Genetic factors

Fructose and survival

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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.

rstb20220230f04

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.

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Part 2 of Harnessing endogenous defenses with broccoli sprouts

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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.

ijms-24-13448-g004

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

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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.

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Fermented oats

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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.

nutrients-15-03521-g002-550

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.

Reversing biological age in rats

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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:

second follow-on results

“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

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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

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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.

23 electrophilics

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”

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

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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”

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Neuritogenesis

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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.

experiment design

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:

  1. 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
  2. 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.

j_nipt-2023-0008_fig_002

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

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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.

arthur and robbins cohorts

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.

peaks

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

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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).

insulin insensitivity longevity model

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”

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Adverse Childhood Experiences, Part 2

gettingwell4 2-3 stars Required further work, Brain development, Cerebellum, Cerebrum, Epigenetics, Hippocampus, Limbic system, Lower brain, Memory, Stress , , , , , ,

A request was made to present studies that investigated epigenetic impacts of corporal punishments or physical trauma to children or adolescents. Here’s a follow-on of the 2015 Grokking an Adverse Childhood Experiences (ACE) score, since physical abuse is one factor of an ACE score.

1. The largest problem is that a person filling out an ACE questionnaire or Childhood Trauma Questionnaire can’t provide first-hand answers of their own experiences during womb life, infancy, and early childhood. These critical development periods are more impacted by adversity than are later life windows.

Human brains aren’t developed enough before age 3 to provide retrospective answers using cerebral memories. A self-reported ACE score can’t possibly address what happened during the times when we were most vulnerable to disrupted neurodevelopment. And good luck with parents providing factual histories of whether they physically or emotionally neglected, physically or emotionally abused, or otherwise adversely treated their fetus, infant, and young child.

2. Another problem is researchers can pretty much choose whatever questions they want as input criteria. I’ve seen pliable ACE scores developed from 5- to 25-item questionnaires.

Do these questionnaires cover all relevant adverse childhood experiences? For example, are researchers permitted to use as inputs societal-created adversities a child may have lived through such as the Khmer Rouge or Cultural Revolution? Studies are just starting to investigate adverse childhood experiences created by worldwide abuses of authority since 2020.

3. Other problems were discussed in a 2023 paper https://www.sciencedirect.com/science/article/abs/pii/S0145213423003162 “Adverse childhood experiences and adult outcomes using a causal framework perspective: Challenges and opportunities” (not freely available), two of which were:

  • Adding up ACE factors to a cumulative score ignores the impact of synergistic sets. For example, although both cumulative ACE scores are 2, a child who was physically and sexually abused would probably be more adversely affected than a child whose parents divorced or separated, and also had a family member incarcerated.
  • At any given time point, and especially with older people, there’s a potential selection bias against those most affected by adverse childhood experiences, such as those who died.

Using flawed, squishy, cumulative ACE scores as inputs, here are two 2023 studies that found epigenetic associations:

“We tested the following pre-registered hypotheses: Mothers’ adverse childhood experiences are correlated with DNA methylation (DNAm) in peripheral blood during pregnancy (hypothesis 1) and in cord blood samples from newborn infants (hypothesis 2), and women’s depression and anxiety symptoms during pregnancy mediate the association between mothers’ ACE exposure and prenatal/neonatal DNA methylation (hypothesis 3).

  1. Hypothesis 1: In 896 mother−infant pairs with available methylation and ACE exposure data, there were no significant associations between mothers’ ACE score and DNAm from antenatal peripheral blood, after controlling for covariates.
  2. Hypothesis 2: In infant cord blood, there were 5 CpG sites significantly differentially methylated in relation to mothers’ ACEs (false discovery rate < .05), but only in male offspring. Effect sizes were medium. CpG sites were in genes related to mitochondrial function and neuronal development in the cerebellum.
  3. Hypothesis 3: There was no mediation by maternal anxiety/depression symptoms found between mothers’ ACEs score and DNAm in the significant CpG sites in male cord blood.”

https://www.jaacap.org/article/S0890-8567(23)00313-1/fulltext “Epigenetic Intergenerational Transmission: Mothers’ Adverse Childhood Experiences and DNA Methylation”

“In this study, the effect of cumulative ACEs experienced on human maternal DNAm was estimated while accounting for interaction with domains of ACEs in prenatal peripheral blood mononuclear cell samples. Intergenerational transmission of ACE-associated DNAm was explored used paired maternal and neonatal cord blood samples. Replication in buccal samples was also explored.

We used a four-level categorical indicator variable for ACEs exposure: none (0 ACEs), low (1–3 ACEs), moderate (4–6 ACEs), and high (> 6 ACEs). 🙄

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https://www.researchsquare.com/article/rs-2977515/v1 “Effect of Parental Adverse Childhood Experiences on Intergenerational DNA Methylation Signatures”

Natural ways to modify GDF11

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

Three 2023 studies to follow up mention of GDF11 in the Brain endothelial cells post. Two are selected for non-pharmaceutical interventions people can do on their own. Let’s start with a human exercise study:

“We explored the exercise-related regulation of Growth Differentiation Factor 11 (GDF11) in cerebrospinal fluid (CSF) and blood. Samples of serum, plasma, and CSF were obtained before and 60 min after acute exercise (90 min run) from twenty healthy young individuals. Additional serum and plasma samples were collected immediately after run. GDF11 protein content, body composition, physical fitness, and cognitive functions were evaluated.

Controversies regarding the role of GDF11 in aging originate mainly from the absence of a reliable, validated and widely accepted method of GDF11 detection. To support the reliability of our findings as well as to distinguish GDF11 from its close homologue GDF8, we determined GDF11 in CSF, serum, and plasma, by immunoblotting, using two different GDF11-specific antibodies, as well as GDF11/GDF8 non-specific antibody. These antibodies have been previously successfully used by others. Reliability of our findings is further supported by correlations between GDF11 in serum and plasma, as well as between GDF11 and serum GDF11/GDF8.

We report an association between levels of GDF11 and adiponectin in CSF as well as in serum after acute endurance exercise. These observations support potentially synergic effects of GDF11 and adiponectin on the brain. The experimental design we implement seems to represent a reliable model to study regulation of bioactive molecules, potential mediators of neuroprotective effects of exercise in the human brain.

We show for the first time a direct link between endurance exercise and GDF11 levels in human cerebrospinal fluid. This study provided the first albeit indirect (correlative) evidence on the putative role of GDF11 in promoting healthy aging in humans, by demonstrating a tight relationship between serum GDF11 and peak power output. We extend this observation by showing that the level of physical fitness is an important determinant of regulation of GDF11 by acute exercise.

In this work, we confirm in a bigger cohort our previous finding that blood-brain barrier permeability, as assessed by CSF/serum albumin ratio, is decreased after an acute bout of endurance exercise. We observed a modest positive correlation between CSF/serum albumin ratio and CSF/serum GDF11/GDF8 ratio, with a trend also for GDF11. However, exercise-induced changes of CSF/serum albumin ratio and that of GDF11 or GDF11/GDF8 did not correlate, indicating that there are other factors that could modulate levels of this growth factor rather than blood-brain barrier permeability.”

https://www.frontiersin.org/articles/10.3389/fendo.2023.1137048/full “Acute endurance exercise modulates growth differentiation factor 11 in cerebrospinal fluid of healthy young adults”

Next is a rodent study of intermittent fasting before and after cerebral ischemia:

“The present study focused on the cerebral angiogenesis effect of intermittent fasting (IF) on ischemic rats. Rats were fed within strict time periods for 8 h out of every 24 h, with free access to food between 0800 and 1600 h.

In the first step, we designed different time schedules (10 d, 1 month, and 3 months) of IF before middle cerebral artery occlusion (MCAO). We monitored whether IF accelerated neurobehavioral recovery and induced expression of endothelial cells after MCAO. Then we explored whether GDF11 and downstream signals mediated angiogenesis in the peri-infarct area.

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We found that 3 months (p < 0.01) and 1 month (p < 0.05) of IF conditioning, respectively, markedly increased GDF11-positive cells in the peri-infarct area 3 d after MCAO compared with ad libitum dietary regimen. There were no significant differences between the cerebral ischemia (CI) + ad libitum group and the CI + IF 10-day group.

We also assayed plasma expression pattern of GDF11 protein. Plasma level of GDF11 protein was significantly upregulated in the IF dietary groups compared with the ad libitum dietary group 3 d after MCAO, which was consistent with the brain level. However, short-term CI + IF 10-day group results were not statistically different from CI + ad libitum group.

Taken together, our results strongly indicated that pretreatment of long-term IF might promote circulation of GDF11 and cerebral GDF11 protein during the post-ischemic, recovery period. Preoperative long-term IF might be beneficial for inducing cerebral angiogenesis in acute cerebral infarction.

These findings suggested that the longer the period of IF before MCAO, the better the protective effects after surgery.”

https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0282338 “Long-term intermittent fasting improves neurological function by promoting angiogenesis after cerebral ischemia via growth differentiation factor 11 signaling activation”

Per Week 28 of Changing to a youthful phenotype with broccoli sprouts, using species maximum lifespan to estimate a human-equivalent multiplication factor that can be applied to a rat post-development time period is 122.5 years / 3.8 years = 32.2. Applying it to this study’s findings:

  • 10 rat days (322 human days) of intermittent fasting provided little protection from cerebral ischemia;
  • 1 rat month (32.2 human months) of intermittent fasting had better protection; and
  • 3 rat months (a little over 8 human years) of intermittent fasting had even stronger protection.

Is it worth the hassle of intermittently fasting every day for years to prevent a future stroke, or better recover from one, or keep other subclinical / clinical diseases from accelerating, or keep aging from accelerating? This study also cited more immediate benefits of intermittent fasting.

Might be too late for a gradual approach for people who are already diseased or close, though, like subjects in this human study:

“We aimed to explore the correlation among serum GDF11, the severity of coronary artery lesions, and the prognosis of patients with ST-segment elevation myocardial infarction (STEMI). A total of 367 patients were enrolled and divided into control (n = 172) and STEMI (n = 195) groups. Control group fulfilled the following criteria:

  1. Presented with typical chest tightness, chest pain, or other discomfort symptoms on admission;
  2. Electrocardiogram examination suggested ST-T changes;
  3. Levels of myocardial injury markers did not suggest abnormalities; and
  4. The diagnosis of unstable angina was considered clinically valid.

14 variables that were significant in univariate logistic regression analysis were included in the subsequent multivariate logistic regression analysis. Multivariate analysis indicated that smoking, diabetes, C-reactive protein, homocysteine, and lipoprotein (a) were positively correlated with STEMI occurrence, whereas serum GDF11 and the Apolipoprotein A1-to-Apolipoprotein B ratio were negatively correlated with STEMI occurrence.

Serum GDF11 was negatively correlated with severity of coronary lesions, and was also an independent prognostic indicator of major adverse cardiovascular events in patients with STEMI.”

https://link.springer.com/article/10.1007/s12265-023-10358-w “Correlation Between GDF11 Serum Levels, Severity of Coronary Artery Lesions, and the Prognosis of Patients with ST-segment Elevation Myocardial Infarction” (not freely available)

A flawed broccoli sprouts clinical trial

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

This 2023 human study investigated commercially available broccoli sprouts’ effects on platelets. I’ll provide details of some procedures, but not of findings, as there were several issues:

“Administration of intervention (sulforaphane/placebo) was followed in 90 min by administration of standardized caloric challenge PhenFlex. Urine samples were classified into three groups: (A) baseline, green line, (B) after intervention or placebo, blue lines, and (C) after PhenFlex challenge, red lines. Samples were divided into 5 timepoints: (0) baseline, (1) <60 min after intervention or placebo, (2) >60 min after intervention or placebo, (3) <60 min after PhenFlex challenge, and (4) >60 min after PhenFlex challenge.

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Shortly (maximum of 3 min) before administration, sprouts were cut approximately 1 cm below the leaves, weighed, and mashed with a small amount of tap water (approximately 13°C) in a kitchen blender for 30s at room temperature. Subsequently, tap water was added to a total amount of 250 mL and participants were instructed to drink the entire mixture.

Commercially available pea sprouts (Affilla Cress®) were used as placebo in this study since pea sprouts do not contain glucoraphanin/sulforaphane. Affilla Cress (16 g) was prepared and administered in a similar fashion. Blinding of participants was ensured by the even appearance of both drinks and the use of nasal plugs during consumption of the investigational products. 🙂

Ninety minutes after administration of investigational products, participants were asked to drink PhenFlex, a high-fat, high-glucose, high-caloric product. PhenFlex mixtures were freshly prepared, and participants were instructed to consume the drink within 5 min.”

https://www.frontiersin.org/articles/10.3389/fnut.2023.1204561/full “The beneficial effect of sulforaphane on platelet responsiveness during caloric load: a single-intake, double-blind, placebo-controlled, crossover trial in healthy participants”

Two main issues were:

1. It was stated throughout that sulforaphane did or could do this and that. None of this was supported by sulforaphane intake measurements / estimates, although measuring equipment was available.

Researchers couldn’t assume that blending 16 grams of broccoli sprouts of unknown age creates x amount of sulforaphane. 3-day-old broccoli sprouts have the optimal yields measured 6 broccoli varieties’ sulforaphane content over 3, 5, and 7-day ages, and published 15 different answers.

Sulforaphane and two metabolites’ urinary output was measured. Supposing that only output measurements were adequate leads to the second main issue.

2. Genes were asserted for certain effects. Plausible alternate explanations such as individual differences in gut microbiota composition, excretion, and metabolism weren’t explored.

These researchers knew or should have known about the 2016 https://onlinelibrary.wiley.com/doi/abs/10.1002/mnfr.201600766 “Stabilized Sulforaphane for Clinical Use: Phytochemical Delivery Efficiency” (not freely available). That study measured two known sulforaphane inputs, and in ten people each, blood plasma and urinary outputs.

The first sulforaphane input had sulforaphane bioavailability from 19.5% to 86.9% of dose. The second input ranged from 48% to 96% of dose. Widely different responses to sulforaphane intake prompted those researchers to state:

“These differences in SF bioavailability may be due to differences in gut microbial metabolism, in the levels of drug metabolizing enzymes (e.g. well-known polymorphisms of glutathione S-transferases that catalyze the conjugation of SF with glutathione), and in excretion kinetics. Innate metabolic differences must not be discounted when assessing the metabolism of SF.”

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The brain-gut-lung circuit

gettingwell4 4-5 stars Advanced knowledge, Dopamine, Epigenetics, Glutamate, Immune system, Neurochemicals, Serotonin, Stress , ,

This 2023 rodent study investigated mechanisms of improving stress-worsened respiratory viral infection:

“Our study demonstrates that chronic psychological stress significantly increases host vulnerability to influenza A virus (IAV) infection characterized by a distorted gut microbiome and deregulated alveolar macrophages (AMs) response. We show that microbiome-derived γ-aminobutyric acid (GABA) functions as a tonic signal to support survival, self-renewing, and immunoregulation of AMs, and hence optimized pulmonary defensive response.

Chronic psychological stress causes gut microbiome dysbiosis and defective GABA generation, leading to loss of AMs homeostasis and aggravated viral pneumonia. The data indicate that:

  1. Microbial GABA is released in the circulation,
  2. Sensed by AMs via the GABAA receptor,
  3. Promoting cellular mitochondrial metabolism,
  4. For increased production of α-ketoglutarate (αKG),
  5. Which triggers Tet2-mediated DNA hydroxymethylation,
  6. To enable PPARγ-centered gene program,
  7. Supporting AMs homeostasis and function.

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  • Re-localization of GABA-generating probiotics,
  • Supplementation of αKG, or
  • Adoptive transfer of GABA-conditioned macrophages,
  • Substantially rectifies stress-induced disruption inter-organ communication, and
  • Alleviates symptoms of viral pneumonia.

Our current study unveils an unappreciated regulatory circuitry that connects the brain, gut, and lung to mediate neurological modulation of host defensive response.”

https://www.sciencedirect.com/science/article/pii/S2090123223001716 “Gut microbial GABAergic signaling improves stress-associated innate immunity to respiratory viral infection”

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