Journal: bioRxiv

Article Title: Gut microbiota gate host exposure to metabolites from dietary Solanums

doi: 10.1101/2024.03.20.584512

Figure Lengend Snippet: A) Tomatine and tomatidine concentrations in serum (nM), cecal contents (pmol/mg fresh weight), and liver (pmol/mg fresh weight) following a 10% tomato dietary intervention in C57BL/6J mice receiving antibiotics (Abx). Values shown are the mean±SD for each group, with individual replicates overlaid (n=5 mice per treatment). P values are determined using a two-sided Welch’s T test. ns = not significant (P > 0.05). B) Tomatine-derived metabolites in tissues following dietary intervention with 10% tomato diet in C57BL/6J mice receiving antibiotics. Abundances for each metabolite are normalized to the maximum EIC peak area across tissues shown in bars. Peak areas were normalized by sample dry mass to enable comparison between different tissue types. Values shown are the mean for each treatment group (n=5 mice per treatment). Structures as drawn are hypothesized based on m/z and MS/MS spectra; the position of hydroxylation is not known. For molecules with multiple detected isomers, the most abundant isomer is shown. C) Chaconine, solanine, and solanidine concentrations in serum (nM), cecal contents (pmol/mg fresh weight), and liver (pmol/mg fresh weight) following a 5% potato dietary intervention in C57BL/6J mice receiving antibiotics. Values shown are the mean±SD for each treatment group, with individual replicates overlaid (n=5 mice per treatment). P values are determined using a two-sided Welch’s T test. ns = not significant (P > 0.05). D) Solanidine and chaconine in human urine from colectomy patients (n=12 donors) and donors with intact colons (n=17 donors). Compound abundance is shown as EIC peak area normalized to the creatinine content in that sample. Values shown are for individual donor samples, with the molar concentration of solanidine in one intact colon sample shown in pmol solanidine per mg creatinine.

Article Snippet: Steroidal alkaloid compounds were acquired from commercial sources: tomatine (Tokyo Chemical Industry), tomatidine (Cayman Chemical Company), solanine (Sigma Aldrich), chaconine (Sigma Aldrich), solanidine (Carbosynth), solamargine (MedChemExpress), solasonine (MedChemExpress), solasodine (Santa Cruz Biotechnology).

Techniques: Derivative Assay, Comparison, Tandem Mass Spectroscopy, Concentration Assay

Journal: bioRxiv

Article Title: Gut microbiota gate host exposure to metabolites from dietary Solanums

doi: 10.1101/2024.03.20.584512

Figure Lengend Snippet: A) Total steroidal alkaloid concentration in the serum of C57BL/6J mice fed potato-supplemented diet and treated with antibiotics. Aglycone concentrations were estimated using a solanidine standard curve. Data shown are the mean±SD (n=5 mice). P values were determined using a two-sided Welch’s T test. B) In vitro inhibition of recombinant human AChE activity by potato steroidal alkaloids. Data shown are normalized to AChE treated with vehicle only (DMSO for solanine, chaconine and ethanol for solanidine) with a total vehicle concentration of 1% for all treatments. Values shown are the mean±SD (n=3 assays per concentration). 5 ng of AChE was used in each reaction. C) In vitro inhibition of cholinesterase activity by 50 μM potato SAs in human protein homogenates from normal colonic tissue (n=3 donors) and Caco-2 colonic epithelial cells. Activity was normalized to the activity of the vehicle-treated sample. Values shown are the mean±SD of three replicate assays per homogenate. 5 ng each of recombinant human AChE and BChE were used for pure protein reactions, while 35 μg total protein was used for each homogenate. Multiplicity-adjusted P values were calculated using Tukey’s multiple comparison tests. D) Inhibition of recombinant human AChE activity by spent media from human stool (n=11 donors) incubated with 20 μM solanine in SAAC minimal medium. Activity was normalized to the activity of AChE treated with spent media from the same donor sample incubated with vehicle only. Values shown are the mean±SD of three AChE activity assays for each donor and compound combination. The relative abundances of select metabolites following incubation with solanine are shown below for each stool sample. Values shown are EIC peak areas normalized to the maximum peak area across all samples. P values were determined using a two-sided Welch’s T test. Ns: not significant with P>0.05.

Article Snippet: Steroidal alkaloid compounds were acquired from commercial sources: tomatine (Tokyo Chemical Industry), tomatidine (Cayman Chemical Company), solanine (Sigma Aldrich), chaconine (Sigma Aldrich), solanidine (Carbosynth), solamargine (MedChemExpress), solasonine (MedChemExpress), solasodine (Santa Cruz Biotechnology).

Techniques: Concentration Assay, In Vitro, Inhibition, Recombinant, Activity Assay, Comparison, Incubation

Journal: bioRxiv

Article Title: Gut microbiota gate host exposure to metabolites from dietary Solanums

doi: 10.1101/2024.03.20.584512

Figure Lengend Snippet: A) Tomatine and tomatidine concentrations in serum (nM), cecal contents (pmol/mg fresh weight), and liver (pmol/mg fresh weight) following a 10% tomato dietary intervention in C57BL/6J mice receiving antibiotics (Abx). Values shown are the mean±SD for each group, with individual replicates overlaid (n=5 mice per treatment). P values are determined using a two-sided Welch’s T test. ns = not significant (P > 0.05). B) Tomatine-derived metabolites in tissues following dietary intervention with 10% tomato diet in C57BL/6J mice receiving antibiotics. Abundances for each metabolite are normalized to the maximum EIC peak area across tissues shown in bars. Peak areas were normalized by sample dry mass to enable comparison between different tissue types. Values shown are the mean for each treatment group (n=5 mice per treatment). Structures as drawn are hypothesized based on m/z and MS/MS spectra; the position of hydroxylation is not known. For molecules with multiple detected isomers, the most abundant isomer is shown. C) Chaconine, solanine, and solanidine concentrations in serum (nM), cecal contents (pmol/mg fresh weight), and liver (pmol/mg fresh weight) following a 5% potato dietary intervention in C57BL/6J mice receiving antibiotics. Values shown are the mean±SD for each treatment group, with individual replicates overlaid (n=5 mice per treatment). P values are determined using a two-sided Welch’s T test. ns = not significant (P > 0.05). D) Solanidine and chaconine in human urine from colectomy patients (n=12 donors) and donors with intact colons (n=17 donors). Compound abundance is shown as EIC peak area normalized to the creatinine content in that sample. Values shown are for individual donor samples, with the molar concentration of solanidine in one intact colon sample shown in pmol solanidine per mg creatinine.

Article Snippet: Steroidal alkaloid compounds were acquired from commercial sources: tomatine (Tokyo Chemical Industry), tomatidine (Cayman Chemical Company), solanine (Sigma Aldrich), chaconine (Sigma Aldrich), solanidine (Carbosynth), solamargine (MedChemExpress), solasonine (MedChemExpress), solasodine (Santa Cruz Biotechnology).

Techniques: Derivative Assay, Comparison, Tandem Mass Spectroscopy, Concentration Assay

Journal: bioRxiv

Article Title: Gut microbiota gate host exposure to metabolites from dietary Solanums

doi: 10.1101/2024.03.20.584512

Figure Lengend Snippet: A) Total steroidal alkaloid concentration in the serum of C57BL/6J mice fed potato-supplemented diet and treated with antibiotics. Aglycone concentrations were estimated using a solanidine standard curve. Data shown are the mean±SD (n=5 mice). P values were determined using a two-sided Welch’s T test. B) In vitro inhibition of recombinant human AChE activity by potato steroidal alkaloids. Data shown are normalized to AChE treated with vehicle only (DMSO for solanine, chaconine and ethanol for solanidine) with a total vehicle concentration of 1% for all treatments. Values shown are the mean±SD (n=3 assays per concentration). 5 ng of AChE was used in each reaction. C) In vitro inhibition of cholinesterase activity by 50 μM potato SAs in human protein homogenates from normal colonic tissue (n=3 donors) and Caco-2 colonic epithelial cells. Activity was normalized to the activity of the vehicle-treated sample. Values shown are the mean±SD of three replicate assays per homogenate. 5 ng each of recombinant human AChE and BChE were used for pure protein reactions, while 35 μg total protein was used for each homogenate. Multiplicity-adjusted P values were calculated using Tukey’s multiple comparison tests. D) Inhibition of recombinant human AChE activity by spent media from human stool (n=11 donors) incubated with 20 μM solanine in SAAC minimal medium. Activity was normalized to the activity of AChE treated with spent media from the same donor sample incubated with vehicle only. Values shown are the mean±SD of three AChE activity assays for each donor and compound combination. The relative abundances of select metabolites following incubation with solanine are shown below for each stool sample. Values shown are EIC peak areas normalized to the maximum peak area across all samples. P values were determined using a two-sided Welch’s T test. Ns: not significant with P>0.05.

Article Snippet: Steroidal alkaloid compounds were acquired from commercial sources: tomatine (Tokyo Chemical Industry), tomatidine (Cayman Chemical Company), solanine (Sigma Aldrich), chaconine (Sigma Aldrich), solanidine (Carbosynth), solamargine (MedChemExpress), solasonine (MedChemExpress), solasodine (Santa Cruz Biotechnology).

Techniques: Concentration Assay, In Vitro, Inhibition, Recombinant, Activity Assay, Comparison, Incubation