A broccoli sprouts study that lacked evidence for human applicability

December 12, 2020December 13, 2020 gettingwell4 0-1 star Wasted resources, Brain development, Epigenetics, Telomeres Control group, Critical period, DNA methylation, Embryo development, Genetic factors, Infants

A 2020 study Combined Broccoli Sprouts and Green Tea Polyphenols Contribute to the Prevention of Estrogen Receptor–Negative Mammary Cancer via Cell Cycle Arrest and Inducing Apoptosis in HER2/neu Mice (not freely available) conclusion was:

“Lifelong BSp [broccoli sprouts] and GTP [green tea polyphenol] administration can prevent estrogen receptor–negative mammary tumorigenesis through cell cycle arrest and inducing apoptosis in HER2/neu mice.”

These researchers had unaddressed insufficiencies in this study that were also in their 2018 study as curated below. The largest item that required translation into human applicability was rodent diet content of 26% “broccoli sprout seeds.”

You may be surprised to read the below previous study’s unevidenced advice to eat double the weight of broccoli sprouts that I eat every day. You won’t be surprised that it’s not going to happen. Especially when no alternatives were presented because rodent diet details weren’t analyzed and published.

Sulforaphane is an evolved defense mechanism to ward off predators, and eating it is evolutionarily unpleasant. Will people in general and pregnant women in particular eat a diet equivalent to 26% “broccoli sprout seeds?”

Where were peer reviewer comments and researcher responses? Are these not public as they are by all Open Access journals hosted on https://www.mdpi.com/?

Sponsors and researchers become locked into paradigms that permit human-inapplicable animal research year after year. What keeps them from developing sufficient human-applicable evidence to support their hypotheses?

This 2018 Alabama rodent study investigated the epigenetic effects on developing breast cancer of timing a sulforaphane-based broccoli sprouts diet. Timing of the diet was as follows:

Conception through weaning (postnatal day 28), named the Prenatal/maternal BSp (broccoli sprouts) treatment (what the mothers ate starting when they were adults at 12 weeks until their pups were weaned; the pups were never on a broccoli sprouts diet);
Postnatal day 28 through the termination of the experiment, named the Postnatal early-life BSp treatment (what the offspring ate starting at 4 weeks; the mothers were never on a broccoli sprouts diet); and
Postnatal day 56 through the termination of the experiment, named the Postnatal adult BSp treatment (what the offspring ate starting when they were adults at 8 weeks; the mothers were never on a broccoli sprouts diet).

“The experiment was terminated when the mean tumor diameter in the control mice exceeded 1.0 cm.

Our study indicates a prenatal/maternal BSp dietary treatment exhibited maximal preventive effects in inhibiting breast cancer development compared to postnatal early-life and adult BSp treatments in two transgenic mouse models that can develop breast cancer.

Postnatal early-life BSp treatment starting prior to puberty onset showed protective effects in prevention of breast cancer but was not as effective as the prenatal/maternal BSp treatment. However, adulthood-administered BSp diet did not reduce mammary tumorigenesis.

The prenatal/maternal BSp diet may:

Primarily influence histone modification processes rather than DNA methylation processes that may contribute to its early breast cancer prevention effects;

Exert its transplacental breast cancer chemoprevention effects through enhanced histone acetylation activator markers due to reduced HDAC1 expression and enzymatic activity.

This may be also due to the importance of a dietary intervention window that occurs during a critical oncogenic transition period, which is in early life for these two tested transgenic mouse models. Determination of a critical oncogenic transition period could be complicated in humans, which may partially explain the controversial findings of the adult BSp treatment on breast cancer development in the tested mouse models as compared the previous studies. Thus long-term consumption of BSp diet is recommended to prevent cancers in humans.”

“The dietary concentration for BSp used in the mouse studies was 26% BSp in formulated diet, which is equivalent to 266 g (~4 cups) BSp/per day for human consumption. The concentration of BSp in this diet is physiological available and represents a practical consumption level in the human diet.

Prior to the experiment, we tested the potential influences of this prenatal/maternal BSp regimen on maternal and offspring health as well as mammary gland development in the offspring. Our results showed there was no negative effect of this dietary regimen on the above mentioned factors (data not shown) suggesting this diet is safe to use during pregnancy.”

I didn’t see where the above-labelled “Broccoli Sprout Seeds” diet content was defined. It’s one thing to state:

“SFN as the most abundant and bioactive compound in the BSp diet has been identified as a potent HDAC inhibitor that preferably influences histone acetylation processes.”

and describe how sulforaphane may do this and may do that, and include it in the study’s title. It’s another thing to quantify an animal study into findings that can help humans.

The study’s food manufacturer offers dietary products to the public without quantifying all contents. Good for them if they can stay in business by serving customers who can’t be bothered with scientific evidence.

Any difference between the above-labelled “Broccoli Sprout Seeds” and broccoli seeds? Where was any evidence that “Broccoli Sprout Seeds” and SPROUTED “Broccoli Sprout Seeds” were equivalent per this claim:

“Equivalent to 266 g (~4 cups) BSp/per day for human consumption. The concentration of BSp in this diet is physiological available and represents a practical consumption level in the human diet.”

To help humans, this animal study had to have more details than the food manufacturer provided. These researchers should have either tasked the manufacturer to specify “Broccoli Sprout Seeds” content, or contracted out analysis if they weren’t going to do it themselves.

Regarding timing of a broccoli sprouts diet for humans, this study didn’t provide evidence for recommending:

“Long-term consumption of BSp diet is recommended to prevent cancers in humans.”

http://cancerpreventionresearch.aacrjournals.org/content/early/2018/05/15/1940-6207.CAPR-17-0423.full-text.pdf “Temporal efficacy of a sulforaphane-based broccoli sprout diet in prevention of breast cancer through modulation of epigenetic mechanisms”

Sprouting oats

December 12, 2020December 26, 2020 gettingwell4 4-5 stars Advanced knowledge, Epigenetics, Immune system, Stress Control group, Embryo development, Genetic factors, Neurodegenerative disease

Three 2020 studies investigated properties of sprouted oats. This first study compared one oat cultivar’s seed and sprout contents for phenolic compounds, and evaluated oat sprouts’ protection against developing colon cancer:

“The purpose of this investigation was to evaluate whether sprouted oats (SO) of the Turquesa variety still possessed effective physiologically bioactive compounds, i.e., phenolics, flavonoids, AVAs [avenanthramides], and phytosterols, and whether it exerted antioxidant and anti-inflammatory effects, as well as the capacity to improve relevant intestinal parameters, in an AOM [azoxymethane] / DSS [dextran sulfate sodium]-induced CRC [colorectal cancer] mouse model.

Suboptimal intake of whole grains (38 g/d) was associated with CRC burden across 16 European countries. An optimal intake of 50–100 g/d was considered in our study to establish the dose administered in the AOM/DSS-induced CRC mouse model (75 g/d).

Seeds (100 g) were soaked in distilled water for 12 h then watered daily. Temperature and relative humidity were set at 25 °C and 60%. Germination was performed in darkness for five days. Germination percentage was determined based on total number of fully emerged seedlings.

We reached 100% of germination and a radicle length of 6.47 ± 0.22 cm. Sprouts were dried at 50 °C for 12 h, milled to a particle size of 0.5 mm, and stored at 4 °C until analyses.

Protein and lipid contents were higher in SO, whereas carbohydrate and ash contents were lower. A more than four-fold increase [0.64 mg/g to 2.79 mg/g] in TPC [total phenolic compounds] was obtained after five days.

We identified AVA-D as the most abundant AVA, followed by AVA-L, which had not been reported as one of the three most abundant AVAs in other oat varieties. Of the three most abundant AVAs previously reported, only AVA-B had a higher abundance in germination.

Phytic acid, an antinutritional compound present in oats, was 10 times lower in oat sprouts. Phytic acid has its content decreased by 15%–35% during even a short three-day germination due to activation of phytase activity. Although high doses of phytic acid inhibit absorption of metals and minerals in humans, it has been observed that, in small doses, it can function as a protective factor in several chronic degenerative diseases.

Mice in groups 3 and 4 were gavaged every morning with phenolic-AVA extract (0.084 mg GAE) and 30 mg of SO, respectively. We observed a mild anti-inflammatory effect of SO and AVA treatments, and a reduced adenocarcinoma incidence of 52.5% and 21.3%, respectively.

SO was more efficient in activating the Keap1-Nrf2 signaling pathway compared to treatment with AVA. Oat phenolic compounds together with β-glucans may be acting synergistically, thus offering greater protection for cancer prevention and treatment.”

https://www.mdpi.com/2304-8158/9/2/169/htm “Chemopreventive Effect of the Germinated Oat and Its Phenolic-AVA Extract in Azoxymethane/Dextran Sulfate Sodium (AOM/DSS) Model of Colon Carcinogenesis in Mice”

The supplementary material developed this oat cultivar’s seed and sprout profiles for 138 phenolic compounds. It measured C-type AVAs, but not A-type AVAs.

This was my model study for Sprouting whole oats.

A second study was reviewed in Eat oats today! and repeated here:

“The first evaluation of anti-inflammation effects of A-type AVAs was published from our own group. Fifteen A-type AVAs from commercial sprouted oat products interacted with lipopolysaccharide-induced nitric oxide production and iNOS expression.”

https://pubs.acs.org/doi/full/10.1021/acs.jafc.9b06812 “Quantitative Analysis and Anti-inflammatory Activity Evaluation of the A-Type Avenanthramides in Commercial Sprouted Oat Products” (not freely available)

Oat variety and sprout age weren’t available for the six sprouted oat products tested, so oat seed-to-sprout comparisons weren’t possible. A-type AVA comparisons among products were performed, but weren’t meaningful due to unknown varieties, ages, product processing, and storage.

A third study compared four grains’ sprouted and unsprouted contents:

“Seeds were soaked at 25°C in 1 L of distilled water for 20 (brown rice), 12 (sorghum and millet) and 8 h (oat), respectively. Hydrated grains were allowed to germinate with layering over wet cellulose pads in a humid chamber for 60 h at 25°C (oat seeds) or 30°C (brown rice, sorghum, and millet seeds) with 95% relative humidity.

All seeds derived from brown rice and oat were germinated after 48 h in the humid chamber. Germinated grains were dried at 50°C until reaching a moisture content of 10%. Sample seeds were milled to fine flour, screened through a 100-mesh sieve and stored at 4°C for further analysis.

After 60 h of germination, sprout length in sorghum and millet ranged from 8 to 24 mm, while sprouts obtained from brown rice and oat ranged from 3 to 6 mm.

Compared to raw flours, germinated flours derived from brown rice, sorghum, and millet had lower gelatinization enthalpy, whereas germinated oat flour showed higher gelatinization enthalpy.

During germination, enzymes are activated, catalyzing starch degradation, which may disrupt the double helical structure of starch. Consequently, less energy is required to unravel and melt double helices of starch in germinated flours. The increase in gelatinization enthalpy of germinated oat flour may be due to dissolution of hydrolyzed starch granules during germination.”

https://link.springer.com/article/10.1007%2Fs10068-020-00770-2 “Influence of germination on physicochemical properties of flours from brown rice, oat, sorghum, and millet” (not freely available)

The first study sprouted oats for five days to full germination and a minimum radicle length of 6.25 cm. The third study sprouted oats to full germination in 60 hours and a 3 mm minimum total length.

At the same 25°C, with 60% relative humidity and daily watering, it took 120 hours to achieve full germination. With 95% relative humidity, it took half that time.

Was humidity a relevant difference in oat sprout growth? Would Choyang variety oat sprouts increase their minimum 3 mm total length more than 20 times between Hours 60 and 120 to match the minimum Turquesa radicle length?

This is a count of PubMed “oat sprout” search results, 20 results total:

A “broccoli sprout” search returned 648 results. Is oat sprout research just getting started?

Part 2 of The transgenerational impact of Roundup exposure

December 11, 2020December 13, 2020 gettingwell4 4-5 stars Advanced knowledge, Epigenetics, Memory, Stress Control group, DNA methylation, Embryo development, Inherited disease

This 2020 study followed up The transgenerational impact of Roundup exposure using the Washington State Unversity research group’s most recent methodology in DEET and permethrin cause transgenerational diseases:

“The herbicide glyphosate has been shown to promote epigenetic transgenerational inheritance of pathology and disease in subsequent great-grand offspring (F₃ generation). The current study was designed to identify epigenetic biomarkers for glyphosate-induced transgenerational diseases using an epigenome-wide association study.

Pathologies investigated included prostate disease [13 of 44 subjects], kidney disease [11 of 44], obesity [19 of 45], and presence of multiple disease [10 of 45]. Sperm were collected from F₃ glyphosate lineage males and used to identify specific differential DNA methylation regions (DMRs) and differential histone retention sites (DHRs).

The number of DHRs were less than the number of DMRs, and DHRs were found to have disease specificity. The combination of DMRs and DHRs is anticipated to facilitate pathology diagnosis.

Low sample number is a limitation in the current analysis. Potential higher variability in data needs to be considered.

This is one of the first observations of DHRs as potential biomarkers for disease. The current study used glyphosate induction of transgenerational disease as a proof of concept such environmental biomarkers can be identified and potentially used as diagnostics for disease susceptibility in the future.”

https://www.tandfonline.com/doi/full/10.1080/15592294.2020.1853319 “Epigenome-wide association study for glyphosate induced transgenerational sperm DNA methylation and histone retention epigenetic biomarkers for disease”

Reverberations, harmonics, history

December 10, 2020December 10, 2020 gettingwell4 2-3 stars Required further work, Beliefs, Memory Reputations

Catching up with Martin Armstrong from 2012:

“Corruption within the Roman Republic was certainly at its peak during the first century BC. There was a brewing debt crisis in Rome and the oligarchy was determined to keep power at any cost. Corruption was so widespread that interest rates doubled from 4% to 8% for the elections of 54 BC because there was so much bribery going on to gain votes.

Caesar was clearly a Popularis, a man of the people who stood against the corruption of the Republic. Like today, we have no real voting control over the fate of the nation. Those who are in charge of the political machine control the real political state.

Caesar knew who his enemies truly were. He clung to his belief that if the majority of the Senate were free of the Oligarchy of Cato and Cicero, they would surely see the light. To persuade them, Caesar wrote his seven books on his truly remarkable conquest of Gaul.

Cato and his Oligarchy were so intensely anti-Caesar that they were willing to do anything to anybody. But this was a moment in time where the corruption had simply gone too far.

By September 29th, 51 BC, Caesar ran out of civilized options. Crossing the Rubicon became the only option.

Janus was the symbol of a cyclical change, the departing of one era and the birth of another. His shrine consisted of two doorways that traditionally were left open in time of war and kept closed when Rome was at peace. Leaving the doors open in time of war symbolized the new era that was possible.

Property values were collapsing. Debts were excessive. Those who held mortgages refused to accept just the property back.

Caesar dealt with this major extraordinary situation in a truly astonishing manner, realizing that assets and money are in a union of opposing forces, yet bound together. Value of property is not a constant relationship for money itself is not like a ruler.

Money is more akin to a rubber band even when it may be gold or silver. Money is like everything else – subject to the whims of supply and demand.

The economy is a dynamic relationship between everything with no real constant. We are at a tremendous disadvantage because we have grown up thinking in a flat linear world that does not exist. We limit ourselves by thinking in money,

Caesar explained that he had to borrow to fund the war and it was unethical for him to cancel all debts since he himself would benefit. Generals come and go, but true economic reformers of the state to save the nation are rare indeed. Caesar paid for his economic reform with his life.

There can be no greater example of political corruption that required desperate reform than the calendar. Caesar replaced the typical lunar year and introduced his new calendar based on 365¼ solar days on January 1st, 45 BC.

Sulla [138 – 78 BC] was a highly original, gifted and skillful general, never losing a battle. His rival described Sulla as having the cunning of a fox and the courage of a lion – but that it was the former attribute that was by far the most dangerous. This mixture was later referred to by Machiavelli in his description of the ideal characteristics of a ruler.

Sulla was more interested in retaining institutes of government while eliminating people occupying them whereas Caesar was far more compelled to act to restore institutions and to spare people, even his more threatening enemies. These are not actions of a man interested in personal power, but a man interested in saving his country.

You live in an oligarchy no different today than what Caesar faced back then. One maxim always holds true; Absolute power, corrupts absolutely!”

It appears that only Julius Caeser ever understood

100-44 BC

A normal distribution

Does sulforaphane reach the colon?

December 8, 2020April 21, 2021 gettingwell4 2-3 stars Required further work, Epigenetics, Immune system

This 2020 study subject was antimicrobial activity of sulforaphane:

“This study explored the role that digestion and cooking practices play in bioactivity and bioavailability, especially the rarely considered dose delivered to the colon.

A broccoli powder soup was prepared which contained 26.5 µmol of sulforaphane per 200 ml portion. Addition of 2% mustard seed powder at the cooling stage of the soup preparation process (~ 60 °C) increased the level of sulforaphane by nearly fourfold, 102 µmol per 200 ml.

Recovery of sulforaphane in ileal fluids post soup consumption was < 1% but the addition of mustard seeds increased colon-available sulforaphane sixfold. Analysis of glucosinolates composition in ileal fluids revealed noticeable inter-individual differences.

Consumption of sulforaphane-enriched broccoli soup may inhibit bacterial growth in the stomach and upper small intestine, but not in the terminal ileum or the colon.”

https://link.springer.com/article/10.1007/s00394-020-02322-0 “Sulforaphane-enriched extracts from glucoraphanin-rich broccoli exert antimicrobial activity against gut pathogens in vitro and innovative cooking methods increase in vivo intestinal delivery of sulforaphane”

My son has often asked me about adding mustard seeds to broccoli sprouts. Papers citing one of this study’s coauthors’ series of mustard seed studies include:

I bought a 74 gram container of mustard seeds at the local grocery store, and ground down a third as pictured. A level scoop of mustard seed powder weighs 1.5 grams.

1.5 g divided by my twice-daily 65 g of microwaved broccoli sprouts = 2%, matching this study’s methods. That’s a 24-day mustard seed supply for $2.19.

I’ll add mustard seed powder immediately after microwaving broccoli sprouts when they’re ≤ 60°C (140°F). Allowing the mixture to process for five minutes potentially facilitates myrosinase hydrolization of glucoraphanin and other glucosinolates into healthy isothiocyanate compounds.

Other aspects of this study:

1. I don’t consider overcooking broccoli an “innovative cooking method.” It’s more like researchers creating an effect in order to publish “increased the level of sulforaphane by nearly fourfold” which was presented numerically and emphasized twice in text.

2. A perspective on these types of studies from Epigenetic mechanisms of muscle memory:

“Underpowered studies may only be useful to check if the experiment works, but not as much for testing and estimating effects.”

3. I didn’t agree with this study’s treatment of individual differences.

I read three other papers’ study design recommendations for researchers regarding inter-individual variability, but didn’t see markedly better ideas. Most of their verbiage concerned how to reduce heterogeneous effects rather than to understand individual causes and signals.

Where are thoughtful counters to meaningless averages / standard deviations / p values?

4. “Addition of mustard seeds increased colon-available sulforaphane sixfold” was presented numerically and emphasized thrice in text. Too often for a n=11 study.

What needed further explanations were detailed causes for each individual’s responses or lack thereof. Stratifying subgroups into unresponsive:

What happened in Subjects 6’s and 10’s lives to make them unresponsive to any sulforaphane dose?
Were Subjects 1, 2, 5, and 7 instances of zero sulforaphane actually errors in measuring or processing? If not, what were individual causes for instances of no response?

And responsive:

Were Subjects 4, 8, 9, 11, and 12 averages meaningful? Excluding Subject 4’s 3.14 μmol, was the four remaining subjects’ 0.19 to 0.63 μmol average 332% increased response meaningful when the sulforaphane dose increased 392%?
What caused Subject 4’s 872% increased response when the sulforaphane dose increased 392%?

Findings of sulforaphane in 11 g broccoli powder not reaching the colon may not apply to twice-daily 65.5 g microwaved broccoli sprouts due to mass and quantity differences. Broccoli sprouts definitely pass into the colon, like any other fibrous vegetable. Unhydrolyzed products are hydrolyzed by microflora there.

I create sulforaphane from broccoli sprouts shortly before eating them, and don’t depend on metabolism after the stomach to produce isothiocyanates. Did this study’s findings assist with that effort?

Eat oats today!

December 6, 2020December 6, 2020 gettingwell4 4-5 stars Advanced knowledge, Acetylcholine, Dopamine, Epigenetics, Hippocampus, Immune system, Limbic system, Memory, Neurochemicals, Serotonin, Stress Brain neurons, Control group, Genetic factors, Neural plasticity, Neurodegenerative disease

This 2020 food chemistry review provided phenolic-compound reasons to eat oats:

“Phenolamides result from the conjugation of hydroxycinnamic acids with amines. These products contain a variety of metabolic, chemical, and functional capabilities due to the large number of possible combinations among the parent compounds.

Of the currently known phenolamides, the most common are avenanthramides (AVAs), which are unique in oats. AVAs possess anti-inflammatory, anti-itch, anti-atherosclerosis, antioxidant, anti-cancer, anti-obesity, anti-fungal, anti-microbial, and neuroprotective properties.

Twenty-nine C-type AVAs have been identified in oats, and twenty-six A-type AVAs.

C-type AVAs in commercially available oat products range from 36.49-61.77 mg/kg (fresh weight).

A-type AVAs represent approximately 22.5% of total AVA levels in regular oats and 24.7-33.0% in commercial sprouted oats.

Steeping raw groats increased AVA concentrations.”

These reviews were referenced:

“Since publication of these two reviews, a few new studies reported AVAs’ beneficial health effects, mainly related to their anti-inflammatory and anti-cancer activities. AVAs can:

Significantly decrease IL-6, IL-8, and MCP-1 in endothelial cells;

Inhibit IL-1β- and TNF-α-induced NF-κB activation; as well as

Expression of adhesion molecules; and

Adhesion of monocytes to endothelial cell monolayer.

In 2020, the first evaluation of anti-inflammation effects of A-type AVAs was published from our own group. Fifteen A-type AVAs from commercial sprouted oat products interacted with lipopolysaccharide-induced nitric oxide production and iNOS expression.

Colloidal oatmeal’s natural components, AVAs, help to restore and maintain skin barrier function. AVAs are safe, well tolerated, and can be effective as adjuvant treatment in atopic dermatitis.

In one mouse model, a C-type AVA was able to mitigate many adverse effects of Alzheimer’s Disease. It restored hippocampal long-term potentiation and synaptic function, enhanced memory function, suppressed pro-inflammatory cytokines TNF-α and IL-6 levels, reduced caspase-3 levels, and increased pS9GSK-3β and IL-10 levels.

AVAs downregulated expression of hTERT and MDR1, pro-survival genes for cancer cells, and COX-2 mRNA and PGE2 levels, known pro-inflammatory markers. AVAs induced apoptosis by activating caspases 8, 3, and 2.”

https://pubs.acs.org/doi/10.1021/acs.jafc.0c02605 “The Chemistry and Health Benefits of Dietary Phenolamides” (not freely available)

Hadn’t thought about sprouting oats before this paper.

Measuring one dimension of health

December 5, 2020February 24, 2021 gettingwell4 0-1 star Wasted resources, Epigenetics, Immune system

This 2020 human study asserted:

“Our data provides the first epidemiological evidence supporting evidence obtained in preclinical models of metabolic syndrome and NAFLD that demonstrated hepatoprotective effects of phenolic acids.

High dietary intake of total phenolic acids is associated with a lower prevalence of non-alcoholic fatty liver disease and insulin resistance.

High intake of hydroxybenzoic acids, a class of phenolic acids, is associated with a lower prevalence of steatosis and clinically significant fibrosis.

High intake of hydroxycinnamic acids, another class of phenolic acids, is associated with a lower prevalence of insulin resistance.

Data on polyphenol content in foods was obtained from the Phenol-Explorer database (www.phenol-explorer.eu).”

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7078532/ “Higher phenolic acid intake independently associates with lower prevalence of insulin resistance and non-alcoholic fatty liver disease”

It’s a bad weather day, so I investigated the phenolics database and ran some numbers:

Phenolic contents of all the other food I eat is 9% of my coffee-and-tea 1,975 mg total phenolics. Microwaved broccoli sprouts contribute half of that 9%.

Subjects were grouped according to whether their phenolics daily intake was over 221 mg or not. The over 221 mg group drank 5 cups of coffee a day, whereas the other group drank 1 cup.

According to a phenolics dimension of health, all these researchers needed to do was ask subjects about their daily coffee intake. But then the study would be over, with few “is associated with” findings.

Do humans avoid insulin resistance and non-alcoholic fatty liver disease by drinking more than one cup of coffee and tea? Is an answer available from real people, not just from a statistics package?

A 2020 study that primarily sourced a database last updated in June 2015 selected a fertile ground for later hypothesis-seeking.

Ignoring subsequent research helped when staking a claim of first for whatever niche provided a publication opportunity.

I didn’t upload a screenshot of the Excel workbook with entries for pictured items I eat every day. That June 2015 database was incomplete with respect to current science here in December 2020.

See Eat oats today! for current examples of phenolic compounds in my daily 81 grams of steel-cut oats.

In memory of freedom through February 2020

December 3, 2020December 6, 2020 gettingwell4 Just me

Photos taken in Milan and Lake Como, Italy, February 22-23, 2020. The same weekend during which ten towns were closed south of the city.

As I said in Train your immune system every day! my traveling companion and I were likely exposed, yet had no symptoms after two weeks. Take responsibility for your one precious life.

Are humans more enlightened now regarding:

“Inalienable rights; that among which these are life, liberty, & the pursuit of happiness; that to secure these rights, governments are instituted among men, deriving their just powers from the consent of the governed.”

than in 1776?

Image from the US Library of Congress.

Have you given your silent obedient consent to the worldwide power grab?

Believe when I say

December 2, 2020January 26, 2021 gettingwell4 Just me

Clearing out the 2020 queue of interesting papers

December 1, 2020December 1, 2020 gettingwell4 2-3 stars Required further work, Acetylcholine, Brain development, Cerebrum, Cortisol, Dopamine, Epigenetics, Glutamate, Hippocampus, Hypothalamus, Immune system, Limbic system, Lower brain, Memory, Neurochemicals, Serotonin, Stress Antidepressants, Brain neurons, Control group, Critical period, DNA methylation, Embryo development, Genetic factors, Infants, Inherited disease, Neural plasticity, Neurodegenerative disease, Prefrontal cortex, Psychotropic medication

I’ve partially read these 39 studies and reviews, but haven’t taken time to curate them.

Early Life

Epigenetic clocks

Epigenetics

Aging

Broccoli sprouts

Does reprogramming signaling pathways create memories?

November 29, 2020February 10, 2023 gettingwell4 0-1 star Wasted resources, Epigenetics, Immune system, Memory, Stress

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

Zinc and broccoli sprouts – a winning combination

November 29, 2020August 7, 2021 gettingwell4 5+ stars Premium, Epigenetics, Immune system, Memory, Stress Control group

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

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

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

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

Heart mass was significantly lower in the IH-BSE/Zn group than in IH and IH-BSE groups. Heart mass / tibia length ratio was significantly lower in the IH-BSE/Zn group than in IH and monotherapy groups.

Treatment with BSE and/or Zn can ameliorate myocardial fibrosis associated with IH, to a certain extent, and combination therapy had the best antifibrotic effect among treatments.

BSE or Zn can significantly ameliorate myocardial inflammation induced by IH, but the combination provided a better anti-inflammatory effect.

We used 3-NT as an indicator of the severity of oxidative stress. 3-NT protein levels were significantly reduced in IH mice for all treatment groups, and reduction was greater in the combination treatment group.

Combination was more effective than monotherapies to activate Nrf2-mediated antioxidant function.

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

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

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

Week 34 of Changing to a youthful phenotype with broccoli sprouts

November 26, 2020August 27, 2023 gettingwell4 4-5 stars Advanced knowledge, Epigenetics, Immune system, Stress Control group, Reputations, Unconscious feelings, Unconscious memories

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

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

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

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

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

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

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

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

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

“I3C in broccoli was increased by 3.1, 9.1 and 1.9 folds respectively using blenders 1, 2 and 5 with microwaving.”
Microwave broccoli to increase flavonoid levels study design was “Broccoli florets (150 g) were put in a microwave safe bowl with a 1 tablespoon [15 ml] of water” and a 1200W microwave on full power for one minute. Although this may have produced temperatures > 60°C, flavonoid fresh weight contents increased > 30%:

“Microwaving may increase extractability and/or release from binding to other compounds as a result of matrix softening.“
Microwave broccoli to increase sulforaphane levels demonstrated significant differences for 475W (LL) and 950W (HL) power settings in glucoraphanin and sulforaphane dry weight amounts when broccoli florets were microwaved to the same temperatures. Compare white bar sulforaphane amounts for LL60 and HL60 (both 60°C), annotated as E and F:

“Microwave treatment causes a sudden collapse of cell structure due to the increase in osmotic pressure difference over vacuole membrane. Microwave irradiation might help to release more conjugated forms of glucosinolates and then get hydrolyzed by released myrosinase.“
Enhancing sulforaphane content confirmed the above 60°C finding with broccoli florets:

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

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

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

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

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

Part 2 of Eat broccoli sprouts for your eyes

November 22, 2020January 19, 2025 gettingwell4 4-5 stars Advanced knowledge, Epigenetics, Memory, Stress Brain neurons, Control group, Neurodegenerative disease

I was a little bothered by an unreferenced statement in Eat broccoli sprouts for your eyes that:

“Once AGEs are formed, most are irreversible.”

I searched curated 2020 studies for “revers” and found that recent blog studies favored reversibility of epigenetic changes 12-to-2. Do they reflect my selection bias, or is there something different about AGEs?

Let’s start with this statement:

“Although AGEs are irreversible adducts and cross-links in our tissues, these can be removed through different proteolytic capacities:

The ubiquitin proteasome system (UPS) – Ubiquitin is a protein that when conjugated to a protein substrate can facilitate degradation of that substrate by the proteasome. Obsolete or damaged proteins are tagged with ubiquitin and these ubiquitinated substrates are degraded by the proteasome. Operates mainly on soluble substrates.

Autophagy – Can operate on insoluble substrates, including organelles such as mitochondria. Autophagy requires macromolecular assemblies and organelles to identify, sequester, and eventually degrade substrates via the lysosome.

Unfortunately, the function of both proteolytic pathways declines with extensive glycative stress and upon aging in many tissues, resulting in intracellular accumulation of protein aggregates (also glycated conjugates) and dysfunctional organelles. This thwarts strategies to lower AGEs accumulation by boosting proteolytic capacities.”

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

So humans can remove irreversible AGE epigenetic changes as long as the individual isn’t too stressed or old? Studies from 2008 to 2012 were cited for the above statement and graphic.

Citation 211 Sulforaphane delays diabetes-induced retinal photoreceptor cell degeneration (not freely available) 2020 findings were instructive:

“SF [sulforaphane] can delay photoreceptor degeneration in diabetes. The underlying mechanism is related to:

Inhibition of ER [endoplasmic reticulum] stress;

Inflammation; and

Txnip [thioredoxin-interacting protein] expression through activation of the AMPK [adenosine 5′-monophosphate (AMP)-activated protein kinase] pathway.

Function of the retina in diabetic mice [DM] as determined by ERG [electroretinography].”

This chart demonstrated that preventing diabetes’ negative effects on retinal function (i.e. controls) was measurably better than trying to fix subjects’ vision after onset of diabetes. Are future choices of humans who give themselves this non-communicable disease also limited to addressing symptoms?

The AMPK pathway was previously mentioned in:

Reversal of aging and immunosenescent trends with sulforaphane:

“Dihydroxyvitamin D3 and sulforaphane are compounds that safely induce AMPK activation, and may have wide-ranging implications for both normal and pathological aging.”
Part 2 of Reversal of aging and immunosenescent trends with sulforaphane:

“NQO1 plays a key role in AMPK-induced cancer cell death 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. 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.”

This first example was vitamin D3’s separate yet connected signaling pathway that acts both additively and synergistically with broccoli sprout compound effects. The second was signaling pathways becoming cascadingly activated from sulforaphane’s main effect, Nrf2 signaling pathway activation.

Eat broccoli sprouts for your eyes

November 21, 2020February 26, 2021 gettingwell4 4-5 stars Advanced knowledge, Dopamine, Epigenetics, Immune system, Neurochemicals, Stress Brain neurons, Control group, DNA methylation, Genetic factors, Neurodegenerative disease

This 2020 review subject concerned a leading cause of blindness:

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

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

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

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

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

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

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

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

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

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

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

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

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

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

Continued in Part 2.