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351.
Elena Costariol Marco Rotondi Arman Amini Christopher J. Hewitt Alvin W. Nienow Thomas R.J. Heathman Martina Micheletti Qasim A. Rafiq 《Biotechnology and bioengineering》2019,116(10):2488-2502
Advanced cell and gene therapies such as chimeric antigen receptor T-cell immunotherapies (CAR-T), present a novel therapeutic modality for the treatment of acute and chronic conditions including acute lymphoblastic leukemia and non-Hodgkin lymphoma. However, the development of such immunotherapies requires the manufacture of large numbers of T-cells, which remains a major translational and commercial bottleneck due to the manual, small-scale, and often static culturing systems used for their production. Such systems are used because there is an unsubstantiated concern that primary T-cells are shear sensitive, or prefer static conditions, and therefore do not grow as effectively in more scalable, agitated systems, such as stirred-tank bioreactors, as compared with T-flasks and culture bags. In this study, we demonstrate that not only T-cells can be cultivated in an automated stirred-tank bioreactor system (ambr® 250), but that their growth is consistently and significantly better than that in T-flask static culture, with equivalent cell quality. Moreover, we demonstrate that at progressively higher agitation rates over the range studied here, and thereby, higher specific power inputs (P/M W kg−1), the higher the final viable T-cell density; that is, a cell density of 4.65 ± 0.24 × 106 viable cells ml−1 obtained at the highest P/M of 74 × 10−4 W kg−1 in comparison with 0.91 ± 0.07 × 106 viable cells ml−1 at the lowest P/M of 3.1 × 10−4 W kg−1. We posit that this improvement is due to the inability at the lower agitation rates to effectively suspend the Dynabeads®, which are required to activate the T-cells; and that contact between them is improved at the higher agitation rates. Importantly, from the data obtained, there is no indication that T-cells prefer being grown under static conditions or are sensitive to fluid dynamic stresses within a stirred-tank bioreactor system at the agitation speeds investigated. Indeed, the opposite has proven to be the case, whereby, the cells grow better under higher agitation speeds while maintaining their quality. This study is the first demonstration of primary T-cell ex vivo manufacture activated by Dynabeads® in an automated stirred-tank bioreactor system such as the ambr® 250 and the findings have the potential to be applied to multiple other cell candidates for advanced therapy applications. 相似文献
352.
Muhammad Shoaib B Sehgal Iqbal Gondal Laurence S Dooley Ross Coppel 《EURASIP Journal on Bioinformatics and Systems Biology》2009,2009(1):717136
While microarrays make it feasible to rapidly investigate many complex biological problems, their multistep fabrication has the proclivity for error at every stage. The standard tactic has been to either ignore or regard erroneous gene readings as missing values, though this assumption can exert a major influence upon postgenomic knowledge discovery methods like gene selection and gene regulatory network (GRN) reconstruction. This has been the catalyst for a raft of new flexible imputation algorithms including local least square impute and the recent heuristic collateral missing value imputation, which exploit the biological transactional behaviour of functionally correlated genes to afford accurate missing value estimation. This paper examines the influence of missing value imputation techniques upon postgenomic knowledge inference methods with results for various algorithms consistently corroborating that instead of ignoring missing values, recycling microarray data by flexible and robust imputation can provide substantial performance benefits for subsequent downstream procedures. 相似文献
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Amna Shoaib Mishaal Akhtar Arshad Javaid Haider Ali Zahra Nisar Shabnam Javed 《Physiology and Molecular Biology of Plants》2021,27(6):1361
The fungal pathogen, Alternaria alternata is responsible for causing leaf spot disease in many plants, including chili pepper. Zinc (Zn) an essential micronutrient for plant growth, also increases resistance in plants against diseases, and also acts as an antifungal agent. Here, in vitro effects of ZnSO4 on the propagation of A. alternata were investigated, and also in vivo, the effect of foliar application of ZnSO4 was investigated in chili pepper plants under disease stress. In vitro, ZnSO4 inhibited fungal growth in a dose-dependent manner, with complete inhibition being observed at the concentration of 8.50 mM. Hyphae and conidial damage were observed along with abnormal activity of antioxidant enzymes, Fourier-transform infrared spectroscopy confirmed the major changes in the protein structure of the fungal biomass after Zn accumulation. In vivo, pathogen infection caused the highest leaf spot disease incidence, and cumulative disease index, which resulted in a significant reduction in the plant’s growth (length and biomass), and physiochemical traits (photosynthetic pigment, activity of catalase, peroxidase, polyphenol oxidase, and phenylalanine ammonia lyase). The heat map and principal component analysis based on disease, growth and, physico-chemical variables generated useful information regarding the best treatment useful for disease management. Foliar Zn (0.036 mM) acted as a resistance inducer in chili pepper plants that improved activities of antioxidants (CAT and POX), and defense compounds (PPO and PAL), while managing 77% of disease. The study indicated foliar ZnSO4 as an effective and sustainable agriculture practice to manage Alternaria leaf spot disease in chili pepper plants. 相似文献
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Rouhollah Amini Atefeh Ebrahimi Adel Dabbagh Mohammadi Nasab 《Plant Species Biology》2021,36(2):338-347
Astrodaucus orientalis is a weed species in cropping systems and rangelands in Iran. The effects of temperature, light, NaCl concentration, water potential, seed burial depth and crop residue cover were assessed on seed germination and seedling emergence of two populations of A. orientalis from Ardabil (Meshginshahr population) and East Azarbayjan (Tabriz population) provinces of Iran. The A. orientalis populations indicated different responses to environmental factors and burial depth. In the Tabriz population the greatest germination (88.5%) was observed in 20/12°C day/night temperature but in the Meshginshahr population (83.2%) it was obtained in 24/16°C day/night temperature. Over a broad range of light period (10–24 hr light) germination was 74–83%, but it decreased (less than 37%) under 24 hr dark in both A. orientalis populations. With respect to water potential, the C50 parameters were −0.62 and − 0.49 MPa for Tabriz and Meshginshahr populations, respectively. The D50 parameters (the burial depth that caused 50% decrease in emergence) for Tabriz and Meshginshahr populations were 2.42 and 3.13 cm, respectively. Generally, the results showed that emergence of both populations of A. orientalis was delayed as depth of burial increased up to 4 cm and in cropping systems a shallow tillage that locates the seeds to >4 cm of depth in soil could be used in order to suppress seedling emergence. Our findings also could be useful in integrated management of A. orientalis in winter annual crops and rangelands. 相似文献
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