Displaying publications 61 - 80 of 226 in total

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  1. Isolation, screening, and identification of potential cellulolytic and xylanolytic producers for biodegradation of untreated oil palm trunk and its application in saccharification of lemongrass leaves
    Ang SK, Yahya A, Abd Aziz S, Md Salleh M
    Prep Biochem Biotechnol, 2015;45(3):279-305.
    PMID: 24960316 DOI: 10.1080/10826068.2014.923443
    This study presents the isolation and screening of fungi with excellent ability to degrade untreated oil palm trunk (OPT) in a solid-state fermentation system (SSF). Qualitative assay of cellulases and xylanase indicates notable secretion of both enzymes by 12 fungal strains from a laboratory collection and 5 strains isolated from a contaminated wooden board. High production of these enzymes was subsequently quantified in OPT in SSF. Aspergillus fumigates SK1 isolated from cow dung gives the highest xylanolytic activity (648.448 U g(-1)), generally high cellulolytic activities (CMCase: 48.006, FPase: 6.860, beta-glucosidase: 16.328 U g(-1)) and moderate lignin peroxidase activity (4.820 U/g), and highest xylanolytic activity. The xylanase encoding gene of Aspergillus fumigates SK1 was screened using polymerase chain reaction by a pair of degenerate primers. Through multiple alignment of the SK1 strain's xylanase nucleotide sequences with other published xylanases, it was confirmed that the gene belonged to the xylanase glycoside hydrolase family 11 (GH11) with a protein size of 24.49 kD. Saccharification of lemongrass leaves using crude cellulases and xylanase gives the maximum reducing sugars production of 6.84 g/L with glucose as the major end product and traces of phenylpropanic compounds (vanillic acid, p-coumaric acid, and ferulic acid).
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  2. Statistical optimization of green fluorescent protein production from Escherichia coli BL21(DE3)
    Chew FN, Tan WS, Boo HC, Tey BT
    Prep Biochem Biotechnol, 2012;42(6):535-50.
    PMID: 23030465 DOI: 10.1080/10826068.2012.660903
    An optimized cultivation condition is needed to maximize the functional green fluorescent protein (GFP) production. Six process variables (agitation rate, temperature, initial medium pH, concentration of inducer, time of induction, and inoculum density) were screened using the fractional factorial design. Three variables (agitation rate, temperature, and time of induction) exerted significant effects on functional GFP production in E. coli shake flask cultivation and were optimized subsequently using the Box-Behnken design. An agitation rate of 206 rpm at 31°C and induction of the protein expression when the cell density (OD(600nm)) reaches 1.04 could enhance the yield of functional GFP production from 0.025 g/L to 0.241 g/L, which is about ninefold higher than the unoptimized conditions. Unoptimized cultivation conditions resulted in protein aggregation and hence reduced the quantity of functional GFP. The model and regression equation based on the shake flask cultivation could be applied to a 2-L bioreactor for maximum functional GFP production.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  3. Bioseparation of phycocyanin from Phormidium tergestinum using an aqueous two-phase system
    Tan JS, Abbasiliasi S, Lalung J, Tam YJ, Murugan P, Lee CK
    Prep Biochem Biotechnol, 2021;51(3):260-266.
    PMID: 32876520 DOI: 10.1080/10826068.2020.1808793
    This study aimed at purification of phycocyanin (PC) from Phormidium tergestinum using an aqueous two-phase system (ATPS) comprised of polyethylene glycol (PEG) and salts. The partitioning efficiency of PC in ATPS and the effect of phase composition, pH, crude loading, and neutral salts on purification factor and yield were investigated. Results showed that PC was selectively partitioned toward bottom phase of the system containing potassium phosphate. Under optimum conditions of 20% (w/w) PEG 4000, 10% (w/w) potassium phosphate, 20% (v/v) crude load at pH 7, with addition of 0.5% (w/w) NaCl, PC from P. tergestinum was partially purified up to 5.34-fold with a yield of 87.8%. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the molecular weight of PC was ∼19 kDa. Results from this study demonstrated ATPS could be used as a potential approach for the purification of PC from P. tergestinum.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  4. Evaluation of extraction methods for the identification of proteins from date palm (Phoenix dactylifera L.) seed and flesh
    Lee HX, Ahmad F, Saad B, Ismail MN
    Prep Biochem Biotechnol, 2017 Nov 26;47(10):998-1007.
    PMID: 28857669 DOI: 10.1080/10826068.2017.1365250
    Date fruits are well known to be very nutritious. Nevertheless, the protein contents of the fruit, particularly the seed and flesh, are still understudied, largely due to their difficult physical characteristics. This study was conducted to compare three different protein extraction methods which were the trichloroacetic acid (TCA)-acetone (TCA-A), phenol (Phe), and TCA-acetone-phenol (TCA-A-Phe), and to perform proteomic analysis on date palm seed and flesh. Phe extraction method showed the highest protein yields for both seed (8.26 mg/g) and flesh (1.57 mg/g). Through sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Phe, and TCA-A-Phe extraction methods were shown to be efficient in removing interfering compounds and gave well-resolved bands over a wide range of molecular weights. Following liquid chromatography-tandem mass spectrometry analysis, about 50-64% of extracted proteins were identified with known functions including those involved in glycolysis, Krebs cycle, defense, and storage. Phe protein extraction method was proven to be the optimal method for date flesh and seed.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel/methods
  5. Fulltext Enhanced production, purification, characterization and mechanism of action of salivaricin 9 lantibiotic produced by Streptococcus salivarius NU10
    Barbour A, Philip K, Muniandy S
    PLoS One, 2013;8(10):e77751.
    PMID: 24147072 DOI: 10.1371/journal.pone.0077751
    BACKGROUND: Lantibiotics are small lanthionine-containing bacteriocins produced by lactic acid bacteria. Salivaricin 9 is a newly discovered lantibiotic produced by Streptococcus salivarius. In this study we present the mechanism of action of salivaricin 9 and some of its properties. Also we developed new methods to produce and purify the lantibiotic from strain NU10.

    METHODOLOGY/PRINCIPAL FINDINGS: Salivaricin 9 was found to be auto-regulated when an induction assay was applied and this finding was used to develop a successful salivaricin 9 production system in liquid medium. A combination of XAD-16 and cation exchange chromatography was used to purify the secondary metabolite which was shown to have a molecular weight of approximately 3000 Da by SDS-PAGE. MALDI-TOF MS analysis indicated the presence of salivaricin 9, a 2560 Da lantibiotic. Salivaricin 9 is a bactericidal molecule targeting the cytoplasmic membrane of sensitive cells. The membrane permeabilization assay showed that salivaricin 9 penetrated the cytoplasmic membrane and induced pore formation which resulted in cell death. The morphological changes of test bacterial strains incubated with salivaricin 9 were visualized using Scanning Electron Microscopy which confirmed a pore forming mechanism of inhibition. Salivaricin 9 retained biological stability when exposed to high temperature (90-100°C) and stayed bioactive at pH ranging 2 to 10. When treated with proteinase K or peptidase, salivaricin 9 lost all antimicrobial activity, while it remained active when treated with lyticase, catalase and certain detergents.

    CONCLUSION: The mechanism of antimicrobial action of a newly discovered lantibiotic salivaricin 9 was elucidated in this study. Salivaricin 9 penetrated the cytoplasmic membrane of its targeted cells and induced pore formation. This project has given new insights on lantibiotic peptides produced by S. salivarius isolated from the oral cavities of Malaysian subjects.

    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  6. Fulltext A High-Yield Two-Hour Protocol for Extraction of Human Hair Shaft Proteins
    Wong SY, Lee CC, Ashrafzadeh A, Junit SM, Abrahim N, Hashim OH
    PLoS One, 2016;11(10):e0164993.
    PMID: 27741315 DOI: 10.1371/journal.pone.0164993
    Proteome analysis of the human hair remains challenging due to the poor solubility of hair proteins and the difficulty in their extraction. In the present study, we have developed a rapid extraction protocol for hair shaft protein using alkaline-based buffer. The new protocol accelerated the procedure by reducing the extraction time from at least a day to less than two hours and showed a protein recovery of 47.3 ± 3.72%. Further analyses of the extracted protein sample through sodium dodecyl sulfate polyacrylamide gel electrophoresis and Quadrupole-time-of-flight mass spectrometry analysis unveiled a total of 60 proteins, including 25 that were not previously reported. Identification of these proteins is anticipated to be crucial in helping to understand the molecular basis of hair for potential applications in the future.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  7. Fulltext Antimicrobial activities of Bacillus velezensis strains isolated from stingless bee products against methicillin-resistant Staphylococcus aureus
    Baharudin MMA, Ngalimat MS, Mohd Shariff F, Balia Yusof ZN, Karim M, Baharum SN, et al.
    PLoS One, 2021;16(5):e0251514.
    PMID: 33974665 DOI: 10.1371/journal.pone.0251514
    Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) have reached epidemic proportions globally. Therefore, there is an urgent need for a continuous supply of antibiotics to combat the problem. In this study, bacteria initially identified as species belonging to the Bacillus amyloliquefaciens operational group were re-identified based on the housekeeping gene, gyrB. Cell-free supernatants (CFS) from the strains were used for antimicrobial tests using the agar well diffusion assay against MRSA and various types of pathogenic bacteria. The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and physicochemical characteristics of the CFS were determined. Based on gyrB sequence analysis, five strains (PD9, B7, PU1, BP1 and L9) were identified as Bacillus velezensis. The CFS of all B. velezensis strains showed broad inhibitory activities against Gram-negative and -positive as well as MRSA strains. Strain PD9 against MRSA ATCC 33742 was chosen for further analysis as it showed the biggest zone of inhibition (21.0 ± 0.4 mm). The MIC and MBC values obtained were 125 μl/ml. The crude antimicrobial extract showed bactericidal activity and was stable at various temperatures (40-80°C), pH (4-12), surfactants (Tween 20, Tween 80, SDS and Triton X-100) and metal ions (MgCI2, NaCI2, ZnNO3 and CuSO4) when tested. However, the crude extract was not stable when treated with proteinase K. All these properties resembled the characteristics of peptides. The antimicrobial compound from the selected strain was purified by using solvent extraction method and silica gel column chromatography. The purified compound was subjected to High Performance Liquid Chromatography which resulted in a single peak of the anti-MRSA compound being detected. The molecular weight of the anti-MRSA compound was determined by using SDS-PAGE and zymogram. The size of the purified antimicrobial peptide was approximately ~ 5 kDa. The antimicrobial peptide produced from B. velezensis strain PD9 is a promising alternative to combat the spread of MRSA infections in the future.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  8. Effect of Gibberellin and Heat Shock on the Lipid Composition of Endoplasmic Reticulum in Barley Aleurone Layers
    Grindstaff KK, Fielding LA, Brodl MR
    Plant Physiol, 1996 Feb;110(2):571-581.
    PMID: 12226205
    The heat-shock responses of barley (Hordeum vulgare L. cv Hi- malaya) aleurone layers incubated with or without gibberellic acid (GA3) were compared. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that heat shock blocked the synthesis and secretion of secretory proteins from GA3-treated layers but not untreated layers. This suppression of secretory protein synthesis has been correlated with changes in endoplasmic reticulum (ER) membranes (F.C. Belanger, M. R. Brodl, T.-h.D. Ho [1986] Proc Natl Acad Sci USA 83: 1354-1358; L. Sticher, A.K. Biswas, D.S. Bush, R.L. Jones [1990] Plant Physiol 92: 506-513). Our secretion data suggested that the ER membranes of aleurone layers incubated without GA3 may be more heat shock tolerant. To investigate this, the lipid profiles of membrane extracts in aleurone layers labeled with [14C]glycerol were examined. Heat shock markedly increased [14C]glycerol incorporation into phosphatidylcholine (PC), and gas chromatography revealed an increase in the amount of saturated fatty acids associated with thin layer chromatography-purified PC in GA3-treated layers. In contrast, aleurone layers incubated without GA3 at normal temperature contained PC-associated fatty acids with a greater degree of saturation than GA3-treated layers. Heat shock modestly increased the degree of fatty acid saturation in untreated aleurone layers. This same trend was noted in fatty acids isolated from ER membranes purified by continuous sucrose density centrifugation. We propose that increased fatty acid saturation may help sustain ER membrane function in heat-shocked aleurone layers incubated in the absence of GA3.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  9. First Report of Hibiscus Chlorotic Ringspot Carmovirus in Tuvalu
    Jones P, Devonshire J, Dabek A, Howells C
    Plant Dis, 1998 May;82(5):591.
    PMID: 30857000 DOI: 10.1094/PDIS.1998.82.5.591C
    In September 1997, plants of Hibiscus manihot (locally called nambele) were observed on Vaitupu Island, Tuvalu, exhibiting an angular leaf mosaic and chlorosis that was not always clearly discernible. Electron microscopy of negatively stained sap from affected leaves revealed the presence of numerous isometric virus particles 28 nm in diameter. Poly-acrylamide gel electrophoresis of purified virus gave a single protein band of Mr 38,000 similar to that of the carmoviruses. Immunosorbent electron microscopy tests with antisera kindly provided by N. Spence showed the virus to be hibiscus chlorotic ringspot carmovirus (HCRSV) (1). This virus is also reported from El Salvador, the U.S., Australia, Thailand, Malaysia, Fiji, the Solomon Islands, and Vanuatu. It is not known how the virus reached Tuvalu but we suspect it was via infected cuttings, which were imported for the production of food supplements to combat acute deficiencies of vitamins A and C in the population. The virus is most likely to have been disseminated throughout the islands and atolls of Tuvalu through infected cuttings. Local spread within fields could occur through contaminated hands and cutting implements because of the ease with which the virus is mechanically transmitted. Reference: (1) H. E.Waterworth et al. Phytopathology 66:570, 1976.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  10. A transient assay to evaluate the expression of polyhydroxybutyrate genes regulated by oil palm mesocarp-specific promoter
    Omidvar V, Siti Nor Akmar A, Marziah M, Maheran AA
    Plant Cell Rep, 2008 Sep;27(9):1451-9.
    PMID: 18563415 DOI: 10.1007/s00299-008-0565-2
    The promoter of the oil palm metallothionein-like gene (MT3-A) demonstrated mesocarp-specific activity in functional analysis using transient expression assay of reporter gene in bombarded oil palm tissue slices. In order to investigate the tissue-specific expression of polyhydroxybutyrate (PHB) biosynthetic pathway genes, a multi-gene construct carrying PHB genes fused to the oil palm MT3-A promoter was co-transferred with a construct carrying GFP reporter gene using microprojectile bombardment targeting the mesocarp and leaf tissues of the oil palm. Transcriptional analysis using RT-PCR revealed successful transcription of all the three phbA, phbB, and phbC genes in transiently transformed mesocarp but not in transiently transformed leaf tissues. Furthermore, all the three expected sizes of PHB-encoded protein products were only detected in transiently transformed mesocarp tissues on a silver stained polyacrylamide gel. Western blot analysis using polyclonal antibody specific for phbB product confirmed successful translation of phbB mRNA transcript into protein product. This study provided valuable information, supporting the future engineering of PHB-producing transgenic palms.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  11. Antimicrobial peptides from different plant sources: Isolation, characterisation, and purification
    Tang SS, Prodhan ZH, Biswas SK, Le CF, Sekaran SD
    Phytochemistry, 2018 Oct;154:94-105.
    PMID: 30031244 DOI: 10.1016/j.phytochem.2018.07.002
    Antimicrobial peptides (AMPs), the self-defence products of organisms, are extensively distributed in plants. They can be classified into several groups, including thionins, defensins, snakins, lipid transfer proteins, glycine-rich proteins, cyclotides and hevein-type proteins. AMPs can be extracted and isolated from different plants and plant organs such as stems, roots, seeds, flowers and leaves. They perform various physiological defensive mechanisms to eliminate viruses, bacteria, fungi and parasites, and so could be used as therapeutic and preservative agents. Research on AMPs has sought to obtain more detailed and reliable information regarding the selection of suitable plant sources and the use of appropriate isolation and purification techniques, as well as examining the mode of action of these peptides. Well-established AMP purification techniques currently used include salt precipitation methods, absorption-desorption, a combination of ion-exchange and reversed-phase C18 solid phase extraction, reversed-phase high-performance liquid chromatography (RP-HPLC), and the sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) method. Beyond these traditional methods, this review aims to highlight new and different approaches to the selection, characterisation, isolation, purification, mode of action and bioactivity assessment of a range of AMPs collected from plant sources. The information gathered will be helpful in the search for novel AMPs distributed in the plant kingdom, as well as providing future directions for the further investigation of AMPs for possible use on humans.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  12. Fulltext Effect of glycerol feed in methanol induction phase for Hepatitis B surface antigen expression in Pichia pastoris strain KM71
    Morvarid, A.R., Zeenathul, N.A., Tam, Y.J., Zuridah, H., Mohd-Azmi, M.L., Azizon, B.O.
    Pertanika Journal of Science & Technology, 2012;20(1):31-42.
    MyJurnal
    This study describes expression of HBs Ag in methylotrophic yeast, Pichia Pastoris under alcohol oxidase promoter. A single copy number of HBs Ag gene was transformed into pichia strain of KM 71, a Muts type, by using pA0815 pichia expression vector. The recombinant was cultivated in a shake flask either using methanol or a mixed feed of glycerol -methanol for induction. The HBs Ag gene integrity was justified using direct PCR method. The expressed products in the soluble cell extracts were analyzed by Western blot, SDS page, Bradford assay and ELISA tests. The recombinant HBs Ag was expressed successfully in Pichia pastoris strain KM71 at a high level of HBs Ag protein expression. Thus, an addition of glycerol in the ratio of glycerol per methanol 1/1 (g g-1) consistently produced 2-fold increment in both biomass accumulation and HBs Ag productivity.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  13. Fulltext Physicochemical characterisation and substrate specificity of purified β-1,6-glucanase from Trichoderma Longibrachiatum
    Muskhazli Mustafa, Nor Azwady Abd. Aziz, Anida Kaimi, Nurul Shafiza Noor, Salifah Hasanah Ahmad Bedawi, Nalisha Ithnin
    Pertanika Journal of Science & Technology, 2009;17(1):-.
    MyJurnal
    The β-1,6-glucanases are ubiquitous enzymes which appear to be implicated in the morphogenesis and have the ability to become virulence factor in plant-fungal symbiotic interaction. To our knowledge, no report on ß-1,6-glucanases purification from Trichoderma longibrachiatum has been made, although it has been proven to have a significant effect as a biocontrol agent for several diseases. Therefore, the aim of this study was to purify β-1,6- glucanase from T. longibrachiatum T28, with an assessment on the physicochemical properties and substrate specificity. β-1,3-glucanase enzyme, from the culture filtrate of T. longibrachiatum T28, was successively purified through precipitation with 80% acetone, followed by anionexchange chromatography on Neobar AQ and chromatofocusing on a Mono P HR 5/20 column. (One β-1,6-glucanase) band at 42kDa in size was purified, as shown by the SDS-PAGE. The physicochemical evaluation showed an optimum pH of 5 and optimum temperature of 50°C for enzyme activity with an ability to maintain 100% enzyme stability. Enzyme activity was slightly reduced by 10-20% in the presence of 20 mM of Zn2+, Ca2+, Co2+, Mg2+, Cu2+, Mn2+ and Fe2+. The highest β-1,6-glucanase hydrolysis activity was obtained on pustulan due to the similarity of β-glucosidic bonds followed by laminarin, glucan and cellulose. Therefore, it can be concluded that the characterization of ß-1,6-glucanase secreted by T. longibrachiatum in term of molecular weight, responsed to selected physicochemical factors and the substrate specificity are approximately identical to other Trichoderma sp.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  14. Intraspecific variation of Taenia taeniaeformis as determined by various criteria
    Azuma H, Okamoto M, Oku Y, Kamiya M
    Parasitol Res, 1995;81(2):103-8.
    PMID: 7731915
    The intraspecific variation of four laboratory-reared isolates of Taenia taeniaformis the SRN and KRN isolates from Norway rats, Rattus norvegicus, captured in Japan and Malaysia, respectively; the BMM isolated from a house mouse, Mus musculus, captured in Belgium; and the ACR isolate from a gray red-backed vole, Clethrionomys rufocanus bedfordiae, captured in Japan was examined by various criteria. Eggs of each of the four isolates were orally inoculated into several species of intermediate host. They were most infective to the rodent species from which the original metacestode of each isolate had been isolated in the field, and only the ACR isolate was infective to the gray red-backed vole. Although little difference was found between the SRN, KRN, and BMM isolates by the other criteria, including the morphology of rostellar hooks, the protein composition of the metacestode, and restriction endonuclease analysis of DNA, the ACR isolate was clearly different from the others. It was considered that the ACR isolate was independent as a strain distinct from the other three isolates.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  15. Characterization of IgE-binding proteins in the salivary glands of Simulium nigrogilvum (Diptera: Simuliidae)
    Hempolchom C, Sookrung N, Srisuka W, Reamtong O, Sakolvaree Y, Chaicumpa W, et al.
    Parasitol Res, 2019 Aug;118(8):2353-2359.
    PMID: 31263951 DOI: 10.1007/s00436-019-06383-x
    Simulium dermatitis is an IgE-mediated skin reaction in animals and humans caused by the bites of black flies. Although Simulium nigrogilvum has been incriminated as the main human-biting black fly species in Thailand, information on its salivary allergens is lacking. Salivary gland extract of S. nigrogilvum females was subjected to sodium dodecylsulfate-polyacrylamide gel electrophoresis, and the separated components were applied onto nitrocellulose membranes for immunoblotting, which was performed by probing the protein blots with sera from 17 individuals who were allergic to the bites of S. nigrogilvum. IgE-reactive protein bands were characterized further by liquid chromatography-mass spectrometry (LC-MS/MS) analysis. Nine protein bands (79, 42, 32, 25, 24, 22, 15, 13, and 11 kDa) were recognized in the serum of the subjects. Four of the nine protein bands (32, 24, 15, and 11 kDa) showed IgE reactivity in all (100%) of the tested sera, and they were identified as salivary secreted antigen 5-related protein, salivary serine protease, erythema protein, and hypothetical secreted protein, respectively. Three other proteins, salivary serine protease (25 kDa), salivary D7 secreted protein (22 kDa), and hypothetical protein (13 kDa), reacted with > 50% of the sera. The relevance of the identified protein bands as allergens needs to be confirmed by using pure recombinant proteins, either in the in vivo skin prick test or in vitro detection of the specific IgE in the serum samples of allergic subjects. This will be useful for the rational design of component-resolved diagnosis and allergen immunotherapy for the allergy mediated by the bites of black flies.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  16. Fulltext Comparative profile of circulating antigenic peptides in CSF, serum & urine from patients with neurocysticercosis diagnosed by immunoblotting
    Sahu PS, Parija S, Kumar D, Jayachandran S, Narayan S
    Parasite Immunol., 2014 Oct;36(10):509-21.
    PMID: 24965663 DOI: 10.1111/pim.12124
    Traditionally serum and/or CSF specimens have been used for detection of either specific antibodies or antigens as a supportive diagnosis of NCC. However, in recent days, much interest has been shown employing noninvasive specimens such as urine. In our study, we identified and compared a profile of circulating antigenic peptides of parasite origin in three different body fluids (CSF, serum and urine) obtained from confirmed NCC cases and control subjects. The circulating antigenic peptides were resolved by SDS-PAGE and subjected to immunoblotting. For confirmation of their origin as parasite somatic or excretory secretory (ES) material, immunoreactivity was tested employing affinity purified polyclonal Taenia solium metacestode anti-somatic or ES antibodies, respectively. Only lower molecular weight antigenic peptides were found circulating in urine in contrast to serum and CSF specimens. Few somatic peptides were identified to be 100% specific for NCC (19·5 kDa in all three specimens; 131, 70 kDa in CSF and serum only; 128 kDa in CSF only). Similarly, the specific ES peptides detected were 32 kDa (in all three specimens), 16·5 kDa (in serum and CSF only), and 15 kDa (urine only). A test format detecting either one or more of these specific peptides would enhance the sensitivity in diagnosis of NCC.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  17. Neuroprotective effects of glucomoringin-isothiocyanate against H2O2-Induced cytotoxicity in neuroblastoma (SH-SY5Y) cells
    Jaafaru MS, Nordin N, Rosli R, Shaari K, Bako HY, Saad N, et al.
    Neurotoxicology, 2019 12;75:89-104.
    PMID: 31521693 DOI: 10.1016/j.neuro.2019.09.008
    Neurodegenerative diseases (NDDs) are pathological conditions characterised by progressive damage of neuronal cells leading to eventual loss of structure and function of the cells. Due to implication of multi-systemic complexities of signalling pathways in NDDs, the causes and preventive mechanisms are not clearly delineated. The study was designed to investigate the potential signalling pathways involved in neuroprotective activities of purely isolated glucomoringin isothiocyanate (GMG-ITC) against H2O2-induced cytotoxicity in neuroblastoma (SH-SY5Y) cells. GMG-ITC was isolated from Moringa oleifera seeds, and confirmed with NMR and LC-MS based methods. Gene expression analysis of phase II detoxifying markers revealed significant increase in the expression of all the genes involved, due to GMG-ITC pre-treatment. GMG-ITC also caused significant decreased in the expression of NF-kB, BACE1, APP and increased the expressions of IkB and MAPT tau genes in the differentiated cells as confirmed by multiplex genetic system analysis. The effect was reflected on the expressed proteins in the differentiated cells, where GMG-ITC caused increased in expression level of Nrf2, SOD-1, NQO1, p52 and c-Rel of nuclear factor erythroid factor 2 (Nrf2) and nuclear factor kappa-B (NF-kB) pathways respectively. The findings revealed the potential of GMG-ITC to abrogate oxidative stress-induced neurodegeneration through Nrf2 and NF-kB signalling pathways.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  18. Biochemical and radical-scavenging properties of sea cucumber (Stichopus vastus) collagen hydrolysates
    Abedin MZ, Karim AA, Latiff AA, Gan CY, Ghazali FC, Barzideh Z, et al.
    Nat Prod Res, 2014;28(16):1302-5.
    PMID: 24670209 DOI: 10.1080/14786419.2014.900617
    The molecular mass distribution, amino acid composition and radical-scavenging activity of collagen hydrolysates prepared from collagen isolated from the sea cucumber Stichopus vastus were investigated. β and α1 chains of the collagen were successfully hydrolysed by trypsin. The molecular mass distribution of the hydrolysates ranged from 5 to 25 kDa, and they were rich in glycine, alanine, glutamate, proline and hydroxyproline residues. The hydrolysates exhibited excellent radical-scavenging activity. These results indicate that collagen hydrolysates from S. vastus can be used as a functional ingredient in food and nutraceutical products.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  19. Plant proteins, minerals and trace elements of Eurycoma longifolia (Tongkat Ali)
    Chua LS, Abdul-Rahman N, Rosidi B, Lee CT
    Nat Prod Res, 2013 Mar;27(4-5):314-8.
    PMID: 22468741 DOI: 10.1080/14786419.2012.676552
    A water extraction method has been used to extract plant proteins from the roots of Eurycoma longifolia harvested from Perak and Pahang, Malaysia. On the basis of the spectroscopic Bradford assay, Tongkat Ali Perak and Pahang contained 0.3868 and 0.9573 mg mL(-1) of crude protein, respectively. The crude proteins were separated by one dimensional 15% sodium dodecyl sulphate polyacrylamide gel electrophoresis into two (49.8 and 5.5 kD) and four (49.8, 24.7, 21.1 and 5.5 kD) protein spots for Tongkat Ali Perak and Pahang, respectively. Isoleucine was present in the highest concentration significantly. Both plant samples showed differences in the mineral and trace element profiles, but the minerals calcium, magnesium and potassium were present in the highest concentration. The highly concerned toxic metals such as arsenic and lead were not detected.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  20. Detection of IgM and IgG antibodies to Cryptococcus neoformans proteins in blood donors and HIV patients with active cryptococcosis
    Chai HC, Tay ST
    Mycoses, 2009 Mar;52(2):166-70.
    PMID: 18643920 DOI: 10.1111/j.1439-0507.2008.01549.x
    The serological responses to Cryptococcus neoformans proteins of blood donors and HIV patients with active cryptococcosis from a tropical region were investigated in this study. Exposure to C. neoformans, an organism ubiquitous in the environment, contributes to the antibody responses observed in the blood donors. IgG responses to cryptococcal proteins were stronger than IgM responses in most sera tested in this study. A 53-kDa cryptococcal protein fragment was identified as the most immunoreactive protein on the IgM immunoblots of both blood donors and patients. Overall, there was no obvious difference in IgG responses of patients when compared with blood donors. Some immunogenic protein fragments (27.5, 76, 78 and 91.5 kDa) were detected at least two times more frequently on IgM immunoblots of patients compared with those of blood donors. It is yet to be investigated whether the proteins identified in this study may have any potential to be used as biomarker for cryptococcosis.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
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