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.

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.

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.

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.

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 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.

HIV testing in the dental setting: A global perspective of feasibility and acceptability.

HIV is no longer a death sentence with early diagnosis and effective treatment contributing to a good prognosis. Surveillance data, however, suggest that one in four people globally living with HIV are unaware of their HIV infection and remains at risk of transmitting their infection if having unprotected sex. A wider array of testing opportunities and settings are thus needed to address this issue of public health significance. Evidence suggests there is a role for the dental team, particularly dentists, in offering chairside point-of-care HIV screening to patients during their dental appointments. Dentists, dental hygienists, and other members of the dental team may still have a role in helping to identify new cases of HIV with the goal of improving health outcomes, addressing health inequalities and improving the quality and quantity of life. This paper reviews the global epidemiology of HIV/AIDS, summarizes the point-of-care HIV testing process, highlights key findings of international studies (from high-, middle-, and low-income countries) on the role of dental professionals in point-of-care HIV testing, and offers suggestions for what additional evidence is needed to make point-of-care HIV testing a routine part of dental care.

Effect of early highly active antiretroviral therapy on viral suppression among newly diagnosed men who have sex with men living with human immunodeficiency virus in Xi'an, China.

BACKGROUND: The number of men who have sex with men (MSM) living with human immunodeficiency virus (HIV) in China has increased rapidly and thus immediate highly active antiretroviral therapy (HAART) after diagnosis was implemented as a strategy to reduce the HIV transmission. METHODS: MSM who were diagnosed with HIV and received HAART between 2013 to 2015 in Xi'an were divided into three groups (>350, 200-350, and <200 cell/µL) according to their baseline CD4+ T cell count. The time of follow-up was calculated from the first date of receiving HAART to December 31, 2016. The CD4+ T cell count was detected with 1 week before or after HAART. The plasma viral loads were tested after 1, 2, and 3 years of treatment. RESULTS: Of 1442 subjects who received HAART, 690 (47.9%) cases were in >350 cell/µL group, whereas 400 (27.7%) cases and 352 (24.4%) cases were in the 200-350 cell/µL group and <200 cell/µL group, respectively. After 1 year of treatment, the viral suppression rate in the <200 cell/µL group was 91.1%, which was significantly lower than the other two groups. The logistic regression results show that the >350 cell/µL group and 200-350 cell/µL group predicted higher viral suppression rates. CONCLUSIONS: Baseline CD4+ T cell count more than 350 cell/µL can improve viral suppression among MSM living with HIV. Furthermore, to reduce the transmission risk, the treatment compliance of people living with HIV with high CD4+ T cell levels should be improved, and their diagnosis to the treatment time should be decreased.

Oral Rapid HIV Testing: Implementation experiences of dental hygiene faculty and students.

Purpose: The goal of oral rapid HIV testing (ORHT) in the dental setting is to identify persons who are unaware of their positive HIV status. The purpose of this study was to describe the experiences of dental hygiene faculty and students who implemented ORHT in university-based dental hygiene clinics and to assess the facilitators and barriers to implementation of ORHT in the dental setting.Methods: Data were collected via semi-structured interviews with dental hygiene faculty and students who conducted ORHT in three dental clinics located in academic institutions. All interview sessions were audio-recorded and transcribed. An inductive approach informed by grounded theory methodology was used to code data and inform theme development. The interview sessions were completed when conceptual saturation was reached.Results: Five themes were identified by the study participants consisting of dental hygiene faculty (n= 8) and dental hygiene students (n=14). Participants felt dental hygienists are qualified to administer ORHT, which fits within their scope of practice; dental hygienists have the skills to feel comfortable offering ORHT without judgement; training is needed with ORHT administration, reading/discussing test results, and counseling for those who receive reactive results; most patients were receptive to being offered the ORHT; and patients accepted the ORHT because it was free, quick to administer and receive results, and convenient since they were already in the dental setting.Conclusion: Results from this study indicate that dental hygienists can play a key role in public health efforts to identify persons who are unaware of their HIV status.

Oral rapid HIV testing in the dental setting: Experiences from three dental hygiene clinics.

Background: Oral rapid HIV testing (ORHT) is implemented in the dental setting to make individuals aware of their possibly undiagnosed HIV infection. The testing process and characteristics of clients willing to receive ORHT has yet, however, to be systematically collected. Case description: Three dental hygiene clinics located in academic institutions implemented ORHT from March 2016 to April 2017. Results: 231 persons received ORHT; all had non-reactive results. Most had seen a primary care provider in the past year (n = 130), had had a previous ORHT (n = 111), and described themselves as extremely likely or likely to accept a chairside screening in the future (n = 169). The main reason cited for accepting ORHT was that it was free (n = 138). Conclusion: In order to ensure everyone living with HIV is aware of their infection, HIV testing should be expanded into non-traditional settings. The dental setting may help achieve this important public health milestone.

Contexte: Le test de dépistage rapide du VIH par voie orale (TDRVO) a été mis en place dans le milieu dentaire pour que les gens soient sensibilisés à la possibilité d'une infection au VIH non diagnostiquée. Toutefois, les données du processus de dépistage et les caractéristiques des clients disposés à recevoir le TDRVO restent à être recueillies systématiquement. Description du cas: Le TDRVO a été mis en place dans 3 cliniques d'hygiène dentaire situées dans des établissements scolaires, de mars 2016 à avril 2017. Résultats: 231 personnes ont reçu un TDRVO; elles ont toutes obtenu des résultats non réactifs. La plupart des personnes avaient consulté un prestataire de soins primaires au cours de la dernière année (n = 130), avaient précédemment obtenu un TDRVO (n = 111) et s'étaient décrites comme étant extrêmement susceptibles ou susceptibles à accepter un dépistage au fauteuil à l'avenir (n = 169). La gratuité du TDRVO demeure la raison principale pour avoir accepté le test (n = 138). Conclusion: Le test de dépistage du VIH devrait être offert dans des milieux non traditionnels pour veiller à ce que les gens vivant avec le VIH soient conscients de leur infection. Le milieu dentaire peut aider à franchir cette étape importante de santé publique.

Using Stage- and Slit-Scanning to Improve Contrast and Optical Sectioning in Dual-View Inverted Light Sheet Microscopy (diSPIM).

Dual-view inverted selective plane illumination microscopy (diSPIM) enables high-speed, long-term, four-dimensional (4D) imaging with isotropic spatial resolution. It is also compatible with conventional sample mounting on glass coverslips. However, broadening of the light sheet at distances far from the beam waist and sample-induced scattering degrades diSPIM contrast and optical sectioning. We describe two simple improvements that address both issues and entail no additional hardware modifications to the base diSPIM. First, we demonstrate improved diSPIM sectioning by keeping the light sheet and detection optics stationary, and scanning the sample through the stationary light sheet (rather than scanning the broadening light sheet and detection plane through the stationary sample, as in conventional diSPIM). This stage-scanning approach allows a thinner sheet to be used when imaging laterally extended samples, such as fixed microtubules or motile mitochondria in cell monolayers, and produces finer contrast than does conventional diSPIM. We also used stage-scanning diSPIM to obtain high-quality, 4D nuclear datasets derived from an uncompressed nematode embryo, and performed lineaging analysis to track 97% of cells until twitching. Second, we describe the improvement of contrast in thick, scattering specimens by synchronizing light-sheet synthesis with the rolling, electronic shutter of our scientific complementary metal-oxide-semiconductor (sCMOS) detector. This maneuver forms a virtual confocal slit in the detection path, partially removing out-of-focus light. We demonstrate the applicability of our combined stage- and slit-scanning- methods by imaging pollen grains and nuclear and neuronal structures in live nematode embryos. All acquisition and analysis code is freely available online.

Willingness of Australian health educators and health promotion officers to conduct rapid HIV testing.

Issue addressed: Rapid HIV tests were recently approved by the Australian government. This paper examines the attitudes and willingness to conduct rapid HIV testing (RHT) of Australian health educators and health promotion officers (HE/HPO) from various settings and disciplines.Methods: The aim of the Promoting Research on Methods in Screening Expertise study was to explore knowledge of HIV, attitudes towards people living with HIV, and willingness to conduct RHT among HE/HPO in Australia; this information was attained via an online survey of HE/HPO. Descriptive statistics, χ2 tests, t-tests, and multivariate logistic regression were then conducted.Results: Data from 156 HE/HPOs were analysed. Overall, 60% of participants believed that HE/HPO should offer RHT. Additionally, 70% were personally willing to undergo training in conducting RHT. Fifty-nine percent of participants scored as having 'high' HIV knowledge (at least 12 out of 13 correct answers), with 32% answering all questions correctly. Knowledge was strongly associated with willingness to be trained to conduct RHT.Conclusions: HE/HPO with advanced training in developing evidence-based approaches to improve the health and wellbeing of marginalised and disadvantaged groups may be an appropriate workforce to train to conduct RHT and counselling.So what?: As government agencies and community-based organisations plan to expand RHT, HE/HPO, including those who work outside of sexual health, should be considered as a viable workforce to upskill.

Knowledge of HIV and willingness to conduct oral rapid HIV testing among dentists in Xi'an China.

INTRODUCTION: China is considered a country of low HIV prevalence (780,000 people living with HIV), however, HIV infections among high-risk populations continue to grow at alarming rates. Voluntary Counseling and Testing services were first implemented in 2003, and oral rapid HIV testing (ORHT) began in 2012. Dentists, as oral health experts, would be well placed to conduct ORHT. We assessed willingness of dentists to undertake ORHT in their clinical practice. METHODS: A cross-sectional, paper-based survey of dentists from the Xi'an region of China was conducted from April to June 2013. Dentists were recruited from Shaanxi Stomatological Association using a stratified sampling methodology. A 40-item survey was used to measure knowledge of HIV, attitudes toward people living with HIV and willingness to conduct ORHT. RESULTS: 477 dentists completed the survey with a mean HIV knowledge test score of 13.2/18 (SD 1.9). If made available in the dental setting, 276 (57.9%) preferred to use blood to diagnose HIV, only 190 (39.8%) preferred saliva or both. Four hundred and thirty-five (91.2%) thought that ORHT was needed in dental clinics. Female dentists felt more accepting of ORHT than males (93.8% vs. 87.8%; χ2=5.145; p<0.05). 42.6% of the participants who responded thought that lack of education on ORHT for dentists was the most urgent problem to solve for ORHT, 144 (31.3%) thought that lack of support for ORHT from patients was the most urgent problem. There was statistically significant difference among dental hospital, dentistry and department of dentistry (χ2=24.176; p<0.05). CONCLUSIONS: The majority of Chinese dentists thought that ORHT was needed in the dental setting. Providing opportunities for dentists and dental students to learn about HIV testing guidelines and practices is needed as well as feasibility and implementation science research.

The role of dental hygienists in conducting rapid HIV testing.

PURPOSE: In the U.S., an estimated 21% of people living with HIV/AIDS do not know their positive HIV status. Expanding rap id HIV testing in the dental setting may increase the number of individuals who are aware of their HIV status and can begin medical care and social support services if seropositive and appropriate. As a member of the dental team, the dental hygienist, with the proper knowledge and training, may be suitable to conduct rapid HIV testing.