Antiretroviral Therapy Suppresses RNA N6-Methyladenosine Modification in Peripheral Blood Mononuclear Cells from HIV-1-Infected Individuals.

RNA N6-methyladenosine (m6A) modification is important for regulating gene expression and innate immune responses to viral infection. HIV-1 in vitro infection induces a significant increase in m6A modification of cellular RNA; however, whether m6A levels of cellular RNA are affected by HIV-1 replication or by antiretroviral therapy (ART) in infected individuals remains unknown. Using dot blot or enzyme-linked immunosorbent assay, we measured RNA m6A levels of peripheral blood mononuclear cells (PBMCs) from healthy donors or HIV-1-infected individuals with or without ART. Using a reverse transcription-quantitative polymerase chain reaction array, we quantified expression levels of 84 type-I interferon (IFN-I)-responsive genes in PBMCs from some individuals of these three groups. RNA m6A levels in PBMCs from HIV-1 viremic patients (n = 10) were significantly higher (p ≤ .0001) compared with ART-treated individuals (n = 22) or 1.5-fold higher compared with healthy donors (n = 14). However, the increase in RNA m6A levels did not correlate with changes in the expression of 10 m6A-regulatory genes. We found significant upregulation and downregulation in the expression of several IFN-I-responsive genes from HIV-1 viremic patients (n = 4) and ART-treated patients (n = 6) compared with healthy donors (n = 5), respectively. Our results suggest that post-transcriptional m6A modification may contribute to the regulation of IFN-I-responsive gene expression during HIV-1 infection and ART.

Signaling controversy and future therapeutical perspectives of targeting sphingolipid network in cancer immune editing and resistance to tumor necrosis factor-α immunotherapy.

Anticancer immune surveillance and immunotherapies trigger activation of cytotoxic cytokine signaling, including tumor necrosis factor-α (TNF-α) and TNF-related apoptosis-inducing ligand (TRAIL) pathways. The pro-inflammatory cytokine TNF-α may be secreted by stromal cells, tumor-associated macrophages, and by cancer cells, indicating a prominent role in the tumor microenvironment (TME). However, tumors manage to adapt, escape immune surveillance, and ultimately develop resistance to the cytotoxic effects of TNF-α. The mechanisms by which cancer cells evade host immunity is a central topic of current cancer research. Resistance to TNF-α is mediated by diverse molecular mechanisms, such as mutation or downregulation of TNF/TRAIL receptors, as well as activation of anti-apoptotic enzymes and transcription factors. TNF-α signaling is also mediated by sphingosine kinases (SphK1 and SphK2), which are responsible for synthesis of the growth-stimulating phospholipid, sphingosine-1-phosphate (S1P). Multiple studies have demonstrated the crucial role of S1P and its transmembrane receptors (S1PR) in both the regulation of inflammatory responses and progression of cancer. Considering that the SphK/S1P/S1PR axis mediates cancer resistance, this sphingolipid signaling pathway is of mechanistic significance when considering immunotherapy-resistant malignancies. However, the exact mechanism by which sphingolipids contribute to the evasion of immune surveillance and abrogation of TNF-α-induced apoptosis remains largely unclear. This study reviews mechanisms of TNF-α-resistance in cancer cells, with emphasis on the pro-survival and immunomodulatory effects of sphingolipids. Inhibition of SphK/S1P-linked pro-survival branch may facilitate reactivation of the pro-apoptotic TNF superfamily effects, although the role of SphK/S1P inhibitors in the regulation of the TME and lymphocyte trafficking should be thoroughly assessed in future studies.

Functional and antigenic landscape of the Nipah virus receptor binding protein.

Nipah virus recurrently spills over to humans, causing fatal infections. The viral receptor-binding protein (RBP or G) attaches to host receptors and is a major target of neutralizing antibodies. Here we use deep mutational scanning to measure how all amino-acid mutations to the RBP affect cell entry, receptor binding, and escape from neutralizing antibodies. We identify functionally constrained regions of the RBP, including sites involved in oligomerization, along with mutations that differentially modulate RBP binding to its two ephrin receptors. We map escape mutations for six anti-RBP antibodies, and find that few antigenic mutations are present in natural Nipah strains. Our findings offer insights into the potential for functional and antigenic evolution of the RBP that can inform the development of antibody therapies and vaccines.

Protocol to process crosslinking and immunoprecipitation data into annotated binding sites.

Here, we present a protocol for using Skipper, a pipeline designed to process crosslinking and immunoprecipitation (CLIP) data into annotated binding sites. We describe steps for partitioning annotated transcript regions and fitting data to a beta-binomial model to call windows of enriched binding. From raw CLIP data, we detail how users can map reproducible RNA-binding sites to call enriched windows and perform downstream analysis. This protocol supports optional customizations for different use cases. For complete details on the use and execution of this protocol, please refer to Boyle et al.1.

Protein nanoparticle vaccines induce potent neutralizing antibody responses against MERS-CoV.

Middle East respiratory syndrome coronavirus (MERS-CoV) is a zoonotic betacoronavirus that causes severe and often lethal respiratory illness in humans. The MERS-CoV spike (S) protein is the viral fusogen and the target of neutralizing antibodies, and has therefore been the focus of vaccine design efforts. Currently there are no licensed vaccines against MERS-CoV and only a few candidates have advanced to Phase I clinical trials. Here we developed MERS-CoV vaccines utilizing a computationally designed protein nanoparticle platform that has generated safe and immunogenic vaccines against various enveloped viruses, including a licensed vaccine for SARS-CoV-2. Two-component protein nanoparticles displaying MERS-CoV S-derived antigens induced robust neutralizing antibody responses and protected mice against challenge with mouse-adapted MERS-CoV. Electron microscopy polyclonal epitope mapping and serum competition assays revealed the specificities of the dominant antibody responses elicited by immunogens displaying the prefusion-stabilized S-2P trimer, receptor binding domain (RBD), or N-terminal domain (NTD). An RBD nanoparticle vaccine elicited antibodies targeting multiple non-overlapping epitopes in the RBD, whereas anti-NTD antibodies elicited by the S-2P- and NTD-based immunogens converged on a single antigenic site. Our findings demonstrate the potential of two-component nanoparticle vaccine candidates for MERS-CoV and suggest that this platform technology could be broadly applicable to betacoronavirus vaccine development.

The Impact of Experience Versus Decision Aids on Patient Preference Toward Virtual Care.

Introduction: Virtual care utilization has increased in recent years bringing questions of how to best inform patients regarding their use. Decision aids (DAs) are tools created to assist patients in making informed decisions about their health care. This study seeks to determine whether a DA or previous experience could better educate and influence patient's preference on virtual care. Methods: One hundred fifty participants from an orthopedic clinic of a multi-hospital system were divided into three groups. Group 1 (Virtual Care Cohort) had at least one previous virtual care visit and was surveyed with the Telemedicine Satisfaction Questionnaire (TSQ). Group 2 (In-person with Decision Aid) and Group 3 (In-person without Decision Aid) had no virtual care experience. Group 2 received a validated virtual care DA with a knowledge test. Both groups were also administered the TSQ. Results: After the DA, patients improved their score on 3 of 4 virtual care knowledge questions. Each cohort demonstrated a positive perception of virtual care; however, the specific reasons for their favorable views varied. The DA cohort did not show increased preference toward virtual care compared with the non-DA group and only responded significantly higher regarding encounter comfort. Patients with previous experience in virtual care responded most favorably to the majority of survey questions regarding their virtual care preferences when compared with both virtual care naive cohorts. Discussion and Conclusion: We found that patient experience was the most important factor in influencing patient preference toward virtual care. Although the DA increased their virtual care knowledge it did not increase their preference; therefore, efforts should be placed at encouraging patient to experience virtual care.

Defining a systems framework for characterizing physical work demands with wearable sensors.

Measuring the physical demands of work is important in understanding the relationship between exposure to these job demands and their impact on the safety, health, and well-being of working people. However, work is changing and our knowledge of job demands should also evolve in anticipation of these changes. New opportunities exist for noninvasive long-term measures of physical demands through wearable motion sensors, including inertial measurement units, heart rate monitors, and muscle activity monitors. Inertial measurement units combine accelerometers, gyroscopes, and magnetometers to provide continuous measurement of a segment's motion and the ability to estimate orientation in 3-dimensional space. There is a need for a system-thinking perspective on how and when to apply these wearable sensors within the context of research and practice surrounding the measurement of physical job demands. In this paper, a framework is presented for measuring the physical work demands that can guide designers, researchers, and users to integrate and implement these advanced sensor technologies in a way that is relevant to the decision-making needs for physical demand assessment. We (i) present a literature review of the way physical demands are currently being measured, (ii) present a framework that extends the International Classification of Functioning to guide how technology can measure the facets of work, (iii) provide a background on wearable motion sensing, and (iv) define 3 categories of decision-making that influence the questions that we can ask and measures that are needed. By forming questions within these categories at each level of the framework, this approach encourages thinking about the systems-level problems inherent in the workplace and how they manifest at different scales. Applying this framework provides a systems approach to guide study designs and methodological approaches to study how work is changing and how it impacts worker safety, health, and well-being.

Time to pregnancy recognition among users of an FDA-cleared fertility application.

Background: Previous investigations of time-to-pregnancy recognition have analysed data from national surveys and clinics, but this has not been investigated in the context of digital fertility applications. Timely pregnancy recognition can help individuals in health and pregnancy management, reducing maternal and foetal risk and costs, whilst increasing treatment options, availability, and cost. Methods: This dataset contained 23,728 pregnancies (conceived between June 2018 and December 2022) from 20,429 participants using a Food and Drug Administration (FDA) cleared fertility app in the United States. Most participants (with non-missing information) identified as Non-Hispanic White, and one-third reported obtaining a university degree. We used two-tailed Welch's t-test, Mann-Whitney U-test, and two-tailed Z-tests to compare time to pregnancy recognition between those using the app to conceive or contracept. Results: Participants using an app to conceive recognised pregnancy on average at 31.3 days from last menstrual period (LMP) compared to 35.9 days among those using the app to prevent pregnancy. Conclusion: Generalisability is limited, as all participants were using a fertility app and had relatively homogenous sociodemographic characteristics.

People who recognise pregnancy early may benefit, as earlier recognition can reduce costs and risks, and make more treatment options available. In the past, researchers have studied the time it takes for an individual to recognise that they are pregnant by asking them in national surveys or when they attend a clinic. However, with the advent of digital fertility tracking apps, we investigated the time it takes to recognise pregnancy when using such an app. We analysed data from 23,728 pregnancies from 20,429 users of the Natural Cycles app between June 2018 and December 2022. We found that participants using the app to try to get pregnant recognised pregnancy an average of 4.6 days earlier than those using the app to prevent pregnancy.

Event-level risk for negative alcohol consequences in emerging adults: The role of affect, motivation, and context.

OBJECTIVE: Decades of research has found support for the motivational model of alcohol use at the between-person level, yet research on event-level drinking motives is in its nascent stage. Similarly, drinking context has been largely ignored in studies of day-level motives. Therefore, the present study sought to test whether drinking context mediates the relation between affect and motivation on drinking outcomes at both day and person levels. METHOD: Emerging adults who drank in solitary and social settings (N = 107; 61.2% female) completed 21 days ecological momentary assessments. Affect was assessed during morning/afternoon reports; drinking motives were assessed during afternoon reports; and past-night drinking context, drinking quantity, and negative consequences were assessed during next morning reports. Two-level multilevel structural equation models tested whether within-person and between-person levels of predrinking affect were indirectly associated with negative consequences through predrinking motives, drinking context (social vs. solitary), and drinking quantity. RESULTS: At the day and person levels, positive affect was associated with higher social and enhancement motives. At the day level, positive affect indirectly predicted consequences through social motives, social (vs. solitary) drinking, and drinking quantity, whereas positive affect indirectly predicted consequences through enhancement motives and drinking quantity above and beyond context. At the day and person levels, negative affect was associated with coping motives, but coping was not associated with context, drinking quantity, nor consequences. CONCLUSIONS: Findings suggest that positive affect was linked to drinking outcomes through motives (enhancement and social) and contexts (social), whereas negative affect was not. Findings suggest that positively valenced drinking motives may be an important just-in-time intervention target. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

Longitudinal changes in sexual desire and attraction among women who started using the Natural Cycles app.

Many women experience sexual side effects, such as decreased libido, when taking hormonal contraceptives (HCs). However, little is known about the extent to which libido recovers after discontinuing HCs, nor about the timeframe in which recovery is expected to occur. Given that HCs suppress the activities of multiple endogenous hormones that regulate both the ovulatory cycle and women's sexual function, resumption of cycles should predict libido recovery. Here, using a combination of repeated and retrospective measures, we examined changes in sexual desire and partner attraction (among partnered women) across a three-month period in a sample of Natural Cycles users (Survey 1: n = 1596; Survey 2: n = 550) who recently discontinued HCs. We also tested whether changes in these outcomes coincided with resumption of the ovulatory cycle and whether they were associated with additional factors related to HC use (e.g., duration of HC use) or relationship characteristics (e.g., relationship length). Results revealed that both sexual desire and partner attraction, on average, increased across three months after beginning to use Natural Cycles. While the prediction that changes in sexual desire would co-occur with cycle resumption was supported, there was also evidence that libido continued to increase even after cycles resumed. Together, these results offer new insights into relationships between HC discontinuation and women's sexual psychology and lay the groundwork for future research exploring the mechanisms underlying these effects.

Disaggregating within- and between-person associations to test the aversive transmission of alcohol use in late adolescence through adulthood.

OBJECTIVE: The theory of aversive transmission posits that children of parents who have an alcohol use disorder (AUD) may abstain or limit their own alcohol use because they believe themselves to be at risk of developing problems with alcohol. The present study examined relationships among parental AUD, perceived parental AUD, perceived risk for AUD, addiction avoidance reasons for limiting alcohol use, and alcohol use using a random intercept cross-lagged panel model. METHOD: Participants (N = 805; 48% female; 28% Latinx) were from a longitudinal study investigating intergenerational transmission of AUD. Parental AUD, perceived parental AUD, perceived risk for AUD, addiction avoidance reasons for limiting alcohol use, and alcohol use (quantity, frequency, and frequency of heavy drinking) were measured every 5 years from late adolescence (Mage = 20) to adulthood (Mage = 32). Random intercept cross-lagged panel models tested whether there were stable between-person relations or time-varying within-person relations among these variables. RESULTS: At the between-person level, perceived parental AUD predicted greater addiction avoidance reasons for limiting alcohol use and greater perceived risk. Those with greater addiction avoidance reasons for limiting alcohol use were less likely to use any alcohol and drank less frequently. Parental AUD was associated with higher levels of alcohol use as well as perceived risk. No consistent cross-lagged paths were found at the within-person level. CONCLUSIONS: Study findings were at the between-person level rather than the within-person level. Future work on aversive transmission is needed to better understand this subgroup of children of parents with AUD. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

Differentiating action from inaction: Longitudinal relations among impulsive personality traits, internalizing symptoms, and drinking behavior.

BACKGROUND: Impulsive personality traits are strong, consistent risk factors for heavy drinking, and modern theories suggest that impulsive traits may also confer risk for internalizing symptoms. However, it remains unclear which specific impulsive traits are linked with heavy drinking versus internalizing symptoms, and whether heavy drinking and internalizing symptoms are mechanisms of risk for negative alcohol consequences in impulsive individuals. METHOD: Data are from a longitudinal study of young adults (N = 448, Mage = 22.27, 43.5 % female) assessed at baseline (T1), 6 months (T2), and one year later (T3). Longitudinal path models tested whether T1 impulsive traits (i.e., lack of premeditation, lack of perseverance, sensation seeking, positive urgency, negative urgency) were indirectly associated with T3 negative alcohol consequences through heavy T2 drinking and T2 internalizing symptoms (i.e., depression, anxiety, stress). Separate models were tested for positive and negative urgency given strong correlations between these measures. RESULTS: Across models, T1 lack of premeditation indirectly predicted more T3 negative alcohol consequences through heavy T2 drinking. When tested separately, T1 negative urgency indirectly predicted more T3 negative consequences through higher T2 stress and depressive (but not anxiety) symptoms, and T1 positive urgency predicted higher T2 anxiety symptoms, but T2 anxiety was unrelated to T3 negative consequences. Across models, T1 sensation seeking indirectly predicted less T3 negative consequences through decreased T2 depression. CONCLUSIONS: Distinct impulsive traits prospectively predicted heavy drinking and internalizing symptoms, both of which conferred risk for negative alcohol consequences. Findings underscore the importance of targeted interventions based on personality and suggest that decreases in drinking may be more effective prevention for those who lack premeditation, whereas decreases in internalizing, particularly depression/stress, may be critical for those high in negative urgency.

A phage nucleus-associated RNA-binding protein is required for jumbo phage infection.

Large-genome bacteriophages (jumbo phages) of the proposed family Chimalliviridae assemble a nucleus-like compartment bounded by a protein shell that protects the replicating phage genome from host-encoded restriction enzymes and DNA-targeting CRISPR-Cas nucleases. While the nuclear shell provides broad protection against host nucleases, it necessitates transport of mRNA out of the nucleus-like compartment for translation by host ribosomes, and transport of specific proteins into the nucleus-like compartment to support DNA replication and mRNA transcription. Here, we identify a conserved phage nuclear shell-associated protein that we term Chimallin C (ChmC), which adopts a nucleic acid-binding fold, binds RNA with high affinity in vitro, and binds phage mRNAs in infected cells. ChmC also forms phase-separated condensates with RNA in vitro. Targeted knockdown of ChmC using mRNA-targeting dCas13d results in accumulation of phage-encoded mRNAs in the phage nucleus, reduces phage protein production, and compromises virion assembly. Taken together, our data show that the conserved ChmC protein plays crucial roles in the viral life cycle, potentially by facilitating phage mRNA translocation through the nuclear shell to promote protein production and virion development.

Do Subjective Effects from Alcohol and Cannabis Predict Simultaneous Use During a Decision-Making Task?

Background: Simultaneous alcohol and cannabis use is associated with negative outcomes, yet little is known about what motivates the decision of simultaneous use. One possibility is that early-episode subjective effects motivate simultaneous use to complement or replace the first substance's effects. The current study used a hypothetical decision-making task to test this hypothesis. Objectives: College students (N = 486) were presented eight scenarios characterized by alcohol/cannabis subjective effects (i.e., high/low arousal positive [e.g., excited, relaxed], high/low arousal negative [e.g., aggressive, dizzy]) and asked their likelihood of simultaneously using the other substance per scenario. Multilevel modeling tested whether subjective effect scenarios predicted a higher likelihood of simultaneous use and whether ordering moderated this association. Results: Task-based simultaneous use likelihood was associated with self-reported simultaneous use, showing task validity. Scenarios characterized by high/low arousal positive effects were associated with higher likelihood of simultaneous use, whereas high/low arousal negative scenarios were associated with lower likelihood. Alcohol vs. cannabis-first scenarios were associated with higher likelihood of simultaneous use, and significant interactions were observed for high/low arousal positive and high arousal negative effects. High arousal positive scenarios were associated with higher likelihood of simultaneous use when cannabis was used first, low arousal positive scenarios with higher likelihood when alcohol was used first, and high arousal negative scenarios with lower likelihood when cannabis was used first. Conclusions: Beginning-of-episode subjective substance effects may be a promising event-level predictor of simultaneous use, and just-in-time interventions may benefit from targeting the ordering and subjective experiences of alcohol and cannabis use.

Return to Sport After Ulnar Collateral Ligament Tears Treated with Platelet-Rich Plasma Injections is Influenced by Length of Rehabilitation and Leukocyte Content of Injections: A Systematic Review.

PURPOSE: To analyze the current literature assessing return to sport (RTS) outcomes after platelet-rich plasma (PRP) injections for the nonoperative treatment of ulnar collateral ligament (UCL) injuries. METHODS: A systematic review of PubMed, Embase, and Web of Science databases was conducted in June 2023 to identify studies assessing RTS after PRP injections for UCL injuries. Tear severity, leukocyte content of PRP, rehabilitation protocol, and RTS outcomes were collected. Heterogeneity was assessed through proportional random-effects models for RTS and return to preinjury level of play (RTLP) with subgroup analysis by rehabilitation length, leukocyte content of PRP, and tear severity. RESULTS: Eight studies with 278 partial-thickness and 44 full-thickness tears were identified. The mean age of patients ranged from 17.3 to 26 years. The mean RTS time after injection ranged from 5.2 to 25.4 weeks. High heterogeneity was observed among studies, with RTS rates ranging from 46% to 100% (I2 = 83%) and RTLP rates ranging from 34% to 100% (I2 = 83%). Studies with the longest rehabilitation programs (12-14 weeks) had RTS rates of 87% to 100% (I2 = 0%). RTS rates among athletes treated with leukocyte-poor and leukocyte-rich PRP ranged from 73% to 100% (I2 = 30%) and 52% to 88% (I2 = 84%), respectively. Subanalysis of RTS by tear severity demonstrated high variability, with partial-thickness rates ranging from 59% to 100% (I2 = 55%) and full-thickness rates ranging from 27% to 100% (I2 = 63.2%). CONCLUSIONS: Studies assessing RTS after PRP injections are highly heterogeneous; however, current data suggest nonoperative RTS and RTLP rates ranging from 46% to 100% and 34% to 100%, respectively. Studies with at least 12 weeks of rehabilitation and studies using leukocyte-poor PRP demonstrated low heterogeneity and greater RTS rates. Alternatively, high heterogeneity was observed among both partial- and full-thickness tears. LEVEL OF EVIDENCE: Level IV, systematic review of Level III-IV studies.

Too Tired to Think: Within and Between-Person Relations Among Impulsive Traits, Sleep Duration, and Mental Health Outcomes.

Heavier drinking and depression are common mental health concerns in the USA, yet few studies have sought to understand transdiagnostic risk factors for both. Two health-focused risk factors are impulsive personality traits and sleep duration, but research typically separates the two, precluding additive and interactive relations. The current study sought to test a theoretical model where risk conferred from impulsive traits is heightened when individuals have reduced sleep. Public-access data from the National Longitudinal Study on Adolescent to Adult Health (Add Health) were used to test study hypotheses. Participants reported on impulsive traits (i.e., lack of premeditation, sensation seeking), sleep duration, depression, and drinking across three waves spanning adolescence, emerging adulthood, and adulthood. Multilevel models distinguished risk processes at the between- vs. within-person level. At the between-person level, sensation seeking predicted drinking whereas premeditation predicted depression. Additionally, within-person deviations in both traits were associated with drinking, whereas within-person deviations in premeditation were associated with depression. Sleep duration was protective against outcomes at both levels. However, main effects were qualified by interactions at both levels, such that having below average sleep duration heightened the effects of premeditation at the between-person level, whereas within-person decreases in sleep heightened the effects of sensation seeking at the within-person level. Findings support a theoretical model where poor sleep exacerbates risk conferred from impulsive traits. Risk conferred from impulsive traits diverged based upon level of analysis, suggesting that global and just-in-time interventions may benefit from targeting specific impulsive traits as well as sleep.

Engineering Saccharomyces cerevisiae for fast vitamin-independent aerobic growth.

Chemically defined media for cultivation of Saccharomyces cerevisiae strains are commonly supplemented with a mixture of multiple Class-B vitamins, whose omission leads to strongly reduced growth rates. Fast growth without vitamin supplementation is interesting for industrial applications, as it reduces costs and complexity of medium preparation and may decrease susceptibility to contamination by auxotrophic microbes. In this study, suboptimal growth rates of S. cerevisiae CEN.PK113-7D in the absence of pantothenic acid, para-aminobenzoic acid (pABA), pyridoxine, inositol and/or biotin were corrected by single or combined overexpression of ScFMS1, ScABZ1/ScABZ2, ScSNZ1/ScSNO1, ScINO1 and Cyberlindnera fabianii BIO1, respectively. Several strategies were explored to improve growth of S. cerevisiae CEN.PK113-7D in thiamine-free medium. Overexpression of ScTHI4 and/or ScTHI5 enabled thiamine-independent growth at 83% of the maximum specific growth rate of the reference strain in vitamin-supplemented medium. Combined overexpression of seven native S. cerevisiae genes and CfBIO1 enabled a maximum specific growth rate of 0.33 ± 0.01 h-1 in vitamin-free synthetic medium. This growth rate was only 17 % lower than that of a congenic reference strain in vitamin-supplemented medium. Physiological parameters of the engineered vitamin-independent strain in aerobic glucose-limited chemostat cultures (dilution rate 0.10 h-1) grown on vitamin-free synthetic medium were similar to those of similar cultures of the parental strain grown on vitamin-supplemented medium. Transcriptome analysis revealed only few differences in gene expression between these cultures, which primarily involved genes with roles in Class-B vitamin metabolism. These results pave the way for development of fast-growing vitamin-independent industrial strains of S. cerevisiae.

Long interspersed nuclear elements safeguard neural progenitors from precocious differentiation.

Long interspersed nuclear element-1 (L1 or LINE-1) is a highly abundant mobile genetic element in both humans and mice, comprising almost 20% of each genome. L1s are silenced by several mechanisms, as their uncontrolled expression has the potential to induce genomic instability. However, L1s are paradoxically expressed at high levels in differentiating neural progenitor cells. Using in vitro and in vivo techniques to modulate L1 expression, we report that L1s play a critical role in both human and mouse brain development by regulating the rate of neural differentiation in a reverse-transcription-independent manner.

Age-Related RPE changes in Wildtype C57BL/6J Mice between 2 and 32 Months.

Purpose: This study provides a systematic evaluation of age-related changes in RPE cell structure and function using a morphometric approach. We aim to better capture nuanced predictive changes in cell heterogeneity that reflect loss of RPE integrity during normal aging. Using C57BL6/J mice ranging from P60-P730, we sought to evaluate how regional changes in RPE shape reflect incremental losses in RPE cell function with advancing age. We hypothesize that tracking global morphological changes in RPE is predictive of functional defects over time. Methods: We tested three groups of C57BL/6J mice (young: P60-180; Middle-aged: P365-729; aged: 730+) for function and structural defects using electroretinograms, immunofluorescence, and phagocytosis assays. Results: The largest changes in RPE morphology were evident between the young and aged groups, while the middle-aged group exhibited smaller but notable region-specific differences. We observed a 1.9-fold increase in cytoplasmic alpha-catenin expression specifically in the central-medial region of the eye between the young and aged group. There was an 8-fold increase in subretinal, IBA-1-positive immune cell recruitment and a significant decrease in visual function in aged mice compared to young mice. Functional defects in the RPE corroborated by changes in RPE phagocytotic capacity. Conclusions: The marked increase of cytoplasmic alpha-catenin expression and subretinal immune cell deposition, and decreased visual output coincide with regional changes in RPE cell morphometrics when stratified by age. These cumulative changes in the RPE morphology showed predictive regional patterns of stress associated with loss of RPE integrity.