Detection and Quantification of Nuclear Morphology Changes in Apoptotic Cells by Fluorescence Microscopy and Subsequent Analysis of Visualized Fluorescent Signals

Mandelkow, Robert; Gümbel, Denis; Ahrend, Hannes; Kaul, Anne; Zimmermann, Uwe; Burchardt, Martin; Stope, Matthias B.

doi:10.21873/anticanres.11560

Cited by 77 publications

(65 citation statements)

References 14 publications

(14 reference statements)

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“…Interestingly, the TUNEL assay confirmed and expanded the results of nuclear chromatin morphology analysis as statistical significances found on chromatin damage were reproduced on DNA fragmentation. These results allow to confirm the suitability of apoptosis analysis by nuclear chromatin morphology, as reported in other studies (Mandelkow et al, ), in the cumulus oophorus, a numerically limited cellular system consisting of cells aggregated together.…”

Section: Discussionsupporting

confidence: 88%

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The mycotoxin beauvericin induces oocyte mitochondrial dysfunction and affects embryo development in the juvenile sheep

Mastrorocco

Martino

Marzano

et al. 2019

Molecular Reproduction Devel

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Beauvericin (BEA) is a mycotoxin produced by Beauveria bassiana and Fusarium species recently reported as toxic on porcine oocyte maturation and embryo development. The aim of this study was to assess, in the juvenile sheep, whether its effects are due to alterations of oocyte and/or embryo bioenergetic/oxidative status.Cumulus-oocyte-complexes (COCs) were exposed to BEA during in vitro maturation (IVM), evaluated for cumulus cell (CC) apoptosis, oocyte maturation and bioenergetic/oxidative status or subjected to in vitro fertilization (IVF) and embryo culture (IVEC). Oocyte nuclear maturation and embryo development were assessed after Hoechst staining and CC apoptosis was analysed by terminal deoxynucleotidyl transferase-mediated dUTP nick-End labeling assay and chromatin morphology after Hoechst staining by epifluorescence microscopy. Oocyte and blastocyst bioenergetic/ oxidative status were assessed by confocal microscopy after mitochondria and reactive oxygen species labelling with specific probes. BEA showed various toxic effects, that is, short-term effects on somatic and germinal compartment of the COC (CCs and the oocyte) and long-term carry-over effects on developing embryos. In detail, at 5 µM, it significantly reduced oocyte maturation and immature oocytes showed increased late-stage (Type C) CC apoptosis and DNA fragmentation while matured oocytes showed unaffected CC viability but abnormal mitochondrial distribution patterns. At lower tested concentrations (3-0.5 µM), BEA did not affect oocyte maturation, but matured oocytes showed reduced mitochondrial activity. At low concentrations, BEA impaired embryo developmental capacity and blastocyst quality after IVF and IVEC. In conclusion, in the juvenile sheep, COC exposure to BEA induces CC apoptosis and oocyte mitochondrial dysfunction with negative impact on embryo development.

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Section: Discussionsupporting

confidence: 88%

“…To identify possible mechanisms underlying BEA‐induced reduction of MII rates, the incidence of apoptosis after COC exposure to BEA was investigated. Cell shrinking has been reported among morphological alterations of apoptosis (Mandelkow et al, ). It was found that BEA induces ooplasmic shrinking, even in MII oocytes.…”

Section: Discussionmentioning

confidence: 99%

The mycotoxin beauvericin induces oocyte mitochondrial dysfunction and affects embryo development in the juvenile sheep

Mastrorocco

Martino

Marzano

et al. 2019

Molecular Reproduction Devel

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“…The effect of AuNPs-PEG-RNase A on apoptosis induction was evaluated by DAPI staining assay. 38 Approximately 2×10 3 cells were seeded in 96-well cell culture plate. After incubation at 37°C for 24 hours, the cells were washed 3 times with PBS (NaCl, KCl, KH 2 PO 4 , and Na 2 HPO 4 , pH 7.4).…”

Section: Apoptosis Assay By Dapimentioning

confidence: 99%

PEGylated gold nanoparticles-ribonuclease induced oxidative stress and apoptosis in colorectal cancer cells

et al. 2019

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Introduction: Currently, drug-induced reactive oxygen species (ROS) mediating apoptosis pathway have extensively been investigated in designing effective strategies for colorectal cancer (CRC) chemotherapy. Bovine pancreatic ribonuclease A (RNase A) represents a new class of cytotoxic and non-mutagenic enzymes, and has gained more attention as a potential anticancer modality; however, the cytosolic ribonuclease inhibitors (RIs) restrict the clinical application of this enzyme. Nowadays, nanotechnology-based diagnostic and therapeutic systems have provided potential solutions for cancer treatments. Methods: In this study, the gold nanoparticles (AuNPs) were synthesized, stabilized by polyethylene glycol (PEG), functionalized, and covalently conjugated with RNase A. The physicochemical properties of engineered nanobiomedicine (AuNPs-PEG-RNase A) were characterized by scanning electron microscope (SEM), dynamic light scattering (DLS), and UV-vis spectrum. Then, its biological impacts including cell viability, apoptosis, and ROS production were evaluated in the SW-480 cells. Results: The engineered nanobiomedicine, AuNPs-PEG-RNase A, was found to effectively induce apoptosis in SW-480 cells and result in a significant reduction in cancer cell viability. Besides, the maximum production of ROS was obtained after the treatment of cells with an IC50 dose of AuNPs-PEG-RNase A. Conclusion: Based on the efficient ROS-responsiveness and the anticancer activity of RNase A of the engineered nanomedicine, this nanoscaled biologics may be considered as a potential candidate for the treatment of CRC.

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“…In U2-OS and MNNG/HOS OS cell lines, CAP treatment activated the apoptosis-specific proteases caspase-3 and caspase-7 [unpublished data]. Later in the apoptotic cascade, cell shrinking has been demonstrated by fluorescence dye stained OS cell nuclei [ 110 ]. Microscopic analysis revealed decreased nuclei measured by a reduced nuclear area and perimeter.…”

Section: Cap Effects On Os Cellsmentioning

confidence: 99%

Cold Atmospheric Plasma in the Treatment of Osteosarcoma

Gümbel

Bekeschus

Gelbrich

et al. 2017

IJMS

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Human osteosarcoma (OS) is the most common primary malignant bone tumor occurring most commonly in adolescents and young adults. Major improvements in disease-free survival have been achieved by implementing a combination therapy consisting of radical surgical resection of the tumor and systemic multi-agent chemotherapy. However, long-term survival remains poor, so novel targeted therapies to improve outcomes for patients with osteosarcoma remains an area of active research. This includes immunotherapy, photodynamic therapy, or treatment with nanoparticles. Cold atmospheric plasma (CAP), a highly reactive (partially) ionized physical state, has been shown to inherit a significant anticancer capacity, leading to a new field in medicine called “plasma oncology.” The current article summarizes the potential of CAP in the treatment of human OS and reviews the underlying molecular mode of action.

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Detection and Quantification of Nuclear Morphology Changes in Apoptotic Cells by Fluorescence Microscopy and Subsequent Analysis of Visualized Fluorescent Signals

Cited by 77 publications

References 14 publications

The mycotoxin beauvericin induces oocyte mitochondrial dysfunction and affects embryo development in the juvenile sheep

The mycotoxin beauvericin induces oocyte mitochondrial dysfunction and affects embryo development in the juvenile sheep

PEGylated gold nanoparticles-ribonuclease induced oxidative stress and apoptosis in colorectal cancer cells

Cold Atmospheric Plasma in the Treatment of Osteosarcoma

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