bluuebunny/crossref_metadata_2025_split huggingface dataset:
Epiboly in zebrafish requires reactive oxygen species produced by NADPH oxidases for the regulation of vesicular trafficking
Authors: Ramírez-Corona Arlen , Reza-Medina Brenda , Schnabel Denhi , Lomeli Hilda , Salas-Vidal Enrique 
Published: December 2024

ABSTRACTEpiboly is the first morphogenetic cell movement that occurs at the onset of gastrulation in zebrafish. During epiboly, the blastoderm thins out and spreads cells over the massive yolk cell. Epiboly progression is controlled by a complex regulatory network that involves diverse molecular effectors. Previously, we reported that reactive oxygen species (ROS) derived from NADPH oxidases (Nox) are required for normal epiboly progression, embryo survival, and early development. We also found that the inhibition of Nox activity during gastrulation downregulates E-cadherin abundance at the enveloping layer (EVL) cell margins. Since the dynamic localization of E-cadherin at the plasma membrane is highly regulated by endocytosis and vesicular trafficking during epiboly, in the present study, we investigated the effects of Nox inhibition and hydrogen peroxide (H2O2) on endocytosis and in the localization of different proteins important for endosomal trafficking in zebrafish embryos. We show that the simultaneous treatment with the Nox inhibitor VAS2870 and the dynamin 2 (Dnm2) inhibitor dynasore rescues the effects of VAS2870 on epiboly delay, embryo mortality and E-cadherin abundance at EVL cell margins. Furthermore, we found that H2O2impacts the endocytic rate of fluorescent fluid-phase markers at the EVL, as well as the localization and abundance of Rab11, a small GTPase protein involved in recycling endosomes. Our results suggest that Nox-derived ROS participate in the regulation of the initial steps of endocytosis and in the endosomal trafficking required for epiboly progression during early zebrafish development.HIGHLIGHTS-NADPH oxidase (Nox) activity is required for the epiboly and localization of E- cadherin.-Dynamin inhibition rescues the developmental defects produced by the loss of Nox activity.-Nox-derived reactive oxygen species (ROS) participate in the regulation of endosome and E-cadherin trafficking, which is required for epiboly.-Nox inhibition increases the rate of fluorescent fluid-phase markers of endocytosis in EVL cells.-H2O2decreases fluid-phase internalization in EVL cells.-H2O2regulates Rab11 localization
A flow cytometric approach to identifying the relative abundance and functional capacities of hemocyte subsets in the American cockroach,Periplaneta americana(L.)
Authors: Boyer-Millander Faith J. , Martin Aaron T. , Hamm Chadwick A. , Appel Arthur G. , Hiltbold Schwartz Elizabeth 
Published: December 2024

AbstractThere is growing interest in insects as subjects for comparative immunological studies; however, very little has been done to wholistically characterize insect immune cells using modern techniques such as flow cytometry, and virtually no work of this kind has been done inPeriplaneta americana. Here, we use an array of general molecular probes including fluorescent lectins, lysosomal indicators, and functional assays to distinguish and characterize insect immune cells (hemocytes) based on cell markers and functions. We have utilized fluorescent tracers of lysosomal content, ROS production, and phagocytosis, as well as microscopic examination of morphology and melanization to distinguish hemocyte types based on these functions. Our findings support the use of lectins as an additional means of separating at least three populations of cockroach immune cells coupled with morphological measurements such as size and complexity. Our results indicate that many functions are enriched in the more granular hemocyte population as these are the cells that undergo phagocytosis and melanization.
Mutation inWdr45leads to early motor dysfunction and widespread aberrant axon terminals in a beta-propeller protein associated neurodegeneration (BPAN) patient-inspired mouse model
Authors: Meyerink Brandon L , Karia Krishna S , Rechtzigel Mitchell J , Patthi Prithvi R , Edwards Ariana C , Howard Jessica M , Aaseng Elizabeth R , Weimer Jill M , Pilaz Louis-Jan 
Published: December 2024

AbstractBeta-propeller Protein Associated Neurodegeneration (BPAN) is a devastating neurodevelopmental and neurodegenerative disease linked to variants inWDR45. Currently, there is no cure or disease altering treatment for this disease. This is, in part, due to a lack of insight into early phenotypes of BPAN progression andWDR45’s role in establishing and maintaining neurological function. Here we generated and characterized a mouse model bearing a c52C>T BPAN patient variant inWdr45.We show this mutation ablates WDR45 protein expression and alters autophagy in the brain. Behavioral analysis of these mice revealed characteristic signs of BPAN including cognitive impairment, hyperactivity, and motor decline. We show these behaviors coincide with widespread neuroinflammation and development of axonal spheroids in multiple neuron subclasses throughout the brain. Several lines of evidence suggest these spheroids arise from axon terminals. Transcriptomic analysis uncovered multiple disrupted pathways in the cortex including genes associated with synapses, neurites, endosomes, endoplasmic reticulum, and ferroptosis. This is supported by accumulation of the iron regulating transferrin receptor 1 (TFRC) and the endoplasmic reticulum resident calreticulin (CALR) in the cortex as these animals age. CALR forms spheroid structures similar to the axonal spheroids seen in these animals. Taken together, our data demonstrate that WDR45 is necessary for healthy brain function and maintenance of axon terminals. This model opens the door to therapeutics targeting BPAN and further exploration of the role of WDR45 in neuronal function.
Quantifying the contribution of somatosensory afferent types and changes therein to pain sensitivity using transcutaneous optogenetic stimulation in behaving mice
Authors: Xie Yu-Feng , Dedek Christopher , Prescott Steven A. 
Published: December 2024

ABSTRACTOptogenetics provides an unprecedented opportunity to delineate how different somatosensory afferents contribute to sensation, including pain. By expressing channelrhodopsin-2 (ChR2) in certain afferents, those afferents can be selectively activated by transcutaneous photostimuli applied to behaving mice. Despite the great care taken to precisely target expression of ChR2, imprecise photostimulation has hindered quantitative behavioral testing. Here, using a robot to reproducibly photostimulate behaving mice and precisely measure their paw withdrawal, we show that selectively activating nociceptors with ramped photostimuli evokes faster withdrawal than co-activating nociceptive and non-nociceptive afferents, consistent with gate control. We also show that inflammation-induced hyperexcitability in nociceptors is sufficient to increase pain sensitivity. Electrophysiological testing confirmed that inflammation increases nociceptor excitability without affecting phototransduction. Data further suggest that withdrawal latency depends on the number of nociceptors activated rather than how strongly each nociceptor is activated. Consistent with changes described in nociceptor somata, the behavioral consequences of peripherally blocking different voltage-gated sodium (NaV) channels showed that nociceptor axons normally rely on NaV1.8 but upregulate NaV1.7 after inflammation, with important clinical implications for drug efficacy. Collectively, these results demonstrate the utility of optogenetic pain testing when reproducibly delivered and strategically designed photostimuli are used.SIGNIFICANCE STATEMENTTranscutaneous optogenetic stimulation was first applied to behaving mice to explore the neural basis for pain over a decade ago. Despite great care taken to control which afferents express optogenetic actuators, the sensitivity of such testing has been hindered by crude photostimulation methods and imprecise response measurement. Here, we demonstrate highly quantitative optogenetic pain testing using robotic stimulation and withdrawal detection. By comparing paw withdrawal to equivalent nociceptor activation with and without activation of non-nociceptive afferents, we demonstrate the antinociceptive effect of the latter input. We also demonstrate increased pain sensitivity due to inflammation-induced hyperexcitability in nociceptors and the associated change in NaVisoform expression. We also show that withdrawal from ramped optogenetic stimulation reflects how many nociceptors are recruited.
Proteomic analysis reveals complex interactions between PERK/ISR and MAPK1 in translational control in a phenotypic toxin model of Parkinson’s disease expressing PINTAC peptides
Authors: Nallur Girish N 
Published: December 2024

AbstractIn a proteomics investigation of a Parkinson’s disease [PD] cell model, a panel of 7 proprietary PINTAC peptides deployed in anin vitrophenotypic screen in SH-SY5Y cells treated with 6-OHDA identified 2 shorter variants of PERK protein kinase. One of the PERK variants, which is SUMOylated, associatesin vitrowith phosphorylated EIF2 alpha and its ubiquitinated isoforms, which also involves another PERK substrate. PINTACs revealed the presence of multiple post-translational modifications [PTMs] of MAPK1 in presence of OHDA. A fragment derived from MAPK1 protein sequence physically associated with multiple proteins from SH-SY5Y cellsin vitro, including some PERK substrates involved in translational control. Most associations were abolished by OHDA treatment, with concomitant appearance of new interactions of the MAPK1 peptide with RSK proteins. Some PINTACs promoted the potentiation of the function of RPS6viaRSK activation induced by OHDA. A model of the phenotypic effects of aggregator and degrader PINTAC peptides is presented. The significance of these findings for novel drug discovery in PD is discussed in the context of target discovery and therapeutic applications of PINTAC peptides or small molecule mimics.
A2A-positive neurons in the nucleus accumbens core regulate effort exertion
Authors: Shoemaker Charles T. , Friedman Alexander D. , Lu Bryan , Kim Jiwon , Ruan Shaolin , Yin Henry H. 
Published: December 2024

AbstractPrevious work has implicated the nucleus accumbens (NAc) in the regulation of effort, defined as the amount of work an animal is willing to perform for a given reward, but little is known about the specific contributions of neuronal populations within the NAc to effort regulation. In this study, we examined the contributions of direct pathway and indirect pathway neurons in the NAc core using an operant effort regulation task, in which the effort requirement is the number of lever presses needed for earning a food reward. Using optogenetics, we manipulated the activity of direct pathway spiny projection neurons (dSPNs, D1+) and indirect pathway SPNs (iSPNs, A2A+). Activating dSPNs reduced lever pressing regardless of the effort requirement, as it elicited gnawing, a competing consummatory behavior. On the other hand, activating iSPNs in the NAc core (but not in the shell) reduced lever pressing in an effort-dependent manner: stimulation-inducted reduction in performance was greater at higher press-to-reward ratio requirements. In contrast, optogenetically inhibiting NAc core iSPN output resulted in increased levels of effort exertion. Our results show that the indirect pathway output from the NAc core can bidirectionally regulate effort exertion.
Cylicodiscus gabunensis(FABACEAE) aqueous stem bark extract mediated silver nanoparticle enhance anti-inflammation on Wistar rats
Authors: Meva Francois Eya’ane , Mponge Pamela Nguadie , Koube Juliette , Nyuyfoni Gildas Fonye , Hzounda Fokou Jean Baptiste , Ntoumba Agnes Antoinette , Yen Beglau Thi Hai , Tako Djimefo Alex Kevin , Djuidje Annie Guilaine , Tchatchouang Geordamie Chimi , Wounang Ariane Laure , Danielle Evouna Madeleine Ines , Chameni Nkouankam Maeva Jenna , Dada Youte Yolaine Pamela , Ndotti Hassana Moussa , Ebanda Kedi Philippe Belle , Bertin Sone Enone , Janiak Christoph 
Published: December 2024

AbstractIntroductionPlants are a source of bioactive ingredient that can play a key role in development of new drugs. In recent years, plant mediated-biological synthesis of nanoparticles has gained importance due to its simplicity, cost effectiveness and eco-friendly nature. To the best of our readings, nanoparticule fromCylicodiscus gabunensisstem bark is still untaped. This work therefore aimed at assessing the anti-inflammatory properties of the silver nanoparticles obtained from the aqueous extract of the stem bark ofCylicodiscus gabunensis(Cg).MethodologyCylicodiscus gabunensisextract was prepared by infusion-followed by the biosynthesis of silver nanoparticles. The synthesis was monitored by color and UV-Vis spectrophotometry. Infrared spectroscopy aimed at revealing the functional groups present at the surface of the nanoparticles. Structural elucidation was done by powder X-ray crystallography, while microstructure and elemental mapping was performed with scanning electron microscopy and energy dispersive X-ray spectroscopy. In vitro anti-inflammatory test was done using the BSA denaturation based test. The acute toxicity was done using the OEDC 425 guideline. carrageenan-induced rat paw oedema model was used to ascertain thein vivoanti-inflammatory effects.ResultsPhytochemical screening of the aqueous extract ofCylicodiscus gabunensisrevealed the presence of polyphenols, flavonoids, alkaloids, coumarins, saponins, triterpenes, steroids, reducing sugar, tannins, and the absence of anthraquinone. The surface Plasmon resonance peak in the UV-Vis spectrum shows absorption spectra between 380 and 550 nm. Stability studies done over time showed that the nanoparticles were stable even after two months of synthesis. IR spectroscopy revealed the presence of O-H, N-H, C≡C, C=C, C-O, and C=O groups. PXRD confirms the formation of silvernanoparticles (AgNPs) while nanograins of various forms were visualized by SEM and EDX. No toxic sign was observed. The maximum inhibitory percentages were 95% at 200 µg/mL and 91% at 400 μg/Kg forin vitroandin vivoanti-inflammatory effects, respectively.ConclusionThis paper spot the light on silver nanoparticle fromCylicodiscus gabunensisaqueous stem bark for their anti-inflammatory effect on paw oedema model.
Rare habitats, rare species, and invasive predators highlight management complexities in the Colorado River system
Authors: Hossack Blake R. , Stemp Kenzi M. , Goldberg Caren S. , Duke Alexandra C. K. , Preston Taryn N. , Arnold Jeff L. , Ray Andrew M. 
Published: December 2024

AbstractLong-term drought caused Lake Powell, a reservoir on the Colorado River (USA), to decline to its lowest elevation in >50 years during 2022–2023, allowing warm water to pass through intakes of Glen Canyon Dam and facilitating invasion by non-native Smallmouth Bass (Micropterus dolomieu). Establishment of bass downstream of the dam could threaten persistence of several native fishes, including two federally listed species. Subsequent detection of larval Smallmouth Bass in a spring-fed slough (river mile -12 slough) connected to the river in Glen Canyon National Recreation Area (NRA) increased urgency to stem further invasion. The National Park Service is evaluating proposed actions to limit effects from non-native predators on native species in the Colorado River, including potentially channelizing the slough. This locally rare, spring-fed waterbody provides habitat for other species, including Western Tiger Salamanders (Ambystoma mavortiumsubsp.) of uncertain origin. We found salamanders from the slough had two distinct mitochondrial DNA haplotypes identical to sequences from nearby Arizona Tiger Salamander (A. m. nebulosum) populations, confirming they are the native genotype. We detected Red-spotted Toads (Anaxyrus punctatus) and Woodhouse’s Toads (A. woodhousii) from three other sites in Glen Canyon NRA and 34 sites in adjacent, downstream Grand Canyon National Park (spanning ∼464 km of river) with environmental DNA and traditional surveys. However, we did not detect salamanders elsewhere, matching prior information that salamanders are rare in the Colorado River corridor below Glen Canyon Dam. Based on this information, we discuss management options for the local population of Arizona Tiger Salamanders.
dsRNAmax: a Multi-Target Chimeric dsRNA Designer for Safe And Effective Crop Protection
Authors: Fletcher Stephen J , Lawrence Jai , Sawyer Anne , Manzie Narelle , Gardiner Donald , Mitter Neena , Brosnan Christopher 
Published: December 2024

ABSTRACTCrop protection is undergoing significant evolution, transitioning toward sustainable approaches that minimise impacts on the environment and human health. Exogenous application of double-stranded RNA (dsRNA) that silences pest or pathogen genes via RNAi (RNA interference) has promise as a safe and effective next generation crop protection platform without the need for genetic modification. However, exogenous dsRNA application at scale presents challenges. Specifically, a single dsRNA sequence needs to balance targeting the standing variation in a target pest or pathogen group against the potential for adverse impacts in a vast array of non-target and beneficial organisms at the application site and broader environment. To address these competing demands, we present dsRNAmax (https://github.com/sfletc/dsRNAmax), a software package that employsk-mer based assembly of chimeric dsRNA sequences to target multiple related RNA sequences, to broaden the target spectrum. The package ensures designed dsRNAs have no defined contiguous sequence homology with any off-target sequences, which can range from single transcriptomes through to metagenome sequence data and beyond. The efficacy of this package is demonstrated by a dsRNAmax-designed dsRNA that inhibits multiple root-knot nematode species but not a non-target nematode species, despite its susceptibility to environmental RNAi and high homology of the target gene.GRAPHICAL ABSTRACT
ChatGPT as a decision-support tool for better self-monitoring of hearing
Authors: Pastucha Małgorzata , Ratuszniak Anna , Ganc Małgorzata , Piłka Edyta , Drohobycka Iryna , Skarżyński Henryk , Jedrzejczak W. Wiktor 
Published: December 2024

AbstractBackgroundThe rapid development of large language model chatbots, such as ChatGPT, has created new possibilities for healthcare support. This study investigates the feasibility of integrating self-monitoring of hearing (via a mobile app) with ChatGPT’s decision-making capabilities to assess whether specialist consultation is required. In particular, the study evaluated how ChatGPT’s accuracy to make a recommendation changed over periods of up to 12 months.MethodsChatGPT-4.0 was tested on a dataset of 1,000 simulated cases, each containing monthly hearing threshold measurements over periods of up to 12 months. Its recommendations were compared to the opinions of 5 experts using percent agreement and Cohen’s Kappa. A multiple-response strategy, selecting the most frequent recommendation from 5 trials, was also analyzed.ResultsChatGPT aligned strongly with the experts’ judgments, with agreement scores ranging from 0.80 to 0.84. Accuracy scores improved to 0.87 when the multiple-query strategy was employed. In those cases where all 5 experts unanimously agreed, ChatGPT achieved a near-perfect agreement score of 0.99. It adapted its decision-making criteria with extended observation periods, seemingly accounting for potential random fluctuations in hearing thresholds.ConclusionsChatGPT has significant potential as a decision-support tool for monitoring hearing, able to match expert recommendations and adapting effectively to time-series data. Existing hearing self-testing apps lack capabilities for tracking and evaluating changes over time; integrating ChatGPT could fill this gap. While not without its limitations, ChatGPT offers a promising complement to self-monitoring. It can enhance decision-making processes and potentially encourage patients to seek clinical expertise when needed.Graphical abstract
Maladaptive response of an endemic California oak to climatic warming is previewed by interannual variation in growth and survival
Authors: Goetz Alexander R. B. , Ochoa Marissa E. , Wright Jessica W. , Sork Victoria L. 
Published: December 2024

AbstractClimate change poses a major threat to long-lived tree populations by shifting environmental conditions away from those to which species are adapted. One of the biggest concerns is that individuals that are adapted to current conditions will become maladapted to new conditions due to a reduction in fitness. If tree populations are already maladapted to their current environments, climate change may put them at even greater risk. Valley oak (Quercus lobata), a foundational California endemic tree species, has already lost much of its range to anthropogenic activities and has been shown to demonstrate patterns of climatic maladaptation. Given the rapid pace of climate change, predicting the future of a species requires quantifying the extent to which contemporary populations are adapted or maladapted to current climates. Here, we tested the extent and variability of maladaptation in valley oak using a ten-year, range-wide provenance study of 3,371 half-sib juvenile trees from 658 maternal lineages. We compared growth rates and multiplicative fitness functions (height * survival; MFF) across families sourced from sites with differing climates relative to the gardens, as well as against annual phenological records. We found evidence that valley oak trees are most adapted to temperatures beyond the coolest limit of the current species range. Within and across years, trees sourced from hotter localities had higher growth and fitness in the gardens, and the relationship was more pronounced in hotter years. Trees showed persistent maladaptation after ten years, suggesting that the role of phenotypic plasticity in immediate environmental response is superseded by underlying genotypes. Finally, trees with earlier leaf phenology (associated with warmer source climates) consistently showed higher growth rates, further demonstrating a maladaptive pattern. Our results suggest that short-term variation in fitness shows immediate short-term response to climate warming, “previewing” a concerning range of species-level responses to increasing temperature.
Human RNase H2 upregulation counteracts oncogene- and chemotherapy-induced replication stress
Authors: Wilkins Rosanna J. , Kannan Abirami , Plass Siobhan A. , Wilson Claire , Kelly Richard D. W. , Tang Claire H. M. , Kotsantis Panagiotis , Reijns Martin A. M. , Kanhere Aditi , Petermann Eva 
Published: December 2024

AbstractRNase H2 is a heterotrimeric endoribonuclease that resolves RNA:DNA hybrids and genome-embedded ribonucleotides, which are implicated in DNA replication stress and cancer development. Protein and/or mRNA levels of individual RNase H2 subunits are elevated in some cancers, but little is known about the mechanisms or consequences of RNase H2 upregulation. We report that RNase H2 subunits are upregulated at the protein level in response to replication stress induced by oncogenes and chemotherapy drugs in human cancer and non-cancer cell lines. We show that inducible overexpression of the RNASEH2B subunit increases levels of the active RNase H2 heterotrimer. While causing only subtle changes to gene expression, RNASEH2B overexpression is unexpectedly associated with increased RNA:DNA hybrid levels. RNASEH2B overexpression prevents further increases in RNA:DNA hybrid levels by camptothecin or hydroxyurea and reduces replication fork stalling in presence of these drugs. Surprisingly, RNase H2 levels do not strongly impact survival of chemotherapy treatments but appear to have more subtle effects on genome instability and innate immune signalling. In contrast, increased RNase H2 levels in presence of oncogenic HRAS limit not only RAS-induced replication fork stalling but also cell death. Our findings shed new light on the functions of RNase H2 and suggest that upregulation of RNase H2 may be an important aspect of replication stress responses in cancer.