The clinical manufacture of antigen-specific cytotoxic T lymphocytes (CTLs) for adoptive immunotherapy is limited by the complexity and time required to produce large numbers with the desired ...function and specificity. The culture conditions required are rigorous, and in some cases only achieved in 2-cm wells in which cell growth is limited by gas exchange, nutrients, and waste accumulation. Bioreactors developed to overcome these issues tend to be complex, expensive, and not always conducive to CTL growth. We observed that antigen-specific CTLs undergo 7 to 10 divisions poststimulation. However, the expected CTL numbers were achieved only in the first week of culture. By recreating the culture conditions present during this first week-low frequency of antigen-specific T cells and high frequency of feeder cells-we were able to increase CTL expansion to expected levels that could be sustained for several weeks without affecting phenotype or function. However, the number of 24-well plates needed was excessive and cultures required frequent media changes, increasing complexity and manufacturing costs. Therefore, we evaluated novel gas-permeable culture devices (G-Rex) with a silicone membrane at the base allowing gas exchange to occur uninhibited by the depth of the medium above. This system effectively supports the expansion of CTL and actually increases output by up to 20-fold while decreasing the required technician time. Importantly, this amplified cell expansion is not because of more cell divisions but because of reduced cell death. This bioprocess optimization increased T-cell output while decreasing the complexity and cost of CTL manufacture, making cell therapy more accessible.
A sanitation method that could continually clean and disinfect the air and surfaces in a hatchery could provide a second layer of microbial reduction on top of routine cleaning and disinfection. A ...gaseous dry hydrogen peroxide (DHP) system has been used in other facilities for this purpose and could have potential for use in chicken hatcheries. Because the DHP is a true gas and can permeate through the entire hatchery space, contact with eggs during storage and incubation could potentially interfere with normal hatching processes. Therefore, the aim of this study was to evaluate the effects of the DHP system on hatching parameters and chick quality. A total of 3,960 hatching eggs were collected from an ∼40-week-old Ross 308 broiler breeder flock and distributed in 2 treatments: treated and nontreated. For the treated group, the egg cooler was cleaned, and 1 DHP generator was placed inside. Two other DHP generators were placed in the common area outside as well. Both areas were treated for 7 D before placement of eggs, and then eggs were collected and placed inside the cooler over a 4-day period. Eggs were then stored for an additional 3 D after the last collection. Dry hydrogen peroxide levels were recorded each day during storage. For the nontreated group, all DHP machines were removed from the cooler and external room, and the egg cooler was cleaned. Eggs were collected in the same way for the control group as the treated group. After storage, eggs were placed into a single stage Natureform incubator. The eggs exposed to DHP showed higher (P < 0.05) hatchability of fertile eggs and lower (P < 0.05) early embryonic dead than eggs from the nontreated group. No other parameters evaluated were different between groups. Based on this work, the DHP treatment of fertile eggs had no detrimental effect on any performance parameter, with potential positive effects seen on hatch of fertile eggs and early embryonic dead embryos.
Repeated checking has been demonstrated to lead to reductions in memory confidence in several previous studies using student and clinical samples. This process of reduced confidence in memory and ...detail for memory, are thought to arise from the inhibition of perceptual processing that develops during repeated checking. Our research investigated whether reduced memory confidence from repeated checking could be attenuated through the use of novel stimuli during the repeated checking task. Three groups were generated through random assignment of 65 undergraduate students. As seen in previous research, individuals who repeatedly checked a stimulus (a virtual stovetop) showed reduced memory confidence, vividness, and detail, when compared with individuals who repeatedly checked a different stimulus. A third group in which the colour of the repeatedly-checked stovetop changed every five trials showed no significant decline in memory confidence between the pre-test and post-test. Results suggest that increased memory distrust can be ameliorated through the use of stimuli with characteristics that are novel and distinctive. Findings are discussed in the context of the existing model of repeated checking and memory confidence, and implications for treatment methods are presented.
► Repeated checking has been demonstrated to reduce memory confidence. ► Repeated checking leads to increased familiarity with the stimulus, leading to reduced memory vividness and detail. ► The use of distinctive stimuli can attenuate reductions in memory confidence that occur as a result of repeated checking.
Respiratory syncytial virus (RSV) is the cause of one-fifth of all lower respiratory tract infections worldwide and is increasingly being recognized as representing a serious threat to patient groups ...with poorly functioning or immature immune systems. Racemic 1,4-benzodiazepines show potent anti-RSV activity in vitro. Anti-RSV evaluation of 3-position R- and S-benzodiazepine enantiomers and subsequent optimization of this series resulted in selection of a clinical candidate. Antiviral activity was found to reside mainly in the S-enantiomer, and the R-enantiomers were consistently less active against RSV. Analogues of 1,4-(S)-benzodiazepine were synthesized as part of the lead optimization program at Arrow and tested in the XTT assay. From this exercise, (S)-1-(2-fluorophenyl)-3-(2-oxo-5-phenyl-2,3-dihydro-1H-benzoe1,4-diazepin-3-yl)-urea, 17b (RSV-604) was identified as a clinical candidate, exhibiting potent anti-RSV activity in the XTT assay, which was confirmed in secondary assays. Compound 17b also possessed a good pharmacokinetic profile and has now progressed into the clinic.
Respiratory syncytial virus (RSV) is the cause of one-fifth of all lower respiratory tract infections worldwide and is increasingly being recognized as a serious threat to patient groups with poorly ...functioning immune systems. Our approach to finding a novel inhibitor of this virus was to screen a 20 000-member diverse library in a whole cell XTT assay. Parallel assays were carried out in the absence of virus in order to quantify any associated cell toxicity. This identified 100 compounds with IC50's less than 50 μM. A-33903 (18), a 1,4-benzodiazepine analogue, was chosen as the starting point for lead optimization. This molecule was moderately active and demonstrated good pharmacokinetic properties. The most potent compounds identified from this work were A-58568 (47), A-58569 (44), and A-62066 (46), where modifications to the aromatic substitution enhanced potency, and A-58175 (42), where the amide linker was modified.
Here we present the Transcription Factor Encyclopedia (TFe), a new web-based compendium of mini review articles on transcription factors (TFs) that is founded on the principles of open access and ...collaboration. Our consortium of over 100 researchers has collectively contributed over 130 mini review articles on pertinent human, mouse and rat TFs. Notable features of the TFe website include a high-quality PDF generator and web API for programmatic data retrieval. TFe aims to rapidly educate scientists about the TFs they encounter through the delivery of succinct summaries written and vetted by experts in the field. TFe is available at http://www.cisreg.ca/tfe .
It is theorized that enhanced thermal heating may result from exposing single-walled carbon nanotubes （SWNTs） embedded in a conductive host to radiofrequency （RF） electric fields. We examine the ...RF-induced （13.56 MHz） heating behaviors of 95% metallic- and semiconducting-enriched SWNTs （m-/s-SWNTs） suspended in aqueous solutions with varying NaC1 molarity （0.001 mM-1 M）. The heating effects were only evident for host molarities below 1 mM （equivalent to 0.1 S/m） at which the s-SWNT heating rates dominated those of the m-SWNTs. The heating effects were localized to aligned and aggregated ＂SWNT ropes＂ -1 cm in length that formed in suspension, parallel to the electric-field vector, during the RF exposure. For molarities above 1 mM, no enhancements were evident, owing to the large heating effects of the bulk ionic NaC1 suspensions, which were observed in previous studies. Although larger enhancement effects proportional to the host conductivity have been theoretically predicted for m-/s-SWNT suspensions, this was not observed most likely because of the aggregation and screening effects, which diminished the scattered electric field near the m-/s-SWNTs. Our research may further the development of better nanoparticle heating agents for applications such as non-invasive RF-induced cancer hyperthermia.
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