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Search for "flow cytometry" in Full Text gives 29 result(s) in Beilstein Journal of Organic Chemistry.
Non-peptide compounds from Kronopolites svenhedini (Verhoeff) and their antitumor and iNOS inhibitory activities
Beilstein J. Org. Chem. 2023, 19, 789–799, doi:10.3762/bjoc.19.59
- digestion solution, ground, and centrifuged to produce a cell suspension. Lymphocytes were isolated from the cell suspension using the Percoll method and were further enriched (via negative selection) to obtain naive CD8+ T cells. The enrichment effect and phenotype of the CD8+ cells were detected by flow
- cytometry (BD, USA). CD8+ T cells and Panc02-h7-GP-GFP cells were co-cultured with the corresponding concentrations of compounds or DMSO for 18 h. Fluorescence intensity was measured using a microplate reader (emission at 476 nm, excitation at 514 nm). Anti-inflammatory assay RAW264.7 (a mouse macrophage
Formal total synthesis of macarpine via a Au(I)-catalyzed 6-endo-dig cycloisomerization strategy
Beilstein J. Org. Chem. 2022, 18, 1589–1595, doi:10.3762/bjoc.18.169
- -inflammatory [4][5][6][7][8], insecticidal, fungicidal, etc [9]. In addition to the above-mentioned activities, macarpine was also used as a DNA probe for flow cytometry and fluorescence microscopy due to its fluorescent properties [10]. Despite some research on the activities of macarpine had been performed
The ethoxycarbonyl group as both activating and protective group in N-acyl-Pictet–Spengler reactions using methoxystyrenes. A short approach to racemic 1-benzyltetrahydroisoquinoline alkaloids
Beilstein J. Org. Chem. 2021, 17, 2716–2725, doi:10.3762/bjoc.17.183
- compounds 2a–f, 3a–c and 4 regarding their P-glycoprotein blocking potential. For that, we used the P-glycoprotein substrate calcein as reporter dye and analyzed intracellular calcein fluorescence of P-glycoprotein overexpressing VCR-R CEM cells by flow cytometry. An increase of the calcein-positive
- after substraction of the basal value. Flow cytometry P-glycoprotein activity was assessed using the reporter dye calcein acetoxymethyl ester (AM) as described previously [59]. To assess apoptosis, VCR-R CEM cells were seeded at 0.125 × 106 cells/well, incubated for 4 h and stimulated as indicated for
- 48 h. Apoptosis was analyzed by propidium iodide staining and flow cytometry as described by Nicoletti et al. [60]. All flow cytometry experiments were conducted on a BD FACS Canto II (BD Biosciences). Prominent synthetic approaches to 1-benzyltetrahydroisoquinolines: Bischler–Napieralski, Pictet
Enhanced target cell specificity and uptake of lipid nanoparticles using RNA aptamers and peptides
Beilstein J. Org. Chem. 2021, 17, 891–907, doi:10.3762/bjoc.17.75
- h later, the apical layer was removed, and the basolateral cells were washed, trypsinized, and resuspended in 1× PBS. Detection of Cy5 was measured by flow cytometry on a BD Accuri™ C6 device (Becton, Dickinson and Company, NJ), and the data was analyzed using FlowJo™ Version 10.7.1 (Becton
Automated high-content imaging for cellular uptake, from the Schmuck cation to the latest cyclic oligochalcogenides
Beilstein J. Org. Chem. 2020, 16, 2007–2016, doi:10.3762/bjoc.16.167
- employed to generate dose–response curves and to calculate the CP50 value of the transporters, described as the half-maximal cell penetration. In a CAPA, the final fluorescence response is usually recorded by flow cytometry [55][62]. In a HC CAPA, flow cytometry is replaced by HC automated microscopy
- , masking exclusively the mitochondria region, gave a small but significant increase to r2 = 0.983 (Figure 8b). Although optimized toward perfection with stable HGM cells, this increase demonstrated that the HC CAPA adds a precision that is overlooked with flow cytometry. The off-target staining of 26 in
- 0.983 quantified the increase in accuracy achieved moving from a cell-body mask to a mitochondrial mask. This difference revealed that even with fully optimized stable HGM cell lines, whole-cell analyses, such as flow cytometry, of the CAPA contain a small but nonnegligible error that can be removed
Potent hemithioindigo-based antimitotics photocontrol the microtubule cytoskeleton in cellulo
Beilstein J. Org. Chem. 2020, 16, 125–134, doi:10.3762/bjoc.16.14
- cycle arrest in the G2/M phase by preventing the completion of mitosis [1]. We examined cell cycle repartition by quantification of cellular DNA content via propidium iodide (PI) incorporation, which was analysed by flow cytometry (Supporting Information File 1, Figure S7). HeLa cells were treated for
Diversity-oriented synthesis of spirothiazolidinediones and their biological evaluation
Beilstein J. Org. Chem. 2019, 15, 2774–2781, doi:10.3762/bjoc.15.269
- spirothiazolidinediones via a [2 + 2 + 2] cyclotrimerization reaction and the derivatives were further functionalized through DA chemistry and click reaction. Using flow cytometry, it was shown for the first time that the new benzyl alcohol derivatives of thiazolidine-2,4-dione generated here are efficient apoptosis
- inducers in the HeLa, Hek293, U937, Jurkat, and K562 cell lines. Keywords: apoptosis; biologically active; [2 + 2 + 2] cycloaddition; flow cytometry; spiro thiazolidinedione; Introduction Heterocyclic compounds play a vital role in the metabolism of all living cells. Thus, most of the biologically active
- the culture of Jurkat cells at a concentration of 0.75 nM and was 68.65%. Under similar conditions, compound 20 showed about 95% in K562 cells of late apoptosis and compound 17 about 30.28% for cell line U937. The results of flow cytometry showed that in all three cell lines, Jurkat, K562 and U937, a
α,ß-Didehydrosuberoylanilide hydroxamic acid (DDSAHA) as precursor and possible analogue of the anticancer drug SAHA
Beilstein J. Org. Chem. 2019, 15, 2524–2533, doi:10.3762/bjoc.15.245
- 11b has been studied since it was the most cytotoxic among all. FITC (fluorescein-5-isothiocyanate)–annexin V/PI flow cytometry of HepG2 cells On HeLa cells, the efficacy of 11b was measured through the application of other apoptotic parameters like phosphatidylserine (PS) externalization, a known
- treated with 11b of 2.5 and 4.0 μg/mL concentrations at 37 °C for 24 h. Quantification of ROS by FACS Cells (2 × 105) were treated with SAHA, 11b, 11f, 11g at GI25, GI50 and GI75 concentrations for 24 h and the levels of intracellular ROS were assessed by flow cytometry after incubating with DCFH-DA (25
Cyclopropene derivatives of aminosugars for metabolic glycoengineering
Beilstein J. Org. Chem. 2019, 15, 584–601, doi:10.3762/bjoc.15.54
- although Ac4ManNCyoc reacts significantly faster in the DAinv reaction. To verify and to quantify these findings, we also analyzed the labeled cells by flow cytometry. We used the same conditions for MGE as described above (Scheme 1), but after incubation with strep-AF555, cells were released with trypsin
- , resuspended in buffer, and then subjected to flow cytometry analysis. The obtained results coincided with those of the fluorescence microscopy experiments (Figure 3B,C). Ac4ManNCyc gave a significantly higher fluorescence intensity than the negative control, which, however, is exceeded by far from that of
- conditions and microscopy settings resulted in a bright staining for Ac4GlcNCp and Ac4GalNCp well over that of the negative control (Figure 5). These results were confirmed by flow cytometry (Figure 4 and Figure 5). Interestingly, we did not observe cytotoxicity of Ac4GlcNCp up to a concentration of 100 μM
Impact of Pseudomonas aeruginosa quorum sensing signaling molecules on adhesion and inflammatory markers in endothelial cells
Beilstein J. Org. Chem. 2018, 14, 2580–2588, doi:10.3762/bjoc.14.235
- evaluated. The expression of adhesion molecules (VE-cadherin, PECAM-1, ICAM-1, and P-selectin) and inflammatory response molecules (IL-1β, IL-6, TNFα, TGFβ, and eNOS) was assessed by qRT-PCR and flow cytometry. Our results showed that bacterial adherence to inert substratum and biofilm were decreased in the
- the tested QSSMs is to disperse microbial aggregates attached on the host cells, since it is clearly seen that the untreated PAO1 control has a higher aggregative potential, as compared to all other samples (Figure 3). Flow cytometry analysis of endothelial cells adhesion molecules revealed that ICAM
- molecules, while independent, with similar or different effects, act together [22]. The expression of adhesion molecules VE-cadherin and PECAM-1 was evaluated by qRT-PCR assay, and the obtained results sustained the flow cytometry data, suggesting the ability of QSSMs to modulate adhesion of host cells. An
Comparative cell biological study of in vitro antitumor and antimetastatic activity on melanoma cells of GnRH-III-containing conjugates modified with short-chain fatty acids
Beilstein J. Org. Chem. 2018, 14, 2495–2509, doi:10.3762/bjoc.14.226
- conjugates and as the free drug was determined by flow cytometry (FACSCalibur; Becton Dickinson, San Jose, CA, USA). GeoMean (geometric mean channel) values normalized to the control are shown in Figure 2. The conjugates were internalized by A2058 cells in a time-dependent manner. In case of all conjugates
- conjugates could induce apoptosis and/or cell cycle arrest in melanoma cells. The pro-apoptotic effects of conjugates and Dau at 10−5 M concentration was studied after 24 h incubation by flow cytometry using FITC-annexin V (Sony Biotechnology, Weybridge, UK) and a novel image cytometer (NucleoCounter® NC
- cell cycle phases was analyzed by measurements of relative DNA contents of individual cells by flow cytometry after propidium iodide (Sigma-Aldrich, St. Louis, MO, USA) staining. The effects of conjugates and Dau on cell cycle phase distribution of A2058 cells are shown in Figure 4. The apoptotic
Natural and redesigned wasp venom peptides with selective antitumoral activity
Beilstein J. Org. Chem. 2018, 14, 1693–1703, doi:10.3762/bjoc.14.144
- low as 12.5 μmol L−1 for the selective targeting of MCF-7 breast cancer cells. Flow cytometry assays further revealed that treatment with wild-type (WT) peptide Dec-NH2 led to necrosis of MCF-7 cells. Additional atomic force microscopy (AFM) measurements indicated that the roughness of cancer cell
- L−1 after 2 h (Figure 2). Cell death assays Flow cytometry experiments were performed in an attempt to obtain insight into the mechanism of peptide-mediated death of cancer cells. For these proof-of-concept assays, we focused on WT peptide Dec-NH2. We utilized Annexin V labeling FITC (X axis) and
- analogs after 2 and 24 h of exposure to MCF-7 cancer cells. Experiments were done in triplicate. MTT assays evaluating the toxicity of Dec-NH2 and its derivatives towards MCF-10A normal cells after 2 and 24 h. Experiments were performed in triplicate. Cell death analysis using flow cytometry. Dot plot
Design and biological characterization of novel cell-penetrating peptides preferentially targeting cell nuclei and subnuclear regions
Beilstein J. Org. Chem. 2018, 14, 1378–1388, doi:10.3762/bjoc.14.116
- nucleolar region of the peptide NrTP alone could be confirmed also for NrTP-sC18* [3]. We then quantified the cellular uptake by using flow cytometry. As expected, the novel peptides N50-sC18* and NrTP-sC18* were characterized by an extremely high uptake compared to the CPP sC18*, as well as the nuclei
- solution (in acetate buffer) for 30 s. The stain was removed and the cells were washed twice with medium. After adding 300 μL of fresh medium, pictures were taken using a fluorescence confocal microscope. Images were edited in Image J 1.43m. Flow cytometry Cells were seeded in 24-well plates (HeLa 170,000
- min with 10 µM of CF-labeled peptide solutions. Green: CF-labeled peptide; blue: Hoechst 33342 nuclear stain; scale bar is 10 µm. Cellular uptake in MCF-7 and HeLa cells was quantified by flow cytometry. Cells were incubated with 10 µM peptide solutions for 30 min at 37 °C. Distribution pattern of the
Development of novel cyclic NGR peptide–daunomycin conjugates with dual targeting property
Beilstein J. Org. Chem. 2018, 14, 911–918, doi:10.3762/bjoc.14.78
- therefore the cellular uptake of Dau containing conjugates can be followed by flow cytometry (Figure 2). The new conjugates were taken up by HT-29 cells in a higher amount than by HT-1080 cells. Conjugates 2, 3 and 4 showed significantly higher accumulation in HT-29 than in HT-1080 cells compared to the
Synthesis and in vitro biochemical evaluation of oxime bond-linked daunorubicin–GnRH-III conjugates developed for targeted drug delivery
Beilstein J. Org. Chem. 2018, 14, 756–771, doi:10.3762/bjoc.14.64
- after 10 min). Additional flow cytometry studies demonstrated that the cellular uptake of the bioconjugate was inhibited in the presence of the competitive ligand triptorelin indicating a receptor-mediated pathway. For comparative purpose, six novel daunorubicin–GnRH-III bioconjugates have been
- understanding of the mechanism of action, the internalization and the intracellular localization, our lead compound K2 was studied and monitored by CLSM. Furthermore, the cellular uptake of K2 was evaluated in competition with the GnRH-I superagonist triptorelin by flow cytometry indicating a receptor mediated
- data of the conjugates cannot explain alone the results of their in vitro antitumor activity. Cellular uptake of the bioconjugates on MCF-7 human breast and HT-29 human colon cancer cells by flow cytometry The cellular uptake of the GnRH-III–drug conjugates was studied by flow cytometry on HT-29 and
Synthesis and biological evaluation of RGD and isoDGR peptidomimetic-α-amanitin conjugates for tumor-targeting
Beilstein J. Org. Chem. 2018, 14, 407–415, doi:10.3762/bjoc.14.29
- (breast adenocarcinoma) [32] were used as αVβ3 negative. The expression of αVβ3 integrin on the cell membrane was assessed by flow cytometry (see Supporting Information File 1, Figure S1), and the results were in good agreement with the literature for U87 and A549 [33][34][35]. In the case of MDA-MB-468
Halogen-containing thiazole orange analogues – new fluorogenic DNA stains
Beilstein J. Org. Chem. 2017, 13, 2902–2914, doi:10.3762/bjoc.13.283
- valuable properties as nucleic acid stains have made these dyes an irreplaceable tool in the active and developing area of bioanalytical chemical research [9][10]. Cyanine dyes have found various bioanalytical applications as soluble DNA intercalators (e.g., in living cell imaging [11] and flow cytometry
The chemistry and biology of mycolactones
Beilstein J. Org. Chem. 2017, 13, 1596–1660, doi:10.3762/bjoc.13.159
Glyco-gold nanoparticles: synthesis and applications
Beilstein J. Org. Chem. 2017, 13, 1008–1021, doi:10.3762/bjoc.13.100
- containing mannose-binding receptors) [46] and thyroglobulin in serum [47] and to study the internalization pathway in dendritic cells, employing confocal laser microscopy (CLSM) and flow cytometry [48]. Moreover, even techniques based on light scattering, like dark field microscopy (DFM), resulted
Expression, purification and structural analysis of functional GABA transporter 1 using the baculovirus expression system
Beilstein J. Org. Chem. 2017, 13, 874–882, doi:10.3762/bjoc.13.88
- system. GAT1/GFP recombinant proteins were expressed in Sf9 cells by viral infection for 72 h. The expression of the GAT1/GFP protein was further characterized. After 72 h post-infection, Sf9 cells with green fluorescence were controlled by flow cytometry (Figure 1A) and analyzed by fluorescence
- polyclonal antiserum. Subsequently, the blots were incubated with horseradish peroxidase-conjugated anti-rabbit antibody (IgG) (Dako Cytomation) and then visualized using amino ethylcarbazole (AEC) and substrate buffer (Calbiochem). Flow cytometry and fluorescence microscopy Sf9 cells were observed after 3
- -day infection by flow cytometry and fluorescence microscopy to determine the cell surface expression of the GAT1/GFP fusion proteins. MALDI mass fingerprinting Using MALDI–TOF MS, protein fragments with blocked N-termini or that are available in limited concentrations can be easily analyzed. The MALDI
Synthesis of spiro[isoindole-1,5’-isoxazolidin]-3(2H)-ones as potential inhibitors of the MDM2-p53 interaction
Beilstein J. Org. Chem. 2016, 12, 2793–2807, doi:10.3762/bjoc.12.278
- HepG2 cells). The LDH release was accompanied by a significant increase in cell death, as detected by flow cytometry through a propidium iodide assay (Figure 8b) [50][51]. Involvement of p53 in the pharmacological activity Tumor suppressor p53 plays an important role in conserving genome stability by
Smart molecules for imaging, sensing and health (SMITH)
Beilstein J. Org. Chem. 2015, 11, 2540–2548, doi:10.3762/bjoc.11.274
- microscopy and flow cytometry methods for preclinical research [13][14][15][16] (Figure 2). Many of these fluorescent probes are used as imaging reagents to quantify the level of cell death in a range of biomedical samples [17]. We also developed some nuclear isotopic labeled probes for in vivo imaging of
Cholesterol lowering effects of mono-lactose-appended β-cyclodextrin in Niemann–Pick type C disease-like HepG2 cells
Beilstein J. Org. Chem. 2015, 11, 2079–2086, doi:10.3762/bjoc.11.224
- reference wavelength of 630 nm was measured with a microplate reader (Bio-Rad Model 550, Tokyo, Japan). Cellular association of Lac-β-CyD Cellular association of Lac-β-CyD was determined by a flow cytometry. After incubation with TRITC-Lac-β-CyD for 1 h in U18666A-treated HepG2 cells, the cells were washed
Multivalent dendritic polyglycerolamine with arginine and histidine end groups for efficient siRNA transfection
Beilstein J. Org. Chem. 2015, 11, 763–772, doi:10.3762/bjoc.11.86
- maintaining its low cytotoxicity (Figure 4). Cellular uptake and confocal microscopy The cellular uptake and localization of fluorescently labeled siRNA/AAdPG complexes were quantified using flow cytometry and confocal microscopy (Figure 6). By comparing the cellular uptake of dPG-NH2 functionalized solely
Synthesis and biological evaluation of a novel MUC1 glycopeptide conjugate vaccine candidate comprising a 4’-deoxy-4’-fluoro-Thomsen–Friedenreich epitope
Beilstein J. Org. Chem. 2015, 11, 155–161, doi:10.3762/bjoc.11.15
- of tumor cells. Therefore, the binding of the induced antisera to MUC1-expressing MCF-7 human tumor cells was confirmed by flow cytometry using a fluorescently labelled goat anti-mouse-IgG antibody for visualization (see Supporting Information File 1). As shown in Figure 4, cells incubated with the
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