Displaying publications 41 - 60 of 314 in total

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  1. Cheah YT, Lakbir Singh HKK, Chan DJC
    Water Environ Res, 2021 Jan 23.
    PMID: 33484623 DOI: 10.1002/wer.1515
    Membrane distillation (MD) frequently deals with membrane biofouling caused by deposition of algal organic matter (AOM) from algal blooms, hampering the treatment efficiency. In this study, AOMs, which are soluble extracellular polymeric substance (sEPS), bounded EPS (bEPS), and internal organic matter (IOM) from three benthic species (Amphora coffeaeformis, Cylindrotheca fusiformis, and Navicula incerta) were exposed to a temperature range to resemble the MD process. Results showed that EPS had higher polysaccharide fraction than protein with 85.71%, 68.26%, and 71.91% for A. coffeaeformis, N. incerta, and C. fusiformis, respectively. Both the EPS polysaccharide and protein concentration linearly increase with temperature, but the opposite was true for IOM and high-molecular-weight (HMW) polysaccharide. At 80°C, 5812.94 μg/g out of 6304.28 μg/g polysaccharide in A. coffeaeformis was of low molecular weight (LMW); hence, these findings suggested that they were the major foulants to clog the narrow pores within virgin hydrophobic membrane, forming a conditioning layer followed by deposition of HMW and hydrophilic polysaccharides onto the macropores to cause irreversible fouling. Cell lysis occurring at higher temperature increases the total protein content about 25% within the EPS matrix, inducing membrane plugging via hydrophobic-hydrophobic interactions. Overall, the AOM composition at different temperatures will likely dictate the fouling severity in MD. PRACTITIONER POINTS: EPS production of three benthic diatoms was the highest at 80°C. EPS from diatoms consists of at least 75.29% of polysaccharides. Small molecular weight carbohydrates (<12 kDa) were potential foulants. Proteins of internal organic matter (>56%) give irreversible attachment towards membranes. A. coffeaeformis was considered as the most fouling diatoms with highest EPS amount of 6304.28 μg/g.
    Matched MeSH terms: Molecular Weight
  2. Ng IS, Ooi CW, Liu BL, Peng CT, Chiu CY, Chang YK
    Int J Biol Macromol, 2020 Jul 01;154:844-854.
    PMID: 32194127 DOI: 10.1016/j.ijbiomac.2020.03.127
    In this study, polyacrylonitrile (PAN) nanofiber membrane was prepared by an electrospinning technique. After alkaline hydrolysis, the ion-exchange nanofiber membrane (P-COOH) was grafted with chitosan molecules to form a chitosan-modified nanofiber membrane (P-COOH-CS). Poly(hexamethylene biguanide) (PHMB) was then covalently immobilized on P-COOH and P-COOH-CS to form P-COOH-PHMB and P-COOH-CS-PHMB, respectively. The nanofiber membranes were subjected to various surface analyses as well as to the evaluations of antibacterial activity against Escherichia coli. The optimal modification conditions for P-COOH-CS-PHMB were attained by water-soluble chitosan at 50 kDa of molecular weight, coupling pH at 7, and 0.05% (w/w) of PHMB. Within 10 min of treatment, the antibacterial rate was close to 100%. Under the similar conditions of antibacterial treatment, the P-COOH-CS-PHMB exhibited a better antibacterial efficacy than the P-COOH-PHMB. When the number of bacterial cells was increased by 2000 folds, both types of nanofiber membranes still maintained the antibacterial rate close to 100%. After five cycles of repeated antibacterial treatment, the antibacterial efficacy of P-COOH-PHMB was 96%, which was higher than that of P-COOH-CS-PHMB (83%). The experimental results revealed that the PHMB-modified nanofiber membranes can be suitably applied in water treatment such as water disinfection and biofouling control.
    Matched MeSH terms: Molecular Weight
  3. Nur Azira, T., Amin, I., Che Man, Y. B.
    MyJurnal
    Gelatin is widely used in food and pharmaceutical products. However, the addition of gelatin especially in food products becomes a controversial issue among Muslims due to its animal origin. Thus, the present study was aimed to detect and differentiate the origin of gelatin added in processed foods using a combination method of sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and Principal Component Analysis (PCA). Porcine gelatin had exhibited 11 prominent polypeptides compared to bovine gelatin with 2 prominent polypeptides. Polypeptides of both gelatin sources at molecular weight ranged from 53 to 220 kDa can be used to differentiate between porcine and bovine gelatins using PCA. The efficiency in extracting gelatin from processed foods by different solutions was also evaluated. Extraction of gelatin in processed foods by cold acetone and deionised water had exhibited a similar polypeptide patterns, suggesting both solutions are suitable. The study indicated that approach of a simple gelatin extraction combined with SDS-PAGE and PCA, may provide robust information for gelatin species differentiation of processed foods.
    Matched MeSH terms: Molecular Weight
  4. Usang MD, Ivanyuk FA, Ishizuka C, Chiba S
    Sci Rep, 2019 Feb 06;9(1):1525.
    PMID: 30728435 DOI: 10.1038/s41598-018-37993-7
    We have decomposed to symmetric and asymmetric modes the mass-TKE fission fragment distributions calculated by 4-dimensional Langevin approach and observed how the dominant fission mode and symmetric mode change as functions of [Formula: see text] of the fissioning system in the actinides and trans-actinide region. As a result, we found that the symmetric mode makes a sudden transition from super-long to super short fission mode around 254Es. The dominant fission modes on the other hand, are persistently asymmetric except for 258Fm, 259Fm and 260Md when the dominant fission mode suddenly becomes symmetric although it returns to the asymmetric mode around 256No. These correlated "twin transitions" have been known empirically by Darleane Hoffman and her group back in 1989, but for the first time we have given a clear explanation in terms of a dynamical model of nuclear fission. More specifically, since we kept the shape model parameters unchanged over the entire mass region, we conclude that the correlated twin transition emerge naturally from the dynamics in 4-D potential energy surface.
    Matched MeSH terms: Molecular Weight
  5. Ramesh, S., Shanti, R., Chin, S.F.
    ASM Science Journal, 2011;5(1):19-26.
    MyJurnal
    In this present study, a series of polymer electrolyte thin films were synthesized by incorporating different ratios of lithium triflate (LiCF3SO3) in a low molecular weight polyvinyl chloride (PVC) matrix by the solution casting technique. The incorporation of LiCF3SO3 suppressed the high degree of crystallinity in PVC enabling the system to possess an appreciable ionic conductivity. The ionic conductivity of the samples, with different LiCF3SO3 content, was determined by the aid of ac impedance spectroscopy. The highest ionic conductivity of 4.04  10–9 S cm–1 was identified for the composition of PVC: LiCF3SO3 (75:25). Further understanding of the ionic conductivity mechanism was based on temperature-dependent conductivity data which obeyed Arrhenius theory, indicating that the ionic conductivity enhancement was thermally assisted. The possible dipole-dipole interaction between the chemical constituents was confirmed with changes in cage peak, analysed using Fourier transform infrared spectroscopy.
    Matched MeSH terms: Molecular Weight
  6. Zheng W, Mutha NV, Heydari H, Dutta A, Siow CC, Jakubovics NS, et al.
    PeerJ, 2016;4:e1698.
    PMID: 27017950 DOI: 10.7717/peerj.1698
    Background. The gram-negative Neisseria is associated with two of the most potent human epidemic diseases: meningococcal meningitis and gonorrhoea. In both cases, disease is caused by bacteria colonizing human mucosal membrane surfaces. Overall, the genus shows great diversity and genetic variation mainly due to its ability to acquire and incorporate genetic material from a diverse range of sources through horizontal gene transfer. Although a number of databases exist for the Neisseria genomes, they are mostly focused on the pathogenic species. In this present study we present the freely available NeisseriaBase, a database dedicated to the genus Neisseria encompassing the complete and draft genomes of 15 pathogenic and commensal Neisseria species. Methods. The genomic data were retrieved from National Center for Biotechnology Information (NCBI) and annotated using the RAST server which were then stored into the MySQL database. The protein-coding genes were further analyzed to obtain information such as calculation of GC content (%), predicted hydrophobicity and molecular weight (Da) using in-house Perl scripts. The web application was developed following the secure four-tier web application architecture: (1) client workstation, (2) web server, (3) application server, and (4) database server. The web interface was constructed using PHP, JavaScript, jQuery, AJAX and CSS, utilizing the model-view-controller (MVC) framework. The in-house developed bioinformatics tools implemented in NeisseraBase were developed using Python, Perl, BioPerl and R languages. Results. Currently, NeisseriaBase houses 603,500 Coding Sequences (CDSs), 16,071 RNAs and 13,119 tRNA genes from 227 Neisseria genomes. The database is equipped with interactive web interfaces. Incorporation of the JBrowse genome browser in the database enables fast and smooth browsing of Neisseria genomes. NeisseriaBase includes the standard BLAST program to facilitate homology searching, and for Virulence Factor Database (VFDB) specific homology searches, the VFDB BLAST is also incorporated into the database. In addition, NeisseriaBase is equipped with in-house designed tools such as the Pairwise Genome Comparison tool (PGC) for comparative genomic analysis and the Pathogenomics Profiling Tool (PathoProT) for the comparative pathogenomics analysis of Neisseria strains. Discussion. This user-friendly database not only provides access to a host of genomic resources on Neisseria but also enables high-quality comparative genome analysis, which is crucial for the expanding scientific community interested in Neisseria research. This database is freely available at http://neisseria.um.edu.my.
    Matched MeSH terms: Molecular Weight
  7. Tommy YB, Lim TS, Noordin R, Saadatnia G, Choong YS
    BMC Struct Biol, 2012 Nov 27;12:30.
    PMID: 23181504 DOI: 10.1186/1472-6807-12-30
    BACKGROUND: Toxoplasma gondii is an intracellular coccidian parasite that causes toxoplasmosis. It was estimated that more than one third of the world population is infected by T. gondii, and the disease is critical in fetuses and immunosuppressed patients. Thus, early detection is crucial for disease diagnosis and therapy. However, the current available toxoplasmosis diagnostic tests vary in their accuracy and the better ones are costly.

    RESULTS: An earlier published work discovered a highly antigenic 12 kDa excretory-secretory (ES) protein of T. gondii which may potentially be used for the development of an antigen detection test for toxoplasmosis. However, the three-dimensional structure of the protein is unknown. Since epitope identification is important prior to designing of a specific antibody for an antigen-detection based diagnostic test, the structural elucidation of this protein is essential. In this study, we constructed a three dimensional model of the 12 kDa ES protein. The built structure possesses a thioredoxin backbone which consists of four α-helices flanking five β-strands at the center. Three potential epitopes (6-8 residues) which can be combined into one "single" epitope have been identified from the built structure as the most potential antibody binding site.

    CONCLUSION: Together with specific antibody design, this work could contribute towards future development of an antigen detection test for toxoplasmosis.

    Matched MeSH terms: Molecular Weight
  8. Leman, A.M., Fakhrurrazi Rahman, Dafit Feriyanto
    MyJurnal
    The simulation of the catalytic converter system is quite needed in order to characterize the catalyst and also
    optimizing the monolithic design for the gas emission in the catalytic converter and other related mechanism. The
    objective of this study is to obtain quantitative description of the gas emission in the catalytic converter system of
    natural powered automobile exhaust gas using ANSYS Software. This work will present a finite element calculation
    to predict and evaluate the mass transfer, energy balance and velocity of gas emission in the catalytic converter. The
    expected result for this research is to evaluate data of the gas emission obtained from the software to be compared with
    the manual experiment in order to verify the effectiveness of modified catalytic converter.
    Matched MeSH terms: Molecular Weight
  9. Shodiya, S., Azhar, A. A., Darus, A. N.
    MyJurnal
    HCFCs, in addition to destroying the ozone layer, have been recognized as a contributing factor that increases global warming. It is widely used as working fluid in window air-conditioning system, where capillary tube serves as an expansion device. Literature reports have shown that no single refrigerant can solve the problem of ozone layer depletion and global warming. Refrigerant HC290/HC600a/HFC407C mixture, an eco-friendly refrigerant, has been recognized as an alternative to HCFC22. The objective of this study is to, for cost effectiveness, develop an empirical correlation to predict the refrigerant HC290/ HC600a/HFC407C mixture mass flow rate using statistical experimental design approach. A review of relevant literature shows that refrigerant’s mass flow rate depends on condensing temperature, degree of subcooling, inner diameter and length of capillary tube. The relationship between the mass flow rate and the four independent variables was established as an empirical mathematical correlation using central composite design (CCD), a response surface methodology (RSM). This empirical correlation was examined using analysis of variance (ANOVA) of 5% level of significance. The results of these analysis showed that the correlation fitted well with the experimental data yielding an average and standard deviation of 1.05% and 2.62%, respectively. The validity of the present correlation was further assessed by comparing it with published empirical correlation in literature and the result showed that the present correlation is consistent.
    Matched MeSH terms: Molecular Weight
  10. Yin CY, Aroua MK, Daud WM
    Water Sci Technol, 2007;56(9):95-101.
    PMID: 18025736
    Palm shell activated carbon was modified via surface impregnation with polyethyleneimine (PEI) to enhance removal of Cu(2+) from aqueous solution in this study. The effect of PEI modification on batch adsorption of Cu(2+) as well as the equilibrium behavior of adsorption of metal ions on activated carbon were investigated. PEI modification clearly increased the Cu(2+) adsorption capacities by 68% and 75.86% for initial solution pH of 3 and 5 respectively. The adsorption data of Cu(2+) on both virgin and PEI-modified AC for both initial solution pH of 3 and 5 fitted the Langmuir and Redlich-Peterson isotherms considerably better than the Freundlich isotherm.
    Matched MeSH terms: Molecular Weight
  11. Abd Jalil SN, Wang DK, Yacou C, Motuzas J, Smart S, Diniz da Costa JC
    Materials (Basel), 2016 Nov 18;9(11).
    PMID: 28774057 DOI: 10.3390/ma9110938
    This work investigates the structural formation and analyses of titania membranes (TM) prepared using different vacuum exposure times for molecular weight (MW) cut-off performance and oil/water separation. Titania membranes were synthesized via a sol-gel method and coated on macroporous alumina tubes followed by exposure to a vacuum between 30 and 1200 s and then calcined at 400 °C. X-ray diffraction and nitrogen adsorption analyses showed that the crystallite size and particle size of titania increased as a function of vacuum time. All the TM membranes were mesoporous with an average pore diameter of ~3.6 nm with an anatase crystal morphology. Water, glucose, sucrose, and polyvinylpyrrolidone with 40 and 360 kDa (PVP-40 kDa and PVP-360 kDa) were used as feed solutions for MW cut-off and hexadecane solution for oil filtration investigation. The TM membranes were not able to separate glucose and sucrose, thus indicating the membrane pore sizes are larger than the kinetic diameter of sucrose of 0.9 nm, irrespective of vacuum exposure time. They also showed only moderate rejection (20%) of the smaller PVP-40 kDa, however, all the membranes were able to obtain an excellent rejection of near 100% for the larger PVP-360 kDa molecule. Furthermore, the TM membranes were tested for the separation of oil emulsions with a high concentration of oil (3000 ppm), reaching high oil rejections of more than 90% of oil. In general, the water fluxes increased with the vacuum exposure time indicating a pore structural tailoring effect. It is therefore proposed that a mechanism of pore size tailoring was formed by an interconnected network of Ti-O-Ti nanoparticles with inter-particle voids, which increased as TiO₂ nanoparticle size increased as a function of vacuum exposure time, and thus reduced the water transport resistance through the TM membranes.
    Matched MeSH terms: Molecular Weight
  12. Salleh, R.M., Djauhari, M.A.
    ASM Science Journal, 2012;6(1):1-13.
    MyJurnal
    A monitoring procedure was introduced for process variability in a multivariate setting based on individual observations which was a combination of (i) robust high breakdown point approach in the set-up stage to determine the reference sample and (ii) the use of Wilks chart in the mass production stage. This setting is what the Malaysian manufacturing industry is currently lacking in, especially when a robust approach must be used. The advantage of this procedure was revealed by using the case of a female shrouded connector production process in a Malaysian industry. Moreover, this procedure could also be used in any process quality monitoring and for any industry. A recommendation for quality practitioners was also addressed.
    Matched MeSH terms: Molecular Weight
  13. Sayyed RZ, Wani SJ, Alyousef AA, Alqasim A, Syed A, El-Enshasy HA
    PLoS One, 2019;14(6):e0212324.
    PMID: 31211775 DOI: 10.1371/journal.pone.0212324
    Poly-β-hydroxybutyrate (PHB) depolymerase is known to decompose PHB, biodegradable polymers and therefore has great commercial significance in the bioplastic sector. However, reports on PHB depolymerases from isolates obtained from plastic-contaminated sites that reflect the potential of the source organism is scarce. In this study, we evaluated the production of extracellular PHB depolymerase from Microbacterium paraoxydans RZS6 isolated from the plastic-contaminated site in the municipal area of Shahada, Maharashtra, India, for the first time. The isolate was identified using 16S rRNA gene sequencing, gas chromatographic analysis of fatty acid methyl esters (GC-FAME), and BIOLOG method. Ithydrolyzed PHB on minimal salt medium (MSM) containing PHB as the only source of carbon. The isolate produced PHB depolymerase at 45°C during 48 h of incubation. The enzyme was purified most efficiently using octyl-sepharose CL-4B column, with the highest purification yield of 6.675 Umg-1mL-1. The activity of the enzyme was enhanced in the presence of Ca2+ and Mg2+ ions but inhibited by Fe2+ (1 mM) ions and mercaptoethanol (1000 rpm). the nzyme kinetic analysis revealed that the enzyme was a metalloenzyme; requiring Mg2+ ions, that showed optimum enzyme activity at 30°C (mesophilic) and under neutrophilic (pH 7) conditions. Scale-up from the shake-flask level to a laboratory-scale bioreactor further enhanced the enzyme yield by 0.809 UmL-1. The molecular weight of the enzyme (40 kDa), as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, closely resembled the PHB depolymerase of Aureobacterium saperdae. Our findings highlighted the applicability of M. paraoxydans as a producer of extracellular PHB depolymerase having potential of degrading PHB under diverse conditions.
    Matched MeSH terms: Molecular Weight
  14. Sayyed RZ, Wani SJ, Alarfaj AA, Syed A, El-Enshasy HA
    PLoS One, 2020;15(1):e0220095.
    PMID: 31910206 DOI: 10.1371/journal.pone.0220095
    There are numerous reports on poly-β-hydroxybutyrate (PHB) depolymerases produced by various microorganisms isolated from various habitats, however, reports on PHB depolymerase production by an isolate from plastic rich sites scares. Although PHB has attracted commercial significance, the inefficient production and recovery methods, inefficient purification of PHB depolymerase and lack of ample knowledge on PHB degradation by PHB depolymerase have hampered its large scale commercialization. Therefore, to ensure the biodegradability of biopolymers, it becomes imperative to study the purification of the biodegrading enzyme system. We report the production, purification, and characterization of extracellular PHB depolymerase from Stenotrophomonas sp. RZS7 isolated from a dumping yard rich in plastic waste. The isolate produced extracellular PHB depolymerase in the mineral salt medium (MSM) at 30°C during 4 days of incubation under shaking. The enzyme was purified by three methods namely ammonium salt precipitation, column chromatography, and solvent purification. Among these purification methods, the enzyme was best purified by column chromatography on the Octyl-Sepharose CL-4B column giving optimum yield (0.7993 Umg-1mL-1). The molecular weight of purified PHB depolymerase was 40 kDa. Studies on the assessment of biodegradation of PHB in liquid culture medium and under natural soil conditions confirmed PHB biodegradation potential of Stenotrophomonas sp. RZS7. The results obtained in Fourier-Transform Infrared (FTIR) analysis, High-Performance Liquid Chromatography (HPLC) study and Gas Chromatography Mass-Spectrometry (GC-MS) analysis confirmed the biodegradation of PHB in liquid medium by Stenotrophomonas sp. RZS7. Changes in surface morphology of PHB film in soil burial as observed in Field Emission Scanning Electron Microscopy (FESEM) analysis confirmed the biodegradation of PHB under natural soil environment. The isolate was capable of degrading PHB and it resulted in 87.74% biodegradation. A higher rate of degradation under the natural soil condition is the result of the activity of soil microbes that complemented the biodegradation of PHB by Stenotrophomonas sp. RZS7.
    Matched MeSH terms: Molecular Weight
  15. Aidah Jumahat, Napisah Sapiai, Eliya Farah Hana Mohd Kamal
    MyJurnal
    This paper investigates the effect of acid and silane treatment of Carbon Nanotubes (CNT) on wear properties of epoxy polymer composite. The wear test done was based on ASTM D3389 standard using the Abrasive Wear Tester (TR 600). Characterisation analysis was also done using Transmission Electron Microscopy (TEM) in order to study the dispersion of the CNT inside the epoxy matrix. When untreated CNT was added to the epoxy with amounts of 0.5, 0.75 and 1.0 wt%, the wear rates did not improve except for 0.5 wt% CNT filled epoxy. This was due to the lack of dispersion which causes larger chunks of material being dug out, thus contributing to a higher mass loss and wear rate. When treated with acid and silane, 0.75 wt% and 1.0 wt% CNT filled epoxy composites showed improvement. The TEM images of 0.5 wt%, 0.75 wt% and 1.0 wt% PCNT filled epoxy supported the claim of the lack of dispersion of PCNT inside the epoxy.
    Matched MeSH terms: Molecular Weight
  16. Marmey P, Bothner B, Jacquot E, de Kochko A, Ong CA, Yot P, et al.
    Virology, 1999 Jan 20;253(2):319-26.
    PMID: 9918890
    Rice tungro bacilliform virus (RTBV) is a plant pararetrovirus and a member of the Caulimoviridae family and closely related to viruses in the Badnavirus genus. The coat protein of RTBV is part of the large polyprotein encoded by open reading frame 3 (ORF3). ORF3 of an RTBV isolate from Malaysia was sequenced (accession no. AF076470) and compared with published sequences for the region that encodes the coat protein or proteins. Molecular mass of virion proteins was determined by mass spectrometry (matrix-assisted laser desorption/ionization-TOF) performed on purified virus particles from three RTBV isolates from Malaysia. The N- and C-terminal amino acid sequences of the coat protein were deduced from the mass spectral analysis, leading to the conclusion that purified virions contain a single coat protein of 37 kDa. The location of the coat protein domain in ORF3 was reinforced as a result of immunodetection reactions using antibodies raised against six different segments of ORF3 using Western immunoblots after SDS-PAGE and isoelectrofocusing of proteins purified from RTBV particles. These studies demonstrate that RTBV coat protein is released from the polyprotein as a single coat protein of 37 kDa.
    Matched MeSH terms: Molecular Weight
  17. Shahid M, Azfaralariff A, Law D, Najm AA, Sanusi SA, Lim SJ, et al.
    Sci Rep, 2021 01 15;11(1):1594.
    PMID: 33452398 DOI: 10.1038/s41598-021-81026-9
    Xanthorrhizol (XNT), is a bioactive compound found in Curcuma xanthorrhiza Roxb. This study aimed to determine the potential targets of the XNT via computational target fishing method. This compound obeyed Lipinski's and Veber's rules where it has a molecular weight (MW) of 218.37 gmol-1, TPSA of 20.23, rotatable bonds (RBN) of 4, hydrogen acceptor and donor ability is 1 respectively. Besides, it also has half-life (HL) values 3.5 h, drug-likeness (DL) value of 0.07, oral bioavailability (OB) of 32.10, and blood-brain barrier permeability (BBB) value of 1.64 indicating its potential as therapeutic drug. Further, 20 potential targets were screened out through PharmMapper and DRAR-CPI servers. Co-expression results derived from GeneMANIA revealed that these targets made connection with a total of 40 genes and have 744 different links. Four genes which were RXRA, RBP4, HSD11B1 and AKR1C1 showed remarkable co-expression and predominantly involved in steroid metabolic process. Furthermore, among these 20 genes, 13 highly expressed genes associated with xenobiotics by cytochrome P450, chemical carcinogenesis and steroid metabolic pathways were identified through gene ontology (GO) and KEGG pathway analysis. In conclusion, XNT is targeting multiple proteins and pathways which may be exploited to shape a network that exerts systematic pharmacological effects.
    Matched MeSH terms: Molecular Weight
  18. Lau YL, Fong MY
    Exp Parasitol, 2008 Jul;119(3):373-8.
    PMID: 18457835 DOI: 10.1016/j.exppara.2008.03.016
    The full length surface antigen 2 (SAG2) gene of the protozoan parasite Toxoplasma gondii was cloned and intracellularly expressed in the Pichia pastoris expression system. The molecular weight of the expressed recombinant SAG2 (36 kDa) was much larger than the native SAG2 (22 kDa). This discrepancy in size was due to hyperglycosylation, as deglycosylation assay reduced the size of the recombinant SAG2 to 22 kDa. Despite being hyperglycosylated, the recombinant SAG2 reacted strongly with pooled anti-Toxoplasma human serum, pooled anti-Toxoplasma mouse serum and a SAG2-specific monoclonal antibody. The glycosylated recombinant SAG2 was further evaluated in Western blot and in-house enzyme-linked immunosorbent assay (ELISA) using 80 human serum samples, including confirmed early acute (IgM positive, IgG negative; n=20), acute (IgM positive, IgG positive; n=20) and chronic (IgM negative, IgG positive; n=20) toxoplasmosis patients, and toxoplasmosis negative control patients (n=20). Results of the Western blot showed that the recombinant SAG2 reacted with all 60 samples of the toxoplasmosis cases but not with the Toxoplasma-negative samples. The sensitivity of in-house ELISA was 80%, 95% and 100% for early acute, acute and chronic patients' serum samples, respectively. Vaccination study showed that serum from mice immunised with the glycosylated recombinant SAG2 reacted specifically with the native SAG2 of T. gondii. The mice were significantly protected against lethal challenge with live T. gondii RH strain tachyzoites (P<0.01) and their survival time was increased compared to controls. Therefore, the present study shows that the P. pastoris-derived recombinant SAG2 was specific and suitable for use as antigen for detecting anti-Toxoplasma IgG and IgM antibodies. The vaccination study showed that recombinant SAG2 protein was immunoprotective in mice against lethal challenge.
    Matched MeSH terms: Molecular Weight
  19. Pagthinathan, M., Ghazali, H.M., Yazid, A.M., Foo, H.L.
    MyJurnal
    Extracts from ‘kesinai’ (Streblus asper) leaves were investigated as a potential source of enzymes that can serve as an alternative to calf rennet in cheese making. Different types of extraction buffers were investigated namely sodium acetate buffer (pH 4.2-5.0), phosphate buffer (pH 6.0-7.0) and Tris-HCl buffer (pH 7.0-9.0). Finally, the milk-clotting enzyme was extracted using 100 mM Tris-HCl buffer (pH 7.4) with and without 5.0 mg/mL polyvinylpyrrolidone, 0.015 mL/mL Triton X-100 and 2 mM sodium metabisulphite. Purification was carried out using acetone precipitation, and ion-exchange and size-exclusion chromatographic techniques. Results showed that 100 mM Tris-HCl buffer (pH 7.4) was the most efficient extraction buffer among the buffers used in the extraction study. After the final purification step of size-exclusion chromatography, the enzyme was purified 3.3-fold with 42.3% of recovery. The enzyme showed an optimum temperature and pH at 60°C and pH 7.4, respectively. The enzyme was stable up to 70°C for one hour and the partially purified enzyme retained 83% and 96% of its original activity at pH 6.0 and 8.0, respectively. The molecular weight of the partially enzyme was estimated to be 75.8 kDa on SDS-PAGE. The milk-clotting activity of ‘kesinai’ enzyme was found to be lower than that of commercial Mucor rennet.
    Matched MeSH terms: Molecular Weight
  20. Kong BH, Teoh KH, Tan NH, Tan CS, Ng ST, Fung SY
    PeerJ, 2020;8:e9650.
    PMID: 32832273 DOI: 10.7717/peerj.9650
    Background: Lignosus tigris, a recently discovered species of the unique Lignosus family, has been traditionally used by the indigenous communities in Peninsular Malaysia to treat various ailments and as an alternative medicine for cancer treatment. The L. tigris cultivar sclerotia (Ligno TG-K) was found to contain numerous bioactive compounds with beneficial biomedicinal properties and the sclerotial extract exhibited potent antioxidant activity. However, the anticancer property of the Ligno TG-K including in vitro and in vivo antitumor effects as well as its anticancer active compounds and the mechanisms has yet to be investigated.

    Methods: The cytotoxicity of the Ligno TG-K against human breast (MCF7), prostate (PC3) and lung (A549) adenocarcinoma cell lines was evaluated using MTT cytotoxicity assay. The cytotoxic mechanisms of the active high molecular weight proteins (HMWp) fraction were investigated through detection of caspases activity and apoptotic-related proteins expression by Western blotting. The in vivo antitumor activity of the isolated HMWp was examined using MCF7 mouse xenograft model. Shotgun LC-MS/MS analysis was performed to identify the proteins in the HMWp.

    Results and Discussion: Cold water extract of the sclerotia inhibited proliferation of MCF7, A549 and PC3 cells with IC50 ranged from 28.9 to 95.0 µg/mL. Bioassay guided fractionation of the extract revealed that HMWp exhibited selective cytotoxicity against MCF7 cells via induction of cellular apoptosis by the activation of extrinsic and intrinsic signaling pathways. HMWp activated expression of caspase-8 and -9 enzymes, and pro-apoptotic Bax protein whilst inhibiting expression of tumor survivor protein, Bcl-2. HMWp induced tumor-cell apoptosis and suppressed growth of tumor in MCF-7 xenograft mice. Lectins, serine proteases, RNase Gf29 and a 230NA deoxyribonuclease are the major cytotoxic proteins that accounted for 55.93% of the HMWp.

    Conclusion: The findings from this study provided scientific evidences to support the traditional use of the L. tigris sclerotia for treatment of breast cancer. Several cytotoxic proteins with high abundance have been identified in the HMWp of the sclerotial extract and these proteins have potential to be developed into new anticancer agents or as adjunct cancer therapy.

    Matched MeSH terms: Molecular Weight
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