Fungi is known to produce a wide range of biologically active metabolites and enzymes. Enzymes produced by fungi are utilized in food and pharmaceutical industries because of their rich enzymatic profile. Filamentous fungi are particularly interesting due to their high production of extracellular enzymes which has a large industrial potential. The aim of this study is to isolate potential soil fungi species that are able to produce functional enzymes for industries. Five Aspergillus species were successfully isolated from antibiotic overexposed soil (GPS coordinate of N3.093219 E101.40269) by standard microbiological method. The isolated fungi were identified via morphological observations and molecular tools; polymerase chain reactions, ITS 1 (5’- TCC GTA GGT GAA CCT GCG G3’) forward primer and ITS 4 (5’-TCC TCC GCT TAT TGA TAT GC-3’) reverse primer. The isolated fungi were identified as Aspergillus sydowii strain SCAU066, Aspergillus tamarii isolate TN-7, Aspergillus candidus strain KUFA 0062, Aspergillus versicolor isolate BAB-6580, and Aspergillus protuberus strain KAS 6024. Supernatant obtained via submerged fermentation of the isolated fungi in potato dextrose broth (PDB) and extracted via centrifugation was loaded onto specific media to screen for the production of xylanolytic, cellulolytic and amylolytic enzymes. The present findings indicate that Aspergillus sydowii strain SCAU066 and Aspergillus versicolor isolate BAB-6580 have great potential as an alternative source of xylanolytic, cellulolytic and amylolytic enzymes.
Kajian ini dilakukan bagi melihat keupayaan sel mononukleus sistem darah pusat membeza kepada sel osteoblas dan osteoklas secara in vitro bagi tiga tempoh proliferasi yang berbeza. Sel mononukleus sistem darah pusat dikulturkan di dalam medium pemilihan proliferasi bagi tiga tempoh proliferasi yang berbeza iaitu jangkamasa pendek (5 hari), sederhana (15 hari) dan panjang (30 hari). Keupayaan sel mononukleus untuk membeza kepada sel osteoblas dan osteoklas seterusnya diperhatikan pada setiap jenis sel ini. Medium proliferasi ditambah dengan faktor pembezaan asid askorbik dan β-gliserofosfat bagi membezakan sel mononukleus kepada sel osteoblas. Bagi pembezaan sel osteoklas pula, RANKL dan M-CSF ditambah ke dalam medium proliferasi. Bagi kawalan, sel yang sama digunakan tanpa penambahan faktor pembezaan. Viabiliti sel yang membeza daripada sel jangkamasa pendek, sederhana dan panjang menunjukkan sel-sel tersebut berupaya untuk bermandiri tanpa sebarang peningkatan yang signifikan sehingga 10 dan 14 hari dengan kehadiran faktor-faktor pembezaan tertentu di dalam medium pembezaan masing-masing. Analisis biokimia ke atas aktiviti alkali fosfatase (ALP) dan asid fosfatase rintang tartarat (TRAP) menunjukkan peningkatan yang signifikan (p<0.05) apabila dikulturkan di dalam medium pembezaan masing-masing. Kesimpulannya, keupayaan sel primitif untuk membeza kepada sel osteoblas dan osteoklas matang adalah hampir sama bagi ketiga-tiga jenis jangkamasa proliferasi tetapi mempunyai kadar proliferasi yang berlainan iaitu 0.37, 0.55 dan 0.72 pembahagian/hari masing-masing bagi sel jangkamasa pendek, sederhana dan panjang. Sel mononukleus yang diasingkan daripada darah periferi ini sangat primitif kerana berpotensi untuk membeza kepada dua jenis sel matang yang berasal daripada sel stem yang berbeza, justeru boleh dikategorikan sebagai sel stem multipoten.
Aspergillus sp. is an extremely resilient species that can be found everywhere in the environment and
is present abundantly in water and soil. The defining characteristic of Aspergillus sp. is their extensive
hyphal network which enable them to survive anywhere, even in very harsh conditions. This study was
carried out to isolate the filamentous fungi from peat soil of animal agricultural farm and characterise
them based on their morphological and molecular characteristics. Growth rate of each isolated fungi was
also evaluated in order to determine the period of maturity for each fungi. Soil samples were collected,
weighed and then dissolved in sterile distilled water. The samples were serially diluted and spread
onto potato dextrose agar (PDA) for isolation. Different isolated colonies that were morphologically
different from each plate was purified and sub-cultured onto new media for macroscopic and microscopic
identifications. For molecular identification, a conventional technique was used in genomic DNA
extraction of filamentous fungi due to their thick cell wall and presence of surface proteins protecting the
fungus. These characteristics make it difficult to harvest the genomic DNA. Polymerase chain reaction
(PCR) was carried out using internal transcribed spacer primers; ITS1 (forward) and ITS4 (reverse).
The morphological identification and molecular
technique showed that majority of these isolated
fungi are Aspergillus sp.
The evolution of cosmetic products results in the growing demands for cosmetics that are preservatives free. Plant essential oils were found to be a promising antimicrobial and also antioxidant agent. In this study, Cymbopogon citratus (lemongrass), Laurus nobilis (bay leaf) and Backhousia citriodora (lemon myrtle) essential oils were selected and evaluated for their antimicrobial properties. It was found that Laurus nobilis exhibited strong antimicrobial activity against the selected bacteria Streptococcus saprophyticus (ATCC 49619), Streptococcus aureus (ATCC 22923), Streptococcus pyogenes (ATCC 29436), Pseudomonas aeruginosa (ATCC 13048), Klebsiella pneumoniae (ATCC 700603), Escherichia coli (ATCC 22922) with MIC ranging between 7.8 ug/mL to 250 μg/mL. The antioxidant activity of selected essential oils was determined by antioxidant assays which were 1,1-Diphenyl-2-picrylhydrazyl assay (DPPH), determination of ferric reducing antioxidant power assay (FRAP) and β-Carotene/linoleic acid bleaching assay. Backhousia citriodora and Laurus nobilis showed the highest antioxidant activity.
n-Octanal and β-Selinene were identified to be the major components with peak area of 26.37 % and 13.92 % respectively in secondary metabolites analysis by Gas Chromatography-Mass Spectrometry (GCMS).
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.
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].
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.
Air liur berpotensi menjadi punca DNA yang mudah diambil bagi kajian klinikal kerana tidak invasif berbanding sampel darah. Kajian ini dijalankan untuk memencilkan dan menulenkan DNA genom daripada sampel air liur manusia serta mengkaji kesan penyimpanan terhadap kualiti DNA genom. Sampel air liur (n=5) disimpan dalam penimbal Tris-NaCl EDTA (TNE) pada suhu bilik (25°C) mengikut tempoh masa yang ditetapkan iaitu, segar (tanpa penyimpanan), 1,2,3 dan 4 bulan. Pemencilan dan penulenan DNA dilakukan menggunakan kaedah fenol-kloroform. Seterusnya, PCR telah dijalankan untuk mengetahui ketulenan DNA yang diekstrak menggunakan amplifikasi pada kawasan jujukan beta-globin dan mengenal pasti kehadiran bakteria melalui jujukan yang mengekod 16S rDNA. Keputusan menunjukkan fragmen DNA gen beta-globin manusia hanya berjaya diamplifikasi daripada sampel segar. Sampel air liur yang disimpan dalam penimbal TNE pada suhu bilik tidak mampu menstabilkan DNA genom manusia untuk jangka masa lama dan hanya berkesan untuk tempoh yang singkat iaitu, kurang daripada 1 bulan. Kesimpulannya, hanya sampel air liur segar sahaja yang berupaya memencil DNA genom.