Stress

Eat broccoli sprouts for DIM

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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 30 grams of broccoli super 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 metabolizing more DIM.

That Day 35 data point didn’t have an asterisk next to it to indicate a statistically significant decrease. But the overweight postmenopausal women group’s next Day 50 significant “steady decrease” finding supported an interpretation that eating broccoli sprouts supplied them 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 super sprout indolic compounds were as follows:

indolic glucosinolates

Assuming that only glucobrassicin is a precursor to DIM, subjects’ DIM bioavailability can be calculated as μmol DIM / 21.61 μmol. For example, overweight postmenopausal women Day 35 average of 0.5544 μmol DIM that ranged from 0.1771 to 0.8034 μmol DIM represented an average 2.57% DIM bioavailability with a range of 0.82% to 3.72% DIM bioavailability.

See Part 2 for DIM follow-up.

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

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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.

IL-6 2020

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.” [Zinc and broccoli sprouts – a winning combination]
  • “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 estimated 52 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 = 21.4 g x 30 days = .642 kg or 1.42 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

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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. I eat 131 grams daily as calculated in Estimating daily consumption of broccoli sprout compounds.
  3. 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.
  4. I immerse 3-day-old broccoli sprouts in 100 ml distilled water, then microwave them on 1000W full power for 35 seconds to ≤ 60°C (140°F) per Microwave broccoli to increase sulforaphane levels.
  5. 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, indole-3-carbinol, 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’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?

Week 9 of Changing to a youthful phenotype with broccoli sprouts

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To follow up Week 8 of Changing to a youthful phenotype with broccoli sprouts:

1. This week has really been different.

A. Physically, on Friday Eve I worked out per my usual upper-body-workout-every four-days routine. I felt strong, and on one exercise I increased the weight by 33%. No problem doing the same number of reps and sets! Keeping good form was challenging.

Per Week 7, I eight-count each concentric rep slowly, then perform each eccentric rep to the same count, with a goal to reach muscle exhaustion during each set. Then pause and do another set.

What changed? Could I have done all this before?

No. I’d tried, making baby steps with increasing weight and keeping good form. But now I can, and I’ll do it again, along with other physical challenges.

B. Seven blog posts this week show improved cognitive function. Is A claim of improved cognitive function sufficient evidence?

Awakening was how it felt. Waking up to what I didn’t see before.

C. This 35th blog post for May comes after 30 posts in April. It wasn’t my goal to do one a day. It’s my goal to Surface Your Real Self. Did a few of them help?

I hope to do other things with my life in June. But the fact remains that humans are herd animals. We “think in herds, go mad in herds, while they [we] only recover their [our] senses slowly, one by one.” We’ll stay in the Madness of Crowds phase until enough people refuse to be propagandized.

2. As a result of reading A pair of broccoli sprout studies, I changed practices to start batches with one tablespoon of broccoli seeds twice a day so I could consume broccoli sprouts twice daily. Right now it’s a PITA task that requires optimization.

The two studies’ findings were:

  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.

3. I reordered broccoli seeds and will receive them next week. In the meantime, I introduced yet another unknown by consuming sprouts that came from a different vendor:

These seeds are smaller. Hundreds of seeds and seed coats annoyingly pass through my strainer, which didn’t happen with larger seeds. 3-day-old sprout sizes are smaller, and they smell and taste different.

This vendor put “seed” four times on their label. The other vendor didn’t bother to put “seed” even once on their broccoli seed package label.

Like other vendors, they prefer buzzword marketing with “microgreen” and “sprouting” rather than provide useful consumer information such as number of seeds and broccoli variety characteristics. Will people buy “Broccoli Sprouting Seeds” but won’t buy Broccoli Seeds? Do people say “Cool beans!” anymore?

My reorder states there are ~720,000 broccoli seeds in that 5 lb. package. I’ll update with its volume after it arrives.

See Week 10 of Changing to a youthful phenotype with broccoli sprouts for follow ups.

Uses of the lymphocytes-to-monocytes ratio

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To follow up a presentation topic of Part 2 of Reversal of aging and immunosenescent trends with sulforaphane, here are a few papers no earlier than 2015 that address the ratio of lymphocytes to monocytes (LMR), or its reciprocal MLR. Because inquiring minds want to know. 🙂

  1. Monocyte heterogeneity and functions in cancer

    “The ratio of lymphocytes to monocytes has emerged as a prognostic factor, including for B cell lymphoma, colorectal cancer, lung cancer, and ovarian cancer. For example, in patients with stage III colon cancer, a higher lymphocyte to monocyte ratio was associated with increased time to recurrence and overall survival.”

  2. Distinct Transcriptional and Anti-Mycobacterial Profiles of Peripheral Blood Monocytes Dependent on the Ratio of Monocytes: Lymphocytes

    “Our observation of monocyte functional and transcriptional differences dependent on the ML ratio (but on neither constituent alone) suggests that qualitative differences in monocytes are better reflected by the ML ratio than by monocyte counts alone, potentially explaining epidemiologic associations of the ratio. The ML ratio was associated with mycobacterial growth in vitro (β = 2.23, SE 0.91, p = 0.02). The significant enrichment of interferon signalling we found supports a common role for type I and II interferons in altering the ML ratio and monocyte function sufficiently to explain altered disease course, consistent with the central role of interferons in mycobacterial and inflammatory diseases. In humans, myeloid-biased HSC accumulate with age and explain the relative increase in myeloid cells in blood with age. Therefore changes in ML ratio in blood are likely a marker of changes in the frequency of lineage-biased HSC.”

  3. Monocyte–lymphocyte ratio is a valuable predictor for diabetic nephropathy in patients with type 2 diabetes

    “T2D patients without diabetic-related complications had higher MLR than control patients. MLR was significantly higher in DN patients than in T2D patients without diabetic-related complications.”

  4. Monocyte lymphocyte ratio predicts the new-onset of chronic kidney disease: A cohort study

    “Increased baseline MLR is strongly associated with the risk of new-onset CKD in people with normal or near-normal kidney function at baseline. Inflammatory markers such as interleukin are difficult to be measured by primary medical care. Therefore, search for simpler inflammatory markers to predict the risk of CKD. MLR represent[s] the state of balance between inflammatory activators and inflammatory regulators. The higher the ratio, the greater the imbalance, the more severe the inflammatory response and the stronger the immune suppression. In addition to increasing the risk of new-onset CKD, our study found MLR was positively related to inflammatory factors, such as leukocytes, neutrophils, NLR, PLR and platelet distribution width. In addition, MLR was positively correlated with age, blood pressure and BMI. However, there was no significant correlation between MLR and fasting plasma glucose in non-diabetic participants. A total of 11280 participants (6592 male and 4688 female) were enrolled in this longitudinal study.”

A LMR of 5 and a MLR of 0.2 are easy-to-measure heuristics, adequate for screening people. These ratios can be used along with many other measurements as starting points to investigate underlying causes.

Item 1 described how LMR also has prognostic value for cancers. The other studies used MLR as a biomarker for the future course of inflammatory diseases per:

“The higher the ratio, the greater the imbalance, the more severe the inflammatory response and the stronger the immune suppression.”

I’d seen the below presentation graphic several times since September 2019. My reaction was “Oh, that’s interesting” each time.

On Friday I understood it: This was what resetting your internal environment looked like.

Did my paradigm change? Yes, among other things, and all of that allowed me to see.

An environmental signaling paradigm of aging provided evidence up through 2015 for its hypothesis and framework. Its treatments’ capabilities to “reset to different age-phenotypes“ will be tested as the 2020 study underlying A rejuvenation therapy and sulforaphane is tested.

Caution is warranted before getting carried away with ratio analyses of a 9-subject pilot study. Are hormone ratios useful in explaining health? Behavior? Neurobiology? Anything? recommended:

“Analysis of the individual variables offers more information and a more accurate picture of the underlying relationships.

Ratios should either be analyzed with non-parametric techniques, or be log-transformed before parametric statistical methods are applied.”

There was monocyte but not lymphocyte data in the clinical trial’s supplementary material.

A review of sulforaphane and aging

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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:

Part 2 of Reversal of aging and immunosenescent trends with sulforaphane

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Reversal of aging and immunosenescent trends with sulforaphane covered only the first 13 minutes of a super informative presentation by the lead researcher of clinical trial Reversal of aging and immunosenescent trends.  Commonalities with sulforaphane research were found by PubMed searches of sulforaphane and each presentation topic, and used a 1/1/2015 publication date cutoff.

Continuing presentation topics from the 13:40 mark:

Cancer

Lymphocyte/monocyte ratio

CD38 monocytes

  • NQO1-induced activation of AMPK contributes to cancer cell death by oxygen-glucose deprivation

    “NQO1 plays a key role in AMPK-induced cancer cell death in OGD through the CD38/cADPR/RyR/Ca2+/CaMKII signaling pathway. Expression of NQO1 is elevated by hypoxia/reoxygenation or inflammatory stresses through nuclear accumulation of the NQO1 transcription factor, Nrf2 (NFE2-related factor 2). Activation of the cytoprotective Nrf2 antioxidant pathway by sulforaphane protects immature neurons and astrocytes from death caused by exposure to combined hypoxia and glucose deprivation.”

Thymus – no recent sulforaphane studies

Renal function

  • Rapid and Sustainable Detoxication of Airborne Pollutants by Broccoli Sprout Beverage: Results of a Randomized Clinical Trial in China

    “Rapid and sustained, statistically significant increases in levels of excretion of glutathione-derived conjugates of benzene (61%), acrolein (23%), but not crotonaldehyde were found in those receiving broccoli sprout beverage compared with placebo. Excretion of benzene-derived mercapturic acid was higher in participants who were GSTT1-positive compared to the null genotype, irrespective of study arm assignment. Measures of sulforaphane metabolites in urine indicated that bioavailability did not decline over the 12-week daily dosing period. Intervention with broccoli sprouts enhances detoxication of some airborne pollutants, and may provide a frugal means to attenuate their associated long-term health risks.”

Hair rejuvenation

Epigenetic clocks – There are no sulforaphane studies that use epigenetic clocks, although broccoli compounds have epigenetic effects on aging, as reviewed in 2019:

  • Sulforaphane – role in 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.”

Both biomarker (Lymphocyte / monocyte ratio) and epigenetic clock (GrimAge) measurements done 6 months after the clinical trial ended suggested trial subjects’ aging phenotypes had been reset:

An environmental signaling paradigm of aging explained:

“Apart from being slowed down or sped up, the body clock can also be reset. Organisms, organs, and their cells can be reset to different age-phenotypes depending on their environment.

This is not so much a principle as an application of principle that the environment determines age-phenotype.”

There wouldn’t be a potential payoff for a company to study any broccoli compound / aging connections. People can achieve clinically relevant, daily doses of broccoli sprouts for < $500 a year.

What sponsor would be interested enough to put sulforaphane research on the clock?

Presentation topics are continued in Uses of the lymphocytes to monocytes ratio and A review of sulforaphane and aging. This pilot trial’s follow-on clinical trial was updated in The next phase of reversing aging and immunosenescent trends.

Reversal of aging and immunosenescent trends with sulforaphane

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Sulforaphane research findings have commonalities with a super informative presentation by the lead researcher of clinical trial Reversal of aging and immunosenescent trends. I did a PubMed search of sulforaphane and each presentation topic, and used a 1/1/2015 publication date cutoff.

Presentation topics through the first 13 minutes were:

Thymus – no recent sulforaphane studies

Treatments

PSA

C-reactive protein and IL-6

Bone marrow fat – no recent studies

T cells

PD-1 / PD-L1

Treatment cost

I estimate the annual cost of the non-prescription treatments of the clinical trial to be $100. The estimated annual cost of eating broccoli sprouts every day is < $500 for the broccoli seeds.

broccoli seed label

The above image isn’t an endorsement although it’s what I’ve used. It’s buzzword marketing to put “sprouts” and “sulforaphane” but not “seeds” on the label of a broccoli seeds package. For another thing, broccoli sprouts don’t “abound with phytochemical sulforaphane.”

Repeating a point from Estimating daily consumption of broccoli sprout compounds, broccoli seeds and sprouts contain little or no sulforaphane. They have glucoraphanin and myrosinase enzyme which are structurally separated. Disturbing their cells mixes the two, and the enzyme hydrolyzes glucoraphanin and other glucosinolates into sulforaphane and other healthy compounds.

Continue presentation topic commonalities with sulforaphane research at A pair of broccoli sprout studies and Part 2 of Reversal of aging and immunosenescent trends with sulforaphane.

Reevaluate findings in another paradigm

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It’s challenging for people to change their framework when their paychecks or mental state or reputations depend on it not changing.

I’ll use The hypothalamus and aging as an example. This review was alright for partial fact-finding up through 2018. Its facts were limited, however, to what fit into the reviewers’ paradigm.

The 2015 An environmental signaling paradigm of aging provided examples of findings that weren’t considered in this 2018 review. It also presented a framework that better incorporated what was known in 2015.

Here’s how they viewed the same 2013 study, Hypothalamic programming of systemic ageing involving IKK-β, NF-κB and GnRH (not freely available).

Paradigm: “The hypothalamus is hypothesized to be a primary regulator of the process of aging of the entire body.”

Study assessment:

“Age-associated inflammation increase is mediated by IκB kinase-β (IKK-β) and nuclear factor κB (NF-κB) in microglia and, subsequently, nearby neurons through microglia–neuron interaction in the mediobasal hypothalamus. Apparently, blocking hypothalamic or brain IKK-β or NF-κB activation causes delayed aging phenotype and improved lifespan.

Aging correlates with a decline in hypothalamic gonadotropin-releasing hormone (GnRH) expression in mice. Mechanistically, activated IKK-β and NF-κB significantly down-regulates GnRH transcription. GnRH therapy through either hypothalamic third ventricularor or subcutaneous injection leads to a significant recovery of neurogenesis in the hypothalamus and hippocampus, and a noticeable improvement of age-related phenotype in skin thickness, bone density, and muscle strength when applied in middle-aged mice.”

Paradigm: Environmental signaling model of aging

Study assessment:

“A link between inflammation and aging is the finding that inflammatory and stress responses activate NF-κB in the hypothalamus and induce a signaling pathway that reduces production of GnRH by neurons. GnRH decline contributes to aging-related changes such as bone fragility, muscle weakness, skin atrophy, and reduced neurogenesis. Consistent with this, GnRH treatment prevents aging-impaired neurogenesis, and decelerates aging in mice.

Zhang et al. report that there is an age-associated activation of NF-κB and IKK-β. Loss of sirtuins may contribute both to inflammation and other aspects of aging. But this explanation, also given by Zhang et al., merely moves the question to why there is a loss of sirtuins.

The case is particularly interesting when we realize that the aging phenotype can only be maintained by continuous activation of NF-κB – a product of which is production of TNF-α.

Reciprocally, when TNF-α is secreted into the inter-cellular milieu, it causes activation of NF-κB. In their study, Zhang et al. noted that activation of NF-κB began in microglia (the immune system component cells found in the brain), which secreted TNF-α, resulting in a positive feedback loop that eventually encompassed the entire central hypothalamus.

The net result of this is a diminution in production of gonadotropin-releasing factor which accounted for a shorter lifespan. Provision of GnRH eliminated that effect, while either preventing NF-κB activation (or that of the IKK-β upstream activator) or by providing gonadotropin-releasing factor directly into the brain, or peripherally, extending lifespan by about 20%.

In spite of the claim of Zhang et al. that the hypothalamus is the regulator of lifespan in mice, their experiments show that only some aspects of lifespan are controlled by the hypothalamus, as preventing NF-κB activation in this organ did not stop aging and death. Similar increased NF-κB activation with age has been seen in other tissues as well, and said to account for dysfunction in aging adrenal glands.

It was demonstrated that increased aging occurred as a result of lack of gonadotropin-releasing hormone, and that increased lifespan resulted from its provision during aging. In this manner:

  1. Aging of hypothalamic microglia leads to
  2. Aging of the hypothalamus, which leads to
  3. Aging elsewhere in the body.

So here we have a multi-level interaction:

  1. Activation of NF-κB leads to
  2. Cellular aging, leading to
  3. Diminished production of GnRH, which then
  4. Acts (through cells with a receptor for it, or indirectly as a result of changes to GnRH-receptor-possessing cells) to decrease lifespan.

So the age state of hypothalamic cells, at least with respect to NF-κB activation, is communicated to other cells via the reduced output of GnRH.”

Not using the same frameworks, are they?

In 2015, this researcher told the world what could be done to dramatically change the entire aging research area. He and other researchers did so recently as curated in Part 3 of Rejuvenation therapy and sulforaphane which addressed hypothalamus rejuvenation.

A hair color anecdote

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

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 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?

Week 8 of Changing to a youthful phenotype with broccoli sprouts

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

To follow up Week 7 of Changing to a youthful phenotype with broccoli sprouts:

1. I changed practices per Enhancing sulforaphane content. After microwaving to achieve 60°C, I now transfer broccoli sprouts to a strainer, and allow further myrosinase hydrolization of glucoraphanin and other glucosinolates into sulforaphane and other healthy compounds. I previously cooled them immediately.

They taste better, and I stopped putting mustard in them to make them more palatable. What does letting 3-day-old broccoli sprouts cool down by themselves to increase sulforaphane do that makes them more agreeable?

Despite improving yields two weeks ago, 3-day-old broccoli sprouts started from two tablespoons of broccoli seeds still fit into a Corning Ware 16 fl. oz. / 473 ml container:

2. I made a worst-case estimate in Estimating daily consumption of broccoli sprout compounds of 52 mg sulforaphane with microwaving 3-day-old broccoli sprouts. This exceeds:

“The daily SFN [sulforaphane] dose found to achieve beneficial outcomes in most of the available clinical trials is around 20-40 mg.”

The post’s point was: how can a person guide their actions with evidence when a broccoli cultivated variety’s beneficial characteristics aren’t known? I’ll repeat a sulforaphane yields graphic from the 3-day-old broccoli sprouts have the optimal yields study for examples of unknowns:

A. If sulforaphane content was a consumer’s overriding concern. the above evidence suggests that it would be better to always eat the seeds of an unknown cultivar. A tablespoon seems like a good choice, but be sure to chew the broccoli seeds thoroughly (try for five minutes) to release myrosinase and glucoraphanin.

The first minute goes alright. Sometime after that, your mouth and the back of your throat starts to burn. That will be a reminder of an evolved function that protects plants from predators.

I haven’t successfully swallowed a mouthful of thoroughly chewed broccoli seeds without also eating something else or drinking more than just water. That might not go along with your plan for a snack or eating before bedtime.

B. The study recommended consuming 3-day-old sprouts because:

“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.”

Fine, but if your unknown cultivar’s sulforaphane characteristics look like the third cultivar’s 3-day-old sprouts, you’ll have a 53% reduction in the sulforaphane weight. Should you take a 1-in-6 chance with Day 5 sprouts? Or stick with Day 3, guessing that they may still yield more sulforaphane than 3 of the 5 other cultivars’ Day 3 broccoli sprouts?

C. What if you can’t stomach the appearance of 3-day-old broccoli sprouts per the above photo, and you prefer microgreens? Should you wait until Day 7, and take a 1-in-6 chance that your unknown cultivar’s characteristics are like the highest Day 7 of the fourth cultivar? When you roll the die, does it come up 4?

Broccoli seed bulk suppliers aren’t providing evidence for their products and educating customers. Their marketing strategy depends more on buzzwords and price.

3. I compared lab reports of 3 broccoli sprouts’ cultivars in Lab analyses of broccoli sprout compounds to see if they helped rationally deal with these unknowns. It turned out that not much could be accurately inferred from lab reports, past knowing that broccoli sprouts of one cultivar produced more sulforaphane than another.

I haven’t found studies of cultivar characteristics for items I could actually purchase in bulk. I contacted five small US and Canadian suppliers to ask “Do you sell broccoli seeds that have lab evidence of the cultivar’s sulforaphane content?” Two said no so far. I contacted another supplier for the home garden business who has two dozen cultivars listed for sale and asked them the same question.

None of the broccoli seed bulk suppliers specified the cultivar on their offering. When pressed on Amazon they at best said Calabrese, which has described hundreds of cultivars. Such as two in this study, Iron Man and Marathon, which are also named Calabrese Iron Man F1 and Calabrese Marathon F1.

4. I’ve had only sporadic inflammation, and I’m tempted to write anecdotes of positive things. But self-reports are better evidence for emotions than for other internal events.

See Week 9 of Changing to a youthful phenotype with broccoli sprouts for follow ups.

Enhancing sulforaphane content

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

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

“For direct water blanching at 60°C, 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 of broccoli after 5 min thermal treatment at 65 °C was even lower than the value obtained for raw broccoli. The reason could be leaching of glucoraphanin into 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.

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 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 extractable glucoraphanin is converted to sulforaphane assuming 1 to 1 conversion, since glucoraphanin content of 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 up to but no more than 60°C (140°F). 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 broccoli sprouts to a strainer immediately after microwaving.
  • The 60°C cliff finding of Microwave broccoli to increase sulforaphane levels was confirmed by this study’s 65°C direct blanching finding.

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. Those 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.”

Estimating daily consumption of broccoli sprout compounds

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

Strikethroughs are mainly from Week 18 and Week 19 of Changing to a youthful phenotype with broccoli sprouts measurements.

Rainy day thought for the 13th week of lockdown: What are methods of estimating the minimum contents of broccoli sprouts for those of us who aren’t willing to turn their kitchen into a laboratory?

With the 3-day-old broccoli sprouts have the optimal yields study as a reference, minimum values of the six broccoli cultivated varieties studied were:

  1. Lowest weight of 100 seeds: .33 grams.
  2. 100 of that cultivar’s 3-day-old sprouts weighed 1.55 grams.
  3. A different cultivar had the lowest total phenolics (gallic acid equivalents): 0.94 mg per gram of 3-day-old sprouts.
  4. That second cultivar also had the lowest total flavonoids (rutin equivalents): 1.02 mg per gram of 3-day-old sprouts.
  5. A third cultivar had the lowest weight of sulforaphane in its seeds: 2.43 mg per gram of seeds.
  6. Reduction of sulforaphane content during germination from seeds to 3-day-old sprouts is evident from the below chart. The only 3-day-old sprout chart number the study provided was the best case, though. I requested the study data, but in the meantime..I enlarged the chart, measured the worst cultivar as a 8.5 cm bar where the seed bar was 16 cm. Its 3-day-old sprout sulforaphane yield was ~53% of its seed yield.

sprout ages 1B

Another caveat for contents: Researchers had to process broccoli seeds and 3-day-old broccoli sprouts in order to make measurements. Total phenolics and total flavonoids may not have been affected by processing. However, processing generated sulforaphane.

Broccoli seeds and 3-day-old broccoli sprouts contain little or no sulforaphane. They have glucoraphanin and myrosinase enzyme which are structurally separated. Disturbing their cells mixes the two, and the enzyme hydrolyzes glucoraphanin and other glucosinolates into sulforaphane and other healthy compounds.

I start out each daily batch with two tablespoons of broccoli seeds of an unknown cultivar. I counted 812 broccoli seeds in a teaspoon (yes, I did), and multiplied by 6 for 4,872 total seeds.

Runt-of-the-litter calculations for the six broccoli cultivars studied are:

  • (4,872 / 100) x .33 g = 16.08 10.7 g x 2 = 21.4 g broccoli seeds [my measurements];
  • (4,872 / 100) x 1.55 g = 75.52 65.5 x 2 = 131 g 3-day-old broccoli sprouts [my measurements];
  • 75.52 131 g x 0.94 mg per gram of 3-day-old sprouts = 70.99 123.1 mg total phenolics [worst-case];
  • 75.52 131 g x 1.02 mg per gram of 3-day-old sprouts = 77.03 133.6 mg total flavonoids [worst-case];
  • 16.08 21.4 g broccoli seeds x 2.43 mg per gram of seeds = 39.07 52.0 mg sulforaphane [worst-case].

I’ll use the Microwave broccoli to increase sulforaphane levels study as a reference for an alternate sulforaphane calculation. The cultivar wasn’t mentioned, only that it was ordinary broccoli purchased in Silver Spring, Maryland, grocery stores.

One assumption is that microwaving broccoli sprouts will have the same effects as microwaving broccoli florets to increase sulforaphane content. Are the similarities between broccoli floret and broccoli sprout characteristics enough to say whether or not that’s a valid assumption?

Another assumption is that there’s no beginning amount of sulforaphane in 3-day-old broccoli sprouts. Microwaving them on full power to 60°C produces all of the sulforaphane.

A third assumption is that the sulforaphane increase from .22 to 2.45 µmol / g of the tested broccoli florets fairly represents other broccoli florets.

((2.45 – .22 µmol / g)

x 75.52 131 g 3-day-old broccoli sprouts)

/ 5.64 μmol conversion of amount to weight

= 29.86 51.8 mg sulforaphane

Sulforaphane: Its “Coming of Age” as a Clinically Relevant Nutraceutical in the Prevention and Treatment of Chronic Disease concluded:

“The daily SFN [sulforaphane] dose found to achieve beneficial outcomes in most of the available clinical trials is around 20-40 mg.”

My daily consumption of broccoli sprout compounds is greater than this clinical trial range, with evidenced calculations of 29.86 51.8 ≈ 30 52 mg sulforaphane from microwaving 131 g of 3-day-old broccoli sprouts. I’ll include this estimate in Week 8 of Changing to a youthful phenotype with broccoli sprouts.

Microwave broccoli to increase flavonoid levels demonstrated 108.5% to 129.8% increases in quercetin and kaempferol levels from microwaving ordinary broccoli purchased in a Beltsville, Maryland, grocery store. I won’t use their method of a 1200W microwave on full power for one minute, but microwaving may be expected to increase the worst-case calculation of 77.03 133.6 ≈ 77 134 mg total flavonoids.

Broccoli seed suppliers are missing a marketing opportunity by not specifying their cultivars. They could be advertising specific benefits, etc.

The first study showed that sulforaphane weights vary widely among broccoli cultivars, from the 2.43 mg used above to 12.07 mg per gram of seeds. The best-case would be 21.4 g broccoli seeds x 12.07 mg per gram of seeds = 258.3 mg sulforaphane!

If your product was almost 5 times better than a competitor’s, why wouldn’t you advertise it?

Part 3 of Rejuvenation therapy and sulforaphane

gettingwell4 2-3 stars Required further work, Brain development, Cerebellum, Cerebrum, Epigenetics, Hippocampus, Hypothalamus, Limbic system, Lower brain, Memory, Stress, Telomeres , , , , , ,

Part 1 focused on the study’s clinical biomarkers. Part 2 highlighted its epigenetic clocks. Now we’ll look at rejuvenation of cognitive function.

Charts for this study’s most relevant human aging applications – measured by the new human-rat relative biological age clock – were in supplementary data due to combining study untreated tissue samples into clock training data. Reanalyses showed:

“Using the final version of the epigenetic clocks, we find that treatment effects become even more significant especially for the hypothalamus.”

Human-rat relative clock percentages of rejuvenation were:

  • “Blood 70.6%
  • Liver 79.4%
  • Heart 61.6%
  • Hypothalamus 20.9%”

The Discussion section addressed hypothalamus rejuvenation:

“Why does plasma fraction treatment not reduce brain epigenetic age by the same magnitude as it does other organs? We can only begin to address this question after having first understood what epigenetic aging entails.

As it stands, our knowledge in this area remains limited, but it is nevertheless clear that:

  1. Epigenetic aging is distinct from the process of cellular senescence and telomere attrition,
  2. Several types of tissue stem cells are epigenetically younger than non-stem cells of the same tissue,
  3. A considerable number of age-related methylation sites, including some clock CpGs, are proximal to genes whose proteins are involved in the process of development,
  4. Epigenetic clocks are associated with developmental timing, and
  5. Relate to an epigenomic maintenance system.

Collectively, these features indicate that epigenetic aging is intimately associated with the process of development and homeostatic maintenance of the body post-maturity.

  • While most organs of the body turnover during the lifetime of the host, albeit at different rates, the brain appears at best to do this at a very much slower rate.
  • While most tissues harbor stem cells that are necessary for replenishment and turnover, stem cells in adult brain have only been detected in a defined and very limited area of the subventricular zone, olfactory bulb (in rats), hippocampus and hypothalamic proliferative region.

As such, if plasma fraction treatment’s rejuvenating effect is:

  • Mediated through the process of development and
  • Involves tissue stem cells

then its effect on epigenetic age of the brain would appear to be modest, which indeed it does.

It is to be noted however, that improving brain function does not depend on neurogenesis as much as it does on synapse formation and factors such as NMDA receptors, which decline in density with age.

Assessment of plasma fraction treatment on cognitive function (learning and memory). Rats were subjected to Barnes maze test – nine consecutive days of test where the time (in seconds) required by rats to find the escape hole (latency) was recorded and plotted. Error bars depict 2 standard errors.

Within a month of plasma fraction treatment, rats exhibited significantly reduced latency to escape, i.e., they learned and remembered better. After the second month, treated rats began with a slightly reduced latency period compared to untreated old rats, and once again, they learned much faster than the latter.

By the third month, it was clear that treated rats remembered the maze much better than untreated ones even from the first day of test as their latency period was significantly reduced. By the end of the test period, their latency was similar to that of young rats. This feature was sustained and repeated in the fourth month.”

Not sure why there’s a 62-day gap between “Second month” and “Third month.” Maybe it had something to do with “First month” starting 10 days after the first treatment and “Third month” similarly starting 13 days after the second treatment?

Many of us know older people who lived well past their time of good cognitive function:

  • We see how they’re helpless and dependent; and
  • We see how others take advantage of them in their morbidity phase, where healthspan stops increasing but lifespan continues.

We can make personal plans for that day, sure. But let’s also put some urgency into applying this study’s new human-rat relative biological age clock, and make:

“A step change in aging research. Although conservation of aging mechanism could be equally deduced from the existence of multiple individual clocks for other mammals (mouse, dog), the single formula of the human-rat clock that is equally applicable to both species effectively demonstrates this fact.”

Week 7 of Changing to a youthful phenotype with broccoli sprouts

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

To follow up Week 6 of Changing an inflammatory phenotype with broccoli sprouts:

1. I changed this week’s title as a result of reading A rejuvenation therapy and sulforaphane study. It wasn’t about sulforaphane, but its clinical findings had commonalities with this broccoli sprouts effort. It’s become my blog’s most popular post, read by people in 50+ countries.

A close second is An environmental signaling paradigm of aging. The current study’s lead laboratory researcher presented his view five years ago on where aging evidence was pointing.

Part 2 of Rejuvenation therapy and sulforaphane better curated this study’s innovative epigenetic clock results. There are no sulforaphane clinical trials that also use epigenetic clocks.

What effects may broccoli sprout compounds have on human aging? With this new human-rat relative biological age clock, researchers can get reliable answers from rat studies, with human clinical trials needed only to confirm those findings!

2. This week I found out that exercising control over my charges to protect them from disease was counterproductive. I exposed them to harm, destroyed their community, and stunted their growth by forcing them to distance from each other for their own good.

Am I a politician, an unelected bureaucrat, or some other form of busybody? No. I admit my mistakes right away, I apologize, then I immediately try to do better.

A proper context:

  • In Week 2 I switched from sprouting trays with 1/16″ high ridges in the bottom to Russian-doll bowls. That solved a problem of excess moisture, with which broccoli sprouts don’t do well but bacteria do.
  • In Week 3 I rotated in the next larger sized bowl to replace the smallest bowl. My thought was that Day 3 broccoli sprouts were too crowded to dry in the smallest bowl.
  • At the end of Week 5 I doubled my starting amount of broccoli seeds from one to two tablespoons. To accommodate that increase, I again rotated in the next larger size bowl.

Starting in Week 6, I had uneven batch yields. The two larger bowls yielded noticeably fewer sprouts than did batches in the two smaller bowls.

What did bowl size have to do with yield?

Nothing, it was me. Turned out I’d neglected Plant Care 101 instructions to provide adequate moisture.

After rinsing, straining, and wicking out excess moisture with a paper towel twice daily, I then spread out seeds and sprouts to prevent problems with excess moisture. Broccoli seeds and sprouts in the two larger bowls were more separated than in the two smaller bowls.

All of which led to moisture levels that were inadequate for broccoli seeds and sprouts. All batches sprouted less well than their potential yield. The larger the bowl, the more my behavior adversely affected the batch.

Here’s what Day 2 and Day 3 yields were with my previous practices.  Batch volume of Day 2 in the smaller bowl was larger than Day 3!

I changed practices to group broccoli seeds and sprouts together at a step where I used to spread them out. Here’s the same bowl with my current practice, but at Day 2. It’s a larger volume than the previous practice’s Day 2:

I’ll guess that batch yield volumes have improved by 75%. I increased distilled water from 100 ml to 175 ml before microwaving since 100 ml no longer completely immersed increased Day 3 broccoli sprout volume. My 1000W full power microwave time concomitantly increased from 45 seconds to 65 seconds to achieve 58°C.

Better-developed batches also taste better. I still mix in mustard and eat Day 3 broccoli sprouts with other food.

3. My sulforaphane intake has probably decreased with current practices. 3-day-old broccoli sprouts have the optimal yields study said:

“Although germination reduces SF [sulforaphane] yield to some extent, it is beneficial to formation and accumulation of total phenol and flavonoids, ensuring health properties of sprouts. SF contents in sprouts were 46% – 97% of seeds, whereas TP [total phenolic] and TF [total flavonoid] contents in sprouts were 1.12 – 3.58 times higher than seeds among [broccoli] varieties.”

I’m not concerned about less sulforaphane with a two tablespoons starting amount of broccoli seeds. Even a one tablespoon starting amount yields 60 g of broccoli sprouts, twice that of Effects of long-term consumption of broccoli sprouts on inflammatory markers in overweight subjects. See our discussion in Understanding a clinical trial’s broccoli sprout amount.

4. Another week of no inflammatory problems after four-to-six-mile-long beach walks. I’m not pushing myself, just walking often, and working out my upper body every fourth day.

I emphasize eccentric motions in upper body workouts. I haven’t curated the below 2019 papers although they’re informative:

I don’t expect recovery times from workouts to shorten. What’s an appropriate exercise recovery time? found with 26.5 ± 6.5 year-old male subjects that even three days wasn’t enough time for biceps brachii to fully recover from eccentric exercise.

5. During Friday’s walk I accidentally startled a large turkey hen who was on the ground, and she flew up on a fence. Can you see her moments before she hopped down?

Don’t have any idea what bugs a turkey found attractive near a beach.

See Week 8 of Changing to a youthful phenotype with broccoli sprouts for follow ups.