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221.
222.
Guided selection is a method of producing a human version of a rodent or any other non-human antibody. The process is a serial transition from rodent to human via rodent-human chimaerics, through to a panel of human antibodies with similar characteristics to those of the starting rodent antibody. The guided selection process can be undertaken using either phage display or ribosome display, and chimaeric antibodies can be made either in series or parallel, with or without the retention of the original rodent CDR3s. Guided selection has successfully been used for the generation of a number of human versions of rodent antibodies, including HUMIRA, an inhibitor of tumour necrosis factor-alpha which is approved for the treatment of moderate to severe rheumatoid arthritis in over 40 countries. 相似文献
223.
In vitro growth and differentiation of mammalian sensory hair cell progenitors: a requirement for EGF and periotic mesenchyme 总被引:11,自引:0,他引:11
The sensory hair cells and supporting cells of the organ of Corti are generated by a precise program of coordinated cell division and differentiation. Since no regeneration occurs in the mature organ of Corti, loss of hair cells leads to deafness. To investigate the molecular basis of hair cell differentiation and their lack of regeneration, we have established a dissociated cell culture system in which sensory hair cells and supporting cells can be generated from mitotic precursors. By incorporating a Math1-GFP transgene expressed exclusively in hair cells, we have used this system to characterize the conditions required for the growth and differentiation of hair cells in culture. These conditions include a requirement for epidermal growth factor, as well as the presence of periotic mesenchymal cells. Lastly, we show that early postnatal cochlear tissue also contains cells that can divide and generate new sensory hair cells in vitro. 相似文献
224.
225.
Kienzle N Olver S Buttigieg K Groves P Janas ML Baz A Kelso A 《Journal of immunology (Baltimore, Md. : 1950)》2005,174(4):2021-2029
Exposure to IL-4 during activation of naive murine CD8+ T cells leads to generation of IL-4-producing effector cells with reduced surface CD8, low perforin, granzyme B and granzyme C mRNA, and poor cytolytic function. We show in this study that maximal development of these cells depended on exposure to IL-4 for the first 5 days of activation. Although IL-4 was not required at later times, CD8 T cell clones continued to lose surface CD8 expression with prolonged culture, suggesting commitment to the CD8low phenotype. This state was reversible in early differentiation. When single CD8low cells from 4-day cultures were cultured without IL-4, 65% gave rise to clones that partly or wholly comprised CD8high cells; the proportion of reverted clones was reduced or increased when the cells were cloned in the presence of IL-4 or anti-IL-4 Ab, respectively. CD8 expression positively correlated with perforin and granzyme A, B, and C mRNA, and negatively correlated with IL-4 mRNA levels among these clones. By contrast, most CD8low cells isolated at later time points maintained their phenotype, produced IL-4, and exhibited poor cytolytic function after many weeks in the absence of exogenous IL-4. We conclude that IL-4-dependent down-regulation of CD8 is associated with progressive differentiation and commitment to yield IL-4-producing cells with little cytolytic activity. These data suggest that the CD4-CD8- cells identified in some disease states may be the product of a previously unrecognized pathway of effector differentiation from conventional CD8+ T cells. 相似文献
226.
This paper reports the first year results of field experiments to determine the survival times of pathogens in livestock manures during storage and following land application, using viable count methods. E. coli O157, Salmonella and Campylobacter survived in stored slurries and dirty water for up to three months, with Listeria surviving for up to three months. In contrast, all these pathogens survived for less than one month in solid manure heaps where temperatures greater than 55 degrees C were obtained. Following manure spreading to land, E. coli O157, Salmonella and Campylobacter generally survived in the soil for up to one month after application to both the sandy arable and clay loam grassland soils, whereas Listeria commonly survived for more than one month. These data are being used to develop guidelines on the management of manures to minimize the risks of pathogen transfer from animal manures to the human food chain. 相似文献
227.
We describe two convenient syntheses of rhizobactin-1021 (Rz), a citrate-based siderophore amphiphile produced by the nitrogen-fixing root symbiont Rhizobium meliloti-1021, and several analogs. Our approach features a singly amidated, tert-butyl-protected citrate intermediate that easily affords a variety of Rz analogs in the late stages of the synthesis. Structural modeling and the monolayer behavior of Rz and its metal complexes are consistent with a structural reorganization upon Rz-mediated iron chelation. 相似文献
228.
Barley aleurone cells contain two types of vacuoles. Characterization Of lytic organelles by use of fluorescent probes 总被引:10,自引:1,他引:9 下载免费PDF全文
Light microscopy was used to study the structure and function of vacuoles in living protoplasts of barley (Hordeum vulgare cv Himalaya) aleurone. Light microscopy showed that aleurone protoplasts contain two distinct types of vacuole: the protein storage vacuole and a lysosome-like organelle, which we have called the secondary vacuole. Fluorescence microscopy using pH-sensitive fluorescent probes and a fluorogenic substrate for cysteine proteases showed that both protein storage vacuoles and secondary vacuoles are acidic, lytic organelles. Ratio imaging showed that the pH of secondary vacuoles was lower in aleurone protoplasts incubated in gibberellic acid than in those incubated in abscisic acid. Uptake of fluorescent probes into intact, isolated protein storage vacuoles and secondary vacuoles required ATP and occurred via at least two types of vanadate-sensitive, ATP-dependent tonoplast transporters. One transporter catalyzed the accumulation of glutathione-conjugated probes, and another transported probes not conjugated to glutathione. 相似文献
229.
I Kingma G H Groves E R Smith J V Tyberg 《Canadian journal of physiology and pharmacology》1986,64(7):892-896
In eight open chest dogs we assessed the creep of the pericardium by measuring the increase in surface area of the pericardium, occurring after pericardial surface pressure (Ppe) was rapidly increased by inflating an air-containing balloon positioned between the pericardium and the left ventricular (LV) epicardium. We observed an increase in LV end diastolic pressure (EDP) of 3.6 +/- 3.4 mmHg (1 mmHg = 133.3 Pa) (p less than 0.05) (mean +/- SD) and a reduction in LV anteroposterior (AP) diameter of 8.8 +/- 6.1 mm (p less than 0.01), both of which were stable after 10 s. Mean Ppe increased 11.6 +/- 3.3 mmHg (p less than 0.001). Pericardial surface lengths at 45 and 135 degrees to the long axis of the LV were measured with two pairs of ultrasonic crystals attached to the outer surface of the pericardium. The beam of ultrasound travelling between each pair was directed parallel to the pericardial surface through a film of conducting medium. Initial increase in surface area (calculated as the product of two pericardial lengths) occurring during the first 15 s after balloon inflation was 5.8 +/- 2.5% (p less than 0.001). During the next 30 min, while mean pericardial pressure did not change, pericardial surface area increased another 2.8% (p less than 0.005). This time-dependent 2.8% increase in pericardial surface area (equivalent to an increase in volume of approximately 5%) is due to creep.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
230.
We have compared clones of Syrian hamster cells selected for the first amplification of the CAD gene with clones selected for further amplification. The large domain amplified initially was not reamplified as an intact unit. Instead, subregions were reamplified preferentially, and parts of the initial array were often lost. These events reduced the average amount of coamplified DNA accompanying each copy of the selected gene. The degree of amplification was small in the first step (about three extra copies of CAD per cell), but second-step amplifications to a high copy number (up to 60 extra copies per cell) occurred frequently. After several separate steps of amplification, highly condensed arrays that brought many CAD genes close together were formed. In striking contrast to the stability of these highly amplified arrays, the low-copy chromosomal arrays formed early were quite unstable and were often lost completely within 1 or 2 months of growth without selection. The results suggest that different mechanisms may be involved in the first step of amplification and in the later evolution of an already amplified array. 相似文献