4-OHE1and 4-OH-E2 are referred to as the “bad” estrogens, along with 16a-OHE1. They are primarily produced by CYP1B1 and CYP34A, respectively, enzymes localized in tissues, including breast and prostate as well as liver. Some have suggested that increased expression of CYP1B1 and 4-hydroxylation of estradiol are biomarkers of tumorigenesis.

Human breast cancer tissue produces much higher levels of 4-OH than 2-OH, while normal breast tissue produces approximately equal amounts of the two metabolites. Women taking hormone therapy with a polymorphism in CYP1B1 had twice the risk of developing breast cancer compared to other HRT users.

The hydroxylation of estradiol is one of the major routes of metabolism of the estrogen steroid hormone estradiol. It is hydroxylated into the catechol estrogens 2-hydroxyestradiol and 4-hydroxyestradiol and into estriol (16α-hydroxyestradiol). 4-hydroxyestradiol (4-OH-E2), like 2-OH-E2, can be physiologically active as well as tumorigenic. 4-OH-E2 is capable of binding estrogen receptors with a reduced dissociation rate and prolonged activation, thereby inducing cellular growth and proliferation, adenohypophyseal hormone secretion, and prostaglandin production.

- The 4-OH pathway is considered the most genotoxic as its metabolites can create reactive products that damage DNA.

- Estrone is hydroxylated through Phase 1 detox to form 4-OH-E1. In Phase 2 it is methylation to form 4-MeE1. When 4-OH-E1 is properly methylated to 4-MeE1 it is relatively benign as the 4-MeE1 is easily eliminated and risks are low.

- When it is not methylated 4-OH-E1 builds up. Then it converts to 3,4-Quinones which are carcinogenic similarly to the 16 pathway. Women with uterine fibroids may have increased levels of 4-OH-E1. High levels of estrogen across the board are associated with heavy cycles.

- The 4-OH pathway is considered the most genotoxic as its metabolites can create reactive products that damage DNA.

- Estrone is hydroxylated through Phase 1 detox to form 4-OH-E1. In Phase 2 it is methylation to form 4-MeE1. When 4-OH-E1 is properly methylated to 4-MeE1 it is relatively benign as the 4-MeE1 is easily eliminated and risks are low.

- When it is not methylated 4-OH-E1 builds up. Then it converts to 3,4-Quinones which are carcinogenic similarly to the 16 pathway. Women with uterine fibroids may have increased levels of 4-OH-E1. High levels of estrogen across the board are associated with heavy cycles.

Percentages of 2-OH-E1, 4-OH-E1, and 16-OH-E1

When evaluating phase I metabolism, it can be helpful to compare the percentages of 2, 4, and 16 OH-E1 metabolites. Most individuals metabolize the majority of their estrogens down the 2-OH-E1 pathway which is generally considered the “safer pathway”. This is followed by 16-OH-E1 and 4-OH-E1 respectively, both of which are deemed more reactive and potentially genotoxic.

In a Hormone Metabolite Assessment Panel (HUMAP) designed for pre-menopausal women, the percentage of 4-Hydroxyestrone (4-OH-E1) plays a crucial role in understanding estrogen metabolism and its associated risks. 4-OH-E1 is one of the metabolites of estrogen produced through the 4-hydroxylation pathway, which is significant due to its potent biological activity. This metabolite is known for its strong estrogenic effects and has been linked to an increased risk of estrogen-related cancers, particularly breast cancer.

In the body, 4-OH-E1 can be further metabolized to form catechol estrogen quinones, which have the potential to damage DNA and cause mutations. The percentage of 4-OH-E1 in the overall estrogen metabolite profile is a critical marker; higher percentages can indicate a greater reliance on the 4-hydroxylation pathway, suggesting a potential increase in the risk for estrogen-driven conditions. Factors influencing the proportion of 4-OH-E1 include genetics, liver health, diet, lifestyle, and exposure to environmental toxins.

A very carcinogenic estrogen metabolite, levels low in the reference range are desirable. Additional magnesium, liver support, and methylation support may help decrease 4-OH-E1 levels.

- The 4-OH pathway is considered the most genotoxic as its metabolites can create reactive products that damage DNA.

- Estrone is hydroxylated through Phase 1 detox to form 4-OH-E1. In Phase 2 it is methylation to form 4-MeE1. When 4-OH-E1 is properly methylated to 4-MeE1 it is relatively benign as the 4-MeE1 is easily eliminated and risks are low.

- When it is not methylated 4-OH-E1 builds up. Then it converts to 3,4-Quinones which are carcinogenic similarly to the 16 pathway. Women with uterine fibroids may have increased levels of 4-OH-E1. High levels of estrogen across the board are associated with heavy cycles.

4 hydroxy estrone (4-OH-E1) and estradiol (4-OH-E2) are metabolites of estrone and estradiol and are very reactive estrogens. They are highly prone to the formation of catechol estrogen-derived 3,4 semi-quinones, which are potent, electrophilic, free radical-generating molecules that have been shown to lead to DNA mutagenesis. Indeed, 4 hydroxy E1 and E2 are the most potent and potentially carcinogenic estrogens.

4 hydroxy estrone (4-OH-E1) and estradiol (4-OH-E2) are metabolites of estrone and estradiol and are very reactive estrogens. They are highly prone to the formation of catechol estrogen-derived 3,4 semi-quinones, which are potent, electrophilic, free radical-generating molecules that have been shown to lead to DNA mutagenesis. Indeed, 4 hydroxy E1 and E2 are the most potent and potentially carcinogenic estrogens.

The 41 KD band is often found on the Western blot. A recent study looked at the banding patterns of patients with chronic Lyme disease and healthy controls from the inner city of New York who have never had Lyme disease. They found that a large percentage of the healthy controls tested positive on the 41kd band.

Two types of antibodies are detected in the Western blot test.

This particular marker is called 41 KD (IGM) and hence is a IgM antibody marker. IgM antibodies reflect a relatively recent infection. IgG antibodies in contrast are a sign of an older infection.

Two types of antibodies are detected in the Western blot test.

This particular marker is called 45 KD (IGG) Band and hence is a IgG antibody marker. IgG antibodies are a sign of an older infection. In contrast, IgM antibodies reflect a relatively recent infection. 

IgM antibodies usually disappear after eight weeks post-exposure. 
IgG remains in the serum for a very long time. 

Phenylacetic acid (PAA) and phenylpropionic acid (PPA) are products of phenylalanine metabolism caused by intestinal bacteria. High levels of PAA or PPA in urine may result from the dysbiosis of intestinal flora or the decreased metabolism of phenylalanine, such as phenylketonuria (PKU). PAA is also a metabolite of 2-phenylethylamine, and low levels of PAA in urine are considered as a marker of depression. Urinary 4-hydroxybenzoic acid (4-HBA) and 4-hydroxyphenylacetic acid (4-HPAA) represent a considerable percentage of tyrosine intake. 4-HBA is one of the major catechin metabolites after the intake of green tea infusions. In addition, 4-HPAA has been found to be useful in screening for small bowel diseases associated with anaerobic bacterial overgrowth.

5 Alpha-Dihydrotestosterone (DHT) is a hormone that plays a crucial role in male development and health. It is derived from testosterone, the primary male sex hormone, through the action of an enzyme called 5-alpha-reductase. DHT is significantly more potent than testosterone and is involved in a variety of physiological processes. During puberty, DHT is responsible for the development of secondary sexual characteristics in males, such as facial and body hair, deepening of the voice, and growth of the genitals. It also plays a role in the growth and maintenance of the prostate gland. However, DHT is also associated with male pattern baldness and can contribute to prostate enlargement and other health issues later in life.