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The transcriptional coactivators EP300 and CREBBP are critical regulators of gene expression that share high sequence identity but exhibit nonredundant functions in basal and pathological contexts. Here, we report the development of a bifunctional small molecule, MC-1, capable of selectively degrading EP300 over CREBBP. Using a potent aminopyridine-based inhibitor of the EP300/CREBBP catalytic domain in combination with a VHL ligand, we demonstrate that MC-1 preferentially degrades EP300 in a proteasome-dependent manner. Mechanistic studies reveal that selective degradation cannot be predicted solely by target engagement or ternary complex formation, suggesting additional factors govern paralogue-specific degradation. MC-1 inhibits cell proliferation in a subset of cancer cell lines and provides a new tool to investigate the noncatalytic functions of EP300 and CREBBP. Our findings expand the repertoire of EP300/CREBBP-targeting chemical probes and offer insights into the determinants of selective degradation of highly homologous proteins., Competing Interests: The authors declare no competing financial interest., (© 2024 The Authors. Published by American Chemical Society.)

Transfer RNA (tRNA) modifications are crucial for protein synthesis, but their position-specific physiological roles remain poorly understood. Here we investigate the impact of N4-acetylcytidine (ac 4 C), a highly conserved tRNA modification, using a Thumpd1 knockout mouse model. We find that loss of Thumpd1-dependent tRNA acetylation leads to reduced levels of tRNA Leu , increased ribosome stalling, and activation of eIF2α phosphorylation. Thumpd1 knockout mice exhibit growth defects and sterility. Remarkably, concurrent knockout of Thumpd1 and the stress-sensing kinase Gcn2 causes penetrant postnatal lethality, indicating a critical genetic interaction. Our findings demonstrate that a modification restricted to a single position within type II cytosolic tRNAs can regulate ribosome-mediated stress signaling in mammalian organisms, with implications for our understanding of translation control as well as therapeutic interventions., Competing Interests: DECLARATION OF INTERESTS The authors have no positions or financial interests to declare.

Substance use has been identified by Indigenous populations as contributing to health disparities facing their communities. Rates of trauma exposure and post-traumatic stress disorder are higher in Indigenous, compared to non-Indigenous, populations and have been linked to substance use. Historical trauma is thought to be one mechanism underlying substance use and related disorders. The purpose of the present study is to summarize the current state of the literature focusing on the association between trauma (historical and lived) and substance use among Indigenous populations in the United States and Canada. Databases were systematically searched using the preferred reporting items for systematic reviews and meta-analyses statement. The search strategy initially yielded 4,026 articles. After exclusion of ineligible articles, 63 articles remained for synthesis. Results of the present review provide evidence for a positive link between substance use and both historical trauma (i.e., 86.4% of studies) and lived trauma (i.e., 84.7% of studies). Indigenous participants reported that historical trauma and pain related to loss of cultural identity contributed to substance use in their communities. Indigenous participants also consistently described an association between lived trauma and substance use. Despite heterogeneity among Indigenous communities, findings suggest a significant association between trauma and substance use across many different tribes and settings (e.g., reservation/reserve, rural/urban). Indigenous participants identified healing from trauma and reconnecting with culture as necessary components for reducing substance use and maintaining sobriety. With this, the development and implementation of interventions should partner with Indigenous communities in a manner that promotes and enhances cultural values for healing., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

The human acetyltransferase paralogues EP300 and CREBBP are master regulators of lysine acetylation whose activity has been implicated in various cancers. In the half-decade since the first drug-like inhibitors of these proteins were reported, three unique molecular scaffolds have taken precedent: an indane spiro-oxazolidinedione (A-485), a spiro-hydantoin (iP300w), and an aminopyridine (CPI-1612). Despite increasing use of these molecules to study lysine acetylation, the dearth of data regarding their relative biochemical and biological potencies makes their application as chemical probes a challenge. To address this gap, here we present a comparative study of drug-like EP300/CREBBP acetyltransferase inhibitors. First, we determine the biochemical and biological potencies of A-485, iP300w, and CPI-1612, highlighting the increased potencies of the latter two compounds at physiological acetyl-CoA concentrations. Cellular evaluation shows that inhibition of histone acetylation and cell growth closely aligns with the biochemical potencies of these molecules, consistent with an on-target mechanism. Finally, we demonstrate the utility of comparative pharmacology by using it to investigate the hypothesis that increased CoA synthesis caused by knockout of PANK4 can competitively antagonize the binding of EP300/CREBBP inhibitors and demonstrate proof-of-concept photorelease of a potent inhibitor molecule. Overall, our study demonstrates how knowledge of the relative inhibitor potency can guide the study of EP300/CREBBP-dependent mechanisms and suggests new approaches to target delivery, thus broadening the therapeutic window of these preclinical epigenetic drug candidates.

Chemoproteomic profiling is a powerful approach to define the selectivity of small molecules and endogenous metabolites with the human proteome. In addition to mechanistic studies, proteome specificity profiling also has the potential to identify new scaffolds for biomolecular sensing. Here, we report a chemoproteomics-inspired strategy for selective sensing of acetyl-CoA. First, we use chemoproteomic capture experiments to validate the N-terminal acetyltransferase NAA50 as a protein capable of differentiating acetyl-CoA and CoA. A Nanoluc-NAA50 fusion protein retains this specificity and can be used to generate a bioluminescence resonance energy transfer (BRET) signal in the presence of a CoA-linked fluorophore. This enables the development of a ligand displacement assay in which CoA metabolites are detected via their ability to bind the Nanoluc-NAA50 protein "host" and compete binding of the CoA-linked fluorophore "guest". We demonstrate that the specificity of ligand displacement reflects the molecular recognition of the NAA50 host, while the window of dynamic sensing can be controlled by tuning the binding affinity of the CoA-linked fluorophore guest. Finally, we show that the method's specificity for acetyl-CoA can be harnessed for gain-of-signal optical detection of enzyme activity and quantification of acetyl-CoA from cellular samples. Overall, our studies demonstrate the potential of harnessing insights from chemoproteomics for molecular sensing and provide a foundation for future applications in target engagement and selective metabolite detection.

Objective: The COVID-19 pandemic has led to substantial changes in college student alcohol use. Changes in drinking motives may explain some of these changes in drinking patterns. The purpose of the present study is to explore how drinking motives and alcohol use have changed amongst college students considering the timeframes before and after the onset of COVID-19 pandemic (i.e., March 2020) in the United States. We hypothesized that there would be significant changes in drinking motives after March 2020, which would be significantly related to changes in alcohol use., Methods: Participants for the current study were undergraduate students reporting lifetime alcohol use ( n = 198, M age = 21.3, 66.7% female, 86.4% White) recruited through online advertisements in classes to complete an online survey in April 2020. Participants were asked to report on their drinking motives and alcohol use considering the timeframes before and after the onset of closures and stay-at-home orders during the COVID-19 pandemic (i.e., before and since March 2020)., Results: Paired samples t -tests revealed that endorsement of social ( t [171) = 12.79, p < .001, d = 1.16) and conformity motives significantly decreased ( t [170] = 4.46, p < .001, d = 0.31), while endorsement of coping motives significantly increased ( t [172] = -2.70, p = .008, d = .15) after the onset of COVID-19. Linear regression analyses, controlling for drinking motives before March 2020, revealed that changes in enhancement ( β = -.47, p < .001) and coping motives ( β = -.22, p = .04) were significantly associated with changes in alcohol use quantity., Conclusions: Findings of the present study support the need for interventions to target coping and social drinking to reduce risk for alcohol use.

The adaptation of Salmonella enterica serovar Typhimurium to stress conditions involves expression of genes within the regulon of the alternative sigma factor RpoN (σ 54 ). RpoN-dependent transcription requires an activated bacterial enhancer binding protein (bEBP) that hydrolyzes ATP to remodel the RpoN-holoenzyme-promoter complex for transcription initiation. The bEBP RtcR in S. Typhimurium strain 14028s is activated by genotoxic stress to direct RpoN-dependent expression of the RNA repair operon rsr-yrlBA-rtcBA. The molecular signal for RtcR activation is an oligoribonucleotide with a 3'-terminal 2',3'-cyclic phosphate. We show in S. Typhimurium 14028s that the molecular signal is not a direct product of nucleic acid damage, but signal generation is dependent on a RecA-controlled SOS-response pathway, specifically, induction of prophage Gifsy-1. A genome-wide mutant screen and utilization of Gifsy prophage-cured strains indicated that the nucleoid-associated protein Fis and the Gifsy-1 prophage significantly impact RtcR activation. Directed-deletion analysis and genetic mapping by transduction demonstrated that a three-gene region (STM14&#95;3218-3220) in Gifsy-1, which is variable between S. Typhimurium strains, is required for RtcR activation in strain 14028s and that the absence of STM14&#95;3218-3220 in the Gifsy-1 prophages of S. Typhimurium strains LT2 and 4/74, which renders these strains unable to activate RtcR during genotoxic stress, can be rescued by complementation in cis by the region encompassing STM14&#95;3218-3220. Thus, even though RtcR and the RNA repair operon are highly conserved in Salmonella enterica serovars, RtcR-dependent expression of the RNA repair operon in S. Typhimurium is controlled by a variable region of a prophage present in only some strains. IMPORTANCE The transcriptional activator RtcR and the RNA repair proteins whose expression it regulates, RtcA and RtcB, are widely conserved in Proteobacteria . In Salmonella Typhimurium 14028s, genotoxic stress activates RtcR to direct RpoN-dependent expression of the rsr-yrlBA-rtcBA operon. This work identifies key elements of a RecA-dependent pathway that generates the signal for RtcR activation in strain 14028s. This signaling pathway requires the presence of a specific region within the prophage Gifsy-1, yet this region is absent in most other wild-type Salmonella strains. Thus, we show that the activity of a widely conserved regulatory protein can be controlled by prophages with narrow phylogenetic distributions. This work highlights an underappreciated phenomenon where bacterial physiological functions are altered due to genetic rearrangement of prophages.

Alcohol use disorders (AUDs) are among the most prevalent behavioral and mental health diagnoses. Individuals with an AUD are at increased risk for numerous consequences across their social, health, and psychological functioning. Research suggests that differences may exist in the prevalence and consequences of AUD and in the efficacy of AUD treatment across demographic characteristics (i.e., sex/gender and race/ethnicity). This meta-epidemiologic review examined the inclusion of diverse groups (sex/gender and race/ethnicity) in published randomized controlled trials of nonpharmacological treatments for AUD since 1994, following passage of the National Institutes of Health Revitalization Act of 1993. We systematically searched databases following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement criteria. The initial search strategy yielded 7931 articles. After excluding ineligible articles, 155 were included in the present review for synthesis. Of the articles included in this review, only 57 (36.8%) fully reported on both their sample's sex/gender and racial/ethnic breakdown. Of the total sample, seven articles specifically examined one racial/ethnic group and 32 specifically examined one sex/gender group. Six articles (3.9%) reported no information regarding the racial/ethnic breakdown of their sample and five articles (3.2%) reported no information regarding the sex/gender breakdown of their participants. Only two articles (1.3%) reported on subgroup analyses that examined differences in treatment outcomes by both sex/gender and race/ethnicity, despite guidelines set forth by NIH. Only 46 articles (29.7%) described the failure to include diverse sex/gender or racial/ethnic groups or concerns about the generalizability of study findings given their sample's sex/gender or racial/ethnic composition as methodological limitations. These results indicate that substantial efforts must be put forth by the scientific community to ensure the inclusion, analysis, and reporting of data focused on women/females and members of minoritized racial/ethnic groups., (© 2022 by the Research Society on Alcoholism.)

North American Indigenous youth experience disproportionate rates of racial discrimination as well as consequences associated with alcohol use. Self-compassion has been found to be related to both racial discrimination and alcohol use, separately. However, no work to date has examined the role of self-compassion as a moderator of the links among racial discrimination and alcohol use and alcohol-related problems. First Nation adolescents ( N = 106, M age = 14.6, 50.0% female) from reserve communities in Eastern Canada completed a pencil-and-paper survey regarding their experiences of racial discrimination, self-compassion, alcohol use, and alcohol-related problems. The associations between racial discrimination and both alcohol use and alcohol-related problems were significantly moderated by self-compassion. Simple slopes analysis revealed that the associations between racial discrimination and alcohol use was significant for those with low ( b = 6.03, p = .002) but not high ( b = -0.30, p = .88) levels of self-compassion. Similarly, the association between racial discrimination and alcohol-related problems was significant for those with low ( b = 21.81, p = .001) but not high ( b = 0.64, p = .93) levels of self-compassion. Findings of the present study suggest that low levels of self-compassion may increase risk for alcohol use and experiencing negative alcohol-related consequences in the context of racial discrimination among North American Indigenous adolescents. Future work should examine the utility of interventions targeting self-compassion to examine their effects on responses to racial discrimination and alcohol use.

Plasmodium is a genus of apicomplexan parasites which replicate in the liver before causing malaria. Plasmodium vivax can also persist in the liver as dormant hypnozoites and cause clinical relapse upon activation, but the molecular mechanisms leading to activation have yet to be discovered. In this study, we use high-resolution microscopy to characterize temporal changes of the P. vivax liver stage tubovesicular network (TVN), a parasitophorous vacuole membrane (PVM)-derived network within the host cytosol. We observe extended membrane clusters, tubules, and TVN-derived vesicles present throughout P. vivax liver stage development. Additionally, we demonstrate an unexpected presence of the TVN in hypnozoites and observe some association of this network to host nuclei. We also reveal that the host water and solute channel aquaporin-3 (AQP3) associates with TVN-derived vesicles and extended membrane clusters. AQP3 has been previously shown to localize to the PVM of P. vivax hypnozoites and liver schizonts but has not yet been shown in association to the TVN. Our results highlight host-parasite interactions occur in both dormant and replicating liver stage P. vivax forms and implicate AQP3 function during this time. Together, these findings enhance our understanding of P. vivax liver stage biology through characterization of the TVN with an emphasis on the presence of this network in dormant hypnozoites., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Sylvester, Maher, Posfai, Tran, Crawford, Vantaux, Witkowski, Kyle and Derbyshire.)

The demand for opioid medication to effectively treat pain has contributed to the surging opioid crisis, which is a major source of morbidity and mortality in the U.S. More than 100,000 people begin opioid maintenance treatment (OMT) annually, the standard pharmacotherapy for opioid use disorder (OUD). Although OMT is the standard care for OUD, patients often experience or develop a heightened sensitivity to pain (hyperalgesia) as a result of the opioid medication, and also have high rates of stress, affective, and anxiety-related conditions. These conditions are interactive with other behavioral and environmental correlates of opioid and other substance use disorders including impulsive decision-making (e.g., harmful opioid use associated with increased delay discounting), and a lack of alternative (i.e., substance-free) and social reinforcement. Collectively these complex and multifaceted factors constitute significant predictors of lack of adherence to OMT (and other pharmacotherapies) and relapse. There is an urgent need, therefore, to develop novel adjunctive treatments that preserve the benefits of OMT and various pharmacotherapies, and simultaneously diminish continued pain and hyperalgesia, reduce stress and anxiety-related conditions, target relevant behavioral mechanism such as impulsive choice, and also serve to enhance the value of alternative and substance free activities. Here, we discuss evidence that an environmental manipulation - access to greenspace and nature - could serve as a potential adjunctive treatment to standard pharmacotherapies by targeting multiple biological and behavioral mechanisms that standard pharmacotherapies do not address., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Supportive housing programs are encountering a growing phenomenon: aging residents. This phenomenon has created challenges for supportive housing programs nationally, which were not originally designed to serve seniors. This article presents a case study that demonstrates the success of The Bridge's model, MAGIC, to support residents aging in supportive housing.

The participation of cyclooxygenase (COX) in embryo implantation and parturition has been studied extensively. However, the distinct role of the two COX isoforms in these processes still remains unclear. Using three characterized mouse lines where the Ptgs1 and Ptgs2 genes substitute for one another, this study focused on the reproductive significance of their distinct roles and potential biological substitution. In both non-gravid and gravid uteri, the knock-in COX-2 is expressed constitutively, whereas the knock-in COX-1 is slightly induced in early implantation. The delayed onset of parturition previously found in COX-1 null mice was corrected by COX-2 exchange in COX-2>COX-1 mice, with normal term pregnancy, gestation length and litter size. In contrast, loss of native COX-2 in COX-1>COX-2 mice resulted in severely impaired reproductive functions. Knock-in COX-1 failed to substitute for the loss of COX-2 in COX-1>COX-2 mice during implantation, indicating that COX-1 may be replaced by COX-2, but not vice versa. A panel of prostaglandins detected in uterus and ovary demonstrates that prostaglandin biosynthesis preferentially depends on native COX-1, but not COX-2. More interestingly, preferential compensations by the COX isoforms were sustained despite weak dependency on their role in prostaglandin biosynthesis in the uterus and ovary.

Carboxamidines functionalized with either a spiropyran or fulgimide photoswitch were prepared on multigram scales. The thermal, electrochemical, and photochemical ring isomerizations of these compounds were studied and the results compared with related systems. The photochemical isomerisations were found to be reversible and could be followed by 1H NMR and UV-vis spectroscopy. The spiropyran/merocyanine couple was thermally active and an activation enthalpy of 116 kJ mol-1 was measured for ring-opening. These measurements yielded an enthalpy difference of 25 kJ mol-1 between the open and closed states which is consistent with DFT calculations. DFT calculations predicted a charge transfer to the carboxamidine group upon ring closure in the fulgimide and a charge transfer from the carboxamidine group upon switching the spiropyran to the merocyanine form. This was confirmed experimentally by monitoring the change in the oxidation potential assigned to the carboxamidine group. The potential of these molecules to therefore act as a new class of photoresponsive ligands that can modulate the ligand field of a complex is discussed.

Objective: This study analyzed predictors of post-traumatic stress disorder (PTSD) in civilian trauma victims to assess how peritraumatic dissociation (PD) relates to PTSD symptom development. We examined PD and PTSD symptoms from a prior trauma simultaneously to better understand the extent to which past and current reactions to a trauma can predict the development of PTSD for a current trauma., Methods: Participants (N=48) were recruited from the emergency department (ED) of a large, southeastern hospital and assessed immediately after a trauma and again at 4 weeks and 12 weeks post-trauma. We used both self-report and interviewer-based questionnaires to assess PD and PTSD symptoms for prior and current trauma., Results: A hierarchical linear regression revealed that at 4-week follow up, when controlling for several demographic variables and trauma type, a model including both PD and PTSD symptoms from a prior trauma significantly predicted PTSD outcome (F(47)= 3.70, p=0.00), with PD and prior PTSD symptoms significantly contributing 17% and 9% of variance respectively. At 12 weeks, PTSD symptoms from prior trauma (β=0.094, p=0.538) and PD (β=-0.017, p=0.909) did not account for a significant proportion of the variance in PTSD for the enrolling trauma., Conclusion: Prior and current reactions to trauma are both important factors in predicting the development of PTSD symptoms to a current trauma. The more immediate measurement of PD during presentation to the ED may explain the strength of its relationship to PTSD symptom development. Furthermore, our findings support the use of PTSD symptoms of a past trauma, as opposed to trauma frequency, as a predictor of PTSD from a subsequent trauma. Methodological limitations and future directions are discussed.

Subclavian venous catheterisation, using a Seldinger technique, was attempted in 25 children weighing less than 10 kg (mean 6.4 kg). Catheterisation was successful in 24 cases. Initial catheter position was satisfactory in 19 of the 24 cases (79%) and with repositioning it was possible to obtain satisfactory flow and pressure-wave recordings in all 24 patients. The technique is simple and provides reliable access to the central circulation in small infants. There was no serious morbidity attributable to the technique.

A study of controlled hypotension was undertaken in 50 major surgical patients using labetalol, a drug with both alpha- and beta-adrenoceptor blocking activity. The patients were such that difficulty in achieving controlled hypotension could be anticipated: the young, the anxious and those for whom halothane was contraindicated. The administration of labetalol quickly induced hypotension which was controlled easily and was rapidly antagonized.