A claim of improved cognitive function

I’ll describe evidence for claiming improved cognitive function in Week 9 of Changing to a youthful phenotype with broccoli sprouts.

I read parts of over a hundred research papers last week. That required substantial concentration to understand them, and stay on topic while learning new items, which started new searches. This wasn’t a new development, it was just to a much greater extent. I also worked forty hours for my job.

The main chain of blog posts began when I relooked at the presentation in Reversal of aging and immunosenescent trends after remembering it included before and after photos per A hair color anecdote. The presentation prompted last week’s most frequent self-question, Why didn’t I see this before?

One possible explanation is that people don’t usually see things outside their conditioned perceptions. (1) Reevaluate findings in another paradigm illustrated this with an example of how different frameworks viewed the same hypothalamus study differently.

I was interested to see what sulforaphane research had in common with the presentation topics, which produced (2) Reversal of aging and immunosenescent trends with sulforaphane. That required gaining a better understanding of PubMed search techniques, which led to (3) A pair of broccoli sprout studies.

Numerous presentation topics resulted in (4) Part 2 of Reversal of aging and immunosenescent trends with sulforaphane. I investigated one of its cited papers in (5) A review of sulforaphane and aging, which required further searches, some of which are still on tabs of my browser.

I was happy to oblige special requests with (6) Tailoring measurements for broccoli sprouts and (7) Uses of the lymphocytes to monocytes ratio.

Could I have done all of what I did last week without changing my internal environment? What exactly are the effects of eating a clinically relevant amount of broccoli sprouts every day for nine weeks?

A plausible explanation is in Upgrade your brain’s switchboard with broccoli sprouts.

A review of sulforaphane and aging

This 2019 Mexican review stated:

“We describe some of the molecular and physical characteristics of SFN, its mechanisms of action, and the effects that SFN treatment induces in order to discuss its relevance as a ‘miraculous’ drug to prevent aging and neurodegeneration. SFN has been shown to modulate several cellular pathways in order to activate diverse protective responses, which might allow avoiding cancer and neurodegeneration as well as improving cellular lifespan and health span.

NF-κB is in charge of inflammatory response regulation. Under basal conditions, NF-κB is sequestrated into the cytosol by IκB, but when pro-inflammatory ligands bind to its receptors, the IKK protein family phosphorylates IκB to degrade it via proteasome, so NF-κB is able to translocate into the nucleus and transcript several inflammatory mediators. Sulforaphane is capable to inhibit IκB phosphorylation and NF-κB nuclear translocation.

SFN upregulated Nrf2 expression by reducing DNA demethylation levels of the Nrf2 promoter. In another model using the triple-transgenic mouse model of Alzheimer’s disease (3 × Tg-AD), the use of SFN regulates the expression of the Brain-derived neurotrophic factor (BDNF) via HDAC inhibition, thus increasing H3 and H4 acetylation on the BDNF promoter. Enhancing BDNF expression as an effect of SFN treatment increased the neuronal content of several synaptic molecules like MAP 2, synaptophysin, and PSD-95 in primary cortical neurons of 3 × Tg-AD.”

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6885086/ “Sulforaphane – role in aging and neurodegeneration”

I came across this review while searching PubMed for sulforaphane commonalities with presentation topics in Part 2 of Reversal of aging and immunosenescent trends with sulforaphane. The review outlined some aging aspects and presented relevant sulforaphane studies. Others such as eye and muscle decline weren’t addressed.

Since sulforaphane’s “a ‘miraculous’ drug” in the Abstract, I expected but didn’t see corresponding excitement in the review body. Just phrases like “it is known” and non-specific “more research is needed.”

Other papers published after this review were found by a PubMed “sulforaphane signal aging” search:

A pair of broccoli sprout studies

This 2015 Oregon human study found:

  • “Plasma and urinary levels of total SFN [sulforaphane] metabolites were ~3–5 times higher in sprout consumers compared to BSE [broccoli sprout extract] consumers.
  • In sprout consumers, plasma concentrations were 2.4-fold higher after consuming the second dose than after the first dose.
  • Calculated SFN bioavailability from broccoli sprouts exceeded 100%.

a Following consumption of a single 200-µmol SFN dose. b Cumulative excretion of SFN metabolites from baseline collection through the 48-hr study period. c Bioavailability F calculated based on total micromoles excreted in urine. Cmax, maximum concentration observed; AUC, area under the curve; h, hour; L, liter; t1/2, half-life; Tmax, time at Cmax. Values represent mean ± SD, n = 10.”

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4394840/ “Absorption and chemopreventive targets of sulforaphane in humans following consumption of broccoli sprouts or a myrosinase-treated broccoli sprout extract”

Another way to state findings:

  1. Broccoli sprouts are better than supplements.
  2. Eating sprouts twice a day is better than eating them once a day.

No explanation was given for sprout weight variability, although one was needed, because 127.6 g / 2 = 63.8 g, not 46.8 g:

“In the divided-dose phase (two weeks later), subjects (fasting) consumed half the original dose (100 μmol SFN equivalents) at 8 AM from sprouts or the BSE and the other half (not fasting) 12 h later.”

A “SFN potential” process demonstrated sulforaphane amount equivalencies, but didn’t explain non-intuitive sprout weight measurements. Was it too difficult to control sprout variability? The difficulties were instead pushed onto other researchers trying to replicate the study, and consumers looking for practical guidance.

Regardless, I adjusted my practices to twice daily start a new broccoli sprout batch with one tablespoon of seeds rather than once a day with two tablespoons. I eat them with breakfast and dinner rather than at lunch.

I microwave 3-day-old sprouts immersed in 100 ml distilled water on full 1000W power for 40 seconds to achieve 58°C. I immediately put them into a strainer to allow further myrosinase hydrolization of glucoraphanin and other glucosinolates into sulforaphane and other healthy compounds.

I’d overlooked the above study until I saw it referenced in its successor 2018 human study Untargeted metabolomic screen reveals changes in human plasma metabolite profiles following consumption of fresh broccoli sprouts and cited it in Reversal of aging and immunosenescent trends with sulforaphane for its DHEA findings. The clinical trial treatments included:

“Both dehydroepiandrosterone (DHEA) and metformin in an attempt to limit the “diabetogenic” effect of GH [growth hormone]. DHEA has many effects, in both men and women, that oppose deleterious effects of normal aging.”

A PubMed search on DHEA found Impact of Dehydroepianrosterone (DHEA) Supplementation on Serum Levels of Insulin-Like Growth Factor 1 (IGF-1): A Dose-Response Meta-Analysis of Randomized Controlled Trials which confirmed the clinical trial’s DHEA dose would increase IGF-1.

This study observed a significant decrease in DHEA after eating broccoli sprouts, but didn’t provide a plausible explanation for this finding, or cite relevant studies. Ten other significant decreases were related to antioxidants and fatty acids.

It isn’t clear that I needed to take DHEA anyway, since the clinical trial’s purpose for DHEA treatment was to oppose effects of growth hormone, which I’m not taking. But I’m getting good results, so I’ll just keep doing what I’ve been doing for a limited time.

The study said:

“While this study focuses largely on the potential effects of SFN, broccoli sprouts contain many other bioactive components that could be responsible for our observations as well as additional health benefits.”

Our model clinical trial Effects of long-term consumption of broccoli sprouts on inflammatory markers in overweight subjects said much the same:

“The anti-inflammatory effects observed with broccoli sprouts intake are likely due to the combined effects of all the hydrolysis products of glucosinolates.”

The 3-day-old broccoli sprouts have the optimal yields study said:

Although germination reduces SF [sulforaphane] yield to some extent, it is beneficial to the formation and accumulation of total phenol and flavonoids, ensuring the health properties of sprouts.”

Combining the pair of Oregon studies’ findings:

  1. Broccoli sprouts are better than supplements.
  2. Eating sprouts twice a day is better than eating them once a day.
  3. When in doubt, refer back to Item 1.


Do broccoli sprouts treat migraines?

While rereading a review in Eat broccoli sprouts today, it occurred to me that I haven’t needed to take migraine medicine during the 9 weeks I’ve been eating broccoli sprouts every day. Since 14 weeks of lockdown overlap this period, it’s also possible that I’ve avoided triggering conditions. I look at brightly-lit screens all day, but don’t have cold air blowing on my head that’s the other half of my most common triggering condition.

I started having intermittent ~monthly episodes about ten years ago. I wouldn’t take sumatriptan unless I have a half-day-long headache that doesn’t respond to acetaminophen. It stops a headache from turning into a 3-day-long migraine.

I went over to PubMed and did a “sulforaphane migraine” search, which turned up exactly 1 (!!) result. A 2016 Chinese rodent study Activation of the nuclear factor E2-related factor 2/anitioxidant response element alleviates the nitroglycerin-induced hyperalgesia in rats found:

“Activation of the Nrf2/ARE pathway inhibited the activation of TGVS [trigeminovascular system] and prevented the induction of hyperalgesia. Sulforaphane might therefore be an effective agent for hyperalgesia.”

Plausible conclusion. Nitroglycerin definitely jolts a monster headache. 5 mg sulforaphane / kg body weight was a large dose as well.

Two of the eleven papers citing this study were:

There wouldn’t be any potential payoff for a company to be interested in studying a sulforaphane-migraine connection. What sponsor would be interested enough to double the number of studies in this area?

A hair color anecdote

Will you excuse a poorly-evidenced observation that’s a positive development I left out of Week 8 of Changing to a youthful phenotype with broccoli sprouts?

I got a haircut last weekend after waiting for Governor Klan Robes Blackface to not arrest barbershop and hair salon owners for the crime of earning a living. A thirty-something tattooed barber wearing a face mask and face shield said my last haircut had been on February 1, 2020, so it had been 14 weeks. She used a #4 clipper to cut everything to about 1/2 inch.

I’d eaten broccoli sprouts every day for 7 weeks at that point. Post-haircut visible hair was all from that period, probably since Week 3, which was also when broccoli sprouts’ effects on inflammation became noticeable.

One evening as I brushed my teeth, I noticed overall hair appearance was mainly dark brown again, an unexpected phenomenon. Maybe white hair will show up as it gets longer?

Feedback on hair color from a back-of-the-head picture was mixed, ranging from “Yes. Definitely!” to Unsupported non-evidence since before and after pictures weren’t taken under the same lighting conditions. Even if validated, other factors could be in play, such as working from home without the stress of going into work.

While eating my usual steel cut oats for breakfast this morning, I remembered a super informative presentation by the lead researcher of clinical trial Reversal of aging and immunosenescent trends. I rewatched it, pausing after two minutes to reabsorb when he said:

“There’s a collapse that takes place somewhere between the ages of sixty to eighty in which you lose 98% of your ability to recognize foreign antigens.”

You will have forgotten why I drew your attention to this super interesting presentation by the 21:25 mark. But pause for the “Hair Rejuvenation?” slide with before and after photos:

“A couple of guys came to us and said they seemed to notice that their hair was growing in darker again. It’s an anecdote. It didn’t apply to most of the guys. But it’s a sign that maybe something interesting is going on.”

That’s followed by epigenetic clock findings using four different clocks. Note that no significant effects on biological age were found until the trial’s 9-month point, and those weren’t as strong as improvements by 12 months.

Improvements accelerated between 9 and 12 months, and at 12 months, subjects had increased their life expectancies by 2 years. The GrimAge clock showed the subjects’ predicted lifespan and health span was unchanged 6 months after the trial ended.

I started and have continued four lifestyle “interventions” since last summer:

  1. In July I dramatically reduced 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 this trial’s non-prescription daily treatments of Vitamin D, zinc, and DHEA.
  3. Also in September, I started non-prescription intermittent quercetin treatments of Preliminary findings from a senolytics clinical trial.
  4. Eight weeks ago I started eating broccoli sprouts every day per clinical trial Effects of long-term consumption of broccoli sprouts on inflammatory markers in overweight subjects.

In a month or so I should be able to say whether or not my hair really is growing in darker. One way to find out which “intervention” had the largest effect may be to stop one or more of them. That might happen anyway because:

  1. Consistently eating AGE-less food is boring.
  2. I’m leery of taking more than RDAs.
  3. Ehh.
  4. I still sadly hope against reality that we’re past the Madness of Crowds phase and can accelerate the “recover their senses slowly, one by one” phase. It would be harder to take care of my broccoli sprout farm if I have to go into work every day.

Or maybe An environmental signaling paradigm of aging is correct, and at a certain point, clocks are reset and none of these “interventions” will be needed? What do you think?


Lab analyses of broccoli sprout compounds

To follow up Estimating daily consumption of broccoli sprout compounds, the main point of which was: how can a person guide their actions with evidence when a broccoli cultivated variety’s beneficial characteristics aren’t known?

I focus on broccoli sprout lab analyses of 3 cultivars made in a 2017 German study Isothiocyanates, Nitriles, and Epithionitriles From Glucosinolates Are Affected by Genotype and Developmental Stage in Brassica oleracea Varieties. Here are rounded numbers from the supplementary material:

1. What happened in 2015 to lessen all broccoli cultivars’ sprout compound values by 2015 / 2014 percentages?

I read and reread, and didn’t come up with a satisfactory answer. Maybe it’s a language difference, but this is an insufficient explanation:

“Decreased myrosinase activity probably was the reason for the low levels of hydrolysis products formed in broccoli cv. Sirtaki in 2015 compared to 2014, while 70% [67%] of the GLS concentration of 2014 was present.”

What caused “decreased myrosinase activity?” And why would that be explanatory for just one cultivar when two others also experienced “low levels of hydrolysis products?”

Greenhouse-grown broccoli sprouts clearly varied between batches. Temperatures and sun radiation were tossed in but not proven as explanatory variables.

2. Could a person infer from one cultivar’s 2014 lab report how much sulforaphane they would consume from that cultivar in 2015? No.

3. Without knowing the broccoli sprouts’ cultivar, could a person infer from a glucoraphanin amount how much sulforaphane they would consume?

No. The 2014 Iron Man 5.2 μmol per g glucoraphanin wasn’t that different from the 2015 Sirtaki 5.4 amount. But Iron Man vs. Sirtaki differences of 0.3 vs. 0.6 μmol per g sulforaphane amounts and 5% vs. 11% hydrolyzed showed Sirtaki cultivar’s unrounded numbers had double or more those of Iron Man.

4. Could a person infer from a mature broccoli glucoraphanin amount anything about its broccoli sprout glucoraphanin amount, or vice versa?

No. The Sirtaki 8.0 μmol per g sprout glucoraphanin amount was the highest cultivar, but its fully developed head was lowest at 0.27.

5. Could differences be explained by the broccoli sprout batches’ one-day age difference?

6. Did both years use the same seed lot? If so, could a one year aging of seeds explain differences?

Here’s a graphic of the 2014 batch major broccoli sprout compounds. Glucoraphanin is red 4MSOB in A, and sulforaphane is red 4MSOB-ITC in C:

This study demonstrated that are many unknowns even with lab analyses. It would seem that not much could be accurately inferred past knowing that broccoli sprouts of one cultivar produced more sulforaphane than another.

Would similar lab reports help current purchase decisions? I haven’t come across a broccoli seed bulk supplier who had lab reports for products they’re selling. When pressed on Amazon they at best say Calabrese, which has described hundreds of cultivars. Such as Iron Man and Marathon, which are also named Calabrese Iron Man F1 and Calabrese Marathon F1.

Obtaining information for each batch would probably cost them a lot of money, which would be passed on to their customers. Lab reports for a different batch than the broccoli seed offered may have limited utility to consumers.

It would assist broccoli seed consumers to at least know the cultivar. This study’s genotypes had a greater effect than did environmental influences.

See follow up Tailoring measurements for broccoli sprouts.

Enhancing sulforaphane content

This 2020 Chinese study experimented with enhancing sulforaphane content of broccoli florets in a range of conditions:

“For direct water blanching at 60°C, the sulforaphane yield increased with treatment time from 1698.0 ± 121.9 μmol per kg DW (0 min) to 2833.3 ± 118.6 μmol per kg DW (1 min) and then steadily decreased to the lowest value of 2345.8 ± 57.7 μmol per kg DW for 5 min.”

The sulforaphane yield was 503.7 ± 23.8 μmol per kg DW of broccoli after 5 min thermal treatment at 65 °C, which was even lower than the value obtained for raw broccoli. The reason could be the leaching of glucoraphanin into the blanching water coupled with partial inactivation of myrosinase resulting in low yield of sulforaphane.

For direct water blanching, the best treatment temperature for maximizing sulforaphane yield was 60 °C, which is similar to the best treatment temperature for maximizing sulforaphane yield reported previously.

Sulforaphane yield depends on the relative activity of myrosinase and ESP in the broccoli matrix and 3 min treatment at 65 °C during in-pack processing in this study was found to be the best condition that favours conversion into sulforaphane instead of sulforaphane nitrile. This indicates that the condition favours the inactivation of ESP to a larger extent while maintaining sufficient myrosinase activity resulting in optimal conversion into sulforaphane.

Under this condition, it seems that all of the extractable glucoraphanin is converted to sulforaphane assuming 1 to 1 conversion, since the glucoraphanin content of the broccoli samples were determined to be 3141.2 μmol per kg DW whereas the sulforaphane yield was 3983 μmol per kg DW. The slightly higher sulforaphane yield than would be predicted from the level of glucoraphanin in raw broccoli requires further investigation.”

https://pubs.rsc.org/en/content/articlehtml/2020/fo/c9fo02089f “Mild heat combined with lactic acid fermentation: a novel approach for enhancing sulforaphane yield in broccoli puree”

1. The study presented evidence for kitchen practices:

  • Per the above graphic’s point a, I’ve changed to let broccoli sprout heating continue for 1 minute after microwaving to achieve 60°C. This allows further myrosinase hydrolization of glucoraphanin into sulforaphane. My practice had been to immediately cool them down, which was suboptimal point c on the 60°C line. I still transfer the broccoli sprouts to a strainer immediately after microwaving.
  • The 60°C (140°F) cliff finding of Microwave broccoli to increase sulforaphane levels was confirmed.

2. I didn’t view this study’s in-pack or lactic acid bacteria fermentation findings as having practical kitchen use. Maybe it’s a cultural difference?

3. Poor performance at 65°C after 5 minutes was partially attributed to “leaching of glucoraphanin into the blanching water.” But poor 65°C performance was evident at the 1 minute point compared with good 60°C performance.

“Partial inactivation of myrosinase” at 65°C was more likely to be the dominant factor.

4. Regarding:

“The slightly higher sulforaphane yield than would be predicted from the level of glucoraphanin in raw broccoli requires further investigation.”

The microwaving study author was on a productive investigation track with:

“Microwave irradiation might help to release more conjugated forms of glucosinolates and then get hydrolyzed by released myrosinase.”

That track developed in part from finding that broccoli florets microwaved on full power to 60°C increased glucoraphanin past control (raw) levels:

“The control GLR amount was 2.18 µmol/g DW, while the HL60 GLR amount was 2.78 µmol/g DW.”

Not to mention the coincident 1,114% increase in sulforaphane content of ordinary broccoli purchased at a grocery store!

I arrived at this study through it being referenced in the enjoyable 2020 Spanish review Functional Ingredients From Brassicaceae Species: Overview and Perspectives. The reviewers noted that this study’s 2019 predecessor Fermentation for enhancing the bioconversion of glucoraphanin into sulforaphane and improve the functional attributes of broccoli puree (not freely available) found:

“Preferential formation of SFN-nitrile (less potential as inducer of phase II detoxification enzymes than SFN) instead of SFN.”