Electron microscopic analysis of rotavirus assembly-replication intermediates

AbstractRotaviruses (RVs) replicate their segmented, double-stranded RNA genomes in tandem with early virion assembly. In this study, we sought to gain insight into the ultrastructure of RV assembly-replication intermediates (RIs) using transmission electron microscopy (EM). Specifically, we examined a replicase-competent, subcellular fraction that contains all known RV RIs. Three never-before-seen complexes were visualized in this fraction. Using in vitro reconstitution, we showed that ~15-nm doughnut-shaped proteins in strings were nonstructural protein 2 (NSP2) bound to viral RNA transcripts. Moreover, using immunoaffinity-capture EM, we revealed that ~20-nm pebble-shaped complexes contain the viral RNA polymerase (VP1) and RNA capping enzyme (VP3). Finally, using a gel purification method, we demonstrated that ~30–70-nm electron-dense, particle-shaped complexes represent replicase-competent core RIs, containing VP1, VP3, and NSP2 as well as capsid proteins VP2 and VP6. The results of this study raise new questions about the interactions among viral proteins and RNA during the concerted assembly–replicase process

The Moderating Effect of Comfort from Companion Animals and Social Support on the Relationship between Microaggressions and Mental Health in LGBTQ+ Emerging Adults

Introduction: Sexual and/or gender minority (SGM; e.g., lesbian, transgender, nonbinary, LGBTQ+) individuals are frequently exposed to various forms of minority stress that impact their mental health and wellbeing. Microaggressions, a form of minority stress, are defined as unconscious behaviors or statements directed at members of marginalized groups that reflect a hostile or discriminatory message. Microaggressions have been associated with several detrimental outcomes, such as depression and anxiety. Social support has been found to be an important protective factor for SGM emerging adults. Additionally, relationships with companion animals are an underexplored source of support that may be important for SGM individuals. This study aims to explore whether, and to what extent, social support from humans and comfort from companion animals moderates the relationship between SGM-related microaggressions and depressive and anxiety symptoms. Methods: We partnered with five community organizations to recruit our sample, which consisted of 134 SGM emerging adults between the ages of 18 and 21 (Mage = 19.31). Approximately 98.5% of our sample identified with a sexual minority identity, 49.5% identified with a gender minority identity, and 37.3% identified as a racial/ethnic minority. All participants had lived with a companion animal within the past year, with the majority of participants living with a dog and/or a cat. We conducted eight simple moderation analyses to explore whether, and to what extent, comfort from companion animals and human social support individually moderated the relationship between two forms of microaggressions (i.e., interpersonal, environmental) and anxiety and depressive symptoms. Further, we ran four additive moderation analyses to investigate whether comfort from companion animals and social support from humans moderated the relationship between each form of microaggressions and mental health symptoms, when the other moderator was held constant. Results: The results of our simple moderation analyses indicated that social support moderated the relationship between both forms of microaggressions and depressive symptoms (interpersonal: ΔR2 = 0.03, F(1, 125) = 4.74, ꞵ = -0.17, t(125) = -2.18, p = .03; environmental: ΔR2 = 0.02, F(1, 124) = 3.93, ꞵ = -0.19, t(124) = -1.98, p = .05). Our findings suggest that social support acted as a protective factor, because the relationship between exposure to microaggressions and depressive symptoms was not significant when participants reported high levels of social support. Comfort from companion animals also moderated the relationship between interpersonal microaggressions and depressive symptoms (ΔR2 = 0.03, F(1, 125) = 4.78, ꞵ = 0.18, t(125) = 2.19, p = .03). However, comfort from companion animals seemed to exacerbate the association between interpersonal microaggressions and depressive symptoms, as there was a positive and significant relationship between these two variables when participants reported medium or high levels of comfort from companion animals. The results of the additive moderation analyses found that the relationship between exposure to microaggressions and depressive symptoms was positive and significant when social support was low or medium and comfort from companion animals was high or medium. However, when social support was high, the relationship was no longer significant, regardless of the level of comfort from companion animals. Discussion: Our results suggest that social support from humans may be a key protective factor that buffers the relationship between microaggressions and depressive symptoms. Further, these findings also highlight the need to continue investigating the complex role of relationships with companion animals on mental health outcomes for SGM emerging adults. In particular, longitudinal studies are needed to clarify the direction of these relationships, as we are unable to make causal inferences with this cross-sectional study. The results from this study have important implications for future research in this area and practice with SGM populations.https://scholarscompass.vcu.edu/gradposters/1142/thumbnail.jp

The development and process evaluation of PEER : a camp-based programme for adolescents impacted by cancer

Adolescents impacted by their own or a relative’s cancer diagnosis experience significant psychosocial needs. Residential programmes provide opportunities to address these, yet limited evaluation research and unclear reporting of therapeutic and theoretical underpinnings complicate efforts to understand programme effects. This paper reports the development and process evaluation of PEER, a four-day programme with psychosocial (acceptance and commitment therapy, self-compassion) and recreational components for adolescents impacted by their own or a parent/sibling’s cancer. Staff (N = 51) and adolescents (N = 148, 12–17 years) who attended a PEER programme participated in this evaluation. The evaluation of fidelity included measures of facilitators’ confidence to deliver content, adherence to the programme manual, quality of programme delivery, participants’ engagement, and overall satisfaction. The process evaluation included assessment of quality of life, distress, and process variables (psychological flexibility, mindfulness, self-compassion) at pre-programme, post-programme, and two-month follow-up, as well as qualitative feedback from participants and facilitators. Moderation analyses identified predictors of clinically significant improvement in psychosocial outcomes. The programme was delivered with good fidelity, and participants reported high satisfaction and engagement. Approximately 15–20% of participants experienced clinically-meaningful improvements in distress and quality of life; those who reported higher distress and lower baseline psychological flexibility, mindfulness and self-kindness experienced greater improvements. Qualitative feedback additionally evidenced the value of peer connection and support. The evaluation evidences PEER’s feasibility, acceptability and value for adolescents impacted by cancer, particularly those experiencing greater distress. Its success indicates the potential of the therapeutic approaches used, and for community organisations to develop interventions complementing services offered by healthcare systems

Reliability and validity of the Safe Routes to school parent and student surveys

Abstract Background The purpose of this study is to assess the reliability and validity of the U.S. National Center for Safe Routes to School's in-class student travel tallies and written parent surveys. Over 65,000 tallies and 374,000 parent surveys have been completed, but no published studies have examined their measurement properties. Methods Students and parents from two Charlotte, NC (USA) elementary schools participated. Tallies were conducted on two consecutive days using a hand-raising protocol; on day two students were also asked to recall the previous days' travel. The recall from day two was compared with day one to assess 24-hour test-retest reliability. Convergent validity was assessed by comparing parent-reports of students' travel mode with student-reports of travel mode. Two-week test-retest reliability of the parent survey was assessed by comparing within-parent responses. Reliability and validity were assessed using kappa statistics. Results A total of 542 students participated in the in-class student travel tally reliability assessment and 262 parent-student dyads participated in the validity assessment. Reliability was high for travel to and from school (kappa > 0.8); convergent validity was lower but still high (kappa > 0.75). There were no differences by student grade level. Two-week test-retest reliability of the parent survey (n = 112) ranged from moderate to very high for objective questions on travel mode and travel times (kappa range: 0.62 - 0.97) but was substantially lower for subjective assessments of barriers to walking to school (kappa range: 0.31 - 0.76). Conclusions The student in-class student travel tally exhibited high reliability and validity at all elementary grades. The parent survey had high reliability on questions related to student travel mode, but lower reliability for attitudinal questions identifying barriers to walking to school. Parent survey design should be improved so that responses clearly indicate issues that influence parental decision making in regards to their children's mode of travel to school

Fertilization and Tree Species Influence on Stable Aggregates in Forest Soil

Abstract: Background and objectives: aggregation and structure play key roles in the water-holding capacity and stability of soils and are important for the physical protection and storage of soil carbon (C). Forest soils are an important sink of ecosystem C, though the capacity to store C may be disrupted by the elevated atmospheric deposition of nitrogen (N) and sulfur (S) compounds by dispersion of soil aggregates via acidification or altered microbial activity. Furthermore, dominant tree species and the lability of litter they produce can influence aggregation processes. Materials and methods: we measured water-stable aggregate size distribution and aggregate-associated organic matter (OM) content in soils from two watersheds and beneath four hardwood species at the USDA Forest Service Fernow Experimental Forest in West Virginia, USA, where one watershed has received (NH4)2SO4 fertilizer since 1989 and one is a reference/control of similar stand age. Bulk soil OM, pH, and permanganate oxidizable carbon (POXC) were also measured. Research highlights: fertilized soil exhibited decreased macro-aggregate formation and a greater proportion of smaller micro-aggregates or unassociated clay minerals, particularly in the B-horizon. This shift in aggregation to soil more dominated by the smallest (\u3c53 μm) fraction is associated with both acidification (soil pH) and increased microbially processed C (POXC) in fertilized soil. Intra-aggregate OM was also depleted in the fertilized soil (52% less OM in the 53–2000 μm fractions), most strongly in subsurface B-horizon soil. We also document that tree species can influence soil aggregation, as soil beneath species with more labile litter contained more OM in the micro-aggregate size class (\u3c250 μm), especially in the fertilized watershed, while species with more recalcitrant litter promoted more OM in the macro- aggregate size classes (500–2000 μm) in the reference watershed. Conclusions: long-term fertilization, and likely historic atmospheric deposition, of forest soils has weakened macro-aggregation formation, with implications for soil stability, hydrology, and storage of belowground C

Sire contribution to fertilization failure and early embryo survival in cattle

peer-reviewedDespite passing routine laboratory tests of semen quality, bulls used in artificial insemination (AI) exhibit a significant range in field fertility. The objective of this study was to determine whether subfertility in AI bulls is due to issues of sperm transport to the site of fertilization, fertilization failure, or failure of early embryo or conceptus development. In experiment 1, Holstein-Friesian bulls (3 high fertility, HF, and 3 low fertility, LF) were selected from the national population of AI bulls based on adjusted fertility scores from a minimum of 500 inseminations (HF: +4.37% and LF: −12.7%; mean = 0%). Superovulated beef heifers were blocked based on estimated number of follicles at the time of AI and inseminated with semen from HF or LF bulls (n = 3–4 heifers per bull; total 19 heifers). Following slaughter 7 d later, the number of corpora lutea was counted and the uteri were flushed. Recovered structures (oocytes/embryos) were classified according to developmental stage and stained with 4′,6-diamidino-2-phenylindole to assess number of cells and accessory sperm. Overall recovery rate (total structures recovered/total corpora lutea) was 52.6% and was not different between groups. Mean (± standard error of the mean) number of embryos recovered per recipient was 8.7 ± 5.2 and 9.4 ± 5.5 for HF and LF, respectively. Overall fertilization rate of recovered structures was not different between groups. However, more embryos were at advanced stages of development (all blastocyst stages combined), reflected in a greater mean embryo cell number on d 7 for HF versus LF bulls. Number of accessory sperm was greater for embryos derived from HF than for LF bulls. The aim of experiment 2 was to evaluate the effect of sire fertility on survival of bovine embryos to d 15. Day 7 blastocysts were produced in vitro using semen from the same HF (n = 3) and LF (n = 3) bulls and transferred in groups of 5–10 to synchronized heifers (n = 7 heifers per bull; total 42 heifers). Conceptus recovery rate on d 15 was higher in HF (59.4%,) versus LF (45.0%). Mean length of recovered conceptuses for HF bulls was not affected by fertility status. In conclusion, while differences in field fertility among AI sires used in this study were not reflected in fertilization rate, differences in embryo quality were apparent as early as d 7. These differences likely contributed to the higher proportion of conceptuses surviving to d 15 in HF bulls