Local chromatin context regulates the genetic requirements of the heterochromatin spreading reaction.

Heterochromatin spreading, the expansion of repressive chromatin structure from sequence-specific nucleation sites, is critical for stable gene silencing. Spreading re-establishes gene-poor constitutive heterochromatin across cell cycles but can also invade gene-rich euchromatin de novo to steer cell fate decisions. How chromatin context (i.e. euchromatic, heterochromatic) or different nucleation pathways influence heterochromatin spreading remains poorly understood. Previously, we developed a single-cell sensor in fission yeast that can separately record heterochromatic gene silencing at nucleation sequences and distal sites. Here we couple our quantitative assay to a genetic screen to identify genes encoding nuclear factors linked to the regulation of heterochromatin nucleation and the distal spreading of gene silencing. We find that mechanisms underlying gene silencing distal to a nucleation site differ by chromatin context. For example, Clr6 histone deacetylase complexes containing the Fkh2 transcription factor are specifically required for heterochromatin spreading at constitutive sites. Fkh2 recruits Clr6 to nucleation-distal chromatin sites in such contexts. In addition, we find that a number of chromatin remodeling complexes antagonize nucleation-distal gene silencing. Our results separate the regulation of heterochromatic gene silencing at nucleation versus distal sites and show that it is controlled by context-dependent mechanisms. The results of our genetic analysis constitute a broad community resource that will support further analysis of the mechanisms underlying the spread of epigenetic silencing along chromatin.

Integration of clinical pharmacists to assist with medication surveillance for patients receiving gender-affirming hormone therapy in a community ambulatory setting within the United States.

BACKGROUND: Gender-affirming care (GAC) for gender diverse individuals (includes transgender and nonbinary persons) requires a comprehensive approach to medication surveillance, including monitoring and follow-up. Limited access to these health services can present a barrier to follow-up for routine care. Integration of a pharmacist into therapeutic management has shown positive clinical outcomes; however, their involvement with gender-affirming hormone therapy (GAHT), including routine laboratory monitoring, is not well established. OBJECTIVE: This study aimed to describe the development and implementation of a protocol involving the integration of clinical pharmacists into GAC for gender diverse patients in a community ambulatory setting. PRACTICE DESCRIPTION: Cleveland Clinic's Center for LGBTQ+ Care is embedded in a primary care practice and has an established protocol for GAHT management. The health system also has an established model that uses pharmacists for the management of patients, within the primary care and specialty clinic settings, under a collaborative practice agreement (CPA). PRACTICE INNOVATION: The medical director of the Cleveland Clinic's Center for LGBTQ+ Care and the institution's primary care pharmacists collaborated to propose an update to the CPA to include GAHT monitoring to improve access to routine GAHT follow-up. EVALUATION METHODS: GAHT management was approved to be added to the pharmacist CPA in May of 2022 and the pharmacists started seeing patients in February of 2023. The team opted to start with those patients already established on GAHT for at least 6 months, at least 1 year after transition, and with a primary care provider managing their GAHT. CONCLUSION: Access to follow-up for individuals receiving GAHT may be improved through the utilization of pharmacist services. The incorporation of a CPA with pharmacists for GAHT management can allow for a multidisciplinary approach once a patient is on a stable regimen, thereby increasing provider access to new patients.

On the relationship between green space and civic engagement: The roles of well-being, outgroup trust, and activity level.

Much research has been devoted to the positive effect of green space on prosociality, but little is known about its impact on civic engagement. It is also unclear how the effect takes place. This research fills the voids by regressing 2440 US citizen's civic engagement on the vegetation density and park area in their neighborhoods. It further examines if the effect is due to changes in well-being, interpersonal trust, or activity level. Park area predicts higher civic engagement, which is mediated by higher trust in outgroups. However, the data is inconclusive about the effect of vegetation density and the well-being mechanism. In contrast to the activity hypothesis, the effect of parks on civic engagement is stronger in unsafe neighborhoods, suggesting that they are valuable resources to combat neighborhood problems. The results bring insights into how individuals and communities can benefit the most from green spaces in the neighborhood.

Robust humoral and cellular immune responses and low risk for reinfection at least 8 months following asymptomatic to mild COVID-19.

BACKGROUND: Emerging data support detectable immune responses for months after severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and vaccination, but it is not yet established to what degree and for how long protection against reinfection lasts. METHODS: We investigated SARS-CoV-2-specific humoral and cellular immune responses more than 8 months post-asymptomatic, mild and severe infection in a cohort of 1884 healthcare workers (HCW) and 51 hospitalized COVID-19 patients. Possible protection against SARS-CoV-2 reinfection was analyzed by a weekly 3-month polymerase chain reaction (PCR) screening of 252 HCW that had seroconverted 7 months prior to start of screening and 48 HCW that had remained seronegative at multiple time points. RESULTS: All COVID-19 patients and 96% (355/370) of HCW who were anti-spike IgG positive at inclusion remained anti-spike IgG positive at the 8-month follow-up. Circulating SARS-CoV-2-specific memory T cell responses were detected in 88% (45/51) of COVID-19 patients and in 63% (233/370) of seropositive HCW. The cumulative incidence of PCR-confirmed SARS-CoV-2 infection was 1% (3/252) among anti-spike IgG positive HCW (0.13 cases per 100 weeks at risk) compared to 23% (11/48) among anti-spike IgG negative HCW (2.78 cases per 100 weeks at risk), resulting in a protective effect of 95.2% (95% CI 81.9%-99.1%). CONCLUSIONS: The vast majority of anti-spike IgG positive individuals remain anti-spike IgG positive for at least 8 months regardless of initial COVID-19 disease severity. The presence of anti-spike IgG antibodies is associated with a substantially reduced risk of reinfection up to 9 months following asymptomatic to mild COVID-19.

Circulating Markers of Neutrophil Extracellular Traps Are of Prognostic Value in Patients With COVID-19.

OBJECTIVE: The full spectrum of coronavirus disease 2019 (COVID-19) infection ranges from asymptomatic to acute respiratory distress syndrome, characterized by hyperinflammation and thrombotic microangiopathy. The pathogenic mechanisms are poorly understood, but emerging evidence suggest that excessive neutrophil extracellular trap (NET) formation plays a key role in COVID-19 disease progression. Here, we evaluate if circulating markers of NETs are associated with COVID-19 disease severity and clinical outcome, as well as to markers of inflammation and in vivo coagulation and fibrinolysis. Approach and Results: One hundred six patients with COVID-19 with moderate to severe disease were enrolled shortly after hospital admission and followed for 4 months. Acute and convalescent plasma samples as well as plasma samples from 30 healthy individuals were assessed for markers of NET formation: citrullinated histone H3, cell-free DNA, NE (neutrophil elastase). We found that all plasma levels of NET markers were elevated in patients with COVID-19 relative to healthy controls, that they were associated with respiratory support requirement and short-term mortality, and declined to those found in healthy individuals 4 months post-infection. The levels of the NET markers also correlated with white blood cells, neutrophils, inflammatory cytokines, and C-reactive protein, as well as to markers of in vivo coagulation, fibrinolysis, and endothelial damage. CONCLUSIONS: Our findings suggest a role of NETs in COVID-19 disease progression, implicating their contribution to an immunothrombotic state. Further, we observed an association between circulating markers of NET formation and clinical outcome, demonstrating a potential role of NET markers in clinical decision-making, as well as for NETs as targets for novel therapeutic interventions in COVID-19.

Optimal cutting temperature medium embedding and cryostat sectioning are valid for cardiac myofilament function assessment.

To maximize data obtainment from valuable cardiac tissue, we hypothesized that myocardium fixed in optimal cutting temperature (OCT) medium for histology could also be used to investigate the function of myofilament proteins in situ. We compared tissue prepared via conventional liquid nitrogen (LN) snap freezing with tissue fixed in OCT and then sectioned in fiber-parallel orientation. We found that actin-myosin Ca2+ sensitivity, activation rate by Ca2+, cooperativity along the thin filament, as well as cross-bridge cycling rate were unaffected by OCT storage and could reliably be interpreted after sectioning. Absolute values in maximum force generation per cross-sectional area, as well as passive strain, are difficult to investigate after sectioning, as myofibrillar continuity along the preparation cannot be guaranteed. We have shown that myocardial tissue stored in OCT and sectioned before analysis is available for functional analysis, a valuable means of maximizing usage of precious cardiac biopsies.NEW & NOTEWORTHY Myocardial tissue in optimal cutting temperature (OCT) fixation and cryostat sectioning was tested as a means of storing and preparing tissue for myofilament function analysis in relation to conventional liquid nitrogen freezing and dissection. Actomyosin interaction, Ca2+ force activation, and passive compliance were tested. The study concluded that OCT storage and cryostat sectioning do not interfere with the actomyosin cross-bridge dynamics or Ca2+ activation but that absolute tension values suffer and may not be investigated by this method.

Low- and High-Drag Intermittencies in Turbulent Channel Flows.

Recent direct numerical simulations (DNS) and experiments in turbulent channel flow have found intermittent low- and high-drag events in Newtonian fluid flows, at Reτ=uτh/ν between 70 and 100, where uτ, h and ν are the friction velocity, channel half-height and kinematic viscosity, respectively. These intervals of low-drag and high-drag have been termed "hibernating" and "hyperactive", respectively, and in this paper, a further investigation of these intermittent events is conducted using experimental and numerical techniques. For experiments, simultaneous measurements of wall shear stress and velocity are carried out in a channel flow facility using hot-film anemometry (HFA) and laser Doppler velocimetry (LDV), respectively, for Reτ between 70 and 250. For numerical simulations, DNS of a channel flow is performed in an extended domain at Reτ = 70 and 85. These intermittent events are selected by carrying out conditional sampling of the wall shear stress data based on a combined threshold magnitude and time-duration criteria. The use of three different scalings (so-called outer, inner and mixed) for the time-duration criterion for the conditional events is explored. It is found that if the time-duration criterion is kept constant in inner units, the frequency of occurrence of these conditional events remain insensitive to Reynolds number. There exists an exponential distribution of frequency of occurrence of the conditional events with respect to their duration, implying a potentially memoryless process. An explanation for the presence of a spike (or dip) in the ensemble-averaged wall shear stress data before and after the low-drag (or high-drag) events is investigated. During the low-drag events, the conditionally-averaged streamwise velocities get closer to Virk's maximum drag reduction (MDR) asymptote, near the wall, for all Reynolds numbers studied. Reynolds shear stress (RSS) characteristics during these conditional events are investigated for Reτ = 70 and 85. Except very close to the wall, the conditionally-averaged RSS is higher than the time-averaged value during the low-drag events.

3D printing with 2D colloids: designing rheology protocols to predict 'printability' of soft-materials.

Additive manufacturing (AM) techniques and so-called 2D materials have undergone an explosive growth in the past decade. The former opens multiple possibilities in the manufacturing of multifunctional complex structures, and the latter on a wide range of applications from energy to water purification. Extrusion-based 3D printing, also known as Direct Ink Writing (DIW), robocasting, and often simply 3D printing, provides a unique approach to introduce advanced and high-added-value materials with limited availability into lab-scale manufacturing. On the other hand, 2D colloids of graphene oxide (GO) exhibit a fascinating rheology and can aid the processing of different materials to develop 'printable' formulations. This work provides an in-depth rheological study of GO suspensions with a wide range of behaviours from Newtonian-like to viscoelastic 'printable' soft solids. The combination of extensional and shear rheology reveals the network formation process as GO concentration increases from <0.1 vol% to 3 vol%. Our results also demonstrate that the quantification of 'printability' can be based on three rheology parameters: the stiffness of the network via the storage modulus (G'), the solid-to-liquid transition or flow stress (σf), and the flow transition index, which relates the flow and yield stresses (FTI = σf/σy).

The effects of environmental resource and security on aggressive behavior.

Exposure to different environments has been reported to change aggressive behavior, but previous research did not consider the underlying elements that caused such an effect. Based on previous work on environmental perception, we examined the role of environmental resource and security in altering aggression level. In three experiments, participants were exposed to environments that varied in resource (High vs. Low) and security (High vs. Low) levels, after which aggression was measured. The environments were presented through visual priming (Experiments 1-2) and a first-person gameplay (Experiment 3). We observed a consistent resource-security interaction effect on aggression, operationalized as the level of noise blast (Experiment 1) and number of unpleasant pictures (Experiments 2-3) delivered to strangers by the participants. High resource levels associated with higher aggression in insecure conditions, but lower aggression in secure conditions. The findings suggest that the adaptive value of aggression varies under different environmental constraints. Implications are discussed in terms of the effects of adverse environments on aggression, and the nature's effects on social behavior. Aggr. Behav. 43:304-314, 2017. © 2016 Wiley Periodicals, Inc.

Effects of treating gender dysphoria and anorexia nervosa in a transgender adolescent: Lessons learned.

Patients with gender dysphoria and patients with eating disorders often experience discontent with their bodies. Several reports have recognized the co-occurrence of these two conditions, typically in adults who identify as transgender females and desire a more feminine physique. This case report, in contrast, describes a 16-year-old patient with a female sex assigned at birth who first presented with features consistent with anorexia nervosa and later revealed underlying gender dysphoria with a drive for a less feminine body shape. We discuss both the path to recognizing gender dysphoria in this patient as well as the impact of treatment on his eating disorder and overall well-being. This case is one of only a few reports describing a female-to-male transgender patient with an eating disorder and is the first to explore the effects of hormone and surgical intervention in an adolescent patient.