Broccoli sprout compounds at different growth stages

This 2022 study investigated 12 glucosinolate compounds in 9 broccoli cultivars across seeds, 3-, 11-, and 17-day-old sprouts:

“Broccoli is rich in glucosinolates (GLs) which makes it an excellent source of these nutraceuticals. Composition and concentration of GLs vary among broccoli cultivars and throughout developmental stages of the plant.

9 aliphatic GLs and 3 indole GLs were identified from 9 broccoli cultivars. Aliphatic GLs concentrations decreased with broccoli sprouts and seedling growth for most cultivars. Indole GLs amounts increased after germination and reached the highest level in Stage B 3-day sprouts or C 11-day seedlings, and fell back to a low level in D 17-day seedlings.

Stage B was a stage that sprouts grew with no lights in medium and were about to be transplanted into pots. Stage C was a period that seedlings began to grow a main leaf, while in Stage D they were growing the second main leaf.

stages

Relatively high accumulation of glucoraphanin and glucoerucin in Chunqiujiali seeds suggests that CQJL broccoli seeds could be used for extraction of beneficial aliphatic GLs in nutraceutical industry.”

https://www.mdpi.com/2223-7747/11/12/1563/htm “Variation in Glucosinolate Accumulation among Different Sprout and Seedling Stages of Broccoli (Brassica oleracea var. italica)”


These researchers are probably early in their careers. They may have otherwise measured broccoli sprout compounds like glucosinolate-hydrosolate isothiocyanates and others, as did the cited 3-day-old broccoli sprouts have the optimal yields. As those researchers said:

“From the perspective of comparison methods, broccoli varieties, and germination processes, there is still lack of a systematic comparison of SF yields and other bioactive compounds contents between broccoli seeds and sprouts.”

Glucoraphanin is not sulforaphane highlighted one pitfall in concluding “CQJL broccoli seeds could be used for extraction of beneficial aliphatic GLs in nutraceutical industry.” Selecting broccoli varieties highlighted another, and provided an example of how human-applicable broccoli sprout compound research across different varieties could be done:

“We found a clear difference in selecting functional broccoli by considering only the GSL content or hydrolysates.

  • Even if total GSL content and individual GSL content were high, ITC content could not be produced at a high level.
  • When GSL content is high, if nitrile formation rate was also high, more nitrile than ITC would be produced.”

Preteen practicing handstands 15 minutes before sunrise

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Week 127 of Changing to a youthful phenotype with sprouts

1. My third gut microbiome test results came in this week. I submitted a sample earlier this month to follow methods in the second paper of Improving dietary fiber research in a continuing effort to treat my gut microbiota well.

But that study’s vendor was unable to ship an EU-approved product from The Netherlands to the US because it wasn’t FDA-approved. Our US pets can eat dried chicory root products every day, but we can’t? I haven’t received any positive responses from US vendors of dried chicory root products, so I’ll keep taking up to 10 grams of EU-manufactured inulin daily.

I also followed Dr. Horvath’s suggestion in Epigenetic clocks so far in 2022 to “measure epigenetic age because there’s always an opportunity to make a discovery” and submitted a blood test. Will link to those results when they arrive.

2. These gut microbiome test results highlight a 16S ribosomal RNA technology flaw that Resistant starch therapy pointed out:

“Relative abundances of smaller keystone communities (e.g. primary degraders) may increase, but appear to decrease simply because cross-feeders increase in relative abundance to a greater extent.”

Here are my top two relative abundance results, genus Faecalibacterium and genus Bacteroides:

relative abundance2

25.330% (46,844 total count) of my gut microbiota being a butyrate producer is relatively higher than 22.567% (42,156 total count) 14 months ago. Here’s a review of butyrate’s effects.

25% cross-feeder genus Faecalibacterium didn’t relatively crowd out a primary degrader, genus Ruminococcus, which comparatively stayed at 6%. It may have relatively reduced secondary degrader genus Eubacterium abundance from 6% to 5%.

I don’t assign importance per the above graphic that other people achieve 12% relative abundance of a butyrate producer but I have 25%. Our 10,000+ microbiota species perform many overlapping functions.

Conversely, why should I care that other people host an average 25% genus Bacteroides and I relatively have 17% as I did 14 months ago? It’s similar to irrelevant comparisons of clinical biomarkers in Week 120 of Changing to a youthful phenotype with sprouts.

3. So what are appropriate gut microbiome measurements? They aren’t fine-grained relative measurements of my current gut microbiome, either vs. my previous measurements or vs. other people.

I could make a p < .05 finding out of 25.330% vs. 22.567%. But would those numbers be an adequate proxy for understanding truth?

I think science and industry will affordably catch up to these discrepancies as it has with epigenetic clocks. Haven’t come across well-designed gut microbiota studies that use technologically preferable shotgun metagenomic sequencing with absolute measures of both form and function. I’ve read plenty that are stuck in a relative abundance paradigm.

In the meantime, I’m alright, but have to toughen up quickly so that I can transition later this month from summer weather on my sunrise walk every day to a freezing destination.

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Minds of their own

It’s the weekend, so it’s time for: Running errands? Watching sports? Other conditioned behavior?

Or maybe broadening our cognitive ability with Dr. Michael Levin’s follow-ups to his 2021 Basal cognition paper and 2020 Electroceuticals presentation with a 2022 paper and presentation starting around the 13:30 mark:

Michael Levin - Cell Intelligence in Physiological and Morphological Spaces

“A homeostatic feedback is usually thought of as a single variable such as temperature or pH. The set point has been found to be a large-scale geometry, a descriptor of a complex data structure.”


His 2022 paper Technological Approach to Mind Everywhere: An Experimentally-Grounded Framework for Understanding Diverse Bodies and Minds:

“It is proposed that the traditional problem-solving behavior we see in standard animals in 3D space is just a variant of evolutionarily more ancient capacity to solve problems in metabolic, physiological, transcriptional, and morphogenetic spaces (as one possible sequential timeline along which evolution pivoted some of the same strategies to solve problems in new spaces).

Developmental bioelectricity works alongside other modalities such as gene-regulatory networks, biomechanics, and biochemical systems. Developmental bioelectricity provides a bridge between the early problem-solving of body anatomy and the more recent complexity of behavioral sophistication via brains.

This unification of two disciplines suggests a number of hypotheses about the evolutionary path that pivoted morphogenetic control mechanisms into cognitive capacities of behavior, and sheds light on how Selves arise and expand.

While being very careful with powerful advances, it must also be kept in mind that existing balance was not achieved by optimizing happiness or any other quality commensurate with modern values. It is the result of dynamical systems properties shaped by meanderings of the evolutionary process and the harsh process of selection for survival capacity.”


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Gut microbiota, SCFAs, and hypertension

Two 2022 rodent studies from the same research group on short-chain fatty acid effects, beginning with butyrate:

“Maternal nutrition, gut microbiome composition, and metabolites derived from gut microbiota are closely related to development of hypertension in offspring. A plethora of metabolites generated from diverse tryptophan metabolic pathways show both beneficial and harmful effects.

Butyrate, one of the short-chain fatty acids (SCFAs), has shown vasodilation effects. We examined whether sodium butyrate administration in pregnancy and lactation can prevent hypertension induced by a maternal tryptophan-free diet in adult progeny, and explored protective mechanisms.

Decreased tryptophan metabolites indole-3-acetamide and indoleacetic acid observed in offspring born to dams that received the trytophan-free (TF) diet coincided with hypertension. This suggested that gut microbiota-derived tryptophan metabolites might be an offsetting mechanism, but not a cause of TF-induced hypertension. Considering that TF intervention reduced abundance of Romboutsia and Akkermansia, and many species are able to metabolize tryptophan, further studies linking abundance of bacterial species and concentrations of tryptophan metabolites are still required to identify main tryptophan metabolite producers.

Sodium butyrate treatment during pregnancy and lactation offset effects of maternal tryptophan-deficiency-induced offspring hypertension, mainly related to shaping gut microbiome, mediating SCFA receptor GPR41 and GPE109A, and restoring the renin–angiotensin system. A better understanding of mechanisms behind tryptophan metabolism implicated in programming of hypertension is critical for developing gut microbiota-targeted therapies to halt hypertension.”

https://www.sciencedirect.com/science/article/abs/pii/S0955286322001619 “Sodium butyrate modulates blood pressure and gut microbiota in maternal tryptophan-free diet-induced hypertension rat offspring” (not freely available) Thanks to Dr. You-Lin Tain for providing a copy.


A second study was on propionate effects:

“Early-life disturbance of gut microbiota has an impact on adult disease in later life. Propionate, one of predominant SCFAs, has been shown to have antihypertensive property.

We examined whether perinatal propionate supplementation can prevent offspring hypertension induced by maternal chronic kidney disease (CKD). CKD is closely linked to adverse maternal and fetal outcomes, and is reported to affect at least 3%-4% women of childbearing age.

Male offspring were divided into four groups: control, CKD, control+propionate (CP), and CKD+propionate (CKDP).

nutrients-14-03435-g001

Perinatal propionate supplementation:

  • Prevented offspring hypertension;
  • Shaped gut microbiota with increases in species richness and evenness;
  • Increased plasma propionate level; and
  • Upregulated renal GPR41 expression.

Results reveal the feasibility of manipulating gut microbiota by altering their metabolites with early-life use of propionate to prevent offspring hypertension in later life.”

https://www.mdpi.com/2072-6643/14/16/3435/htm “Perinatal Propionate Supplementation Protects Adult Male Offspring from Maternal Chronic Kidney Disease-Induced Hypertension”


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An inflammation clock

Here are six 2022 papers that either cited the second study of Variable aging measurements, or provided further evidence for its findings. Let’s start with a citing study:

“This study aimed to investigate expression patterns and prognostic values of the inflammatory aging clock (iAge) in glioblastoma (GBM), and its relations with stem cells. Similar to epigenetic clocks and transcriptomic clocks, iAge could track multifaceted aging phenotypes and have clinical significance in translation medicine.

iAge was positively correlated with chronological age, and highly associated with immune cells and inflammatory activities. iAge could serve as a prognostic biomarker for overall survival, and could precisely predict GBM stem cells stemness.

We identified the physiological importance and function of iAge in GBM, and provided novel insights into how iAge is a critical event for development of GBM.”

https://www.frontiersin.org/articles/10.3389/fgene.2022.925469/full “Inflammatory aging clock: A cancer clock to characterize the patients’ subtypes and predict the overall survival in glioblastoma”


Beginning with a human osteoporosis study, five papers investigated cytokine CXCL9, which the iAge study found to be “clearly actionable as shown by our experiments in CXCL9 where we can reverse aging phenotypes.”

“We assessed whether levels of CXCL9 and CXCL10 were elevated in human serum samples of older adults who had incident hip fractures. Our findings revealed higher serum levels of CXCL9 in pre-fracture blood samples of men with subsequent hip fractures, compared with their non-fracture controls. There was no such difference in CXCL9 serum levels between cases and controls in women.

Serum CXCL9 improved the prediction of osteoporotic hip fracture in men. The association between CXCL10 and hip fracture risk was not statistically significant in either sex.

While our epidemiologic findings are supported by experimental data providing the mechanistic pathway for CXCL9 in regulating osteoclast recruitment, further studies are needed to confirm validity of our findings and determine their generalizability to other study populations. Underlying biological mechanisms that limit our findings to men but not women require further investigation.”

https://asbmr.onlinelibrary.wiley.com/doi/10.1002/jbmr.4646 “CXCL9 Predicts the Risk of Osteoporotic Hip Fracture in a Prospective Cohort of Chinese Men—A Matched Case–Control Study”


Two immune-mediated skin diseases, with a vitiligo review:

“Current findings emphasize the critical role of immune cells and their mediators in the immunopathogenesis of vitiligo. IFN-γ [interferon gamma] is the primary cytokine mediator that activates the JAK/STAT pathway, causing keratinocytes to produce the key chemokines CXCL9 and CXCL10.

Interactions between immune and non-immune cells finally result in apoptosis of melanocytes. Additional investigations of these pathways may provide an opportunity for finding possible therapeutic targets, as there are currently no targeted biological drugs available for treatment of vitiligo.”

https://www.mdpi.com/2227-9059/10/7/1639/htm “Current Concepts of Vitiligo Immunopathogenesis”

and a study of psoriasis:

“CXCL9 is an important chemokine involved in T cell recruitment, and is up-regulated in plasma of patients with psoriasis. Increased CXCL9 expression can aggravate the progression of psoriasis.

cxcl9 expression

IL-1β and CXCL9 were up-regulated and CLDN8 was down-regulated in psoriasis with statistically significant differences. Identification of potential key molecular markers and signaling pathways provides potential research directions for further understanding molecular mechanisms of psoriasis.”

https://www.wjgnet.com/2307-8960/full/v10/i18/5965.htm “Identification of potential key molecules and signaling pathways for psoriasis based on weighted gene co-expression network analysis”


Two lung-related studies, first, an editorial for a human lung transplant study that isn’t freely available:

“CXCL9 and CXCL10 are chemokines that bind to the shared receptor CXCR3, potentiating T cells, mononuclear cells, and natural killer (NK) cells. Previous studies demonstrated that presence of these chemokines in bronchoalveolar lavage samples preceded development of chronic lung allograft dysfunction (CLAD).

Acute rejection and acute lung injury are known risk factors to the development of CLAD, yet this study found that increased risk was dependent on the presence of CXCL9/CXCL10 plasma elevation. Early identification of patients at risk, possibly during the active inflammatory phase, rather than once abnormal wound healing pathways dominate resulting in irreversible injury, provides an attractive opportunity for intervention.”

https://onlinelibrary.wiley.com/doi/10.1111/ajt.17135 “CXCL9 and CXCL10 plasma levels: Potential keys to unlocking CLAD risk”

and a study of smoking effects:

“We collected blood samples from 78 healthy male volunteers aged 18–60, including non-smokers (n = 30), current smokers (n = 30), and ex-smokers (n = 18). Expression levels of CXCL9/MIG [monokine induced by IFN-γ] and sIL-6R significantly increased after smoking, and continued to increase after quitting smoking.

cxcl9 smoking

Changes in related cytokines after smoking cessation are mainly restorative, while some cytokines further strengthen the trend of smoking-related changes.”

https://www.mdpi.com/1420-3049/27/12/3715/htm “Effects of Smoking on Inflammatory-Related Cytokine Levels in Human Serum”


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Natural sulforaphane effects

This 2022 rodent cell study used the natural form of sulforaphane to replicate experiments performed with mixtures of its natural and unnatural forms:

“Natural sulforaphane (SFN) exists as a single enantiomer with a RS absolute configuration. Most studies focusing on its biological activities, in particular its anti-inflammatory and antioxidant activities, have been conducted using its racemic (rac) form. rac-SFN has shown these effects in several in vitro and in vivo models.

(R)-sulforaphane

These findings demonstrate that (R)-SFN was able to:

  • Modulate inflammatory response and oxidative stress induced by LPS stimulation in murine peritoneal macrophages;
  • Reduce pro-inflammatory enzyme expression (iNOS, COX-2 and mPGES-1) and cytokine production (IL-1β, IL-6, IL-17, IL-18 and TNF-α);
  • Inhibit MAPK, JAK2/STAT-3, and canonical and non-canonical inflammasome signaling pathways;
  • Reduce NO and ROS levels and up-regulate the Nrf-2/HO-1 axis; and
  • Modulate epigenetic changes through histone methylation (H3K9me3) and deacetylation (H3K18ac).

(R)-SFN could be a new epinutraceutical compound useful for management of several immunoinflammatory diseases.”

https://www.mdpi.com/1424-8247/15/8/966/htm “Immunomodulatory Effects of (R)-Sulforaphane on LPS-Activated Murine Immune Cells: Molecular Signaling Pathways and Epigenetic Changes in Histone Markers”


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Epigenetic effects of plasma concentrate

“We use data from a safety study (n = 18, mean age 74) to investigate whether human umbilical cord plasma concentrate (hereinafter Plasma Concentrate) injected weekly (1 ml intramuscular) into elderly human subjects over a 10-week period affects different biomarkers, including epigenetic age measures, standard clinical biomarkers of organ dysfunction, mitochondrial DNA copy number (mtDNA-CN), and leukocyte telomere length.

More than 20 clinical biomarkers were significantly and beneficially altered. Telomere length and mtDNA-CN were not significantly affected by treatment.

An increase in entropy means that the methylome becomes noisier. We found that entropy was significantly decreased after treatment. Decreased entropy may implicate rejuvenation of the epigenetic landscape after plasma concentrate treatments.

changes in methylation entropy

Treatment reduced DNA methylation-based GrimAge by an average of 0.82 years, suggesting a reduction in morbidity and mortality risk. By contrast, no significant results could be observed for epigenetic clocks that estimate chronological age.

Our study lends credence to the notion that there are youth-promoting factors in the secretome of umbilical cord plasma. This conclusion has also been reached by other researchers that have provided treatment with stem cells, which do not work by plasma dilution but primarily by providing humoral factors and changing the microenvironment of cells and tissues. While there may be youth-promoting microvesicles or humoral factors that are at work, we do not want to rule out the possibility that it is ‘young and undamaged’ albumin that leads to the improvements noted, especially in light of recent evidence for such a mechanism.

This first human epigenetic clock study of plasma concentrate treatments revealed age-reversal effects according to a well-established DNA methylation-based estimator of morbidity and mortality risk. Future placebo-controlled replication studies are warranted with a larger number of participants over a longer study period, which our laboratory has undertaken to pursue.”

https://onlinelibrary.wiley.com/doi/10.1111/acel.13696 “Umbilical cord plasma concentrate has beneficial effects on DNA methylation GrimAge and human clinical biomarkers”


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Improving dietary fiber research

Two 2022 papers on aspects of dietary fiber research, starting with a review:

“Considerable attention has been given to unraveling the interaction between fiber type and gut microbiota utilization, focusing mainly on single, purified fibers. Studying these fibers in isolation might give us insights into specific fiber effects, but neglects how dietary fibers are consumed daily and impact our digestive tract: as intrinsic structures that include the cell matrix and content of plant tissues.

Food processing per se is not health-detrimental. Certain foods are barely digestible without any processing, and for specific populations, e.g. those suffering from malnutrition or diseases, food processing is crucial. However, increased digestibility has resulted in negative health outcomes related to obesity and welfare diseases.

whole grain

Intrinsic structural features of plant cells likely slow down fiber fermentation, inducing a lag phase, but do not necessarily reduce the absolute amount of short-chain fatty acids (SCFA) produced. Consequently, there is a gradual release of SFCA, which means that SCFA production is not restricted to the proximal colon but spread throughout the whole colon, including its distal parts, benefiting local, mucosal health.

This translates into beneficial, systemic, peripheral effects as distal SCFA infusion in vivo has shown to induce more pronounced effects on biomarkers than proximal. Delayed fermentation of intrinsic fibers presents a highly relevant feature that isolated, single fibers do not have.

Instead of further processing our already extensively processed foods to create new products, we should minimize this processing, and exploit health benefits associated with the original cell matrix of plant tissues.”

https://www.frontiersin.org/articles/10.3389/fimmu.2022.954845/full “Intrinsic dietary fibers and the gut microbiome: Rediscovering the benefits of the plant cell matrix for human health”


Reference 115 was a human study by the same researchers:

“We investigated the impact of dried chicory root in a randomised, placebo-controlled trial with 55 subjects at risk for type 2 diabetes. Evidence for a trophic chain including Bifidobacterium and Anaerostipes spp. was recapitulated by in vitro incubations that resulted in high levels of butyrate and propionate production from the treatment product.

butyrate and propionate microbial network

We observed a simultaneous increase in faecal and circulating SCFA levels, and a marked improvement in dynamic markers of glucose control. In subjects with a low relative abundance of Blautia spp. – a genus that previously has been associated with T2D – static glycaemic markers also decreased pronouncedly.

Our results demonstrate a strong modulatory potential on gut health and microbial metabolism by native inulin and cell wall fibres pectin, cellulose and hemicellulose in the intrinsic form of dried chicory roots.”

https://www.cambridge.org/core/journals/gut-microbiome/article/dried-chicory-root-improves-bowel-function-benefits-intestinal-microbial-trophic-chains-and-increases-faecal-and-circulating-short-chain-fatty-acids-in-subjects-at-risk-for-type-2-diabetes/6209AEAFBDDB181197F22AE24388186B# “Dried chicory root improves bowel function, benefits intestinal microbial trophic chains and increases faecal and circulating short chain fatty acids in subjects at risk for type 2 diabetes”


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Garlic vs. broccoli

This 2022 human study compared effects of two supplements:

“We test the hypothesis that consuming glucoraphanin (from broccoli) or alliin (from garlic) results in the accumulation of sulforaphane and alliin and their associated metabolites in the human prostate gland in a randomised, double-blinded, 2 × 2-factorial, dietary supplement, four-week intervention study.

The predominant sulphur-containing metabolite in garlic is alliin, which is odourless and non-volatile. When the plant tissue is damaged, alliinase enzymes rapidly convert alliin to allysulfenates that condense to form allicin and other thiosulfinates, predominantly γ-glutamyl S-allyl-L cysteine (γ-SAC) and S-allyl-L cysteine (SAC).

The BroccoMax/GRN supplements (530 mg) contained 97.7 ± 6.70 µmol glucoraphanin. The Kwai/alliin supplements (715 mg) contained four garlic-derived metabolites: alliin (35.2 ± 0.52 µmol), γ-SAC (19.3 ± 1.91 µmol), SAC (1.8 ± 0.16 µmol), and allicin (21.4 ± 2.10 µmol).

Mean excretion of sulforaphane and its metabolites as a percentage of ingested glucoraphanin [aka bioavailability] was 56.21% (range 21–91%, SD ± 18.66).

sulforaphane bioavailability

Alliin was detected within the prostate of every participant. Estimation of dietary intake of alliaceous vegetables is challenging due to their widespread presence in processed foods, and it is likely that intake is often underestimated.

We provide evidence that sulforaphane can be detected in human prostate tissue following regular consumption of glucoraphanin supplements. In contrast, alliin and associated metabolites were not more abundant in prostates of men receiving the alliin garlic-derived supplement. It is conceivable that alliin does accumulate in human prostate tissue, but its turnover is much slower than that of sulforaphane so that a longer allium-free diet is required prior to an intervention to assess its accumulation.

Accumulation of sulforaphane and presence of alliin in prostate tissue, as demonstrated in this study, may result in local effects on healthy and cancerous cells through a variety of mechanisms. This may explain the reduced risk of prostate cancer incidence and progression following consumption of cruciferous and alliaceous vegetables.”

https://www.mdpi.com/2072-6643/14/16/3263/htm “Accumulation of Sulforaphane and Alliin in Human Prostate Tissue”


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Another use of a routine blood test

A 2022 human study investigated three measurements of the CBC with Differential/Platelet test:

“This study explored the relationship between the systemic immune-inflammation index (SII) and stroke prognosis. SII was defined as neutrophils × platelets/lymphocytes. Neutrophils and lymphocytes are involved in the inflammatory and immune response, whereas platelets have a primary role in the thrombo-inflammation of stroke.

Patients were divided into four groups according to SII values: quartile (Q)1 <366; Q2 366–533; Q3 534–799; and Q4 ≥800. As the SII quartile increased, patients with acute ischemic stroke were more likely to have poor functional outcomes during follow-up.

Although age, smoking status, and alcohol consumption are risk factors, and female estrogen is a protective factor for stroke, we did not identify subgroups specifically being affected by these factors. Even if we adjusted our model for factors identified by previous studies as immune-inflammation markers that might affect prognosis, such as the high-sensitivity C-reactive protein level, our results still suggested that SII is closely related to short- and long-term prognosis of patients with acute ischemic stroke.

As a new type of immune-inflammation index, the SII integrates neutrophils, platelets, and lymphocytes, and can reflect the balance of the systemic immune response and inflammatory response.”

https://www.aging-us.com/article/204228/text “Correlation of the systemic immune-inflammation index with short- and long-term prognosis after acute ischemic stroke”


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Epigenetic clocks so far in 2022

2022’s busiest researcher took time out this month to update progress on epigenetic clocks. If I curated every study he’s contributed to, it would require at least three blog posts a week. I’ll link to a few he’s posted in August 2022 that are more appreciated in the researcher community.

“In my lab, we are looking for clocks that apply to multiple species at the same time, for example, universal pan-mammalian clocks. It’s all about enhancing translation.

If you have an intervention that rejuvenates a mouse, a rat, a dog, and a cat according to the same clock, then chances are high that it will also work in humans. Naked Mole-Rat Hyaluronan Synthase 2 Promotes Longevity and Enhances Healthspan in Mice

Several groups, including mine, are working on single cell methylation clocks. Researchers are building clocks that respond to lifestyle interventions, such as exercise.

Moving away from methylation, it would be nice to build similar clocks for other ‘omics’ data. Many researchers build clocks on the basis of other omics data, such as for chromatin, proteomics, and gene expression.

There are different platforms, but they all attempt to measure the same thing: biological age. LINE-1 RNA causes heterochromatin erosion and is a target for amelioration of senescent phenotypes in progeroid syndromes

Epigenetic clocks are ‘life course clocks.’ I don’t know any other biomarkers of aging that applies to fetal tissues as well, because most other biomarkers measure organ dysfunction. Epigenetic profiling and incidence of disrupted development point to gastrulation as aging ground zero in Xenopus laevis

There’s this company called Intervene Immune, founded by Greg Fahy, and they are using GrimAge and other epigenetic clocks in clinical trials. They are doing a Phase II clinical trial. By the way, I’m one of the participants.

I could name several other groups who are using epigenetic clocks in clinical trials. It would be interesting if more people would measure epigenetic age in clinical trials in humans, at least as a secondary outcome, because there’s always an opportunity to make a discovery.

If you compare GrimAge to other biomarkers, such as cholesterol or glucose levels, you will see similar noise levels there. Epigenetic clocks are remarkably robust compared to what else is used in the clinic. I would say that the issue with technical noise in epigenetic clocks has been solved.

I’m really glad that different companies and researchers pursue different avenues, since it diversifies our risk. If one of these approaches works, it will change the world.”

https://www.lifespan.io/news/steve-horvath-on-the-present-and-future-of-epigenetic-clocks/ “Steve Horvath on the Present and Future of Epigenetic Clocks”


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Glucoraphanin is not sulforaphane

A poorly-conceived and intentionally-misrepresented human 2022 broccoli product study:

“We investigated whether a sulforaphane (SFN) [actually, sulforaphane precursor glucoraphanin] intake intervention improved cognitive performance and mood states in healthy older adults in a 12-week, double-blinded, randomized controlled trial.

The SFN group showed improvement in processing speed and a decrease in negative mood compared to the placebo group. However, there were no significant results in other biomarkers of oxidant stress, inflammation, or neural plasticity.

These results indicate that nutrition interventions using SFN can have positive effects on cognitive functioning and mood in healthy older adults.”

https://www.frontiersin.org/articles/10.3389/fnagi.2022.929628/full “Effects of sulforaphane intake on processing speed and negative moods in healthy older adults: Evidence from a randomized controlled trial”


Contrary to this study’s title, actual sulforaphane intake was not measured. The glucoraphanin product used in this study was the same item and daily dose as Eat broccoli sprouts for your workouts, which investigated effects with 19-to-23-year-old men. The treatment was taken all at once at an unspecified time of day rather than three times a day with young subjects.

These researchers knew from the 2012 study cited for dose that:

“Individual conversions of glucosinolates [like glucoraphanin] to isothiocyanates [like sulforaphane] varied enormously, from about 1% to more than 40% of dose. In contrast, administration of isothiocyanates (largely sulforaphane)-containing broccoli sprout extracts, resulted in uniformly high (70-90%) conversions to urinary dithiocarbamates.”

Young or old, a daily 30 mg glucoraphanin intake isn’t sufficient to fully activate human Nrf2 signaling pathways. A daily 17 mg sulforaphane intake could accomplish that.


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Don’t bother eating broccoli sprouts if you’re old?

I try to not curate research that wastes resources. Couldn’t help but present this 2022 rodent study:

“We aimed to evaluate if sulforaphane (SFN) long-term treatment was able to prevent age-associated cognitive decline in adult (15-month-old) and old (21-month-old) female and male rats.

Our results showed that SFN restored redox homeostasis in brain cortex and hippocampus of adult rats, preventing cognitive decline in both sexes. However, redox responses were not the same in males and females.

Old rats were not able to recover their redox state as adults did, but they had a mild improvement. These results suggest that SFN mainly prevents rather than reverts neural damage; though, there might also be a range of opportunities to use hormetins like SFN, to improve redox modulation in old animals.”

https://link.springer.com/article/10.1007/s10522-022-09984-9 “Long-term sulforaphane-treatment restores redox homeostasis and prevents cognitive decline in middleaged female and male rats, but cannot revert previous damage in old animals” (not freely available)


These researchers cited Sulforaphane in the Goldilocks zone for hormetic effects of sulforaphane, so I asked:

“Did you develop any preliminary dose/response data for stating ‘there might also be a range of opportunities to use hormetins like SFN to improve redox modulation in old animals’?”

They cited Broccoli sprouts activate the AMPK pathway for long-term effects of a small sulforaphane dose, so I asked:

“Also, the three studies cited for ‘0.5 mg/Kg, i.e. 2.82 μmol/Kg BW for 3 months’ were all mouse studies. Since this was a rat study, wouldn’t there be increased dose and duration equivalencies?”

I’ll update this blog post in the event either of my questions to these researchers are answered.

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Sulforaphane nose drops

This 2022 rodent study compared capabilities of intranasal nanoparticle sulforaphane and free sulforaphane to mitigate brain damage caused by a common cancer treatment:

“Non-invasive intranasal (IN) trafficking of therapeutic agents with nanocarriers can enhance efficacy of drug delivery, biodistribution, bioavailability, and absorption against enzymatic degradation and extracellular transportation. Direct IN trafficking of nanocarriers is expected to reduce drug wastage, administration frequency, and undesirable adverse effects.

The nasal route for brain-targeted delivery of sulforaphane (SF) loaded within iron oxide nanoparticles (Fe3O4-NPs) was based on improving physicochemical stability of SF, and to enhance its bioavailability by avoiding oral route drawbacks like extensive first-pass metabolism and intestinal drug degradation.

Cisplatin (CIS) significantly induced a significant increase in acetylcholinesterase activities and lipid peroxides, and a significant decrement in glutathione and nitric oxide contents. We aimed to explore the nanotherapeutic potential of intranasally delivered SF loaded within Fe3O4-NPs (N.SF) against CIS-induced neurotoxicity through different biochemical, behavioral, and histological investigations.

hippocampus damage

Treatment with N.SF was more capable of mitigating both CIS-induced striatal and cortical injuries. IN treatment with either SF or N.SF showed equal alleviative potential regarding CIS-induced hippocampal or cerebellar injury.

These encouraging results demonstrated the potential use of iron-oxide NPs as neurotherapeutic agents, and confirmed the possibility of developing a novel promising and non-invasive intranasal delivery system for treatment of CIS-induced neurotoxicity.”

https://link.springer.com/article/10.1007/s12640-022-00555-x “Neuroprotective Potential of Intranasally Delivered Sulforaphane-Loaded Iron Oxide Nanoparticles Against Cisplatin-Induced Neurotoxicity”


I found this study from it citing a paper in Do broccoli sprouts treat migraines?

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Non-patentable boron benefits

To follow up Is boron important to health? I’ll highlight a 2022 review of boron intake:

“Boron is essential for activity of several metabolic enzymes, hormones, and micronutrients. It is important for growth and maintenance of bone, reduction in inflammatory biomarkers, and increasing levels of antioxidant enzymes.

The average person’s daily diet contains 1.5 to 3 milligrams of boron. Boron intakes of 1–3 mg/day have been shown to improve bone and brain health in adults when compared to intakes of 0.25–0.50 mg/day.

One week of 10 mg/d boron supplementation resulted in a 20% reduction in inflammatory biomarkers TNF-α, as well as significant reductions (nearly 50%) in plasma concentrations of hs-CRP and IL-6. Calcium fructoborate, a naturally occurring, plant-based boron-carbohydrate complex, had beneficial effects on osteoarthritis (OA) symptoms. A double-blind study in middle-aged patients with primary OA found that all groups except the placebo group saw a reduction in inflammatory biomarkers after 15 days of food supplementation with calcium fructoborate.

Dietary boron intake significantly improves brain function and cognitive functioning in humans. Electroencephalograms showed that boron pharmacological intervention after boron deficiency improved functioning in older men and women, such as less drowsiness and mental alertness, better psychomotor skills (for example, motor speed and dexterity), and better cognitive processing (e.g., attention and short-term memory). Boron compounds can help with both impaired recognition and spatial memory problems.

We discussed the role of boron-based diet in memory, boron and microbiome relation, boron as anti-inflammatory agents, and boron in neurodegenerative diseases. Boron reagents will play a significant role to improve dysbiosis.”

https://www.mdpi.com/1420-3049/27/11/3402/htm “The Role of Microbiome in Brain Development and Neurodegenerative Diseases”


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