A: - It's Indiana Jones in a petri dish.

[Hank & Don chuckle]

A: So it's, it's really-. But Don mentioned testing. So, here you have a thousand compounds in a flask - 

H: Mm-hmm?

A: You know, we don't want to isolate all of them because some of them might be -

H: Oh, yeah.

A: - something that's not very active or interesting.

H: Well, I mean, most of it is going to be just, like, "metabolism stuff" and -

A: Just-, yeah.

[all together]
A: Just like general-, exactly!
H: - "cell-membrane stuff" and  -
D: We're all made of-, yeah, things we're all made of.

A: A little DNA, a little sugar, -

H: Yeah.

A: So, when he mentioned "bioassay," the most important thing that drives the research is selecting the test that directs your discovery.

H: Mm-hmm.

A: So, when we started looking at the Berkeley Pit-. So this is not a very compatible ecosystem.

H: Mm-hmm.

A: So, you're looking at law pH - pH 2.5 -

H: Super acidic.

A: Think of, like, a Mountain Dew, so - nasty.

H: [taken aback] I - [shrugs in good-natured dismay]

[A&D laugh]

H: I mean, they might-they might, they might be a sponsor one day.

A: They could, oh, that's true -

H: [laughs]

A: - just like lemon juice.

H: Okay.

A: Think of lemon juice, orange juice.

H: Tight.

A: But they're are also very high in what are called "reactive metals" -

H: Mm-hmm.

A: - so, things like copper and iron. And so, these are generating -

H: Which is super-antibacterial, like, -

A: Abso-.

D: Yeah.

H: - bacteria hate metals like that.

A: If you were looking at copper sulfate, that's actually what they treat things with -

H: Yeah.

A: - to kill fungi.

H: Mm-hmm.

D: But they also kill us. [laughs]

H: Yeah.

A: And they-, yes.

H: Also bad for people.

A: Also bad for fish, so you'll never find fish in the Berkeley Pit because of the copper.

H: No, and the birds land in it and don't ever take off again.

D: Mm-hmm.

A: And they-, very few of them do successfully. But, what is it about those systems that are going to drive the chemistry these fungi elaborate? And so it turns out, some of these, sort of what we'd call "toxic" conditions in the pit mimic toxic conditions in mammalian systems!

H: Oh, like in the gut.

A: So if you think about inflammation - that's an acidic phenomenon.

H: Mm-hmm?

A: If you think about, "Reactive-oxygen species are produced a lot in metabolism, and they can lead to things like DNA mutation and damage."

A: - a lot of nuclear magnetic resonance and mass spectrometry, - 

H: Mm-hmm.

A:  - and Don looked at the NMR and it was completely different -

H: When there were two of them.

A: - than either fungus.

H: [quietly] Weird.

A: So then we started testing, and it turned out -

H: I'm gonna-, okay -

A: - active advanced bacteria.

H: I'm gonna go back to - ... sorry, I interrupted -

A: It's okay, no problem.

H: I'll let you-I'll let you get back to that, but first -

A: "Hold that thought."

H: - how are you doing NMR on... with so many compounds?

D: Well, like I said, the first thing you're gonna do is purify the compound -

H: Right.

D: - that is responsible for the activity that you're looking for.

H: Right, okay.

D: And once you purify, then you do NMR on those, so.

H: Because you need some pretty specific-, like you're not just gonna throw a soup into the NMR.

D: Well, we look at-, yeah, we look at crude extracts, too -

A: First.

D: - and they give us some information about what's the biggest components that's in that particular extract.

H: And so you can sort of tell sort of a difference between one organis-, like if I just blend one fungi and then another fungi, I can tell the difference in an NMR between those two things?

D: Yeah -

H: Wow.

D: - you could tell a difference in the the chemistry -

A: - in the chemistry.

H: Just that it's different. You couldn't necessarily tell, like, what compounds they are, but there's sort of a fingerprint there.

D: Yeah, there is a fingerprint.

A: And some of them you get to know, so you can actually look at a crude NMR and, "This looks like asperfuran, and this looks like-"

H: Interesting.

A: - penicillic acid." But that's because we look at these a LOT.

[H&D laugh]

A: These are our friends! We spend a lot of time with them. Youv'e got nerds on your couch here.

H: Yeah, no, that's what the couch is for.

D: Oh, good!

A: I felt the vibe, I felt very safe here.

H: So then you put these two fungi together, they were completely different, what did you find... Were they now good at something new?

A: Don came in with this beautiful new structure, and we looked at the structure and I said, "This reminds me of compounds like erythromycin," which is a "cyclic" - what's called - "macrolide." And these are very potent -

A: - and these are very potent antibiotics against gram-positive bacteria. Except things like erythromycin have a lot of sugars -

H: Mm-hmm?

A: - that are actually responsible for the activity. Well, our compound is sugar-free. 

H: Hm!

A: So, besides being -

H: No calories.

A: - less fueling, - 

[all laugh]

A: - it's not gonna harm your teeth! It didn't necessarily look like it could be antibiotic, so we have a wonderful colleague at University of Montana - Nigel Priestley, who's in chemistry - and he works at the discovery of antibiotics. And he, very graciously, had his technician, Jeremy, test our compounds and - it was incredible.

We were active only against gram-positives. Now, you know, you really want a gram-negative antibiotic, but we got a gram-positive one. 

H: Mm-hmm.

A: So different kinds of bacteria But what was really exciting we  were very active against Staphylococcus aureus. [slide] Now, Staph-aureus causes a lot of infections, soft-tissue damage, and other injuries. And they retested because they have a suite of what are called "Methecillin-Resistant Staphylococcus Aureus," what we call "MRSAs."

H: Yeah.

A: But we also use "MRSA" more generically to mean "a drug-resistant drug aureus."

H: Mm-hmm.

A: So they have a number of multi-drug resistance that they got from a research lab back east, and when they tested our compound against them, we were actually more active against drug-resistant staph than we are against a more generic variety.

H: Hm!

A: And when they tested again, we're actually AS active as vancomycin, ciprofloxacin, and 32-times more active than erythromycin - because a lot of these compounds have become resistant. 

H: Yeah, of course.

A: And the actual staph that are now resistant to erythromycin, we're still killing.

H: Mm-hmm.

A: And we've tested and retested, -

A: We made it onto a site; one of our students told us, "You're on Reddit." [Hank laughs] And I had NO idea what that meant.

H: Congrats on being on Reddit.

A: "You have 100-thousand points or something - [H wheezes] I did not even know what meant [Don chuckles], but he said, [A gives double thumbs-up sign] "It's cool."

H: It's good, it's a good thing.

A: I think so; we've done [The] Daily Show, Radiolab, and Reddit.

H: And you've taken a step down to SciShow Talk Show.

[D laughs]

A: No! This to be the pinnacle, this is exciting!

H: Well, we could talk forever about this, and you've found these microbes all over the place in weird spots, I mean, do you th-, are you particularly excited by the pit because there are weird things there? It's-it's like you end up with a, you know, a stranger group of organisms?

D: Ah, yes. And, well, we both started as the natural products chemists looking at marine organisms. The downside of that was you had to go to Bermuda, -

A: It was a tough duty.

H: Aw, I'm so, so sorry

D: - you had to go to the Bahamas to collect all of our samples. [all laugh]

H: In this one, you get to stay in (?~15:08)!

D: There you go, well, we ran out of money. [laughs] And you know, the pit was there, we found out that there were things growing in there, and we decided to look at 'em.

H: Nice.

A: We had a wonderful colleague - who was a chemist - who has since passed away, who actually in summer of 1995 brought us a stick coated in green slime that was in the pit. And that was the first -

H: It's like, "What is this thing that is alive."

A&D: Exactly!

D: Something was growing in the water.

A: And the [Montana] Bureau of Mines gave us a cup of water, and we started finding fungi. And what was really special about 1995 - that was the first November that snow geese landed in the pit during a storm and 342 snow geese died. And it was awful and -

H: Mm-hmm.

A: - some of the birds, quite a few of them, were not able to take off.