Increase broccoli compound amounts with thermosonication

This 2021 study combined ultrasound and heat to increase broccoli compounds:

“A large proportion of broccoli biomass is lost during primary production, distribution, processing, and consumption. This study evaluated thermosonication for pre-treatment of broccoli florets to enhance enzymatic conversion of glucoraphanin (GR) into bioactive sulforaphane (SF).

Our results showed that a combination of ultrasound with heat treatment (at 50 and 60 °C for 5 and 7 min) improved release of GR from broccoli matrix over and above heat treatment alone. This could be due to mechanical disruption of plant cell wall and plant matrix, which enhances release of intracellular content.

The highest residual level of both GR (126.3% increase) and sulforaphane was observed after thermosonication at 60 °C for 7 min. It can be concluded that:

  1. Measured levels of glucoraphanin in untreated broccoli using current methods gives only an indication of the amount of easily accessible and extractable glucoraphanin in broccoli; and
  2. Processing of broccoli by heat and ultrasound enables release of possibly bound glucoraphanin and enhances conversion to sulforaphane.

Higher sulforaphane yield was observed under conditions of significant myrosinase inactivation. This could be attributed to:

  • Inactivation of ESP [epithiospecifier protein], providing a suitable condition for synthesis from GR to SF as opposed to SF-nitrile; and/or
  • Increased in situ residual myrosinase activity at elevated temperatures; and/or
  • Increased release and accessibility of GR.

Maximum SF content among studied conditions was achieved using thermosonication treatment at 60 °C for 7 min, which represented a 192.6% increase compared to untreated broccoli florets. That was 15.8% higher than thermal treatment, indicating the potential of thermosonication for enhancing sulforaphane yield over and above effects of heat.”

https://www.mdpi.com/2218-273X/11/2/321/htm “Thermosonication for the Production of Sulforaphane Rich Broccoli Ingredients”


I mentioned in Week 34 how broccoli sprout experts tried to smear a Microwave broccoli to increase sulforaphane levels finding of more glucoraphanin with increasing temperatures after microwaving. This study measured a similar effect of glucoraphanin increases using two techniques:

“For GR extraction two methods were initially examined: aqueous extraction or methanolic extraction. We wanted to compare effects of using different extraction solvents to evaluate whether levels of extractable GR were influenced by the type of solvent.

The difference between GR content of aqueous and methanolic extracts was not statistically significant.”

😉

Thermosonication isn’t suitable for home use. Microwaving is.

A microwave can’t keep contents at a constant 60° C for seven minutes, though. Optimizing myrosinase activity is also a consideration for increasing broccoli sprout compounds at home.

Microwave broccoli sprouts to increase sulforaphane

This 2020 review explored sulforaphane stability and formation:

“Sulforaphane (SF) is beneficial to our health since it can reduce incidence of a number of tumors, induce cell cycle arrest and apoptosis in multiple experimental models. However, since neither SF nor myrosinase is thermostable, it is essential to increase stability of SF and/or enhance conversion of glucoraphanin (GRP) to SF by myrosinase to maximize SF therapeutic benefits.

  • Since little or no water is needed for microwaving and stir-frying, and broccoli is not immersed in water during steaming, SF content in broccoli florets is higher in these processes compared to that in boiled florets.
  • Thermostability of Brassicaceae myrosinase varies across different species and cultivars, as well as the plant organ. For example, myrosinase in broccoli florets are more thermosensitive compared to that in sprouts, likely due to the presence of a seed-specific myrosinase.
  • GRP in cooked broccoli can be hydrolyzed by intestinal microbiota to SF, sulforaphane nitrile, and/or other isothiocyanates and nitriles, although the decomposition rate is very low. However, continuous feeding of rats or mice with broccoli increased myrosinase-like activities in colon and cecum contents.

Mild heating (40-60 °C) by microwave increased SF content in broccoli. High-power microwave heating with temperature control at 60 °C could retain higher bioavailability.

Continuous broccoli ingestion enhances myrosinase-like activity of gut microbiota. Regardless of differences in endogenous amounts of glucosinolates and myrosinase across multiple broccoli varieties, moderate microwaving ensures optimum SF availability.”

https://www.sciencedirect.com/science/article/abs/pii/S0308814620326339 “Approaches for enhancing the stability and formation of sulforaphane” (not freely available)


Several studies previously curated were referenced, including:

  1. Microwave broccoli seeds to create sulforaphane;
  2. Microwave broccoli to increase sulforaphane levels;
  3. Enhancing sulforaphane content.

Why wait around for approval or citation or social validation of research? When Study 2 showed microwaving increased sulforaphane and glucoraphanin content this time last year, I followed the evidence and acted accordingly.

Researchers who deny microwaving’s beneficial effects on broccoli compounds up to 60° C can publish contrary findings, or keep their biased opinions to themselves. See Increase broccoli compound amounts with thermosonication for further evidence.

PXL_20210327_113713116

Year One of Changing to a youthful phenotype with broccoli sprouts

1. My subjective experiences these past 52 weeks are that noticeable changes keep happening due to combinations of:

  • Eating clinically-relevant, twice-daily doses of microwaved 3-day-old broccoli sprouts, and taking nothing else an hour before or an hour after;
  • Eating 3-day-old oat sprouts twice a day;
  • Exercising every day;
  • Taking yeast cell wall β-glucan and other supplements twice a day;
  • Eating AGE-less chicken vegetable soup twice a day;
  • Eating 81+ grams (dry weight before soaking 12+ hours) of Avena nuda oats for breakfast;
  • Working from home 40 hours a week.

I look forward to more evidence on youthening during Year Two.

2. The three previous blog posts concerned yeast cell wall β-glucan. I haven’t received expected results from my New Year’s resolution to be prepared for Spring’s allergy onslaught. Actions included:

  • Changing diet to be more gut-friendly;
  • Taking WellImmune β-glucan in a morning dose of 500 mg for over two months;
  • Continuing with another company’s β-glucan in an evening 400 mg dose.

Consider this graphic from a third competitor regarding their Glucan 300 β-glucan:

dosage

My weight is probably 150-ish. I’d take three or four 500 mg capsules daily if I used Glucan 300. β-glucan comparisons evaluated it as the most active of five dozen commercially available products.

My lack of expected 2021 allergy results may be due to both insufficient dose and less-active products. Glucan 300 discloses its active content in specific percentages of 1,3/1,6 terminal-linked glucose molecules:

  1. Yeast cell wall β-glucan’s effects depend on biologically active content.
  2. Companies with evidence-based competitive products will disclose these contents.
  3. Neither β-glucan product I currently take does that.

I’ll use Glucan 300 exclusively when I receive it later this week. Can’t grow a fire by piling on more wet green wood if I’m serious about improving my innate and adaptive immune systems. Allergy season continues another few weeks, so it shouldn’t take long to find out if this change makes a difference.

PXL_20210327_110807824


Evaluating a company-sponsored β-glucan paper

This 2020 review subject was yeast cell wall β-glucan effects in humans:

“The first aim of this review is to collate and interpret the existing pre‐clinical research on β‐1,3/1,6‐glucan with regard to immunity in order to clarify its molecular mechanism of immunomodulatory action.

mnfr3715-fig-0001-m

The second aim of this review is to collate and evaluate the literature in order to provide a comprehensive overview of human studies assessing the effect of supplementation with high quality, well‐characterized β‐1,3/1,6‐glucan from commercially available sources on immunity across multiple populations. Inclusion criteria consist of randomized, double‐blind, placebo‐controlled human studies that investigated efficacy of orally administered β‐glucan with a purity of over 75%.”

https://onlinelibrary.wiley.com/doi/10.1002/mnfr.201901071 “β‐1,3/1,6‐Glucans and Immunity: State of the Art and Future Directions”


I don’t usually curate company-sponsored research, aka puff pieces. I wondered why, after taking WellImmune β-glucan 500 mg daily for over two months, I didn’t have expected results.

There are always several possible explanations for experimental failures. I didn’t see applicable items in this paper.

There was much information regarding things their sponsor’s customers don’t need to know. Just like their sponsor’s product label, there was little about what customers need to know, such as:

What was each product’s content, in specific percentages, of 1,3/1,6 terminal-linked glucose molecules? That makes a difference.

The sponsor knows, but doesn’t disclose it on their product’s label. These researchers could have found out and presented that information on their sponsor’s and other companies’ products for each study reviewed.

Not doing so deprived readers of an important evaluation criteria that could possibly explain variable results and provide a better measure for comparability. Stopping at “a purity of over 75%” instead of investigating and disclosing exact information was evasive.

Choosing your future with β-glucan

This 2020 rodent study investigated yeast cell wall β-glucan effects on bacterial infections:

“β-glucan is a potent inducer of epigenetic and functional reprogramming of innate immune cells, a process called trained immunity, resulting in an enhanced host response against secondary infections. We investigate whether β-glucan exposure confers protection against pulmonary Mycobacterium tuberculosis (Mtb) infection.

  • β-glucan induces trained immunity via histone modifications. β-glucan-induced trained immunity confers protection against virulent Mtb via the IL-1 signaling pathway.
  • β-glucan-induced trained immunity enhances production of proinflammatory cytokines in human monocytes challenged with heat-killed Mtb. Increase in cytokine production capacity was the result of epigenetic reprogramming and mediated via the PI3K/Akt/mTOR pathway.

Most important, β-glucan-treated mice infected with Mtb demonstrated remarkably enhanced survival, which was dependent on IL-1 signaling.

survival Mtb

β-glucan epigenetically reprograms human monocytes, leading to a phenotype characterized by a unique IL-1 signature and anti-mycobacterial properties. β-glucan-treated mice were protected against pulmonary Mtb infection.

While both β-glucan and BCG [Bacillus Calmette-Guerin tuberculosis vaccine] reprogram HSCs to induce trained immunity, BCG reprogramming of HSCs was dependent on IFNγ signaling. β-glucan reprogramming of HSCs was mediated via IL-1 signaling, which was also required for protection against Mtb infection.

Considering safety of β-glucan in a human clinical trial, our results strongly suggest potential clinical implications of β-glucan for both prophylactic and therapeutic use in TB.”

https://www.cell.com/cell-reports/fulltext/S2211-1247(20)30587-8 “β-Glucan Induces Protective Trained Immunity against Mycobacterium tuberculosis Infection: A Key Role for IL-1″


My comment “many of these findings also apply to yeast cell wall β-glucan treatments” in Long-lasting benefits of a common vaccine lacked clarity. This post provides part of that evidence.

So where do you choose to be? In an 80% survival group who were administered β-glucan before they encountered a serious infection? Or in a < 20% survival group who didn’t take β-glucan?

Which is better for resolving a health situation before it becomes a problem?

  • Roll the dice, and hope for luck / providence?
  • Do nothing constructive, and depend on interventions after a problem occurs?
  • Take responsibility for your own one precious life?

PXL_20210327_111214712

β-glucan comparisons

This 2018 rodent study compared and contrasted commercially available β-glucans:

“β-glucans are natural biologically-active compounds called ‘biological response modifiers.’ This study is a follow-up of our three previous studies that tested 43 different glucans.

We used 16 different glucans isolated from yeasts, mushrooms, algae, and oats. We compared their effects on phagocytosis, IL-2 production, antibody secretion, and inhibition of three experimental cancer models.

16 beta glucans

Our results showed significant differences among tested glucans, despite the fact that glucans in general have strong stimulating effects on most aspects of the immune system. Differences between activities of commercially available glucans might be an explanation for the sometimes confusing results found in the literature. In all tests employed, Glucan #300 was the most active.

Highly purified and active glucans have significant pleiotropic effects.”

https://www.researchgate.net/publication/323523231_Glucans_and_Cancer_Comparison_of_Commercially_Available_b-glucans_-_Part_IV “Glucans and Cancer: Comparison of Commercially Available β-glucans – Part IV” (registration required)


PXL_20210327_103105655.NIGHT

Oat digestibility

A reader questioned one part of Oat species comparisons of the good stuff regarding Avena nuda hull digestibility. This 2019 study partially investigated that aspect:

“We investigated effects of proteins, lipids, and β-glucan in naked oat flour on in vitro digestibility of starch. Content of rapidly digested starch increased, and content of resistant starch decreased after removing non-starch constituents.

There are three categories of starch in accordance with the rate and degree of starch digestion, namely, rapidly digested starch, slowly digested starch, and resistant starch. Resistant starch cannot be digested. Instead, it promotes growth of beneficial colonic flora.

Digestibility of starch is influenced by size and shape of starch granules, food processing method, physical and chemical modifications, viscosity, and food matrix components. Physicochemical properties of naked oat starch and naked oat flour after removing non-starch constituents were compared to study relationships between starch digestibility and intrinsic factors:


Oats contain more proteins and lipids than other common grains. Proteins can effectively reduce starch digestibility by several mechanisms:

  • Proteins can form a protection around starch granules, restricting entry of enzymes into substrates.
  • Surface proteins can block catalytic binding of enzymes on starch granule exterior.
  • α-amylase can partially bind to proteins, reducing enzyme utilization.

By contrast, effects of lipids on starch digestibility is primarily due to forming complexes with amylose, which is better able to resist amylase.

β-glucan, particularly the extracted water-soluble fraction, can lower digestion rate of starch by increasing viscosity. β-glucan can create a complex of adjacent proteins to form a robust structure that resists amylase, resulting in a decrease in starch digestibility.”

https://www.sciencedirect.com/science/article/abs/pii/S0308814619310556 “Non-starch constituents influence the in vitro digestibility of naked oat (Avena nuda L.) starch” (not freely available)


When viewing the above graphic, keep in mind that its order wasn’t sequential. So “degreased” oat flour (lipids removed, DG-NOF) wasn’t included in “deproteinized” oat flour (DP-NOF).

This in vitro study missed an opportunity to investigate human-practical aspects. Nobody eats oats without preparing them with water. But effects on digestibility from minutes and hours of soaking, boiling, microwaving, etc. weren’t analyzed.

Gut microbiota outnumber human cells 10-to-1. Treat them well with both Avena nuda resistant oat starch and indigestible hulls, and expect reciprocity.

Oat species comparisons of the good stuff

This 2020 study compared and contrasted distributional compositions of two oat species’ seeds:

“Oat grains of one hulless variety (Lamont) with low avenanthramide (AVA) contents and one hulled variety (Reins) with high AVA contents were sequentially abraded. Contents of nutrients (protein, oil, starch, β-glucan, ash, and other carbohydrates) and AVAs were measured.

A relationship between content of a constituent in the surface layer abraded off (termed pearling fine, or PF) at each cycle of pearling and the cumulative level of surface removal could be established. This relationship essentially describes true distribution or localization of individual constituents across an oat kernel.

AVAs provide health benefits in mammals, including anti-oxidation, anti-inflammation, anti-atherosclerosis, and anti-cancer properties. Relationships between contents of four AVAs and total AVAs in pearling fines (A) and corresponding pearled kernels (B) of hulless Lamont oat [top] and hulled Rein [bottom] with cumulative surface removal levels achieved by sequential pearling:

For Lamont oat, AVAs 2c, 2f, 2p, 5p, and total AVAs all showed decreasing concentrations with increasing levels of surface removal. The first PF (4% surface removal) contained the highest amounts for all four AVAs, with 2p near ten times higher than in whole grain.

Hulled Reins oat differed significantly from hulless Lamont oat in not only amounts of AVAs but also their distribution patterns within kernels. Dehulling caused reduction in total AVA content.

Pearled oats contained less protein, oil, ash, and other carbohydrates and AVAs, but more starch than whole grain. In contrast, oat bran contained more AVAs, protein, oil, ash and other carbohydrates but less β-glucan and starch as compared to whole grain.”

https://www.sciencedirect.com/science/article/abs/pii/S0308814620315302 “Distributions of nutrients and avenanthramides within oat grain and effects on pearled kernel composition” (not freely available)


There were higher AVA contents in hulls of the top graphic’s species (Avena nuda) compared with its next ten seed layers. Humans require the bottom graphic’s oat species’ (Avena sativa) hull, which is “about 25% total grain mass,” to be milled off before we eat it. So AVA data points on the bottom graph A start around 25% surface removal.

As mentioned in Eat oats to prevent diabetes, I replaced steel-cut Avena sativa oats with whole Avena nuda oats for breakfast. I don’t know how well Avena nuda hulls are digested, but gut microbiota ferment similar indigestibles into beneficial compounds.

The first study of Eat oat sprouts for AVAs found “up to 25-fold increase” in AVAs with 7-day-old Avena sativa sprouts. I expect 3-day-old hulled Avena sativa sprouts I eat also increase AVAs as they germinate.

Long-lasting benefits of a common vaccine

This 2021 review subject was effects of the 100-year-old tuberculosis vaccine:

“Bacillus Calmette-Guerin (BCG) vaccine is one of the most widely used vaccines. It protects against many non-mycobacterial infections secondary to its nonspecific immune effects.

The mechanism for these effects includes modification of innate and adaptive immunity. BCG vaccine is known to not only boost immune responses to many vaccines when they are co-administered, but also decreases severity of these infections when used alone.

Alteration in innate immunity is through histone modifications and epigenetic reprogramming of monocytes to develop an inflammatory phenotype, a process called trained immunity. Memory T cells of adaptive immunity are also responsible for resistance against secondary infections after administration of BCG vaccine, a process called heterologous immunity.

The PI3K/AKT pathway, another pathway for mediating immunity, was upregulated. This was supported by recent studies demonstrating its involvement in induction of trained immunity by both BCG and β-glucan.

BCG vaccine can modify both innate and adaptive immunity, and provide immunity not only against Mycobacterium tuberculosis but also other pathogens. Heterologous immunity and trained immunity contribute to pathophysiologic mechanisms which explain how a vaccine protects against unrelated pathogens.”

https://www.amjmedsci.org/article/S0002-9629(21)00092-6/fulltext “Bacillus Calmette-Guerin Vaccine and Nonspecific Immunity”


As inferred by “induction of trained immunity by both BCG and β-glucan” many of these findings also apply to yeast cell wall β-glucan treatments. See Choosing your future with β-glucan for a representative study.

Our first 1000 days

This 2021 review subject was a measurable aspect of our early lives:

“The first 1000 days from conception are a sensitive period for human development programming. During this period, environmental exposures may result in long-lasting epigenetic imprints that contribute to future developmental trajectories.

The present review reports on effects of adverse and protective environmental conditions occurring on glucocorticoid receptor gene (NR3C1) regulation in humans. Thirty-four studies were included.

The hypothalamic-pituitary-adrenal (HPA) axis is key in regulating mobilization of energy. It is involved in stress reactivity and regulation, and it supports development of behavioral, cognitive, and socio-emotional domains.

The NR3C1 gene encodes for specific glucocorticoid receptors (GRs) in the mammalian brain, and it is epigenetically regulated by environmental exposures.

When mixed stressful conditions were not differentiated for their effects on NR3C1 methylation, no significant results were obtained, which speaks in favor of specificity of epigenetic vestiges of different adverse conditions. Specific maternal behaviors and caregiving actions – such as breastfeeding, sensitive and contingent interactive behavior, and gentle touch – consistently correlated with decreased NR3C1 methylation.

If the neuroendocrine system of a developing fetus and infant is particularly sensitive to environmental stimulations, this model may provide the epigenetic basis to inform promotion of family-centered prevention, treatment, and supportive interventions for at-risk conditions. A more ambiguous picture emerged for later effects of NR3C1 methylation on developmental outcomes during infancy and childhood, suggesting that future research should favor epigenome-wide approaches to long-term epigenetic programming in humans.”

https://www.sciencedirect.com/science/article/abs/pii/S0149763421001081 “Glucocorticoid receptor gene (NR3C1) methylation during the first thousand days: Environmental exposures and developmental outcomes” (not freely available). Thanks to Dr. Livio Provenci for providing a copy.


I respectfully disagree with recommendations for an EWAS approach during infancy and childhood. What happened to each of us wasn’t necessarily applicable to a group. Group statistics may make interesting research topics, but they won’t change anything for each individual.

Regarding treatment, our individual experiences and needs during our first 1000 days should be repeatedly sensed and felt in order to be therapeutic. Those memories are embedded in our needs because cognitive aspects of our brains weren’t developed then.

To become curative, we first sense and feel early needs and experiences. Later, we understand their contributions and continuations in our emotions, behavior, and thinking.

And then we can start to change who we were made into.

Week 50 of Changing to a youthful phenotype with broccoli sprouts

1. Effects of broccoli sprouts that seemed personally astounding at Week 10 became a part of day-to-day life. What will happen next?

Day 350 of eating a clinically relevant amount of broccoli sprouts every day seems like a large number. Yet in comparison, for 6,000+ days I’ve taken a clinically relevant dose of 1/3, 1/6 yeast cell wall β-glucan to train my innate immune system.

Both of their main actions are similar in mildly stimulating my body’s stronger defenses. Switch on your Nrf2 signaling pathway described sulforaphane’s effects as a “weak pro-oxidant signal that you use to activate Nrf2.” Take responsibility for your one precious life – β glucan described yeast cell wall β-glucan as “a potent immune response potentiator and modulator.”

2. I work a full-time job, in my 24th year of being paid to develop software. It’s a young-person’s field, contingent on learning new aspects of new languages, then performing every hour of every day up to what’s known or should be known.

I enjoy working with a group of talented individuals, especially when it involves creative problem solving. I’ll tolerate admin, limited meetings, and other things I don’t like when there’s a path toward doing what I enjoy.

My experiences since coming out of retirement to take this job four years ago have been analogous to Part 3 of Rejuvenation therapy and sulforaphane, i.e. old treated subjects learned and remembered significantly better than old untreated subjects, but not as well as untreated young subjects. I’m not a lab rat, though, and I’ve often had better performance since Week 10 than decades-younger coworkers.

All part of Surfacing Your Real Self.

3. Looking back on this week and month last year, there was worldwide herding of the population using a disease as a cover story. I wondered when it would end, and I’m still wondering because it’s still going on.

This pretext for surrendering human rights is easily derailed. I won’t enumerate fallacies, misrepresentations, frauds, assertions that lack evidence.

It’s a valuable skill for us to know when we’re being herded. Are you willing and able to develop that skill?

4. I didn’t disturb this heron, and was rewarded with a lightning-fast snatch-and-swallow of its breakfast. I wasn’t quick enough to get that photo, though.

Flailing with probiotics?

This 2021 review subject was probiotic bacteria survival and colonization:

“Health benefits of probiotics are diminished due to substantial reduction of viable probiotic bacteria under harsh conditions in the gastrointestinal tract and colonization resistance caused by commensal bacteria. This review illustrates the journey of probiotics from oral administration to the gastrointestinal tract, followed by colonization of the gut, with a particular focus on the adhesion process of probiotics on mucosa or intestinal epithelial cells.

  • Mouth – influence of saliva on survival rates of probiotics seems to be minimal.
  • Stomach – transit takes between 5 min and 2 h. Prolonged exposure to the acidic environment is a huge challenge for probiotics.

  • Small intestine – bile acids and digestive enzymes (including lipases, proteases, and amylases) can impact probiotic viability through cell membrane disruption and DNA damage.
  • Colon – probiotics compete with host microbiota for nutrients and adhesion sites to colonize colonic mucosa and proliferate. Due to colonization resistance, most probiotics are excreted so that they cannot be detected.

Composition of gut microbiota is highly variable. Microbial composition is considerably different between people in different geographic locations and with different diets.

Probiotics cannot change intestinal microbiota community structure or diversity.

How probiotics communicate with commensal bacteria and some are successfully introduced to gut microbiota is of great interest. Understanding these factors will facilitate employment of effective delivery strategies designed for probiotics to overcome colonization resistance and achieve health benefits.”

https://www.frontiersin.org/articles/10.3389/fcimb.2021.609722/full “Probiotic Gastrointestinal Transit and Colonization After Oral Administration: A Long Journey”


This review provided details supporting points 2 and 6 of Harnessing endogenous defenses with broccoli sprouts:

“Even though probiotics as food or supplements demonstrate favourable clinical outcomes, they typically don’t colonise the gut. How do we expect them to restore diversity and lost species to the gut microbiome after antibiotics? If no trace of an administered probiotic organism can be found a few weeks later, is there any sustained benefit?

If the gut can harbour around 1,000 different species, why do we expect a probiotic supplement harbouring just a few species to favourably modify a human microbiome?”

That paper’s emphasis was reflected in its title, “Restoring Gut Ecology: Harnessing the Inbuilt Defence Mechanisms of the Gut Epithelium.”


I stopped taking probiotics earlier this year after 16 years of twice-daily intake. I’ve increased prebiotic intake. Pretty soon I’ll find out whether my innate and adaptive immune systems have changed enough to ward off spring allergy-season effects.

Sand sculptures