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Leaves of Coleus blumei (two species), Coleus amboinicus (two species). Coleus aromaticus and Pagostemon cahlin from Lamiaceae family were collected from different localities, freeze dried and extracted with aqueous methanol. The biological activity in vitro, especially in relation to total phenolic & flavonoid contents, antioxidant and antimicrobial activities were studied. Total phenolic content was determined according to the Folin-Ciocalteu method whilist in antioxidant activity was assessed using 2,2-diphenyl-l-picrylhydrazyl (DPPH) method. The antimicrobial activity of the extract was determined by making use of macro dilution and disc diffusion methods on two gram positive bacteria, two gram negative bacteria and on fungus. Furthermore, the toxicity was also assessed for the extracts by performing acute toxicity test. The phenolic content among the six Lamiaceae leaves extract showed significant difference (p<0.05) in result ranging from 55.21 - 95.17 mg GAL/g of dried samples. Pogoslemon cablin (PCM) had the highest content of phenolic followed by C. blumei (CBPM). There were significant differences (p<0.05) of ICso value of six Lamiaceae leaves extract ranging from 10.5 - 34.1 U-g/ml. Among the species studies. Coleus amboinicus - Malaysia (CALM) and Pogoslemon cablin (PCM) showed high antioxidant activity compared to the other leaf extracts. All leaf extracts showed activity at least against one strain of bacteria and result showed significant difference (p<0.05) between activities on the five microorganism. On the contrary, all of the leaf extracts were not effective against C. albicans. The minimum inhibitory concentration (MIC) of all leaf extracts ranged from 1.0-2.0 mg/ml in inhibiting the growth of S. aureus., E. colt, P. aeruginosa and B.subtilis. The acute toxicity test using C. Blumei leaf extract showed that there wras no mortality of animals experimented recorded even at the highest dose level of 5000 mg/kg body weight and observed for the 24 hours after administration. This shows that Coleus blumei plant extract have no toxic effect in mice. This study shows that the extracts can be used as antioxidant and antimicrobial agent without having the toxic effect. ii

The prevalence and suffering of diabetes and obesity has been increasing among the various communities of the world including Malaysia. The cost of treatment is also rising. Therefore, it is important to explore non pharmacological regime that are noninvasive and with less health risks and cost burden to the patients, healthcare professionals and nations. Medicinal plants have been used for the treatment and prevention of diabetes since ancient time. Therefore, nine common traditional antidiabetic herbs namely Andrographis paniculata (Hempedu bumi), Lagerstroemia speciosa (Banaba/bungur), Orthosiphon stamineus (Cat whisker), Peronema canescens (Sungkai), Momordica charantia (Bitter gourd/bitter melon), Tinospora crispa (Patawali), Pithecellobium jiringa (Jering) and spices namely Syzygium polyanthum (Bay leaf) and Cinnamomum zeylanicum (Cinnamon), were screened for their antidiabetic properties in in vitro model. Water extracts of these herbs and spices were prepared and evaluated for their effects on cell proliferation, adipogenesis, adipolysis, glucose uptake and glucose oxidase assay in 3T3-L1 preadipocytes. The study was then continued with quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) for selected herbs and spices extracts for the genes; adipogenesis-regulator (Ppar? mRNA), insulin-sensitive glucose transporter 4 (Glut4 mRNA) and gene associated with obesity and insulin resistance (Adiponectin mRNA). The results of aforementioned extracts promoted cell proliferation at a concentration of 0.25mg/ml which showed a maximum viability after 48 hours of treatment. Insulin and A. paniculata extract significantly (p<0.05) induced adipocyte differentiation, inhibited lipolysis and stimulated glucose uptake/oxidase in adipocytes. This activity was accompanied by significant (p<0.05) up-regulation of Ppar?, Glut4 and Adiponectin transcriptional levels. This finding reveals that A. paniculata extract have similar effect to that of insulin activity. Whilst the study on C. zeylanicum and O. stamineus extracts have similar activity for adipogenesis significantly (P<0.01), stimulated glucose uptake and reduced adipolysis activity. Thus, it is suggested that these extracts might have insulin-mimicking effects which could be potentially used as preventive agents for diabetes. In contrast to insulin, water extracts of L. speciosa did not induced adipocyte differentiation, significantly (P<0.01) decreased Ppar? mRNA, exhibited adipolysis activity and stimulated glucose uptake/oxidase due to significant (P<0.05) up-regulation of Glut4 transcriptional levels in adipocytes. This combination suggested that L. speciosa extract may be useful for the treatment of hyperglycemia and obesity in type 2 diabetes. It is well known fact that an appropriate balance between the adipogenesis, adipolysis and glucose uptake/oxidase in diabetes is of primary importance. The present study provides some important clues both on the biochemical and transcriptional aspects of the effect induced by the herbs and spices screened. The present study suggests that these herbs and spices possess antidiabetic properties as well as can be used for the associated metabolic dysfunctions.

The purpose of this study is to explore the potential of physicochemical and antioxidant characteristics of Baccaurea angulata fruit juice and to determine an acceptable dose to be consumed. Size of B. angulata fruits was measured and separated into whole fruit, berry and skin portions. Samples were blended, filtered and freeze-dried. Lyophilized juice was analyzed for nutrient and chemical compositions, with antioxidant capacity quantifications. Results showed freeze-dried skin (FDS) had the highest moisture (21.04 %) and ash (10.04%) content, protein (2.12%), total fat (2.09%), and water activity (0.467 aw), compared to freeze-dried whole fruit (FDWF) and freeze-dried berry (FDB). FDWF had the highest carbohydrate (74.12%) and total dietary fibre (6.3 %) content while FDB had highest content of crude fibre (0.36%) and gross energy (304.09kcal/100 g) as compared to other samples. Meanwhile, FDS showed highest value in diphenyl-picryl-hydrazyl (DPPH) assay (102.66mg AA/100 g) and trolox equivalent antioxidant capacity (TEAC) assay (847.46mg TE/100 g) values. Toxicity study on normal rats was carried out using FDWF sample. Animals in both acute and sub-chronic studies demonstrated no significant changes in general behaviour, growth, and relative organ weights. A significant increment of red blood cells (8.93x106/μL) was observed in low dose female group with decrease of white blood cells (5.67x103/μL) and lymphocyte (4.33x103/μL). In liver function test, total protein and alkaline phosphatase were markedly increased in male treated group (300 mg/kg), 36.50g/Land 47.90U/L, respectively. The low dose female group also showed decrement in same parameters, 63.70g/Land 73.50U/L. However, gross and microscopic appearance of the organs showed no significant pathological changes. There was no evidence of any tissue injury. In acute study, lethal dose (LD50) for FDWF was determined to be >5,000 mg/kg. Therefore, the no-observable adverse effect level for B. angulata was 1,200 mg/kg administered orally for 13 weeks. In conclusion, B. angulata whole fruit juice has the potential to be utilized for preparation of a health drink as it is safe and offers opportunity to be developed for nutraceutical and functional fruit product.

Cardiovascular diseases (CVD) have become a leading cause of death and disability and staggering increase in its incidence has been documented worldwide. Previous conducted research has provided the evidence on effectiveness of antioxidative properties of polyphenols contained in fruits and vegetables at combating various chronic diseases including CVD. Baccaurea angulata, locally known as 'belimbing dayak‘ or 'belimbing hutan‘ is one edible fruit species restrictedly distributed in Sabah and Sarawak. This present study was aimed to provide scientific data on nutritional information and health benefits of B. angulata juice extract (BAE). Pearson‘s correlation was used to obtained corelation whereas analysis of variance (ANOVA) was used to test the mean different among data. Freeze dried BAE consisted of high carbohydrate followed by moisture, total dietary fiber, ash, and protein. The extract holds high energy density with low water activity (Aw) value for storage stability. Phytochemical screening revealed that BAE contained of 7.90 0.05 mg gallic acid equivalent (GAE)/g of total phenolic acids and 12.74 0.24 mg quercetin equivalent (QE)/g of total flavonoids content and both were strongly correlated to ferric reducing antioxidant power (FRAP) value (p<0.01) but not to superoxide anion/xanthine oxidase dismutase (XOD) scavenging capacity. Male Sprague Dawley rats were induced with high cholesterol diet (normal rat chow enriched with 1 % cholesterol and 0.2 % cholic acid) and supplemented with BAE given orally for 10 weeks. No significant changes in body weight, heart and kidney weight were notified. Liver weight was significantly higher in groups consuming high cholesterol diet (p<0.05) than in normal control with slight decrease in groups supplemented with BAE than hypercholesterolemic control (CP). Plasma and liver total cholesterol (TC), triglycerides (TG), low density lipoprotein cholesterol (LDL-c), and atherogenic index (AI) were markedly increased in CP group with significant values in TC, LDL-c, and AI (p<0.05) and reduced in high density lipoprotein cholesterol (HDL-c). Reduction of plasma and liver TC, TG, LDL-c and AI were observed in dose-dependent manner upon consumption of 100 (TL), 300 (TM) and 500 mg (TH) of BAE/kg body weight/day with significant reduction in plasma TG (p<0.05). Notable increase in plasma HDL-c was found in all groups consuming BAE compared to CP group but liver HDL-c was less affected. The highly reduced plasma and liver glutathione peroxidase (GPx) and superoxide dismutase (SOD), in CP group were raised in all groups consuming BAE especially by TM group (300 mg BAE/kg bodyweight/day). Remarkably, with reference to normal control, plasma SOD value was successfully restored in TM and TH groups. In determining the effects of post BAE administration towards liver and kidney, no significant changes (p>0.05) in aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), total protein, albumin, urea, and creatinine levels were evaluated. Notable increased in AST, ALT, and ALP were observed in CP group due to hypercholesterolemic condition and with reference to normal control, these parameters were least affected in TM group. Furthermore, BAE showed no significant effects towards haematological parameters.

Malaysia is exploring opportunities in developing its poultry vaccination programme to produce better poultry vaccine to fight against the two most important diseases of poultry in Malaysia which is Newcastle disease (ND) and infectious bursal disease (IBD) which have been causing constant economic losses to the national livestock industry. Since the commercially available vaccines are consisting of less virulent virus strain that differs from the virulent outbreak strain, the safety and efficacy of the vaccines are becoming great concerns. Development of vaccines consisting of recombinant protein that contains epitopes which able to induce neutralizing antibodies are dominating in the strive for an ideal vaccine, being safe and cheap. Previous studies have shown that the viral surface proteins from Newcastle disease virus (NDV) and infectious bursal disease virus (IBDV) contains epitopes which able to induce neutralizing antibodies against ND and IBD respectively. In this study, viral protein 2 (VP2) from IBDV and hemagglutinin-neuraminidase (HN) from NDV were isolated from IBDV and NDV of local isolates via RT-PCR and cloned into pCR2.1TOPO vector. Subsequently, two constructs of recombinant plasmid containing fused gene was constructed in which full length HN gene from NDV was fused to VP2 of the infectious bursal disease virus (IBDV) while the other construct used partial HN gene. Production of the fused protein was attempted in Pichia pastoris using pPICZαC but was not successful. However, an intact fused protein of VP2-PHN constructed form VP2 and partial HN in pRSETB vector was successfully produced by the Escherichia coli. A protein band with expected molecular weight of 75 kDA was observed in SDS-PAGE and Western blot analysis upon detection with anti-Histidine monoclonal antibody. The VP2-PHN protein produced could be a potential candidate as recombinant subunit vaccine against both IBD and ND upon single immunization.

Metal Injection Moulding (MIM) is a revolutionary new class of medical implant fabrication in orthopaedics. This technology employs natural resources and holds great promises in producing large quantities of metallic parts for prosthetic implant at minimum outlay without compromising its quality. The present study was conducted to compare potential effects of MIM plate for internal fixation in fracture healing in New Zealand White (NZW) rabbits with conventional machining plate (Synthes®) as control. Following research approval by IIUM Ethics Committee (IIUM/305/20/4/10), forty rabbits were used and randomly divided into two groups. All rabbits underwent single transverse mid-shaft tibial fracture surgery and fixation under anaesthesia. The fractures were fixated with either the MIM plate or conventional plate (Synthes®). The monitoring of fracture healing was carried out at week 3, 6, 9, 12 and 26 according to ISO10993-2:2006 & ISO10993-6:2009 standards. The status of bony union was examined by means of plain radiographic assessment, macroscopic observation and, histological evaluation. There were callus formations in both groups. Bony union was evident at week 6 post-operatively, whilst bone remodelling was completed at week 26 as indicated by plain radiograph assessment and macroscopic evaluation. Histological assessment showed that both groups possessed mild to moderate callus bridging at week 3 and week 6, respectively. Complete remodelling of cortical bone was evidenced at week 26. The plate was neither broken nor bent during the study. These findings indicate that the MIM plate has equal potential to hold the fracture as the conventional plate. Therefore, the locally manufactured MIM plate has the potential to be used as an alternative internal fixator for fracture management in orthopaedic considering the value and conservation of natural resources.

The search for natural anticancer remedies is one of the most prominent research in the cancer treatments. For that purpose, various plants from all over the world have been studied. The researchers are now more focusing to understand the mechanisms involved in the cancer treatment using the natural sources. Melastoma malabathricum Linn is known as a shrub that wildly grows in the tropical and subtropical regions. A number of studies have been conducted on this plant, but the reports on its anticancer properties are still limited. The main objective of this study is to study the effects of M. malabathricum extracts from different parts at various concentrations on three types of cancer cell lines in vitro (A375, HeLa and MCF-7) and its relation to the apoptosis mechanism and the expression of the target protein, MMP-13 in the treated cells. Liquid-liquid extraction protocol using methanol, petroleum ether and chloroform as the solvent systems were carried out on leaves, stems and flowers of M. malabathricum. Dimethyl sulfoxide, DMSO (1 %) was used in the extract dilution and serial dilutions were conducted to obtain eight different extract concentration, ranging from 0.078125 µg/mL to 10 µg/mL. The evaluation of cancer cells growth inhibition for 24, 48 and 72 hours of treatment was determined using MTT assay. The treated cancer cells were then tested for morphological apoptosis detection through TUNEL assay and detection of MMP-13 expression using Western Blot analysis. The result showed that petroleum ether extracts of stems (PeMMS) and leaves (PeMML) have the best growth inhibitory effects on A375 (EC50 = 0.185 µg/mL at 48 hours) and HeLa cell lines (EC50 = 0.368 µg/mL at 48 hours). The chosen extracts were also confirmed do not cause toxicity effect on normal human fibroblast cell lines (CCD-1090Sk). Further analysis revealed that PeMMS and PeMML caused a high percentage of apoptotic cells, around 26 % in A375 and HeLa cells respectively. The apoptosis results are comparable or in approximate with the percentage of apoptosis induced by commercialized anticancer drug, paclitaxel (26.4 %). Western blot analysis showed the reduction of MMP-13 expression in the A375 and HeLa cells treated with PeMMS and PeMML, respectively. Based from the outcomes, this study suggests that the reduced expression of MMP-13 correlates with the increasing level of apoptosis in the cells treated with PeMMS and PeMML. In conclusion, the petroleum ether extract of stems and leaves of M. malabathricum showed a promising anticancer properties toward skin melanoma and cervical cancer cell lines. Further investigation needs to be conducted in order to assess the probable bioactive compounds in the petroleum ether extract of M. malabathricum that may contribute to the significance cytotoxic effects to the respective cell lines.

Collagenases, which are proteins from the matrix metalloproteinases family, have been implicated to play a crucial role in tumor invasion. Previous studies have also suggested that collagenases may have a significant influence in the expression of CTCF and YB-1 proteins, which have also been implicated in numerous cancer studies. However, the depth of its involvement in the fundamental molecular mechanisms of cancer remains to be poorly known. This study aims to determine the degree of MMP-1, MMP-8, MMP-13, YB-1 and CTCF protein expressions in breast, cervix and skin human cancer cell lines and to ascertain its probable involvement in cancer development. An ELISA assay was conducted in order to estimate the concentration of the proteins that could be found in the cells. The quantitative expressions of MMP-13 proteins in A375, MCF-7 and HeLa cells were evaluated through ELISA test. The results showed a marked expression of MMP-13 proteins in MCF-7 (12900 pg/mL), followed by HeLa (8109 pg/mL) and A375 (7515 pg/mL). This correlated to the Western blot assay where MCF-7 showed the strongest protein expression followed by HeLa and A375. The qualitative expressions of the proteins in cervix cancer (HeLa), breast cancer (MCF-7), skin melanoma (A375) and normal fibroblast (CCD1090Sk) cells were demonstrated through Western Blot analysis. The results showed that the strongest expression for MMP-1 was detected in HeLa cells at ~52 kDa, while MCF-7 and A375 cells produced very weak or almost non-detectable protein expressions. In contrast, MMP-8, MMP-13, CTCF and YB-1 proteins were highly expressed at varying degree of expressions in all cell lines. For MMP-8 analysis, multiple bands were detected at ~70 to 45 kDa, while MMP-13 only produced a single band at ~48 kDa. The results also showed the detection of CTCF protein bands ranging from ~180 to ~80 kDa and YB-1 protein expressions at ~55 kDa. In Co-Immunoprecipitation assay, results showed that most of the protein-protein interactions caused the expressions of most of the proteins to be highly up-regulated though some of the interactions also resulted in a decreased in protein expression. It was taken note that although the molecular mass of the proteins correlated to its respective Western blot assay results, their protein expressions were either highly expressed or were of less prominence. It was elucidated that the increase and decrease of the protein expression may have been caused by the interactions and binding of the proteins to its protein partners which then affect cellular regulation. Overall, although variations were found in the expression of the collagenase in various human cancer cell lines, the results depicted a prominent presence and the probable involvement of MMP-1, MMP-13, YB-1 and CTCF proteins in the development and metastasis of cancer. Thus, it is believed that obtaining a detailed understanding of collagenases and its interacting partners may be vital for future endeavours in respect to the possible design of therapeutic MMP strategies for prospective targeted human therapeutic treatments.

The research application for drugs and food supplements derived from plants extracts have increased in recent years. Plant extracts and their constituents are recognized to be safe, either because of their traditional use without any documented detrimental impact or because of dedicated toxicological studies. On the other hand, human resistant pathogenic microorganisms increase worldwide mortality and morbidity. Due to emerging resistant and low susceptible strains to antibiotics, search for new alternative antimicrobials is raised significantly. Medicinal plants had been investigated and had been considered as potential sources of antimicrobial agents. Although hundreds of plant species have been tested for antimicrobial properties, the vast majority of medicinal plants have not been adequately evaluated. Thus, a systematic investigation was undertaken to screen for antibacterial activity from Baccaurea angulata (BA). The purpose of this study was to explore bacteriostatic and bactericidal effect of methanol, ethanol and aqueous extracts of three parts (whole fruit, fruit skin and berry) of BA fruit in vitro against tested pathogenic microorganisms. Total 9 extracts from three parts of BA fruit (whole fruit, fruit skin and berry) were tested against Gram positive (Staphylococcus aureus, Streptococcus pneumonia, Staphylococcus epidermidis, and Clostridium botulinum) Gram negative bacteria (Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae and Salmonella paratyphi A), and fungus (Candida albicans), using disk diffusion, agar well diffusion and microdilution methods. Results show that BA fruit extracts have potential antimicrobial properties against all tested microorganisms, it is various between three parts (whole fruit, fruit skin, and berry), three solvents (methanol, ethanol and aqueous), different method (agar well diffusion, disk diffusion and microdilution method) and different aforementioned pathogens. The highest inhibitory activity was observed in ethanol extract of fruit skin using agar well diffusion against Streptococcus pnumoniae (37.0±1.0 mm) at the concentration of 1,000.0 μg/mL. Among tested Gram negative bacteria K. pnumoniae was the most susceptible bacterium. The extracts showed minimum inhibitory and minimum bactericidal activity at the concentration of 7.8 and 15.6 μg.mL respectively using microdilution method.

This study was conducted to evaluate the severity of medium filth contamination in ready to eat food (RTE) as to confirm the definition of halal food that supposedly not contaminated with najs mutawassitah. A total of 52 human stools samples were collected from voluntary healthy subjects according to method as explained by Chessbrough (1987) and the screening of bacteria in the human stools samples were done according to traditional microbiological analysis methods. Determinations of bacterial growth curves were performed using NanoDrop 1000 UV-VIS Spectrophotometer at 630nm where the initial and end of lag times for each of bacteria was determined. The growth evaluation of faecal borne bacteria in RTE food was performed using prepared fried rice samples. The prevalent study of food-borne/faecalborne bacteria was performed in 120 RTE fried rice collected from four different types of food premises in the town of Kuantan, Pahang. The results showed that healthy human stools which fall under najs mutawassitah contained high amounts of presumptive pathogenic bacteria specifically E. coli, S. aureus, B. cereus, Aromonas spp. and Salmonella spp. at different mean values. Total plate count (TPC), coliform and F. coliform were used as indicators in detecting the presence of pathogenic bacteria in human stools as well as for contamination of najs mutawassitah. Average lag phase time for faecal borne bacteria was around 60 minutes. Thus consumption of food within one hour should not give any significant health effects. Consuming food which contains faecal borne bacteria within one to two hours would give either low risk health effect or none at all. Consuming food after two hours has medium risk. Consuming food after three or four hours has the highest health risk. If the contamination of human stools in the food is in small quantity (1-2 drops), it may have no health risk at all. The small amount of bacteria in food may need more time to adapt with the new environment. If the human stools are in higher volume (more than 2 drops or about 1 ml) then it will start to contaminate the food and could then lead to health risks. If RTE food were contaminated with small amount (about 0.1 ml) of human stools and were left over at ambient temperature (about 37oC) for a certain period of time (about 4 hours), it would start to have bacteria contamination and may cause health risks. If the level of health risk was translated according to Shariah law, RTE foods which were contaminated with higher amount (more than 2 drops) of human stools or contaminated with small amount (1-2 drops) of human stools and were left exposed at ambient temperature for more than 4 hours can be considered as shubhah/makhrooh to be eaten. The study also indicated that RTE fried rice sold at markets have medium to high health risks. Fresh or just cooked fried rice which are sold at night markets have less health risks compared to those that are sold at other type of food premises.

Infectious Bursal Disease Virus (IBDV) outbreak had been reported to infect broiler chicken in Malaysia and causes high mortality. It had been reported that the virulence and pathogenicity of the virus is related to its coat protein, more specific the VP2 hypervariable regions. The study aimed to determine the mutation occurred in the amino acid sequence which responsible for the outbreak of the disease and increased in the virus virulence. To study this, the nucleotides and protein of the local IBDV isolate was sequenced and analyzed by comparing it with other IBDV that had been reported previously. To obtain the virus sequence, local IBDV isolate strain 3529/92 was propagated in Specific Pathogen Free eggs and RNA was extracted and amplified by RT-PCR before being inserted into pCR2.1 TOPO TA vector for cloning and sequencing. The full length of the gene segment of the coat protein was constructed by concomitantly joining the fragments using the IBDV native restrictions sites in E. coli expression vector, pTrchis2a. The sequence analysis revealed that the local IBDV 3529/92 consists of 3039 nucleotides which encodes for 1012 amino acids. BLAST analysis of the nucleotide sequence showed that the local strain shared the greatest similarities with Dutch D6948 very virulent (vv) IBDV subtypes. Analysis of the VP2 variable regions of 3529/92 showed most of amino acid substitutions occurred at VP2 variable region are very similar to the changes in VP2 variable regions of the vvIBDV subtypes. Phylogenetic analysis showed that 3529/92 isolates belong to the vvIBDV subtype. Recombinant plasmid was constructed and inserted into the yeast expression vector, pPICZ A prior to transformation into Pichia pastoris X33 by electroporation. After the induction of P. pastoris transformant with 0.5% methanol, the production of IBDV polyprotein was observed using Western blot. In P. pastoris, co- or post-translational processing of large polyprotein had occurred generating a stable C-terminal product (VP3).

The consistency of contrast detail performance of imaging system can be evaluated by using Contrast Detail phantom. The analysis is always based on the human visual perception which leads to intraobserver perception bias. In addition, the wall of single drilled hole concept in commercial Contrast Detail phantom gives effect to the penetration of x -ray beam divergence to pass through the base of each hole. This effect will lead to false appearance of image but it is not visualised in the radiograph. In this study, cylindrical double hole acrylic block of Multiparameters Contrast Detail phantom has been developed which differs from the single drilled hole concept, whereby it consists of combination of different holes’ diameters and thicknesses. Results revealed that, the new design of cylindrical double hole acrylic block is able to visualise the effect of image displacement from the x-ray focal length plus measuring the off-position of anode stem, blurring effect, image distortion in terms of real shape and size, also addressing the contrast detail characteristics parameter in terms of real hole depth. The influencing factors of source-image-distance, object thickness, position of object from center beam in x-axis, sizes of hole diameter and hole depth contribute to the changes in parameters’ outcome. The measurement of pixel intensity by using software and development of algorithm for data analysis basically can reduce the human perception bias and increase the validity of the results.

An alternative osteo-healing formulation with osteo-healing properties was formulated by combining gentamicin and Nigella sativa oil (NSO) in a form of gentamicin-N. sativa fusion emulsion (GNFE). This work aims to formulate a stable emulsion and to study the effects of GNFE on UMR-106 osteoblast-like rat osteosarcoma cell line in vitro and its related mechanisms of bone healing and regeneration. Emulsion A, B, C and D had been formulated, with final concentration of gentamicin was made constant at 0.1%, whereas NSO concentration was varied at 32.5%, 35.0%, 40.2% and 46.4% in all formulations respectively. Stability studies of emulsion A, B, C and D were performed at different storage conditions (8°C, 25°C and 50°C), followed by in vitro study of MTT assay, Alizarin Red S (ARS) staining, von Kossa staining and quantification, alkaline phosphatase (ALP) quantification and quantitation of collagen type-1 and osteocalcin (qPCR). Results showed that all emulsions were stable at storage temperature of 8°C. In vitro results showed that emulsion D produced the highest cell viability (97.1%) at 72 hours of post-incubation. The highest mineral deposits (2.64 ± 0.05) and ALP activity (2.19 ± 0.3 nmol) was produced by emulsion D at day 21. Lastly, the highest expression of collagen type-1 (29.4 ± 1.01 folds) and osteocalcin (1.8 ± 0.51 folds) were expressed by the cells treated with emulsion C. Thus, stable GNFE may have the ability to promote bone formation.

The aim of this study is to develop an innovative halal high calorie supplement by incorporating milk, yoghurt, banana, mango and belimbing dayak (Baccaurea angulata) juice. Three formulations with different main ingredients (F1-milk, F2-chocolate-flavoured milk and F3-soy milk) were subjected to sensory evaluation (appearance, odor, taste, mouthfeel and overall liking) to determine its acceptability by using 9-point Hedonic Scale. From the three formulations, the one with the highest Hedonic score was drum-dried for proximate analysis. The drum-dried formulation was stored for 3 months in room temperature to determine its shelf-life by peroxide value determination and microbiological testing. Proximate analyses, chemical and microbial testing were carried out monthly during the 3 months period. All formulations showed fair acceptability. The formulation with the highest hedonic scores was the F3 formulation (6.47 ± 1.77), (p = 0.05) and the calorie content determined was approximately 236.6 kcal/250 ml. The shelf-life study of powdered F3 formulation showed that in a period of 3 months, the peroxide value increased from 0.05 ± 0.00 meq/kg to 0.71 ± 0.00 meq/kg. In addition, there was a decreased in the total plate count (1.25 x 102 ±7.071 cfu/g) and yeast and mould count (<100 cfu/g) at the end of 3 months storage period. From human study, group receiving F3 formulation showed higher weight gain (1.29%) as compared to group that received Enercal ® Plus (0.09%). But the subjects’ Body Mass Index (BMI) values were still in the underweight range. However, there is no significant difference between the two groups (p = 0.25, p>0.05). Thus, through the modification of functional food, it is possible to develop a high calorie supplement with the potential to be a weight gain and high energy supplement.

Cryptosporidiosis, resulting from infection of obligate intracellular protozoan parasite known as Cryptosporidium spp. is a major causative of diarrhoeal diseases in small ruminants. The study aimed to determine the prevalence of Cryptosporidium spp. in relation to age groups and to access the first distribution and genotypes of Cryptosporidium spp. using molecular tools in Malaysian goats from four commercial farms in Terengganu. A total of 454 faecal samples from three age groups (kid, yearling and adult) were collected from selected farms located in Besut, Kuala Terengganu, Marang and Setiu, Malaysia. The samples were subjected to qualitative examination using combination of modified formalin-ether concentration technique (M-FECT) and modified Ziehl-Neelsen (MZN) staining. Cryptosporidium spp. positive samples were withheld for molecular characterization using nested PCR assay targeting the 18S rRNA gene. Of the representative samples, overall prevalence of Cryptosporidium spp. in the present study was 48.7% (221/454). Significant difference (p < 0.05) was observed among age groups, with the goat kids having the highest infection rate (54.6%, 77/141) followed by adult goats (50.2%, 101/201) and yearlings (38.4%, 43/112). The 18S rRNA-based PCR successfully identified the zoonotic species of C. parvum among all age groups. The prevalence study indicated that Cryptosporidium spp. infections are highly distributed among goats in four farms in Terengganu. The genotyping study signified that goats should be considered as one of the most important reservoirs for C. parvum in the studied farms. It is suggested that there is the possibility of zoonotic transmissions of cryptosporidiosis among the goats and animal handlers. In all, the current molecular epidemiological status of cryptosporidiosis in these farms can pose a great economic impact and constraint to the goat industry in Terengganu in achieving better and sustainable productions. Therefore, the need of further molecular subtyping of Cryptosporidium spp. and genotypes together with well-coordinated and extensive sanitary monitoring by animal handlers and field veterinarians are greatly recommended to minimize the occurrence of infections.

Cryptosporidium are coccidian, obligate intracellular parasites that infect human, livestock animal, domestic pet and wildlife worldwide. Protozoal infections particularly by Cryptosporidium are rampant in human and animal leading to health problem and significant economic losses worldwide. Since there were limited data on the prevalence and no molecular characterization of Cryptosporidium especially in goats in Malaysia, this study were aimed to identify the prevalence of Cryptosporidium in goats from three different farm management systems in Terengganu, Malaysia and to determine the species of Cryptosporidium in goats by using 18S rRNA gene. A total of 478 goat faecal samples were collected from three different farm management systems which comprised a total of six farms in Terengganu from February to November 2015. Cryptosporidium oocysts were identified in preserved faecal sample by using formalin- ether concentration technique and observed under the microscope upon staining with modified Ziehl-Neelsen. Cryptosporidium-positive faecal samples confirmed by modified Ziehl-Neelsen staining were kept in 2.5% potassium dichromate prior to DNA extraction. The samples were screened by nested PCR and genotyped by using 18S rRNA gene. The overall prevalence of Cryptosporidium in this study was 43.3% (207/478) and the highest prevalence of Cryptosporidium was recorded in goats from Farm E with the percentage of 56.7% (34/60). There was a significant difference (p<0.05) in the prevalence rate of Cryptosporidium infections in goats among the six farms in Terengganu. Besides, goats reared under the intensive farm management system gave the highest prevalence of infection (49.6%), followed by extensive farm management system (37.1%) and the lowest was semi-intensive farm management system (32.6%). There was a significant difference (p<0.05) between the prevalence of Cryptosporidium and farm management systems in Terengganu. Besides, the Cryptosporidium species identified in goats was Cryptosporidium parvum. Therefore, it is hoped that this study will provide information on the occurrence of Cryptosporidium infection in goats in Malaysia as well as providing better understanding on the public health significance of cryptosporidiosis in order to plan effective control measures.

The treatments of chronic osteomyelitis are difficult, time-consuming and relatively expensive due to the presence of bacterial biofilm that is highly resistant to antibiotics. This study aimed to assess synergistic antibacterial activities of gentamicin-Nigella sativa fusion towards the most common biofilm-bacteria in osteomyelitic infection. Briefly, a total 57 samples (prostheses, bones, tissues and swabs) were taken from 17 cases of osteomyelitic infection at Hospital Tengku Ampuan Afzan, Kuantan. The samples were processed, isolated species were identified, as well as biofilm identification and antibiotic sensitivity assays were performed. Fusion of gentamicin and N.sativa were formulated in 4 different types of emulsions (A, B, C, and D) consisting of constant 0.1% (w/v) gentamicin and different Nigella sativa oil concentrations from 32.5% to 46.6% (v/v). Antimicrobial activities of the emulsions were evaluated using disc diffusion assay and determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Then, the assessment of antibiofilm activities was carried out as pre- and post-biofilm assays. The pre-biofilm consists of biofilm formation inhibition and minimum biofilm inhibition concentration (MBIC). The post-biofilm assay was done to evaluate the effects of the emulsions on the biofilm, using biofilm penetration test and confocal laser screening microscope (CLSM) analysis. It was found that prosthesis (89%) and bone (66.7%) samples produce the most bacteria growth and Staphylococcus aureus (10 out of 16) was the most frequently identified. In the disc diffusion assay, significant synergistic effect of emulsions was seen only in resistant S.aureus (clinical isolate) (Tukey’s test p < 0.05). Additionally, emulsions MIC values were up to 10 times lower than gentamicin alone against all S.aureus while MBC values of emulsions were up to 3 times lower towards sensitive S.aureus (clinical isolate and control). No bactericidal activity was exhibited by all compounds tested on resistant S.aureus (clinical isolate). In pre-biofilm evaluation, there were significant differences in biofilm formation inhibition in comparison between these emulsions with N.sativa and gentamicin alone in both clinical isolate S.aureus (sensitive and resistant) (Tukey’s test p < 0.05). MBIC values of emulsions were up to 10 times lower than gentamicin against all S.aureus. In contrast, N.sativa alone was lesser than emulsions and gentamicin. For post-biofilm assessment, no significant difference in penetration rate was found between emulsions and gentamicin. As opposed to N.sativa which showed little penetration. In the CLSM analysis, only emulsion C was used. Results revealed that emulsion C significantly reduced the biofilm thickness compared to gentamicin and N.sativa alone (Tukey’s test p < 0.05). Furthermore, the surface percentage (%) of non-viable bacteria of emulsions is significantly higher than gentamicin and N.sativa alone (Tukey’s test p < 0.05). In conclusion, this new fusion of gentamicin-N.sativa have synergistic antimicrobial and antibiofilm properties towards different strains of S.aureus including resistant strains, thus, can be developed as a new, and customized, gram-positive-specific treatment for ostoemyelitic infection.

Polyamines are vital in maintaining human health because they perform certain functions that are necessary for cell development. However, extensive intracellular polyamines may promote unwarranted cell proliferation and to a certain extent, stimulates cancer initiation. Therefore, this study aimed to investigate the polyamines deficient diet in chemoprevention strategy using selected fruits recommended by the prophet against human lung adenocarcinoma cells, A549. Five prophetic fruits were selected, including Phoenix dactylifera (ajwa dates), Beta vulgaris (beetroot), Ficus auriculata (fig), Ziziphus jujube (jujube) and Vitis vinifera (raisins). Initially, they were freeze dried and stored in -80°C until analysis. The polyamines in each selected prophetic fruit was quantified and classified by High Performance Liquid Chromatography (HPLC). Subsequently, their anti-proliferative effect on A549 cells growth was evaluated using MTT assay and Trypan blue exclusion assay. Protein content was measured using Lowry assay. The activity of genes that regulate polyamine metabolic pathway particularly ornithine decarboxylase (ODC) and spermine/spermidine (N1)-acetyltransferase (SSAT) was elucidated using qPCR. Finally, cell cycle profile and apoptosis assay were conducted with flow cytometer, while caspase assay was done using colorimetric method. It has been found that jujube showed the highest polyamines concentration (219.6 ±4.4 nmoles/ml) while fig was the lowest (39.3±3.0 nmoles/ml). MTT assay suggested IC50 of fruits recommended by the prophet was ranged from 15 mg/ml to 30 mg/ml. All selected prophetic fruits showed anti-proliferative effect against A549 cells. Protein content was significantly lower after 72 h of treatment with selected prophetic fruits, and total elimination of intracellular spermidine and spermine were observed in all treated A549 cells. The polyamines metabolism was found to be altered in which the intracellular polyamines depletion was mainly contributed by the downregulation of ODC in A549 cells treated with ajwa dates and jujube. Meanwhile, beetroot and fig induced the upregulation of SSAT. Cell cycle profile displayed significant cell cycle arrest at G2/M after 48 h of exposure to jujube and raisins. It is demonstrated that beetroot, fig, jujube and raisins induced apoptosis after 48 h exposure while ajwa dates might induced other type of cell death. Caspase assay revealed significant activation of caspase 3, 8 and 9 in beetroot treated cells while no caspase activation was identified in other prophetic fruits treated cells. Considering the classification of polyamines, the anti-proliferative effect and the type of cell death, it was concluded that fig, raisins and beetroot are the promising candidates for nutritional cancer therapy and preventive approaches for cancer.

Cancer is a complex disease to treat and has poor survival rate. Natural product has shown a promising effect in cancer treatments. Punica granatum or pomegranate has a potential to be a useful cancer preventive agent, however the mechanism of action is unclear. To understand how effective the pomegranate in preventing cancer growth, it is crucial to identify the signaling pathways affected. One possibility is the polyamine pathway which is important in many cells function and its up-regulation of this pathway in cancer makes it a logical target for cancer prevention. Therefore, this study was aimed to evaluate the role of pomegranate and polyamines in regulating cell cycle distribution and cell death mechanisms as well as its anti-proliferative effect in human lung adenocarcinoma cells, A549. The classification of polyamines in pomegranate juice was determined by HPLC analysis. The anti-proliferative effect of pomegranate juice was tested by using MTT assay and the effect of 2% pomegranate juice on A549 cells growth and viability were evaluated by trypan blue exclusion assay. The cellular protein and polyamines content in A549 cells were determined using Lowry assay and HPLC, respectively. The cell cycle distribution and cell death mechanisms were evaluated using flow cytometer. The gene expression was evaluated using quantitative real - time PCR. The study found that pomegranate juice was classified under low polyamine diet. At the concentration range from 0 to 3%, pomegranate juice caused inhibition of A549 cells growth. The inhibitory concentration (IC50) was 1.4% ± 0.23, 1.5% ± 0.14 and 1.4% ± 0.12 after 48, 72 and 96 h exposure, respectively. At the concentration of 2%, inhibition of growth was observed by showing decreased in viable A549 cell number and protein content after 48 h (p<0.05), 72 h (p<0.001) and 96 h (p<0.001) exposure compared to untreated A549 cells. In contrast, the percentage of cells viability remains high and constant. The results also showed a positive correlation between total viable A549 cells number with protein content where the R2 values for untreated and treated were 0.9248 and 0.6523. There were no significant differences in total polyamines content in untreated and treated A549 cells. The study also found that pomegranate juice induced cell cycle arrest at G0/G1 phase and apoptosis via intrinsic pathway following 24 h treatment. Pomegranate juice caused loss of mitochondrial membrane permeability after 48 h (p<0.05) exposure and a release of cytochrome c in cytosol after 24 h (p<0.05) and 48 h (p<0.01) exposure in treated A549 cells. In caspases analysis, it was showed that there was activation of caspase-3 following 72 h (p<0.01) treatment and caspase-9 after 48 (p<0.01) and 72 h (p<0.05) exposure in treated A549 cells. Lastly, gene expression study found that pomegranate juice inhibited the expression of ODC gene after 24 h and 48 h exposure (p<0.001) and SSAT gene after 48 h exposure (p<0.05) in treated A549 cells. From the study, it can be deduced that the suppression of A549 cell growth might not due to modulation of polyamine metabolism. However, pomegranate juice able to cause A549 cell growth inhibition by inducing cell cycle arrest and apoptosis through mitochondrial pathway.