Does reprogramming signaling pathways create memories?

This 2020 study investigated genes and signaling pathways for inflammatory memory:

“Fibroblast-like synoviocytes (FLS) play a critical role in pathogenesis of rheumatoid arthritis (RA). Chronic inflammation induces transcriptomic and epigenetic modifications that imparts a persistent catabolic phenotype to the FLS, despite their dissociation from the inflammatory environment.

Sustained activated genes established pro-inflammatory signaling components known to act at multiple levels of NF-κB, STAT and AP-1 signaling cascades. Sustained repressed genes included critical mediators and targets of the BMP [bone morphogenic protein] signaling pathway.

We identified sustained repression of BMP signaling as a unique constituent of the long-term inflammatory memory induced by chronic inflammation.

FLS are synovial tissue-resident and specialized mesenchymal cells critical for homeostasis. Key features of these cells during homeostasis include the production of extracellular matrix components and providing nutrients to the synovial fluid. Healthy synovium is composed of multiple layers of FLS, which forms the synovial lining and sublining through cell–cell contacts.

Inflammatory and pro-resolving mediators are tightly regulated to maintain normal synovium functioning. However, in inflammatory and autoimmune diseases such as rheumatoid arthritis, an imbalance between these signals causes homeostasis disruption leading to synovial tissue damage, cartilage destruction and bone degeneration.”

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7679373/ “Chronic exposure to TNF reprograms cell signaling pathways in fibroblast-like synoviocytes by establishing long-term inflammatory memory”


These researchers described a positive feedback loop that kept rheumatoid arthritis in place. No feedback diagram or explanation of what sustained a disease condition other than to say:

“Gene expression changes induced by short-term tumor necrosis factor-alpha (TNF-α) treatment were largely sustained in the FLS exposed to chronic inflammation.”

Okay – then what upstream signals sustained TNF-α? What would it take to interrupt that feedback loop? What initiated it?

Studies usually substantiate effects by also developing evidence for causes of opposite effects and of no effects. This study investigated neither reversibility nor no effect, and instead stated:

“Multiple signaling networks are irreversibly modified due to TNF-α-mediated long-term epigenetic and transcriptomic reprogramming. We speculate that sustained repression of BMP signaling may be critically required to ensure the persistently transformed phenotype of RA FLS.”

No evidence was offered for “irreversibly modified.” Anyway, that didn’t fit with:

“We postulate that simultaneous targeting of these activated and repressed signaling pathways may be necessary to combat RA persistence.”

Enduring epigenetic memories? Or continuous toxic stimulation? provided another perspective: “Enduring epigenetic effects may be symptoms rather than causes when toxic conditions persist.”


Been on a Steely Dan kick lately. Probably due to this year’s Royal Scam:

Zinc and broccoli sprouts – a winning combination

This 2019 study deserved better coverage than a one sentence mention in Reversal of aging and immunosenescent trends with sulforaphane:

“Obstructive sleep apnea syndrome is one of the most common breathing disorders in sleep, with a high prevalence of 3–7% and severe consequences. It is characterized by intermittent hypoxia (IH) due to recurrent episodes of partial or complete collapse of the upper airway during sleep, leading to blood hypoxemia, hypercapnia, sleep fragmentation, augmented respiratory efforts, and increased sympathetic activity.

Our study is the first investigation of the combination of BSE [broccoli sprout extract] and Zn [zinc], Nrf2, and MT [metallothionein] inducers, to protect against IH-induced cardiomyopathy. By effectively activating Nrf2, its downstream targets, and MT, this combination can ameliorate defects associated with IH-induced cardiomyopathy more effectively than monotherapies.

Mice were administered with BSE (equivalent to SFN [sulforaphane] 2 mg/kg) and/or Zn sulfate heptahydrate (5 mg/kg) by gavage from 8 weeks of age at a frequency of once every other day for 8 weeks. Doses used in this study are safe to convert to human doses. [2 mg x .081 x 70 kg = 11 mg sulforaphane; 5 mg x .081 x 70 kg = 28 mg zinc]

  • Heart mass was significantly lower in the IH-BSE/Zn group than in IH and IH-BSE groups. Heart mass / tibia length ratio was significantly lower in the IH-BSE/Zn group than in IH and monotherapy groups.
  • Treatment with BSE and/or Zn can ameliorate myocardial fibrosis associated with IH, to a certain extent, and combination therapy has the best antifibrotic effect among treatments.
  • BSE or Zn can significantly ameliorate myocardial inflammation induced by IH, but the combination provides a better anti-inflammatory effect.
  • We used 3-NT as an indicator of the severity of oxidative stress. 3-NT protein levels were significantly reduced in IH mice for all treatment groups, and reduction was greater in the combination treatment group.
  • Combination is more effective than monotherapies to activate Nrf2-mediated antioxidant function.

  • In Zn-treated and combination treatment groups, MT protein expression was significantly higher than in the IH group, and there was only a slight increase in the IH-BSE group.”

Combination of Broccoli Sprout Extract and Zinc Provides Better Protection Against Intermittent Hypoxia-Induced Cardiomyopathy Than Monotherapy in Mice


One way to improve broccoli sprout compounds’ effects is to eat them with zinc. One way to improve zinc’s actions is to take it with broccoli sprouts.

Week 34 of Changing to a youthful phenotype with broccoli sprouts

1. Thank you to readers of this blog who find the 650+ curations and other posts worth their time. I reread blog posts after you read them, and sometimes improve them for our mutual benefit.

One such post this week was Broccoli sprout compounds include sinapic acid derivatives. Although it was already fairly detailed, it received a half-dozen improvements.

  • Those researchers measured composition changes of 31 compounds (18 sinapic acid derivatives, 8 glucosinolates, and 5 flavonoids) identified in seed-2-4-6-day germination stages of one cultivar. They provided convertible dry weight and fresh weight measurements in mg / g.
  • It complemented the 3-day-old broccoli sprouts have the optimal yields study comparisons of six cultivated varieties’ seed-3-5-7-day germination stage weights and measurements with their origins using a milligram-per-gram-of-seeds scale:

    “To be comparable, the content of these bioactive compounds from 100 fresh sprouts was divided by the weight (g) of 100 seeds, and then this value was compared with their content from one gram seeds.”

  • The sinapic acid study discussed another study for:

    “In a study, diminishing amounts of total phenolic acids in sprouts of three broccoli cultivars was observed only between 3rd and 7th day of germination under photoperiod conditions and only when expressed on fresh weight basis. After recalculating results to dry weight, amounts were increasing during the whole 14-day observation period.”

All studies were scientifically informative. Still, results depended on researchers’ operative paradigms, and human behavior such as unconscious act-outs of unsatisfied needs to feel important.

2. Speaking of which, I viewed a 1:48 video with broccoli sprout experts who disparaged microwaving around the 1:10 mark. I’m not an expert, but I’ve eaten a clinically-relevant dose of microwaved broccoli sprouts every day for 34 weeks now.

Here are a few studies of microwaving’s effects on phenolic, glucosinolate, and flavonoid broccoli compounds. Just for those who value evidence more than opinion.

  • Microwaving broccoli sprouts may not affect phenolic levels found four of five test cases didn’t significantly diminish total phenolic fresh weight contents of whole broccoli. They blended 100 grams broccoli in 200 ml water, halved the purée, then microwaved half on 700W power for 30 seconds. No disclosure of what temperature was achieved, but it was probably < 60°C (140°F). Microwaving significantly increased the glucosinolate hydrolysis product indole-3-carbinol:

    “I3C in broccoli was increased by 3.1, 9.1 and 1.9 folds respectively using blenders 1, 2 and 5 with microwaving.”

  • Microwave broccoli to increase flavonoid levels study design was “Broccoli florets (150 g) were put in a microwave safe bowl with a 1 tablespoon [15 ml] of water” and a 1200W microwave on full power for one minute. Although this may have produced temperatures > 60°C, flavonoid fresh weight contents increased > 30%:

    Microwaving may increase extractability and/or release from binding to other compounds as a result of matrix softening.

  • Microwave broccoli to increase sulforaphane levels demonstrated significant differences for 450W (LL) and 900W (HL) power settings in glucoraphanin and sulforaphane dry weight amounts when broccoli florets were microwaved to the same temperatures. Compare white bar sulforaphane amounts for LL60 and HL60 (both 60°C), annotated as E and F:

    “Microwave treatment causes a sudden collapse of cell structure due to the increase in osmotic pressure difference over vacuole membrane. Microwave irradiation might help to release more conjugated forms of glucosinolates and then get hydrolyzed by released myrosinase.

  • Enhancing sulforaphane content confirmed the above 60°C finding with broccoli florets:

    “The best treatment temperature for maximizing sulforaphane yield was 60 °C. The slightly higher sulforaphane yield than would be predicted from the level of glucoraphanin in raw broccoli requires further investigation. The sulforaphane yield of broccoli after 5 min thermal treatment at 65 °C was even lower than the value obtained for raw broccoli.”

3. I see socialistic animal behavior often during beach walks. If one seagull pecks a food morsel, a half-dozen others immediately position themselves to take it. It’s a race to the bottom of existence.

Too bad we humans don’t learn pertinent lessons from others’ experiences, much less our own. Today’s US Thanksgiving provides one example.

Richard Ebeling presented the factual Thanksgiving story a while back. Have you read about collectivism that arrived with the Mayflower in 1620? Do you think we’ve learned what we needed to learn about communism from four centuries ago through today?

4. Seagulls are also inspirational in their flock behavior of joie de vivre predawn flying.

Eat broccoli sprouts for your eyes

This 2020 review subject concerned a leading cause of blindness:

“Advanced glycation end products (AGEs) are toxic compounds that have adverse effects on many tissues including the retina and lens. AGEs promote the formation of reactive oxygen species (ROS), which, in turn, boost the production of AGEs, a vicious cycle.

Diabetic retinopathy (DR) is a devastating microvascular complication of diabetes mellitus and the leading cause of blindness in working-age adults. The onset and development of DR is multifactorial. Lowering AGEs accumulation may represent a potential therapeutic approach.

Once AGEs are formed, most are irreversible. Cataracts are perhaps the earliest pathobiology of AGEs:

Nε-(carboxymethyl)-lysine (CML) [a representative AGE] in lens crystallins from diabetic (■) and non-diabetic (♦) subjects as a function of age.

The glyoxalase system is a protective mechanism that slows down synthesis of AGEs by limiting reactive dicarbonyls formed during sugar metabolism. Glutathione (GSH) in the eye is present at concentrations many times blood levels, and is a critical component of the glyoxalase system.

Proteomic analysis identified GLO1 [glyoxalase 1] as a protein differentially expressed in cells treated with sulforaphane. Sulforaphane inhibited AGEs-derived pericyte damage and delayed diabetes-induced retinal photoreceptor cell degeneration.

No AGE inhibitors have reached clinical use. The glyoxalase system and discovery of compounds that enhance this detoxifying activity represent a therapeutic alternative to fight glycation-derived damage.”

https://www.mdpi.com/2076-3921/9/11/1062/htm “Glyoxalase System as a Therapeutic Target against Diabetic Retinopathy”


The above graph – plotting a cataract AGE level against chronological age – represented life stage progression without effective personal agency, without taking responsibility for your one precious life.

Citation 156 was Activation of Nrf2 attenuates carbonyl stress induced by methylglyoxal in human neuroblastoma cells: Increase in GSH levels is a critical event for the detoxification mechanism (not freely available):

“The present study focused on the methylglyoxal (MG) detoxification mechanism. MG treatment resulted in accumulation of modified proteins bearing the structure of AGEs.

This accumulation was suppressed by activation of the Nrf2 pathway prior to MG exposure via pre-treatment with an Nrf2 activator:

Although pre-treatment with the Nrf2 activator did not affect mRNA levels of GLO1, expressions of GCL and xCT mRNA, involved in GSH synthesis, were induced prior to increase in GSH levels.

These results indicated that increase in GSH levels promoted formation of the GLO1 substrate, thereby accelerating MG metabolism via the glyoxalase system and suppressing its toxicity. Promotion of GSH synthesis via the Nrf2/Keap1 pathway is important in MG detoxification.”

Continued in Part 2.


PXL_20201121_113656177

Sulforaphane in the Goldilocks zone

This 2020 paper reviewed hormetic effects of a broccoli sprout compound:

“Sulforaphane (SFN) induces a broad spectrum of chemoprotective effects across multiple organs that are of importance to public health and clinical medicine. This chemoprotection is dominated by hormetic dose responses that are mediated by the Nrf2/ARE pathway and its complex regulatory interactions with other factors and pathways, such as p53 and NF-κB.

The stimulatory zone for in vitro studies proved to be consistently in the 1-10 μM range. Hormetic studies of SFN strongly targeted activation of Nrf2.

Capacity to activate Nrf2 diminishes with age, and may affect capacity of SFN to effectively enhance adaptive responses.

A 4-hour exposure induced a 24 hour Nrf2-mediated increase in enzymes that reduce free-radical damage in neurons and astrocytes. Repeated 4-hour treatment for four days affected an accumulation along with a persistent protection.

In the case of continuous exposure to SFN, such as taking a daily supplement, SFN treatment did not result in an accumulation of HMOX1 [heme oxygenase (decycling) 1 gene] mRNA or protein. This suggested that HMOX1 response may experience feedback regulation, avoiding possible harmful overproduction.”

https://www.sciencedirect.com/science/article/abs/pii/S1043661820315917 “The phytoprotective agent sulforaphane prevents inflammatory degenerative diseases and age-related pathologies via Nrf2-mediated hormesis” (not freely available)


One coauthor has been on a crusade to persuade everybody of this paradigm. Hormesis’ hypothesis isn’t falsifiable in all circumstances, however.

Hormetic effects may be experimental considerations. But what’s the point of performing sulforaphane dose-response experiments in contexts that are physiologically unachievable with humans? Two examples:

  1. Autism biomarkers and sulforaphane:

    “There was no concentration-dependence in the induction of any of the genes examined, with the higher (5 μM) concentration of SF even showing a slightly diminished effect for the induction of AKR1C1 and NQO1. Although this concentration is achievable in vivo, more typical peak concentrations of SF (and its metabolites) in human plasma are 1-2 μM.”

  2. Human relevance of rodent sulforaphane studies:

    “Over two-thirds of the animal studies have used doses that exceed the highest (and bordering on intolerable) doses of sulforaphane used in humans. The greater than 4-log spread of doses used in mice appears to be driven by needs for effect reporting in publications rather than optimization of translational science.”

This paper cited many hormetic effects that were human-irrelevant without making a distinction. It also had parts such as:

“The capacity for high concentrations of AITC [allyl isothiocyanate] to enhance genetic damage is not relevant since such high concentrations are not realistically achievable in normal human activities.

Humans ingest only the R-isomer of SFN via diet. Their dosing strategy adopted concentrations of R-SFN that were less than those employed to induce cytotoxic effects in cancer cells and that simulated its consumption as a dietary supplement.”


Landing eagle

DIM effects on BRCA carriers

This 2020 study evaluated a broccoli sprout compound’s effects on breast cancer development:

“Women who carry the BRCA mutation are at high lifetime risk of breast cancer, but there is no consensus regarding an effective and safe chemoprevention strategy. A large body of evidence suggests that 3,3-diindolylmethane (DIM), a dimer of indole-3-carbinol found in cruciferous vegetables, can potentially prevent carcinogenesis and tumor development.

A year’s supplementation with DIM 100 mg daily in BRCA carriers was associated with a significant decline in FGT [fibroglandular tissue] amount on MRI. Larger randomized studies are warranted to corroborate these findings.”

https://academic.oup.com/carcin/article/41/10/1395/5847633 “3,3-Diindolylmethane (DIM): a nutritional intervention and its impact on breast density in healthy BRCA carriers. A prospective clinical trial”


This study didn’t address DIM bioavailability. What were the DIM amounts each subject actually processed? How was DIM bioavailability related to their “significant decline in FGT” outcome?

Studies that found DIM was only 1-3% bioavailable after oral administration include:

PubChem lists DIM molecular weight as 246.31 g/mol. A 4.06 μmol DIM amount (.001 / 246.31) equals a 1 mg weight. The study’s daily DIM intake 100 mg weight was a 406 μmol amount.

Eat broccoli sprouts for DIM and Our model clinical trial for Changing to a youthful phenotype with broccoli sprouts measured DIM excreted as a result of eating 30 grams raw broccoli super sprouts every day. Indolic glucosinolate precursors of DIM were as follows:

indolic glucosinolates

DIM at the 70-day point was an average 0.650 μmol amount, which was almost twice those subjects’ 0.334 average beginning amount. If each subject’s DIM was collected over 24-hours, using precursor conversion calculations may have produced bioavailability measurements.


Young dolphins eating breakfast

Microwaving broccoli sprouts may not affect phenolic levels

Three papers investigated microwaving plant material for phenolic compounds. The first was a 2020 review that compared industry techniques:

“Phenolic compounds are widely used as natural antioxidants and antimicrobial agents. They also exhibit antiallergenic, antiatherogenic, and anti-inflammatory activities.

Although the nature and properties of raw materials strongly influence extraction, all extraction processes share major parameters:

  • Solubility of phenolic compounds is higher in polar solvents such as water and ethanol or their mixtures;
  • Diffusion of compounds and mass transfer rates are enhanced by increased temperature; and
  • Longer extraction times allow for a more intimate and effective contact between solvent and matrix.

MAE [microwave-assisted extraction] is an efficient technique due to its ability to heat a matrix internally and externally without a thermal gradient. Phenolic compounds and ionic solutions strongly absorb microwave energy. Microwaves cause internal superheating of water molecules of a sample, promoting cellular disruption and enhancing the recovery of target compounds from the matrix.

MAE produces the highest total phenolic content, 227.63 mg GAE [gallic acid equivalent] /g dry basis.”

https://link.springer.com/article/10.1007%2Fs13197-020-04433-2 “Recent advances and comparisons of conventional and alternative extraction techniques of phenolic compounds” (not freely available)


I didn’t see any studies in the first paper that I could directly use. Either temperature was too high, or microwave power too low, or was something I’m not going to do, like substitute ethanol for water.

A second 2020 paper compared three industry techniques to extract phenolics from olive and wine post-processing plant material:

“In the case of olive pomace, TPC [total polyphenol content] improved by increasing the ethanol concentration from 20 to 50%, and temperature from 60 to 90°C. No significant improvement was observed when increasing time from 5 to 15 min.

The best extraction efficiency for olive pomace residues was provided by MAE.”

https://www.mdpi.com/2076-3921/9/11/1074/htm “Olive Mill and Winery Wastes as Viable Sources of Bioactive Compounds: A Study on Polyphenols Recovery”


The second study demonstrated that polyphenols weren’t harmed by temperatures up to 90°C. Microwave power was 500W per correspondence with the authors.

Microwave broccoli to increase sulforaphane levels demonstrated significant differences for 450W (LL) and 900W (HL) power settings in glucoraphanin and sulforaphane amounts when microwaved to the same temperatures. Compare sulforaphane amounts for LL60 and HL60 (both 60°C), annotated as E and F:


A third study from 2017 measured total phenolic contents, but primarily used indole-3-carbinol to probe food preparation methods:

“This study evaluated the effects of grinding and chopping with / without microwaving on the health-beneficial components, and antioxidant, anti-inflammation and anti-proliferation capacities of commercial kale and broccoli samples. The availability of indole-3-carbinol (I3C) and total phenolic contents [TPC] were evaluated.

Fifty grams and 100 g of commercial fresh kale and broccoli (including stem and leaves) samples were ground for 15 s with 200 mL of water in 5 different commercial available blenders. The ground vegetables were divided into two parts, and one part was microwaved at 700 watts for 30 s.

Availability of I3C was significantly enhanced with grinding as compared to chopping (below the limit of detection), suggesting the effect of particle size on food factor release. Particle sizes of ground kale and broccoli were 10 times smaller than chopped counterparts. Grinding [in broccoli] not only resulted in difference of particle size, but might also break plant cell walls and release enzymes such as myrosinase, which might have hydrolyzed glucobrassicin and caused a greater releasable I3C level.

Significant difference was observed in I3C availability with or without microwaving. Microwaving significantly elevated the extractable amount of I3C from broccoli regardless of the blenders used. For instance, availability of I3C in broccoli was increased by 3.1, 9.1 and 1.9 folds respectively using blenders 1, 2 and 5 with microwaving as compared to their unmicrowaved counterparts.

TPC from blended broccoli samples ranged from 0.28-0.47 mg gallic acid g-1 of fresh weight. This range was lower than a reported mean of 0.99 mg gallic acid g-1 of fresh broccoli [in another study], suggesting different cultivars, locations (USA versus France) and extraction methods (water extraction versus 70% acetone extraction) might affect releasable TPC from broccoli.”

https://www.sciencedirect.com/science/article/abs/pii/S0308814617318484 “Home food preparation techniques impacted the availability of natural antioxidants and bioactivities in kale and broccoli” (not freely available)


1. Funny that I found this third paper in a PubMed “microwave phenolic broccoli” search, but not in any Part 2 of Do broccoli sprouts treat migraines? I3C combination searches. A plain “I3C” search term like how I search PubMed weekly on “sulforaphane” would have found it.

2. Don’t understand why blenders 1-5 makes and models weren’t stated in the study. Using blender 1 made a significant difference in TPC in the above graphic, but that was the effect. What could have been the cause? Aren’t researchers obligated to provide such explanations?

And why didn’t the study text support the graphic and address all TPC results with microwaved broccoli? Microwaving produced neither significant TPC differences among blenders 2-5 broccoli samples, nor in any of the kale samples.

3. Also don’t understand why these researchers didn’t microwave chopped broccoli samples and measure them for TPC and I3C. Maybe that wouldn’t have produced anything for TPC if phenolics aren’t produced from the myrosinase hydrolysis chain of events.

But I3C is a myrosinase hydrolysis product. Testing microwaved chopped samples for I3C may have changed the above bolded statement to:

“Microwaving significantly elevated the extractable amount of I3C from broccoli regardless of the blenders used. [food preparation method.]

4. One broccoli treatment was blending 100 grams broccoli in 200 ml water, halving the purée, then microwaving half on 700W power for 30 seconds. The study didn’t say what temperature was achieved, but it was probably < 60°C because that’s similar to what I do.

Twice every day I microwave an average 65.5 grams of 3-day-old broccoli sprouts in 100 ml water on 1000W full power for 35 seconds to achieve up to but not exceeding 60°C. I use the same 100 ml water, but more broccoli sprout weight and microwave wattage to initiate myrosinase hydrolization of glucoraphanin and other glucosinolates into sulforaphane and other healthy compounds.

5. My son encouraged me to try blending microwaved broccoli sprouts this summer. I stopped after two weeks as I consistently had trouble swallowing them.

6. The study design required microwaving broccoli sprouts after blending. I don’t think people would do it in this order at home.

It would be too messy to scrape a broccoli sprout purée out of a blender and into a microwavable dish, while maintaining a desired water volume for microwaving. The reverse order is easier – measure the desired water volume into the broccoli sprouts dish, microwave, then plop microwaved broccoli spouts into a blender.

Broccoli sprout compounds include sinapic acid derivatives

This 2020 study provided details about healthy sinapic acid broccoli sprout compounds:

“Anti-inflammatory effects of broccoli sprouts have been demonstrated in vivo, but the connection with composition is not yet fully explained. The aim of the present study was:

  • Provide a complex qualitative / quantitative insight into variability of SADs [sinapic acid derivatives] during germination of broccoli sprouts; and
  • Investigate their antioxidant and anti-inflammatory properties that might result from the presence of phenolics.

Sprouting in darkness results in overall decrease in total content of SADs with growth time, but promotes replacement of relatively low active constituents, such as sinapine, by stronger antioxidants. These structural changes are beneficial for total antioxidant capacity of broccoli sprouts, and are correlated with their increasing ability to scavenge free radicals, reduce transition metal ions, and inhibit lipid peroxidation.”

The graphic’s dotted line is sinapine.

“Anti-TNF-α activity of broccoli sprout extract and sinapic acid was less relevant than that of an anti-inflammatory drug DEX [dexamethasone]. However, contrary to DEX, they significantly stimulated release of IL-10, which is an anti-inflammatory and immunosuppressive factor. By downregulating secretion of pro-inflammatory cytokines, IL-10 may impede immunopathology by inhibiting activity of macrophages, natural killer cells and lymphocytes.

Most of the previous studies ascribed anti-inflammatory effects of broccoli sprouts to their sulfur compounds only. Research data from in vivo models confirmed that glucosinolates may stimulate IL-10 release while isothiocyanates do not influence significantly or even decrease its production.

The present study indicates that phenolic constituents may also be responsible for anti-inflammatory effects of broccoli sprouts, and stimulation of IL-10 might be most relevant in this context.”

https://pubs.rsc.org/en/content/articlelanding/2020/FO/D0FO01387K “Variability of sinapic acid derivatives during germination and their contribution to antioxidant and anti-inflammatory effects of broccoli sprouts on human plasma and human peripheral blood mononuclear cells” (not freely available)


Some aspects of this study:

1. Seed-2-4-6 day measurements complemented seed-3-5-7 day measurements of the superb 3-day-old broccoli sprouts have the optimal yields. Both studies used standard methodologies, provided convertible dry weight and fresh weight measurements in mg / g, and grew their sprouts in darkness, with the current study watering every 12 hours.

The current study measured composition changes of 31 compounds (18 sinapic acid derivatives, 8 glucosinolates, and 5 flavonoids). It was too complicated for me to sum up and convert these granular dry weight measurements to make them directly comparable with 3-day study total phenolics (TP below, and TF is total flavonoids). They decreased similarly to TP mg / g fresh weight measurements.

The 3-day study also compared each cultivated variety’s germination stage weights and measurements with its origin using a milligram-per-gram-of-seeds scale:

“To be comparable, the content of these bioactive compounds from 100 fresh sprouts was divided by the weight (g) of 100 seeds, and then this value was compared with their content from one gram seeds.

The seed weight of XM was nearly twice that of other varieties and its sprouts weight also had the highest increasing rate. LWW and LY had the lowest weight of seeds as well as sprouts.”

phenolics and flavonoids

Different lowercase letters meant significant differences in the same cultivar among sampling days, and uppercase in the same sampling day among cultivars.

A. That study’s mg / g fresh weight trend favored broccoli seeds, a subject explored in Microwave broccoli seeds to create sulforaphane. Seeds of all six cultivars were significantly higher than their sprouts in TP, TF, and sulforaphane in this paradigm.

B. The mg / g seed trend favored 3-day-old broccoli sprouts for TP (five of six cultivars), TF (all), but not sulforaphane (two of six). Both trends were scientifically informed.

Which paradigm suits you?

2. A 2005 study was cited for:

“Total glucosinolate content in fresh broccoli sprouts were reported to reach 4.02 μmol/g fw, which corresponds to 0.71 mg/g fw of active isothiocyanates. In our study, total content of SADs in 6-day-sprouts was 4.85 mg/g dw. Recalculating to fresh weight gives 0.37 mg/g fw, which suggests that SADs might well influence biological effects of glucosinolates.”

The current study measured eight glucosinolates but no isothiocyanates. Don’t think this 2005 study provided bases for close comparisons, since it had unknown broccoli cultivar, unknown sprout age, and unanalyzed isothiocyanates.

Comparisons to a two-decimal-point precision should use the same cultivar and contexts, to include growing conditions. For example:

  • The 3-day study showed wide variation in sulforaphane weights among six broccoli cultivars’ seeds, from 2.43 mg to 12.07 mg per gram of seeds. And:

    “Knowledge concerning the bioavailability of plant ITCs is essential to predict the potential level of exposure as GL determination alone may not accurately reflect how much of the final active ITC can be formed from sprouts.”

  • The three cultivars studied in Lab analyses of broccoli sprout compounds had broccoli sprout sulforaphane amounts vary from 0.3 to 1.2 μmol / g fw in one growing season, then from 0.2 to 0.6 μmol / g fw the next year.

But these researchers knew that:

“In a study, diminishing amounts of total phenolic acids in sprouts of three broccoli cultivars was observed only between 3rd and 7th day of germination under photoperiod conditions and only when expressed on fresh weight basis. After recalculating results to dry weight, amounts were increasing during the whole 14-day observation period.

Trends cannot be treated as universal, but are dependent on growth conditions and variety of the plant.”

“0.37 mg/g fw” of Day 6 sinapic acid derivatives would be lower than Day 7 of four 3-day study cultivars’ total phenolics, and between Days 5 and 7 of the other two. I’d guess that the current study’s cultivar, Cezar, would be neither the lowest nor the highest cultivar if comparably measured for total phenolics.

3. These researchers compared other studies to an extent that isn’t usually seen. The result was good information such as:

“The most significant difference is sinapine that was not detected in either of the two above mentioned works, but is a major component of extracts investigated in the present work.

In one study, 7-day-sprouts were germinated under photoperiod conditions and without monitoring the phenolic profile in preceding growing days. Dominating constituents identified were trisinapoyl-gentiobiose, that was also present in relevant quantities in 6-day-sprouts, and sinapoyl-malate, that was not detected in our study.

Taking into account the significant role of these compounds (especially sinapoyl-malate) in UV-protection, light availability might have caused much quicker sinapine degradation and synthesis of SADs more suitable for those conditions. The same study also demonstrated that additional UVA and UVB irradiation before harvest further induced production of some of the constituents.

4. Flavonoids and hydroxycinnamic acids are subgroups of phenolic compounds. Sinapic acid derivatives are a category of hydroxycinnamic acids.

Would microwaving sinapic acid derivatives have similar effects to what was described in Microwave broccoli to increase flavonoid levels? That’s the subject of Microwaving broccoli sprouts may not affect phenolic levels.


If it stinks, it’s good for you

This 2019 review subject was hydrogen sulfide and broccoli sprout compounds:

“Release of H2S was identified as a hidden mechanism responsible for effects of natural compounds that were used for a long time for pharmacological, therapeutic or nutraceutical purposes. For instance, the release of H2S was recognized as the main mechanism accounting for the biological effects of garlic.

There is evidence of a close overlap between numerous physiological / biological effects attributed to natural ITCs [isothiocyanates] and H2S. They both behave as antioxidant and anti-inflammatory agents, are activators of potassium channels modulating a vasodilatory effect, and are chemopreventive agents. Moreover, an impressive overlap can be observed in the molecular mechanisms of action.

Vascular inflammation results from the persistence of oxidative and/or inflammatory stimuli on the endothelium and vascular smooth muscle. These types of stimuli can be a consequence of prolonged status of mild inflammation and are typical in certain metabolic / cardiovascular diseases, spreading to all organs and tissues.

Advanced glycation end products (AGEs) are responsible for an increase in LDL. Binding of AGEs to their receptors RAGE results in an increase in intracellular ROS generation and in activation of NF-kB. Oral consumption of SFN [sulforaphane] precursor-rich broccoli sprouts decreases the serum levels of AGEs in humans.

Inflammatory response plays a pivotal role in initiation and maintenance of chronic neuropathic pain. Inhaling low concentrations of H2S protects motor neurons from degeneration and delayed paraplegia in a mouse model of sciatic constriction injury. This effect has been ascribed to the activation of the Nrf2 pathway.

Dose-dependent rise of the pain threshold mediated by SFN was fully prevented by simultaneous administration of hemoglobin, confirming that H2S is likely to be the real player in ITC-induced analgesia. Kv7 channel activation can be considered one of the main mechanisms in the antinociceptive activity of H2S-releasing drugs.”

https://www.liebertpub.com/doi/10.1089/ars.2019.7888 “Organic Isothiocyanates as Hydrogen Sulfide Donors” (not freely available)


These reviewers were long on equivalencies and short on proofs. Unlike study researchers, reviewers aren’t bound to demonstrate evidence from tested hypotheses. Reviewers are free to:

  • Express their beliefs as facts;
  • Over/under emphasize study limitations; and
  • Disregard and misrepresent evidence as they see fit.

Study researchers are obligated to provide detailed analyses of why observed effects couldn’t have been produced from unobserved causes. That didn’t happen here.

Anti-tumor effects of β-glucan

This comprehensive 2020 rodent study investigated dozens of scenarios for β-glucan in the context of anti-tumor immunity:

“Neutrophils and granulopoietic progenitors are major cellular effectors of β-glucan-induced trained immunity. The anti-tumor effect of β-glucan-induced trained immunity was mediated by qualitative changes in neutrophils.

A tumor-suppressive phenotype in neutrophils was associated with training of granulopoiesis mediated by type I IFN [interferon] signaling. Our analysis provided additional evidence for trained immunity-induced epigenetic rewiring of granulopoiesis toward an anti-tumor phenotype and corroborated the experimentally demonstrated IFN- and ROS-related mechanisms.

We observed inhibition of tumor growth by systemic transfer of trained neutrophils into already tumor-bearing mice. As granulocyte transfusion is currently considered as a therapy in humans with neutropenia, it is conceivable that cancer patients could receive as an adjuvant immunotherapy granulocytes from normal donors after induction of trained immunity in the latter.

Our study is the first to link the anti-tumor actions of β-glucan to trained immunity. We show here that the innate immune training and rewiring of granulopoiesis underlies the anti-tumor effect of β-glucan.”

https://www.cell.com/cell/fulltext/S0092-8674(20)31299-X “Innate Immune Training of Granulopoiesis Promotes Anti-tumor Activity”


Which do you prefer? The study’s graphical abstract:

or one of its volcano plots?

Here’s an overview of one investigated direction:

“To determine whether adaptive immunity is involved in the anti-tumor effect induced by β-glucan, mice that lack B and T cells were treated with β-glucan [1 mg] prior to the secondary tumor challenge. Pre-treatment with β-glucan decreased both B16-F10 [melanoma] and LLC [Lewis lung carcinoma] tumor burden also in [these] mice, showing that the anti-tumor effect of β-glucan-induced trained immunity does not require adaptive immunity.”


This study provided another example of what they called rewiring (but I term reprogramming) of the body’s environmental signaling pathways to achieve a desired phenotype, trained innate immunity. Whatever the terminology, almost every day over the past fifteen years I’ve eaten β-glucan in an oats breakfast and a 1/3, 1/6 yeast supplement at dinner as part of individually evolving.

Reprogramming other signaling pathways are in blog posts such as:

Take responsibility for your one precious life.

Flying over waves

Dietary contexts matter

Two papers illustrated how actions of food compounds are affected by their contexts. The first was a 2020 UCLA rodent study:

“Long-chain polyunsaturated fatty acids (PUFAs), particularly omega-3 (n-3) PUFAs, have been indicated to play important roles in various aspects of human health. Controversies are observed in epidemiological and experimental studies regarding the benefits or lack of benefits of n-3 PUFAs.

Dietary docosahexaenoic acid (DHA; 22:6 n-3) supplementation improved select metabolic traits and brain function, and induced transcriptomic and epigenetic alterations in hypothalamic and hippocampal tissues in both context-independent and context-specific manners:

  • In terms of serum triglyceride, glycemic phenotypes, insulin resistance index, and memory retention, DHA did not affect these phenotypes significantly when examined on the chow diet background, but significantly improved these phenotypes in fructose-treated animals.
  • Genes and pathways related with tissue structure were affected by DHA regardless of the dietary context, although the direction of changes are not necessarily the same between contexts. These pathways may represent the core functions of DHA in maintaining cell membrane function and cell signaling.
  • DHA affected the mTOR signaling pathway in hippocampus. In the hypothalamus, altered pathways were more related to innate immunity, such as cytokine-cytokine receptors, NF-κB signaling pathway, and Toll-like receptor signaling pathway.

DHA exhibits differential influence on epigenetic loci, genes, pathways, and metabolic and cognitive phenotypes under different dietary contexts.”

https://onlinelibrary.wiley.com/doi/10.1002/mnfr.202000788 “Multi‐tissue Multi‐omics Nutrigenomics Indicates Context‐specific Effects of DHA on Rat Brain” (not freely available)


A human equivalent age period of the subjects was 12 to 20 years old. If these researchers want to make their study outstanding, they’ll contact their UCLA colleague Dr. Steven Horvath, and apply his new human-rat relative biological age epigenetic clock per A rejuvenation therapy and sulforaphane.

The second paper was a 2016 review Interactions between phytochemicals from fruits and vegetables: Effects on bioactivities and bioavailability (not freely available):

“The biological activities of food phytochemicals depend upon their bioaccessibility and bioavailability which can be affected by the presence of other food components including other bioactive constituents. For instance, α-tocopherol mixed with a flavonol (kaempferol or myricetin) is more effective in inhibiting lipid oxidation induced by free radicals than each component alone.

Interactions of phytochemicals may enhance or reduce the bioavailability of a given compound, depending on the facilitation/competition for cellular uptake and transportation. For example, β-carotene increases the bioavailability of lycopene in human plasma, and quercetin-3-glucoside reduces the absorption of anthocyanins.

Combinations of food extracts containing hydrophilic antioxidants and lipophilic antioxidants showed very high synergistic effects on free radical scavenging activities. A number of phytochemical mixtures and food combinations provide synergistic effects on inhibiting inflammation.

More research should be conducted to understand mechanisms of bioavailability interference considering physiological concentrations, food matrices, and food processing.”


Each of us can set appropriate contexts for our food consumption. Broccoli sprout synergies covered how I take supplements and broccoli sprouts together an hour or two before meals to keep meal contents from lowering sulforaphane bioavailability.

Combinations of my 19 supplements and broccoli sprouts are too many (616,645) for complete analyses. Just pairwise comparisons like the second paper’s example below would be 190 combinations.

binary isobologram

Contexts for each combination’s synergistic, antagonistic, or additive activities may also be influenced by other combinations’ results.

My consumption of flax oil (alpha linolenic acid C18:3) probably has effects similar to DHA since it’s an omega-3 PUFA and I take it with food. The first study’s human equivalent DHA dose was 100mg/kg, with its citation for clinical trials stating “1–9 g/day (0.45–4% of calories) n-3 PUFA.”

A 2020 review Functional Ingredients From Brassicaceae Species: Overview and Perspectives had perspectives such as:

“In many circumstances, the isolated bioactive is not as bioavailable or metabolically active as in the natural food matrix.”

It discussed categories but not combinations of phenolics, carotenoids, phytoalexins, terpenes, phytosteroids, and tocopherols, along with more well-known broccoli compounds.


Diving for breakfast

Rub some broccoli sprouts on it

This 2020 human/rodent study investigated treating and preventing skin photodamage with sulforaphane:

“Alterations in NRF2 signaling have been implicated in aging and stress-induced skin pigmentation disorders in the skin and hair follicles. NRF2 signaling regulates transcriptional programs involved in adaption and survival of cells in the setting of oxidative stress, and oxidative stress occurs in the setting of photodamage.

[1st human experiment with 14 subjects] Expression levels of NRF2 and its target heme-oxygenase-1 (HO-1) were evaluated by immunofluorescence (IF) in skin biopsies. Expression of NRF2 and HO-1 was significantly reduced in skin from individuals >45 years old.

[2nd human experiment with 7 different subjects] The left arm was chosen for treatment with BSE [broccoli sprout extract], as there is typically more photodamage on the left arm due to chronic sun exposure through the car window while driving in the US. A photoprotected area of skin on an upper inner arm was also treated.

Expression of total NRF2 and phosphorylated NRF2 (NRF2-P) by IF microscopy was detected at low baseline levels in photoprotected skin, suggesting some activity of the pathway, whereas the expression of total NRF2 and NRF2-P was undetectable in untreated photoexposed skin (Un). There was significantly elevated IF expression and fold change of IF signal of NRF2 and especially NRF2-P in SF [sulforaphane]-treated skin compared with Un skin in most individuals.

There was no evidence of increased total NRF2 or NRF2-P expression in SF-treated photoexposed skin in 2 individuals. There was also no significant improvement in mottled hyperpigmentation or difference in melanin deposition following SF treatment.

[Six mouse confirmation/exploratory studies] SF is known to have several non-NRF2–mediated targets, such as NF-κB and AP-1. However, our findings suggest that negative regulation of UV-mediated hyperpigmentation observed following SF treatment is occurring in an NRF2-dependent fashion:

  • UVB+SF treatment resulted in more than a 50% decrease in skin pigmentation and melanin deposition, indicating that SF could prevent UVB-induced skin pigmentation.
  • The therapeutic effect of SF on reducing UVB-induced skin pigmentation was dependent on keratinocyte-intrinsic IL-6 receptor α (IL-6Rα) signaling that upregulated NRF2, which led to inhibition of melanogenesis.

Our results provide direct in vivo evidence of how NRF2 is involved in response to oxidative stress associated with photodamage and chronic UV exposure. Treatment of human or mouse skin hyperpigmentation with SF provided the proof of concept for targeting the NRF2 pathway as a therapeutic intervention.”

https://insight.jci.org/articles/view/139342 “Pathogenic and therapeutic role for NRF2 signaling in ultraviolet light–induced skin pigmentation”


Didn’t understand the 2nd experiment’s human dose of 5 nM sulforaphane. The lead author’s cited 2017 study Randomized, split-body, single-blinded clinical trial of topical broccoli sprout extract: Assessing the feasibility of its use in keratin-based disorders used “500 nmol of sulforaphane/mL.” Unless my math is off, the current study and previous study’s doses weren’t equivalent since 1 nM = 0.001 nmol/mL.

I’d like to know more about subjects who didn’t respond to topical sulforaphane treatment. What happened in their lives to make them dead to an evolutionarily-selected antioxidant and anti-inflammatory signaling pathway that influences many other internal environmental signals? Guess we’ll have to wait for:

“Further clinical studies with an increased number of human subjects, longer treatment regimens, and additional body sites are needed to further assess the long-term effects of NRF2 activation on photoaging.”

See Eat broccoli sprouts for your skin! and Eat broccoli sprouts for your hair! for similar studies.


Owl before sunrise

Week 28 of Changing to a youthful phenotype with broccoli sprouts

Did a little math to end this 28th week of eating a clinically relevant weight of microwaved broccoli sprouts every day:

  • I changed the title of weekly updates after Week 7 as a result of A rejuvenation therapy and sulforaphane. Numbers used from its study: “Rats were injected four times on alternate days for 8 days.”
  • Study numbers in Part 2 of Rejuvenation therapy and sulforaphane regarding the new human-rat relative biological age epigenetic clock: “The maximum lifespan for rats and humans were set to 3.8 years and 122.5 years, respectively.” I’m at a similar percentage of species maximum lifespan as were the study’s treated subjects.
  • A human-equivalent multiplication factor that can be applied to a rat post-development time period is 122.5 / 3.8 = 32.2. An 8-day rat treatment period ≈ 258 human days, and 258 / 7 ≈ 37 weeks.

To paraphrase the study’s lead laboratory researcher’s An environmental signaling paradigm of aging paper, aging is a programmed series of life stages. A body clock reset described there and subsequently experimentally tested changed 30 measurements to earlier life stages.

A reset may not require more than what I’ve been doing since the end of March. Maybe 28 weeks hasn’t been long enough to find out?


See the below discussion for a different point of view. I don’t think relative rates of metabolism between species would be more accurate than other measures because of individual differences among humans.

A chart from Microwave broccoli seeds to create sulforaphane of 10 people’s metabolisms after ingesting 200 μmol (35 mg) sulforaphane provides an example. Individual sulforaphane metabolites (DTC is dithiocarbamates) peak plasma measurements ranged from 0.359 μmol to 2.032 μmol.

sulforaphane peak plasma


So we’re patient.

Eat broccoli sprouts to pivot your internal environment’s signals

Two 2020 reviews covered some aspects of a broccoli sprouts primary action – NRF2 signaling pathway activation:

“Full understanding of the properties of drug candidates rely partly on the identification, validation, and use of biomarkers to optimize clinical applications. This review focuses on results from clinical trials with four agents known to target NRF2 signaling in preclinical studies, and evaluates the successes and limitations of biomarkers focused on:

  • Expression of NRF2 target genes [AKR1, GCL, GST, HMOX1, NQO1] and others [HDAC, HSP];
  • Inflammation [COX-2, CRP, IL-1β, IL-6, IP-10, MCP-1, MIG, NF-κB, TNF-α] and oxidative stress [8-OHdG, Cys/CySS, GSH/GSSG] biomarkers;
  • Carcinogen metabolism and adduct biomarkers in unavoidably exposed populations; and
  • Targeted and untargeted metabolomics [HDL, LDL, TG].

No biomarkers excel at defining pharmacodynamic actions in this setting.

SFN [sulforaphane] seems to affect multiple downstream pathways associated with anti-inflammatory actions. NRF2 signaling may be but one pivotal pathway.

SFN is generally considered to be the most potent natural product inducer of Nrf2 signaling. Studies in which these actions are diminished or abrogated in parallel experiments in Nrf2-disrupted mice provide the strongest lines of evidence for a key role of this transcription factor in its actions.

It is equally evident that other modes of action contribute to the molecular responses to SFN in animals and humans. Such polypharmacy may well contribute to the efficacy of the agent in disease prevention and mitigation, but obfuscates the value of specific pharmacodynamic biomarkers in the clinical development and evaluation of SFN.”

https://www.mdpi.com/2076-3921/9/8/716/htm “Current Landscape of NRF2 Biomarkers in Clinical Trials”


Why do researchers still not use epigenetic clocks in sulforaphane clinical trials? Forty mentions of disease in this review, but no consideration of aging?

This was another example of how researchers – even when stuck in a paradigm they know doesn’t sufficiently explain their area (“No biomarkers excel”) – don’t investigate other associated research areas. Why not?

Here’s what Part 2 of Rejuvenation therapy and sulforaphane had to say to those stuck on biomarkers:

“While clinical biomarkers have obvious advantages (being indicative of organ dysfunction or disease), they are neither sufficiently mechanistic nor proximal to fundamental mechanisms of aging to serve as indicators of them. It has long been recognized that epigenetic changes are one of several primary hallmarks of aging.

DNA methylation epigenetic clocks capture aspects of biological age.”


The second review Epigenetic Regulation of NRF2/KEAP1 by Phytochemicals also completely whiffed on epigenetic clocks. One mention of aging in this review, but it wasn’t of:

  • Citation 104 from Archives of Gerontology and Geriatrics; nor of
  • Citation 108 from the March 31, 2020, Aging journal; nor of
  • Citation 131 “Dietary epigenetics in cancer and aging.”

But epigenetic clock and aging associations were certainly in this review’s scope. For example, Citation 119 said:

“Nrf2 transcriptional activity declines with age, leading to age-related GSH loss among other losses associated with Nrf2-activated genes. This effect has implications, too, for decline in vascular function with age. Some of the age-related decline in function can be restored with Nrf2 activation by SFN.”

Why would people bother with phytochemicals (buzzword “compounds produced by plants”) unless to either ameliorate symptoms or address causes?

“Epigenetic Regulation of NRF2/KEAP1 by Phytochemicals” doesn’t occur in just laboratory situations. It’s also part of daily life.

These reviewers were straight-forward with side effects for two of the first review’s four items:

“The best known NRF2 activator that has obtained clinical approval is dimethyl fumarate for the treatment of multiple sclerosis. However, it has several side effects, including allergic reactions and gastrointestinal disturbance. There are a few related agents in clinical trials, such as Bardoxolone and SFX-01, a synthetic derivative of sulforaphane, which also exhibit less than desirable outcomes.”


Human relevance of rodent sulforaphane studies

After reading through findings of several dozen rodent studies this evening, I thought it would be worthwhile to revisit analysis of human relevance provided by one paper of How much sulforaphane is suitable for healthy people?

“Comparisons of published oral doses of sulforaphane administered to mice or rats and sulforaphane (tablets or sulforaphane-rich broccoli preparations) or glucoraphanin-rich broccoli preparations administered to humans.

The allometric scaling of the murine doses uses the correction factor of 0.081 and those for rat doses 0.162. Human doses were based on an estimate of 70 kg body weights in each study.”

A confession followed:

“Animal studies have not delivered all that might be expected of them. Pre-clinical experimentalists have not thought carefully about the selection of dose (or route) and its relevance to clinical utility.

Over two-thirds of the animal studies have used doses that exceed the highest (and bordering on intolerable) doses of sulforaphane used in humans.

Few studies have included a dose-response. The greater than 4-log spread of doses used in mice appears to be driven by needs for effect reporting in publications rather than optimization of translational science.

Authors of this review have contributed to this dose skewing.”


Let’s narrow this graphic to a human-relevant range:

human-rodent-relevant-dosages

48 of the 114 rodent study doses were in an allometric range applicable to humans.

Clinically relevant sulforaphane human doses start at a 100 µmol amount (17.73 mg). The graphic normalized human weights to 70 kg, so 100 µmol / 70 kg is 1.43 µmol / kg. Eyeballing the graphic, 43 of the 114 rodent study doses were in an allometric range applicable to human clinical doses.

But only three of the human sulforaphane study doses were above 4 µmol / kg. This indicator of the mentioned “intolerable doses” will limit clinically relevant oral doses to no more than 17.73 mg x (4 / 1.43) ≈ 50 mg in one serving.

Reviewing clinical trials of broccoli sprouts and their compounds described a sulforaphane study with doses above 4 µmol / kg:

“They proposed the intake of 15 capsules of broccoli sprouts at a time, giving 90 mg of SFN and 180 mg of glucoraphanin, a never before tested dosage, which was established because of the poor life expectancy of the patients and the aggressive characteristics of this type of cancer.

Secondary effects of the chemotherapy, the lack of appetite, nausea, vomits, diarrhea, mouth sores, etc., were factors that made it very difficult for the patients to intake 15 pills at once, as the study initially planned.

Progression of the pancreatic cancer and the GI symptomatology led to a high rate of drop-off of:

  • 72% in the treatment group; and
  • 55% abandonment in the control group!

Therefore, the results were not significant.”

Our model clinical trial Effects of long-term consumption of broccoli sprouts on inflammatory markers in overweight subjects calculated subjects’ mean weight in Table 1 as “85.8 ± 16.7 kg.” Its average glucoraphanin dose per kg body weight was 117 μmol / 85.8 kg = 1.36 μmol / kg.

Per Estimating daily consumption of broccoli sprout compounds, my twice-a-day consumption of a total 131 grams microwaved broccoli sprouts represents a worst-case 52 mg sulforaphane daily intake. This is ≈ 3 µmol / kg, the graphic’s second-largest sulforaphane amount cluster.


Only 9 of the 114 rodent studies were in an allometric range that was both:

  • Clinically relevant to humans as a lower boundary; and
  • Tolerable to humans as an upper boundary.

human-rodent clinically relevant tolerable dosages

The main purpose of animal studies is to help humans. Which researchers conducted sulforaphane studies that could actually help humans?