Higenamine is a catecholic benzyltetrahydroisoquinoline alkaloid occurring in plant-derived foods and food supplements, the latter potentially resulting in substantially higher exposure levels than conventional foods. Because of its catechol and alkylphenol structures, oxidative conversion to electrophilic quinone or quinone methide intermediates is plausible, raising concern about a potential genotoxic hazard. However, experimental data on the genotoxic potential of higenamine are lacking. The present study, therefore, investigated its potential to induce gene mutations and chromosomal damage in cultured V79 cells. Gene mutations were examined at the hypoxanthine-guanine phosphoribosyltransferase (HPRT) locus after 24 h treatment without metabolic activation and after 4 h in the absence or presence of rat liver S9-mix. Clastogenicity and aneugenicity were assessed using in vitro micronucleus assay with CREST staining after 4 h of treatment, followed by compound-free postincubation periods of 16 and 24 h. Proliferation and mitotic activity were monitored, and higenamine stability was analyzed by HPLC-UV. Higenamine caused a statistically significant increase in the frequency of micronuclei at 300-510 µM, reaching maxima of 51 and 73 micronucleated cells per 1000 cells without and with metabolic activation, respectively. Micronuclei mainly contained chromosomal fragments, indicating a prevalent clastogenic mode of action, although a smaller increase in micronuclei containing whole chromosomes was also observed. Mutant frequencies in the HPRT assay were not significantly increased under any condition tested, despite concentration-dependent cytotoxicity. During incubation, higenamine concentrations decreased while only minor formation of O-methylated metabolites was detected, suggesting conversion into additional reactive products. Overall, these findings provide the first experimental evidence that higenamine exhibits clastogenic potential in vitro and support the hypothesis that oxidative activation of its catechol and/or alkylphenol moiety to electrophilic intermediates capable of reacting with cellular nucleophiles might contribute to its genotoxic activity.

Green seaweed (Ulva sp.) is frequently used as a food component and nutraceutical agent because of its high polysaccharide and natural fiber content in Asian countries. This study investigates both metabolomic profiling of Ulva sp. and the neuroprotective efficacy of its ethanol extract and its underlying mechanisms in a rotenone-induced rat model of neurodegeneration, mimicking Parkinson's disease (PD) in humans. Metabolomic profiling of Ulva sp. extract was done using liquid chromatography high resolution electrospray ionization mass spectrometry and led to the identification of 22 compounds belonging to different chemical classes.Catenin Beta Additionally, this study demonstrated the neuroprotective properties against rotenone-induced PD, which was achieved through the suppression of elevated levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-6 together with the inhibition of reactive oxygen species (ROS) generation, apoptosis, inflammatory mediators, and the phosphoinositide 3-kinases/serine/threonine protein kinase (PI3K/AKT) pathway. Using a protein-protein interaction network, AKT1, GAPDH, TNF-α, IL-6, caspase 3, signal transducer and activator of transcription 3, Catenin Beta 1, epidermal growth factor receptor, B-cell lymphoma -2, and HSP90AA1 were identified as the top 10 most significant genes. Finally, molecular docking results showed that compounds 1, 3, and 7 might possess a promising anti-parkinsonism effect by binding to active sites of selected hub genes. Therefore, it is hypothesized that the Ulva sp. extract has the potential to be further developed as a potential therapeutic agent for the treatment of PD.

LebensmittelchemieVolume 78, Issue S1 p. S1-030-S1-030 Abstract The food cons tuent higenamine exhibits no genotoxic poten al in Ishikawa cells but slightly reduces the sulfotransferase-induced genotoxicity of 1-hydroxymethylpyrene Eva Veh, Eva Veh Julius-Maximilians-Universität, Lehrstuhl für Lebensmi elchemie Am Hubland, 97074 WürzburgSearch for more papers by this authorFranka Schulze, Franka Schulze Julius-Maximilians-Universität, Lehrstuhl für Lebensmi elchemie Am Hubland, 97074 WürzburgSearch for more papers by this authorLeane Lehmann, Leane Lehmann Julius-Maximilians-Universität, Lehrstuhl für Lebensmi elchemie Am Hubland, 97074 WürzburgSearch for more papers by this author Eva Veh, Eva Veh Julius-Maximilians-Universität, Lehrstuhl für Lebensmi elchemie Am Hubland, 97074 WürzburgSearch for more papers by this authorFranka Schulze, Franka Schulze Julius-Maximilians-Universität, Lehrstuhl für Lebensmi elchemie Am Hubland, 97074 WürzburgSearch for more papers by this authorLeane Lehmann, Leane Lehmann Julius-Maximilians-Universität, Lehrstuhl für Lebensmi elchemie Am Hubland, 97074 WürzburgSearch for more papers by this author First published: 01 March 2024 https://doi.org/10.1002/lemi.202452020AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat No abstract is available for this article. Volume78, IssueS1Supplement: Abstracts der Vorträge der Regionalverbände und die der Posterflashtalks der AG JLCMarch/April 2024Pages S1-030-S1-030 RelatedInformation

Gladiolus segetum Ker-Gawl corms total extract exhibited remarkable in vitro anti-proliferative effects against panel of cancer cell lines; including human colon carcinoma (Caco-2), human breast cancer (MCF7) and hepatocellular carcinoma (HepG2) cell lines with IC50 values of 7.4, 9.1 and 11.2 μg/ml, respectively. The total ethanolic extract of G. segetum Ker-Gawl corms was subjected to untargeted metabolomics profiling using LC-HR-ESI-MS, which revealed the presence of various clusters of phytoconstituents as triterpenes, anthraquinones, flavonoids and phenolic derivatives. Network pharmacology study was performed for all identified compounds, the formed networks identified 73 intersected genes. The diagrammatic illustration of the top pathways revealed that phosphoinositide 3-kinase (PI3 K) gene is the effective dominant gene in the top four KEGG pathways. Upon molecular docking and molecular dynamics investigation, kaempferol-3-O-glucopyranoside was suggested to be key anticancer metabolite. Interestingly, cytotoxic investigation of this compound revealed potential activity against the tested cancer cell lines (Caco-2, MCF7 and HepG2) with IC50 values of 6.2, 8.5 and 9.3 μg/ml, respectively. The present study highlighted the potential of G. segetum Ker-Gawl as a promising source of interesting anticancer scaffolds.

When investigating endocrine disorders, it is essential to assess a comprehensive quantitative profile of sex (pro)hormones in plasma including conjugates. Thus, the present study aimed to develop and validate a comprehensive mass spectrometry-based multimethod combining the direct analysis of unconjugated sex (pro)hormones and oxidation products thereof (by GC), as well as their sulfates and glucuronides present in higher concentrations (by LC) with the indirect quantification of glucuronides present in lower concentrations after selective glucuronide hydrolysis (by GC) and its application to plasma derived from ten pre- and postmenopausal women and men each. Even guideline-compliant validation experiments cannot completely reflect overestimation of analyte concentrations due to effects depending on the individual ratio of analytes (i.e. chemical formation of analytes or incomplete removal of interfering analytes). Thus, the extent of processes not accounted for by the calibration strategy were investigated and maximum over- or underestimations of analyte concentrations were assessed for each plasma sample individually. 34 analytes were successfully calibrated, validated (median accuracy 101.1 %, median inter-day precision 8.1 %) and 31 were detected above the detection limit in plasma samples. The sporadic maximum individual over- or underestimation of analyte concentrations amounted to less than 20 %.

LebensmittelchemieVolume 77, Issue S2 p. S2-157-S2-157 Julius-Maximilians-Universität Würzburg Application based personalized food choices and health sustainment: scientific background and investigation of biomarkers in human tissue specimens Tobias Jaud, Tobias JaudSearch for more papers by this authorBetreuerin: Prof. Dr. Leane Lehmann, Betreuerin: Prof. Dr. Leane LehmannSearch for more papers by this author Tobias Jaud, Tobias JaudSearch for more papers by this authorBetreuerin: Prof. Dr. Leane Lehmann, Betreuerin: Prof. Dr. Leane LehmannSearch for more papers by this author First published: 01 June 2023 https://doi.org/10.1002/lemi.202352262AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL No abstract is available for this article. Volume77, IssueS2June 2023Pages S2-157-S2-157 RelatedInformation

Searchable abstracts of presentations at key conferences in endocrinology ISSN 1470-3947 (print) | ISSN 1479-6848 (online)

In epidemiological studies, blood levels of 17β-estradiol (E2) are associated with hormone-dependent diseases. The lack of specific methods impedes studies on the role of E2 metabolites and their conjugates in the etiology of hormone-dependent diseases. Stable-isotope dilution tandem mass spectrometry methods (coupled to gas chromatography and liquid chromatography systems) for the analysis of 22 endogenous estrogens, including both oxidative metabolites, as well as sulfates and glucuronides, was validated and the method applied to plasma of women with no breast cancer. No changes in estrogen profile during sample cleanup were observed and values for limit of detection (7fmol/ml - 2 pmol/ml), accuracies (80-122%) as well as intra- and inter-day precision (below 28%) at levels near the limit of quantification were acceptable. In human plasma only seven estrogens were detected and estrone conjugates contributed most to the estrogen profile.

Breast cancer etiology is associated with both proliferation and DNA damage induced by estrogens. Breast cancer risk factors (BCRF) such as body mass index (BMI), smoking, and intake of estrogen-active drugs were recently shown to influence intratissue estrogen levels. Thus, the aim of the present study was to investigate the influence of BCRF on estrogen-induced proliferation and DNA damage in 41 well-characterized breast glandular tissues derived from women without breast cancer. Influence of intramammary estrogen levels and BCRF on estrogen receptor (ESR) activation, ESR-related proliferation (indicated by levels of marker transcripts), oxidative stress (indicated by levels of GCLC transcript and oxidative derivatives of cholesterol), and levels of transcripts encoding enzymes involved in estrogen biotransformation was identified by multiple linear regression models. Metabolic fluxes to adducts of estrogens with DNA (E-DNA) were assessed by a metabolic network model (MNM) which was validated by comparison of calculated fluxes with data on methoxylated and glucuronidated estrogens determined by GC- and UHPLC-MS/MS. Intratissue estrogen levels significantly influenced ESR activation and fluxes to E-DNA within the MNM. Likewise, all BCRF directly and/or indirectly influenced ESR activation, proliferation, and key flux constraints influencing E-DNA (i.e., levels of estrogens, CYP1B1, SULT1A1, SULT1A2, and GSTP1). However, no unambiguous total effect of BCRF on proliferation became apparent. Furthermore, BMI was the only BCRF to indeed influence fluxes to E-DNA (via congruent adverse influence on levels of estrogens, CYP1B1 and SULT1A2).

In Eastern Asia, coronary heart disease as well as breast and prostate cancer are less frequent than in Western countries, which is considered to be due to the Asian diet. Because Asians consume much more soy-based food products, putative beneficial health effects of soy and soy isoflavones have attracted much attention in the past two decades. Yet, although a plethora of cell-free studies, studies in vitro and in vivo, and studies in humans have investigated a multitude of putatively beneficial biological effects, no health claim directly related to isoflavones has been approved up to now. Likewise, despite ongoing safety evaluation of isoflavones by scientific expert panels, no conclusion is expected to be released in the near future. The present chapter summarizes the current knowledge on toxicity and efficacy of isoflavones, focusing on biokinetic properties of isoflavones necessary to understand the heterogeneity of study results and other difficulties in evaluating safety and efficacy of isoflavones.

Understanding intramammary estrogen homeostasis constitutes the basis of understanding the role of lifestyle factors in breast cancer etiology. Thus, the aim of the present study was to identify variables influencing levels of the estrogens present in normal breast glandular and adipose tissues (GLT and ADT, i.e., 17β-estradiol, estrone, estrone-3-sulfate, and 2-methoxy-estrone) by multiple linear regression models. Explanatory variables (exVARs) considered were (a) levels of metabolic precursors as well as levels of transcripts encoding proteins involved in estrogen (biotrans)formation, (b) data on breast cancer risk factors (i.e., body mass index, BMI, intake of estrogen-active drugs, and smoking) collected by questionnaire, and (c) tissue characteristics (i.e., mass percentage of oil, oil%, and lobule type of the GLT). Levels of estrogens in GLT and ADT were influenced by both extramammary production (menopausal status, intake of estrogen-active drugs, and BMI) thus showing that variables known to affect levels of circulating estrogens influence estrogen levels in breast tissues as well for the first time. Moreover, intratissue (biotrans)formation (by aromatase, hydroxysteroid-17beta-dehydrogenase 2, and beta-glucuronidase) influenced intratissue estrogen levels, as well. Distinct differences were observed between the exVARs exhibiting significant influence on (a) levels of specific estrogens and (b) the same dependent variables in GLT and ADT. Since oil% and lobule type of GLT influenced levels of some estrogens, these variables may be included in tissue characterization to prevent sample bias. In conclusion, evidence for the intracrine activity of the human breast supports biotransformation-based strategies for breast cancer prevention. The susceptibility of estrogen homeostasis to systemic and tissue-specific modulation renders both beneficial and adverse effects of further variables associated with lifestyle and the environment possible.