Sel osteoblas merupakan sel mononukleus yang bertanggungjawab untuk pembentukan tulang. Sel mononukleus telah terbukti mampu membeza kepada sel osteoblas setelah diaruh oleh kombinasi asid askorbik dan ß-gliserofosfat sebagai faktor pembezaan. Tujuan kajian ini adalah bagi melihat kesan aruhan ß-gliserofosfat terhadap ampaian sel mononukleus daripada darah periferi manusia secara in vitro. Sel mononukleus dipencilkan daripada darah periferi manusia dengan menggunakan larutan Ficoll-Paque™ Plus melalui kaedah pengemparan kecerunan ketumpatan. Sel mononukleus kemudian dikultur selama 7 hari di dalam medium proliferasi sebelum diaruh dengan ß-gliserofosfat pada kepekatan 1 mM, 5 mM, 10 mM, 20 mM, dan 100 mM. Pada hari 0, 1, 3, 7, dan 14, penentuan profil aktiviti enzim alkalin fosfatase (ALP) dan analisis morfologi bagi sel osteoblas dilakukan dalam medium masing-masing. Aktiviti enzim ALP dan analisis morfologi menunjukkan peningkatan yang signifikan (p<0.05) antara sel yang diaruh dengan sel kawalan negatif melalui statistik ujian-t berpasangan. Kesimpulannya, kehadiran ß-gliserofosfat sahaja mampu untuk mengaruh pembezaan sel mononukleus kepada sel osteoblas. Kesan ß-gliserofosfat terhadap pembezaan sel mononukleus kepada sel osteoblas menunjukkan profil enzimologi (aktiviti enzim ALP) dan morfologi yang hampir sama dengan kawalan positif (asid askorbik dan ß-gliserofosfat). Berdasarkan kepada analisis enzimologi dan morfologi 1 mM ß-gliserofosfat adalah kepekatan yang paling sesuai untuk pembezaan sel osteoblas secara in vitro.
This study was carried out to determine the optimal parameters for the production of biomass of Trichoderma virens UKMP-1M, a fungus isolated from oil-polluted wastewater. The isolate showed maximum growth at day six after incubation in Mineral Salt Medium (MSM) in the presence of 3% (v/v) heavy Khefji Sour crude oil. Although it grew at pH between 5.0 and 7.0, it grew best at pH 5.5. T. virens UKMP-1M grew at temperatures between 25°C and 35°C, with its highest growth at 30°C. Aeration by agitation at 200 rpm was shown to yield the greatest biomass. Peptone at concentration of 1.5% (w/v) was determined to be a better nitrogen source than urea, potassium nitrate (KNO3), yeast extract, ammonium sulphate ((NH4)2SO4) and ammonium chloride (NH4Cl). Addition of 1% (v/v) crude oil to the MSM medium led to higher biomass production than the addition of 3%, 5%, 7% and 10% (v/v) crude oil. The result also revealed that 40% of total petroleum hydrocarbon (TPH), 100% of pristane and 74% of phytane compounds were degraded after 9 days of incubation at optimal physical and nutrient parameters.
Proses pergerakan gigi semasa rawatan ortodontik boleh dikelaskan kepada empat fasa iaitu pengaktifan (berkait inflamasi terhadap tisu serta kematian sel), penyerapan, proses berbalik dan pembentukan tulang. Pergerakan gigi ini berkait rapat dengan perubahan metabolik di sekitar mulut. Objektif kajian ini adalah untuk menentukan profil penanda biologi enzim di dalam air liur individu yang menerima rawatan ortodontik iaitu laktat dehidrogenas (LDH) bagi proses inflamasi, asid fosfatase rintang tartarat (TRAP) bagi proses penyerapan tulang dan alkali fosfatase (ALP) bagi proses pembentukan tulang. Sampel air liur diambil daripada 6 individu yang menerima rawatan ortodontik. Aktiviti kesemua enzim diambil sebelum pendakap dipasang (aktiviti normal) diikuti dengan hari ke-3, 7, 10, 14, 17, 21, 24, 28 dan 31 selepas pengaktifan. Hasil kajian mendapati kesemua enzim (LDH, TRAP and ALP) menunjukkan peningkatan yang signifikan (p≤0.05) selepas rawatan diberikan berbanding aktiviti normal. LDH didapati meningkat pada peringkat awal rawatan (hari ke-3,7 dan 10), TRAP pada hari ke 14 dan 17 diikuti dengan ALP pada hari ke-17, 21 dan 24. Sebagai kesimpulan, profil enzim sepanjang rawatan ortodontik menunjukkan proses inflamasi berlaku di peringkat awal rawatan diikuti proses penyerapan dan pembentukan tulang. Selain itu, keseluruhan fasa inflamasi, penyerapan dan pembentukan tulang ortodontik didapati mengambil masa 24 hari.
We report here the isolation and sequence analysis of a cDNA fragment for an endoglucanase gene from Aspergillus terreus SUK-I. A pair of primers was designed based on conserved regions of endoglucanase gene. Amplification of A. terreus SUK-I cDNA using the primers produced a DNA band of 642 bp. The cDNA fragment was purified and cloned into pCR®ll-TOPO plasmid vector. The cDNA insert was designated as SA2 and sequenced. Analysis of the SA2 sequence showed a high degree of identity towards endoglucanase gene sequences from A. aculeatus, A kawachii and Talaromyces emersonii. While, the putative amino acid sequence of SA2 showed 74 % identity towards endoglucanase protein from Thermoascus aurantiacus and 69 % identity towards endoglucanase protein from A. kawachii, A. niger, A. aculeatus and A. nidulans. A 66 % identity between SA2 putative amino acid sequence and A. oryzae endoglucanase protein sequence was also observed. Therefore, we concluded that SA2 codes for a putative endoglucanase gene of A. terreus SUK-I.
[Kami melaporkan di sini pemencilan dan analisis satu fragmen cDNA untuk gen endoglukanase daripada Aspergillus terreus SUK-I. Sepasang pencetus telah direkabentuk berasaskan kepada kawasan terpelihara gen endoglukanase. Amplifikasi cDNA A. terreus SUK-I menggunakan pencetus berkenaan menghasilkan satu jalur DNA bersaiz 642 pb. Fragmen cDNA berkenaan telah ditulenkan dan diklonkan ke dalam vektor plasmid pCR®II-TOPO. cDNA selitan telah dilabelkan sebagai SA2 dan dijujukkan. Analisis terhadap jujukan SA2 menunjukkan sebahagian daripada jujukan SA2 mempunyai darjah identiti yang tinggi terhadap jujukan gen endoglukanase daripada A. aculeatus, A. kawachii dan Talaromyces emersonii. Sementara, jujukan asid amino putatif SA2 menunjukkan identiti sebanyak 74% terhadap protein endoglukanase daripada Thermoascus aurantiacus dan identiti sebanyak 69% terhadap protein endoglukanase daripada A. kawachii, A. niger, A. aculeatus dan A. nidulans. ldentiti sebanyak 66% juga diperhatikan di antara jujukan asid amino putatif SA2 dan jujukan protein endoglukanase A. oryzae. Oleh itu, kami membuat kesimpulan bahawa SA2 mengkodkan gen putative endoglukanase A. terreus SUK-I].
The isolation method for dental pulp stem cells (DPSCs) is still unclear to obtain a conducive environment for DPSCs to
proliferate. Enzymatic digestion and outgrowth method are two commonly used methods for DPSCs isolation but are not
well characterized in mice DPSCs. This study aimed to compare these isolation methods and differentiation potential
of mice DPSCs into bone cells. Dental pulp was extracted from mice’s incisors and subjected to isolation either by
collagenase 1A or culture of pulp tissue in complete alpha-Modified Eagle Medium (αMEM). Both cells isolated were
cultured until passage 4 and subjected to in vitro proliferation and differentiation analysis. Both cells exhibited fibroblastliked
morphology, but cells isolated by enzyme digestion proliferate faster compare to outgrowth method. After 21 days
of osteoblast differentiation, DPSCs isolated from enzyme digestion method showed alkaline phosphatase (ALP) activity
slightly different as compared to outgrowth method. In conclusion, there is a significant difference between the cells
isolated from enzyme digestion compare to outgrowth method with regard to proliferation and osteoblast differentiation.
Thus, it is preferable to isolate by enzyme digestion as it is faster and consistent compared to outgrowth method.
Stem cell is defined as the ability of the cell to proliferate themselves and differentiate into more than one type of cells. Human mononucleated cell (MN cell) is a suspension cell that was isolated from peripheral blood that was originated from monocyte-machrophage lineage or hematopoietic stem cells. The cells were cultured for 30 days in complete media (CM) which consist of Alpha Minimal Essential Medium (αMEM) with 2% (v/v) Penicillin-Streptomycin and 10% (v/v) Newborn Calf Serum (NBCS). The respective cells were differentiated at day 7 after in vitro proliferation in CM into osteoblastic cells by adding ascorbic acid and β-glycerophosphate. In addition, human recombinant Receptor Activator of Nuclear Factor-β Ligand (hrRANKL) and human recombinant Macrophage-Colony Stimulating Factor (hrM-CSF) were added to induce osteoclastic differentiation of MN cells. Cells that were cultured in CM served as a control and were subjected to the same approach as differentiated cells. The 30 days cultured cells in CM showed a significant increment (p < 0.05) of viable cells compared to day 0 (n=3). The specific activity of Alkaline Phosphatase (ALP) for osteoblast differentiation and Tartrate Resistant Acid Phosphatase (TRAP) for osteoclast differentiation were evaluated via biochemical assay until day 14 and day 10 for osteoblast and osteoclast sample, respectively. ALP and TRAP enzyme showed a significant increment (p < 0.05) after 14 and 10 days of differentiation compared to control cells. As a conclusion, human mononucleated cells are believed to have the potential to be defined as a multipotent stem cell based on their fulfillment of stem cell characteristics.
Kolagen jenis 1 α- 2 adalah salah satu daripada kolagen yang dapat ditemui dalam tisu pulpa gigi. Kolagen jenis 1 membentuk hampir 56% daripada kawasan intraselular di dalam pulpa. Objektif kajian ini adalah untuk memencilkan gen Col1A2 daripada tisu pulpa Orycytolagus cuniculus. Gen penyelenggara, iaitu gen gliseraldehid 3-fosfat dehidrogenase (GAPDH) telah dipilih sebagai gen kawalan positif. Pencetus spesifik bagi kedua-dua gen Col1A2 dan GAPDH telah direkabentuk berdasarkan perisian Primer Premier Versi 5 berdasarkan jujukan yang diperolehi daripada NCBI (National Center for Biotechnology Information) RNA dipencilkan daripada tisu pulpa Oryctolagus cuniculus. Kedua-dua gen telah diamplifikasi dan hasil produk PCR, bersaiz 893 pb (Col1A2) dan 229 pb (GAPDH) telah didapati. Hasil analisis jujukan menunjukkan bahawa klon, Col1A2 mempunyai hampir 99% persamaan manakala GAPDH mempunyai 100% persamaan dengan jujukan yang terdapat dalam pengkalan data NCBI. Ini menunjukkan transkrip Col1A2 berjaya dikesan daripada tisu pulpa O. cuniculus dewasa.
Ancient remains are considered very valuable artefacts, as they allow for the study of ancient cultures, phylogeny, evolution and the reconstruction of demographic history. To obtain all the information contained within remains, the investigation of such samples requires the expertise and various techniques from multiple fields of study. The present review focuses on the molecular biology and radiographic approaches used to identify ancient samples. Studies of ancient remains face various limitations; for example, the quality and quantity of the ancient samples can affect the difficulty of the investigations. Due to these limitations, new sophisticated techniques are being introduced to replace the earlier conventional techniques. A search was conducted using PubMed, Scopus, Science Direct and Science Finder to provide a new and timely review on the molecular mitochondrial DNA and radiographic analysis for human archaeology identification. The present review has determined that molecular biological approaches are very accurate and useful for the use in the ancestral determination of incomplete specimens, whereas observations of the dental pulp chamber are suitable for age at death estimations in both adults and children. However, these techniques are expensive and require expert personnel. Therefore, conventional approaches remain the favourite methods of most institutions, especially in Asia.
Stem cells, also known as mother cells are capable of undergoing both cell division and differentiation. The most primitive stem cells are totipotent cells which are capable of producing a complete organism from one cell. There are two types of haemopoietic stem cells depending on their developmental stages known as embryo and adult haemopoietic stem cells. Studies showed that only 0.01-0.05% of total bone marrow cell population consists of haemopoietic stem cells. This small population of stem cells exists in three different sizes with different characteristics. In addition, the microenvironment which contains various regulatory molecules plays an important role in the differentiation of stem cells into specific adult cells.
[Sel stem juga dikenali sebagai sel induk berupaya untuk menjalani kedua-dua proses pembahagian dan pembezaan sel. Sel stem yang paling primitif iaitu sel totipoten berupaya untuk membentuk satu organisma lengkap daripada satu sel. Sel stem hemopoietik terdiri daripada dua jenis bergantung kepada peringkat perkembangan individu iaitu sel stem hemopoietik embrio dan dewasa. Kajian mendapati hanya 0.01-0.05% daripada keseluruhan populasi sel sumsum tulang berupaya bertindak sebagai sel stem hemopoietik. Daripada julat yang kecil ini sel stem hemopoietik wujud dalam tiga saiz yang mempunyai ciri yang berbeza. Selain daripada itu mikrosekitaran yang mempunyai molekul-molekul regulatori yang berbeza-beza juga memainkan peranan yang penting dalam pembezaan sel stem kepada sel-sel matang yang spesifik].
Penyerapan akar gigi apeks luaran (PAAL) adalah salah satu kesan negatif semasa rawatan ortodontik selain daripada gigi
yang mengalami trauma. Objektif kajian ini adalah untuk melihat hubungan antara sejarah trauma dan kejadian PAAL serta
membandingkan tahap keterukan PAAL antara gigi trauma dan tanpa trauma selepas enam dan 12 bulan rawatan ortodontik.
Sampel kajian merupakan gigi insisor tengah maksila daripada 23 subjek (8 lelaki dan 15 wanita berumur 12 hingga 26
tahun) dengan 19 mempunyai trauma (tanpa penyerapan akar gigi) dan 27 tanpa trauma. Rawatan ortodontik dilakukan
dengan menggunakan dawai arkus NiTi 0.014”, 0.018” dan 0.018” × 0.025” pada enam bulan pertama. Selepas enam
bulan, rawatan ortodontik diteruskan dengan menggunakan dawai arkus keluli tahan karat saiz 0.019” × 0.025” sehingga
rawatan ortodontik mencapai satu tahun. PAAL gigi diukur melalui imej tomografi berkomputer pancaran-kon (CBCT) yang
diambil sebelum (X0
), selepas enam bulan (X6
) dan selepas 12 bulan (X12) rawatan ortodontik. Penyerapan akar dikira
dengan menolak panjang gigi pada X6
dan X12 dengan panjang gigi pada X0
. Kejadian PAAL dalam kumpulan trauma dan
tanpa trauma masing-masing adalah 89.5% dan 77.8% (penyerapan akar rendah dari 2 mm) selepas enam bulan rawatan
ortodontik. Selepas 12 bulan rawatan ortodontik, semua gigi menunjukkan PAAL. Kejadian PAAL antara gigi trauma dan
tanpa trauma tidak menunjukkan perbezaan yang signifikan (p>0.05). Dalam kajian ini, gigi yang mengalami trauma serta
tanpa trauma membentuk PAAL pada kadar yang sama selepas enam dan 12 bulan rawatan orthodontik. Oleh itu, adalah
selamat untuk melakukan rawatan ortodontik kepada pesakit yang mempunyai sejarah trauma pada gigi.
Alkaline phosphatase (ALP), tartrate-resistant acid phosphatase (TRAP) and aspartate aminotransferase (AST) activities were studied as biomarkers of canine movement. Root resorption was also evaluated in canines subjected to the orthodontic forces. Nineteen subjects randomly received 100 and 150 g force using self-ligating brackets (SLB) either on the right or left site of maxillary arch. Gingival crevicular fluid samples were collected at distal sites of canines for five consecutive weeks. The activities of ALP, TRAP and AST were assayed and measured spectrophotometrically. Canine movement was measured for five consecutive weeks while root resorption was monitored at baseline, week 0 and week 5 using periapical radiographs. In 100 g group, TRAP activity significantly increased in week 3-5 when compared to TRAP baseline activity. However, ALP and AST activities slightly increased. In 150 g group, ALP and TRAP activities slightly increased when compared with their baseline activities. However, AST significantly increased in week 5. Canine movement and root resorption were not significantly different (p<0.05) in both groups. A force of 100 and 150 g slightly increased the bone modeling process and resulted in similar canine movement and root resorption. Therefore, 100 g force could be an optimum force for canine retraction and is preferable (compared with 150 g force) in canine retraction using SLB.
Famili Piperaceae pada keseluruhannya terdiri daripada 1,000 hingga 2,000 spesies yang boleh dijumpai di kawasan tropika dan subtropika. Dalam kajian ini, ekstrak etanol digunakan untuk melihat aktiviti sitotoksik ke atas sel kanser hati manusia (HepG2) dan sel bukan kanser hati Chang melalui kaedah pengasaian MTT (3,4 [dimetiltiazol-2-yl]-2, 5-difeniltetrazolium bromida). Sebanyak lapan spesies daripada famili Piperaceae telah terpilih untuk analisis aktiviti antitumor. Hasil kajian mendapati kesemua spesies Piperaceae (P. sarmentosum, P. ramifilum, P. paucistigmum, P. betle, P. macronatum, P. ridleyi, P. magnibaccum dan P. miniatum) menunjukkan aktiviti sitotoksik dengan ekstrak etanol Piper sarmentosum mempunyai nilai bacaan IC50 yang paling rendah ke atas sel HepG2 iaitu 12.5 μg/mL. Tiada aktiviti sitotoksik telah ditunjukkan oleh kesemua ekstrak etanol tumbuhan yang diuji aktiviti sitotoksik ke atas sel Chang kerana nilai bacaan IC50 kesemua ekstrak etanol yang diperlakukan ke atas sel Chang melebihi nilai piawai iaitu 30 μg/mL. Kaedah analisis viabiliti sel menggunakan tripan biru pula mendapati ekstrak etanol P. sarmentosum menurun secara signifikan (p<0.05) terhadap sel HepG2 berbanding kawalan. Kesimpulannya, kaedah MTT menunjukkan kesemua ekstrak etanol famili Piperaceae memberikan aktiviti sitotoksik dan kaedah tripan biru merupakan kaedah alternatif bagi penentuan kesitotoksikan sesuatu ekstrak.