Almost invisible : a review of inclusion of LGBTQI people with cancer in online patient information resources

Objective: This review assessed the inclusion of lesbian, gay, bisexual, trans, queer and/or intersex (LGBTQI) people in online cancer information. Methods: The websites of Australian cancer organizations were reviewed to identify if they included LGBTQI people and the extent and nature of this inclusion. Websites that did not include LGBTQI people were then reviewed to identify if information was implicitly LGBTQI inclusive. International LGBTQI cancer information resources were reviewed to identify key content. Results: Of sixty-one Australian cancer organization websites reviewed, eight (13%) mentioned LGBTQI people, including 13 information resources targeted to LGBTQI people and 19 general cancer information resources that mentioned LGBTQI people. For Australian cancer websites that did not mention LGBTQI people, 88% used gender neutral language to refer to partners, 69% included a range of sexual behaviours, 13% used gender neutral language when referring to hormones or reproductive anatomy but none acknowledged diverse relationship types. Internationally, 38 LGBTQI-specific cancer information resources were identified. Conclusions: Cancer patient information resources need to be LGBTQI inclusive. LGBTQI-targeted resources are required to address this population's unique needs and improve cultural safety and cancer outcomes. Practice implications: Recommendations are provided for LGBTQI inclusive cancer patient information resources

The Science Performance of JWST as Characterized in Commissioning

This paper characterizes the actual science performance of the James Webb Space Telescope (JWST), as determined from the six month commissioning period. We summarize the performance of the spacecraft, telescope, science instruments, and ground system, with an emphasis on differences from pre-launch expectations. Commissioning has made clear that JWST is fully capable of achieving the discoveries for which it was built. Moreover, almost across the board, the science performance of JWST is better than expected; in most cases, JWST will go deeper faster than expected. The telescope and instrument suite have demonstrated the sensitivity, stability, image quality, and spectral range that are necessary to transform our understanding of the cosmos through observations spanning from near-earth asteroids to the most distant galaxies.Comment: 5th version as accepted to PASP; 31 pages, 18 figures; https://iopscience.iop.org/article/10.1088/1538-3873/acb29

Electrical synapse structure requires distinct isoforms of a postsynaptic scaffold.

Electrical synapses are neuronal gap junction (GJ) channels associated with a macromolecular complex called the electrical synapse density (ESD), which regulates development and dynamically modifies electrical transmission. However, the proteomic makeup and molecular mechanisms utilized by the ESD that direct electrical synapse formation are not well understood. Using the Mauthner cell of zebrafish as a model, we previously found that the intracellular scaffolding protein ZO1b is a member of the ESD, localizing postsynaptically, where it is required for GJ channel localization, electrical communication, neural network function, and behavior. Here, we show that the complexity of the ESD is further diversified by the genomic structure of the ZO1b gene locus. The ZO1b gene is alternatively initiated at three transcriptional start sites resulting in isoforms with unique N-termini that we call ZO1b-Alpha, -Beta, and -Gamma. We demonstrate that ZO1b-Beta and ZO1b-Gamma are broadly expressed throughout the nervous system and localize to electrical synapses. By contrast, ZO1b-Alpha is expressed mainly non-neuronally and is not found at synapses. We generate mutants in all individual isoforms, as well as double mutant combinations in cis on individual chromosomes, and find that ZO1b-Beta is necessary and sufficient for robust GJ channel localization. ZO1b-Gamma, despite its localization to the synapse, plays an auxiliary role in channel localization. This study expands the notion of molecular complexity at the ESD, revealing that an individual genomic locus can contribute distinct isoforms to the macromolecular complex at electrical synapses. Further, independent scaffold isoforms have differential contributions to developmental assembly of the interneuronal GJ channels. We propose that ESD molecular complexity arises both from the diversity of unique genes and from distinct isoforms encoded by single genes. Overall, ESD proteomic diversity is expected to have critical impacts on the development, structure, function, and plasticity of electrical transmission

ZO1b-Beta is necessary for robust Connexin localization to electrical synapses.

A-J. Confocal images of Mauthner circuit neurons and stereotypical electrical synaptic contacts in 5 days post fertilization (dpf) zf206Et zebrafish larvae from wt (A, B), tjp1b/ZO1b-pan-/- (C, D), tjp1b/ZO1b-alpha-/- (E, F), tjp1b/ZO1b-beta-/- (G, H), and tjp1b/ZO1b-gamma-/- (I, J) animals. Animals are stained with anti-GFP (green), anti-Cx35.5 (cyan), anti-Cx34.1 (yellow), and anti-ZO1 (magenta). Scale bars = 2 μm. Boxed regions denote stereotyped location of electrical synapses and regions are enlarged in neighboring panels. Images of the Mauthner cell body and lateral dendrite in the hindbrain (A, C, E, G, I) are maximum intensity projections of ~10–20 μm. In A’, C’, E’, G’, and I’, images are maximum-intensity projections of ~3–6 μm and neighboring panels show the individual channels. Images of the sites of contact of M/CoLo processes in the spinal cord (B, D, F, H, J) are maximum-intensity projections of ~6–8 μm. In B’, D’, F’, H’ and J’, images are from a single 0.42 μm Z-plane and the white dashed circle denotes the location of the M/CoLo site of contact. Neighboring panels show individual channels. Anterior up.</p

Quantification of ZO1 isoform mutants.

A. Quantification of Cx35.5, Cx34.1, and ZO1 fluorescence intensities at CE synapses for the noted genotypes. The height of the bar represents the mean of the sampled data normalized to the wt average. Circles represent the normalized value of each individual animal. Mean is shown ± SEM. wt n = 10; tjp1b/ZO1b-pan-/- n = 5, tjp1b/ZO1b-alpha-/- n = 5, tjp1b/ZO1b-beta-/- n = 10, and tjp1b/ZO1b-gamma-/- n = 5. For Cx35.5 (cyan circles), **** indicates pB. Quantification of Cx35.5, Cx34.1, and ZO1 fluorescence intensities at M/CoLo synapses for the noted genotypes. The height of the bar represents the mean of the sampled data normalized to the wt average. Circles represent the normalized value of each individual animal. Mean is shown ± SEM. wt n = 11, tjp1b/ZO1b-pan-/- n = 3, tjp1b/ZO1b-alpha-/- n = 5, tjp1b/ZO1b-beta-/- n = 5, and tjp1b/ZO1b-gamma-/- n = 5. For Cx35.5 (cyan circles), **** indicates p<0.0001 by ANOVA with Dunnett’s test. For Cx34.1 (yellow circles), **** indicates p<0.0001 by ANOVA with Dunnett’s test. For ZO1 (magenta circles), **** indicates p<0.0001, ** indicates p = 0.008, and * indicates p = 0.0364 by ANOVA with Dunnett’s test.</p

Model of <i>tjp1b/ZO1b</i> isoform contribution to electrical synapse assembly.

Diagram summarizing the contribution of each ZO1b isoform to electrical synapse formation in the Mauthner circuit. Unique first exons of tjp1b/ZO1b-Alpha (black), -Beta (magenta), and -Gamma (blue) are denoted by triangles on the tjp1b-ZO1b locus (grey bar) with relative exon structure denoted (dark grey rectangles). Curved arrows indicate unique transcription initiation sites. ZO1b-Beta and -Gamma both localized to the electrical synapse, yet only the ZO1b-Beta isoform is required for robust Connexin (grey ovals) localization to synaptic contacts.</p

Summary of transgenic fish generated and used in this study.

A. A list of single mutant, double mutant, and V5-affinity tagged animals. Columns detail the tjp1b/ZO1b isoform(s) targeted for edit, the exon location and mutation recovered, the background in which the animal was generated, and the line designation assigned. (TIF)</p

V5-ZO1b-Beta and V5-ZO1b-Gamma isoform expression.

A. Detection of V5-N-terminal tag integration by CRISPR/Cas9 mediated HDR using a short, single-stranded nucleotide repair oligo. Genomic DNA prepared from individual injected 5 dpf zebrafish was analyzed for successful integration by PCR using a forward primer against the V5 tag and a reverse primer outside the modified region. Products for V5-Alpha (top), V5-Beta (middle) and V5-Gamma (bottom) were resolved by agarose gel electrophoresis (indicated by black arrows). Non-specific products are indicated with an asterisk (*). U = uninjected siblings, NTC = no template control. The table shows the percentage of siblings positive for integration and the percentage of siblings positive for V5 immunostain at any body location in the animal, indicating mosaic expression of V5-tagged ZO1b isoforms. B,C. Confocal images of the sites of contact of Mauthner/CoLo processes in the spinal cord of 5 dpf zebrafish larvae mosaically expressing V5-tjp1b/ZO1b-Beta (B) and V5-tjp1b/ZO1b-Gamma (C). V5-tjp1b/ZO1b-Beta animals are stained with anti-Cx35.5 (white), and anti-V5 (magenta). V5-tjp1b/ZO1b-Gamma animals are stained with anti-Cx36 (white), and anti-V5 (magenta). Anterior up. Scale bars = 2 μm. Images are maximum-intensity projections of ~3–4 μm and the dashed circle denotes the M/CoLo site of contact. Neighboring panels show individual channels. D,E. Confocal images of spinal cord floor plate collected from heterozygous V5-tjp1b/ZO1b-Beta (D) and heterozygous V5-tjp1b/ZO1b-Gamma (E) animals. Animals are stained with anti-V5 (magenta). Scale bars = 2 μm. Images are maximum-intensity projections of ~4 μm. Anterior left. F. Confocal tile scan of zebrafish brain from 5 dpf zf206Et zebrafish larvae from V5-tjp1b/ZO1b-Beta animals. Images are maximum intensity projections of ~42 μm. Animals are stained with anti-GFP (green), anti-V5 (magenta), and anti-Cx36 (white). Scale bars = 20 μm. Boxed region denotes stereotyped location of electrical synapses where V5-ZO1b-Beta and Cx36 overlap, and the region is enlarged in neighboring panels. White arrows denote regions where V5-ZO1b-Beta and Cx36 do not overlap. Anterior left. In F’, neighboring panels show individual channels. (TIF)</p