Without USP15, K63 ubiquitin piled onto Mtb-associated structures, LC3 recruitment increased, and bacterial replication was reduced. Blocking autophagy initiation removed this benefit, showing the effect depends on canonical autophagy.

Enjoyed celebrating John Neff’s PhD graduation ceremony yesterday.

PGL triggers rapid extracellular ATP release from host cells through pannexin channels. ATP then binds neuronal P2X3 purinergic receptors. Blocking P2X3 with a specific antagonist inhibits PGL-mediated neuronal activity. ⚠️. #anti-tussive 10/14

PGL is also made by other mycobacteria, like M. bovis, M. leprae, M. marinum and M. kansasii. By organically synthesizing PGL analogs, we found that neuroactivity is proportional to saccharide chain length & structure. Mtb PGL triggers the most robust activity with the lowest EC50, 30 pM! 9/14

We used whole body plethysmography in guinea pigs (the best model of human cough reflexes) to show that PGL is sufficient to induce cough 🚀. #cough #plethysmography #physiology 7/14

Together with @tedpricethepainguy.bsky.social’s lab at @utdpaincenter.bsky.social, we also found through single cell recording in human nociceptive neurons that PGL treatment raises the resting membrane potential, thereby increasing excitability. #electrophysiology 6/14

We used live cell calcium imaging in mouse and human nociceptive neurons to show that PGL alone → robust neuronal firing. Shown here a representative live cell calcium imaging experiment with a nociceptive neuronal cell line. Cells become green once activated → increased intracellular calcium. 5/14

Mtb HN878 is an isolate of the Mtb lineage 2, which some studies how shown is more transmissible than other Mtb lineages. Intriguingly, the commonly-studied lineage 4 strains have a frameshift mutation in a key polyketide synthase, rendering them unable to make PGL. #Aerobiology #Evolution 4/14

Celebrating the next PhD to leave the lab. Congratulations Dr. Neff. As per Shiloh lab tradition, John’s graduation cake represents Activity Based Proteomic Profiling with rock candy “proteins”.

First ever “Great Shiloh Lab Bake Off” was a big success. Loaf cake/bread was the assignment. Congrats to all 6 bakers and thanks to the 3 judges. Bakes were judged anonymously and comments were kind but fair.

Lab Secret Santa today was a big hit. Really enjoyed my gifts!

Celebrating the formal acceptance of Josephine’s paper in Journal of Clinical Investigation! In press proof to be released in two days!

Had a great time visiting Cornell University in Ithaca last Friday on a gorgeous and warm fall day. Thank you to Brian VanderVen for hosting my visit and for the campus tour.

Fantastic Levine Prize lecture yesterday by Noburo Mizushima from the University of Tokyo. He not only eulogized Dr. Beth Levine in the most elegant way possible, but also provided a great history of autophagy research & amazing cutting-edge & unpublished science.