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31.
Insert engineering and solubility screening improves recovery of virus‐like particle subunits displaying hydrophobic epitopes
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R. S. Abidin L. H. L. Lua A. P. J. Middelberg F. Sainsbury 《Protein science : a publication of the Protein Society》2015,24(11):1820-1828
The Polyomavirus coat protein, VP1 has been developed as an epitope presentation system able to provoke humoral immunity against a variety of pathogens, such as Influenza and Group A Streptococcus. The ability of the system to carry cytotoxic T cell epitopes on a surface‐exposed loop and the impact on protein solubility has not been examined. Four variations of three selected epitopes were cloned into surface‐exposed loops of VP1, and expressed in Escherichia coli. VP1 pentamers, also known as capsomeres, were purified via a glutathione‐S‐transferase tag. Size exclusion chromatography indicated severe aggregation of the recombinant VP1 during enzymatic tag removal resulting from the introduction the hydrophobic epitopes. Inserts were modified to possess double aspartic acid residues at each end of the hydrophobic epitopes and a high‐throughput buffer condition screen was implemented with protein aggregation monitored during tag removal by spectrophotometry and dynamic light scattering. These analyses showed that the insertion of charged residues at the extremities of epitopes could improve solubility of capsomeres and revealed multiple windows of opportunity for further condition optimization. A combination of epitope design, pH optimization, and the additive l ‐arginine permitted the recovery of soluble VP1 pentamers presenting hydrophobic epitopes and their subsequent assembly into virus‐like particles. 相似文献
32.
Novel electrode structures for large scale dielectrophoretic separations based on textile technology
The use of dielectrophoresis (DEP) to date has mainly been limited to processing small volumes due to difficulties in the fabrication of microelectrodes over large surface areas. To overcome this problem a novel approach to the construction of micro-electrode arrays has been developed based on weaving. A plain weave cloth was made from 100 microm diameter stainless steel wires and 75 decitex polyester yarns. The stainless steel wires formed the weft, and were kept parallel and apart by a warp of flexible polyester yarns, with a gap of around 150 microm between the metal wires. The metal wires were alternately connected to earth and signal of an AC power source, and it was shown that it was possible to collect yeast cells suspended in deionised water at the metal wire surfaces by dielectrophoresis. The polyester yarn was also found to distort the electric field, creating further areas of electric field non-uniformity around the polyester yarns, further enhancing the capability of the system to attract cells. A 14 ml separation chamber was built from the cloth by alternately sandwiching perspex slabs and cloth together. The DEP chamber was able to effectively collect life yeast from a flow of suspended cells through the cloth using an applied field of 1 MHz at flow rates up to 5 ml min-1. However, some loss occurred due to sedimentation. Also, the chamber was able to separate dead and live yeast cells at 30 Vpk-pk, 2 MHz, with some cell loss due to sedimentation. 相似文献
33.
Kahraman C. Rendi T. Ahmadi Salam N. Aka . nce Abidin . Alver A. 《Journal of Evolutionary Biochemistry and Physiology》2022,58(2):410-417
Journal of Evolutionary Biochemistry and Physiology - The sympathetic nervous system (SNS) plays an important role in regulating the metabolic and secretory functions of white adipose tissue. In... 相似文献
34.
Shahrul Hisham Zainal Ariffin Nur Akmal Mohamed Rozali Rohaya Megat Abdul Wahab Sahidan Senafi Intan Zarina Zainol Abidin Zaidah Zainal Ariffin 《Cytotechnology》2016,68(4):675-686
Transplantation of stem cells requires a huge amount of cells, deeming the expansion of the cells in vitro necessary. The aim of this study is to define the optimal combination of basal medium and serum for the expansion of suspension peripheral blood mononucleated stem cells (PBMNSCs) without resulting in loss in the differentiation potential. Mononucleated cells were isolated from both mice and human peripheral blood samples through gradient centrifugation and expanded in α-MEM, RPMI, MEM or DMEM supplemented with either NBCS or FBS. The suspension cells were then differentiated to osteoblast. Our data suggested that α-MEM supplemented with 10 % (v/v) NBCS gives the highest fold increase after 14 days of culture for both mice and human PBMNSCs, which were ~1.51 and ~2.01 times, respectively. The suspension PBMNSCs in the respective medium were also able to maintain osteoblast differentiation potential as supported by the significant increase in ALP specific activity. The cells are also viable during the differentiated states when using this media. All these data strongly suggested that α-MEM supplemented with 10 % NBCS is the best media for the expansion of both mouse and human suspension PBMNSCs. 相似文献
35.
Faizan Naeem Razali Amirah Ismail Nurhayati Zainal Abidin Adawiyah Suriza Shuib 《PloS one》2014,9(10)
The polysaccharide fraction from Solanum nigrum Linne has been shown to have antitumor activity by enhancing the CD4+/CD8+ ratio of the T-lymphocyte subpopulation. In this study, we analyzed a polysaccharide extract of S. nigrum to determine its modulating effects on RAW 264.7 murine macrophage cells since macrophages play a key role in inducing both innate and adaptive immune responses. Crude polysaccharide was extracted from the stem of S. nigrum and subjected to ion-exchange chromatography to partially purify the extract. Five polysaccharide fractions were then subjected to a cytotoxicity assay and a nitric oxide production assay. To further analyze the ability of the fractionated polysaccharide extract to activate macrophages, the phagocytosis activity and cytokine production were also measured. The polysaccharide fractions were not cytotoxic, but all of the fractions induced nitric oxide in RAW 264.7 cells. Of the five fractions tested, SN-ppF3 was the least toxic and also induced the greatest amount of nitric oxide, which was comparable to the inducible nitric oxide synthase expression detected in the cell lysate. This fraction also significantly induced phagocytosis activity and stimulated the production of tumor necrosis factor-α and interleukin-6. Our study showed that fraction SN-ppF3 could classically activate macrophages. Macrophage induction may be the manner in which polysaccharides from S. nigrum are able to prevent tumor growth. 相似文献
36.
This study synthesizes a tethered surface-grafted poly(acrylic acid) with quartz crystal microbalance (QCM) surfaces and provides detailed analysis of their properties and application. A tethered polyelectrolyte brush of poly(acrylic acid) is generated by first covering the substrate with a plasma-polymerized allyl alcohol (pp-AA) film, changing the polymerization initiators (bromination), and then grafting through atom transfer radical polymerization (ATRP) of tert-butyl acrylate (t-BA); these initiators are immobilized on a surface and exposed to a monomer. Finally, we convert the poly(t-BA) brush into poly(acrylic acid) through hydrolysis. We use the QCM technique to measure configuration change of the tethered poly(acrylic acid) grafted chains with two different degrees of polymerization (DP=50,200) in aqueous solutions at three different pH values (4.0, 4.8, and 5.4). The tethered poly(acrylic acid) grafted QCM shows that repeatable frequency responses are induced by pH change of solution. These frequency responses of large DP for pH are 20 times larger than responses of lower DP for pH. The frequency response of antibody immobilization on tethered poly(acrylic acid) grafted QCM (DP=200) and its frequency response of immunoreaction are 10 times larger than conventional immobilization methods by cysteamine with glutalaldehyde coupling of the antibody. The tethered poly(acrylic acid) grafted QCM can increase the frequency response for pH, the immobilization amount of antibody, and immunosensor response. 相似文献
37.
Exploring cultural differences in wildlife value orientations using student samples in seven nations
Jacobs Maarten H Dubois Sara Hosaka Tetsuro Ladanović Vukan Muslim Huda Farhana Mohamad Miller Kelly K. Numata Shinya Ranaweerage E. Straka Tanja M. Weston Michael A. Abidin Zulkhairi Azizi Zainal 《Biodiversity and Conservation》2022,31(3):757-777
Biodiversity and Conservation - Understanding differences in the way people think about wildlife across countries is important as many conservation challenges transcend jurisdictions. We explored... 相似文献
38.
Norhaslinda Zainal Abidin Mustafa Mamat Brian Dangerfield Jafri Haji Zulkepli Md. Azizul Baten Antoni Wibowo 《PloS one》2014,9(12)
Poor eating behavior has been identified as one of the core contributory factors of the childhood obesity epidemic. The consequences of obesity on numerous aspects of life are thoroughly explored in the existing literature. For instance, evidence shows that obesity is linked to incidences of diseases such as heart disease, type-2 diabetes, and some cancers, as well as psychosocial problems. To respond to the increasing trends in the UK, in 2008 the government set a target to reverse the prevalence of obesity (POB) back to 2000 levels by 2020. This paper will outline the application of system dynamics (SD) optimization to simulate the effect of changes in the eating behavior of British children (aged 2 to 15 years) on weight and obesity. This study also will identify how long it will take to achieve the government’s target. This paper proposed a simulation model called Intervention Childhood Obesity Dynamics (ICOD) by focusing the interrelations between various strands of knowledge in one complex human weight regulation system. The model offers distinct insights into the dynamics by capturing the complex interdependencies from the causal loop and feedback structure, with the intention to better understand how eating behaviors influence children’s weight, body mass index (BMI), and POB measurement. This study proposed a set of equations that are revised from the original (baseline) equations. The new functions are constructed using a RAMP function of linear decrement in portion size and number of meal variables from 2013 until 2020 in order to achieve the 2020 desired target. Findings from the optimization analysis revealed that the 2020 target won’t be achieved until 2026 at the earliest, six years late. Thus, the model suggested that a longer period may be needed to significantly reduce obesity in this population. 相似文献
39.
Thirty milled rice samples were collected from retailers in 4 provinces of Malaysia. These samples were evaluated for Aspergillus spp. infection by direct plating on malt extract salt agar (MESA). All Aspergillus holomorphs were isolated and identified using nucleotide sequences of ITS 1 and ITS 2 of rDNA. Five anamorphs (Aspergillus flavus, A. oryzae, A. tamarii, A. fumigatus and A. niger) and 5 teleomorphs (Eurotium rubrum, E. amstelodami, E. chevalieri, E. cristatum and E. tonophilum) were identified. The PCR-sequencing based technique for sequences of ITS 1 and ITS 2 is a fast technique for identification of Aspergillus and Eurotium species, although it doesn't work flawlessly for differentiation of Eurotium species. All Aspergillus and Eurotium isolates were screened for their ability to produce aflatoxin and ochratoxin A (OTA) by HPLC and TLC techniques. Only A. flavus isolate UPM 89 was able to produce aflatoxins B1 and B2. 相似文献
40.
Idris Abd Ghani Hamady Dieng Zainal Abidin Abu Hassan Norazsida Ramli Nadia Kermani Tomomitsu Satho Hamdan Ahmad Fatimah Bt Abang Yuki Fukumitsu Abu Hassan Ahmad 《PloS one》2013,8(12)