An accessible digital imaging workflow for multiplexed quantitative analysis of adult eye phenotypes in .

The compound eye of has long been a model for studying genetics, development, neurodegeneration, and heterochromatin. Imaging and morphometry of adult and other insects is hampered by the low throughput, narrow focal plane, and small image sensors typical of stereomicroscope cameras. When data collection is distributed among many individuals or extended time periods, these limitations are compounded by inter-operator variability in lighting, sample positioning, focus, and post-acquisition processing.

Sequence-Directed Covalent Protein-RNA Linkages in a Single Step Using Engineered HUH-Tags.

Replication-initiating HUH-endonucleases (Reps) are enzymes that form covalent bonds with single-stranded DNA (ssDNA) in a sequence specific manner to initiate rolling circle replication. These nucleases have been co-opted for use in biotechnology as sequence specific protein-ssDNA bioconjugation fusion partners dubbed 'HUH-tags'. Here, we describe the engineering and characterization of a series of laboratory evolved HUH-tags capable of forming robust sequence-directed covalent bonds with unmodified RNA substrates.

"Forcing" new interpretations of molecular tension sensor studies.

Molecular tension sensors are central tools for mechanobiology studies but have limitations in interpretation. Reporting in Cell Reports Methods, Shoyer et al. discover that fluorescent protein photoswitching in concert with sensor extension may expand the use and interpretation of common force-sensing tools.

HUHgle: An Interactive Substrate Design Tool for Covalent Protein-ssDNA Labeling Using HUH-Tags.

HUH-tags have emerged as versatile fusion partners that mediate sequence specific protein-ssDNA bioconjugation through a simple and efficient reaction. Here we present HUHgle, a python-based interactive tool for the visualization, design, and optimization of substrates for HUH-tag mediated covalent labeling of proteins of interest with ssDNA substrates of interest. HUHgle streamlines design processes by integrating an intuitive plotting interface with a search function capable of predicting and displaying protein-ssDNA bioconjugate formation efficiency and specificity in proposed HUH-tag/ssDNA sequence combinations.

Null regions: a unified conceptual framework for statistical inference.

Ruling out the possibility that there is absolutely no effect or association between variables may be a good first step, but it is rarely the ultimate goal of science. Yet that is the only inference provided by traditional null hypothesis significance testing (NHST), which has been a mainstay of many scientific fields. Reliance on NHST also makes it difficult to define what it means to replicate a finding, and leads to an uncomfortable quandary in which increasing precision in data reduces researchers' ability to perform theory falsification.

RAD-TGTs: high-throughput measurement of cellular mechanotype via rupture and delivery of DNA tension probes.

Mechanical forces drive critical cellular processes that are reflected in mechanical phenotypes, or mechanotypes, of cells and their microenvironment. We present here "Rupture And Deliver" Tension Gauge Tethers (RAD-TGTs) in which flow cytometry is used to record the mechanical history of thousands of cells exerting forces on their surroundings via their propensity to rupture immobilized DNA duplex tension probes. We demonstrate that RAD-TGTs recapitulate prior DNA tension probe studies while also yielding a gain of fluorescence in the force-generating cell that is detectable by flow cytometry.

Watson-Crick Base-Pairing Requirements for ssDNA Recognition and Processing in Replication-Initiating HUH Endonucleases.

Replication-initiating HUH endonucleases (Reps) are sequence-specific nucleases that cleave and rejoin single-stranded DNA (ssDNA) during rolling-circle replication. These functions are mediated by covalent linkage of the Rep to its substrate post cleavage. Here, we describe the structures of the endonuclease domain from the Muscovy duck circovirus Rep in complex with its cognate ssDNA 10-mer with and without manganese in the active site.

The Golden Age Is Behind Us: How the Status Quo Impacts the Evaluation of Technology.

New technology invariably provokes concerns over potential societal impacts. Even as risks often fail to materialize, the fear continues. The current research explored the psychological underpinnings of this pattern.

The Impact of Training in Transgender Care on Healthcare Providers Competence and Confidence: A Cross-Sectional Survey.

All studies to date demonstrate a lack of access to care for transgender people. A few educational efforts in providing care to transgender people have been successful. However, one challenge in administering training is that there is almost no research on the need of healthcare providers (HCP) to acquire knowledge, as well as on the effect of training on their level of competence and confidence in working with transgender people.

Information without knowledge: the effects of Internet search on learning.

The Internet has radically shifted how people access information. Instead of storing information internally, increasingly, people outsource to the Internet and retrieve it when needed. While this is an efficient strategy in many ways, its downstream consequences remain largely unexplored.

Health disparities between binary and non binary trans people: A community-driven survey.

Genderqueer and nonbinary () people have remained largely invisible in health research. Previous research shows worse outcomes on health indicators for trans people when compared with cisgender controls, but the differences between binary trans and GQNB individuals are inconclusive. To compare overall health and well-being of GQNB people with controls of trans men and trans women, taking into account the impact of the additive effect of their socio-economic position, as well as their current need for gender affirming medical interventions.

Depathologising gender diversity in childhood in the process of ICD revision and reform.

From 2007 on, the World Health Organisation (WHO) has been revising its diagnostic manual, the International Statistical Classification of Diseases and Related Health Problems (ICD), with approval of ICD-11 due in 2018. The ICD revision has prompted debates on diagnostic classifications related to gender diversity and gender development processes, and specifically on the 'Gender incongruence of childhood' (GIC) code. These debates have taken place at a time an emergent trans depathologisation movement is becoming increasingly international, and regional and international human rights bodies are recognising gender identity as a source of discrimination.