1.Optimization of cell density and LPS concentration for the evaluation of nitric oxide production on BV-2 in a Griess assay
Nasim Karimi Hosseini ; Shinsmon Jose ; Sharmili Vidyadaran ; Syafinaz Amin Nordin
Malaysian Journal of Medicine and Health Sciences 2014;10(2):1-8
Introduction: Production of nitric oxide (NO) is one of the main responses elicited by a variety of
immune cells such as macrophages (e.g. microglia, resident macrophages of brain), during inflammation.
Evaluation of NO levels in the inflammatory milieu is considered important to the understanding of the
intensity of an immune response; and has been performed using different methods including the Griess
assay. To assay NO in culture, an appropriate number of cells are stimulated into an inflammatory phenotype.
Common stimuli include lipopolysaccharide (LPS), IFN-γ and TNF-α. However, overt stimulation could
cause cell cytotoxicity therefore an ideal concentration of LPS should be used. Objective: To set-up a
model of BV-2 cell activation that allows the assay of detectable levels of NO. Optimization of BV-2
microglia cell density and LPS concentrations after stimulation by bacterial lipopolysaccharide (LPS)
for the Griess assay is demonstrated in this study. Methods: BV-2 microglia were cultured at different
cell densities, and treated with LPS at three concentrations (1, 5, 10 µg/ml). NO production in culture
supernatants were then measured at 18, 24, 48 and 72 hours. Moreover, methyl tetrazolium assay (MTT)
was also performed to ensure that NO measurement is performed at no-cytotoxic concentrations of
LPS. Results and Conclusions: NO production follows a temporal pattern. The density of 25000 cells/
well was the ideal seeding density for NO evaluation in BV-2 cells. BV-2 stimulation by LPS is dose
dependent, and NO levels are increased proportional to the LPS concentration up to 1.0µg/ml, whereas
the higher LPS concentrations are associated with decreased cell viability may be caused by the high
toxic levels of LPS or NO. Although Griess assay has been commonly used by the scientists, however,
optimization of its parameters on BV-2 cells will be useful for the experiments which will be performed
on this particular cell line. The optimized pattern of Griess assay on BV-2 cells was achieved in this
study, hence easier and more practical for the future scientists to perform Griess assay on BV-2 cells.
Nitric Oxide
2.Comparative Analysis of Inflammatory Markers Produced by Macrophages Inoculated with Invasive and Colonizing Strains of Streptococcus Agalactiae (Group B Streptococcus) and Evaluation of Patients’ Clinical Data
Nassim Karimi Hosseini ; Sharmili Vidyadaran ; Shinsmon Jose ; Narges Eskandarian ; Zalina Ismail ; Syafinaz Amin Nordin
Malaysian Journal of Medicine and Health Sciences 2017;13(1):55-60
Introduction: Group B Streptococcus (GBS), infection and recurrence in newborns and pregnant women can lead to
chronic medical illness resulting in significant morbidity, and mortality. Pathogenesis of GBS may be due to reasons
such as activation of the immune system, followed by the production of inflammatory markers and toxic components
by immune cells including macrophages. Methods: The studies on invasive and colonizing GBS strains inoculated
either with peripheral or brain macrophages, the expression of nitric oxide (NO), cell viability, and CD40 were
also measured by Griess assay, methyl tetrazolium assay (MTT), and flow cytometry, respectively. Furthermore, the
clinical manifestations of the selected patients were also assessed for this study. Results: Outcome of inflammatory
markers studies, after GBS inoculation indicated that, invasive GBS strains induced higher inflammatory markers
in comparison to colonizing GBS strains. Furthermore, patients’ clinical data showed that patients with invasive
GBS infections had severe condition unlike among patients with colonizing GBS strains. The fatality rate in patients
with invasive GBS strain were 30.8% while there was no death among carriers. Conclusion: This study, aimed to
understand the immune response to GBS, and strengthen the knowledge on GBS pathogenesis. It was concluded
that invasive GBS strains not only showed higher expression of inflammatory markers on immune cells but also had
higher pathogenesis effect in comparison to colonizing GBS strains.
Streptococcus agalactiae
;
Pregnancy
3.Colistin-associated nephrotoxicity among patients in intensive care units (ICU) of hospitals in Selangor
Rashizal Sazli Mohd Rasidin ; Ami Fazlin Syed Mohamed ; Wan Mazuan Wan Mahmud ; Ling Siew Mei ; Aidalina Mahmud ; Syafinaz Amin Nordin
The Medical Journal of Malaysia 2017;72(2):100-105
Introduction: The increasing trend of extensively drugresistant
gram negative bacteria responsible for nosocomial
infections has prompted resurgence colistin usage.
Colistin-induced nephrotoxicity is a concern with disparity
in the reported rates between previous studies. This study
aims to evaluate colistin-induced nephrotoxicity among
Malaysian population.
Methods: The medical records of ICU patients receiving
colistin therapy in Hospital Serdang and Hospital Sungai
Buloh from 2010 to 2012 were retrospectively reviewed.
Demographics data, treatment characteristic as well as
culture result and creatinine level were documented.
Nephrotoxicity was determined based on RIFLE criteria.
Results: A total of 100 patients were included. Median daily
dose, cumulative dose and duration of colistin therapy were
3.0 MIU (IQR: 4, range 1-12), 17.8 MIU (IQR: 31.5, range 2-180)
and seven days (IQR: 4, range 1-30). Nephrotoxicity was
found in 23% of the study population. All cases were
reversible but marginally associated with higher mortality.
No statistical association exist between age, gender and
race as well as administration routes with nephrotoxicity by
univariable analysis. The association of dose and duration
with nephrotoxicity was also not significant by univariable
analysis. After adjustment for confounders, statistical
association between the independent variables and
dependent variable remains not significant.
Conclusion: Lower dose and shorter duration in local
settings contribute to lack of association between colistin
therapy and nephrotoxicity in this study. Higher dosing
regimen with loading dose application has been introduced
in the latest National Antibiotic Guideline. Further
evaluation of colistin-induced nephrotoxicity and potential
risk factors is therefore warranted.
Colistin
;
Intensive Care Units
4.Staphylococcus aureus Nasal Carriers Among Medical Students in A Medical School
Syafinaz Amin Nordin ; Nur Ain Nabila Za'im ; Nurul Nadzirah Sahari ; Siti Fatimah Jamaluddin ; Shahram Ahmadi ; Mohd Nasir Mohd Desa
The Medical Journal of Malaysia 2012;67(6):636-638
Staphylococcus aureus is usually considered a colonizer
but can result in infections under favourable conditions,
especially in the healthcare setting. Healthcare workers can be colonized by S. aureus, and may transmit them to
patients under their care. We conducted a cross sectional
study to determine the prevalence of S. aureus nasal
carriers among medical students in Universiti Putra
Malaysia (UPM) (from January to June 2011). Our study
involved 209 medical students comprising of 111 and 97 preclinical and clinical students respectively. A selfadministered questionnaire was distributed and nasal
swabs were collected. Upon identification, the antibiotic
susceptibility of the isolates was examined followed by
categorical analysis (Chi-square and Fisher’s exact tests)
with factors associated with S. aureus nasal carriage.
Twenty one (10%) S. aureus strains were isolated from 209
nasal swab samples. 14 isolates were from pre-clinical
students while the remaining seven were from clinical
students. There was no significant association between
gender, ethnicity, health status, skin infection and students’ exposure to hospital environment with S. aureus nasal carriage (p>0.05). Nineteen (90.5%) isolates were resistant to penicillin and there was also no significant association between penicillin resistant and the students’ groups. One (5.3%) isolate was resistant to erythromycin. There was no methicillin-resistant S. aureus isolated in this study.
5.Clinical presentation and microorganisms sensitivity profile for diabetic foot ulcers: a pilot study
Nur Hilda Hanina ABD Wahab ; Intan Nureslyna Samsudin ; Syafinaz Amin Nordin ; Zalinah Ahmad ; Lailatul Akmar Mat Noor ; Anand Sobhraj Devnani
The Medical Journal of Malaysia 2015;70(3):182-187
SUMMARY
Introduction: Patients suffering from diabetes mellitus (DM)
frequently present with infected diabetic foot ulcers (DFU).
This study was done to record the anatomical site and the
grade of ulcers according to Wagner’s classification and to
culture the microorganisms from the ulcers and determine
their antibiotic sensitivity.
Materials and methods: Prospective study was conducted
on 77 diabetic patients who were admitted with DFU from
June until December 2011. Patients with end stage renal
failure, those who had previous vascular surgery on the
involved limb, or hyperbaric oxygen or maggot therapy for
the ulcers, or had unrelated skin diseases around the
involved foot were excluded from the study. Specimens for
culture were obtained by a sterile swab stick or tissue
sample was taken from the wound with sterile surgical
instruments.
Results: Wagner’s grade III and IV ulcers were most
common. Majority of the ulcers involved toes (48%). Gram
negative microorganisms were predominantly isolated
(71.1%). Gram positive microorganisms were less frequently
cultured (27.7%). Fungus was cultured from one sample
(1.2%). Gram negative microorganisms were sensitive to
aminoglycosides, cephalosporins or β-lactamase inhibitors.
More than 40% were resistant to ampicillin. Gram positive
microorganisms were sensitive to cloxacillin. MRSA were
sensitive to vancomycin.
Conclusion: Empirical use of antibiotics should be curtailed
to prevent development of drug resistant strains of
microorganisms and MRSA. We suggest use of antiseptic
solutions to clean the ulcers until antibiotic sensitivity
report is available. Results of our altered treatment regimen
we plan to publish in a later study.
Diabetic Foot
;
Foot Ulcer
6.Multilocus Sequence Typing Analysis of Invasive and Non-Invasive Group B Streptococcus of Hospital Origin in Malaysia
Menagah Ezhumalai ; AbdulRahman Muthanna ; Zarizal Suhail ; Nurul Diana Dzaraly ; Syafinaz Amin-Nordin ; Mohammad Noor Azmai Amal ; Mohd Nasir Mohd Desa
Malaysian Journal of Medical Sciences 2020;27(1):134-138
The aim of this study was to study the genotype of a hospital collection of Group B
Streptococcus (GBS) from invasive and non-invasive sites. Fifty-one pre-characterised human
of GBS were re-identified and further analysed by multilocus sequence typing (MLST) in relation
to previously published serotypes. Fifteen sequence types (ST) were found with ST1 being the
most predominant. ST1 was also associated with majority of the invasive isolates. The genotypic
distribution patterns of GBS in this study were largely in agreement with previous reports from
other countries indicating the tendency of certain genotypes to prevail in human infection settings.
7.An update on Gardneralla vaginalis associated bacterial vaginosis in Malaysia
Younus Khairi NADA ; Gopinath RENU ; Jegasothy RAVINDRAN ; Nordin Amin SYAFINAZ ; Belkum van ALEX ; Mary NARCISSE ; Neela Kumari VASANTHA
Asian Pacific Journal of Tropical Biomedicine 2017;7(9):831-835
Objeetive:To update the status of Gardnerella vaginalis (G.vaginalis) as a causative agent of bacterial vaginosis (BV) in Malaysia and to define its epidemiology,metronidazole resistance and virulence properties.Methods:It is a single-centre (Gynaecology clinic at the Hospital Kuala Lumpur,Malaysia) prospective study with laboratory-based microbiological follow up and analyses.Vaginal swabs collected from the patients suspected for BV were subjected to clinical BV diagnosis,isolation and identification of G.vaginalis,metronidazole susceptibility testing,vaginolysin and sialidase gene PCR,Piot's biotyping and amplified ribosomal DNA restriction analysis genotyping.Results:Among the 207 patients suspected for BV,G.vaginalis was isolated from 47 subjects.G.vaginalis coexisted with Trichomonas vaginalis and Candida albicans in 26 samples.Three G.vaginalis isolates were resistant to metronidazole.Biotyping revealed 1 and 7 as the common types.Amplified ribosomal DNA restriction analysis genotype Ⅱ was found to be more common (n =22;46%) than Ⅰ (n =12;25.53%) and Ⅲ (n =13;27.6%).All genotype Ⅰ and Ⅲ isolates carried the sialidase gene,while 91.6% and 84.6% contained the vaginolysin gene.Genotype Ⅰ was significantly associated with postgynaecological surgical complications and abortions (P =0.002).Conclusions:The existence of pathogenic G.vaginalis clones in Malaysia including drug resistant strains should not be taken lightly and needs to be monitored as these may bring more complications especially among women of child bearing age and pregnant women.
8.Genotypic and phenotypic characterization of methicillin resistance determinants and β-lactamase in Staphylococcus species
Abdul Rahim Abdul Rachman ; Norhidayah Mat Azis ; Pung Hui Ping ; Zarizal Suhaili ; Syafinaz Amin Nordin ; Zulkefley Othman ; Mohd Nasir Mohd Desa
Malaysian Journal of Microbiology 2017;13(4):308-317
Aims: To characterize the genotypic distribution of mec complex, bla complex, methicillin-resistance level (cefoxitinMIC)
and β-lactamase activity in carriage methicillin-resistant Staphylococcus species for a potential correlation.
Methodology and results: Biochemical test, 30 µg cefoxitin diffusion disc test, cefoxitin E-test, mec and bla complexes
distributions, Pbp2a and β-lactamase assays were conducted to characterize phenotypic and genotypic of MRSA and
MRCoNS in our collection. Phylogenetic tree was constructed using MEGA6 software to trace the diversity of blaZ gene
of MRSA and MRCoNS. Sixteen MRSA and nineteen MRCoNS were identified by biochemical tests followed by 30 µg
cefoxitin antibiotic disc susceptibility test and mecA gene screening. Twenty nine isolates carry complete mecA genes
(2.1 kb), incomplete mec regulator (negative or truncated) and positive Pbp2a assay for both MRSA and MRCoNS. Only
MRCoNS SC177 isolate with cefoxitin MIC of 32 µg/mL carries complete mec complex. Thirty-one of thirty-five isolates
carry complete bla complex (blaZ, blaRI, blaI) with 10 MRSA produce strong β-lactamase and cefoxitin MIC of ≥12
µg/mL. Only 4 MRCoNS with cefoxitin MIC of ≤8 µg/mL produce strong β-lactamase. The diversity of blaZ gene was
demonstrated by phylogenetic analysis and unusual amino acid mutation at position 145 for MRSA SA60 isolate may
compromise its β-lactamase activity with low cefoxitin MIC level (2 µg/mL).
Conclusions, significance and impact of the study: Isolates that carry complete complete mecA gene were largely
consistent with the expression of Pbp2a. Nevertheless, there is no clear correlation of mec regulator genes in relation to
cefoxitin-MIC in both methicillin resistant (MR) Isolates that carry Staphylococcus species. On the other hand, various
expression level of β-lactamase may correlate with cefoxitin-MIC level in MRSA as compared to MRCoNS.
9.Approaches of Learning among Medical Undergraduates of Faculty of Medicine and Health Sciences, Universiti Putra Malaysia in 2016
Nurul Mursyidah ZAKARIA ; Nur Asyiqin Jasman Farok ; Siti Khadijah ADAM ; Syafinaz Amin NORDIN
Malaysian Journal of Medicine and Health Sciences 2018;14(1):1-6
Introduction: Challenge arises for medical undergraduates as the subject of medicine is intricate and extensive. Although students come from the same pool of excellent academic background, the medical undergraduates are still prone to failure in exams, resulting in them repeating the year of study or even having the thought of changing to other courses. In order to cope with the programme, students may adopt learning approaches that would help them to go through the programme.Therefore, it would be interesting to explore the learning approaches of medical students in Universiti Putra Malaysia. Methods: The purposes of this study were to determine the learning approach of medical undergraduates of Faculty of Medicine and Health Sciences, Universiti Putra Malaysia (FMHS, UPM) together with its associated factors, which were socio-demographic characteristics and learning environment. This research was a cross-sectional study where the sample size calculated was 554. Self-administered questionnaires were given to the respondents chosen by simple random sampling. The socio-demographic characteristics were analysed using descriptive statistics such as frequency and percentage. Chi-square test was used to analyse the association between the variables. Results: The response rate was 83% (460 respondents agreed to participate). The majority of the respondents were females (73.7%), Malay (67%), and in their clinical years of study (58.7%). Overall, most students preferred deep approach (DA) of learning (49.6%), followed by strategic approach (29.1%) and surface apathetic approach (21.3%) of learning. There were statistically significant associations between learning approach and gender (p=0.005), as well as between learning approach and year of study (p=0.037). Conclusion:Our study showed an association between learning approaches and year of study and gender. DA of learning was the preferred learning approach in medical students at FMHS, UPM. This approach of learning, where students learn to understand the subject matter, may result in students become effective learners. Their understanding about the subject matter will be applicable to their clinical practice in the future.
10.It was a Fungal Culprit all Along - A Case Report
Navin Kumar Devaraj ; Syafinaz Amin Nordin
Malaysian Journal of Medicine and Health Sciences 2020;16(SP 1, September):35-37
Sporotrichosis is a relatively uncommon fungal infection in human as compared to other fungal infectiction such as tinea corporis or cruris. The fungus can be found in the environment and on plant matter. Sporotrichosis
usually results from trauma to the skin followed by exposure to the fungus. It can also be spread by cat scratch or bite.
Sporotrichosis usually presents as multiple skin nodules arranged in a linear fashion over the limbs, often with
associated lymphadenopathy. This calls for importance of full physical examination and a high index of suspicion in order to diagnose this infection that can either involve the skin or the lymphatic channel or both. If misdiagnosed, treatment can be delayed and lead to prolonged suffering of the patient. This is an interesting case report about a 43-year-old woman who presented with left axillary swelling and decreased range of movement of the corresponding shoulder which was feared initially to be linked to breast cancer. However, good clinal acumen lead to the actual diagnosis of sporotrichosis.