The Importance of Molecular Biological Analysis for the Laboratory Diagnostic of Homozygous Haemoglobin Malay.

Haemoglobin (Hb) Malay is variant haemoglobin with a ß++ thalassemia phenotype. The prevalence of Hb Malay in the Malaysian population was 5.5%. We describe a 58-year-old male who presented with symptomatic anaemia to the Hospital Universiti Sains Malaysia. Further history revealed that the patient had anaemia since the age of 28, and on regular follow-up at other hospital. Physical examination revealed pallor, jaundice and hepatosplenomegaly. The full blood count and peripheral blood smear showed hypochromic microcytic anaemia with anisopoikilocytosis, and many target cells. High-performance liquid chromatography results showed a ß thalassemia trait. However, the diagnosis does not alight with the patient's condition. Bone marrow aspirate was completed and showed reactive changes and erythroid hyperplasia. A molecular test was then performed for ß globin gene mutation detection using Multiplex Amplification Refractory Mutation System (M-ARMS) PCR method. This revealed the result as homozygous codon 19 mutation or Hb Malay. Therefore, in this case report we would like to highlight the laboratory approaches, the challenges faced by the usual haematological investigations and the importance role of molecular testing in the diagnosis of severe anaemia.

A rare case of compound heterozygous Southeast Asian double α-globin gene deletion and Haemoglobin Quong Sze in a Malay proband.

INTRODUCTION: Haemoglobin (Hb) Quong Sze is a non-deletional α-thalassaemia subtype that occurs due to missense mutation at codon 125 of the HBA2 gene. Interaction between Hb QS with Southeast Asian double α-globin gene deletion results in non-deletional HbH disease, which is more severe than deletional HbH. CASE REPORT: A 3-month-old baby boy was presented with neonatal anaemia and mild hepatomegaly. Full blood count revealed severe hypochromic microcytic anaemia. There was an abundance of HbH inclusion bodies in his red blood cells. High-performance liquid chromatography showed a reduced HbA2 level with the presence of pre-run peak. Capillary electrophoresis showed the presence of HbH and Hb Barts. Molecular analysis found a common α0-thalassaemia (--SEA) in one allele and mutation in codon 125 in the other allele. DISCUSSION: Non-deletional HbH disease due to a combination of deletional and non-deletional mutations may present with severe clinical manifestations than those with deletion mutations, which warrants accurate diagnosis using molecular techniques.

A Rare Case of an Infant with Left Hemiparesis: A Case Report of Bilateral Open-lip Schizencephaly.

Schizencephaly is a very rare congenital birth defect. It is characterized by a cortical brain malformation that manifests as a grey-matter-lined cleft extending from the ependyma to the pia mater. It is a rare condition, and few cases have been reported in the literature. The exact cause is unknown. Herein, we report a case of an infant presenting with left side hemiparesis. The CT scan of her brain revealed right fronto-temporal and left parieto-temporal open-lip schizencephaly; thus, urgent referral to a pediatric neurologist was made for early intervention.

Young Lady with Bilateral Yellowish Lesions on Her Eyelids.

A 26-year-old woman with an unknown medical illness presented with yellowish skin lesions around both eyes, visible for the past 4 years. The lesions were neither itchy nor tender and had increased gradually in size. She had neither constitutional nor hyperthyroidism symptoms. She is the youngest out of four siblings, and none of her family members have cardiovascular disease or similar problems. Upon examination, there were bilateral yellowish plaques over the periorbital region. Her body mass index (BMI) was 23.8 kg/m2. Her vital signs were all normal at every visit.

Coracoid Process Morphology using 3D-CT Imaging in a Malaysian Population.

Introduction: The aims of this study are to define the coracoid process anatomy in a Malaysian population, carried out on patients in Hospital Serdang with specific emphasis on the dimension of the base of coracoid process which is important in coraco-acromial (CC) ligament reconstruction, to define the average amount of bone available for use in coracoid transfer, and to compare the size of coracoid process based on gender and race, and with findings in previous studies. Materials and Methods: Fifteen pairs of computed tomography (CT) based 3-dimensional models of shoulders of patients aged between 20 to 60 years old were examined. The mean dimensions of coracoid were measured and compared with regards to gender and race. The data were also compared to previously published studies. Results: The mean length of the coracoid process was 37.94 ± 4.30 mm. Male subjects were found to have larger-sized coracoids in all dimensions as compared to female subjects. The mean tip of coracoid dimension overall was 19.99 + 1.93mm length × 10.03 + 1.48mm height × 11.63 + 2.12mm width. The mean base of coracoid dimension was 18.96 + 3.71mm length × 13.84 + 1.76mm width. No significant differences were observed with regards to racial denomination. The overall coracoid size measurements were found to be smaller compared to previous studies done on the Western population. Conclusion: This study may suggest that Malaysians have smaller coracoid dimension compared to Caucasians. The findings further suggest that the incidence of coracoid fracture and implants pull out in Malaysian subjects may be higher.

An association of myasthenia gravis with Hashimoto's thyroiditis in a patient with a multinodular goitre.

INTRODUCTION: The association of myasthenia gravis (MG) with other autoimmune diseases including autoimmune thyroid disease (ATD) is well recognised, although rare. The occurrence of both diseases can occur in two ways: either disease preceding the other, or concurrently. The presentation of MG in association with ATD can range from ocular to generalised disease. CASE SUMMARY: A 26-year-old Malay female with persistent hyperthyroidism secondary to Hashimoto's thyroiditis in multinodular goitre was diagnosed with generalised MG after 2 years. She presented with right eye ptosis (ocular) and difficulty in swallowing and chewing (bulbar). The diagnosis of MG was confirmed by fatigability testing, electromyography and the presence of AChR antibodies. Her symptoms showed improvement with pyridostigmine (Mestinon) 60 mg 6-hourly. Her antithyroid drug was tapered down according to her thyroid function test. Throughout a year of follow-ups, her hyperthyroidism and fatigability symptoms improved with treatment. She was later counselled for total thyroidectomy and thymectomy. CONCLUSION: Myasthenia gravis and hyperthyroidism may present with similar symptoms such as dysphagia due to neuromuscular weakness or fatigue. When the diseases occur together, one of the diagnoses may be missed. Therefore, the occurrence of new symptoms in a patient with underlying ATD should should trigger the early identification of other autoimmune diseases by primary care doctors.

Coracoid Process Morphology using 3D-CT Imaging in a Malaysian Population

Introduction: The aims of this study are to define the coracoid process anatomy in a Malaysian population, carried out on patients in Hospital Serdang with specific emphasis on the dimension of the base of coracoid process which is important in coraco-acromial (CC) ligament reconstruction, to define the average amount of bone available for use in coracoid transfer, and to compare the size of coracoid process based on gender and race, and with findings in previous studies. Materials and Methods: Fifteen pairs of computed tomography (CT) based 3-dimensional models of shoulders of patients aged between 20 to 60 years old were examined. The mean dimensions of coracoid were measured and compared with regards to gender and race. The data were also compared to previously published studies. Results: The mean length of the coracoid process was 37.94 ± 4.30 mm. Male subjects were found to have larger-sized coracoids in all dimensions as compared to female subjects. The mean tip of coracoid dimension overall was 19.99 + 1.93mm length x 10.03 + 1.48mm height x 11.63 + 2.12mm width. The mean base of coracoid dimension was 18.96 + 3.71mm length x 13.84 + 1.76mm width. No significant differences were observed with regards to racial denomination. The overall coracoid size measurements were found to be smaller compared to previous studies done on the Western population. Conclusion: This study may suggest that Malaysians have smaller coracoid dimension compared to Caucasians. The findings further suggest that the incidence of coracoid fracture and implants pull out in Malaysian subjects may be higher.

Potential use of cord blood for Hb E hemoglobinopathy screening programme using capillary electrophoresis.

BACKGROUND: Thalassemia and hemoglobinopathies are inherited red blood cell disorders found worldwide. Hemoglobin (Hb) E disorder is one of the hemoglobinopathies known to have the high prevalence in South East Asia. Most of transfusion-dependent thalassemias were genotypically compound heterozygous Hb E/ ß-thalassemia. In Malaysia, the national screening program for thalassemia was implemented for early pregnancy or secondary school girls; however many participants do not turn-up and missed the screening test. Screening for thalassemia using samples from cord blood is an alternative choice as it is a readily available source of blood and hence early detection of the disease. The purpose of this study was to determine the potential use of cord blood for the screening of HbE hemoglobinopathy by using capillary electrophoresis (CE). METHODS: Cord blood samples were collected from 300 newborns of healthy mothers. Hematological parameters were determined and hemoglobin quantitation for all cord blood samples were performed using capillary electrophoresis system (CES) and high performance liquid chromatography (HPLC). RESULTS: Majority of cord blood samples (63%) revealed Hb AF followed by Hb AFA2 (20%). Hb AFE was detected in 10.7% with the mean value of Hb E ranging from 2.3%-11.1%. CONCLUSION: Hemoglobin E was detected in cord blood using capillary electrophoresis system. It can be recommended in areas where Hb E/ß is prevalent. Implementation of a screening strategy using CE on cord blood sampling will identify the disease early. With regular follow-up on these patients, the status of their disease can be determined earlier and appropriate management implemented.

Microsurgical skill acquisition in a one-day introductory course with performance evaluation using software-assisted scoring system.

BACKGROUND: Recent emphasis on microsurgical skill acquisition at an earlier stage of plastic surgery training has seen a shift toward objective competence-based assessment. Yet no objective measures of spacing or alignment exist, with few validated models that assess ability. The authors propose a novel software analysis scoring system to objectively measure spacing, alignment and the overall improvement in a 1-day, introductory course setting. METHODS: Images of standard 4-mm latex strips that had been sutured by participants using the Microtrainer system were uploaded onto calibrated, online software. Sutures were analysed with regard to spacing, alignment and density. From these measurements, a total score was calculated, one on initial assessment at the course beginning (Score 1) and another on final assessment at the course end (Score 2), thereby facilitating measurement of the overall improvement. RESULTS: A total of 38 microsurgical anastomoses from 19 participants ranging from postgraduate years 1-7 were analysed. Seventeen participants had no previous experience of microsurgery. The mean average Score 1 of participants was -2 (range -12 to +22) and Score 2 was 22 (range +12 to +32), thus showing a significant improvement in candidate ability throughout the course of the day (p < 0.0001). CONCLUSIONS: Microtrainer system software analysis provides a novel, reliable, and consistent objective assessment for surgical trainees at all stages of training, without risk to patients. It has an associated cost for the initial setup, yet is timely, repeatable and can efficiently demonstrate progress in a 1-day course setting.

N,N-dimethylhexadecyl carboxymethyl chitosan as a potential carrier agent for rotenone.

In this study, an amphiphilic chitosan derivative namely N,N-dimethylhexadecyl carboxymethyl chitosan (DCMC) was synthesised and applied for the first time as a carrier agent for rotenone. The physical and chemical properties of DCMC were characterised by using Fourier Transform Infrared Spectrometer (FTIR), Proton Nuclear Magnetic Resonance Spectrometer ((1)H NMR), CHN-O Elemental Analyser, Thermogravimetric Analyser (TGA) and Differential Scanning Calorimeter (DSC). DCMC was soluble in acidic (except pH 4), neutral and basic media with percent of transmittance (%T) values ranged from 67.2 to 99.4%. The critical micelle concentration (CMC) was determined as 0.095mg/mL. Transmission Electron Microscopy (TEM) analysis confirmed that DCMC has formed self-aggregates and exhibited spherical shape with the size of 65.5-137.0nm. The encapsulation efficiency (EE) and loading capacity (LC) of DCMC micelles with different weight ratios (DCMC:rotenone; 5:1, 50:1 and 100:1) were determined by using High Performance Liquid Chromatography (HPLC). The weight ratio of 100:1 gave the best EE with the value of more than 95.0%. DCMC micelles performed an excellent ability to control the release of rotenone, of which 99.0% of rotenone was released within 48h. Overall, DCMC has several key features to be an effective carrier agent for pesticide formulations.

Immobilisation of Cu, Pb and Zn in Scrap Metal Yard Soil Using Selected Waste Materials.

Immobilisation of heavy metals in a 30-year old active scrap metal yard soil using three waste materials, namely coconut tree sawdust (CTS), sugarcane bagasse (SB) and eggshell (ES) was investigated. The contaminated soil was amended with amendments at application rates of 0 %, 1 % and 3 % (w/w). The effects of amendments on metal accumulation in water spinach (Ipomoea aquatica) and soil metal bioavailability were studied in a pot experiment. All amendments increased biomass yield and reduced metal accumulation in the plant shoots. The bioconcentration factor and translocation factor values of the metals were in the order of Zn > Cu > Pb. The addition of ES, an alternative source of calcium carbonate (CaCO3), has significantly increased soil pH and resulted in marked reduction in soil metal bioavailability. Therefore, CTS, SB and ES are promising low-cost immobilising agents to restore metal contaminated land.

A double mobility acetabular implant for primary hip arthroplasty in patients at high risk of dislocation.

INTRODUCTION: Dislocation following total hip replacement continues to be a problem for which no completely satisfactory solution has been found. Several methods have been proposed to reduce the incidence of hip dislocations with varying degrees of success, including elevated rim liners, constrained liners and large diameter bearings. We present our experience with the double mobility acetabular component in patients at high risk of instability. METHODS: This was a retrospective review of 65 primary total hip arthroplasties in 55 patients (15 men, 40 women), performed between October 2005 and November 2009. The majority (80%) of patients had at least two and 26% had at least three risk factors for instability. The mean age was 76 years (range: 44-92 years). The patients were followed up for a mean duration of 60 months (range: 36-85 months). RESULTS: Fourteen patients died and one was lost to follow-up, leaving fifty hips for final assessment. Until the final follow-up appointment, no patients had dislocation and none required revision surgery. The mean Oxford hip score improved from 45.0 to 26.5 (p<0.0001). The mean Merle d'Aubigné pain score improved from 1.4 to 4.9 (p<0.0001), the walking score from 2.3 to 3.1 (p<0.07) and the absolute hip function score from 5.4 to 10.8 (p<0.0001). There were no clinical or radiographic signs of loosening. CONCLUSIONS: The double mobility acetabular component was successful at preventing dislocation during early to medium-term follow-up. However, as data are still lacking with regard to polyethylene wear rates at the additional bearing surface, it would be prudent to restrict the use of this implant to selected patients at high risk of instability.

Initial evaluation of the training programme for health care professionals on the use of Malaysian clinical practice guidelines for management of dementia.

OBJECTIVE: The Malaysian Clinical Practice Guidelines (CPG) for Management of Dementia (second edition) was launched in April 2010 by the Ministry of Health Malaysia. A training programme for the management of dementia, involving all categories of staff working at primary and secondary centres, was implemented to ensure that care delivery for people with dementia was in accordance with the guidelines. The study aimed to look into improving knowledge and understanding of dementia following training, and to evaluate the effectiveness of the training programme using a clinical audit indicator recommended in the guidelines. METHODS: The study entailed 2 phases (at national and state levels). The first phase involved the CPG training programme run as a 1.5-day workshop, in which participants filled up pre- and post-workshop questionnaires. A second phase involved analysing all the referral letters to the memory clinic at the Hospital Sultan Ismail, Johor Bahru 1 year before and after the training programme. RESULTS: There was a significant improvement in knowledge about dementia and its management among the health care professionals following training. The mean percentage score for the pre-workshop test was 63% while for the post-workshop test it was 78%, giving a difference of 15%. Although there was an overall improvement in knowledge gain following training in both specialist and non-specialist groups, these differences were not statistically significant (t = 1.32; 95% confidence interval, -2.61 to 9.61; p = 0.25). The proportion of referrals with a possible diagnosis of dementia from primary clinic referrals to the memory clinic also increased from 18% to 44% after training. CONCLUSION: There was an overall improvement in the knowledge about dementia among the health care professionals following the training, which was reflected in the increase in referrals to the memory clinic. Although the initial results appeared to be promising, a multicentre study is warranted to conclude that the training had been effective.

Rising from the dead!

A report of a patient with Lazarus phenomenon (the return of spontaneous circulation after cardiopulmonary resuscitation) following cardiac arrest (myocardium ischemia) is presented. A 65 year patient was found unconscious at home. He taken to the emergency department On arrival he was unconscious, his pupils fixed and dilated bilaterally. Resuscitation proceeded for 55 minutes. He was then pronounced dead. Forty minutes later spontaneous breathing was noted and his blood pressure was 110/48 and heart rate 90bpm. He survived a further 13 days in the coronary care unit. The implications for management of cardiac arrest in the emergency and medical department are discussed.

Increase in tumour-infiltrating lymphocytes with regulatory T cell immunophenotypes and reduced zeta-chain expression in nasopharyngeal carcinoma patients.

The pathological significance of the mechanisms of tumour immune-evasion and/or immunosuppression, such as loss of T cell signalling and increase in regulatory T cells (T(regs)), has not been well established in the nasopharyngeal carcinoma (NPC) microenvironment. To evaluate the T(reg) immunophenotypes in tumour-infiltrating lymphocytes (TILs), we performed a double-enzymatic immunostaining for detection of forkhead box P3 (FoxP3) and other markers including CD4, CD8, and CD25 on 64 NPC and 36 non-malignant nasopharyngeal (NP) paraffin-embedded tissues. Expression of CD3 zeta and CD3 epsilon was also determined. The prevalence of CD4(+)FoxP3(+) cells in CD4(+) T cells and the ratio of FoxP3(+)/CD8(+) were increased significantly in NPC compared with those in NP tissues (P < 0.001 and P = 0.025 respectively). Moreover, the ratio of FoxP3(+)/CD25(+)FoxP3(-) in NPC was significantly lower than that in NP tissues (P = 0.005), suggesting an imbalance favouring activated phenotype of T cells in NPC. A significant negative correlation between the abundance of FoxP3(+) and CD25(+)FoxP3(-) cells (P < 0.001) was also identified. When histological types of NPC were considered, a lower ratio of FoxP3(+)/CD25(+)FoxP3(-) was found in non-keratinizing and undifferentiated carcinomas. Increased CD4(+)FoxP3(+)/CD4(+) proportion and FoxP3(+)/CD8(+) ratio were associated with keratinizing squamous cell carcinoma. A reduced expression of CD3 zeta in TILs was found in 20.6% of the NPC tissues but none of the NP tissues. These data provide evidence for the imbalances of T(reg) and effector T cell phenotypes and down-regulation of signal-transducing molecules in TILs, supporting their role in suppression of immune response and immune evasion of NPC.

Hypomutability at the polyadenine tract in SMN intron 3 shows the invariability of the a-SMN protein structure.

Recently, the axonal-SMN (a-SMN) protein, which is generated by the gene responsible for spinal muscular atrophy (SMA), SMN, has been reported. Surprisingly, the a-SMN transcript includes the entire sequence of SMN intron 3. We had expected a high frequency of insertion/deletion mutations at a polyadenine tract in this intron, since simple repetitive sequence motifs are prone to mutations. Such mutations could change the C-terminal structure of the a-SMN protein. However, our study showed that almost all individuals, including healthy individuals, SMA patients and SMA-like patients, carried only alleles with a normal polyadenine tract. Hypomutability of the polyadenine tract in SMN intron 3 suggests the existence of transcriptional mechanisms preventing alterations to the open reading frame of axonal SMN and not allowing variability in the protein structure of a-SMN.

Effect of incorporating sodium molybdate in the form of salt lick or in mixed ration on growth and performance of sheep fed palm kernel cake.

The objective of the present experiments are to determine if sheep could safely consume high amount of PKC in their diets and if sheep's consumption of PKC require a chelating agent to tie up the high copper level in PKC to protect it against toxicity. Two experiments were carried out. Experiment 1, with four treatment groups of animals, using from 30 to 100% PKC, mixed with other feed ingredients except minerals. Mineral mixtures were separately mixed with Sodium Molybdate, acting as the chelating agent, and the mineral was offered in separate feed troughs ad.libitum, in the pens for each animal group. Although all groups gave high ADG, the animals in the 100% group had high copper in their blood, which were above4 the normal physiological level at the end of the experimental duration of three months. Three animals from this group died and their liver copper contents were very high. The groups fed up to 72% PKC in their ration did not show any significant elevation of copper or toxicity. Sheep fed similar proportions of ingredients in experiment 2, but with Molybdate incorporated together into all the four similar rations as in experiment 1, did not show any signs of toxicity or elevated blood copper. The animals in all groups produced high ADG. The experiment proved that sheep can take up to 100% PKC in their diet, but a chelating agent must be incorporated into the feed to ensure its sufficient uptake to protect it against toxicity. Giving Molybdate separately from the feed would not ensure sufficient intake of the chelating agent voluntarily. This would result in copper toxicity in the animals.