Back pain and advanced glycation end products (AGEs)

Two 2020 rodent studies investigated intervertebral disk degeneration, with the first on AGEs’ role:

“This study evaluated the role of AGEs and RAGE in driving early intervertebral disk (IVD) degeneration processes in mice. Aging and diabetes are associated with increased low-back pain and IVD degeneration, yet causal mechanisms remain uncertain. AGEs:

  • Accumulate in IVDs from aging;
  • Are implicated in diabetes-related disorders;
  • Alter collagen; and
  • Induce proinflammatory conditions.

A mixed population of 23 male and female wild type AC57BL/6J mice were each assigned to two isocaloric diet groups after weaning. They received either low-AGE chow containing 7.6 μg/mg AGE, or high-AGE chow containing 40.9 μg/mg AGE generated via high-temperature heating (NIH-31 open formula chow autoclaved for 30 minutes at 120°C [248° F]). This in vivo dietary model was previously shown to increase IVD AGE accumulation without systemic obesity or diabetes.

disc AGE damage

AGE accumulation leads to RAGE-dependent collagen disruption in the annulus fibrosus, and can initiate molecular and tissue level collagen disruption. Second harmonic generation (SHG) and collagen-hybridizing peptide (CHP) analyzes were sensitive to collagenous alterations at multiple hierarchical levels due to AGE.

These methods may be useful in identifying additional contributors to collagen damage in IVD degeneration processes.”

https://onlinelibrary.wiley.com/doi/10.1002/jsp2.1126 “Advanced glycation end products cause RAGE-dependent annulus fibrosus collagen disruption and loss identified using in situ second harmonic generation imaging in mice intervertebral disk in vivo and in organ culture models”

Other human studies found degenerative spine disorders start at detectable levels during adolescence. Those study designs didn’t trace disc degeneration to diet, though.


A second study was summarized in a conference I’m sure researchers would like to reconvene:

“Kritschil et al investigated the role of insulin-like growth factor 1 (IGF-1) signaling on progression of disc degeneration in aging mice. They showed that diminished IGF-1 bioavailability confers both beneficial effects of decreased disc cell senescence and extracellular matrix catabolism, whilst at the same time negatively impacting proteoglycan production.”

jsp21134-fig-0001-m

https://onlinelibrary.wiley.com/doi/10.1002/jsp2.1134 “Advancing basic and preclinical spine research: Highlights from the ORS PSRS 5th International Spine Research Symposium”

https://onlinelibrary.wiley.com/doi/10.1002/jsp2.1112 “Effects of suppressing bioavailability of insulin-like growth factor on age-associated intervertebral disc degeneration”

This study asserted:

“Despite some inconsistent findings on the role of IGF-1 among human centenarian and animal model studies, there is overwhelming evidence to support that disruptions to the IGF-1 signaling pathway promotes healthy longevity.”

See Take responsibility for your one precious life – DHEA for other evidence on IGF-1.


Spent a large part of this weekend reading abstracts and studies concerning diet interactions with spinal disc degeneration. This AGE study provided more evidence than others on these relationships.

I’ve eaten AGE-less chicken vegetable soup almost every day for two years:

  • 237 g chicken breast cubes, 179 g celery, and 262 g carrots in 1 cup Savignon Blanc get up to 100° C around 9 minutes initially, then again about 6 minutes after I add 1 quart chicken broth, then I turn off the Instant Pot.
  • I stir in 340 g mushrooms, 31 g garlic, and 387 g Roma tomatoes five minutes later at about 85° C, and they cool the soup down to around 70° C. I let it stew for another 15 minutes before eating half (1.5 quarts).
  • A 1.5 quart leftover heated the next day for six minutes in a 1000W microwave reaches 55° C.

I do stretches every day to accommodate a L5-S1 disc replacement with a titanium-cage-and-rods apparatus done ten years ago, and a C5-C6-C7 similar operation done eleven years ago. Can’t say whether recent diet, last decades’ disc replacement surgeries, daily stretches and exercises, or other factors are responsible for absence of spine pain.

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ω-6 to ω-3 PUFA ratio

Three human-evidenced publications on omega-6 and omega-3 polyunsaturated fatty acids, with the first a 2021 blog post that cited 72 references:

“In the area of heart health, which is why most consumers swallow fish oil, the data is hopelessly conflicted:

  • One meta-analyses found that protective effects were dose-related, which is always persuasive;
  • In marked contrast, three recent powerful clinical trials found fish oil to have no effects on cardiovascular pathology in either primary or secondary prevention; and
  • Yet another meta-analysis found null results, except for a slight degree of protection in subjects who had gallantly taken fish oil supplements for over ten years.

Can these all be right? I think they can, based on secondary bioavailability.

Levels of omega 3s in the bloodstream are irrelevant, except in terms of their calorie content. That is not where they do their anti-inflammatory thing. They become precursors for resolvins, maresins, protectins, and anti-inflammatory eicosanoids only after they have been incorporated into the host’s cell membranes.

Getting them into cell membranes is secondary bioavailability (or bio-efficacy), and this is a much more complicated procedure. Seafood does it, but fish oil doesn’t.

Specifically, there is something in oily fish which enables secondary bioavailability, but which is missing in commercial fish oils. That something is a lipophillic polyphenol called phlorotannin.”

https://drpaulclayton.eu/blog/fish-oil-upgrade-to-snake-oil/ “Fish Oil? Upgrade to Snake Oil!”


A second paper was a 2021 review that focused on ratios of ω-6 to ω-3 PUFAs:

“Chronic diseases including obesity, type 2 diabetes, cardiovascular disease, cancer, and Alzheimer’s disease are rising exponentially in the modern world. Though these diseases are multifactorial in nature, their prevalence is mostly associated with an unbalanced increase in dietary n-6 PUFAs and decrease in n-3 PUFAs.

Mostly, these diseases escalate on the fact that inflammation in conjunction with obesity is the basis of every chronic disease.

Considering antagonistic effects of n-3 and n-6 PUFAs, both n-3 and n-6 SC-PUFAs and LC-PUFAs in their proportional ratio with each other, which is close to 4:1, play a significant role in regulating body homeostasis of inflammation and anti-inflammation, vasodilation and vasoconstriction, bronchoconstriction and bronchodilation, and platelet aggregation and antiaggregation.”

https://www.hindawi.com/journals/jl/2021/8848161/ “Overconsumption of Omega-6 Polyunsaturated Fatty Acids (PUFAs) versus Deficiency of Omega-3 PUFAs in Modern-Day Diets: The Disturbing Factor for Their ‘Balanced Antagonistic Metabolic Functions’ in the Human Body”


A third paper was a 2020 human adolescent study:

“Obese youth 9–19 y of age with nonalcoholic fatty liver disease were treated to see whether 12 wk of a low n–6:n–3 PUFA ratio (4:1) normocaloric diet mitigated fatty liver.

Independent of weight loss, a low n–6:n–3 PUFA diet ameliorated the metabolic phenotype of adolescents with fatty liver disease. This trial was registered at clinicaltrials.gov as NCT01556113.”

https://academic.oup.com/jn/article/150/9/2314/5870325 “A Low ω-6 to ω-3 PUFA Ratio (n–6:n–3 PUFA) Diet to Treat Fatty Liver Disease in Obese Youth”


My ω-6 to ω-3 PUFA 4 : 1 (1400 / 350) intake at breakfast and dinner via Balance Oil:

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At lunch I eat an ounce of walnuts with a ω-6 to ω-3 PUFA 4.4 : 1 ratio:

walnuts 1 oz


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A time to speak

“To every thing there is a season, and a time to every purpose under heaven:
A time to break down, and a time to build up;
A time to mourn, and a time to dance;
A time to embrace, and a time to refrain from embracing;
A time to keep silent, and a time to speak.”


A review from 2017:

“Few, if any, other drugs can rival ivermectin for its beneficial impact on human health and welfare. Perhaps more than any other drug, ivermectin is a drug for the world’s poor. For most of this century, some 250 million people have been taking it.

The following are an indication of disease-fighting potential that has been identified for ivermectin thus far:

  • Antiviral – Ivermectin has been found to potently inhibit replication of yellow fever virus, with EC50 values in the sub-nanomolar range. It inhibits replication in several other flaviviruses, including dengue, Japanese encephalitis, and tick-borne encephalitis. Ivermectin interrupts virus replication. It demonstrates antiviral activity against several RNA viruses by blocking nuclear trafficking of viral proteins. It has been shown to have potent antiviral action against HIV-1.
  • Asthma – Ivermectin suppressed mucus hypersecretion by goblet cells, establishing that ivermectin can effectively curb inflammation, such that it may be useful in treating allergic asthma and other inflammatory airway diseases.
  • Bedbugs – Ivermectin is highly effective against bedbugs, capable of eradicating or preventing bedbug infestations.
  • Disease vector control – Ivermectin is highly effective in killing a broad range of insects. Comprehensive testing against 84 species of insects showed that avermectins were toxic to almost all insects tested. At sub-lethal doses, ivermectin inhibits feeding and disrupts mating behavior, oviposition, egg hatching, and development.
  • Malaria – Mosquitoes that transmit Plasmodium falciparum, the most dangerous malaria-causing parasite, can be killed by ivermectin present in the human bloodstream after a standard oral dose.
  • Myiasis – Myiasis is an infestation of fly larvae that grow inside the host. Oral myiasis has been successfully treated with ivermectin, which has also been effective as a non-invasive treatment for orbital myiasis, a rare and preventable ocular morbidity.
  • Schistosomiasis – Schistosoma species are the causative agent of schistosomiasis, a disease afflicting more than 200 million people worldwide. Ivermectin helps control one of the world’s major neglected tropical diseases.
  • Trichinosis – Globally, approximately 11 million individuals are infected with Trichinella roundworms. Ivermectin kills Trichinella spiralis, the species responsible for most of these infections.”

https://www.nature.com/articles/ja201711 “Ivermectin: enigmatic multifaceted ‘wonder’ drug continues to surprise and exceed expectations”


59 citations in CrossRef. Didn’t see citing 2020-2021 papers that noted any safety concerns when administered at proper doses.

Train your immune system every day, because:

“Rapid clearance following ivermectin dosing, results not from direct impact of the drug, but via suppression of a parasite’s ability to evade the host’s natural immune defense mechanisms.”

It’s safe, and it’s effective. Ivermectin’s main difficulty is that its patent expired in 1997.

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Improving gut barriers

Three papers on gut barriers, with the first a 2020 review of four intestinal barrier layers:

“The epithelial cell layer and outer/inner mucin layer constitute the physical barrier. Intestinal alkaline phosphatase (IAP) produced by epithelial cells and antibacterial proteins secreted by Panneth cells represent the functional barrier.

Multiple layers of this barrier, from intestinal lumen to systemic circulation, include:

  1. Luminal intestinal alkaline phosphatase (IAP) that dephosphorylates bacterial endotoxin lipopolysaccharide (LPS) to detoxify it;
  2. Mucus layer that provides a physical barrier preventing interactions between gut bacteria and intestinal epithelial cells;
  3. Tight junctions between epithelial cells that limit paracellular transport of bacteria and/or bacterial products to systemic circulation; and
  4. Antibacterial proteins secreted by specialized intestinal epithelial cells or Paneth cells, and IgA [immunoglobulin A] secreted by immune cells present in lamina propria underlying the epithelial cell layer.

m_bvz039f0001

The presence of LPS in systemic circulation is identified as a causal or complicating factor in diverse diseases such as:

  • Diet-induced metabolic diseases;
  • Autism;
  • Alzheimer’s disease;
  • Parkinson’s disease;
  • Arthritis;
  • Obesity-induced osteoarthritis;
  • Asthma; and
  • Several autoimmune diseases.

Causal relationships between circulating LPS levels and development of multiple diseases underscore the importance of changes in intestinal barrier layers associated with disease development.

Correcting intestinal barrier dysfunction to modulate multiple diseases can be envisioned as a viable therapeutic option. Identifying precise defects by use of specific biomarkers would facilitate targeted interventions.”

https://academic.oup.com/jes/article/4/2/bvz039/5741771 “Intestinal Barrier Dysfunction, LPS Translocation, and Disease Development”


A second 2020 review focused on IAP:

“IAP plays a vital role in intestinal barrier function, affecting bicarbonate secretion, duodenal surface pH, nutrient resorption, local intestinal inflammation, and gut microbiota. Disturbances of IAP functions are associated with persistent inflammatory diseases associated with aging (i.e.,inflammageing), inflammatory bowel diseases, type 2 diabetes mellitus, obesity, metabolic syndrome, and chronic kidney disease (CKD).

Expression and activity of IAP are directly affected by food intake, i.e., quantity and type of macro- and micronutrients including vitamins and other bioactive nutrients, or by absence of food, as well as indirectly by composition of gut microbiota that in turn are highly dependent on food intake. Increased IAP gene expression and activity promoting detoxification of LPS may lead to improvement of both intestinal and systemic inflammation, reduced bacteria translocation, and maintaining gut barrier function.

IAP could be used as an inflammatory marker together with other markers, such as interleukins, to predict inflammation and diseases that are based on chronic inflammatory processes.”

https://doi.org/10.1007/s13167-020-00228-9 “Intestinal alkaline phosphatase modulation by food components: predictive, preventive, and personalized strategies for novel treatment options in chronic kidney disease” (not freely available)


A third paper was a 2021 rodent study by coauthors of the first paper:

“We developed intestine-specific IAP transgenic mice (IAPTg) overexpressing human chimeric IAP to examine direct effects of increased IAP expression on barrier function and development of metabolic diseases. We evaluated effects of intestine-specific IAP overexpression in hyperlipidemic Ldlr−/− mice. The data presented demonstrated significant attenuation of Western-type diet (WD)-induced LPS translocation in Ldlr−/−IAPTg mice, with significant reduction in intestinal lipid absorption, hyperlipidemia, hepatic lipids, and development of atherosclerotic lesions.

circresaha.120.317144.fig09

IAP is produced by enterocytes, and catalyzes removal of 1 of the 2 phosphate groups from the toxic lipid A moiety of LPS. This produces monophosphoryl-LPS, and results in attenuation of the downstream TLR (Toll-like receptor)-4–dependent inflammatory cascade.

IAP also:

  • Dephosphorylates other proinflammatory molecules such as flagellin and ATP, resulting in their detoxification;
  • Regulates expression of key gap junction proteins (zonula occludens, claudin, and occludin) and their cellular localization, which directly modulates intestinal barrier function;
  • Promotes growth of various commensal bacteria in the gut by decreasing luminal concentrations of nucleotide triphosphates via dephosphorylation; and
  • Translocates from the apical surface of enterocytes during fat absorption. Increased serum IAP accompanies fat absorption, which is consistent with observed increased levels of circulating LPS in WD-fed mice, providing one more likely mechanism by which WD affects intestinal barrier function via IAP.

Nutrients and food components/supplements that increase IAP include galacto- or chito- oligosaccharides, glucomannan, and vitamin D3. These provide a novel opportunity to develop simple strategies for modulation of diet/nutrition to target metabolic diseases including diabetes, fatty liver disease, atherosclerosis, and heart disease.”

https://www.ahajournals.org/doi/10.1161/CIRCRESAHA.120.317144 “Over-Expression of Intestinal Alkaline Phosphatase Attenuates Atherosclerosis”


Previously curated IAP studies were:

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Berry polyphenols

This 2021 review subject was berries and health:

“Phenolic compounds present in different berries (raspberry, blueberry, goji berry, black currant, strawberry, cranberry, and blackberry) were summarized based on up-to-date information and their beneficial health effects. Compounds such as anthocyanins, flavonols, and phenolic acids occur in different concentrations depending on berry type.

1-s2.0-S2214799321001028-fx1_lrg

Polyphenols are the ‘new’ prebiotics. A more recent definition of prebiotics is ‘a substrate that is selectively utilised by host microorganisms and conferring a health benefit.’

Only 5–10% of total intake is absorbed in the small intestine. Remainders can reach the large intestinal lumen, where they may be subjected to gut microbial community enzymatic activities. Microbiota can catabolize flavonoids that have not been absorbed into smaller molecules, such as phenolic and aromatic acids, which can then be absorbed by intestinal villi.

Increase of beneficial bacteria such as Bacteroidetes, decrease of Firmicutes, and production of short-chain fatty acids is almost consensus among studies. More in vivo data are required to understand mechanisms of action, while clinical trials using different characteristics (i.e., gender, age, existence of diseases) should be performed so new information on bioactivity of berries can be unveiled.”

https://www.sciencedirect.com/science/article/abs/pii/S2214799321001028 “Berry polyphenols and human health: evidence of antioxidant, anti-inflammatory, microbiota modulation, and cell-protecting effects (not freely available) Thanks to Dr. Anderson S. Sant’Ana for providing a copy.


It’s summer, and time to gorge on berries! We’ll deal with overindulgences later.

IMG_20190705_100825

Gut and brain health

This 2021 human review subject was interactions of gut health and disease with brain health and disease:

“Actions of microbial metabolites are key for appropriate gut-brain communication in humans. Among these metabolites, short-chain fatty acids (SCFAs), tryptophan, and bile acid metabolites / pathways show strong preclinical evidence for involvement in various aspects of brain function and behaviour.

Dietary fibres, proteins, and fats ingested by the host contain components which are metabolized by microbiota. SCFAs are produced from fermentation of fibres, and tryptophan-kynurenine (TRP-KYN) metabolites from dietary proteins. Primary bile acids derived from liver metabolism aid in lipid digestion, but can be deconjugated and bio-transformed into secondary bile acids.

1-s2.0-S0149763421001032-gr1

One of the greatest challenges with human microbiota studies is making inferences about composition of colonic microbiota from faeces. There are known differences between faecal and caecal microbiota composition in humans along with spatial variation across the gastrointestinal tract.

It is difficult to interpret microbiome-host associations without identifying the driving influence in such an interaction. Large cohort studies may require thousands of participants on order to reach 20 % explanatory power for a certain host-trait with specific microbiota-associated metrics (Shannon diversity, relative microbial abundance). Collection of metadata is important to allow for a better comparison between studies, and to identify differentially abundant microbes arising from confounding variables.”

https://www.sciencedirect.com/science/article/pii/S0149763421001032 “Mining Microbes for Mental Health: Determining the Role of Microbial Metabolic Pathways in Human Brain Health and Disease”


Don’t understand why these researchers handcuffed themselves by only using PubMed searches. For example, two papers were cited for:

“Conjugated and unconjugated bile acids, as well as taurine or glycine alone, are potential neuroactive ligands in humans.”

Compare scientific coverage of PubMed with Scopus:

  • 2017 paper: PubMed citations 39; Scopus citations 69.
  • 2019 paper: PubMed citations 69; Scopus citations 102.

Large numbers of papers intentionally missing from PubMed probably influenced this review’s findings, such as:

  1. “There are too few fibromyalgia and migraine microbiome-related studies to make definitive conclusions. However, one fibromyalgia study found altered microbial species associated with SCFA and tryptophan metabolism, as well as changes in serum levels of SCFAs. Similarly, the sole migraine-microbiota study reported an increased abundance of the kynurenine synthesis GBM (gut-brain module).
  2. Due to heterogeneity of stroke and vascular disease conditions, it is difficult to make substantial comparisons between studies. There is convincing evidence for involvement of specific microbial genera / species and a neurovascular condition in humans. However, taxa were linked to LPS biosynthesis rather than SCFA production.
  3. Several studies suggest lasting microbial changes in response to prenatal or postnatal stress, though these do not provide evidence for involvement of SCFA, tryptophan, or bile-acid modifying bacteria. Similar to stress, there are very few studies assessing impact of post-traumatic stress disorder on microbiota.”

These researchers took on a difficult task. Their study design could have been better.


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Wildlife

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Vitamin K-dependent proteins

This 2020 review focused on three Vitamin K-dependent proteins (VKDPs):

“We summarize three important emerging VKDPs: Growth arrest‑specific protein 6 (Gas 6), Gla‑rich protein (GRP) and periostin in terms of their functions in physiological and pathological conditions. As examples:

  • Carboxylated Gas 6 and GRP effectively protect blood vessels from calcification;
  • Gas 6 protects from acute kidney injury and is involved in chronic kidney disease;
  • GRP contributes to bone homeostasis and delays progression of osteoarthritis; and
  • Periostin is involved in all phases of fracture healing and assists myocardial regeneration in the early stages of myocardial infarction.

IJMM-47-03-4835-g00

The ‘+’ refers to promotion and ‘-‘ refers to inhibition. Green represents Gas 6 physiological effects and red represents its pathological effects.

  • Gas 6 resists vascular calcification: i) Gas 6 promotes proliferation and migration of endothelial progenitor cells (EPCs); ii) Gas 6 inhibits apoptosis and senescence of vascular smooth muscle cells (VSMCs) by binding Tyro3, Axl and Mer (TAM) receptors; iii) Gas 6 decreases expression of inflammatory factors, including TNF-α and ICAM-1.
  • Gas 6 protects from acute kidney injury: i) Gas 6 significantly reduces creatinine and blood urea nitrogen; ii) Gas 6 enhances macrophages to uptake apoptotic cells; iii) Gas 6 reduces the expression of pro-inflammatory cytokines, such as IL-1β.
  • Gas 6 assists tumor progression: i) Gas 6 is necessary for survival, proliferation and growth of tumor cells; ii) Gas 6 contributes to drug resistance and tumor angiogenesis; iii) Gas 6 negatively regulates tumor immunity.

Numerous physiological benefits of vitamin K2 have been identified, such as anti-vascular calcification, glycemic control, and lipid-lowering effects. However, some questions about relationships between vitamin K2 and cancers remain unsolved. VKDPs are expected to be biomarkers for many diseases.”

https://www.spandidos-publications.com/10.3892/ijmm.2020.4835?text=fulltext “Role of emerging vitamin K‑dependent proteins: Growth arrest‑specific protein 6, Gla‑rich protein and periostin (Review)”


This review’s VKPD biomarkers included:

  • Vascular calcification;
  • Asthma;
  • Bronchial obstruction;
  • Diabetic nephropathy; and
  • Fracture risk.

Elaborating on this last item:

“In a cohort of 607 postmenopausal women from France that were followed up for 7 years, a positive correlation between serum periostin and fracture risk was observed. The association was independent of bone mineral density and prior fractures, indicating that periostin is an independent predictive marker of fracture risk.”

As pointed out in Chronological age by itself is an outdated clinical measurement, bone mineral density is one of several historical measurements that were selected for their relative convenience instead of chosen for their efficacy. We’re in a different century now.

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Part 2 of Week 63 of Changing to a youthful phenotype with sprouts

To follow up Part 1, received Thursday’s lab results yesterday. Downloaded the workbook at https://michaellustgarten.com/2019/09/09/quantifying-biological-age/ and filled it in. Went to http://aging.ai/, selected 3.0, and entered values.

My starting point’s calculated values were:

biological age 1

A biological age snapshot from a year ago‘s video included optimal ranges:

optimizing biological age

Values in these optimal ranges were:

  • Albumin: 46;
  • Creatinine: 1.07;
  • high-sensitivity C-reactive protein: 0.24;
  • Red cell distribution width: 11.8; and
  • White blood cell count: 4.6.

I have some work to do on the other four. Good health while aging seldom happens on its own.

Reading more about Phenotypic age and its biological relationships. It definitely doesn’t mean I can do things I did 9.5 years ago like play golf and Frisbee football on the weekends.

I’d probably use DNAm PhenoAge to compare with other epigenetic clocks. Not sure how to use Aging.ai 3.0 calculations.

Sometime over the past year, Labcorp changed their adult alkaline phosphatase reference range from 39-117 to 48-121. Don’t know whether alkaline phosphatase’s optimal range will change with Labcorp’s new range, since < 48 was based on all-cause-mortality data.

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Eat whole oats for your gut microbiota

Two papers on whole grains, with the first a 2021 review:

“Whole grains are more complex than refined grains and are promoted as part of a healthy and sustainable diet, mainly because the contribution of indigestible carbohydrates, and their co-passenger nutrients, is significantly higher. Changing composition and availability of grain carbohydrates and phytochemicals during processing may positively affect gut microbiota and improve health.

Processing is required for virtually all cereals that humans consume. However, eliminating bran has resulted in grain-based products that contribute to a lower-quality diet.

Currently, there are no specific recommendations on relative proportions of different dietary fiber types (based on variability in fermentability or degree of solubility). Switching from refined grain to whole grain will deliver more dietary fiber and nutrients associated with bran and germ, and improve diet quality.

crf312728-fig-0001-m

Carbohydrate-rich foods that are higher in slowly digested starches, resistant starch, oligosaccharides with prebiotic potential, and dietary fiber are considered to have a higher quality. Foods can be awarded an overall carbohydrate quality index (CQI). The optimum ratio of total carbohydrate (CHO) to dietary fiber should be ≤10:1.

Mostly only oligosaccharides and polysaccharides reach the colon. Even though larger molecules were fermented slower, they were still fermented within the proximal colon.

It is not surprising that there are conflicting reports with respect to effects of whole grains on gut microbiota. Part of this is due to whole grains comprising a diverse group of staple cereal foods, including wheat, corn, rice, oats, barley and rye, and hence different effects on gut microbiota are expected. Differences in study design, with respect to dose, duration, and study populations make it difficult to compare between studies and distill overarching similarities.

Enzymes can modify less fermentable dietary fiber to improve its fermentability by microbiota. Using different enzymes, dietary fibers can contribute to fermentation throughout the colon.”

https://onlinelibrary.wiley.com/doi/10.1111/1541-4337.12728 “Health benefits of whole grain: effects on dietary carbohydrate quality, the gut microbiome, and consequences of processing”


This review cited a 2019 paper as “an elegant study where oat bran (including co-passengers) was shown to be effective in increasing Bifidobacterium populations in the gut, whereas purified bioactive β-glucans did not show a bifidogenic effect”:

“Whole grain oats are known to modulate human gut microbiota and have prebiotic properties. Research todate mainly attributes these effects to fibre content. However, oats are also a rich dietary source of polyphenols, which may contribute to positive modulation of gut microbiota.

We found that oats increased bifidobacteria, acetic acid and propionic acid. This was mediated by synergy of all oat compounds within the complex food matrix, rather than its main bioactive β-glucan or polyphenols.

While human digestive enzymes cannot degrade plant cell wall polysaccharides, gut xylanolytic bacteria can, producing SCFA with health-beneficial effects. Certain strains down-regulate gene and protein expression of pro-inflammatory cytokines, notably isoform of nitric oxide synthase and PPAR-γ and interferon-γ, resulting in reduced inflammatory status, suggesting that oat β-glucan have beneficial effects on human health.

Oats as a whole food led to the greatest impact on microbiota.”

https://www.cambridge.org/core/journals/british-journal-of-nutrition/article/oat-bran-but-not-its-isolated-bioactive-glucans-or-polyphenols-have-a-bifidogenic-effect-in-an-in-vitro-fermentation-model-of-the-gut-microbiota/B23FAE2C7EED702132FC72F1C9CE990E “Oat bran, but not its isolated bioactive β-glucans or polyphenols, have a bifidogenic effect in an in vitro fermentation model of the gut microbiota”


The Avena nuda oats I eat for breakfast start out as 81.0 grams (1/2 cup). The only processing I do from an Illinois farmer is soaking them for 16 hours, draining then changing out to 1 1/2 cups water, then cooking for 20 minutes in a 1000W microwave at 80% power. They end up weighing 154.7 g.

I eat 51.9 g of 3-day-old sprouted Avena sativa oats from a Montana farmer at the same time, and concurrently take 2.5 g inulin. Pretty sure this 154.7 + 51.9 + 2.5 = 209.1 g combination meets an “optimum ratio of total carbohydrate to dietary fiber ≤10:1.”

Also pretty sure sprouted Avena sativa oats supply enzymes that facilitate breaking down Avena nuda complex molecules. Haven’t experienced any complaints over the past 3+ months. 🙂

Go for the win-win with taurine

This 2021 rodent study investigated amino acid taurine effects on colitis:

“Taurine plays an important role in various essential biological processes. Health beneficial effects of taurine have been generally attributable to its antioxidant and anti-inflammatory effects.

Taurine chloramine (TauCl) is an endogenous anti-inflammatory substance derived from taurine. In fighting exogenous pathogens, neutrophils utilize one powerful weapon in their arsenal: generating the strong oxidant hypochlorous acid (HOCl), which is nature’s germ killer.

Taurine can act as a trap for HOCl forming the long-lived oxidant TauCl, which is more stable and less toxic than HOCl. TauCl (20 mg/kg) was given on daily basis by gavage for 10 days before and for 3 days after intrarectal administration of 2.5% TNBS:

antioxidants-10-00479-g008-550

TauCl inhibits generation of proinflammatory mediators by phagocytic cells. Taurine exerts an anti-inflammatory as well as antioxidant action by preventing cytolytic damage caused by HOCl generated by inflammatory cells, particularly neutrophils.

These results suggest that TauCl exerts a protective effect against colitis through upregulation of Nrf2-dependent cytoprotective gene expression, while blocking proinflammatory signaling mediated by NFκB and STAT3.”

https://www.mdpi.com/2076-3921/10/3/479/htm “Protective Effects of Taurine Chloramine on Experimentally Induced Colitis: NFκB, STAT3, and Nrf2 as Potential Targets”


Other curated taurine studies include:

Broccoli sprouts positively influence Sestrin proteins

Four papers on Sestrin, with the first a 2021 review:

“Sestrin 2 (Sesn2) is a member of the evolutionarily conserved and stress-inducible sestrin family. In mammals, this family is composed of Sesn1–3, and Sesn2 is the main member that responds to oxidative stress.

Sesn2 inhibits mammalian target of rapamycin (mTOR)-mediated cell over-proliferation by activating adenosine monophosphate-activated protein kinase (AMPK) and its kinase activity. Sesn2 also regulates redox balance by directly exerting antioxidant enzyme activity and regulating antioxidant signaling.

Inflammation, which is not regulated by oxidative stress, also plays an important role in cardiovascular diseases (CVDs). Sesn2 is involved in inflammation and immune regulation in many systems.

There is a positive feedback loop between Sesn2 and Nrf2:

sestrin2 nrf2

Sesn2 and p62 are expressed under oxidative stress. Sesn2 binds to ULK1 and p62 to form a functional complex, which promotes p62 phosphorylation, promoting p62-dependent autophagy degradation of Keap1.

Consequently, Nrf2 accumulates in cells, transfers to the nucleus, and promotes transcriptional activation of genes controlled by antioxidant response elements (ARE).

Circulating Sesn2 levels are elevated in a variety of CVDs, such as coronary heart disease, heart failure and atrial fibrillation, which indicates that Sesn2 is induced and plays a protective role in CVDs.”

https://www.sciencedirect.com/science/article/abs/pii/S0891584920316270 “Sestrin 2, a potential star of antioxidant stress in cardiovascular diseases” (not freely available)


A second paper was also a 2021 review:

“Sestrin2 acts as an antioxidant protein that diminishes accumulation of ROS and inhibits mTORC1 signaling. Both accumulation of ROS and activation of mTORC1 are associated with aging and age-related diseases.

Since plasma sestrin2 levels in patients with CAD and those with carotid atherosclerosis were shown to be high, it remains unclear whether or not an exogenous administration of sestrin2 could be beneficial for prevention of atherosclerotic disease.”

https://www.mdpi.com/1422-0067/22/3/1200/htm “The Protective Role of Sestrin2 in Atherosclerotic and Cardiac Diseases”


A third paper was a 2020 human study:

“Sesn 1 and Sesn 2 levels were significantly reduced in sarcopenic compared to non-sarcopenic subjects. It can be concluded that sarcopenia can be diagnosed at the early stage by using serum sestrin as one potential biomarker.”

https://link.springer.com/article/10.1007/s40520-020-01642-9 “Serum sestrins: potential predictive molecule in human sarcopenia” (not freely available)


A fourth paper was a 2020 rodent study:

“Sulforaphane (SFN) alleviated hematological variations, oxidative stress, heart dysfunction and structure disorder, and cardiomyocyte apoptosis induced by potassium dichromate. Moreover, SFN:

  • Reduced p53;
  • Cleaved caspase-3, Bcl2-associated X protein, nuclear factor kappa-B, and interleukin-1β levels; and
  • Increased Sesn2, Nrf2, heme oxygenase-1, NAD(P)H quinone oxidoreductase-1; and
  • Phosphorylated AMPK levels.

This study demonstrated that SFN ameliorates Cr(VI)-induced cardiotoxicity via activation of the Sesn2/AMPK/Nrf2 signaling pathway.”

https://pubs.rsc.org/en/content/articlelanding/2020/mt/d0mt00124 “Sulforaphane attenuates hexavalent chromium-induced cardiotoxicity via the activation of the Sesn2/AMPK/Nrf2 signaling pathway” (not freely available)


I found these studies as well as the previous post Cow milk causes disease from their citing a 2015 study The antioxidant function of sestrins is mediated by promotion of autophagic degradation of Keap1 and Nrf2 activation and by inhibition of mTORC1 (not freely available).

Cow milk causes disease

This 2021 review followed up Epigenetic effects of cow’s milk and many papers since then:

“Epidemiological studies associate intake of cow milk with an increased risk of diseases, which are associated with overactivated mechanistic target of rapamycin complex 1 (mTORC1) signaling. Milk’s physiological function to maintain high mTORC1 signaling at the beginning of mammalian life turns into adverse health effects when this postnatal endocrine and epigenetic system is not discontinued as designated by physiological processing of the lactation genome.

Milk is a signaling interface between the maternal lactation genome and the infant’s cellular mTORC1 system that orchestrates growth, anabolism, metabolic, immunological, and neurological programming. Pasteurization combined with refrigeration exposed human milk consumers to bioactive milk exosome (MEX)-derived micro-ribonucleic acids (miRs), augmenting milk’s mTORC1 activity compared to boiled, ultra-heat-treated, or fermented milk.

milk-mediated mTORC1 signaling

Milk consumption activates five major pathways stimulating mTORC1 via:

  1. Growth factors, including growth hormone, insulin, and insulin-like growth factor 1;
  2. Amino acids, especially branched-chain amino acids;
  3. Milk fat-derived palmitic acid;
  4. Milk sugar lactose; and
  5. Epigenetic modifiers, especially MEX-derived miRs.

Understanding milk’s interaction with the central hub of metabolic regulation, mTORC1, will open new avenues for prevention of common diseases.”

https://www.mdpi.com/2218-273X/11/3/404/htm “Lifetime Impact of Cow’s Milk on Overactivation of mTORC1: From Fetal to Childhood Overgrowth, Acne, Diabetes, Cancers, and Neurodegeneration”


This reviewer is somewhat of a zealot. Still, he cited 555 references.

His genotype may tolerate lactose, but he didn’t argue for it:

“After breast feeding, mucosal expression of lactase, an intestinal enzyme hydrolyzing lactose into glucose and galactose, is downregulated in all mammals with the exception of Neolithic humans, who developed LCT [lactase gene] mutations allowing persistent lactase expression.

Lactose content of milk makes up around 2–8% by weight. Lactose hydrolysis provides glucose and galactose, which both activate mTORC1:

  • During glucose abundance and glycolysis, sufficient cellular energy is produced in the form of ATP, which suppresses AMPK activity. Aldolase operates as a sensor for glucose availability that directly links glucose shortage to activation of AMPK.
  • Galactose via induction of oxidative stress activates mTORC1. Galactose-induced overactivation of mTORC1 promotes senescence of neural stem cells and aging of mesenchymal stem cells.

Lactobacilli used in food and dairy fermentation increase NRF2 activation, resulting in NRF2-induced sestrin expression, which attenuates mTORC1 activation.”

Eat broccoli sprouts for your hearing

Two 2021 papers, both of which I found by each citing a 2009 Molecular mechanisms underlying cochlear degeneration in the tubby mouse and the therapeutic effect of sulforaphane (not freely available). First was a review:

“Hair cell damage and loss mediated by oxidative stress are important causes of hearing loss. Sensorineural hearing loss is the most common type of hearing loss, including noise induced hearing loss (NIHL), age-related hearing loss (ARHL), and ototoxic hearing loss.

Nrf2 reduces cell damage caused by oxidative stress, and maintains the dynamic balance of systematic redox by inducing and regulating expression of various antioxidant factors. This review summarizes correlation studies of Nrf2 in hearing loss, providing ideas for prevention and treatment of hearing loss with Nrf2 as the target.

fphar-12-620921-g002

There is positive feedback between p62-mediated autophagy and Nrf2. p62 promotes accumulation of Nrf2 and nuclear translocation. Concurrently, increased Nrf2 promotes p62 expression.

How Nrf2 regulates ROS changes in hair cells, and the upstream and downstream regulatory network of Nrf2 in hair cells, are still not fully understood. Studies on early prevention and treatment of hearing loss through the Keap1-Nrf2-ARE [antioxidant response element] signaling axis are still at the exploratory stage.”

https://www.frontiersin.org/articles/10.3389/fphar.2021.620921/full “The Role of Nrf2 in Hearing Loss”


Second paper was a rodent study:

“We examined oxidative stress and antioxidant response of the p62-Keap1-Nrf2 pathway in cochleae during age-related hearing loss (ARHL) and noise-induced hearing loss (NIHL). We elucidated the function of full-length and variant p62/Sqstm1 (referred to here as p62) in regulation of Nrf2 activation.

Cochlear damage was assessed by analyzing auditory brainstem response (ABR) and by counting hair cells (HCs). Malondialdehyde (MDA, a lipid peroxidation product) levels were measured in young and old mice to determine whether oxidative stress contributed to ARHL.

auditory brainstem response

  • (A) Audiometric threshold (dB) determined from click and pure tone evoked ABRs. Thresholds were each significantly different (P < 0.001) between young mice and old mice.
  • (B) HC loss percentage in basal cochlear turns. Significant differences (P < 0.001) were observed between young and old mice.
  • (C) MDA levels in the cochleae of old mice were significantly higher (P = 0.034) than those of young mice.

ROS accumulation is closely related to ARHL and NIHL. The inability of ROS accumulation to activate the Nrf2 antioxidant stress pathway under physiological conditions may be related to alternative splicing of p62 mRNA in cochleae.

However, the agonist of the Nrf2 pathway enhanced Nrf2 nuclear translocation. This suggests a mechanism in which the antioxidant pathway was difficult to be activated in the context of accumulation of ROS.”

https://www.researchsquare.com/article/rs-535219/v1 “New Target of Oxidative Stress Regulation in Cochleae:Alternative Splicing of the p62/Sqstm1 gene”


The study’s two-month-old mice were equivalent to a 20-year-old human. Its 13-to-14-month-old mice were equivalent to humans in their 60s to 70s.

I expected preconditioning to be mentioned in both papers. Maybe these researchers thought it was too obvious and didn’t need to be stated that:

  • Repeated use of a Nrf2 activator produces transient mild stress;
  • Which elicits a stronger response; and
  • Preconditions cells for future stress?

Sulforaphane in the Goldilocks zone and its cited papers exhaustively emphasized preconditioning’s importance. The main thing I’m trying to do with isothiocyanates is to send a weak pro-inflammatory signal to my endogenous ARE system to exercise natural defenses.

Twice-daily drills make me more proficient at responding to actual emergencies. Post-drill, my body recycles material to be ready to respond the next time.

I do the same thing once a day with β-glucan 1,3/1,6 to train my innate immune system. Microphages in my gut are the first responders. Like the very reactive isothiocyanates, I don’t take anything with, or an hour before or after β-glucan 1,3/1,6.

Why tolerate “the antioxidant pathway was difficult to be activated in the context of accumulation of ROS” when a sulforaphane “agonist of the Nrf2 pathway enhanced Nrf2 nuclear translocation”? For all we know, diminished natural defenses and hearing loss may exist to turn old mammals into prey.

Continued in Part 2.

Part 3 of Broccoli sprouts activate the AMPK pathway

This 2020 cell study investigated sulforaphane and three transcription pathways:

“Novel findings of this study are:

  1. AMPK controls only a subset within the Nrf2-dependent transcriptome;
  2. Altered Nrf2 levels or altered accessibility of regulatory ARE sites do not account for observed differences in target gene transcription between used wt and AMPK −/− cells;
  3. Rather, AMPK presence/activity ensures reduced Bach1 abundance with preferential Nrf2 over Bach1 binding to regulatory ARE sites, and finally stronger transactivation of selected target genes; and
  4. AMPK negatively controls bach1 mRNA expression.

fcell-08-00628-g009

In AMPK−/− cells, levels of BTB and CNC homology 1 (Bach1), a competitor of Nrf2 for ARE sites with predominant repressor function, were higher. Bach1 also bound to a greater relative extent to the examined ARE sites when compared to Nrf2.

Observed AMPK-mediated boost in transactivation of a subset of Nrf2 target genes involves downregulation of Bach1 and subsequent favored binding of activating Nrf2 over repressing Bach1 to examined ARE sites.

fcell-08-00628-g001

The discovered link between AMPK and Bach1 as well as the resulting selective influence on Nrf2 target gene expression are compelling and touch existing data:

  • Bach1 contributed to expression of only selected Nrf2 target genes in endothelial cells under hypoxic conditions which, in turn, are known to influence AMPK activity.
  • Bach1 levels are elevated during aging, in metastatic lung tumors or triple negative breast tumors with concomitant mitochondrial dysfunction, all events also partly connected with AMPK- and/or Nrf2 activity.

These issues strongly advocate for a closer look into interplay between cellular sensors and executors of the oxidative/xenobiotic and metabolic stress response, which likely will uncover additional layers of regulation of cellular stress resilience.”

https://www.frontiersin.org/articles/10.3389/fcell.2020.00628/full “AMPK Enhances Transcription of Selected Nrf2 Target Genes via Negative Regulation of Bach1”


This study hasn’t been cited even once since it was published eleven months ago. These researchers did a very good job of producing evidence for mechanisms of signaling pathways competing with and complementing each other.

This study provided further details to support Broccoli sprouts activate the AMPK pathway findings that sulforaphane first activates the AMPK pathway on the way to its main effect of Nrf2 pathway activation:

figure 8

Brown your white fat cells with broccoli sprouts

A 2021 rodent study and a blog post with 51 references investigated fat cells:

“Sulforaphane (SFN) is a potent indirect antioxidant and a promising agent for controlling metabolic disorder disease. We evaluated efficacy of SFN against high fat diet (HFD)-induced-obesity mice, and investigated potential mechanisms.

SFN:

  • Suppressed HFD-induced body weight gain;
  • Reduced fat cell [adipocyte] size;
  • Suppressed expression of key genes in adipogenesis;
  • Inhibited lipid accumulation in C3H10T1/2 [pre-adipocyte] cells;
  • Increased expression of brown adipocyte-specific markers and mitochondrial biogenesis in vivo and in vitro; and
  • Decreased cellular and mitochondrial oxidative stress.

sulforaphane influences fat cells

Gene expression profile of C3H10T1/2 cells after SFN treatment showed that SFN inhibited expression of core adipogenesis genes (Ppar-γ, Fas, Cebpβ and Scd1) and enhanced expression of browning genes (Chop, Temem 26, Ucp1, Pgc-1α, and Prdm16) in adipocyte differentiation and trans-differentiation. This result suggested possible conversion of white adipocytes into beige cells.

We report that SFN induces browning of mature C3H10T1/2 adipocytes based on promotion of mitochondrial biogenesis by means of upregulation of the AMPK and NRF2 signaling pathways, and enhancement of mitochondrial function. Our further research revealed that SFN can prevent HFD-induced obesity in C57BL/6N mice by inducing browning of white adipose tissue.”

https://www.frontiersin.org/articles/10.3389/fphar.2021.665894/full “The Protective Effects of Sulforaphane on High-Fat Diet-Induced Obesity in Mice Through Browning of White Fat”


Dr. Paul Clayton had a nuanced view of body fat and its browning:

“You can divide adipose tissue into three cell types:

  • White adipocytes account for 95% of all adipocytes and have a primarily storage function;
  • The primary function of brown adipocytes, which range from 1-5% depending on cold exposure and very specific types of chemo-stimulation i.e. β3-adrenergic, is generation of heat via mitochondrial uncoupling.
  • Beige adipocytes are intermediate. They aren’t interspersed in depots of white adipose tissue and can transform into brown-like adipocytes following cold exposure or adrenergic stimulation.

Bone marrow adipose tissue plays an important role in haematopoiesis and bone metabolism in more than one form:

  • One is located in distal bones (forearm and lower leg) and is pretty much stable;
  • The other form is in spine and proximal limb bones, and is inducible by environmental factors such as cold exposure, fasting, and anaemia.

White adipose tissue can be divided into visceral and sub-cutaneous deposits, and these tissues have different behaviours and functions, too.

From a clinical perspective, it’s important to know that adipocyte-related inflammatory effects can be neutralised with omega 3 fatty acids, which return fat cells to a ‘healthy’ configuration. Their inflammatory effects can also be inhibited by various polyphenols which, among other things, block release of pro-inflammatory microRNAs.

In my experience, combining omega 3s with lipophile polyphenols and AMPK-activators such as dammarane saponins and metformin, provide supra-additive benefits.”

https://drpaulclayton.eu/blog/turn-fat-into-muscle/ “Turn Fat into Muscle”


Still no mention of sulforaphane on the doctor’s blog, although it’s:

I came across this first study through a “PPAR sulforaphane” search. Discarding a supplement as a result, because I’m already doing enough!

PXL_20210606_095305180