Objective: To establish a polymerase chain reaction (PCR) technique based on cytochrome b (cytb) gene of mitochondria DNA (mtDNA) for blood meal identification. Methods: The PCR technique was established based on published information and validated using blood sample of laboratory animals of which their whole gene sequences are available in GenBank. PCR was next performed to compile gene sequences of different species of wild rodents. The primers used were complementary to the conserved region of the cytb gene of vertebrate’s mtDNA. A total of 100 blood samples, both from laboratory animals and wild rodents were collected and analyzed. The obtained unknown sequences were compared with those in the GenBank database using BLAST program to identify the vertebrate animal species. Results: Gene sequences of 11 species of wild animals caught in 9 localities of Peninsular Malaysia were compiled using the established PCR. The animals involved were Rattus (rattus) tanezumi, Rattus tiomanicus, Leopoldamys sabanus,Tupaia glis, Tupaia minor, Niviventor cremoriventor, Rhinosciurus laticaudatus, Callosciurus caniseps, Sundamys muelleri, Rattus rajah and Maxomys whiteheadi. The BLAST results confirmed the host with exact or nearly exact matches (>89% identity). Ten new gene sequences have been deposited in GenBank database since September 2010. Conclusions: This study indicates that the PCR direct sequencing system using universal primer sets for vertebrate cytb gene is a promising technique for blood meal identification.

The purpose of this study is to determine the standard value of corneal thickness (CCT) and its relationship with the degree of myopia amongst one sample of myopic students in Malaysia. Eighty-four myopic subjects (168 eyes) were examined in this study. Ocular examination included subjective refraction, keratometry (Shin-Nippon, Japan) and optical pachometry (Sonogage INC). Corneal thickness was measured at five different locations i.e. central, superior, inferior, nasal and temporal of the cornea. However, only association of CCT with myopia is reported in this article. Mean age of subjects was 21.42 ± 1.47 years old (20.34 ± 1.15; 22.46 ± 0.88). Mean refractive error in spherical equivalent for the whole sample was –3.50 ± 2.10 DS. Mean corneal curvature for the all subjects was 7.74 ± 0.52 mm and mean value of CCT was 594.62 ± 66.51 mm. Significant differences (p < 0.05) were found between mean CCT of low myopes (-4.00DS and below) and higher ones (-4.25DS and above), but not with mean of keratometer readings. Poor correlations were obtained between refractive error and CCT (r = 0.125, p > 0.05) and between refractive error and corneal curvature (r = 0.24, p > 0.05). It was concluded from this study that high myopic subjects has significantly thinner CCT than lower ones. Measurement of CCT should be considered when undertaking procedures such as LASIK and orthokeratology.

There is a need to design and develop an affordable motorized vehicle for better mobility among physical disabilities. This study aimed is to initiate appropriate and affordable materials to develop an accessible motorized wheelchair. It includes the feasibility study of the existing used wheelchair, the system used and the characteristic of the physical disabilities symptom. Through the observation, an existing motorized wheelchair uses stainless steel as the main structure, manipulating the shelves heavy-duty accessories, electrical component, heavy-duty battery and car pulley system in which the weight accumulates approximately 100 kg. The heavyweight wheelchair cannot be folded and put inside the car. It also needs to have a special carrier to carry the wheelchair. The field research has also been conducted to evaluate the existing wheelchair system in the market with various systems used, components, and motors material used in the area of technical, usability and comfort before start to redesigning the wheelchair. The results of the study would include the background needs of the characteristic of physical disabilities. The available high-grade aluminium materials will be used to support the mobility and their relevant properties. The design will be done all the way and the specification will be used as an instrument data for the motorized vehicle. The selection of design features, documentation of design procedures and results will be used to create a P.O.C (Prove of Concept) prototype for the meant vehicle. The user will use the preliminary prototyping test and the data (result) will be used to improvise the wheelchair prototype. The second improvised prototype will undergo the second user testing in the area of reliability, stabilization of system, and ease of usage. The results expected will be observed and tested by the user to lead towards the recommendation of the motorized wheelchair design.

Cardio-pulmonary Resuscitation (CPR) is important and should be mastered by House Officers (HO). House officers who have just completed their studies are assigned to acute medical and surgical wards. If a patient in the ward has a cardiac arrest (CA), these doctors are usually the first to attend. Therefore an HO must be confident with CPR skills. They must be competent in performing CPR. The authors assessed 26 new HOs from Universiti Kebangsaan Malaysia Hospital (HUKM) with respect to their self-perception about CPR skills, confidence level in performing CPR and knowledge in performing CPR. Knowledge was assessed by a questionnaire. We found that 16 of 26 (61.5%) assessed themselves to have inadequate knowledge and 46.2% had no confidence in performing CPR. The mean score of the written test was 5.7 ± 1.8. Seven out of 26 (27.0%) HOs had incorrect hand placement position for CPR. Only 4 and 9 out of 26 HOs had their sternal paddle and cardiac apex paddle positions correctly placed respectively. In conclusion, knowledge, perception of skills and confidence levels of HOs on CPR are inadequate and need further assessment and improvement. Medical schools need to review their CPR curriculum in order to prepare HOs adequately to work in emergency situations.

beta-Lactamases have been identified as the major cause of antimicrobial resistance to beta-lactam antibiotics in Escherichia coli. The activities of ampicillin-sulbactam and amoxicillin-clavulanate as well as a range of beta-lactam antibiotics were studied with 87 clinical E. coli isolates from patients of the University Malaya Medical Center using the disc diffusion technique. Susceptible, intermediate and resistant categories were established based on the diameter of zones of inhibition set by the National Committee for Clinical Laboratory Standards (NCCLS). The isolates were then classified into 6 phenotypes according to the criteria stated in the methodology: S (susceptible to all beta-lactams); TL (resistant to aminopenicillins; amoxicillin-clavulanate susceptible and susceptible or intermediate to ampicillin-sulbactam); TI (resistant to aminopenicillins and ampicillin-sulbactam; susceptible to amoxicilin-clavulanate); TH-IRT (resistant to aminopenicillins; intermediate or resistant to amoxicillin-clavulanate; resistant to ampicillin-sulbactam); ESBL (resistant to aminopenicillins and oxyimino cephalosporins; positive results with the double-disc diffusion test); and CP (resistant to aminopenicillins, beta-lactam-beta-lactamase inhibitor combinations, oxyimino cephalosporins and cephamycins). Results showed that the TL phenotype was the commonest (40.2% of the isolates) followed by S (31%), TH-IRT (16.1%), ESBL and CP (3.4% each) and TI (2.3%). One isolate showed both ESBL and CP phenotypes while two isolates were classified as inconclusive. Representatives from each phenotype were further analysed for the presence of beta-lactamases which revealed a predominance of TEM and SHV enzyme producers. PCR-SSCP analysis of the SHV gene from all the ESBL and CP isolates revealed the predominance of SHV 5-type enzyme which was concurrent with our previous studies.
Phenotype ; Extended-spectrum beta lactamase ; sultamicillin ; Escherichia coli ; Ceruloplasmin

BACKGROUND: The urinary tract can be affected by both congenital abnormalities as well as acquired disorders, such ascancer, trauma, infection, inflammation, and iatrogenic injuries, all of which may lead to organ damage requiring eventualreconstruction. As a gold standard, gastrointestinal segment is used for urinary bladder reconstruction. However, one majorproblem is that while bladder tissue prevents reabsorption of specific solutes, gastrointestinal tissue actually absorbs them.Therefore, tissue engineering approach had been attempted to provide an alternative tissue graft for urinary bladderreconstruction. METHODS: Human adipose-derived stem cells isolated from fat tissues were differentiated into smooth muscle cells andthen seeded onto a triple-layered PLGA sheet to form a bladder construct. Adult athymic rats underwent subtotal urinarybladder resection and were divided into three treatment groups (n = 3): Group 1 (‘‘sham’’) underwent anastomosis of theremaining basal region, Group 2 underwent reconstruction with the cell-free scaffold, and Group 3 underwent reconstructionwith the tissue-engineered bladder construct. Animals were monitored on a daily basis and euthanisation wasperformed whenever a decline in animal health was detected. RESULTS: All animals in Groups 1, 2 and 3 survived for at least 7 days and were followed up to a maximum of 12 weekspost-operation. It was found that by Day 14, substantial ingrowth of smooth muscle and urothelial cells had occurred inGroup 2 and 3. In the long-term follow up of group 3 (tissue-engineered bladder construct group), it was found that theurinary bladder wall was completely regenerated and bladder function was fully restored. Urodynamic and radiologicalevaluations of the reconstructed bladder showed a return to normal bladder volume and function.Histological analysisrevealed the presence of three muscular layers and a urothelium similar to that of a normal bladder. Immunohistochemicalstaining using human-specific myocyte markers (myosin heavy chain and smoothelin) confirmed the incorporation of theseeded cells in the newly regenerated muscular layers. CONCLUSION Implantation of PLGA construct seeded with smooth muscle cells derived from human adipose stemcells can lead to regeneration of the muscular layers and urothelial ingrowth, leading to formation of a completelyfunctional urinary bladder.

BACKGROUND: The urinary tract can be affected by both congenital abnormalities as well as acquired disorders, such ascancer, trauma, infection, inflammation, and iatrogenic injuries, all of which may lead to organ damage requiring eventualreconstruction. As a gold standard, gastrointestinal segment is used for urinary bladder reconstruction. However, one majorproblem is that while bladder tissue prevents reabsorption of specific solutes, gastrointestinal tissue actually absorbs them.Therefore, tissue engineering approach had been attempted to provide an alternative tissue graft for urinary bladderreconstruction. METHODS: Human adipose-derived stem cells isolated from fat tissues were differentiated into smooth muscle cells andthen seeded onto a triple-layered PLGA sheet to form a bladder construct. Adult athymic rats underwent subtotal urinarybladder resection and were divided into three treatment groups (n = 3): Group 1 (‘‘sham’’) underwent anastomosis of theremaining basal region, Group 2 underwent reconstruction with the cell-free scaffold, and Group 3 underwent reconstructionwith the tissue-engineered bladder construct. Animals were monitored on a daily basis and euthanisation wasperformed whenever a decline in animal health was detected. RESULTS: All animals in Groups 1, 2 and 3 survived for at least 7 days and were followed up to a maximum of 12 weekspost-operation. It was found that by Day 14, substantial ingrowth of smooth muscle and urothelial cells had occurred inGroup 2 and 3. In the long-term follow up of group 3 (tissue-engineered bladder construct group), it was found that theurinary bladder wall was completely regenerated and bladder function was fully restored. Urodynamic and radiologicalevaluations of the reconstructed bladder showed a return to normal bladder volume and function.Histological analysisrevealed the presence of three muscular layers and a urothelium similar to that of a normal bladder. Immunohistochemicalstaining using human-specific myocyte markers (myosin heavy chain and smoothelin) confirmed the incorporation of theseeded cells in the newly regenerated muscular layers. CONCLUSION Implantation of PLGA construct seeded with smooth muscle cells derived from human adipose stemcells can lead to regeneration of the muscular layers and urothelial ingrowth, leading to formation of a completelyfunctional urinary bladder.

Background: Adipose tissue provides an abundant source of multipotent cells, which represent a source of cell-based regeneration strategies for urinary bladder smooth muscle repair. Our objective was to confirm that adipose-derived stem cells (ADSCs) can be differentiated into smooth muscle cells. Methods: In this study, adipose tissue samples were digested with 0.075% collagenase, and the resulting ADSCs were cultured and expanded in vitro. ADSCs at passage two were differentiated by incubation in smooth muscle inductive media (SMIM) consisting of MCDB I31 medium, 1% FBS, and 100 U/mL heparin for three and six weeks. ADSCs in non-inductive media were used as controls. Characterisation was performed by cell morphology and gene and protein expression. Result: The differentiated cells became elongated and spindle shaped, and towards the end of six weeks, sporadic cell aggregation appeared that is typical of smooth muscle cell culture. Smooth muscle markers (i.e. alpha smooth muscle actin (ASMA), calponin, and myosin heavy chain (MHC)) were used to study gene expression. Expression of these genes was detected by PCR after three and six weeks of differentiation. At the protein expression level, ASMA, MHC, and smoothelin were expressed after six weeks of differentiation. However, only ASMA and smoothelin were expressed after three weeks of differentiation. Conclusion: Adipose tissue provides a possible source of smooth muscle precursor cells that possess the potential capability of smooth muscle differentiation. This represents a promising alternative for urinary bladder smooth muscle repair.
Adipose Tissue ; Stem Cells ; Muscle Cells ; Regeneration ; Urinary Bladder

Introduction: Hyperhidrosis is a disorder of excessive and uncontrollable sweating beyond the body’s physiological needs. It can be categorised into primary or secondary hyperhidrosis based on its aetiology. Detailed history review including onset of symptoms, laterality of disease and family history are crucial which may suggest primary hyperhidrosis. Secondary causes such as neurological diseases, endocrine disorders, haematological malignancies, neuroendocrine tumours and drugs should be adequately examined and investigated prior to deciding on further management. The diagnosis of primary hyperhidrosis should only be made only after excluding secondary causes. Hyperhidrosis is a troublesome disorder that often results in social, professional, and psychological distress in sufferers. It remains, however, a treatment dilemma among some healthcare providers in this region. Methods: The medical records and clinical outcomes of 35 patients who underwent endoscopic thoracic sympathectomy for primary hyperhidrosis from 2008 to 2018 in Department of Cardiothoracic Surgery were reviewed. Results: The mean age of the patients was 27±10.1years, with male and female distribution of 18 and 17, respectively. Fifty-one percent of patients complained of palmar hyperhidrosis, while 35% of them had concurrent palmaraxillary and 14% had palmar-plantar-axillary hyperhidrosis. Our data showed that 77% (n=27) of patients were not investigated for secondary causes of hyperhidrosis, and they were not counselled on the non-surgical therapies. All patients underwent single-staged bilateral endoscopic thoracic sympathectomy. There was resolution of symptoms in all 35 (100%) patients with palmar hyperhidrosis, 13(76%) patients with axillary hyperhidrosis and only 2 (50%) patients with plantar hyperhidrosis. Postoperatively 34.3% (n=12) of patients reported compensatory hyperhidrosis. There were no other complications such as pneumothorax, chylothorax, haemothorax and Horner’s Syndrome. Conclusion: Clinical evaluation of hyperhidrosis in local context has not been well described, which may inadvertently result in the delay of appropriate management, causing significant social and emotional embarrassment and impair the quality of life of the subjects. Detailed clinical assessment and appropriate timely treatment, be it surgical or non-surgical therapies, are crucial in managing this uncommon yet distressing disease.

BACKGROUND: Urinary tract is subjected to a variety of disorders such as urethral stricture, which often develops as a result of scarring process. Urethral stricture can be treated by urethral dilation and urethrotomy; but in cases of long urethral strictures, substitution urethroplasty with genital skin and buccal mucosa grafts is the only option. However a number of complications such as infection as a result of hair growth in neo-urethra, and stone formation restrict the application of those grafts. Therefore, tissue engineering techniques recently emerged as an alternative approach, aiming to overcome those restrictions. The aim of this review is to provide a comprehensive coverage on the strategies employed and the translational status of urethral tissue engineering over the past years and to propose a combinatory strategy for the future of urethral tissue engineering. METHODS: Data collection was based on the key articles published in English language in years between 2006 and 2018 using the searching terms of urethral stricture and tissue engineering on PubMed database. RESULTS: Differentiation of mesenchymal stem cells into urothelial and smooth muscle cells to be used for urologic application does not offer any advantage over autologous urothelial and smooth muscle cells. Among studied scaffolds, synthetic scaffolds with proper porosity and mechanical strength is the best option to be used for urethral tissue engineering. CONCLUSION: Hypoxia-preconditioned mesenchymal stem cells in combination with autologous cells seeded on a prevascularized synthetic and biodegradable scaffold can be said to be the best combinatory strategy in engineering of human urethra.
Cicatrix ; Data Collection ; Hair ; Humans ; Mesenchymal Stromal Cells ; Mouth Mucosa ; Myocytes, Smooth Muscle ; Porosity ; Skin ; Tissue Engineering ; Transplants ; Urethra ; Urethral Stricture ; Urinary Tract