118 research outputs found
One-Pot Enol Silane Formation-Mukaiyama Aldol Reactions: Crossed Aldehyde-Aldehyde Coupling, Thioester Substrates, and Reactions in Ester Solvents
Trimethylsilyl trifluoromethanesulfonate (TMSOTf) and a trialkylamine base promote both in situ enol silane/silyl ketene acetal formation and Mukaiyama aldol addition reactions between a variety of reaction partners in a single reaction flask. Isolation of the required enol silane or silyl ketene acetal is not necessary. For example, crossed aldol reactions between α-disubstituted aldehydes and non-enolizable aldehydes yield b- hydroxy aldehydes in good yield. In a related reaction, the common laboratory solvent ethyl acetate functions as both an enolate precursor and a green reaction solvent. When thioesters are employed as enolate precursors, high yields for additions to non-enolizable aldehydes are routinely observed
Synthetic white balancing for intra-operative hyperspectral imaging
Hyperspectral imaging shows promise for surgical applications to
non-invasively provide spatially-resolved, spectral information. For
calibration purposes, a white reference image of a highly-reflective Lambertian
surface should be obtained under the same imaging conditions. Standard white
references are not sterilizable, and so are unsuitable for surgical
environments. We demonstrate the necessity for in situ white references and
address this by proposing a novel, sterile, synthetic reference construction
algorithm. The use of references obtained at different distances and lighting
conditions to the subject were examined. Spectral and color reconstructions
were compared with standard measurements qualitatively and quantitatively,
using and normalised RMSE respectively. The algorithm forms a
composite image from a video of a standard sterile ruler, whose imperfect
reflectivity is compensated for. The reference is modelled as the product of
independent spatial and spectral components, and a scalar factor accounting for
gain, exposure, and light intensity. Evaluation of synthetic references against
ideal but non-sterile references is performed using the same metrics alongside
pixel-by-pixel errors. Finally, intraoperative integration is assessed though
cadaveric experiments. Improper white balancing leads to increases in all
quantitative and qualitative errors. Synthetic references achieve median
pixel-by-pixel errors lower than 6.5% and produce similar reconstructions and
errors to an ideal reference. The algorithm integrated well into surgical
workflow, achieving median pixel-by-pixel errors of 4.77%, while maintaining
good spectral and color reconstruction.Comment: 22 pages, 10 figure
Sequential mutations associated with adaptation of human cytomegalovirus to growth in cell culture
Mutations that occurred during adaptation of human cytomegalovirus to cell culture were monitored by isolating four strains from clinical samples, passaging them in various cell types and sequencing ten complete virus genomes from the final passages. Mutational dynamics were assessed by targeted sequencing of intermediate passages and the original clinical samples. Gene RL13 and the UL128 locus (UL128L, consisting of genes UL128, UL130 and UL131A) mutated in all strains. Mutations in RL13 occurred in fibroblast, epithelial and endothelial cells, whereas those in UL128L were limited to fibroblasts and detected later than those in RL13. In addition, a region containing genes UL145, UL144, UL142, UL141 and UL140 mutated in three strains. All strains exhibited numerous mutations in other regions of the genome, with a preponderance in parts of the inverted repeats. An investigation was carried out on the kinetic growth yields of viruses derived from selected passages that were predominantly non-mutated in RL13 and UL128L (RL13+UL128L+), or that were largely mutated in RL13 (RL13−UL128L+) or both RL13 and UL128L (RL13−UL128L−). RL13−UL128L− viruses produced greater yields of infectious progeny than RL13−UL128L+ viruses, and RL13−UL128L+ viruses produced greater yields than RL13+UL128L+ viruses. These results suggest strongly that RL13 and UL128L exert at least partially independent suppressive effects on growth in fibroblasts. As all isolates proved genetically unstable in all cell types tested, caution is advised in choosing and monitoring strains for experimental studies of vulnerable functions, particularly those involved in cell tropism, immune evasion or growth temperance
The human cytomegalovirus ul11 protein interacts with the receptor tyrosine phosphatase cd45, resulting in functional paralysis of t cells
Human cytomegalovirus (CMV) exerts diverse and complex effects on the immune system, not all of which have been attributed to viral genes. Acute CMV infection results in transient restrictions in T cell proliferative ability, which can impair the control of the virus and increase the risk of secondary infections in patients with weakened or immature immune systems. In a search for new immunomodulatory proteins, we investigated the UL11 protein, a member of the CMV RL11 family. This protein family is defined by the RL11 domain, which has homology to immunoglobulin domains and adenoviral immunomodulatory proteins. We show that pUL11 is expressed on the cell surface and induces intercellular interactions with leukocytes. This was demonstrated to be due to the interaction of pUL11 with the receptor tyrosine phosphatase CD45, identified by mass spectrometry analysis of pUL11-associated proteins. CD45 expression is sufficient to mediate the interaction with pUL11 and is required for pUL11 binding to T cells, indicating that pUL11 is a specific CD45 ligand. CD45 has a pivotal function regulating T cell signaling thresholds; in its absence, the Src family kinase Lck is inactive and signaling through the T cell receptor (TCR) is therefore shut off. In the presence of pUL11, several CD45-mediated functions were inhibited. The induction of tyrosine phosphorylation of multiple signaling proteins upon TCR stimulation was reduced and T cell proliferation was impaired. We therefore conclude that pUL11 has immunosuppressive properties, and that disruption of T cell function via inhibition of CD45 is a previously unknown immunomodulatory strategy of CMV
Investgations on flight trajectory optimisation and adaptive control
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