Science History – Page 2 – The Big Room (and the little things in it)

It’s comin’ right for us!

(Click image for Wunderground.com’s interactive “Wundermap”…)

Actually, looking more closely at the picture there I think we may be slightly further west than it indicates, but it’s still very likely we’ll end up losing power for a while sometime tonight, but I still am going to try to get a post up for the Giant’s Shoulders blog carnival – I’ve got the paper picked out and read, I just need to do a write-up before I go to bed tonight. Currently it looks like the winds shouldn’t get TOO bad until late tonight/early-tomorrow-morning so we ought to have power long enough…

Yes, there’s a disturbingly severe hurricane “Comin’ right for us!” and I’m worrying about posting about a classic scientific paper before the blog-carnival deadline while I still have power. How’s THAT for dedication?

First we need to move stuff inside, though. I don’t want The Ascospore getting blown away…

“Ueber die isolirte Faerbung der Schizomyceten in Schnitt- und Trockenpraeparaten”

the 1884, #6 edition of 'Medical Progress', containing Hans Christian Gram's invention of the 'Gram Stain' The Giant’s Shoulders blog carnival is coming up in two days, and I just realized I still haven’t gotten a post up for it yet. So, here it is.

I put up some quick reviews of several classic microbiology-methods papers for the previous edition of this blog carnival, but didn’t actually get around to putting up the one for what is almost certainly the most well-known microbiology technique: “The Gram Stain”. So, this post is about it:

Gram HC: “Ueber die isolirte Faerbung der Schizomyceten in Schnitt- und Trockenpraeparaten”; Fortschritte der Medicin; 1884; vol 2, pp 185-189

That’s “Regarding the Isolational(?) Coloring of Schizomycetes in Cut- [i.e. tissue sections] and Dried Preparations” in “Medical Progress”. The translation hosted by the American Society for Microbiology uses the word “Differential” where I’ve put “Isolational” – which is probably not quite right either but it’ll have to do for now – but I’ll get to that in a moment.

If you’ve ever been exposed to microbiology labwork before, you’ve almost certainly done or at least watched a procedure referred to as a “Gram stain”. In brief, you smear your sample with bacteria on a glass slide and bake it on, then you dump some purple stuff on it, them some brown stuff, then you rinse it briefly with alcohol, then you dump on some pink stuff, and then rinse it in water and look at it under a microscope. Bacteria that stay the original dark purple-blue color of the original purple/brown stuff are considered “Gram Positive”, and those that don’t instead appear the pink color of the last stain, and are considered “Gram Negative”. Many textbook authors and microbiology instructors will breathlessly proclaim that the Gram Stain reveals two “fundamental” categories of bacteria, but I’ll spare you my rant about that.

Properly speaking, this isn’t actually Gram’s stain, as described in his original paper. The modern variations that we’re all taught in microbiology class were developed later, and I believe they are nowadays based mainly on Victor Burke’s 1922 paper on the subject[1].

Regarding the title of the paper: “schizomycete” is what they used to call most kinds of bacteria. “Mycete” meaning “fungus”, as bacteria were assumed to be “plants without chlorophyll” just like molds and mushrooms, and “Schizo-” meaning “split in two”, since bacteria reproduce by splitting into two cells rather than by producing spores like “other” fungi. I say “most” because things like cyanobacteria (“blue-green algae”) or Green Sulfur Bacteria would have been referred to as “Schizophyta” (“fission-plants”). What Gram was originally trying to do wasn’t to differentiate one kind of bacteria from another, either, but to make it easy to tell bacteria from from the nuclei of cells in bacteria-infected tissue.

For that matter, Gram was really metaphorically standing on the shoulders of Koch and Erhlich, as he was building on their technique for staining “tubercle bacteria” – that is, tuberculosis-causing members of the genus Mycobacterium. Gram mentions that you need to stain this type of bacteria for the “usual” 12-24 hours to make this work, incidentally, as opposed to a few minutes for other “schizomycetes”. This suggests that you are expected to have some idea of what you’re going to find before you use the stain, as opposed to the modern implementation which is supposed to tell you something about what kind of bacteria you’re finding.

Still, Gram does report that some bacteria take the stain and some don’t, giving us a preview of the “differential” character of the modern version. He specifically notes typhoid and some causes of bronchial pneumonia fail to hold the stain. Given that Typhoid Fever is caused by a strain of the “Gram-negative” butt-bacter Salmonella enterica, and there are a number of “Gram negative” bacteria as well as “Gram positive” that can cause pneumonia, this makes sense. He also does mention the use of Bismarck Brown R a.k.a. Vesuvine as a counterstain in order to make the nuclei of the infected cells brown in contrast to the dark blue of the infectious bacteria in the tissue.

For much of the century-and-a-quarter since Gram’s publication, the question of why the Gram stain works was thoroughly investigated, and even today I occasionally hear or read assertions to the effect that the Gram Stain isn’t well understood. I disagree with this just as I think its importance to bacterial identification is grossly overblown, and if you want to know why, I have a previous post all about why the Gram stain works and how we know. You may or may not also be interested in an older post regarding whether or not “acid-fast” bacteria like the ones that cause tuberculosis (which don’t stain at all when using the modern version of the Gram stain) are “Gram Positive” or not. As always, if you spot any errors or have any questions, please let me know…

[1] Burke V: “Notes on the Gram Stain with Description of a New Method.” J Bacteriol. 1922 Mar;7(2):159-82.

(Tap Tap Tap) Is this thing on?…

Move is underway. Here’s a quick update / test.

Cornelia and Monk(eymutt), the Laser Dogs

On the way home (to Texas, that is, with a load of furniture from House v1.0 in Idaho) we adopted a co-dog. Now Cornelia isn’t the only official dog in the house.

Yeah, I know, pretty frivolous stuff. What do you want – this is mainly a test post. You guys ARE seeing this post, right? (Please comment and let me know). I’ve finally found and jumped through the hoops necessary to migrate the bigroom.org website over to the new host. It’s also running on the old server in House v1.0 on the DSL line as well, but this post isn’t getting put on it. If I’ve done everything correctly, hosting for bigroom.org should now be handled by Eskimo North. Just a few other minor tweaks and I’ll be set to return to House v1.0 and shut down the old server so I can move it down here to Texas.

I’ve gone from about 4400ft elevation down to about 250ft, but the plastic bottles of Mountain Dew� Wine seem to have positive pressure, so it would appear there is still some live yeast left in there and the priming sugar is slowly doing its job. I’m pretty sure the benzoic acid is what’s slowing down the activity so drastically, but it hasn’t killed it off yet.

I’ve also got a couple of entries in mind already for the next The Giants’ Shoulders blog carnival, and hopefully some more crazed brewing project news to offer sometime. And, of course, the promised post on why benzoic acid works. Stay tuned.

“Antibiotic Susceptibility Testing by a Standardized Single-Disk Method”

Okay, one last post in the Classic Science Papers challenge before my time’s up:

Bauer AW, Kirby WM, Sherris JC, Turck M :”Antibiotic susceptibility testing by a standardized single disk method.” Am J Clin Pathol. 1966 Apr;45(4):493-6.

The “Kirby-Bauer” antibiotic susceptibility test is another standard method that you should cover in microbiology class. The method involves getting a pure culture of the bacteria you want to treat, and then growing it in a petri dish. By putting paper disks soaked with various anti-bacterial substances, you can identify which ones are most effective at killing (or at least stopping) the bacteria in question – for example if you’re trying to figure out what kind of antibiotic to give to the guy coughing up some unknown plague in your doctor’s office… The anti-bacterial substance that the paper is soaked in slowly diffuses into the area around it on the petri dish, getting more dilute the further it gets from the paper. You can then estimate how powerfully anti-bacterial the stuff is by how far from the paper the bacteria stop growing.

The authors here didn’t invent this trick. Not all antibiotic-susceptibility tests are “Kirby-Bauer” tests (the blurry picture there is of an experiment I did involving the beer ingredient hops, and is not a Kirby-Bauer test. Click the picture to go to my “Beer Cures Anthrax” post from long ago…). What this paper describes is a method that finally standardized this test. Instead of having to use multiple paper disks with different amounts of the same substance, the “Kirby-Bauer” test prescribes specific concentrations of each antibiotic, and specific nutrient agar formulations, and so forth, so that determining which antibiotic your mystery bug is best treated with can be done in a way that gives consistent results regardless of who is performing the test.

The method is regularly updated to account for new antibiotics, but is still referred to as the “Kirby-Bauer” antibiotic susceptibility test to this day. Incidentally, the American Society for Microbiology kindly hosts a reprint of this paper as a .pdf file, so you can read it yourself if you’d like.

(UPDATE 20110328: new URL for the reprint of the paper. Thanks, Alex S!)

“A simplified method of staining endospores”

One more for the “classic papers” challenge:

Schaeffer AB, Fulton MD: “A Simplified Method of Staining Endospores”; 1933; Science; 77; pg 194

If you take a microbiology lab, this is the endospore staining technique (or “technic” as they used to spell it) that you’ll practice. This is a nice, simple, one-page paper. Alice B. Schaeffer and co-author Mac Donald Fulton describe a few of the other variations on endospore staining techniques, then describe how they’ve further simplified what they felt was previously the simplest one, described by a Mr. Wirtz in 1908.

“Endospores” are a sort of “escape pod” for certain specific kinds of bacteria. Unlike spores formed by yeasts and molds, these are not reproductive – each bacterium only produces one thick-coated spore, into which it shoves it’s genetic material and a few vital enzymes to get itself going again later when the spore finds itself in favorable conditions.

Since only a few kinds of bacteria produce these endospores, if you see endospores in your unknown bacterial culture it goes a long way towards helping to identify the bacterial species, so having a simple method for staining your bacteria so that endospores are obvious under a microscope is helpful. (Of course, these days most of us would rather just get a 16s rDNA sequence with PCR, but never mind that for now…)

Evidently, Wirtz’s original method involved using Osmium Tetraoxide (“osmic acid”) to stick the bacteria to the slide before staining. Not only is that stuff poisonous, it’s also expensive. I found a site selling sealed glass ampoules containing 1 gram each of this stuff for $35.00 each. Schaeffer and Fulton’s method does away with this in favor of much cheaper and easier heat-fixing (just as is done with the Gram stain and others). They use the dye “Malachite Green” for the initial stain, and steam-heat the dye-covered bacterial slide a few times to sort of “cook” the dye into the thick-walled endospores if they are there. Rinsing then washes the dye out of everything but the endospores, and a light red dye (safranin) is added as a counterstain. The end result is that under the microscope you’ll see light-red bacteria. If any of them form endospores, you’ll be able to see them as smaller green dots – sometimes still bulging inside of bacterial cells, sometimes floating around freely having escaped from the now-empty bacterial cell.

The “Schaeffer-Fulton Endospore Stain” is pretty easy to do, though the occasionally messy steambath part can be annoying. The method is pretty resistant to errors, so it’s not too hard to get good results even if you’ve never done it before.

Incidentally, you can buy Malachite Green at many pet stores – it’s still used as a treatment for “ick” (Ichthyophthirius infestation) in tropical fish.

Hmmm…still a couple of hours before it’s not longer May – Perhaps I can throw in one last post before time’s up…

I guess now I find out how good or bad Magellan, inc. is.

A couple of months ago I traded up from my Garmen etrex Legend to a Magellan eXplorist 600. I wanted to get away from Garmin due to a couple of irritating limitations that they seem to cling to (like losing your timestamps if you save an individual track, lack of SD card support, and dropping NMEA data support “to support [their] software” [see the review in Make #1]). So far, I’ve been pretty pleased with the shiny new Magellan. Though their software (as usual) only works on Microsoft Windows, I didn’t buy it for the software, and GPSBabel handles the data conversion of the files just fine.

So, naturally, now that I’ve just started blogging with the track data I’ve been recording with it, it has died on me. Yesterday – worked fine. Today – won’t turn on or respond in any way.

The good news is that I DID register the thing online at Magellan corporation’s website, and it’s supposed to have a 1-year warranty. Support’s only available Monday-Friday, though, so I’ll have to wait until Monday to see a response to my support query. Wish me luck. I’ll post updates in case anyone is interested in how Magellan responds.

In other news, “BigC” followed up on my previous queries about digital microscopes and Linux, saying that the manufacturer is now offering a Linux-compatible version of their AM311S model (the Linux-compatible one is the “AMU311S”) by special-order. Only goes up to 200X magnification, though, so it’s not real useful for my microbiological purposes. I can’t say the manufacturer of these digital scopes is impressing me much, but BigC certainly seems to be going out of their way to give good service.

There’ll be at least one other post later today – I wanted to get some more “classic” microbiology papers blogged before the day was up. Kirby-Bauer or Schaeffer-Fulton?…

“A small modification of Koch’s plating method.”

Only two more days for the Classic Papers Challenge, so if I’m going to get any more up, I’d better get my butt in gear.

Here’s a nice easy one:

Petri, R. J.:”Eine kleine Modification des Koch�schen Plattenverfahrens.” Centralblatt f�r Bacteriologie und Parasitenkunde; 1887; Vol. 1, pages 279-280.

The American Society for Microbiology has a translation available online. It’s only about a page-and-a-half of relatively large type – check it out.

There’s a trick we microbiologists use for growing bacteria. You make a solid (but wet) surface that contains whatever nutrients the microbe (bacteria, archaea, yeasts, mold spores…) you’re interested in need, and then you spread a diluted mixture of the microbe on it. The idea is that since the surface is solid the microbes can’t move around too much, and at any spot where a single cell starts initially, a whole pile of that cell and it’s genetically-identical (non-sexually-produced) clone-children will form until it gets big enough to see without a microscope. This cell-pile is called a “colony”, and you can poke or rub it with a sterile object, then stick the object into a sterile nutrient source. The end result is you have a “pure” culture of microbes that are effectively genetically identical. The solid material could be a lot of things – I’ve seen references to using slices of potato – though these days agar-agar gel mixed with nutrients is the preferred substance.

Koch (that is, Robert Koch of “Koch’s Postulates” fame, not Ed Koch the former mayor of New York City) used gelatin (so, hey, here’s another thing you can do with your expired Jell-O®). He apparently used to have a stack of shallow bowls, and had to use a special pouring device to carefully dump the gelatin into each stacked bowl in turn, then cover the works with a bell jar in order to keep stuff from falling into them from the air and contaminating them.

This was kind of a pain to work with, so some clever guy named Julius came up with a modification of this method in 1887, using pairs of shallow dishes, one slightly larger than the other so that it could be turned upside down to use as a lid. Then, you don’t necessarily need the bell jar, and you don’t need to stack them so they’re easier to pour.

Julius Robert Petri’s idea was so useful that we still use it today. Oh, yeah, and they named the dish-and-lid combination after him.

How’s that for a “classic” paper?

Meanwhile, my “Mountain Dew� Wine” project is turning out to be substantially more educational and fascinating than I’d hoped. There seems to be a decent amount of information available on how benzoic acid affects yeasts. I intend to turn that into a post later, but first I’ll try to find at least one more old paper to post before tomorrow is over…

“They laughed at me! But I’ll show them all! AH, HAHAHAHA!”

Another T-shirt to add to my list of T-shirts I want.

I’m spending more hours shoveling my way through the books and papers and crap we’ve got up here at House v1.0, since if all goes well I’ll be making a brief run back down to Southeast Texas so we can sign the papers for House v2.0 down there, at which point we’ll be able to start actually moving. I sure hope this one goes through. Not only is it our third attempt to buy a house down there, but I’ve already identified a convenient location to build my “Intentional Food Microbiology” brewlab in it.

Since there’s no way I can afford to buy a -80�F freezer, I have an obvious interest in alternate means of preserving the yeast, mold, and bacterial cultures that I want to keep. To me, drying seems like the most desirable method when it’s feasible, since dried cultures should require the least amount of maintenance. After a several-month delay, I’ve finally gotten around to getting back in touch with the archivist at Brewer’s Digest to see about getting an old article on the viability of dried yeast cultures[1].

Speaking of old but useful scientific papers, there’s an extremely nifty challenge going on through the month of May (deadline: May 31st) over at “Skulls in the Stars” blog: find a classic scientific paper, read it, and blog about it.

“My �challenge�, for those sciencebloggers who choose to accept it, is this: read and research an old, classic scientific paper and write a blog post about it. I recommend choosing something pre- World War II, as that was the era of hand-crafted, �in your basement�-style science. There�s a lot to learn not only about the ingenuity of researchers in an era when materials were not readily available, but also about the problems and concerns of scientists of that era, often things we take for granted now!”

I think this is a brilliant idea – the classic papers often seem to be forgotten and often explain things that people seem to take for granted these days. I already mentioned my post about the Gram Stain (original paper published in 1884), though that post really talks more about what has happened with the Gram Stain over the last 125 years rather than only being about the original paper. There are a couple of other classic microbiology papers that I’m going to try to get to if I have time before the May 31st deadline arrives.

I also need to get some yeast activated and get my must processed – I’m hoping a brief boil will reduce the amount of a yeast-inhibiting substance in it. I’ll post more detail after I get it going.

[1] Wickerham LJ, AND Flickinger MH:”Viability of yeast preserved two years by
the lyophile process.” 1946; Brewers Digest, 21, 55-59; 65.

All this week: A topic important to secular and religious people alike

It’s not midnight here yet, I’m still on time!

Hello, “Just Science 2008” subscribers and everyone else. My life is insane at the moment but dagnabbit I’m going to do my best to get at least one post up on a scientific topic every day from today (Monday, February 4th) until Friday…

Today’s post is in the form of a gedanken experiment.

First, imagine the following:

Some “entities” existing somewhere

It doesn’t matter what “entities” you are imagining, whether they are products in a market setting, or data structures in a computer program, or topics of discussion on a news broadcast. All that matters is that there can be more than one of them.

A mechanism by which these “entities” are copied (and, optionally, also sometimes removed)

Products are manufactured or recalled, data structures can be copied or deleted, additional news anchors can be added to comment on a topic or conversely may shut up about them…

At least one mechanism by which changes can occur between or during copies

Product designs can be changed, a computer program may consult a “random number” generator and use it to make small changes in the data structure, scriptwriters may alter the news anchor’s teleprompter messages…

Some aspect of the “entities” that affects the rate at which they are copied (and/or, optionally, removed).

Demand by buyers in the market results in ramping-up of production, a computer program may perform some test or comparison of a data structure and use the result to determine how many copies of it to make (or whether or not to delete it), news topics that result in more people watching are repeated more often while those that people tune out from are dropped from the schedule…

What happens to this group of “entities” over time should be obvious. Taking the example of products in a market, producers introduce a variety of products (the group of “entities” in this example) and buyers examine their characteristics and, based on which ones they like, buy some of them. The producers observe which kinds of products are selling more and make more of those, while reducing or outright eliminating the production of those that aren’t selling well. Over time, a few of the kinds of products in this group which best fit the preferences of the buyers and the ability of the producers to make them. These products will dominate the market until the preferences of the buyers or the ability of the producers to produce them change [example: a shortage in the price of a particular material needed for a popular product].

You have most likely observed this process in the “news topic” context yourself, where it tends to happen much faster as “cheap and easy” news stories are happily picked up by news agencies to broadcast until people get sick of them and tune out.

This can all, hopefully, be understood as a purely logical outcome – a conclusion that universally and necessarily follows from the premises given. There should be nothing supernatural or even surprising here, is there?

So, now that you understand why and how evolution works (if you didn’t before), I can move on. (Incidentally, the part of the example above that describes a computerized system is actually referred to as a “genetic algorithm”.)

My purpose in starting with this is because it really and truly is fundamental to the topic that I expect to spend most of this week posting about, and which has been of vital importance to human culture and intellectual development for thousands of years. This most important subject involves such notable figures as Charles Darwin,St. Thomas Aquinas, Noted American Science-guy Benjamin Franklin, New England Puritan Cotton Mather and Quaker William Penn ,Hardcore Catholics like Pope John Paul II, Hardcore Athiests like PZ Myers, even famous religious figures like Jesus.

I refer, of course, to wine (and beer and other examples of ethanol production).

Okay, here’s the background: I just graduated with my B.S. in Microbiology, and I’ve got this whole “Hillbilly Biotech”/”Do-it-yourself”/”Practical Science” kind of thing going on in my interests. That being the case, I wondered what it would take to isolate, culture, and maintain my own yeast (and bacteria – more on that later) stocks from the environment rather than buying “canned” cultures – or at least play with the “canned” yeasts to create my own stocks. As I was poking around, though, I kept running into the same attitudes – namely that it’s “too hard” to do this, and although there are a number of people who advocate re-culturing canned commercial yeasts for a short time to save money, none of them think it’s feasible to do this for more than a couple of generations, at which point we are assured that you have to go buy it again or else “mutations” will inevitably appear and scary and mysterious “off-flavors” will result and the brewing police will come and throw you in jail for deviating from the archetype of whatever pre-defined style of wine or beer you’re trying to make. Or something like that. In any case, it’s because of this fear of “mutations” that I am starting out with this “evolution”-related post: in biological evolution, various forms of alterations in the genetic material are the “changes before or during copying” in the gedanken experiment above.

I didn’t buy it when people were telling me that it was “too hard” to learn how my computer works so that I could run Linux and should instead leave deciding what my computer should do to the “professionals”, and I’m not buying the same argument about commercial yeasts, either. If I felt that way, I might as well leave the rest of the complex technology of brewing to the “professionals” too, and consign myself to “Lite Beer” and “Thunderbird” for the rest of my life.

I’ve been spending much of the last few weeks perusing books, online articles, and scientific papers on subjects related to brewing in general and brewing yeasts in particular, and this should form the bulk of this week’s post topics, of not well beyond this week. Tomorrow I intend to start in on the actual process of culturing yeasts. Meanwhile, feel free to correct my no doubt horribly over-simplified explanation of evolutionary processes in the comments.

This blog does not exist

I say that because in order to exist I must have used my computer to type it in, but George Berkeley “proved” that material things don’t exist. No pictures either this post, because after all my camera doesn’t exist, either.

Okay, the fact that I’ve got a whole cluster of time-sucking school stuff last week and this week to deal with is also a factor in keeping the posts here sparse at the moment. Berkeley just happens to be one of them.

Berkeley was what I would call a “philosophical” Empiricist (whereas I would describe myself as a “practical” empiricist – hence the “Applied” in this blog’s “Applied Empirical Naturalism” subtitle. Put simply, empiricism means that knowledge comes from observation via the senses. I’m a practical kind of guy, and I don’t think this in any way invalidates the use of the intellect to infer additional (testable) knowledge from one’s observations beyond what is directly observed. Berkeley, on the other hand, is a solipsist: he claims that nothing exists unless it is perceived – or is a perceiver.

His argument is a little hard to follow. As best I can tell, he’s starting with a Descartes-like observation that the only thing we ever actually experience are sensory perceptions. In other words, we can experience and know about the sensation of “heat”, but this sensation is just an idea in our minds. Even if there were something “behind” the sensation of heat that was causing it, we could not know anything about it directly, since we only ever experience the sensation.

In a way that is still not entirely clear to me, Berkeley then seems to take the leap from Descartes-style “the only thing I can be certain of existing from my direct observations are ideas, and my mind which contains them” to “since there is no direct empirical basis for claiming the existence of anything else, matter cannot be said to exist”.

Berkeley then goes on to claim that since only minds and ideas exist, and since there are some ideas that seem to be imposed on him (like if he sticks the idea of a red-hot-poker up the idea of his left nostril, he will have the idea of excruciating pain whether he wants to or not), that therefore there must be some other mind from which these ideas come. From this, he makes the leap to claiming that there must be an “infinite” mind which contains all these other ideas, by which he means God�.

This also gives him a convenient explanation for things existing when nobody’s looking at them. See, God is always looking at everything, so nothing that exists is ever not being perceived.

Personally, I’m finding myself wondering if his argument also leaves open the possibility of an animistic reality instead. He claims that everything we experience (including “sensible things”, i.e. things we see, feel, smell, etc.) is just an idea, and an idea existing without a mind is absurd. Instead of postulating the existence of an “infinite” mind, though, wouldn’t the notion that anything that exists actually does, itself, have a mind (or “spirit” if you prefer) also satisfactorily explain how things can continue to exist even when nobody is observing them? Berkeley makes the claim that inanimate objects don’t have minds…but he gives no justification for this claim. I mean, he admits that he can’t directly observe other people’s minds (or the “infinite” mind either) and therefore can’t prove that anyone but him exists, but he never claims that other people don’t exist. So why couldn’t the continued existence of the fork that I ate dinner with be due to the fork’s own mind?

That “thump” you may have imagined hearing was probably Berkeley turning over in his grave. Berkeley was, after all, a Bishop, going through this whole philosophical exercise out of hatred of “skeptics” and “atheists”, and it amuses me to imagine how appalled he’d be to have his arguments used to support something that he probably felt only “heathens” and “savages” would consider…

Yeah, I know, not much of a post, but I’m a bit overloaded at the moment. Nonetheless, more to follow this week over the next few days, at least.