Road traffic accidents are increasingly recognised as a serious public health concern. Each year road traffic accidents cause 1.2 million deaths worldwide and the number is expected to increase by 65 percent in 20 years. One of the challenges to solving this issue is that many accident investigation studies employed methods that gave less emphasis on systemic factors of road traffic accidents such as enforcement and government policies but rely on a direct cause-effect chain to analyse an accident causation process. By contrast, more contemporary approaches of investigating accidents are based on systemic accident models that can be used to analyse the interactions between various components of a sociotechnical system. Different systemic accident models, however, can lead to different analyses and subsequently different conclusions. As such, this study sought to compare three different systemic accident models, namely, the Driving Reliability and Error Analysis Method (DREAM), System-Theoretic Accident Model and Process (STAMP), and AcciMap in analysing the accident causation process of the 2013 Genting Highlands Bus Crash, which was the deadliest road traffic accident in Malaysia. Using DREAM, this study found that most of the contributing factors originated from maintenance errors and road design issues. On the other hand, STAMP identified inadequate controls and flaws within the road transportation system as contributing factors while AcciMap indicated various direct and indirect factors across different levels within the system. Findings of this study suggest that DREAM can present the accident factors in a succinct graphical representation by using a classification system thus making it easy to carry out and simple to understand. In addition, although both AcciMap and STAMP analyse road traffic accident across different system levels, STAMP is more comprehensive and flexible. However, because STAMP is time consuming and complex, it is more suitable for analyzing major road traffic accidents.

Holothuria scabra is one of edible marine animals which is widely consumed by Chinese and local Sabahan people, who believe in the medicinal properties offered by the species. This work was focusing on H. scabra in the attempt of finding new therapeutic properties, because of its large availability in the coastal area of Sabah and ability to be widely bred in hatcheries for restocking purpose. The study comprises of two types of study: anti-gastric ulcer and acute toxicity test, to investigate the existence of anti-gastric ulcer property in the species and its safety for human consumption. In the anti-gastric ulcer study, 150 mg/ml H. scabra water extract was administered into rats that had been induced with gastric ulcer by using acetic acid. Histology of the stomach was examined and the gastric mucous was estimated by using spectrophotometer. The toxicity test was conducted by giving H. scabra water extract with the doses of 5, 50, 300 and 2000 mg/kg into four groups of rats, whereas the control group received distilled water. Survival and toxicity signs were carefully observed for 14 days, in addition to performing the blood analyses and histopathological examination of liver and kidney tissues. For the anti-gastric ulcer study, the stomach tissues from both groups of treated and untreated were recovering, with no significant difference in terms of structural changes and inflammatory cell infiltrates. The treated rats did not produce significant higher amount of gastric mucous as compared to the non-treated rats, except on the day 12 of the experiment. In the toxicity test, no mortality and toxicity signs were observed throughout the observation period. Gross examination on liver and kidneys showed no changes in appearance, size, and colour of the organs when compared to the control group. Statistical analysis of the Kruskal-Wallis H test produced no significant difference in body weight between the control and all treated groups after the treatment, x2(4, N = 23) = 5.754, p = 0.218. The haematological and biochemical blood parameters of the treated group of rats also showed no statistical difference as compared to the control. However, histopathological examination revealed presence of necrotic tissues and cellular damages in the liver and kidneys of all treated rats at all doses of extract. The study suggests that the effects of H. scabra water extract on the healing of induced gastric ulcer in rats were not conclusive. However, since blood analyses results and histopathological findings of the acute toxicity study were contradict, sub-acute toxicity is suggested to be conducted in future to further confirm the toxicity level of the extract.

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.

Newcastle disease is a very contagious disease in chickens and one of the most important diseases of poultry in the world. This study aimed to compare the effect on immune response and virus shedding in vaccinated chickens of the new inactivated vaccine of Newcastle disease virus (NDV) and commercial vaccine LaSota. The local subtype of NDV Genotype VIId (isolate 12234/10) was chosen as inactivated vaccine candidate and three different adjuvants were used which includes Oil-emulsion Complete Freund’s, Oil-emulsion Squalene-based, and aluminium hydroxide gel (Alum). Ten different groups of chicken consists of ten each, were vaccinated with and without adjuvant of the inactivated vaccine via intramuscular and subcutaneous, respectively. As a comparison, a group of chicken was vaccinated with commercial vaccine and control group was not vaccinated. Then, all of the chickens were challenged with NDV Genotype VIIh (isolate 7979/01) two weeks after vaccination. Blood sera of the chickens were taken weekly for haemaglutination-inhibition (HI) test starting from prior to vaccination. Virus shedding of the chickens were calculated based on the virus isolation done within two weeks after challenge. With 100% survival rate and highest mean HI titre of log2 7.8, the inactivated vaccine with adjuvant Oil-emulsion Complete Freund’s surpassed the LaSota and the virus shedding also reduced significantly. Alum-adjuvanted inactivated vaccine gave most impressive results as LaSota with all chickens survived at log2 6 of antibody titer and zero virus shedding. Whereas, without adjuvant, the chicken HI antibody titre is below log2 4 after vaccination and only 20-30% were survived. Based on the post-mortem findings on the survived chicken from each vaccinated group, their internal organs appeared normal and no sign of haemorrhage or pathognomonic signs of Newcastle disease (ND). Conclusively, vaccinated chicken are effectively protected from morbidity and mortality against virulent genotype VII challenge with the addition of adjuvant into inactivated local strain of NDV genotype VII vaccine. Thus, the development of inactivated local NDV genotype VII vaccine is a promising candidate to control the current ND endemic in Malaysia.

Breast cancer is a leading cause of cancer-related death in women. Despite the advancement of breast cancer treatments, patients are suffered with side effects and other consequences such as cancer recurrence and resistance to treatments. An effective strategies are needed of which resources from the natural plants have been used for alternative approach as anti-cancer agents against breast cancer. The present study is an attempt to evaluate anti-cancer activities of Neolamarckia cadamba leaf (NCL) extract on the breast cancer carcinoma cells (MCF-7). MCF-7 cells were treated with various concentration of ethanolic NCL extracts. The concentration of NCL extract that caused 50% of cell growth inhibition [IC50] in MCF-7 cells was determined after 72 hours of treatments, and its anti-proliferative effect was assessed upon treatment for 24, 48, 72 and 96 hours using TBEA method. NCL extract demonstrated inhibitory activity in cell growth at IC50 value of 0.206 mg/ml, in dose-dependent and time-independent manner. The effect of NCL extract on apoptosis induction and cell cycle arrest were analysed using flow cytometer. The flow cytometric analysis indicated that NCL extract inhibits the growth of MCF-7 cells by inducing apoptosis and cell cycle arrest in G0/G1 phase. Meanwhile, gene expression analysis of qPCR assay revealed NCL extract induced apoptosis through the down-regulated of Bcl-2, whereas Bax cytochrome c, caspase-9 and caspase-7 were up-regulated in MCF-7 cells. Cell cycle arrest was associated with down-regulated of CDK2 with subsequent up-regulated of p21 and cyclin E. Overall, our data demonstrated that the NCL extract exerts anti-cancer effect on MCF-7 human breast cancer cells through induction of apoptosis and cell cycle arrest. The present study suggests that NCL extract may be an important alternative of anti-cancer agent candidate in breast cancer treatment.

Breast and lung cancer have highest mortality rate among female and male respectively. Even though with the availability of advanced treatment, the mortality rate is still increasing. Thus, this study aims to elucidate the potential Malayan porcupine bezoar (PB) in exhibiting anticancer effects on the breast (MCF7) and lung (A549) cancer cells. Porcupine Bezoar extracts were tested with optimized IC50 for its potential ability in inhibiting cell growth, changing morphology, inducing apoptosis and cell arrest. The PB extract showed the growth inhibitory effect with IC50 value at 19.0 µg/ml (MCF7) and 13.5 µg/ml (A549) respectively. The morphology assessment upon PB extract treatment at 72 hours displayed possible apoptosis features of pyknosis and karyorrhexis. Further investigation with Annexin-V/7AAD revealed PB treated cell induced early and late apoptosis with a presence of dead cells in 72 hours. Further test has shown, PB may have altered cell cycle regulation and arrest in G1 for both cells. Possible signaling pathway at a molecular level for both cells was investigated. The result revealed induction of apoptosis following intrinsic pathway by initiating the release of Cytochrome C from mitochondria thereby activating caspase cascade. The above claims were supported by downregulation of Bcl-2 which suggest PB extracts triggered apoptosis. Furthermore, the investigation was conducted on cell cycle regulator genes, revealed the up-regulation of p21 and down-regulation of cyclin D suggesting cell arrest in G1 phase. Additionally, finding demonstrate PB extracts was selective in inducing significant effect on cancer cells MCF7 and A549 compared to normal fibroblast 3T3-L1 and HGF-1. This research report unveil the potential medicinal properties of porcupine bezoar in anticancer perspectives.

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 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.

Monopterus albus is one of the terrestrial animal sources that is widely consumed by the local people, who believe in the medicinal properties offered by the species. However, there is not many research has been done on the species to prove the claims. Therefore, this study was conducted to evaluate the potential of M. albus extracts as an aphrodisiac and to characterize the components of the extracts. This study comprises of three phases of study: aphrodisiac, fertility and acute toxicity test. Two types of extraction were used; aqueous and lipid. Prior to treatment, the extracts were analysed for the mineral contents and fatty acid analysis using inductively coupled plasma optical emission spectrometry (ICP-OES) and Gas Chromatography Mass Spectrometry (GC-MS), respectively. Then, the sexual behaviour was observed in eight groups of ICR male mice with four animals in each group (n = 4). The doses used for both extracts were 50, 100 and 200 mg/kg. Negative control group received 0.9% saline, meanwhile positive control group received 5 mg/kg Sildenafil citrate or known as Viagra®. After an hour administration (i.p.) of the respective doses, the observation of sexual behaviour was conducted. The male mouse was paired with a female mouse for three hours to determine the effect of extracts on sexual behaviour. In addition, mating assessment also was carried out at similar dose level. On the sixth day, the copulatory plug was observed on the female mice used to confirm the presence of ejaculation. Afterwards, the male mice were slaughtered to evaluate the sperm count and motility as well as testicular histology. Another group of male mice were used for the analysis of testosterone levels. After that, toxicity test was conducted to determine the effect of a single dose of lipid extract in rats. The rats administered orally with the doses of 5, 50, 300 and 2000 mg/kg of M. albus lipid extract and control group received 0.9% saline. The survival and toxicity signs were observed for 14 days. Besides that, the blood analysis and histopathological examination of liver and kidney tissues also were investigated. From the analysis of free fatty acid, it shows that lipid extracts of M. albus contained higher level of arachidonic acid (AA), the essential fatty acid which influences the reproduction. The results obtained shows that all doses of lipid extract has significantly increased (p ≤ 0.05) the mounting frequency (MF) and sperm count as compared to negative control. Meanwhile, in aqueous extract, only the dose of 200 mg/kg shows significant difference of MF but no improvement in sperm count. In the analysis of testosterone levels, only the dose of 200 mg/kg lipid extract and positive control group showed tendency to increase the hormone levels in treated mice. Whereas, in the toxicity study, the haematological, biochemical and histological analyses showed no significant difference (p ≥ 0.05) as compared to the control. The treated rats showed no acute signs of toxicity and no mortality was observed. Therefore, the M. albus was safe to be consumed by the people and has great potential as an aphrodisiac agent. Thus, the study suggests that M. albus lipid extract could be used as an alternative source for sexual activity in men.

Despite the enforcement of Environmental Quality Act in 1974, water quality of rivers in Malaysia is still deteriorating. Information collected regarding water quality in rivers especially in the state of Pahang is still insufficient. Therefore, the objectives of this study were to detect the occurrence of Cryptosporidium oocysts and to assess the physico-chemical and heavy metal parameters in Kuantan River and Balok River. Six water samples were collected from Kuantan River and Balok River. Water samples were filtered followed by the purification of Cryptosporidium oocyst using the immunomagnetic separation technique and stained by fluorescein isothiocyanate–staining. Physical parameters were measured directly at the sampling locations using the handheld multi-parameter instrument while the assessment of chemical and heavy metal were done in the laboratory. Results obtained were then compared with the Interim National Water Quality Standards (INWQS) which is the standard parameters used to assess the water quality in surface water in Malaysia. Kuantan River and Balok River were contaminated with Cryptosporidium oocysts and chemical contaminants. However, water quality in both Kuantan River and Balok River varies depending on the sampling points. Cryptosporidium oocysts were detected in all water samples. With regards to INWQS, our results reported a lower level of dissolved oxygen (<5 mg/L) and elevated level of chemical oxygen demand (>50 mg/L), biological oxygen demand (>4 mg/L), nitrite (>1 mg/L) and chloride (>200 mg/L) at certain sampling locations. Of 23 elements measured, only 9 elements were detected (aluminium, boron, calcium, cobalt, iron, potassium, magnesium, sodium and strontium). The presence of heavy metals; aluminium (>0.5 mg/L) and iron (>0.8 mg/L) at several locations were beyond the INWQS safety limit range. Results obtained from this study will be very useful for future river management. Further studies on molecular characterization of Cryptosporidium species in river is highly recommended. Besides that, more parameters can be added in the future for more extensive information. It is hoped that this study can provide new data to the growing body of literature on the presence of Cryptosporidium oocyst as well as the physico-chemical assessment in Kuantan River and Balok River.

Rare earth elements (REEs) are a series of chemical elements found in the earth’s crust that are vital to many modern technologies, including consumer electronics, computers, communications and clean energy technologies. However, during the production of REEs, the activity is susceptible in causing environmental contamination by discharging the REE industrial wastewater into the environment especially ponds, lakes and rivers. The REE wastewater containing Lanthanum (La) residue and other radioactive elements will be uptaken by the aquatic plants and freshwater fishs through bioaccumulation process. Therefore, this study aims to determine the effectiveness of the aquatic plant, i.e. Pistia stratiotes and freshwater fish (Oreochromis niloticus) as bioaccumulation agents of rare earth metals, especially Lanthanum (La) in REE artificial wastewater system. The study was carried out by subjecting P. stratiotes (water lettuce) and O. niloticus (Nile tilapia) to 4 levels of Lanthanum Chloride (LaCl3) treatments (i.e. 1, 3, 5 and 10 ppm) for 12 days in aquaponics artificial wastewater system. The samples were collected every two days and dried prior to digestion. The dried plant parts (root and leaf) and fish tissues (stomach and flesh) were acid-digested and then analyzed the REE metal (La) accumulation using the Inductively Coupled Plasma-Mass Spectrometry (ICP-MS). Overall results showed REE (i.e. La) was accumulated in the highest amount in the stomach compared to other parts of the plant and fish tissues.

The addition of fruits fiber in bakery products such as biscuit has been shown to improve blood glucose response. Bacaurea angulata is one of the underutilized fruit, which may have some potential benefits to health. The effect of two different ways of drying, which are hot air drying and freeze drying on chemical properties of B.angulata was examined. The chemical analysis includes proximate analysis, antioxidant properties and dietary fiber composition. Four different amount of hot air dried B.angulata pomace (0 %, 10 %, 20 % and 30 %) were incorporated in the biscuit formulations substituting wheat flour. Shortening content was reduced to 40 %, 30 % and 20 % of the weight flour. The effects of supplementation of hot air dried B.angulata pomace on physical qualities as well as sensory properties of sixteen biscuits were determined. Four formulations with the highest overall acceptance score were selected for further analysis on chemical properties and blood glucose response. There were full fat control biscuit (FFC), full fat with 10 % pomace biscuit (FF10), full fat with 20 % pomace biscuit (FF20) and 40 % fat with 10 % pomace biscuit (RF10). Blood glucose response was assessed on twenty healthy non-diabetic subjects, where they were required to consume reference food (glucose drink) and four test biscuits after an overnight fasting in different sessions with at least one day gap. Each test meals contained 50g available carbohydrate. Blood samples were collected from finger-pricks in fasted subjects before each test (0 min) and at 15, 30, 45, 60, 90 and 120 minutes after consumption of reference or test biscuits. The GI was calculated as ratio of incremental areas under curve (IAUC) of test biscuits with IAUC of reference food. Hot air dried pomace contained significantly higher moisture and lignin in comparison to the freeze-dried pomace. In contrast, freeze-dried pomace showed significantly higher total ash, cellulose and resistant starch. There were no significant differences in total, insoluble, soluble dietary fiber and total fructan content with different types of drying. Freeze-dried pomace also sustained better antioxidant activity showed by significantly lower IC50 value of DPPH and ABTS, 39.78±0.69 mg/ml and 4.98±0.39 mg/ml respectively in comparison to the hot air dried pomace. The physical and sensory properties of pomace fortified biscuits were significantly affected by fat and pomace level, where FF10 biscuits showed the best preferences among consumers. However, further chemical studies suggested greater nutritional values in FF20 biscuit, where it contained the highest total dietary fiber (6.65±2.90 g/100g) and resistant starch (0.4±0.14 g/100g) although not significantly different from FFC biscuit. FF20 biscuit also contained significantly higher antioxidant properties, DPPH (0.76±0.29 mg TE/g) and ABTS (3.69±0.15 mg TE/g) in comparison to other type of biscuits. All four tested biscuit types, including the FFC biscuit, showed low GI ranging from 44 to 54. Therefore, fortification of biscuits with B.angulata pomace was suitable to produce low GI food.

Breast cancer is a major health challenges in worldwide including Malaysia. It is the commonest malignancy among women. Although there are several of cancer treatments; however, these current treatments have consequences to the cancer patient such as recurrence of cancer, drug resistance and severe side effects. Discovery of alternative treatment which have minimal or almost no side effect is needed for cancer treatment. Traditional practitioners in India used the powder form of Luffa aegyptiaca seeds (LAS) in treating breast cancer. The old folks in Malaysia also used the powder form of LAS to treat the breast cancer patient. Thus, the present study is aim to determine the chemopreventive effect of LAS extract on breast carcinoma cell (MCF-7) by determining IC50 value, antiproliferative effect, apoptosis induction and cell cycle arrest via flow cytometric analysis as well as molecular signalling pathway of related genes expression with regard to apoptosis induction and cell cycle arrest by using qPCR analysis. MCF-7 cells were treated with various concentrations of ethanol extract of LAS. IC50 value of 80 μg/mL was determined after 72h of LAS treatment on MCF-7 cells and its anti-proliferative effect was assessed for 24, 48 and 96 hours using TBEA method. LAS extract demonstrated inhibitory activity in cell growth in a dose-dependent and time-dependent manner. The effect of LAS extract on apoptosis induction and cell cycle arrest indicated that it inhibits the growth of MCF-7 cells by inducing apoptosis and cell cycle arrest in G0/G1 phase. Meanwhile, gene expression analysis of qPCR assay revealed that LAS extract induced apoptosis through the down-regulation of Bcl-2, whereas Bax cytochrome c, caspase-9 and caspase-7 were up-regulated in MCF-7 cells. Cell cycle arrest was associated with down-regulation of CDK2 with subsequent up-regulation of p21 and cyclin E. Overall, the research finding provide new insight that Luffa aegyptiaca seeds (LAS) is a promising candidate of chemopreventive agents on MCF-7 cells.

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.

Cervical and ovarian cancers are the lethal gynaecological malignancies that affect the local health scene among women. Eurycoma longifolia, Nigella sativa and Hibiscus sabdariffa are widely used as herbal remedies and natural supplements in Malaysia. The current study is designed to investigate the cytotoxic effects of standardised quassinoid E. longifolia (TAF 273), N. sativa (NSE) and H. sabdariffa (HSE) extracts on cervical cancer (HeLa) and ovarian cancer (Caov-3) cells and the mechanism of cell death. The cytotoxicity of TAF 273, NSE, and HSE were evaluated using MTT assay and the mode of cell death was detected by Hoechst 33258 nuclear staining. The analysis of apoptosis and cell cycle arrest were assessed via flow cytometry. The gene expression of Bax, Bcl-2 and caspase-3 were investigated through real-time PCR. The findings showed that TAF 273 suppressed the cell growth of Caov-3 with the lowest IC50 value (5.3µg/mL), comparable to the cisplatin (IC50 value of 6.07 ± 1.87 µM; ~6.1 µM). The IC50 values of TAF 273 on HeLa as well as NSE and HSE on both Caov-3 and HeLa were however not identified. TAF 273 and cisplatin showed no toxicity on normal human ovarian surface epithelial cells (nHOSEpiC). The characteristics of apoptosis such as cell shrinkage, chromatin condensation and nuclei fragmentation were found following TAF 273 and cisplatin treatment. The flow cytometry analyses showed that TAF 273 and cisplatin induced the apoptosis on Caov-3 in dose- and time-dependent manner and arrested the cell cycle at G2 phase. The real-time PCR results showed that TAF 273 down-regulated the Bax, Bcl-2, and caspase-3 expression. Meanwhile, cisplatin up-regulated the Bax and caspase-3 and down-regulated the Bcl-2 expression. Results from this study suggests that TAF 273 was cytotoxic on Caov-3 cells through the induction of the apoptosis via the mitochondrial pathway, initiated by proteins other than Bax and it was executed through other group of executioner caspase or via caspase- independent pathway. The output of this study may indicate the potential of the standardised quassinoid of Eurycoma longifolia extract in the successful treatment of ovarian cancer as part of cancer therapy.

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.

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.

Elderly population is at risk for malnutrition due to various factors. To address this issue, the elderly needs to be screened for malnutrition and received an appropriate care plan if necessary. The use of nutrition resource is useful as part of a care plan due to its ability to provide nutrition education for malnourished and at-risk elderly. Therefore, this thesis aimed to develop and evaluate a nutrition resource kit for nutrition intervention among malnourished and at-risk elderly in health clinic setting. This multiphase, action research study was conducted in four selected health clinics in Kuantan, Pahang. A mixed-method approach was taken to achieve the research objective: (Phase I: Plan) qualitatively assessed the types of nutrition resource kit needed by community living elderly; (Phase II: Act) development of nutrition resource kit and educational video for malnourished and at-risk elderly and (Phase III: Observe and Reflect) qualitatively explored the acceptability towards the newly developed nutrition resource kit and educational video and improvisation of it. Phase I aimed to demonstrate the types of nutrition resource kit needed by community living elderly in health clinic setting. Twenty-one elderly participants with a mean age of 67.24 ± 6.98 years were purposively recruited for individual in-depth interview. Thematic analysis of the interview identified five themes; 1) provision of nutrition resource kit, 2) preference needed to solve this issue (Webb et al., 2018). Favourable outcomes for printed version, 3) eye-friendly, 4) facilitate understanding with diagram and 5) support for technology-based materials. In phase II, a nutrition resource kit with incorporation of an educational video in a QR code were developed based on the integration of findings in phase I and the scoping review which had been performed to identify the suitable topics to be included. This newly developed kit and video then had undergone validation by experts and elderly. Six experts involved in content and face validation of the kit and video. Both resources achieved excellent content validity. Suggestions by experts were considered and revision was done accordingly. The revised kit and video was further assessed by twelve elderly for face validation. All elderly positively evaluated both resources. Amendment was done according to the elderly’s suggestions for further improvement. Then, assessment on acceptability towards the developed kit and video among malnourished and at-risk elderly was conducted in phase III. Twenty-two elderly participants (mean age = 68.82 ± 5.33 years) were purposively selected for individual in-depth interview and triangulation was performed using a questionnaire. Interviews were thematically analysed and descriptive statistic was used for data triangulation. Four themes emerged; 1) positive acceptance, 2) elderly-friendly, 3) valuable and 4) individual preference. Additional suggestions given by elderly were reviewed and revised accordingly. To conclude, this newly developed nutrition resource kit both in the printed format and electronically was well-accepted by the malnourished and at-risk elderly. Provision of nutrition resource kit tailored to the need of elderly could help to deliver nutritional guidance effectively and serve as an important reference for them to prevent and overcome malnutrition.

Elderly population are susceptible to malnutrition. However, malnutrition identification among community-living elderly through nutrition screening is not routinely performed in Malaysia health clinics; although it is recommended to be performed routinely in all healthcare setting. Major issue to this is due to no specific nutrition screening guideline and validated tool in this setting. Due to this gap, development of nutrition screening guideline is essential for use among healthcare staff. The aimed of this study are to develop a nutrition screening guideline for use by healthcare staff in health clinic setting and assess its feasibility. This study has taken a mixed-method approach and action research to address the research objectives. This study was divided into three phases; Phase 1: Identification of barriers and opportunities of nutrition screening in elderly patients in the health clinic setting; Phase 2: Development of nutrition screening guideline by using Mini Nutritional Assessment Short-Form (MNA® -SF) for healthcare staff in health clinic setting; and Phase 3: To assess the feasibility of newly developed nutrition screening guideline for health clinic setting among healthcare staff. Four health clinics from urban and rural areas in Kuantan, Pahang involved in this study. In Phase 1, in-depth interviews were conducted among twenty healthcare staff. Non-participant observations that acted as triangulation were conducted among twenty-one elderly patients. Both data from interviews and observations were analysed thematically using NVivo software version 12. Four themes emerged for barriers and opportunities were Theme 1: Time; Theme 2: Patient factors; Theme 3: Organization factors and; Theme 4: Nutrition screening knowledge. Findings from Phase 1 has become guidance in content development of the nutrition screening guideline. In Phase 2, twenty-three documents have been included from conducted scoping review for the content development of the guideline. Following this, A4-sized guideline has been developed. The guideline has been validated among six experts by using Item-Content Validity Index (I-CVI). Items that have achieved ≥0.83 were remained. Meanwhile, items with a score <0.83 were revised accordingly. Next, face validity among twelve healthcare staff has been conducted and all participants showed positive responses. Amendment has been made based on provided suggestions. For Phase 3, 22 healthcare staff from similar health clinics participated and had screened five elderly patients each (total=110, mean age= 68.7±6.1 years). After completing nutrition screening, individual in-depth interviews were conducted with the healthcare staff. All data were analysed thematically. The themes obtained were Theme 1: Ease of use; Theme 2: Identification and management of malnutrition; Theme 3: Acceptability; and Theme 4: Implementation of nutrition screening. In summary, this multiphase study has identified appropriate approaches in implementing nutrition screening among elderly patients in health clinic setting. Provision of a comprehensive newly developed nutrition screening guideline tailored to this setting was well-accepted among healthcare staff and feasible in health clinic setting. This nutrition screening guideline is recommended to be adopted nationwide in Ministry of Health’s health clinics towards optimum nutritional status of elderly population and achieving healthy ageing; whilst policy related to nutrition screening implementation in health clinics can be further enforced.

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.