How to measure biological age?

As mentioned in Week 127, I had biological age measured earlier this month, and received five reports two days ago on Sunday. Part of the company’s process is to follow up their reports (intrinsic aging, immune aging, pace of aging, telomere length, weight loss) with a consulting session to review and interpret, which lasted an hour yesterday.

Part of our conversation revolved around comparing my measurements with other customers. These people are a different population than people usually sampled for aging and other biomarkers, because people who pay to get their biological age measured probably actionably want to improve it.

We’ll see which items I asked the consultant to pass on to the company produce responses, and which interfere with their business or they’re too busy to get back to me. I offered more than a half-dozen specifics, but held back on items I didn’t think the consultant would adequately communicate.

I didn’t argue with the consultant’s recommendations for quercetin supplementation (at 4% bioavailability?) as part of a treatment for senescence (not measured in any of the reports?). I didn’t offer to follow-up with studies demonstrating yeast cell wall β-glucan (new to the consultant) effects on immune report findings here in my 19th year of taking it every day.

I did argue with their recommendation to take DHEA-S. I changed my mind about taking it a year and a half ago ago, but left blog posts up such as Take responsibility for your one precious life – DHEA for evidence that I’m learning.

Epigenetic clocks per The epigenetic clock theory of aging generally view biological aging as “an unintended consequence of both developmental programmes and maintenance programmes, the molecular footprints of which give rise to DNAm [DNA methylation] age estimators.”

So what would be appropriate anti-aging actions for customers to take? Should customers try to emulate youthful biological markers, and supplement DHEA-S to impact serum levels of insulin-like growth factor 1?

I don’t think so. Our bodies never evolved feedback mechanisms to determine “Time to stop the growth programs, you’ve survived to reproduction age.” Older people achieving teenagers’ DHEA-S levels and activating IGF-1 pathways, pretty much guarantees further biological aging as “an unintended consequence of both developmental programmes and maintenance programmes.”


It’s too early to recommend these biological aging measurements. We’ll see where it goes.

One good thing is the company wants their customers to tell them everything about what they’re doing. I exercise at least a half hour every day, eat Avena nuda oats for breakfast and AGE-less chicken vegetable soup for dinner, and take the following:

Before breakfast
– 3-day-old microwaved broccoli / red cabbage / mustard sprouts started from 10.7 grams of seeds, with nothing else an hour before or after
– Yeast cell wall β-glucan (Glucan 300), 1500 mg, with nothing else an hour before or after
– Calcium alpha-ketoglutarate 1 g

Breakfast, lunch, and dinner
– Hyaluronic acid, Nature’s Lab, 1 serving total
– Boron, Swanson Triple Boron Complex, 9 mg total

Breakfast and dinner
Acetyl-L-carnitine, 1 g total
– Balance oil, which blends linoleic acid 1400 mg with linolenic acid 350 mg, 2 times
– Betaine anhydrous, 3 g total
– Glucosamine hydroxychloride 1.5 g total, with chondroitin sulfate 1.2 g total
Taurine, 2 g total
– 3-day-old Avena sativa oat sprouts started from 20 g seeds, 2 times

Breakfast only
– Minerals and vitamins, RDA mainly, Kirkland Signature Daily Multi
– D3 25 mcg

Lunch only
– Vitamin K2 MK-7 600 mcg

Dinner only
– D3 50 mcg
– Zinc monomethionine 30 mg with 0.3 mg copper
– Lutein 25 mg with 5 mg zeaxanthin


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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 – How to measure biological age?

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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