Equol Production Research Articles

Isoflavone biomarkers are inversely associated with atherosclerosis progression in adults: a prospective study

Many studies have examined associations between dietary isoflavones and atherosclerosis, but few used objective biomarkers. We examined the associations of isoflavone biomarkers (primary analyses) and equol production (secondary analyses) with the progression of carotid intima-media thickness (cIMT), and whether inflammation, systolic blood pressure (SBP), blood lipids, and sex hormone-binding globulin (SHBG) mediated these associations, in Chinese adults. This 8.8-y prospective study included 2572 subjects (40-75 y old) from the GNHS (Guangzhou Nutrition and Health Study; 2008-2019). The concentrations of daidzein, genistein, and equol were assayed by an HPLC-tandem MS in serum (n=2572) at baseline and in urine (n=2220) at 3-y intervals. The cIMT of the common carotid artery (CCA) and bifurcation segment were measured by B-mode ultrasound every 3 y, and the progressions of cIMT ( ∆cIMT) were estimated using the regression method. Multivariable linear mixed-effects models (LMEMs) and ANCOVA revealed that subjects with higher serum isoflavones tended to have lower increases of CCA-cIMT. The mean±SEM differences in 8.8-y ∆CCA-cIMT between extreme tertiles of serum isoflavones were -17.1±8.4, -20.6±8.3, and -23.3±10.4 μm for daidzein, total isoflavone, and equol (P-trends<0.05), respectively. LMEMs showed that the estimated yearly changes (95% CIs) (μm/y) in CCA-IMT were -2.0 (-3.8, -0.3), -1.9 (-3.6, -0.1), and -2.1 (-3.8, -0.3) in the highest (compared with the lowest) tertile of daidzein, genistein, and total isoflavones, respectively (P-interaction<0.05). Path analyses indicated that the serum equol-atherosclerosis association was mediated by increased SHBG and decreased SBP. Similar beneficial associations were observed in the secondary analyses. Serum isoflavones and equol exposure were associated with reduced cIMT progression, mediated by SHBG and SBP.

Heterologous expression of equol biosynthesis genes from Adlercreutzia equolifaciens.

ABSTRACTEquol is the isoflavone-derived metabolite with the greatest estrogenic and antioxidant activity. It is produced from daidzein by fastidious and oxygen-susceptible intestinal bacteria, which hinders their use at an industrial scale. Therefore, expressing the equol production machinery into easily-cultivable hosts would expedite the heterologous production of this compound. In this work, four genes (racemase, tdr, ddr and dzr) coding for key enzymes involved in equol production in Adlercreutzia equolifaciens DSM19450T were synthesized and cloned in a pUC-derived vector (pUC57-equol) that was introduced in Escherichia coli. Recombinant clones of E. coli produced equol in cultures supplemented with daidzein (equol precursor) and dihydrodaidzein (intermediate compound). To check whether equol genes were expressed in Gram-positive bacteria, the pUC57-equol construct was cloned into the low-copy-number vector pIL252, and the new construct (pIL252-pUC57-equol) introduced into model strains of Lacticaseibacillus casei and Lactococcus lactis. L. casei clones carrying pIL252-pUC57-equol produced a small amount of equol from dihydrodaidzein but not from daidzein, while L. lactis recombinant clones produced no equol from either of the substrates. This is the first time that A. equolifaciens equol genes have been cloned and expressed in heterologous hosts. E. coli clones harboring pUC57-equol could be used for biotechnological production of equol.

Red clover-rich grassland increases equol concentration in eggs from free-range laying hens

ABSTRACT 1. The aim of this study was to evaluate the production of equol (4ʹ,7-isoflavandiol; a bacterial polyphenol metabolite which is an isoflavandiol oestrogen metabolised from daidzein from plants) enriched eggs from free-range hens fed different pasture species. Four species were tested: red clover, white clover, ryegrass and chicory. 2. The study was conducted from June to September 2017 on eight free range, outdoor areas, each containing fifteen laying hens and sown with a single pasture species 3. Precursors of equol (daidzein, formononetin) were analysed every fortnight from the fresh pasture cover in each area, as well as equol and daidzein levels in eggs. 4. Daidzein and formononetin concentrations in the fresh pasture samples differed significantly according to species (P < 0.001), whereby red clover had the highest concentrations of daidzein and formononetin (85 and 996 µg/g DM, respectively). 5. Equol concentration in eggs differed according to pasture species (P < 0.001). Equol concentrations reached about 1,200 ng/g DM in eggs from hens with access to red clover. These eggs can represent a valuable source of equol in the human diet.

Abstract P137: Association Of Equol Producing Capacity With Aortic Calcification In Middle-aged Japanese Men

Background: Soy isoflavones (ISFs) are regularly consumed in Japan and other Asian countries where some studies showed a significant inverse association of ISFs intake with incident coronary heart disease (CHD). However, a US randomized clinical trial did not find ISFs as anti-atherogenic. This discrepancy may be due to the higher capacity of people in Japan than in the US to produce equol, a metabolite of ISF by the gut microbiome. Equol may have greater anti-atherogenic properties than ISFs. It is unknown whether equol producers (EP), people who produce equol after consuming ISF, have lower aortic calcification (AC), a biomarker of atherosclerosis, as compared to non-EP. Aim: To determine the association between equol producing status and AC in Japanese men. Method: This cross-sectional, population-based study included 302 Japanese men aged 40-49, free of CHD. EP was defined as participants with serum levels of equol ≥20 nM. AC was measured in the entire aorta and quantified by the Agatston method. The presence of AC was defined as AC score ≥10. We analyzed the association between equol producing status and AC using the Tobit and the logistic regressions. Result: We observed 125 participants as EP. In multivariable analyses, EP had non-significantly lower AC score by 147 (95% confidence interval (CI): -386, 92) units and an odds ratio of 0.71 (95% CI: 0.40, 1.26) for the AC presence as compared to non-EP. As significant interaction of equol producing status with age was present, we stratified the analyses by the median age. In men aged 45-49 years, EP had non-significantly lower AC score by 246 units and a non-significant 0.62 times the odds of AC presence as compared to non-EP; the corresponding values in men aged 40-44 were 12 units and 0.91 times (Table). Such an association was not observed with serum ISFs (data not shown). Conclusion: Japanese middle-aged men who were EP had non-significantly lower AC than non-EP and such association was stronger in men aged 45-49 than 40-44 years. Further study with much larger sample size is warranted.

Soy protein supplementation in men following radical prostatectomy: a 2-year randomized, placebo-controlled clinical trial

Many studies have addressed effects of dietary supplementation with soy protein, but most have been inconsistent and few have been long-term studies in men. This study was a secondary analysis of body weight, blood pressure, thyroid hormones, iron status, and clinical chemistry in a 2-y trial of soy protein supplementation in middle-aged to older men. Data were analyzed as secondary outcomes of a randomized controlled trial of dietary supplementation with 20 g/d soy protein isolate, providing 41 mg/d total isoflavones and 23 mg/d genistein, in 44- to 75-y-old men who were at risk of cancer recurrence following prostatectomy randomized to soy (n=50) or a casein-based placebo (n=43). Weight, blood pressure, and blood samples were collected at baseline, every 2 mo in year 1, and every 3 mo in year 2. Compared with casein, soy supplementation did not affect body weight, blood pressure, serum total cholesterol, calcium, phosphorus, and thyroid hormones. Serum ferritin concentrations doubled over 2 y in both groups (117-129%), whereas hemoglobin and hematocrit increased slightly. In an exploratory subgroup analysis of soy group data, weight increased in subjects producing equol but not in nonproducers. Blood pressure was reduced in nonequol producers but not in producers. Other endpoints were not affected by equol production status. Soy protein supplementation for 2 y compared with a casein-based placebo did not affect body weight, blood pressure, serum total cholesterol, iron status parameters, calcium, phosphorus, and thyroid hormones. Exploratory analysis suggests that equol production status of subjects on soy may modify effects of soy on body weight and possibly blood pressure. This trial was registered at clinicaltrials.gov as NCT00765479.

Association of equol with obesity in postmenopausal women.

Equol, which is an active metabolite of isoflavone, has a beneficial impact on metabolic diseases such as dyslipidemia and hyperglycemia. However, the effect of equol on obesity remains uncertain. This study was performed to determine the association between equol and obesity in postmenopausal women. We evaluated 386 women in their 50s-60s who underwent health check-ups from February 2018 to January 2019 at Watari Hospital Health Center in Fukushima, Japan. Overweight and visceral obesity were defined as a body mass index ≥25 kg/m2 and waist circumference ≥ 90 cm, respectively. Participants were asked to complete a questionnaire about their ordinary lifestyle. Participants were defined as equol producers when urinary equol level was 1.0 μM or more. The proportion of individuals with obesity (overweight and visceral obesity) and lifestyle factors were compared between equol producers and nonproducers. In addition, the association between equol and obesity was examined using logistic regression analysis with adjustment for lifestyle factors. Of the 386 participants, 106 (27.5%) women were equol producers. The proportions of women who were overweight (13.2% vs 25.7%) and had visceral obesity (6.6% vs 20.7%) were significantly lower in the equol-producing group than in the nonproducing group. Multivariable logistic regression analysis showed that equol production was significantly associated with overweight (odds ratio =0.47, 95% confidence interval: 0.25-0.88) and visceral obesity (odds ratio =0.30, 95% confidence interval: 0.13-0.68). Equol is significantly associated with obesity in postmenopausal women.

A Systematic Review of the Effects of Equol (Soy Metabolite) on Breast Cancer.

Equol is a soy isoflavone metabolite that can be produced by intestinal bacteria. It is lipophilic and resembles natural oestrogens with an affinity to oestrogen receptors. This review is focused on how equol affects breast cancer, as evidenced by in vivo and in vitro studies. Equol is considered chemoprotective in specific endocrine-related pathologies, such as breast cancer, prostate cancer, cardiovascular diseases, and menopausal symptoms. In humans, not everyone can produce equol from gut metabolism. It is postulated that equol producers benefit more than non-equol producers for all the endocrine-related effects. Equol exists in two enantiomers of R-equol and S-equol. Earlier studies, however, did not specify which enantiomer was being used. This review considers equol’s type and concentration variations, pathways affected, and its outcome in in vivo and in vitro studies.

Administration of Cholic Acid Inhibits Equol Production from Daidzein in Mice.

Equol (Eq) is a metabolite of soy isoflavone daidzein (De) produced by the intestinal microbiota. The clinical effectiveness of soy isoflavone is considered to depend on the individual ability of Eq production. Previous studies have demonstrated that habitual dietary patterns may influence the production of Eq. For example, high Eq producers consumed less fat as a percentage of energy than low Eq producers. However, the inhibitory factors of Eq production are unknown. Recently, it was reported that bile acids induced by high-fat diet consumption may be a host-related factor controlling the composition of the intestinal microbiota. In this study, we investigated the effect of cholic acid (CA) administration, a mimic of the microbiota altered by a high-fat diet, on Eq production in mice. CA administration significantly decreased the levels of the De metabolites Eq, dihydrodaidzein, and O-desmethylangolensin in the serum of mice. However, CA administration did not affect the total molar concentration of De and its metabolites. Moreover, CA administration increased the levels of secondary bile acids, particularly deoxycholic acid (DCA), which has strong antibacterial activity in the cecum contents of mice. Thus, CA administration may increase the levels of DCA, a secondary bile acid, resulting in inhibition of Eq production. These findings may help to reveal the factors inhibiting Eq production and enhance the clinical effectiveness of isoflavone intake.

Equol-producing status affects exercise training-induced improvement in arterial compliance in postmenopausal women.

Central arterial compliance decreases drastically after menopause. Regular intake of soy isoflavone and aerobic exercise increase arterial compliance. The equol is a metabolite of isoflavone daidzein by gut microbiome. We determined whether the equol-producing status affects aerobic exercise-induced improvement in carotid arterial compliance. Forty-three postmenopausal women were assigned to two intervention groups: 1) exercise and isoflavone (Ex+Iso, n = 27 females) or 2) isoflavone interventions (Iso; n = 16 females). Participants of the Ex+Iso intervention group completed an 8-wk aerobic exercise training, and all participants were administered with oral isoflavone supplements during the interventions. The equol-producing status (equol producers or nonproducers) was determined from urine equol concentrations after a soy challenge. In the Ex+Iso intervention group, carotid arterial compliance increased in the equol producers (0.084 ± 0.030→0.117 ± 0.035 mm2/mmHg), but not in the nonproducers (0.089 ± 0.028→0.097 ± 0.026 mm2/mmHg) after the intervention (interaction effect; P < 0.05). The magnitude of increases in carotid arterial compliance was significantly greater in the equol producers than in the non-equol producers (P < 0.05). In the isoflavone intervention group, there were no changes in any parameters after the intervention irrespective of the equol status. These results suggest that equol-producing status is obligatory to aerobic exercise training-induced improvements in central arterial compliance in postmenopausal women.NEW & NOTEWORTHY Isoflavone intake and aerobic exercise increase central artery compliance. Equol, a metabolite of isoflavone daidzein by gut microbiome, has beneficial effects on vascular function. We demonstrated for the first time that the interaction of aerobic exercise and equol production status plays an essential role in improvements in central artery compliance in postmenopausal women. More specifically, the equol-producing status was obligatory to exercise training-induced improvements in central arterial compliance in postmenopausal women.

Impact of prebiotics on equol production from soymilk isoflavones by two Bifidobacterium species

The influence of commercial prebiotics (fructo-oligosaccharides and inulin) and sugars (glucose and sucrose) on enhancing equol production from soymilk isoflavones by Bifidobacterium longum BB536 and Bifidobacterium breve ATCC 15700 was evaluated in vitro. Sterilized soymilk was inoculated with each bacterial species at 37 °C for 48 h. The growth and β-glucosidase enzyme activity for the two Bifidobacterium species in soymilk throughout fermentation were assessed. The highest viable count for B. breve (8.75 log CFU/ml) was reached at 36 h and for B. longum (8.55 log CFU/ml) at 24 h. Both bacterial species displayed β-glucosidase activity. B. breve showed increased enzyme activity (4.126 U) at 36 h, while B. longum exhibited maximum activity (3.935 U) at 24 h of fermentation. Among the prebiotics screened for their effect in isoflavones transformation to equol, inulin delivered the highest effect on equol production. The co-culture of B. longum BB536 and B. breve ATCC15700 in soymilk supplemented with inulin produced the highest level (11.49 mmol/l) of equol at 48 h of fermentation process. Level of daidzin declined whereas that of daidzein increased, and then gradually decreased due to formation of equol when soymilk was fermented using bifidobacterial. This suggests that the nutritional value of soymilk may be increased by increasing bioavailability of the bioactive ingredients. Collectively these data identify probiotics and prebiotic combinations suitable for inclusion in soymilk to enhance equol production.

Metabolism of Soy Isoflavones by Intestinal Bacteria: Genome Analysis of an Adlercreutzia Equolifaciens Strain That Does Not Produce Equol.

Isoflavones are transformed in the gut into more estrogen-like compounds or into inactive molecules. However, neither the intestinal microbes nor the pathways leading to the synthesis of isoflavone-derived metabolites are fully known. In the present work, 73 fecal isolates from three women with an equol-producing phenotype were considered to harbor equol-related genes by qPCR. After typing, 57 different strains of different taxa were tested for their ability to act on the isoflavones daidzein and genistein. Strains producing small to moderate amounts of dihydrodaidzein and/or O-desmethylangolensin (O-DMA) from daidzein and dihydrogenistein from genistein were recorded. However, either alone or in several strain combinations, equol producers were not found, even though one of the strains, W18.34a (also known as IPLA37004), was identified as Adlercreutzia equolifaciens, a well-described equol-producing species. Analysis and comparison of A. equolifaciens W18.34a and A. equolifaciens DSM19450T (an equol producer bacterium) genome sequences suggested a deletion in the former involving a large part of the equol operon. Furthermore, genome comparison of A. equolifaciens and Asaccharobacter celatus (other equol-producing species) strains from databases indicated many of these also showed deletions within the equol operon. The present results contribute to our knowledge to the activity of gut bacteria on soy isoflavones.

In vitro metabolism of red clover isoflavones in rumen fluid.

The degradation of red clover isoflavones was studied in vitro using a rumen fluid buffer system. Various amounts of red clover extract (5-75mg) together with hay or concentrate-rich diet were added to 40ml of rumen fluid obtained from non-lactating and lactating dairy cows, respectively, and incubated for 0, 3, 6, 12 or 24hr. Following incubation, concentrations of daidzein, genistein, formononetin, biochanin A and equol were determined in the samples. After 3hr of incubation, isoflavone metabolism and equol production could be observed. The results obtained indicate that hay diet provides better conditions for isoflavone metabolism, as concentrations of daidzein, formononetin and biochanin A were higher in incubations based on the concentrate-rich diet and the production of equol was higher in incubations based on the hay diet. Furthermore, in incubations with higher amounts of added clover extract, a decrease in equol production was observed. Further studies are needed to clarify the role of adaptation of rumen microflora on isoflavone degradation kinetics and to clarify the interrelationship between various dietary factors, rumen microbiota and isoflavones. The knowledge of isoflavone metabolism kinetics in dependence on studied factors will be useful for the optimization of feeding dose.

Colonization Potential to Reconstitute a Microbe Community in Pseudo Germ-Free Mice After Fecal Microbe Transplant From Equol Producer.

Human intestinal microbiota plays a crucial role in the conversion of isoflavones into equol. Usually, human microbiota-associated (HMA) animal models are used, since it is difficult to establish the mechanism and causal relationship between equol and microbiota in human studies. Currently, several groups have successfully established HMA animal models that produce equol through germ-free mice or rats; however, the HMA model of producing equol through pseudo germ-free mice has not been established. The objective of this study is to establish an HMA mice model for equol production through pseudo germ-free mice, mimicking the gut microbiota of an adult human equol producer. First, a higher female equol producer was screened as a donor from 15 volunteers. Then, mice were exposed to vancomycin, neomycin sulfate, metronidazole, and ampicillin for 3 weeks to obtain pseudo germ-free mice. Finally, pseudo germ-free mice were inoculated with fecal microbiota of the equol producer for 3 weeks to establish HMA mice of producing equol. The results showed that (i) the ability to produce equol was partially transferred from the donor to the HMA mice. (ii) Most of the original intestinal microbiota of mice were eliminated after broad-spectrum antibiotic administration. (iii) The taxonomy data from HMA mice revealed similar taxa to the donor sample, and the species richness returned to the level close to the donor. (iv) The family Coriobacteriaceae and genera Collinsella were successfully transferred from the donor to HMA mice. In conclusion, the HMA mice model for equol production, based on pseudo germ-free mice, can replace the model established by germ-free mice. The model also provides a basis for studying microbiota during the conversion from isoflavones into equol.

Impact of Short-Term Isoflavone Intervention in Polycystic Ovary Syndrome (PCOS) Patients on Microbiota Composition and Metagenomics.

Background: Polycystic ovary syndrome (PCOS) affects 5–20% of women of reproductive age worldwide and is associated with disorders of glucose metabolism. Hormone and metabolic signaling may be influenced by phytoestrogens, such as isoflavones. Their endocrine effects may modify symptom penetrance in PCOS. Equol is one of the most active isoflavone metabolites, produced by intestinal bacteria, and acts as a selective estrogen receptor modulator. Method: In this interventional study of clinical and biochemical characterization, urine isoflavone levels were measured in PCOS and control women before and three days after a defined isoflavone intervention via soy milk. In this interventional study, bacterial equol production was evaluated using the log(equol: daidzein ratio) and microbiome, metabolic, and predicted metagenome analyses were performed. Results: After isoflavone intervention, predicted stool metagenomic pathways, microbial alpha diversity, and glucose homeostasis in PCOS improved resembling the profile of the control group at baseline. In the whole cohort, larger equol production was associated with lower androgen as well as fertility markers. Conclusion: The dynamics in our metabolic, microbiome, and predicted metagenomic profiles underline the importance of external phytohormones on PCOS characteristics and a potential therapeutic approach or prebiotic in the future.

Application of recombinant lactic acid bacteria and bifidobacteria able to enrich soy beverage in dihydrodaidzein and dihydrogenistein

Dihydrodaidzein (DHD) and dihydrogenistein (DHG) are intermediate compounds in the production of equol and 5-hydroxy-equol from daidzein and genistein by certain intestinal bacteria. In this work, we explored the heterologous expression of the daidzein reductase gene from Slackia isoflavoniconvertens DSM22006, responsible for the formation of DHD and DHG, in nine lactic acid bacteria and Bifidobacterium strains under a strong constitutive promoter in the plasmid pNZ:TuR.dzr. All the transformed strains showed high production of DHD and DHG both from pure daidzein and genistein and from the isoflavone glycosides present in soy beverage. In addition, Lactococcus lactis MG1363 pNZ:TuR.dzr, expressing the recombinant daidzein reductase, incremented the production of equol by equol-producing intestinal microbiotas in a colonic environment. Nevertheless, other recombinant strains tested showed the opposite effect to L. lactis MG1363 pNZ:TuR.dzr in the production of equol by intestinal microbiota. Here, we describe for the first time the production of 5-hydroxy-equol by human intestinal microbiota, both from genistein and DHG. The use of DHD and DHG as substrates, compared to daidzein and genistein, resulted in an increment of equol and 5-hydroxy-equol production by intestinal microbiota. The recombinant strains developed would be of value for the development of fermented soy beverage enriched in DHD and DHG with the aim of facilitate equol and 5-hydroxy-equol production by intestinal microbiota.

Maturation of Vaginal Epithelium and Dyspareunia Symptoms in Equol Producing and Non-Producing Menopausal Women

Introduction: Equol is a metabolite of soy isoflavon called daidzein which is produced by gastrointestinal tract bacteria. This research aims to analyze the maturation of vaginal epithelium and dyspareunia symptoms in producing and non-producing equol menopausal women.Method: This is a cross sectional research. Subject was a community of menopausal women who fulfilled inclusion criteria. Subjects were asked to sign a written informed consent. Subjects underwent vaginal epithelium maturation assessment and were asked whether she experienced dyspareunia. Research was conducted in January 2017.Result: There was a significant difference on the maturation of vaginal epithelium and dyspareunia symptoms between equol producing and non-producing women (p &lt; 0.05). This research found that in equol producing menopausal women, there was a shift-to-the-right vaginal epithelium maturation with more superficial cells compared to parabasal cells produced and less dyspareunia. Meanwhile, in women who did not produce equol, there was a shift-to-the-left vaginal epithelium maturation with more parabasal cells compared to superficial cells produced and more dyspareunia.Conclusion: In equol producing menopausal women, vaginal epithelium will undergo a shift-to-the-right maturation, with more superficial cells produced compared to women who did not produce equol.Maturasi Epitel Vagina dan Gejala Dispareunia pada Wanita Menopause yang Menghasilkan Equol dan Wanita yang Tidak Menghasilkan EquolAbstrakPendahuluan: Equol adalah metabolit isoflavon kedelai yang disebut daidzein yang diproduksi oleh bakteri saluran pencernaan. Penelitian ini bertujuan untuk menganalisis maturasi epitel vagina dan gejala dispareunia pada wanita menopause yang memproduksi dan tidak memproduksi equol.Metode: Penelitian ini merupakan penelitian cross sectional. Subjek penelitian adalah sekelompok wanita menopause yang memenuhi kriteria inklusi. Subjek diminta untuk menandatangani persetujuan tertulis dan menjalani penilaian maturasi epitel vagina dan ditanya apakah mengalami dispareunia. Penelitian dilakukan pada Januari 2017.Hasil: Terdapat perbedaan yang signifikan pada maturasi epitel vagina dan gejala dispareunia antara wanita yang memproduksi equol dan yang tidak memproduksi (p &lt;0,05). Pada wanita menopause yang memproduksi equol, terjadi pematangan epitel vagina shift-to-the-right disertai produksi sel superfisial yang lebih banyak dibandingkan sel parabasal dan lebih sedikit gejala dispareunia. Sementara itu, pada wanita yang tidak menghasilkan equol, terjadi pergeseran shift-to-the-left maturasi epitel vagina dengan produksi sel parabasal lebih banyak dibandingkan sel superfisial dan lebih sering gejala dispareunia.Kesimpulan: Wanita menopause yang memproduksi equol mengalami maturasi epitel vagina shift-to-the-right disertai produksi sel superfisial yang lebih banyak dibandingkan dengan wanita menopause yang tidak memproduksi equol.Kata kunci: Dyspareunia; equol; maturasi epitel vagina

Maturation of Vaginal Epithelium and Dyspareunia Symptoms in Equol Producing and Non-Producing Menopausal Women

Introduction : Equol is a metabolite of soy isoflavon called daidzein which is produced by gastrointestinal tract bacteria. This research aims to analyze the maturation of vaginal epithelium and dyspareunia symptoms in producing and non-producing equol menopausal women. Method : This is a cross sectional research. Subject was a community of menopausal women who fulfilled inclusion criteria. Subjects were asked to sign a written informed consent. Subjects underwent vaginal epithelium maturation assessment and were asked whether she experienced dyspareunia. Research was conducted in January 2017. Result : There was a significant difference on the maturation of vaginal epithelium and dyspareunia symptoms between equol producing and non-producing women (p &lt; 0.05). This research found that in equol producing menopausal women, there was a shift-to-the-right vaginal epithelium maturation with more superficial cells compared to parabasal cells produced and less dyspareunia. Meanwhile, in women who did not produce equol, there was a shift-to-the-left vaginal epithelium maturation with more parabasal cells compared to superficial cells produced and more dyspareunia. Conclusion : In equol producing menopausal women, vaginal epithelium will undergo a shift-to-the-right maturation, with more superficial cells produced compared to women who did not produce equol. Maturasi Epitel Vagina dan Gejala Dispareunia pada Wanita Menopause yang Menghasilkan Equol dan Wanita yang Tidak Menghasilkan Equol Abstrak Pendahuluan : Equol adalah metabolit isoflavon kedelai yang disebut daidzein yang diproduksi oleh bakteri saluran pencernaan. Penelitian ini bertujuan untuk menganalisis maturasi epitel vagina dan gejala dispareunia pada wanita menopause yang memproduksi dan tidak memproduksi equol. Metode : Penelitian ini merupakan penelitian cross sectional. Subjek penelitian adalah sekelompok wanita menopause yang memenuhi kriteria inklusi. Subjek diminta untuk menandatangani persetujuan tertulis dan menjalani penilaian maturasi epitel vagina dan ditanya apakah mengalami dispareunia. Penelitian dilakukan pada Januari 2017. Hasil : Terdapat perbedaan yang signifikan pada maturasi epitel vagina dan gejala dispareunia antara wanita yang memproduksi equol dan yang tidak memproduksi (p &lt;0,05). Pada wanita menopause yang memproduksi equol, terjadi pematangan epitel vagina shift-to-the-right disertai produksi sel superfisial yang lebih banyak dibandingkan sel parabasal dan lebih sedikit gejala dispareunia. Sementara itu, pada wanita yang tidak menghasilkan equol, terjadi pergeseran shift-to-the-left maturasi epitel vagina dengan produksi sel parabasal lebih banyak dibandingkan sel superfisial dan lebih sering gejala dispareunia. Kesimpulan: Wanita menopause yang memproduksi equol mengalami maturasi epitel vagina shift-to-the-right disertai produksi sel superfisial yang lebih banyak dibandingkan dengan wanita menopause yang tidak memproduksi equol. Kata kunci : Dyspareunia; equol; maturasi epitel vagina

Soy Metabolism by Gut Microbiota from Patients with Precancerous Intestinal Lesions.

Background: Colorectal cancer (CRC) requires the presence of a variety of factors predisposing a tumorigenic milieu. Excluding familial clustering and hereditary CRC syndromes, the development of sporadic CRC from precancerous lesions is influenced by tissue inflammation, modulation of intestinal immunity, hormones, dietary habits and gut microbiota composition. As concerning the last two aspects, the intestinal presence of equol, the most biologically active metabolite of the soy isoflavone daidzein and the presence of a genetic determinant of gut microbiota able to metabolize daidzein, seem to lower the CRC risk. It has been hypothesized that the anaerobic microorganisms of the Bacteroides genus play a role in equol production. Aim: To evaluate the presence of (i) anaerobic gut microbiota and (ii) the urinary levels of soy isoflavones (daidzein, genistein and equol) in patients with and without precancerous lesions, challenged with a daidzein-rich soy extract. Methods: Consecutive subjects undergoing colonoscopy participated to the study. Feces were collected from all patients one week before colonoscopy for gut microbiota studies. After the endoscopy examination and the histological evaluation, 40 subjects, 20 with sporadic colorectal adenomas (SCA/P group) and 20 without proliferative lesions (control group) were enrolled for the study. Urine levels of soy isoflavones daidzein, genistein and their metabolite equol, were determined by high performance liquid chromatographic (HPLC) analysis and gut microbiota analysis was performed by Matrix Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS) procedure. Results: Seventeen different bacterial species were identified in the fecal samples of the forty subjects participating to the study. Ten bacterial species resulted anaerobic Gram-negative bacteria, all belonging to the Bacteroides genus. A significant difference of bacteria species was evidenced in the fecal samples of the two groups of subjects. Particularly important was the evidence of Parabacteroides distasonis, Clostridium clostridioforme and Pediococcus pentasaceus only in control fecal samples, such as the presence of Bacteroides fragilis and Prevotella melaningenica only in SCA/P fecal samples. Concerning the soy isoflavones levels, no statistically significant differences were revealed in the genistein and daidzein urinary levels between the two groups of subjects. On the contrary, urinary equol levels were undetectable in ten SCA/P subjects and in two controls; moreover, when present, the levels of urinary equol were significantly lower in SCA/P subjects compared to controls (0.24 ± 0.27 mg/24 hrs vs. 21.25 ± 4.3 mg/24 hrs, respectively, p = 1.12 × 10−6). Conclusions: Our results suggest that the presence of anaerobic Bacteroides in the colon, and the production of equol from soy, could determine a milieu able to contrast the development of colonic mucosa proliferative lesions.

Draft Genome Sequence of Adlercreutzia equolifaciens IPLA 37004, a Human Intestinal Strain That Does Not Produce Equol from Daidzein.

Equol is an intestinal bacterial metabolite derived from the isoflavone daidzein and has beneficial health effects. Most equol producers belong to the family Coriobacteriaceae, which includes species such as Adlercreutzia equolifaciens Here, we report the draft genome sequence of A. equolifaciens IPLA 37004, a human isolate that does not produce equol.

Modulation of equol production via different dietary regimens in an artificial model of the human colon

• This search aimed to find dietary conditions enhancing endogenous equol production. • Control and experimental diets with isoflavones were run in an artificial intestine. • The model was inoculated with a pooled faecal homogenate from equol-producing women. • A carbohydrate-rich diet was found to favour the synthesis of equol. In order to find dietary conditions favouring endogenous equol biosynthesis, a pooled faecal homogenate from equol-producing women was used to inoculate the TIM-2 artificial model of the human proximal colon. The model was fuelled with control diets not supplemented (C) or supplemented (C-ISO) with isoflavones, and two isoflavone-containing diets rich in carbohydrate (CH-ISO) or protein (PR-ISO). Compared to the C-ISO control, the CH-ISO diet doubled the production of equol, while with the PR-ISO diet the production of equol in cultures decreased sharply. The CH-ISO diet was also associated with enhanced butyrate production. The numbers of most bacterial populations analysed did not significantly change along cultures with any of the diets. Surprisingly, counts for a gene involved in equol production ( tdr ) were reduced in all cultures, reflecting a reduction in the number of equol-producing bacteria . In conclusion, under the TIM-2 culture conditions established, the CH-ISO diet favoured the synthesis of equol.