Structured Review

SCIEX tbii
<t>TAIII</t> is the selectively cytotoxic compound in BN108 extract. A. Three cell lines indicated were treated with a range of concentrations of TAIII for 24 hours, and cell viability was analyzed by Annexin-PI staining. B. Structures of TAIII, <t>TBII</t> and sarsasapogenin. C. Cells lines were treated for 24 hours with TBII at 40 µM, heat–inactivated laminarinase alone (0.5 µg/ml), or a mixture of both after incubation for 30 min at 50°C and heat-inactivation of the enzyme. MS analysis indicated that about 20 to 30% of TBIII was converted to TAIII. Apoptotic cells were identified by Annexin V binding. The results are representative of the two separate experiments performed with very similar results.
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1) Product Images from "Timosaponin AIII Is Preferentially Cytotoxic to Tumor Cells through Inhibition of mTOR and Induction of ER Stress"

Article Title: Timosaponin AIII Is Preferentially Cytotoxic to Tumor Cells through Inhibition of mTOR and Induction of ER Stress

Journal: PLoS ONE

doi: 10.1371/journal.pone.0007283

TAIII is the selectively cytotoxic compound in BN108 extract. A. Three cell lines indicated were treated with a range of concentrations of TAIII for 24 hours, and cell viability was analyzed by Annexin-PI staining. B. Structures of TAIII, TBII and sarsasapogenin. C. Cells lines were treated for 24 hours with TBII at 40 µM, heat–inactivated laminarinase alone (0.5 µg/ml), or a mixture of both after incubation for 30 min at 50°C and heat-inactivation of the enzyme. MS analysis indicated that about 20 to 30% of TBIII was converted to TAIII. Apoptotic cells were identified by Annexin V binding. The results are representative of the two separate experiments performed with very similar results.
Figure Legend Snippet: TAIII is the selectively cytotoxic compound in BN108 extract. A. Three cell lines indicated were treated with a range of concentrations of TAIII for 24 hours, and cell viability was analyzed by Annexin-PI staining. B. Structures of TAIII, TBII and sarsasapogenin. C. Cells lines were treated for 24 hours with TBII at 40 µM, heat–inactivated laminarinase alone (0.5 µg/ml), or a mixture of both after incubation for 30 min at 50°C and heat-inactivation of the enzyme. MS analysis indicated that about 20 to 30% of TBIII was converted to TAIII. Apoptotic cells were identified by Annexin V binding. The results are representative of the two separate experiments performed with very similar results.

Techniques Used: Staining, Incubation, Mass Spectrometry, Binding Assay

2) Product Images from "Characterizing the Developmental Trajectory of Sirolimus Clearance in Neonates and Infants"

Article Title: Characterizing the Developmental Trajectory of Sirolimus Clearance in Neonates and Infants

Journal: CPT: Pharmacometrics & Systems Pharmacology

doi: 10.1002/psp4.12096

The developmental trajectory of sirolimus clearance over age. A maximum a posteriori probability (MAP) Bayesian sirolimus clearance (CL) estimate was generated using sirolimus concentration measurement at each sampling point. Open circles represent observed sirolimus CL estimates size‐normalized by an allometric scaling with a power exponent of 0.67 ( n = 316 points from 24 patients younger than 4 years old). The patient demographics for this analysis are summarized in Supplemental Table 2 . Seven patients were on weaning doses of steroids as part of the initial treatment in kaposiform hemangioendothelioma (KHE) patients and the doses were tapered off after 1 to 2 months. As steroid administration in KHE patients did not show a significant effect on the estimation of population parameters in the covariate analysis, their data were included in the analysis. A black line represents the developmental trajectory of sirolimus clearance based on mean population parameter estimates of the sigmoidal E max model: CL matured (CL at fully matured level), 18.1 L/h/70kg; TM 50 (postmenstrual age at which CL is half of CL matured ), 62.9; and Hill coefficient, 2.94.
Figure Legend Snippet: The developmental trajectory of sirolimus clearance over age. A maximum a posteriori probability (MAP) Bayesian sirolimus clearance (CL) estimate was generated using sirolimus concentration measurement at each sampling point. Open circles represent observed sirolimus CL estimates size‐normalized by an allometric scaling with a power exponent of 0.67 ( n = 316 points from 24 patients younger than 4 years old). The patient demographics for this analysis are summarized in Supplemental Table 2 . Seven patients were on weaning doses of steroids as part of the initial treatment in kaposiform hemangioendothelioma (KHE) patients and the doses were tapered off after 1 to 2 months. As steroid administration in KHE patients did not show a significant effect on the estimation of population parameters in the covariate analysis, their data were included in the analysis. A black line represents the developmental trajectory of sirolimus clearance based on mean population parameter estimates of the sigmoidal E max model: CL matured (CL at fully matured level), 18.1 L/h/70kg; TM 50 (postmenstrual age at which CL is half of CL matured ), 62.9; and Hill coefficient, 2.94.

Techniques Used: Generated, Concentration Assay, Sampling

Age‐dependent change in the metabolite formation of sirolimus. The ratios of metabolite concentration to sirolimus concentration were described against age by a simple E max model for 25‐hydroxysirolimus (25‐OH, A) and 16‐O‐demethysirolimus (16‐O‐DM, B). Solid lines show the regression lines. Dashed lines show 95% confidence intervals of prediction band.
Figure Legend Snippet: Age‐dependent change in the metabolite formation of sirolimus. The ratios of metabolite concentration to sirolimus concentration were described against age by a simple E max model for 25‐hydroxysirolimus (25‐OH, A) and 16‐O‐demethysirolimus (16‐O‐DM, B). Solid lines show the regression lines. Dashed lines show 95% confidence intervals of prediction band.

Techniques Used: Concentration Assay

Visual predictive check of the NONMEM model. Open circles, observed allometrically size‐normalized sirolimus clearance; solid red line, median of observed; dashed red lines, lower (5th) and upper (95th) percentiles of the observed data; solid black line, median of predicted data; dashed black lines, 5th and 95th percentiles of the predicted data; shaded areas, confidence intervals around the prediction intervals in each bin.
Figure Legend Snippet: Visual predictive check of the NONMEM model. Open circles, observed allometrically size‐normalized sirolimus clearance; solid red line, median of observed; dashed red lines, lower (5th) and upper (95th) percentiles of the observed data; solid black line, median of predicted data; dashed black lines, 5th and 95th percentiles of the predicted data; shaded areas, confidence intervals around the prediction intervals in each bin.

Techniques Used:

3) Product Images from "Progression of pathology in PINK1-deficient mouse brain from splicing via ubiquitination, ER stress, and mitophagy changes to neuroinflammation"

Article Title: Progression of pathology in PINK1-deficient mouse brain from splicing via ubiquitination, ER stress, and mitophagy changes to neuroinflammation

Journal: Journal of Neuroinflammation

doi: 10.1186/s12974-017-0928-0

Astroglial and microglial response, as well as ceramide accumulation in Pink1 -deficient mouse brain. The immunohistochemical staining of GFAP as astroglial marker and Iba1 as microglial marker in Pink1 − /− mouse brain at the age of 18 months demonstrated a mild increase in the frequency of inflammatory glial reactions, best detectable along the corticospinal tract where it traverses the brainstem ( a , 10× objective used) and the striatal region ( b , 4× objective). c The elevated frequency of this glial response was demonstrated in cell counts for astrogliosis and microgliosis in relation to total cells per area (nuclear counterstain) in 18-month-old brains that underwent automated immunohistochemistry of GFAP1 and Iba1 demonstrated region-specific mild effects. The substantia nigra was less affected than areas that are penetrated by myelinated axons. Data are displayed as bar graphs , illustrating mean values of serial sectioned and randomly picked mouse brain areas. Brain graphs show standard error of the mean (SEM) of technical replicates with at least n = 3 sections per genotype per region. Statistical analysis would be inappropriate, given that the serial sections were from too few inbred animals. d Increased levels of ceramides, glucosyl-ceramides, and lactosyl-ceramides were observed by liquid chromatography tandem mass spectrometry in the olfactory bulb summarized for ages of 9–13 months, 17 months, and 21 months ( n = 3 or 4 mice of each age and genotype). The scatter shows individual mice, the line is the mean, and whiskers show the standard deviation. e Log2-transformed total ceramides across brain regions reveal age-dependent differences between genotype, i.e., an increase over time in wildtype mice that contrast with very high levels in younger Pink1 −/− mice which remain at this elevated level. Two-way ANOVA differed significantly between genotypes. Asterisks illustrate the significance (multivariate ANOVA, subsequent t test for each lipid, Holm-Šidák adjusted p values, * p
Figure Legend Snippet: Astroglial and microglial response, as well as ceramide accumulation in Pink1 -deficient mouse brain. The immunohistochemical staining of GFAP as astroglial marker and Iba1 as microglial marker in Pink1 − /− mouse brain at the age of 18 months demonstrated a mild increase in the frequency of inflammatory glial reactions, best detectable along the corticospinal tract where it traverses the brainstem ( a , 10× objective used) and the striatal region ( b , 4× objective). c The elevated frequency of this glial response was demonstrated in cell counts for astrogliosis and microgliosis in relation to total cells per area (nuclear counterstain) in 18-month-old brains that underwent automated immunohistochemistry of GFAP1 and Iba1 demonstrated region-specific mild effects. The substantia nigra was less affected than areas that are penetrated by myelinated axons. Data are displayed as bar graphs , illustrating mean values of serial sectioned and randomly picked mouse brain areas. Brain graphs show standard error of the mean (SEM) of technical replicates with at least n = 3 sections per genotype per region. Statistical analysis would be inappropriate, given that the serial sections were from too few inbred animals. d Increased levels of ceramides, glucosyl-ceramides, and lactosyl-ceramides were observed by liquid chromatography tandem mass spectrometry in the olfactory bulb summarized for ages of 9–13 months, 17 months, and 21 months ( n = 3 or 4 mice of each age and genotype). The scatter shows individual mice, the line is the mean, and whiskers show the standard deviation. e Log2-transformed total ceramides across brain regions reveal age-dependent differences between genotype, i.e., an increase over time in wildtype mice that contrast with very high levels in younger Pink1 −/− mice which remain at this elevated level. Two-way ANOVA differed significantly between genotypes. Asterisks illustrate the significance (multivariate ANOVA, subsequent t test for each lipid, Holm-Šidák adjusted p values, * p

Techniques Used: Immunohistochemistry, Staining, Marker, Liquid Chromatography, Mass Spectrometry, Mouse Assay, Standard Deviation, Transformation Assay

4) Product Images from "Orally active microtubule-targeting agent, MPT0B271, for the treatment of human non-small cell lung cancer, alone and in combination with erlotinib"

Article Title: Orally active microtubule-targeting agent, MPT0B271, for the treatment of human non-small cell lung cancer, alone and in combination with erlotinib

Journal: Cell Death & Disease

doi: 10.1038/cddis.2014.128

( a ) Measurement of apoptosis. A549 cells were treated with the indicated concentration of MPT0B271, and oligonucleosomal DNA fragmentation was quantitatively assessed with the Cell Death ELISA PLUS kit. Apoptosis was enhanced in relation to control cells. Data are expressed as the mean±S.E. of at least three independent experiments. *** P
Figure Legend Snippet: ( a ) Measurement of apoptosis. A549 cells were treated with the indicated concentration of MPT0B271, and oligonucleosomal DNA fragmentation was quantitatively assessed with the Cell Death ELISA PLUS kit. Apoptosis was enhanced in relation to control cells. Data are expressed as the mean±S.E. of at least three independent experiments. *** P

Techniques Used: Concentration Assay, Enzyme-linked Immunosorbent Assay

( a ) The effect of MPT0B271 on p53 expression and phosphorylation in A549 cells. Cells were treated with MPT0B271 (0.3 μ M) for the indicated times, and whole-cell extracts were prepared and underwent western blot analysis using the indicated antibodies. ( b ) A549 cells were pretreated with or without p53 siRNA for 24 h and then incubated with or without MPT0B271 (0.1 μ M) for 48 h. Upper panel , total cellular lysates were subjected to western blot analysis of p53 and PARP. Lower panel , cell viability was measured by the SRB assay and expressed as a percentage of the untreated control. ( c ) Concentration-dependent effect of MPT0B271 on cell viability. H1299 and H226 cells were treated with or without the indicated concentration of MPT0B271 for 48 h, and the cytotoxic effect was determined with the MTT assay. Data are expressed as the mean±S.E. of at least three independent experiments. * P
Figure Legend Snippet: ( a ) The effect of MPT0B271 on p53 expression and phosphorylation in A549 cells. Cells were treated with MPT0B271 (0.3 μ M) for the indicated times, and whole-cell extracts were prepared and underwent western blot analysis using the indicated antibodies. ( b ) A549 cells were pretreated with or without p53 siRNA for 24 h and then incubated with or without MPT0B271 (0.1 μ M) for 48 h. Upper panel , total cellular lysates were subjected to western blot analysis of p53 and PARP. Lower panel , cell viability was measured by the SRB assay and expressed as a percentage of the untreated control. ( c ) Concentration-dependent effect of MPT0B271 on cell viability. H1299 and H226 cells were treated with or without the indicated concentration of MPT0B271 for 48 h, and the cytotoxic effect was determined with the MTT assay. Data are expressed as the mean±S.E. of at least three independent experiments. * P

Techniques Used: Expressing, Western Blot, Incubation, Sulforhodamine B Assay, Concentration Assay, MTT Assay

( a ) Various types of human cancer cells were treated with the indicated concentrations of MPT0B271 for 48 h. Then, cell growth inhibition was determined using the SRB assay, and the GI 50 of each cell line is expressed as the mean±S.E. of four independent determinations. ( b ) The cytotoxic effects of various human cancer cell lines were determined using an MTT assay. The IC 50 of each cell line is expressed as the mean±S.E. of four independent determinations. ( c ) NCI/ADR-RES cells were treated with the indicated concentration of paclitaxel or vincristine for 48 h, and cell growth was determined by the SRB assay. Data are expressed as the mean±S.E. of at least three independent experiments. *** P
Figure Legend Snippet: ( a ) Various types of human cancer cells were treated with the indicated concentrations of MPT0B271 for 48 h. Then, cell growth inhibition was determined using the SRB assay, and the GI 50 of each cell line is expressed as the mean±S.E. of four independent determinations. ( b ) The cytotoxic effects of various human cancer cell lines were determined using an MTT assay. The IC 50 of each cell line is expressed as the mean±S.E. of four independent determinations. ( c ) NCI/ADR-RES cells were treated with the indicated concentration of paclitaxel or vincristine for 48 h, and cell growth was determined by the SRB assay. Data are expressed as the mean±S.E. of at least three independent experiments. *** P

Techniques Used: Inhibition, Sulforhodamine B Assay, MTT Assay, Concentration Assay

( a ) Effect of MPT0B271 on cell-cycle progression. A549 cells were exposed to 0.3 μ M MPT0B271 for the indicated times and then stained with PI to determine the proportion of DNA. Data acquisition and analysis were performed on a FACScan flow cytometer. The data are expressed as the mean±S.E. of at least three independent experiments. ( b ) The effect of MPT0B271 on G2/M cell-cycle regulatory proteins. A549 cells were treated with 0.3 μ M MPT0B271 for the indicated times. Whole-cell extracts were subjected to SDS-PAGE and immunoblotted with the indicated antibodies. ( c ) Treatment with 0.3 μ M MPT0B271 for the indicated times. Nuclear lysates were subjected to western blot analysis using an antibody specific for cyclin B1
Figure Legend Snippet: ( a ) Effect of MPT0B271 on cell-cycle progression. A549 cells were exposed to 0.3 μ M MPT0B271 for the indicated times and then stained with PI to determine the proportion of DNA. Data acquisition and analysis were performed on a FACScan flow cytometer. The data are expressed as the mean±S.E. of at least three independent experiments. ( b ) The effect of MPT0B271 on G2/M cell-cycle regulatory proteins. A549 cells were treated with 0.3 μ M MPT0B271 for the indicated times. Whole-cell extracts were subjected to SDS-PAGE and immunoblotted with the indicated antibodies. ( c ) Treatment with 0.3 μ M MPT0B271 for the indicated times. Nuclear lysates were subjected to western blot analysis using an antibody specific for cyclin B1

Techniques Used: Staining, Flow Cytometry, Cytometry, SDS Page, Western Blot

( a ) Chemical structure of MPT0B271. ( b ) Effect of MPT0B271 on tubulin polymerization. Purified tubulin in reaction buffer was incubated at 37°C in the absence or presence of increasing concentrations of MPT0B271, 10 μ M paclitaxel, 10 μ M colchicine, or 10 μ M vincristine. Assembly of microtubules was then measured over 30 min at 1 min intervals at an absorbance of 340 nm using a spectrophotometer. ( c ) Immunofluorescence staining of microtubules in A549 cells. Cells were treated with vehicle (DMSO), 0.3 μ M MPT0B271, paclitaxel, or vincristine for 24 h. Cells were labeled with a β -tubulin antibody and an FITC-conjugated anti-mouse IgG antibody, were counterstained with 4,6-diamidino-2-phenylindole (DAPI) and observed by confocal microscopy. Left , DAPI; middle , microtubule network; right , merged microtubule network and DAPI. ( d ) PK properties, plasma concentration versus time profiles of MPT0B271 after i.v. (2 mg/kg) and p.o. (20 mg/kg) dosing of fasted male CD-1 ( Crl. ) mice. ( e ) Efficacy of MPT0B271, dosed orally, on tumor xenografts. Upper panel , tumor growth of A549 xenografts in nude mice that were orally treated with or without MPT0B271 (5, 10, and 20 mg/kg). Tumor growth is presented as the mean tumor volume (mm 3 )±S.E. Tumor volume was determined using caliper measurements and was calculated as the product of 1/2 × length × width 2 . Lower panel , body weight (g) of the mice. * P
Figure Legend Snippet: ( a ) Chemical structure of MPT0B271. ( b ) Effect of MPT0B271 on tubulin polymerization. Purified tubulin in reaction buffer was incubated at 37°C in the absence or presence of increasing concentrations of MPT0B271, 10 μ M paclitaxel, 10 μ M colchicine, or 10 μ M vincristine. Assembly of microtubules was then measured over 30 min at 1 min intervals at an absorbance of 340 nm using a spectrophotometer. ( c ) Immunofluorescence staining of microtubules in A549 cells. Cells were treated with vehicle (DMSO), 0.3 μ M MPT0B271, paclitaxel, or vincristine for 24 h. Cells were labeled with a β -tubulin antibody and an FITC-conjugated anti-mouse IgG antibody, were counterstained with 4,6-diamidino-2-phenylindole (DAPI) and observed by confocal microscopy. Left , DAPI; middle , microtubule network; right , merged microtubule network and DAPI. ( d ) PK properties, plasma concentration versus time profiles of MPT0B271 after i.v. (2 mg/kg) and p.o. (20 mg/kg) dosing of fasted male CD-1 ( Crl. ) mice. ( e ) Efficacy of MPT0B271, dosed orally, on tumor xenografts. Upper panel , tumor growth of A549 xenografts in nude mice that were orally treated with or without MPT0B271 (5, 10, and 20 mg/kg). Tumor growth is presented as the mean tumor volume (mm 3 )±S.E. Tumor volume was determined using caliper measurements and was calculated as the product of 1/2 × length × width 2 . Lower panel , body weight (g) of the mice. * P

Techniques Used: Purification, Incubation, Spectrophotometry, Immunofluorescence, Staining, Labeling, Confocal Microscopy, Concentration Assay, Mouse Assay

In vitro and in vivo antitumor activity of MPT0B271 in combination with erlotinib. ( a ) A549 cells were treated with erlotinib (5 μ M) in combination with MPT0B271 (0.0125 or 0.025 μ M) for 48 h, and cell apoptosis was measured using the Cell Death ELISA PLUS kit. Data are expressed as the mean±S.E. of at least three independent determinations. ( b ) A549 xenograft model. A549-tumor-bearing nude mice were treated with vehicle, MPT0B271 (20 mg/kg/day by oral gavage q2d), erlotinib (25 mg/kg/day by oral gavage once a day), or MPT0B271 in combination with erlotinib. Tumor was excised when the tumor size reached 1200 mm 3 . ( c ) The body weight of the mice measured daily during the first week and then at the days of administration. * P
Figure Legend Snippet: In vitro and in vivo antitumor activity of MPT0B271 in combination with erlotinib. ( a ) A549 cells were treated with erlotinib (5 μ M) in combination with MPT0B271 (0.0125 or 0.025 μ M) for 48 h, and cell apoptosis was measured using the Cell Death ELISA PLUS kit. Data are expressed as the mean±S.E. of at least three independent determinations. ( b ) A549 xenograft model. A549-tumor-bearing nude mice were treated with vehicle, MPT0B271 (20 mg/kg/day by oral gavage q2d), erlotinib (25 mg/kg/day by oral gavage once a day), or MPT0B271 in combination with erlotinib. Tumor was excised when the tumor size reached 1200 mm 3 . ( c ) The body weight of the mice measured daily during the first week and then at the days of administration. * P

Techniques Used: In Vitro, In Vivo, Activity Assay, Enzyme-linked Immunosorbent Assay, Mouse Assay

5) Product Images from "Evaluating the Suitability of Using Rat Models for Preclinical Efficacy and Side Effects with Inhaled Corticosteroids Nanosuspension Formulations"

Article Title: Evaluating the Suitability of Using Rat Models for Preclinical Efficacy and Side Effects with Inhaled Corticosteroids Nanosuspension Formulations

Journal: Nanoscale Research Letters

doi: 10.1007/s11671-010-9597-y

Ciclesonide nano particles
Figure Legend Snippet: Ciclesonide nano particles

Techniques Used:

6) Product Images from "Timosaponin AIII Is Preferentially Cytotoxic to Tumor Cells through Inhibition of mTOR and Induction of ER Stress"

Article Title: Timosaponin AIII Is Preferentially Cytotoxic to Tumor Cells through Inhibition of mTOR and Induction of ER Stress

Journal: PLoS ONE

doi: 10.1371/journal.pone.0007283

Aqueous extract from Anemarrhena asphodels induces apoptosis selectively in cancer cells. A. The indicated cell lines (three breast carcinoma lines; three prostate carcinoma lines, three immortalized mammary epithelial lines) and primary fibroblasts IMR90 were treated with 0.5 mg/ml of BN108 for 24 hours. Cells were analyzed for apoptosis by flow cytometry of Annexin V/PI stained cells. Percentage of Annexin V- binding cells is shown; results are an average of at least three experiments. B. Cell cycle analysis of BT474 and MCF10A cells treated with 0.5 mg/ml BN108 for 48 hours. Numbers show percentages of cells with DNA content less than 2n. C. Western blot analysis of PARP cleavage in BT474 and normal MCF10A cells treated with BN108 for 24 hours. D. BT474 and IMR90 cells were treated for 1 or 2 days with BN108, fixed, and cells with activated caspases were identified by flow cytometry with the Vibrant FAM caspase assay kit (InVitrogen). E. Apoptotic cells were detected in BT474 cultures as described in 1A. Cells were treated for 24 hours with BN108 (0.5 mg/ml) or TAIII (2.5 µM) with or without caspase 4 inhibitor LEVD-fmk or caspase 3 inhibitor DEVD-fmk at 10 µM.
Figure Legend Snippet: Aqueous extract from Anemarrhena asphodels induces apoptosis selectively in cancer cells. A. The indicated cell lines (three breast carcinoma lines; three prostate carcinoma lines, three immortalized mammary epithelial lines) and primary fibroblasts IMR90 were treated with 0.5 mg/ml of BN108 for 24 hours. Cells were analyzed for apoptosis by flow cytometry of Annexin V/PI stained cells. Percentage of Annexin V- binding cells is shown; results are an average of at least three experiments. B. Cell cycle analysis of BT474 and MCF10A cells treated with 0.5 mg/ml BN108 for 48 hours. Numbers show percentages of cells with DNA content less than 2n. C. Western blot analysis of PARP cleavage in BT474 and normal MCF10A cells treated with BN108 for 24 hours. D. BT474 and IMR90 cells were treated for 1 or 2 days with BN108, fixed, and cells with activated caspases were identified by flow cytometry with the Vibrant FAM caspase assay kit (InVitrogen). E. Apoptotic cells were detected in BT474 cultures as described in 1A. Cells were treated for 24 hours with BN108 (0.5 mg/ml) or TAIII (2.5 µM) with or without caspase 4 inhibitor LEVD-fmk or caspase 3 inhibitor DEVD-fmk at 10 µM.

Techniques Used: Flow Cytometry, Cytometry, Staining, Binding Assay, Cell Cycle Assay, Western Blot, Caspase Assay

TAIII and BN108 induce similar changes in gene expression and inhibit major proliferative signal transduction pathways selectively in cancer cells. A. Both BN108 and TAIII induce expression of REDD1 and inhibit MYC in BT474 but not in MCF10A cells. Top panels, cell extracts were prepared fro untreated cells and cells treated with BN108 (0.5 mg/ml) and TAIII (2.5 µM in BT474 and 7.5 µM for MCF10A) for 4 hours. Bottom panels, time course of expression of the indicated proteins in BT474 and MCF10A cells treated with TAIII. B. Top, both BN108 and TAIII inhibit phosphorylation of mTORC1 targets s6 ribosomal protein and 4eBP1 as well as AKT kinase selectively in cancer cells. Bottom, time course of inhibition of activating posphorylations of AKT and mTOR, as well as inhibition of phosphorylation of mTORC1 targets s6 and 4eBP1by TAIII in BT474 cells.
Figure Legend Snippet: TAIII and BN108 induce similar changes in gene expression and inhibit major proliferative signal transduction pathways selectively in cancer cells. A. Both BN108 and TAIII induce expression of REDD1 and inhibit MYC in BT474 but not in MCF10A cells. Top panels, cell extracts were prepared fro untreated cells and cells treated with BN108 (0.5 mg/ml) and TAIII (2.5 µM in BT474 and 7.5 µM for MCF10A) for 4 hours. Bottom panels, time course of expression of the indicated proteins in BT474 and MCF10A cells treated with TAIII. B. Top, both BN108 and TAIII inhibit phosphorylation of mTORC1 targets s6 ribosomal protein and 4eBP1 as well as AKT kinase selectively in cancer cells. Bottom, time course of inhibition of activating posphorylations of AKT and mTOR, as well as inhibition of phosphorylation of mTORC1 targets s6 and 4eBP1by TAIII in BT474 cells.

Techniques Used: Expressing, Transduction, Inhibition

TAIII induces ER stress and protective autophagy. A. Treatment with BN108 induce phosphorylation of translation initiation factor eIF2α. Cell extracts were prepared from BT474 cells treated with BN108 (0.5 mg/ml) for times indicated and subjected to gel electrophoresis and blotting with antibodies against phosphorylated form of eIF2α (Ser51) or total eIF2α protein. B. Western blot analysis of the expression of some markers of ER stress in MDAMB231 and BT474 cells. Cells were treated with TAIII for the indicated times and after electrophoresis extract were immunobloted with indicated antibodies. C. Western blot analysis of MDAMB231 cells transfected with control or GRP78 expressing vector. D. Expression of exogenous GRP78 in MDAMB231 cells offers partial protection against apoptosis induced by TAIII (5 µM) or MG132 (0.25 µM).
Figure Legend Snippet: TAIII induces ER stress and protective autophagy. A. Treatment with BN108 induce phosphorylation of translation initiation factor eIF2α. Cell extracts were prepared from BT474 cells treated with BN108 (0.5 mg/ml) for times indicated and subjected to gel electrophoresis and blotting with antibodies against phosphorylated form of eIF2α (Ser51) or total eIF2α protein. B. Western blot analysis of the expression of some markers of ER stress in MDAMB231 and BT474 cells. Cells were treated with TAIII for the indicated times and after electrophoresis extract were immunobloted with indicated antibodies. C. Western blot analysis of MDAMB231 cells transfected with control or GRP78 expressing vector. D. Expression of exogenous GRP78 in MDAMB231 cells offers partial protection against apoptosis induced by TAIII (5 µM) or MG132 (0.25 µM).

Techniques Used: Nucleic Acid Electrophoresis, Western Blot, Expressing, Electrophoresis, Transfection, Plasmid Preparation

Autophagy induced by TAIII plays a protective role in cell death. A. Inhibition of autophagy augments apoptosis induced by TAIII in MDAMB231 and MCF10A cell lines, but not in BT474. Cells were pretreated for three hours with 20 µM of chloroquine (CQ) after which TAIII (4 µM for BT474, 5 µM for MM231 and 7.5 µM for MCF10A) was added for 24 hours. B. A cartoon illustrating the various cellular effects of BN108 and TAIII. The pathways induced selectively in cancer cells are shown in thicker lines.
Figure Legend Snippet: Autophagy induced by TAIII plays a protective role in cell death. A. Inhibition of autophagy augments apoptosis induced by TAIII in MDAMB231 and MCF10A cell lines, but not in BT474. Cells were pretreated for three hours with 20 µM of chloroquine (CQ) after which TAIII (4 µM for BT474, 5 µM for MM231 and 7.5 µM for MCF10A) was added for 24 hours. B. A cartoon illustrating the various cellular effects of BN108 and TAIII. The pathways induced selectively in cancer cells are shown in thicker lines.

Techniques Used: Inhibition

TAIII is the selectively cytotoxic compound in BN108 extract. A. Three cell lines indicated were treated with a range of concentrations of TAIII for 24 hours, and cell viability was analyzed by Annexin-PI staining. B. Structures of TAIII, TBII and sarsasapogenin. C. Cells lines were treated for 24 hours with TBII at 40 µM, heat–inactivated laminarinase alone (0.5 µg/ml), or a mixture of both after incubation for 30 min at 50°C and heat-inactivation of the enzyme. MS analysis indicated that about 20 to 30% of TBIII was converted to TAIII. Apoptotic cells were identified by Annexin V binding. The results are representative of the two separate experiments performed with very similar results.
Figure Legend Snippet: TAIII is the selectively cytotoxic compound in BN108 extract. A. Three cell lines indicated were treated with a range of concentrations of TAIII for 24 hours, and cell viability was analyzed by Annexin-PI staining. B. Structures of TAIII, TBII and sarsasapogenin. C. Cells lines were treated for 24 hours with TBII at 40 µM, heat–inactivated laminarinase alone (0.5 µg/ml), or a mixture of both after incubation for 30 min at 50°C and heat-inactivation of the enzyme. MS analysis indicated that about 20 to 30% of TBIII was converted to TAIII. Apoptotic cells were identified by Annexin V binding. The results are representative of the two separate experiments performed with very similar results.

Techniques Used: Staining, Incubation, Mass Spectrometry, Binding Assay

7) Product Images from "Evaluating the Suitability of Using Rat Models for Preclinical Efficacy and Side Effects with Inhaled Corticosteroids Nanosuspension Formulations"

Article Title: Evaluating the Suitability of Using Rat Models for Preclinical Efficacy and Side Effects with Inhaled Corticosteroids Nanosuspension Formulations

Journal: Nanoscale Research Letters

doi: 10.1007/s11671-010-9597-y

Fluticasone nano particles
Figure Legend Snippet: Fluticasone nano particles

Techniques Used:

8) Product Images from "Reduction of Ischemia/Reperfusion Injury With Bendavia, a Mitochondria-Targeting Cytoprotective Peptide"

Article Title: Reduction of Ischemia/Reperfusion Injury With Bendavia, a Mitochondria-Targeting Cytoprotective Peptide

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

doi: 10.1161/JAHA.112.001644

Uptake of Bendavia by the myocardium before ischemia (Baseline) and during reperfusion. Data represent the percentage of Bendavia taken up by the heart over a 1-minute time course.
Figure Legend Snippet: Uptake of Bendavia by the myocardium before ischemia (Baseline) and during reperfusion. Data represent the percentage of Bendavia taken up by the heart over a 1-minute time course.

Techniques Used:

Mitochondrial membrane potential (ΔΨ m ) in myocytes during cellular hypoxia/reoxygenation. A, Representative fluorescence images of myocytes loaded with the ΔΨ m sensor TMRM. ΔΨ m collapse often preceded cell death, and treatment with Bendavia improved the capacity to maintain ΔΨ m . B, Representative fluorescence intensity traces for cells in the study.
Figure Legend Snippet: Mitochondrial membrane potential (ΔΨ m ) in myocytes during cellular hypoxia/reoxygenation. A, Representative fluorescence images of myocytes loaded with the ΔΨ m sensor TMRM. ΔΨ m collapse often preceded cell death, and treatment with Bendavia improved the capacity to maintain ΔΨ m . B, Representative fluorescence intensity traces for cells in the study.

Techniques Used: Fluorescence

Left, Ischemic risk zone (fraction LV) and infarct size (fraction risk zone) in rabbits with risk zones > 20% of the LV exposed to in vivo ischemia/reperfusion (30 min/3 h) in combined Bendavia treatment group and vehicle group (mean±SEM; P =0.09, t test). Note similarity to data observed in the sheep model (text, Figure 1 ). Right, Relationship between the extent of the ischemic risk zone (g) and the extent of necrosis (g) in combined Bendavia group and vehicle group. Regression lines: vehicle is indicated by dashed line; Bendavia, solid line.
Figure Legend Snippet: Left, Ischemic risk zone (fraction LV) and infarct size (fraction risk zone) in rabbits with risk zones > 20% of the LV exposed to in vivo ischemia/reperfusion (30 min/3 h) in combined Bendavia treatment group and vehicle group (mean±SEM; P =0.09, t test). Note similarity to data observed in the sheep model (text, Figure 1 ). Right, Relationship between the extent of the ischemic risk zone (g) and the extent of necrosis (g) in combined Bendavia group and vehicle group. Regression lines: vehicle is indicated by dashed line; Bendavia, solid line.

Techniques Used: In Vivo

Relationship between risk zone (g) and the extent of no-reflow (g). Lines of regression for vehicle-treated rabbits (dashed line) and Bendavia-treated rabbits (solid line) are shown. * P =0.0085 by ANCOVA testing for a treatment effect (Bendavia vs vehicle) on the relationship.
Figure Legend Snippet: Relationship between risk zone (g) and the extent of no-reflow (g). Lines of regression for vehicle-treated rabbits (dashed line) and Bendavia-treated rabbits (solid line) are shown. * P =0.0085 by ANCOVA testing for a treatment effect (Bendavia vs vehicle) on the relationship.

Techniques Used:

Left, Ischemic risk zone (% LV) and infarct size (% risk zone) in sheep exposed to in vivo ischemia/reperfusion (60 min/3 h). Bendavia (0.05 mg/kg per hour; black bars) was administered intravenously beginning 30 minutes before reperfusion (mean±SEM; P =0.02, t test). Right, Relationship between the extent of the ischemic risk zone (% LV) and the extent of necrosis (% LV) ( P =0.03, ANCOVA).
Figure Legend Snippet: Left, Ischemic risk zone (% LV) and infarct size (% risk zone) in sheep exposed to in vivo ischemia/reperfusion (60 min/3 h). Bendavia (0.05 mg/kg per hour; black bars) was administered intravenously beginning 30 minutes before reperfusion (mean±SEM; P =0.02, t test). Right, Relationship between the extent of the ischemic risk zone (% LV) and the extent of necrosis (% LV) ( P =0.03, ANCOVA).

Techniques Used: In Vivo

Cellular ROS production during hypoxia/reoxygenation. A, Representative fluorescence images of cardiac ventricular myocytes loaded with the ROS sensor CM-DCF. ROS bursts during reoxygenation preceded cell death, and Bendavia treatment prevented ROS-induced cell death. B, Representative fluorescence intensity traces for cells in the study. CM-DCF fluorescence is normalized to the basal fluorescence (F o ) for each cell at the end of hypoxia. C, Contribution of ROS bursts to myocyte death during hypoxia/reoxygenation.
Figure Legend Snippet: Cellular ROS production during hypoxia/reoxygenation. A, Representative fluorescence images of cardiac ventricular myocytes loaded with the ROS sensor CM-DCF. ROS bursts during reoxygenation preceded cell death, and Bendavia treatment prevented ROS-induced cell death. B, Representative fluorescence intensity traces for cells in the study. CM-DCF fluorescence is normalized to the basal fluorescence (F o ) for each cell at the end of hypoxia. C, Contribution of ROS bursts to myocyte death during hypoxia/reoxygenation.

Techniques Used: Fluorescence

9) Product Images from "Impairment of glyoxalase-1, an advanced glycation end-product detoxifying enzyme, induced by inflammation in age-related osteoarthritis"

Article Title: Impairment of glyoxalase-1, an advanced glycation end-product detoxifying enzyme, induced by inflammation in age-related osteoarthritis

Journal: Arthritis Research & Therapy

doi: 10.1186/s13075-018-1801-y

Correlations between pentosidine, carboxymethyl-lysine (CML), and methylglyoxal-hydroimidazolone-1 (MG-H1) concentrations and patient age in human osteoarthritic cartilage. a MG-H1 quantified by liquid chromatography coupled to mass spectrometry. b Pentosidine quantified by liquid chromatography coupled to mass spectrometry. c CML quantified by liquid chromatography coupled to mass spectrometry. n = 11 in each experiment
Figure Legend Snippet: Correlations between pentosidine, carboxymethyl-lysine (CML), and methylglyoxal-hydroimidazolone-1 (MG-H1) concentrations and patient age in human osteoarthritic cartilage. a MG-H1 quantified by liquid chromatography coupled to mass spectrometry. b Pentosidine quantified by liquid chromatography coupled to mass spectrometry. c CML quantified by liquid chromatography coupled to mass spectrometry. n = 11 in each experiment

Techniques Used: Liquid Chromatography, Mass Spectrometry

10) Product Images from "Impairment of glyoxalase-1, an advanced glycation end-product detoxifying enzyme, induced by inflammation in age-related osteoarthritis"

Article Title: Impairment of glyoxalase-1, an advanced glycation end-product detoxifying enzyme, induced by inflammation in age-related osteoarthritis

Journal: Arthritis Research & Therapy

doi: 10.1186/s13075-018-1801-y

Correlations between pentosidine, carboxymethyl-lysine (CML), and methylglyoxal-hydroimidazolone-1 (MG-H1) concentrations and patient age in human osteoarthritic cartilage. a MG-H1 quantified by liquid chromatography coupled to mass spectrometry. b Pentosidine quantified by liquid chromatography coupled to mass spectrometry. c CML quantified by liquid chromatography coupled to mass spectrometry. n = 11 in each experiment
Figure Legend Snippet: Correlations between pentosidine, carboxymethyl-lysine (CML), and methylglyoxal-hydroimidazolone-1 (MG-H1) concentrations and patient age in human osteoarthritic cartilage. a MG-H1 quantified by liquid chromatography coupled to mass spectrometry. b Pentosidine quantified by liquid chromatography coupled to mass spectrometry. c CML quantified by liquid chromatography coupled to mass spectrometry. n = 11 in each experiment

Techniques Used: Liquid Chromatography, Mass Spectrometry

11) Product Images from "Evaluating the Suitability of Using Rat Models for Preclinical Efficacy and Side Effects with Inhaled Corticosteroids Nanosuspension Formulations"

Article Title: Evaluating the Suitability of Using Rat Models for Preclinical Efficacy and Side Effects with Inhaled Corticosteroids Nanosuspension Formulations

Journal: Nanoscale Research Letters

doi: 10.1007/s11671-010-9597-y

Ciclesonide nano particles
Figure Legend Snippet: Ciclesonide nano particles

Techniques Used:

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High Performance Liquid Chromatography:

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Article Snippet: .. LC/MS/MS Analysis All experiments were performed by using a API 5000 or API 4000 triple quadrupole mass spectrometer (AB/MDS Sciex, Concord, Canada) and an Agilent 1100 HPLC pump (Agilent, Andover, MA). .. Columns were Xbridge phenyl 2.1×100 mm (Waters, Milford, MA).

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Concentration Assay:

Article Title: CKD-506, a novel HDAC6-selective inhibitor, improves renal outcomes and survival in a mouse model of systemic lupus erythematosus
Article Snippet: .. The concentration of CKD-506 in samples were analyzed using API 4000 QTRAP (AB SCIEX) (Framingham, MA). .. Eighty female NZB/W F1 mice aged 4 weeks were purchased from Jackson Laboratory (Bar Harbor, ME, USA) and housed in SPF room of Samsung Biomedical Research Institute.

Mass Spectrometry:

Article Title: TPT sulfonate, a single, oral dose schistosomicidal prodrug: In vivo efficacy, disposition and metabolic profiling
Article Snippet: .. 2.7 Positive ion-mode electrospray tandem MS of TPT sulfonate and its metabolites in mouse plasma and mouse hepatocyte digests The principal fragments of m/z 290 TPT sulfonate observed using an API 4000 LC-MS/MS system (SCIEX, Framingham, MA) (see description under Results) are m/z 234, 135, 120 and 91. ..

Article Title: SCD1 Inhibition Causes Cancer Cell Death by Depleting Mono-Unsaturated Fatty Acids
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Article Title: Simultaneous analysis of herbicides pendimethalin, oxyfluorfen, imazethapyr and quizalofop-p-ethyl by LC–MS/MS and safety evaluation of their harvest time residues in peanut (Arachis hypogaea L.)
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Article Title: Modification of the IFCC reference measurement procedure for determination of HbA1c by HPLC-ESI-MS
Article Snippet: .. The mass spectrometer was an API 4000 equipped with a TurboV™ ESI source with Turboion Spray™ probe from Applied Biosystems (MDS-Sciex, Darmstadt, Germany). .. Sample preparation and calculation The samples were prepared according to the original IFCC reference measurement procedure [ ].

Liquid Chromatography with Mass Spectroscopy:

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Article Snippet: .. The organic layer was evaporated under a nitrogen stream, the dry residue was reconstituted in 100 μl of ACN/water (v/v, 1:4) and transferred to a LC/MS vial for analysis using an Applied Biosystems API 4000 MS/MS (AB Sciex) as described above. .. The mass spectrometer was operated in the positive ion electrospray mode, and multiple reaction monitoring transitions of m / z 256.0 > 158.8 for M2 and m / z 712.32 > 399.23 for ITZ- d 5 were monitored.

Tandem Mass Spectroscopy:

Article Title: Stereospecific Metabolism of Itraconazole by CYP3A4: Dioxolane Ring Scission of Azole Antifungals
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