Phytoestrogen Content Research Articles

Sex-specific associations between dietary legume subtypes and type 2 diabetes in a prospective cohort study.

Dietary soy, known for its high phytoestrogen content, has been suggested to exhibit a sex-specific association with type 2 diabetes. However, evidence regarding the sex-specific associations of different legume subtypes with type 2 diabetes remains scarce. We aimed to evaluate whether habitual consumption of soy and non-soy legumes (beans and peanuts) was prospectively and sex-specifically associated with the risk of type 2 diabetes incidence, taking into considering significant sex-specific genetic factors beyond legume consumption. A total of 16,666 participants (96,945 person-years) were followed and 945 incident cases were observed. Cumulative intake of legume subtypes was calculated using a food frequency questionnaire administered at baseline and during the revisit surveys. Non-soy legumes are inversely associated with type 2 diabetes in both men and women. Dietary soy intake, however, demonstrated a unilaterally interacting sex-specific association with type 2 diabetes risk (pinteraction for sex=0.0173). Specifically, there was a significant inverse association with type 2 diabetes risk in women (incidence rate ratio, 0.66; 95% confidence interval, 0.48 to 0.80; ptrend=0.0066), but no such association was observed in men. This sex-specific association persisted and even appeared antagonistic in minor allele carriers of 2 novel single nucleotide polymorphisms, rs10196939 (LRRTM4) and rs11750158 (near GFPT2) (pinteraction for sex=0.0014 and 0.0112, respectively). Habitual consumption of legumes shows protective impacts against type 2 diabetes, although these benefits vary by sex. Non-soy legumes provide health advantages for both men and women, whereas soy consumption seems to be beneficial exclusively for women.

COMPARISON IN DNA BARCODING, GENE EXPRESSION AND CHEMICAL OF PUERARIA MIRIFICA PLANTS

The Pueraria mirifica plant is used as a precious material in healthcare and herbal supplement production as it is rich in phytoestrogen content. The wide cultivation gradually directs to the differences among cultivars based on their local original planting region. Besides variation in morphology, P. mirifica plants are different in phytoestrogen content. With barcoding markers located on nuclear and chloroplast, our phylogenetic analysis proved that using the group of matK, rpoC, rps16, and trnH-psbA concatenated sequences provides better discriminating power than using a single marker of common P. mirifica cultivars. Only the maximum likelihood tree based on marker combination separate P. mirifica plants from other members of the Pueraria family and separated tribes in the Fabaceae family. In addition, daidzein, genistin, and puerarin contents were recorded from the tuber of all cultivars. These results showed that some are potential cultivars for extracting phytoestrogens from the tuber. Among the five varieties studied, NA has the highest content of Daidzein and Puerarin, and TLBYT has the most difference in the sequences of rps16 and trnH-psbA and the lowest content of Puerarin, Genistin, and Daidzein. Together, qRT-PCR was performed to evaluate eight phytoestrogen biosynthesis gene expressions in leaf and tuber, in which four novel genes CHS11, CHS13, CHI2A, and UGT2 were examined for the first time in P. mirifica. These gene expressions varied depending on organs and cultivars. The differences in both the content of substances and in DNA barcodes as well as gene expression showed a clear polymorphism between varieties. The information is valuable for the selection of material sources for production. Keywords: DNA barcoding, P. mirifica, CHS, CHI, UGT.

Temporal analysis of water chemistry and smallmouth bass (Micropterus dolomieu) health at two sites with divergent land use in the Susquehanna River watershed, Pennsylvania, USA

Monitoring wild fish health and exposure effects in impacted rivers and streams with differing land use has become a valuable research tool. Smallmouth bass (Micropterus dolomieu) are a sensitive, indicator species that exhibit signs of immunosuppression and endocrine disruption in response to water quality changes and contaminant exposure. To determine the impact of agriculture and development on smallmouth bass health, two sites (a developed/agriculture site and a forested site) in the Susquehanna River watershed, Pennsylvania were selected where bass and water chemistry were sampled from 2015 to 2019. Smallmouth bass were sampled for histopathology to assess parasite and macrophage aggregate density in the liver and spleen, condition factor (Ktl), hepatic gene transcript abundance, hepatosomatic index (HSI), and a health assessment index (HAI). Land use at the developed/agriculture site included greater pesticide application rates and phytoestrogen crop cover and more detections and higher concentrations of pesticides, wastewater-associated contaminants, hormones, phytoestrogens, and mycotoxins than at the forested site. Additionally, at the developed/agriculture site, indicators of molecular changes, including oxidative stress, immune/inflammation, and lipid metabolism-related hepatic gene transcripts, were associated with more contaminants and land use variables. At both sites, there were multiple associations of contaminants with liver and/or spleen macrophage aggregate density, indicating that changes at the molecular level seemed to be a better indicator of exposures unique to each site. The findings illustrate the importance of timing for land management practices, the complex mixtures aquatic animals are exposed to, and the temporal changes in contaminant concentration. Agricultural practices that affect hepatic gene transcripts associated with immune function and disease resistance were demonstrated which could negatively affect smallmouth bass populations.

Comparative Analysis of Grape Seed Oil, Linseed Oil, and a Blend: In Vivo Effects of Supplementation.

Grape seeds are rich in bioactive substances, including polyphenols, terpenoids, and phytosterols. Linseed (Linum usitatissimum L.) boasts a high concentration of polyunsaturated fatty acids (PUFAs), lignans, phytoestrogens, and soluble fibers, all contributing to its therapeutic potential. In this study, we pioneered the formulation of an oil blend (GL) combining grape seed oil (G) and golden linseed oil (GL) in equal volumes (1:1 (v/v)) and we evaluated in terms of the nutritional, physical, and chemical properties and their influence in an in vivo experimental model. We analyzed the oils by performing physical-chemical analyses, examining the oxidative stability using Rancimat; conducting thermal analyses via thermogravimetry/derivative thermogravimetry (TG/DTG) and differential scanning calorimetry (DSC), performing optical UV-vis absorption analyses; examining the fluorescence emission-excitation matrix, total carotenoids, and color, and conducting metabolic assessments in an in vivo experimental trial. The fatty acid profile presented a higher fraction of linoleic acid (C18:2) in G and GL and alpha-linolenic acid (C18:3) in L. The acidity and peroxide indices were within the recommended ranges. The TG/DTG, DSC, and Rancimat analyses revealed similar behaviors, and the optical analyses revealed color variations caused by carotenoid contents in L and GL. In the in vivo trial, G (G2: 2000 mg/kg/day) promoted lower total consumption, and the blend (GL: 2000 mg/kg/day) group exhibited less weight gain per gram of consumed food. The group with G supplementation (G2: 2000 mg/kg/day) and GL had the highest levels of HDL-c. The group with L supplementation (L2: 2000 mg/kg/day) had the lowest total cholesterol level. The L2, G1 (1000 mg/kg/day), and G2 groups exhibited the lowest MCP-1 and TNF-α values. Additionally, the lowest adipocyte areas occurred in G and GL. Our results suggest that this combination is of high quality for consumption and can influence lipid profiles, markers of inflammation, and antioxidant status.

Water Stress Influences Phytoestrogen Levels in Red Clover (Trifolium pratense) but Not Kura Clover (T. ambiguum).

Some forage legumes synthesize phytoestrogens. We conducted a glasshouse study to investigate how water stress (drought and waterlogging) influences phytoestrogen accumulation in red clover and kura clover. Compared to the red clover control, the 20 day drought resulted in an over 100% increase in the phytoestrogens formononetin and biochanin A, which together accounted for 91-96% of the total phytoestrogens measured. Waterlogging resulted in elevated concentrations of daidzein, genistein, and prunetin but not formononetin or biochanin A. Concentrations of phytoestrogens in kura clover were low or undetectable, regardless of water stress treatment. Leaf water potential was the most explanatory single-predictor of the variation in concentrations of formononetin, biochanin A, and total phytoestrogens in red clover. These results suggest that drought-stressed red clover may have higher potential to lead to estrogenic effects in ruminant livestock and that kura clover is a promising alternative low- or no-phytoestrogen perennial forage legume.

Effects of Mixed Pasture Legume Phytoestrogens on Superovulatory Response and Embryo Quality in Angus Cows.

Ovulation and artificial insemination rates have been observed to decrease in sheep and cows when exposed to dietary phytoestrogens at concentrations greater than 25 mg/kg DM. A grazing trial was undertaken to investigate the effects of coumestrol and other key phytoestrogens on the superovulatory response, embryo numbers and quality in beef cows grazing legume pastures. A 7-week controlled grazing trial was conducted with legume and ryegrass pasture treatments, with cows exposed to legumes at two timed treatments, 4 and 7 weeks. Twenty Angus cows were subjected to a conventional estrus synchronization and superovulation protocol. Embryos were recovered via conventional uterine body flushing 7 days post artificial insemination (AI). Numerous phytoestrogens were identified in both pasture and plasma samples, including coumestrol and formononetin. Concentrations of phytoestrogens in the pasture ranged from 0.001 to 47.5 mg/kg DM and 0 to 2.6 ng/mL in plasma. Approximately 50% of cows produced viable embryos 7 days post AI. A significant interaction between the effect of treatment groups on the embryo stage was observed (p < 0.05). The results suggest that concentrations of >25 mg/kg DM of phytoestrogens less than 20 days preceding AI may negatively affect oocyte developmental competence, reduce progesterone production and thus contribute to early embryonic loss.

Effects of ethyl acetate extracts of Marsilea crenata Presl. leaves on risks of atherosclerosis in dyslipidemic rats

Background: Dyslipidemia is widely known as a significant contributor to the incidence of cardiovascular diseases (CVD). Over the years, traditional therapeutic strategies, including the use of herbs, have been recognized. Marsilea crenata Presl. (MC), or a plant known for its phytoestrogen content, has shown biological activities similar to oestrogen. These activities comprise the reduction of serum cholesterol levels, thereby having potential roles in managing dyslipidaemia and its associated cardiovascular risks.Objective: This study aims to examine the effects of ethyl acetate extracts of MC leaves on the serum lipid profile of experimental rats with dyslipidaemia induced by high-fat diet (HFD). Methods: This study applied a pretest-post-test study design with control group, involving 30 rats which are divided into 5 treatment groups, namely N (normal), HFD, STA (Simvastatin), MC1, MC2, and MC3. Group N was only treated with standard feed, and the others received HFD on days 1-14. On the 15th – 28th day, the HFD group was treated with standard feed, and the STA group received simvastatin 0.36 mg/200 g BW/day. The groups of MC1, MC2, and MC3 were treated with oral MC extracts at 100, 200, and 400 mg/kgBW, respectively. Their body weight was measured at the beginning of the study, as well as after 2 and 4 weeks. Subsequently, the experimental rats were fasted overnight on the 14th and 28th day. Their blood was collected on the 15th day (pre-test) and 29th day (post-test) to measure serum concentration of total cholesterol (TC), triglycerides (TG), and high-density lipoprotein cholesterol (HDL-c). Low-density lipoprotein cholesterol (LDL-c) concentration was also calculated by using the Friedewald formula. Results: MC3 extracts significantly reduced the TC (-40.95 ± 1.77), TG (-46.12 ± 3.79), and LDL-c levels (-0.017 ± 0.010), and it also increased levels of HDL-c (11.28 ± 2.34) mg/dL in the HFD-induced rats.Conclusion: Ethyl acetate extracts of MC leaves could reduce risk factors for atherosclerosis by returning lipid profile of the dyslipidemic rats to normal conditions with an increased dose.

Α novel method for the efficient simultaneous quantification of 67 phytoestrogens in plants and foodstuffs

The significant beneficial effects of phytoestrogens on human and animal health have emerged the necessity to develop a sensitive, accurate and rapid method for their efficient quantitation in human and animal foods. Study herein concerns the development of a novel analytical method using high-performance liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) that allows the simultaneous quantitation of 67 natural phytoestrogens. The novel method was fully validated through the determination of the following parameters: selectivity, sensibility, recovery, precision, accuracy of detection and quantification limits for each analyte. Specifically, the novel method proved accurate displaying linear response with R2 values ≥ 0.9991, quantitation limits from 5.5 to 331.8 ng/mL for 98.5% of analytes, recovery values ranging between 64.8% and 150.3%, while the precision and accuracy scores remained below 20% for most analytes. The method was successfully applied on the following plant samples used in human and/or animal diet belonging to Fabaceae plant family: Trifolium repens, Vicia sativa, Pisum sativum and Glycine max (soybean meal). Furthermore, the novel method was applied for the determination of the phytoestrogenic content of two animal feeds (for livestock and fishes) and two foodstuffs (chicken meat and fish fillet).

235 Effects of Curing Extent on the Phytoestrogen Concentrations of Red Clover Hay and Silage Across Storage Phases

Abstract Our objective was to assess the effects of insufficient (WET) or ample (CUR) curing on the phytoestrogen concentrations of red clover silage (29.4 and 45.3% DM) and hay (65.1 and 89.1, respectively). Measurements were taken at the start of storage (STRT), after 14 d (MicA), and once storage processes had stabilized for hay and silage (50 and 78 d, respectively; LATE). Hay was packed into mini-bales (~500 fresh g; ~232 kg DM/m3) and silage into mini-silos (19.5 L; ~214 kg DM/m3). Data were analyzed as a randomized complete block design (5 blocks) with a 2 (curing extents) x 2 (storage methods) x 3 (storage phases) factorial. Differences were declared at P ≤ 0.05. The concentration of formononetin was greater in silage vs. hay at STRT (4,624 vs. 2,893 ± 175 mg/kg of DM, respectively), MicA (5,383 vs. 2,110), and LATE (5,666 vs. 2,569). Also, more formononetin was found in WET vs. CUR silage (5,841 vs. 4,608.1± 154 mg/kg of DM, respectively). However, no differences were observed between WET vs. CUR hay (x̅ = 2,524). At STRT, biochanin A was greater in silage vs. hay (3,105 vs. 2,012 ± 139 mg/kg of DM, respectively). A similar trend was observed at MicA (3,233 vs. 1,057), and LATE (3,295 vs. 1,228). In addition, more biochanin A was observed in WET vs. CUR silage (3,793 vs. 2,628.9 ± 127 mg/kg of DM, respectively). The opposite was observed in WET vs. CUR hay (1,219.5 vs. 1,644.9). The genistein concentration was greater in silage vs. hay at STRT (149 vs. 120 ± 8.36 mg/kg of DM, respectively), MicA (246 vs. 62.6), and LATE (264 vs. 72.9). Also, the concentration of genistein was greater in WET vs. CUR silage (236 vs. 202.3 ± 7.21 mg/kg of DM, respectively) but the opposite was observed in WET vs. CUR hay (72.8 vs. 97.3). In the case of daidzein, hay had a greater concentration of this phytoestrogen than silage (12.8 vs. 6.67 ± 0.43 mg/kg of DM, respectively). In conclusion, red clover silage maintains greater concentrations of formononetin, biochanin A, and genistein than hay during curing period and across the storage phase. Also, adequate curing is more beneficial for silage than hay in terms of decreasing the phytoestrogen concentrations.

Association between Phytoestrogen Consumption and Female Reproductive Health: A Systematic Review of Experimental Models

AbstractPhytoestrogens have been shown as promising therapeutic agents for the treatment of menopausal symptoms, osteoporosis, breast cancer, and cardiovascular diseases. However, due to its unique chemical structure, phytoestrogen may cause unintended estrogenic and/or antiestrogenic effects on the human body, especially with regard to female reproductive health and performance. Hence, this systematic review aims to provide a critical evaluation of in vitro and in vivo evidence from the literature regarding the adverse effects of phytoestrogens on female reproductive health. The literature search was performed on four electronic databases including Scopus, Science Direct, PubMed, and Google Scholar following the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) guidelines. A total of 965 studies were screened but only 58 of them were found to be relevant and assessed for eligibility. Of these, 23 studies met the eligibility criteria while the remaining studies were excluded due to insufficiently described methods and lack of clear findings being reported. From the review, phytoestrogens may alter the development of reproductive organs, prolong the estrus cycle, induce the accumulation of fluid in the uterus, and inhibit ovulation. The concentration and exposure duration of phytoestrogens may have different effects on the reproductive organs. Thus, further studies are warranted on the toxicodynamic, toxicokinetic, mode of action, and mechanism of actions of phytoestrogens on the female reproductive system to establish recommendations regarding phytoestrogen supplement consumption for women.

Phytoestrogens Present in Follicular Fluid and Urine Are Positively Associated with IVF Outcomes following Single Euploid Embryo Transfer

The impact and safety of phytoestrogens, plant-derived isoflavones with estrogenic activity predominantly present in soy, on female reproductive health and IVF outcomes continues to be hotly debated. In this prospective cohort study, 60 women attending IVI-RMA New Jersey undergoing IVF with single frozen embryo transfer (SET/FET) of good-quality euploid blastocyst after PGT-A analysis were recruited. Concentrations of two phytoestrogens (daidzein and genistein) in follicular fluid (FF) and urine (U) were measured by UPLC–MSMS, both collected on vaginal oocyte retrieval day. These measurements correlated with IVF clinical outcomes. In models adjusted for age, BMI, race/ethnicity, and smoking status, higher FF phytoestrogen concentrations were significantly associated with higher serum estradiol, enhanced probability of implantation, clinical pregnancy, and live birth. Moreover, higher urine phytoestrogen concentrations were significantly associated with improved oocyte maturation and fertilization potential and increased probability of clinical pregnancy and live birth. Finally, higher FF and urine phytoestrogen concentrations were associated with a higher probability of live birth from a given IVF cycle. Our results suggest that dietary phytoestrogens improved reproductive outcomes of women undergoing IVF treatment. However, additional prospective studies are needed to optimize the use of phytoestrogens to further enhance reproductive outcomes and/or protect against reproductive insults.

Seasonal variation in contents of phytoestrogens in Danish dairy milk lines of different farm management systems

Farm management systems and seasonal variation are suggested to affect the concentrations of phytoestrogens in milk. The present study analyses the concentrations of phytoestrogens and their relative composition in distinct milk lines from six Danish dairies representing three management systems (conventional, organic and biodynamic) in five sampling periods (P1–P5) over one year. A highly selective and sensitive liquid chromatography-triple quadrupole mass spectrometry method was applied to determine absolute concentrations of phytoestrogens in the different milk lines over the period. Biodynamic dairy milk lines contained higher levels of most isoflavones compared with milk from conventional systems. This may be explained by the differences in feeding strategies and, more specifically, botanical composition of feed. In relation to season, the concentration of the mammalian derived isoflavone, equol, was highest during the indoor period in biodynamic milk. Compared with the isoflavones, concentrations of lignans were less affected by farm management system.

Phytoestrogens as Biomarkers of Plant Raw Materials Used for Fish Feed Production.

The intensive use of plant materials as a sustainable alternative for fish feed production, combined with their phytochemical content, which affects the growth and production characteristics of farmed fishes, necessitates their monitoring for the presence of raw materials of plant origin. This study reported herein concerns the development, validation and application of a workflow using high-performance liquid chromatography combined with tandem mass spectrometry (LC-MS/MS) for the quantification of 67 natural phytoestrogens in plant-derived raw materials that were used to produce fish feeds. Specifically, we verified the presence of 8 phytoestrogens in rapeseed meal samples, 20 in soybean meal samples, 12 in sunflower meal samples and only 1 in wheat meal samples in quantities enabling their efficient incorporation into clusters. Among the various constituents, the soybean phytoestrogens daidzein, genistein, daidzin, glycitin, apigenin, calycosin and coumestrol, as well as the sunflower neochlorogenic, caffeic and chlorogenic phenolic acids, displayed the highest correlations with their origin descriptions. A hierarchical cluster analysis of the studied samples, based on their phytoestrogen contents, led to the efficient clustering of raw materials. The accuracy and efficiency of this clustering were tested through the incorporation of additional samples of soybean meal, wheat meal and maize meal, which verified the utilization of the phytoestrogen content as a valuable biomarker for the discrimination of raw materials used for fish feed production.

Effects on Serum Hormone Concentrations after a Dietary Phytoestrogen Intervention in Patients with Prostate Cancer: A Randomized Controlled Trial.

Phytoestrogens have been suggested to have an anti-proliferative role in prostate cancer, potentially by acting through estrogen receptor beta (ERβ) and modulating several hormones. We primarily aimed to investigate the effect of a phytoestrogen intervention on hormone concentrations in blood depending on the ERβ genotype. Patients with low and intermediate-risk prostate cancer, scheduled for radical prostatectomy, were randomized to an intervention group provided with soybeans and flaxseeds (∼200 mg phytoestrogens/d) added to their diet until their surgery, or a control group that was not provided with any food items. Both groups received official dietary recommendations. Blood samples were collected at baseline and endpoint and blood concentrations of different hormones and phytoestrogens were analyzed. The phytoestrogen-rich diet did not affect serum concentrations of testosterone, insulin-like growth factor 1, or sex hormone-binding globulin (SHBG). However, we found a trend of decreased risk of increased serum concentration of estradiol in the intervention group compared to the control group but only in a specific genotype of ERβ (p = 0.058). In conclusion, a high daily intake of phytoestrogen-rich foods has no major effect on hormone concentrations but may lower the concentration of estradiol in patients with prostate cancer with a specific genetic upset of ERβ.

Preparation of pumpkin oil-based nanoemulsion as a potential estrogen replacement therapy to alleviate neural-immune interactions in an experimental postmenopausal model

As estrogen production decreases during menopause; the brain's metabolism tends to stall and become less effective. Estrogen most likely protects against neurodegeneration. Consequently, a comprehensive study of the benefits of hormone replacement therapy as a neuroprotective effect is urgently required. This study was designed to fabricate pumpkin seed oil nanoparticles (PSO) in nanoemulsion form (PSO-NE) and investigate their potential role in attenuating the neural-immune interactions in an experimental postmenopausal model.Sixty female white albino rats were divided into six groups: control, sham, ovariectomized (OVX), and three OVX groups treated with 17β-estradiol, PSO, and PSO-NE respectively. Transmission Electron Microscopy (TEM), and particle size analyzer were performed for nanoemulsion evaluation. Serum levels of estrogen, brain amyloid precursor protein (APP), serum levels of nuclear factor kappa B (NF-κβ), interleukin 6 (IL-6), transthyretin (TTR), and synaptophysin (SYP) were evaluated. The expression of estrogen receptors (ER-α, β) in the brain tissue was estimated. The findings revealed that the approached PSO-NE system was able to reduce the interfacial tension, enhance the dispersion entropy, lower the system free energy to an extremely small value, and augment the interfacial area. PSO-NE, showed a significant increase in the levels of estrogen, brain APP, SYP, and TTR accompanied with a significant increased in the expression of brain ER-α, β compared to the OVX group. In conclusion, the phytoestrogen content of PSO exhibited a significant prophylactic effect on neuro-inflammatory interactions, ameliorating both estrogen levels and the inflammatory cascades.

In utero and postnatal exposure to Foeniculum vulgare and Linum usitatissimum seed extracts: modifications of key enzymes involved in epigenetic regulation and estrogen receptors expression in the offspring’s ovaries of NMRI mice

BackgroundEarly-life exposure to exogenous estrogens such as phytoestrogens (plant-derived estrogens) could affect later health through epigenetic modifications. Foeniculum vulgare (fennel) and Linum usitatissimum (flax) are two common medicinal plants with high phytoestrogen content. Considering the developmental epigenetic programming effect of phytoestrogens, the main goal of the present study was to evaluate the perinatal exposure with life-long exposure to hydroalcoholic extracts of both plants on offspring’s ovarian epigenetic changes and estrogen receptors (ESRs) expression level as signaling cascades triggers of phytoestrogens.MethodsPregnant mice were randomly divided into control (CTL) that received no treatment and extract-treated groups that received 500 mg/kg/day of fennel (FV) and flaxseed (FX) alone or in combination (FV + FX) during gestation and lactation. At weaning, female offspring exposed to extracts prenatally remained on the maternal-doses diets until puberty. Then, the ovaries were collected for morphometric studies and quantitative real-time PCR analysis.ResultsA reduction in mRNA transcripts of the epigenetic modifying enzymes DNMTs and HDACs as well as estrogen receptors was observed in the FV and FX groups compared to the CTL group. Interestingly, an increase in ESRα/ESRβ ratio along with HDAC2 overexpression was observed in the FV + FX group.ConclusionOur findings clearly show a positive relationship between pre and postnatal exposure to fennel and flaxseed extracts, ovarian epigenetic changes, and estrogen receptors expression, which may affect the estrogen signaling pathway. However, due to the high phytoestrogen contents of these extracts, the use of these plants in humans requires more detailed investigations.

Comprehensive compositional assessment of bioactive compounds in diverse pea accessions

Pea (Pisum sativum L.) is an important legume crop providing a good source of protein, vitamins, minerals and bioactive compounds with health benefits for humans. In this study, an improved method for simultaneous analysis of multiple phytoestrogens among 100 pea accessions was developed. Ipriflavone, (a synthetic isoflavone), was used as an internal standard for the semiquantitative analysis of 17 phytoestrogens including isoflavone aglycones and conjugates, allowing direct analysis of isoflavones in their naturally occurring forms. This comprehensive dataset demonstrated that the isoflavones varied greatly and some accessions tended to have high levels of multiple phytoestrogens among the 100 accessions analyzed. Isoliquiritigenin followed by glycitein were the predominant compounds detected in the accessions and showed the highest correlation with the total phytoestrogens content. Secoisolariciresinol content was consistently higher in yellow cotyledon peas than in green cotyledon peas, whereas the contents of coumestrol, genestein and secoisolariciresinol were significantly correlated with seed coat color. The total phenolics and saponins showed a wide range of variability among the accessions with higher concentrations of total phenolics observed in seeds with pigmented seed coat or yellow cotyledon seeds, suggesting the synthesis of saponins and phenolics are significantly affected by metabolic pathway genes controlling cotyledon color or seed coat color. This study profiled the variability of bioactive compounds of pea seed quality traits in diverse pea accessions and provides an immense resource for continued research, breeding and selection of genotypes for a wide range of applications.

Novel cord blood and urinary phytoestrogens levels in male neonates with normal external genitalia

Objective: This was a new prospective study to determine the association between phytoestrogen levels in the cord blood and urine with normal and abnormal external genitalia in male newborns. Materials and Methods: One hundred and fifty-one term male newborns were enrolled. Cord blood and urine and medical photos of their external genitalia were obtained. Prematurity and multiple congenital anomalies or syndromic characteristics were excluded. Serum and urine levels of phytoestrogens (daidzein and genistein) were calculated using liquid chromatography/mass spectrometry. The normality of the appearance of the external genitalia of the males was determined by a consensus review by experts. Results: A total of 146 cord blood and 96 newborn urine samples were obtained; only two newborns had abnormal external genitalia. In the 144 babies with normal external genitalia, mean levels were determined to be as follows; serum daidzein 10.82 ng/ml, serum genistein 42.17 ng/ml, urine daidzein 69.91 ng/ml, and urine genistein 126.09 ng/ml. In the two babies with abnormal external genitalia, mean levels of each of the measured phytoestrogen were lower; however, there was no statistical significance between the two groups (P > 0.05). Conclusions: We successfully determined and developed a novel database on the levels of measurable phytoestrogens in serum and urine from male newborns with normal external genitalia. In the normal group, a higher concentration of serum and urinary phytoestrogens was correlated with the extension of the scrotal pigmentation above and proximal to the base of the penis. These data are useful to better understand the role of phytoestrogens in the development of male genitalia and for future research on newborns with abnormal external genitalia.

Phytoestrogens: A Review of Their Impacts on Reproductive Physiology and Other Effects upon Grazing Livestock.

Simple SummaryPhytoestrogens are secondary plant metabolites that play a role in plant defense, and when ingested by livestock have numerous functions related to reproduction, metabolism, immunological functions and livestock growth and performance. Phytoestrogens are found across various plant species, with the most biologically active of these, isoflavones and coumestans, abundant in legume species. Understanding the overall potential health and reproductive effects that may occur in livestock grazing phytoestrogenic pastures is essential to mitigate any potential risks attributed to fertility loss or to introduce proactive management strategies to aid in improving growth and development. A review of their interactions with livestock systems will provide updated information for the agricultural and veterinary industries.Legume crops and pastures have a high economic value in Australia. However, legume species commonly used for grazing enterprises have been identified to produce high concentrations of phytoestrogens. These compounds are heterocyclic phenolic, and are similar in structure to the mammalian estrogen, 17β-estradiol. The biological activity of the various phytoestrogen types; isoflavones, lignans and coumestans, are species-specific, although at concentrations of 25 mg/kg of dry matter each of the phytoestrogen types affect reproductive functions in grazing livestock. The impacts upon fertility in grazing livestock such as cattle and sheep, vary greatly over length of exposure time, age and health of animal and the stress stimuli the plant is exposed to. More recently, research into the other effects that phytoestrogens may have upon metabolism, immune capacity and growth and performance of grazing livestock has been conducted. Potential new benefits for using these phytoestrogens, such as daidzein and genistein, have been identified by observing the stimulation of production in lymphocytes and other antibody cells. Numerous isoflavones have also been recognized to promote protein synthesis, increase the lean meat ratio, and increase weight gain in cattle and sheep. In Australia, the high economic value of legumes as pasture crops in sheep and cattle production enterprises requires proactive management strategies to mitigate risk associated with potential loss of fertility associated with inclusion of pasture legumes as forages for grazing livestock.

Reproductive disruption in adult female and male rats prenatally exposed to mesquite pod extract or daidzein

Phytoestrogens are considered to be endocrine disruptors, since they can alter the endocrine system, thus disturbing many reproductive events. The intake of diets containing a high content of phytoestrogens has increased worldwide in human populations and in domestic animals. Phytoestrogens in maternal blood can pass through the placenta to the fetus in high amounts and can have long-term organizational effects. Mesquite (Prosopis sp) is a leguminous plant widely used to feed several livestock species, and is also used in the human diet. In this study we assessed the effects of exposure to mesquite pod extract during the periconception and pregnancy periods on the reproduction of male and female descendants. The females of three experimental groups received one of the following treatments: 1) vehicle injection; 2) mesquite pod extract or 3) the isoflavone daidzein during the periconception and pregnancy periods. Estrous cyclicity, sexual behavior and hormones, as well as uterine and vaginal epithelia were evaluated in the female descendants. In the males, sexual behavior and hormones, apoptosis in testicular cells and sperm quality were evaluated. In females the following was observed: alterations in estrous cycles, decreased sexual behavior, estradiol and progesterone levels, increased uterine and vaginal epithelia. In males, we observed a decrease in sexual behavior, testosterone and sperm quality, and apoptosis increased in testicular cells. All these effects were similar to those caused by daidzein. These results indicate that prenatal exposure to mesquite pod extract or daidzein, administered to females before and during pregnancy, can disrupt normal organizational-activational programming of reproductive physiology in female and male descendants.