Deeply disturbing information on combining longevity supplements

[John_Hemming]

No need to directly engage in the debate while others do it for you.

They can only do this if they answer questions about what happens when he does what he does. I accept this could be done by one of his specialists, but that is not happening.

Hence there is not really that much anyone can learn from what he is doing.

[DeStrider]

Therefore if he does have a representative here, it would be best if they engaged formally as his representative. I agree. Let’s all work together towards a common goal.

But in retrospect, he probably doesn’t.

[Ulf]

I don´t think he or his doctors will have a clue. The interactions between his drugs are uncountable, the pathways are many and what affects what will be impossible to say.

Below are examples of mTOR effects of one of the supps I looked at. The first three studies show mTOR activation, the last inhibition. A small example of the intricacies.

Taurine suppresses ROS-dependent autophagy via activating Akt/mTOR signaling pathway in calcium oxalate crystals-induced renal tubular epithelial cell injury - PMC (nih.gov)

Taurine attenuates methamphetamine-induced autophagy and apoptosis in PC12 cells through mTOR signaling pathway - ScienceDirect “These results indicated that taurine inhibits METH-induced autophagic process through activating mTOR rather than Erk signaling”.

My note: “Inhibiting AMPK/mTOR signalling pathway” actually means activation of mTOR, since mTOR is primarily inhibited by the activation mechanism of AMP-activated protein kinase (AMPK) and activated when AMPK is inhibited.

Taurine attenuates OTA-promoted PCV2 replication through blocking ROS-dependent autophagy via inhibiting AMPK/mTOR signaling pathway - PubMed (nih.gov) From the text “Furthermore, taurine supplementation inhibited 5’AMP-activated protein kinase (AMPK) and activated mammalian target of rapamycin (mTOR)”.

Taurine Reprograms Mammary-Gland Metabolism and Alleviates Inflammation Induced by Streptococcus uberis in Mice - PubMed (nih.gov) “These outcomes depend on taurine-mediated activation of the AMPK-mTOR pathway, which inhibits the over activation of inflammatory responses and alleviates cellular damage” “Taurine effectively inhibits overall energy metabolism by boosting AMPK levels reducing the activity of the mTOR pathway”. “Immunoblot analyses showed that taurine pretreatment significantly increased AMPK phosphorylation levels and subsequently downregulated mTOR”

[Acarb]

Has anyone considered Wormbot to test drug combinations? What is the cost and is it cost-effective?

- Healthy Aging and Longevity Research Institute

Using a web-based interface, the WormBot enables fully autonomous data collection from hundreds of parallel experiments simultaneously. The automated lifespan scoring features of the WormBot software are able to differentiate between pharmacological, environmental, and genetic treatments known to increase and decrease lifespan, and are comparable to human collected and annotated lifespan and behavioral data, but at substantially reduced time and effort. In addition to quantifying mortality under standard conditions, the WormBot can easily resolve survival differences under toxic conditions such as hydrogen cyanide exposure and can be used to quantify healthspan metrics such as motility and response to stress.

The following services are available to all users. Additional services may also be available and users are encouraged to contact the Core Director to discuss specific experimental details. Typical costs are provided below for initial budget preparation and may be used for Pilot Project submissions, but in all cases final costs will be determined following consultation with Core personnel.

Caenorhabditis elegans WormBot Assay capabilities

1. WormBot assays. The Core will perform lifespan and healthspan assessments using the WormBot platform [1]. Cost: Estimated costs are $500 per plate. Each plate can accommodate 12 experiments of 20-30 animals per experiment.
2. Healthspan measures.The Core will assay a variety of age-associated healthspan measures, including: motility, resistance to polyglutamine or amyloid beta toxicity, and reproductive capacity (brood size). Additional assays, including fluorescence reporter assays, may be available upon request. Cost:Cost will vary on a case-by-case basis, depending on services requested.
3. Resistance/sensitivity to environmental or chemical stresses. The core will perform survival assays in the presence of different environmental or chemical stressors upon request. Cost:Cost will vary on a case-by-case basis, depending on services requested.

[John_Hemming]

Wormbot is an interesting idea, but is still flailing around without a mechanistic hypothesis.

[KarlT]

Wasn’t widespread, maybe somewhat random, testing what Dr K. Talked about doing? His million molecules movement to quickly find more substances to study.

[Olafurpall]

@Ulf

He also told me that he believes all this is true even if different pathways are used to reach the same target. He seemed to agree that if supps don´t share the same target, the risk should be less or nullified. But this may be of scant consolation since the more I look the more everything seems to be related:

I agree with the notion that this is only a significant concern for supplements that share the same target. I think for most supplements, they don’t share the same main targets. This is specially true when it comes to vitamins and minerals. Supplements are more likely to overlap on smaller indirect targets.

@Ulf

Our talk departed from the mTOR signalling pathway and my use of Ca AKG, an mTOR inhibitor. As I research my supp stack, I realize that “All roads lead to mTOR”. In the crosstalk between signalling pathways, almost all my supps interact with the mTOR pathway: Besides AKG, they are taurine, curcumin, CoQ10, quercetin, carnosine, zinc, melatonin, tadalafil and reishi, They all inhibit mTOR except one that mainly activates mTOR.

I disagree that all roads lead to mTOR here. Among those you mentioned above, none of them influence mTOR to any significant degree in vivo. I wouldn’t be concerned about them interfering with the potential benefits of rapamycin.

[Joseph_Lavelle]

I’m not sure what you are asking. The original idea was to use him as a “canary in the coal mine”. I am saying that I will not rely on that approach.

[scta123]

I was trying to point out that while criticizing his approach we should look at our approach. It is mostly the same on one level or another.

[Joseph_Lavelle]

Then we agree. I think his approach is crazy. I think my approach is crazy, just less so. I am backing down now.

1. Zero supplements on rapa days: 0, 5 & 6
2. I’m arranging interviews right now with people in the longevity biz to collect thoughts on establishing guardrails for staying within a safety zone.
3. A part of that guardrail system will be, I believe, a limit on “risky” supplements (tbd) that will require me to drop 1 to pick-up 1. No slippery slope of wishful thinking. I expect to have to drop down from where I am now before maintaining.

I’m open to ideas. How else could I establish a safety zone for my longevity program?

Also, let me know if you have questions for my interviews on this topic. Guest names to be announced as arrangements are made.

Thanks

[Joseph_Lavelle]

As long as the cost per look is low, I think it’s a good idea to look in places where there is no reason to believe anything good would happen. That is how we will find new big ideas. Once they find something unexpected, they can chase down the mechanism. I think Kaeberlein’s thesis is that today’s science knows too little to focus on what we expect to find.

[John_Hemming]

I think it is difficult because of the timing issues and combinatory issues, but I understand the idea.

Citrate has actually been tried with fruit flies. It found a life extension, but it was not a subtle detailed experiment.

The balance between acetyl-CoA issues and mitochondrial efficiency will vary from species to species.

[Ulf]

Joseph: Great if you can make interviews, and even get guidance on risks of cancelling-out effects between specific substances. I mentioned some I am/was taking, but you will have your own list.

[Phil_Van_Treuren]

@Joseph_Lavelle

I’m arranging interviews right now with people in the longevity biz

This is awesome, very proactive approach!

[desertshores]

You may have mentioned this before, but how much citrate supplementation is effective?

[Joseph_Lavelle]

Can anyone get this?

Combinatorial interventions in aging

https://www.nature.com/articles/s43587-023-00489-9

[John_Hemming]

I think you will see effects at 4g per day. Currently I alternate between 20 and 40g per day taken in 5g quantities at Least 30 mins apart.

Question as to what to do with my protocol

Warning: There may be unidentified risks in trying parts of this protocol. Anyone who tries part of the protocol needs to monitor their biomarkers and if any harm is being caused stop using it. No medical advice is being given people need to do their own calculations and assessments and take medical advice from a healthcare professional where appropriate. People take their own decisions at their own risk. Get a baseline. Before doing anything it really is worth having a baseline. This really has to include high sensitivity CRP. There are costs to testing and the costs of testing probably exceed the costs of the protocol. A baseline test before starting anything is really helpful. The principle The principle behind this protocol is that if you improve how cells work then you improve how the body works. Cells function by generating energy and producing proteins. If a cell is unable to produce the full range of proteins then it will not function properly. The theory underp…

[anon16510610]

Move along, nothing to see here.

[Ulf]

Olafurpall:

I would be very relieved if most roads do not lead to mTOR so I can continue taking newly purchased highest quality supps!

What I found on those substances made me think that their influence on mTOR could be significant. However, the links are mainly in vitro and I take it you mean that those can be disregarded, if you have found evidence of no or only limited in vivo effects that contradict those.

On a few I believe there is in vivo evidence of links (zinc, taurine, tadalafil and reishi). But perhaps the links are weak?.

It is very long but for FWIW I post the links I found between the substances I was (am) taking and mTOR (taurine I already posted on). Some such as Quercetin lit up like a Christmas tree when I searched, others e.g. CoQ10 I had to hunt before finding the links. Vitamin K2 was about the only substance that I didn´t find a link on.

CURCUMIN

As the first study shows, mTOR appears to be a key target of curcumin but there may be other central targets.

From Hitting the Golden TORget: Curcumin’s Effects on mTOR Signaling | Bentham Science

“mTOR has emerged as an exciting and novel molecular target for curcumin, particularly in cancer cell lines. It appears that curcumin inhibits both mTORC1 and mTORC2”

“These studies also suggest that curcumin possesses amazing molecular versatility as evidenced by the large number of proposed drug receptors for the compound, including plasma membrane-bound receptors, proteases, transporters, apoptotic factors, kinases, transcription factors, and adhesion molecules [2, 12]. While it remains a possibility that curcumin is in fact interacting with this seemingly large range of molecules, a more plausible explanation is that curcumin predominately targets only a few key cell regulators, and these actions spill over to affect many different pathways, factors, and processes within the cell. Identifying these key cellular regulators, however, remains challenging due to the seemingly diverse molecular promiscuity of the compound. Recently, the mammalian target of rapamycin (mTOR) has been identified as a novel molecular target of curcumin, which may in fact represent one of these central targets due to the fact that mTOR stands at the center of numerous key cellular processes (cell growth/proliferation, survival and motility), almost all of which are affected to some degree by curcumin”.

From other studies “Curcumin has been suggested to regulate several cellular signaling pathways including mTOR signaling and functions as a new class of mTOR inhibitor”

“ Curcumin is termed as a multifunctional targeting therapy drug that regulates the mTOR signaling pathway in the treatment of numerous diseases”.

QUERCETIN

Frontiers | Quercetin Can Improve Spinal Cord Injury by Regulating the mTOR Signaling Pathway (frontiersin.org)

Inhibition of mTOR signaling by quercetin in cancer treatment and prevention - PubMed (nih.gov)

Quercetin induces apoptosis and autophagy in primary effusion lymphoma cells by inhibiting PI3K/AKT/mTOR and STAT3 signaling pathways - PubMed (nih.gov)

CoQ10

Coenzyme Q10 Ameliorates Pancreatic Fibrosis via the ROS-Triggered mTOR Signaling Pathway - PubMed (nih.gov) “In primary PSCs, expression levels of p-PI3K, p-AKT, and p-mTOR were upregulated with CoQ10. A rescue experiment using specific inhibitors of the PI3K-AKT-mTOR pathway demonstrated that the PI3K-AKT-mTOR signaling pathway was the underlying mechanism by which CoQ10 ameliorated fibrosis”.

Coenzyme Q10 Inhibits the Aging of Mesenchymal Stem Cells Induced by D-Galactose through Akt/mTOR Signaling - PMC (nih.gov) “These data indicate that the Akt/mTOR signaling plays a critical role in MSCs senescence inhibited by CoQ10”. “The results suggested that CoQ10 could inhibit D-gal-activated Akt/mTOR signaling in MSCs”……” Therefore, we hypothesized that Akt/mTOR signaling inactivation might be the mechanism by which CoQ10 inhibited MSC aging induced by D-gal. Our result showed that the expression of phosphorylated Akt and mTOR could be deceased by CoQ10. Finally, after overexpression of constitutively active Akt (CA-Akt), the number of SA-β -gal–positive cells was increased and the level of p53, p21, and p16 was also elevated in the CoQ10 treatment group; these results hint that the Akt/mTOR signaling may be the main mediator of MSC aging regulated by CoQ10”.

ZINC

Zinc promotes autophagy and inhibits apoptosis through AMPK/mTOR signaling pathway after spinal cord injury - PubMed (nih.gov)

The mTOR target is primarily inhibited by the activation mechanism of AMP-activated protein kinase (AMPK) [1]. Yun et al. found that activation of phospho-mTOR inhibited autophagy, whereas the inhibition of phospho-mTOR induced autophagy [29]. Cao et al. found that activation of autophagy increased AMPK phosphorylation with a reduction in phosphorylation of the S6 kinase P70 subtype (P70S6K) [4].

It was shown that zinc significantly induced the level of Beclin1 and LC3B by activating adenosine 5′-monophosphate (AMP)-activated protein kinase (AMPK) signaling pathway.

Moreover, recent studies have demonstrated that zinc is the regulation of autophagy.

Zinc enhances intestinal epithelial barrier function through the PI3K/AKT/mTOR signaling pathway in Caco-2 cells - PubMed (nih.gov)

Effect of supplemental dietary zinc on the mammalian target of rapamycin (mTOR) signaling pathway in skeletal muscle and liver from post-absorptive mice - PubMed (nih.gov)

MELATONIN

Melatonin attenuates vascular calcification by activating autophagy via an AMPK/mTOR/ULK1 signaling pathway - PubMed (nih.gov)

Melatonin significantly increased expression of p-AMPK and p-ULK1, and decreased mTOR expression. Treatment with compound C (an inhibitor of AMPK) or MHY1485 (an agonist of mTOR) ablated the observed benefits of melatonin treatment. Melatonin protects VSMCs against calcification by activating autophagy via the AMPK/mTOR/ULK1 pathway.

Melatonin inhibits proliferation, migration, and invasion by inducing ROS-mediated apoptosis via suppression of the PI3K/Akt/mTOR signaling pathway in gallbladder cancer cells - PubMed (nih.gov)

Melatonin induces autophagy via an mTOR-dependent pathway and enhances clearance of mutant-TGFBIp - PubMed (nih.gov)

The last sentence shows a nice additive effect with rapa: “Our results show that melatonin activates autophagy in both wild-type (WT) and GCD2-homozygous (HO) corneal fibroblast cell lines via the mammalian target of rapamycin (mTOR)-dependent pathway. Melatonin treatment also led to increased levels of beclin 1, which is involved in autophagosome formation and maturation. Furthermore, melatonin significantly reduced the amounts of mutant- and WT-TGFBIp. Treatment with melatonin counteracted the autophagy-inhibitory effects of bafilomycin A1, a potent inhibitor of autophagic flux, demonstrating that melatonin enhances activation of autophagy and increases degradation of TGFBIp. Cotreatment with melatonin and rapamycin, an autophagy inducer, had an additive effect on mutant-TGFBIp clearance compared to treatment with either drug alone**”.**

TADALAFIL

Tadalafil and bergapten mitigate streptozotocin-induced sporadic Alzheimer’s disease in mice via modulating neuroinflammation, PI3K/Akt, Wnt/β-catenin, AMPK/mTOR signaling pathways - ScienceDirect

“ Furthermore, TAD tadalafil and BG boosted hippocampal levels of cGMP, PKG, Wnt3a, and AMPK and reduced expression of β-catenin and mTOR by 74% and 51%, respectively. TAD and BG also halted neuroinflammation by reducing IL-23 and IL-27 levels, as well as protein expression of NF-κB by 62% & 61%, respectively. In conclusion, this study offers novel insights on the neuroprotective effects of TAD or BG in the management of SAD as evidenced by improved cognitive function and histological architecture. This could be attributed to modulation of the crosstalk among PI3K/Akt/GSK-3 β , PP2A, mTOR/autophagy, cGMP/PKG, and Wnt/ β -catenin signaling cascades and mitigation of neuroinflammation”.

Tadalafil Treatment of Mice with Fetal Growth Restriction and Preeclampsia Improves Placental mTOR Signaling - PubMed (nih.gov)

The expression of p-mTOR was significantly decreased in mice with FGR on 13 days post coitum (d.p.c.) but recovered to the same level as that of the control on 17 d.p.c. following tadalafil treatment. The results were similar for 4E-binding protein 1 (4E-BP1) and S6 ribosomal (S6R) protein, which act downstream in the mTOR signaling pathway. We demonstrate that the tadalafil treatment of FGR in mice improved placental mTOR signaling to facilitate fetal growth.

Tadalafil Treatment Ameliorates Hypoxia and Alters Placental Expression of Proteins Downstream of mTOR Signaling in Fetal Growth Restriction - PubMed (nih.gov)

CARNOSINE

Carnosine attenuates vascular smooth muscle cells calcification through mTOR signaling pathway - PubMed (nih.gov)

Carnosine Inhibits the Proliferation of Human Gastric Carcinoma Cells by Retarding Akt/mTOR/p70S6K Signaling - PMC (nih.gov)

Carnosine inhibits glioblastoma growth independent from PI3K/Akt/mTOR signaling - PubMed (nih.gov)

REISHI

An in vivo study on mice injected with inflammatory breast cancer (IBC) and treated with the commercial extract ReishiMax GLpTM (carpophore and cracked spores) highlighted a selective action on gene and protein expression, with smaller tumor size and weight and reduced expression of E-cadherin, mammalian target of rapamycin (mTOR),

VITAMIN D

Vitamin D - PMC (nih.gov)

“We have shown that 1,25(OH)2D is able to regulate the mammalian/mechanistic target of rapamycin (mTOR) signaling pathway by stimulating expression of DNA damage-inducible transcript 4 (DDIT4), also known as regulated in development and DNA damage response 1 (REDD1), a potent suppressor of mTOR activity.2 Given the role of mTOR as a “master regulator” of cell function,3 it seems likely that DDIT4-mediated inhibition of this pathway will also play a pivotal role in mediating cellular responses to 1,25(OH)2D, as well as provide new strategies for its use in disease therapy”.

“Results demonstrated that the addition of Vitamin D to insulin and leucine significantly enhanced the activity of the mTOR pathway and protein synthesis. The authors conclude that Vitamin D has the potential to directly alter protein synthesis in muscle cells”

BETA GLUCAN

I only find glimpses of possible direct action from beta glucan on mTor:

mTOR/HIF1α-mediated aerobic glycolysis as metabolic basis for trained immunity - PMC (nih.gov) The gene expressing mTOR and the glycolytic genes that are targets of the transcription factor HIF1 α were also enhanced by β-glucan

But the indirect links appear clear, from reading both the following link extracts

Biomedical aspects of beta-glucan on glucose metabolism and its role on primary gene PIK3R1 - ScienceDirect

β- glucan plays a significant role in regulating the PI3K/Akt biochemical pathway via IRS1, PI3K-p85, and phosphorylated Akt, which increases insulin sensitivity and reduces the associated risks.

From PI3K/AKT pathway as a key link modulates the multidrug resistance of cancers | Cell Death & Disease (nature.com)

“PI3K binds to the PH domain of AKT and induces conformational changes and the subsequent phosphorylation of AKT, and activated AKT moves from the cytoplasm to the cell membrane. This cascade directly or indirectly activates its downstream molecular proteins, such as NF-κB and mTOR21”.”

[John_Hemming]

My view is that mTOR is important because it is a route towards autophagy/mitophagy which is a route towards improved mitochondrial efficiency.

COQ10 and Vitamin K2 (menaquninone-7) are both routes towards improved mitochondrial efficiency whether or not they have any effect on mTOR. (Melatonin helps as well).

I think HDAC inhibitors have a separate important role that includes: Quercetin, Curcumin, etc

Vitamin D is a route toward the VDR being active and that has an important cellular function.