Aims: Particulate methane monooxygenase (pMMO) is an integral membrane protein that converts methane to methanol as the first step in the metabolic pathway of methanotroph bacteria. Methanotroph have a slow growth rate that make researcher have to develop an alternative approach by expressing the pMMO genes in Escherichia coli. However, it was very difficult to express all the pMMO encoded genes in E. coli and it is suspected that the protein might be toxic to E. coli. Therefore, this research tried another approach by expressing the active site of pMMO enzyme; cupredoxin domain of pmoB subunit encoded by spmoB gene. Methodology and results: The spmoB gene from Methylococcus capsulatus (Bath) was expressed in E. coli BL21 (DE3) under T7 promoter and pET15b as the expression vector. Several modifications were made so this gene would be expressed in the cytoplasm. Expression analysis with SDS-PAGE showed that overexpression of this gene could be done at several concentrations of IPTG and incubation temperature. The spmoB gene expression produced a recombinant protein with a size approximately 38.9 kDa. Assay of spmoB protein activity showed that the amount of methanol accumulated during methane oxidation by the recombinant strain was 0.114 mmol/mL culture.h. Conclusion, significance and impact study: We successfully expressed spmoB gene in E. coli BL21 (DE3) without high production of toxic compounds and it has methane oxidation activity. This result allowed further characterization of its potential applications.
Escherichia coli

Introduction Motor vehicle transport is common everywhere and in our local setting, most travel using their own cars. Resumption of driving after acquired brain injury (ABI) has been shown to affect subsequent community integration. Forty four to 66% of ABI survivors return to driving according to studies elsewhere. This study assesses the number of patients who resumed driving after ABI in Brunei Darussalam. Materials and Methods Brunei Darussalam citizens after ABI who were above 18 years old and had received occupational therapy in RIPAS Hospital, Brunei Darussalam between June 1, 2008 and May 31, 2009 were retrospectively identified and studied. Data was collected from the Department of Occupational Therapy records supplemented by telephone calls to patients or their relatives. Results Of 111 patients included in the study, 43 patients did not drive prior to ABI, since they were either older in age or never had a driving license. Of those with driving licenses (68) prior to ABI, 15 (22%) resumed driving: traumatic brain injury (n=1/5, 20%), brain tumour (n=1/4, 25%), brain infection (n=1/1, 100%) and for stroke (n=12/58, (20.7%). Gender and age groups were not significant predictors of return to driving. Conclusion Only 22% of Bruneians after ABI resumed driving and this is lower than studies elsewhere. Reasons considered include severity of injury and cultural factors which all require further study.

A gentleman in his forties presented with drooping of his right eye (Panel A). A test was done and there was improvement (Panel B). What is the diagnosis and what ‘test’ was done? Answer: refer to page 155

(Refer to page 133) Answer: Ocular myasthenia gravis and an ‘ice pack test’, a two minute application of ice (wrapped in a clean towel or other protective covering) to the affected levator muscle. Myasthenia gravis (MG) is an acquired autoimmune disease caused by antibodies to the acetylcholine receptors at the neuromuscular junction of skeletal muscle. The resultant defect in neuromuscular transmission causes muscle fatiguability and weakness. Weakness may localise purely to eye muscles termed ocular myasthenia or may affect other skeletal muscle termed generalised MG. About 50 to 70% of ocular myasthenia will eventually develop generalised MG.

Diabetic retinopathy (DR) may result in progressive visual impairment and blindness which affects the diabetic patients‟ morbidity and quality of life significantly. The objective of this study was to determine the proportions and prognostic factors of DR severity among Type 2 diabetes mellitus patients. A cross sectional study was conducted at the Ophthalmology Clinic in a tertiary hospital using the medical record database from 2005 to 2011. A total of 216DR patients were randomly selected. The study outcome was DR severity which was classified into four grading based on the International Clinical Diabetic Retinopathy Scale. Descriptive statistics and Ordinal Logistic Regression were applied using Stata SE/11. The mean (SD) age of DR in this study was 56.06 (9.98) years old with almost equal sex ratio. Proportion of mild non proliferative diabetic retinopathy (NPDR) was 48.6% (95% CI: 40.0, 60.0), moderate NPDR was 28.2% (95%CI: 22.0, 40.0), severe NPDR was 6.9% (95% CI: 4.0, 11.0) and proliferative diabetic retinopathy (PDR) was 16.2% (95%CI: 10.0, 22.0). Diabetic patients that suffered from diabetes mellitus for more than 10 years, having nephropathy, every increased of HbA1c by 1% and higher total cholesterol were at increased risk of worsening DR.

Aims: An ecological study was conducted to investigate the diversity pattern of the lactic acid bacteria in the gastrointestinal tract of Cemani chicken, a native Indonesian chicken, using a molecular approach based on 16S rRNA genes. Methodology and results: Digesta samples of seven chickens were collected for terminal restriction fragment length polymorphism (T-RFLP) analysis. The molecular diversity of lactic acid bacteria in crop, ventriculus, ileum and cecum were determined. The results showed that microbial composition of lactic acid bacteria in cecum was relatively different with other upper gastrointestinal tract. Lactic acid bacteria phylotypes and diversity in ileum were higher than those in the crop, ventriculus, and cecum. Conclusion, significance and impact of study: We confirmed that cecum of native chicken has a different environment as compared to other gastrointestinal regions showing the lowest value of the Sorensen’s index. This first report of LAB diversity pattern in Cemani chicken contributes a more comprehensive understanding of the microbial ecology in the chicken.
Gastrointestinal Tract

Introduction: The Hedgehog (Hh) signalling pathway is a developmental signalling pathway involved in normal mammalian developmental and homeostasis of adult renewable tissues. In most adult tissues, this pathway remains silent and previous studies have shown that constitutive activation of Hedgehog signalling pathway leads to various types of malignancies including medulloblastomas, basal cell carcinoma, gastrointestinal, breast and prostate cancer. The purpose of this study was to investigate the immunohistochemical expression of Hedgehog pathway proteins in Diffuse Large B-cell Lymphoma and determine their association with overall survival (OS). Methods: Positive control using normal tonsils were included in each batch of immunohistochemical staining procedure. Results: PTCH1 proteins were highly expressed in DLBCL and showed strong staining intensity in 107 (100%) cases and SMO proteins were expressed in 105 (98.1%) cases. PTCH1 proteins were localised in the nucleus of tumour cells, whereas SMO proteins were mainly localised in the cytoplasm of tumour cells. Positive expression of PTCH1 and SMO proteins and overall survival of DLBCL patients were correlated with age, gender, race and tumour location. There was no significant correlation between the expression of these two proteins with any of the parameters. PTCH1 expression showed significant association with SMO expression (P=0.03). Conclusions: Our findings suggest that high expression of both PTCH1 and SMO may be important in the pathogenesis of DLBCL. However, additional mechanisms that may contribute to the activation of HH signalling in DLBCL needs to be further explored.

An investigation on ablution workstations design for person with disabilities has been carried out in this study. The focused were on wheelchair user or person with lower-limb impairment in Malaysia. The objective of this research is to determine the most suitable ablution workstation for person with disabilities. One hundred wheelchair users which include 74 males and 26 females participated in the study. The findings highlighted that the most suitable and preferred ablution workstation for person with disabilities is a separate design for washing hands and legs. The results also indicated that the noteworthy body parts that should be enhanced are arms, neck, trunk, and legs. Therefore, it can be concluded that it is important to design an ablution workstation with correct anthropometric dimensions referring to the population under investigation taking into account their preferences and disabilities limitation. Thus, the innovation will bring equality between people with disabilities and physically normal people in future facilities design. The workstation will become one of the worthy social contributions to the population of Muslim disabilities especially in Malaysia.

Whiplash injury due to low severity vehicles crash is a global problem. The injury has long-term clinical and biomechanical implications. Since the mid-1960s, injury statistics have continuously revealed that females face a higher risk of suffering the injury category compare to males. Besides, in a frontal crash, the injury measures from the adult rear dummies were mainly higher than the same size dummies located in driver and front occupant seat. However, most regulations and user crash tests have focused on vehicle drivers and front-seat passenger due to high occupancy and mortality rates in the front seat. In this paper, mechanisms of whiplash injury were reviewed to contribute a further inclusive understanding of human impact reaction, variability quantification, validation, and prevention. The objective of this study is to develop a new design of head restraint (HR) for car rear occupants. In order to raise consideration whiplash injury and prevention mechanisms, impacts are simulated with computer modelling (Ls-Dyna simulation) and validated using Matlab. Therefore, a review of these injury mechanisms indicates the development of new anti-whiplash technology in the automotive safety area is necessary.