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  1. Reddy LJ, Kumar PS, Pandrangi SL, Chikati R, Srinivasulu C, John A, et al.
    Appl Biochem Biotechnol, 2023 Apr;195(4):2743-2766.
    PMID: 36422804 DOI: 10.1007/s12010-022-04215-w
    The majority of the Earth's ecosystem is frigid and frozen, which permits a vast range of microbial life forms to thrive by triggering physiological responses that allow them to survive in cold and frozen settings. The apparent biotechnology value of these cold-adapted enzymes has been targeted. Enzymes' market size was around USD 6.3 billion in 2017 and will witness growth at around 6.8% CAGR up to 2024 owing to shifting consumer preferences towards packaged and processed foods due to the rising awareness pertaining to food safety and security reported by Global Market Insights (Report ID-GMI 743). Various firms are looking for innovative psychrophilic enzymes in order to construct more effective biochemical pathways with shorter reaction times, use less energy, and are ecologically acceptable. D-Galactosidase catalyzes the hydrolysis of the glycosidic oxygen link between the terminal non-reducing D-galactoside unit and the glycoside molecule. At refrigerated temperature, the stable structure of psychrophile enzymes adjusts for the reduced kinetic energy. It may be beneficial in a wide variety of activities such as pasteurization of food, conversion of biomass, biological role of biomolecules, ambient biosensors, and phytoremediation. Recently, psychrophile enzymes are also used in claning the contact lens. β-D-Galactosidases have been identified and extracted from yeasts, fungi, bacteria, and plants. Conventional (hydrolyzing activity) and nonconventional (non-hydrolytic activity) applications are available for these enzymes due to its transgalactosylation activity which produce high value-added oligosaccharides. This review content will offer new perspectives on cold-active β-galactosidases, their source, structure, stability, and application.
    Matched MeSH terms: beta-Galactosidase/metabolism
  2. Balasubramaniam S, Lee HC, Lazan H, Othman R, Ali ZM
    Phytochemistry, 2005 Jan;66(2):153-63.
    PMID: 15652572
    beta-Galactosidase (EC. 3.2.1.23) from ripe carambola (Averrhoa carambola L. cv. B10) fruit was fractionated through a combination of ion exchange and gel filtration chromatography into four isoforms, viz. beta-galactosidase I, II, III and IV. This beta-galactosidases had apparent native molecular masses of 84, 77, 58 and 130 kDa, respectively. beta-Galactosidase I, the predominant isoform, was purified to electrophoretic homogeneity; analysis of the protein by SDS-PAGE revealed two subunits with molecular masses of 48 and 36 kDa. N-terminal amino acid sequence of the respective polypeptides shared high similarities albeit at different domains, with the deduced amino acid sequence of certain plant beta-galactosidases, thus, explaining the observed low similarity between the two subunits. beta-Galactosidase I was probably a heterodimer that have glycoprotein properties and a pI value of 7.2, with one of the potential glycosylation sites appeared to reside within the 48-kDa-polypeptide. The purified beta-galactosidase I was substantially active in hydrolyzing (1-->4)beta-linked spruce and a mixture of (1-->3)beta- and (1-->6)beta-linked gum arabic galactans. This isoform also had the capability to solubilize and depolymerize structurally intact pectins as well as to modify alkaline-soluble hemicelluloses, reflecting in part changes that occur during ripening.
    Matched MeSH terms: beta-Galactosidase/isolation & purification*; beta-Galactosidase/metabolism*; beta-Galactosidase/chemistry
  3. Rabbani G, Khan MJ, Ahmad A, Maskat MY, Khan RH
    Colloids Surf B Biointerfaces, 2014 Nov 1;123:96-105.
    PMID: 25260221 DOI: 10.1016/j.colsurfb.2014.08.035
    The primary objective of this study is to explore the interaction of β-galactosidase with copper oxide nanoparticles (CuO NPs). Steady-state absorption, fluorescence and circular dichroism (CD) spectroscopic techniques have been employed to unveil the conformational changes of β-galactosidase induced by the binding of CuO NPs. Temperature dependent fluorescence quenching results indicates a static quenching mechanism in the present case. The binding thermodynamic parameters delineate the predominant role of H-bonding and van der Waals forces between β-galactosidase and CuO NPs binding process. The binding was studied by isothermal titration calorimetry (ITC) and the result revealed that the complexation is enthalpy driven, the ΔH°<0, ΔS°<0 indicates the formation of hydrogen bonds between β-galactosidase and CuO NPs occurs. Disruption of the native conformation of the protein upon binding with CuO NPs is reflected through a reduced functionality (in terms of hydrolase activity) of the protein CuO NPs conjugate system in comparison to the native protein and CuO NPs exhibited a competitive mode of inhibition. This also supports the general belief that H-bond formation occurs with NPs is associated with a lesser extent of modification in the native structure. Morphological features and size distributions were investigated using transmission electron microscopy (TEM) and dynamic light scattering (DLS). Additionally the considerable increase in the Rh following the addition of CuO NPs accounts for the unfolding of β-galactosidase. Chemical and thermal unfolding of β-galactosidase, when carried out in the presence of CuO NPs, also indicated a small perturbation in the protein structure. These alterations in functional activity of nanoparticle bound β-galactosidase which will have important consequences should be taken into consideration while using nanoparticles for diagnostic and therapeutic purposes.
    Matched MeSH terms: beta-Galactosidase/chemistry*
  4. Lazan H, Ng SY, Goh LY, Ali ZM
    Plant Physiol Biochem, 2004 Dec;42(11):847-53.
    PMID: 15694277
    The potential significance of the previously reported papaya (Carica papaya L.) beta-galactosidase/galactanase (beta-d-galactoside galactohydrolase; EC 3.2.1.23) isoforms, beta-gal I, II and III, as softening enzymes during ripening was evaluated for hydrolysis of pectins while still structurally attached to unripe fruit cell wall, and hemicelluloses that were already solubilized in 4 M alkali. The enzymes were capable of differentially hydrolyzing the cell wall as evidenced by increased pectin solubility, pectin depolymerization, and degradation of the alkali-soluble hemicelluloses (ASH). This enzyme catalyzed in vitro changes to the cell walls reflecting in part the changes that occur in situ during ripening. beta-Galactosidase II was most effective in hydrolyzing pectin, followed by beta-gal III and I. The reverse appeared to be true with respect to the hemicelluloses. Hemicellulose, which was already released from any architectural constraints, seemed to be hydrolyzed more extensively than the pectins. The ability of the beta-galactanases to markedly hydrolyze pectin and hemicellulose suggests that galactans provide a structural cross-linkage between the cell wall components. Collectively, the results support the case for a functional relevance of the papaya enzymes in softening related changes during ripening.
    Matched MeSH terms: beta-Galactosidase/metabolism*
  5. Boey CC
    J Paediatr Child Health, 2001 Apr;37(2):157-60.
    PMID: 11328471
    OBJECTIVES: To determine the prevalence of lactase deficiency among Malaysian children with recurrent abdominal pain and to describe their clinical characteristics.

    METHODOLOGY: Twenty-four children referred consecutively to the University of Malaya Medical Centre who fulfilled Apley's criteria (at least three episodes of abdominal pain severe enough to affect normal activity over a period longer than 3 months) were tested for lactase deficiency using a pocket breath test analyser (BreatH2 meter; Europa Scientific, Cheshire, England). Lactulose was used to check for hydrogen-producing capacity.

    RESULTS: There were 14 males and 10 females in the study, consisting of five Malays, 14 Chinese and five Indians. Mean age was 9.9 years. Seventeen of the 24 children (70.8%) with recurrent abdominal pain who underwent the breath hydrogen test had a positive result. In those with a negative result, subsequent lactulose administration resulted in a positive rise in breath hydrogen. None of the 24 children developed abdominal pain during the test. All the Indian subjects, 71.4% of the Chinese subjects and 40% of the Malay subjects with recurrent abdominal pain had lactase deficiency. The proportion of boys and girls with lactase deficiency was similar (71.4 vs 70.0%, respectively). There was no significant difference between lactase sufficient and deficient children with recurrent abdominal pain with regard to sex, age, ethnic group and clinical features. Following a lactose-free diet, none of the children in the breath hydrogen positive and negative groups reported any appreciable difference in pain symptoms.

    CONCLUSIONS: The prevalence of lactase deficiency among this group of Malaysian children with recurrent abdominal pain was high, but lactase deficiency did not appear to play an important role in causing the symptoms.

    Matched MeSH terms: beta-Galactosidase/administration & dosage; beta-Galactosidase/deficiency*
  6. Ali ZM, Armugam S, Lazan H
    Phytochemistry, 1995 Mar;38(5):1109-14.
    PMID: 7766393
    The fruit extracts of ripening cv. Harumanis mango contained a number of glycosidases and glycanases. Among the glycosidases, beta-D-galactosidase (EC 3.2.1.23) appeared to be the most significant. The enzyme activity increased in parallel with increase in tissue softness during ripening. Mango beta-galactosidase was fractionated into three isoforms, viz. beta-galactosidase I, II and III by a combination of chromatographic procedures on DEAE-Sepharose CL-6B, CM-Sepharose and Sephacryl S-200 columns. Apparent Km values for the respective beta-galactosidase isoforms for p-nitrophenyl beta-D-galactoside were 3.7, 3.3 and 2.7 mM, and their Vmax values were 209, 1024 and 62 nkat mg-1 protein. Optimum activity occurred at ca pH 3.2 for beta-galactosidase I and II, and pH 3.6 for beta-galactosidase III. Mango beta-galactosidase and its isoforms have galactanase activity, and the activity of the latter in the crude extracts generally increased during ripening. The close correlation between changes in beta-galactosidase activity, tissue softness, and increased pectin solubility and degradation suggests that beta-galactosidase might play an important role in cell wall pectin modification and softening of mango fruit during ripening.
    Matched MeSH terms: beta-Galactosidase/isolation & purification; beta-Galactosidase/physiology*
  7. Makpol S, Yeoh TW, Ruslam FA, Arifin KT, Yusof YA
    PMID: 23948056 DOI: 10.1186/1472-6882-13-210
    Human diploid fibroblasts (HDFs) undergo a limited number of cellular divisions in culture and progressively reach a state of irreversible growth arrest, a process termed cellular ageing. Even though beneficial effects of Piper betle, Chlorella vulgaris and tocotrienol-rich fraction (TRF) have been reported, ongoing studies in relation to ageing is of interest to determine possible protective effects that may reverse the effect of ageing. The aim of this study was to evaluate the effect of P. betle, C. vulgaris and TRF in preventing cellular ageing of HDFs by determining the activity of antioxidant enzymes viz.; catalase, superoxide dismutase (SOD) and glutathione peroxidase.
    Matched MeSH terms: beta-Galactosidase/analysis; beta-Galactosidase/metabolism
  8. Abdul Rahim N, Makpol S, Chua KH, Yusof YA, Top GM, Ngah WZ
    Med J Malaysia, 2008 Jul;63 Suppl A:71-2.
    PMID: 19024989
    Stress-induced premature senescence (SIPS) model is in vitro model of cellular aging. In this study, apoptosis was evaluated in SIPS model and in replicative senescent fibroblasts. We also compared the activity of senescence-associated beta-galactosidase (SA-beta gal) as a biomarker of cellular aging. Our results suggested that SIPS model and senescent fibroblasts might share similar mechanism of aging and apoptosis pathway.
    Matched MeSH terms: beta-Galactosidase/biosynthesis*; beta-Galactosidase/drug effects
  9. Lau KL, Ong EB, Zainudin ZF, Samian MR, Ismail A, Najimudin N
    J Gen Appl Microbiol, 2013;59(3):239-44.
    PMID: 23863294
    Matched MeSH terms: beta-Galactosidase/analysis; beta-Galactosidase/genetics
  10. Younis LT, Abu Hassan MI, Taiyeb Ali TB, Bustami TJ
    Asian J Pharm Sci, 2018 Jul;13(4):317-325.
    PMID: 32104405 DOI: 10.1016/j.ajps.2017.12.003
    This study was designed to investigate the effect of 3D TECA hydrogel on the inflammatory-induced senescence marker, and to assess the influence of the gel on the periodontal ligament fibroblasts (PDLFs) migration in wound healing in vitro. PDLFs were cultured with 20 ng/ml TNF-α to induce inflammation in the presence and absence of 50 µM 3D TECA gel for 14 d. The gel effect on the senescence maker secretory associated-β-galactosidase (SA-β-gal) activity was measured by a histochemical staining. Chromatin condensation and DNA synthesis of the cells were assessed by 4',6-diamidino-2-phenylindole and 5-ethynyl-2'-deoxyuridine fluorescent staining respectively. For evaluating fibroblasts migration, scratch wound healing assay and Pro-Plus Imaging software were used. The activity of senescence marker, SA-β-gal, was positive in the samples with TNF-α-induced inflammation. SA-β-gal percentage is suppressed (>65%, P 
    Matched MeSH terms: beta-Galactosidase
  11. Misson M, Du X, Jin B, Zhang H
    Enzyme Microb Technol, 2016 Mar;84:68-77.
    PMID: 26827776 DOI: 10.1016/j.enzmictec.2015.12.008
    Functional nanomaterials have been pursued to assemble nanobiocatalysts since they can provide unique hierarchical nanostructures and localized nanoenvironments for enhancing enzyme specificity, stability and selectivity. Functionalized dendrimer-like hierarchically porous silica nanoparticles (HPSNs) was fabricated for assembling β-galactosidase nanobiocatalysts for bioconversion of lactose to galacto-oligosaccharides (GOS). The nanocarrier was functionalized with amino (NH2) and carboxyl (COOH) groups to facilitate enzyme binding, benchmarking with non-functionalized HPSNs. Successful conjugation of the functional groups was confirmed by FTIR, TGA and zeta potential analysis. HPSNs-NH2 showed 1.8-fold and 1.1-fold higher β-galactosidase adsorption than HPSNs-COOH and HPSNs carriers, respectively, with the highest enzyme adsorption capacity of 328mg/g nanocarrier at an initial enzyme concentration of 8mg/ml. The HPSNs-NH2 and β-galactosidase assembly (HPSNs-NH2-Gal) demonstrated to maintain the highest activity at all tested enzyme concentrations and exhibited activity up to 10 continuous cycles. Importantly, HPSNs-NH2-Gal was simply recycled through centrifugation, overcoming the challenging problems of separating the nanocarrier from the reaction medium. HPSNs-NH2-Gal had distinguished catalytic reaction profiles by favoring transgalactosylation, enhancing GOS production of up to 122g/l in comparison with 56g/l by free β-galactosidase. Furthermore, it generated up to 46g/l GOS at a lower initial lactose concentration while the free counterpart had negligible GOS production as hydrolysis was overwhelmingly dominant in the reaction system. Our research findings show the amino-functionalized HPSNs can selectively promote the enzyme activity of β-galactosidase for transgalactosylation, which is beneficial for GOS production.
    Matched MeSH terms: beta-Galactosidase/metabolism*
  12. Htwe TT, Karim N, Wong J, Jahanfar S, Mansur MA
    Singapore Med J, 2010 Nov;51(11):856-9.
    PMID: 21140111
    INTRODUCTION: Galectin-3 is a member of the beta-galactoside-binding protein family that plays an important role in cell-to-cell adhesion and in cell-to-matrix interaction. Cellular expression of galectin-3 is correlated with cancer aggressiveness and metastasis.
    METHODS: We examined the differential expression of galectin-3 in a collection of 142 cases of thyroid lesions, including 108 cases of papillary thyroid carcinoma (PTC) and 34 cases of follicular carcinoma (FCA). An immunohistochemical method was applied and semiquantitative scoring was performed on the staining intensity of the positive tissue. Scoring was done on cells at the central portion of the tumour foci and on cells at the periphery that were adjacent to the neighbouring normal thyroid tissue matrix.
    RESULTS: A significantly higher expression (p is 0.001) of galectin-3 was observed in the advancing peripheral thyroid cancer cells compared to the centrally located cells that were not in close contact with the neighbouring stromal tissue in cases with PTC compared to those with FCA.
    CONCLUSION: This finding supported the role of galectin-3 in its cell-to-cell adhesion and cell-to-matrix interaction. Galectin-3 is a potential tumour marker for indicating local and distance metastasis, especially in cases with PTC.
    Matched MeSH terms: beta-Galactosidase/metabolism
  13. Zainuddin A, Chua KH, Abdul Rahim N, Makpol S
    BMC Mol. Biol., 2010;11:59.
    PMID: 20707929 DOI: 10.1186/1471-2199-11-59
    Several genes have been used as housekeeping genes and choosing an appropriate reference gene is important for accurate quantitative RNA expression in real time RT-PCR technique. The expression levels of reference genes should remain constant between the cells of different tissues and under different experimental conditions. The purpose of this study was to determine the effect of different experimental treatments on the expression of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA so that the reliability of GAPDH as reference gene for quantitative real time RT-PCR in human diploid fibroblasts (HDFs) can be validated. HDFs in 4 different treatment groups viz; young (passage 4), senescent (passage 30), H2O2-induced oxidative stress and gamma-tocotrienol (GTT)-treated groups were harvested for total RNA extraction. Total RNA concentration and purity were determined prior to GAPDH mRNA quantification. Standard curve of GAPDH expression in serial diluted total RNA, melting curve analysis and agarose gel electrophoresis were used to determine the reliability of GAPDH as reference gene.
    Matched MeSH terms: beta-Galactosidase/genetics; beta-Galactosidase/metabolism
  14. Goh LH, Mohd Said R, Goh KL
    JGH Open, 2018 Dec;2(6):307-310.
    PMID: 30619942 DOI: 10.1002/jgh3.12089
    Background and Aims: There have been few reports on lactase deficiency (LD) and lactose intolerance (LI) in Malaysia, which has a peculiar mix of three distinct major Asian races-Malay, Chinese, and Indian. The aim of this study was to determine the prevalence of LD and LI in a young multiethnic Malaysian population.

    Methods: Lactase activity was measured with a 13CO2 lactose breath test using an infrared spectrometer. Each subject took 25 g of lactose naturally enriched in 13CO2 together with 250 mL of water after an overnight fast. Breath samples were collected at baseline and at 15-min intervals for 180 min. Subjects were asked to report gastrointestinal (GI) symptoms following ingestion of the lactose test meal.

    Results: Of the 248 subjects tested, 216 (87.1%) were lactase deficient. We found no significant differences in the presentation of LD between gender and races. LD was found in 87.5% of males and 86.8% of females (P = 0.975) and in different races: Chinese (88.5%) versus Malay (83.1%) (P = 0.399), Indian (90.5%) versus Malay (P = 0.295), and Chinese versus Indian (P = 0.902). LI was diagnosed in only 49 (19.8%) subjects; 35 patients had diarrhea, while the remainder had at least two other GI symptoms after the lactose meal.

    Conclusion: The prevalence of LD was high in all three major ethnic groups-Malays, Chinese, and Indians. Ironically, the prevalence of LI was low overall.

    Matched MeSH terms: beta-Galactosidase
  15. Misson M, Dai S, Jin B, Chen BH, Zhang H
    J Biotechnol, 2016 Mar 20;222:56-64.
    PMID: 26876609 DOI: 10.1016/j.jbiotec.2016.02.014
    The nanoenvironment of nanobiocatalysts, such as local hydrophobicity, pH and charge density, plays a significant role in optimizing the enzymatic selectivity and specificity. In this study, Kluyveromyces lactis β-galactosidase (Gal) was assembled onto polystyrene nanofibers (PSNFs) to form PSNF-Gal nanobiocatalysts. We proposed that local hydrophobicity on the nanofiber surface could expel water molecules so that the transgalactosylation would be preferable over hydrolysis during the bioconversion of lactose, thus improve the galacto-oligosaccharides (GOS) yield. PSNFs were fabricated by electro-spinning and the operational parameters were optimized to obtain the nanofibers with uniform size and ordered alignment. The resulting nanofibers were functionalized for enzyme immobilization through a chemical oxidation method. The functionalized PSNF improved the enzyme adsorption capacity up to 3100mg/g nanofiber as well as enhanced the enzyme stability with 80% of its original activity. Importantly, the functionalized PSNF-Gal significantly improved the GOS yield and the production rate was up to 110g/l/h in comparison with 37g/l/h by free β-galactosidase. Our research findings demonstrate that the localized nanoenvironment of the PSNF-Gal nanobiocatalysts favour transgalactosylation over hydrolysis in lactose bioconversion.
    Matched MeSH terms: beta-Galactosidase
  16. Ridzuan N, Al Abbar A, Yip WK, Maqbool M, Ramasamy R
    Stem Cells Int, 2016;2016:8487264.
    PMID: 27579045 DOI: 10.1155/2016/8487264
    The present study is aimed at optimizing the in vitro culture protocol for generation of rat bone marrow- (BM-) derived mesenchymal stem cells (MSCs) and characterizing the culture-mediated cellular senescence. The initial phase of generation and characterization was conducted using the adherent cells from Sprague Dawley (SD) rat's BM via morphological analysis, growth kinetics, colony forming unit capacity, immunophenotyping, and mesodermal lineage differentiation. Mesenchymal stem cells were successfully generated and characterized as delineated by the expressions of CD90.1, CD44H, CD29, and CD71 and lack of CD11b/c and CD45 markers. Upon induction, rBM-MSCs differentiated into osteocytes and adipocytes and expressed osteocytes and adipocytes genes. However, a decline in cell growth was observed at passage 4 onwards and it was further deciphered through apoptosis, cell cycle, and senescence assays. Despite the enhanced cell viability at later passages (P4-5), the expression of senescence marker, β-galactosidase, was significantly increased at passage 5. Furthermore, the cell cycle analysis has confirmed the in vitro culture-mediated cellular senescence where cells were arrested at the G0/G1 phase of cell cycle. Although the currently optimized protocols had successfully yielded rBM-MSCs, the culture-mediated cellular senescence limits the growth of rBM-MSCs and its potential use in rat-based MSC research.
    Matched MeSH terms: beta-Galactosidase
  17. Mohd Sahardi NFN, Jaafar F, Mad Nordin MF, Makpol S
    PMID: 32419792 DOI: 10.1155/2020/1787342
    Background: Ageing resulted in a progressive loss of muscle mass and strength. Increased oxidative stress in ageing affects the capacity of the myoblast to differentiate leading to impairment of muscle regeneration. Zingiber officinale Roscoe (ginger) has potential benefits in reversing muscle ageing due to its antioxidant property. This study aimed to determine the effect of ginger in the prevention of cellular senescence and promotion of muscle regeneration.

    Methods: Myoblast cells were cultured into young and senescent state before treated with different concentrations of ginger standardised extracts containing different concentrations of 6-gingerol and 6-shogaol. Analysis on cellular morphology and myogenic purity was carried out besides determination of SA-β-galactosidase expression and cell cycle profile. Myoblast differentiation was quantitated by determining the fusion index, maturation index, and myotube size.

    Results: Treatment with ginger extracts resulted in improvement of cellular morphology of senescent myoblasts which resembled the morphology of young myoblasts. Our results also showed that ginger treatment caused a significant reduction in SA-β-galactosidase expression on senescent myoblasts indicating prevention of cellular senescence, while cell cycle analysis showed a significant increase in the percentage of cells in the G0/G1 phase and reduction in the S-phase cells. Increased myoblast regenerative capacity was observed as shown by the increased number of nuclei per myotube, fusion index, and maturation index.

    Conclusions: Ginger extracts exerted their potency in promoting muscle regeneration as indicated by prevention of cellular senescence and promotion of myoblast regenerative capacity.

    Matched MeSH terms: beta-Galactosidase
  18. Lim JJ, Ngah WZ, Mouly V, Abdul Karim N
    Oxid Med Cell Longev, 2013;2013:978101.
    PMID: 24349615 DOI: 10.1155/2013/978101
    Skeletal muscle satellite cells are heavily involved in the regeneration of skeletal muscle in response to the aging-related deterioration of the skeletal muscle mass, strength, and regenerative capacity, termed as sarcopenia. This study focused on the effect of tocotrienol rich fraction (TRF) on regenerative capacity of myoblasts in stress-induced premature senescence (SIPS). The myoblasts was grouped as young control, SIPS-induced, TRF control, TRF pretreatment, and TRF posttreatment. Optimum dose of TRF, morphological observation, activity of senescence-associated β-galactosidase (SA-β-galactosidase), and cell proliferation were determined. 50 μg/mL TRF treatment exhibited the highest cell proliferation capacity. SIPS-induced myoblasts exhibit large flattened cells and prominent intermediate filaments (senescent-like morphology). The activity of SA-β-galactosidase was significantly increased, but the proliferation capacity was significantly reduced as compared to young control. The activity of SA-β-galactosidase was significantly reduced and cell proliferation was significantly increased in the posttreatment group whereas there was no significant difference in SA-β-galactosidase activity and proliferation capacity of pretreatment group as compared to SIPS-induced myoblasts. Based on the data, we hypothesized that TRF may reverse the myoblasts aging through replenishing the regenerative capacity of the cells. However, further investigation on the mechanism of TRF in reversing the myoblast aging is needed.
    Matched MeSH terms: beta-Galactosidase/metabolism
  19. Safwani WK, Makpol S, Sathapan S, Chua KH
    Appl Biochem Biotechnol, 2012 Apr;166(8):2101-13.
    PMID: 22391697 DOI: 10.1007/s12010-012-9637-4
    Human adipose-derived stem cells (ASCs) have generated a great deal of excitement in regenerative medicine. However, their safety and efficacy issue remain a major concern especially after long-term in vitro expansion. The aim of this study was to investigate the fundamental changes of ASCs in long-term culture by studying the morphological feature, growth kinetic, surface marker expressions, expression level of the senescence-associated genes, cell cycle distribution and ß-galactosidase activity. Human ASCs were harvested from lipoaspirate obtained from 6 patients. All the parameters mentioned above were measured at P5, P10, P15 and P20. Data were subjected to one-way analysis of variance with a Tukey post hoc test to determine significance difference (P < 0.05). The data showed that growth of ASCs reduced in long-term culture and the ß-galactosidase activity was significantly increased at later passage (P20). The morphology of ASCs in long-term culture showed the manifestation of senescent feature at P15 and P20. Significant alteration in the senescence-associated genes expression levels was observed in MMP1, p21, Rb and Cyclin D1 at P15 and P20. Significant increase in CD45 and HLA DR DQ DP surface marker was observed at P20. While cell cycle analysis showed significant decrease in percentage of ASCs at S and G2/M phase at later passage (P15). Our data showed ASCs cultured beyond P10 favours the senescence pathway and its clinical usage in cell-based therapy may be limited.
    Matched MeSH terms: beta-Galactosidase/metabolism
  20. Makpol S, Durani LW, Chua KH, Mohd Yusof YA, Ngah WZ
    J Biomed Biotechnol, 2011;2011:506171.
    PMID: 21541185 DOI: 10.1155/2011/506171
    This study determined the molecular mechanisms of tocotrienol-rich fraction (TRF) in preventing cellular senescence of human diploid fibroblasts (HDFs). Primary culture of HDFs at various passages were incubated with 0.5 mg/mL TRF for 24 h. Telomere shortening with decreased telomerase activity was observed in senescent HDFs while the levels of damaged DNA and number of cells in G(0)/G(1) phase were increased and S phase cells were decreased. Incubation with TRF reversed the morphology of senescent HDFs to resemble that of young cells with decreased activity of SA-β-gal, damaged DNA, and cells in G(0)/G(1) phase while cells in the S phase were increased. Elongated telomere length and restoration of telomerase activity were observed in TRF-treated senescent HDFs. These findings confirmed the ability of tocotrienol-rich fraction in preventing HDFs cellular ageing by restoring telomere length and telomerase activity, reducing damaged DNA, and reversing cell cycle arrest associated with senescence.
    Matched MeSH terms: beta-Galactosidase/metabolism
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