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Articles by S. Sahidan
Total Records ( 5 ) for S. Sahidan
  Z.A. Shahrul Hisham , S. Sahidan , M.A.W. Rohaya , M.Y. Siti Afeefah , Z.A. Intan Zarina , J.A.N. Nor Hidayah , R.M.A. Nadiah and Z.A. Zaidah
  Problem statement: DNA samples from fourteen modern human bloods (seven males and seven females) and two ancient skeletal samples excavated from Kalumpang Island and Dungun, Peninsular Malaysia were subjected for molecular genders determination using specific primers of human AMELX and AMELY. Approach: A standard multiple PCR mixture with forward primer and either a human X-specific reverse primer for AMELX or a human Y-specific reverse primer for AMELY amplifications were used to assess the presence of these genes in male and female samples. Results: PCR amplification of a modern male sample yielded 329 and 235 bp bands, whereas a modern female sample only 329 bp band. The Kalumpang Island sample produced two positive bands of AMELY and AMELX. After reamplification of the Dungun sample, only an AMELY band was visible. All amplified bands were cloned into TA plasmid and sequenced. BLAST analysis showed that the 329 bp band is AMELX, while the 235 bp product is AMELY. Conclusion/Recommendations: The skeletal remains of both Kalumpang Island and Dungun samples from west and east of Peninsular Malaysia respectively, are males.
  A. Nurul Atikah , Z. Zulkiflie , M.A.W. Rohaya , S. Sahidan , Z.A. Intan Zarina , M.Y. Nurul Yuziana , M. Zulkifli and Z.A. Shahrul Hisham
  Tranfection is a method that randomly introduce foreign DNA into cells either to produce genetically modified cells or to produce recombinant protein. However, efficient transfection is needed for expression of recombinant protein in mammalian cell. This study aim is to identify the transfection efficiency of novel expression vectors by qualitative and quantitative analyses. The novel expression vectors were named, pZAAGFP (without interested element), pZAAH1C (with integrated element) and pZAAM956 (with enhancer element). Chinese Hamster Ovary (CHO) cells were transfected with 3 different novel expression vectors by using Lipofectamine LTX. After 24 h, these cells were observed under confocal fluorescence microscope. The frequency of both transfected and non-tranfected cells were determined. Then, the intensity of Green Fluorescent Protein (GFP) was quantified using fluorescence reader at emission wavelength 506 nm and excitation wavelength 500 nm. Qualitative analysis showed that all novel expression vectors were able to express GFP. More than 80% CHO cells had been successfully transfected with all of the novel expression vectors stated. Quantitative analysis also showed that GFP intensity of both novel expression vectors named, pZAAH1C and pZAAM956 were higher than commercial vector and the GFP intensity of pZAAGFP was about the same as commercial vector. This finding indicates that all novel expression vectors are able to express the GFP gene and both qualitative and quantitative analyses can be applied to determine transfection efficiency.
  Z.A. Intan Zarina , Z.A. Shahrul Hisham , M.A.W. Rohaya , S. Sahidan and Z.A. Zaidah
  The aim of this study is to differentiate peripheral blood mononucleated cells that have been isolated from ICR mice`s (Mus musculus)peripheral blood into mature osteoblast and osteoclast cells with addition of growth factors. The mononucleated cells were isolated by density centrifugation using Ficoll-PaqueTM Plus. The culture mediums were then supplemented with growth factors; 50 ng mL-1 RANKL and 25 ng mL-1 M-CSF to differentiate into osteoclast cells. On the other hand, osteoblast assay, 50μg mL-1 ascorbic acid and 10 mM β-glycerophosphate were added to support differentiation. For control, the same cells were used without supplementation of respective growth factors. Biochemical analysis for osteoblast, i.e., Alkaline Phosphatase (ALP) activity was determined on day 3-14. The results showed that the ALP activity in the differentiation medium is significantly increased (p<0.05) on day 10 and day 14 as compared to control, i.e., cells without growth factors. Tartrate Resistant Acid Phosphatase (TRAP) activity that represented as osteoclast biomarker also showed significant increased (p<0.05) from day 3 until day 10 in the present of RANKL and M-CSF. The viability of differentiated cells also showed that the cells were able to survive until 10 to 14 days in the presence of respective growth factors without significant increased in respective differentiation medium. RT-PCR analysis on isolated RNA from mononucleated cells after 14 and 10 days in their differentiation medium showed that the osteoblast and osteoclast were expressing ALP (~373 bp) and TRAP (~281 bp) gene, respectively. Mononucleated cells originated from peripheral blood have the potential to differentiate into osteoblast and osteoclast cells in the presence of specific growth factors. The respective cells are primitive enough to differentiate into two distinct types of mature cells hence can be categorized as multipotent stem cells.
  M.A.W. Rohaya , S. Sahidan , Z.A. Zaidah , H. Fahrul Zaman , A.W. Nuraliza and Z.A. Shahrul Hisham
  This study aims to determine the best storage condition for salivary lactate dehydrogenase due to decreased in native enzyme’s activity after seven days of storage. Saliva samples were collected from five healthy and good oral hygiene patients. The specific enzyme activities were measured after 0 (control), 1 and 2 weeks of storage treated with 2.5 mM ethylenediaminetetraacetic acid, 10% (v/v) glycerol or 15% (w/v) polyethylene glycol at three different temperatures, i.e., room temperature, 4 and -20°C. Enzyme activity (unit mL-1) was based on the rate of Nicotinamide Adenine Dinucleotide oxidations standardized at 30°C. The rate of oxidation directly proportional to enzyme’s activity was measured at 340 nm. The specific activity (unit mg-1) was determined through estimated protein content using Bradford analysis. The data were statistically analyzed with paired t-test based on average percentage of enzyme activities from three independent experiments. After two weeks, saliva sample in the presence of polyethylene glycol showed no significant different (p>0.01) at all three temperatures compared to Lactate dehydrogenase basal activity. Lactate dehydrogenase activity of sample in the presence of ethylenediaminetetraacetic acid remained stable (p>0.01) only after a week at room temperature. On the other hand, glycerol managed to stabilize salivary lactate dehydrogenase activity for two weeks at 4 and -20°C. As conclusion, polyethylene glycol showed as the best additive for salivary lactate dehydrogenase storage whereas, ethylenediaminetetraacetic acid suitable only at room temperature for a week. In addition, glycerol was suitable only in cooler conditions.
  M.R. Nur Akmal , Z.A. Intan Zarina , M.A.W. Rohaya , S. Sahidan , Z.A. Zaidah and Z.A. Shahrul Hisham
  Studies have proven that dental pulp contains cells with stem cell activity. Although mice are widely used as a model organism, murine dental pulp stem cells (mDPSCs) are still poorly characterized. In this study, we aim to provide a modified and cost-effective method for isolating mDPSCs and to characterize the isolated cells using molecular markers. Mice incisors’ dental pulps were digested in collagenase 1A and the cell suspensions cultured in α-MEM supplemented with 20% (v/v) fetal bovine serum and 1% (v/v) penicillin-streptomycin. The morphology of the cells was observed and passage 4 cells were subjected to molecular characterization using RT-PCR. We found that the freshly isolated dental pulp cells through enzymatic dissociation method showed a heterogeneous cell morphology which became more homogeneous following cell passage. Semi-quantitative RT-PCR showed analysis that the murine incisors’ DPSCs are Cd105+, Cd13+, Cd29+, Cd146+, Cd166+, Cd90low, Cd73low, Cd105¯ and Cd34¯. Based on these findings, we concluded that dental pulp stem cells from murine incisor could be easily isolated through the method described and the incisor’s dental pulp could be a source of mesenchymal stem cells. Further studies are required to explore the differentiation potential of the isolated mDPSC in vitro.
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