T3 intracellular calcium and caffeine

[Gingergrrl]

NDT is Armour?

Armour is NDT but my understanding (at least in the US) is that there are other supplement versions of NDT that you can buy (but I have never tried them and have only taken Armour since I was diagnosed with Hashimoto's in 2103).

If yes: Armour is NOT prescribed in Germany, not as a standard. You get it if it is prescribed (I think?), but nearly no doctor will prescribe it.

I wonder why it is not prescribed in Germany and it is very common here (and not expensive so insurance companies have no problems with approving it).

 

[Inara]

I wonder why it is not prescribed in Germany and it is very common here

Maybe because most German doctors are blind and closed minded? Just my personal experience - you can find the 5-10% good ones, but it's difficult.

 

[Inara]

The results from the calcium flux test should come this week. But why I write is because the test they performed is quite interesting. It goes back to a paper that was published in 2011:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3044412/
ORAI1-mediated calcium influx is required for human cytotoxic lymphocyte degranulation and target cell lysis

The test looks like this:

To analyze the consequences of this mutation for SOCE in NK cells, changes in intracellular Ca2+ levels were measured in purified NK cells following the passive depletion of intracellular Ca2+ stores. For this purpose, we either treated NK cells with thapsigargin (Fig. 1C, Lower), which depletes ER Ca2+ stores through inhibition of the sarco/endoplasmatic Ca2+ ATPase, or activated NK cells through cross-linking of CD16 (Fig. 1C, Upper), which causes emptying of ER Ca2+ stores via phospholipase C-γ activation and phosphoinositide generation. Freshly isolated NK cells were treated with thapsigargin or anti-CD16 mAb in the absence of extracellular Ca2+ (Fig. 1C). Readdition of Ca2+ to the medium led to robust Ca2+ influx in NK cells from healthy donors but not in NK cells from the patient, showing that ORAI1 is essential for SOCE in NK cells.

Here's a sketch about ORAI1, SERCA and IP3R from another paper, namely
"Regulation of epithelial ion transport in exocrine glands by store-operated Ca2+entry"
https://www.sciencedirect.com/science/article/pii/S0143416016302147

It's Fig. B.
SOCE seems to be the process of Ca transport from extracellular to intracellular?
Via ORAI1 Ca enters the cell (purple). SERCA is something like an "ATP driven pump" that pumps intracellular calcium into the ER (white). Via IP3Rs (and Ryanodine receptors, but they don't seem to play a role here) Ca leaves the ER into the cell. STIM1 is something like a sensor that recognizes if Ca in the ER (and intracellular?) is too low. If so, channels are opened to transport Ca from the outside into the cell and ER (here: OPAI1 and SERCA).

What they did:
NK cells were isolated and put into a medium with a flourescend substance that glows brighter the more calcium is there. Extracellular Ca was removed (I didn't find that intracellular calcium was removed, too, but it would make sense). Ca stores from ER were emptied via thapsigargin (which inhibits SERCA) and an antibody that activates PCL-gamma (phospholipase-C-gamma; PCL-gamma stimulates the production of IP3 - and DAG, if I remember correctly - which, together with Ca, binds to the IP3Rs and opens them). Then Ca was added to the external medium and the flux from extracellular to intracellular through OPAI1 was measured. At this time, SERCA was still blocked, and no Ca could enter or leave the ER. So the measured Ca flux seems to give reliable information about the function of OPAI1.

Their method seems to be suitable to judge the function of OPAI1, but not of the IP3Rs - only if all IP3Rs can't open so that Ca from ER can't flow, which would maybe happen if all IP3Rs are dysfunctional. In that case STIM1 wouldn't signal the need for Ca and OPAI1 wouldn't be opened. Why the ryanodine receptors are not mentioned I don't know - maybe they're not on the ER membrane in T cells and NK cells?

So, I would say, with their test no qualitative (or maybe even quantitative) statement can be made about IP3Rs. All one can say is, maybe, that Ca flux from ER into the cell seems to work - somehow.
What also must be considered is the expression of the different IP3Rs in different cells. If IP3R3 is minor in T cells/NK cells (I don't know), the other two types do all the work, and a dysfunction of IP3R3 won't have a huge effect.
Just thoughts from my side...

What's also interesting to me: Somehow plasma membrane channels, like voltage gated channels, didn't play a role or weren't addressed? Still more to understand...

But I need to see if it's exactly that test they applied to my blood.

So, at this point, I still can't say if the mutation in ITPR3 is significant or not. I need to look further.

Edit: I am not a biochemist, and the above mirrors my understanding from what I have read. It's possible some things are incorrect, and I welcome corrections and deeper insights.

 

[Inara]

Here's another picture:

https://www.researchgate.net/profil...-mechanism-of-CRAC-channel-activation-The.png

 

[Gingergrrl]

Maybe because most German doctors are blind and closed minded? Just my personal experience - you can find the 5-10% good ones, but it's difficult.

I've always thought the opposite (from things I read on the board) and some of the research and tests that were most helpful to my personal case were from Germany (Cell Trend Labs, Dr. Scheibenbogen's work, etc) but I know this is more on the research side vs. clinical care. Although my understanding is that Germany is one of the only countries that does heart biopsies to test for viruses (and other stuff). Armour is so benign and common here, it is fascinating that Germany would do heart biopsies but not prescribe Armour

The results from the calcium flux test should come this week.

I am excited for you that you will get your results soon!

What they did:

I understood exactly 0% of what you explained and yet I still found it interesting .

What's also interesting to me: Somehow plasma membrane channels, like voltage gated channels, didn't play a role or weren't addressed? Still more to understand...

So the test shows that the voltage gated calcium channels don't play any role in connection to your genetic mutation?

 

[Inara]

but I know this is more on the research side vs. clinical care.

Also, there is the difference private vs. non-private doctors. So if one only talks with people who have a private health insurance, you might think Germany has the best health system on earth. But only ca. 10% can afford a private insurance, the rest doesn't earn enough. And for 90% the health system is a disaster - with exceptions (like heart diseases).
If you pay privately, you can get nearly any tests you like, including the "good" ones, that look "deeper".

I don't think German research is excellent/good; there's not enough money going into research (in my opinion). But I think Scheibenbogen et al. do good research.

So the test shows that the voltage gated calcium channels don't play any role in connection to your genetic mutation?

If I remember correctly, voltage gated calcium channels are also positioned on the plasma membrane - does someone know better? In the OPAI1 paper, voltage gated calcium channels weren't mentioned - either because they're not part of T cells/NK cells, or because they don't play a role (or they forgot it, which I don't think).

I was just wondering if the test really looks at OPAI1 functionality alone, or that by neglecting other calcium channels (like voltage gated ones, or ryanodine receptors) the results could be skewed.
I also wondered if by looking on OPAI1 one can deduce something about IP3Rs, or the like, but I don't see how.
But most probably I lack the knowledge. Maybe in a cleansed medium with isolated cells voltages play no role because, e.g., there aren't currents.

 

[Gingergrrl]

Also, there is the difference private vs. non-private doctors. So if one only talks with people who have a private health insurance, you might think Germany has the best health system on earth. But only ca. 10% can afford a private insurance, the rest doesn't earn enough. And for 90% the health system is a disaster - with exceptions (like heart diseases).

This makes perfect sense and I suspect that the people I have talked to (or read their stories) from Germany had private insurance.

But I think Scheibenbogen et al. do good research.

Agreed.

If I remember correctly, voltage gated calcium channels are also positioned on the plasma membrane - does someone know better?

I assume this is correct but you know much more about all this than I do! I'm not replying to anything beyond that b/c I don't know the answers

 

[jjxx]

I think the initial high calcium also had something to do with the Vyvanse I was taking and Armour thyroid.

The current ratios are
Ca/Mg 13.6 (4-30)
Ca/P 5.33 (1-12)
Na/K 3.57 (0.5-10)
Zn/Cu 15 (4-20)

One other thing that has improved on this last test is phosphorus which was always on the low side but their interpretation guide says that it's not a marker of phosphorus but of cellular metabolism.

Could you let us know what test this is?

 

[Iritu1021]

The test is Hair Mineral Analysis by Doctor's Data. I have also written a blog post a few months ago about this test a while back.

Note: It won't be accurate it if you have dyed your hair.

 

[jjxx]

The test is Hair Mineral Analysis by Doctor's Data. I have also written a blog post a few months ago about this test a while back.

Note: It won't be accurate it if you have dyed your hair.

Thank you for your prompt response and I will certainly visit your blog.

 

[Inara]

@Gingergrrl, which calcium channels are affected in you? Was it the L-type?
I found this:
https://link.springer.com/chapter/10.1007/978-94-007-2888-2_44

In this chapter, we provide an overview of recent advances in our understanding of Ca2+ signaling in mast cells with a special attention to the emerging role for the L-type Ca2+ channels as a regulator of mast cell survival.

 

[BadBadBear]

Dandelion Root Extract Induces Intracellular Ca2+Increases in HEK293 Cells
http://www.mdpi.com/1422-0067/19/4/1112

Perhaps dandelion tea with milk as an alternative to coffee?

That is super interesting! When I was on T3 only I craved dandelion root, had it every day added to every cup of coffee I drank for about 3 years running. Now on mostly T4 with a small amount of Armour, I don't like it anymore. Wonder if this is why.

 

[Gingergrrl]

@Gingergrrl, which calcium channels are affected in you? Was it the L-type? I found this:https://link.springer.com/chapter/10.1007/978-94-007-2888-2_44

Hi Inara, the calcium channel that I have autoantibodies to is the "N-type". However, I don't believe that there is a test for autoantibodies to the L-type? The panels that I did from Mayo (in 2016 & 2018) test for the N-type and P/Q type but do not test for the L-type. I am positive on the N-type (both in 2016 and now) and negative on the P/Q type (both times).

I have never been tested for the L-type. Do you know if there is a test for it? I am curious b/c Propofol (anesthesia) blocks the L-type CA+ Channel and I have no idea if it would ever be safe for me to have this anesthesia b/c of that.

I read your link re: mast cells but didn't quite understand it . What is the connection between the L-type channel and mast cell activation (in the most basic terms)?

 

[Inara]

Hi Gingergrrl,

I didn't read the paper (it's behind a paywall), just the abstract. I just found interesting the connection between L-type calcium channels and mast cells. I already know that calcium signaling plays a role in the regulation of mast cells, and I hope to find details. I hypothesized (already months ago) that a disruption in calcium signaling may lead to mast cell issues, too, including mast cell activation.

I imagine calcium signaling a bit like digital signaling with "0" and "1" (that's oversimplified). Depending on the arrangement of 0 and 1, you have a different content. Eg. 1001 would say "get me protein 1", 1100 would say "activate receptor 2" etc. In calcium signaling, that would be "sinusoid signal", "monotonic increasing signal", "puffs" etc. In digital signaling, it is not uncommon to have errors. There are checking and correction processes - as often in calcium signaling. But if an error slips through, you have a different signal. And this could lead to problems. E.g. if your Calcium signal is falsely too strong (I'm just inventing), mast cells think they have to "activate", although they shouldn't. This in turn may lead to "feedback loops", and so mast cells are activated and activated, substances are set free which may lead to inflammation or whatever.

I don't know if one can be tested for L-type channels.

 

[Gingergrrl]

I just found interesting the connection between L-type calcium channels and mast cells. I already know that calcium signaling plays a role in the regulation of mast cells, and I hope to find details.

I find it very interesting, too, and have read many prior posts re: a connection between calcium channels and MCAS but never quite understood them. It seemed that some people with MCAS were helped by calcium channel blockers (if I am remembering correctly) but in my case, I am not supposed to take CCB's b/c of my LEMS/CA+ Channel blocking autoantibody which is a totally separate issue from MCAS as far as I know... but I don't know much

I imagine calcium signaling a bit like digital signaling with "0" and "1" (that's oversimplified). Depending on the arrangement of 0 and 1, you have a different content. Eg. 1001 would say "get me protein 1", 1100 would say "activate receptor 2" etc... But if an error slips through, you have a different signal. And this could lead to problems. E.g. if your Calcium signal is falsely too strong (I'm just inventing), mast cells think they have to "activate", although they shouldn't. This in turn may lead to "feedback loops", and so mast cells are activated and activated, substances are set free which may lead to inflammation or whatever.

That is really interesting (all the ways that calcium signaling could potentially go wrong).

I don't know if one can be tested for L-type channels.

I don't believe that there is (at least not a test that is commercially available to patients) but there must be one within research studies (?)

 

[Inara]

Dear @Gingergrrl, can you tell me (again) - are your calcium channels blocked due to the autoimmunity, or are they more "open"? Were you told not to take calcium channel blockers?

I ask because mast cell activation seems to need increased calcium levels in the cell. So, I wonder, maybe "leaking" calcium channels contribute to MCAS? This is just a guess!

 

[Gingergrrl]

Dear @Gingergrrl, can you tell me (again) - are your calcium channels blocked due to the autoimmunity, or are they more "open"? Were you told not to take calcium channel blockers?

Hi Inara! I don't mind you asking me again and I can explain it (to the best of my knowledge) but it would be so much better if I had a science background!

I can only assume that my (N-type) Calcium Channels are blocked b/c of the autoimmunity/ autoantibodies and don't think it is for any other reason. I don't know if they are any more "open" now with my remission of symptoms b/c the actual autoantibody level has remained (virtually) the same. It was 0.05 prior to treatment and it is now 0.06 (virtually the same) and anything over 0.02 is considered positive by Mayo Clinic's lab.

I was explicitly told not to take calcium channel blockers by three Neuros, including the original Neuro at Stanford who ran the tests that were sent to Mayo. I later saw two Neuros in my own city who said the same thing (to avoid all meds that were CCB's). One went so far as to say to avoid Magnesium b/c it is technically a CCB but the others felt that Mg was okay.

I hope this answered your question?!

I ask because mast cell activation seems to need increased calcium levels in the cell. So, I wonder, maybe "leaking" calcium channels contribute to MCAS? This is just a guess!

I have definitely heard that there is a connection between calcium and MCAS but I am not sure what it is. I am assuming it pertains to regular calcium vs. the ionized or voltage gated kind of calcium? I have seen websites in which people with MCAS were told that CCB's could help them (and I have no doubt that this is true in some cases) but can only assume that those people do not also have the LEMS/CA+ Channel autoantibodies like I do.

 

[frozenborderline]

Intravenous lidocaine is very effective for chronic and acute pain, so I have considered it, but oral ambroxol seems so much easier.

What amazes me is that these sodium channels blockers have both an immediate action on pain, and a delayed action on inflammation, and I need both....

I am also interested on Ambroxol because it was suggested it may have an effect on tinnitus.

The thing is it may not be a good association with T3, so I will see if I can stop it while on Ambroxol.

I want to make a whole separate post on this, but lidocaine and procaine have been found to demethylate dna and possibly have lasting (often beneficial) epigenetic changes. Very very interesting drug. The intravenous safety is in a very narrow range though so I wouldn’t do that on my own but would experiment w local injection for pain, use on muscles, even swallowing it. WIll post that article when I can find it. Lots of interesting voodoo

Intravenous lidocaine is very effective for chronic and acute pain, so I have considered it, but oral ambroxol seems so much easier.

What amazes me is that these sodium channels blockers have both an immediate action on pain, and a delayed action on inflammation, and I need both....

I am also interested on Ambroxol because it was suggested it may have an effect on tinnitus.

The thing is it may not be a good association with T3, so I will see if I can stop it while on Ambroxol.

https://academic.oup.com/bja/article/109/2/200/253255

 

[Inara]

Intravenous lidocaine is very effective for chronic and acute pain, so I have considered it, but oral ambroxol seems so much easier.

This reminds me - there are lidocain patches. They are usually used for neuropathic pain following varicella zoster infection, but also for other neuropathic pain. I find them very helpful, and you don't have the usual side effects.

 

[Iritu1021]

This looks like another promising way to hack into calcium modulation since there are quite a few ligands already out there for this receptor:

The sigma-1 receptor (σ1R), one of two sigma receptor subtypes, is a chaperone protein at the endoplasmic reticulum(ER) that modulates calcium signaling through the IP3 receptor.

Interestingly, pregnenolone, progesterone and DHEA are all ligands for this receptor @drob31

@BadBadBear @pattismith @Inara