Advances in the treatment of cancer have allowed adolescent patients to become long-term survivors able to lead normal lives. However, the concern about the effects of anticancer drugs on fertility has led to many efforts to preserve germ cells of these patients. This study focuses on ways to assess the effectiveness of Nigella sativa and its active compound, thymoquinone, in lowering chemotherapeutic-associated toxicity of cyclophosphamide on the ovaries and testes of Balb/c mice. Histological and morphological effects on the germ cells were examined via microscopy techniques and possible fragmentations of sperm DNA was assessed using the COMET Assay. The RNA expression of OGG1 and FGF2 in the testicular cells was quantitatively analysed using the real-time PCR followed by quantification of the stages of fertilisation and embryo division using the inverted microscope. Supplementation of N. Sativa oil and thymoquinone exhibited noticeable protective effects on the histology and morphometry of the ovaries and seminiferous tubules as well as being effective in reducing the total DNA fragmentation in spermatozoa. N. Sativa extract reduced the percentage of abnormal sperm head post to cyclophosphamide treatment and preserved the normal chromatin condensation indicative of protection against sperm DNA alteration. The expressions of DNA repair and fibroblast growth factor genes were also shown to increase suggestive of possible reduction in mutagenic modifications. Thymoquinone supplementation increased the implantation and fertilisation rates, pregnancy outcome as well as preserved fair quality embryos following paternal and maternal exposures to cyclophosphamide. N. Sativa and thymoquinone are both suitable exogenous agents that offer viable chemoprotective potential against toxicity induced by cyclophosphamide. This study is part of an effort towards improving interventions to preserve fertility and to assist in the development of techniques in achieving favourable reproductive outcomes for adults who survived childhood cancer following chemotherapy.

Cardiovascular diseases remain the single leading cause of deaths globally. Diet rich in fruits and vegetables is highly important in the maintenance of best possible cardiovascular health. Malaysia has a variety and large quantity of underutilized fruits which are rich in phenolic compounds. In the present study, cardiovascular protective effects of an underutilized Baccaurea angulata fruit were studied using a cholesterol-fed rabbit model. The study was divided into 5 parts. The objective of part one was to identify and quantify some phenolic compounds in the methanolic extracts of B. angulata whole fruit (WF), skin (SK) and pulp (PL). By using the ultra high-performance liquid chromatography-tandem mass spectrometry (UHPLC/MS-MS) based approach, a total of 17 compounds were detected and characterized on the basis of their chromatographic retention time, UV-vis spectra and mass spectra in the negative-ion mode and data from the literature. Part two was aimed at evaluating the effects of WF, SK and PL juices of B. angulata administered at a nutritional dose of 1 ml/kg/day on malondialdehyde (MDA) levels and antioxidant enzymes such as superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT) as well as on total antioxidant capacity (TAC) in rabbits fed with high-cholesterol diet for 12 weeks. The results showed that B. angulata fruit had modulating effects on lipid peroxidation and antioxidant enzyme activities in rabbits fed with high-cholesterol diet. It significantly (p<0.05) lowered the concentrations of plasma MDA. SOD, GPX, CAT and TAC decreased with cholesterol feeding and recovered with B. angulata juice administrations. The purpose of part three was to evaluate and compare the effects of three different B. angulata WF juice doses administered at nutritional doses of 0.5 ml/kg/day, 1 ml/kg/day and 1.5 ml/kg/day on serum, aorta & liver MDA levels, antioxidant enzymes (SOD, GPx and CAT) as well as on total antioxidant capacity in rabbits fed with cholesterol diet for 12 weeks. The results showed that higher doses (1 ml/kg/day and 1.5 ml/kg/day) of B. angulata WF juice exerted better antioxidant activity. Part four of the study was aimed at evaluating and comparing the effects of three different B. angulata WF juice concentrations administered at nutritional doses of 0.5 ml/kg/day, 1 ml/kg/day and 1.5 ml/kg/day on 12 serum, aorta & liver inflammatory biomarkers of atherosclerosis such as vascular cell adhesion molecule 1 (VCAM-1), intercellular cell adhesion molecule 1 (ICAM-1), P-selectin, C-reactive protein (CRP), IL-1, IL-2, IL-6, IL-7, IL-8, IL-10, IL-18, and tumor necrosis factor α (TNF- α) in rabbits fed with cholesterol diet for 12 weeks. The results showed that B. angulata inhibited inflammatory biomarkers of atherosclerosis. Lastly, the aim of part five was to investigate the plaque-reducing activities of B. angulata fruit as a new anti-atherogenic plant in high-cholesterol fed rabbits for 12 weeks. Rabbits fed with cholesterol diet only successfully developed severe, extensive and more densely distributed human-like atherosclerotic lesions. However, supplementation of B. angulata fruit juice to rabbits fed with cholesterol diet significantly (p<0.001) reduced atherosclerotic lesions. In conclusion, B. angulata fruit was shown to enhance plasma antioxidant enzyme activities, inhibit inflammatory biomarkers and reduce progression of atherosclerotic plaque in cholesterol-fed rabbits. Thus, B. angulata fruit plays a favorable role on several mechanisms involved in combating atherosclerosis.

Atherosclerosis is the underlying disease process in the blood vessels that usually results in coronary heart disease (CHD) and cerebrovascular disease, which are the most common forms of cardiovascular disease (CVD). A high level of serum cholesterol is mainly identified as an important risk factor in the development and progression of premature atherosclerosis. Previous studies on Baccaurea angulata (BA), a Malaysian underutilized fruit, showed that it is very rich in fiber and antioxidants. The present study was thus undertaken to evaluate the potential health benefits of BA fruit juice on cardiovascular disease biomarkers in diet-induced atherosclerotic rabbits. The studies were carried out in three distinct phases. In phase one, the effect of solvents [methanol and phosphate buffered saline (PBS)] using cold extraction was examined on the total phenolic content (TPC), total flavonoid content (TFC), and total carotene content (TCC), free radical scavenging activities and lipid peroxidation inhibition activities of the various fruit parts (skin, pulp and whole fruit) using spectrophotometer. The results indicated that the methanol crude extracts significantly (p<0.01) contained higher TPC, TFC and TCC than PBS extracts in all the fruits parts. The edible portion had the highest and most significant (p<0.01) TPC (15357.77±150.72 µg gallic acid equivalence (GAE)/g), TFC (37.32±0.55 mg quercetin equivalence (QE)/g), and TCC (6571.43±185.86 µg ?-carotene equivalence (BC)/100 g) [dry weight sample] among the methanol crude extracts. In phase two, thirty-five healthy male adult rabbits (Oryctolagus cuniculus; New Zealand White strain) with a body weight of 2300–2800 g were used. The rabbits were randomly assigned to one of the seven rabbit groups. Four groups were fed cholesterol diet (1% cholesterol) and 0, 0.5, 1.0, and 1.5 mL of juice per kg of rabbit daily (hypercholesterolemic groups), while the other three groups were fed commercial rabbit pellet and 0, 0.5, and 1.0 mL of juice per kg of rabbit per day (normocholesterolemic groups) for 90 days. Blood samples were taken before and after the experimental period. The serum, aorta and liver homogenates were also analyzed for biochemical biomarkers. The results showed that the physiological dysfunctions of the hemopoietic system, caused by the high-cholesterol diet, were significantly (p<0.01) normalized by the administration of BA whole fruit juice, especially at the highest dose (1.5 mL/kg/day). In phase three, histopathological studies were carried out to evaluate the percentage of atherosclerotic lesion accrued using different staining techniques (Hematoxylin & eosin, van Gieson and Sudan IV). The result showed that the supplementation of high-cholesterol diet of the hypercholesterolemic rabbits with only 0.5 mL BA per kg rabbit per day significantly (p<0.001) attenuated aortic fatty streak development. Higher BA fruit juice doses used (1.0 and 1.5 mL per kg rabbit per day) also significantly decreased further the development of aortic fatty streaks. Therefore, the phytochemical composition and antioxidant properties of BA whole fruit juice have substantially proved its potential health benefits as an effective hypocholesterolemic, anti-inflammatory and anti-atherosclerotic agent for the management of CVD biomarkers. Further studies may be needed to corroborate these facts.

The consumption of fruits has been correlated with better health and prevention of several diseases. Phytochemicals such as phenolic compounds and vitamins in fruits are believed to act as antioxidants that could reduce oxidative damage to biomolecules by modulating reactive free radicals. Since Malaysia is rich in a wide variety of tropical fruits, we should examine them in producing healthy and tasty juices. A new fruit juice named Mixed Fruit Juice (MFJ) was formulated from a combination of three tropical fruits (soursop, mango and kasturi lime).There is no information regarding the antioxidant activity, antioxidant compounds, safety parameters and bioavailability of the bioactive compounds in this product. Therefore, this study focuses on the measurement of antioxidant activity by using the common chemical methods (TPC, FRAP, DPPH) followed by the identification and quantification of phenolic acids by GC-MS. The safety and in vivo antioxidant effects of MFJ were done in normal and aged rats. Human intervention study was established in order to determine the bioavailability of the phenolic compounds in MFJ. Results show that MFJ contained 23.50±1.57 mg GAE/100g (TPC) and 18.77±0.46 mg TE/100g (FRAP) and DPPH value at 83.66±0.84%. The 28-day sub-chronic toxicological test showed no systemic toxicity attributable to the MFJ administration in normal rats. There was no significant effect in the haematological profiles and both liver and kidney function tests when compared to control rats. Furthermore, MFJ was able to modulate the antioxidant enzymes such as SOD, GPx and CAT in the blood and tissues (liver and brain) of aged rats. Aged condition reduces all the antioxidant enzymes in blood and tissues. The supplementation of MFJ in aged rats had increased the TAS, GPx and SOD values in the blood by 14%, 30% and 104%, respectively. In the liver, the CAT and SOD also increased by 33% and 22%, respectively. In addition, substantial increment was observed in the activity of enzymes in the brains of aged rats. GPx was found to increase by 75% in aged rats after receiving MFJ. Moreover, SOD had increased more than 100% in aged rats after receiving MFJ. In the bioavailability study, phenolic acids (p-coumaric acid, ferulic acid and caffeic acid) were found in conjugated form (plasma and urine) and mainly further metabolized to hippuric acid that was excreted in the urine after one hour of MFJ ingestion. Hippuric acid, the main metabolite in urine was found to increase by 25% compared to baseline. In addition, ferulic acid which is known as an antioxidant compound was also recovered in the urine at 16% in one hour. Thus, it is suggested that the absorption of phenolic acids may occur in the small intestine. In conclusion, this current study shows that MFJ was able to exert the antioxidant effects due to the content of phenolic acids which was proven bioavailable in humans.

Food insecurity occurs when there is uncertainty of the availability of food or the insufficiency in quantity and quality of food required for a healthy life style, or the need to use socially unacceptable ways to acquire foods. Based on several studies conducted from 2001 to 2014 in Malaysia, 58% to 86% of households were reported as having some kind of food insecurity. However, none of those studies has mentioned the prevalence of food insecurity in the university student population. Therefore, this study had sought to assess the validity and reliability of a cross-culturally adapted English version of the U.S. Adults Food Security Survey Module (AFSSM) in the measurement of food insecurity among university students. A total of n=208 Malay university students aged between 18-25 years old were selected in this cross-sectional study using convenient sampling. The students’ socio-economic status were evaluated using a self-reported questionnaire. The eleven item AFSSM was used to measure food insecurity. Content validity was assessed by an expert panel, and the questionnaire was then tested among 22 students for clarity. Internal consistency was evaluated by Cronbach’s ? and construct validity was assessed through exploratory factor analysis. Criterion validity was assessed by correlating the measure with a number of selected nutrition and inflammatory biomarkers. In all, the prevalence of food insecurity was measured at 56.3% (95% CI: 49.2%-63.1%), with 21.6% (n=45) of students having very low food security. Cronbach’s ? was observed at 0.81. Factor analysis of the AFSSM items revealed two factors: factor 1 (Insufficient food quantity) and factor 2 (Psychological). For criterion validity, only three items were significantly correlated with measured biomarkers. Item Q2 (r=0.15; p=0.044) and Q4 (r=0.18; p=0.018) were positively correlated with serum hs-CRP. On the other hand, item Q5 (-0.15; p=0.046) was found to be negatively correlated with serum folate levels. Only gender was significantly associated with food insecurity where female students were less likely to be food insecure (OR=0.32; 95% CI: 0.11-0.90; p=0.030). As for the conclusion, the cross-culturally adapted and validated English version of the AFSSM showed acceptable levels of reliability, construct validity, and criterion validity to recommend its use in assessing individual food insecurity among Malay university students.

The difficulty associated with the current techniques drove the investigations presented in the present thesis which aimed at investigating the clinical usefulness of corneal topography and slit lamp videography in the assessment of tear film dynamics in dry eye and contact lens wear.Six-hundred and forty-two (642) eyes of 321 subjects took part in Study 1 where several parameters were derived from corneal topography and slit lamp videography, and the assessment of meibomian gland dysfunction (MGD) and Schirmer test with anaesthesia (STA) made. Tear film break-up time (TFBUT) was significantly correlated with STA (tear production test), MGD (tear production and tear elimination tests), and lipid floating time (LFT) (p < 0.05). Corneal irregularity measurement (CIM) after 5 seconds of holding the blink (CIM 5) had significant correlation with LFT while CIM after 8 seconds of blink holding (CIM 8) was significantly correlated with MGD. Furthermore TFBUT, LFT, and tear meniscus height (TMH) were significantly reduced in the dry eye group compared to non-dry eye group. In Study 2 parameters from slit lamp videography (as derived in Study 1), plus pre-lens tear film (PLTF) non-invasive tear break-up time (obtained from corneal topography) were followed-up in the investigation of the effects of 8 hours of contact lens wear comparing that of each subject’s own habitual hydrogel contact lens to newly-prescribed daily silicone hydrogel contact lens. Thirty six (36) eyes of 18 habitually wearing hydrogel contact lenses (mean age =22.9 ± 3.7 years old) were involved, and it was found that PLTF, TFBUT, and LFT were all significantly reduced (p < 0.05) after wearing each lens type for 8 hours. The tear film changes noted for each lens type after the 8 hours of wear were however, not significantly different (p > 0.05) between habitual hydrogel and the silicone hydrogel contact lenses. In Study 3 the tear ferning patterns (indirect tear production test) were compared among non-contact lens wearing, soft contact lens wearing, and post-contact lens related microbial keratitis (Post-CLRMK) subjects. Abnormalities in the tear ferning pattern of the habitual contact lens wearing subjects and post-CLRMK subjects were statistically similar (p > 0.05). In conclusion corneal topography and slit lamp videography have been shown from the studies presented in this thesis to be useful clinical tests representing the phases of tear production, distribution and elimination, for the assessment of tear film dynamics in the clinical setting.

Severe overfishing and stock depletion of worldwide sea cucumber has encouraged the development of sea cucumber aquaculture. Five species of local microalgae were cultured prior to larval and juvenile rearing as the main food source. Results for mass culture showed that they can be cultured outdoor which yielded a higher volume production than indoor culture. Culture in polythene bag showed a significant difference in growth compared to carboy culture and subsequently resulted in shorter harvest period. Adult H. scabra were collected from wild and subjected to two spawning induction method namely the thermal stimulation and dry-algae stimulation. Thermal stimulation was proven to be the best method by producing about 1.7 million eggs. Early auricularia larvae were observed 48 h after fertilization and were fed on mix microalgal culture. Auriculariae larvae transformed into dolilaria larvae after 11th day fertilization and the non feeding doliolaria metamorphosed into creeping pentactulae after 17th day. The survival rate of 4.2 % was achieved. Juveniles were fed with mixture of microalgae, sea weed and spirulina according to their growth stages. Average growth rate was 0.8 mm per day. Survival rate in the juvenile rearing tank was 32%. After 60 days of tank cultivation, juveniles reached 20 mm and were transferred either to hapanet or outgrow pond at 200 animals/m2. Total loss was observed in the hapanet while good growth was observed in the outgrow pond with survival rate of 36%. Juveniles reached average weight of 270 g after 3 months of outgrow pond cultivation. Growth was generally inversely proportional to the stocking density. Higher survival rate and better growth was observed in pond with lesser stocking density.

This thesis represents an effort by the author to overcome the dearth of Islamic discourses in Medical imaging, a sub-specialisation within the bigger field of Medicine. Driven by the concepts of Islamisation of Knowledge (IOK), integration of Islamic input in the professions, as well as his professional obligation as an academician in an Islamic university, the author posits himself to contribute towards the realisation of the above concepts and enriching the Islamic based literature. Efforts were made done to ascertain the state of affairs of the presence of Islamic perspectives in Medical imaging literature. The results showed a major void in Islamic perspectives in the literature. The term “perspective” and “perspectives” used in this thesis are seen within the definition of perspective; “a particular way of considering something”. The plural form does not suggest that Islam has differing opinions or views when confronted with a particular issue but represents the various sources and types of references. The essence of these individual perspectives remains embedded in Tauhidic (Oneness of Allah) principles and objectives. The thesis proposes a methodology in integrating Islamic perspectives into the theoretical domains within a typical Medical imaging curriculum. Existing knowledge from the predominantly secular environment that has dominated Medical imaging since its inception is not discriminately discarded unless those that is not in line with what Islam champions. Harmonisation and individual tailoring of the Islamic perspectives with the human sciences remain the core approaches in the integration process. The integration also identifies the teaching, learning and assessment considerations. The superficiality in the coverage over the wide dimensions in the discipline is seen within the context of future avenues for in-depth discourses to be made. Typically, the involvement of Muslim practitioners at all levels; the academicians, the practitioners, the leadership, the administration and the professional fraternity are required for promoting, internalising and sustaining Islamic perspectives in the discipline.

Treating articular cartilage defect remains a major orthopaedic predicament. Being a simple structure that is avascular and aneural, articular cartilage has little ability to repair itself when damaged. Cartilage tissue engineering provides an alternative technique for restoring and regenerating damaged cartilage tissues. This study attempts to (1) evaluate and compare the growth of bone marrow mesenchymal stem cells (BMSCs) aspirated from rabbits’ iliac crest and femur in two different culture media; α-Minimum Essential Medium (α-MEM) and a mixture of 1:1 Ham’s F12 Nutrient Mixture (F12) and Dulbecco’s Modified Eagle Medium (DMEM) or FD, (2) to evaluate the in vitro and in vivo cartilaginous tissue formation using BMSCs-seeded poly(lactic-co-glycolic acid) PLGA/Fibrin scaffolds and (3) to explore the bioethical issues within the Islamic perspective in relation to tissue engineering and regenerative medicine (TERM). Growth kinetic assessments were performed on monolayer-cultured BMSCs harvested from two locations and cultured in two different media. Constructs fabricated from BMSCs seeded onto PLGA/Fibrin were cultured for three weeks and thereafter were implanted in rabbit model. The in vitro cartilaginous engineered constructs were evaluated by gross inspection, cell proliferation assay, gene expression, sulphated glycosaminoglycan (sGAG) production and histology at week 1, 2 and 3 while the in vivo construct were harvested at week 6 and week 12 post-implantation. It was observed that BMSCs harvested from the iliac crest region and cultured in FD significantly promoted BMSCs growth. The in vitro study showed that after 3 weeks of culture, the PLGA/Fibrin construct exhibited significantly higher cell viability, higher sGAG content and better histo-architecture and cartilaginous extracellular matrix (ECM) compound in concert with the positive glycosaminoglycan accumulation when compared to the PLGA only construct. Post implantation, the osteochondral defects treated with PLGA/Fibrin/BMSCs constructs showed better repair, more cartilaginous extracellular matrix, higher sGAG content, superior compressive strength and greater expression of chondrogenic marker genes than PLGA/BMSCs group. COL2A1 which is the specific cartilage marker, ACAN, COL9A1 and SOX9 genes were expressed both in the in vitro and in vivo. This study suggested that PLGA/Fibrin when seeded with pluripotent BMSCs that underwent optimum manipulation may serve as a prospective construct to be developed as functional tissue engineered cartilage. This study may also act as a platform to spark the initiative in integrating Islamic bioethics in providing “middle of the road” approach moderating between science and religion. Debates on TERM must comprise on religious and ethical view considering there are still many grey areas that requires Islamic input. Based from the Islamic perspective, scientific researches in TERM are permitted, as long as it is not a threat to human being and applied within the permissible limits described by the Shari’ah.

This thesis aimed to investigate the effect of different pterygium morphology on anterior corneal curvature and predicting visual outcome after surgical intervention. A total of 93 primary pterygium participants were selected from patients who visited an ophthalmology clinic. This prospective cohort study involved pre and post-surgical assessment, with a total of 5 data collection sessions (Pre-surgical, 1, 3, 6, 12 months post-surgical. For pre-surgical, pterygium morphologies (redness, thickness, length, corneo-pterygium total area and dry weight were measured objectively. The association between pterygium morphologies (predictive factors) and predictive outcomes (SimK, CIM, SF, TKM, BCVA and CSF) were investigated. All participants underwent similar pterygium surgery procedures performed by a single surgeon. For all post-surgical assessment, identical measurements of topographic and clinical changes were made based on changes in its predictive outcomes. Comparisons in the magnitude changes between different types of pterygium were performed. Our result shows that fibro-connective components of pterygium were found able to provide the highest prediction with 27 – 55% of the predictive outcomes. With regards to types of pterygium, type III (fleshy) was found causing the biggest changes, followed by type II and I. Concisely, we found that single predictive factor is inadequate to either describe or predict changes on anterior corneal curvature and predicting visual outcome. We had demonstrated that utilizing pterygium morphologies, we were able to describe and predict changes in different types of pterygium better. We suggest that different types of pterygium would give rise to different magnitude changes on anterior corneal curvature and predicting visual outcome, thus we need to consider all pterygium morphologies in assessing pterygium.

Tissue engineering and regenerative medicine (TERM) offers an alternative approach for the treatment of intervertebral disc (IVD) degeneration. One of the important TERM principles is the application of biomaterial scaffolds. Poly(lactic-co-glycolic acid) (PLGA) is a US Food and Drug Administration (FDA) approved synthetic copolymer for clinical use. However, for TERM application, PLGA needs to be improved. Natural polymers, for example atelocollagen and fibrin can be used to enhance the PLGA properties. The combination of PLGA with atelocollagen and fibrin for IVD regeneration has not been explored yet. The aim of the present study is to evaluate firstly the monolayer culture of IVD cells, secondly the PLGA-based scaffolds namely PLGA alone, PLGA with atelocollagen (PA), PLGA with fibrin (PF) and PLGA with the combination of both atelocollagen and fibrin (PAF), thirdly the in vitro 3D constructs engineered from PLGA-based scaffold seeded with the annulus fibrosus (AF) cells and fourthly the in vivo constructs implanted subcutaneously on the athymic nude mice model. Important Islamic principles in relation with the IVD regeneration were also discussed as the fifth objective. Growth kinetics analysis and cellular proliferation assay were performed on the monolayer cells culture and in vitro constructs at day 4, 7, 14 and 21, respectively. Transmission electron microscopy (TEM) and scanning electron microscopy (SEM) were used to evaluate cell morphology both in monolayer and the in vitro 3D culture. The fabricated PLGA- based scaffolds were characterized in terms of chemical bonding, porosity, water uptake, thermal, mechanical, degradation, pH level and compression modulus. Other evaluation parameters included histology, immunohistochemistry, gene expression, sulphated glycosaminoglycan (sGAG) and DNA quantification analysis. All constructs were evaluated at 1, 2 and 3 weeks in vitro and, at 4 weeks in vivo. The presence of autophagasome in AF cells was detected in all passages. Comparable morphology of cellular attachment on in vitro 3D constructs was observed. Passage 1 of AF cells showed better cellular profile and extracellular matrix production compared to all other passages thus chosen to be seeded onto PLGA-based scaffolds and form constructs. The PA scaffolds showed better swelling ratios, thermal stability, mechanical strength, degradation properties and ability to reduce the pH effect. As for in vitro 3D constructs, PAF exhibited higher cellular proliferation and depicted better histological staining than the other groups. Positive accumulation of collagen type I and type II with higher level of sGAG were also observed in PAF constructs. Comparable mRNA expression of condrogenic markers were detected in all monolayer, in vitro 3D constructs and in vivo samples. In line with in vitro 3D constructs, in vivo constructs especially PAF group showed notable distribution of cells, accumulation of sGAG, and proteoglycan with positive immunoreactivity of collagen type I and type II. In conclusion, the incorporation of atelocollagen, fibrin and both combination significantly improved PLGA scaffold properties in vitro and in vivo. The desk-based study on Islamic perspective indicated that TERM technology does not violate any principles of Islam. More works on IVD regeneration are needed to uncover its full potential. This laboratory grown IVD may serve as an alternative over conventional treatments in future clinical application.

Excessive sugar intake has been widely associated with the increased prevalence of obesity and diabetes due to its potential in instigating positive energy balance. Sugar craving, an issue which has yet been explored in our local community, is also postulated to relate to erratic eating behavior, impulsive food intake and binge eating. The initial part of this research aimed to develop and validate a sugar craving assessment tool suitable for Malaysians. Eventually, the Sugar Craving Assessment Tool for Malaysian (MySCAT) was created that has good internal consistency (? = 0.866) and the score was significantly correlated with sugar intake measured via FFQ Sugar (r = 0.332, p <0.01). As opposed to high energy density and palatability of sugar in foods, the low calorie sweeteners (LCS) has been suggested as alternative to sweeten foods but without markedly excessive calories. Stevia has recently been popular as sugar substitute and it is from plant source, hence deemed more acceptable to be consumed by lay person, relative to artificial sweetener like aspartame. Further, we compared the sweetness perception and acceptance of solid and liquid foods containing either sugar, stevia or aspartame. Healthy adults perceived sugar sweeter than both LCS. The replacement of sugar with LCS was acceptable among the subjects. In the subsequent study using different group of subjects, we investigated the association between sugar craving and sweetness perception as well as eating behavior (restraint, emotional and external traits). Results indicate that female, overweight/obese individuals and restraint eaters possessed higher sweetness perception than their counterparts. The final part of this research aimed to compare the effects of sugar, stevia and aspartame consumption on energy compensation, appetite responses (satiety, fullness and desire to eat sweet) and appetite hormones changes (insulin, leptin and ghrelin). The “preload paradigm” method has been used, whereby 3 types of preloads (sugar, stevia and aspartame) were given prior to test meal served as lunch buffet with ad-libitum intake. We found that following LCS preloads, energy intake during test meal was higher (COMPX 45.6% for stevia and 54.1% for aspartame) but total energy intake (preloads + test meal) was significantly higher with LCS consumption. Sugar preload significantly increased leptin the most compared to LCS after 2 hours. There was no significant relationship recorded between sweeteners and their impact on appetitive responses. Conclusively, we had successfully develop a novel method to measure sugar craving among Malaysian and it is useful in the assessment of the risk of excessive sugar intake. In the effort of reducing energy density of food and sugar content, replacement of sugar with stevia, a natural sweetener, is recommended as it proven to reduce total energy intake while maintaining the normal hormonal changes and do not interfere with appetitive responses.

Scattered gamma photons are the result of Compton scattering interaction of radiation within the matter. They have low energy as compared to unscattered photons and their contribution is adverse in terms of degradation of spatial resolution and image quality in single photon emission imaging. The objective to apply a physical filter is to reduce the amount of scattered gamma photons before reaching the scintillation detection system. The physical filter material aluminium, Al sheet (0.10mm and 0.20mm thickness) was chosen to perform this study. The type and thickness of physical filter was selected on the basis of percentage attenuation calculations of different gamma ray energies by various thicknesses and materials. The parameters that were investigated in this study are spectra of Tc-99m, spatial resolution, uniformity and system volume sensitivity. For the specific organ study, the image acquisitions from the Hoffman 3-D brain phantom and the RSD Striatal brain phantom were investigated. Data were acquired using Infinia® GE Healthcare dual-head gamma camera without and with physical filter with LEHR collimator installed. Spatial resolution study was done by scanning a Tc-99m line source (0.8mm inner diameter) at various source-to-collimator distances in air and in scattering medium. For spectra, uniformity and sensitivity, a cylindrical source tank filled with water added with Tc-99m was scanned. Both Hoffman 3-D and RSD Striatal brain phantoms with water with added Tc-99m were scanned. The Hoffman 3-D brain phantom was prepared with two conditions; normal brain and cold defect brain. Both brain phantoms images were analysed qualitative (visual analysis) and quantitatively (grey-to-white ratio, thalamus size and contrast ratio). All SPECT images were reconstructed with filtered-back projection (FBP) method by applying Butterworth filter of order 5 with different cut-off frequencies 0.30, 0.40 and 0.50 cycles/cm. The images were corrected by Chang’s attenuation correction method using linear attenuation coefficient, LAC 0.11cm-1 and 0.12cm-1 for without physical filter and 0.12cm-1, 0.13cm-1 and 0.14cm-1 for with physical filter. The spatial resolution with the physical filter improved as the source-to-collimator distance increases. There was a significant reduction in count rate from Compton and photopeak regions of Tc-99m spectra with physical filter compared to without physical filter. There was a substantial reduction in the counts from the photopeak region where the decrease in the scatter radiation is probably higher. System volume sensitivity was reduced. Despite reduction in system volume sensitivity, the system uniformity improved with physical filter. Enhancement in image quality of Hoffman 3-D brain phantom and the RSD Striatal brain phantom by applying 0.10 mm Al physical filter was achieved. Therefore, this technique is considered suitable for further investigation for clinical trials to validate the applicability of physical filter technique in clinical SPECT imaging.

Myocardial SPECT is one of the techniques that provide high diagnostic accuracy for the assessment of coronary artery disease. However, in general, the presence of Compton scattered photons in the resulting image data will reduce the diagnostic accuracy. This undesirable effect is even more significant in heart thorax-linked analysis compared to other regions with a more homogenous volume. In this study, the use of physical filter was proposed to reduce the undesirable scattered gamma photons. In this context, the scattered gamma photons were absorbed by the filter before they reach the gamma camera detector. Initially, a selection of the types and thickness of the filter materials based on a theoretical calculation was done. They were then incorporated into the basic procedures of Planar and SPECT imaging, and finally to Myocardial SPECT imaging, where a Tc-99m radionuclide was used. In Myocardial SPECT imaging procedure, an anthropomorphic torso (heart/thorax) phantom was used. The image reconstruction procedure was based on filtered back projection. Chang’s attenuation correction method was used. The image quality was analyzed qualitatively and quantitatively. In Tc-99m spectra test, a decrease in the ratio of scattered to non-scattered photon for the whole spectra was observed for all types of the physical filters examined. However, the reduction in photopeak region was only recorded by Zn 0.2 mm filters. The physical filters were also found to improve system’s spatial resolution, but the uniformity of the tomographic image was unchanged, and the system volume sensitivity was reduced to 16% for Cu and 4% for Zn filter. It was found that Zn 0.2 mm reduced the scatter in Myocardial SPECT imaging, where a clear separation between the liver and heart was observed. A significant improvement in contrast (10.98%) and signal-to-noise ratio of myocardial wall to defect areas (12.68%) was achieved with the use of Zn 0.2 mm material filter. Thus, it is concluded that the use of Zn 0.2 mm material filter has the potential to enhance the image quality in clinical SPECT imaging. However, clinical trials of this technique are required prior to its use in patient studies.

Azoospermia is a male infertility worldwide concern due to incomplete spermatogenesis process. Recreating spermatogenesis outside of its original environment (in vitro) is a scientific curiosity in andrology world. However, it remains challenging due to the limitation of culture system. Testicular biopsy cells from non-obstructive azoospermia (NOA) (complete absence of spermatozoa) and obstructive azoospermia (OA) (obstruction in the male ducts results in absence of spermatozoa in semen) patients were obtained to develop in vitro spermatogenesis. Modified human embryonic stem cells (HESC) media using knockout DMEM and knockout serum replacement were used to determine growth factors (basic fibroblast growth factor (BFGF) and leukemia inhibitory factor (LIF)) that were suitable for the development of spermatogenic cells. In the early phase of study, NOA sample was selected to see the potential development of spermatogonial stem cells (SSCs). The sample was cultured in HESC medium with BFGF. Protein markers; ITGA6, ITGB1, GFRA6 and CD9 were done using immunofluorescent staining on Day 1, 7, 14 and 21 but non of the markers were present, only unknown cells has been detected. Cultures were then extended until Day 49 using both NOA and OA samples. Each sample divided into two groups; HESC with BFGF and HESC with LIF. OCT4, ITGA6, ITGB1, GFRA6 and CD9 markers were positive in immunofluorescent staining and quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) indicated the SSC-like cells development in both NOA and OA samples. Both of the culture samples were extended until 90 days and most of the azoospermia samples had successfully developed post-meiotic cell specified spermatid-like cells except NOA sample cultured with HESC and BFGF that shown unknown cells detected. This revealed that late spermatogenesis could be established in vitro using HESC or in vitro fertilization (IVF) media with the addition of reproductive hormones (follicle stimulating hormone (FSH) and testosterone). SCP3, H2B and TP1 markers were positive indicating that meiotic division has occurred in the culture. This study managed to show some evidence of in vitro spermatogenesis. It opens of possibilities to create spermatozoa in the future, thus giving hope to azoospermic especially NOA patients to have biological children.

Cancer is a major cause of morbidity and mortality worldwide. In recent biomedical researches, the areas of cancer and infectious diseases have a leading position in utilization of medicinal plants as a source of drug discovery. Malaysia has a diversity and large quantity of underutilized fruits which are rich in phenolic compounds. The objective of study one was projected in vitro to explore natural sources of antioxidant in Artocarpus.altilis (breadfruit) extracts and antioxidant properties. The total phenolic content (TPC) was measured using the Folin-Ciocalteau method and the total flavonoid content (TFC) was determined by using aluminium chloride colorimetric method. Antioxidant properties were determined via the DPPH radical scavenging and β–carotene bleaching (βCB) assays. The various fruits parts Pulp (PU), peel (PE) and whole fruit (WF) were extracted with various solvents such as hexane, dichloromethane (DCM) and methanol. The methanol extracts obtained the highest yields among other solvents (hexane and DCM). The pulp (edible portion) had the highest yield (p<0.01). Methanol extract of pulp part revealed the highest total phenol and flavonoid content value of 781±17.32 mg (GAE)/g and 6213.33±82.24 mg (QE)/g of dry sample, respectively. IC50 values of methanol extract of pulp part in DPPH radical were obtained to be 0.05±0.00 mg/mL as compared to positive control (ascorbic acid) 0.06±0.00 and the antioxidant activity for the β-carotene bleaching assay was 88.34±0.75% of methanol extract of pulp part as compared to the positive control (Trolox) 90.02±0.87%. The objective of study two was to identify and quantify some phenolic compounds in the methanol extracts. By using the ultra high-performance liquid chromatography-tandem mass spectrometry (UHPLC/MSMS) based approach, a total of 9 compounds were detected and characterized on the basis of their chromatographic retention time, UV-vis spectra and mass spectra in the negative-ion mode and data from the literature. The results of the various parts of A. altilis fruit extracts showed promising antioxidant and potential bioactivities due to the high content of phenolic compounds. The purpose of the study three was to evaluate the cytotoxicity effects of methanol fruit extracts on four human cancer (A375, MCF-7, A549, and HT-29) cell lines. The IC50 of the samples were measured using trypan blue exclusion assay (TBEA). The methanol extract of pulp part showed the least inhibition concentration of 15.40±0.91 μg/mL on A375 cells. In the study four, the molecular mechanism of methanol extracts-induced apoptosis and cell cycle arrested in human cancer cells were investigated in a time dependent approach by using flow cytometry. The treated cells were stained with nexin to detect early and late apoptosis and with PI for cell cycle arrest associated with the DNA fragmentation, various cells arrests were occurred at G1/S, S and G2/M phases. Lastly the gene expression analysis by reverse transcription quantitative PCR (qPCR) method was carried out by analysing the expression of gene of interest for quantification of mRNA levels. Results after cells treated with IC50 were revealed by upregulating of anti-apoptotic genes/downregulated of pro-apoptotic BCL-2 gene expressions were triggered the treated cells into CASPASE-3, intrinsic and extrinsic pathways. These findings suggest that the methanol extracts of three parts of A. altilis fruit have potential chemotherapeutic activity against human cancer cell lines mainly the pulp part of the fruit.

The invasiveness of malignant melanoma is mainly attributed to the enzymatic destruction of the extracellular matrix and basement membrane components by a group of enzymes known as matrix metalloproteinases (MMPs). The expression of these proteinases is mainly regulated at the transcriptional level; therefore, high expression of MMPs is manly attributed to different transcription factors which enhance or inhibit the promoter activity of MMPs genes. Among these factors, YB-1 and CTCF proteins are transcription factors in which CTCF is mainly a tumour suppressor protein; while YB-1 is an oncogenic factor and a prognostic indicator in a wide range of tumours that regulates most of the cancer processes such as proliferation, invasion and metastasis by regulating the expression of genes related to those processes. However; the expression of these transcription factors and their potential effect on the expression of collagenases MMPs in malignant melanoma cells are not yet confirmed. Therefore, this study was conducted to determine the expression of collagenases MMPs (MMP1, MMP8 and MMP13), YB-1 and CTCF transcription factors in A375 melanoma cancer cells. In addition, the stromal effect of normal skin fibroblasts on the expression of collagenases and proliferation in A375 cell was determined. The results of this experiment demonstrated an increase in the expression of YB-1, MMP8 and MMP13 in A375 cells. Thereafter, this was followed by the establishment of YB-1 silenced strain of A375 cells using a silencing short hairpin RNA (shRNA) construct. The effect of YB-1 knockdown on the expression of collagenases MMPs was determined using reverse transcription PCR and Western blotting. In addition, the antiproliferative effect was determined using flow-cytometry, colorimetric MTT assay and cell counting; while the anti-invasive properties were determined using wound healing assay. The results of this experiment elucidated that YB-1 protein regulates the expression of MMP13, cell cycle progression, cell proliferation and cell migration of A375 cancer cells in vitro. Therefore, the direct interaction between YB-1 protein and the AP-1 promoter sequence of MMP13 was evaluated using Chromatin Immunoprecipitation assay (ChIP). The ChIP analysis has confirmed no interaction between YB-1 protein and the AP-1 promoter sequence. Finally, these experiments demonstrated YB-1, MMP8 and MMP13 were highly expressed in the A375 cancer cells. The stromal cells were found to promote A375 cell proliferation and enhance the expression of MMP1. In addition, YB-1 silencing was significantly associated with reduced expression of MMP13 enzyme, reduction in cancer cell proliferation in a cell cycle specific manner and anti-invasive properties. Therefore, YB-1, MMP13 and stromal cells are considered as promising elements that might help as a potential target in the treatment of melanoma tumour due to their roles in the processes of invasion, migration and proliferation. Further experiments are needed to demonstrate novel protein partners of YB-1 and novel binding sites within gene promoter region of MMP13 with determination of the involved signalling pathways.

Given the beneficial roles of adiponectin protein on body metabolism and its profound protective effects against metabolic diseases, a better understanding of the protein’s secretion and regulation is very important. The objectives of this study were to extract, detect and quantify the total adiponectin in trimmed off abdominal adipose tissues from meat sources, namely chicken, beef and lamb, and test it on STZ-induced hyperglycaemic rats. Abdominal adipose tissues were isolated from the aforementioned sources and delipidation of the tissues were performed through chloroform/methanol extractions. Afterwards, the protein concentration was determined by using Protein Assay Bicinchoninate Kit method. This was followed by quantification of the adiponectin using ELISA assay kit. Followed by tests in STZ-induced hyperglycaemic rats on blood glucose and blood lipid, hormone measurement and lastly quantification of PPAR-? mRNA, AMPK mRNA, AdipoR1 mRNA, AdipoR2 mRNA and AdipoQ mRNA in the hepatocytes. The experiment was conducted in triplicates and the results were presented as mean ± SD. The data was statistically analyzed by using SPSS statistical software version IBM 21.0. One-way analysis of variance (ANOVA) was used and the data were considered statistically different at 95% confidence interval. Results indicated that the extraction of 10-gram subcutaneous adipose tissues from chicken, beef and lamb yielded 0.10-gram, 0.15 gram and 0.15 gram of protein, respectively, which were 1 - 1.5 % from the total tissue mass. The protein concentration in the abdominal adipose tissues from chicken, beef and lamb were 1.25 ± 0.05, 1.75 ± 0.05 & 2.53 ± 0.07 mg/ml, respectively. The isolated adiponectin concentration in chicken, beef and lamb was 158 ± 0.05 ng/ml, 24240 ± 0.05 ng/ml and 37 ± 0.08 ng/ml, respectively. Adiponectin concentration in beef abdominal adipose tissues was significantly (p<0.001) higher compared to chicken and lamb. In the animal study, the normal, insulin-treated, PCCA-treated (protein containing chicken adiponectin), PCBA-treated (protein containing beef adiponectin) and PCLA-treated (protein containing lamb adiponectin) hyperglycaemic groups exhibited significantly (p<0.05) reduced blood glucose levels after the treatment period when compared with the NT (control diabetic) group. Similarly, the normal, insulin-treated, PCBA-treated and PCLA-treated diabetic groups exhibited significantly (p<0.05) reduced in blood cholesterol and triglycerides levels after the treatment period when compared to the NT group. Moreover, serum adiponectin concentration was significantly (p<0.05) higher in the normal, insulin-treated, PCBA-treated, PCCA-treated and PCLA-treated diabetic groups as compared to the NT group. Nevertheless, serum insulin concentration was significantly (p<0.05) higher in the normal, insulin-treated and PCBA-treated groups as compared to the NT group. Lastly, AMPK, AdipoR1 and AdipoR2 mRNAs were significantly (p<0.05) upregulated in the normal, insulin-treated, PCBA-treated, PCCA-treated and PCLA-treated diabetic groups as compared to the NT group. However, there was no significant difference among groups for PPAR-? mRNA and AdipoQ mRNA. In conclusion, the present study suggested that the beef protein had the highest amount of adiponectin protein and gave the highest positive effects on STZ-induced hyperglycaemic rats. Thus, adiponectin proteins extracted from these cheaper sources of wasted adipose tissues in meat can be one of the promising target for future novel pharmacological and therapeutic treatments/preventions for insulin resistance and metabolic diseases.

Malnutrition is one of the most distressing public health problems all over the world. Undernutrition is common in third world countries. It is an insufficiency of nutrients, minerals, and vitamins, that person requires to keep up the best possible strength of cells, tissues and organs. In the previous two and half decades (1990 – 2015), a great effort toward accomplishing millennium development goal (MDG) four were performed. The number of under-five mortality significantly decreased by half during this period. However, a significant number of children less than five years are still suffering and dying because of undernutrition in developing world. This lead to a negatively influences on physical wellbeing and linear growth of infants and pre-school children. Ethiopia is one of the poorest among the developing nations and undernutrition is the highest within the sub-Saharan Africa. Hence, this study aims to evaluate the effect of nutrition education intervention (NEI) on knowledge, attitude and practice (KAP) of the mothers/caregivers on infant and young child feeding (IYCF) behaviours and nutritional status of the children below five years. This study was conducted in two phases, first phase was a cross sectional study assessing mothers/caregivers KAP on IYCF and nutritional status of the children, in order to identify the gaps, plan properly, and conduct NEI. In the second phase a group randomized controlled trial design were employed. When selecting the intervention and control districts the socio-demographic, economic and life style similarities were taken into consideration. A total of 415 in phase one and 404 in phase two child-mothers/caregivers pairs participated in the study from Gode and Adadle districts of Shabelle zone. Semi structured interviewer administered questionnaire, anthropometric, blood sample assessment were performed. At baseline study using multivariable logistic regression showed being in Gode District (p <0.001), and not being housewife (p <0.05) were significantly associated with having good knowledge about optimal infant and young child feeding (IYCF), and wealth index (p <0.01) were positively associated with favourable attitude, and being literate (p <0.01) is positively associated with good practice. At post intervention, the KAP of mothers/caregivers mean score improved significantly after NEI. Paired sample t-test showed that KAP mean score of mothers/caregivers significantly (p <0.001) improved after intervention. Similarly, independent t-test mean score difference of the difference (DOD) were significantly improved (p <0.001) in all three variables compared to control group. Likewise, the mean Z-score of wasting (p <0.001) and underweight (p <0.01) of the children within the group were significantly improved in intervention group. This was not seen in stunting, which was almost similar in both phases. Nonetheless, control group showed a significant worsening mean Z-score for wasting, underweight, and stunting (p <0.05) that indicates the worsening of the children nutritional status. Regarding the anaemia status of the children, the mean score showed statistically significant improvement (p <0.001), after NEI in both groups. But, the intervention group has significantly higher increment. In conclusion, NEI had shown improvement of maternal/caregivers KAP related to IYCF behaviours. Thus, subsequently improved the nutritional status of the children. Therefore, NEI should be scaled up and expanded to rural communities in sustainable manner, an especial attention should be given to those families and communities with poor wealth index and food insecure by provision of accessible, available and affordable food items and services at grass root level.

Aquilaria malaccensis (AM) or locally known as ‘gaharu’ (agarwood) is a species of Aquilaria genus and belongs to the Thymelaeaceae family. It is widely distributed in Malaysia, Indonesia, and the Borneo Islands. Traditionally, its leaves were used to relieve bruises and studies have shown that they function as an antioxidant, aphrodisiac, and tranquilizer. Despite its proven beneficial medicinal properties, information regarding its toxicity is limited. Therefore, this study was conducted to investigate the male reproductive toxicity of AM. Prior to the toxicity study, the quality and antioxidant property of standardized A. malaccensis leaves aqueous extract (AMLAE) was determined by a set of phytochemical screening, High-Performance Liquid Chromatography (HPLC) analysis, heavy metal, microbial contamination and in vitro antioxidant tests. The general toxicity of AMLAE was evaluated based on acute and sub-acute oral administration in Sprague Dawley (SD) rats according to the Organization for Economic Cooperation and Development (OECD) Guideline 420 and 407 respectively. The OECD Guideline 421 was selected to evaluate the male reproductive toxicity analysis which comprised of control group and three AMLAE-treated groups (100, 300 and 500mg/kg) respectively. In total of 63 days of oral administration was carried out prior to the one-to-one mating activity with female. Male necropsy was conducted upon sperm-positive vaginal smear to evaluate the male reproductive parameters. Pregnant female rats were necropsied on day 21 to evaluate the reproductive outcomes via caesarean hysterectomy. The extraction protocol successfully yielded 17.64% powder extract. Phytochemical analyses revealed the presence of saponins, phenolics, tannins, flavonoids and aromatic compounds. No microbial and heavy metal contamination was detected. HPLC analysis of the AMLAE revealed that it contained mangiferin (31.08mg/g) as one of its major constituents. AMLAE indicated strong cupric ion reducing power and potent scavenging activity with 740.83mmol Trolox equivalent/g and 1.24 ± 0.27μg/ml respectively. The assessment of acute toxicity revealed that AMLAE did not influence mortality, clinical behaviours, body weight gain, or necropsy findings at a dose of 2000mg/kg body weight. In the sub-acute toxicity, both male and female rats had shown abnormalities in the liver and kidney histology at the dose of 2000mg/kg. No significant findings were recorded in male reproductive parameters and reproductive outcomes on pregnant rats except significant elevations in the in vivo antioxidant activity, hormonal concentration, testicular histology, protamination level and protamine 1 gene expression. Data from present results revealed that AMLAE did not exhibit toxicity on male reproductive system and the no observed adverse effect level for male reproductive toxicity was ˃500mg/kg via oral route.