Headache's Anyone ? (Ammonia, Glutathione, Lysine, Arginine, and High Protein and High Fat Diets!)

For those folks who have long battled a chronic illness, or general feeling of being unwell, headaches and cognitive issues seem par for the course. Sometimes its easy to chalk these up to the usual suspects, who rightfully so can be contributing factors. Things like: mycotoxins (e.g. Ochratoxin A, etc), heavy metal toxicity (e.g. mercury, lead), high levels of dopamine, neuro inflammation, excessive aldehydes from candida, alcohol, excessive glutamate, excessive nitric oxide, oxidative stress from hydroxyl radicals or super oxide, or exposure to toxins like volatile organic compounds. However, observationally, i am seeing some patterns develop genetically that match the symptomology that are unrelated to the above issues and coincide with significant headaches.

Part of being alive is eating protein, and for many of us this means animal protein. Higher animal protein diets have been getting more and more attention in the chronic illness circles for the last few years. Its a lower fiber, and generally lower allergy option to vegetables containing lectins, aquaporins, and various other substances that can cause allergy like reactions in the gut. Its a common approach when dealing with SIBO to cut down on dietary fiber - and animal protein fits the bill. However, with a higher animal protein diet, comes higher ammonia as a by product in the gut. The urea cycle is supposed to take of clearing the ammonia - mostly through the enzymes CPS1, OTC, ASS1, ASL, and ARG1. I inspect over 200 locations on this set of genes for compromise in the Urea Cycle. Significant mutations in these genes are quite rare - both in the genetic database i reference (has 50,000+ clients, mostly chronically unwell), and even more rare against the natural population database found on gnomAD Browser [9], that lists mutation frequency by ethnicity. However, when there are mutations of significance, i have found extra caution around managing ammonia is beneficial. In particular, mutations in ASS1 are super rare, and often screened at birth - with some natural (arginine) and pharmaceutical approaches to reduce urea, nitrogen, and ammonia cited [7]. Depending on the specific mutations, citrulline can be helpful or harmful - to help stimulate the urea cycle.

For those folks who take L-Lysine to manage viral outbreaks or other reasons - it competes with the same transporter as Arginine. Arginine is required for the proper functioning of the Urea Cycle, and neutralizing ammonia. Consistent dosing of L-Lysine is often seen with higher levels of Urea/Ammonia on Comprehensive Metabolic Panels as it can slow down the Urea Cycle when Arginine isnt made available; as is high animal protein diets.

The MAOA (monoamine oxidase A) gene processes amines, and also serves as an escape hatch for excess neurotransmitters. Riboflavin (B2) is the main cofactor, while MAOA exhausts hydrogen peroxide (H₂O₂) and ammonia (NH₃). The amount of hydrogen peroxide generated by MAOA is significant and consumes lots of glutathione. Upregulation mutations of MAOA are consistently seen with lower levels of glutathione on blood panels.

Glutathione is used both to neutralize the hydrogen peroxide [1,2,3], but also ammonia [10]. The specific glutathione genes that are responsible for neutralizing hydrogen peroxide to water are : GPX1, GPX2, GPX3, GPX4, GPX5, GPX6, and GPX7 [1]. See my other article on Antioxidant Systems for information on the GPX enzymes and Catalase as well (primarily expressed in the liver, and also neutralizes hydrogen peroxide). GPX1 is the most ubiquitously expressed GPX enzyme - and is also correlated with all cause mortality, Yikes!. GPX4 is the only GPX enzyme expressed inside the mitochondria, where 75-90% of the body's super oxide is produced [2,3]! SOD2, which is expressed inside the mitochondria, turns Super Oxide into Hydrogen Peroxide, which then gets turned over to GPX4 to process [2,3].

The GPX enzymes are also responsible for neutralizing the hydrogen peroxide produced by processing fatty acids. Significant mutations on GPX1 and or GPX4 spell trouble when combined with mutations in Keap1 and NRF2, both involved in the regulation of anti oxidant systems and the GPX genes [5]. The GSR gene, recycles oxidized glutathione and is critical to maintaining the pool of reduced glutathione, and its primary cofactor is , you guessed it, B2 [5]. Exogenous supplementation of glutathione is best done by liposomal glutathione, but extreme caution should be exercised if mutations in GSR exist - and prior support of GSR should be considered, otherwise the exogenous glutathione may actually increase oxidative stress!

So what does all this mean!? Well, if you have a compromised Urea Cycle, things like a high animal protein diet, a diet high in fatty acids, L-Lysine supplementation, GPX 1 and GPX 4 mutations, Keap1 up regulation and or NRF2 downregulation - you can be setting yourself up for high levels of ammonia in the brain which can lead to headaches and excessive oxidative stress.

What should one do ? Well, we know that Cysteine, Glutamine, and Glycine are the 3 main elements that make up glutathione. Glycine is made in the body from the EAA L-Threonine, by the GCAT enzyme [6]. And you guessed it, folks with lots of mutations on GCAT have lower levels of Glycine and Glutathione, and higher oxidative stress. In a separate article, related to Traumatic Brain Injuries - i will highlight the frequency with which i see GCAT mutations in those who do not heal easily from TBI's. Glycine is also an inhibitory amino acids for neuro transmitters and helps clear excess glutamine, and is found in collagen powders and proteins. NAC (Cysteine) should be considered carefully, if excess unbound iron exists, cysteine will combine with the excess iron and create more oxidative stress through hydroxyl radicals. Glutamine can be healing for the gut lining, but in excess can cause neuro excitatory issues for others.

Things that support the GPX enzyme's are Panax Ginseng. Things that support Keap1 are Sulfurophane. While NRF2 is upregulated by many known substances like Alpha Lipoic Acid, etc. The GSR enzyme is supported by Lycopene and Parsley. A lower protein and lower fat diet, and using fibers like Larch Powder can lower the ammonia in the gut as well. While adding in some arginine, or nuts and chocolate in modest amounts can help stimulate the Urea Cycle [8]. A separate article i will publish on this blog will be related to Nitric Oxide, and how it is stimulated by arginine and nitrates, the good, the bad, and the ugly with NOS1, NOS2, and NOS3 mutations.

[1] First line defence antioxidants-superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX): Their fundamental role in the entire antioxidant defence grid O.M. Ighodaro a,b,⇑ , O.A. Akinloye b. Department of Biochemistry, Faculty of Sciences, Lead City University, Ibadan, Nigeria bDepartment of Biochemistry, College of Biosciences, Federal University of Agriculture, Abeokuta (FUNAAB), Abeokuta, Nigeria. Article history: Received 7 July 2017 Revised 16 August 2017 Accepted 8 September 2017 Available online 13 November 2017.

[2]Mitochondrial Glutathione, a Key Survival Antioxidant Montserrat Marı´,1 Albert Morales,1 Anna Colell,1 Carmen Garcı´a-Ruiz,1 and Jose´ C. Ferna´ ndez-Checa1,2. ANTIOXIDANTS & REDOX SIGNALING Volume 11, Number 11, 2009 ª Mary Ann Liebert, Inc. DOI: 10.1089=ars.2009.2695

[3]Glutathione and mitochondria Vicent Ribas1,2 , Carmen García-Ruiz1,2,3 and José C. Fernández-Checa1,2,3 * 1 Department of Cell Death and Proliferation, Institute of Biomedical Research of Barcelona, Consejo Superior de Investigaciones Científicas (IIBB-CSIC), Barcelona, Spain 2 Liver Unit, Hospital Clínic, Centre Esther Koplowitz, Institut d’Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS)–Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Barcelona, Spain 3 Research Center for Alcoholic Liver and Pancreatic Diseases and Cirrhosis, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA

[4] Mitochondrial Glutathione: Recent Insights and Role in Disease Montserrat Marí 1,* , Estefanía de Gregorio 1 , Cristina de Dios 1,2 , Vicente Roca-Agujetas 1 , Blanca Cucarull 1,2 , Anna Tutusaus 1 , Albert Morales 1,3,* and Anna Colell 1,4,*

[5] Role of Nrf2/HO-1 system in development, oxidative stress response and diseases: an evolutionarily conserved mechanism Agnieszka Loboda1,2 • Milena Damulewicz3 • Elzbieta Pyza3 • Alicja Jozkowicz1 • Jozef Dulak1,2 Received: 27 January 2016 / Revised: 7 April 2016 / Accepted: 8 April 2016 / Published online: 21 April 2016

[6] www.genecards.org. Entrez Gene Summary for GCAT Gene. The degradation of L-threonine to glycine consists of a two-step biochemical pathway involving the enzymes L-threonine dehydrogenase and 2-amino-3-ketobutyrate coenzyme A ligase. L-Threonine is first converted into 2-amino-3-ketobutyrate by L-threonine dehydrogenase. This gene encodes the second enzyme in this pathway, which then catalyzes the reaction between 2-amino-3-ketobutyrate and coenzyme A to form glycine and acetyl-CoA. The encoded enzyme is considered a class II pyridoxal-phosphate-dependent aminotransferase. Alternate splicing results in multiple transcript variants. A pseudogene of this gene is found on chromosome 14. [provided by RefSeq, Jan 2010]

[7] Gene Therapy in Combination with Nitrogen Scavenger Pretreatment Corrects Biochemical and Behavioral Abnormalities of Infant Citrullinemia Type 1 Mice

by Andrea Bazo, Aquilino Lantero, Itsaso Mauleón, Leire Neri, Martin Poms, Johannes Häberle, Ana Ricobaraza, Bernard Bénichou, Jean-Philippe Combal, Gloria Gonzalez-Aseguinolaza, andRafael Aldabe

[8] Clinical Consequences of Urea Cycle Enzyme Deficiencies and Potential Links to Arginine and Nitric Oxide Metabolism. Fernando Scaglia, Nicola Brunetti-Pierri, Soledad Kleppe, Juan Marini, Susan Carter, Peter Garlick, Farook Jahoor, William O'Brien, Brendan Lee. The Journal of Nutrition, Volume 134, Issue 10, October 2004, Pages 2775S–2782S, https://doi.org/10.1093/jn/134.10.2775S Published: 01 October 2004

[9] www.gnnomad.broadinstitute.org