bluuebunny/crossref_metadata_2025_split huggingface dataset:
Negative allosteric modulation of CB1 cannabinoid receptor signaling decreases intravenous morphine self-administration and relapse in mice
Authors: Oliva Idaira , Kazi Fezaan , Cantwell Lucas N. , Thakur Ganesh A. , Crystal Jonathon D. , Hohmann Andrea G. 
Published: January 2024

AbstractThe endocannabinoid system interacts with the reward system to modulate responsiveness to natural reinforcers, as well as drugs of abuse. Previous preclinical studies suggested that direct blockade of CB1 cannabinoid receptors (CB1R) could be leveraged as a potential pharmacological approach to treat substance use disorder, but this strategy failed during clinical trials due to severe psychiatric side effects. Alternative strategies have emerged to circumvent the side effects of direct CB1 binding through the development of allosteric modulators. We hypothesized that pharmacological inhibition of CB1R signaling through negative allosteric modulation (NAM) would reduce the reinforcing properties of morphine and decrease opioid addictive behaviors. By employing i.v. self-administration in mice, we studied the effects of the CB1-biased NAM GAT358 on morphine intake, relapse-like behavior, and motivation to work for morphine infusions. Our data revealed that GAT358 reduced morphine infusion intake during the maintenance phase of morphine self-administration under fixed ratio 1 schedule of reinforcement. GAT358 decreased morphine-seeking behavior after forced abstinence. Moreover, GAT358 dose-dependently decreased the motivation to obtain morphine infusions in a progressive ratio schedule of reinforcement. Strikingly, GAT358 did not affect the motivation to work for food rewards in an identical progressive ratio task, suggesting that the effect of GAT358 in decreasing opioid self-administration is reward specific. Furthermore, GAT58 did not produce motor ataxia in the rota-rod test. Our results suggest that CB1R NAMs reduced the reinforcing properties of morphine and could represent a viable therapeutic route to safely decrease opioid-addicted behaviors.
Gut microbiome carbon and sulfur metabolisms supportSalmonelladuring pathogen infection
Authors: Leleiwi Ikaia , Kokkinias Katherine , Kim Yongseok , Baniasad Maryam , Shaffer Michael , Sabag-Daigle Anice , Daly Rebecca A. , Flynn Rory M. , Wysocki Vicki H. , Ahmer Brian M. M. , Borton Mikayla A. , Wrighton Kelly C. 
Published: January 2024

AbstractSalmonella entericaserovar Typhimurium is a pervasive enteric pathogen and an ongoing global threat to public health. Ecological studies in theSalmonellaimpacted gut remain underrepresented in the literature, discounting the microbiome mediated interactions that may informSalmonellaphysiology during colonization and infection. To understand the microbial ecology ofSalmonellaremodeling of the gut microbiome, here we performed multi-omics approaches on fecal microbial communities from untreated andSalmonella-infected mice. Reconstructed genomes recruited metatranscriptomic and metabolomic data providing a strain-resolved view of the expressed metabolisms of the microbiome duringSalmonellainfection. This data informed possibleSalmonellainteractions with members of the gut microbiome that were previously uncharacterized.Salmonella-induced inflammation significantly reduced the diversity of transcriptionally active members in the gut microbiome, yet increased gene expression was detected for 7 members, withLuxibacterandLigilactobacillusbeing the most active. Metatranscriptomic insights fromSalmonellaand other persistent taxa in the inflamed microbiome further expounded the necessity for oxidative tolerance mechanisms to endure the host inflammatory responses to infection. In the inflamed gut lactate was a key metabolite, with microbiota production and consumption reported amongst transcriptionally active members. We also showed that organic sulfur sources could be converted by gut microbiota to yield inorganic sulfur pools that become oxidized in the inflamed gut, resulting in thiosulfate and tetrathionate that supportsSalmonellarespiration. Advancement of pathobiome understanding beyond inferences from prior amplicon-based approaches can hold promise for infection mitigation, with the active community outlined here offering intriguing organismal and metabolic therapeutic targets.
Efficacy of auxin-inducible protein degradation inC. eleganstissues using different auxins and TIR1-expressing strains
Authors: Sharma Nidhi , Marques Filipe , Kratsios Paschalis 
Published: January 2024

SummaryThe auxin-inducible degradation system has emerged as a powerful tool to deplete proteins of interest in cells and tissues of various model organisms, includingC. elegans2–5. Here, we present a detailed protocol to perform AID-driven spatiotemporal depletion of specific proteins inC. eleganstissues. First, we introduced the AID degron and a fluorescent reporter at two conserved proteins: (a) the transcription factor CFI-1 (human ARID3), which is expressed in the nucleus of multipleC. elegansneurons and head muscle cells6,7, and (b) the broadly expressed translation initiation factor Y47D3A.21 (human DENR) that localizes in the cytoplasm. Second, we provide a step-by-step guide on how to generateC. elegansstrains suitable for AID-mediated protein (CFI-1 and DENR) depletion. Third, we find that the degree of CFI-1 and DENR depletion inC. eleganstissues is comparable upon treatment with either natural auxin (indole-3-acetic acid (IAA) or a water-soluble synthetic auxin analog (K-NAA). Last, we compare the degree of AID-mediated CFI-1 depletion inC. elegansneurons when the transport inhibitor response 1 (TIR1), component of the SCF ubiquitin ligase complex, is provided in neurons or all somatic cells. Altogether, this protocol provides side-by-side comparisons of different auxins and TIR1-expressing lines. Such comparisons may benefit future studies of AID-mediated protein depletion inC. elegans.Graphical abstractImage provided as pdf (together with Figures).HighlightsEfficient protein depletion inC. eleganstissues upon treatment with either natural or synthetic auxins.Pansomatic TIR1 expression leads to efficient depletion of CFI-1 and DENR.Panneuronal TIR1 expression leads to neuron-specific, yet variable CFI-1 depletion.The AID system is compatible with fluorescence microscopy, Western blotting and behavioral assays.
Dietary tryptophan and genetic susceptibility expand gut microbiota that promote systemic autoimmune activation
Authors: Ma Longhuan , Ge Yong , Brown Josephine , Choi Seung-Chul , Elshikha Ahmed , Kanda Nathalie , Terrell Morgan , Six Natalie , Garcia Abigail , Mohamadzadeh Mansour , Silverman Gregg , Morel Laurence 
Published: January 2024

SummaryTryptophan modulates disease activity and the composition of microbiota in the B6.Sle1.Sle2.Sle3(TC) mouse model of lupus. To directly test the effect of tryptophan on the gut microbiome, we transplanted fecal samples from TC and B6 control mice into germ-free or antibiotic-treated non-autoimmune B6 mice that were fed with a high or low tryptophan diet. The recipient mice with TC microbiota and high tryptophan diet had higher levels of immune activation, autoantibody production and intestinal inflammation. A bloom ofRuminococcus gnavus (Rg),a bacterium associated with disease flares in lupus patients, only emerged in the recipients of TC microbiota fed with high tryptophan.Rgdepletion in TC mice decreased autoantibody production and increased the frequency of regulatory T cells. Conversely, TC mice colonized withRgshowed higher autoimmune activation. Overall, these results suggest that the interplay of genetic and tryptophan can influence the pathogenesis of lupus through the gut microbiota.
Injury Severity is a Key Contributor to Coagulation Dysregulation and Fibrinogen Consumption
Authors: Gosselin Andrew R. , Bargoud Christopher G. , Sawalkar Abhishek , Mathew Shane , Toussaint Ashley , Greenen Matthew , Coyle Susette M. , Macor Marie , Krishnan Anandi , Goswami Julie , Hanna Joseph S. , Tutwiler Valerie 
Published: January 2024

ABSTRACTBackgroundTraumatic injury is a leading cause of death for those under the age of 45, with 40% occurring due to hemorrhage. Severe tissue injury and hypoperfusion lead to marked changes in coagulation, thereby preventing formation of a stable blood clot and increasing hemorrhage associated mortality.ObjectivesWe aimed to quantify changes in clot formation and mechanics occurring after traumatic injury and the relationship to coagulation kinetics, and fibrinolysis.MethodsPlasma was isolated from injured patients upon arrival to the emergency department. Coagulation kinetics and mechanics of healthy donors and patient plasma were compared with rheological, turbidimetric and thrombin generation assays. ELISA’s were performed to determine tissue plasminogen activator (tPA) and D-dimer concentration, as fibrinolytic markers.ResultsSixty-three patients were included in the study. The median injury severity score (ISS) was 17, median age was 37.5 years old, and mortality rate was 30%. Rheological, turbidimetric and thrombin generation assays indicated that trauma patients on average, and especially deceased patients, exhibited reduced clot stiffness, increased fibrinolysis and reduced thrombin generation compared to healthy donors. Fibrinogen concentration, clot stiffness, D-dimer and tPA all demonstrated significant direct correlation to increasing ISS. Machine learning algorithms identified and highlighted the importance of clinical factors on determining patient outcomes.ConclusionsViscoelastic and biochemical assays indicate significant contributors and predictors of mortality for improved patient treatment and therapeutic target detection.ESSENTIALSTraumatic injury may lead to alterations in a patient’s ability to form stable blood clotsA study was performed to assess how trauma severity affects coagulation kineticsKey alterations were observed in trauma patients, who exhibit weaker and slower forming clotsPaired with machine learning methods, the results indicate key aspects contributing to mortality
Cardiovascular risk in ANCA-associated vasculitis: monocyte phenotyping reveals distinctive signatures between serological subsets
Authors: Vegting Yosta , Hanford Katie ML , Jongejan Aldo , Gajadin Gayle RS , Versloot Miranda , van der Bom-Baylon Nelly D , Dekker Tamara , Penne E Lars , van der Heijden Joost W , Houben Eline , Bemelman Frederike J , Neele Annette E , Moerland Perry D , Vogt Liffert , Kroon Jeffrey , Hilhorst Marc L 
Published: January 2024

AbstractObjectivesAnti-neutrophil cytoplasmic antibodies (ANCA)-associated vasculitides (AAV) is associated with an increased cardiovascular risk, particularly the myeloperoxidase AAV serotype (MPO-AAV). Distinct alterations in monocyte phenotypes may cause accelerated atherosclerotic disease in AAV.MethodsA cohort including 43 AAV patients and 19 healthy controls were included for downstream analyses. Extensive phenotyping of monocytes and monocyte-derived macrophages was performed using bulk RNA-sequencing and flow cytometry. Anin vitrotransendothelial migration assay reflecting intrinsic adhesive and migratory capacities of monocytes was employed. Subsequent sub-analyses were performed to investigate differences between serological subtypes.ResultsMonocyte subset analysis showed increased classical monocytes during active disease, whereas non-classical monocytes were decreased. RNA-sequencing revealed upregulation of distinct inflammatory pathways and lipid metabolism-related markers in monocytes of active AAV patients. No differences were detected in the intrinsic monocyte adhesion and migration capacity. Monocytes of MPO-AAV patients in remission expressed genes related to inflammation, coagulation, platelet-binding and interferon signalling, whereas the expression of chemokine receptors indicative of acute inflammation and monocyte extravasation (i.e., CCR2 and CCR5) was increased in monocytes of proteinase-3(PR3)-AAV patients. During active disease, PR3-AAV was linked with elevated serum CRP and increased platelet counts compared to MPO-AAV.ConclusionThese findings highlight changes in monocyte subset composition and activation, but not in the intrinsic migration capacity of AAV monocytes. MPO-AAV monocytes are associated with sustained upregulation of inflammatory genes, whereas PR3-AAV monocytes exhibit chemokine receptor upregulation. These molecular changes may play a role in elevating cardiovascular risk as well as in the underlying pathophysiology of AAV.Key messages- Monocytes are activated during active ANCA-associated vasculitis (AAV) and upregulate lipid metabolism-related markers- AAV monocytes have a normal intrinsic adhesion and migration capacity, although overall monocyte migration likely rises by other mechanisms- The two serological subsets MPO-AAV and PR3-AAV exhibit differences in monocyte activation and chemokine receptor expression
Differential and temporally dynamic involvement of primate amygdala nuclei in face animacy and reward information processing
Authors: Kuraoka Koji , Nakamura Kae 
Published: January 2024

AbstractDecision-making is influenced by both expected reward and social factors, such as who offered the outcomes. Thus, although a reward might originally be independent from social factors, the two elements are closely related. However, whether and how they are processed separately or conjointly remains unclear. Here, we show that neurons in distinct sub-nuclei of the amygdala encode expected reward and face animacy, which is a vital aspect of face perception. Although these encoding processes are distinct, they rely on partially shared neuronal circuits with characteristic temporal dynamics.Two male macaque monkeys made saccades under different social and reward contexts, created by presenting facial images with independent attributes: animacy (a monkey or cartoon face) and associated reward (large or small). The stimulus image was presented twice per trial: during the initial stimulus encoding (S1) and before saccades were made (S2). A longer gaze duration for eye region of the monkey versus cartoon images indicated more robust social engagement for realistic faces. During S1, a similar number of lateral nucleus neurons encoded either animacy only with a monkey-image preference, reward only with a large-reward preference, or both. Conversely, neurons in the basal and central nuclei primarily encoded reward, preferring large-versus small-reward associated face images. The reward-dependent modulation was continuous after S1, but was more conspicuous during S1 in the basal nucleus and during both S1 and S2 in the central nucleus. This anatomically- and temporally-specific encoding in the amygdala may underlie the computation and integration of face animacy and reward information.Significance StatementReward and social information are closely related but originally independent, as both influence our decision-making. The amygdala has been associated with both reward and social information coding. However, whether and how they are processed separately or conjointly by individual neurons in the amygdala remains unclear.We found that neurons in the lateral and basal nuclei encoded face animacy, which is an important aspect of social information, and reward, respectively, during sensory processing. Neurons in the central nucleus encoded reward information during the execution phase. This provides new clarity regarding the mechanisms of separate or integrated social and reward information processing within the amygdala.
Structural insights into peptidoglycan hydrolysis by the FtsEX system inEscherichia coliduring cell division
Authors: Li Jianwei , He Yutong , Xu Xin , Alcorlo Martin , Shi Jian , Roper David I. , A. Hermoso Juan , Sham Lok-To , Luo Min 
Published: January 2024

AbstractBacterial cell division relies on precise peptidoglycan (PG) remodelling, a process orchestrated by the FtsEX complex. Comprised of FtsE and FtsX, this complex collaborates with EnvC, a periplasmic lytic enzyme activator, to regulate septal PG hydrolysis by amidases like AmiB. While recent structural investigations, particularly ofPseudomonas aeruginosaFtsEX (PaeFtsEX), have shed light on complex interactions and proposed activation mechanisms, the structural intricacies governing PG degradation by the FtsEX complex and EnvC inEscherichia colicytokinesis remain unexplored. In this study, we present a comprehensive biochemical and structural analysis ofE. coliFtsEX complexes, unveiling a key role for ATP in complex stabilization that extends across bacterial species. Upon EnvC binding, ATPase activity markedly increases. High-resolution structures ofEcoFtsEX, both in the presence and absence of EnvC, reveal a symmetrical conformation ofEcoFtsEX capable of accommodating the inherent asymmetry of EnvC, mediated by flexible loops within the periplasmic domain. Our negative-staining imaging showcases an elongatedEcoFtsEX/EnvC/AmiB complex reminiscent of thePaeFtsEX system. These findings collectively provide intricate insights into the regulation of PG cleavage by FtsEX inE. coli- a pivotal model system used in pilot genetic studies, suggesting a conserved mechanism for precise hydrolase activation in bacteria.
Rapid classification of a novel ALS-causing I149S variant in superoxide dismutase-1
Authors: Shephard VK , Brown ML , Thompson BA , Harpur A , McAlary L 
Published: January 2024

AbstractVariants of the oxygen free radical scavenging enzyme superoxide dismutase-1 (SOD1) are associated with the neurodegenerative disease amyotrophic lateral sclerosis (ALS). These variants occur in roughly 20% of familial ALS cases, and 1% of sporadic ALS cases. Here, we identified a novel SOD1 variant in a patient in their 50s who presented with movement deficiencies and neuropsychiatric features. The variant was heterozygous and resulted in the isoleucine at position 149 being substituted with a serine (I149S).In silicoanalysis predicted the variant to be destabilising to the SOD1 protein structure. Expression of the SOD1I149Svariant with a C-terminal EGFP tag in neuronal-like NSC-34 cells resulted in extensive inclusion formation and reduced cell viability. Immunoblotting revealed that the intramolecular disulphide between Cys57 and Cys146 was fully reduced for SOD1I149S. Furthermore, SOD1I149Swas highly susceptible to proteolytic digestion, suggesting a large degree of instability to the protein fold. Finally, fluorescence correlation spectroscopy and native-PAGE of cell lysates showed that SOD1I149Swas monomeric in solution in comparison to the dimeric SOD1WT. This experimental data was obtained within 3 months and resulted in the rapid re-classification of the variant from a variant of unknown significance to a clinically actionable likely pathogenic variant.
Community first response for cardiac arrest: comparing phased dispatch policies through Monte Carlo simulation
Authors: van den Berg Pieter L. , Henderson Shane G. , Li Hemeng , Dicker Bridget , Jagtenberg Caroline J. 
Published: January 2024

AbstractBackgroundAdvanced Community First Responder (CFR) systems send so-called phased alerts: notifications with built-in time delays. The policy that defines these delays affects response times, CFR workload and the number of redundant CFR arrivals.MethodsWe compare policies by Monte Carlo Simulation, estimating the three metrics above. We bootstrap acceptance probabilities and response delays from 29,307 rows of historical data covering all GoodSAM alerts in New Zealand between 1-12-2017 and 30-11-2020. We simulate distances between the patient and CFRs by assuming that CFRs are located uniformly at random in a 1 km circle around the patient, for different CFR densities. Our simulated CFRs travel with a distance-dependent speed that was estimated by linear regression on observed speeds among those responders in the abovementioned data set that eventually reached the patient.ResultsThe alerting policy has a large impact on the expected number of alerts sent, the redundant arrivals and the probability of patient survival. CFR app managers can use our results to identify a policy that displays a desirable trade-off between these performance measures.
Physiological JNK3 Concentrations Are Higher in Motor-related and Disease-implicated Brain Regions of C57BL6/J Mice
Authors: Godieva Victoria , Sammoura Ferass , Verrier Paz Sebastian , Han Yoonhee , Di Guida Valentina , Rishel Michael J. , Richardson Jason R. , Chambers Jeremy W. 
Published: January 2024

AbstractThe c-Jun N-terminal kinase 3 (JNK3) is a stress-responsive protein kinase primarily expressed in the central nervous system (CNS). JNK3 exhibits nuanced neurological activities, such as roles in behavior, circadian rhythms, and neurotransmission, but JNK3 is also implicated in cell death and neurodegeneration. Despite the critical role of JNK3 in neurophysiology and pathology, its localization in the brain is not fully understood due to a paucity of tools to distinguish JNK3 from other isoforms. While previous functional and histological studies suggest locales for JNK3 in the CNS, a comprehensive and higher resolution of JNK3 distribution and abundance remained elusive. Here, we sought to define the anatomical and cellular distribution of JNK3 in adult mouse brains. Data reveal the highest levels of JNK3 and pJNK3 were found in the cortex and the hippocampus. JNK3 possessed neuron-type selectivity as JNK3 was present in GABAergic, cholinergic, and dopaminergic neurons, but was not detectable in VGLUT-1-positive glutamatergic neurons and astrocytesin vivo. Intriguingly, higher JNK3 signals were found in motor neurons and relevant nuclei in the cortex, basal ganglia, brainstem, and spinal cord. While JNK3 was primarily observed in the cytosol of neurons in the cortex and the hippocampus, JNK3 appeared commonly within the nucleus in the brainstem. These distinctions suggest the potential for significant differences between JNK3 actions in distinct brain regions and cell types. Our results provide a significant improvement over previous reports of JNK3 spatial organization in the adult CNS and support continued investigation of JNK3’s role in neurophysiology and pathophysiology.
Multiple intersecting pathways are involved in the phosphorylation of CPEB1 to activate translation during mouse oocyte meiosis
Authors: Kunitomi Chisato , Romero Mayra , Daldello Enrico Maria , Schindler Karen , Conti Marco 
Published: January 2024

AbstractThe RNA-binding protein cytoplasmic polyadenylation element binding 1 (CPEB1) plays a fundamental role in the regulation of mRNA translation in oocytes. However, the nature of protein kinase cascades modulating the activity of CPEB1 is still a matter of controversy. Using genetic and pharmacological tools and detailed time courses, here we have reevaluated the relationship between CPEB1 phosphorylation and the activation of translation during mouse oocyte maturation. We show that both the CDK1/MAPK and AURKA/PLK1 pathways converge on the phosphorylation of CPEB1 during prometaphase. Only inactivation of the CDK1/MAPK pathway disrupts translation, while inactivation of either pathway leads to CPEB1 stabilization. However, stabilization of CPEB1 induced by inactivation of the AURKA/PLK1 does not affect translation, indicating that destabilization/degradation can be dissociated from translational activation. The accumulation of the endogenous CCNB1 protein closely recapitulates the translation data. These findings support the overarching hypothesis that the activation of translation in prometaphase in mouse oocytes relies on a CDK1-dependent CPEB1 phosphorylation, and this translational activation precedes CPEB1 destabilization.