Could scoring tailed and dumbbell-shaped nuclei increase the sensitivity of micronucleus analysis as a biomarker of radiation exposure?

In the cytokinesis-block micronucleus (CBMN) assay, micronuclei (MNi), nucleoplasmic bridges (NPBs), and nuclear budding (NBUD) are the most commonly analysed morphological types of nuclear abnormalities. In contrast, tailed and dumbbell-shaped nucleus have historically received little attention in the CBMN assay. Interestingly, the incidence of tailed and dumbbell-shaped nuclei in lymphocytes is closely related with that of dicentric chromosomes or NPBs in the CBMN assay. To provide a better picture of the implications and significance of tailed and dumbbell-shaped nuclei as markers of radiation exposure, a literature review was performed in this study. Twenty articles were found in PubMed, PubMed Central, and manually searched. The articles were screened and those that met the inclusion criteria and did not meet the exclusion criteria were reviewed by all authors. At the end, nine articles were included. In conclusion, the assessment of in vivo tailed nuclei in blood smears and accounting for the occurrence of dumbbell-shaped nuclei in the CBMN assay can increase the sensitivity of the CBMN assay for biodosimetry involving a high dose exposure.

Low Dose-Rate Irradiation of Gamma-Rays-Induced Cytotoxic and Genotoxic Alterations in Peripheral Erythrocytes of p53-Deficient Medaka (Oryzias latipes)

Morphological alterations and nuclear abnormalities in fish erythrocytes have been used in many studies as bioindicators of environmental mutagens including ionizing radiation. In this study, adult Japanese medaka (Oryzias latipes) were irradiated with gamma rays at a low dose rate (9.92 μGy/min) for 7 days, giving a total dose of 100 mGy; and morphological alterations, nuclear abnormalities, and apoptotic cell death induced in peripheral erythrocytes were investigated 8 h and 7 days after the end of the irradiation. A variety of abnormalities, such as tear-drop cell, crenated cell, acanthocyte, sickled cell, micronucleated cell, eccentric nucleus, notched nucleus, and schistocyte, were induced in the peripheral erythrocytes of the wild-type fish, and a less number of abnormalities and apoptotic cell death were induced in the p53-deficient fish. These results indicate that low dose-rate chronic irradiation of gamma rays can induce cytotoxic and genotoxic effects in the peripheral erythrocytes of medaka, and p53-deficient medaka are tolerant to the gamma-ray irradiation than the wild type on the surface.

Impaired lamin localization to the nuclear envelope is responsible for nuclear damage in LMNA mutant iPSC-derived cardiomyocytes

The LMNA gene encodes the nuclear envelope proteins Lamins A and C, which comprise a major part of the nuclear lamina, provide mechanical support to the nucleus, and participate in diverse intracellular signaling. LMNA mutations give rise to a collection of diseases called laminopathies, including dilated cardiomyopathy (LMNA-DCM) and muscular dystrophies. Although nuclear deformities are a hallmark of LMNA-DCM, the role of nuclear abnormalities in the pathogenesis of LMNA-DCM remains incompletely understood. Using induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) from LMNA mutant patients and healthy controls, we show that LMNA mutant iPSC-CM nuclei have altered shape or increased size compared to healthy control iPSC-CM nuclei. The LMNA mutation exhibiting the most severe nuclear deformities, R249Q, additionally caused reduced nuclear stiffness and increased nuclear fragility. Importantly, for all cell lines, the degree of nuclear abnormalities corresponded to the degree of Lamin A/C and Lamin B1 mislocalization from the nuclear envelope. The mislocalization was likely due to altered assembly of Lamin A/C. Collectively, these results point to the importance of correct lamin assembly at the nuclear envelope in providing mechanical stability to the nucleus and illustrate that defects in nuclear lamina organization can contribute to the nuclear and cellular dysfunction in LMNA-DCM.

The Role of Emerin in Cancer Progression and Metastasis

It is commonly recognized in the field that cancer cells exhibit changes in the size and shape of their nuclei. These features often serve as important biomarkers in the diagnosis and prognosis of cancer patients. Nuclear size can significantly impact cell migration due to its incredibly large size. Nuclear structural changes are predicted to regulate cancer cell migration. Nuclear abnormalities are common across a vast spectrum of cancer types, regardless of tissue source, mutational spectrum, and signaling dependencies. The pervasiveness of nuclear alterations suggests that changes in nuclear structure may be crucially linked to the transformation process. The factors driving these nuclear abnormalities, and the functional consequences, are not completely understood. Nuclear envelope proteins play an important role in regulating nuclear size and structure in cancer. Altered expression of nuclear lamina proteins, including emerin, is found in many cancers and this expression is correlated with better clinical outcomes. A model is emerging whereby emerin, as well as other nuclear lamina proteins, binding to the nucleoskeleton regulates the nuclear structure to impact metastasis. In this model, emerin and lamins play a central role in metastatic transformation, since decreased emerin expression during transformation causes the nuclear structural defects required for increased cell migration, intravasation, and extravasation. Herein, we discuss the cellular functions of nuclear lamina proteins, with a particular focus on emerin, and how these functions impact cancer progression and metastasis.

Microplastics-Induced Eryptosis and Poikilocytosis in Early-Juvenile Nile Tilapia (Oreochromis niloticus)

This study aims to assess the impact of microplastics (MPs) on erythrocytes using eryptosis (apoptosis) and an erythron profile (poikilocytosis and nuclear abnormalities), considered to be novel biomarkers in Nile tilapia (Oreochromis niloticus). In this study, four groups of fish were used: The first was the control group. In the second group, 1 mg/L of MPs was introduced to the samples. The third group was exposed to 10 mg/L of MPs. Finally, the fourth group was exposed to 100 mg/L of MPs for 15 days, following 15 days of recovery. The fish treated with MPs experienced an immense rise in the eryptosis percentage, poikilocytosis, and nuclear abnormalities of red blood cells (RBCs) compared with the control group in a concentration-dependent manner. Poikilocytosis of MP-exposed groups included sickle cell shape, schistocyte, elliptocyte, acanthocyte, and other shapes. Nuclear abnormalities of the MPs-exposed groups included micronuclei, binucleated erythrocytes, notched, lobed, blebbed, and hemolyzed nuclei. After the recovery period, a greater percentage of eryptosis, poikilocytotic cells, and nuclear abnormalities in RBCs were still evident in the groups exposed to MPs when crosschecked with the control group. The results show concerning facts regarding the toxicity of MPs in tilapia.

Micronuclei and other nuclear abnormalities in phorate exposed fish, Channa punctatus

Aim: The present study aims to establish morphology-based nuclear abnormalities (NAs) including micronuclei (MN) as effective and sensitive genotoxic endpoint biomarkers in fish against the sub-lethal exposure of phorate. Methodology: Fish, Channa punctatus (35 ± 3.0 g; 14.5 ± 1.0 cm) were randomly exposed in two sets, treated group 1 and 2 (0.0375 mg l-1 and 0.075 mg l-1 of phorate, respectively) along with a simultaneous control (0 mg l-1). The blood was sampled after 30 days. Results: A significant (p < 0.05) induction in reactive oxygen species (ROS) coupled with elevated frequency of blood cells showing micronuclei along with the gross appearance of notched nuclei, curved nuclei, blebbed nuclei, kidney-shaped nuclei, V-shaped nuclei, nuclear buds, nucleoplasmic bridges, dumbbell nuclei, and condensed/rounded nuclei were recorded in a dose-dependent manner. Interpretation: A significant (p < 0.05) induction in reactive oxygen species (ROS) coupled with elevated frequency of blood cells showing micronuclei along with the gross appearance of notched nuclei, curved nuclei, blebbed nuclei, kidney-shaped nuclei, V-shaped nuclei, nuclear buds, nucleoplasmic bridges, dumbbell nuclei, and condensed/rounded nuclei were recorded in a dose-dependent manner.

Suitability of Nanoparticles to Face Benzo(a)pyrene-Induced Genetic and Chromosomal Damage in M. galloprovincialis. An In Vitro Approach

Benzo(a)pyrene (B(a)P) is a well-known genotoxic agent, the removal of which from environmental matrices is mandatory, necessitating the application of cleaning strategies that are harmless to human and environmental health. The potential application of nanoparticles (NPs) in the remediation of polluted environments is of increasing interest. Here, specifically designed NPs were selected as being non-genotoxic and able to interact with B(a)P, in order to address the genetic and chromosomal damage it produces. A newly formulated pure anatase nano-titanium (nano-TiO2), a commercial mixture of rutile and anatase, and carbon black-derived hydrophilic NPs (HNP) were applied. Once it had been ascertained that the NPs selected for the work did not induce genotoxicity, marine mussel gill biopsies were exposed in vitro to B(a)P (2 μg/mL), alone and in combination with the selected NPs (50 µg/mL nano-TiO2, 10 µg/mL HNP). DNA primary reversible damage was evaluated by means of the Comet assay. Chromosomal persistent damage was assessed on the basis of micronuclei frequency and nuclear abnormalities by means of the Micronucleus-Cytome assay. Transmission Electron Microscopy (TEM) was performed to investigate the mechanism of action exerted by NPs. Pure Anatase n-TiO2 was found to be the most suitable for our purpose, as it is cyto- and genotoxicity free and able to reduce the genetic and chromosomal damage associated with exposure to B(a)P.