Displaying publications 41 - 60 of 314 in total

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  1. Rosmilah M, Shahnaz M, Patel G, Lock J, Rahman D, Masita A, et al.
    Trop Biomed, 2008 Dec;25(3):243-51.
    PMID: 19287364 MyJurnal
    Royal jelly is widely consumed in the community and has perceived benefits ranging from promoting growth in children and improvement of general health status to enhancement of longevity for the elderly. However, royal jelly consumption has been linked to contact dermatitis, acute asthma, anaphylaxis and death. High prevalence of positive skin tests to royal jelly have been reported among atopic populations in countries with a high rate of royal jelly consumption. The present study is aimed to identify the major allergens of royal jelly. Royal jelly extract was separated by sodium dodecyl polyacrylamide gel electrophoresis (SDS-PAGE) and 2-dimensional electrophoresis (2-D). Immunoblotting of the SDS-PAGE and 2-D profiles were performed to identify the allergenic spots. Spots were then excised from the 2-D gel, digested with trypsin and analyzed by mass spectrometry. The SDS-PAGE of royal jelly extract revealed 18 bands between 10 to 167 kD. Western blot of the fractionated proteins detected 15 IgE-binding bands between 14 to 127 kD with seven major allergens of 32, 40, 42, 49, 55, 60 and 67 kD using serum from 53 subjects with royal jelly allergy. The 2-D gel fractionated the royal jelly proteins to more than 50 different protein spots. Out of these, 30 spots demonstrated specific IgE affinity to the sera tested. Eight spots of the major royal jelly allergens were selected for mass-spectrometry analysis. Digested tryptic peptides of the spots were compared to the amino acid sequence search in protein databases which identified the fragments of royal jelly homologus to major royal jelly protein 1 (MRJ1) and major royal jelly protein 2 (MRJ2). In conclusion, the major allergens of royal jelly are MRJ1 and MRJ2 in our patients' population.
    Matched MeSH terms: Molecular Weight
  2. Saad B, Wai WT, Lim BP
    J Oleo Sci, 2008;57(4):257-61.
    PMID: 18332590
    A comparative study of oxidative decomposition behavior of a wide range of vegetable oils and its correlation to iodine value (IV) using thermogravimetric analysis (TGA) was described. The oxidative decomposition of saturated fatty acids shows weight loss before 385 degrees C while oxidative decomposition of unsaturated fatty acids shows lower rate of weight loss (dWt/dt) compared to saturated fatty acids due to the oxidation process ('up taking ' of oxygen) involving breaking down of double bond to form primary and secondary oxidation products, which leads to some weight gain in the sample before being decomposed. The relative differences in the dWt/dt (%/min) of the both fatty acids give different decomposition steps in TGA thermogram, enabling IV to be determined through the percentage weight loss of saturated fatty acids per 100% of total sample weight (excluding weight loss from moisture and volatile compounds). Therefore, TGA method can be used as an alternative method for IV determination with no sample pre-dilution and solvent consumption. Using the TGA methods, good correlation (r = 0.9889) with standard AOCS method was achieved.
    Matched MeSH terms: Molecular Weight
  3. Noraida Mohamed Shah, Azmi Sarriff, Rosnani Hashim
    MyJurnal
    Low-molecular-weight heparins (LMWHs) are antithrombotic agents utilised in the treatment of acute coronary syndromes. They have been shown to be more effective than unfractionated heparins (UFHs) in reducing ischeamic events, which include death, myocardial infarction (MI) and urgent revascularisation. Enoxaparin is one of the products of LMWHs. Its safety and efficacy has been proven in the ESSENCE and TIMI IIB studies. This study was carried out to identify risk factors that may affect bleeding complications associated with the use of enoxaparin for non-ST-elevation MI (NSTEMI) or unstable angina (UA) in Universiti Kebangsaan Malaysia Hospital (HUKM). This observational, longitudinal study was conducted on patients who were admitted to the Coronary Care Unit (CCU), Coronary Rehabilitation Ward (CRW), Medical 1 and Medical 2 wards at HUKM and initiated on enoxaparin for NSTEMI/UA from 22nd of March until 22nd of April 2004. A total of 40 patients were included in the study with median age of 65 years, male to female ratio of 3:1, diagnosed with NSTEMI (55%) and UA (45%). 45% of patients developed an episode of bleeding and among them 83.3% (15 patients) characterised by haematuria. Higher percentages of women (80%) and those with creatinine clearance of < 30ml/min (100%) had incidence of bleeding as compared to men (50%) and those with creatinine clearance = 30 ml/min, respectively (p < 0.05 for both parameters). Age, enoxaparin dose and duration of therapy, smoking and concomitant aspirin/ticlopidine therapy did not significantly affect the incidence of bleeding. In conclusion, renal impairment and gender were associated with bleeding in relation with the use of enoxaparin that may require dose adjustments.
    Matched MeSH terms: Heparin, Low-Molecular-Weight
  4. Ravi S, Peh KK, Darwis Y, Murthy BK, Singh TR, Mallikarjun C
    Indian J Pharm Sci, 2008 May-Jun;70(3):303-9.
    PMID: 20046737 DOI: 10.4103/0250-474X.42978
    The aim of the present work was to investigate the preparation of microspheres as potential drug carriers for proteins, intended for controlled release formulation. The hydrophilic bovine serum albumin was chosen as a model protein to be encapsulated within poly(D,L-lactide-co-glycolide) (50:50) microspheres using a w/o/w double emulsion solvent evaporation method. Different parameters influencing the particle size, entrapment efficiency and in vitro release profiles were investigated. The microspheres prepared with different molecular weight and hydrophilicity of poly(D,L-lactide-co-glycolide) polymers were non porous, smooth surfaced and spherical in structure under scanning electron microscope with a mean particle size ranging from 3.98 to 8.74 mum. The protein loading efficiency varied from 40 to 71% of the theoretical amount incorporated. The in vitro release profile of bovine serum albumin from microspheres presented two phases, initial burst release phase due to the protein adsorbed on the microsphere surface, followed by slower and continuous release phase corresponding to the protein entrapped in polymer matrix. The release rate was fairly constant after an initial burst release. Consequently, these microspheres can be proposed as new controlled release protein delivery system.
    Matched MeSH terms: Molecular Weight
  5. Vigneswari S, Vijaya S, Majid MI, Sudesh K, Sipaut CS, Azizan MN, et al.
    J Ind Microbiol Biotechnol, 2009 Apr;36(4):547-56.
    PMID: 19189144 DOI: 10.1007/s10295-009-0525-z
    Cupriavidus sp. USMAA1020, a local isolate was able to biosynthesis poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] copolymer with various 4HB precursors as the sole carbon source. Manipulation of the culture conditions such as cell concentration, phosphate ratio and culture aeration significantly affected the synthesis of P(3HB-co-4HB) copolymer and 4HB composition. P(3HB-co-4HB) copolymer with 4HB compositions ranging from 23 to 75 mol% 4HB with various mechanical and thermal properties were successfully produced by varying the medium aeration. The physical and mechanical properties of P(3HB-co-4HB) copolymers were characterized by NMR spectroscopy, gel-permeation chromatography, tensile test, and differential scanning calorimetry. The number-average molecular weights (M (n)) of copolymers ranged from 260 x 10(3) to 590 x 10(3)Da, and the polydispersities (M (w)/M (n)) were between 1.8 and 3.0. Increases in the 4HB composition lowered the molecular weight of these copolymers. In addition, the increase in 4HB composition affected the randomness of copolymer, melting temperature (T (m)), glass transition temperature (T (g)), tensile strength, and elongation to break. Enzymatic degradation of P(3HB-co-4HB) films with an extracellular depolymerase from Ochrobactrum sp. DP5 showed that the degradation rate increased proportionally with time as the 4HB fraction increased from 17 to 50 mol% but were much lower with higher 4HB fraction. Degradation of P(3HB-co-4HB) films with lipase from Chromobacterium viscosum exhibited highest degradation rate at 75 mol% 4HB. The biocompatibility of P(3HB-co-4HB) copolymers were evaluated and these copolymers have been shown to support the growth and proliferation of fibroblast cells.
    Matched MeSH terms: Molecular Weight
  6. Kuan YH, Bhat R, Senan C, Williams PA, Karim AA
    J Agric Food Chem, 2009 Oct 14;57(19):9154-9.
    PMID: 19757813 DOI: 10.1021/jf9015625
    The impact of ultraviolet (UV) irradiation on the physicochemical and functional properties of gum arabic was investigated. Gum arabic samples were exposed to UV irradiation for 30, 60, 90, and 120 min; gum arabic was also treated with formaldehyde for comparison. Molecular weight analysis using gel permeation chromatography indicated that no significant changes occurred on the molecular structure on the samples exposed to UV irradiation. Free amino group analysis indicated that mild UV irradiation (30 min) could induce cross-linking on gum arabic; this result was comparable with that of samples treated with formaldehyde. However, viscosity break down was observed for samples exposed to UV irradiation for longer times (90 and 120 min). All irradiated and formaldehyde-treated samples exhibited better emulsification properties than unirradiated samples. These results indicate that UV-irradiated gum arabic could be a better emulsifier than the native (unmodified) gum arabic and could be exploited commercially.
    Matched MeSH terms: Molecular Weight
  7. Siew EL, Rajab NF, Osman AB, Sudesh K, Inayat-Hussain SH
    J Biomed Mater Res A, 2009 Dec;91(3):786-94.
    PMID: 19051306 DOI: 10.1002/jbm.a.32290
    Polyhydroxyalkanoates (PHA) are naturally occurring biopolyesters that have great potential in the medical field. However, the leachables resulting from sterilization process of the biomaterials may exert toxic effect including genetic damage. Here, we demonstrate that although gamma-irradiation of poly(3-hydroxybutyrate-co-50 mol % 4-hydroxybutyrate) [P(3HB-co-4HB)] did not cause any change in the morphology by scanning electron microscopy, there was a significant degradation of this copolymer where the molecular weight was reduced by 37% after sterilization indicating the generation of leachables. Therefore, further investigation on the ability of the extract of this poststerilized copolymer to induce mutagenic effect was performed using Ames test (S. typhimurium strains TA1535 and TA1537) and umu test (S. typhimurium strain TA1535/pSK1002). Additionally, the capability of the extract to induce clastogenic effect was determined using Chinese hamster lung V79 fibroblast cells. Our results showed that with and without the presence of S9 metabolic activation, no mutagenic effects were observed in both Ames and umu tests when treated with P(3HB-co-4HB) extract. Similarly, treatment of P(3HB-co-4HB) extract in V79 fibroblast cells showed no significant production of micronuclei when compared with the positive control (Mitomycin C). Together, these results indicate that leachables of poststerilized P(3HB-co-4HB) cause no mutagenic and clastogenic effects.
    Matched MeSH terms: Molecular Weight
  8. Arsad H, Sudesh K, Nazalan N, Muhammad TS, Wahab H, Razip Samian M
    Trop Life Sci Res, 2009 Dec;20(2):1-14.
    PMID: 24575175 MyJurnal
    The (R)-3-hydroxyacyl-ACP-CoA transferase catalyses the conversion of (R)-3-hydroxyacyl-ACP to (R)-3-hydroxyacyl-CoA derivatives, which serves as the ultimate precursor for polyhydroxyalkanoate (PHA) polymerisation from unrelated substrates in pseudomonads. PhaG was found to be responsible for channelling precursors for polyhydroxyalkanoate (PHA) synthase from a de novo fatty acid biosynthesis pathway when cultured on carbohydrates, such as glucose or gluconate. The phaG gene was cloned from Pseudomonas sp. USM 4-55 using a homologous probe. The gene was located in a 3660 bp Sal I fragment (GenBank accession number EU305558). The open reading frame (ORF) was 885 bp long and encoded a 295 amino acid protein. The predicted molecular weight was 33251 Da, and it showed a 62% identity to the PhaG of Pseudomonas aeruginosa. The function of the cloned phaG of Pseudomonas sp. USM 4-55 was confirmed by complementation studies. Plasmid pBCS39, which harboured the 3660 bp Sal I fragment, was found to complement the PhaG-mutant heterologous host cell, Pseudomonas putida PhaGN-21. P. putida PhaGN-21, which harboured pBCS39, accumulated PHA that accounted for up to 18% of its cellular dry weight (CDW). P. putida PhaGN-21, which harboured the vector alone (PBBR1MCS-2), accumulated only 0.6% CDW of PHA.
    Matched MeSH terms: Molecular Weight
  9. Yakubu R, Peh KK, Tan YT
    Drug Dev Ind Pharm, 2009 Dec;35(12):1430-8.
    PMID: 19929202 DOI: 10.3109/03639040902988566
    The purpose of this study was to design a 24-hour controlled porosity osmotic pump system that utilizes polyvinyl pyrrolidone (PVP) as an osmotic-suspending/release retarding agent of drugs.
    Matched MeSH terms: Molecular Weight
  10. Bakhshan, Y., Shahrir Abdullah
    MyJurnal
    An in-house quasi-dimensional code has been developed which simulate the overlap, intake, compression, combustion, as well as expansion and exhaust processes of a homogeneous charged internal combustion engine (ICE). A detailed chemical kinetic mechanism, constituting of 39 species and 148 elementary reactions, has been used in conjunction with above code to study the combustion of CNG under IC engine conditions. Two different criteria, based on pressure rise and mass of fuel burned, are used to detect the onset of ignition. Parametric studies are conducted to show the effect of compression ratio, initial pressure, intake temperature and equivalence ratio, on the time of ignition and fuel burning rate. The results obtained from the modelling show a good agreement with the experimental data.
    Matched MeSH terms: Molecular Weight
  11. Muskhazli Mustafa, Nor Azwady Abd. Aziz, Anida Kaimi, Nurul Shafiza Noor, Salifah Hasanah Ahmad Bedawi, Nalisha Ithnin
    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: Molecular Weight
  12. Tan NH
    PMID: 19770070 DOI: 10.1016/j.cbpc.2009.09.002
    A thrombin-like enzyme, purpurase, was purified from the Cryptelytrops purpureomaculatus (mangrove pit viper) venom using high performance ion-exchange and gel filtration chromatography. The purified sample (termed purpurase) yielded a homogeneous band in SDS-polyacrylamide gel electrophoresis with a molecular weight of 35,000. The N-terminal sequence of purpurase was determined to be VVGGDECNINDHRSLVRIF and is homologous to many other venom thrombin-like enzymes. Purpurase exhibits both arginine ester hydrolase and amidase activities. Kinetic studies using tripeptide chromogenic anilide substrates showed that purpurase is not fastidious towards its substrate. The clotting times of fibrinogen by purpurase were concentration dependent, with optimum clotting activity at 3mg fibronogen/mL. The clotting activity by purpurase was in the following decreasing order: cat fibrinogen>human fibrinogen>dog fibrinogen>goat fibrinogen>rabbit fibrinogen. Reversed-phase HPLC analysis of the products of action of purpurase on bovine fibrinogen showed that only fibrinopeptide A was released. Indirect ELISA studies showed that anti-purpurase cross-reacted strongly with venoms of most crotalid venoms, indicating the snake venom thrombin-like enzymes generally possess similar epitopes. In the more specific double-sandwich ELISA, however, anti-purpurase cross-reacted only with venoms of certain species of the Trimeresurus complex, and the results support the recent proposed taxonomy changes concerning the Trimeresurus complex.
    Matched MeSH terms: Molecular Weight
  13. Abdullah AZ, Ling PY
    J Hazard Mater, 2010 Jan 15;173(1-3):159-67.
    PMID: 19740600 DOI: 10.1016/j.jhazmat.2009.08.060
    The ambient sonocatalytic degradation of congo red, methyl orange, and methylene blue by titanium dioxide (TiO(2)) catalyst at initial concentrations between 10 and 50mg/L, catalyst loadings between 1.0 and 3.0mg/L and hydrogen peroxide (H(2)O(2)) concentrations up to 600 mg/L is reported. A 20 kHz ultrasonic processor at 50 W was used to accelerate the reaction. The catalysts were exposed to heat treatments between 400 and 1000 degrees C for up to 4h to induce phase change. Sonocatalysts with small amount of rutile phase showed better sonocatalytic activity but excessive rutile phase should be avoided. TiO(2) heated to 800 degrees C for 2h showed the highest sonocatalytic activity and the degradation of dyes was influenced by their chemical structures, chemical phases and characteristics of the catalysts. Congo red exhibited the highest degradation rate, attributed to multiple labile azo bonds to cause highest reactivity with the free radicals generated. An initial concentration of 10mg/L, 1.5 g/L of catalyst loading and 450 ppm of H(2)O(2) gave the best congo red removal efficiency of above 80% in 180 min. Rate coefficients for the sonocatalytic process was successfully established and the reused catalyst showed an activity drop by merely 10%.
    Matched MeSH terms: Molecular Weight
  14. Ho YC, Norli I, Alkarkhi AF, Morad N
    Bioresour Technol, 2010 Feb;101(4):1166-74.
    PMID: 19854044 DOI: 10.1016/j.biortech.2009.09.064
    Polyacrylamide (PAM), a commonly used organic synthetic flocculant, is known to have high reduction in turbidity treatment. However, PAM is not readily degradable. In this paper, pectin as a biopolymeric flocculant is used. The objectives are (i) to determine the characteristics of both flocculants (ii) to optimize the treatment processes of both flocculants in synthetic turbid waste water. The results obtained indicated that pectin has a lower average molecular weight at 1.63 x 10(5) and PAM at 6.00 x 10(7). However, the thermal degradation results showed that the onset temperature for pectin is at 165.58 degrees C, while the highest onset temperature obtained for PAM is at 235.39 degrees C. The optimum treatment conditions for the biopolymeric flocculant for flocculating activity was at pH 3, cation concentration at 0.55 mM, and pectin concentration at 3 mg/L. In contrast, PAM was at pH 4, cation concentration >0.05 mM and PAM concentration between 13 and 30 mg/L.
    Matched MeSH terms: Molecular Weight
  15. Che Yaakob CA, Dzarr AA, Ismail AA, Zuky Nik Lah NA, Ho JJ
    PMID: 20556784 DOI: 10.1002/14651858.CD007801.pub2
    BACKGROUND: Thromboembolic complications are much higher in pregnancy due to procoagulant changes. Heparin does not cross the placenta and the use of unfractionated heparin (UFH) is the current established practice in prophylaxis and treatment for thromboembolism in pregnancy.

    OBJECTIVES: To compare the effectiveness of anticoagulant therapies for the treatment of deep vein thrombosis in pregnancy. The anticoagulant drugs included are UFH, low molecular weight heparin (LMWH) and warfarin.

    SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group's Trials Register (March 2010) and reference lists of retrieved studies.

    SELECTION CRITERIA: Randomised controlled trials comparing any combination of warfarin, UFH, LMWH and placebo in pregnant women.

    DATA COLLECTION AND ANALYSIS: We used methods described in the Cochrane Handbooks for Systemic Reviews of Interventions for assessing the eligibility of studies identified by the search strategy. A minimum of two review authors independently assessed each study.

    MAIN RESULTS: We did not identify any eligible studies for inclusion in the review.We identified three potential studies; after assessing eligibility, we excluded all three as they did not meet the prespecified inclusion criteria. One study compared LMWH and UFH in pregnant women with previous thromboembolic events and, for most of these women, anticoagulants were used as thromboprophylaxis. There were only three women who had a thromboembolic event during the current pregnancy and it was unclear whether the anticoagulant was used as therapy or prophylaxis. We excluded one study because it included only women undergoing caesarean birth. The third study was not a randomised trial.

    AUTHORS' CONCLUSIONS: There is no evidence from randomised controlled trials on the effectiveness of anticoagulation for deep vein thrombosis in pregnancy. Further studies are required.

    Matched MeSH terms: Heparin, Low-Molecular-Weight/therapeutic use
  16. Owlad M, Aroua MK, Wan Daud WM
    Bioresour Technol, 2010 Jul;101(14):5098-103.
    PMID: 20156679 DOI: 10.1016/j.biortech.2010.01.135
    Removal of Cr(VI) ions from aqueous solution was investigated using modified palm shell activated carbon. Low Molecular Weight Polyethyleneimine (LMW PEI) was used for impregnation purpose. The maximum amount of LMW PEI adsorbed on activated carbon was determined to be approximately 228.2mg/g carbon. The adsorption experiments were carried out in a batch system using potassium dichromate K(2)Cr(2)O(7) as the source of Cr(VI) in the synthetic waste water and modified palm shell activated carbon as the adsorbent. The effects of pH, concentration of Cr(VI) and PEI loaded on activated carbon were studied. The adsorption data were found to fit well with the Freundlich isotherm model. This modified Palm shell activated carbon showed high adsorption capacity for chromium ions.
    Matched MeSH terms: Molecular Weight
  17. Ibrahim MZ, Norashikin MZ
    J Nanosci Nanotechnol, 2010 Sep;10(9):6211-5.
    PMID: 21133176
    This paper reports the performance of two different artificial neural networks (ANN), Multi Layer Perceptron (MLP) and Radial Basis Function (RBF) compared to conventional software for prediction of the pore size of the asymmetric polyethersulfone (PES) ultrafiltration membranes. ANN has advantages such as incredible approximation, generalization and good learning ability. The MLP are well suited for multiple inputs and multiple outputs while RBF are powerful techniques for interpolation in multidimensional space. Three experimental data sets were used to train the ANN using polyethylene glycol (PEG) of different molecular weights as additives namely as PEG 200, PEG 400 and PEG 600. The values of the pore size can be determined manually from the graph and solve it using mathematical equation. However, the mathematical solution used to determine the pore size and pore size distribution involve complicated equations and tedious. Thus, in this study, MLP and RBF are applied as an alternative method to estimate the pore size of polyethersulfone (PES) ultrafiltration membranes. The raw data needed for the training are solute separation and solute diameter. Values of solute separation were obtained from the ultrafiltration experiments and solute diameters ware calculated using mathematical equation. With the development of this ANN model, the process to estimate membrane pore size could be made easier and faster compared to mathematical solutions.
    Matched MeSH terms: Molecular Weight
  18. Arifin N, Basuni M, Lan CA, Yahya AR, Noordin R
    Protein J, 2010 Oct;29(7):509-15.
    PMID: 20845068 DOI: 10.1007/s10930-010-9281-1
    This paper describes a refinement in the purification step that facilitated the downstream recovery of high purity BmR1 recombinant protein, which is a protein used as a test reagent in the commercialized rapid tests for detection of lymphac filariasis i.e. Brugia Rapid™ and panLF rapid™. Purification was performed by immobilized metal affinity chromatography (IMAC), followed by ion exchange chromatography (IEX). Results showed that a total of 10.27 mg of BmR1 was obtained when IMAC was performed using 20 mM of imidazole and 5 column volume of wash buffer containing 500 mM of NaCl. Purity of the target protein was enhanced when buffer at pH 5.8 was used during the IEX. Two proteins that recurrently appeared below the BmR1 recombinant protein were identified by mass-spectrometry analysis as the same protein, thus they were probably degradation products of BmR1. These strategies improve purity of the target protein to be used in applications such as production of aptamers and monoclonal antibodies.
    Matched MeSH terms: Molecular Weight
  19. Lau YL, Hasan MT, Thiruvengadam G, Idris MM, Init I
    Trop Biomed, 2010 Dec;27(3):525-33.
    PMID: 21399595
    GRA4 of Toxoplasma gondii has been shown to prompt IgG, IgM and IgA responses in previous studies and is thus considered one of the major immunogenic proteins from T. gondii that can be used for both diagnostics purposes and vaccine development. This study seeks to clone and express the GRA4 in Pichia pastoris, which has numerous advantages over other systems for expression of eukaryotic proteins. In order to achieve this, the gene was cloned into the pPICZα A expression vector, which was then incorporated into the P. pastoris genome via insertional integration for expression of the recombinant protein, under the AOX1 promoter. The antigen was expressed along with the prepro sequence of the α-factor of yeast so that it could be excreted out of the P. pastoris cells and obtained from the medium. Upon SDS-PAGE analysis it was found that the recombinant protein was expressed optimally as a 40 kDa protein after 96 hours of induction with 0.75% of methanol. The expressed GRA4 protein showed discrepancy in size with the calculated molecular mass. This may be attributed to the various posttranslational modifications including glycosylation and phosphorylation. Despite the difference in molecular weight, the recombinant protein was able to detect toxoplasmosis in Western blot format. The recombinant GRA4 was expressed with an intact polyhistidine-tag, which could be used for future purification of the antigen.
    Matched MeSH terms: Molecular Weight
  20. Tan HY, Nagoor NH, Sekaran SD
    Trop Biomed, 2010 Dec;27(3):430-41.
    PMID: 21399583 MyJurnal
    The major outer membrane protein (OmpH) of 4 local Malaysian strains of Pasteurella multocida serotype B:2 were characterized in comparison to ATCC strains. Three major peptide bands of MW 26, 32 and 37 kDa were characterized using SDSPAGE. Two of these fragments, the 32 kDa and 37 kDa were observed to be more reactive with a mouse polyclonal antiserum in all of the local isolates as well as the ATCC strains in a Western blot. However, the 32 kDa fragment was found to cross react with other Gram negative bacteria. Therefore, the 37 kDa OmpH was selected as vaccine candidate. The 37 kDa ompH gene of the isolated strain 1710 was cloned into an Escherichia coli expression vector to produce large amounts of recombinant OmpH (rOmpH). The 37 kDa ompH gene of strain 1710 was sequenced. In comparison to a reference strain X-73 of the ompH of P. multocida, 39bp was found deleted in the 37 kDa ompH gene. However, the deletion did not shift the reading frame or change the amino acid sequence. The rOmpH was used in a mice protection study. Mice immunized and challenged intraperitoneally resulted 100% protection against P. multocida whilst mice immunized subcutaneously and challenged intraperitoneally only resulted 80% protection. The rOmpH is therefore a suitable candidate for vaccination field studies. The same rOmpH was also used to develop a potential diagnostic kit in an ELISA format.
    Matched MeSH terms: Molecular Weight
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