Data on the optimisation of a solid phase extraction method for fractionating estrogen metabolites from small urine volumes

Berg, Carien L. van der; Venter, Gerda; Westhuizen, Francois H. van der; Erasmus, Elardus

doi:10.1016/j.dib.2020.105222

Cited by 4 publications

(8 citation statements)

References 3 publications

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“…For the glucuronidation of E3, only two glucuronides (E 3 -3O-G and E 3 -16O-G) were detected in HLM or HIM incubation Frontiers in Pharmacology frontiersin.org systems (Figure 7), which was consistent with the previous study (Sneitz et al, 2013). In addition, several research studies have shown that E 1 -3O-G, E 2 -3O-G, and E 3 -16O-G were the main glucuronide conjugates of E 1 , E 2, and E 3 in urine, respectively (Tikkanen 1973;Van der Berg et al, 2020). Also, only E 1 -3O-G and E 2 -3O-G could be detected in postmenopausal women's serum (Caron et al, 2009), which kept in line with our resulting in vitro glucuronidation assays (Figure 7).…”

Section: Discussionsupporting

confidence: 89%

Identification of naturally occurring inhibitors in Xian-Ling-Gu-Bao capsule against the glucuronidation of estrogens

Wang

et al. 2022

Front. Pharmacol.

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Xian-Ling-Gu-Bao (XLGB) capsule, a well-known traditional Chinese medicine prescription, is widely used for the treatment of osteoporosis. It could significantly increase the levels of estrogen in ovariectomized rats and mice. However, this working mechanism has not been well elucidated. Considering that UDP-glucuronosyltransferase (UGT) enzymes are the important enzymes that inactivate and regulate estrogen activity in vivo, this study aimed to identify the bioactive compounds from XLGB against the glucuronidation of estrogens. First, thirty compounds were considered as candidate bioactive compounds based on our previous studies including pharmacological evaluation, chemical profiles, and metabolic profiles. Second, the characteristics of estrogen glucuronidation by uridine diphosphate glucuronic acid (UDPGA)-supplemented human liver microsomes (HLM), human intestine microsomes (HIM), and expressed UGT enzymes were determined, and the incubation systems of their key UGT enzymes were optimized. Then, inhibitory effects and mechanisms of XLGB and its main compounds toward the key UGT isozymes were further investigated. As a result, estrogen underwent efficient glucuronidation by HLM and HIM. UGT1A10, 1A1, and 2B7 were mainly responsible for the glucuronidation of estrone, β-estradiol, and estriol, respectively. For E1 and E2, UGT1A10 and 1A1 tended to mediate estrogen-3-O-glucuronidation, while UGT2B7 preferred catalyzing estrogen-16-O-glucuronidation. Furthermore, the incubation system for active UGT isoforms was optimized including Tris-HCl buffer, detergents, MgCl2 concentration, β-glucuronidase inhibitors, UDPGA concentration, protein concentration, and incubation time. Based on optimal incubation conditions, eleven, nine, and nine compounds were identified as the potent inhibitors for UGT1A10, 1A1, and 2B7, respectively (IC50 < 4.97 μM and Ki < 3.35 μM). Among them, six compounds (bavachin, isobavachin, isobavachalcone, neobavaisoflavone, corylifol A, and icariside II) simultaneously demonstrated potent inhibitory effects against these three active enzymes. Prenylated flavanols from Epimedium brevicornu Maxim., prenylated flavonoids from Psoralea corylifolia L., and salvianolic acids from Salvia miltiorrhiza Bge. were characterized as the most important and effective compounds. The identification of potent natural inhibitors of XLGB against the glucuronidation of estrogen laid an important foundation for the pharmacodynamic material basis.

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Section: Discussionsupporting

confidence: 89%

Identification of naturally occurring inhibitors in Xian-Ling-Gu-Bao capsule against the glucuronidation of estrogens

Wang

et al. 2022

Front. Pharmacol.

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“…We hypothesized that the vEHT may differ from oral and transdermal patch in composition, metabolism, or dosage. We therefore used a modified version of previously published targeted liquid-chromatography mass spectrometry (LC-MS) methods for the measurement of urinary estrogens to quantify excreted urinary estrogen conjugates of women in the No UTI History and rUTI Remission groups (van der Berg et al, 2020). Limiting our analysis to the known major excreted sulfate and glucuronide conjugates of estrone (E1) and 17b-estradiol (E2), we observed significantly higher urinary E1 and E2 conjugate abundance, both sulfates and glucuronides, in women using oral EHT, an observation concordant with observed urinary Lactobacillus abundance (Figure 5 We further sought to determine if rUTI history differentially affected the EHT-associated taxonomic signature by dichotomizing patients by No UTI History and rUTI Remission group membership.…”

Section: Lactobacillus Abundance In the Urobiome Of Postmenopausal Womenmentioning

confidence: 99%

“…We hypothesized that vEHT may differ from oral and transdermal patch in composition, metabolism, or dosage. We therefore optimized a published targeted liquid-chromatography mass spectrometry (LC-MS) method to quantify excreted urinary estrogen conjugates of women in the No UTI History and rUTI Remission groups (van der Berg et al, 2020). Limiting our analysis to the known major excreted sulfate and glucuronide conjugates of estrone (E1) and 17b-estradiol (E2), we observed significantly higher urinary concentrations of E1 and E2 sulfates and glucuronides in women using oral EHT.…”

Section: Urinary Estrogen Concentration Is Positively Correlated With Urobiome Lactobacillus Abundance In Pm Women With No Uti Historymentioning

confidence: 99%

“…previously reported methods(van der Berg et al, 2020). Briefly, urine (500 µL) was diluted and spiked with 100 ng stable isotope-labeled internal standards of d3-Estrone 3-Glucuronide and d4-Estradiol 3-Sulfate.…”

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confidence: 99%

“…Plates were incubated inverted per the recommendations of Clinical and Laboratory Standards Institute (CLSI) M100-ED30: 2020 Performance Standards for AntimicrobialSusceptibilityTesting, 30 th Edition (https://clsi.org/standards/products/microbiology/documents/m100/). After incubation, antimicrobial susceptibility was evaluated by measurement of the zone of inhibition and using CLSI established zone diameter breakpoints.Liquid Chromatography Mass Spectrometry Measurement of Estrogen MetabolitesDirect measurement of urinary estrogen metabolites was performed via a modification of previously reported methods(van derBerg et al, 2020). Briefly, urine (500 µL) was diluted and spiked with 100 ng stable isotope-labeled internal standards of d3-Estrone 3-Glucuronide and d4-Estradiol 3-Sulfate.…”

mentioning

confidence: 99%

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Recurrent urinary tract infection and estrogen shape the taxonomic ecology and functional potential of the postmenopausal urobiome

Neugent

Kumar

Hulyalkar

et al. 2021

Preprint

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Community-acquired urinary tract infection (UTI) is among the most common bacterial infections observed in humans. Postmenopausal women are a rapidly growing and underserved demographic group who are severely affected by rUTI with a >50% recurrence rate. In this population, rUTI can persist for years, reducing quality of life and imposing a significant healthcare burden. rUTI is most often treated by long-term antibiotic therapy, but development of antibiotic resistance and allergy leave physicians with fewer treatment options. The female urobiome has been identified as a key component of the urogenital environment. However, structural and functional changes in the urobiome underlying rUTI susceptibility in postmenopausal women are not well understood. Here, we used strictly curated, controlled cross-sectional human cohorts of postmenopausal women, urobiome whole genome (shotgun) metagenomic sequencing (WGMS), advanced urine culturing techniques, extensive biobanking of >900 patient-derived urinary bacterial and fungal isolates, and mass spectrometry-based estrogen profiling to survey the urobiome in rUTI patients during infection relapse and remission as well as healthy comparators with no lifetime history of UTI. Our results suggest that a history of rUTI strongly shapes the taxonomic and functional ecology of the urobiome. We also find a putative protective commensal population, consisting of species known to convey protection against bacterial vaginosis such as Lactobacillus crispatus, within the urobiome of women who do not experience UTI. Integration of clinical metadata detected an almost exclusive enrichment of putative protective species belonging to the genus, Lactobacillus, in women taking estrogen hormone therapy (EHT). We further show that the urobiome taxonomic ecology is shaped by EHT, with strong enrichments of putatively protective lactobacilli, such as L. crispatus and L. vaginalis. Integrating quantitative metabolite profiling of urinary estrogens with WGMS, we observed robust associations between urobiome taxa, such as Bifidobacterium breve and L. crispatus, and urinary estrogen conjugate concentrations, suggesting that EHT strongly alters the taxonomic composition of the female urobiome. We have further used functional metagenomic profiling and patient-derived isolate phenotyping to identify microbial metabolic pathways, antimicrobial resistance genes (ARGs), and clinically relevant antimicrobial resistance phenotypes enriched between disease-states. Our data suggest distinct metabolic and ARG signatures of the urobiome associated with current rUTI status and history. Taken together, our data suggests that rUTI history and estrogen use strongly shape the functional and taxonomic composition of the urobiome in postmenopausal women.

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Steroid determination—Purification of extracts

Honour

2023

Steroids in the Laboratory and Clinical Practice

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Data on the optimisation of a solid phase extraction method for fractionating estrogen metabolites from small urine volumes

Cited by 4 publications

References 3 publications

Identification of naturally occurring inhibitors in Xian-Ling-Gu-Bao capsule against the glucuronidation of estrogens

Identification of naturally occurring inhibitors in Xian-Ling-Gu-Bao capsule against the glucuronidation of estrogens

Recurrent urinary tract infection and estrogen shape the taxonomic ecology and functional potential of the postmenopausal urobiome

Steroid determination—Purification of extracts

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