Journal: Frontiers in Immunology

Article Title: High Content Image Analysis as a Tool to Morphologically Distinguish Macrophage Activation and Determine Its Importance for Foamy Alveolar Macrophage Responses

doi: 10.3389/fimmu.2021.611280

Figure Lengend Snippet: NR8383 rat macrophage activation status after stimulation with (A) LPS and IFN-γ, (B) IL-4 or (C) IL-4 or IL-4/IL-13. Data are normalized to cell count and presented as mean values ± SEM of three separate experiments. Statistical comparisons are made using a one-way ANOVA analysis with Bonferroni’s multiple comparison post-hoc test. Results are compared to non-polarized control (M0) and statistical significance is marked as follows: * indicates p < 0.01. (C) Representative fluorescent dot (quadrants) plots showing percentage of cells expressing MRC-1 surface receptor.

Article Snippet: Lipopolysaccharide (LPS, cat#L2630, Sigma Aldrich, Dorset, UK), recombinant rat interferon gamma (IFN-γ, cat#550072), and recombinant rat interleukin 4 (IL-4, cat#555107) were purchased from BD Biosciences, (Wokingham, Berkshire, UK), while recombinant rat interleukin 13 (IL-13, cat#CB243IL13) was purchased from PAN-Biotech (Wimborne, Dorset, UK).

Techniques: Activation Assay, Cell Counting, Expressing

Journal: Frontiers in Immunology

Article Title: High Content Image Analysis as a Tool to Morphologically Distinguish Macrophage Activation and Determine Its Importance for Foamy Alveolar Macrophage Responses

doi: 10.3389/fimmu.2021.611280

Figure Lengend Snippet: Morphological characteristic of non-activated, M(LPS+/IFN-γ+) or M(IL-4+) activated NR8383 rat macrophages. Nuclear area (A) , cellular area (B) , number vacuoles per cell (C) , average area of a single vacuole (D) , cellular area occupied by all vacuoles (E) and total area of all vacuoles per cell (F) were quantified in non-activated macrophages (M0), activated with 100 ng/mL LPS and 20 ng/mL IFN-γ [M(LPS+/IFN-γ+)] or activated with 30 ng/mL IL-4 [M(IL-4+)]. Data are presented as mean values ± SEM of three separate experiments. Statistical comparisons are made using a one-way ANOVA analysis with Bonferroni’s multiple comparison post-hoc test. Results are compared to non-polarized control (M0); *** indicates p < 0.001.

Article Snippet: Lipopolysaccharide (LPS, cat#L2630, Sigma Aldrich, Dorset, UK), recombinant rat interferon gamma (IFN-γ, cat#550072), and recombinant rat interleukin 4 (IL-4, cat#555107) were purchased from BD Biosciences, (Wokingham, Berkshire, UK), while recombinant rat interleukin 13 (IL-13, cat#CB243IL13) was purchased from PAN-Biotech (Wimborne, Dorset, UK).

Techniques:

Journal: Frontiers in Immunology

Article Title: High Content Image Analysis as a Tool to Morphologically Distinguish Macrophage Activation and Determine Its Importance for Foamy Alveolar Macrophage Responses

doi: 10.3389/fimmu.2021.611280

Figure Lengend Snippet: Functional characteristic of non-activated, M(LPS+/IFN-γ+) or M(IL-4+) activated NR8383 rat macrophages. Cell count (A) , phagocytic activity (B) , cellular phospholipid (C) and cellular neutral lipid content (D) were quantified in non-activated macrophages (M0), activated with 100 ng/ml LPS and 20 ng/ml IFN-γ [M(LPS+/IFN-γ+)] or activated with 30 ng/ml IL-4 [M(IL-4+)]. Data are normalized to cell count and presented as mean values ± SEM of three separate experiments. Statistical comparisons are made using a one-way ANOVA analysis with Bonferroni’s multiple comparison post-hoc test. Results are compared to non-polarized control (M0); ** indicate p < 0.01, *** indicate p < 0.001.

Article Snippet: Lipopolysaccharide (LPS, cat#L2630, Sigma Aldrich, Dorset, UK), recombinant rat interferon gamma (IFN-γ, cat#550072), and recombinant rat interleukin 4 (IL-4, cat#555107) were purchased from BD Biosciences, (Wokingham, Berkshire, UK), while recombinant rat interleukin 13 (IL-13, cat#CB243IL13) was purchased from PAN-Biotech (Wimborne, Dorset, UK).

Techniques: Functional Assay, Cell Counting, Activity Assay

Journal: Frontiers in Immunology

Article Title: High Content Image Analysis as a Tool to Morphologically Distinguish Macrophage Activation and Determine Its Importance for Foamy Alveolar Macrophage Responses

doi: 10.3389/fimmu.2021.611280

Figure Lengend Snippet: Effect on arginase activity and MRC-1 receptor expression in M0-non-activated and IL-4 activated NR8383 rat macrophages after 24 h exposure to 5 µM amiodarone and 0.1 µM staurosporine. (A) Effect on arginase activity. Data are normalized to cell count and presented as mean values ± SEM of three separate experiments. Statistical comparisons are made using a one-way ANOVA analysis with Bonferroni’s multiple comparison post-hoc test. Results are compared to non-polarized control (M0); *** indicates p < 0.001. (B) Representative fluorescent dot (quadrants) plots showing percentage of cells expressing MRC-1 surface receptor.

Article Snippet: Lipopolysaccharide (LPS, cat#L2630, Sigma Aldrich, Dorset, UK), recombinant rat interferon gamma (IFN-γ, cat#550072), and recombinant rat interleukin 4 (IL-4, cat#555107) were purchased from BD Biosciences, (Wokingham, Berkshire, UK), while recombinant rat interleukin 13 (IL-13, cat#CB243IL13) was purchased from PAN-Biotech (Wimborne, Dorset, UK).

Techniques: Activity Assay, Expressing, Cell Counting

Journal: Frontiers in Immunology

Article Title: High Content Image Analysis as a Tool to Morphologically Distinguish Macrophage Activation and Determine Its Importance for Foamy Alveolar Macrophage Responses

doi: 10.3389/fimmu.2021.611280

Figure Lengend Snippet: Phagocytic activity in M0-non-activated, LPS/IFN-γ or IL-4 activated NR8383 rat macrophages after 24 h exposure to 5 µM amiodarone (orange bars) and 0.1 µM staurosporine (purple bars). Data are expressed as a percentage of the phagocytic activity of non-polarized untreated NR8383 cells. Data represent the mean values ± SEM of three separate experiments. Statistical comparisons are made using a one-way ANOVA analysis with Bonferroni’s multiple comparison post-hoc test. Results are compared to non-polarized control (M0); ** indicate p < 0.01, *** indicates p < 0.001.

Article Snippet: Lipopolysaccharide (LPS, cat#L2630, Sigma Aldrich, Dorset, UK), recombinant rat interferon gamma (IFN-γ, cat#550072), and recombinant rat interleukin 4 (IL-4, cat#555107) were purchased from BD Biosciences, (Wokingham, Berkshire, UK), while recombinant rat interleukin 13 (IL-13, cat#CB243IL13) was purchased from PAN-Biotech (Wimborne, Dorset, UK).

Techniques: Activity Assay

Journal: Frontiers in Immunology

Article Title: High Content Image Analysis as a Tool to Morphologically Distinguish Macrophage Activation and Determine Its Importance for Foamy Alveolar Macrophage Responses

doi: 10.3389/fimmu.2021.611280

Figure Lengend Snippet: Phospholipid (A) and neutral lipid (B) content in M0-non-activated, LPS/IFN-γ and IL-4 activated NR8383 rat macrophages after 24 h exposure to 5 µM amiodarone (orange bars) and 0.1 µM staurosporine (purple bars). Data are normalized to cell count and presented as mean values ± SEM of three separate experiments. Statistical comparisons are made using a one-way ANOVA analysis with Bonferroni’s multiple comparison post-hoc test. Results are compared to non-polarized control (M0); *** indicates p < 0.001.

Article Snippet: Lipopolysaccharide (LPS, cat#L2630, Sigma Aldrich, Dorset, UK), recombinant rat interferon gamma (IFN-γ, cat#550072), and recombinant rat interleukin 4 (IL-4, cat#555107) were purchased from BD Biosciences, (Wokingham, Berkshire, UK), while recombinant rat interleukin 13 (IL-13, cat#CB243IL13) was purchased from PAN-Biotech (Wimborne, Dorset, UK).

Techniques: Cell Counting

Journal: Environmental Science and Pollution Research International

Article Title: Rutin protects against gamma-irradiation and malathion-induced oxidative stress and inflammation through regulation of mir-129-3p, mir-200C-3p, and mir-210 gene expressions in rats’ kidney

doi: 10.1007/s11356-023-27166-z

Figure Lengend Snippet: Inflammatory markers levels in the kidney tissues. C, control; rutin, rutin-treated animals; IRR, gamma-irradiated animals; MT, malathion-treated animals; IRR/MT, gamma-irradiation/malathion-treated animals; IRR/rutin, gamma-irradiation/rutin-treated animals; MT/rutin, malathion/rutin-treated animals; IRR/MT/rutin, gamma-irradiation/malathion/rutin-treated animals; IL-1β, interleukin-1 beta; IL-2, interleukin-2; IL-4, interleukin-4; IL-6, interleukin 6; TNF-α, tumor necrosis factor-alpha; NF-κB, nuclear factor–kappa B. The statistical significance to control, IRR, MT, and IRR/MT are denoted by a, b, c, and d, respectively, at p < 0.05. Statistical significance was analyzed by one-way ANOVA with Tukey post hoc multiple comparisons. These inflammatory markers are assessed by ELISA technique

Article Snippet: ELISA kits for rats from MyBioSource were used for the assessment of interleukin-1 beta (IL-1β, Cat.No:MBS825017), interleukin-2 (IL-2, Cat.No:MBS2701041), interleukin-4 (IL-4, Cat.No:MBS2883072), interleukin-6 (IL-6, Cat.No:MBS355410), tumor necrosis factor-alpha (TNF-α, Cat.No:MBS824824), and nuclear factor-kappa B (NF-κB, Cat.No:MBS268833) levels in the kidney tissues.

Techniques: Control, Irradiation, Enzyme-linked Immunosorbent Assay

Journal: Molecules

Article Title: Biotransformation of Timosaponin BII into Seven Characteristic Metabolites by the Gut Microbiota

doi: 10.3390/molecules26133861

Figure Lengend Snippet: The structure and mass spectra of timosaponin BII. ( A ) The structural formula of timosaponin BII and the Glipizide (internal standard (IS)). ( B ) Extracted ion chromatogram (EIC) spectra of timosaponin BII and the internal standard (Glipizide). ( C ) The high-resolution mass spectra of timosaponin BII acquired by LC/MS-Q-TOF.

Article Snippet: Timosaponin BII (CAS, 136656-07-0; Cat Number, IT0640), timosaponin AIII (CAS, 41059-79-4; Cat Number, IT0630), Sarsasapogenin (CAS, 126-19-2; Cat Number, SS8160), and Glipizide (CAS, 29094-61-9; Cat Number, SG8680) were purchased from Solarbio Life Sciences Co., Ltd. (Beijing, China).

Techniques: Liquid Chromatography with Mass Spectroscopy

Journal: Molecules

Article Title: Biotransformation of Timosaponin BII into Seven Characteristic Metabolites by the Gut Microbiota

doi: 10.3390/molecules26133861

Figure Lengend Snippet: Timosaponin BII could be metabolized in the gut microbiota. Intestinal bacteria mainly participated in the metabolism of timosaponin BII by comparison between untreated and heated intestinal contents ( A , B ). ( A ) The level of timosaponin BII decreased during incubation with rat intestinal bacteria after 0 h, 1 h, 2 h, 6 h, 12 h, and 24 h. ( B ) The metabolic characteristic of timosaponin BII by gut microbiota in more precise spans of time. The liver may not be the metabolic site of timosaponin BII ( C , D ). ( C ) The metabolic curve of timosaponin BII by rat liver homogenate. ( D ) The metabolic curve of timosaponin BII by rat microsome. Data are presented as mean ± SD, and two-tailed Student’s t test were used for analysis (** p < 0.01, *** p < 0.001).

Article Snippet: Timosaponin BII (CAS, 136656-07-0; Cat Number, IT0640), timosaponin AIII (CAS, 41059-79-4; Cat Number, IT0630), Sarsasapogenin (CAS, 126-19-2; Cat Number, SS8160), and Glipizide (CAS, 29094-61-9; Cat Number, SG8680) were purchased from Solarbio Life Sciences Co., Ltd. (Beijing, China).

Techniques: Incubation, Two Tailed Test

Journal: Molecules

Article Title: Biotransformation of Timosaponin BII into Seven Characteristic Metabolites by the Gut Microbiota

doi: 10.3390/molecules26133861

Figure Lengend Snippet: ( A ) The extracted ion chromatograms (EICs) showed that the levels of the possible metabolites M1–M3 increased with increasing time. ( B ) The MS/MS data of the timosaponin BII metabolites M1–M3. ( C ) Possible structures and mass spectrometric cleavage pathway of metabolites M1–M3. Among them, the structure framed by a rectangle was reported in the gut microbiota for the first time.

Article Snippet: Timosaponin BII (CAS, 136656-07-0; Cat Number, IT0640), timosaponin AIII (CAS, 41059-79-4; Cat Number, IT0630), Sarsasapogenin (CAS, 126-19-2; Cat Number, SS8160), and Glipizide (CAS, 29094-61-9; Cat Number, SG8680) were purchased from Solarbio Life Sciences Co., Ltd. (Beijing, China).

Techniques: Tandem Mass Spectroscopy

Journal: Molecules

Article Title: Biotransformation of Timosaponin BII into Seven Characteristic Metabolites by the Gut Microbiota

doi: 10.3390/molecules26133861

Figure Lengend Snippet: ( A ) The extracted ion chromatograms (EICs) showed that the levels of the possible metabolites M4 and M5 increased with increasing time. ( B ) The MS/MS data of the timosaponin BII metabolites M4 and M5. ( C ) Possible structures and mass spectrometric cleavage pathway of metabolites M4 and M5. Among them, the structure framed by a rectangle was reported in the gut microbiota for the first time.

Article Snippet: Timosaponin BII (CAS, 136656-07-0; Cat Number, IT0640), timosaponin AIII (CAS, 41059-79-4; Cat Number, IT0630), Sarsasapogenin (CAS, 126-19-2; Cat Number, SS8160), and Glipizide (CAS, 29094-61-9; Cat Number, SG8680) were purchased from Solarbio Life Sciences Co., Ltd. (Beijing, China).

Techniques: Tandem Mass Spectroscopy

Journal: Molecules

Article Title: Biotransformation of Timosaponin BII into Seven Characteristic Metabolites by the Gut Microbiota

doi: 10.3390/molecules26133861

Figure Lengend Snippet: ( A ) The extracted ion chromatograms (EICs) showed that the levels of the possible metabolites M6 and M7 increased with increasing time. ( B ) The MS/MS data of the timosaponin BII metabolites M6 and M7. ( C ) Possible structures and mass spectrometric cleavage pathway of metabolites M6 and M7. Among them, the structure framed by a rectangle was reported in the gut microbiota for the first time.

Article Snippet: Timosaponin BII (CAS, 136656-07-0; Cat Number, IT0640), timosaponin AIII (CAS, 41059-79-4; Cat Number, IT0630), Sarsasapogenin (CAS, 126-19-2; Cat Number, SS8160), and Glipizide (CAS, 29094-61-9; Cat Number, SG8680) were purchased from Solarbio Life Sciences Co., Ltd. (Beijing, China).

Techniques: Tandem Mass Spectroscopy

Journal: Molecules

Article Title: Biotransformation of Timosaponin BII into Seven Characteristic Metabolites by the Gut Microbiota

doi: 10.3390/molecules26133861

Figure Lengend Snippet: Characteristics of Timosaponin BII metabolites in gut microbiota by LC/MS-Q-TOF.

Article Snippet: Timosaponin BII (CAS, 136656-07-0; Cat Number, IT0640), timosaponin AIII (CAS, 41059-79-4; Cat Number, IT0630), Sarsasapogenin (CAS, 126-19-2; Cat Number, SS8160), and Glipizide (CAS, 29094-61-9; Cat Number, SG8680) were purchased from Solarbio Life Sciences Co., Ltd. (Beijing, China).

Techniques: Molecular Weight

Journal: Molecules

Article Title: Biotransformation of Timosaponin BII into Seven Characteristic Metabolites by the Gut Microbiota

doi: 10.3390/molecules26133861

Figure Lengend Snippet: The level of possible timosaponin BII metabolites changed during incubation with rat intestinal bacteria after 0 min, 15 min, 30 min, 60 min, 90 min, and 120 min. ( A ) Metabolic curve of timosaponin AIII. ( B ) Metabolic curve of timosaponin AI. ( C ) Metabolic curve of sarsasapogenin. ( D ) The temporal characteristics of potential timosaponin BII metabolites by gut microbiota. ( E ) Extracted ion chromatogram (EIC) spectra of timosaponin AIII, timosaponin AI, sarsasapogenin and the internal standard (Glipizide). ( F ) Metabolic pathway of timosaponin BII and possible structures of metabolites.

Article Snippet: Timosaponin BII (CAS, 136656-07-0; Cat Number, IT0640), timosaponin AIII (CAS, 41059-79-4; Cat Number, IT0630), Sarsasapogenin (CAS, 126-19-2; Cat Number, SS8160), and Glipizide (CAS, 29094-61-9; Cat Number, SG8680) were purchased from Solarbio Life Sciences Co., Ltd. (Beijing, China).

Techniques: Incubation