of this plant to treat type 1 diabetes. The LC-MS data set was pre-processed and then statistically calculated using a multivariate data analysis, partial least square-discriminant analysis (PLS-DA). The result of this study indicated that a single intraperitoneal injection of alloxan with the dose of 300 mg/kg was the optimal dose to elevate the fasting blood glucose level of the zebrafish. The blood glucose level (>150 mg/dL) was significantly higher (P<0.05) than that of the healthy zebrafish (70-80 mg/dL), and can be maintained for 7 days. This model was able to test the anti-diabetic activity of P. malayana leaf extract. The plant extract with the dose of 1, 2, and 3 g/kg significantly reduced the blood glucose level of the diabetic zebrafish from (123.50±10.89) to (75.67±17.82, 90.83±7.86, and 73.50±7.66, respectively) (P< 0.05). Furthermore, LC-MS based fingerprinting exhibited that all doses of the plant extract were able to shift the serum metabolite profile of the treated zebrafish toward the healthy group alongside PLS-DA component 2. While PLS-DA component 1 indicated that the plant extract with the dose of 3 g/kg was superior compared to other doses in shifting the metabolite profile toward the healthy zebrafish. Finally, the histopathological examination showed that this plant extract could not repair the enormous reduction of the Langerhans cells in the pancreas of the diabetic zebrafish. In addition, this extract did not affect the liver structure of the healthy zebrafish. Hence, the mode of action of this plant extract in lowering the blood glucose level should not be through the recovery of the pancreas, but through other mechanisms which is recommended to be examined in future research.

Anxiety disorders (AD) are increasing in all societies, especially developed ones. Caffeine (CAF) consumption in coffee, tea, and other beverages may aggravate and increase the frequency of anxiety symptoms. Vinpocetine (VP) is a synthetic derivative of the lesser periwinkle plant alkaloid vincamine and is used as a neuroprotective agent. The study was conducted to evaluate the capacity of VP to alleviate anxiety symptoms and to compare its effectiveness to that of lorazepam (LZP). The anxiety model in rat was induced via CAF injection (i.p.). Behavioural evaluation of the anxiety level in animal groups with and without treatment (oral LZP and VP) after a period of one month of treatment was done through elevated plus maze (EPM) apparatus. Additionally, neurochemical assessment of CAF, VP and LZP effects on rat brain neurotransmitters were done by using Ultra-performance liquid chromatography (UPLC). The results demonstrated the anxiogenic action of ca?eine in rats receiving a high dose of CAF (100 mg/kg), as indicated by a significant increase (p < 0.001 ): in the number of entries (29 ± 5.5) and the duration of stay (292 ± 67) in the closed arms of the EPM compared to CONT group (4 ± 1.4) and (22 ± 5), respectively. After 30 days of treatment, VP led to anxiolytic effects in EPM behaviour. A pronounced anxiolytic effect was observed in rats administered with different doses of VP (10 mg [VP1], 20 mg [VP2] and 30 mg [VP3] /kg/day; p.o.), as compared to CONT and LZP groups. The concentration of serotonin (5-HT) in cingulate gyrus of VP2 (5.45 ± 0.05), VP3 (6.06 ± 0.02) and LZP (5.1 ± 0.01) groups was significantly decreased (p < 0.001) compared to the CAF group (9.45 ± 0.18) which may indicate that the VP at moderate and high doses has a negative correlation with the concentration of 5-HT and acts as anxiolytic agent. In the hippocampus, VP has significantly increased GABA concentration in VP1 (1010.14 ± 9.4), VP3 (1043.92 ± 18.73) and LZP (1093 ± 3.35) compared to CAF group (910.56 ± 18.32, p < 0.01). The concentrations of EPI in VP2 (8.64 ± 0.79) & LZP (8.41 ± 0.03) in frontal cortex were significantly decreased (p < 0.001) compared to the CAF group (10.33 ± 0.03), which may explain the anxiolytic effects of VP and LZP on a behavioural study. On the basis of the results obtained in the present study, it is concluded that long-term administration of VP at different doses presented anxiolytic-like activity on EPM test at least via 5-HT, GABA and EPI.

The increase of antibiotic resistance has frequently been linked to the unrestrained antibiotic dispensing. A review was conducted to assess the perception and attitudes of community pharmacists towards antibiotic dispensing. The review showed community pharmacists has a proper perception on antibiotic dispensing but bad attitudes and practices. Thus a cross-sectional survey was conducted using a structured, validated and pilot-tested questionnaire. A systematic approach was used to recruit community pharmacists who completed a 27-item questionnaire in English language. Forty-five participants completed the questionnaire from sixty-five that were approached. Community pharmacists showed good perception (3.558/5) and attitudes (3.255/5) towards antibiotic dispensing. Knowledge on antimicrobial stewardship has a significant effect on pharmacists’ perceptions (p=0.034) while the number of antibiotic sold per month has significant effect on the attitudes (p=0.047). Patients visiting community pharmacies often asked for penicillin group antibiotics (80%) for cough, sore throat, cold or flu. Despite a few community pharmacists sell antibiotics without prescription, they still show a good perception and attitude towards antibiotic dispensing.

Traditionally P. macrocarpa fruits have been used as an herbal anti diabetic remedy for a long time in South-East Asia. The purpose of this study was to evaluate the cytotoxicity, acute oral toxicity and antidiabetic effects of Phaleria macrocarpa (EEPM) fruits in streptozotocin-induced Sprague-Dawley rat model. The plant fruits were extracted using 70% ethanol followed by cold maceration. Brine shrimp lethality bioassay, using sea water was prepared by dissolving 38 g sea salt in 1 liter distilled water for two days, this is allowed to hatch the shrimp to nauplii and potassium dichromate was used as a positive control. In an acute oral toxicity study, twelve male adult Sprague-Dawley rats (10 weeks) weighing 180-200 g were divided into Group-I (Control- 10% normal saline) and Group-II (extract), n=6. The fruit extract (5000 mg/kg/b.w) was given orally to each rat and observation carefully at 4 and 6 hr intervals for any physical changes. In antidiabetic study, a total of thirty-six healthy adult male rats were divided into six groups (n=6). Diabetes was induced under light ether anesthesia by a single dose (65 mg/kg/b.w) of intraperitoneally injected streptozotocin. Their glycemic status (Oral glucose Tolerance Test) was re-evaluated intermittently at 0, 30, 60, 90 and 120 min, respectively. Blood sugar level (mg/dl) and body weight of each rat in the respective groups were repeatedly measured on day 0, 7, 14, 21, 28 and 35 of the experiment. The findings of the present toxicity study suggest that the ethanol extract of EEPM fruits is non-toxic. It was found that the EEPM at 50, 100 and 200 mg/kg and glibenclamide (0.5 mg/kg) reduced the blood glucose level (hyperglycemia due to glucose load 2 g/kg p.o.) significantly after 2 hr of oral administration, when compared to the diabetic control group. The repeated oral administration of EEPM daily up to 35 days exhibited significant (p˂0.01) blood glucose activity in STZ-induced diabetic rats compared with diabetic control. At the end of 35 days of treatment, the blood glucose level of normal control, drug control and diabetic control was 132.16±5.79, 134.33±7.18 (p˂0.01) and 514.83±7.96 respectively. In the treatment groups, the dose of EEPM 200 mg/kg (392.66±3.2 to 174.33±4.3 mg/dl, p˂0.01) was shown to be more effective than EEPM 100 mg/kg (392.5±3.9 to 240.5±9.2, p?0.05) and EEPM 50 mg/kg 395.66±4.4 to 284.66±4.8 (p˂0.05). However, all selected doses showed antidiabetic activity gradually in STZ-induced diabetic rats. In histopathological examination results showed the pancreas of diabetic control were degranulated and dilated islet cells, whereas the drug control group showed granulated, nonappearance of dilation and hyperplasticity of islets. In treatment groups (EEPM at 100 and 200 mg/kg) also showed granulated pancreatic islets and prominent hyper plasticity islets. Light micrographs of rat kidney tissue in treatment groups showed various regions of the kidneys of treated animals with absence of matrix expansion and glomerular basement membrane thickening suggesting normal histoarchitecture of pancreas and renal. Biochemical aspects of the treated animal group were almost similar to the drug control group except the EEPM 50mg/kg group. In conclusion, EEPM may also serve as a good alternative in the present armamentarium of antidiabetic drugs. Keywords: P. macrocarpa, Toxicity, Antidiabetic, Histopathology, Kidney, Pancreas, Streptozotocin.

Acalypha indica is a plant that has been extensively used in the traditional medicine to treat a number of illnesses. It has been used as an expectorant, to treat skin ailments, headaches and asthma. As such, this research sought to investigate the antifungal activity and active phytochemicals of A. indica extracts and its skin sensitivity reaction. Phytochemical screening undertaken have shown that the plant contain saponins, flavonoids, steroids, terpernoids, tannins, phenolics and alkaloids. A Kirby-Bauer disc diffusion assay was undertaken for the screening of antifungal activity against Candida albicans, Microsporum gypseum and Tricophyton mentagrophytes. Both chloroform and methanol extracts have shown to inhibit the growth of C. albicans but not T. mentagrophytes and M. gypseum. Chloroform extract produced inhibition zone against C. albicans at the concentration of 50 to 200 mg/mL, whereas methanol extract produced inhibition zones at all the concentration used which was 25 to 200 mg/mL. However, the petroleum ether extract did not produce any inhibition zones against all the pathogens tested. In order to obtain the minimum inhibitory concentration (MIC) value, broth microdilution test was undertaken. The result that was achieved produced MIC value against C. albicans for both chloroform and methanol extracts at 1.56 and 0.39 mg/mL, respectively. From the result obtained, the most active extract of A. indica was continued to the skin sensitivity study. The test was conducted via Organization for Economic Co-operation and Development (OECD) guideline #406 using Buehler method and erythema readings were taken via Dermalab Combo. Guinea pigs in this study were divided into four groups which were negative control, positive control, lower dose and higher dose group. It was observed that during the challenge phase, when compared to the positive control (7.21), the erythema indices of treated groups were lower (4.11 and 4.54) which were closer towards the negative control group (3.46). Both results for week 3 of induction phase and challenge were considered significant according to the p value obtained. Active extracts of A. indica were then undertaken for fractionation and isolation via column chromatography and chromatotron. Identification of the compounds was done via the use of gas chromatography-mass spectrometry (GC-MS) and nuclear magnetic resonance (NMR) study. Identified compounds in the A. indica extracts are essential oils components such as nonacosane, and pentacosane, as well as ethyl ester (ethyl palmitate, ethyl stearate), squalene, phytol and phytosterols. The major components were the phytosterols which were the combination of three major compounds of campesterol, stigmasterol and sitosterol. The compounds discovered in this extract have been reported to possess antifungal activity against C. albicans, thus corroborating the result obtained during the antimicrobial study. Apart from that, the compounds were also found to possess other attributes such as antioxidant and also as an emollient.

Mangosteen (Garcinia mangostana Linn.) fruit pericarp has been used for centuries as a folk medicine. The study was conducted to evaluate the in vitro and in vivo antihyperglycaemic potential of G. mangostana extract (GME). The α-mangostin content in the extract was measured using HPLC and investigated for total phenolic and flavonoid contents. Antioxidant activities were measured by DPPH radical scavenging and reducing power assays whereas in vitro antidiabetic activities were evaluated by inhibition of α-glucosidase and α-amylase enzymes. Effects of GME on adipocyte cells were assessed through MTT assay, adipogenesis and glucose uptake measurements. In animal study, oral administration of GME1 (50 mg/kg), GME2 (100 mg/kg) and GME3 (200 mg/kg) to STZ-induced diabetic rats in single-dose (acute) and multiple-dose study (sub-acute) were examined. Serum biochemical parameters and histopathological alterations were evaluated and compared to standard hypoglycaemic drug, glibenclamide. The results showed that total phenolic and total flavonoid contents were 122.2±1.04 mg GAE/g and 72.8±1.75 mg QE/g of dry extract, respectively. DPPH radical scavenging activity and reducing power capacity reported with EC50 of 48.2 μg/ml and IC50 of 98.2 μg/ml, respectively. The enzymatic inhibition of α-glucosidase and α-amylase revealed higher percentage of inhibition comparable to acarbose with IC50 of 0.41 and 0.24 mg/ml, respectively. In cellular study, cytotoxicity assay reported that dose of less than 12.5 μg/ml does not affect cell viability. The differentiations of adipocytes were increased with higher GME concentration at 2.5 μg/ml, 5.0 μg/ml and 10.0 μg/ml. Glucose uptake measurements revealed a higher uptake of 2-deoxyglucose in GME2-treated cell (2243.3±232.3 cpm) as compared to GME1 (1864.0±146.3 cpm) and GME3 (1246.0±155.8 cpm). Oral administration of GME on diabetic rats indicated safe usage with absence of behavioural alterations, autonomic, neurological and toxic effects up to 2000 mg/kg. The results showed a significant reduction of glucose level in GME2 and GME3 (p<0.001) as compared to GME1. Total cholesterol, serum triglyceride, urea and creatinine were reduced in the treatment group while total protein contents were increased. Histological assessment of livers and kidneys revealed reduced lesions whereas mild regenerative activity of β-cell was observed in pancreas of diabetic rats. In conclusion, the findings demonstrated that GME could be a potential source in diabetes management owing to its antioxidant content, delayed carbohydrate digestion, induction of adipocyte differentiation, improvement in glucose uptake and antihyperglycaemic effect in diabetic rats.

The increase incidence in diabetes-and obesity-related diseases in developed and developing countries has driven serious efforts towards the discovery of adipogenic differentiation-inhibitory compounds in natural products. Mangostins and triterpenoids from Garcinia have been reported to contain a wide range of bioactivities. However, its antidiabetic and antiobesity activity has not been previously addressed. With regard to the fact, this research is designed to study the antihperglycaemic and antiobesity effects of selected phytochemicals isolated from Garcinia malaccensis and G. prainiana namely alpha-mangostin, beta-mangostin, cycloartane, friedelin and lanosterol. The compounds were tested for their antihyperglycaemic and antiobesity effects in 3T3-L1 adipocytes. In this study, we first elucidated the effects the compounds on the lipid accumulation of 3T3-L1 preadipocytes by using Oil Red O staining. The ability of the cell to uptake glucose was measured by liquid scintillation counter. Peroxisome proliferator-activated receptors gamma (PPARy), glucose transporter four (GLUT4) and leptin genes expression was determined by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). The results showed that α- and β-mangostin reduced the triglyceride accumulation in 3T3-L1 adipocytes with highest lipid inhibition activity was observed at 50 μM concentration (p<0.05) when compared to MDI (0.5 mM 3-isobutyl-1-methyl-xanthine (IBMX), 0.25 mM dexamethasone, 1 μg/mL insulin) treated cells. On the other hand, treatment with other compounds significantly induced lipid accumulation. Analyses of 2-deoxy-D-[3H] glucose uptake activities showed that all the compounds including the MDI, metformin and sodium orthovanadate (positive control) significantly (p<0.05) improved the glucose uptake when compared to the basal cell. In addition, evaluation by using the adipolysis kit (Cayman Chemicals) showed that α-mangostin and β-mangostin increases the amount of free fatty acid (FFA) release. Further evaluation by the gene expression analysis showed that α-mangostin and β-mangostin may reduce lipid accumulation by inhibition of the expression of PPARγ genes. In addition, induction of glucose uptake and free fatty acid release by α- and β-mangostin was accompanied by increasing of mRNA expression of GLUT4 and leptin. Interestingly, mature 3T3-L1 cells treated with cycloartane triterpenoid was shown to enhance PPARγ and GLUT4 gene expression and decreased leptin expression. Besides, our results for cells treated with friedelin and lanosterol shows that both compounds could stimulate adipocyte differentiation and stimulate glucose uptake in adipocytes. Since, enhance PPARγ and GLUT4 genes was found to be involved in that process, we suggest that friedelin and lanosterol also will stimulate PPARγ and GLUT4 genes expression in adipocyte differentiation and glucose uptake process, respectively. As a conclusion, these line of evidences indicated that α-mangostin, β-mangostin and cycloartane triterpenoid together with friedelin and lanosterol derived from Garcinia may become an interesting candidate for the prevention of metabolic disorders such as diabetes and obesity.

Introduction: Hypertension is one of the leading causes of death due to stroke and heart diseases, Nigella sativa “black cumin” seeds have been widely used in traditional medicine for diseases treatment including hypertension. Thymoquinone (TQ) is one of the major active constituents in its volatile oil. Objective of this study was to evaluate the antihypertensive potential of TQ and to investigate the underlying mechanisms of action. Method: In normotensive rats; mean arterial blood pressure (MAP) and heart rate (HR) was recorded using the non-invasive tail cuff technique. Dose-response relationship was obtained by using 3 TQ doses (2.5, 5 and 10 mg/kg) intraperitoneally to 3 different groups (n=5) of adult male Sprague-Dawley rats under pentobarbital anesthesia. MAP was then measured for other two animal groups pretreated either with atropine (P-at) or propranolol (P-pro) followed by 10 mg/kg TQ. Hypertension was induced in another group of animals (n=36) and control (n=6) by administration of L-Nitro-Arginine Methyl Ester (L-NAME) in their drinking water for four weeks. At the end of the induction period, rats were divided into six groups (n=8); TQ2.5+L-NAME, TQ5+L-NAME, TQ10+L-NAME, captopril+L-NAME, L-NAME only and control. MAP and HR were recorded by the tail cuff technique weekly for four weeks. Then animals were sacrificed, and blood was collected for determination of ACE activity and aldosterone concentration using ELISA. Lipid profile was assayed twice; at the end of the induction period and the end of the treatment period. Results: TQ produced a dose-dependent blood pressure lowering effect, where 2.5 5 and 10 mg/kg of TQ treatment decreased MAP by 8±1 mmHg, 12±3 and 29±3 mmHg, respectively. TQ-induced MAP reduction was significantly less in P-at than non-pretreated group. Conversely, TQ-induced MAP reduction in P-pro did not demonstrate a significant difference from the non-pretreated group. TQ reversed the established hypertension in TQ5 and TQ10 groups, and prevent further increase in MAP in TQ2.5 group. TQ antihypertensive activity was associated with a decrease in serum aldosterone concentration and an increase in ACE activity. TQ treatment lowered all the blood lipid profile parameters. Conclusion: This study confirms the dose-related hypotensive effect of TQ. The mechanism of TQ-induced hypotension involves at least in part activation of vascular muscarinic receptors, but not ?-adrenergic receptors. The antihypertensive activity of TQ takes place through renin angiotensin aldosterone system.?

Glycosmis pentaphylla (Retz.) DC. has been traditionally used to cure various illnesses and conditions such as fever, eczema and rheumatism. The present study was designed to evaluate the antimicrobial and DNA binding activities of the alkaloids isolated from the plant. Leaves and stem bark of G. pentaphylla were extracted by continuous extraction using hexane and acetone prior to acid base extraction. Screening of alkaloids available was done by Thin Layer Chromatography (TLC). Fractionation by column chromatography was employed to separate the extract into fractions of alkaloid compounds. Antimicrobial active alkaloids were screened by TLC Agar Overlay Assay against Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922 and Candida albicans ATCC 90028. The alkaloids were isolated by using column chromatography and were authenticated by comparing their TLC profile, maximum wavelength for UV absorption in methanol and melting point to that of authentic alkaloids. The identification of the alkaloids was further confirmed by their NMR spectral data. The antimicrobial activity of the alkaloids was determined quantitatively by means of their Minimum Inhibitory Concentration (MIC) and Minimum Microbiocidal Concentration (MMC) by using broth microdilution assay. Antimicrobial combination effects between the active alkaloids and selected antimicrobial agents, which were erythromycin and vancomycin was studied by cherkerboard assay and determined by their Fractional Inhibitory Concentration Index (FICI). DNA binding activity of the alkaloids was investigated by method using restriction enzymes and specially designed 1.5 kb DNA fragment. Three antimicrobial active alkaloids labeled as GP3-3, GP6-2 and GP11-2 were detected, isolated, authenticated and identified as arborinine, arborine and skimmianine respectively. Antimicrobial activity of arborinine and arborine were ranged between 250 ?g/mL and 1000 ?g/mL. Arborinine and arborine showed lowest MIC values of 250 ?g/mL and 500 ?g/mL respectively which were against C. albicans. Arborine displayed high MIC values against all other microbes while arborinine was weak against S. aureus, E. coli and P. aeruginosa. No synergistic effect was observed from the combination of the alkaloids with the selected antimicrobial agents against Gram positive, S. aureus, Gram negative E. coli and fungi, C. albicans. However, partial synergy was reported for all interactions between arborine and antimicrobial agents against S. aureus and interaction between arborinine and ketoconazole against C. albicans. Additive effect was only produced when arborine was combined with vancomycin while antagonism was observed for the interaction between arborine and ciprofloxacin when tested against E. coli. All alkaloids displayed photo activated DNA binding activity by strength in the order of arborine, arborinine and skimmianine.

Apolipoprotein E gene (APOE) has been known for more than 30 years and has been widely studied around the world for its role in the pathogenesis of diseases that are closely related to lipid and lipoprotein metabolism. Studies regarding the association between the APOE gene and type 2 diabetes mellitus (T2DM) are scarce. Therefore, this case-control study was aimed to investigate APOE allelic frequencies among the diabetic and non-diabetic subjects and associations between the alleles and selected bio-chemical markers between them. A total of 102 subjects were recruited, 51 were diabetic, and 51 were non-diabetic. Their fasting blood samples were analyzed for fasting blood glucose (FBG), total cholesterol (TC), triglyceride (TG), high density lipoprotein (HDL) and low density lipoprotein (LDL). Restriction Fragment Length Polymorphism technique was used to identify the APOE alleles. Statistical analyses were performed using the Predictive Analytics SoftWare (PASW) Statistics, Version 18.0. Ɛ2 and Ɛ4 alleles were slightly higher, and the Ɛ3 allele was slightly lower in the diabetic group compared to the non-diabetic group (12.7% vs 8.8%, 24.5% vs 22.6%, and 62.8% vs 68.6% respectively). Diabetics with Ɛ2 allele had the highest mean values of all selected bio-chemical markers, and the trend was followed by Ɛ4 and Ɛ3 alleles. Both the Ɛ2 and Ɛ4 allelic diabetic subjects had significantly higher FBG compared to the Ɛ3 allelic diabetic subjects (10.11 vs 8.18 and 9.89 vs 8.18 mmol/L respectively, p<0.05). In contrast, diabetic subjects with Ɛ2 and Ɛ4 alleles had significantly higher TC (6.57 vs 4.58 and 5.41 vs 4.07 mmol/L respectively, p<0.05) while only Ɛ4 allelic diabetic subjects had significantly higher TG (1.57 vs 0.97 mmol/L, p<0.05) compared to the non-diabetic subjects. In conclusion, the APOE gene polymorphism influences blood levels of the selected bio-chemical markers in subjects with T2DM.

Skin cancer is reported as one of the most common types of cancer with increasing numbers of occurrence. Luvunga scandens is one of the medicinal plants found in Malaysia. This plant is known to possess many bioactivities and general health effects, yet its anti-proliferative effect is generally under reported and need to be scientifically evaluated. The aim of this study is to evaluate the anti-proliferative and apoptotic effects of Luvunga scandens plant leaves against human skin cancer cell line. MTT assay was used to test the cytotoxicity effect of L. scandens on human skin carcinoma cells together with its effect against normal cell lines (HaCaT and HDF). Scratch assay was carried out to evaluate the cell growth. The morphological changes of L. scandens treated skin cancer cells was confirmed by scanning electron microscopy, and the apoptotic effect of the plant against skin cancer cells was tested using caspase 3/7 assay, followed by cell cycle analysis done using a flow cytometer on skin cancer cells. Western blot was preformed to investigate the anti-carcinogenic effect of L. scandens against human skin cancer cells. The results showed that the extract and a compound 3-oxotirucalla-7,24-dien-21-oic acid possesses cytotoxic effect against skin cancer cells, and it was no cytotoxic activity in both of HaCaT and HDF cells. The scanning electron microscopy results demonstrate that L. scandens treated cells showed an overall change in the cell shape, alteration of surface morphology, absence of microvilli and appearance of blebs. Caspase 3/7 assay results showed that L. scandens dichloromethane (DCM) extract exhibited the highest level of apoptosis against skin cancer cells. For cell cycle analysis, all the L. scandens treated skin cancer cells show high readings in the sub-G1 phase. For western blot the L. scandens extract and compound show high apoptosis effects against human skin cancer cells. This in vitro study has proven that L. scandens plant exhibit anti-proliferative effects against human skin cancer cell hence, it can be considered as a promising natural source for anticancer therapy.

Oral squamous cell carcinoma (OSCC) represents the majority of oral cancer. Chemotherapy is commonly used to treat OSCC especially as the disease advanced. However, conventional chemotherapy is associated with terrible adverse effects and the occurrence of chemoresistance which causes treatment failure. Therefore, the quest for a more effective and safer alternative has intensified. High dose ascorbic acid has been evidenced to confer anticancer effects via generation of reactive oxygen species (ROS) and through epigenetic mechanisms. Poly lactic-co-glycolic acid (PLGA) was used as the encapsulating polymer for the delivery of ascorbic acid to the cancer cells. A rapid UV-visible spectrophotometric method was validated as per ICH guideline and applied throughout this study for the quantification of ascorbic acid. Double emulsion solvent evaporation method was used to fabricate ascorbic acid loaded PLGA (AA-PLGA) nanoparticles. Optimisation of formulation was carried out by multilevel categoric full factorial design based on different surfactant concentrations and surfactant types used in the primary emulsion. The particle size of the optimised formulation was found to be 252 ± 2.98 nm, polydispersity index (PDI) of 0.151 ± 0.02, zeta potential of -20.93 ± 0.87 mV and encapsulation efficiency of 69.73 ± 1.07%. Scanning electron microscope images revealed the spherical shape of nanoparticles. The drug release behaviour exhibited a biphasic pattern namely initial burst release followed by slower release. The optimised nanoparticles formulation was further incorporated into different concentrations of Carbopol® gel. The pH of the prepared formulations was well within the pH range of the oral cavity which is between 6.2 – 7.6. Statistical analysis indicated that Carbopol® concentration significantly (p-value < 0.05) affected viscosity, spreadability and mucoadhesion of the gels. The properties of the prepared formulations were compared with a Carbopol® based commercial product (Kin Care) as a reference. Carbopol® polymer with a strength of 1% was chosen as the optimum gelling agent for AA-PLGA nanoparticles gel. The viscosity of the formulation with 1% Carbopol® was slightly higher (p-value < 0.05) than the commercial product, yet the spreadability and adhesion results were comparable (p-value > 0.05). The rheological study showed that all the gels exhibited a pseudoplastic behaviour which is desirable as it facilitates the flow of gel out of the tube and can form a viscous gel at the application site. Besides, the optimum gel formulation exhibited a zero-order kinetic release of ascorbic acid nanoparticles for 6 hours duration. Hence, it is a good candidate for topical application on the oral mucosa. The optimised AA-PLGA nanoparticles were subjected to cytotoxic assay against the OSCC SCC-25 cell line. Through in vitro cytotoxicity study, AA-PLGA nanoparticles mediated a significant (p-value < 0.05) reduction of cancer cell viability in a dose-dependent manner with an IC50 value of 2420 µg/mL. Severe cellular morphological changes were examined with an inverted microscope after 24 hours of incubation with AA-PLGA nanoparticles evidenced the cancer cell death in the SCC-25 cell line. The results of the present study support the feasibility of AA-PLGA nanoparticles gel to treat OSCC and hope the formulation can open a new avenue for OSCC therapy.

Background: Nowadays, Medication Errors (MEs) have appealed more interest of health care organizations because of their complications like higher mortality rate and increased health care cost. The MEs are underreported in all countries. This issue compromises the Medication Error Reporting (MER) systems. Objectives: This study was aimed to explore the knowledge and attitudes of nurses toward ME reporting. Furthermore, barriers and facilitators towards MER practices among nurses have also been investigated. Method: A mixed methodological approach has been adopted to answer the research questions. In the qualitative part, a total of twenty-three nurses were interviewed, using new validated semi structured interview guide. The saturation point was reached after twenty interviews. Three extra interviews were conducted to confirm saturation point. The audio recordings were transcribed verbatim and exposed to thematic content analysis. In order to generalize the initial qualitative study findings, a cross-sectional survey has been carried out with a sample of nurses attached to the same hospital (n = 310). Using new validated self-administered questionnaire. Result: Four major themes were emerged from the transcripts. Almost all the interviewed nurses were aware of the existence of MER system, MER process and the importance of MER. However, although the majority of interviewed nurses did not submit any ME report, they showed a positive attitude towards MER. The main barriers for MER were the impact of time and workload, the shame feeling, fear of being investigated and the impact of MER practices on job record, negative reaction from person in-charge, the confidentiality of the reporting form, and absence of effective feedback. The obtainable response rate was equal to 90.3%. The mean for their total knowledge score was 5.6 (SD= 1.4). Almost all participants considered MER is important (99.3%) but 30 % of those who committed ME last year did not report their errors. Almost all the respondents agreed that MER is part of their professional duties (92.1%). The main reasons for underreporting were fear of liability or lawsuit issues (63.6%), fear of being labelled as an incompetent nurse (52.1%), and lack of effective feedback from the nursing manager (33.6%). The main facilitators toward MER were compulsory MER (90.7%), conducting training program specifically for MER (84.3%), analysis/feedback criteria (80%), encouragement from nurse leaders (75.7%), and using anonymous reporting form (73.2%). Conclusion: This study revealed that although the nurses had reasonably acceptable knowledge and positive attitudes toward MER, there was low involvement of nurses toward MER. Their decisions to report MEs were highly affected by barriers. In order to improve the participation of nurses toward MER, the related authorities should work on strategies which can help to improve the involvement of nurses such as creating a MER environment free from blaming, reporting anonymously, and conducting a training program.

Attitude of healthcare professionals gives big influence towards self-esteem of people with disabilities (PWD). The greatest challenges faced by PWD were not being treated with respect, not equipped with suitable equipment, and denied health services. This study aimed to identify general attitude of healthcare professionals towards PWD and to explore this attitudes from the perspective of PWD. A mixed method approach was conducted involving 350 healthcare professionals from Hospital Tengku Ampuan Afzan (HTAA) and 10 physically disabled people who used or still receiving treatment in HTAA in a cross-sectional survey and face-to-face in-depth interviews, respectively. A Scale of Attitudes toward Disabled Persons (SADP) was distributed to respondents and data were analyzed using IBM SPSS Statistics version 20. On the other hand, data from face-to-face in-depth interviews were analyzed using thematic analysis.The quantitative findings revealed physicians have the most favorable attitude followed by optometrists. Meanwhile the least favorable attitude were shown by medical assistant followed by nurses. From the perception of PWD, majority of the participants were satisfied with positive attitude shown by healthcare professionals though negative attitude was claimed to exist in some situations. With respect to gender, from quantitative findings, males were significantly having more favorable attitudes compared to female (p=0.002). Meanwhile, from in-depth interviews, the participants mentioned that there was no difference in attitude shown with regard to gender. Besides, in terms of age and working experience, quantitative findings revealed weak negative correlation between age and working experience in relation with attitude towards PWD. However, from the qualitative findings, the participants mentioned about older healthcare professionals are advance in terms of their knowledge and experience. Six themes were emerged from saturated data namely: Being as PWD, sensitivity and care, challenges encountered, support towards PWD, health professionals’ attitude towards PWD, hospital’s facilities and services provided for PWD, and implications towards PWD. Majority of the participants expressed to feel demotivated and having low self-esteem after having their impairment. Support from close family members and friends are really significant to boost their self-esteem and motivation. Problems encountered in healthcare setting should be overcome in order to ensure PWD are able to adherence to appointment and rehabilitation sessions.

E-learning is considered one of the major trends in the digital age. Online learning helps pharmacy students and pharmacy practitioners to continue their professional development and to improve their skills via the internet. E-learning has positive effects on medical and pharmacy education in developed countries. Educators nowadays can build online educational materials based on their students learning styles and manage the learning process through learning management systems. However, scarce research efforts have been observed about the effectiveness of e-learning and web 2.0 tools in pharmacy education regarding transferring of pharmaceutical knowledge in Arabic Language Speaking Countries (ALSCs). This study consolidated into two phases. In the first phase a cross-sectional study design has been adopted to evaluate the attitudes and perceptions of Arab pharmacists and pharmacy students. While one-group pre-test post-test study design has been used in the second phase to assess the effectiveness of using e-learning and web 2.0 tools and explore the views of study participants regarding the effectiveness of using these technologies in transferring of pharmaceutical knowledge. The study results showed that the majority of respondents have highly positive attitudes towards using e-learning and emerging web 2.0 tools in transferring pharmaceutical knowledge. Similarly, the majority of them have high confidence level in learning and shared the same motivation to learn using such novel tools. Using Schoology® as LMS and Facebook® as the most popular social network site was effective. Upon completion of the online course, a significant improvement in the participants’ level of knowledge about the studied topic has been observed. Highly positive views of respondents regarding the effectiveness of e-learning and web 2.0 tools have been perceived. Major logistical and technical barriers imped the successful adoption of e-learning in ALSCs. However, this study showed a promising results regarding using novel online educational technologies in transferring of pharmaceutical knowledge. Further researches are encouraged to study the convenient, compatibility and the effectiveness of e-learning and web 2.0 tools in pharmacy education in ALSCs. A steady expansion in the digital gap between ALSCs and developed nations empowers launching well-planned initiatives to facilitate the proper use and implementation of novel technologies in pharmacy education

Coronary heart disease (CHD) and stroke are the major causes of mortality in Malaysia. According to the WHO, the mortality rate of CHD within Malaysia is increasing and was responsible for 23.10% of total deaths in 2014. The high rate of mortality due to heart attack and stroke is attributed to lack of awareness towards their signs and symptoms. Therefore, this study aimed to assess the awareness and response towards signs, symptoms and risk factors of HA and stroke among the general population at Kuantan, Pahang, Malaysia. A cross-sectional study was conducted via structured questionnaire survey among 393 participants in Kuantan city who were between 18-64 years of age. Majority of the respondents recognised chest pain as heart attack symptoms (HAS) followed by 68.1% sudden shortness of breath. Only 35.6% of respondents reported suitable action towards HAS. About 81.9% knew at least one HAS while 10% of them did not know any HAS. Approximately 11.5% of participants identified all five HAS and 5.6% of participants recognised all the five HAS and appropriate action (calling an ambulance). Multivariable logistic regression showed that single participants were more aware of all 5 HAS than other marital statuses (p=0.023, OR=0.023, 95% Cl=0.001-0.594). Malay participants were more likely to recognise all the 5 HAS than other race (p=0.004, OR=0.376, 95% Cl=0.193-0.773). Respondents with family history of HA, those who had received information about HA and those who were aware that HA needs urgent treatment showed more awareness on all 5 HAS than others without those conditions (p=0.055, OR=2.206, 95% Cl=0.983-4.949), (p=0.002, OR= 7.540, 95% Cl=2.0337-27.914), (p=0.01, OR= 0.17, 95%=0.044-0.710), respectively. Furthermore, the majority of individuals recognised smoking as risk factors of heart attack (RFOHA), while 91.2% of them recognised at least one RFOHA and 5.6% of individuals identified all modifiable risk factors. With regards to stroke, the majority of participants (78.6%) recognised sudden numbness or weakness of the face, arm or leg as symptoms of a stroke (SOS), followed by 74.6% of them who identified sudden trouble walking dizziness, loss of balance as SOS. Only 29.8% of respondents were aware of appropriate action towards signs and symptoms of a stroke by calling an ambulance. Approximately 88.8% of individuals classified at least one SOS while 11% of them did not recognize any SOS. Moreover, 27.5% of the participants were aware of all five SOS, while 9.4% of them identified all five SOS and appropriate action. Additionally, majority of the respondents (69.2%) were aware of hypertension as stroke risk factors (SRF) and 89.2% of those identified at least one modifiable stroke risk factors (MSRF). About 9.2% of them know all MSRF. Multivariable logistic regression showed that participants aged 18-45 years were more likely to be aware of all SOS (p=0.01, OR=0.282, 95% Cl=0.083-0.963). Hypertensive participants were more likely to be aware of all SOS (p=0.01, OR=0.129, 95% Cl=0.025-0.673). The awareness and action towards HAS, SOS and its risk factors among the general public in Kuantan were poor. Participants with high education and income represented better awareness toward HAS, SOS and its risk factors than those with low level of education and income. The awareness of HAS, SOS and its risk factors is important to reduce pre-hospital delay and mortality. Educational intervention is, thus, recommended to raise the awareness of HAS and SOS as well as to reduce the mortality due to HA and stroke.

Entada spiralis Ridl. known as Sintok or Akar Beluru is the woody climber plant belong to Leguminoceae family. E. spiralis is an excellent foaming agent due to the presence of saponin. Thus, E. spiralis stem bark is established used as natural body soap, shampoo and washing agent. The possession of antioxidant activity has been revealed by E. spiralis ability to treat syphilis, insect bites and superficial skin diseases. This research aims to screen the major compounds by phytochemical screening test, verify the antioxidant compounds from crude extracts and fractions via in-vitro antioxidant activity assays, isolate the antioxidant compounds through chromatographic methods and characterize the structure of antioxidant compounds by using various spectroscopic techniques. The crude extract of E. spiralis leaves was prepared by macerating the leaves with petroleum ether, chloroform, and methanol solvent sequentially. Determination and evaluation of antioxidant activity was done through in-vitro study by using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, and 2,2-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical cation. Folin-Ciocalteu’s reagent method was used to measure total phenolic content and flavonoid content was evaluated by calorimetric method. The crude extract of E. spiralis leaves was fractionate by vacuum liquid chromatography (VLC) and liquid-liquid extraction. Isolation of antioxidant compounds was further with column chromatography technique and characterized by spectroscopic analysis. Methanol, chloroform, and petroleum ether crude extracts were analyzed with 5.53 %, 2.05 % and 2.55 % yield. Saponin and flavonoid were the major components. Methanol extract exhibited the highest antioxidant activity with IC50 (DPPH); 40.23 ± 2.66 ug/mL and IC50 (ABTS); 5.09 ± 0.53 ug/mL. The highest possession of phenolic content and flavonoid content; 124.67 ± 6.63 mg GAE/g and 51.67 ± 2.17 mg QE/g were also displayed by the methanol extract. Isoscutellarein, kaempferol-3-C-rutinoside, kaempferol-3-O-glucoside, and kaempferol glycoside were isolated from liquid–liquid extraction fraction while stearic acid and ethyl oleate were isolated from the fraction of vacuum liquid chromatography. IC50 (DPPH) of isoscutellarein; 112.18 ± 2.21 ug/mL, kaempferol-3-C-rutinoside; 136.04 ± 0.52 ug/mL, kaempferol-3-O-glucoside; 301.01 ± 3.02 ug/mL, and stearic acid; 187.49 ± 1.37 ug/mL. Ethyl oleate and kaempferol glycoside were showed negative results towards DPPH. Whereas, IC50 (ABTS) of isoscutellarein; 17.50 ± 0.26 ug/mL, kaempferol-3-C-rutinoside; 34.10 ± 1.13 ug/mL, kaempferol-3-O-glucoside; 14.65 ± 0.11 ug/mL, stearic acid; 5.69 ± 0.06 ug/mL, ethyl oleate; 474.37 ± 2.91 ug/mL, and kaempferol glycoside; 124.82 ± 6.60 ug/mL. In conclusion, the active crude extract and fraction was verified by several in-vitro antioxidant assays. Six antioxidant compounds were successfully isolated from this research study.

The work presented in this thesis describes selected biological activities of the salivary gland secretion of the medicinal Malaysian leech Hirudinaria manillensis (Lesson, 1842). A new device for leech feeding was developed using common laboratory tools consisting of a glass funnel wrapped with a parafilm membrane and filled with the phagostimulatory solution. Many phagostimulatory solutions were examined and only those containing sodium chloride and arginine were accepted by leeches. It was found that 71% of the tested leeches continued sucking for 15-45 min and 86% of them experienced 1-5 folds increments in body weight. For collection of a high quality and quantity LSE, we starved leeches for three weeks. A new method for LSE collection was described. The fed leeches were forced to regurgitate whatever they sucked by immersing them in ice containers. By using this method, leeches stayed alive and regain their activity after 15-30 min. The maximum concentration (A280 = 0.342) was provided within the first five minutes of sucking action. Gel electrophoresis (SDSPAGE and Tricine-SDS-PAGE) methods revealed the existence of various peptides and proteins with molecular weights ranging from 2.3-250 kDa. Amidolytic activity assay showed that LSE inhibited thrombin-induced release of p-nitroanilide from the synthetic substrate S-2238 with IC50 of 49.391±2.219 ?g/ml. LSE was shown to prolong thrombin time in vitro. The maximum antithrombin activity was obtained during the dry season (IC100 =16.081±0.079 ?g/ml). A longer starvation period yielded a lower antithrombin activity. An optimization of lyophilization conditions revealed that pre-freezing at -20