Single-cell topographical profiling of the immune synapse reveals a biomechanical signature of cytotoxicity.

Immune cells have intensely physical lifestyles characterized by structural plasticity and force exertion. To investigate whether specific immune functions require stereotyped mechanical outputs, we used super-resolution traction force microscopy to compare the immune synapses formed by cytotoxic T cells with contacts formed by other T cell subsets and by macrophages. T cell synapses were globally compressive, which was fundamentally different from the pulling and pinching associated with macrophage phagocytosis. Spectral decomposition of force exertion patterns from each cell type linked cytotoxicity to compressive strength, local protrusiveness, and the induction of complex, asymmetric topography. These features were validated as cytotoxic drivers by genetic disruption of cytoskeletal regulators, live imaging of synaptic secretion, and in silico analysis of interfacial distortion. Synapse architecture and force exertion were sensitive to target stiffness and size, suggesting that the mechanical potentiation of killing is biophysically adaptive. We conclude that cellular cytotoxicity and, by implication, other effector responses are supported by specialized patterns of efferent force.

Automated Cell Lineage Reconstruction using Label-Free 4D Microscopy.

Here we describe embGAN, a deep learning pipeline that addresses the challenge of automated cell detection and tracking in label-free 3D time lapse imaging. embGAN requires no manual data annotation for training, learns robust detections that exhibits a high degree of scale invariance and generalizes well to images acquired in multiple labs on multiple instruments.

Impact of participation as a peer change agent on peer change agents themselves: a quantitative study of a peer-led, social media-based PrEP promotion intervention.

Despite the adoption of pre-exposure prophylaxis (PrEP) as a crucial HIV intervention, uptake remains suboptimal among men who have sex with men, a sexual minority group, due to barriers like cost and stigma. Peer change agents (PCAs) disseminate PrEP information within their social networks. This study explores the reciprocal effects of an online community-based participatory intervention on PCAs, focusing on their transformed PrEP uptake perceptions-leadership efficacy, social network dynamics, attitudes, perceived benefits and barriers and self-efficacy. Leveraging insights from the PrEP Chicago Study, our research addresses a key gap in community-based participatory interventions for PrEP uptake: the transformative experiences and perception shifts of PCAs involved in these interventions. We engaged 20 men who have sex with men, aged 18-45, as PCAs in a one-group pretest-posttest design intervention, which disseminated PrEP communications within their preferred online networks. We utilized the PrEP Chicago Study's 45 Likert items, tailored to reveal the PCAs' transformative potential. Data on PrEP uptake perceptions, sociodemographics and social media use were captured and analyzed using the Wilcoxon matched-pairs signed-rank test, a nonparametric method. PCAs demonstrated significant changes in their PrEP uptake perceptions, including leadership capacity, social network dynamics, attitudes toward PrEP, perceived benefits, barriers and self-efficacy. Our intervention highlights the reciprocal transformation PCAs undergo when disseminating PrEP information. This study adds a new dimension to community-based PrEP interventions and underscores the need for continued refinement of peer-led strategies to optimize the transformative potential of PCAs.

Oncogenic IDH mutations increase heterochromatin-related replication stress without impacting homologous recombination.

Oncogenic mutations in isocitrate dehydrogenases 1 and 2 (IDH1/2) produce 2-hydroxyglutarate (2HG), which inhibits dioxygenases that modulate chromatin dynamics. The effects of 2HG have been reported to sensitize IDH tumors to poly-(ADP-ribose) polymerase (PARP) inhibitors. However, unlike PARP-inhibitor-sensitive BRCA1/2 tumors, which exhibit impaired homologous recombination, IDH-mutant tumors have a silent mutational profile and lack signatures associated with impaired homologous recombination. Instead, 2HG-producing IDH mutations lead to a heterochromatin-dependent slowing of DNA replication accompanied by increased replication stress and DNA double-strand breaks. This replicative stress manifests as replication fork slowing, but the breaks are repaired without a significant increase in mutation burden. Faithful resolution of replicative stress in IDH-mutant cells is dependent on poly-(ADP-ribosylation). Treatment with PARP inhibitors increases DNA replication but results in incomplete DNA repair. These findings demonstrate a role for PARP in the replication of heterochromatin and further validate PARP as a therapeutic target in IDH-mutant tumors.

Re-envisioning the role of student health centers in offering LGBTQIA + friendly and sex-positive services.

Objective: Offering comprehensive lesbian, gay, bisexual, transgender, queer/questioning, intersex, asexual (LGBTQIA+) friendly and sex-positive student health centers are central to institutions of higher education being able to retain and support students. Positive sexual experiences for LGBTQIA + students can have an impact on academic success but many LGBTQIA + students are discriminated against and are often victims of microaggressions. Participants: Twenty undergraduate students enrolled in a LGBTQIA+ health course. Methods: Students in an LGBTQIA + Health undergraduate course at a private Northeastern university assessed what mattered most to them when it came to creating a sex-positive healthcare environment that promotes LGBTQIA + inclusiveness and removes perceived barriers. Results: The themes included expanding mental health care, greater involvement of culturally-competent provider, establishing a stigma-free clinical environment, re-imagining the clinic waiting room, and facilitating sexual health advocacy. Conclusion: Re-envisioning the role of student health centers is critical for ensuring every student has a fair and just opportunity to achieve their full health potential.

Multiplex Spatial Protein Detection by Combining Immunofluorescence with Immunohistochemistry.

Technologies for staining and imaging multiple antigens in single tissue sections are developing rapidly due to their potential to uncover spatial relationships between proteins with cellular resolution. Detections are performed simultaneously or sequentially depending on the approach. However, several technologies can detect limited numbers of antigens or require expensive equipment and reagents. Another serious concern is the lack of flexibility. Most commercialized reagents are validated for defined antibody panels, and introducing any changes is laborious and costly. In this chapter, we describe a method where we combine, for the first time, multiplexed IF followed by sequential immunohistochemistry (IHC) with AEC chromogen on Leica Bond staining processors with paraffin tissue sections. We present data for successful detection of 10 antigens in a single tissue section with preserved tissue integrity. Our method is designed for use with any combination of antibodies of interest, with images collected using whole slide scanners. We include an image viewing and image analysis workflow using nonlinear warping to combine all staining passes in a single full-resolution image of the entire tissue section, aligned at the single cell level.

Trends in nasopharyngeal cancer mortality in the United States, 1999-2020.

OBJECTIVES: The incidence of nasopharyngeal cancer (NPC) has been declining in the United States (US) in recent years. However, little is known about the latest trends in NPC mortality in the US population. This study aimed to examine the trends in NPC mortality rate by age, sex, race and ethnicity and US Census Region from 1999 to 2020. METHODS: Mortality data were extracted from the Centers for Disease Control and Prevention's Wide-ranging Online Data for Epidemiologic Research (WONDER) database. Decedents whose cause of death was NPC were identified using the International Classification of Diseases Codes, 10th Revision: C11.0-C11.9. Trends in age adjusted mortality rates (AAMR) from NPC were assessed using a joinpoint regression model. Annual Percentage Changes (APC) and Average Annual Percentage Changes were examined overall and by age, sex, race and ethnicity and census region. RESULTS: From 1999 through 2020, a total of 14 534 NPC deaths were recorded in the US (AAMR = 0.2 per 100 000; 95% CI: 0.2, 0.2). Overall trends remained stationary throughout the study period. Since 2006, recent trends declined by 6.1% per year (95% CI: -8.4, -3.7) among Non-Hispanic Whites, and by 2.7% per year among Non-Hispanic Blacks, Asians/Pacific Islanders and Hispanics. Trends either stabilized or declined by sex, age and US Census Region. Similar results were obtained when the analysis was restricted to decedents aged 65 years and above. CONCLUSIONS: Stationary or declining trends in NPC mortality could be due to the falling incidence of the disease and/or advances in medical diagnosis and treatment. Considering the enigmatic nature of NPC, future studies should explore the genetic and sociodemographic factors associated with the trends reported in this study.

In vivo tumor immune microenvironment phenotypes correlate with inflammation and vasculature to predict immunotherapy response.

Response to immunotherapies can be variable and unpredictable. Pathology-based phenotyping of tumors into 'hot' and 'cold' is static, relying solely on T-cell infiltration in single-time single-site biopsies, resulting in suboptimal treatment response prediction. Dynamic vascular events (tumor angiogenesis, leukocyte trafficking) within tumor immune microenvironment (TiME) also influence anti-tumor immunity and treatment response. Here, we report dynamic cellular-level TiME phenotyping in vivo that combines inflammation profiles with vascular features through non-invasive reflectance confocal microscopic imaging. In skin cancer patients, we demonstrate three main TiME phenotypes that correlate with gene and protein expression, and response to toll-like receptor agonist immune-therapy. Notably, phenotypes with high inflammation associate with immunostimulatory signatures and those with high vasculature with angiogenic and endothelial anergy signatures. Moreover, phenotypes with high inflammation and low vasculature demonstrate the best treatment response. This non-invasive in vivo phenotyping approach integrating dynamic vasculature with inflammation serves as a reliable predictor of response to topical immune-therapy in patients.

Knowledge, Attitudes, and Practices towards PrEP from Cisgender Men and Transgender Women Who Have Sex with Men in the Largest Suburban HIV Epidemic.

We conducted a cross-sectional web-based study to assess attitudes and experiences with HIV pre-exposure prophylaxis (PrEP) amongst a multiracial cohort of gay, bisexual, and other men who have sex with men (MSM) and transgender women in Long Island, New York. Participants were recruited through clinical providers and community-based organizations. The survey assessed knowledge and attitudes toward PrEP and factors that facilitate willingness to take it. Of the 189 respondents, most participants were Latinx/Hispanic (57.1%; n = 105), gay-identifying (81.2%; n = 151), and cisgender men (88.7%; n = 165). One in five participants completed high school or lower (19.4%; n = 36). Among those who had never used PrEP (53.4%; n = 101), nearly all participants were willing to use it if it were free or covered as part of their insurance (89.4%; n = 84). The most common barriers to not using PrEP was not knowing where to obtain it (68.3%; n = 69), concerns about side effects (42.1%; n = 35), and concerns about affordability (38.5%; n = 25). This study discusses specific nuances to the suburbs, including cultural norms and structural barriers that should be incorporated in health promotion initiatives in addressing these factors.

Cross-modality synthesis of EM time series and live fluorescence imaging.

Analyses across imaging modalities allow the integration of complementary spatiotemporal information about brain development, structure, and function. However, systematic atlasing across modalities is limited by challenges to effective image alignment. We combine highly spatially resolved electron microscopy (EM) and highly temporally resolved time-lapse fluorescence microscopy (FM) to examine the emergence of a complex nervous system in Caenorhabditis elegans embryogenesis. We generate an EM time series at four classic developmental stages and create a landmark-based co-optimization algorithm for cross-modality image alignment, which handles developmental heterochrony among datasets to achieve accurate single-cell level alignment. Synthesis based on the EM series and time-lapse FM series carrying different cell-specific markers reveals critical dynamic behaviors across scales of identifiable individual cells in the emergence of the primary neuropil, the nerve ring, as well as a major sensory organ, the amphid. Our study paves the way for systematic cross-modality data synthesis in C. elegans and demonstrates a powerful approach that may be applied broadly.

Effect of modifiable risk factors in Parkinson's disease: A case-control study looking at common dietary factors, toxicants, and anti-inflammatory medications.

OBJECTIVE: To investigate how common modifiable exposures, including dietary factors, select toxicants, and anti-inflammatory medications, may affect Parkinson's disease. METHODS: Using surveys, a case-control study was conducted at a medical center, comparing Parkinson's disease patients (N = 149) and healthy controls (N = 105). Subjects reported exposure to red meats, vegetables, alcohol, tobacco, anti-inflammatory medications, and pesticides. The relationship between exposures and Parkinson's disease diagnosis was analyzed by logistic regression to generate odds ratio and 95% confidence interval. RESULTS: Consuming red meat "sometimes" or "always" was positively associated with Parkinson's disease as compared to eating red meats "rarely" or "never"; (odds ratio = 2.15, 95% confidence interval = 1.06, 4.39; p = 0.03) and (odds ratio = 4.47, 95% confidence interval = 1.67, 11.94; p = 0.003), respectively. Exposure to pesticides showed a positive association with Parkinson's disease (odds ratio = 2.84, 95% confidence interval = 1.34, 6.00; p = 0.007). "Always" use of aspirin was inversely associated with Parkinson's disease (odds ratio = 0.32, 95% confidence interval = 0.14, 0.70; p = 0.004). "Ever" having used anti-histamines was inversely associated with Parkinson's disease (odds ratio = 0.37, 95% confidence interval = 0.17, 0.81; p = 0.01). DISCUSSION: Our study suggests that there are modifiable external factors that are associated with Parkinson's disease. The present study can thus assist clinicians, policy makers, and people living with Parkinson's disease in improving the experience and management of Parkinson's disease.

Differential adhesion regulates neurite placement via a retrograde zippering mechanism.

During development, neurites and synapses segregate into specific neighborhoods or layers within nerve bundles. The developmental programs guiding placement of neurites in specific layers, and hence their incorporation into specific circuits, are not well understood. We implement novel imaging methods and quantitative models to document the embryonic development of the C. elegans brain neuropil, and discover that differential adhesion mechanisms control precise placement of single neurites onto specific layers. Differential adhesion is orchestrated via developmentally regulated expression of the IgCAM SYG-1, and its partner ligand SYG-2. Changes in SYG-1 expression across neuropil layers result in changes in adhesive forces, which sort SYG-2-expressing neurons. Sorting to layers occurs, not via outgrowth from the neurite tip, but via an alternate mechanism of retrograde zippering, involving interactions between neurite shafts. Our study indicates that biophysical principles from differential adhesion govern neurite placement and synaptic specificity in vivo in developing neuropil bundles.

HIV testing behaviors and willingness to receive oral rapid HIV testing among dental patients in Xi'an, China.

INTRODUCTION: HIV testing is an important strategy for controlling and ultimately ending the global pandemic. Oral rapid HIV testing (ORHT) is an evidence-based strategy and the evidence-based shows is favored over traditional blood tests in many key populations. The dental setting has been found to be a trusted, convenient, and yet untapped venue to conduct ORHT. This study assessed the HIV testing behaviors and willingness to receive ORHT among dental patients in Xi'an, China. METHODS: A cross-sectional survey of dental patients from Xi'an was conducted from August to September 2017. Dental patients were recruited using a stratified cluster sampling. A 44-item survey was used to measure HIV/AIDS knowledge, HIV testing behaviors, and willingness to receive ORHT. RESULTS: Nine hundred and nine dental patients completed the survey with a mean HIV/AIDS knowledge score of 10.7/15 (SD 2.8). Eighty-four participants (9.2%) had previously received an HIV test. Participants would have a high rate of HIV testing if they had higher monthly income (OR = 1.982, 95% CI: 1.251-3.140) and a higher HIV/AIDS knowledge score (OR = 1.137, 95% CI: 1.032-1.252). Five hundred and eighty-two participants (64.0%) were willing to receive ORHT before a dental treatment, 198 (21.8%) were not sure, and 129 (14.2%) were unwilling. Logistic regression showed that age (OR = 0.970, 95% CI: 0.959-0.982), HIV/AIDS knowledge score (OR = 1.087, 95% CI: 1.031-1.145), previous HIV test (OR = 2.057, 95% CI: 1.136-3.723), having advanced HIV testing knowledge (OR = 1.570, 95% CI: 1.158-2.128), and having advanced ORHT knowledge (OR = 2.074, 95%: CI 1.469-2.928) were the factors affecting the willingness to receive ORHT. CONCLUSIONS: The majority of dental patients had not previously received an HIV test, although many were receptive to being tested in the dental setting. The dental setting as a venue to screen people for HIV needs further exploration, particularly because many people do not associate dentistry with chairside screenings. Increasing awareness of ORHT and reducing testing price can further improve the patient's willingness to receive ORHT.

The role of mask mandates, stay at home orders and school closure in curbing the COVID-19 pandemic prior to vaccination.

BACKGROUND: COVID-19 has quickly spread throughout the world, necessitating assessment of effective containment methods. The purpose of this study was to examine the impact of government mandated school closures, stay at home orders and mask requirements METHODS: Cumulative incidence rates were calculated at 14-day intervals until the day of the first vaccine administration in the country. Rate ratios were calculated using negative binomial regression while investigating the effects of adjusting for several sociodemographic and medical factors RESULTS: Faster implementation of mask mandates was consistently shown to be protective. States with mask mandates made at three to six months had a 1.61 times higher rate than those who implemented within one month (adjusted rate ratio = 1.61, 95% confidence interval: 1.23-2.10, P = .001). States with mask mandates made after 6 months or with no mandate had a 2.16 times higher rate than those who implemented within 1 month (adjusted rate ratio = 2.16, 95% confidence interval: 1.64-2.88, P < .0001). In contrast, both stay at home orders and school closures had no significant influence on disease trajectory. DISCUSSION: The benefits of mask mandates are apparent, especially when mandates were issued within a month. The impact of school closing and stay at home orders were less clear. CONCLUSIONS: Our results suggest that of the different physical distancing measures implemented by the government, mask mandates are the most important.

Structural and developmental principles of neuropil assembly in C. elegans.

Neuropil is a fundamental form of tissue organization within the brain1, in which densely packed neurons synaptically interconnect into precise circuit architecture2,3. However, the structural and developmental principles that govern this nanoscale precision remain largely unknown4,5. Here we use an iterative data coarse-graining algorithm termed 'diffusion condensation'6 to identify nested circuit structures within the Caenorhabditis elegans neuropil, which is known as the nerve ring. We show that the nerve ring neuropil is largely organized into four strata that are composed of related behavioural circuits. The stratified architecture of the neuropil is a geometrical representation of the functional segregation of sensory information and motor outputs, with specific sensory organs and muscle quadrants mapping onto particular neuropil strata. We identify groups of neurons with unique morphologies that integrate information across strata and that create neural structures that cage the strata within the nerve ring. We use high resolution light-sheet microscopy7,8 coupled with lineage-tracing and cell-tracking algorithms9,10 to resolve the developmental sequence and reveal principles of cell position, migration and outgrowth that guide stratified neuropil organization. Our results uncover conserved structural design principles that underlie the architecture and function of the nerve ring neuropil, and reveal a temporal progression of outgrowth-based on pioneer neurons-that guides the hierarchical development of the layered neuropil. Our findings provide a systematic blueprint for using structural and developmental approaches to understand neuropil organization within the brain.

Dental Care Utilization of Hospitalized Persons Living with HIV and Substance Use.

People living with HIV (PLWH) who use drugs experience worse health outcomes than their non-using counterparts. Little is known about how often they seek dental care and the factors that influence their utilization. PLWH with substance use disorders who were inpatients at 11 urban hospitals (n = 801) participated in a National Institute on Drug Abuse Clinical Trials Network study to improve engagement in HIV outcomes. Dental care utilization at each time point during the study period (baseline, 6 months and/or 12 months) was assessed (n = 657). Univariate analysis and logistic regression were used to examine factors associated with dental care utilization. Over half (59.4%) reported not having received any dental care at any timepoint. Participants with less than high school education had lower odds of reporting dental care utilization than those with more than education (aOR = 0.60 [95% CI 0.37-0.99], p = 0.0382). Participants without health insurance also had lower odds of reporting dental care utilization than those with insurance (aOR = 0.50 [95% CI 0.331-0.76], p = 0.0012). Higher food insecurity was associated with having recent dental care utilization (OR = 1.03 [95% CI 1.00, 1.05], p = 0.0359). Additionally, those from Southern states were less likely to report dental care utilization (aOR = 0.55 [95% CI 0.38, 0.79], p = 0.0013). Having health insurance and education are key factors associated with use of dental care for PLWH with substance use disorders. The association between food insecurity and dental care utilization among this population suggests the need for further exploration.

Cadherin preserves cohesion across involuting tissues during C. elegans neurulation.

The internalization of the central nervous system, termed neurulation in vertebrates, is a critical step in embryogenesis. Open questions remain regarding how force propels coordinated tissue movement during the process, and little is known as to how internalization happens in invertebrates. We show that in C. elegans morphogenesis, apical constriction in the retracting pharynx drives involution of the adjacent neuroectoderm. HMR-1/cadherin mediates this process via inter-tissue attachment, as well as cohesion within the neuroectoderm. Our results demonstrate that HMR-1 is capable of mediating embryo-wide reorganization driven by a centrally located force generator, and indicate a non-canonical use of cadherin on the basal side of an epithelium that may apply to vertebrate neurulation. Additionally, we highlight shared morphology and gene expression in tissues driving involution, which suggests that neuroectoderm involution in C. elegans is potentially homologous with vertebrate neurulation and thus may help elucidate the evolutionary origin of the brain.

Spatial mapping of the collagen distribution in human and mouse tissues by force volume atomic force microscopy.

Changes in the elastic properties of living tissues during normal development and in pathological processes are often due to modifications of the collagen component of the extracellular matrix at various length scales. Force volume AFM can precisely capture the mechanical properties of biological samples with force sensitivity and spatial resolution. The integration of AFM data with data of the molecular composition contributes to understanding the interplay between tissue biochemistry, organization and function. The detection of micrometer-size, heterogeneous domains at different elastic moduli in tissue sections by AFM has remained elusive so far, due to the lack of correlations with histological, optical and biochemical assessments. In this work, force volume AFM is used to identify collagen-enriched domains, naturally present in human and mouse tissues, by their elastic modulus. Collagen identification is obtained in a robust way and affordable timescales, through an optimal design of the sample preparation method and AFM parameters for faster scan with micrometer resolution. The choice of a separate reference sample stained for collagen allows correlating elastic modulus with collagen amount and position with high statistical significance. The proposed preparation method ensures safe handling of the tissue sections guarantees the preservation of their micromechanical characteristics over time and makes it much easier to perform correlation experiments with different biomarkers independently.

The role of patient and public involvement in oral health and HIV/AIDS research, practice and policy.

Patient and public involvement (PPI) is a process whereby patients, caregivers, service users and other relevant stakeholders, including the general public, are actively involved and engaged in activities to develop research. The dental research agenda has traditionally been driven by clinicians, where patients and the public have participated in research as subjects; patient and public involvement can contribute to the research agenda including the design and conduct of research by providing unique perspectives gained through lived experience. This panel of the 8th World Workshop on Oral Health and Diseases in AIDS considered the role of people living with HIV (PLHIV) to contribute to oral health and HIV research and policy through a process of involvement and empowerment. The panel introduced the concepts of PPI, described the purpose of PPI, reflected upon the logistic and ethical considerations thereof and considered how PPI had been utilised effectively in HIV research and policy change. The audience discussion focused on ways in which PPI could more readily and consistently be encouraged within oral health research involving PLHIV.

HIV testing: What, where and how?

Globally, one in four people living with HIV are unaware of their status. Current HIV testing services' strategies are falling short of reaching all, and thus, HIV testing should be offered in more decentralized and non-traditional settings such as the dental and community settings. The workshop titled "HIV Testing: What, Where, and How?" provided an overview of concepts and advances in HIV point of care and self-testing diagnostics and a discussion on the implementation of HIV self-test interventions in different healthcare settings and its impact thereof. We also described how to build layperson capacity to deliver HIV testing in community settings. Additionally, we discussed what we have learned from expanding HIV testing beyond the specialist setting. We also noted considerations (i.e. provider willingness, test selection, training and preparing testing environment) dentists and other non-specialist providers need to account for if they are planning to conduct HIV testing. Finally, we highlighted facilitators and barriers to implementing HIV testing in the dental setting on a global scale. These considerations are critical to meeting the UNAIDS 90-90-90 target to help end the HIV/AIDS epidemic.