Epigenetics

Eat broccoli sprouts instead of antibiotics

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

This 2020 cell study investigated antibiotic effects of broccoli sprout compounds:

“In this work, we asked whether isothiocyanates (ITCs) could act synergistically with each other to increase antibacterial effect. A set of aliphatic ITCs, such as iberin, iberverin, alyssin, erucin, sulforaphene, erysolin, and cheirolin was tested in combination with sulforaphane against E. coli.

All tested ITCs exhibit strong antimicrobial effect individually. Synergistic action observed for iberin, iberverin, and alyssin led to minimal inhibitory concentration necessary for antibacterial effect four- to eight-fold lower than for individual ITCs.

Effectiveness of antimicrobial effect is correlated with both type of ITC used and bacterial growth conditions. The combination of several fold lower concentration of ITCs gives a similar effect as much higher amounts of individual ITCs.

Antimicrobial action of sulforaphane analogs was impaired by specific amino acids. Antibacterial effect of ITC treatment is related to stringent response induction, which is triggered by amino acid starvation.

The use of ITCs as antibacterial agents can be advantageous, as there are very few examples of bacterial resistance to these compounds.”

https://www.frontiersin.org/articles/10.3389/fmicb.2020.591802/full “Induction of the Stringent Response Underlies the Antimicrobial Action of Aliphatic Isothiocyanates”

One of this study’s references was the 2016 Relationship between Chemical Structure and Antimicrobial Activities of Isothiocyanates from Cruciferous Vegetables against Oral Pathogens which found that broccoli and red cabbage compound indole-3-carbinol and mustard compound benzyl isothiocyanate were even more potent antibiotics than half of the aliphatic isothiocyanates in this study:

antibiotic isothiocyanates

Our ancestors evolved to deal with everyday bacteria, viruses, and other pathogens. Not sure about the current virus developed to herd humans into an agenda.

Train your immune system every day! disclosed that I was in Milan, Italy on the same February 22-23, 2020 weekend that ten towns were closed south of Milan. I’ve never experienced any symptoms.

  • One factor in immune response was that fifteen years previous, I’d taken daily steps with yeast cell wall β-glucan to guard against the phenotypical immune system collapse of old age.
  • Another factor was that I’d ridden the filthy Washington DC Metro twice a day to-and-from work for years, and had already been exposed to who knows what.

Treat your gut microbiota well. Give them what they want – including cruciferous sprouts – instead of prescription antibiotics, and expect reciprocity.

Eat broccoli sprouts daily, and manage weight

gettingwell4 2-3 stars Required further work, Epigenetics

This 2018 human study found:

“The objective of this study was to determine whether daily broccoli consumption alters absorption and metabolism of isothiocyanates derived from broccoli glucosinolates. We conducted a randomised cross-over human study (n = 18) balanced for BMI and glutathione S-transferase μ 1 (GSTM1) genotype in which subjects consumed a control diet with no broccoli (NB) for 16 d or the same diet with 200 g of cooked broccoli and 20 g of raw daikon radish daily for 15 d (daily broccoli, DB) and 100 g of broccoli and 10 g of daikon radish on day 16.

On day 17, all subjects consumed a meal of 200 g of broccoli and 20 g of daikon radish. Plasma and urine were collected for 24 h and analysed for sulphoraphane (SF) and metabolites of SF and erucin (ER). (a) BMI < 26 (b) BMI > 26.

sulforaphane and erucin metabolites

db-nb

Plasma AUC [area under the curve] and urinary excretion rates were higher on DB diet than on NB diet. Daily consumption of broccoli interacted with BMI to affect plasma concentrations and urinary excretion of glucosinolate-derived compounds.

Plasma and urinary levels of SF and mercapturic acid pathway products of SF and ER following a broccoli challenge meal were altered when preceded by 16 d of daily broccoli ingestion, and the effect depended on BMI.”

https://www.cambridge.org/core/journals/british-journal-of-nutrition/article/absorption-and-metabolism-of-isothiocyanates-formed-from-broccoli-glucosinolates-effects-of-bmi-and-daily-consumption-in-a-randomised-clinical-trial/ “Absorption and metabolism of isothiocyanates formed from broccoli glucosinolates: effects of BMI and daily consumption in a randomised clinical trial”

Humans are the same, yet we’re each individually unique. These researchers could have explored individual differences, but that wasn’t part of this study’s design.

So we’re left with BMI as a discriminator. I don’t think that’s evidentiarily sufficient.

Eat broccoli sprouts every day. You’ll figure it out.

Red cabbage effects on gut microbiota

gettingwell4 5+ stars Premium, Epigenetics

A tremendous 2021 study involving the group who published Our model clinical trial for Changing to a youthful phenotype with broccoli sprouts:

“The aim was to evaluate the influence of red cabbage extracts on bioaccessibility of their isothiocyanates, and their effect on intestinal microbiota using a dynamic model of human digestion treated with the gut microbiome of obese adults.

Plant plasma membrane vesicles as delivery systems for bioactive compounds has been studied. Diverse types of plant membrane vesicles could be good candidates for this purpose, such as extracellular vesicles, which are spheroids of cytosolic material surrounded by a lipid bilayer, or extracted plasma membrane from fresh plant tissue.

As an example of the latter, we used cauliflower plasma membrane vesicles, which are proteoliposomes with a high proportion of unsaturated fatty acids. There could be an interaction between plant aquaporins found in our vesicles and isothiocyanates present in red cabbage aqueous extract, which could have increased stability.

Plasma membrane vesicles may act as stabilizing carriers and feeding agents for enzymes and bile salts rather than an encapsulating agent per se. However, this aspect should be further studied.

red cabbage sfn, i3c, iberin

In the transversal colon reactor, butyric acid production by gut microbiota had a 3-fold increase after 14-day treatment for free red cabbage aqueous extract when compared to stabilization period. A 3.5-fold increase was observed when using nanonencapsulated extract.

Regarding the descending colon, a 2-fold increase in butyric acid was produced after 14 days of treatment with free red cabbage aqueous extract. A 4-fold increase was observed in production after treatment with nanoencapsulated extract.

Propionic and acetic acids were studied, but no changes were observed. The fact that encapsulated red cabbage extract provided a higher production of butyric acid pointed to future developments for design of a functional ingredient or food product for management of overweightness and obesity.”

https://www.mdpi.com/2304-8158/10/5/1038/htm “The Influence of Red Cabbage Extract Nanoencapsulated with Brassica Plasma Membrane Vesicles on the Gut Microbiome of Obese Volunteers”

This study demonstrated that iberin was initially the third highest isothiocyanate of red cabbage after glucosinolate hydrolysis. Iberin surpassed sulforaphane to become the predominant isothiocyanate – in both free and nanoencapsulated forms – when it reached the lower colon, where most of our gut microbiota reside.

These in vitro findings were after 14 days, though, which doesn’t happen in healthy humans in vivo. Also, if sulforaphane metabolites such as dithiocarbamates and I3C breakdown products such as DIM were measured, these findings may have changed.

As noted in Tailoring measurements for broccoli sprouts, study findings of mature plants don’t necessarily apply to their sprouts. Lab analyses of broccoli sprout compounds used 9-day-old red cabbage sprouts to measure iberin (3MSOP-ITC in Figure 5). Haven’t found recent studies on iberin’s effects on gut microbiota and intestinal epithelial cells.

This study showed “a 3 to 4-fold increase in production of butyric acid with encapsulated extract treatment.” Keep leading the way. 🙂

Eat oats and regain cognitive normalcy

gettingwell4 5+ stars Premium, Epigenetics, Hippocampus, Hypothalamus, Immune system, Limbic system, Memory, Stress , , ,

This 2020 rodent study investigated effects of different diets:

“The present study aimed to evaluate effects of β-glucan on the microbiota gut-brain axis and cognitive function in an obese mouse model induced by a high-fat and fiber-deficient diet (HFFD). After long-term supplementation for 15 weeks, β-glucan prevented HFFD-induced cognitive impairment, assessed behaviorally by object location, novel object recognition, and nesting building tests:

  • Long-term β-glucan supplementation suppressed microglia activation and inflammation in hippocampus of HFFD-fed mice;
  • β-glucan attenuated deleterious engulfment of synapses by activation of microglia seen in HFFD mice;
  • β-glucan significantly prevented upregulation of TNF-α, IL-1β, and IL-6 mRNA expression in hippocampus; and
  • A broad-spectrum antibiotic intervention abrogated β-glucan-induced improvement in cognitive function, highlighting the essential role of gut microbiota to mediate cognitive function and behavior.

We found that short-term β-glucan supplementation did not change cognitive behavior in HFFD fed mice. HFFD feeding for 7 days dramatically changed gut microbial profile, with β-glucan-fed mice clustered apart from HFFD-fed mice sample, suggesting:

  • Quick changes in gut microbiota are induced by short-term β-glucan consumption and
  • Possible causality of gut microbiota profile on cognition.

7% β-glucan 7% nondigestible fiber

β-glucan supplementation increased place discrimination ratio in object location test compared with HFFD mice; however, there was no significant difference in total exploration time with objects during test phases between the two groups. Higher place discrimination index in β-glucan supplementation group was not due to better general performance, but increased recognition memory.

Results provide consistent evidence linking increased β-glucan intake to improved:

  • Gut microbiota profile;
  • Intestinal barrier function;
  • Reduced endotoxemia; and
  • Enhanced cognitive function via more optimized synaptic and signaling pathways in critical brain areas.

It is speculative that β-glucan improvement of gut microbiota composition, but not necessarily diversity per se, may be most critical for improved cognition. Enhanced consumption of β-glucan-rich foods is an easily implementable nutritional strategy to attenuate diet-induced cognitive decline.”

https://microbiomejournal.biomedcentral.com/articles/10.1186/s40168-020-00920-y “β-glucan attenuates cognitive impairment via the gut-brain axis in diet-induced obese mice”

This study did well by elaborating It’s the fiber, not the fat and Eat oats to prevent diabetes related findings. How many humans eat themselves into essentially the same situation as this HFFD group with no gut-microbiota-friendly dietary fiber?

Experiments were with β-glucan 1,3/1,4 found in oats. β-glucan 1,3/1,6 has separate effects, especially on innate immunity.

It’s a coin toss on whether observed cognitive improvement was due to 7% β-glucan soluble fiber, 7% indigestible fiber, or both since they were part of the same HFBG diet. I eat both fibers, beginning with Avena nuda oats for breakfast.

Astaxanthin bioavailability

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

By request, research on astaxanthin bioavailability. I used a “astaxanthin” “bioavailability” “quinone reductase” 2021 search term, and read citing papers.

“The bioaccessibility, bioavailability, and antioxidative activities of three astaxanthin geometric isomers were investigated using an in vitro digestion model.

  • 13Z-Astaxanthin showed higher bioaccessibility than 9Z- and all-E-astaxanthins during in vitro digestion, and
  • 9Z-astaxanthin exhibited higher transport efficiency than all-E- and 13Z-astaxanthins.

These might explain why 13Z- and 9Z-astaxanthins are found at higher concentrations in human plasma than all-E-astaxanthin.

9Z- and 13Z- astaxanthins exhibited a higher protective effect than all-E-astaxanthin against oxidative stress.”

https://pubs.acs.org/doi/10.1021/acs.jafc.7b04254 “Bioaccessibility, Cellular Uptake, and Transport of Astaxanthin Isomers and their Antioxidative Effects in Human Intestinal Epithelial Caco-2 Cells” (2017, not freely available)

jf1c00087_0005

“Astaxanthin with a high proportion of Z-isomer (especially rich in 9Z- and 13Z-isomers) was prepared from (all-E)-astaxanthin by thermal treatment and solid–liquid separation. Z-isomer-rich astaxanthin diet resulted in higher levels of astaxanthin in blood and many tissues (in particular, skin, lung, prostate, and eye) compared to all-E-isomer-rich diet.

Z-isomer-rich diet enhanced the level of 13Z-isomer in blood and tissues rather than that of 9Z-isomer. (13Z)-astaxanthin would have higher bioavailability and tissue accumulation than other isomers.”

https://pubs.acs.org/doi/10.1021/acs.jafc.1c00087 “Z-Isomers of Astaxanthin Exhibit Greater Bioavailability and Tissue Accumulation Efficiency than the All-E-Isomer” (2021, not freely available)

“Astaxanthin is highly susceptible to light, oxygen, and heat stress degradation. In addition, poor water solubility and bioavailability limit its efficacy in vivo. Investigating novel astaxanthin delivery systems is necessary in order to solve these drawbacks.”

https://www.mdpi.com/1420-3049/24/14/2640/htm “The Neuroprotective Effects of Astaxanthin: Therapeutic Targets and Clinical Perspective” (2019)

“Astaxanthin Z-isomers potentially have greater bioavailability and biological activity than (all-E)-astaxanthin. However, stability of Z-isomers is lower than all-E-isomer, which is a serious problem affecting its practical use.

In this study, we investigated impacts of different suspension media (oils and fats) and additives on astaxanthin isomer stability.

  • Z-isomers of astaxanthin isomerized to all-E-isomer during storage.
  • When soybean and sunflower oils were used as the suspension medium, astaxanthin isomers were hardly degraded. However the total Z-isomer ratio decreased from ~80% to ~50% during 6-week storage at 30 °C.
  • (9Z)-astaxanthin showed higher stability than 13Z- and 15Z-isomers.”

https://www.sciencedirect.com/science/article/abs/pii/S0308814621003770 “Evaluation and improvement of storage stability of astaxanthin isomers in oils and fats” (2021, not freely available)

I looked for but didn’t find a graph similar to this one that comparatively plotted astaxanthin:

OMCL2019-2716870.006

I also didn’t find recent human studies.

It seems that a special delivery system is required for taking astaxanthin as a supplement. It would require investigating manufacturers’ claims about isomer content and stability.

Eating colorful seafood is another way to get astaxanthin. Don’t know about eating raw or dried algae.

Ride the waves of gene expression with betaine

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

This 2021 cell study investigated a dietary supplement’s role in preventing nerve disease:

“A loss of epigenetic control has been implicated in development of neurodegenerative diseases. Previous studies have implicated aberrant DNA and histone methylation in multiple sclerosis (MS) disease pathogenesis.

We have previously reported that methyl donor betaine is depleted in MS and is linked to changes in histone H3 trimethylation (H3K4me3) in neurons. We have also shown that betaine increases histone methyltransferase activity by activating chromatin bound betaine homocysteine S-methyltransferase (BHMT).

A hallmark of MS is the death of oligodendrocytes, the cells responsible for wrapping axons in myelin in the central nervous system and maintaining a healthy sheath. In demyelinating diseases like MS, oligodendrocyte progenitor cells (OPCs) fail to differentiate and make more myelin, resulting in sclerotic lesions.

Promoting differentiation of OPCs and generation of myelin is of great interest as a novel MS therapy. Waves of gene regulation (repression and activation) need to occur to promote myelination.

This BHMT-betaine methylation pathway ensures availability of S-adenosylmethionine (SAM) for a variety of DNA and histone methylation processes. OPC survival and differentiation are dependent upon DNA and histone methylation, and both processes require SAM.

journal.pone.0250486.g001

BHMT uses betaine to remethylate homocysteine to methionine. Betaine can be taken in through the diet or synthesized through the oxidation of choline in mitochondria.

We demonstrated that oligodendrocyte gene expression can be modulated by betaine supplementation through the BHMT-betaine methylation pathway. Our study suggests that dietary betaine supplementation may prove to be a therapeutic agent for MS and other demyelinating disorders.”

https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0250486 “The BHMT-betaine methylation pathway epigenetically modulates oligodendrocyte maturation”

I started taking betaine 16 years ago. Didn’t know of these effects until reading this study.

Treating psychopathological symptoms will somehow resolve causes? had more on betaine (aka trimethyl glycine). Current dose is 1.5 grams twice daily.

Are rodent models of human neurodegenerative diseases realistic?

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

This 2020 stem cell review argued against rodent models of human neurodegenerative diseases:

“Neuronal loss is not caused solely by intrinsic degenerative processes but rather via impaired interactions with surrounding glia and other brain cells. Dysfunctional astrocytes do not provide sufficient nutrients and antioxidants to neurons, while dysfunctional microglia cannot efficiently clear pathogens and cell debris from extracellular space, resulting in chronic inflammatory processes in the brain.

Human glia, especially astrocytes, differ significantly in morphology and function from their mouse counterparts. Recent advances in stem cell technology make it possible to reprogram human patients’ somatic cells to induced pluripotent stem cells (iPSC) and differentiate them further into patient‐specific glia and neurons, thus providing a source of human brain cells.

stem3309-fig-0002-m

Astrocytes do not efficiently utilize energy resources and cannot provide adequate metabolic support to neurons. A coculture of healthy human neurons with diseased astrocytes impaired neuronal calcium responses to glutamate and γ‐aminobutyric acid (GABA) as compared to coculture with healthy human astrocytes.

Treatment with sulforaphane:

  • Normalized basal level glycolysis;
  • Decreased basal level Aβ42 secretion; as well as
  • Ameliorated inflammatory response to pro‐inflammatory cytokines TNF-α and IL1-β in PSEN1 mutant iPSC astrocytes.

It is essential to make sure that what we see in the dish is the real patient‐specific phenotype. Transplantation of human brain organoids containing microglia into mice could provide a novel tool for drug screening in vivo.”

https://stemcellsjournals.onlinelibrary.wiley.com/doi/full/10.1002/stem.3309 “Metabolic and immune dysfunction of glia in neurodegenerative disorders: Focus on iPSC models”

This review’s thesis seems plausible. However, one problem with in vitro stem cell studies is that they often don’t have a control group.

Giving children allergies with pets

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

This 2021 human study investigated development and persistence of allergies:

“Allergic rhinitis (AR) is a common IgE-mediated disorder involving troublesome symptoms of nasal congestion, nasal itch, sneezing, and associated eye symptoms. Like many chronic health conditions, AR stems from complex gene–environment interactions.

130 subjects with AR were recruited. Control population included 154 healthy children who underwent a regular physical examination in the same ear, nose and throat clinic as AR patients. Individuals with history of asthma or atopic dermatitis were excluded.

AR analysis

Plenty of contradictory associations exist as whether furred pet exposure (cats and dogs) may be a risk or a protective factor for AR development. Discrepancies are likely due to the ubiquitous nature of pet allergens, while pet owners are more concerned about sanitation and many other hygiene-related reasons.

Interaction of early-life pet exposure with methylation level of ADAM33 increased the risk for AR onset 1.423 times more in children. This study provides evidence that:

  • Early-life pet exposure and low methylation level of ADAM33 increase AR risk in children; and
  • The interaction between pet exposure and methylation level of ADAM33 may play an important role in development of AR.”

https://aacijournal.biomedcentral.com/articles/10.1186/s13223-021-00526-5 “Interaction between early-life pet exposure and methylation pattern of ADAM33 on allergic rhinitis among children aged 3–6 years in China”

There’s nothing children can do about who their parents were. Exposing them to pet allergens, though, may be another example of early-life experiences causing lifelong effects.

Happy Mothers Day

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

This 2021 rodent study investigated effects on offspring of maternal high-fat diet (HFD) during gestation and lactation, and offspring HFD during young adulthood:

“We found that gestation was the most sensitive period to induce obesity in late life, and there was no difference between sexes in chance of obesity. Furthermore, we found that lactation and administration of a HFD post‐weaning increased incidence of lipid metabolism disorders and obesity in offspring.

gestational hfd effects on offspring

There are different windows of opportunity for programming epigenetically labile genes. Some studies support the alteration of epigenetic status during development as an important cause induced adult obesity.

Gestation is considered as the most sensitive period because high DNA synthesis and DNA methylation patterns are established for normal tissue development during the embryonic period. These two programming events are the times when the epigenetic state changes most widely in the life cycle.”

https://onlinelibrary.wiley.com/doi/10.1111/jcmm.16551 “Gestational high-fat diet impaired demethylation of Pparα and induced obesity of offspring”

Hey mothers! Do what you please. But don’t turn around and deny consequences of your behavior and choices on your descendants’ physiology and behavior, and possibly those of further descendants.

Gestation, birth, infancy, and early childhood are critical periods for humans. There’s no going back to correct errors and problems.

Does skin improvement cause overall effects?

gettingwell4 4-5 stars Advanced knowledge, Epigenetics, Stress

This 2019 human skin study found:

“We demonstrated in aged mice that epidermal dysfunction largely accounted for age-associated elevations in circulating cytokine levels, and that improving epidermal function reduced circulating cytokine levels. We performed a pilot study to determine whether improving epidermal function reduces circulating proinflammatory cytokine levels in aged humans.

Both aged human and mouse skin display sustained abnormalities in epidermal permeability barrier homeostasis, stratum corneum (SC) hydration, and elevations in SC pH 4-6, each of which has been shown to independently provoke cutaneous inflammation. Disruption of the epidermal permeability barrier provoked an increase in:

  1. Cutaneous cytokine production; and
  2. Serum cytokine levels, independent of hepatic or T cell involvement.

We assessed whether improving epidermal function with an emollient, containing a mixture of lipids that mimics components of normal SC, lowered circulating levels of these same pro-inflammatory cytokines in aged humans.

skin treatment

After 30 days of twice-daily topical treatments, circulating levels of IL-1β and IL-6 decreased significantly in the treated aged cohort vs. untreated aged controls. Topical treatments reduced circulating levels of IL-1β and IL-6 to levels comparable to young controls. Though levels of TNF-α declined by over 40% in comparison to untreated aged humans, the difference did not attain statistical significance.

Results of this preliminary study suggest that a larger clinical trial should be performed to confirm whether improving epidermal function also can reduce circulating proinflammatory cytokine levels in aged humans, while also possibly attenuating downstream development of chronic inflammatory disorders.”

https://onlinelibrary.wiley.com/doi/abs/10.1111/jdv.15540 “Topical applications of an emollient reduce circulating pro‐inflammatory cytokine levels in chronically aged humans: a pilot clinical study” (not freely available)

I discussed enrolling in a trial whose objective would be to test this study’s findings. No big deal, just have to take IL-6 and TNF-α measurements in an upcoming annual physical. Then apply that trial’s skin treatment for 30 days per this study’s twice-daily protocol.

Day 70 results from Changing to a youthful phenotype with broccoli sprouts provided some of last year’s measurements. IL-6 was already at a negligible 1.0 pg / ml, one-fifth of both the above Baseline Young group’s 5.1 ± 0.9 and the Treated Aged Group’s 5.7 ± 0.9.

IL-6 2020

Probably won’t want my data, since their treatment wouldn’t be expected to lower an already very low inflammation marker.

Weight loss for the lazy

gettingwell4 4-5 stars Advanced knowledge, Epigenetics, Stress

At the risk of becoming Dr. Paul Clayton’s echo chamber, another great blog post, Falling Down:

“When lab rats or mice are weighted down with lead pellets they lose substantial amounts of weight, almost exclusively adipose tissue. Unlike dieting, there is little if any loss of muscle mass, making lead an ideal weight loss strategy for the lazy.

A clinical trial generated the same result. Their paper concludes, ‘Increased weight loading reduces body weight and fat mass in obese subjects in a similar way as previously shown in obese rodents. These findings demonstrate that there is a loading-dependent homeostatic regulation of body weight, the gravitostat, also in humans.’

Polyphenol resveratrol protects against damaging effects of de-loading by acting as an exercise mimetic, and does so by activating AMP-K directly. Other nutrients which do the same thing include polyphenol quercetin, sapogenin dammaranes, and omega 3 fatty acid EPA.”

The doctor still doesn’t mention sulforaphane in this or any other blog post, although it activates the AMPK pathway on the way to its primary effect of Nrf2 activation. First time I’d seen the term covidiots.

“You can fool some people sometimes
But you can’t fool all the people all the time
And now you’ve seen the light
Stand up for your rights”

Part 2 of Broccoli sprouts activate the AMPK pathway

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

This 2021 review subject was metformin’s role in autophagy:

“Metformin had been used as the first choice for treating diabetes for almost a century. Autophagy is responsible for recycling and degrading cellular components, which significantly affects cell functions in physiology and pathology.

Effects of metformin on autophagy mainly depend on corresponding signaling pathways in specific organs or tissues. Metformin can induce autophagy in cells of many organs and tissues via affirmed signaling pathways, such as AMPK-related signaling pathways.

1-s2.0-S0753332221000718-gr5_lrg

Different signaling pathways (alone or in combination) mediated the process of metformin affecting autophagy in different organs or tissues. It is necessary to combine effects of metformin on autophagy with pharmacological effects on pathologies in different organs or tissues, which would provide indications for future metformin applications.”

https://www.sciencedirect.com/science/article/pii/S0753332221000718 “The effects of metformin on autophagy”

I characterized this review as Part 2 of Broccoli sprouts activate the AMPK pathway because that study’s experimental evidence showed sulforaphane activation of the AMPK pathway was a predecessor to sulforaphane’s main effects of Nrf2 pathway activation. This review didn’t even mention Nrf2 activation.

Do all of metformin’s cited effects apply to daily intake of broccoli sprouts? Probably not, but most people who take metformin every day aren’t healthy.

See Part 3 for updates.

Grow your 3-day-old sprouts in darkness

gettingwell4 4-5 stars Advanced knowledge, Epigenetics

This 2021 study examined light frequency effects on Chinese kale sprouts’ development of glucosinolates:

“We investigated sprout growth and secondary metabolite glucosinolates (GSs) accumulation under white or combined red-and-blue (RB) light sources. Most GSs in sprouts are stored in seeds, which is gradually degraded to provide nutrients for other metabolic functions.

Phenotype of 3-day-old Chinese kale sprouts grown with different photoperiods condition under white or RB light:

capital A was grown in darkness

Sprouts grown under dark conditions showed only elongation of hypocotyls [shoots]. Sprouts grew with shorter hypocotyls and wider cotyledons [first leaves] irrespective of whether a white or combined RB light source was used.

Growth indicators (including plant height, cotyledon length, fresh weight, and dry weight) under different photoperiodic treatments were measured on days 2, 3, 6, and 9. Consistent with the phenotype presented, plant height and cotyledon length responded rhythmically to illumination time.”

https://www.frontiersin.org/articles/10.3389/fpls.2020.589746/full “Effect of Photoperiod on Chinese Kale (Brassica alboglabra) Sprouts Under White or Combined Red and Blue Light”

Circadian rhythms rule. Accept and adjust.

Week 56 of Changing to a youthful phenotype with sprouts

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

1. Per Improving healthy compounds of broccoli sprouts and Broccoli sprouts’ immune effects, this week I added mustard sprouts and red cabbage sprouts to my twice-daily routine of eating 3-day-old microwaved broccoli sprouts.

At first, I started mustard and red cabbage seeds with the same 10.7 gram weight (one tablespoon) of seeds. They grew well such that after three days, mustard sprouts weighed an average 61.2 g, and red cabbage sprouts weighed 60.3 g average. Both of these were slightly less than broccoli sprouts’ 65.5 g average.

3-day-old mustard sprouts substantially mellowed out from mustard seeds’ effects. After microwaving mustard sprouts to ≤ 60°C (140°F) and letting them sit for five minutes, I still felt constant nose burn while eating them. 3-day-old red cabbage sprouts were milder than broccoli sprouts, so no difficulties.

The main problem with doing one tablespoon seed weights of all three Brassicaceae species consistently was that 61.2 + 60.3 + 65.5 = 187 g (6.6 ounces) twice a day was too much for me. I eat a lot of low-calorie fibrous food everyday to make my gut microbiota happy. An extra 4+ oz increase at the same time as twice-daily broccoli sprouts put my stomach over the top.

I changed to make equal contents (one teaspoon) of these three Brassicaceae species be the 10.7 g (one tablespoon) that I started sprouting twice a day.

2. I haven’t seen relevant mustard and red cabbage 3-day-old sprout studies, only 7+ day microgreen and mature plant studies. Evidence is limited in determining effects of cutting my estimated 52 mg of daily sulforaphane intake from broccoli sprouts by two-thirds starting this week.

A. I’ve eaten a clinically-relevant amount of sulforaphane every day for 4+ times longer than any clinical trial. I’ve experienced many positive effects described in studies, and look forward to further improvements.

Reducing sulforaphane intake from broccoli sprouts to 17 mg is still within boundaries of measurable effects. As an example, Upgrade your brain’s switchboard with broccoli sprouts found effects from a daily sulforaphane 17.3 mg (100 µmol) intake. Plus red cabbage’s main glucosinolate, like broccoli sprouts, is glucoraphanin, which may hydrolyze to sulforaphane.

B. Mustard’s main glucosinolate, sinigrin, hydrolyzes to allyl isothiocyanate, and is in the same aliphatic group as broccoli’s glucoraphanin, which hydrolyzes to sulforaphane. An example of their similar effects was in a citation of Eat broccoli sprouts for DIM:

“Isothiocyanates are both inducers and substrates for Phase II enzymes as glutathione-S-transferases, and polymorphisms of these enzymes have a significant impact.”

Mustard’s myrosinase enzyme activities over and above broccoli myrosinase were highlighted in cited studies of Does sulforaphane reach the colon? Don’t know whether mustard sprouts’ myrosinase ≤ 60°C boosts broccoli and red cabbage sprouts’ hydrolyzation of glucoraphanin into sulforaphane.

C. Here’s a graphic from a 2010 study RED CABBAGE, A VEGETABLE RICH IN HEALTH-RELATED GLUCOSINOLATES which compared red cabbage glucoraphanin content with white cabbage:

red cabbage glucoraphanin vs white cabbage

The seeds I received were an “Agnostic” variety. In clarification correspondence with my supplier, I received a response “It means in this use ‘Generic’ or Variety not stated. Meaning it is just whatever variety of Red cabbage we bought and we don’t know the exact specifics.” 🙄

Red cabbage anthocyanins are greater than broccoli anthocyanins, which was highlighted in Colorize your diet, Red cabbage pigments and the brain, and Measuring bioavailability.

Figure 5 of Lab analyses of broccoli sprout compounds had analysis of three red cabbage cultivars’ 9-day-old sprouts. Glucosinolates are on top, hydrolysis products on the bottom. Glucoraphanin is red 4MSOB in A, and sulforaphane is red 4MSOB-ITC in C:

red cabbage 9-day-old sprouts

D. In summary, I don’t think I’ve significantly reduced broccoli sprouts’ effects by substituting two-thirds weight with two other Brassicaceae species. I haven’t noticed that growth characteristics / compounds interfered with each other.

Still looking for mustard and red cabbage 3-day-old sprout studies. My current Brassicaceae species composite is tasty, and doesn’t cause mustard nose burn.

3. This Brassicaceae species composite isn’t photogenic:

PXL_20210502_214348538

Red cabbage sprouts by themselves are pretty.

PXL_20210504_212505224

4. I still eat 3-day-old oat sprouts twice a day per Sprouting hulled oats. I don’t eat them with Brassicaceae species, but with Avena nuda oats in the morning, and AGE-less chicken vegetable soup in the evening.

One aspect of research on short-chain fatty acids

gettingwell4 2-3 stars Required further work, Cortisol, Dopamine, Epigenetics, Hippocampus, Hypothalamus, Immune system, Limbic system, Memory, Neurochemicals, Stress , ,

To further understand An overlooked gut microbiota product, a 2018 rodent study found:

“Microbial metabolites short-chain fatty acids (SCFAs) have been implicated in gastrointestinal functional, neuroimmune regulation, and host metabolism, but their role in stress-induced behavioural and physiological alterations is poorly understood

SCFAs are primarily derived from fermentation of dietary fibres, and play a pivotal role in host gut, metabolic and immune function. All these factors have previously been demonstrated to be adversely affected by stress.

Administration of SCFAs to mice undergoing psychosocial stress alleviated enduring alterations in anhedonia and heightened stress-responsiveness, as well as stress-induced increases in intestinal permeability.

experimental design

SCFA treatment alleviated psychosocial stress-induced alterations in reward-seeking behaviour, and increased responsiveness to an acute stressor and in vivo intestinal permeability. In addition, SCFAs exhibited behavioural test-specific antidepressant and anxiolytic effects, which were not present when mice had also undergone psychosocial stress.”

https://physoc.onlinelibrary.wiley.com/doi/pdf/10.1113/JP276431 “Short-chain fatty acids: microbial metabolites that alleviate stress-induced brain–gut axis alterations”

One way researchers advance science is to relate aspects of their findings to previous studies. That approach works, but may miss items that weren’t covered in previous research.

This study fed specific quantities of three SCFAs – acetate, butyrate, and propionate – apparently due to previous research findings. If other SCFAs produced by gut microbiota were ignored – like crotonate (aka unsaturated butyrate) – how would that approach advance science?

I found this study from its citation in Harnessing endogenous defenses with broccoli sprouts.