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  1. Loh KB, Ramli N, Tan LK, Roziah M, Rahmat K, Ariffin H
    Eur Radiol, 2012 Jul;22(7):1413-26.
    PMID: 22434420 DOI: 10.1007/s00330-012-2396-3
    OBJECTIVES: The degree and status of white matter myelination can be sensitively monitored using diffusion tensor imaging (DTI). This study looks at the measurement of fractional anistropy (FA) and mean diffusivity (MD) using an automated ROI with an existing DTI atlas.

    METHODS: Anatomical MRI and structural DTI were performed cross-sectionally on 26 normal children (newborn to 48 months old), using 1.5-T MRI. The automated processing pipeline was implemented to convert diffusion-weighted images into the NIfTI format. DTI-TK software was used to register the processed images to the ICBM DTI-81 atlas, while AFNI software was used for automated atlas-based volumes of interest (VOIs) and statistical value extraction.

    RESULTS: DTI exhibited consistent grey-white matter contrast. Triphasic temporal variation of the FA and MD values was noted, with FA increasing and MD decreasing rapidly early in the first 12 months. The second phase lasted 12-24 months during which the rate of FA and MD changes was reduced. After 24 months, the FA and MD values plateaued.

    CONCLUSION: DTI is a superior technique to conventional MR imaging in depicting WM maturation. The use of the automated processing pipeline provides a reliable environment for quantitative analysis of high-throughput DTI data.

    KEY POINTS: Diffusion tensor imaging outperforms conventional MRI in depicting white matter maturation. • DTI will become an important clinical tool for diagnosing paediatric neurological diseases. • DTI appears especially helpful for developmental abnormalities, tumours and white matter disease. • An automated processing pipeline assists quantitative analysis of high throughput DTI data.

    Matched MeSH terms: Image Enhancement/methods; Image Interpretation, Computer-Assisted/methods; Pattern Recognition, Automated/methods; Diffusion Magnetic Resonance Imaging/methods*
  2. Sachithanandan A
    Interact Cardiovasc Thorac Surg, 2011 Feb;12(2):120.
    PMID: 21257947 DOI: 10.1510/icvts.2010.252668A
    Matched MeSH terms: Reoperation/methods; Cardiovascular Surgical Procedures/methods; Negative-Pressure Wound Therapy/methods*; Sternotomy/methods
  3. Al-Odaini NA, Zakaria MP, Yaziz MI, Surif S
    J Chromatogr A, 2010 Oct 29;1217(44):6791-806.
    PMID: 20851398 DOI: 10.1016/j.chroma.2010.08.033
    Pollutants such as human pharmaceuticals and synthetic hormones that are not covered by environmental legislation have increasingly become important emerging aquatic contaminants. This paper reports the development of a sensitive and selective multi-residue method for simultaneous determination and quantification of 23 pharmaceuticals and synthetic hormones from different therapeutic classes in water samples. Target pharmaceuticals include anti-diabetic, antihypertensive, hypolipidemic agents, β2-adrenergic receptor agonist, antihistamine, analgesic and sex hormones. The developed method is based on solid phase extraction (SPE) followed by instrumental analysis using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) with 30 min total run time. River water samples (150 mL) and (sewage treatment plant) STP effluents (100 mL) adjusted to pH 2, were loaded into MCX (3 cm(3), 60 mg) cartridge and eluted with four different reagents for maximum recovery. Quantification was achieved by using eight isotopically labeled internal standards (I.S.) that effectively correct for losses during sample preparation and matrix effects during LC-ESI-MS/MS analysis. Good recoveries higher than 70% were obtained for most of target analytes in all matrices. Method detection limit (MDL) ranged from 0.2 to 281 ng/L. The developed method was applied to determine the levels of target analytes in various samples, including river water and STP effluents. Among the tested emerging pollutants, chlorothiazide was found at the highest level, with concentrations reaching up to 865 ng/L in STP effluent, and 182 ng/L in river water.
    Matched MeSH terms: Chromatography, Liquid/methods*; Spectrometry, Mass, Electrospray Ionization/methods; Solid Phase Extraction/methods*; Tandem Mass Spectrometry/methods*
  4. Norhaida A, Suharni M, Liza Sharmini AT, Tuda J, Rahmah N
    Ann Trop Med Parasitol, 2008 Mar;102(2):151-60.
    PMID: 18318937 DOI: 10.1179/136485908X252250
    Currently, the laboratory diagnosis of toxocariasis, caused by Toxocara canis or T. cati, mainly relies on serological tests. Unfortunately, however, the specificities of most of the commercial tests that are available for the serodiagnosis of this disease are not very high and this may cause problems, especially in tropical countries where co-infections with other helminths are common. In an effort to develop a serological assay with improved specificity for the detection of Toxocara infection, an IgG(4)-ELISA based on a recombinant version (rTES-30USM) of the 30-kDa Toxocara excretory-secretory antigen (TES-30) has recently been developed. To produce the antigen, the TES-30 gene was cloned via assembly PCR, subcloned into a His-tagged prokaryotic expression vector, and purified by affinity chromatography using Ni(2+)-nitrilotriacetic-acid (Ni-NTA) resin. The performance of the ELISA based on the recombinant antigen was then compared with that of commercial kit, based on an IgG-ELISA, for the serodiagnosis of toxocariasis (Toxocara IgG-ELISA; Cypress Diagnostics, Langdorp, Belgium). Both assays were used to test 338 serum samples, including 26 samples from probable cases of toxocariasis. Assuming that all the probable cases were true cases, the assay based on rTES-30USM demonstrated a sensitivity of 92.3% (24/26) and a specificity of 89.6% (103/115) whereas the commercial kit exhibited a sensitivity of 100% (26/26) but a specificity of only 55.7% (64/115). The high sensitivity and specificity exhibited by the new IgG(4)-ELISA should make the assay a good choice for use in tropical countries and any other area where potentially cross-reactive helminthic infections are common.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods*; Serologic Tests/methods; Polymerase Chain Reaction/methods; Targeted Gene Repair/methods
  5. Gopinath VK, Muda WA
    PMID: 15906679
    Feeding difficulties in cleft lip and palate (CLP) infants is commonly observed and is the most traumatic experience the family has to face. These infants are undernourished and have compromised growth. The purpose of this study was to 1) assess general health and growth parameters in children with CLP and in normal children; and 2) investigate the feeding methods of CLP infants and normal infants. A total of 221 children from birth to six years of both sexes, with CLP (60 children) and normal (161 children) were selected. The CLP and normal children were divided into three subgroups by age. The practice of feeding the infants in subgroup I was assessed using standard piloted questionnaires. The assessment of growth was done at baseline and at six months in all the subgroups.The general well being of the children was assessed by noting the number of common infections. Results showed that a significantly higher percentage of mothers with normal babies (p < 0.01) had a positive attitude towards breast feeding. When compared to normal children, CLP children were more susceptible to infections (p < 0.05) and measured significantly lower on the height growth curve(p < 0.05). Hence, height can be used to monitor growth in CLP children.
    Matched MeSH terms: Child Care/methods*; Feeding Methods*; Infant Care/methods*
  6. Kwan MK, Chiu CK, Lee CK, Chan CY
    Bone Joint J, 2015 Nov;97-B(11):1555-61.
    PMID: 26530660 DOI: 10.1302/0301-620X.97B11.35789
    Percutaneous placement of pedicle screws is a well-established technique, however, no studies have compared percutaneous and open placement of screws in the thoracic spine. The aim of this cadaveric study was to compare the accuracy and safety of these techniques at the thoracic spinal level. A total of 288 screws were inserted in 16 (eight cadavers, 144 screws in percutaneous and eight cadavers, 144 screws in open). Pedicle perforations and fractures were documented subsequent to wide laminectomy followed by skeletalisation of the vertebrae. The perforations were classified as grade 0: no perforation, grade 1: < 2 mm perforation, grade 2: 2 mm to 4 mm perforation and grade 3: > 4 mm perforation. In the percutaneous group, the perforation rate was 11.1% with 15 (10.4%) grade 1 and one (0.7%) grade 2 perforations. In the open group, the perforation rate was 8.3% (12 screws) and all were grade 1. This difference was not significant (p = 0.45). There were 19 (13.2%) pedicle fractures in the percutaneous group and 21 (14.6%) in the open group (p = 0.73). In summary, the safety of percutaneous fluoroscopy-guided pedicle screw placement in the thoracic spine between T4 and T12 is similar to that of the conventional open technique.
    Matched MeSH terms: Fluoroscopy/methods; Spinal Fusion/methods*; Radiography, Interventional/methods; Minimally Invasive Surgical Procedures/methods
  7. Mountjoy M, Akef N, Budgett R, Greinig S, Li G, Manikavasagam J, et al.
    Br J Sports Med, 2015 Jul;49(13):887-92.
    PMID: 25833900 DOI: 10.1136/bjsports-2014-094424
    Antidoping and medical care delivery programmes are required at all large international multisport events.
    Matched MeSH terms: Delivery of Health Care/methods*; Health Education/methods; Public Health Administration/methods; Substance Abuse Detection/methods*
  8. Tay ST, Rohani MY, Ho TM, Devi S
    Diagn Microbiol Infect Dis, 2002 Oct;44(2):137-42.
    PMID: 12458119
    In this study, recombinant proteins that encompassed the AD I-AD III regions of 56 kDa immunodominant gene of 2 Orientia tsutsugamushi (OT) serotypes; Gilliam and TA763 were expressed in Escherichia coli. Both recombinant proteins exhibited serologic cross-reactivity with the rabbit antisera against various OT serotypes, as evaluated by enzyme-linked immunosorbent assay (ELISA), but not against other rickettsial species, including Rickettsia typhi, R. prowazekii and TT118 SFG rickettsiae. The feasibility of using the recombinant proteins as a diagnostic reagent was further evaluated by ELISA using sera from blood donors and scrub typhus patients. The results suggested a higher affinity of the antihuman IgM than IgG with both recombinant proteins. The IgM ELISA findings were agreeable with the results of indirect immunoperoxidase (IIP) assay especially with sera of high antibody (1:1600). However, more than one antigen are probably needed for development of an effective assay for serodiagnosis of scrub typhus in endemic areas.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods*; Immunoenzyme Techniques/methods; Serologic Tests/methods; Polymerase Chain Reaction/methods*
  9. Zainal Z, Sajari R, Ismail I
    J. Biochem. Mol. Biol. Biophys., 2002 Dec;6(6):415-9.
    PMID: 14972797
    Ornithine decarboxylase (ODC) is an enzyme of one of the two pathways of putrescine biosynthesis in plants. The genes encoding ODC have previously been cloned from Datura stramonium and human. Using differential screening, we isolated ODC cDNA clone from a cDNA library of ripening Capsicum annuum fruit. The cDNA clone designated CUKM10 contains an insert of 1523 bp. The longest open reading frame potentially encodes a peptide of 345 amino acids with an estimated molecular mass of 47 kDa and exhibit striking similarity to other ODCs. Expression analysis showed that the capODC hybridised to a single transcript with a size of 1.7 kb. The capODC transcript was first observed in early ripening and increased steadily until it reached fully ripening stage. From the observation it is suggested that capODC is developmentally regulated especially during later stage of ripening.
    Matched MeSH terms: Cloning, Molecular/methods*; Sequence Analysis, DNA/methods*; Sequence Analysis, Protein/methods*; Gene Expression Profiling/methods*
  10. Phua AC, Abdullah RB, Mohamed Z
    J. Reprod. Dev., 2003 Aug;49(4):307-11.
    PMID: 14967923
    Sex determination of livestock is performed to achieve the objectives of livestock breeding programmes. Techniques for sex determination have evolved from karyotyping to detecting Y-specific antigens and recently to the polymerase chain reaction (PCR), which appears to be the most sensitive, accurate, rapid and reliable method to date. In this study, a PCR-based sex determination method for potential application in goat breeding programmes was developed. Primers were designed to amplify a portion of the X amelogenin gene (Aml-X) on the X chromosome to give a 300 bp product and Sry gene on the Y chromosome to give a 116 bp product. PCR optimization was performed using DNA template extracted from a whole blood sample of Jermasia goats (German Fawn x Katjang) of both sexes. It was possible to identify the sex chromosomes by amplifying both male- and female-specific genes simultaneously in a duplex reaction with males yielding two bands and females yielding one band. The Aml-X primer set, which served as an internal control primer, did not interfere with amplification of the Y-specific sequence even when a low amount of DNA (1 ng) was used. The duplex reaction subjected to a blind test showed 100% (14/14) concordance, proving its accuracy and reliability. The primer sets used were found to be highly specific and were suitable for gender selection of goats.
    Matched MeSH terms: Breeding/methods; Sex Determination Analysis/methods*; Sex Preselection/methods; Polymerase Chain Reaction/methods*
  11. Zhang ZY, Yang WY, Dominiczak AF, Wang JG, Wu Y, Almustafa B, et al.
    Hypertension, 2019 11;74(5):1064-1067.
    PMID: 31422692 DOI: 10.1161/HYPERTENSIONAHA.119.13206
    Matched MeSH terms: Blood Pressure Determination/methods; Monitoring, Physiologic/methods; Blood Pressure Monitoring, Ambulatory/methods*; Computed Tomography Angiography/methods
  12. Sunderasan E, Bahari A, Arif SA, Zainal Z, Hamilton RG, Yeang HY
    Clin Exp Allergy, 2005 Nov;35(11):1490-5.
    PMID: 16297147 DOI: 10.1111/j.1365-2222.2005.02371.x
    BACKGROUND:
    Hev b 4 is an allergenic natural rubber latex (NRL) protein complex that is reactive in skin prick tests and in vitro immunoassays. On SDS-polyacrylamide gel electrophoresis (SDS-PAGE), Hev b 4 is discerned predominantly at 53-55 kDa together with a 57 kDa minor component previously identified as a cyanogenic glucosidase. Of the 13 NRL allergens recognized by the International Union of Immunological Societies, the 53-55 kDa Hev b 4 major protein is the only candidate that lacks complete cDNA and protein sequence information.

    OBJECTIVE:
    We sought to clone the transcript encoding the Hev b 4 major protein, and characterize the native protein and its recombinant form in relation to IgE binding.

    METHODS:
    The 5'/3' rapid amplification of cDNA ends method was employed to obtain the complete cDNA of the Hev b 4 major protein. A recombinant form of the protein was over-expressed in Escherichia coli. The native Hev b 4 major protein was deglycosylated by trifluoromethane sulphonic acid. Western immunoblots of the native, deglycosylated and recombinant proteins were performed using both polyclonal antibodies and sera from latex-allergic patients.

    RESULTS:
    The cDNA encoding the Hev b 4 major protein was cloned. Its open reading frame matched lecithinases in the conserved domain database and contained 10 predicted glycosylation sites. Detection of glycans on the Hev b 4 lecithinase homologue confirmed it to be a glycoprotein. The deglycosylated lecithinase homologue was discerned at 40 kDa on SDS-PAGE, this being comparable to the 38.53 kDa mass predicted by its cDNA. Deglycosylation of the lecithinase homologue resulted in the loss of IgE recognition, although reactivity to polyclonal rabbit anti-Hev b 4 was retained. IgE from latex-allergic patients also failed to recognize the non-glycosylated E. coli recombinant lecithinase homologue.

    CONCLUSION:
    The IgE epitopes of the Hev b 4 lecithinase homologue reside mainly in its carbohydrate moiety, which also account for the discrepancy between the observed molecular weight of the protein and the value calculated from its cDNA.
    Matched MeSH terms: Cloning, Molecular/methods*; Electrophoresis, Polyacrylamide Gel/methods; Blotting, Western/methods; Reverse Transcriptase Polymerase Chain Reaction/methods
  13. Dahlan HM, Karsani SA, Rahman MA, Hamid NA, Top AG, Ngah WZ
    J Nutr Biochem, 2012 Jul;23(7):741-51.
    PMID: 21840697 DOI: 10.1016/j.jnutbio.2011.03.018
    Vitamin E has been suggested to modulate age-associated changes by altering the redox balance resulting in altered gene and/or protein expression. Here we have utilized proteomics to determine whether such regulation in protein expression occurs in human lymphocytes from two different age groups stressed with H₂O₂ and then treated with vitamin E in the form of tocotrienol-rich fraction (TRF). In this study, lymphocytes obtained from young (30-49 years old) and old (>50 years old) volunteers were first challenged with 1 mM H₂O₂. They were then treated by exposure to 50, 100 and 200 μg/ml TRF. Two-dimensional gel electrophoresis followed by MALDI-TOF/TOF (matrix-assisted laser desorption/ionization time-of-flight/time-of-flight) tandem mass spectrometry was then performed on whole-cell protein extracts to identify proteins that have changed in expression. A total of 24 proteins were found to be affected by H₂O₂ and/or TRF treatment. These included proteins that were related to metabolism, antioxidants, structural proteins, protein degradation and signal transduction. Of particular interest was the regulation of a number of proteins involved in stress response--peroxiredoxin-2, peroxiredoxin-3 and peroxiredoxin-6-all of which were shown to be down-regulated with H₂O₂ exposure. The effect was reversed following TRF treatment. The expression of peroxiredoxin-2 and peroxiredoxin-6 was confirmed by quantitative reverse transcriptase polymerase chain reaction. These results suggested that TRF directly influenced the expression dynamics of the peroxiredoxin-2, thus improving the cells ability to resist damage caused by oxidative stress.
    Matched MeSH terms: Electrophoresis, Gel, Two-Dimensional/methods; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods; Reverse Transcriptase Polymerase Chain Reaction/methods; Proteomics/methods*
  14. Poh AH, Adikan FRM, Moghavvemi M
    Med Biol Eng Comput, 2020 Jun;58(6):1159-1175.
    PMID: 32319030 DOI: 10.1007/s11517-019-02077-9
    The study and applications of in vivo skin optics have been openly documented as early as the year 1954, or possibly earlier. To date, challenges in analyzing the complexities of this field remain, with wide scopes requiring more scrutiny. Recent advances in spectroscopic research and multivariate analytics allow a closer look into applications potentially for detecting or monitoring diseases. One of the challenges in this field is in establishing a reference for applications which correspond to certain bandwidths. This article reviews the scope on past research on skin spectroscopy, and the clinical aspects which have or may have applications on disease detection or enhancing diagnostics. A summary is supplied on the technicalities surrounding the measurements reported in literature, focused towards the wavelength-dependent applications in themes central to the respective research. Analytics on the topology of the papers' data cited in this work is also provided for a statistical perspective. In short, this paper strives to immediately inform the reader with possible applications via the spectroscopic devices at hand. Graphical Abstract .
    Matched MeSH terms: Monitoring, Physiologic/methods; Oximetry/methods; Spectroscopy, Near-Infrared/methods; Optics and Photonics/methods*
  15. Daffalla SB, Mukhtar H, Shaharun MS
    PLoS One, 2020;15(12):e0243540.
    PMID: 33275643 DOI: 10.1371/journal.pone.0243540
    Rice husk is a base adsorbent for pollutant removal. It is a cost-effective material and a renewable resource. This study provides the physicochemical characterization of chemically and thermally treated rice husk adsorbents for phenol removal from aqueous solutions. We revealed new functional groups on rice husk adsorbents by Fourier transform infrared spectroscopy, and observed major changes in the pore structure (from macro-mesopores to micro-mesopores) of the developed rice husk adsorbents using scanning electron microscopy. Additionally, we studied their surface area and pore size distribution, and found a greater enhancement of the morphological structure of the thermally treated rice husk compared with that chemically treated. Thermally treated adsorbents presented a higher surface area (24-201 m2.g-1) than those chemically treated (3.2 m2.g-1). The thermal and chemical modifications of rice husk resulted in phenol removal efficiencies of 36%-64% and 28%, respectively. Thus, we recommend using thermally treated rice husk as a promising adsorbent for phenol removal from aqueous solutions.
    Matched MeSH terms: Microscopy, Electron, Scanning/methods; Spectroscopy, Fourier Transform Infrared/methods; Water Purification/methods*; Environmental Restoration and Remediation/methods
  16. Haranal M, Mood MC, Leong MC, Febrianti Z, Abdul Latiff H, Samion H, et al.
    Interact Cardiovasc Thorac Surg, 2020 08 01;31(2):221-227.
    PMID: 32437520 DOI: 10.1093/icvts/ivaa069
    OBJECTIVES: This study aims to review our institutional experience of ductal stenting (DS) on the growth of pulmonary arteries (PAs) and surgical outcomes of PA reconstruction in this subset of patients.

    METHODS: This is a retrospective study done in neonates and infants up to 3 months of age with duct-dependent pulmonary circulation who underwent DS from January 2014 to December 2015. Post-stenting PA growth, surgical outcomes of PA reconstruction, post-surgical re-interventions, morbidity and mortality were analysed.

    RESULTS: During the study period, 46 patients underwent successful DS, of whom 38 underwent presurgery catheterization and definite surgery. There was significant growth of PAs in these patients. Biventricular repair was done in 31 patients while 7 had univentricular palliation. Left PA augmentation was required in 13 patients, and 10 required central PA augmentation during surgery. The mean follow-up period post-surgery was 4.5 ± 1.5 years. No significant postoperative complications were seen. No early or follow-up post-surgery mortality was seen. Four patients required re-interventions in the form of left PA stenting based on the echocardiography or computed tomography evidence of significant stenosis.

    CONCLUSIONS: DS provides good short-term palliation and the growth of PAs. However, a significant number of stented patients require reparative procedure on PAs at the time of surgical intervention. Acquired changes in the PAs following DS may be the reason for reintervention following PA reconstruction.

    Matched MeSH terms: Cardiac Catheterization/methods*; Palliative Care/methods*; Vascular Surgical Procedures/methods*; Reconstructive Surgical Procedures/methods
  17. Tan XY, Misran A, Daim LDJ, Lau BYC
    Food Chem, 2021 May 01;343:128471.
    PMID: 33143964 DOI: 10.1016/j.foodchem.2020.128471
    Four different methods were evaluated to extract proteins from "Musang King" durian pulps and subsequently proteins with different abundance between fresh and long term frozen storage were identified using two-dimensional polyacrylamide gel electrophoresis coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometer analyses. The acetone-phenol method was found to produce good protein yields and gave the highest gel resolution and reproducibility. Differential protein analyses of the durian pulp revealed that 15 proteins were down-regulated and three other proteins were up-regulated after a year of frozen storage. Isoflavone reductase-like protein, S-adenosyl methionine synthase, and cysteine synthase isoform were up-regulated during frozen storage. The down-regulation of proteins in frozen durian pulps indicated that frozen storage has affected proteins in many ways, especially in their functions related to carbohydrate and energy metabolisms, cellular components, and transport processes. This study will enable future detailed investigations of proteins associated with quality attributes of durians to be studied.
    Matched MeSH terms: Chemical Fractionation/methods*; Electrophoresis, Gel, Two-Dimensional/methods*; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods*; Proteomics/methods
  18. Kow RY, Yuen JC, Ahmad Alwi AA, Abas MF, Low CL
    JBJS Case Connect, 2019 6 25;9(2):e0163.
    PMID: 31233428 DOI: 10.2106/JBJS.CC.18.00163
    CASE: A 17-year-old male sustained an open fracture of the right medial malleolus (MM) with significant bone and soft tissue loss following a motor-vehicle accident. Following serial wound debridement, his ankle was effectively reconstructed with MM antiglide plate stabilization, iliac autogenous bone graft, and a free radial forearm soft tissue flap.

    CONCLUSIONS: Open MM fracture with bone and soft tissue loss is rare. It is feasible to treat this injury with a novel surgical reconstruction technique involving autogenous bicortical iliac bone graft and radial forearm free flap.

    Matched MeSH terms: Debridement/methods; Radiography/methods; Bone Transplantation/methods; Reconstructive Surgical Procedures/methods
  19. He L, Mao Y, Zhang L, Wang H, Alias SA, Gao B, et al.
    BMC Biotechnol, 2017 02 28;17(1):22.
    PMID: 28245836 DOI: 10.1186/s12896-017-0343-8
    BACKGROUND: α-Amylase plays a pivotal role in a broad range of industrial processes. To meet increasing demands of biocatalytic tasks, considerable efforts have been made to isolate enzymes produced by extremophiles. However, the relevant data of α-amylases from cold-adapted fungi are still insufficient. In addition, bread quality presents a particular interest due to its high consummation. Thus developing amylases to improve textural properties could combine health benefits with good sensory properties. Furthermore, iron oxide nanoparticles provide an economical and convenient method for separation of biomacromolecules. In order to maximize the catalytic efficiency of α-amylase and support further applications, a comprehensive characterization of magnetic immobilization of α-amylase is crucial and needed.

    RESULTS: A novel α-amylase (AmyA1) containing an open reading frame of 1482 bp was cloned from Antarctic psychrotolerant fungus G. pannorum and then expressed in the newly constructed Aspergillus oryzae system. The purified recombinant AmyA1 was approximate 52 kDa. AmyA1 was optimally active at pH 5.0 and 40 °C, and retained over 20% of maximal activity at 0-20 °C. The K m and V max values toward soluble starch were 2.51 mg/mL and 8.24 × 10-2 mg/(mL min) respectively, with specific activity of 12.8 × 103 U/mg. AmyA1 presented broad substrate specificity, and the main hydrolysis products were glucose, maltose, and maltotetraose. The influence of AmyA1 on the quality of bread was further investigated. The application study shows a 26% increase in specific volume, 14.5% increase in cohesiveness and 14.1% decrease in gumminess in comparison with the control. AmyA1 was immobilized on magnetic nanoparticles and characterized. The immobilized enzyme showed improved thermostability and enhanced pH tolerance under neutral conditions. Also, magnetically immobilized AmyA1 can be easily recovered and reused for maximum utilization.

    CONCLUSIONS: A novel α-amylase (AmyA1) from Antarctic psychrotolerant fungus was cloned, heterologous expression in Aspergillus oryzae, and characterized. The detailed report of the enzymatic properties of AmyA1 gives new insights into fungal cold-adapted amylase. Application study showed potential value of AmyA1 in the food and starch fields. In addition, AmyA1 was immobilized on magnetic nanoparticles and characterized. The improved stability and longer service life of AmyA1 could potentially benefit industrial applications.

    Matched MeSH terms: Chemical Fractionation/methods; Protein Engineering/methods; Immunomagnetic Separation/methods; Food Industry/methods
  20. Mustafa MI, Al-Marzooq F, How SH, Kuan YC, Ng TH
    Trop Biomed, 2011 Dec;28(3):531-44.
    PMID: 22433882 MyJurnal
    Community-acquired pneumonia (CAP) is still a major cause of morbidity and mortality especially to children and compromised hosts, such as the old and those with underlying chronic diseases. Knowledge of pathogens causing CAP constitutes the basis for selection of antimicrobial treatment. Previous data have shown that etiological agents can be identified in only up to 50% of patients, but this figure can be improved by using polymerase chain reaction (PCR). This study was designed to evaluate multiplex real-time PCR as a method for rapid differential detection of five bacterial causes of CAP (Streptococcus pneumoniae, Burkholderia pseudomallei and atypical bacterial pathogens namely Mycoplasma pneumoniae, Chlamydophila pneumoniae and Legionella pneumophila) in CAP patients attending Hospital Tengku Ampuan Afzan (HTAA)/ Kuantan, Pahang, Malaysia. Two previously developed multiplex real-time PCR assays, duplex for the differential detection of S. pneumoniae and B. pseudomallei and triplex for the atypical bacterial pathogens, were used to detect a bacterial cause of CAP in blood and respiratory samples. Thus, 46 blood and 45 respiratory samples collected from 46 adult CAP patients admitted to HTAA were analysed by multiplex real-time PCR assays and conventional methods. The microbial etiology of CAP could be established for 39.1% (18/46) of CAP patients by conventional methods and this was increased to 65.2% (30/46) with the additional use of real-time PCR. The most frequently detected pathogens were S. pneumoniae (21.7% - all by PCR alone), Klebsiella pneumoniae (17.3%), B. pseudomallei (13% - 83% of them positive by PCR alone and 17% by both culture and PCR), Pseudomonas aeruginosa (6.5%), M. pneumoniae (6.5% - all by serology), C. pneumoniae (4.3% - all positive by both PCR and serology), L. pneumophila (2.1% - all by PCR alone), Escherichia coli (4.3%). Haemophilus infuenzae, Acinetobacter lwoffii and Acinetobacter baumannii were detected by conventional methods (2.1% for each).
    Matched MeSH terms: Bacteriological Techniques/methods*; Molecular Diagnostic Techniques/methods*; Multiplex Polymerase Chain Reaction/methods*; Real-Time Polymerase Chain Reaction/methods*
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