Complete blood count (CBC) is used broadly to screen individual's general health status. Some inherited red blood cell (RBC) disorders influence the RBC parameters. Mean corpuscular volume (MCV) and mean corpuscular haemoglobin (MCH) are amongst the important RBC parameters used in thalassaemia-haemoglobinopathy screening [1-2]. Globin chain disorders and Southeast Asian Ovalocytosis (SAO) are common RBC disorders in Southeast Asian countries [3]. We evaluated the RBC parameters in patients with Hb E and those with SAO co-inheritance.
A total of 33 from 1500 Malay patient’s samples that were sent for thalassaemia-haemoglobinopathies screening in Hospital Kuala Lumpur (HKL) were identified and consented (30 cases with Hb E and 3 cases with co-inheritance of Hb E and SAO). The inclusion criteria were Malay patients with MCV and MCH levels less than 78 fL and 27 pg respectively with presence of oval and stomatocytic RBCs in the peripheral blood film. DNA extraction was performed in samples suspected of having co-inheritance of SAO and Hb E. Primers 198 and 199 (AIT biotech Pte Ltd. Singapore) were designed for SAO detection [4], [5]. Hb E mutation was detected using ARMS PCR [6].
SAO was characterised by presence of an in frame 27bp deletion in exon 11 of the band 3 gene. A band of 175bp was observed in normal subjects and two bands, 175bp and 148bp were observed in heterozygous SAO subjects (Fig. 1).
The aim of the study is to evaluate the effects of contact and non-contact laser photocoagulation (LP) on ocular surface changes and Ocular Surface Disease Index (OSDI) score in patients with proliferative diabetic retinopathy (PDR). This was a single center, prospective, randomised, parallel-controlled trial of pilot study in Hospital Universiti Sains Malaysia between June 2013 and May 2014. Eye with PDR was selected and randomised into 2 groups (Contact LP group and Non-contact LP group) by using random sampling envelope method. Contact LP group was treated with contact LP via slit lamp laser delivery system. Non-contact LP group was treated with non-contact LP via binocular laser indirect ophthalmoscopy system. Main outcome measures were Schirmer test value, tear film break-up time (TBUT) and OSDI score at baseline and at 3 months post laser therapy. Statistical analyses were performed using SPSS version 22.0. A total of 60 eyes were recruited (30 eyes in Contact LP and 30 eyes in Non-contact LP). Contact LP showed significant reduction of TBUT (p = 0.038) and significant increase in mean OSDI score (p = 0.001) at 3 months post laser therapy. However, there was no significant difference of mean change of Schirmer test value and TBUT between the two groups except for OSDI score (p = 0.044). Both mode of laser deliveries (contact LP and non-contact LP) showed comparable effects on ocular surface disease in PDR patient that underwent laser pan retinal photocoagulation.
The incidence of HbE/beta (HbE/β) thalassaemia is increasing in Asian countries, including Malaysia. HbE/β thalassaemia is widely acknowledged to have a diverse phenotypic spectrum despite having the same primary genetic background [1,2,3]. Thus, there are HbE/β thalassaemia patients who receive unnecessary treatments which leads to side effects [4], reduced quality of life and wasting health care resources. Ideally, the treatment and management of thalassaemia patients are individually tailored in order to minimise side effects and optimise health care costs. Genetic variants have been widely acknowledged to influence the variability of human phenotypes. Presence of unique genetic modifiers are believed to cause the diversity in HbE/β thalassaemia severity. Milder disease course has been found to be highly associated with Xmn1-Gγ polymorphism (rs7482144), a SNP at HBG2 promoter [1,5,6,7]. So far, there is no association study between Xmn1-Gγ polymorphism and HbE/beta thalassaemia disease severity in Malaysia. This study aims to optimise PCR-RFLP technique for detection of Xmn1-Gγ polymorphism, to determine the frequency of Xmn1-Gγ polymorphism in HbE/β thalassaemia patients and finding its association with the severity of HbE/β thalassaemia patients. This hospital-based cross-sectional study was performed using archived genomic DNAs from 58 subjects with their respective research pro formas. Selected datas were extracted from the pro formas in order to classify patients into 3 disease severity groups using the scoring system by Sripichai et al., (2008) based on 6 parameters. The archived genomic DNAs were genotyped employing Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) technique. The genotypes were categorised into homozygous variant, heterozygous and homozygous wild type. The genotypes detected were then validated using DNA sequencing analysis. Appropriate statistical analysis was used to determine the association of Xmn1-Gγ polymorphism with the clinical severity of HbE/β thalassaemia. This study had successfully optimised the PCR-RFLP technique for detection of Xmn1-Gγ polymorphism. Out of 58 subjects, the Xmn1-Gγ polymorphisms were detected in 40 subjects (69%) with the majority being heterozygous (CT) (n=38, 66%) and there were only 2 (3%) homozygous variant (TT) subjects. Homozygous wild type (CC) were detected in 18 (31%) subjects. There were no significant association of Xmn1-Gγ polymorphism with the severity of HbE/β thalassaemia patients with p-value of 0.65 for genotype and 0.58 for allele, respectively. In conclusion, this study showed no significant association of Xmn1-Gγ polymorphism with milder disease severity of HbE/β thalassaemia patients. This can be a true finding for the patients in North East Malaysia or due to small sample size. Thus we recommend to have a larger study in order to validate the association of Xmn1-Gγ polymorphism with HbE/β thalassaemia severity. In addition, there may be other genetic factors that interact with Xmn1-Gγ polymorphism as it was not possible to consistently predict phenotype and severity from the presence of Xmn1-Gγ polymorphism alone.
Odontogenesis is a complex process regulated by both genetic and molecular controls. The development of a tooth in the embryo stage is controlled by a series of signals which occur between tooth-forming epithelium and neural crest-derived ectomesenchyme. Though many genes are involved in tooth formation involving major signalling molecules, the bone morphogenetic protein and fibroblast growth factor are the most important ones involved in odontogenesis. Supernumerary tooth occurs because of imbalance in the expression of the signalling pathways and their inhibitors. This review highlights the various signalling molecules that play a role in odontogenesis in order to provide a better understanding on of the molecular mechanisms involved in the formation of supernumerary tooth in humans.
Muscular dystrophy is a group of diseases that result in progressive muscle weakness and atrophy. Duchenne Muscular Dystrophy (DMD) is classified as dystrophinopathy and is an X-linked recessive disease. It is caused by alterations in the dystrophin gene at Xp21.2 encoding 79 exons [1]. It is characterised by progressive muscle wasting that begins at 3 to 5 years, delay in motor development and eventually wheelchair confinement followed by premature death at about 30 years from cardiac or respiratory complications [2]. Genetic etiology of cases of DMD in Malaysia are still scarcely reported. Here, we report the genetic cause in the case of an 11-year-old Kelantanese Malay boy who has progressive muscle weakness since 5 years old. He has difficulty in getting up from sitting and supine position also in climbing up stairs until 1st floor. He has a strong family history of DMD and musculoskeletal problems. His younger brother was diagnosed with DMD by molecular analysis and his maternal uncle died at the age of 16 with musculoskeletal problems but was never investigated. Physical examination revealed no dysmorphic features, positive Gower sign with absent tounge fasciculation. On neurological examination, tendon reflexes and muscle tone for limbs were normal. Muscle power for bilateral upper limbs were normal, however, bilateral lower limbs showed slight reduction in muscle power with calf hypertrophy.
An 11- month-old girl with accidental findings of pale and hepatosplenomegaly. She was the last child of three siblings from a non-consanguineous marriage. The father and the mother were Hb E trait and Hb Constant Spring (Hb CS) trait respectively. Clinically the child was small for age with frontal bossing and hepatosplenomegaly. Sytemic examination was unremarkable. Her full blood picture showed moderate hypochromic microcytic anaemia with marked anisopoikilocytosis (Hb of 7.1g/dl, MCV of 44.6 fl, with MCH of 13.8 pg and RDW-CV of 24.0%). Quantitation of haemoglobin by using High Performance Liquid Chromatography (HPLC) and gel electrophoresis report showed that the patient has compound heterozygous E/ß+ thalassaemia with Hb H-CS. She had increased of Hb A2/E (28.9%), and Hb F (11.2%) with presence of pre-run peak and a tiny peak at C window. Gel Electrophoresis by using agarose gel at alkaline pH discovered prominent A2 band and fast band to the left of Hb A band. H inclusions were positive. Further confirmation of diagnosis was done by molecular study. Alpha molecular study using Multiplex GAP PCR showed heterozygous --/SEA deletion (Fig. 1), while beta molecular study using Multiplex Amplification Refractory Mutation Systems (ARMS) revealed Cd 26 (G-A) and CAP +1 (A-C) mutations [Fig. 2]
Hemoglobin (Hb) E is common in Southeast Asia [1]. HbE disorders may be found heterozygous (AE) which usually asymptomatic, homozygous (EE) and compound heterozygous state with widely variable clinical features, ranging from transfusion dependence to a complete absence of symptoms [2]. Considering her history, clinical findings and investigations, the most likely diagnosis in our case is Compound heterozygous E/ß+ thalassaemia with Hb H-CS. She had moderate hypochromic microcytic anaemia, raised Hb A2/E and Hb F with presence of pre-run peak and a tiny peak at C window support the diagnosis. Unfortunately, we’re unable to confirm the presence of Hb CS in view of no modalities available in our setting. However, with the family history of mother with Hb CS trait, the presence of Hb CS in this patient cannot be denied as a factor contributing to Hb H disease. Previous study reported Hemoglobin Constant Spring is often missed by routine laboratory testing, especially in subjects with co-inheritance of β-thalassaemia or β-variants. Hb CS detection clearly seen in capillary electrophoresis compared to HPLC [3]. As in this case only a very tiny peak of Hb CS noted on the HPLC. The molecular analysis for detection of Hb CS should be performed as for confirmation test. Hb H-CS has a severe phenotype than a deletional Hb H disease [4]. The diagnosis was confirmed by molecular analysis. Hence, genetic testing and family study are of particular importance to establish the exact genetic defect causing the abnormal Hb in this patient.
In view of thalassaemia is common in our region, it is important to identify complete genotyping to provide proper management, make clinical predictions and improve genetic counseling.
Gene expression is the most fundamental level at which the
genotype gives rise to phenotype. The development of human salivary
exosomes has become one of the promising researches to improve cell-based
tissue engineering but their functions in human periodontal ligament fibroblast
(HPdLF) cells are not well studied. To study the effect of human salivary
derived exosomes on the gene expression of HPdLF cells. In vitro, HPdLF
cells were cultured for 24 hours with 10 µg/ml of human salivary exosomes.
Determination of gene expression levels of basic fibroblast growth factor
(bFGF) and collagen type I (COL1) in the presence and absence of human
salivary exosomes in HPdLF culture was performed using quantitative reverse
transcriptase polymerase chain reaction (RT-qPCR). Human salivary
exosomes significantly upregulated bFGF gene expression but not COL1
gene in HPdLF cells after 24 hours of culture. Human salivary exosomes are
able to upregulate bFGF gene in HPdLF cells. Thus, they might have potential
to be used as an alternative biomaterial in tissue engineering for periodontal
regeneration.
Abstract—The functions displayed by exosomes derived from saliva and
other body fluids have been established. This paper studied the stability of
human salivary exosome beginning from the collection mode, storage, and its
preservation methods. Unstimulated saliva samples were collected from
healthy subjects. Protease inhibitor was added into each samples and stored
under different temperatures and at varying periods of time. The exosomes
were isolated by ultracentrifugation and confirmed by using Western Blot.
Exosome morphology was inspected by Scanning Electron Microscope (SEM)
and the protein concentration was determined using the Protein (Bradford)
Assay. The exosome particle size distribution and concentration were
calculated using Nanoparticle Tracking Analysis (NTA). The protein assay
showed no significant differences in the exosome protein concentration values
for all conditions. Western Blot analysis also showed no differences in the
presence of exosome and all the samples were positive for protein CD63.
SEM analysis showed the fine shape of exosome which is round, in vesicle
form with the size ranging between 10 nm and 100 nm. NTA determined the
individual mean and the clumping exosome size was 203 nm. Human salivary
exosomes remained intact in the absence of protease inhibitor and in different
storage temperatures.
The aims of the study were to investigate the anti-cancer effects of 5-
Aza and TSA in two leukemic cell lines (CCRF-CEM and HL-60). Inhibition
concentration of 5-Aza and TSA were measured using trypan blue exclusion
assay. 5-Aza and TSA at IC50 were treated to both CCRF-CEM and HL-60 cell
lines for 4-6 days. To confirm the inhibition effects of these agents, Annexin-V
stained cells were analyzed using flow cytometry to evaluate the apoptotic
induction. The IC50 values of CCRF-CEM were 2.01±0.1µM and 2.65±0.3µM for
5-Aza- and TSA-treated, respectively. Whereas, the IC50 values of HL-60 were
1.98±0.2µM and 2.35±0.2µM for 5-Aza- and TSA-treated, respectively. To
further substantiate the findings, the time-dependent exposure of both drugs was
studied. CCRF-CEM cells were reduced to 49.4%±5.0, 49.4%±2.5 and
41.5%±5.6 by 5-Aza; 56.5%±7.0, 45.3%±4.2 and 40.2%±4.2 by TSA treatment
at first, third and sixth day. HL-60 cells were reduced to 72.0%±4.5, 51.0%±1.5
and 40.6%±2.6 by 5-Aza at first, third and sixth day. Meanwhile, HL-60 cells
reduced to 55.6%±4.5, 45.2%±4.0 and 36.3%±2.9 by TSA at first, second and
fourth day. Both cell lines were significantly inhibited (p
The aim of this study is to determine the risk factors for retinopathy of prematurity (ROP), and also to screen Norrie Disease Pseudoglioma (NDP) gene mutation in order to determine if mutation in the NDP gene may play a role in the development of ROP among Malay premature infants. This was a case control studyamong Malay premature infants from Hospital Universiti Sains Malaysia (USM) conducted from August 2011 to May 2013. Written consent were taken from their parents before conducting the study. The stage of ROP, systemic risk factors (gestational age and birth weight) and enviromental risk factors (oxygen exposure and duration of ventilation)were reviewed from patients’medical records. DNA was extracted from venous blood and subjected to polymerase chain reaction (PCR) before direct sequencing of NDP gene. A total of 56 Malay premature infants (Case group = 28 ROP premature infants, Controlgroup = 28 non-ROP premature infants)from Hospital USMwere enrolled in this study. Out of 28 premature infants with ROP, 11 (39.3%) premature infants were in stage 3. Only 1 (3.6%) premature infant in stage 4 and 2 (7.2%) premature infants in stage 5. The gestational age (p = 0.010) and birth weight (p = 0.010) were the significant risk factors for ROP. There was no significant difference ofenvironmental risk factors between the two groups. The NDPgene mutation was not detected in Malay premature infants with ROP and also in control group. The gestational age and birth weight were important risk factors of ROP.Although NDPgene mutations were being linked to ROP but NDPgene mutation was not detected in premature infants with ROPas well as premature infants with non-ROP among Malay ethnic background.
Thalassaemia screening programme was conducted to reduce the burden of the disease [1]. Here, we describe one unexpected discovery in a 33-year-old gentleman and also the importance of DNA analysis in detecting the globin gene mutation.
Xenobiotic substance released in the environment is a concern among the public at large. The example of this xenobiotic release into the environment is xenoestrogens. Xenoestrogens have the capability to bind to the estrogen receptors in the body even at low affinity. Food, pesticides and contraceptive pills are known sources of xenoestrogens. In this study, acute toxicity test was conducted to evaluate toxicity of synthetic estrogen such as estradiol to the embryo-larvae of zebrafish model. Morphological changes in the embryo-larvae of zebrafish were also observed. The parameters that were evaluated in acute toxicity study were half lethal concentration (LC50) and few apical endpoints such as coagulation of embryos, development of pericardial edema, and eyes size. Toxicity effect of the compound was evaluated in term of behavior activity of the larvae. Results showed that certain concentration of estradiol caused toxic effects to the embryo-larvae of the zebrafish (p
44-year-old Malay lady presented with drooping of the right eyelid and worsening of left eye vision for one week duration. There was associated headache, periorbital discomfort and diplopia on left gaze. She previously had a history of recurrent optic neuritis affecting both eyes over a period of 12 years. On examination, there was right-sided partial ptosis and left exotropia. The adduction, abduction, elevation and depression of the right eye was limited. Left eye extraocular movements were full. The right eye visual acuity was 6/9, while the left eye visual acuity was perception to light, with a positive relative afferent papillary defect and a pale optic disc. The right optic disc was normal. There was reduced sensation in the trigeminal nerve distribution over the right side of the face. Neurological examination was otherwise normal. Magnetic resonance imaging of the brain and orbit revealed meningeal thickening with involvement of the right orbital apex and cavernous sinus. Blood investigations for infectious and autoimmune causes were unremarkable. She was diagnosed to have idiopathic hypertrophic cranial pachymeningitis and treated with systemic corticosteroids. The right eye extraocular motility improved, while the left eye visual acuity improved to counting finger. This case demonstrates that idiopathic hypertrophic cranial pachymeningitis may present as recurrent optic neuritis in the early phase, before radiological evidence of the disease is present. A high index of suspicion for the underlying cause is essential to prevent irreversible optic nerve damage due to recurrent optic neuritis.
The aim of this study was to design and systematically optimize
triclosan loaded nanoparticles (TCS-loaded NPs) formulation for the treatment
of periodontal disease. Triclosan (TCS) is a broad spectrum antimicrobial
agent that has been used in the treatment of the disease. The free drug has
poor aqueous solubility and therefore may encounter permeability problems
when applied to the oral cavity. Resolution IV model of Design-Expert®
software (version 10) was used for the design of experiment and optimization
of TCS-loaded NPs. The nanoparticles (NPs) were prepared using the solvent
displacement method. Effect of factors that were investigated include drugpolymer ratio, surfactant concentration, stirring speed, stirring duration, and
drug-polymer injection rate. Particle size, zeta potential, polydispersity index
(PDI) and entrapment efficiency (EE) were the critical quality attributes
selected for the study. Desirability function determined by the software for
optimized TCS-loaded NPs was 0.704. The observed particle size, PDI, zeta
potential and EE of the optimized TCS-loaded NPs was found to be 135 ± 2.3
nm, 0.1 ± 0.012, -30 ± -4 mV and 75 ± 6%, respectively. It was found that
particle size increases by elevating the concentration of polymer and
decreases with an increase in surfactant concentration and stirring speed.
Zeta potential was found to increase when surfactant concentration was
reduced. Both surfactant concentration and drug to polymer ratio were found
to negatively affect PDI while % EE was positively influenced by the increase
in polymer concentration and decrease in surfactant concentration. The use of
Design-Expert®
software helped in identifying suitable levels of critical quality
parameters for preparing improved NPs formulation for delivery of TCS into
the periodontal pocket.
Blood donation in Malaysia is practised as voluntary non-remunerated. However, recruiting and retaining blood donors remain a challenge in the transfusion service. The main aim of this study was to understand the factors affecting the return of first-time blood donors. This was a retrospective study involving 480 first-time temporarily deferred whole blood donors from National Blood Centre (NBC), Kuala Lumpur. Data of donors who were deferred from 2010 to 2014 were extracted from the Blood Bank Information System. Deferred blood donors were categorised into two main groups, namely, a group of donors who returned for blood donation and a group that did not return for the donation. Each blood donor was contacted personally via telephone. Donors who returned were younger (p < 0.001), with females in a higher proportion (61.3%) compared to males (38.8%) (p < 0.001). Singles (68.3%) were more likely to return for donation compared to married donors (31.7%) (p < 0.001). Donors who lived in urban areas were more likely to return for donation compared to donors who lived in rural areas (34.6%) (p < 0.005). The most common factor that had motivated these donors to return was self-satisfaction (29.9%), while the most common factor that hindered them from returning for donation was the lack of time (28.50%). As a conclusion, more awareness and education regarding regular blood donation should be considered to donors from a rural areas. Additionally, mobile blood donation drives should be made easier for blood donors who have a busy lifestyle.
To report on a rare case of an intralenticular foreign body which
demonstrates that use of a spring-powered airsoft gun can result in a severe
ocular injury. A 2-year-old male presented following a trauma to the left eye.
The trauma was caused by a wooden matchstick from a spring-powered airsoft
gun being shot into his eye. On examination, there was a shallow anterior
chamber with a full thickness corneal laceration, with fragmented matchstick
pieces embedded in the cornea and in the lens. The corneal foreign bodies
were removed, corneal laceration wound sutured and lensectomy done. There
were no post-operative complications, and the cornea wound healed with a
scar. A few months later, he underwent a successful secondary intraocular lens
implantation surgery with an iris claw lens. Airsoft guns are easily available to
children who are unaware of its dangers. It can cause significant ocular
morbidity despite successful surgical treatment of the injury.
Breast cancer in young pregnancy is a breast cancer diagnosed in a young lady at the age of less than 30 years old during pregnancy. Diagnosis and treatment of breast cancer during pregnancy are challenging as both maternal and fetal outcome must be considered in terms of their short and long-term effects. It requires multidisciplinary treatment. Instead, pregnancy should be preserved whenever possible, while treating the underlying breast cancer. It is important not to compromise the treatment of breast cancer because of the pregnancy. The overall therapeutic concept basically depends on timing of delivery, staging of the underlying disease, treatment mode and the impacts of treatment recommended as well as foetal outcome in relation to treatment that administered to the mother during pregnancy. We present a case of young pregnant lady at the age of 23 with breast cancer as it is a rare event and the challenges encountered in managing her and to highlight the treatment options for the patient and the baby.
Shift in the dispersion of breast cancer population continues to be a major health concern for women in the developing countries. However, prevalence and associated factors of physical activity among this population have not been explored in Malaysia. This study determined the profile and factors associated with physical activity in the breast cancer women. A multi-center cross-sectional survey was conducted among 399 breast cancer women. Socio-demographic questionnaire was used to generate socio-demographic and clinical data. Physical activity was assessed using the international physical activity questionnaire (IPAQ-SF). Sociodemographic variables were summarized by descriptive statistics. Factors associated with physical activity were identified using Logistic regression analyses. Majority of the participants were married (83.9%), had surgery (92.4%) and menopause (74.1%). The overall physical activity prevalence was 39.4%. Non formal education (Adjusted Odds Ratio [AOR] 5.80; 95%CI: 1.65, 52.15;p=0.017), having had surgery (AOR 0.18; 95%CI: 0.04, 0.79; p=0.023),
To report a rare case of an elderly gentleman who presented with herpes zoster ophthalmicus, complicated with persistent hyphema and orbital apex syndrome. A 75-year-old Malay gentleman presented with left herpes zoster ophthalmicus that was complicated with complete ophthalmoplegia and ptosis. He developed total hyphema in the affected eye with a secondary elevated intraocular pressure after a week. He was treated with oral acyclovir and topical corticosteroids. However, the total hyphema persisted that required an anterior chamber washout surgery. Herpes Zoster Ophthalmicus complicated with persistent hyphema and orbital apex syndrome is rare and very challenging to manage. Radiological imaging is important to exclude other causes of OAS. It is recommended to treat HZO with systemic acyclovir for a longer duration in view of ocular and neurological involvement.
In Malaysia, Sabah population constitutes the most number of β-thalassaemia cases ranging from asymptomatic to transfusion dependent. Filipino β°-deletion has been reported as the predominant mutation in Sabah [1]. Despite having the same primary mutation, co-inheritance of genetic variants at HbF quantitative trait loci of HBS1L-MYB intergenic region may cause variability in clinical features by affecting the haemoglobin (Hb) subtypes level, especially HbF. Study suggested that MYB would activate γ-globin repressor gene directly and subsequently initiate the molecular HbF repression mechanisms. Polymorphisms within HBS1L-MYB intergenic region would inhibit binding of transcription factor on MYB and leading to elevation of HbF levels [2]. This can act as an ameliorating factor in the clinical presentation of β-thalassaemia patients [3]. This study aimed to elucidate the association of Hb subtypes levels with three HBS1L-MYB variants among 134 Filipino β°-deletion carriers. PCR-RFLP analysis was done for HBSIL-MYB rs4895441 (A→G) while tetra-primers ARMS PCR analysis was done for HBSIL-MYB rs9399137 (T→C) and rs11759553 (A→T) (Fig.1).