Year Three of Changing to a youthful phenotype with sprouts

March 25, 2023June 13, 2023 gettingwell4 4-5 stars Advanced knowledge, Epigenetics, Immune system, Stress

1. I’ve continued daily practices from Year Two with microwaved 3-day-old broccoli, red cabbage, and mustard sprouts for 13 times longer now than any sulforaphane clinical trial. I haven’t had another three-year period in my life where I wasn’t sick even once!

The main difference over the past year is that I eat AGE-less chicken vegetable soup 3-4 times a week rather than twice a day. That was just too boring, plus I stopped eating lunch. My other dinners are often steamed vegetables and seafood.

2. I frame the above and following tactics as part of a strategy of delaying evolutionarily-determined programs from doing more to kill my my body every year:

Eat 16-hour sprouted Avena nuda oats for breakfast to give my trillion+ gut microbiota what they need;
Eat 3-day-old hulled Avena sativa oat sprouts twice a day to provide antioxidants in another form, avenanthramides;
Eat AGE-less chicken vegetable soup 3-4 times a week to prevent diet-induced problems;
Take supplements that promote healthspan twice a day;
Exercise at least 30 minutes daily;
Take yeast cell wall β-glucan daily, with nothing else an hour before or after; and
Avoid undue stress by working from home 40 hours a week in my 26^th year as a professional software developer.

These tactics’ main components activate AMPK, Nrf2, and associated signaling pathways, and inhibit pro-inflammatory pathways such as NF-κB.

But fixing inflammation doesn’t repair all existing damage. I wonder what could have been physically resolved if I had started thirty+ years earlier.

3. One place I take clues from are successful anti-aging animal research efforts such as a study reviewed here earlier this month. Last curated in Improve your internal environment, improve its constituents’ functions, it used plasma fraction treatment. Plasma fraction eclipsed a caloric restriction treatment’s previous record for maximum species lifespan by 5%.

This type of research clearly isn’t a priority for official sponsors to fund, though. Take responsibility for your own one precious life.

Eat broccoli sprouts to protect your lungs

March 19, 2023March 19, 2023 gettingwell4 4-5 stars Advanced knowledge, Epigenetics, Immune system, Stress Control group

This 2023 human cell study investigated sulforaphane’s effects on tuberculosis infections:

“Basic research efforts on tuberculosis (TB) immunotherapy are currently only the tip of the iceberg. This study highlights the association between autophagy-related genes and immune infiltration in TB, an infectious pathogen that has been around for tens of thousands of years.

Sulforaphane (SFN) is readily absorbed into the bloodstream by the intestine due to its lipophilic nature. Experiments in this study TB patient cells showed that SFN could promote autophagy in macrophages infected with Mycobacterium abscessus (Mab). Intracellular bacterial load of macrophages was associated with SFN-enhanced cellular autophagic processes.

The relationship between autophagy and immune cells is complex, and recurrence of tuberculosis is significantly influenced by intracellular mycobacteria of macrophages. Macrophages have longer lifespans than neutrophils, and provide shelter for mycobacteria as they are better suited than neutrophils to establish strategies for targeting autophagy.

This is one of the reasons why autophagy in macrophages was the focus of this study. Appropriate autophagy is beneficial for the body and controls Mtb replication, but autophagic programmed cell death can activate tissues to produce an excessive inflammatory response, resulting in severe damage to lung tissues.

Autophagy-related genes regulated by SFN have good diagnostic potential, with FOXO1 potentially serving as a target for TB immunotherapy. Downstream targets of FOXO1 include important pro-inflammatory signaling molecules such as IL-1β and TNF-α, which are important for control of mycobacterium.”

https://www.sciencedirect.com/science/article/pii/S156757692300276X “Identifying autophagy-related genes as potential targets for immunotherapy in tuberculosis”

Take yeast cell wall β-glucan, too, and train your immune system.

Does eating broccoli sprouts influence biological age?

March 18, 2023March 19, 2023 gettingwell4 2-3 stars Required further work, Epigenetics, Stress Control group, Embryo development, Genetic factors

A 2023 review of 28 human clinical trials investigating broccoli sprout compounds brought up this post’s title by omitting discussion of it:

“In order to determine the effective reference dose of a broccoli sprouts beverage for detoxifying carcinogenic air pollutants (benzene), Chen et al. administrated a drink enriched with glucoraphanin (GR) and sulforaphane (SFN) from 3-day-old broccoli sprouts to healthy adults. Researchers focused on excretion of metabolites SFN-NAC, SFN-CYS, and non-esterified SFN, which represent 80–81%, 12–14%, and 5–7% of total SFN forms, respectively.

Excretion percentage did not change during the intervention, indicating that bioavailability remained constant.

Enhanced excretion of the urinary biomarker of benzene detoxification S-phenylmercapturic acid (SPMA) was measured in urine collected every 12 h during the 10-day intervention. Out of 132 samples analyzed, >95% had detectable concentrations of SPMA, being significantly increased after consumption of the high dose of beverage (600 and 40 μmol GR and SFN, correspondingly), suggesting that consumption of >10 μmol SFN per 24 h may represent the lowest effective dose of the BSE affecting this biomarker.“

https://www.mdpi.com/2072-6643/15/6/1424 “Systematic Review on the Metabolic Interest of Glucosinolates and Their Bioactive Derivatives for Human Health”

These reviewers did much hand waving to draw their conclusions. They ignored that the only way randomized trials become better than non-randomized trials is in dealing with confounders.

The largest confounder with glucoraphanin is that an individual’s gut microbiota, not their human cells, metabolize it into isothiocyanates. A glucoraphanin randomized trial has to have sufficient numbers of subjects in each group to adequately deal with confounding individual differences in gut microbiota.

I highlighted the largest of the 28 trials:

Sulforaphane studies have fewer confounders. Even so, Upgrade your brain’s switchboard with broccoli sprouts stated:

“Power analysis calculations suggest that a sample size of n = 50 would yield a significant result.”

An insufficient number of subjects in both the half dose and full dose groups caused that study’s researchers to frame their results as “suggesting that consumption of >10 μmol SFN per 24 h may..” rather than asserting significant results.

Addressing this post’s title, it’s been ten years since epigenetic clocks came into use. This review highlighted by omission that there still hasn’t been even one investigation of isothiocyanates’ effects on human biological age as measured by epigenetic clocks.

A 40 μmol ≈ 7 mg sulforaphane “high” dose of the cited study is easily achievable with microwaved 3-day-old broccoli sprouts. There’s little question that healthy people activating AMPK, Nrf2, and associated signaling pathways, and inhibiting pro-inflammatory pathways such as NF-κB with sulforaphane, will experience beneficial effects.

The cited study found no change in sulforaphane treatment bioavailability over ten days, and a predecessor study found the same over 12 weeks. I’ll guess those bioavailability findings will extend over longer time periods.

Where are the researchers who will take the next step to show isothiocyanate treatments cause positive changes in epigenetic clock / biological age measurements?

Eat broccoli sprouts for depression, Part 3

March 13, 2023March 16, 2023 gettingwell4 4-5 stars Advanced knowledge, Cerebrum, Epigenetics, Hippocampus, Hypothalamus, Limbic system, Memory, Stress Brain neurons, Control group, Critical period, Genetic factors, Neural plasticity, Prefrontal cortex

Here are two papers published after Part 2 that cited the Part 1 rodent study, starting with a 2023 rodent study performed by several Part 1 coauthors:

“We used a low-dose LPS-induced endotoxaemia model to mimic clinical characteristics of sepsis. We found that adolescent LPS treatment was sufficient to increase levels of inflammatory factor TNF-α in both the medial prefrontal cortex (mPFC) and hippocampus at post-natal day P22.

P21 LPS-treated mice were injected with sulforaphane (SFN) or saline intraperitoneally at P49 and then subjected to subthreshold social defeat stress (SSDS). We found that SFN preventative treatment significantly:

Decreased the social avoidance, anhedonia, and behavioural despair detected by the social interaction test, sucrose preference test, tail suspension test, and forced swim test, respectively.

Decreased anxiety-like behaviours without affecting locomotor activities.

Increased Nrf2 and brain-derived neurotrophic factor (BDNF) levels in the mPFC of P21 LPS-treated mice after SSDS compared with saline control mice.

The above results suggest that activation of the Nrf2-BDNF signalling pathway prevents the effect of adolescent LPS-induced endotoxaemia on stress vulnerability during adulthood.

These results suggest that early adolescence is a critical period during which inflammation can promote stress vulnerability during adulthood. This might be due to increased inflammatory response in the mPFC, and mediated by decreased levels of Nrf2 and BDNF. These findings may shed light on the potential use of SFN as an alternative preventative intervention for inflammation-induced stress vulnerability.”

https://link.springer.com/article/10.1007/s00213-022-06285-4 “Lipopolysaccharide-induced endotoxaemia during adolescence promotes stress vulnerability in adult mice via deregulation of nuclear factor erythroid 2-related factor 2 in the medial prefrontal cortex” (not freely available)

This study demonstrated that adolescent diseases and stresses don’t necessarily develop into adult social problems. A timely intervention may even prevent future adult problems.

The one-time 10 mg/kg sulforaphane dose was the same as Part 1’s dose, a human equivalent of which is (10 mg x .081) x 70 kg = 57 mg.

Per Human relevance of rodent sulforaphane studies, that’s above a tolerable oral dose of ≈ 50 mg in one serving.
I didn’t have a problem with eating half that twice a day via microwaved broccoli sprouts from Week 6 through Week 56.

I’d like to know more about how subjects’ memories of adverse events were retained, and subsequently affected their biology and behavior. Pretty sure limbic structures like the hypothalamus as well as lower brain structures played a part.

A 2022 review summarized what was known up to that time regarding Nrf2 and depression:

“Sulforaphane, an organosulfur compound isolated from Brassicaceae plants, is a potent natural NRF2 activator. Sulforaphane:

Exerts antidepressant- and anxiolytic-like activities and inhibits HPA axis and inflammatory response.

Has both therapeutic and prophylactic effects on inflammation-related depression.

Confers stress resilience.

Protects neurons via autophagy and promotes mitochondrial biogenesis by activating Nrf2.”

https://www.sciencedirect.com/science/article/pii/S2213231722002944 “Nrf2: An all-rounder in depression”

Oat β-glucan effects

March 11, 2023March 12, 2023 gettingwell4 4-5 stars Advanced knowledge, Epigenetics, Immune system Control group, Genetic factors

Three papers on oat β-glucan’s effects in humans, starting with a 2023 study that compared different doses:

“Two randomized, double-blind, controlled studies were conducted with asymptomatic subjects between 20 and 40 years of age, male or female, normal weight or overweight.

In the first study – a crossover trial comprising two days of testing (β-glucan and control) separated by at least one week – 14 subjects ingested a breakfast with or without β-glucan from oats (5.2 g). Results indicate that acute intake of 5 g β-glucan slows transit time and decreases hunger sensation and postprandial glycaemia without affecting bile-acid synthesis. These changes were associated with decreased plasma insulin, C-peptide, and ghrelin, and increased plasma gastric inhibitory polypeptide and pancreatic polypeptide.

In the second study, 32 subjects were distributed into 2 groups to ingest daily foods with (3 g/day) or without β-glucan for 3 weeks. Results indicate a regular daily intake of 3 g β-glucan is not sufficient to have an effect on fecal microbiota composition, suggesting that health-promoting effects at 3 g/d are probably due more to their physiological effect in the proximal part of the gastrointestinal tract than to their prebiotic effect in the colon.”

https://www.mdpi.com/2304-8158/12/4/700 “Modulation of Postprandial Plasma Concentrations of Digestive Hormones and Gut Microbiota by Foods Containing Oat ß-Glucans in Healthy Volunteers”

I’ll use a 2021 study Rapid Determination of β-Glucan Content of Hulled and Naked Oats Using near Infrared Spectroscopy Combined with Chemometrics to estimate my daily β-glucan intake. Those researchers tested 100 varieties of Avena nuda that varied between 3.12% and 5.22% β-glucan. My intake from 82 g (dry weight) of hulless oats (cinnamon sprinkled for taste) is probably between (82 g x .0312) = 3 g and (82 g x .0522) = 4 g.

They also tested 79 varieties of hulled Avena sativa that varied between 3.1% and 5.5% β-glucan. Oat sprouts analysis tested a Avena sativa variety where the β-glucan content decreased from 3.48% to 2.10% over four days of sprouting, a 40% reduction.

My daily β-glucan intake from 40 g (dry weight) of three-day-old hulled oat sprouts is probably 1 g [(40 g x .031) x .6 = 1 g and (40 g x .055) x .6 = 1 g]. That’s okay, because oat sprouts have other benefits per Oat sprouts analysis and Advantages of 3-day-old oat sprouts over oat grains.

My daily oat β-glucan intake is 4 – 5 grams. I’ve maintained that for two years, and don’t see any reason to stop.

A second 2023 paper from a clinical trial investigated effects of combining oat bran along with orange juice:

“Orange juice (OJ) is a rich dietary source of bioactive flavanones, and consuming OJ has been associated with beneficial effects including decreased inflammation and improved lipid profiles. However, dietary recommendations are to limit OJ consumption to one serving per day due to high sugar and low fiber content. Metabolic concerns are increased postprandial insulin response to a high sugar load which in individuals at risk may promote insulin resistance.

Consumption of 22 g oat bran containing 6 g of β-glucan together with 500 mL of OJ by healthy subjects impacts on OJ flavanone bioavailability with the 0-24 post-intake excretion of phase II metabolites, such as hesperetin-7-glucuronide, being reduced ~3-fold. This was not a consequence of bran affecting the rate of gastric transport, and underlying mechanisms behind reduced excretion of OJ flavanone metabolites remain a matter of conjecture.

The pool of bound phenolics in bran linked to polysaccharides appears not to be converted to free phenolics. It was rather principally a consequence of a bran-mediated increase in quantities of flavanones passing from the upper to the lower bowel where they were subjected to microbiota-mediated catabolism.”

https://www.sciencedirect.com/science/article/abs/pii/S0891584923000515 “Bioavailability of orange juice (poly)phenols: β-glucan-rich oat bran decreases urinary excretion of flavanone phase II metabolites and enhances excretion of microbiota-derived phenolic catabolites” (not freely available) Thanks to Dr. José Manuel Moreno-Rojas for providing a copy.

This paper referenced a preliminary study by many of the same coauthors that found oat bran with 3 g of β-glucan didn’t have similar effects.

A 2022 meta-analysis investigated differences between whole oats and purified β-glucan:

“This systematic review and meta-analysis evaluated the impact of oats or β-glucan supplements on the lipid profile. Our findings show that both oat and isolated β-glucan interventions can improve lipid profiles, specifically total cholesterol and low density lipoprotein cholesterol (LDL) concentrations, and should be incorporated into one’s regular eating habits.

Interventions ranged from 14 to 84 days in length. Quantity of β-glucan ingested (oats and isolated β-glucan) ranged from 1.2 g/day to 11.2 g/day.

Limitations and additional considerations include:

We did not have enough studies that matched total fiber intake between intervention and control groups, and so could not evaluate if results were exclusively influenced by oat/isolated β-glucan supplementation, or if other types of dietary fiber would have a similar impact on lipidemia.

Mechanisms of changes in concentrations of triglycerides (TG) are linked to carbohydrates. An increase in availability of glucose in serum, resulting from absorption of carbohydrates, stimulates secretion of insulin and, as a result, synthesis of fatty acids in the liver is increased. Mixed results found in this and other meta-analyses regarding TG may be related to the fact that oats and isolated β-glucan were frequently administered through day-to-day processed foods which have sugar and other types of refined flour in their recipes.

Different oat cooking procedures, processing methods, and molecular weights modify viscosity and impact in cholesterol concentrations differently. Less processed oats appear to be more effective than processed oat products in improving lipidemia. Higher molecular weight is associated with increased viscosity, and greater reduction in LDL. Also, the process used to treat oats affects its molecular weight, and the highest viscosities were observed as a consequence of dry processes in comparison to ones that exhibit enzymatic activity.

Reducing saturated fat intake may be, in combination with increased viscous fiber intake from oats or isolated β-glucan, the most effective way to improve dyslipidemia. In future studies, amount and type of fat in diet should be evaluated and considered accordingly.”

http://dx.doi.org/10.1016/j.clnesp.2022.12.019 “The separate effects of whole oats and isolated beta-glucan on lipid profile: A systematic review and meta-analysis of randomized controlled trials” (not freely available)

Peripheral vs. brain epigenetic measurements

March 5, 2023May 26, 2023 gettingwell4 2-3 stars Required further work, Epigenetics, Immune system Brain neurons, Control group, DNA methylation, Prefrontal cortex

This 2023 human study investigated associations of peripheral and brain epigenetic measurements:

“Evaluating DNA methylation of brain tissue is challenging owing to the issue of tissue specificity. Consequently, peripheral surrogate tissues are used, resulting in limited progress compared with other epigenetic studies.

Averaging data for each CpG across individuals, saliva–brain correlation (r = 0.90) was higher than that for blood–brain (r = 0.87) and buccal–brain (r = 0.88) comparisons. Among individual CpGs, blood had the highest proportion of CpGs correlated to the brain at nominally significant levels (19.0%), followed by saliva (14.4%) and buccal (9.8%). However, cross-database correlations of correlation coefficients revealed relatively low brain vs. blood: r = 0.27, saliva: r = 0.18, and buccal: r = 0.24.

The majority of methylation in the brain is most likely not synchronized with methylation in the periphery. Despite this, variable CpGs that correlate in the brain and periphery, although in small numbers, may have biological relevance, and could be useful for inferring brain methylation from peripheral tissues.

This study has six major limitations.”

https://www.nature.com/articles/s41398-023-02370-0 “Cross-tissue correlations of genome-wide DNA methylation in Japanese live human brain and blood, saliva, and buccal epithelial tissues”

Real science is messy. Hypotheses are experimentally reevaluated many, many times under varying conditions. I skip over studies where researchers don’t provide meaningful limitation clauses.

Because human nature doesn’t change

March 3, 2023 gettingwell4 Just me

Catching up with Martin Armstrong:

“Nero’s reputation has been the consequence of ancient fake news that to this day, distorts the man and what he stood against for his attempt to drain the swamp.

The Deep State conspired to kill Nero for his reforms, reflecting the growing discontent among the ruling class of the Roman state with Nero’s increasing attack upon corruption. Some wished to replace Nero with a better emperor who understood the Deep State was off limits. Others wished to be free of emperors altogether, and restore a purely Republican form of government with all its free-wheeling corruption.”

Nero the Antichrist – Deep State & Fake News

“The question of TIME has puzzled humanity for millennia. What is it? Does it flow like a river?

Cycles exist both on a fixed level of time as well as a dynamic level within TIME – Longitudinal & Transverse.

Legends of massive waves that drag ships to their doom have often been attributed to giant monsters of something supernatural. They can be explained as cyclical convergences whereby numerous cyclical waves combine together and produce an abnormally large wave that causes amplitudes of individual waves to blend together, producing a huge event.”

How to Use the Forecasting Arrays