Introduction: Microbial keratitis is one of the most challenging complications of contact lens (CL) wear. Proper CL practice plays an important role to reduce the risk for contact lens related microbial keratitis (CLRMK). Methods: This multi-centre case-control study was conducted from January 2008 until June 2009 to determine the risk factors associated with CLRMK. Cases were defined as respondents who were treated for CLRMK, whilst controls were respondents who were contact lens wearers without microbial keratitis. Ninety four cases were compared to 94 controls to determine the risk factors for CLRMK. Results: The predictors for CLRMK were: Not washing hands with soap before handling CL (aOR 2.979, CI 1.020, 8.701 p=0.046), not performing rubbing technique whilst cleaning the CL (aOR 3.006, CI 1.198, 7.538 p=0.019) and, not cleaning the lens case with multipurpose solution daily (aOR 3.242 CI 1.463, 7.186 p=0.004). Sleeping overnight with the CL in the eye (aOR 2.864, CI 0.978, 8.386 p=0.049) and overall non-compliance with lens care procedures (aOR 2.590, CI 1.003, 6.689 p=0.049) contributed significantly to CLRMK. Conclusion: Health education and promotion in contact lens care are important and should be conducted by eye care practitioners to reduce the occurrence of CLRMK.
Contact Lenses

Introduction: Current prognostic markers have improved survival prediction, however, it has not advanced treatment strategies. Gene expression profiling may identify biological markers suitable as therapeutic targets. Leukaemia stem cell is associated with adverse outcome, however, its biological characteristics are still being investigated. We observed higher in vitro cell viability in acute myeloid leukaemia (AML) samples with poor prognosis, which may be stem cell related. Objective: The objective of this study was to profile highly expressed genes in an AML sample of poor prognosis/high viability and compare with a sample of good prognosis/low viability. Method: Subtractive hybridization was performed on two AML samples with high blast counts (>80%), a poor prognosis, PP (disease free survival, DFS<12 months) and a good prognosis, GP (DFS>12 months) sample. The PP sample had higher CD34+ counts (73% vs 46%) and higher cell viability than the GP sample. cDNA libraries were subsequently cloned and sequenced. Results: cDNA subtracted from the PP samples was identified as genes active during fetal/embryonic development (LCOR, CNOT1, ORMDL1), HOX- related genes (HOXA3, PBX3, SF3B1), hematopoiesis (SELL, IL-3RA) and aerobic glycolysis/hypoxia (PGK1, HIGD1A) -associated genes. Majority of GP clones isolated contained genes involved in oxidative phosphorylation, OXPHOS (COXs, ATPs, MTND4 and MTRNR2), protein synthesis (including ribosomal proteins, initiating and elongation factors), chromatin remodeling (H2AFZ, PTMA), cell motility (MALAT1, CALM2, TMSB4X), and mitochondria (HSPA9, MPO) genes. Conclusion: Thus, the PP sample exhibited stem cell-like features while the GP sample showed cells at a high level of cell activity. These genes are potential prognostic markers and targets for therapy.
Leukemia, Myeloid, Acute

Defatted dabai peel contains a high amount of anthocyanin. Anthocyanins are known to prevent several types of disease, including cardiovascular-related complications. This study aimed to describe the effects of different doses of defatted dabai peel extract by histopathological analyses on lesions in the liver, kidney, heart and aorta. Histopathology methods were applied to determine the protective effects of defatted dabai peel extracts against hypercholesterolemia-induced oxidative damages to animal organs. Haematoxylin and eosin staining was applied for histopathology examination for liver, kidney, heart and aorta. Data showed that a high dose of defatted dabai extract (3000 mg per day) applied to hypercholesterolemic rabbits for eight weeks had mild protective effect, especially reducing the severity of hepatic fibrosis and steatosis of the renal medulla. The high dose of extract supplementation also reduced inflammation of aorta and formation of atherosclerosis plaque in the cell wall of right ventricle of the heart. The high dose of defatted dabai peel extract could be a protective agent against oxidative stress.
Anthocyanins

Azadirachta indica (neem) has been used for a long time in agricultural and alternative medicine. Neem had been proved effective against certain fungi that could infect human body. This pilot study aims to demonstrate the antifungal effect of Malaysian neem leaf extracts on the pathogenic fungi in otomycosis, Aspergillus niger and Candida albicans. This is a laboratory-controlled prospective study conducted at Universiti Sains Malaysia. The powder form of Malaysian neem leaf was prepared. Ethanol and aqueous extracts of the neem leaf was diluted with sterile water to establish five different concentrations of 50 g/ ml, 25 g/ml, 12.5 g/ml, 6.25 g/ml and 3.125g/ml. The extract was tested on Sabouraud Dextrose Agar suspended with Candida albicans and Aspergillus niger respectively. Well diffusion method was used and zone of inhibition was measured. Growth of the fungi was inhibited in both alcohol and aqueous extract concentrations. The minimum inhibitory concentration (MIC) of Malaysian neem aqueous extract against Candida albicans was 11.91 g/ml, neem ethanol extract against Candida albicans was 5.16 g/ ml, neem aqueous extract against Aspergillus niger was 7.73 g/ml and neem ethanol extract against Aspergillus niger was 9.25 g/ml. Statistical analysis showed that the antifungal activity of Candida albicans is better in alcohol neem than aqueous extract (p<0.001) but aqueous neem extract is better than alcohol extract (p<0.001) for Aspergillus niger.Malaysian neem has significant antifungal effect towards Aspergillus niger, best in aqueous extract and towards Candida albicans, was best in alcohol extract.
Azadirachta

With advancement in genetic studies, familial phaeochromocytoma (PCC) and paraganglioma (PGL) are increasingly being recognized. Characteristically, correlations exist between genotypes and clinical and biochemical phenotypes. We report a phaeochromocytoma in a young patient with intriguing family histories, raising the possibility of his being a familial case.
Genetic Association Studies

Extracranial aneurysms are a rare presentation accounting for only less than 5% of all peripheral artery aneurysms. The left common carotid artery aneurysm is considered even rarer. We present a case of a 46-year-old gentleman who presented to casualty unit with a neck mass, stridor and acute respiratory distress. Prior to this admission patient was being investigated for Takayasu arteritis (TA). Patient was subsequently intubated due to respiratory compromise. A CT angiography of the thorax was done showed a proximal left common carotid artery aneurysm with contained leakage of aneurysm with severe compression of the trachea. Patient was subsequently transferred to the vascular team in National Heart Institute for further management.
Aneurysm

Introduction: The phenotype and genotype of cancer cells portray hallmarks of cancer which may have clinical value. Cancer cell lines are ideal models to study and confirm these characteristics. We previously established two subtracted cDNA libraries with differentially expressed genes from an acute myeloid leukaemia patient with poor prognosis (PP) and good prognosis (GP). Objective: To compare gene expression of the leukaemia associated genes with selected biological characteristics in leukaemia cell lines and normal controls. Methodology: Expression of 28 PP genes associated with early fetal/embryonic development, HOX-related genes, hematopoiesis and aerobic glycolysis/ hypoxia genes and 36 GP genes involved in oxidative phosphorylation, protein synthesis, chromatin remodelling and cell motility were examined in B-lymphoid (BV173, Reh and RS4;11) and myeloid (HL-60, K562) leukaemia cell lines after 72h in culture as well as peripheral blood mononuclear cells from healthy controls (N=5) using semi-quantitative polymerase chain reaction (PCR) method. Cell cycle profiles were analysed on flow cytometry while MTT cytotoxicity assay was used to determine drug resistance to epirubicin. Results: Genes expressed significantly higher in B-lymphoid leukaemia cell lines compared to healthy controls were mostly of the GP library i.e. oxidative phosphorylation (3/10), protein synthesis (4/11), chromatin remodelling (3/3) and actin cytoskeleton genes (1/5). Only two genes with significant difference were from the PP library. Cancer associated genes, HSPA9 and PSPH (GP library) and BCAP31 (PP library) were significantly higher in the B-lymphoid leukemia cell lines. No significant difference was observed between myeloid cell lines and healthy controls. This may also be due heterogeneity of cell lines studied. PBMC from healthy controls were not in cell cycle. G2/M profiles and growth curves showed B-lymphoid cells just reaching plateau after 72 hour culture while myeloid cells were declining. IC50 values from cytotoxicity assay revealed myeloid cell lines had an average 13-fold higher drug resistance to epirubicin compared to B-lymphoid cell lines. Only CCL1, was expressed at least two-fold higher in myeloid compared to B-lymphoid cell lines. In contrast, MTRNR2, EEF1A1, PTMA, HLA-DR, C6orf115, PBX3, ENPP4, SELL, and IL3Ra were expressed more than 2-fold higher in B-lymphoid compared to myeloid cell lines studied here. Conclusion: Thus, B-lymphoid leukaemia cell lines here exhibited active, proliferating characteristics closer to GP genes. Higher expression of several genes in B-lymphoid compared to myeloid leukaemia cell lines may be useful markers to study biological differences including drug resistance between lineages.
Neoplasms

Introduction:Mesenchymal stem cells (MSCs) hold a great therapeutic potential for regenerative medicine and tissue engineering due to inherent immunomodulatory and reparative properties. Hence, it necessitates a readily available supplyof MSCs to meet the clinical demands adequately. Although, a human placenta can produce MSCs, the in vitro culture-mediated cellular senescence often affect the quality of cell product. Thus, the current study has explored the feasibility of basic fibroblast growth factor (bFGF) to enhance the growth of placenta-derived MSCs (PLC-MSCs). Methods:The basic fibroblast growth factor (bFGF) was supplemented to optimise the growth of MSCs. The effects of bFGF on morphology, growth kinetics and cytokine secretion of PLC-MSCs were assessed. Results: The bFGF supplementation increased the proliferation of PLC-MSCs in a dose-dependent manner and 40 ng/ml showed a high trophism effect on PLC-MSC’s growth. In the presence of bFGF, PLC-MSCs acquired a small and well-defined morphology that reflect an active proliferative status. BFGF has induced PLC-MSCs to achieve a shorter doubling time (45 hrs) as compared to the non-supplemented PLC-MSCs culture (81 hrs). Furthermore, bFGF impelled PLC-MSCs into cell cycle machinery where a substantial fraction of cells was driven to S and G2/M phases. Amongst, 36 screened cytokines, bFGF had only altered the secretion of IL-8, IL-6, TNFR1, MMP3 and VEGF. Conclusion:The present study showed that bFGF supplementation promotes the growth of PLC-MSCs without significantly deviating from the standard criteria of MSCs. Thus, bFGF could be considered as a potential mitogen to facilitate the large-scale production of PLC-MSCs.
Mesenchymal Stromal Cells

Osteolytic disease of the bones have a myriad range of aetiology. One rare cause is Gorham’s disease or disappearing bone disease. This disease is a diagnosis by exclusion using correlation made with clinical presentation, radiological findings and histopathological confirmation. Although many different therapies have been advocated, none have been successful in fully controlling this disease. We present a case that was detected in a Chinese lady using diagnostic imaging, confirmed with computed tomography guided biopsy and successfully treated with joint reconstruction using endoprosthesis.
Osteolysis, Essential

Thrombosis is one of the causes of morbidity and mortality in women of reproductive age group. Thrombosis at unusual sites may pose diagnostic and management dilemma for health care personnel. Teamwork and good communication provide the best modalities for maximum benefits to patients. Here with, we presented case a series of thrombosis at unusual sites seen and managed in our clinic. A 35 year-old Malay lady presented with left hemiparesis while she was on oestrogen based combined contraception pills (C-OCP). Imaging studies showed extensive venous thrombosis with bilateral acute cortical infarct. Thrombophilia screening of antiphospholipid syndrome were negative. She was put on anticoagulant and stopped 2 years after the incident. A 40 year-old Malay lady presented with abdominal discomfort, lethargy and massive splenomegaly. Bone marrow and trephine examination revealed primary myelofibrosis with positive JAK2617F. Imaging study showed chronic portal vein thrombosis with portal vein hypertension, complicated by gastro-oesophageal varices. She was put on hydroxyurea and later started on ruxolitinib with banding done over her gastro-oesophageal varices. A 26 year-old Malay lady presented with serositis, mouth ulcer and anaemia symptoms. Laboratory studies were positive for systemic lupus erythematosus and negative for antiphospholipid study. Imaging study showed long segment thrombosis of right internal jugular vein with surrounding subcutaneous oedema. She is currently stable on anticoagulants and steroid. Teamwork and holistic approach is practiced in the investigation and management to provide maximum benefits for patients.
Thrombosis