Nrf2 and senescence, Part 2

September 7, 2023September 7, 2023 gettingwell4 4-5 stars Advanced knowledge, Epigenetics, Immune system, Stress Control group, Genetic factors

A 2023 rodent study investigated Nrf2’s capacity to reverse cell senescence:

“Poly-D,L-lactic acid (PDLLA) filler corrects soft tissue volume loss by increasing collagen synthesis in the dermis. Adipose-derived stem cells (ASCs) are known to attenuate the decrease in fibroblast collagen synthesis that occurs during aging.

Conclusions:

PDLLA increased macrophage NRF2 expression, resulting in increased M2 polarization and IL-10 expression in senescent macrophages and aged skin.

Increased IL-10 expression by macrophages led to reduced ASC senescence, and increased ASC proliferation and paracrine secretion of TGF-β and FGF2.

Increased TGF-β and FGF2 levels in turn led to increased fibroblast proliferation and synthesis of collagen and elastin fibers.

PDLLA-modulated macrophage not only directly stimulated fibroblast activity, promoting proliferation and collagen synthesis, but also reduced expression of NF-kB and MMP2/3/9.

These effects ultimately led to skin rejuvenation in aged skin.”

https://www.mdpi.com/2076-3921/12/6/1204 “Poly-D,L-Lactic Acid Filler Increases Extracellular Matrix by Modulating Macrophages and Adipose-Derived Stem Cells in Aged Animal Skin”

This study’s Nrf2 activator was persistent. Per Part 1’s findings, that probably won’t work out well long-term, as constant Nrf2 activation may cause cell senescence. It would still be worthwhile for other researchers to replicate this study’s rejuvenation effects using other Nrf2 activators with different activation periods and additional senescence measurements.

It was this study’s H₂O₂ and unmeasured aging environments that caused cell senescence. Other studies could follow principles of An environmental signaling paradigm of aging and rejuvenate by changing subjects’ aging environments since:

“It is clear that the increasing number of senescent cells depends on the post-adult developmental stage rather than chronological age. The coincidence that these processes result in particular forms of impairment in old age does not seem to be random as it is present in all mammals, and may be causative of many aspects of aging.”

Maybe similar to how Environmental signaling rescues aging muscle stem cells reversed aged muscle stem cell gene expression?

Nrf2 and senescence, Part 1

September 6, 2023September 11, 2023 gettingwell4 4-5 stars Advanced knowledge, Epigenetics, Immune system, Stress Control group, Genetic factors, Neurodegenerative disease

A 2023 rodent study investigated Nrf2’s capacity to cause cell senescence:

“The KEAP1-NRF2 pathway is a stress response pathway which has been maintained by natural selection due to its ability to benefit survival of the host organism. One important distinction between this pathway and other stress response pathways such as p53, is that chronic activation of NRF2 has not been associated directly with a mechanism to promote cell death if survival of the cell becomes deleterious to the host.

Some unexplained observations suggest that NRF2 activation has additional physiological outputs which have yet to be described. For example, despite the fact that oxidative stress plays an important role in etiology of many aging-related diseases, genetic activation of Nrf2 in mice is associated with decreased lifespan.

We found that NRF2 functions to prime cells to become senescent in response to irreparable damage. In diseased states, NRF2 promotes transcriptional activation of a specific subset of the senescence-associated secretory phenotype (SASP) gene program, which we have named the NRF2-induced secretory phenotype (NISP).

As Nrf2 also promotes monocyte and macrophage invasion in mouse disease models of steatohepatitis, colitis, pancreatitis, and autoimmune nephritis, we would posit that it represents a central component of the Nrf2 response in damaged epithelial tissues, and that the NRF2-NISP-Immune recruitment model represents a framework through which these disease phenotypes can be understood.

This pathway represents the final stage of the oxidative stress response, as it allows cells to be safely removed if macromolecular damage caused by the original stressor is so extensive that it is beyond the repair capacity of the cell.”

https://www.sciencedirect.com/science/article/pii/S221323172300246X “A NRF2-induced secretory phenotype activates immune surveillance to remove irreparably damaged cells”

Continued in Part 2.

Ripe wild grapes

Fructose and survival

September 4, 2023September 4, 2023 gettingwell4 4-5 stars Advanced knowledge, Cerebrum, Epigenetics, Hippocampus, Hypothalamus, Immune system, Limbic system, Memory, Neurochemicals, Stress, Vasopressin Genetic factors, Neurodegenerative disease

This 2023 paper provided mechanistic evidence, evolutionary theory, and testable scenarios for fructose metabolism differences from other nutrients:

“The fructose survival hypothesis proposes that obesity and metabolic disorders may have developed from over-stimulation of an evolutionary-based biologic response (survival switch) that aims to protect animals in advance of crisis. The response is characterized by hunger, thirst, foraging, weight gain, fat accumulation, insulin resistance, systemic inflammation, and increased blood pressure.

Unlike other nutrients, fructose reduces the active energy (adenosine triphosphate) in the cell, while blocking its regeneration from fat stores. This is mediated by intracellular uric acid, mitochondrial oxidative stress, inhibition of AMP kinase, and stimulation of vasopressin.

Fructose metabolism is associated with oxidative stress, mitochondrial dysfunction, loss of cytoprotective transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2), and a reduction in sirtuins that characterize the ageing process. Fructose also induces generation of advanced glycation end products much more effectively than glucose.

The fructose pathway is almost inevitably strongest in early disease states, for over time there is often fibrosis, inflammation, or mitochondrial loss that results in persistence of the disease process. The best time for intervention may turn out to be in early disease before conditions become less reversible.”

https://royalsocietypublishing.org/doi/10.1098/rstb.2022.0230 “The fructose survival hypothesis for obesity”

Time to exit fructose survival mode.

Part 2 of Harnessing endogenous defenses with broccoli sprouts

August 31, 2023 gettingwell4 5+ stars Premium, Epigenetics, Immune system, Stress Genetic factors

The author of this 2023 paper expanded Part 1 to include further clinical evidence and four human case studies. I’ll highlight just a few items because it’s quite detailed:

“Accumulating evidence for the crucifer-derived bioactive molecule sulforaphane (SFN) in upstream cellular defence mechanisms highlights its potential as a therapeutic candidate in targeting functional gastrointestinal conditions, as well as systemic disorders. This article catalogues evolution of and rationale for a hypothesis that multifunctional sulforaphane can be utilised as the initial step in restoring ecology of the gut ecosystem.

It can do this primarily by targeting functions of intestinal epithelial cells. In many cases where primary presenting symptoms are related to aberrant intestinal function, complete or partial resolution also occurred in seemingly unrelated conditions such as inflammatory skin diseases, multiple food intolerances, histamine-like allergic reactions, and neuro-psychological disorders.

Although SFN was the primary and initial intervention, clinicians recommended that their patients consume a mixed diet of minimally processed foods rich in vegetables and other sources of phytochemicals. It was also clear that dietary recommendations alone were not capable of making changes that occurred when SFN was added.

In seeking an effective gateway for addressing digestive, immune, cardiometabolic and other chronic disease, this hypothesis proposes an approach that harnesses the endogenous processes of human cells. These processes focus on restoring homeostasis to the gut, its underlying immune network, and the companion microbiota, with the collective potential to beneficially impact all gut–organ axes.”

https://www.mdpi.com/1422-0067/24/17/13448 “The Rationale for Sulforaphane Favourably Influencing Gut Homeostasis and Gut–Organ Dysfunction: A Clinician’s Hypothesis”

The author proposed this paper as a working hypothesis to be scientifically correct. Would a null hypothesis be along the lines of “I’ll eat a clinically relevant dose of broccoli sprouts every day for twelve weeks, and nothing will change”?

Case study #1 had a timing parallel with my experiences per:

“She was able to tolerate 20 mg SFN daily after several weeks; the dose was increased to 40 mg daily by 6 weeks.”

I doubled my dose at Week 6. All case studies documented transformative experiences, but they weren’t the same types that shortly followed for me.

Acetyl-L-carnitine dosing

August 30, 2023 gettingwell4 4-5 stars Advanced knowledge, Beliefs, Cerebrum, Epigenetics, Stress Brain neurons, Control group, Genetic factors, Levels of consciousness, Neurodegenerative disease

Haven’t curated acetyl-L-carnitine papers recently. Here are three 2023 studies, beginning with a human case report:

“It is believed that 75% of the required amount of carnitine is taken from diet and the remaining 25% is synthesized in the body. Long-term use of a carnitine-free diet is thought to increase the risk of carnitine deficiency.

Dosage for long-term tube-fed patients with disorders of consciousness and convulsive seizures, such as in the present cases, is not specified. Instructions accompanying the medication list gastrointestinal symptoms such as nausea, vomiting, and diarrhea as side effects of L-carnitine. They indicate a maximum dosage of 3 g/day, and a maximum single dose of 1 g.

L-Carnitine is efficiently absorbed in the gastrointestinal tract when taken in small amounts, but when taken in large amounts, the transporter is saturated and bioavailability is only about 10%–20%. Although safety of oral L-carnitine administration is considered high because there is an upper limit to the amount that can be absorbed, clinicians should remain aware of side effects noted above.

To the best of our knowledge, this is the first report in which L-carnitine was administered to a patient with impaired consciousness after stroke with the result that symptoms improved. It is possible that carnitine deficiency is overlooked in some patients in rehabilitation wards, and measurement of ammonia may be useful in its detection. Because carnitine deficiency might interfere with active rehabilitation, nutritional management with attention to carnitine deficiency is important in rehabilitation wards.”

https://www.jstage.jst.go.jp/article/prm/8/0/8_20230019/_html/-char/en “Disorders of Consciousness after Subacute Stroke Might Partly be Caused by Carnitine Deficiency: Two Case Reports”

I currently take one gram of acetyl-L-carnitine three times a day.

Next is a clinical trial with amyotrophic lateral sclerosis (ALS) patients that used two different doses of acetyl-L-carnitine:

“Our findings did not confirm an effect of ALCAR 3 g/day on survival in ALS subjects at 24 months. An effect was observed in those treated with ALCAR 1.5 g/day.

In addition, we did not detect an effect on self-sufficiency at 12 months as previously seen in the pilot trial. These differences could be explained by:

The study design (retrospective observational study vs prospective randomized trial);

Selection bias (subjects from the real-world clinical practice are less selected than those included in a clinical trial); and

Drug compliance (subjects enrolled in a clinical trial perform several onsite evaluations in which compliance is verified by tablets accounting, while in clinical practice this is not done).

Our hypothesis is that the presence of residual confounding might explain our unexpected results. Residual confounding refers to the presence of an unmeasured or uncontrolled variable that could affect the relationship between treatment (ALCAR) and outcome.

This study provided additional information on the potential effect of ALCAR on disease progression and survival, and adds evidence to justify the use of ALCAR in ALS subjects.”

https://link.springer.com/article/10.1007/s00415-023-11844-6 “Retrospective observational study on the use of acetyl-L-carnitine in ALS”

This study’s dosing method wasn’t clear on exactly how doses were administered every day. I’ll guess that if both 1.5 and 3 grams were given all at once, they might have been roughly equivalent doses per the first paper’s cited bioavailability saturation effect.

Next is a rodent aging study:

“The aim of this study was to examine effects of long-term L-Carnitine (β-hydroxy-γ-trimethylaminobutyric acid, LC) administration on cardiomyocyte contraction and intracellular Ca²⁺ transients in aging rats. LC (50 mg/kg body weight/day) was dissolved in distilled water and orally administered for a period of 7 months.

LC increased cardiomyocyte cell shortening and resting sarcomere length. LC supplementation led to a reduction in resting [Ca²⁺]i level and an increase in the amplitude of [Ca²⁺]i transients, indicative of enhanced contraction. Consistent with these results, decay time of Ca²⁺ transients also decreased significantly in the LC-treated group.

Long-term administration of LC may help restore Ca²⁺ homeostasis altered during aging, and could be used as a cardioprotective medication in cases where myocyte contractility is diminished.”

https://link.springer.com/article/10.1007/s00418-023-02215-3 “L-Carnitine improves mechanical responses of cardiomyocytes and restores Ca²⁺ homeostasis during aging” (not freely available)

A human equivalent of this study’s daily dose is (50 mg x .162) x 70 kg = 567 mg. A human equivalent of this study’s duration using the maximum lifespan method is (7 months x 32.2) = 225.4 months. The subjects began at 11 months old (human equivalent age 29.5 years) and ended at 18 months old (human equivalent age 48.3 years).

This study illustrated how heart dysfunctions with subclinical symptoms advance with aging, and that starting to do something preventative before human equivalent age 30 may work.

Fermented oats

August 25, 2023August 25, 2023 gettingwell4 4-5 stars Advanced knowledge, Epigenetics, Immune system, Stress Control group, Genetic factors

This 2023 review subject was fermented oats as food:

“We provide a comprehensive overview of fermented oat products available on the market, and various production methods employed for fermenting oats. We investigate how fermentation affects the chemical composition and biological functions of oats.

Increased nutritional content of fermented oats is associated with various health benefits, including anti-inflammatory and antioxidant activities. Further investigations are warranted to elucidate nutritional benefits of fermented oats in human nutrition.”

https://www.mdpi.com/2072-6643/15/16/3521 “Fermented Oats as a Novel Functional Food”

I’ve had trouble sprouting Avena sativa oats received earlier this month via Amazon. They’ve gone from >90% sprouting over three days to <10%.

These batches’ fermenting mixtures after three days have been on their way to becoming mash for brewing. They’ve tasted sour, but not objectionable to where I’ve thrown away a batch. This review showed how fermented oats may have value in certain areas.

I didn’t have this happen with Avena sativa oats ordered in April 2023, December 2022, and July 2022. I did have this happen before with a September 2021 order. After I complained to the Montana farmer, they sent me a sample of what they shipped to Amazon, and those oats sprouted as expected.

It seems that Amazon’s iffy pallet handling and storage during hot weather doesn’t always preserve the seed vitality needed for sprouting. l’ll place a larger order come December this year, as I’d rather have sprouted oats’ benefits.

How long does sulforaphane keep?

August 22, 2023September 19, 2023 gettingwell4 2-3 stars Required further work Neurodegenerative disease

This 2023 clinical trial with broccoli sprout powder investigated ways of improving blood plasma measurements:

“Quantifying sulforaphane (SFN) and its thiol metabolites in biological samples using liquid chromatography-tandem mass spectrometry is complicated by SFN’s electrophilic nature and the facile dissociation of SFN-thiol conjugates. We used the alkylating agent iodoacetamide (IAA) to both release SFN from protein thiols and force the dissociation of SFN metabolites.

After 15 min of incubation, approximately 90% of the SFN was recovered. After 2 hours of incubation, SFN loss to thiol conjugation was significant, with approximately 30% recovered. The percentage of SFN lost in this manner in human plasma upon protein precipitation would likely be greater than the loss we observed in fetal bovine serum (FBS), as free thiols in human plasma are approximately 6 times higher than in FBS.

Subjects were 12 healthy, young (19–30 y), men (n = 4), and women (n = 8). They consumed three EnduraCell capsules with water.

Per the manufacturer, each capsule contained 700 mg of 100% whole broccoli sprout powder, including active myrosinase and 21 mg of glucoraphanin, which upon full conversion to SFN would yield ∼8 mg, equaling ∼24 mg of SFN total per three-capsule dose. We note that full conversion to SFN, even with active myrosinase in the supplement, is not expected.

SFN concentrations in plasma increased for all subjects, with the highest being 193 nM for subject 4 one hour post consumption. Asterisks for the 2 and 3 hour time points indicate a p-value of <0.05, comparing that time point to the previous time point for that participant.

This thiol-blocking method increased SFN percent recovery from FBS from 32 to 94 ± 5%. Applying the method to clinical samples, SFN concentrations were on average 6 times greater than when IAA was omitted.”

https://pubs.acs.org/doi/10.1021/acs.jafc.3c01367 “Bioavailable Sulforaphane Quantitation in Plasma by LC–MS/MS Is Enhanced by Blocking Thiols”

I didn’t highlight subject bioavailability statistics or researcher generalizations. Someday, researchers will be interested and forward-thinking enough about their field to plan ahead and investigate likely occurrences such as:

What caused Subjects 3 and 4 to have much higher responses than Subjects 2 and 6?
What caused Subjects 5 and 11 to have their highest sulforaphane levels at their 3-hour points, whereas 3-hour points were most other subjects’ lowest levels?

And, of course, what was the actual sulforaphane intake by researcher measurements vs. relying on manufacturer statements? Especially when researcher lab equipment was available to measure that.

This study’s proof of concept demonstrated a 10% sulforaphane loss after 15 minutes, and a 68% loss after two hours. Applying their finding to home sprouting, I wouldn’t expect any advance preparation method of broccoli sprouts or seeds to adequately address sulforaphane degradation.

I’d suggest microwaving up to 60°C (140°F) and/or grinding and/or blending broccoli sprouts and seeds just before eating to optimize your sulforaphane intake. Also, taking the very reactive sulforaphane by itself to optimize your results.

Take Vitamin K2 to protect against aluminum toxicity

August 16, 2023August 17, 2023 gettingwell4 4-5 stars Advanced knowledge, Acetylcholine, Cerebrum, Dopamine, Epigenetics, Hippocampus, Limbic system, Memory, Neurochemicals, Stress Brain neurons, Control group, Neurodegenerative disease, Prefrontal cortex

This 2023 rodent study investigated relationships of MK-7 menaquinone, aluminum trichloride, and brain health:

“A variety of endogenous and exogenous agents, such as metals and environmental toxins (aluminum, mercury, etc.), can contribute to neurodegeneration, which is of multifactorial clinical occurrence.

The current study showed that Alzheimer’s Disease (AD)-like condition was induced in mice by AlCl₃ treatment affecting spatial and recognition memory. Neuropathological alterations included neuroinflammation, oxidative stress, an increase in brain amyloid β levels, and loss of hippocampal neurons.

Aluminium chloride (AlCl₃; 100 mg/kg for 3 weeks orally) was administered to Swiss albino mice to induce neurodegeneration and Vitamin K₂ (100 mcg/kg for 3 weeks orally) was applied as treatment. This was followed by behavioral studies to determine memory changes.

Antioxidants like glutathione and SOD were low compared to the control group, while oxidative stress marker MDA was elevated. BDNF levels increased in the Vitamin K₂ treated animals, suggesting its neuroprotective functions.

Vitamin K₂ could partially reverse AlCl₃-mediated cognitive decline. It increased hippocampal acetylcholine and BDNF levels while reducing oxidative stress, neuroinflammation, and β-amyloid deposition, protecting hippocampal neurons from AlCl₃-mediated damage.“

https://link.springer.com/article/10.1007/s10787-023-01290-1 “Vitamin K2 protects against aluminium chloride-mediated neurodegeneration” (not freely available)

This study’s human equivalent Vitamin K₂ dose is (100 mcg x .081) x 70 kg = 567 mcg. I’ve taken 600 mcg MK-7 every day for the past two years.

Found out last week that I’ve also been inadvertently dosing myself with aluminum every day. This is the underside of my former 3-year-old drip coffee maker with its cover removed:

I’m certain its aluminum tubing that heats reservoir water started to corrode a long time ago. Currently trying out methods of making aluminum-free coffee.

Reversing biological age in rats

August 12, 2023August 12, 2023 gettingwell4 4-5 stars Advanced knowledge, Brain development, Cerebellum, Cerebrum, Epigenetics, Hippocampus, Hypothalamus, Limbic system, Lower brain, Memory, Stress, Telomeres Brain neurons, Control group, DNA methylation, Genetic factors, Happiness, Neural plasticity, Neurodegenerative disease, Prefrontal cortex, Reputations

This 2023 rodent study wrapped together findings of the original study curated in A rejuvenation therapy and sulforaphane, and the second follow-on study mentioned in Signaling pathways and aging. I’ll start by highlighting specifics of the later study:

“Pronounced rejuvenation effects in male rats prompted us to conduct further confirmatory experiments. A particularly important consideration is the effectiveness of E5 with regards to sex, as sex-dependent rejuvenation by some interventions have previously been reported.

To assess E5’s applicability to both male and female Sprague Dawley rats, we studied 12 males (6 treated with E5, 6 with saline) and 12 females (6 treated with E5, 6 with saline). These rats were treated every 45 days with an injection of E5 or saline. Rats were monitored for 165 days, and blood was drawn at six time points: 0, 15, 30, 60, 150 and 165 days from the first injection.

We observed highly significant improvements in TNF alpha and IL-6 levels for both males and females in the blood of E5-injected rats over that of saline controls. We also observed a substantial improvement in grip strength.

Our study shows age reversal effects in both male and female rats, but E5 is more effective in males.”

Another experimental group was started with old rats of both sexes. Using the human / rat relative clock developed in the original study, a human equivalent age to these rats at 26 months old was ((112.7 weeks / 197.6 weeks maximum rat lifespan) x 122.5 years maximum human lifespan) = 69.8 years:

“To validate our epigenetic clock results, we conducted a second set of E5 experiments with Sprague Dawley rats of both sexes. When these rats turned 26 months old, half (9 rats) received the E5 treatment while the other half (8 rats) received only the control treatment (saline injection). We analyzed methylation data from two blood draws: blood draw before treatment (baseline) and a follow up sample (15 days after the E5/saline treatment).”

Treatment measurements were affected by one female control group outlier. Panels F through J were recalculated after removing the outlier to show significant effects in both sexes:

“A) Final version of the rat clock for blood. Baseline measurement (x-axis) versus follow up measurement (15 days after treatment, y-axis). Points (rats) are colored by treatment: red=treated by E5, black=treated with saline only. Rotated grey numbers underneath each bar reports the group sizes. Each bar plot reports the mean value and one standard error.

B,D,E) Difference between follow up measurement and baseline measurement (y-axis) versus treatment status in B) all rats, D) female rats only, E) male rats only. C) is analogous to B) but uses the pan tissue clock for rats.

Panels in the second row (F,G,H,I,J) are analogous to those in the first row but the analysis omitted one control rat (corresponding to the black dot in the lower right of panel A).”

https://www.biorxiv.org/content/10.1101/2023.08.06.552148v1 “Reversal of Biological Age in Multiple Rat Organs by Young Porcine Plasma Fraction”

A description of how E5 plasma fraction was made starts on page 16 of the *.pdf file. The next E5 study will be done with dogs per July 2023 updates in blog post comments:

“On E5 our entire team is working hard towards the launch of an old Beagle dogs trial this month. We want to make them really young, healthy, happy, and jumping around like 1 and 2 year olds.

Primary endpoint is safety and toxicology to test various dose strengths and frequencies. Secondary endpoints are more than 20.

As you know, we like to test exhaustively to get a sharper perspective of what’s happening. In rat studies we tested 30 biomarkers, including functional. We are especially keen to check kidney markers.

There are two clocks for dogs we are interested in to get third party confirmation of age reversal. Horvath dog clock is ready and GlycanAge dog clock is under construction.

We are requesting all organizations that support pets and aging to financially support their project of building an accurate dog clock. Not only will it help veterinary aging research like ours, but also all the dog owners that may want to know how much improvement their dog received from treatment. Dr. Matt Kaeberlain is an advisor on their project.”

36 holes in your roof

August 9, 2023August 11, 2023 gettingwell4 4-5 stars Advanced knowledge, Cerebrum, Cortisol, Hippocampus, Immune system, Limbic system, Memory, Neurochemicals, Stress, Telomeres, Testosterone Brain neurons, Control group, DNA methylation, Genetic factors, Inherited disease, Neurodegenerative disease

An August 2023 interview with Dr. Dale Bredesen, who has reversed Alheizmer’s disease in many people, which will never be acknowledged by the corrupt paradigm:

“How much do you want me to go into things that are relatively controversial and how much do you want me to stick with kind of the more standard line?

For Alzheimer’s we noticed initially there are 36 different potential contributors. You need to patch as many as possible to have an effect.

All of these things, your estradiol level, your progesterone level, pregnenolone, free T3, TSH, Vitamin D, testosterone, these things are all critical. They all feed into the equation.

You have over a hundred trillion contacts in your brain. Will you be able to keep them? Or do you not have what it takes to keep them, and you have to downsize?

The reality is Alzheimer’s disease should be a rare disease. If everybody would get on appropriate prevention or early reversal, we could make it a rare disease.”

https://brokenscience.org/podcasts-ep-5/ “Dale Bredesen – Reversing Alzheimer’s Fate”

See A therapy to reverse cognitive decline for previous curation of Dr. Bredesen’s work.

Eat broccoli sprouts to prevent thrombosis

August 8, 2023 gettingwell4 4-5 stars Advanced knowledge, Stress Control group, Genetic factors

This 2023 cell / rodent study investigated dietary plant compounds for their functions related to blood clots after emergencies like heart attack and stroke:

“We evaluated phenotypes associated with irreversible protein engagement of twenty-three electrophilic phytochemicals. This revealed a novel antiplatelet selectivity profile of natural product sulforaphane (SFN).

Response of platelets to adenosine diphosphate and a thromboxane A2 receptor agonist was impaired without affecting thrombin and collagen-related peptide activation. SFN also substantially reduced formation of platelet thrombi on surfaces coated with collagen under arterial flow conditions.

SFN displayed important characteristics of prophylactic agents. It was able to improve clot-busting performance of recombinant tissue plasminogen activator (rtPA) in an in vivo electrolytic injury model of thrombosis without increasing blood loss.

All current antiplatelet agents are contraindicated for adjunctive therapies for thrombolysis in stroke patients, due to the high risk of symptomatic brain hemorrhage, the most feared complication of thrombolytic therapy. Our results serve as a catalyst for further investigations into preventive and therapeutic mechanisms of dietary antiplatelets, with a view to develop more effective and safer adjunctive treatments to improve clot-busting power of rtPA – currently the sole approved therapeutic for stroke recanalization that has significant limitations.”

https://chemrxiv.org/engage/chemrxiv/article-details/64a2ca49ba3e99daef721461 “Integrating Phenotypic and Chemoproteomic Approaches to Identify Covalent Targets of Dietary Electrophiles in Platelets”

The era of following wise old men ended a long time ago

August 3, 2023August 4, 2023 gettingwell4 < 0 Detracted from science, Beliefs, Cerebrum, Epigenetics, Immune system, Stress Control group, Genetic factors

I try to stay away from papers that waste resources or detract from science. This 2023 lab study irked me by emphasizing risks of home sprouting without also pointing out the many benefits.

These researchers, who obviously don’t home sprout, used the supplier I get organic broccoli seeds from as a punching bag. They consulted a broccoli sprouts expert to recommend bleaching seeds before sprouting.

Fine. Do these people ever eat a salad without also bleaching those ingredients? Do they risk eating at restaurants? How do they get motivated to take the risk of leaving their dwelling/dormitory?

What did our ancestors eat? Was it luck that they didn’t exterminate themselves with their food hygiene? Or have humans adapted to dealing with all types of pathogens?

The expert is a few weeks older than I am, and has completely white hair. I’ve had dark hair since Week 8 of eating broccoli sprouts every day, which reflects ameliorating system-wide inflammation and oxidative stress. Next month will be three and a half years of this daily practice.

If the expert followed what their research investigated, they’d have dark hair, too. White hair and dark hair are both epigenetic. It’s every human’s choice whether or not we take responsibility for our own one precious life.

https://www.mdpi.com/2304-8158/12/4/747 “Seed Disinfestation Practices to Control Seed-Borne Fungi and Bacteria in Home Production of Sprouts”

Neuritogenesis

July 30, 2023July 30, 2023 gettingwell4 4-5 stars Advanced knowledge, Acetylcholine, Amygdala, Brain development, Brainstem, Cerebellum, Cerebrum, Dopamine, Epigenetics, Glutamate, Hippocampus, Hypothalamus, Immune system, Limbic system, Lower brain, Memory, Neurochemicals, Stress, Thalamus Brain neurons, Control group, Embryo development, Genetic factors, Infants, Neural plasticity, Prefrontal cortex

Three 2023 papers on the initial stage of neuronal differentiation, starting with a rodent study of taurine’s effects:

“We aimed to assess the role of taurine (TAU) in axonal sprouting against cerebral ischemic injury, clarify the function of mitochondria in TAU-induced axonal sprouting, and further determine the underlying potential molecular mechanism.

We determined that TAU improved motor function recovery and restored neurogenesis in ischemic stroke. This possibly occurred via improvements in mitochondrial function.

We investigated that the Sonic hedgehog (Shh) pathway exerted an important role in these effects. Our study findings highlighted the novel viewpoint that TAU promoted axonal sprouting by improving Shh-mediated mitochondrial function in cerebral ischemic stroke.”

https://www.scielo.br/j/acb/a/nxKvGXGk9g6gRkHxybMfbYJ/?lang=en “Taurine promotes axonal sprouting via Shh-mediated mitochondrial improvement in stroke”

A rodent study investigated effects of a soy isoflavone gut microbiota metabolite:

“Perinatally-infected adolescents living with HIV-1 (pALHIV) appear uniquely vulnerable to developing substance use disorders (SUD). Medium spiny neurons (MSNs) in the nucleus accumbens core (NAcc), an integrator of cortical and thalamic input, have been implicated as a key structural locus for the pathogenesis of SUD.

Treatment with estrogenic compounds (e.g., 17β-estradiol) induces prominent alterations to neuronal and dendritic spine structure in the NAcc supporting an innovative means to remodel neuronal circuitry. The carcinogenic nature of 17β-estradiol, however, limits its translational utility.

Plant-derived polycyclic phenols, or phytoestrogens, whose chemical structure resembles 17β-estradiol may afford an alternative strategy to target estrogen receptors. The phytoestrogen S-Equol (SE), permeates the blood-brain barrier, exhibits selective affinity for estrogen receptor β (ERβ), and serves as a neuroprotective and/or neurorestorative therapeutic for HIV-1-associated neurocognitive and affective alterations.

Beginning at approximately postnatal day (PD) 28, HIV-1 transgenic (Tg) animals were treated with a daily oral dose of 0.2 mg of SE. The SE dose of 0.2 mg was selected for two primary reasons, including:

A dose-response experimental paradigm established 0.2 mg of SE as the most effective dose for mitigating neurocognitive deficits in sustained attention in the HIV-1 Tg rat; and

The dose, which yielded a daily amount of 0.25–1.0 mg/kg/SE (i.e., approximately 2.5–10 mg in a 60 kg human), is translationally relevant (i.e., well below the daily isoflavone intake of most elderly Japanese.

Daily oral treatment continued through PD 90.

HIV-1 Tg animals exhibited an initial increase in dendrite length (A) and the number of dendritic spines (B) early in development; parameters which subsequently decreased across time. In sharp contrast, dendrite length and the number of dendritic spines were stable across development in control animals.

Targeting these alterations with the selective ERβ agonist SE during the formative period induces long-term modifications to synaptodendritic structure, whereby MSNs in the NAcc in HIV-1 Tg animals treated with SE resemble control animals at PD 180.”

https://www.degruyter.com/document/doi/10.1515/nipt-2023-0008/html “Constitutive expression of HIV-1 viral proteins induces progressive synaptodendritic alterations in medium spiny neurons: implications for substance use disorders”

A rodent brain cell study investigated soy isoflavones’ effects on a different estrogen receptor:

“We evaluated effects of isoflavones using mouse primary cerebellar culture, astrocyte-enriched culture, Neuro-2A clonal cells, and co-culture with neurons and astrocytes. Soybean isoflavone-augmented estradiol mediated dendrite arborization in Purkinje cells.

These results indicate that ERα plays an essential role in isoflavone-induced neuritogenesis. However, G-protein-coupled ER (GPER1) signaling is also necessary for astrocyte proliferation and astrocyte–neuron communication, which may lead to isoflavone-induced neuritogenesis.

We highlight the novel possibility that isoflavones enhance dendritogenesis and neuritogenesis, indicating that they can be a useful supplementary compound during brain development or in the injured brain.”

https://www.mdpi.com/1422-0067/24/10/9011 “Isoflavones Mediate Dendritogenesis Mainly through Estrogen Receptor α”

A blood plasma aging clock, Part 2

July 27, 2023 gettingwell4 2-3 stars Required further work, Epigenetics, Hippocampus, Immune system, Limbic system, Stress, Testosterone Control group, Genetic factors, Neurodegenerative disease

Quite a few people recently looked at Part 1 which curated “Undulating changes in human plasma proteome across lifespan are linked to disease” in December 2019. Let’s start with a 2023 human study coauthored by Part 1’s lead researcher:

“The aim of this study is to identify a set of proteins in human plasma associated with aging by integration of data of four independent, large-scaled datasets. We identified a set of 273 plasma proteins significantly associated with aging (aging proteins, APs) across these cohorts consisting of healthy individuals and individuals with comorbidities and highlight their biological functions.

Although these presented proteins may be different compared to other presented proteomic clocks [like Part 1’s], this can be explained due to a variety of factors. Across studies there may be several technical factors, such as used anti-coagulants, and biological differences, such as different age ranges, ethnicity and corrections for BMI, which may influence the plasma proteome in the cohorts. To overcome these differences, we focused on the overlap between the different studies as they also present several of these confounding factors.

We show that individuals presenting accelerated or decelerated aging based on their plasma proteome, respectively have a more aged or younger systemic environment. These results provide novel insights in understanding the aging process and its underlying mechanisms and highlight potential modulators contributing to healthy aging.”

https://www.frontiersin.org/articles/10.3389/fragi.2023.1112109/full “Markers of aging: Unsupervised integrated analyses of the human plasma proteome”

A 2023 human study cited the above study and found:

“Our cross-sectional study of adults adherent and non-adherent to recommended lifestyle habits established strong group differences for 39 proteins primarily related to innate immunity and lipoprotein metabolism. Many of these protein differences were best explained by group contrasts in adiposity and visceral fat. The relatively small number of upregulated and downregulated proteins associated with good lifestyle habits should facilitate development of a targeted lifestyle proteomic panel that can be used in future studies to determine efficacy of various prevention and treatment strategies.”

https://www.researchsquare.com/article/rs-3097901/v1 “Adherence to Lifestyle Recommendations Linked to Innate Immunity and Lipoprotein Metabolism: A Cross-Sectional Comparison Using Untargeted Proteomics”

A 2023 human study from Google-owned Calico:

“In most cases, direction of effects between cause-specific and all-cause mortality was concordant, but all-cause mortality association was not statistically significant. Neither do we have insight into conditional causal effects of these proteins nor interaction effects between them.”

https://www.researchsquare.com/article/rs-2626017/v1 “Plasma Proteomic Determinants of Common Causes of Mortality”

“Undulating” in Part 1 described plasma proteins changing over time with peaks at ages 34, 60, and 78. Those peaks don’t provide a base for linearly extrapolating all-cause mortality.

A 2023 rodent study did a touch better with one of Part 1’s 46 proteins of a conserved aging signature that changed in the same direction with mice and humans, although it didn’t fully investigate protein expression over time.

“Interactions between CHRDL1 levels, age, and plasma lipids that might affect cardiometabolic health should be further investigated.”

https://www.mdpi.com/2073-4409/12/4/624 “Chordin-like 1, a Novel Adipokine, Markedly Promotes Adipogenesis and Lipid Accumulation”

Sulforaphane, TFEB, and ADH1

July 24, 2023January 19, 2025 gettingwell4 4-5 stars Advanced knowledge, Epigenetics, Hippocampus, Immune system, Limbic system, Memory, Stress Brain neurons, Control group, Genetic factors, Neural plasticity, Neurodegenerative disease

Looked for a recent follow-on study of the 2021 Precondition your defenses with broccoli sprouts, specifically:

“NFE2L2/NRF2 is a target gene of TFEB (transcription factor EB), a master regulator of autophagic and lysosomal functions, which we show here to be potently activated by sulforaphane.”

Some interesting papers cited it, but no studies continued its sulforaphane/TFEB line of inquiry. A 2022 review made a good point when citing this study for TFEB, but didn’t tie it in with sulforaphane:

“TFEB is translocated into the nucleus with a moderate increase of ROS through a Ca²⁺-dependent, but mTOR (mechanistic target of rapamycin kinase)-independent mechanism. Essential genes involved in lysosome biogenesis and autophagosome are activated, which are crucial for removal of damaged mitochondria.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9730074/ “Phytochemicals and modulation of exercise-induced oxidative stress: a novel overview of antioxidants”

A search of TFEB brought up a 2023 nematode study:

“We searched for effectors acting downstream of the transcription factor EB (TFEB), known as HLH-30 in C. elegans, because TFEB/HLH-30 is necessary across anti-aging interventions. Its overexpression is sufficient to extend C. elegans lifespan, and reduce biomarkers of aging in mammals including humans.

While investigating the potential role of autophagy in hlh-30 dependent longevity of the mxl-3 C. elegans mutant, we found that the current model has exceptions. Contrary to expectation, we found that autophagy is not activated in the mxl-3 mutant, and that neither autophagy nor lysosomal activity are required for the longevity phenotype observed in these mutant animals. mxl-3 longevity is hlh-30-dependent but autophagy-independent.

Instead, we found the gene encoding Alcohol DeHydrogenase ADH-1 induced in mxl-3 and other hlh-30-dependent anti-aging interventions. adh-1 is induced in an hlh-30-dependent manner in longevity models caloric restriction (eat-2), insulin insensitivity (daf-2), and mTOR inhibition (let-363 RNAi).

We present an alcohol-dehydrogenase-mediated anti-aging response (AMAR) that is essential for C. elegans longevity driven by HLH-30 overexpression, caloric restriction, mTOR inhibition, and insulin-signaling deficiency. Overexpression of ADH-1 is sufficient to activate AMAR, which extends healthspan and lifespan by reducing levels of glycerol, an age-associated and aging-promoting alcohol.”

https://www.cell.com/current-biology/fulltext/S0960-9822(23)00128-8 “Increased alcohol dehydrogenase 1 activity promotes longevity” (not freely available) Thanks to Dr. Eyleen O’Rourke for providing a copy.

A 2022 human study found that chronic ADH1 activation occurs in liver disease:

“Activity of total ADH, ADH isoenzymes and aldehyde dehydrogenase (ALDH) was evaluated in the blood serum of patients with primary biliary cholangitis (PBC), a chronic autoimmune disease of the liver. An increase in class I ADH and total ADH activity indicates that the isoenzyme class I ADH is released by compromised liver cells and can be useful diagnostic markers of PBC.”

https://link.springer.com/article/10.1007/s00005-022-00667-4 “An Assessment of the Serum Activity of ADH and ALDH in Patients with Primary Biliary Cholangitis”

Chronically activating any of the body’s systems points to a problem. There’s has to be a balance.

A 2022 rodent study investigated ADH1 activation and MEK1/2 inhibitors for beneficial effects:

“Alcohol is mainly catabolized by class I alcohol dehydrogenase (ADH1) in liver. ADH deficiency can aggravate ethanol-induced tissue injury.

Extracellular signal-regulated kinases 1/2 (ERK1/2) is involved in alcohol metabolism. However, the relationship between ERK1/2 and ADH1 remains unclear.

Mitogen-activated protein kinases 1/2 (MEK1/2) is required to phosphorylate and activate ERK1/2. Protein expression of phosphorylated ERK1/2 in liver is inversely associated with ethanol-induced liver injury and hepatocytes apoptosis, suggesting inhibition of ERK1/2 may protect hepatocytes from ethanol-induced cytotoxicity. We hypothesize that inhibition of ERK1/2 by MEK1/2 inhibitors may protect hepatocytes from ethanol cytotoxicity by activating ADH1 expression.

Results showed MEK1/2 inhibitors significantly increased ADH1 protein expression by inducing its transcription activity. Our findings revealed inhibition of ERK1/2 can significantly increase ADH1 expression, indicating MEK1/2 inhibitors may possess potential application in alcohol-related diseases.”

https://link.springer.com/article/10.1007/s11033-022-07361-w “MEK1/2 inhibitors induce class I alcohol dehydrogenase (ADH1) expression by regulating farnesoid X receptor in hepatic cell lines and C57BL/6J mouse” (not freely available)

Chronically inhibiting any of the body’s systems also points to a problem.

A 2022 rodent study investigated TFEB activation and MEK1/2 inhibitors for beneficial effects:

“Inhibiting MEK/ERK signaling using a clinically available MEK1/2 inhibitor induces protection of neurons through autophagic lysosomal activation mediated by transcription factor EB (TFEB) in a model of AD.”

https://www.nature.com/articles/s41380-022-01713-5 “MEK1/2 inhibition rescues neurodegeneration by TFEB-mediated activation of autophagic lysosomal function in a model of Alzheimer’s Disease”