Type III secretory protein SINC of Chlamydia psittaci promotes host cell autophagy by activating the MAPK/ERK signaling pathway / 南方医科大学学报

OBJECTIVE@#To investigate the effects of SINC, a secreted protein of Chlamydia psittaci, on autophagy of host cells and the role of MAPK/ERK signaling pathway in mediating SINC-induced autophagy.@*METHODS@#RAW 264.7 cells treated with recombinant SINC were examined for changes in expression levels of LC3-II, Beclin-1, phosphorylated and total ERK1/2 using Western blotting. The expression level of LC3 in the treated cells was detected using immunofluorescence analysis, and the formation of autophagosomes and autolysosomes was observed with transmission electron microscopy (TEM). The effect of pretreatment with U0126 (a specific ERK inhibitor) on the expression levels of LC3-II and Beclin-1 in RAW 264.7 cells exposed to different concentrations of SINC was examined using Western blotting, and LC3 puncta in the cells was detected with immunofluorescence analysis.@*RESULTS@#The expression levels of LC3-II and Beclin-1 were the highest in RAW 264.7 cells treated with 2 μg/mL SINC for 12h. Immunofluorescence analysis showed exposure to SINC significantly increased the number of cells containing LC3 puncta, where the presence of autophagosomes and autolysosomes was detected. Exposure to 2 μg/mL SINC for 15 min resulted in the most significant increase of the ratios of p-ERK1/2/ERK1/2 in RAW 264.7 cells. Pretreatment of the cells with U0126 prior to SINC exposure significantly decreased the ratio of p-ERK1/2/ERK1/2, lowered the expression levels of LC3-II and Beclin-1, and decreased LC3 aggregation in the cells.@*CONCLUSIONS@#SINC exposure can induce autophagy in RAW 264.7 cells by activating the MAPK/ERK signaling pathway.

A New Tailored Sinc Pulse and Its Use for Multiband Pulse Design

PURPOSE: Among RF pulses, a sinc pulse is typically used for slice selection due to its frequency-selective feature. When a sinc pulse is implemented in practice, it needs to be apodized to avoid truncation artifacts at the expense of broadening the transition region of the excited-band profile. Here a sinc pulse tailored by a new apodization function is proposed that produces a sharper transition region with well suppression of truncation artifacts in comparison with conventional tailored sinc pulses. A multiband pulse designed using this newly apodized sinc pulse is also suggested inheriting the better performance of the newly apodized sinc pulse. MATERIALS AND METHODS: A new apodization function is introduced to taper a sinc pulse, playing a role to slightly shift the first zero-crossing of a tailored sinc pulse from the peak of the main lobe and thereby producing a narrower bandwidth as well as a sharper pass-band in the excitation profile. The newly apodized sinc pulse was also utilized to design a multiband pulse which inherits the performance of its constituent. Performances of the proposed sinc pulse and the multiband pulse generated with it were demonstrated by Bloch simulation and phantom imaging. RESULTS: In both simulations and experiments, the newly apodized sinc pulse yielded a narrower bandwidth and a sharper transition of the pass-band profile with a desirable degree of side-lobe suppression than the commonly used Hanning-windowed sinc pulse. The multiband pulse designed using the newly apodized sinc pulse also showed the better performance in multi-slice excitation than the one designed with the Hanning-windowed sinc pulse. CONCLUSION: The new tailored sinc pulse proposed here provides a better performance in slice (or slab) selection than conventional tailored sinc pulses. Thanks to the availability of analytical expression, it can also be utilized for multiband pulse design with great flexibility and readiness in implementation, transferring its better performance.