Category: aging

Choe YM, Suh GH, Lee BC, Choi IG, Lee JH, Kim HS, Kim JW. Association Between Copper and Global Cognition and the Moderating Effect of Iron. Front Aging Neurosci. 2022 Mar 29;14:811117. PMC free article

This is a really nice study out of South Korea that suggests that blood serum copper and iron in the low end of the reference range may be associated with cognitive decline as measured by the the Mini-Mental State Examination (MMSE) We need both copper and iron in our carrier protein ceruloplasmin.

On this site we have mad much ado about the active role of copper in Complex IV of the mitochondria electron transport chain. If one checks the Wikipedia site on Complex IV function, it becomes clear that iron also plays a very important role in electron transport. Let us just keep in mind while we review the South Korean study that Cu and Fe are partners in traversing our circulation and generation of ATP in our mitochondria.

This study was part of a larger longitudinal study that follows patients over time, in this 99 non-demented adults between 65 and 90 years of age: 42 cognitively normal (CN) adults and 57 adults with mild cognitive impairment (MCI) were enrolled in the General Lifestyle Alzheimer’s Disease study with 13 dropouts. The Clinical Normal group consisted of participants with a Clinical Dementia Rating score of 0 and no diagnosis of MCI or dementia. All participants with Mild Cognitive Impairment met the current consensus criteria for amnestic MCI. Patients with mental illnesses and other physical issues associated with test taking were excluded.

Clinical Assessments

The MMSEwas used to measure global cognitive function. Vascular risk factors, income, smoking status, and so on were also taken into consideration.

Measuring Serum Copper Levels and Other Blood Biomarkers

Blood was collected after overnight fasting in the morning (8–9 a.m.). The normal range for the serum copper level is 75–145 μg/dL according to the Mayo Clinic Laboratories. This reference category was used to subdivide the participants into three groups, or tertiles.

• low (< 87 μg/dL)
• medium (87–98 μg/dL)
• high (> 98 μg/dL)

Blood iron was divided into “low” and “high” according to the reference range.

Table 1 asked if smoking status, education, income, alcohol consumption, gender,APO4E and so on predicted being in the low, medium, or high copper group. They did not by themselves. The APOE gene codes for apolipoprotein. The APO4E variant, one of three, of apolipoprotein puts one at risk for Alzheimer’s Disease. “Version 4 is less effective at removing plaques from the brain. Individuals who have version 4 have a significantly increased risk for developing Alzheimer’s,” according to Alzheimer’s Organization. The authors came up with models (equations) to incorporate factors that might affect the MMSE score and serum copper. These other factors are confounding variables. Let us say that people who drink a lot have a higher risk of lung cancer. Does alcohol cause lung cancer? Maybe not. People who drink a lot may hang out in bars where there is a lot of tobacco smoke. Perhaps those who drink a lot are also addicted to tobacco as well. We need not concern ourselves with the equations the South Korean group used to sort out the other life style issues that influence MMSE score and serum copper and iron.

We’ve covered a confounder. Multiple linear regression means taking multiple predictors into the outcome variable MMSE. The lower the P value, the higher the probability that the observed difference is not due to random chance. The cut off of significance is generally p<0.05, or 95% certain that random chance was not at play. The R² value, Pearman correlation coefficient, tells us how tight the dependent outcome variable is with the independent variable. A value of 1.0 tells us there is a one to one tightness of one with the other. A value of 0 means no relation whatsoever.

• Overall, the higher the adjusted serum copper, the higher the MMSE…. serum iron notwithstanding. We are very sure that this observation is not due to random chance. The tightness of the relationship is sort of borderline.
• When we go to the low iron group, we see the same increase in MMSE score as a function of serum copper. The R² value is closer to 1.0 and the p value indicates that this is not due to random chance.
• In the high serum iron group, serum copper is less of a predictor of MMSE score.

Then the publication moved on to bar graphs that might be easier to understand.

When a person is in the low iron group, going from low to high copper makes a significant difference in the MMSE score. If the elder already has high iron, being low or high copper does not make that much difference in the MMSE score.

If an elder is in the low reference range for copper and iron, should he/she supplement? This would be a health care provider question. The Choe study from South Korea suggests every little bit helps.

If we were to conduct an informal version of this study, we may not include many confounding variables if we are starting with seniors with low reference range copper and/or iron. If we are just looking a cognitive scores while in the low reference range for copper and the after scores we need not worry as much about confounding variables. ApoE4 status will not change. Smoking and alcohol use will probably not change. We could add an additional outcome variable like “chasing grand kids.” We’d then have to measure the copper and/or iron status after a given period of supplementation.

This post started out exploring an NAD⁺ boosting niacin clinical trial. The thought that if some other group had shown that niacin increases NAD⁺ we don’t need to prove it. Key enzymes in the NAD salvage pathway play a role in a progressive external opthalmoplegia (PEO) as shown by a niacin clinical trial. Then a second request came in to explore telomeres and the role NAD⁺ has in controlling telomere length.

1. What is PEO and the niacin link?
   1. from ncbi conditions
   2. from the Pirinen 2020 publication
2. NAD+ and telomeres
   1. What are telomeres and telomerase?
   2. A mammalian telomere, NAD+ comes in here
   3. Back to yeast telomeres and NAD+ salvage
3. yeast telomere references

What is PEO and the niacin link?

Pirinen E, Auranen M, Khan NA, Brilhante V, Urho N, Pessia A, Hakkarainen A, Kuula J, Heinonen U, Schmidt MS, Haimilahti K, Piirilä P, Lundbom N, Taskinen MR, Brenner C, Velagapudi V, Pietiläinen KH, Suomalainen A. Niacin Cures Systemic NAD⁺ Deficiency and Improves Muscle Performance in Adult-Onset Mitochondrial Myopathy. Cell Metab. 2020 Jun 2;31(6):1078-1090.e5. PMC free article

The following are some definitions to consider before reviewing a clinical trial to improve this condition with niacin supplementation.

from ncbi conditions

“Progressive external ophthalmoplegia is a condition characterized by weakness of the eye muscles. The condition typically appears in adults between ages 18 and 40 and slowly worsens over time. The first sign of progressive external ophthalmoplegia is typically drooping eyelids (ptosis), which can affect one or both eyelids. As ptosis worsens, affected individuals may use the forehead muscles to try to lift the eyelids, or they may lift up their chin in order to see. Another characteristic feature of progressive external ophthalmoplegia is weakness or paralysis of the muscles that move the eye (ophthalmoplegia). Affected individuals have to turn their head to see in different directions, especially as the ophthalmoplegia worsens. People with progressive external ophthalmoplegia may also have general weakness of the muscles used for movement (myopathy), particularly those in the neck, arms, or legs. The weakness may be especially noticeable during exercise (exercise intolerance). Muscle weakness may also cause difficulty swallowing (dysphagia). When the muscle cells of affected individuals are stained and viewed under a microscope, these cells usually appear abnormal. These abnormal muscle cells contain an excess of cell structures called mitochondria and are known as ragged-red fibers. ”

from the Pirinen 2020 publication

Progressive External Ophthalmoplegia is generalized muscle weakness, and susceptibility to fatigue is often caused by single or multiple mitochondrial DNA (mtDNA) deletions.
A mouse model for some progressive mitochondrial myopathy manifests as NAD⁺ depletion. Intracellular NAD⁺ concentrations can be increased by various approaches, such as decreasing NAD⁺ consumption and NAD⁺ precursor supplementation: nicotinic acid (niacin), nicotinamide (NAM), and nicotinamide riboside (NR). The simplest approach would be to supplement with niacin and also examine lipid metabolic parameters that have been shown to occur with niacin supplementation and that might relate to NAD⁺

The design

This study came out of the University of Helsinki, Finland. These researchers at the University of Helsinki report that mitochondrial muscle disease leads to low NAD⁺ levels in both blood and muscle. They treatment treated niacin, a vitamin B3 form and an NAD⁺ precursor, improves NAD⁺ levels, disease signs, and muscle metabolism in patients, also improving muscle strength and performance. These results indicate that NAD⁺ depletion occurs in human diseases, and its repletion is a potential therapy for mitochondrial myopathies. Patients were given escalating doses of administered an increasing doses of the NAD⁺-booster niacin form (to 750-1,000 mg/day.

Arm	Intervention/treatment
Experimental: Niacin in controls The arm includes healthy controls supplemented with niacin.	Dietary Supplement: Niacin The dose for a slow-released form of niacin will be 750-1000 mg/day. The daily niacin dose, 250 mg/day, is gradually escalated by 250 mg/month so that the full dose is reached after 3 months. The intervention time with the full niacin dose is 1 and 7 months for controls and patients, respectively, and subsequently total intervention time 4 and 10 months, respectively. At the end of the study, the daily dose will be decreased by 250 mg/month rate. Other Name: Nicotinic acid
Experimental: Niacin in mitochondrial myopathy patients The arm includes mitochondrial myopathy patients supplemented with niacin.	Dietary Supplement: Niacin The dose for a slow-released form of niacin will be 750-1000 mg/day. The daily niacin dose, 250 mg/day, is gradually escalated by 250 mg/month so that the full dose is reached after 3 months. The intervention time with the full niacin dose is 1 and 7 months for controls and patients, respectively, and subsequently total intervention time 4 and 10 months, respectively. At the end of the study, the daily dose will be decreased by 250 mg/month rate. Other Name: Nicotinic acid

Fig. 1 NAD⁺ chemical family changes

As a point of reference, the CopperOne clinical trial participants took 12.12mg cuprous niacin per day for 28 weeks. About 9 mg of this was niacin. The PEO clinical trial used much higher amounts of niacin.

Note that the PEO patients have lower muscle and whole blood NAD⁺ levels than the normal controls. Niacin supplementation increases the NAD⁺ in whole blood but not the muscle in the same controls. This panel is from the supplemental data section. It shows the enzymatic pathways between niacin (NA), NAD⁺, and other related small molecules. Are PEO patients the way they are because of enzyme deficiencies in these pathways?

Note that sirtuins are some of the NAD⁺ consuming enzymes. Nicotinamide riboside is not that different between PEO patients and controls. NADP and NMN are a little lower in PEO patients than the controls even after supplementation with large amounts of niacin. It is the NAM and ADPR that are much higher in the PEO patients even after supplementation.

NNMT catalyzes the reaction

S-adenosyl-L-methionine + nicotinamide ⇌ S-adenosyl-L-homocysteine + 1-methylnicotinamide.

These data are from supplemental figures of the publication. The authors were looking at gene transcripts for enzymes for proteins in NAD pathways. Note that NNMT enzyme activity is much higher in the PEO patients than the healthy controls. Not surprisingly, muscle NAM is much lower at baseline and at 4 months. Ten months of niacin supplementation seems to rectify the disparity.

Fig. 2 Lipid metabolism parameters

We became interested in the relationship between niacin and copper in fat metabolism and milk production when asked if our supplement would boost milk production in cows. See the fatty liver in cows post. While body weight was not affected, the authors saw variable decreases in fat stores. The reduction in liver fat in the PEO patients is probably most notable.

While increased muscle mass in normal and PEO patients may not be the same thing as milk production, we are extremely excited by this finding in humans. Adiponectin is a peptide hormone secreted by adipocytes. It controls fatty acid and glucose metabolism in many tissues. The energy expenditure is in units of percent of control.

Fig 3. Physical Function

Data in panels 3B- F were collected on a Good strength Metitur adjustable dynamometer chair. This is a Finnish brand. This chair measured static strength of back muscles, and dynamic strength of abdominal and shoulder muscles. FC stands for fraction of control.

Note the high variability in the PEO patients’ response to niacin. In many cases the supplemented PEO patients performed better than the controls!

The six minute walking test speaks more to the mitochondrial function that targets our customer base. As a reminder, a cartoon of pyruvate entering the NADH generating TCA cycle is shown. NADH supplies the ATP generating electron transport chain (ETC) of the mitochondria with electrons and H⁺. How much does the availability of NAD⁺/NADH + H⁺ dictate whether pyruvate goes to acetylCoA or lactate?

Note the drastic improvement in the six minute walk test in the PEO patients and the variable response in the control patients.

Fig. 4 the mitochondria

Cytochrome c oxidase (COX), respiratorycomplex IV of the electron transport chain, has some subunits encoded by mitochondrial DNA. Succinate dehydrogenase (SDH) complex II of the ETC is entirely encoded by nuclear genes. In PEO patients niacin decreased the fibers that were negative for COX yet positive for SDH. Fibers that were negative for complex I (CI) were also decreased. Mitochondrial total COX activity and mass in muscle fibers increased. Mitochondrial mass was also increased in both controls and PEO patients.

FC is fraction of control. In panels C and D we see that four months on niacin increases Cox activity and mitochondrial mass. We have to ask ourselves if increases like these can be seen with less “medicinal” levels of niacin and Cu(I).

No further comments will be presented on these SEM images on this post.

Fig 5. Metabolites

This post is going to skip the Principle Component Analysis of panel 5A. In short the authors were trying to establish which metabolites separate the patients before and after and the controls before treatment. This post will present Panel 5E which summarizes the methyl transferring pathways that predominate the data

This post is going to skip presenting Panel 5D that presented amino acid metabolite changes that tended to be around 2x the baseline values. We may need to come back to it when there’s a better understanding of the salvage pathways in yeast and humans in telomeres.

Fig. 6 Pathways

The reviewing this section note will be made when the pathways intersect that of copper and copper cofactor enzymes

This will be an awkward transition from the salvage pathway, and niacin supplementation to the salvage pathway in telomeres in yeast. Three references were found from the JS Smith Lab that increased my understanding of the role of niacin in telomeres.

NAD⁺ and telomeres

The Sir2 enzyme is an NAD-dependent protein deacetylase that is required for turning off the silent mating-type loci, telomeres, and the ribosomal DNA (rDNA). Mutations in the NAD salvage gene NPT1 weaken all three forms of silencing and also cause a reduction in the intracellular NAD level. [1]

What are telomeres and telomerase?

Telomerase is a ribonucleoprotein that adds a species-specific telomere repeating sequences to the 3′ end of telomeres. A telomere is a region of repetitive sequences at each end of the chromosomes of most eukaryotes that function to prevent fusion with other chromosomes and DNA damage.

In higher eukaryotes, telomere length plays a major role in regulating cellular senescence. This is especially true in primary fibroblasts that do not express telomerase. Telomeres become shorter during each cell division. The Hayflick limit is reached when telomeres become short enough to senesce.

Yeast telomerase is constantly expressed with no progressive shortening in mitotic daughter cells. Mutations in telemerase can mimic mammalian aging with progressive shortening of telomeres. Telomerase deficient est2Δ rad52Δ mutants can’t repair their telomeres. Deleting SIR2 further accelerates senescence within the population. [3]

In yeast the lack of Sir2 results loss of telomeric silencing, which increases the transcription of non-coding TElomeric Repeat containing RNAs (TERRA) Wikipedia has an excellent page on these controllers of telomerase activity. Telomeric silencing refers to the observation that protein coding genes located close to telomers tend to be transcribed less.

A mammalian telomere, NAD⁺ comes in here

1. Rap1p binds to the telomeric repeats.
2. Sir3p/Sir4p/Sir2p complex is recruited to Rap1p. Enter NAD⁺
3. Sir2p deacetylates histones in the adjacent nucleosome.
4. Deacetylated histones recruit more Sir3p/Sir4p/Sir2p complex. Enter NAD⁺
5. The cascade of deacetylation spreads away from the telomeres.
6. Spreading of deacetylation is countered by Histone Acetyl Transferases (HATs)

Back to yeast telomeres and NAD⁺ salvage

Transcriptional repression can be highly localized and transient, such as at the promoters of specific genes, or more widespread across large regions of the genome that remain in a repressive and condensed state for extended periods. These latter domains tend to be heterochromatic and stable, sometimes even through multiple generations. In budding yeast, HML, HMR, and telomeres are generally considered to be the heterochromatin equivalents in this organism. [3]

The authors discussed nutrient sensing ensembles of proteins that also control aging. These include sensors of mitochondrial function that is not just NAD⁺ production but also the ADP to ATP ratio. This is where we think that Cu⁺ and mitochondrial cytochrome C oxidase may synergize with agents that boost NAD⁺.

yeast telomere references

1. Sandmeier JJ, Celic I, Boeke JD, Smith JS. Telomeric and rDNA silencing in Saccharomyces cerevisiae are dependent on a nuclear NAD(+) salvage pathway. Genetics. 2002 Mar;160(3):877-89. PMC free article
2. Gallo CM, Smith DL Jr, Smith JS. Nicotinamide clearance by Pnc1 directly regulates Sir2-mediated silencing and longevity. Mol Cell Biol. 2004 Feb;24(3):1301-12. PMC free article
3. Wierman MB, Smith JS. Yeast sirtuins and the regulation of aging. FEMS Yeast Res. 2014 Feb;14(1):73-88. PMC free article

We had thoughts, at one time, of measuring telomere length in cultured fibroblasts as a index of aging. Why not cultured hair follicles since so many customers report restoration of pigmentation in their hair when consuming BioCu1. Hair follicles are also more mitochondria reliant for their daily activities… unless the aging process is taking its toll. The more we thought about the proper controls, the harder and more expensive things seemed. Perhaps we are over thinking the story.

1. Background Cu1 vs Cu2, don’t forget niacin
2. Oxidative stress and telomere length
3. Exposure to Cu1 and Cu2
4. Aging studies: hair follicles in culture
   1. hair follicles are complicated organs
   2. Reactive oxygen species (ROS) not defined
5. Follicles from hairs that have turned dark

Background Cu1 vs Cu2, don’t forget niacin

• Ctr1 describes the Ctr1 Cu(I) transport channel. Cu(II) does not get transported until it is reduced to Cu(I). We have always assumed that Cu(I)niacin is more bioavailable than Cu(II) sulfate.
• Copper chaperones deliver Cu in the +1 oxidation state to their target enzymes. This is why we always thought, but never proved, that our copper is better.
• Niacin in the Covid describes how we began to realize that Cu(I) and niacin likely work together.
• Fatty Liver in Cows expands on the interplay between niacin and copper.

Oxidative stress and telomere length

von Zglinicki T. Oxidative stress shortens telomeres. Trends Biochem Sci. 2002; 27: 339-344 Sci-Hub free paper

Some images were imported of the position of telomeres in a chromosome and examples of telomere shortening with cell divisions. Stem cells, red, keep a fairly constant telomere length. The moment a cell differentiates into a somatic cell (green) with a designated function, telomeres start to shorten. The Hayflick limit (HL) is the number of times a cell may divide before division stops. Crises (blue) may alter gene expression and increase telomere length. PD is the population doubling. (b) The downward green arrows in the von Zglinicki Fig 1 indicates that stress can shorten telomere length more rapidly than normal population doubling. (c) High stress and/or lack of good oxidative defenses, indicated by the width of the blue triangle, can send the telomere length into an almost vertical downward spiral. Good oxidative stress defense can send the telomere length response to

Though this review was published over 20 years ago, it was very insightful and extremely useful for generating hypotheses. The Cu(I) of BioCu1 supplies Cu/Zn superoxide dismutase with a necessary cofactor. Niacin can be a precursor of NADH and NADPH, a cofactor for many anti-oxidant enzymes.

Exposure to Cu1 and Cu2

These are some thoughts on how we might design a cell culture experiment in fibroblasts or maybe even hair follicles in culture. Fibroblasts from individual patients and hair follicles could be studied.

1. For starters we will need the culture medium without red dye that is used to indicate pH. We will add the same molar amount of Cu1 used in previous cell culture studies to the culture medium. We will take UV/Vis spectrophotometer scans every one minute for 15 minutes. We will calculate the time window in which we can keep the Cu1 in culture medium on the cells with less than 10% being oxidized to Cu2. (Cupric copper is blue).
2. We could add both coppers to cultured follicles and/or fibroblasts for the designated time.
3. After the incubation period, the copper containing media could be moved and the copper adhering to the outsides of the cells rinsed away.
4. As another control for the Cu1 , we might want to consider the same molar amount of niacin without the copper. As a control for the cupric sulfate we might want to add a non copper sulfate.
5. Do we have a way of measuring the copper that was taken up by the cells?
6. The next step might be to measure ATP content.

This is already starting to seem like a considerable amount to perform fibroblast or cultured hair follicle experiments. Others are using hair follicles in culture to study telomere aging.

Aging studies: hair follicles in culture

Stone RC, Aviv A, Paus R. Telomere Dynamics and Telomerase in the Biology of Hair Follicles and their Stem Cells as a Model for Aging Research. J Invest Dermatol. 2021 Apr;141(4S):1031-1040. free article

• This review proposed that telomere length dynamics play an important role in the biology of the hair follicle (HF),
• HF are mini organs that show an intriguing aging pattern in humans.
• The pigment producing unit ages quickly but epithelial stem cells (ESC) are more aging resistant. Telomerase deficient mice with short telomeres display an aging phenotype of hair graying and hair loss that is attributed to impaired HF ESC mobilization.

hair follicles are complicated organs

Reactive oxygen species (ROS) not defined

The lay reader, and even trained biologists, would have to do so much background reading to properly understand what is being said in this review. This review does not discuss the anti-oxidant enzyme Cu/Zn super oxide dismutase simply because no one can think of everything at once.

This is all very interesting, but we’ve got to

Follicles from hairs that have turned dark

We have so many anecdotal stories of customers who have had their original hair color return after taking BioCu1. Why not just analyze the follicles from such hairs that have turned dark and compare them with hair follicles from the same customer before BioCu1.

For an overview of gray hair and a few works on the mitochondria requirement to grow hair, visit the gray hair post. The starting point of this post is Women’s World 8 essential oils for hair growth. The strategy is to do a PubMed search on each and then figure out which phytochemicals are in each and what their receptors are.

Rosemary oil

It took six months to see a difference in androgen associated hair loss. [1]

1. Panahi Y, Taghizadeh M, Marzony ET, Sahebkar A. Rosemary oil vs minoxidil 2% for the treatment of androgenetic alopecia: a randomized comparative trial. Skinmed. 2015 Jan-Feb;13(1):15-21. PMID: 25842469. PubMed

pumpkin seed oil

In a 3 month clinical trial PSO showed promise in treating female pattern hair loss. In this 24 week clinical trial with men with androgen alopecia Mean hair count increases of 40% were observed in PSO-treated men at 24 weeks, whereas increases of 10% were observed in placebo-treated men (P < 0.001). Adverse effects were not different in the two groups.

1. Ibrahim IM, Hasan MS, Elsabaa KI, Elsaie ML. Pumpkin seed oil vs. minoxidil 5% topical foam for the treatment of female pattern hair loss: A randomized comparative trial. J Cosmet Dermatol. 2021 Sep;20(9):2867-2873 PubMed
2. Cho YH, Lee SY, Jeong DW, Choi EJ, Kim YJ, Lee JG, Yi YH, Cha HS. Effect of pumpkin seed oil on hair growth in men with androgenetic alopecia: a randomized, double-blind, placebo-controlled trial. Evid Based Complement Alternat Med. 2014;2014:549721. PMC free article
3. Teeranachaideekul V, Parichatikanond W, Junyaprasert VB, Morakul B. Pumpkin Seed Oil-Loaded Niosomes for Topical Application: 5α-Reductase Inhibitory, Anti-Inflammatory, and In Vivo Anti-Hair Loss Effects. Pharmaceuticals (Basel). 2022 Jul 27;15(8):930. PMC free article

peppermint oil

Only one abstract was found on PubMed which focused on a type of hair loss specific to African American women which stated that there was not great evidence that this substance was effective.

Ezekwe N, King M, Hollinger JC. The Use of Natural Ingredients in the Treatment of Alopecias with an Emphasis on Central Centrifugal Cicatricial Alopecia: A Systematic Review. J Clin Aesthet Dermatol. 2020 Aug;13(8):23-27. Epub 2020 Aug 1. PubMed

lavender oil

Also see the thyme oil section for the Turkish combination study.

clary sage oil

I could find nothing on PubMed concerning clary sage oil and hair loss. Titles indicate it is calming.

cedarwood oil

Also see the thyme oil section for the Turkish combination study.

tea tree oil

A study out of Saudi Arabia tested the hypothesis that adding tea tree oil would improve hair growth in male androgenic alopecia. These authors used a multimodal microemulsion of minoxidil (a dihydrotestosterone antagonist), diclofenac (a nonsteroidal anti-inflammatory agent), and tea tree oil which they viewed as an anti-infective agent. This publication reports stability factors that might be of interest to Mito. The good news is that significant hair growth was recorded after 32 weeks of use compared to the placebo and traditional medication.

Farouk Sakr F, Gado A, Mohammed H, Ismail AAN. Preparation and evaluation of a multimodal minoxidil microemulsion versus minoxidil alone in the treatment of androgenic alopecia of mixed etiology: a pilot study. Drug Des Devel Ther. 2013;7:413. [PMC free article]

thyme oil

The Ezekwe review is the only thing that showed up on PubMed {1] This is not to say the research has not been performed. The Ezekwe review lead to a Turkish study of a combination of essential oils that are to be rubbed into the scalp. [2] The active group in this clinical trial received a compound available in Turkey: Revigen® Areata solution (Mikro-Gen Co, Istanbul, Turkey) which has essential oils, Thyme vulgaris, Lavandula agustifolia, Rosmainus officialis and Cedrus atlantica and mixed in carrier oil which was a combination of jojoba and grape seed oils.

1. Ezekwe N, King M, Hollinger JC. The Use of Natural Ingredients in the Treatment of Alopecias with an Emphasis on Central Centrifugal Cicatricial Alopecia: A Systematic Review. J Clin Aesthet Dermatol. 2020 Aug;13(8):23-27. Epub 2020 Aug 1. PMC free article
2. Özmen I, Çalişkan E, Arca E et al. Efficacy of aromatherapy in the treatment of localized alopecia areata: a double-blind placebo controlled study. Gulhane Med J. 2015;57:3. free article

If the particular source of thyme oil contains the terpene thymol it might be effective as a skin permeability enhancer to get active ingredients past the stratum corneum.

Holistic approach: biotin, niacin,more

A very recent review mentioned popular use of biotin in products for hair regrowth but seemed to be much more optimistic about niacin. [1] The corresponding author of this paper is Raja Sivamani MD, who usually publishes with four or five institutes as his home base. These include a medical school and several research centers. Raja Sivamani has been part of numerous clinical trials.

College of Medicine, California Northstate University, 9700 W Taron Dr, Elk Grove, CA 957
Integrative Skin Science and Research, 1495 River Park Drive, Sacramento, CA
Pacific Skin Institute, 1495 River Park Dr Suite 200, Sacramento, CA
Department of Dermatology, University of California-Davis, Sacramento, CA

Here is what the review [1] has to say about B vitamins:

B Vitamins

B vitamins, including niacin (vitamin B3), biotin (vitamin B7), and folic acid (vitamin B9) have been implicated in hair loss. For example, in addition to the well-documented pellagra characteristic of niacin deficiency, alopecia is an additional common clinical finding associated with deficient niacin [43]. However, no studies have directly assessed niacin levels among patients presenting solely with hair loss, and studies have found no significant difference in folate levels between alopecia patients and control subjects [57,58].

“Biotin, a cofactor for carboxylation enzymes with dietary sources including protein, has been more extensively assessed for effects on hair parameters, and it is included in a variety of supplements or serums intended for hair health [44]. Genetic biotin deficiency is associated with severe dermatitis and alopecia (infantile) and sparse or absent scalp, eyebrows, and eyelash hair (infantile). Similarly, acquired biotin deficiency is characterized by alopecia and brittle nails [43]. A 2016 study assessed serum biotin levels in women with self-reported hair loss and found 38% of patients reported a biotin deficiency [59]. However, this study did not include matched controls.

Despite many misconceptions, biotin functions to increase hair strength, rather than hair growth. Furthermore, biotin can interfere with troponin and thyroid testing. For example, excess serum biotin can result in a falsely low TSH level [60], and unnecessary supplementation can lead to missed cardiac events [61]. Yet, biotin supplement use has depicted increasing trends from 1999 to 2016; a cross-sectional survey study found self-reported use of biotin at 1 mg/d or greater to increase from 0.1% (95% CI 0.0–0.05%) in 1999 to 2.8% (95% CI 1.9–3.9%) in 2015–2016 [61].

Despite biotin’s popular inclusion in marketed hair supplements, there is no indication that biotin supplementation should be used among healthy individuals [43]. While biotin supplementation has shown benefit specifically among cases in which acquired or inherited causes of biotin deficiency are identified, there is insufficient evidence supporting supplementation among healthy individuals who are not deficient [62]. Thus, vitamin B testing may only be clinically useful in cases of suspected biotin deficiency, where biotin supplementation may improve the clinical condition.”

3.2.3. Light-Based Approaches “Low level light therapy refers to therapeutic exposure to low levels of red and near infrared light [118]. Studies have demonstrated increased hair growth in mice with chemotherapy-induced alopecia and AA, in addition to both men and women human subjects. Proposed mechanisms of efficacy include stimulation of epidermal stem cells residing in the hair follicle bulge and promoting increased telogen to anagen phase transition [119].” Note: this is near infrared light that I’ve written about in another post several years ago because I thought there’s be synergy with cuprous niacin. Red Light Post.

Section 3.3 on platelet rich plasma might be a good thing to discuss with Charlie Barker’s neighbor at the Flagstaff incubator.

1. Natarelli N, Gahoonia N, Sivamani RK. Integrative and Mechanistic Approach to the Hair Growth Cycle and Hair Loss. J Clin Med. 2023 Jan 23;12(3):893. PMC free article

The polyol pathway is a two step process that converts glucose to fructose. This post explores how copper deficiency may make intermediates in this pathway worse.These findings have relevance to Alzheimer’s Disease.

Xu J, Begley P, Church SJ, Patassini S, McHarg S, Kureishy N, Hollywood KA, Waldvogel HJ, Liu H, Zhang S, Lin W, Herholz K, Turner C, Synek BJ, Curtis MA, Rivers-Auty J, Lawrence CB, Kellett KA, Hooper NM, Vardy ER, Wu D, Unwin RD, Faull RL, Dowsey AW, Cooper GJ. Elevation of brain glucose and polyol-pathway intermediates with accompanying brain-copper deficiency in patients with Alzheimer’s disease: metabolic basis for dementia. Sci Rep. 2016 Jun 9;6:27524. PMC free article

Introduction …. to Alzheimer’s Disease

“Alzheimer’s disease” (AD) may occur through interplay of environmental, genetic, and metabolic factors according to these authors. The authors list some pathological manifestations:

• Aβ-amyloid deposition
• neurofibrillary tangles comprising tau protein
• impaired cerebral glucose metabolism (‘hypo metabolism’) with insulin resistance and defective carbohydrate regulation
• oxidative stress and inflammation
• cerebrovascular amyloid angiopathy (CAA)
• enhanced advanced glycation end-product (AGE) formation
• defective copper regulation.
• defects in glucose uptake

The polyol pathway

The polyol pathway is a means of taking the six carbon sugar glucose with the aldehyde C=O group on the top of the structure to a ketone form, fructose, with the C=O on the second carbon from the top. In this way, the propensity to form advanced glycation end products is reduced.

Copper should theoretically increase the utilization of glucose if copper is a pivotal player in brain glucose utilization simply by keeping the electron transport chain running.

The authors analyzed post-mortem tissue from seven brain regions of patients with AD and controls who did not have clinical evidence of dementia. The possibility exists that one or more patients had undiagnosed mild T2D or pre-diabetes. The authors reported that elevated glucose levels have previously been linked to altered tissue-copper regulation in the context of diabetes.

Regions most prone to damage in AD

• hippocampus
• entorhinal cortex
• middle temporal gyrus

Three regions are less effected

• cingulate gyrus,
• sensory cortex
• motor cortex

The cerebellum is largely spared.

The inverse relationship between glucose and copper

Free glucose is elevated in AD brain, particularly in vulnerable regions where elevated glucose may well be linked to copper deficiency and tissue-damage.

• Overall mean brain-glucose levels were higher (P = 9.7 × 10^−13) in AD vs controls
• Brain copper values lower (P = 1.6 × 10^−8) in cases vs controls.

P values that the AD is different from the controls is written below the data points. There was modest evidence that overall brain-copper and brain-glucose values were inversely correlated (P = 0.021) in AD-patients but not controls

Brain-sorbitol and brain-fructose levels are also elevated in AD brain, consistent with increased polyol pathway flux. Defects may be in glucose utilization, the TCA cycle, or the electron transport chain.

The six carbon sugar pile up in AD and T2D

A further, mechanistically-related finding is that pan-cerebral brain-copper deficiency accompanies these glucose and polyol-pathway defects: elevated tissue-CML levels provide a molecular linkage between elevated brain glucose and copper deficiency.

Impaired neuronal glucose utilization could play a major role in tissue-damage in AD, possibly via defective mitochondrial metabolism caused by deficient copper supply to cytochrome C oxidase subunits I and II (COI and COII), leading to impaired function of cytochrome c oxidase. These findings provide a clear molecular linkage between mechanisms of tissue damage in AD and T2D.

These “fold change” plots compare controls and disease in tissue from (a) each of seven brain regions from patients with AD (n = 9) and controls (n = 9) and (b) whole brain homogenates from diabetic (n = 7) and control (n = 7) rats.

1. Complicated methods of analysis are not presented in this post.
2. Solid lines are average fold changes.
3. Dashed lines are the 95% Confidence intervals.
4. The dark line at “1” represents a fold change of 1, that is “no change”

The process by which elevated brain glucose is linked to defective cell-copper uptake and intracellular transport to key copper proteins such as COI, COII, SOD1 and SOD3, may well provide a new target for diagnostic imaging or therapeutic intervention.

A few words of extreme caution

The same authors performed a followup study in post mortem brains of type 2 diabetes patients. Transition metal levels were measured, but glucose, sorbitol, and fructose were not. In contrast to AD patients, copper was found to be elevated in the hippocampus of T2D patients.

Philbert SA, Schönberger SJ, Xu J, Church SJ, Unwin RD, Cooper GJS. Elevated hippocampal copper in cases of type 2 diabetes. EBioMedicine. 2022 Dec;86:104317. PMC free article

This is a direct quote from the discussion of the data

“Despite copper’s key roles in some antioxidant processes (e.g., those catalysed by superoxide dismutase 1 and by complex IV), it can also participate in the production of harmful reactive oxygen species (ROS) via the Fenton and Haber–Weiss reactions. There is evidence that Cu^II can bind to advanced glycation end-products (AGEs) whilst retaining its redox–active properties. As protein glycation is significantly increased in T2D due to hyperglycaemia and polyol-pathway activation, the elevated hippocampal copper measured in the present study has the potential to cause increased AGE-Cu^II complex formation. The formation of AGE-Cu^II complexes could thereby lead to increased Cu^II-mediated ROS production, and decreased copper bioavailability for antioxidative pathways due to the selective binding of extracellular Cu^II with AGEs. However, Cu^II is less abundant than Cu^I, so the role that AGE-Cu^II complexes might play in the pathogenesis of T2D remains to be clarified.”

Stay tuned for more on these AGE-Cu^II complexes.