Altered circadian rhythms and sleep in a new Angelman Syndrome mouse model

Normal neurodevelopment requires precise expression of the key ubiquitin ligase gene Ube3a. Comparing newly generated mouse models for Ube3a down-regulation (models of Angelman syndrome) vs. Ube3a up-regulation (models for autism), we find reciprocal effects of Ube3a gene dosage on phenotypes associated with circadian rhythmicity, including the amount of locomotor activity. In contrast to previous reports, we find that Ube3a is imprinted in neurons of the suprachiasmatic nuclei, the pacemaking circadian brain locus. In addition, Ube3a-deficient mice lack the typical drop in wake late in the dark period and have blunted responses to sleep deprivation. Suppression of physical activity by light in Ube3a-deficient mice is not due to anxiety as measured by behavioral tests and stress hormones; quantification of stress hormones may serve as an easily measurable biomarker for evaluating potential therapeutic treatments for Angelman syndrome. We conclude that reduced Ube3a gene dosage affects not only neurodevelopment but also sleep patterns and circadian rhythms.

Interplay between positive and negative regulation by B3-type transcription factors is critical for the accurate expression of the ABA INSENSITIVE 4 gene.

The ABA-INSENSITIVE 4 transcription factor is key for the regulation of diverse aspects of plant development and environmental responses, including proper perception of hormonal and nutritional signals. ABI4 activity is highly regulated at the transcriptional and post-transcriptional levels leading to precise expression mainly in the developing seed and early seedling development. Based on genetic and molecular approaches in the current study we provide new insights into the central mechanism underpinning the transcriptional regulation of ABI4 during both seed and vegetative development. We identified a complex interplay between the LEC2 and ABI3 transcriptional activators and the HSI/VAL repressors that is critical for proper ABI4 expression. Interestingly, the regulation by these proteins relies on the two RY cis-acting motifs present two kb upstream of the ABI4 gene. Our analysis also shows that the chromatin landscape of the ABI4 loci is highly dependent on the LEC2 and HSI2/VAL proteins. LEC2 regulation extends to the vegetative development and the absence of this factor results in ABA- and sugar-insensitive signaling in the developing plant. This regulatory circuit functions as a major control module for the correct spatial-temporal expression of ABI4 and prevents its ectopic accumulation that is harmful to the plant.

Dnmt3aa but Not Dnmt3ab Is Required for Maintenance of Gametogenesis in Nile Tilapia (Oreochromis niloticus)

Dnmt3a, a de novo methyltransferase, is essential for mammalian germ line DNA methylation. Only one Dnmt3a is identified in mammals, and homozygous mutants of Dnmt3a are lethal, while two Dnmt3a paralogs, dnmt3aa and dnmt3ab, are identified in teleosts due to the third round of genome duplication, and homozygous mutants of dnmt3aa and dnmt3ab are viable in zebrafish. The expression patterns and roles of dnmt3aa and dnmt3ab in gonadal development remain poorly understood in teleosts. In this study, we elucidated the precise expression patterns of dnmt3aa and dnmt3ab in tilapia gonads. Dnmt3aa was highly expressed in oogonia, phase I and II oocytes and granulosa cells in ovaries and spermatogonia and spermatocytes in testes, while dnmt3ab was mainly expressed in ovarian granulosa cells and testicular spermatocytes. The mutation of dnmt3aa and dnmt3ab was achieved by CRISPR/Cas9 in tilapia. Lower gonadosomatic index (GSI), increased apoptosis of oocytes and spermatocytes and significantly reduced sperm quality were observed in dnmt3aa−/− mutants, while normal gonadal development was observed in dnmt3ab−/− mutants. Consistently, the expression of apoptotic genes was significantly increased in dnmt3aa−/− mutants. In addition, the 5-methylcytosine (5-mC) level in dnmt3aa−/− gonads was decreased significantly, compared with that of dnmt3ab−/− and wild type (WT) gonads. Taken together, our results suggest that dnmt3aa, not dnmt3ab, plays important roles in maintaining gametogenesis in teleosts.

An open-source semi-automated robotics pipeline for embryo immunohistochemistry

A significant challenge for developmental systems biology is balancing throughput with controlled conditions that minimize experimental artifacts. Large-scale developmental screens such as unbiased mutagenesis surveys have been limited in their applicability to embryonic systems, as the technologies for quantifying precise expression patterns in whole animals has not kept pace with other sequencing-based technologies. Here, we outline an open-source semi-automated pipeline to chemically fixate, stain, and 3D-image Drosophila embryos. Central to this pipeline is a liquid handling robot, Flyspresso, which automates the steps of classical embryo fixation and staining. We provide the schematics and an overview of the technology for an engineer or someone equivalently trained to reproduce and further improve upon Flyspresso, and highlight the Drosophila embryo fixation and colorimetric or antibody staining protocols. Additionally, we provide a detailed overview and stepwise protocol for our adaptive-feedback pipeline for automated embryo imaging on confocal microscopes. We demonstrate the efficiency of this pipeline compared to classical techniques, and how it can be repurposed or scaled to other protocols and biological systems. We hope our pipeline will serve as a platform for future research, allowing a broader community of users to build, execute, and share similar experiments.

Study on Design of Non-Circular Gears for Speed Control of the Squid Belly Opening and Gutting Machine (SBOGM)

Non-circular gears can maintain rotational motions of general gears and implement all varying rotational motions of the cam. They adjust the angular velocity of driven gear according to operating conditions and make precise changes in angular motion. The design of non-circular gears has not been sufficiently studied because of their particularity and complex design methods unlike spur gears. In the gutting section of the Squid Belly Opening and Gutting Machine (SBOGM), spur gears generate rotational impact due to constant angular velocities, causing noise and equipment damage; so, efficiency should be improved by varying sectional angular velocity. Therefore, we derived pitch curves by selecting angular velocity ratio considering operating environments, and the tooth profile was designed by calculating module for each section according to radius through theorical analysis for precise expression of angular velocity ratio. To confirm reliability of design, angular velocity ratio and structural safety of designed non-circular gears were verified using, commercial software, ‘DAFUL 2020 R1′.

Exploring the Roles of HERC2 and the NEDD4L HECT E3 Ubiquitin Ligase Subfamily in p53 Signaling and the DNA Damage Response

Cellular homeostasis is governed by the precise expression of genes that control the translation, localization, and termination of proteins. Oftentimes, environmental and biological factors can introduce mutations into the genetic framework of cells during their growth and division, and these genetic abnormalities can result in malignant transformations caused by protein malfunction. For example, p53 is a prominent tumor suppressor protein that is capable of undergoing more than 300 posttranslational modifications (PTMs) and is involved with controlling apoptotic signaling, transcription, and the DNA damage response (DDR). In this review, we focus on the molecular mechanisms and interactions that occur between p53, the HECT E3 ubiquitin ligases WWP1, SMURF1, HECW1 and HERC2, and other oncogenic proteins in the cell to explore how irregular HECT-p53 interactions can induce tumorigenesis.

„W nieoczekiwanych chwilach empatii” – dydaktyka uważności wobec poezji Elizabeth Bishop

<p><span lang="EN-US">The article constitutes an examination of the didactic potential of the poetry and painting of Elizabeth Bishop. Unhurried exposure to her work teaches one a close look at reality, trust in everyday experience, attentiveness to precise expression and the communicability of feelings. Thanks to a modernist poetics indifferent to interpretation but focused on seizing instants of life, the poem and image reveal themselves and fulfill each other in expressing experience and emotion. The didactic of mindfulness permits us to wander the world of Bishop to teach us how to teach.</span></p>

On the Sharp Gårding Inequality for Operators with Polynomially Bounded and Gevrey Regular Symbols

In this paper, we analyze the Friedrichs part of an operator with polynomially bounded symbol. Namely, we derive a precise expression of its asymptotic expansion. In the case of symbols satisfying Gevrey estimates, we also estimate precisely the regularity of the terms in the asymptotic expansion. These results allow new and refined applications of the sharp Gårding inequality in the study of the Cauchy problem for p-evolution equations.

Metonymic Words and Expressions Characteristic of American English

When a particular metonymic word or expression is used, it is necessary to have certain cultural and historical knowledge in order to understand what is concealed within these metonymic expressions. Sometimes these metonymic expressions reveal the variety and characteristic features of a culture and give an idea of its being different from others. In the following paper the main intention is to analyze metonymic expressions in American English and to indicate variety that they bring to the speech of language carriers. Metonymy is also used in everyday language in order to give a more colorful and precise expression to ideas. Metonymies are approached as conceptual processes of extension, i.e. they are not so much relationships between words as relationships between concepts. It is interesting to note from a methodological point of view that while research on metaphor, cognitive or otherwise, has been able to focus on its object of matter without necessarily considering metonymy, things are quite different when metonymy comes under analysis. People with resembling cultural backgrounds and many similarities in common may share similar metonymic expressions, but for those who do not have the same cultural background it could sound like a challenge. Metonymy is accordingly a relevant linguistic device that plays a key role in the study of language and culture, and in understanding the speech of people from different cultures. The variety that these expressions cover may range from proper names to names of food and meals.

Precise Expression of Afmed15 Is Crucial for Asexual Development, Virulence, and Survival of Aspergillus fumigatus

ABSTRACT The rise of drug resistance in fungal pathogens is becoming a serious problem owing to the limited number of antifungal drugs available. Identifying and targeting factors essential for virulence or development unique to fungal pathogens is one approach to develop novel treatments for fungal infections. In this study, we present the identification and functional characterization of a novel developmental regulator in Aspergillus fumigatus, AfMed15, which contained a conserved Med15_fungal domain, as determined by screening of a mutant library that contained more than 2,000 hygromycin-resistant A. fumigatus transformants. Downregulating the expression of Afmed15 abolished the conidiation and decreased the fungal virulence in an insect model. Strikingly, the overexpression of Afmed15 caused fungal death accompanied by intensive autophagy. RNA sequencing of an Afmed15 overexpression strain revealed that altered gene expression patterns were associated with carbon metabolism, energy metabolism, and translation. Interestingly, the addition of metal ions could partially rescue fungal death caused by the overexpression of Afmed15, indicating that disordered ion homeostasis is a potential reason for the fungal death caused by the overexpression of Afmed15. Considering that the precise expression of Afmed15 is crucial for fungal development, virulence, and survival and that no ortholog was found in humans, Afmed15 is an ideal target for antifungal-drug development. IMPORTANCE The identification and characterization of regulators essential for virulence or development constitute one approach for antifungal drug development. In this study, we screened and functionally characterized Afmed15, a novel developmental regulator in A. fumigatus. We demonstrate that the precise transcriptional expression of Afmed15 is crucial for fungal asexual development, virulence, and survival. Downregulating the expression of Afmed15 abolished the conidiation and decreased the fungal virulence in an insect model. In contrast, the overexpression of Afmed15 caused fungal death accompanied by intensive autophagy. Our study provides a foundation for further studies to identify compounds perturbing the expression of Afmed15 that may be used for the prevention of invasive A. fumigatus infections.