Contradictory evidence for NAC?

Hi all

http://www.ncbi.nlm.nih.gov/pubmed/20888409<

I'm not a scientist by any means so I don't know whether I'm understanding this correctly.  However, this paper seems to suggest that NACi increase the infectivity of Chlamydia infectionsi, while inhibition of glutathione synthesis and, by extension, lower NAC consumption, helps to reduce it.

My guess is that the ability of NAC to help break the disulfde bonds of the EBs is a positive thing outside the host cell and potentially counterproductive inside it because it allows the Cpni to access the nutrients/organelles etc on which it feeds.

The trial involved C Trachomatis, not CPn, but I'm assuming the same would apply.

Hopefully, this can be explained away by a more informed member but I really believe we ought to examine counter evidence where it exists.

If you're taking antibioticsi the "counterproductive" part is probably just fine, since the antibiotics will kill the Cpni after it's emerged from its shell.

What Norman said...

But consider this as well. Glutathione is neccessary for cellular health. In fact depletion of glutathione can cause severe illness and even death. Acetaminophen depletes gluatathione in the liver and when taken in large doses creates severe illness that requires hospitalization in which case the treatment is NACi.

What I think that paper was trying to prove was that in fact glutathione is what breaks down the EB'si disulphide bonds. But it does not present a novel treatment approach as what you can do to a cell culture in vitro is not in this case what you can do in vivo.

Also it is noteworthy that NAC is taken by quite a lot of people for general good health. It is likely that it is taken because it reduces the Cpni infection noticeably in even healthier people. Also whey is taken in (literally) bucket loads by serious athletes. Like NAC whey promotes the maintenance of glutathione levels.

As to the initial "NAC flu" that many people encounter when they start taking NAC, this is in my opinion just a neccessary process of depleting excess EB's that have accumulated in host cells. Say a cell has been infected for many weeks and glutathione levels become depleted. The cell would end up with a larger number of EBs and when it dies of necrosisi/apoptosisi, this larger number of EBs would infect a larger number of cells. When you initially take NAC more RBs are suddenly created in some host cells. Most likely this leads to necrosis in many of these but within a week or two a new homeostasis is reached with a lower average number of EBs per infected cell which is a step in the process of getting well.

Also it is worth mentioning that the photo on this site showing what appears to be a dramatic number of EBs in a blood sample is by far the exception. In most people, even some that presumably had significant systemic Cpn infectionsi, finding what looks like an EB was a challenge. The individual from that photo was someone who was into all sorts of alternative medicine and may have found some supplement that interfered with Cpn's ability to infect host cells, thus causing a build up in the blood. It would be great to know if this was the case but that line of inquiry likely passed with the individual.

- Paul

Also I should mention, I do not want to seem to keep arguing that it is not possible for NACi to be helpful outside the cell. It likely is. I just believe its primary benefit is inside of infected host cells. It takes several hours for glutathione (which is relatively concentrated in cells) to break the disulphide bonds of EBs. Outside of cells the concentration of NAC is likely to be lower and it is likely that Cpni typically infects a new host cell in minutes.

So any help is worthwhile but in my opinion the greater benfit of NAC is in restoring glutathione levels and thus limiting the number of EBs in infected host cells. What really should be done (maybe it has and I am not aware of it) is to take two samples of blood and treat one with NAC and see if they are equally infectious. For that matter I can't think of any experiment in which blood was used to infect a cell culture. But surely this has been done?

- Paul

"As to the initial "NACi flu" that many people encounter when they start taking NAC, this is in my opinion just a neccessary process of depleting excess EB'si that have accumulated in host cells."

Your opinion? What is that worth? Which laboratory do you hail from? Do you have a degree in microbiology? Have you ever seen and treated patients?

D W - [Myalgia and hypertension">i (typically 155/95.) Began (2003) taking doxycycline and macrolide and later adding metronidazolei. No medication now. Morning BP typically 110/75]

David,

Not sure what has you so worked up. Or when it became appropriate to make ad hominem attacks on this site over people's opinions. I did write "in my opinion" as to try and avoid controversy. But really I am pretty sure about this.

Consider the many papers you and I have both looked at with EMs of Cpni. In many of those images there were maybe 4-5 RBs and sometimes as many as 20-100 EBs. Since it only takes 2-3 hours to convert from EB to RB in a healthy cell and Cpn stays in a RB state for 2-3 days, these images should not be possible. The only explanation I have for this is depletion of cellular stores of glutathione. And as my above example of acetaminophen points out it is not too difficult to deplete cellular glutathione.

Both Stratton and Hammerschlag had difficulty in starting cultures with too many EB'si. They both found that if they added all of the EBs at one time the cell culture would die. So they would add a smaller number every day to create a culture with a persistent infection. So considering this, it would seem probable that suddenly restoring glutathione levels (which NACi does) would cause these host cells with many EBs to die of necrosisi as large numbers converted to RBs simultaneously.

I any event I think we can agree that reducing the number of EBs in host cells is a positive direction for treatment.

As to the extracellular EBs that appear to be attached to RBCs. This might in fact be an equally important factor. This is what I recall, bearing in mind this was something like 15 years ago. The blood samples were obtained from a finger prick and could only be observed for 5-10 minutes as the intense light from the scope would burn up the sample. I only recall looking at 5-6 people's samples as there were not too many people interested in getting pricked. Of these only one showed the huge number as is shown on this site. But this is too small a sample to draw any firm conclusions.

There is no way to come up with a quantitative number for the others but if I had to make my best guess, it was roughly as difficult to find a WBC as what we thought were EBs. So that would still be a significant number of what appeared to be EBs. And presumably the EBs on RBCs could be killed with NAC.

- Paul

Odd, Paul, that you think asking someone their academic and medical credentials constitutes an ad hominem attack. 

D W - [Myalgia and hypertension">i (typically 155/95.) Began (2003) taking doxycycline and macrolide and later adding metronidazolei. No medication now. Morning BP typically 110/75]

David,

Stating "Your opinion? What is that worth?" is an attack. If I made that statement to you, you would likely have a meltdown. In fact you seem to be practically having one simply because I pointed out some things that you don't seem to want to hear. Or do you disagree?

And as further argument, I have always thought that Cpni's primary means of proliferation through the body was through WBCs. I think WBCs sometimes phagocytize EBs and then carry those EBs to various sites that they are recruited to that have damage/infection/irritation and then sometimes undergo apoptosisi and leave EBs at the site. This seems to explain why we believe there is often more infection in areas that get regular damage/infection/irritation (i.e. feet, joints, sinus, lungs). If the blood stream were the significant means of distribution, I would think we would see a more even amount of infection across the body and a little of all symptoms in everyone.

Just my non MD opinion...

- Paul

Paul, that last line of yours will have answered David's question, so I guess it will have pleased him.  What he really dislikes is people coming on this site and talking as though they have vast medical knowledge and experience when they don't.  There are new people here all the time and not everyone knows the Paul of old!...............................Sarah

Completed Stratton/Wheldon regime for aggressive secondary progressive MSi in June 2007, after four years, three of which intermittent.   Still improving bit by bit and no relapses since finishing treatment.

Sarah,

I don't believe this is a requirement for the site. In any event David should perhaps consider the arguments I made as they are likely correct. Even if you don't like the message it is bad form to shoot the messenger ;-)

Anyway here is some background on the scope and that crazy blood sample...

The scope used in those photos was a Bradford Microscope. It is an amazing piece of engineering capable of 30,000x light imaging. It has a very large separate light box and piped a very intense light via fiber optics to the sample. Then it had custom optics and finally a camera used in spy satellites capable of gathering as much light as possible. This allowed for incredible magnification for a light microscope.

Unfortunately the inventor promoted it for some questionable medical applications. He thought he was investigating Lyme and promoted the use of some questionable substances that were given via IV. He may in fact have been on to something as that blood sample came from him. So the hypothesis that he aciidentally came up with some agent that interferred with Cpni's entry into cells is not totally implausible.

Anyway what he was doing was not approved by the FDA and the guy had no medical background so when a couple of people died he was ruined. I think he passed away not long after that.

- Paul

Paul

I apologize to you. Though usually quite mild-mannered, I was quite harsh. 

You seem to have erased your reference to the Chinese spy satellites, though. Probably for the best.

D W - [Myalgia and hypertension">i (typically 155/95.) Began (2003) taking doxycycline and macrolide and later adding metronidazolei. No medication now. Morning BP typically 110/75]

David,

Thanks.

I did and I am sure it was for the best ;-)

BTW in the microscope discussion I did not mean to suggest thaty anyone believed that scope could actually magnify 30,000x. That was the claim but something like 1,800x is the maximum resolution possible with light microscopy because of the wavelength of light. Still it was an impressive scope for its time and even if much of the magnification was "empty", it still made viewing samples much easier.

- Paul

When inside cells and preparing to leave, EBs are being constructed by Cpni.  This is a different environment from when they enter, where the EB shell starts out complete and something has to open it -- something which, per the cited article, is at least partly glutathione.  When EBs are forming, whatever chemistry induces the making of disulfide bonds is presumably provided by Cpn.  In particular, the cell's glutathione content doesn't have to equal the RB-becoming-EB'si glutathione content, since the RB is separated from the cell by the inclusion membrane and glutathione doesn't pass through membranes freely.  So while this article says something interesting about EBs entering into cells, it doesn't necessarily imply anything about forming EBs.

Nor, for that matter, about "excess EBs that have accumulated inside host cells", which is an idea I've only heard from you, Paul.  The usual model is that RBs turn into EBs and then disperse from the host cell -- though admittedly I've never heard whether this always involves the host cell's death, or just usually does, or what.  An image of a cell having a few RBs and a lot of EBs doesn't contradict this model; it can be explained by saying that the image was taken near the end of the Cpn life cycle.  Now if you had an image of a cell that had ruptured and released its EBs, but had managed to survive, with an EB or two that had happened to get stuck in it, that'd really be telling.  (I'm not sure how such an image could be distinguished from a newly-infected cell, but maybe there's a way.)

I'm also not sure what the difficulty in establishing a persistent infection is supposed to prove: when you start with too many EBs, it stands to reason that they'd kill off your cell culture, since the vast majority of cells would be infected at the first round, and then the remaining ones would be infected by the EBs released from those cells, or perhaps damaged by detritus from those cells.

Norman,

Cpni inclusions annoy me. They are pretty rare in vivo so I choose to ignore them. In vitro they may be more typical because of some of the agents that are used to promote Cpn growth (i.e. cycloheximide) or perhaps a less challenging environment or nutrient rich medium.

- Paul

I wouldn't have understood that comment, except that I'd heard it explained better by Stratton: in experimental studies, you commonly see Cpni RBs all clustered together, in an "inclusion".  This is often regarded as normal, but really is the effect of using cycloheximide, which inhibits protein synthesis in the host cell, largely shutting it down: without cycloheximide the RBs distribute themselves through the host cell, commonly putting themselves near mitochondria (which of course supply the ATP they use as fuel; and in a typical host cell there are many mitochondria).  But despite this difference in positioning, I'd expect that each RB (or bunch of RBs) is surrounded by a membrane.

Norman,

You can't have it both ways. There are either inclusions or not. You can't not have inclusions but have a "bunch of RBs surrounded by a membrane" which is by definition an inclusion. My point of them annoying me is that they are discussed so much, (theoretically) interfere with treatment approaches, and yet they barely exist in vivo.

My brief theory on inclusions is that they are a luxury that Cpni have when they first infect a host (or a new area in an existing host). At that time, NO production is lower (down regulated immunei response) and glutathione levels are higher. As they begin infecting nearby cells and glutathione becomes depleted (more likely precursors, just simplifying) and NO production becomes upregulated, this may be an approach that uses more resources than is worthwhile.

- Paul

There's nothing self-contradictory about the idea that a single RB can be surrounded by an inclusion membrane of the same sort that surrounds multiple RBs; indeed, it'd be surprising if it weren't.  All the biochemical machinery that Cpni has for interacting with the host cell (e.g. type 3 secretion system) either has to be set up to deal with such a membrane or to deal with its absence; it'd be quite surprising if it all worked whether or not there was a membrane there.

Norman,

When an EB first infects a host or new area of an existing host, the immunei system would likely be down regulated in that area. This means NO used to kill intracellulari pathogens would be down regulated. Also stores of glutathione or glutathione precursers would not be depleted. These both lead to Cpni following a course that produces inclusions. Additionally in the newly infected area tryptophan would not be sequestered, making for a better environment to proliferate. So in this circumstance it would serve Cpn's interest to expend additional resources hiding from the host cell. It would also serve its interest to produce as many EB'si as possible in the initial infection. Both of these goals may be met by creating an inclusion.

After this first cell dies from apoptosisi/necrosisi and hundreds of EBs are released to infect nearby cells, the need to expend energy camouflaging its presence from the immune system is not as important as the cell mediated immune system would now be upregulated anyway. So rather then expend resources on an inclusion it begins directly replicating.

Anyway if this idea is correct it would be quite surprising to see the "mini inclusions" you are suggesting and have not been seen by researchers. Although I will grant you that in some cases where the immune system downregulates even though there is an active infection, some EBs might differentiate down the inclusion path. But my gut feeling is that this is a non beneficial adaptation in the more hostile, immune upregulated environment as it uses more resources and presumably slows growth and I can't think of any advantages that could not already be incorporated into its outer membrane.

Anyway it is just a theory and not one I have thought long about, but it seems to explain what is known without adding complexity.

- Paul

Paul, you're the one who's proposing something novel here: that Cpni, which is generally agreed to surround itself with an inclusion membrane when living and growing in cells, somehow doesn't in real life, only in researchers' experiments.  I'm willing to grant you (because I heard it from Stratton) that Cpn RBs might normally distribute themselves through a cell, rather than clustering together in an inclusion.  But (a) this is not something I'm relying on, just something I'm conceding in response to your sneering at inclusions, and (b) I wouldn't expect it to affect whether or not there's a membrane around the Cpn, just whether or not they're all located in one spot in the host cell.

It's not like membranes take much energy to grow: they are typically just a couple of molecules thick.  So to the extent that they are useful for "hiding from the host cell", Cpn should be producing them all the time, as I believe it does.

By the way, Stratton gave a good reason for RBs to disperse themselves in the host cell: in a normal cell (that is, without cycloheximide) the RBs are competing with the host cell for ATP.  ATP concentrations are greatest near the mitochondria, so that's where they migrate to.  With cycloheximide, much of the cellular machinery of the host cell is shut down and not using ATP, so the RBs can all just stay in one place and let the ATP come to them; they don't have to go out and try to grab it as soon as it emerges from mitochondria before it's used up by the host cell.

Oh, and what Cpn does in response to immunei system pressure has been pretty well studied: basically it shuts down and goes into what most researchers call the "persistent" state, though on this site the term "cryptic state" is often used.  Antibioticsi produce this state, and so does interferon-gamma.  There are a lot of papers on this, and the ones I've seen have never described anything strange happening to the inclusion: it just stays there throughout, with the Cpn RBs inside it going into and out of the cryptic state.  (Yes, most or all of those papers used cycloheximide; but although that is regrettable it doesn't mean their research should just be ignored.)

Hi Norman,

That inclusions are not typically seen in vivo is not novel. Aside from a very rare example here or there they are not observed or detected. So you proposing they always or typically exist in vivo is novel.

At this scale effiiciency is everything. And it would be highly inefficient to create proteins that are then incorporated in the outer membrane to begin constructing a redundant membrane. And the energy and resources needed to build that new membrane are not inconsequential. As a very smart guy here said (may be paraphrasing) nature at this scale is ruthlessly efficient.

It would be more efficient and probable to have a mechanism that up or down regulates creating these proteins as needed. It appears researchers have identified the inhibitors/promotors of this with experiments involving NO/glutathione. My guess is they already have a similar hypothesis and are proving it one step at a time.

Not sure about the mitochondria/ATP thing. Cpni have no means of motive force so they cannot choose where they go. Not going to argue that Cpn might thrive closer to mitochondria and perhaps over time that gets you to the same observation.

As to the persistent state, that develops after the RB state, although I would guess this happens much faster under stress from ABXi or NO. And the stress from either of those may be in some cases what keeps them from following the path of converting to EBs. Anyway I have some ideas on this that I may post soon. We are already getting in the weeds a bit here.

- Paul

Edit: the exact quote was actually "Evolution at this scale is ruthlessly efficient." and was made by D.W.

Hi,

I am new here. I am just starting my CAPi treatment for chlamydia. I first did the NACi test and got the NAC flu (sinusitis, laryngitis, a little sore throat + worsening of brain fog). At first, I could tolerate 1200 mg NAC a day (for about a week), but now I have trouble tolerating even 300mg. I get brain fog, dizziness, tachycardia and palpitations and sometimes anxiety. Much worse than azithromycine.

How can I tell if it is some form of NAC intolerance or it is a beneficial die off reaction? Should I continue at low doses - 150mg, 300mg daily or should I drop the NAC?

Could the bursting of the EBs produce some kind of cytokinei storm that is affecting my brain? Do the EBs contain some kind of antigens or endotoxinsi? Or might it be that it is actually the gluthathion killing the intracellulari pathogens that is causing these reactions?

And what do you think about erdosteine? Can I use it as an alternative to NAC?

Best,

jack

concerning the original first post in this thread, i found the following paper

http://journals.plos.org/plospathogens/article?id=10.1371%2Fjournal.ppat...<

Francisella tularensis uses gluthathion as a source of cysteine. It might be similar with the chlamydia.

Do you have a link to a scientific paper showing that "inclusions are not typically seen in vivo"?  Or if not a scientific paper, to any other sort of documentation?  I mean, because I sure haven't seen any such thing; so it was novel to me, at least, when I heard it from Stratton.  I don't particularly doubt him, but it'd still be nice to have some other source besides his (and your) word.

Likewise for "It appears researchers have identified the inhibitors/promotors of this with experiments..."; which researchers?  Have they published anything?  If so, links please.

As for "no means of motive force", Cpni doesn't have anything like flagellae, so they can't move fast, but then they don't need to.  There are quite a lot of ways of moving slowly; it's not a stretch to suppose that they might have one.

(And I still think membranes are cheap compared to what goes inside them.)

As for being "in the weeds", these are the sort of details that matter.

Hi Jacck, Im new here too, I will just share my experience with NACi. After the runny nose and a somewhat different cοugh from the lugs I experienced some constant headaches in the front of my head and some more discomfort/pain in the heart area, also some tigling in throat too. I don't know if all those symptoms could be attributed to NAC but I initially reduced and then stopped NAC for some time because I worried about the heart area pain/discomford, I restarted later with smaller doses again and now I'm on my way to 2400.

Diplopia, nystagmus, right facial weaknes, feeling VERY sick and fainting, chronic cough for more than 10 years, heart palpitations, skiped beats, heart area discomfort and pain, weak hands grip, minor pains in body parts, nausea, insomnia and many

Norman,

People like Dr. Stratton, who have been researching Cpni long enough to have looked at the early EMs, recall that inclusions were not seen. It was only later when Cpn were commonly grown in vitro that they started to become common. Anyway proving a negative is difficult but I will do so as soon as you prove you have stopped beating your wife ;-)

Here is a paper on NO/glutathione and inclusions: http://www.ncbi.nlm.nih.gov/pubmed/12634074<

And yes that is a pretty big stretch. It would be the most interesting Cpn finding in years.

- Paul

Paul, are you trying to imply that none of these "early EMs" ever made it into the scientific literature, so that's why you can't point to one?  Or do you think I should be the one to look for such evidence, as being the person in this debate who actually cares about documented evidence?  (I'm not entirely averse to that, but at the moment I don't think I quite have enough to narrow down the search enough.  Like, what would "early" be?  Any authors' names?) 

In any case, note that I'm the one who is being asked to prove a negative: if I look through the early literature and find no such EMs, you could always say that I haven't searched hard enough, while you need only point to one paper that does have such an EM.  Or, for that matter, one paper that describes the typical Cpni infection of a cell and characterizes it as something very different from what is called an "inclusion" these days -- that for instance says that RBs disperse through the host cell and associate with mitochondria.  Extra credit, of course, if you find a paper that actually states the point under real contention here, namely that those dispersed RBs lack the outer membrane that multiple-RB inclusions have -- a membrane which renders it doubtful that glutathione can freely pass into the inclusion.

Cool down and read a few short stories. http://www.davidwheldon.co.uk/short_stories.html<  Apart from being a doctor I am an award-winning novelist. The stories may amuse you.

D W - [Myalgia and hypertension">i (typically 155/95.) Began (2003) taking doxycycline and macrolide and later adding metronidazolei. No medication now. Morning BP typically 110/75]

I have a hypothesis about this result

http://www.ncbi.nlm.nih.gov/pubmed/20888409<

specifically about this observation: "In the first two cases, we observed a four- to sixfold inhibition of C. trachomatis infection, whereas in NACi-treated cells we detected an increase in the size of chlamydial inclusions"

These "increased in size chlamydial inclusions" might actually be the stressed, cryptic, persistent form of cpni. Read some papers about the persistent form, for example this review, page 278, Chlamydial persistencei<

https://core.ac.uk/download/files/310/10902918.pdf<

Chlamydial inclusions in case of stress: "Instead, the normally round,  0.5–1.0  um  RBs,  become  significantly enlarged and aberrant in shape, when viewed by electron  microscopy"

---

the hypothesis is that the "increase in size" is caused by the stress from the gluthathion, i.e. the glutathion is actually fighting the infection.

The Line of Works is my favourite but Memoirs of a Delivery Boy is a hoot and mostly true from when David was one, as you can see here! http://www.davidwheldon.co.uk/memoirs-of-a-delivery-boy.pdf< ............Sarah 

Completed Stratton/Wheldon regime for aggressive secondary progressive MSi in June 2007, after four years, three of which intermittent.   Still improving bit by bit and no relapses since finishing treatment.

Chlamydia pneumoniae and chlamydia trachomatis have no more similarities than vibrio cholerae and other vibrio types of which there are many...................Sarah

Completed Stratton/Wheldon regime for aggressive secondary progressive MSi in June 2007, after four years, three of which intermittent.   Still improving bit by bit and no relapses since finishing treatment.

By the way, I probably should add that I have sympathy with the point of view "I've seen these damn things myself with my own eyes; why should I have to prove them to you?"  It's just that (a) this is a bit frustrating, and (b) I'm not entirely sure it's Paul's point of view here; he's sort of hinted at it, but if there's one thing I've learned the hard way, it's not to take those sorts of hints.  Oh, and (c) the way one "sees" such electron micrographs, these days, is commonly on a computer, in which case it's easy enough to pass them along.

In any case, the general answer to "...why should I have to prove them to you?" is "So that I in turn can prove them to others.  You want your ideas to spread, right, not to go no further than the sound of your own voice?"

Hi,

I'm new here as well. I do now the NACi test on myself nad can tell you this. I'm not on CAPi yet nad am taking just the NAC. I'm taking it for about 5 weeks and so far can not tolerate more than 400 mg/day split in 2 doses (in the morning upon waking up and then at least 4 hours before going to bed). When I first started on 2x 600 mg per day, I had terrible reactions and was very sick, more than without the NAC. I feel that the dose of 2x 200 mg/day is now ideal for myself and it really helps. My cough is not to dry any longer and I feel relief in all ways. After some time, when I consider it appropriate, I will start increasing doses, but just very slowly, step by step. But after having read your comments here, I'm rather very careful with that and stick with the dose which I can tolerate. Have a nice day.

Success is journey not destination

Comment viewing options

Select your preferred way to display the comments and click "Save settings" to activate your changes.