Bisphenol F exposure affects mouse oocyte in vitro maturation through inducing oxidative stress and DNA damage.

Bisphenol F (BPF), a substitute for bisphenol A (BPA), is progressively used to manufacture various consumer products. Despite the established reproductive toxicity of BPF, the underlying mechanisms remain to elucidate. This in-vitro study deep in sighted the BPF toxicity on mouse oocyte meiotic maturation and quality. After treating oocytes with BPF (300 µM), the oocyte meiotic progression was blocked, accentuated by a reduced rate in the first polar body extrusion (PBE). Next, we illustrated that BPF induced α-tubulin hyper-acetylation disrupted the spindle assembly and chromosome alignment. Concurrently, BPF resulted in severe oxidative stress and DNA damage, which triggered the early apoptosis in mouse oocytes. Further, altered epigenetic modifications following BPF exposure were proved by increased H3K27me3 levels. Concerning the toxic effects on spindle structure, oxidative stress, and DNA damage in mouse oocytes, BPF toxicity was less severe to oocyte maturation and spindle structure than BPA and induced low oxidative stress. However, compared with BPA, oocytes treated with BPF were more prone to DNA damage, indicating not less intense or even more severe toxic effects of BPF than BPA on some aspects of oocytes maturation. In brief, the present study established that like wise to BPA, BPF could inhibit meiotic maturation and reduce oocyte quality, suggesting it is not a safe substitute for BPA.

New aspects of quantum electrodynamics on electronic structure and dynamics.

Application of Alpha-oscillator theory to quantum electrodynamics (QED) solves the mystery (Feynman) of the double-slit phenomenon involved in the foundation of quantum mechanics (QM). Even if with the same initial condition given, different spots on the screen can be predicted deterministically with no introduction of hidden variables. The interference pattern is similar to, but cannot be reproduced quantitatively by, that of the QM wave function, contrary to many-years-anticipation: a new prediction, awaiting experimental test over and above the Bohr-Einstein gedanken experiment. The general proof has already been published in Ref. [3a] and the concrete numerical algorithm of the extended normal mode technique for concrete trajectory of one electron in Ref. [3b]. In this article, (1) the new "interpretation" of the QED wave function is given in section "Interpretation of Wave Function in QED": the QED wave function used in the extended normal mode technique gives probability density distribution function of the initial values of trajectories. Moreover, (2) for the sake of demonstration of this new interpretation, the time-independent stationary state QM wave function is substituted to the QED wave function in section "Internal Self-Stress of Energetic Particles": the QED wave function is realized by internal self-stress revealed as energy density at the initial conditions. The renewed energy density is applied to study a unified scheme for generalized chemical reactivity. This is a new kind of chemical force acting in between electrons not in between nuclei. This paves a way for more advanced time-dependent simulation of electronic structure and dynamics in chemical reaction dynamics by tracing trajectories of many electrons. © 2018 Wiley Periodicals, Inc.

Fertility preservation for trans men: frozen-thawed in vitro matured oocytes collected at the time of ovarian tissue processing exhibit normal meiotic spindles.

PURPOSES: At the moment of sex reassignment surgery (SRS), the ovarian tissue is sometimes cryopreserved as fertility preservation option for female-to-male trans men, also called trans men. During this preparation, cumulus-oocyte-complexes (COCs) can be found and in vitro matured. It is not known if these oocytes are developmentally competent. In order to use these oocytes for fertility preservation and subsequent fertilization, a normal spindle structure before and after vitrification is necessary. METHODS: A total of 680 COCs were collected from trans men (n = 16) at the time of SRS and after testosterone treatment. The COCs were subjected to in vitro maturation and those that reached the metaphase II stage (MII) were collected and split into two groups; group 1 was immediately fixed for spindle staining and group 2 was first vitrified and warmed followed by spindle staining. Statistical analysis was performed by Fisher's exact test. RESULTS: After 48 h in vitro maturation, 38.1% of COCs were at MII stage. Those oocytes were split in two groups: (1) 126 MII oocytes in the noncryopreservation group and (2) 133 MII oocytes underwent cryopreservation through vitrification. The oocyte survival rate, after 2 h warming, was 67.7%. Both the noncryopreserved and the vitrified group showed comparable results concerning normal spindle structure and chromosomes alignment, 85.7% vs. 92.2% (P = 0.27). CONCLUSIONS: Spindle structure analysis and chromosomal alignment after vitrification seem normal in in vitro matured COCs collected during the tissue processing of ovaries in trans men at the time of SRS. The MII oocytes do not seem to be morphologically affected by prolonged testosterone treatment.

A Cell-Based Assay for Mitotic Spindle Orientation.

The regulation of mitotic spindle orientation is essential to ensure proper cell division and development (Kiyomitsua and Cheeseman Nat Cell Biol 14:311-317, 2012). For identification of potential spindle orientation regulators, determination of the mitotic spindle angle is a well-known but time-consuming procedure. Here we describe a simple and time-saving phenotypic screening assay for the identification of potential spindle orientation regulators. This screen is based on the analysis of monopolar mitotic spindle structures, which form upon inhibition of the mitotic kinesin Eg5/KSP by the small-molecule inhibitor dimethylenastron (DME) or similar compounds.

Quantification of three-dimensional spindle architecture.

Mitotic and meiotic spindles are microtubule-based structures to faithfully segregate chromosomes. Electron tomography is currently the method of choice to analyze the three-dimensional (3D) architecture of both types of spindles. Over the years, we have developed methods and software for automatic segmentation and stitching of microtubules in serial sections for large-scale reconstructions. 3D reconstruction of microtubules, however, is only the first step toward biological insight. The second step is the analysis of the structural data to derive measurable spindle properties. Here, we present a comprehensive set of techniques to quantify spindle parameters. These techniques provide quantitative analyses of specific microtubule classes and are applicable to a variety of tomographic reconstructions of spindles from different organisms.

Elastic 'tethers' connect separating anaphase chromosomes in a broad range of animal cells.

We describe the general occurrence in animal cells of elastic components ("tethers") that connect individual chromosomes moving to opposite poles during anaphase. Tethers, originally described in crane-fly spermatocytes, exert force on chromosome arms opposite to the direction the anaphase chromosomes move. We show that they exist in a broad range of animal cells. Thus tethers are previously unrecognised components of general mitotic mechanisms that exert force on chromosomes and they need to be accounted for in general models of mitosis in terms of forces on chromosomes and in terms of what their roles might be.

Xenopus laevis Kif18A is a highly processive kinesin required for meiotic spindle integrity.

The assembly and functionality of the mitotic spindle depends on the coordinated activities of microtubule-associated motor proteins of the dynein and kinesin superfamily. Our current understanding of the function of motor proteins is significantly shaped by studies using Xenopus laevis egg extract as its open structure allows complex experimental manipulations hardly feasible in other model systems. Yet, the Kinesin-8 orthologue of human Kif18A has not been described in Xenopus laevis so far. Here, we report the cloning and characterization of Xenopus laevis (Xl) Kif18A. Xenopus Kif18A is expressed during oocyte maturation and its depletion from meiotic egg extract results in severe spindle defects. These defects can be rescued by wild-type Kif18A, but not Kif18A lacking motor activity or the C-terminus. Single-molecule microscopy assays revealed that Xl_Kif18A possesses high processivity, which depends on an additional C-terminal microtubule-binding site. Human tissue culture cells depleted of endogenous Kif18A display mitotic defects, which can be rescued by wild-type, but not tail-less Xl_Kif18A. Thus, Xl_Kif18A is the functional orthologue of human Kif18A whose activity is essential for the correct function of meiotic spindles in Xenopus oocytes.

Using a quadratic parameter sinusoid model to characterize the structure of EEG sleep spindles.

Sleep spindles are essentially non-stationary signals that display time and frequency-varying characteristics within their envelope, which makes it difficult to accurately identify its instantaneous frequency and amplitude. To allow a better parameterization of the structure of spindle, we propose modeling spindles using a Quadratic Parameter Sinusoid (QPS). The QPS is well suited to model spindle activity as it utilizes a quadratic representation to capture the inherent duration and frequency variations within spindles. The effectiveness of our proposed model and estimation technique was quantitatively evaluated in parameter determination experiments using simulated spindle-like signals and real spindles in the presence of background EEG. We used the QPS parameters to predict the energy and frequency of spindles with a mean accuracy of 92.34 and 97.73% respectively. We also show that the QPS parameters provide a quantification of the amplitude and frequency variations occurring within sleep spindles that can be observed visually and related to their characteristic "waxing and waning" shape. We analyze the variations in the parameters values to present how they can be used to understand the inter- and intra-participant variations in spindle structure. Finally, we present a comparison of the QPS parameters of spindles and non-spindles, which shows a substantial difference in parameter values between the two classes.

ATM controls proper mitotic spindle structure.

The recessive ataxia-telangiectasia (A-T) syndrome is characterized by cerebellar degeneration, immunodeficiency, cancer susceptibility, premature aging, and insulin-resistant diabetes and is caused by loss of function of the ATM kinase, a member of the phosphoinositide 3-kinase-like protein kinases (PIKKs) family. ATM plays a crucial role in the DNA damage response (DDR); however, the complexity of A-T features suggests that ATM may regulate other cellular functions. Here we show that ATM affects proper bipolar mitotic spindle structure independently of DNA damage. In addition, we find that in mitosis ATM forms a complex with the poly(ADP)ribose (PAR) polymerase Tankyrase (TNKS) 1, the spindle pole protein NuMA1, and breast cancer susceptibility protein BRCA1, another crucial DDR player. Our evidence indicates that the complex is required for efficient poly(ADP)ribosylation of NuMA1. We find further that a mutant NuMA1 version, non-phosphorylatable at potential ATM-dependent phosphorylation sites, is poorly PARylated and induces loss of spindle bipolarity. Our findings may help to explain crucial A-T features and provide further mechanistic rationale for TNKS inhibition in cancer therapy.