Take responsibility for your one precious life – Zinc

This 2020 Russian review highlighted clinical data on zinc known before this year:

“Zinc is known to modulate antiviral and antibacterial immunity and regulate inflammatory response. Zinc possesses anti-inflammatory activity by inhibiting NF-κB signaling and modulation of regulatory T-cell functions.

The most critical role of zinc is demonstrated for the immune system. Zinc regulates proliferation, differentiation, maturation, and functioning of leukocytes and lymphocytes.

Alteration of zinc status significantly affects immune response resulting in increased susceptibility to inflammatory and infectious diseases including acquired immune deficiency syndrome, measles, malaria, tuberculosis, and pneumonia. Zn status is associated with the prevalence of respiratory tract infections in children and adults.

In view of the high prevalence of zinc deficiency worldwide (up to 17%), its impact on population health is considered as a significant issue. Certain groups of people, including infants, especially preterm ones, and elderly, are considered to be at high risk of zinc deficiency and its adverse effects.

Zinc was shown to have a significant impact on viral infections through modulation of viral particle entry, fusion, replication, viral protein translation and further release for a number of viruses including those involved in respiratory system pathology. Increasing intracellular Zn levels through application of Zn ionophores significantly alters replication of picornavirus, the leading cause of common cold.

The results of systematic analysis confirmed the efficiency of intake of at least 75 mg/day Zn in reduction of pneumonia symptom duration but not severity, with the response being more pronounced in adults than in children.”

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7255455/ “Zinc and respiratory tract infections: Perspectives for COVID-19”


The review noted a 2014 Spanish rodent cell study which found:

“Labile zinc, a tiny fraction of total intracellular zinc that is loosely bound to proteins and easily interchangeable, modulates the activity of numerous signaling and metabolic pathways. Dietary plant polyphenols such as the flavonoids quercetin and epigallocatechin-gallate act as antioxidants and as signaling molecules.

The activities of numerous enzymes that are targeted by polyphenols are dependent on zinc. The ionophore activity of dietary polyphenols may underlay the raising of labile zinc levels triggered in cells by polyphenols and thus many of their biological actions.”

https://pubs.acs.org/doi/10.1021/jf5014633 “Zinc Ionophore Activity of Quercetin and Epigallocatechin-gallate: From Hepa 1-6 Cells to a Liposome Model” (not freely available)


I get EGCG from drinking 4-5 cups of green tea every day, and 65 mg zinc from supplements. Microwave broccoli to increase flavonoid levels demonstrated 108.5% to 129.8% increases in quercetin and kaempferol levels from microwaving grocery-store broccoli. Microwaving 3-day-old broccoli sprouts may be expected to increase my worst-case calculation of daily 77 mg total flavonoids.

I’ve taken quercetin intermittently per Preliminary findings from a senolytics clinical trial. I’m changing that to take 100 mg quercetin daily.

Autism biomarkers and sulforaphane

This 2020 US human study investigated autism improvements with sulforaphane:

“Autism Spectrum Disorder (ASD) is one of the most common neurodevelopmental disorders that, in the United States, is currently estimated to affect 1 out of 59 children who are 8 years old. Despite decades of research and advances in our knowledge of the etiologies of ASD, treatments and biomarkers for ASD remain limited.

The primary diagnosis of ASD still relies on observational tools that are by nature subjective. There are currently no drugs approved to treat the core symptoms of ASD, nor are there any studies using SF [sulforaphane] in genetic mouse models of ASD.

In our previous placebo-controlled, double-blinded, randomized clinical trial, daily administration of SF for 4-18 weeks substantially improved the behavioral abnormalities of the majority of 26 young males with moderate to severe ASD without significant toxicity. The multi-functional phytochemical sulforaphane affects many of the biochemical abnormalities associated with ASD.

We investigated potential molecular markers from three ASD-associated physiological pathways that can be affected by sulforaphane:

  1. Redox metabolism / oxidative stress;
  2. Heat shock response; and
  3. Immune dysregulation / inflammation

in peripheral blood mononuclear cells (PBMCs) from healthy donors and patients with ASD.

Three representative Nrf2 [nuclear factor erythroid 2-related factor 2]-dependent enzymes:

  1. AKR1C1 [aldo-keto reductase family 1 member C1];
  2. NQO1 [dehydrogenase quinone 1]; and
  3. HO-1 [heme oxygenase]

were significantly induced by 6 h of 2 μM or 5 μM SF ex vivo treatments in PBMCs from healthy donors. This time point was chosen based on our earlier observations of the kinetics of upregulation of Nrf2-dependent genes by SF, and was expected to capture the increased mRNA production of both very fast (HO-1) and relatively slow (NQO1) responders.

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.

SF ex vivo pre-treatment significantly decreased the LPS [lipopolysaccharides]-stimulated inflammatory gene (

  • COX-2,
  • TNF-α,
  • IL-6 and
  • IL-1β

) expression levels in PBMCs from healthy donors.

As a pilot study for a clinical trial of SF in children with ASD, we evaluated the same biomarkers from the ex vivo studies in 10 young males with ASD, 6-12 years of age, who received SF (in the form of a dietary supplement containing GR [glucoraphanin] and myrosinase), 2.2 μmol/kg/d for 14 days. Grouping by broad functionality (e.g. cytoprotective or pro-inflammatory), differences from baseline were highly significant.

asd gene expression

Individually none is sufficiently specific or sensitive, but when grouped by function as two panels, these biomarkers show promise for monitoring pharmacodynamic responses to sulforaphane in both healthy and autistic humans, and providing guidance for biomedical interventions. We conducted this study in the context of ASD, however our findings have broader implications and suggest that these biomarkers can be used in any study involving an intervention with SF.

Major signaling pathways for protective mechanisms against ASD by SF:

  • (a) Keap1/Nrf2/ARE pathway,
  • (b) NF-κB inflammatory pathway,
  • (c) Heat-shock responses.”

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7118069/ “Biomarker Exploration in Human Peripheral Blood Mononuclear Cells for Monitoring Sulforaphane Treatment Responses in Autism Spectrum Disorder”


Broccoli sprouts and sulforaphane aren’t panaceas. Their research is becoming more intensive and focused, though.

Microwave broccoli seeds to create sulforaphane

Two sulforaphane topics came up in discussions with my wonderful woman. Our first was an inference:

  1. 3-day-old broccoli sprouts have the optimal yields found that broccoli sprout sulforaphane content (after processing for analysis) ranged from 46% to 97% of broccoli seeds.
  2. Microwave broccoli to increase sulforaphane levels found that microwaving broccoli florets to 60°C (140°F) increased the sulforaphane amount from .22 to 2.45 µmol / g (1,114%!!).
  3. Wouldn’t broccoli seeds’ sulforaphane be more than broccoli sprouts by microwaving seeds up to 60°C in the same amount of water?

The 3-day study broccoli sprout measurements were relative to each variety’s seeds:

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

Broccoli compounds are similar among broccoli florets, sprouts, and seeds. A major difference is that broccoli sprouts and seeds have no initial sulforaphane content because hydrolization hasn’t occurred yet. The above graphic’s seed and sprout sulforaphane content was created by processing for analysis.

I’ll reason that sulforaphane would be created by:

  • Microwaving one tablespoon of broccoli seeds with a 1000W microwave in 100 ml of distilled water for 30 seconds to achieve up to 60°C; then
  • Straining out the water; then
  • Allowing further myrosinase hydrolization of glucoraphanin and other glucosinolates into sulforaphane and other healthy compounds.

Broccoli seeds are dry, and microwaving acts directly on a material’s water content. The 3-day study methods “immersed [broccoli seeds] in distilled water and soaked at 30°C for 2 h” to start germination. I’ll stipulate two hours as a minimum broccoli seed soaking time before microwaving.

I’ve tried microwaving broccoli seeds five times so far to see if they’re palatable. Seeds soaked for at least two hours then microwaved for 30 seconds swell to almost twice their dry size. They’re easier to strain, chew thoroughly to ensure hydrolization, and swallow.

The 3-day study also found “total phenolic and flavonoid contents in sprouts were 1.12 to 3.58 times higher than seeds.” I won’t stop eating broccoli sprouts, but sometimes it may be expedient to reduce a 72-hour preparation time to 2 hours and still benefit from sulforaphane and other healthy broccoli compounds.

Let’s use Estimating daily consumption of broccoli sprout compounds runt-of-the-litter calculations and assumptions to make a worst-case estimate of sulforaphane content in one tablespoon of broccoli seeds:

  • Minimum broccoli seed weight of one tablespoon (2,436 seeds / 100) x .33 g = 8.04 g.
  • Minimum sulforaphane weight in one tablespoon of broccoli seeds (8.04 g x 2.43 mg sulforaphane per gram of seeds) = 19.54 mg.

I won’t calculate a worst-case sulforaphane weight after microwaving because part of the 3-day study processing for analysis was:

“Broccoli seeds were comminuted by analysis grinder. Seed powder (0.5g) was immersed in distilled water at 55 °C for 5 min to inactivate the epithiospecifier protein.”

Grinding seeds into powder then heating it probably incorporates any effects of microwaving intact broccoli seeds up to 60°C.


Our second discussion topic came by gathering study data from Broccoli or Sulforaphane: Is It the Source or Dose That Matters?

Assessing these 200 μmol amount / 35 mg weight sulforaphane supplement dose studies:

  1. Peak plasma statistics ranged from 0.5 μmol in Row 2 (n = 20) to 2.15 (n = 4) μmol in Row 1. Row 4 (n = 10) statistics don’t show it, but its individual peak plasma ranges per the below graphic were 0.359 μmol to 2.032 μmol. Coincidentally, the Row 4 subject (#2) who had the lowest peak plasma amount also had the lowest urinary % of dose excreted (also termed bioavailability) of 19.5%, and the Row 4 subject (#8) who had the highest peak plasma amount also had the highest sulforaphane bioavailability of 86.9%.
  2. From the Row 4 study: “The half-life of SF in the body was 2.07 ± 0.26 h as calculated from serum area-under-the-curve determinations.” Its Subject #2 had the longest sulforaphane half-life at 2.709 hours.
  3. The peak time after dose ranged from 1 to 3 hours. Not sure why Row 4 didn’t calculate a peak time, but eyeballing the above graphic showed that all subjects peaked between 1 and 2 hours. Row 2’s time was at the study’s first of three measurement intervals (3, 6, and 12 hours). Its peak time after dose probably also took place between 1 and 2 hours.

These four studies showed that there’s wide variation among individual responses to sulforaphane supplements. Row 4 study’s Concluding Remarks ended with:

“Innate metabolic differences must not be discounted when assessing the metabolism of SF alone, delivered in supplements.”


The first of A pair of broccoli sprout studies was Row 2 (n = 20) above. Its sulforaphane supplement statistics – repeated in the below graphic’s BSE (broccoli sprout extract) column – demonstrated how humans’ sulforaphane supplement metabolic profiles were different than our fresh broccoli sprout metabolic profiles:

The divided dose was twelve hours apart at breakfast and dinner times. Also, its first measurements weren’t taken until 3 hours after ingesting, which explains its later times with lesser amounts than the above sulforaphane supplement studies’ earlier times with greater amounts.

I changed my practices to eat microwaved broccoli sprouts at breakfast and dinner times from its finding:

“In sprout consumers, plasma concentrations were 2.4-fold higher after consuming the second dose than after the first dose.”

A metabolic profile resulting from my current practices is probably between the Sprout and BSE divided-dose statistics:

  • Sulforaphane intake is greater than eating raw broccoli sprouts because microwaving 3-day-old broccoli sprouts creates an increased amount of sulforaphane in them before eating.
  • Sulforaphane uptake from microwaved broccoli sprouts is quicker than eating raw broccoli sprouts. It may not be as immediate as taking sulforaphane supplements, which are usually powders.
  • Sulforaphane dose from microwaved broccoli sprouts is less dependent on an individual’s metabolism than eating raw broccoli sprouts.
  • Sulforaphane release from microwaved broccoli sprouts probably continues on to the gut as does eating raw broccoli sprouts. Sulforaphane release from supplements typically ends in the stomach.

The microwaving study processed 10 grams of broccoli florets immersed in 500 ml water with a 950W microwave on full power for 108 seconds to achieve 60°C. I microwave a worst-case 38 grams of 3-day-old broccoli sprouts immersed in 100 ml water with a 1000W microwave on full power for 35 seconds to achieve 60°C.

After microwaving I transfer broccoli sprouts to a strainer, and wait five minutes to allow further myrosinase hydrolization of glucoraphanin and other glucosinolates into sulforaphane and other healthy compounds. Enhancing sulforaphane content provided evidence that myrosinase hydrolization peaks at one minute after achieving 60°C per the below graphic:

I interpret the above sulforaphane degradation from minutes 1 to 5 to be leaching caused by leaving the broccoli sample immersed in water. I strain water from broccoli sprouts after microwaving – the Time 0 mark of the above graphic – because without leaching water, further hydrolization may increase sulforaphane.


Sulforaphane supplements:

  • Are readily metabolized,
  • Blood plasma levels peak by two hours, and
  • Blood plasma levels dissipate by eight hours.

To the extent a metabolism resulting from my current practices is closer to a sulforaphane supplement profile than a raw broccoli sprouts profile, maybe that leaves the door open to a microwaved broccoli seed dose at lunch time? My beautiful woman thinks so. What do you think?

Measuring sulforaphane plasma compounds

This 2020 Australian human study investigated methods of measuring sulforaphane plasma compounds:

“A simplified methodology to allow high-throughput LC–MS [Liquid Chromatography-Mass Spectrometry] analysis of plasma samples for the measurement of sulforaphane and its metabolites is described. Analysis time is greatly reduced by employing fast chromatography and simple plasma extraction procedure.”

“The participants were observed consuming four Broccomax capsules, each containing 30 mg of broccoli seed extract and a dose of 8 mg of sulforaphane, as per manufacturer certificate of analysis, resulting in a total dose of 32 mg of sulforaphane (120 mg of broccoli seed extract).

The mean peak of combined metabolites from our study (0.9 and 1 μM) using 120 mg of broccoli seed extract (~32 mg of SFN) was similar to work by Fahey et al. who investigated the pharmacokinetics of 350 mg of purified broccoli seed powder (mean 1.3 μM ± 0.5 μM), though our dose was almost three-times less. The pharmacokinetic profiles of our study mirrored those of Fahey et al. in that excretion was complete 8 hrs after consumption. Our intervention peaked slightly later (~2hrs), than that of Fahey (~1 hr), likely due to our use of a capsule rather than liquid.”

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7070302/ “Measuring Sulforaphane and Its Metabolites in Human Plasma: A High Throughput Method”


The study was thin on comparing their 2-person results to previous work. I filled in other comparables from Broccoli or Sulforaphane: Is It the Source or Dose That Matters?


The current study set up a strawman by stating a false comparison:

“Our dose was almost three-times less.”

The compared study was the n = 10 subjects row above, which stated its dose as:

“200 μmol of SF was contained in about 350 mg of SF-αCD powder dissolved in 25 mL of distilled water, which subjects were given to drink upon arrival at the clinic.”

If the current study wanted a true comparison, they would measure and compare sulforaphane dose weights or amounts:

  • https://pubchem.ncbi.nlm.nih.gov/compound/sulforaphane lists sulforaphane’s molecular weight as 177.3 g / mol.
  • A 5.64 μmol sulforaphane amount (.001 / 177.3) equals a 1 mg weight of sulforaphane.
  • 200 μmol / 5.64 μmol = 35 mg sulforaphane used in the compared study.

But these researchers couldn’t even do that. They asserted a 32 mg sulforaphane dose “per manufacturer certificate of analysis” when they had the resources to do otherwise.

Why would a study that went to all the trouble of measuring sulforaphane not test their process by measuring their dose? Had they closely read the compared study, they may have also noticed that its commercial supplement, Prostaphane, was tested to verify stated dosage.

Microwave broccoli to increase flavonoid levels

This 2019 USDA study investigated representative broccoli cooking methods for their impact on kaempferol and quercetin levels:

“Understanding cooking effects on flavonoids is crucial to accurately estimate their daily intake and further investigate their health benefits. The purpose of this study was not to compare different conditions of each cooking methods, but to focus on retentions of the individual flavonoids under common cooking conditions in the US:

  • For boiling, we chose to use 85 seconds.
  • A 5-minute steaming time was used.
  • Microwave treatment was carried out in a 1200W microwave at full power for 1 minute.

Seven kaempferol (Km) glycosides and one quercetin (Qn) glycoside were identified and quantified in raw and cooked broccoli by HPLC-MS:

Boiling resulted in significant loss of all flavonoids, while steaming and microwaving led to minor losses or even increases of the flavonoids.

Microwaving without water or with small amount of water tended to retain or increase total phenolics and/or flavonoids. When a large amount of water was added during microwaving, to some extent resembling boiling, total phenolics and/or flavonoids decreased.

Different agriculture practices may affect flavonoids’ existence and their interactions with other compounds, which in turn alter their sensitivity to heat treatments.

Retention of nutrients in cooked foods can be calculated as apparent retention (AR) based on dry form, or true retention (TR) based on fresh/wet form. TR represents the actual consumption forms and also takes the weight change after cooking into consideration.

The possible explanation for TR over 100% is that the thermal processing may increase the extractability and/or the release from binding to other compounds as a result of matrix softening.”

https://www.cell.com/heliyon/fulltext/S2405-8440(19)30568-7 “Effects of domestic cooking on flavonoids in broccoli and calculation of retention factors”


The Material and methods section didn’t state that heated products’ temperatures were measured. So there wasn’t sufficient evidence for a solely thermal explanation of only microwaving achieving percentages over 100 per:

“The possible explanation for TR over 100% is that the thermal processing..”

A more plausible explanation similar to Microwave broccoli to increase sulforaphane levels may account for microwaving’s increased percentages:

“Microwave treatment causes a sudden collapse of cell structure due to the increase in osmotic pressure difference over vacuole membrane.

We didn’t expect this result, and think microwave irradiation might help to release more conjugated forms of glucosinolates and then get hydrolyzed by released myrosinase.”


I immerse 3-day-old broccoli sprouts in 100 ml distilled water, then microwave them on 1000W full power for 35 seconds to achieve up to but not exceeding 60°C (140°F). After microwaving I transfer broccoli sprouts to a strainer, and allow further myrosinase hydrolization of glucoraphanin and other glucosinolates into sulforaphane and other healthy compounds.

Myrosinase deactivation above 60°C apparently wasn’t a consideration, since boiling, steaming, and a 1200W microwave on full power for one minute may have produced temperatures above 60°C. I’ll guess that an active enzyme wasn’t a requirement for flavonoid contents of broccoli purchased in a Beltsville, Maryland, grocery store.

The microwave tests used:

“Broccoli florets (150 g) were put in a microwave safe bowl with a 1 tablespoon [15 ml] of water.”

I use:

  • A lesser weight of 3-day-old broccoli sprouts;
  • A greater volume of distilled water;
  • A less powerful microwave operated on full power for a lesser duration.

Before microwaving, I would expect a worst-case estimated 77 mg total flavonoids from eating 3-day old broccoli sprouts every day. This study’s findings lead me to expect that current practices with microwaving would improve flavonoid levels.

Don’t overcook broccoli

This 2020 US / Korea study set a low bar and jumped over it by finding:

“The abundance of GSL [glucosinolate] hydrolysis products in cooked samples was lower compared to the raw samples.

Regardless of different cooking methods and durations, the total GSL amount in MeJA [methyl jasmonate]-treated broccoli was still higher than in the non-treated broccoli. This suggests that the increased GSL concentration in broccoli samples was solely affected by MeJA treatment, and the effect of MeJA was not affected by cooking methods.

Effect of cooking and 250 µM MeJA treatment on (A) total aliphatic glucosinolates, (B) total indole glucosinolates, and (C) total glucosinolates in ‘Green Magic’ broccoli. * = detected significant different by Student’s T-test (p ≤ 0.05, n = 3) with a significant interaction between MeJA treatment and cooking treatment.”

https://www.mdpi.com/2304-8158/9/6/758/htm “Methyl Jasmonate Treatment of Broccoli Enhanced Glucosinolate Concentration, Which Was Retained after Boiling, Steaming, or Microwaving”


Did it advance science to only replicate mistakes in consumer broccoli cooking methods with:

“The abundance of GSL hydrolysis products in cooked samples was lower compared to the raw samples.”

No.

Did the study design have tests to provide cooking method guidance for:

“To date, methods of delivering cooked broccoli without losing its nutritional benefits are still lacking in the literature, although consuming cooked broccoli is the most common practice for consumers.”

No.

Were there cooking method and temperature recommendations to avoid:

“Cooking also inactivates myrosinase, the enzyme converting GSL into hydrolysis products, and then hinders the formation of hydrolysis products.”

No.

Were there cooking method tests to further enhance either control samples or:

“Exogenous methyl jasmonate (MeJA) treatment was known to increase the levels of neoglucobrassicin and their bioactive hydrolysis products in broccoli.”

No.

Why omit temperature measurements since:

“The major research questions of this study were to evaluate how MeJA application to broccoli plants will affect GSL concentration, myrosinase activity, GSL hydrolysis product amounts..”

Maybe Microwave broccoli to increase sulforaphane levels wasn’t yet published when this study’s design decisions were made. Still, why would a study:

  • Test microwave half power without also testing full power?
  • Select microwaving time as the sole measurement without also measuring temperature?

Table S1 and Figure 3 of the Chinese / USDA study showed a two-minute microwaving time at 50% power wouldn’t be expected to have any sulforaphane content significantly different from uncooked broccoli. Also, temperatures of a five-minute microwaving time at 50% power were guaranteed to completely deactivate myrosinase.

Supplementary material confirmed that this study’s microwaving parameters didn’t show anything of value for how to use your microwave to increase broccoli compound levels. Did the study’s findings provide much more than what not to do?

Poor design decisions created a large gap between what could have been studied and what was studied. Let’s hope there will be better use of resources next time.

Part 2 of Do broccoli sprouts treat migraines?

To follow up Do broccoli sprouts treat migraines? which used a PubMed “sulforaphane migraine” search, a PubMed “diindolylmethane” search came across a 2020 Czech human cell study Antimigraine Drug Avitriptan Is a Ligand and Agonist of Human Aryl Hydrocarbon Receptor that Induces CYP1A1 in Hepatic and Intestinal Cells that had this informative Introduction:

“The aryl hydrocarbon receptor (AhR) transcriptionally controls a wide array of genes. AhR is a critical player in human physiology (e.g., hematopoiesis) and also in many pathophysiological processes such as diabetes, carcinogenesis, inflammation, infection or cardiovascular diseases.

Suitable candidates for off-targeting AhR could be the antimigraine drugs of triptan class, which have an indole core in their structure. Indole-based compounds were demonstrated as ligands of AhR, including dietary indoles (e.g., indole-3-carbinol and diindolylmethane).”

Adding AhR to the search showed:

Changing the PubMed search to “icz migraine” pulled up a 2013 review Biomedical Importance of Indoles that described sumatriptan as an indole, and:

“Since DIM accumulates in the cell nucleus, it likely contributes to cell nuclear events that have been ascribed to I3C.”

Widening the search to “i3c ahr” added:

Changing the search to “i3c migraine” picked up a 2011 UK human study Effect of diindolylmethane supplementation on low-grade cervical cytological abnormalities: double-blind, randomised, controlled trial:

“In the study reported here, there was no statistically significant difference in serious adverse events between groups; in fact a higher proportion of women in the placebo group reported a serious adverse event. Although this study did not have sufficient power to study migraines, we did find a non-significant increase in reported headaches (18% on DIM, 12% on placebo, P=0.12).”

Returning to the original PubMed “sulforaphane migraine” search, Bioavailability of Sulforaphane Following Ingestion of Glucoraphanin-Rich Broccoli Sprout and Seed Extracts with Active Myrosinase: A Pilot Study of the Effects of Proton Pump Inhibitor Administration included one subject who took migraine medication. They weren’t a study outlier, however.


Although indole chemistry indicates a broccoli sprouts – migraine connection, I haven’t found relevant research. Maybe the known properties and actions of broccoli sprout compounds provide enough to affect causes of migraines?

Eat broccoli sprouts for DIM

This 2019 Spanish human study ran in parallel with Our model clinical trial for Changing to a youthful phenotype with broccoli sprouts. I’ll focus on the aspect of diindolylmethane (DIM) from eating broccoli sprouts:

“The aim of this study is to evaluate the effect of gender or hormonal status (menopause) on the bioavailability of broccoli sprouts in different cohorts of overweight adult subjects: men, non-menopausal women and post-menopausal women.

3,3′-diindolylmethane (DIM) was detected and quantified in all volunteers. It increased significantly during broccoli [sprouts] ingestion in men. However, a steady decrease of its urinary concentration was observed in post-menopausal women that was significant at day 50. No significant changes were observed in premenopausal women. Albeit this different behaviour, no significant differences between the three groups were detected by the different statistical tests performed.

High increases observed in SFN-metabolites in the three cohorts confirm that the fresh product is a good source of bioactive compounds bioavailable in the organism. We detected high amounts of 3,3-DIM in urine samples, which can be related to the metabolism of glucobrassicin derivatives from our broccoli sprouts.

Post-menopausal women seem to metabolize isothiocyanates in a greater extension. Hormonal status and differences in gut microbiota may influence the bioavailability of isothiocyanates from broccoli sprouts but more studies are needed to support this statement.”

https://www.sciencedirect.com/science/article/abs/pii/S1756464619303147 “Bioavailability of broccoli sprouts in different human overweight populations” (not freely available)


“Post-menopausal women seem to metabolize isothiocyanates in a greater extension. A steady decrease of its [DIM] urinary concentration was observed in post-menopausal women that was significant at day 50.”

Subjects ate broccoli sprouts every day through Day 35, then stopped, and were measured again at Day 50. The only example of measurements where Day 35 was less than Day 0 was postmenopausal women retaining more broccoli sprout indolic compounds’ metabolite, DIM, than excreting it.

That Day 35 data point didn’t have an asterisk next to it to indicate a statistically significant decrease. But the group’s next Day 50 significant “steady decrease” finding supported an interpretation that eating broccoli sprouts supplied those overweight postmenopausal women with DIM that they especially needed.

Regarding the huge percentage changes above, our model clinical trial found in a longer time frame:

The decrease in IL-6 levels was significantly related to the increase in 24 h urine SFN [sulforaphane] levels. In case of C-reactive protein, the decrease was significantly related to the increases in 24 h urine SFN-NAC [SFN-N-acetylcysteine] and SFN-CYS [SFN-cysteine].

I’ll guess that these parallel trial subjects also experienced similar benefits from eating broccoli  sprouts every day for five weeks. See Day 70 results from Changing to a youthful phenotype with broccoli sprouts for another guess that even shorter time frames would be effective.


Broccoli sprout indolic compounds that metabolize to DIM:

Day 70 results from Changing to a youthful phenotype with broccoli sprouts

Here are my Day 70 measurements* to follow up Our model clinical trial for Changing to a youthful phenotype with broccoli sprouts, which had these findings:


Keep in mind that I’m not in the population represented by the clinical trial sample:

  1. My chronological age is above their inclusion range;
  2. My BMI is below their inclusion range; and
  3. I take supplements and meet other exclusion criteria.

I also didn’t take Day 0 measurements.

June 2019 BMI: 24.8

June 2020 BMI: 22.4

2020 IL-6: 1.0 pg / ml. See Part 2 of Rejuvenation therapy and sulforaphane for comparisons.

2020 C-reactive protein: < 1 mg / l.

2019 and 2020 No biological age measurements. Why aren’t epigenetic clocks standard and affordable?


I’ve made four lifestyle “interventions” since last summer:

  1. In July 2019 I started to reduce my consumption of advanced glycation end products after reading Dr. Vlassara’s AGE-Less Diet: How a Chemical in the Foods We Eat Promotes Disease, Obesity, and Aging and the Steps We Can Take to Stop It.
  2. In September I started non-prescription daily treatments of Vitamin D, zinc, and DHEA per clinical trial Reversal of aging and immunosenescent trends.
  3. Also in September, I started non-prescription intermittent quercetin treatments of Preliminary findings from a senolytics clinical trial.
  4. I started eating broccoli sprouts every day eleven weeks ago.

1. Broccoli sprouts oppose effects of advanced glycation end products (AGEs) provided examples of Items 1 and 4 interactions.

2. Two examples of Item 2 treatment interactions with Item 4 are in Reversal of aging and immunosenescent trends with sulforaphane:

  • “The effects of the combined treatment with BSE [broccoli sprout extract] and zinc were always greater than those of single treatments.”
  • “Vitamin D administration decreased tumor incidence and size, and the co-administration with SFN [sulforaphane] magnified the effects. The addition of SFN decreased the activity of histone deacetylase and increased autophagy.”

3. How broccoli sprout compounds may complement three supplements I take was in a 2020 review Central and Peripheral Metabolic Defects Contribute to the Pathogenesis of Alzheimer’s Disease: Targeting Mitochondria for Diagnosis and Prevention:

“The nutrients benefit mitochondria in four ways, by:

  • Ameliorating oxidative stress, for example, lipoic acid;
  • Activating phase II enzymes that improve antioxidant defenses, for example, sulforaphane;
  • Enhancing mitochondrial remodeling, for example, acetyl-l-carnitine; and
  • Protecting mitochondrial enzymes and/or stimulating mitochondrial enzyme activities, for example, enzyme cofactors, such as B vitamins and coenzyme Q10 .

In addition to using mitochondrial nutrients individually, the combined use of mitochondrial nutrients may provide a better strategy for mitochondrial protection.”

The review provided a boatload of mitochondrial multifactorial analyses for Alzheimer’s. But these analyses didn’t include effective mitochondrial treatments of ultimate aging causes. I didn’t see evidence of why, after fifteen years of treating mitochondrial effects with supplements, treating one more effect could account for my Week 9 vastly different experiences.


I nod to An environmental signaling paradigm of aging explanations. Its Section 10 reviewed IL-6, C-reactive protein, senescence, and NF-κB in terms of feedback loops, beginning with:

“It is clear that the increasing number of senescent cells depends on the post-adult developmental stage rather than chronological age. The coincidence that these processes result in particular forms of impairment in old age does not seem to be random as it is present in all mammals, and may be causative of many aspects of aging.”

A derived hypothesis: After sufficient strength and duration, broccoli sprout compounds changed my signaling environment, with appreciable effects beginning in Week 9.

I offered weak supporting evidence in Upgrade your brain’s switchboard with broccoli sprouts where a study’s insufficient one week duration of an insufficient daily 17.3 mg sulforaphane dosage still managed to change a blood antioxidant that may have changed four thalamus-brain-area metabolites. For duration and weight comparisons, I doubled my daily amount of broccoli seeds from one to two tablespoons just before Week 6 (Day 35), and from that point onward consumed a worst-case estimated 30 mg sulforaphane with microwaving 3-day-old broccoli sprouts every day.

Maybe a promised “In a submitted study, we will report that peripheral GSH levels may be correlated with cognitive functions” will provide stronger evidence? I’m not holding my breath for relevant studies because:

  • There wouldn’t be potential payoffs for companies to study any broccoli sprout compound connections with research areas such as aging, migraines, etc. Daily clinically-relevant broccoli sprout dosages can be grown for < $500 a year.
  • Sponsors would have to change paradigms, a very-low-probability event. They’d have to explain why enormous resources dedicated to current frameworks haven’t produced effective long-term treatments.

What long-term benefits could be expected if I continue eating broccoli sprouts every day?

The longest relevant clinical trial I’ve seen – referenced in Part 2 of Reversal of aging and immunosenescent trends with sulforaphane – was twelve weeks. Part 2 also provided epigenetic clock examples of changes measured after 9 months, which accelerated from there to the 12-month end-of-trial point.

Reviewing clinical trials of broccoli sprouts and their compounds pointed out:

“Biomarkers of effect need more time than biomarkers of exposure to be influenced by dietary treatment.”


A contrary argument: Perhaps people don’t require long durations to effectively change their signaling environments?

I apparently didn’t start eating an effective-for-me daily broccoli sprouts dosage until Day 35, when I changed from one to two tablespoons of broccoli seeds a day. If so, Weeks 6 through 8 may account for my substantial responses during Week 9.

Could eating broccoli sprouts every day for four weeks dramatically change a person’s signaling environment?

Do you have four weeks and $38 to find out? Two tablespoons of broccoli seeds = 38 g x 30 days = 1.14 kg or 2.5 lbs.

This is what twice-a-day one-tablespoon starting amounts of broccoli seeds look like through three days:


Maintaining the sprouting process hasn’t been a big effort compared with the benefits.

In the absence of determinative evidence, I’ll continue eating broccoli sprouts every day. Several areas of my annual physical have room for improvements. Extending my four lifestyle “interventions” a few more months may also provide hints toward inadequately researched connections.

* Results may not be extrapolatable to other people, to any specific condition, etc.

Our model clinical trial for Changing to a youthful phenotype with broccoli sprouts

The further I get into a daily regimen of eating broccoli sprouts for ten weeks, the more I appreciate “Effects of long-term consumption of broccoli sprouts on inflammatory markers in overweight subjects.”

“This study represents an advance in intervention studies as the broccoli sprouts were included in a daily dietary pattern in quantities that reflect a real consumption. The hypothesis of our research is that broccoli sprouts are able to reduce the inflammatory status in overweight subjects due to their content in phytochemicals, mainly glucosinolates.

Total concentration of aliphatic glucosinolates was 80.50 mg/30 gf.w. This concentration was two-fold higher than indolic glucosinolates. Volunteers consumed an average of 51 mg (117 μmol) and 20 mg (42 μmol) of glucoraphanin and neoglucobrassicin, respectively, on a daily basis, during the 70 days of the dietary intervention. Considering an amount of GRA [glucoraphanin] of 117 μmol by serving, a 4% on average was metabolized through mercapturic acid pathway.

No significant changes were observed in weight and BMI. By contrast, body fat mass slightly decreased significantly after 70 days of broccoli [sprout] consumption and returned to basal levels at day 90, a state that was maintained until day 160.

The decrease in IL-6 levels was significantly related to the increase in 24 h urine SFN [sulforaphane] levels. In case of C-reactive protein, the decrease was significantly related to the increases in 24 h urine SFN-NAC [SFN-N-acetylcysteine] and SFN-CYS [SFN-cysteine].

The possible synergistic interaction of both SFN and 3,30-DIM and the isothiocyanates erucin and sulforaphane are interconvertible, so that the anti-inflammatory effects observed with broccoli sprouts intake are likely due to the combined effects of all the hydrolysis products of glucosinolates.

https://www.sciencedirect.com/science/article/abs/pii/S0261561418301183 (Not freely available, better format) and https://researchonline.lshtm.ac.uk/id/eprint/4647168/ (freely available)


Modifications I’ve made to the clinical trial’s protocols include:

  1. I start new broccoli sprout batches twice a day with one tablespoon of seeds per A pair of broccoli sprout studies.
  2. Per 3-day-old broccoli sprouts have the optimal yields, I consume broccoli sprouts when they’re 3 days old. The clinical trial subjects ate broccoli sprouts that were at least a week old.
  3. I immerse 3-day-old broccoli sprouts in 100 ml distilled water, then microwave them on 1000W full power for 35 seconds to achieve up to but not exceeding 60°C (140°F) per Microwave broccoli to increase sulforaphane levels.
  4. Per Enhancing sulforaphane content, after microwaving I transfer broccoli sprouts to a strainer, and allow further myrosinase hydrolization of glucoraphanin and other glucosinolates into sulforaphane and other healthy compounds.

I use the above studies as guides to create broccoli sprout hydrolysis compounds just before eating them. I don’t depend on my metabolism to create sulforaphane, DIM, erucin, and other hydrolysis compounds as did the clinical trial. But then again, those subjects ate super sprouts:

“We used the elicitor methyl jasmonate (MeJA) by priming the seeds as well as by spraying daily over the cotyledons from day 4-7 of germination. We observed that MeJA at concentrations of 250 μmol act as stressor in the plant and enhances the biosynthesis of the phytochemicals glucosinolates.

Compared to control plants without MeJA treatment, the content of compounds as the aliphatic glucosinolate glucoraphanin was enhanced up to a 70% and similar increases were observed with glucoiberin or glucobrassicin. In this way, we improved the content of these health-promoting compounds.”

I don’t have a scale in my kitchen, and don’t have a measured weight of broccoli sprouts consumed daily. It’s probably more than twice the clinical trial’s 30 grams:

  • My most recent broccoli seed purchase was a 5 pound can (2,268 grams). Its volume by the formula height x π x (diameter / 2)2 with 17 cm height and 15 cm diameter is 1,767 cubic centimeters.
  • With 1 tablespoon = 14.79 cc, (14.79 cc / 1,767 cc) x 2,268 grams = 19 grams per serving. Broccoli seed weight of two servings is 19 x 2 = 38 grams a day.
  • The lowest weight gain for 3-day-old broccoli sprouts in Item 2 above was 4.32 times the seed weight. That was in laboratory conditions, though.
  • Let’s guess that 3-day-old broccoli sprouts only gain twice as much weight in my kitchen, 38 g x 2 = 76 grams. A comparable worst-case Estimating daily consumption of broccoli sprout compounds calculation was 75.52 grams.

I’ve referenced our model clinical trial in 15 previous blog posts. They are, in date descending order:

  1. A pair of broccoli sprout studies
  2. Reversal of aging and immunosenescent trends with sulforaphane
  3. A hair color anecdote
  4. Week 7 of Changing to a youthful phenotype with broccoli sprouts
  5. Part 2 of Rejuvenation therapy and sulforaphane
  6. A rejuvenation therapy and sulforaphane
  7. Week 6 of Changing an inflammatory phenotype with broccoli sprouts
  8. Week 3 of Changing an inflammatory phenotype with broccoli sprouts
  9. Broccoli sprouts oppose effects of advanced glycation end products (AGEs)
  10. Reviewing clinical trials of broccoli sprouts and their compounds
  11. Understanding a clinical trial’s broccoli sprout amount
  12. Week 2 of Changing an inflammatory phenotype with broccoli sprouts
  13. Changing an inflammatory phenotype with broccoli sprouts
  14. Growing a broccoli sprouts Victory Garden
  15. How much sulforaphane is suitable for healthy people?