Year: 2025

Plasmapheresis doesn’t reduce biological age

gettingwell4 4-5 stars Advanced knowledge, Epigenetics ,

A 2025 clinical trial investigated effects of plasmapheresis as measured with epigenetic clocks:

“This study aimed to assess whether plasmapheresis without volume replacement with young plasma or albumin affects epigenetic age and other biomarkers in healthy adults. No significant epigenetic rejuvenation was observed based on epigenetic clock measurements. Instead, plasmapheresis was associated with increases in DNAmGrimAge, the Hannum clock, and the Dunedin Pace of Aging.

  1. The relatively small sample size of 34 finishing participants comprising of first-time plasma donors limits the statistical power and generalizability of our findings.
  2. Our cohort was restricted to individuals aged 40 to 60 years in accordance with Czech regulatory guidelines, which, although intentional to focus on an older population where rejuvenating effects might be most apparent, constrains evaluation of age-related differences across a broader demographic.
  3. The 18-week duration of the study, while sufficient to detect rapid alterations in key biomarkers under an intensive plasmapheresis protocol, may not fully capture the long-term implications of these changes.
  4. Due to our trial taking place during spring and summer months, we cannot fully separate the effects of increased sunlight exposure, outdoor physical activity, and dietary changes from the observed rises in Vitamin D and concurrent shifts in DNAm-based aging metrics. We did not collect objective measures of activity or diet, so these factors remain potential confounders.

The protocol of donating plasma every two weeks, although deemed safe by many countries around the world, is not yet well researched and cannot therefore be marked as benefiting to the donor right now. Further refinement to balance clearance of pro-aging factors with maintenance of systemic homeostasis is needed.”

https://www.nature.com/articles/s41598-025-05396-0 “Human clinical trial of plasmapheresis effects on biomarkers of aging (efficacy and safety trial)”

Broccoli sprouts and your dog

gettingwell4 2-3 stars Required further work, Stress

This 2025 opinion paper compared nine broccoli sprouts supplements for dogs:

“Broccoli sprouts are key elements of 9 dietary supplements for dogs. Feeding directions of 6 products correspond with consumption of dry food containing 0.5 to 29 g dried broccoli sprouts/kg. Seven supplements claim to supply sulforaphane and to possess anti-inflammatory and/or anti-cancer effects.

Directions for use of a sulforaphane-producing supplement read as follows: ‘One chewable tablet daily for dogs of all sizes, six months and older. Tablets should be given on an empty stomach at least two hours after a meal or one hour before a meal.’

There was no information found on feeding studies in dogs, addressing the impact of broccoli sprouts on health. In mice, dietary, whole-broccoli sprouts counteracted development of mammary and prostate cancer. Weights of dried broccoli sprouts in these mouse studies were 150 and 260 g/kg dry food, much higher levels than equivalents of feeding instructions for dog supplements. Species contrast and high dose blunt extrapolation of results to dogs.

The Veterinary Clinical Trials Registry of the American Veterinary Medical Association has announced that recruiting has finished for a study entitled “Sulforaphane supplementation in canine lymphoma and evaluation of epigenetic proteomic profiles”. https://veterinaryclinicaltrials.org/study/VCT17004227

https://www.researchgate.net/publication/393656647_Beynen_AC_2025_Broccoli_sprouts_in_dog_nutrition “Beynen AC, 2025. Broccoli sprouts in dog nutrition” (registration required)

The author’s use of ResearchGate is mainly to publish opinion pieces on pet animal nutrition. This doesn’t require high fees of regular journals, but also bypasses peer review.

I appreciate comparisons to rodent studies, which often intentionally overdose, and so have no relevance to humans and other mammals. His 2025 pet nutrition papers include broccoli, glyphosphate, zinc, copper, and PFAS subjects.

Betaine as an exercise mimetic

gettingwell4 5+ stars Premium, Cortisol, Epigenetics, Immune system, Memory, Neurochemicals, Stress , ,

A 2025 human study investigated effects of long-term exercise:

“Exercise has well-established health benefits, yet its molecular underpinnings remain incompletely understood. We conducted an integrated multi-omics analysis to compare effects of acute vs. long-term exercise in healthy males.

Acute exercise induced transient responses, whereas repeated exercise triggered adaptive changes, notably reducing cellular senescence and inflammation and enhancing betaine metabolism. Exercise-driven betaine enrichment, partly mediated by renal biosynthesis, exerts geroprotective effects and rescues age-related health decline in mice.

Betaine binds to and inhibits TANK-binding kinase 1 (TBK1), retarding the kinetics of aging.

Betaine effectively alleviated senescence phenotypes by reduced senescence-associated β-galactosidase (SA-β-Gal)-positive cells, decreased p21 expression, lowered DNA damage indicator γ-H2A.X, and elevated heterochromatin mark H3K9me3. Betaine treatment also enhanced cellular antioxidant capacity, as evidenced by increased NRF2 phosphorylation and reduced ROS accumulation.

These findings systematically elucidate the molecular benefits of exercise, and position betaine as an exercise mimetic for healthy aging.”

https://doi.org/10.1016/j.cell.2025.06.001 “Systematic profiling reveals betaine as an exercise mimetic for geroprotection” (not freely available) Thanks to Dr. Weimin Ci for providing a copy.

Taurine and mitochondrial health

gettingwell4 4-5 stars Advanced knowledge, Immune system, Stress

A 2025 review subject was taurine’s beneficial effects on mitochondria:

“Taurine has multiple and complex functions in protecting mitochondria against oxidative-nitrosative stress. We introduce a novel potential role for taurine in protecting from deuterium (heavy hydrogen) toxicity. This can be of crucial impact to either normal or cancer cells that have highly different mitochondrial redox status.

Deuterium is an isotope of hydrogen with a neutron as well as a proton, making it about twice as heavy as hydrogen. We first explain the important role that the gut microbiome and gut sulfomucin barrier play in deuterium management. We describe synergistic effects of taurine in the gut to protect against deleterious accumulation of deuterium in mitochondria, which disrupts ATP synthesis by ATPase pumps.

Taurine’s derivatives, N-chlorotaurine (NCT) and N-bromotaurine (NBrT), produced through spontaneous reaction of taurine with hypochlorite and hypobromite, have fascinating regulatory roles to protect from oxidative stress and beyond. We describe how taurine could potentially alleviate deuterium stress, primarily through metabolic collaboration among various gut microflora to produce deuterium depleted nutrients and deuterium depleted water (DDW), and in this way protect against leaky gut barrier, inflammatory bowel disease, and colon cancer.

Taurine cannot be metabolized by human cells, but gut microbes are able to break it down to release sulfite, which then gets oxidized to sulfate anions that become available to support synthesis of sulfomucins. Taurine protects against many diseases linked to mitochondrial defects, such as aging, metabolic syndrome, cancer, cardiovascular diseases and neurological disorders.

We present a novel view that gut microbes play an essential role in providing deuterium depleted (deupleted) nutrients, especially, butyrate, to the host colonocytes forming the gut barrier. We propose that sulfomucins synthesized by goblet cells not only protect the barrier from pathogens, but also trap and sequester deuterium, thus reducing mitochondrial deuterium levels, resulting in improved mitochondrial health.

Due to taurine, redox buffer glutathione (GSH) further stabilizes the membrane potential. GSH not only reduces radical oxygen species (ROS) during oxidative stress, but it also assists in production of deupleted water in mitochondria.

Spontaneous oxidation of two GSH molecules to produce GSSG in the presence of hydrogen peroxide yields two molecules of DDW. Just as for glutathione, bilirubin can produce DDW indefinitely through chronic recycling between bilirubin and biliverdin, capturing a deupleted proton in NADPH to produce a DDW molecule in each cycle.

A novelty that arises from this investigation is introduction of the role that deuterium plays in mitochondrial disease, and ways in which taurine may facilitate maintenance of low deuterium in mitochondrial ATPase pumps. Excess deuterium causes a stutter in the pumps, which leads to inefficiencies in ATP production and an increase in ROS.”

https://pmc.ncbi.nlm.nih.gov/articles/PMC11717795/ “Taurine prevents mitochondrial dysfunction and protects mitochondria from reactive oxygen species and deuterium toxicity”

Stay away from NAC

gettingwell4 4-5 stars Advanced knowledge, Cortisol, Hypothalamus, Immune system, Limbic system, Neurochemicals, Stress

A 2025 rodent study added several reasons to avoid non-emergency use of N-acetylcysteine:

“We previously showed that antioxidants induced an impairment of negative feedback of the hypothalamus-pituitary-adrenal (HPA) axis in rats, in parallel to a down-regulation of glucocorticoid receptor (GR) and nuclear factor erythroid 2-related factor 2 (Nrf2) expression in the pituitary gland. This study evaluated the role of the Nrf2-heme-oxygenase-1 (HO-1) pathway on impairment of negative feedback of the HPA axis induced by N-acetylcysteine (NAC).

Male Swiss-Webster mice were orally supplemented with NAC for 5 consecutive days. The Nrf2-HO-1 pathway activator cobalt protoporphyrin IX (CoPPIX) was injected intraperitoneally on days 2 and 5 after starting NAC supplementation.

NAC reduced expression of Nrf2 in the pituitary of mice. NAC induced adrenal enlargement and hypercorticoidism, along with a decrease in GRα expression and an increase of GRβ expression in the pituitary gland.

We observed that dietary supplementation with NAC ( Figure 4A ) significantly increased plasma corticosterone levels in mice 24h ( Figure 4B ) as well as 15 days ( Figure 4C ) after the last administration of the antioxidant with the same magnitude of response (3.5-fold and 3.4-fold, respectively).

Chronic activation of the HPA axis can have damaging effects on immune, cardiovascular, metabolic, and neural functions, increasing the risk of immune system dysfunction, mood disorders, and metabolic and cardiovascular diseases. To prevent these deleterious effects of chronic hypercortisolism, HPA axis function is controlled by a glucocorticoid-dependent negative feedback system that is essential for ending the stress response.

These findings showed that NAC reduced Nrf2-HO-1 pathway activation in the pituitary gland, in a mechanism probably related to a local downregulation of GRα and an up-regulation of GRβ, leading to a failure of negative feedback of the HPA axis and consequently to the hyperactivity of this neuroendocrine axis.”

https://pmc.ncbi.nlm.nih.gov/articles/PMC11827418/ “Activation of the Nrf2/HO-1 pathway restores N-acetylcysteine-induced impairment of the hypothalamus-pituitary-adrenal axis negative feedback by up-regulating GRα expression and down-regulating GRβ expression into pituitary glands”

A human equivalent to this study’s NAC dose is (150 mg x .081) x 70 kg = 851 mg. Human supplements are sold in 600 mg and 1000 mg doses.

Grok 3 replied to a question: Human equivalent time to 15 days in male Swiss-Webster mice aged between 4 and 6 weeks? by stating: “15 days in male Swiss-Webster mice aged 4 to 6 weeks corresponds to approximately 4.1 human years, advancing their equivalent human age from about 10–12 years to roughly 14–16 years.” Four+ years seems like a long time for NAC to steadily and continuously affect humans’ HPA axes per the above graphic. What do you think?

Previously mentioned reasons to avoid daily use of NAC were in the lower part of A good activity for bad weather days.

Eat broccoli sprouts for your HIIT, Part 2

gettingwell4 2-3 stars Required further work, Epigenetics, Stress

A 2025 human study followed up Eat broccoli sprouts for your high intensity interval training using red kale sprouts:

“Glucosinolate-rich broccoli sprouts combined with intense exercise training for 7 days have been shown to reduce blood lactate concentrations during exercise, attenuate hypoglycemic events, improve physical performance, and reduce markers of oxidative stress. This study aimed to investigate the acute, dose-dependent effects of glucosinolate-rich red kale sprouts (GRS) on blood lactate and blood glucose following the ingestion of three different doses.

Fifteen healthy participants [11 females, 4 males] consumed 37.5 g or 75 g of GRS or an isocaloric placebo blended into a beverage on three separate occasions. The participants cycled on an ergometer at three submaximal work rates before and three hours after ingestion.

Intake of glucosinolate-rich sprouts acutely decreased blood lactate levels during submaximal cycling and increased blood glucose levels at rest. The largest reduction in blood lactate was observed at the 37.5 g dose compared to placebo.

To identify the dose of GRS that results in the lowest blood lactate concentration during submaximal exercise, we applied a quadratic modeling approach. The optimal dose for minimizing lactate accumulation was calculated as 44 g of GRS.

In our previous study, we found a tendency towards a lower respiratory exchange ratio after one week of supplementation. Moreover, studies have demonstrated that mitochondrial oxidation of long-chain and short-chain fatty acids is depressed in the absence of Nrf2, and accelerated when Nrf2 is constitutively active.

We observed a reduction in myeloperoxidase levels approximately three hours after GRS intake, suggesting a decrease in oxidative stress. This finding indicates that the adaptive compensatory system may act rapidly, likely within just a few hours of GRS consumption.

A limitation is that we did not assess whether the lower lactate levels translated into improved performance. Theoretically, if the reduction in lactate results from the activation of pyruvate dehydrogenase, it could enhance performance by channeling more pyruvate into mitochondria for efficient oxidation, reducing reliance on glycolysis, and thereby sparing muscle glycogen. Alternatively, if the lower lactate levels are due to increased activity of the hepatic Cori cycle, lactate could be more rapidly converted to glucose, possibly supporting glycogen resynthesis or maintaining blood glucose levels during exercise. Both mechanisms could potentially contribute to improved performance.

Acute intake of small doses of GRS followed by submaximal ergometer cycling results in changes in lactate and glucose metabolism that could be beneficial for exercise performance.”

https://www.biorxiv.org/content/10.1101/2025.04.15.648889v1.full “A glucosinolate-rich beverage lowers blood lactate concentrations during submaximal exercise”

These researchers chose red kale sprouts of undisclosed age over the predecessor study’s broccoli raab five-day old sprouts, and two other undisclosed cruciferous vegetable sprouts.

This study is in its preprint phase. Items that could be clarified before publishing in final form include:

  • In the Abstract section, reference findings to red kale sprouts rather than broccoli sprouts;
  • Characterize the lactate U-shaped dose-response curve as hormesis; and
  • Reference other hormesis findings for context.

Sulforaphane as a senotherapy

gettingwell4 4-5 stars Advanced knowledge, Epigenetics, Immune system, Stress

A 2025 rodent study investigated cardioprotective effects of dasatinib / quercetin (a senolytic combination) and sulforaphane (senomorphic):

“Senolytics are molecules that selectively eliminate senescent cells (SCs). Senomorphics are drugs that suppress or mitigate the pro-inflammatory effects of the senescence-associated secretory phenotype (SASP) without killing SCs. Senomorphics decrease the number of SCs by diminishing the paracrine effect of senescence induction in neighboring cells, and by enhancing elimination of SCs by the immune system, which reduces their harmful effects.

We used middle-aged female rats fed a hypercaloric diet (HD) from 21 days to 15 months of age. Under our experimental conditions, rats exhibited cardiac hypertrophy and fibrosis, accumulation of senescent cells, changes in mitochondrial morphology, and oxidative stress. Rats were treated for 2 months with senolytic (dasatinib + quercetin, DQ) or senomorphic (sulforaphane, SFN) agents.

A novel aspect of recent research has been the crosstalk between organelles, particularly between the mitochondria and endoplasmic reticulum (ER), through specialized contact sites (MERCSs). However, there is still no consensus on the optimal distance between MERCSs and their positive or negative effects on disease progression.

HD rats showed cardiac improvement after both treatments. Although both strategies improved cardiomyocyte size and cardiac fibrosis, only DQ decreased LDH levels, whereas SFN positively affected cardiac damage proteins.

In general, no changes in structure or damage-associated enzymes were observed in control rats treated with DQ or SFN, indicating that senotherapies do not promote adverse effects on the heart, reinforcing the concept that they are safe for application in the clinical field. Data suggest a possible link mechanism between Nrf2 activation and MERCSs preservation, activated by SFN rather than by the DQ combination, which allowed cardiac structure maintenance in HD rats decreasing harmful effects of senescent cells.”

https://www.sciencedirect.com/science/article/abs/pii/S0955286325000865 “Cardioprotective effect of senotherapy in chronically obese middle-aged female rats may be mediated by a MERCSs/Nrf2 interaction” (not freely available) Thanks to Dr. Alejandro Silva for providing a copy.

Eat broccoli sprouts for your eyes, Part 3

gettingwell4 4-5 stars Advanced knowledge, Cerebellum, Epigenetics, Hippocampus, Limbic system, Lower brain, Stress , , , ,

Two 2025 papers cited Precondition your defenses with broccoli sprouts, starting with a review of age-related macular degeneration:

“AMD progression from intermediate to late AMD leads to a point of irreversible retinal pigmented epithelium (RPE) degeneration where treatment becomes worthless. Treating patients at the early/intermediate stages presents a better therapeutic window opportunity for AMD as the disease could potentially be prevented or slowed down.

Strong evidence points to RPE dysfunction at these stages, mainly through redox imbalance and lysosomal dysfunction in RPE oxidative injury. Restoring oxidative balance and lysosomal function may act as preventive and therapeutic measures against RPE dysfunction and degeneration.

Due to interaction with KEAP1, NRF2 is a ubiquitously expressed protein with a high turnover and half-life of about 20 minutes. Because the turnover of NRF2 is faster than KEAP1, newly synthesized NRF2 does not have free KEAP1 to bind and is translocated into the nucleus. Once in the nucleus, NRF2 dimerizes with sMAF and the complex binds to antioxidant response element (ARE) sequences, promoting the expression of ARE genes.

There is NRF2 involvement in most of the hallmarks of aging. Key transcriptional regulatory factors of related pathways, such as transcription factor EB (TFEB) and NRF2, may be targeted to restore homeostasis and/or prevent further RPE degeneration.”

https://www.mdpi.com/2076-3921/14/5/596 “Targeting Lysosomal Dysfunction and Oxidative Stress in Age-Related Macular Degeneration”

There were other informative tidbits throughout this review, such as:

  • “Anti-inflammatory effects of most electrophilic NRF2 activators are thought to be at least partly NRF2-independent, suggesting that these compounds lacking specificity may be advantageous for multitargeted pathologies.
  • TFEB can activate NRF2 under conditions devoid of oxidative stress.”

This paper also cited Bridging Nrf2 and autophagy when discussing the above graphic.

In this human cell and rodent study, several coauthors of the original 2020 study tested sulforaphane and TFEB interactions for ameliorating effects of a rare disease:

“Mutations in genes encoding lysosomal proteins could result in more than approximately 70 different lysosomal storage disorders. Niemann–Pick disease type C (NPC) is a rare lysosomal storage disorder caused by mutation in either NPC1 or NPC2 gene. Deficiency in NPC1 or NPC2 protein results in late endosomal/lysosomal accumulation of unesterified cholesterol.

Clinical symptoms of NPC include hepatosplenomegaly, progressive neurodegeneration, and central nervous system dysfunction, that is, seizure, motor impairment, and decline of intellectual function. So far there is no FDA-approved specific therapy for NPC.

Under stress conditions, that is, starvation or oxidative stress, TFEB is dephosphorylated and actively translocates into the nucleus, promoting expression of genes associated with lysosome and autophagy. TFEB overexpression or activation results in increased number of lysosomes, autophagy flux, and exocytosis.

Pharmacological activation of TFEB by sulforaphane (SFN), a previously identified TFEB agonist, significantly promoted cholesterol clearance in human and mouse NPC cells, while genetic inhibition (KO) of TFEB blocked SFN-induced cholesterol clearance. This clearance effect exerted by SFN was associated with upregulated lysosomal exocytosis and biogenesis. SFN treatment has no effect on the liver and spleen enlargement of Npc1 mice.

SFN is reportedly BBB-permeable, assuring a good candidate for efficient delivery to the brain, which is essential for targeting neurodegenerative phenotypes in neurological diseases including NPC. This is the first time that SFN was shown to directly activate TFEB in the brain.

Collectively, our results demonstrated that pharmacological activation of TFEB by a small-molecule agonist can mitigate NPC neuropathological symptoms in vivo. TFEB may be a putative target for NPC treatment, and manipulating lysosomal function via small-molecule TFEB agonists may have broad therapeutic potential for NPC.”

https://elifesciences.org/articles/103137 “Small-molecule activation of TFEB alleviates Niemann–Pick disease type C via promoting lysosomal exocytosis and biogenesis”

Interpreting results of a Vitamin K2 clinical trial

gettingwell4 4-5 stars Advanced knowledge

This 2025 paper analyzed clinical trial subjects’ protocol adherence:

“The VitaK-CAC (vitamin K-coronary artery calcification) trial is a double-blind, randomized, placebo-controlled trial in patients with pre-existent CAC who were treated for two years with either placebo or the vitamin K2-analogue menaquinone-7 (MK-7) [360 mcg daily]. The present analysis of the VitaK-CAC trial assesses degree of adherence to supplementation with MK-7 during the implementation and persistence phases.

We estimated adherence in three different ways: 1) by pill counts, 2) by measuring plasma levels of MK-7, and 3) by measuring plasma levels of dephosphorylated, uncarboxylated matrix Gla-protein (dp-ucMGP), a marker of functional bioactivity of vitamin K.

  1. We estimated tablet intake by counting the number of pills participants returned to the laboratory at each follow-up visit. When 20% or less of the prescribed pills were returned, this was considered adequate adherence. Due to inconsistent bottle returns by many participants, reliable pill counts could be obtained for only half of the patients. No difference in evolution of the CAC score was found between patients with an adherence below or above 80%, as determined by pill counts.
  2. In patients receiving MK-7, an increase in plasma levels above the upper interquartile range (IQR) observed in the placebo group was taken as evidence of supplementation adherence. When this occurred both at 12 months and at 24 months, this was taken as a robust indicator of good persistence. When patients with persistence throughout the study, as based on the MK-7 levels, were compared to those who were not, the former group showed an attenuated progression of their CAC score.
  3. We considered a fall in dp-ucMGP by more than 10% as indicative of a positive effect of MK-7 supplementation and, hence, as a proxy for good adherence. Levels of dp-ucMGP rose significantly in both groups, although to a lesser extent in the group with active treatment. If the reduced rise in dp-ucMGP is considered a proxy for adherence, this suggests approximately 60% adherence in the MK-7 group. In 21 of the 75 patients (28%), the rise in MK-7 was sufficiently great for levels of dp-ucMGP to decrease.

There was no difference in the response of MK-7 levels and in the primary outcome (CAC progression) between patients with or without sufficient adherence. We cannot exclude the possibility that variations in absorption or metabolism have contributed to, or may even be entirely responsible for, the observed swings in MK-7 plasma levels.

None of the three methods that we applied in our study is absolutely reliable to estimate adherence to supplementation with MK-7. Measurement of MK-7 levels provides the best information.”

https://www.explorationpub.com/Journals/em/Article/1001321 “Adherence to dietary supplementation with menaquinone-7, a vitamin K2 analogue”

A paper described the clinical trial itself:

https://journals.lww.com/jhypertension/fulltext/2025/05001/menaquinone_7_slows_down_progression_of_coronary.39.aspx “MENAQUINONE-7 SLOWS DOWN PROGRESSION OF CORONARY ARTERY CALCIFICATION. A RANDOMIZED, PLACEBO-CONTROLLED TRIAL” (not freely available, NCT01002157)

Always more questions:

1. The registration summary included:

In animal studies, active supplementation of Vitamin K2 caused regression of existing arterial calcification. The aim of this randomized, double-blind, placebo-controlled clinical trial is to investigate whether daily supplementation of Vitamin K2 (Menaquinone-7) to patients with established CAC will lead to a decreased progression-rate of CAC after 24 months of follow-up in comparison to placebo.

Is this trial’s stated goal to decrease CAC progression to the point of an unstated goal of reversing existing arterial calcification?

2. What is the degree of success of this clinical trial? I don’t have access to the clinical trial paper, although I’ve requested a copy of it. Did any of the treatment or placebo group subjects reverse their CAC score over 12 or 24 months?

3. Which if any of the items discussed in Vitamin K2 and your brain were suggested to achieve a goal of reversing existing arterial calcification?

I’ve taken a higher MK-7 daily dose (600 mcg) over the past few years. I take it with a fat source, and without vitamins A, D3, and E to eliminate the possibility of competition from other fat-soluble vitamins.

I haven’t ever had CAC measured because I’m not particularly concerned about existing arterial calcification. I’ll update this post if needed after one of the trial’s researchers provides a copy of their paper.

Eat broccoli sprouts to alleviate diabetic heart disease

gettingwell4 4-5 stars Advanced knowledge, Epigenetics, Stress ,

A 2025 rodent study investigated sulforaphane’s effects on diabetic cardiomyopathy:

“The protective effect of cruciferae-derived sulforaphane (SFN) on diabetic cardiomyopathy (DCM) has garnered increasing attention. However, no studies have specifically explored its mechanistic involvement in cardiac substrate metabolism and mitochondrial function.

To address this gap, Type 2 diabetes mellitus (T2DM) db/db mice were orally gavaged with vehicle or 10 mg/kg body weight SFN every other day for 16 weeks, with vehicle-treated wild-type mice as controls. SFN intervention (SFN-I) alleviated hyperglycemia, dyslipidemia, HOMA-IR, serum MDA levels, and liver inflammation. SFN-I improved the lipotoxicity-related phenotype of T2DM cardiomyopathy, manifested as attenuation of diastolic dysfunction, cardiac injury, fibrosis, lipid accumulation and peroxidation, ROS generation, and decreased mitochondrial complex I and II activities and ATP content.

Although not fully understood, multiple systemic and cardiac-local mechanisms contribute to DCM, encompassing hyperglycemia, dyslipidemia, insulin resistance (IR), disturbances in cardiac substrate metabolism, lipotoxicity, glucotoxicity, mitochondrial dysfunction, oxidative stress, inflammation, and endoplasmic reticulum (ER) stress. Nrf2 and its downstream metallothionein also mediated the preventive effect of SFN on DCM, and may underlie the synergistic effect of SFN and zinc in DCM.

These results suggest that chronic oral SFN-I protects against DCM by mitigating overall metabolic dysregulation and inhibiting cardiolipotoxicity. The latter might involve controlling cardiac fatty acid metabolism and improving mitochondrial function, rather than promoting glucose metabolism.”

https://www.mdpi.com/2076-3921/14/5/603 “Oral Sulforaphane Intervention Protects Against Diabetic Cardiomyopathy in db/db Mice: Focus on Cardiac Lipotoxicity and Substrate Metabolism”

This study had numerous charts like the above showing it was better to not have a deviation from health (Ctrl) rather than incur injury (DCM) then try to fix it with sulforaphane (DCM + SFN). But the control group was wild-type mice, not mice genetically inclined to diabetes like the treatment groups.

The subjects’ starting points were at nine-weeks-old (equivalent to 18-25 year-old humans), and duration was 16 weeks. Grok 3 said: “A 25-week-old db/db mouse is roughly equivalent to a human aged 30–35 years chronologically, though its metabolic condition may mimic older human physiological states in diabetes and obesity research.”

A human equivalent of a 10 mg/kg sulforaphane dose is (.081 x 10 mg) = 56 mg orally administered every other day. That’s about how much total sulforaphane I estimated I took every day (52 mg) from Week 6 through Week 56 by eating microwaved broccoli sprouts twice daily.

No rationale was provided for the sulforaphane dose or the every-other-day dosing regimen. Since I took ~52 mg every day for almost a year, I’ll guess that this study may have had more definitive results with daily dosing. Or maybe add zinc per Zinc and broccoli sprouts – a winning combination.

Nrf2 activators and transcriptomic clocks

gettingwell4 2-3 stars Required further work, Epigenetics, Stress , , , , ,

Two preprint studies looked at making transcriptional aging clocks using Nrf2 activators. Let’s start with a 2025 nematode study that used constant exposure to sulforaphane at different concentrations:

“To explore the potential of sulforaphane as a candidate natural compound for promoting longevity more generally, we tested the dose and age-specific effects of sulforaphane on C. elegans longevity, finding that it can extend lifespan by more than 50% at the most efficacious doses, but that treatment must be initiated early in life to be effective. We then created a novel, gene-specific, transcriptional aging clock, which demonstrated that sulforaphane-treated individuals exhibited a “transcriptional age” that was approximately four days younger than age-matched controls, representing a nearly 20% reduction in biological age.

The clearest transcriptional responses were detoxification pathways, which, together with the shape of the dose-response curve, indicates a likely hormetic response to sulforaphane. The hormetic, stress-pathway inducing properties of sulforaphane may indicate that many beneficial dietary supplements work in a fairly generic fashion as mild toxins rather than being driven by the biochemical properties of the compounds themselves (e.g., as antioxidants).

These results support the idea that robust longevity-extending interventions can act via global effects across the organism, as revealed by systems level changes in gene expression.”

https://www.biorxiv.org/content/10.1101/2025.05.11.653363v1 “The broccoli derivative sulforaphane extends lifespan by slowing the transcriptional aging clock”

There are difficulties in researchers translating nematode studies to mammals and humans. Nematodes lack a homolog to the Keap1 protein, which is sulforaphane’s main mammalian target to activate Nrf2.

A 2024 study developed various mammalian epigenetic clocks:

“A unified transcriptomic model of mortality that encompasses both aging and various models of lifespan-shortening and longevity interventions (i.e., mortality clocks) has been lacking. We conducted an RNA-seq analysis of mice subjected to 20 compound treatments in the Interventions Testing Program (ITP).

We sequenced the transcriptomes of a large cohort of ITP mice subjected to various neutral and longevity interventions, expanded the dataset with publicly available gene expression data representing organs of mice and rats across various strains and lifespan-regulating interventions, connected these models with survival data, and performed a meta-analysis of aggregated 4,539 rodent samples, which allowed us to identify multi-tissue transcriptomic signatures of aging, mortality rate, and maximum lifespan.

Aging and mortality were characterized by upregulation of genes involved in inflammation, complement cascade, apoptosis, and p53 pathway, while oxidative phosphorylation, fatty acid metabolism, and mitochondrial translation were negatively associated with mortality, both before and after adjustment for age.

Utilizing the aggregated dataset, we developed rodent multi-tissue transcriptomic clocks of chronological age, lifespan-adjusted age, and mortality. While the chronological clock could distinguish the effect of detrimental genetic and dietary models, it did not show a decrease in biological age in response to longevity interventions. In contrast, clocks of lifespan-adjusted age and mortality both captured aging-associated dynamics and correctly predicted the effect of lifespan-shortening and extending interventions.

Transcriptomic biomarkers developed in this study provide an opportunity to identify interventions promoting or counteracting molecular mechanisms of mortality, and characterize specific targets associated with their effects at the level of cell types, intracellular functional components, and individual genes. Our study underscores the complexity of aging and mortality mechanisms, the interplay between various processes involved, and the clear potential for developing therapies to extend healthspan and lifespan.”

https://www.biorxiv.org/content/10.1101/2024.07.04.601982v1.full “Transcriptomic Hallmarks of Mortality Reveal Universal and Specific Mechanisms of Aging, Chronic Disease, and Rejuvenation”

This second study’s references included an ITP study curated in Astaxanthin and aging, which stated:

“Despite the fact that the average diet contained 1840 ppm astaxanthin (only 46% of the target), median lifespans of male UM-HET3 mice were significantly improved. Amounts of dimethyl fumarate (DMF) in the diet averaged 35% of the target dose, which may explain the absence of lifespan effects.”

So screw-ups in making both astaxanthin and DMF mouse chows ended up with study data that didn’t measure the full lifespan impacts of activating transcription factor Nrf2. I’ll assert that such faulty data may have deviated this second study by downplaying Nrf2 activation’s impact on aging, chronic disease, and rejuvenation.

Sponsors may be less likely to be presented sulforaphane and other Nrf2 activator candidates for future aging and chronic disease studies as this first study suggests, thinking that these have already been studied in mammals. Well, maybe these compounds haven’t been accurately studied. There’s no effective way to fix a rodent study’s missing DMF Nrf2 data and faulty astaxanthin Nrf2 data to train an epigenetic clock in this second study.

I could be wrong about this second study using faulty astaxanthin Nrf2 data. It was cited as Reference 27 in the Introduction as an ITP study, but not specifically cited in the Method section. I don’t know how findings such as one of Nrf2’s target genes (“Remarkably, one of the top genes positively associated with maximum lifespan and negatively associated with chronological age and expected mortality was Gpx1, encoding the selenoprotein glutathione peroxidase 1″) and a Nrf2 specific pathway (Phase II) (“Pathways positively associated with lifespan and negatively with mortality, both before and after adjustment for age, included..xenobiotic metabolism..”) were made without Reference 27. Neither of the above studies has been peer reviewed yet.

The third phase of reversing aging and immunosenescent trends

gettingwell4 5+ stars Premium, Brain development, Epigenetics, Immune system , , , ,

Here’s a 2025 interview with Dr. Greg Fahy:

“We found that we could statistically demonstrate thymic regeneration morphologically on single individuals at single time points. MRI changes really are detecting shifts from the fatty tissue infiltration state of the involuted thymus to the regenerated thymus with functional thymic epithelial cells.

When you go through puberty your thymus involutes so you don’t have much left even when you’re 40. Essentially the process consists of loss of functional thymic mass and replacement of that functional thymic mass with adipose tissue, that’s what thymic involution is. It continues throughout life, but you retain a small amount of functional thymic mass all the way out to the age of 107.

The function of the thymus is to essentially manufacture half of your immune system. You have precursor cells arise from the bone marrow. They either go into the meiotic lineage and turn into the innate immune system, or you have the lymphocytic cells for what turns into T cells that enter the thymus and are educated in the thymus to grow up into newborn T cells and they’re released into the bloodstream.

The thymus has two jobs. It manufactures these lovely T cells without which you die but it also has a secondary finishing school. In the thymus cortex you manufacture all these lovely T cells but in the thymus medulla the T cells go to the medulla and if they don’t pass the second examination that they have to pass before they release into the body they’re all killed off. That second examination is: Do you reject self? As we get older, the thymus weakens in both the functions of making the T cells and screening out the ones that attack self. It stands to reason as we get older and the thymus’ influence wanes, we’re going to get more autoimmune disorders.

It took people a while to catch on to the fact that this involution problem is really a significant issue because the T cells that you made when you were 12, and even 20 and 40, they’re probably lasting until you’re 60. But at some point they don’t get replaced as fast as they’re going out of existence, and then your immune system goes off the cliff. Between the ages of 62 and 78 you lose 98% of your ability to recognize foreign antigens, and you still have a lot of capacity left.

We had nine guys in the first trial. Second trial we had 18 men 6 women and 2 controls that happen to be contemporaneous with that group. We have some more controls now that are either finished or or nearing completion. The second population was older than the first population by about nine years, but based on the epigenetic clocks that we looked at, they were starting off biologically younger.

On this last data analysis for Triim XA we looked at 21 different aging clocks. One aspect of the noise that we’re talking about is that biological aging as measured by some of these clocks is circadian. If you measure your age at 4:00 a.m. versus 11:00 a.m. you’re going to get a different result. It’s dynamic and there’s a trend and over time you change in a certain direction, but over any short period of time you can bounce around a little bit. The clocks predict your probability of cognitive dysfunction, they predict your probability of having impairments in your daily life, and they also predict your mortality.

We’re pretty much wrapping up that second clinical trial and going into the third. As we look at more data we understand more and more things and we see more and more things that we previously were not aware of. We began to look at a phenomena that may be responsible for limiting the magnitude of responses that we’re seeing limiting the aging reversal.

Triim-XD which is the next flavor of Triim-X is going to be looking at shifting biochemical pathways in such a way that it optimizes effects of these three medications that we’re giving people [human growth hormone, DHEA, and metformin] and prevents contradictions between them and prevents side effects of each one of these things. That’s about all I can tell you right now.”

Charts regarding the discussed item of how long effects may last are covered in The next phase of reversing aging and immunosenescent trends which was the last time I curated this research effort.

Practice what you preach, or shut up

gettingwell4 0-1 star Wasted resources, Acetylcholine, Amygdala, Anterior cingulate cortex, Beliefs, Brain development, Brain hemispheres, Brainstem, Cerebellum, Cerebrum, Cortisol, Dopamine, Epigenetics, Glutamate, Hippocampus, Hypothalamus, Immune system, Limbic system, Locus coeruleus, Lower brain, Memory, Neurochemicals, Serotonin, Stress, Thalamus , , , , , ,

A 2025 review subject was sulforaphane and brain health. This paper was the latest in a sequence where the retired lead author self-aggrandized his career by citing previous research.

He apparently doesn’t personally do what these research findings suggest people do. The lead author is a few weeks older than I am, and has completely white hair per an interview (Week 34 comments). I’ve had dark hair growing in (last week a barber said my dark hair was 90%) since Week 8 of eating broccoli sprouts every day, which is a side effect of ameliorating system-wide inflammation and oxidative stress.

If the lead author followed up with what his research investigated, he’d have dark hair, too. Unpigmented white hair and colored hair are both results of epigenetics.

Contrast this lack of personal follow-through of research findings with Dr. Goodenowe’s protocol where he compared extremely detailed personal brain measurements at 17 months and again at 31 months. He believes enough in his research findings to personally act on them, and demonstrate to others how personal agency can enhance a person’s life.

It’s every human’s choice whether or not we take responsibility for our own one precious life. I’ve read and curated on this blog many of this paper’s references. Five years ago for example:

So do more with their information than just read.

https://www.mdpi.com/2072-6643/17/8/1353 “Sulforaphane and Brain Health: From Pathways of Action to Effects on Specific Disorders”

Year Five of Changing to a youthful phenotype with sprouts

gettingwell4 4-5 stars Advanced knowledge, Epigenetics, Immune system, Memory, Stress

1. I’ve continued daily practices from Year Four to experience another year without being sick! I’ll get a set of Labcorp tests in a week to see if anything is sneaking up on me.

Really think that Brassica clinical trials should last years, not weeks. Once people get over the fact that broccoli, red cabbage, and mustard sprouts will never taste good because their compounds are plants’ defenses against predators, they’ll overlook that in favor of health benefits. Avena sativa oat sprouts don’t have a palatability problem.

2. Daily supplements have changed a little:

  • Started taking a quercetin supplement suggested in a comment to Year One as helpful for seasonal allergies (it doesn’t do that for me). Repeatedly rinsing and soaking the salt out of capers for quercetin content became too much of a nuisance, and the results didn’t always taste right;
  • Stopped taking Prodrome supplements because of unsustainable high costs;
  • Started taking Ovega 3 algae oil DHA 420 mg/EPA 140 mg twice a day in their place;
  • Substituted flax oil 1400 mg once a day for Balance oil;
  • Started taking 2 g magnesium L-threonate;
  • Upped taurine intake from 5 to 6 grams;
  • Upped D3 by 25 mcg to a daily 4400 IU;
  • Reduced chondroitin sulfate by 1.8 g since my joints are doing fine;
  • Stopped soy lecithin in favor of eating three raw eggs.

3. I injured my left shoulder in May 2024 by overdoing upper body exercises, and stopped seven months to recover. Gained thirty pounds during that layoff, and have worked off ten pounds with new routines since then.

I’m no longer dogmatic about aerobic exercise / beach walks. I’ll go over to the beach before sunrise when it isn’t raining or windy, or wait until the afternoon for weather to improve, rather than walk 30 minutes a day irregardless.