Zinc Deficiency and elevated IL-6 > mast cell degranulation>histamine excess; also >microbiome dysbiosis.

Confounding variable, Genes Table Section G. / Product request - zinc nanoparticle tapioca & pomegranate peel-inner pith gummies - boost the butyrate species

G) Confounding Variable – Zinc deficiency as a comorbid condition would exacerbate the risk of elevated homocysteine due to the role of zinc ion binding for the BHMT, SHMT, and MTR proteins, and lead to elevated IL-6 due to epigenetic changes; IL-6 degranulates mast cells, adding to histamine excess.

• The one-carbon methylation cycle uses zinc in the BHMT, SHMT, and MTR enzymes and deficiency of zinc can lead to reduced methylation of the IL-6 promoter which leads to increased IL-6 production, and mast cell degranulation.

• Zinc Deficiency can lead to elevated IL-6 in an age associated risk for older adults.

• IL-6 degranulates mast cells and would add to histamine excess.

• Zinc deficiency leads to lower IL-6 promoter methylation which leads to more production of IL-6 in elderly subjects. Younger subjects had a reverse response – hypermethylation with zinc deficiency.

Addition - someone asked for more of an explainer. Zinc deficiency is bad for health. It is a frequent underlying undiagnosed problem, common in older adults who may need about double the current RDA for maintaining thymus function (*as in the recommendation seems to be wrong - just bluntly). If this wasn’t screened for in any study looking at inflammation, it would skew the results. And **** if the screening is based on ‘the recommended guideline’ then the result might indicate that the older adult has adequate zinc, while instead the older adult may have reduced thymus function, reduced DNA methylation and aberrant protein production - a lack of important proteins. Zinc is needed in three of the enzymes used in the one-carbon methylation cycle/s and it is needed in Zinc finger proteins to be able to transcribe a significant number of other proteins.

In this article the effect of zinc deficiency on the one-carbon methylation cycle is being discussed. It disrupts it in a way that the one-carbon cycles no longer work in the cell nucleus and they are needed for us to be able to methylate DNA for epigenetic control. Too little or too much methylation can affect whether the gene will be transcribed or not, like an on/off switch or a cap on a tube of toothpaste - add the cap, remove the cap, add the cap, remove the cap. Hundreds of genes are turned on and off in that way each and every 24-hour cycle - if we have a healthy and functioning circadian cycle and well-nourished and functioning one-carbon methylation cycles.

The lack of zinc leading to lack of DNA methylation leads to age related effect of increased production of the inflammatory cytokine IL-6 (which can be elevated in COVID19). IL-6 degranulates mast cells, which adds to histamine excess risk and releases more inflammatory cytokines. The zinc deficiency, particularly in older people, may be a causal factor in a cytokine storm/hyperinflammation which also has histamine excess symptoms and risks if it is a chronic problem that remains undiagnosed and untreated with adequate zinc supplementation or very good food sources. One aspect of the research I share in the later discussion that is unclear is what the hypermethylation of the IL-6 promoter does for the younger mice or human cells - this is not at clinical trial stages yet. It does help explain why clinical trials for higher dose zinc supplementation is needed for older adults with chronic inflammatory conditions. It does help explain why zinc deficiency is associated with inflammation. It would suggest that zinc deficiency should be considered when histamine excess is present.

If the IL-6 promotion occurs in older adult human cells, it may also occur in specific conditions in younger adults too. Again, more research is needed, and in the meantime - zinc, in balance with copper, is critical to health. *Hypermethylation tends to silence genes and sometimes permanently and the change may be passed to the next generation. (Ehrlich, 2019) Zinc deficiency leading to hypermethylation of the IL-6 gene might leave the younger person more susceptible to an infection where some cytokine response is needed.

RE food sources of zinc: Oysters are an exceptional source and could reach toxic amounts if eaten as a daily food. Meats, fish, are good sources, and pumpkin seeds, other beans and seeds to a lesser extent than meats. Zinc can be difficult to get enough of in a vegetarian diet. Some people may have an excess copper problem that leads to poor absorption of zinc. The galvanized zinc water pitcher at the end of the post is just an extra, not really intended as a dietary recommendation, more of a caution not to heat tomato sauce in iron or old galvanized zinc containers.

Several One-carbon Cycle/s enzymes require zinc.

The BHMT protein is portrayed in Li, F., et al., 2008, Figure 3, with atoms of zinc in place. Figure 1 shows the interconnected pathways of the methylation of homocysteine to methionine and folate to 5-THF. The SHMT, MTHFR, MTR, and BHMT enzymes have critical roles in the double pathway that removes a methyl group from betaine and adds it to homocysteine for methionine production or to a folate metabolite for 5-HTF production. Dimethglycine is produced from betaine in the process. Limit zinc and the double cycle doesn’t work as BHMT and SHMT both use zinc. (Li, F., et al., 2008, Figure 1, Figure 3) Lack of 5-HTF limits methylation of DNA for epigenetic control.

MTR, methionine synthase, is also a zinc-dependent methyltransferase. Reduced expression of Methionine synthase (MS) was observed during zinc deficiency in an animal-based study. Homocysteine was elevated 65% in serum concentrations with no significant difference in serum folate levels seen between the zinc deficiency diet group and the pair fed group. Intake and weight were decreased in the group with the zinc deficient diet but not significantly more than the pair fed. (Jing, et al., 2015)

Age related risk - older adults need about double the zinc as the current RDA, for maintaining thymus function.

Zinc deficiency seems to have an age-related risk for elevated IL-6. The elderly, especially those with SHMT, MTHFR, MTR, and/or BHMT alleles that cause dysfunction, may have increased IL-6 when zinc deficient due to reduced IL-6 promoter methylation, (Azimi, et al, 2022) – and IL-6 increases mast cell activity and histamine production.

IL-6 degranulates mast cells.

IL-6 is a pleitropic cytokine which is a mediator for mast cell maturation and upregulates production of histamine rather than promoting storage. (Conti, et al., 2002) IL-6 promoted mast cell maturation, proliferation and increased reactivity by down regulating the inhibitory SOCS3 pathway. Expression of SOCS3 was reduced and it correlated with methylation of the SOC3 promoter. The levels of sIL-6R were suppressed and reduced levels of sIL-6R have been observed in patients with mastocytosis. IL-6 blockade is suggested by Desai, et al., 2016. *See Treatment Options below.

Zinc deficiency > less IL-6 promoter methylation > more production of IL-6 in elderly subjects. Younger subjects (mice or human cells) > hypermethylation with zinc deficiency.

Azimi, et al, found that “…the spleens of older mice fed a zinc-deficient diet revealed lower IL-6 promoter methylation, which was associated with an increase in IL-6 and intercellular adhesion molecule 1 (ICAM1) expression, but these changes were not observed in younger mice [19]. This effect of zinc-deficient diet on IL-6 expression in young and old rodent cells contrasts with the effect seen in young and old human cells. […]

In another study, low zinc concentration had a heterogeneous effect on the methylation of interleukin 6 (IL-6) in lymphoblastoid cell lines and the human monocytic cell line THP-1 from both aged and young subjects [19]. In this study, cells from elderly subjects in zinc-deficient culture media exhibited significantly lower IL-6 promoter methylation. Interestingly cells from younger subjects exhibited hypermethylation on the IL-6 promoter. Global DNA methylation status in both groups remained unchanged.” (CC BY-NC-ND Azimi, et al, 2022)

Medical history regarding zinc and B1 and B6 deficiency, exacerbated emotional dysregulation and some schizophrenia like symptoms:

At a peak of early negative mental changes, around age 48, the author learned of pyroluria and added 50 mg zinc per day and 250 mg B6 or 100 mg Pyridoxal 5’-Phosphate twice a day. At the time dissociative symptoms were significant and function was difficult. There were significant white spots on the fingernails, which can be indicative of zinc deficiency. Around that time thiamin deficiency came to the author’s notice and 300 mg of thiamin improved a faint irregular heartrate and disoriented thinking within a day or too. Severe anorexia was a problem at the time, likely related to the severe zinc deficiency and undiagnosed hyperthyroidism. The author needs to continue the B6, P5P, and zinc on an almost daily basis or mental symptoms start returning: anxiety, agitation. (Age at the time of writing – 56.) [Personal experience of the author.]

Treatment options, zinc and pomegranate products.

• Pomegranate peel extract is an inhibitor of IL-6. (Čolić, et al, 2022; Elkady, et al., 2021, Figure 3; Mastrogiovanni, et al., 2019; Qabaha, et al., 2019, Figure 1; Sayed, et al, 2022)

• Decreased thymus function with age can be improved with extra supplementation of zinc. Thymus function tends to be reduced in the elderly and can be improved with supplementation at higher than current guidelines. Older adults likely require about twice as much zinc as younger and middle-aged adults to support or restore thymus function. (Kodama, et al, 2020)

Product development request: Pomegranate peel can be used to make nanoparticles with silver or gold for antimicrobial benefits (Monira, et al, 2017; Nadagouda, et al, 2014; Nasiriboroumand, et al, 2018; Rao, et al, 2013; Yang, H., et al, 2016) and zinc nanoparticles made with lavender were found to be safe and effective against all phases of Toxoplasmosis gondii infection. (Saadatmand, et al, 2021) Nanoparticles are more bioavailable or biotoxic because it is difficult for the body to remove them. Pomegranate peel has adsorbent properties and binds with +2 and other ions.

Pomegranate/peel phytonutrients can help clump toxic nanoparticles (from smog or other sources) into a larger cluster which the white blood cells are able to identify as something to remove. Nanoparticles are a risk, or a more effective supplement/medication because they are too small for the body’s normal clean-up mechanisms.

Or . . . zinc nanoparticle tapioca gummies. Adding pomegranate inner pith, and zinc or zinc pom peel nanoparticles, in a tapioca chewable gummy would make it a resistant starch prebiotic for butyrate species. The zinc, pith and tapioca promote butyrate species of the microbiome. Advantage – all ages friendly, Seniors (Kodama, et al, 2020) and adults could have a larger number listed as a daily serving and one for children.

MicroRNA are involved in the many varied benefits seen with pomegranate peel extract (the fruit is rich in nutrients, but the peel is more concentrated):

Pomegranate phytonutrients and others promote health by promoting miRNA that up and down regulate genes in beneficial ways due to the control of mRNA by microRNA (miRNA).

Milenkovic, et al., analyzed the effects of nine dietary polyphenols on apolipoprotein E (Apo E) deficient mice which had altered lipid metabolism and control group mice. The polyphenols were tested separately and changes in miRNA were measured. Interestingly the baseline differences in microRNA between the Apo E deficient mice and control group were found to be somewhat normalized by the polyphenol treatment. Many genes that were down or up regulated differently in the Apo E -/- mice, than the control mice, were improved more toward normal function with polyphenol treatments. All of the tested polyphenols were found to affect five of the same microRNA. The biological pathways modulated by the five miRNAs were then identified and thirty were also being affected by the modulation of mRNA. (Milenkovic, et al., 2012)

What were the tested polyphenols? Drumroll… the phenolic acids: caffeic and ferulic acid; and the flavonoids: quercetin, anthocyanin, catechin, proanthocyanin, hesperidin, and naringenin; and curcumin. The animals were supplemented with one of the polyphenols for a two-week time period. (Milenkovic, et al., 2012)

• *This section was expanded on in the earlier post: (1) microRNA are the real regulators of gene transcription. (substack.com)

• This post is about dysfunction in the methylation cycles within a cell’s cytoplasm having negative impact on the cell nucleus being able to methylate DNA properly or at all. (3) Nature loves a good design; Cancer, POTS, Epigenetics & the One-Carbon Methylation Cycles. (substack.com)

Other posts related to the Genes Table series:

• (2) “microRNA, elevated homocysteine and is there a role for excess Retinoic Acid?” (substack.com) *The hyperhomocysteinemia/homocystinuria section of my Genes Table.

• “Pollutants in Human Plasma Found via Double-Filtration Plasmapheresis Plasma Exchange”, James Lyons-Weiler. (substack.com) *This is a cross post. Heavy metal toxins and thread like objects and aluminum-silicon combinations were found in a filtration of plasma procedure. This is related to pomegranate because the peel is effective at clumping nanoparticles into larger clumps that are big enough for white blood cells to sense as something needing to be removed.

• (.5) “POTS - Postural Orthostatic Tachycardia Syndrome, can be epigenetic & therefore may be reversible”. (substack.com) *The Dystonia section of my Genes Table.

*Eventually I plan to get this all into more of a chapter/book format or webpages in a drop-down menu format, from less info to more detailed pages. Pick what level of info is desired.

Galvanized Zinc tools and washtubs were once commonly used. Photo by Clément Falize on Unsplash

Water and Zinc - (LennTech.com) a nice overview about zinc in the environment, water, and diet.

A galvanized zinc water pitcher would be unlikely to add much zinc to the water, but if acidic lemonade was put in it and left out on a hot sunny day, the heat and acidity might add zinc content to the water. Cooking acidic tomato sauce in an old galvanized tub may cause too much solubility of zinc. Very alkaline, above 11 pH also would cause increased solubility of the zinc from a galvanized zinc container. (LennTech.com)

Natural water supplies do have some zinc. Excess zinc is a health risk. (LennTech.com)

Disclaimer: This information is being shared for educational purposes within the guidelines of Fair Use and is not intended to provide individual health guidance.

Reference List

(Azimi, et al, 2022) Azimi, Z., Isa, M.R., Khan, J., Wang, S.M., Ismail, Z., (2022). ‘Association of zinc level with DNA methylation and its consequences: A systematic review’. Heliyon. Sep 28;8(10):e10815. doi: 10.1016/j.heliyon.2022.e10815. PMID: 36203899; PMCID: PMC9530842. Available at: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9530842/ (Accessed: 22 Nov 2022)

(Čolić, et al, 2022) Čolić M, Bekić M, Tomić S, et al., (2022). ‘Immunomodulatory Properties of Pomegranate Peel Extract in a Model of Human Peripheral Blood Mononuclear Cell Culture. Pharmaceutics’. May 27;14(6):1140. doi: 10.3390/pharmaceutics14061140. PMID: 35745713; PMCID: PMC9228601. Available at: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9228601/  (Accessed: 21 October 2022)

(Conti, et al., 2002) Conti, P., Kempuraj, D., Di Gioacchino, M., Boucher, W., Letourneau, R., Kandere, K., et al., (2002). ‘Interleukin-6 and mast cells’. Allergy Asthma Proc. Sep-Oct;23(5):331-5. PMID: 12476543. Available at: https://pubmed.ncbi.nlm.nih.gov/12476543/ (Accessed: 22 Nov 2022)

(Desai, et al., 2016) Desai, A., Jung, M.Y., Olivera, A., Gilfillan, A.M., Prussin, C., Kirshenbaum, A.S., et al., (2016). ‘IL-6 promotes an increase in human mast cell numbers and reactivity through suppression of suppressor of cytokine signaling 3’. J Allergy Clin Immunol. Jun;137(6):1863-1871.e6. doi: 10.1016/j.jaci.2015.09.059. Epub 2016 Jan 7. PMID: 26774658; PMCID: PMC4899186. Available at: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4899186/ (Accessed: 22 Nov 2022)

(Elkady, et al., 2021, Figure 3) Elkady, A., Abdel-Rahman, I., Fouad, S., Allemailem, K., Istivan, T., Ahmed, S., Hasan, A., Osman, H., Elshabrawy, H., (2021). Pomegranate Peel Extract Is a Potential Alternative Therapeutic for Giardiasis. Antibiotics. 10. 705. 10.3390/antibiotics10060705. Available at: https://www.researchgate.net/publication/352340267_Pomegranate_Peel_Extract_Is_a_Potential_Alternative_Therapeutic_for_Giardiasis Figure 3: https://www.researchgate.net/figure/Pomegranate-extract-reduced-levels-of-IL-6-TNF-a-and-increased-NO-production-in-animals_fig1_352340267 (Accessed: 22 Nov 2022)

(Ehrlich, 2019) Ehrlich, M., (2019). DNA hypermethylation in disease: mechanisms and clinical relevance. Epigenetics. Dec;14(12):1141-1163. doi: 10.1080/15592294.2019.1638701. Epub 2019 Jul 8. PMID: 31284823; PMCID: PMC6791695. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6791695/

(Jing, et al., 2015) Jing, M., Rech, L., Wu, Y., Goltz, D., Taylor, C.G., House, J.D., (2015). ‘Effects of zinc deficiency and zinc supplementation on homocysteine levels and related enzyme expression in rats’. J Trace Elem Med Biol. Apr;30:77-82. doi: 10.1016/j.jtemb.2014.10.013. Epub 2014 Nov 13. PMID: 25467853. Available at: https://pubmed.ncbi.nlm.nih.gov/25467853/ (Accessed: 23 Nov 2022)

(Kodama, et al, 2020) Kodama, H., Tanaka, M., Naito, Y., Katayama, K., Moriyama, M., (2020). ‘Japan’s Practical Guidelines for Zinc Deficiency with a Particular Focus on Taste Disorders, Inflammatory Bowel Disease, and Liver Cirrhosis’. Int J Mol Sci. 21(8):2941. Published 2020 Apr 22. doi:10.3390/ijms21082941 Available at: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7215354/ (Accessed: 21 October 2022)

(Li, F., et al., 2008, Figure 1, Figure 3) Li, F., Feng, Q., Lee, C., et al., (2008). ‘Human betaine-homocysteine methyltransferase (BHMT) and BHMT2: common gene sequence variation and functional characterization’. Molecular Genetics and Metabolism. Jul;94(3):326-335. DOI: 10.1016/j.ymgme.2008.03.013. Available at: https://europepmc.org/article/med/18457970, Figure 1: https://europepmc.org/articles/PMC2515933/figure/F1/. Figure 3: https://europepmc.org/articles/PMC2515933/figure/F3/. (Accessed: 20 Nov 2022)

(Mastrogiovanni, et al., 2019) Mastrogiovanni, F., Mukhopadhya, A., Lacetera, N., Ryan, M.T., Romani, A., Bernini, R., Sweeney, T., (2019). ‘Anti-Inflammatory Effects of Pomegranate Peel Extracts on In Vitro Human Intestinal Caco-2 Cells and Ex Vivo Porcine Colonic Tissue Explants’. Nutrients. Mar 5;11(3):548. doi: 10.3390/nu11030548. PMID: 30841512; PMCID: PMC6471410. Available at: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6471410/ (Accessed: 22 Nov 2022)

(Milenkovic, et al., 2012) Milenkovic, D., Deval, C., Gouranton, E., Landrier, J.F., Scalbert, A., Morand, C., Mazur, A., (2012). ‘Modulation of miRNA expression by dietary polyphenols in apoE deficient mice: a new mechanism of the action of polyphenols’. PLoS One. 7(1):e29837. doi: 10.1371/journal.pone.0029837. Epub 2012 Jan 10. PMID: 22253797; PMCID: PMC3254631. Available at: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0029837 (Accessed: 27 Nov 2022)

(Monira, et al, 2017) Monira, A.O., Mohammad, M.A., & Ashraf, H.A. (2017). ‘Green biosynthesis of silver nanoparticles using pomegranate peel and inhibitory effects of the nanoparticles on aflatoxin production’. Pakistan Journal of Botany, 49(2), 751-756. Available at: https://inis.iaea.org/search/search.aspx?orig_q=RN:48076454 (Accessed: 7 Nov 2022)

(Nadagouda, et al, 2014) Nadagouda, M., Iyanna, N., Lalley, J., Han, C., Dionysiou, D., Varma, R., (2014). ‘Synthesis of Silver and Gold Nanoparticles Using Antioxidants from Blackberry, Blueberry, Pomegranate, and Turmeric Extracts’. ACS Sustainable Chemistry & Engineering. ACS Publications, Washington, DC, 2(7):1717-1723, (2014). Available at: https://pubs.acs.org/doi/10.1021/sc500237k (Accessed: 16 Nov 2022)

(Nasiriboroumand, et al, 2018)  Nasiriboroumand, M., Montazer, M., Barani, H., 2018. ‘Preparation and characterization of biocompatible silver nanoparticles using pomegranate peel extract’. J Photochem Photobiol B. Feb;179:98-104. doi: 10.1016/j.jphotobiol.2018.01.006. Epub 2018 Jan 10. Available at: https://www.ncbi.nlm.nih.gov/pubmed/29351880 (Accessed: 16 Nov 2022)

(Qabaha, et al., 2019, Figure 1) Qabaha, K., Al-Rimawi, F., Nusseibeh, S., Abbadi, J., Abu-Lafi, S., (2019). ‘Phenolic and Flavonoids Analysis of Pomegranate Peel Extracts and their Anti-inflammatory and Antioxidant Activities’. Int J of Pharmaceutical Quality Assurance 10(1);60-65. ISSN 0975 9506. Available at: https://www.researchgate.net/publication/333651285_Phenolic_and_Flavonoids_Analysis_of_Pomegranate_Peel_Extracts_and_their_Antiniflammatory_and_Antioxidant_Activities  Figure 1: https://www.researchgate.net/figure/Effect-of-pomegranate-peel-extract-on-production-of-IL-6-and-TNF-alpha-by-PMNCs_tbl1_333651285 (Accessed: 22 Nov 2022)

(Rao, et al, 2013) Rao, A., Mahajan, K., Bankar, A., et al., (2013) ‘Facile synthesis of size-tunable gold nanoparticles by pomegranate (Punica granatum) leaf extract: Applications in arsenate sensing’. Applications in arsenate sensing. United States: N. p., 2013. Web. doi:10.1016/J.MATERRESBULL.2012.12.025. Available at:  https://www.osti.gov/etdeweb/biblio/22290393 (Accessed: 16 Nov 2022)

(Saadatmand, et al, 2021) Saadatmand, M., Al-Awsi, G.R.L., Alanazi, A.D., et al., (2021) ‘Green synthesis of zinc nanoparticles using Lavandula angustifolia Vera. Extract by microwave method and its prophylactic effects on Toxoplasma gondii infection’. Saudi J Biol Sci. Nov;28(11):6454-6460. doi: 10.1016/j.sjbs.2021.07.007. Epub 2021 Jul 10. PMID: 34764762; PMCID: PMC8568829. Available at: https://pubmed.ncbi.nlm.nih.gov/34764762/ (Accessed: 16 Nov 2022)

(Sayed, et al, 2022) Sayed, S., Alotaibi, S.S., El-Shehawi, A.M., et al., The Anti-Inflammatory, Anti-Apoptotic, and Antioxidant Effects of a Pomegranate-Peel Extract against Acrylamide-Induced Hepatotoxicity in Rats. Life. 2022; 12(2):224. https://doi.org/10.3390/life12020224 Available at: https://www.mdpi.com/2075-1729/12/2/224/htm (Accessed: 3 November 2022)

(Yang, H., et al, 2016) Yang, H., Ren, Y.-Y., Wang, T., Wang, C., (2016) ‘Preparation and antibacterial activities of Ag/Ag+/Ag3+ nanoparticle composites made by pomegranate (Punica granatum) rind extract’. Results in Physics, 6, pp 299-304. DOI:10.1016/j.rinp.2016.05.012 Available at: http://adsabs.harvard.edu/abs/2016ResPh...6..299Y (Accessed: 21 October 2022)