bluuebunny/crossref_metadata_2025 huggingface dataset:
AustralianCulex annulirostrismosquitoes are competent vectors for Japanese encephalitis virus genotype IV
Authors: Klein Melissa J. , Jackson Sarah A. , Suen Willy W. , Payne Jean , Beveridge Darcy , Hargreaves Mel , Gillies Donna , Wang Jianning , Blasdell Kim R. , Dunn Mike , López-Denman Adam J. , Williams David T. , Paradkar Prasad N.
Published: June 2024

Japanese encephalitis virus (JEV) is transmitted byCulexspecies of mosquitoes. In 2022, JEV belonging to a previously unrecognised lineage of genotype IV (GIV) caused a major outbreak of JE in South-eastern Australia, resulting in human cases and affecting piggeries.Cx. annulirostrishas previously been implicated as the major vector of JEV in northern Australia where the virus has circulated since its first detection in 1995. Here, we showed that experimental infection of a laboratory colony of AustralianCx. annulirostriswith the isolate JEV NSW/22 resulted in a 100% mosquito infection rate, with 87% of mosquito saliva samples testing positive by RT-qPCR at 14 days post-infection. Immunohistochemistry confirmed the presence of a replicating virus in the mosquito midgut and dissemination throughout the body, including the salivary glands. Our results also showed evidence of transovarial transmission of this virus; however, transstadial transmission from the eggs to the adult stage was not found. Comparison with an Indonesian isolate of GIV JEV and previous Australian isolates belonging to genotypes I and II showed that infection with JEV NSW/22 resulted in higher viral titres in the early stage of infection and higher proportions of mosquitoes with JEV-positive saliva, indicating a greater transmission potential compared to other isolates. This study provides compelling experimental evidence that AustralianCx. annulirostrisis a highly efficient vector for the 2022 Australian JEV GIV outbreak strain.
Parasitoid wasp venoms degradeDrosophilaimaginal discs for successful parasitism
Authors: Kamiyama Takumi , Shimada-Niwa Yuko , Mori Hitoha , Tani Naoki , Takemata-Kawabata Hitomi , Fujii Mitsuki , Takasu Akira , Katayama Minami , Kuwabara Takayoshi , Seike Kazuki , Matsuda-Imai Noriko , Senda Toshiya , Katsuma Susumu , Nakamura Akira , Niwa Ryusuke
Published: June 2024

Parasitoid wasps, one of the most diverse and species-rich animal taxa on Earth, produce venoms that manipulate host development and physiology to exploit host resources. However, mechanisms of venom action remain poorly understood. Here, we show that infection of hostDrosophilaby the endoparasitoid wasp,Asobara japonica, triggers imaginal disc degradation (IDD) by inducing apoptosis, autophagy, and mitotic arrest, leading to impaired host metamorphosis. A multi-omics approach identified two venom proteins ofA. japonicanecessary for IDD. Knockdown experiments targeting the venom genes revealed that in concert with host immune suppression, IDD is essential for successful parasitism. Our study highlights a venom-mediated hijacking strategy of the parasitoid wasp that allows host larvae to grow, but ultimately kills the hosts.
Poxvirus A52 is a host range factor for modified vaccinia virus Ankara (MVA) and promotes viral replication by disturbing the formation of autolysosomes
Authors: Niu Kang , Fang Yongxiang , Deng Yining , Wang Ziyue , Xie Shijie , Zhu Junda , Song Baifen , Wu Wenxue , Jing Zhizhong , Peng Chen
Published: June 2024

Many members of the poxvirus family are important zoonotic pathogens that pose a significant threat to human and animal health worldwide. Autophagy is a multi-step degradation pathway within cells, and one of its primary biological functions includes the clearance of invading viruses. Nevertheless, the interplay between poxviruses and host cell autophagy has not been fully elucidated. Here, we demonstrate that vaccinia virus (VACV) and lumpy skin disease virus (LSDV) induce incomplete autophagy and inhibit the fusion of autophagosomes and lysosomes, while modified vaccinia virus Ankara (MVA), an attenuated strain of VACV unable to replicate in almost all human cells, does not. Additionally, we screened and identified the VACV protein A52 as a key factor that obstruct the formation of autolysosomes. Mechanistically, A52 interacts with SNAP29 and inhibits its interaction with STX17 and VAMP8, both of which are binding partners of SNAP29 and are essential for complete autophagy. Moreover, A52 promotes the proteasomal degradation of SNAP29, which facilitates viral replication. We further revealed that SNAP29 functions as a restriction factor for MVA, as the suppression of SNAP29 allowed the replication of MVA in human cells. In summary, our data present a molecular mechanism by which poxviruses manipulate the cellular autophagic machinery and provide additional explanation for the restriction of MVA in human cells.
Is subjective perceptual similarity metacognitive?
Authors: Moharramipour Ali , Zhou William , Rahnev Dobromir , Lau Hakwan
Published: June 2024

Perceptual similarity is a cornerstone for human learning and generalization. However, in assessing the similarity between two stimuli differing in multiple dimensions, it is not well-defined which feature(s) one should focus on. The problem has accordingly been considered ill-posed. We hypothesize that similarity judgments may be, in a sense, metacognitive: The stimuli rated as subjectively similar are those that are in fact more challenging for oneself to discern in practice, in near-threshold settings (e.g., psychophysics experiments). This self-knowledge about one’s own perceptual capacities provides a quasi-objective ground truth as to whether two stimuli ‘should’ be judged as similar. To test this idea, we measure perceptual discrimination capacity between face pairs, and ask subjects to rank the similarity between them. Based on pilot data, we hypothesize a positive association between perceptual discrimination capacity and subjective dissimilarity, with this association being importantly specific to each individual.
Ultra-High Dose Rate Helium Ion Beams: Minimizing Brain Tissue Damage while Preserving Tumor Control
Authors: Dokic Ivana , Moustafa Mahmoud , Tessonnier Thomas , Meister Sarah , Ciamarone Federica , Akbarpour Mahdi , Krunic Damir , Haberer Thomas , Debus Jürgen , Mairani Andrea , Abdollahi Amir
Published: June 2024

Ultra-high dose rate radiotherapy with electrons and protons has shown potential for cancer treatment by effectively targeting tumors while sparing healthy tissues (FLASH effect). This study aimed to investigate the potential FLASH sparing effect of ultra-high-dose rate helium ion irradiation, focusing on acute brain injury and subcutaneous tumor response in a preclinical in vivo setting. Raster-scanned helium ion beams were used to compare the effects of standard dose rate (SDR at 0.2 Gy/s) and FLASH (at 141 Gy/s) radiotherapy on healthy brain tissue. Irradiation-induced brain injury was studied in C57BL/6 mice via DNA damage response, using nuclear γH2AX as a marker for double-strand breaks (DSB). The integrity of neurovascular and immune compartments was assessed through CD31+microvascular density and activation of microglia/macrophages. Iba1+ ramified and CD68+phagocytic microglia/macrophages were quantified, along with the expression of inducible nitric oxide synthetase (iNOS). Tumor response to SDR (0.2 Gy/s) and FLASH (250 Gy/s) radiotherapy was evaluated in A549 carcinoma model, using tumor volume and Kaplan-Meier survival as endpoints. The results showed that helium FLASH radiotherapy significantly reduced acute brain tissue injury compared to SDR, evidenced by lower levels of DSB and preserved neurovascular endothelium. Additionally, FLASH radiotherapy reduced neuroinflammatory signals compared to SDR, as indicated by fewer CD68+ iNOS+ microglia/macrophages. FLASH radiotherapy achieved tumor control comparable to that of SDR radiotherapy. This study is the first to report the FLASH sparing effect of raster scanning helium ion radiotherapy in vivo, highlighting its potential for neuroprotection and effective tumor control.
Kynurenic acid inflammatory signaling expands in primates and impairs prefrontal cortical cognition
Authors: Yang Shengtao , Datta Dibyadeep , Krienen Fenna M. , Ling Emi , Woo Elizabeth , May Athena , Anderson George M. , Galvin Veronica C. , Gonzalez-Burgos Guillermo , Lewis David A. , McCarroll Steven A. , Arnsten Amy FT , Wang Min
Published: June 2024

Cognitive deficits from dorsolateral prefrontal cortex (dlPFC) dysfunction are common in neuroinflammatory disorders, including long-COVID, schizophrenia and Alzheimer’s disease, and have been correlated with kynurenine inflammatory signaling. Kynurenine is further metabolized to kynurenic acid (KYNA) in brain, where it blocks NMDA and α7-nicotinic receptors (nic-α7Rs). These receptors are essential for neurotransmission in dlPFC, suggesting that KYNA may cause higher cognitive deficits in these disorders. The current study found that KYNA and its synthetic enzyme, KAT II, have greatly expanded expression in primate dlPFC in both glia and neurons. Local application of KYNA onto dlPFC neurons markedly reduced the delay-related firing needed for working memory via actions at NMDA and nic-α7Rs, while inhibition of KAT II enhanced neuronal firing in aged macaques. Systemic administration of agents that reduce KYNA production similarly improved cognitive performance in aged monkeys, suggesting a therapeutic avenue for the treatment of cognitive deficits in neuroinflammatory disorders.
A common neural signature between genetic and environmental risk
Authors: Vedechkina Maria , Holmes Joni , Warrier Varun , Astle Duncan
Published: June 2024

Not everyone is equally likely to experience mental illness. What is the contribution of an individual’s genetic background, or experiences of childhood adversity, to that likelihood? And how do these dimensions of risk interact at the level of the brain? We investigated the relationship between genetic liability for mental illness, childhood adversity, and cortico-limbic connectivity in a large developmental sample drawn from the ABCD cohort. First, we used Canonical Correlation Analysis to uncover two genetic dimensions of mental health using polygenic risk scores for ADHD, Anxiety, Depression, and Psychosis. The first dimension represented liability for broad psychopathology which positively correlated with adversity, while the second represented neurodevelopmental-specific risk which negatively interacted with adversity. Next, we investigated the cortico-limbic signature of adversity and genetic liability using Partial Least Squares. We found that the neural correlates of adversity broadly mirrored those of genetic liability, with adversity capturing most of the shared variance. These novel findings suggest that genetic and environmental riskoverlapin the neural connections that underlie behavioural symptomatology.
A novel monomeric amyloid β-activated signaling pathway regulates brain development via inhibition of microglia
Authors: Kwon Hyo Jun , Santhosh Devi , Huang Zhen
Published: June 2024

Amyloid β (Aβ) forms aggregates in the Alzheimer’s disease brain and is well known for its pathological roles. Recent studies show that it also regulates neuronal physiology in the healthy brain. Whether Aβ also regulates glial physiology in the normal brain, however, has remained unclear. In this article, we describe the discovery of a novel signaling pathway activated by the monomeric form of Aβ in vitro that plays essential roles in the regulation of microglial activity and the assembly of neocortex during development in vivo. We find that activation of this pathway depends on the function of amyloid precursor (APP) and the heterotrimeric G protein regulator Ric8a in microglia and inhibits microglial immune activation at transcriptional and post-transcriptional levels. Genetic disruption of this pathway during neocortical development results in microglial dysregulation and excessive matrix proteinase activation, leading to basement membrane degradation, neuronal ectopia, and laminar disruption. These results uncover a previously unknown function of Aβ as a negative regulator of brain microglia and substantially elucidate the underlying molecular mechanisms. Considering the prominence of Aβ and neuroinflammation in the pathology of Alzheimer’s disease, they also highlight a potentially overlooked role of Aβ monomer depletion in the development of the disease.
Multi-omics analyses and machine learning prediction of oviductal responses in the presence of gametes and embryos
Authors: Finnerty Ryan M. , Carulli Daniel J. , Hegde Akshata , Wang Yanli , Baodu Frimpong , Winuthayanon Sarayut , Cheng Jianlin , Winuthayanon Wipawee
Published: June 2024

The oviduct is the site of fertilization and preimplantation embryo development in mammals. Evidence suggests that gametes alter oviductal gene expression. To delineate the adaptive interactions between the oviduct and gamete/embryo, we performed a multi-omics characterization of oviductal tissues utilizing bulk RNA-sequencing (RNA-seq), single-cell RNA-sequencing (scRNA-seq), and proteomics collected from distal and proximal at various stages after mating in mice. We observed robust region-specific transcriptional signatures. Specifically, the presence of sperm induces genes involved in pro-inflammatory responses in the proximal region at 0.5 days post-coitus (dpc). Genes involved in inflammatory responses were produced specifically by secretory epithelial cells in the oviduct. At 1.5 and 2.5 dpc, genes involved in pyruvate and glycolysis were enriched in the proximal region, potentially providing metabolic support for developing embryos. Abundant proteins in the oviductal fluid were differentially observed between naturally fertilized and superovulated samples. RNA-seq data were used to identify transcription factors predicted to influence protein abundance in the proteomic data via a novel machine learning model based on transformers of integrating transcriptomics and proteomics data. The transformers identified influential transcription factors and correlated predictive protein expressions in alignment with thein vivo-derived data. Lastly, we found some differences between inflammatory responses in sperm-exposed mouse oviducts compared to hydrosalpinx fallopian tubes from patients. In conclusion, our multi-omics characterization and subsequentin vivoconfirmation of proteins/RNAs indicate that the oviduct is adaptive and responsive to the presence of sperm and embryos in a spatiotemporal manner.

Significance Statement

We conducted a detailed molecular study of how the oviduct changes its gene expression and protein production in response to sperm and embryos after mating in mice. We found that the oviduct has distinct molecular signatures in different regions – upper versus lower regions. Shortly after mating, inflammatory responses are turned on in the lower regions due to the presence of sperm. A day later, metabolic genes ramp up in the lower regions, likely to provide nutrients for the developing embryos. Overall, this multi-omics study revealed that the oviduct dynamically adapts its molecular makeup over time and space to accommodate and support sperm, eggs and embryos.
Children Who Develop Celiac Disease Exhibit Distinct Metabolic Pathways Among Their Gut Microbiota Years Before Diagnosis
Authors: Kelley Kristina , Dogru Dogus , Huang Qian , Yang Yi , Palm Noah W. , Ludvigsson Johnny , Altindis Emrah
Published: June 2024

Celiac disease (CD) is an autoimmune condition caused by a loss of tolerance to gluten in genetically predisposed individuals. While 30-40% of people possess the predisposing alleles, only 1-2% of the population is diagnosed with CD. This indicates environmental factors play a role in the pathogenesis of the disease, however the trigger of gluten tolerance loss is unknown. The gut microbiome composition of CD patients differs in comparison to their healthy counterparts; however, a causal link has not been established. In this study, we examined the alterations in the composition of the gut microbiota in a retrospective, longitudinal cohort of 10 children at age 1, matched for sex, human leukocyte antigen (HLA) genotype and breastfeeding duration. All samples were obtained from the pediatric donors prior to diagnosis (CD progressors). We used Ig-A sequencing combined with 16S sequencing for samples obtained at age 1. We also identified the functional metabolic pathways enriched in CD progressors compared to the healthy controls at ages 1, 2.5 (n=15-16) and 5 (n=9-13) using data from a similar study that we previously completed. Our findings demonstrate that CD progressors have ASV-level alterations in their gut microbiome as early as the first year of life, including the increased presence of some taxa that have been previously been reported to be enriched in CD. Using PICRUSt analysis, we also showed that inflammatory- and pathogenicity-related functions are enriched in CD progressors’ gut microbiome years before diagnosis. These pathways include glycine, serine and threonine metabolism, N-glycan biosynthesis and fatty acid biosynthesis and beta-lactam resistance, which are potentially contributing to chronic inflammation in CD. Overall, our results indicate distinct metabolic pathways enriched in the gut microbiome of CD progressors years before diagnosis. Understanding these pathways could advance our understanding of CD pathogenesis and its link to the gut microbiome.
Viral Insulin/IGF-like Peptides Selectively Activate Host Insulin/IGF Signaling pathways during Grouper Iridovirus Infection
Authors: Chuard Aurelien , Nesarajah Kalaimagal , Danazumi Khadija , Reiners Kaitlin , Zhang Fa , Levintov Lev , Vashisth Harish , Dimarchi Richard , Altindis Emrah
Published: June 2024

Until our recent discovery of viral insulin/IGF-1-like peptides (VILPs), examples of viral mimicry were mostly limited to immunomodulatory proteins and growth factors. We previously showed that six viruses in the Iridoviridae family encode genes mimicking insulin and IGF-1. While VILP-carrying viruses are isolated from fish, the role of VILPs in host-pathogen interactions remain elusive. In this study, we used the Grouper Iridovirus (GIV), one of the VILP-carrying viruses, to examine the impact of the GIV-VILPs on the host IGF-1R/IR system during infection. Our structural analysis revealed that GIV-VILP is 35% identical to both zebrafish insulin and 35 IGF-1. We also showed that GIV-VILPs are early viral transcripts and resulting peptides are secreted during viral infection. Both single-chain (sc, resembling IGF-1) and double chain (dc, resembling insulin) forms of chemically synthesized GIV-VILPs can stimulate insulin receptor (IR) and IGF-1 receptor (IGF1R) phosphorylation. They could also stimulatepost-receptor signaling in both grouper kidney (GK) and zebrafish AB.9 cells as potently as insulin. Notably, supernatants obtained from GIV-infected cells could stimulate insulin/IGF system in a dose dependent manner. Interestingly, GIV-VILP was able to selectively activate the Akt/PI3K pathway while had no or minimal effect on Erk/MAPK pathway. Using either IR, IGF1R or dual inhibitors, we illustrated that inhibition of IR signaling suppressed GIV viral replication while inhibition of IGF1R enhanced viral replication. Consistent with selective signaling action of PI3K/Akt pathway, inhibition of the Akt reduced GIV viral replication however Erk pathway inhibition did not affect it. In summary, GIV-VILP is produced during GIV infection and manipulates host insulin/IGF signaling in a selective manner. Our findings reveal a previously unknown viral strategy in which viruses mimic a host hormone to exploit the host’s endocrine system. This discovery unveils a novel viral pathogenesis mechanism, broadens our understanding of viral mimicry, and opens a new avenue to better understand viral strategies to manipulate the host.
Deriving connectivity from spiking activity in biophysical cortical microcircuits
Authors: Moghbel Faraz , Hassan Muhammad Taaha , Guet-McCreight Alexandre , Hay Etay
Published: June 2024

Inferring detailed cortical microcircuit connectivity is essential for uncovering how information is processed in the brain. A common methodin vivouses short-lag spike cross-correlations to derive putative monosynaptic connections between pairs of neurons, but previous studies did not address confounds of physiological large-scale networks such as correlated firing and inactive neurons. We tested connectivity derivation methods on ground-truth spiking data from detailed models of human cortical microcircuits in different layers and between key neuron types. We showed that physiological oscillations in the large-scale microcircuits hindered derivation accuracy, which was improved using a shorter cross-correlogram analysis window. We then showed that connection derivation was poor in cortical layer 2/3 microcircuits compared to layer 5, due to low firing rates and inactive neurons. General stimulation strategies for layer 2/3 microcircuits led to only a moderate improvement in derivation performance, due to a trade-off between the proportions of inactive neurons and overactive neurons, indicating the need for more refined strategies. Lastly, we showed that derivation of inhibitory connections from somatostatin interneurons targeting distal dendrites required a longer timescale of cross-correlation lags. Our results identify key physiological challenges and methods to improve accuracy in deriving connections from spiking activity in large-scale neuronal microcircuits.
Closely matched comparisons suggest that separable processes mediate contextual size illusions
Authors: Yu Xinran A. , Fischer Livia F. , Schwarzkopf Dietrich S.
Published: June 2024

Previous research suggests the magnitudes of the Ebbinghaus, Delboeuf, Ponzo, and tilt illusions all depend on the cortical distance between the neural representations of target stimuli and the surrounding context. However, several psychophysical studies found no compelling association between these illusions, calling this hypothesis into question. Here we ask if these discrepant reports could arise from methodological differences between these studies. We ran a battery of visual size illusion and basic discrimination tasks with carefully matched geometric properties, using a classic forced-choice design. In our small, homogenous sample, the Ebbinghaus and Delboeuf illusion magnitudes were strongly correlated, consistent with the idea that they reflect the same underlying mechanism when other sources of individual differences are minimised. Ponzo illusion magnitude also correlated with these two illusions, although less strongly in the case of the Ebbinghaus. Interestingly, the classic arrowhead version of the Mueller-Lyer illusion did not correlate with any of the other illusions or even with the objective ability to discriminate line length. This suggests that an altogether separate process underlies this perceptual effect. We further demonstrate that presenting stimuli briefly with central fixation critically affects measurements of the Ebbinghaus illusion. Additionally, we found that measuring illusion magnitude via adjustment is less reliable compared to two-alternative forced-choice procedures. Taken together, our findings suggest that different tasks probe separable processes determining illusion measurements. They further highlight the importance of the experimental design when testing relationships between perceptual effects and their links to neural processing.
Optimization of Soluble Expression of CTA1-(S14P5)4-DD and CTA1-(S21P2)4-DD Fusion Proteins as Candidates for COVID-19 Intranasal Vaccines
Authors: Tarigan Simson , Sumarningsih  , Sekarmila Gita , Apas  , Tarigan Ronald , Putri Ryandini , Setyawati Damai Ria
Published: June 2024

Developing intranasal vaccines against pandemics and devastating airborne infectious diseases is imperative. The superiority of intranasal vaccines over injectable systemic vaccines is evident, but the challenge in developing effective intranasal vaccines is more substantial. Fusing a protein antigen with the catalytic domain of cholera toxin (CTA1) and the two-domain D of staphylococcal protein A (DD) has significant potential for intranasal vaccines. In the present study, we constructed two fusion proteins containing CTA1, tandem repeat linear epitopes of the SARS-CoV-2 spike protein (S14P5 or S21P2), and DD. Thein silicocharacteristics and solubility of the fusion proteins CTA1-(S14P5)4-DD and CTA1-(S21P2)4-DD were analyzed when overexpressed inEscherichia coli. Structural predictions indicated that each component of the fusion proteins was compatible with its origin. Both fusion proteins were predicted by computational tools to be soluble when overexpressed inE. coli. Contrary to these predictions, the constructs exhibited limited solubility. The solubility did not improve even after lowering the cultivation temperature from 37°C to 18°C. Induction with IPTG at the early log phase, instead of the usual mid-log phase growth, significantly increased soluble CTA1-(S21P2)4-DD but not CTA1-(S14P5)4-DD. The solubility of overexpressed fusion proteins significantly increased when a non-denaturing detergent (Nonidet P40, Triton X100, or Tween 20) was added to the extraction buffer. In a scale-up purification experiment, the yields were low, only 1-2 mg/L of culture, due to substantial losses during the purification stages, indicating the need for further optimization of the purification process.
Pseudomonastaxonomic and functional microdiversity in the wheat rhizosphere is cultivar- dependent and links to disease resistance profile and root diameter
Authors: Herms Courtney Horn , Hennessy Rosanna Catherine , Bak Frederik , Guan Ying , Browne Patrick Denis , Nielsen Tue Kjærgaard , Hansen Lars Hestbjerg , Dresbøll Dorte Bodin , Nicolaisen Mette Haubjerg
Published: June 2024

Diversity within lower taxonomic units in microbial communities is a key trait, giving rise to important ecological functions. In the rhizosphere, these functions include disease suppression and pathogen inhibition. However, limited effort has been given to exploring intragenus microdiversity in an increasingly homogenous agricultural system. Through an integrative approach combining culture-dependent and -independent methods, we explore the rhizospherePseudomonaspangenome and demonstrate cultivar-dependent taxonomic and functional microdiversity between two closely related modern winter wheat cultivars. AFusarium-resistant cultivar demonstrated increasedPseudomonastaxonomic diversity but not biosynthetic diversity when compared to the susceptible cultivar, coinciding with a thinner root diameter of the resistant cultivar. We found enrichment ofPseudomonasisolates capable of antagonizingFusariumas well as chitinase-encoding genes and pyoverdine gene clusters in the resistant cultivar. Across closely relatedPseudomonasisolates from the two cultures, there were differences in genomic content and biosynthetic gene clusters. Ultimately, we highlight the need for fine-scale analysis to uncover the hidden microdiversity within rhizospherePseudomonas.
Heterogeneity in establishment of polyethylene glycol-mediated plasmid transformations for five forest pathogenicPhytophthoraspecies
Authors: Dort Erika N. , Hamelin Richard C.
Published: June 2024

Plasmid-mediated DNA transformation is a foundational molecular technique and the basis for most CRISPR-Cas9 gene editing systems. While plasmid transformations are well established for many agriculturalPhytophthorapathogens, development of this technique in forest Phytophthoras is lacking. Given our long-term research objective to develop CRISPR-Cas9 gene editing in a forest pathogenicPhytophthoraspecies, we sought to establish the functionality of polyethylene glycol (PEG)-mediated plasmid transformation in five species:P. cactorum,P. cinnamomi,P. cryptogea,P. ramorum, andP. syringae. We used the agricultural pathogenP. sojae, a species for which PEG-mediated transformations are well-established, as a transformation control. Using a protocol previously optimized forP. sojae, we tested transformations in the five forest Phytophthoras with three different plasmids: two developed for CRISPR-Cas9 gene editing and one developed for fluorescent protein tagging. Out of the five species tested, successful transformation, as indicated by stable growth of transformants on a high concentration of antibiotic selective growth medium and diagnostic PCR, was achieved only withP. cactorumandP. ramorum. However, while transformations inP. cactorumwere consistent and stable, transformations inP. ramorumwere highly variable and yielded transformants with very weak mycelial growth and abnormal morphology. Our results indicate thatP. cactorumis the best candidate to move forward with CRISPR-Cas9 protocol development and provide insight for future optimization of plasmid transformations in forest Phytophthoras.
Frontal Cortex Hyperactivation and Gamma Desynchrony in Fragile X Syndrome: Correlates of Auditory Hypersensitivity
Authors: Pedapati Ernest , Ethridge Lauren , Liu Yanchen , Liu Rui , Sweeney John , DeStefano Lisa , Miyakoshi Makoto , Razak Khaleel , Schmitt Lauren , Moore David , Gilbert Donald , Wu Steve , Smith Elizabeth , Shaffer Rebecca , Dominick Kelli , Horn Paul , Binder Devin , Erickson Craig
Published: June 2024

Fragile X syndrome (FXS) is an X-linked disorder that often leads to intellectual disability, anxiety, and sensory hypersensitivity. While sound sensitivity (hyperacusis) is a distressing symptom in FXS, its neural basis is not well understood. It is postulated that hyperacusis may stem from temporal lobe hyperexcitability or dysregulation in topdown modulation. Studying the neural mechanisms underlying sound sensitivity in FXS using scalp electroencephalography (EEG) is challenging because the temporal and frontal regions have overlapping neural projections that are difficult to differentiate. To overcome this challenge, we conducted EEG source analysis on a group of 36 individuals with FXS and 39 matched healthy controls. Our goal was to characterize the spatial and temporal properties of the response to an auditory chirp stimulus. Our results showed that males with FXS exhibit excessive activation in the frontal cortex in response to the stimulus onset, which may reflect changes in top-down modulation of auditory processing. Additionally, during the chirp stimulus, individuals with FXS demonstrated a reduction in typical gamma phase synchrony, along with an increase in asynchronous gamma power, across multiple regions, most strongly in temporal cortex. Consistent with these findings, we observed a decrease in the signal-to-noise ratio, estimated by the ratio of synchronous to asynchronous gamma activity, in individuals with FXS. Furthermore, this ratio was highly correlated with performance in an auditory attention task. Compared to controls, males with FXS demonstrated elevated bidirectional frontotemporal information flow at chirp onset. The evidence indicates that both temporal lobe hyperexcitability and disruptions in top-down regulation play a role in auditory sensitivity disturbances in FXS. These findings have the potential to guide the development of therapeutic targets and back-translation strategies.
An analysis of international strategies for monitoring and preventability described in medicinal product information: a study protocol
Authors: Sartori Daniele , Aronson Jeffrey K. , Norén G. Niklas , Onakpoya Igho J.
Published: June 2024

Introduction

Product information is intended to be a reference for healthcare professionals to verify instructions for monitoring and preventability of adverse drug reactions (ADRs), among other things. International comparisons of these documents, using the Systematic Information for Monitoring (SIM) method, have highlighted discrepancies in the instructions for monitoring, but there has been no comparison of preventability instructions.

Objectives

To quantify and compare, across different countries, the completeness of instructions for monitoring and preventability provided to healthcare professionals in medicinal product information.

Methods

We shall retrieve information included with medicinal products that have been involved in signals communicated by regulators, in 2014–2019 and based on clinical assessments of reports of ADRs, from the websites of 35 regulatory agencies. We shall evaluate the completeness of instructions for monitoring using a modified version of the SIM method; a score of 67% will qualify a monitoring instruction as sufficiently complete. To evaluate the completeness of instructions for preventability, we have derived a framework from the Dose-responsiveness-Temporality-Susceptibility (DoTS) classification of ADRs and related implications, comprising domains and items/implications. We shall iteratively develop a threshold to define the sufficiency of completeness of instructions based on data distribution across DoTS domains. We shall present descriptive statistics by country for each item of the framework and by total scores, using tables, or figures where necessary.

Outcomes

Our target audience is regulators, and the results should highlight gaps in the level of information available to healthcare professionals. This study may also provide some insights into how suspicions of causality that arise from clinical assessments of reports of ADRs translate into actionable recommendations in clinical practice.
The Safety and Efficacy of Cangrelor in Endovascular Thrombectomy Compared with Glycoprotein IIb/IIIa Inhibitors
Authors: Devarajan Alex , Gottiparthi Shouri , Caton Michael T. , Ouf Aya , Wu Katty , Goldman Daryl , Davis Nicole , Musallam Nadine , Zhang Jack Y. , Rao Naina , Dangayach Neha , Davy Connor , Fara Michael , Majidi Shahram , Oxley Thomas , Kellner Christopher , Shigematsu Tomoyoshi , DeLeacy Reade , Mocco J , Fifi Johanna T. , Shoirah Hazem
Published: June 2024

Background

Cangrelor, an intravenous P2Y12-receptor inhibitor, is a reversible and short-acting antithrombotic medication non-inferior to irreversible Glycoprotein IIb-IIIa inhibitors (GPI) like eptifibatide. However, there is insufficient data to compare the medications in endovascular thrombectomy (EVT) cases requiring emergent platelet inhibition. This study reviews our experience with cangrelor in EVT and compares its safety and efficacy against GPIs.

Methods

A large healthcare system retrospective review identified all patients who received cangrelor or eptifibatide intraoperatively during EVT from December 2018 to March 2023 for a cohort study. Clinical data was reviewed. Functional status was defined by the modified Rankin Scale (mRS) and National Institutes of Health Stroke Scale (NIHSS) at multiple time points. Binary variables were tested with Pearson χ2 tests or Fisher’s exact tests. Continuous variables were tested with two-tailed t-tests or Wilcoxon tests.

Results

Of 1,010 EVT patients, 36 cangrelor and 104 eptifibatide patients were selected. There were no significant differences in baseline functional status or presentations. Cangrelor was most frequently administered for stenting tandem occlusions (n=16, 44.4%) and successful reperfusion occurred in 93.3% of patients (n=30). On multivariate analysis, cangrelor usage was associated with decreased odds of hemorrhagic conversion (adjusted odds ratio (AOR) 0.76, p=0.004) and symptomatic hemorrhage (AOR 0.86, p=0.021). There were no significant differences in thrombotic re-occlusion. Cangrelor was associated with lower 24-hour NIHSS (7.0 vs. 12.0, p=0.013) and discharge NIHSS scores (3.0 vs 9.0, p=0.009). There were no significant differences in in-hospital mortality or length of stay. Cangrelor was associated with improved odds of favorable outcome, defined as mRS 0-2, at discharge (AOR 2.69, p=0.001) and on 90-day follow-up (AOR 2.23, p=0.031).

Conclusion

Cangrelor was associated with a decreased risk of hemorrhagic conversion and may lead to favorable functional outcomes for patients during hospitalization when compared to GPIs. Future prospective studies are warranted to investigate its use in EVT.

Previous Presentations

This abstract was previous presented as a podium presentation at the Society of Neurointerventional Surgery’s 20thAnnual Meeting in San Diego, CA from July 31st- August 4th, 2023.
Evidence for auditory stimulus-specific adaptation but not deviance detection in larval zebrafish brains
Authors: Wilde Maya , Poulsen Rebecca , Qin Wei , Arnold Joshua , Favre-Bulle Itia A. , Mattingley Jason B , Scott Ethan K , Stednitz Sarah Josephine
Published: June 2024

Animals receive a constant stream of sensory input, and detecting changes in this sensory landscape is critical to their survival. One signature of change detection in humans is the auditory mismatch negativity (MMN), a neural response to unexpected stimuli that deviate from a predictable sequence. This process requires the auditory system to adapt to specific repeated stimuli while remaining sensitive to novel input (stimulus-specific adaptation). MMN was originally described in humans, and equivalent responses have been found in other mammals and birds, but it is not known to what extent this deviance detection circuitry is evolutionarily conserved. Here we present the first evidence for stimulus-specific adaptation in the brain of a teleost fish, using whole-brain calcium imaging of larval zebrafish at single-neuron resolution with selective plane illumination microscopy. We found frequency-specific responses across the brain with variable response amplitudes for frequencies of the same volume, and created a loudness curve to model this effect. We presented an auditory 'oddball' stimulus in an otherwise predictable train of pure tone stimuli, and did not find a population of neurons with specific responses to deviant tones that were not otherwise explained by stimulus-specific adaptation. Further, we observed no deviance responses to an unexpected omission of a sound in a repetitive sequence of white noise bursts. These findings extend the known scope of auditory adaptation and deviance responses across the evolutionary tree, and lay groundwork for future studies to describe the circuitry underlying auditory adaptation at the level of individual neurons.
A novel variant inADPRSdisrupts ARH3 stability and subcellular localization in children with neurodegeneration and respiratory failure
Authors: Bannister Maxwell , Bray Sarah , Aggarwal Anjali , Billington Charles , Nguyen Hai Dang
Published: June 2024

Purpose

ADP-ribosylation is a post-translational modification involving the transfer of one or more ADP-ribose units from NAD+ to target proteins. Dysregulation of ADP-ribosylation is implicated in neurodegenerative diseases. Here we report a novel homozygous variant in theADPRSgene (c.545A>G, p.His182Arg) encoding the mono(ADP-ribosyl) hydrolase ARH3 found in 2 patients with childhood-onset neurodegeneration with stress-induced ataxia and seizures (CONDSIAS).

Methods

Genetic testing via exome sequencing was used to identify the underlying disease cause in two siblings with developmental delay, seizures, progressive muscle weakness, and respiratory failure following an episodic course. Studies in a cell culture model uncover biochemical and cellular consequences of the identified genetic change.

Results

The ARH3H182Rvariant affects a highly conserved residue in the active site of ARH3, leading to protein instability, degradation, and reduced expression. ARH3H182Radditionally fails to localize to the nucleus. The combination of reduced expression and mislocalization of ARH3H182Rresulted in accumulation of mono-ADP ribosylated species in cells.

Conclusions

The children’s clinical course combined with the biochemical characterization of their genetic variant develops our understanding of the pathogenic mechanisms driving CONDSIAS and highlights a critical role for ARH3-regulated ADP ribosylation in nervous system integrity.
Neuronal identity control at the resolution of a single transcription factor isoform
Authors: Smolin Natalie , Dombrovski Mark , Hina Bryce W. , Moreno-Sanchez Anthony , Gossart Ryan , Carmona Catherine R. , Rehan Aadil , Hussein Roni H. , Mirshahidi Parmis , Ausborn Jessica , Kurmangaliyev Yerbol Z. , von Reyn Catherine R.
Published: June 2024

The brain exhibits remarkable neuronal diversity which is critical for its functional integrity. From the sheer number of cell types emerging from extensive transcriptional, morphological, and connectome datasets, the question arises of how the brain is capable of generating so many unique identities. ‘Terminal selectors’ are transcription factors hypothesized to determine the final identity characteristics in post-mitotic cells. Which transcription factors function as terminal selectors and the level of control they exert over different terminal characteristics are not well defined. Here, we establish a novel role for the transcription factorbroadas a terminal selector inDrosophila melanogaster. We capitalize on existing large sequencing and connectomics datasets and employ a comprehensive characterization of terminal characteristics including Perturb-seq and whole-cell electrophysiology. We find a single isoformbroad-z4serves as the switch between the identity of two visual projection neurons LPLC1 and LPLC2.Broad-z4is natively expressed in LPLC1, and is capable of transforming the transcriptome, morphology, and functional connectivity of LPLC2 cells into LPLC1 cells when perturbed. Our comprehensive work establishes a single isoform as the smallest unit underlying an identity switch, which may serve as a conserved strategy replicated across developmental programs.
Targeting the paralog protein for degradation accounts for the opposite roles of NRBP1 and NRBP2 in regulating LINE1 retrotransposition
Authors: Yang Wei , Cong Shaobo , Schwarz Jennifer , Schulze Thilo , Ullrich Sara , Falkenstein Kristina , Ott Denis , Eixmann Pia , Trentino Angelica , Thien Antje , Heidmann Thierry , Schulze Ekkehard , Warscheid Bettina , Baumeister Ralf , Qi Wenjing
Published: June 2024

Gene duplication generates paralogs undergoing diverse fates during evolution and serves as a potent catalyst of biological complexity. Paralogs frequently share redundant functions and may also exhibit antagonistic activities by competing for common interaction partners. Here we show that the gene paralogsNRBP1andNRBP2oppositely regulate LINE1 retrotransposition, via influencing integrity of the LINE1 ribonucleoprotein complex. We demonstrate that the opposite roles of NRBP1 and NRBP2 are not results of a competitive mechanism, but rather due to targeting NRBP1 for proteasome-mediated decay by NRBP2, probably through heterodimer formation. Moreover, our phylogenetic analysis shows that the regulatory function of NRBP2 may be acquired later during evolution, suggesting that evolutionary pressure has favored this functional fine-tuning of NRBP1. In summary, our discovery not only identifies NRBP1/2 as novel LINE1 regulators and implicates their involvement in human pathogenesis, but also provides a novel insight into the regulatory details arising from gene duplication.
The continuous differentiation of multiscale structural gradients from childhood to adolescence correlates with the maturation of cortical morphology and functional specialization
Authors: He Yirong , Zeng Debin , Li Qiongling , Chu Lei , Dong Xiaoxi , Liang Xinyuan , Sun Lianglong , Liao Xuhong , Zhao Tengda , Chen Xiaodan , Lei Tianyuan , Men Weiwei , Wang Yanpei , Wang Daoying , Hu Mingming , Pan Zhiying , Zhang Haibo , Liu Ningyu , Tan Shuping , Gao Jia-Hong , Qin Shaozheng , Tao Sha , Dong Qi , He Yong , Li Shuyu
Published: June 2024

From childhood to adolescence, the structural organization of the human brain undergoes dynamic and regionally heterogeneous changes across multiple scales, from synaptic pruning to the reorganization of large-scale anatomical wiring. However, during this period, the developmental process of multiscale structural architecture, its association with cortical morphological changes, and its role in the maturation of functional organization remain largely unknown. Here, we utilized a longitudinal multimodal imaging dataset including 276 children aged 6 to 14 years to investigate the developmental process of multiscale cortical wiring. We used an in vivo model of cortical wiring that combines features of white matter tractography, cortico-cortical proximity, and microstructural similarity to construct a multiscale brain structural connectome. By employing the gradient mapping method, the gradient space derived from the multiscale structural connectome effectively recapitulated the sensory-association axis and anterior-posterior axis. Our findings revealed a continuous expansion of the multiscale structural gradient space during development, with the principal gradient increasingly distinguishing between primary and transmodal regions. This age-related differentiation coincided with regionally heterogeneous changes in cortical morphology. Furthermore, our study revealed that developmental changes in coupling between multiscale structural and functional connectivity were correlated with functional specialization refinement, as evidenced by changes in the participation coefficient. We also found that the differentiation of the principal multiscale structural gradient was associated with improved cognitive abilities, such as enhanced working memory and attention performance, and potentially supported by molecular processes related to synaptic functions. These findings advance our understanding of the intricate maturation process of brain structural organization and its implications for cognitive performance.
Detection of latent brain states from baseline neural activity in the amygdala
Authors: Aucoin Alexa , Lin Kevin K. , Gothard Katalin M.
Published: June 2024

The amygdala responds to a large variety of socially and emotionally salient environmental and interoceptive stimuli. The context in which these stimuli occur determines their social and emotional significance. In canonical neurophysiological studies, the fast-paced succession of stimuli and events induce phasic changes in neural activity. During inter-trial intervals neural activity is expected to return to a stable and relatively featureless baseline. Context, such as the presence of a social partner, or the similarity of trials in a blocked design, induces brain states that can transcend the fast-paced succession of stimuli and can be recovered from the baseline firing rate of neurons. Indeed, the baseline firing rates of neurons in the amygdala change between blocks of trials of gentle grooming touch, delivered by a trusted social partner, and non-social airflow stimuli, delivered by a computer-controlled air valve. In this experimental paradigm, the presence of the groomer alone was sufficient to induce small but significant changes in baseline firing rates. Here, we examine local field potentials (LFP) recorded during these baseline periods to determine whether context was encoded by network dynamics that emerge in the local field potentials from the activity of large ensembles of neurons. We found that machine learning techniques can reliably decode social vs. non-social context from spectrograms of baseline local field potentials. Notably, decoding accuracy improved significantly with access to broad-band information. No significant differences were detected between the nuclei of the amygdala that receive direct or indirect inputs from areas of the prefrontal cortex known to coordinate flexible, context-dependent behaviors. The lack of nuclear specificity suggests that context-related synaptic inputs arise from a shared source, possibly interoceptive inputs that signal the sympathetic- vs. parasympathetic-dominated states characterizing non-social and social blocks, respectively.