CD40 Expression by B cells is Required for Optimal Immunity to Murine Pneumocystis Infection.

CD40-CD40L interactions are critical for controlling Pneumocystis infection. However, which CD40-expressing cell populations are important for this interaction have not been well-defined. We used a cohousing mouse model of Pneumocystis infection, combined with flow cytometry and qPCR, to examine the ability of different populations of cells from C57BL/6 mice to reconstitute immunity in CD40 knockout (KO) mice. Unfractionated splenocytes, as well as purified B cells, were able to control Pneumocystis infection, while B cell depleted splenocytes and unstimulated bone-marrow derived dendritic cells (BMDCs) were unable to control infection in CD40 KO mice. Pneumocystis antigen-pulsed BMDCs showed early, but limited, control of infection. Consistent with recent studies that have suggested a role for antigen presentation by B cells, using cells from immunized animals, B cells were able to present Pneumocystis antigens to induce proliferation of T cells. Thus, CD40 expression by B cells appears necessary for robust immunity to Pneumocystis.

A narrative inquiry into non-Indigenous medical educators and leaders participation in reconcilatory work.

INTRODUCTION: Globally, medical schools are operationalising policies and programming to address Indigenous health inequities. Although progress has been made, challenges persist. In Canada, where this research is conducted, Indigenous representation within medical schools remains low, leaving a small number of Indigenous advocates leading unprecedented levels of equity-related work, often with insufficient resources. The change needed within medical education cannot fall solely on the shoulders of Indigenous Peoples; non-Indigenous Peoples must also be involved. This work aims to better understand the pathways of those engaged in this work, with careful consideration given to the facilitators and barriers to ongoing engagement. METHODS: Data collection and analysis were informed by narrative inquiry, a methodology that relies on storytelling to uncover nuance and prompt reflection. In this paper, we focus on interview data collected from Canadian non-Indigenous medical educators and leaders (n = 10). Participants represented different career stages, (early to late career) and occupied a mix of clinical, administrative and education roles. RESULTS: Although each participant's entry into reconciliatory work was unique, we identified common drivers actuating their engagement. Oftentimes their participation was tied to administrative work or propelled by experiences within their roles that forced them to confront the systemic inequalities borne by Indigenous Peoples in both academic and healthcare settings. Some admitted to struggling with understanding their appropriate role in Indigenous reconciliation; their participation often proceeded without firm support. CONCLUSION: Medical schools have an obligation to ensure their faculty, including non-Indigenous Peoples, are equipped to fulfil social accountability mandates regarding Indigenous health. Our findings generate a better understanding of the tensions inherent in this equity work. We urge others to reflect on their role in Indigenous reconciliation, or else medical schools risk generating a false sense of individual and institutional progress.

Emerging adult perceptions of higher-risk cannabis consumption behaviours.

BACKGROUND: Emerging adults have the highest cannabis consumption rates in Canada and are among the most vulnerable to cannabis-related harms. Since certain cannabis consumption behaviours carry greater risks of harm, the Lower-Risk Cannabis Use Guidelines (LRCUG) provide harm reduction strategies. To address a critical gap in the literature, the current study examined emerging adults' awareness of the guidelines and perceptions of higher-risk cannabis consumption behaviours identified within the LRCUG. METHODS: Emerging adults (N = 653) between the ages of 18-25 years were recruited from across Canada. Participants were presented with five vignettes depicting a character's cannabis consumption behaviours. Each vignette focused on a unique aspect of the character's consumption (frequency, polysubstance use, family history of mental illness, method of consumption, and potency). Participants were randomly assigned to one of three conditions within each of the five vignettes that were altered to capture varying levels of risk (e.g. weekly, almost daily, or daily consumption). Following each vignette, participants were asked to respond to four items relating to overall risk of harm, cognitive health, physical health, and mental health. RESULTS: Participants perceived: (1) frequent consumption to be associated with greater risks than less frequent consumption; (2) simultaneous consumption of cannabis and tobacco as being associated with higher risk of harm, yet no difference between simultaneous consumption of cannabis and alcohol or cannabis consumption alone; (3) consuming cannabis with a family history of psychosis or substance use disorder as being associated with greater overall risk than consumption with no family history; (4) smoking and vaping cannabis as associated with more risk than ingesting edibles; and (5) higher-potency THC-dominant strains as being associated with more risk than lower-potency CBD-dominant strains, yet no difference between the two higher-potency THC-dominant strains. CONCLUSIONS: While emerging adults seemed to appreciate the risks associated with some cannabis consumption behaviours, they had difficulty identifying appropriate levels of harm of other higher-risk behaviours. Through an improved understanding of emerging adult perceptions, effective education campaigns should be designed to improve the awareness of cannabis risks and encourage the uptake of harm reduction awareness and strategies.

Canadian cannabis education resources to support youth health literacy: A scoping review and environmental scan.

Background: The 2018 legalisation of cannabis in Canada sparked concern and conversation about the potential negative impacts of youth cannabis use. It is clear that young people are already engaging in cannabis use for a variety of reasons; therefore, youth cannabis education is desirable to promote harm reduction and reduce the risk of adverse physical and mental health outcomes. Objective: To identify and categorise Canadian cannabis education resources using a social-ecological approach informed by the youth health literacy framework, considering multiple factors at the micro-, meso- and macro-levels that influence health literacy and impact behaviour. Methods: In line with scoping review methodology, database searches and an environmental scan of materials were completed. Specific inclusion criteria were identified to encompass all Canadian cannabis education resources directed towards young people aged 9-18 years and adults in contact with youth. Results: A total of 60 resources were identified and categorised using the youth health literacy framework in terms of their focus on (1) micro influences (resources for youth); (2) meso influences (resources for teachers, parents, mentors); and (3) macro influences (resources for indigenous communities and medical professionals). Conclusions: While many resources were identified, issues exist with the accessibility, quality and multicultural considerations of such resources, warranting the development of comprehensive, evidence-based and harm reduction-focused cannabis education for youth.

Clinical Integration of a Highly Accurate Polymerase Chain Reaction Point-of-Care Test Can Inform Immediate Treatment Decisions for Chlamydia, Gonorrhea, and Trichomonas.

BACKGROUND: Accurate same-day sexually transmitted infection (STI) diagnostic testing is generally unavailable, leading to syndromic management with high rates of overtreatment and undertreatment. We analyzed the ease of integration of the Visby STI Panel into clinical practice, studied acceptance by patients and clinic personnel, and assessed the potential to inform accurate treatment decisions. METHODS: In a cross-sectional single-visit study of 55 women aged 18 to 56 years, women self-collected vaginal swab samples that were analyzed using the Visby STI Panel for Chlamydia trachomatis, Neisseria gonorrhoeae (NG), and Trichomonas vaginalis (TV). Results were compared with standard-of-care clinic results from send-out laboratory polymerase chain reaction tests. Surveys assessed patient and device operator experiences with the Visby STI Panel and clinicians' perceived need for and acceptance of the device. Time parameters were measured to evaluate the impact on clinical workflow, and syndromic treatment decisions were compared with anticipated treatment based on the Visby STI Panel results. RESULTS: Patients strongly agreed that sample self-collection was easy, and operators reported the device easy to use. Clinicians valued the rapid return of results, and patients were comfortable waiting up to 30 minutes to receive them. In 13 of 15 cases, the Visby STI Panel correctly identified undertreated patients as infected and correctly identified all 33 incidences of overtreatment. CONCLUSIONS: Clinical adoption of the Visby STI Panel into primary care clinics and doctors' offices could reduce overtreatment and undertreatment of STIs. If integrated efficiently into the clinical workflow, the test would have minimal impact on staff time and visit duration for patients.

Drivers of purchase decisions for cannabis products among consumers in a legalized market: a qualitative study.

BACKGROUND: Cannabis was legalized in Canada for non-medical use in 2018. The goal of legalization was to improve health and safety by creating access to regulated products, with accurate product labels and warnings and no risk of contamination. However, more than 2 years post-legalization, a large proportion of purchases are still suspected to be through unlicensed retailers. This study sought to identify the factors that influenced the purchase decisions of cannabis consumers in Newfoundland and Labrador (NL). METHODS: Semi-structured focus groups and interviews were conducted in NL with individuals who were > 19 and had purchased cannabis within the last 12 months. All sessions were conducted virtually, audio-recorded, and transcribed. A thematic analysis was conducted, and two members of the research team coded the data using NVivo. A combination of deductive and inductive coding was carried out, themes from the literature were identified, and new themes from the transcripts were discovered. A final coding template of the data was agreed upon by the team through discussion and consensus. RESULTS: A total of 23 individuals (30% women) participated, with 83% coming from urban areas. While all cannabis product types were discussed, the conversation naturally focused on dried flower products. Participants discussed a variety of considerations when making purchase decisions categorized around five broad themes: 1) price, 2) quality, 3) packaging and warnings, 4) the source of the cannabis, and 5) social influences. The price difference between licensed and un-licensed sources was commonly discussed as a factor that influenced purchase decisions. Product quality characteristics (e.g. size, color, moisture content) and social influences were also considered in purchase decisions. Participants were generally indifferent to packaging and warning labels but expressed concern about the excessive packaging required for regulated products. CONCLUSION: This study explores the many attributes that influence purchase decisions for dried leaf cannabis. Understanding the drivers of purchase decisions can help inform policy reforms to make regulated cannabis products more appealing to consumers. Further research is needed to measure the effect of each attribute on cannabis purchase decisions.

Driving under the influence of cannabis: perceptions from Canadian youth.

BACKGROUND: Cannabis legalization is intended to protect the public from potential harm by restricting access and promoting greater awareness of cannabis-related risks. Youth are at a greater risk for experiencing road-related harms due to their own or others' use of cannabis. This qualitative research explored youths' perceptions about cannabis and road safety. METHODS: A qualitative study using focus groups (FG) was conducted with youth (age 13-18) and young adults (age 19-25) who resided in Newfoundland and Labrador. Using semi-structured interview questions, the facilitator asked participants to share their opinions about cannabis and road safety. All sessions were hosted virtually using Zoom with recruitment until saturation was met. All sessions were audio recorded, de-identified, and transcribed. Analysis utilized an inductive thematic approach informed by Braun and Clarke's (2006) method and inductive coding was facilitated using NVivo. RESULTS: Six youth (n = 38) and five young adult (n = 53) FG were conducted. Five prominent themes emerged throughout discussions across both age groups including: a) normalization of driving under the influence of cannabis, b) knowledge and awareness, c) perceptions of risk, d) modes of transportation, and e) detection. Variation in perceptions appeared to be influenced by lack of awareness of the impact of cannabis on driving ability, residence in urban versus rural locations, type of vehicle driven (e.g., car vs. off-road vehicles), and gender. CONCLUSION: The themes uncovered from this research will help inform future enhancement of cannabis policy to ensure the safety of all citizens. These findings will also support the inclusion of youth-focused education that will equip youth with informed decision-making strategies regarding road safety. Furthermore, these findings can be utilized to inform the refinement of cannabis driving policies to ensure the safety of all citizens on or off the road.

An Update on Perinatal Cannabis Use.

Since the legalization of non-medical cannabis, many questions have arisen regarding cannabis use during pregnancy. Obstetrical care providers can minimize the harms of prenatal cannabis use using evidence-based information. This commentary describes populations at highest risk for use, the predictors of use, the reasons birthing people continue or stop using cannabis during pregnancy, and short- and long-term outcomes for infants and children exposed to cannabis prenatally. Our goal is to equip providers with knowledge from the literature to inform practical decision-making.

MUC1 mediates Pneumocystis murina binding to airway epithelial cells.

Previous studies have shown that Pneumocystis binds to pneumocytes, but the proteins responsible for binding have not been well defined. Mucins are the major glycoproteins present in mucus, which serves as the first line of defence during airway infection. MUC1 is the best characterised membrane-tethered mucin and is expressed on the surface of most airway epithelial cells. Although by electron microscopy Pneumocystis primarily binds to type I pneumocytes, it can also bind to type II pneumocytes. We hypothesized that Pneumocystis organisms can bind to MUC1 expressed by type II pneumocytes. Overexpression of MUC1 in human embryonic kidney HEK293 cells increased Pneumocystis binding, while knockdown of MUC1 expression by siRNA in A549 cells, a human adenocarcinoma-derived alveolar type II epithelial cell line, decreased Pneumocystis binding. Immunofluorescence labelling indicated that MUC1 and Pneumocystis were co-localised in infected mouse lung tissue. Incubation of A549 cells with Pneumocystis led to phosphorylation of ERK1/2 that increased with knockdown of MUC1 expression by siRNA. Pneumocystis caused increased IL-6 and IL-8 secretion by A549 cells, and knockdown of MUC1 further increased their secretion in A549 cells. Taken together, these results suggest that binding of Pneumocystis to MUC1 expressed by airway epithelial cells may facilitate establishment of productive infection.

Pharmacists' perceptions of the Canadian opioid regulatory exemptions on patient care and opioid stewardship.

BACKGROUND: This study explored the perceptions of Canadian pharmacists about the barriers and facilitators of providing opioid stewardship activities in pharmacy practice, considering the subsection 56(1) class exemption under Health Canada's Controlled Drugs and Substances Act (CDSA). METHODS: Qualitative key informant telephone interviews were conducted with a convenience sample of pharmacists from across Canada. We included community or primary health care team-based pharmacists who self-identified as having experience with providing care for patients using opioids via the exemptions. All transcripts were de-identified, and thematic analysis was conducted to identify themes. Ethics approval was obtained. RESULTS: Twenty pharmacists from community and primary health care teams, from all provinces and from urban and rural practices were interviewed. The following themes emerged: 1) optimization of opioid-related patient care, 2) jurisdictional impact and 3) awareness and education. Barriers and facilitators for opioid stewardship activities were identified. DISCUSSION: The exemptions facilitated the pharmacists' ability to provide opioid stewardship and positively affect patient care by providing continuity of and timely access to care. Our research demonstrated that pharmacists can responsibly and independently manage opioid prescriptions within this expanded scope, demonstrating the valuable contribution pharmacists can have in opioid stewardship. CONCLUSION: Pharmacists were willing and able to care for patients receiving opioid medication and thereby played a role in helping address the opioid crisis. The benefits of these exemptions were demonstrated beyond situations related to the COVID-19 pandemic and warrant consideration for consistent implementation across provincial and territorial jurisdictions, thereby ensuring equitable access to care for all Canadians.

Too young for Cannabis? Choice of minimum legal age for legalized non-medical Cannabis in Canada.

BACKGROUND: Choice of minimum legal age (MLA) for cannabis use is a critical and contentious issue in legalization of non-medical cannabis. In Canada where non-medical cannabis was recently legalized in October 2018, the federal government recommended age 18, the medical community argued for 21 or even 25, while public consultations led most Canadian provinces to adopt age 19. However, no research has compared later life outcomes of first using cannabis at these different ages to assess their merits as MLAs. METHODS: We used doubly robust regression techniques and data from nationally representative Canadian surveys to compare educational attainment, cigarette smoking, self-reported general and mental health associated with different ages of first cannabis use. RESULTS: We found different MLAs for different outcomes: 21 for educational attainment, 19 for cigarette smoking and mental health and 18 for general health. Assuming equal weight for these individual outcomes, the 'overall' MLA for cannabis use was estimated to be 19 years. Our results were robust to various robustness checks. CONCLUSION: Our study indicated that there is merit in setting 19 years as MLA for non-medical cannabis.

Differences in hospital length of stay between neonates exposed to buprenorphine versus methadone in utero: A retrospective chart review.

INTRODUCTION: Neonatal abstinence syndrome is a growing concern in neonatal intensive care units in rural and remote settings. METHODS: A retrospective chart review was conducted of 180 mother-infant dyads born with in utero exposure to buprenorphine (n=60), methadone (n=60) or to other opioids (n=60) to determine neonatal length of stay in hospital, number of days on morphine, day of life of initiation of morphine and the need for phenobarbital. RESULTS: The length of stay in hospital for neonates was 5.8 days shorter (95% confidence interval [CI] 6.1 to 8.5 days) for buprenorphine exposure in utero compared to methadone (P=0.001). For neonates requiring treatment for Neonatal abstinence syndrome, those with in utero exposure to buprenorphine required 6.1 fewer days (95% CI 2.5 to 9.7) of treatment with morphine then those exposed to methadone (P<0.0005). There were no statistically significant differences in day of life of initiation of morphine therapy for each of the study groups. The proportion of neonates requiring adjuvant therapy with phenobarbital was statistically significantly higher in neonates exposed to methadone in utero than either buprenorphine or illicit opioids (P<0.0005). CONCLUSIONS: Retrospective data suggest that neonates with in utero exposure to buprenorphine experience a shorter length of stay in hospital, fewer days of treatment with morphine for neonatal abstinence syndrome, and less use of phenobarbital than neonates exposed in utero to methadone. This suggests that Ontario provincial guidelines should be updated to recommend buprenorphine as first line for replacement therapy in pregnancy.

Characterization of p57, a Stage-Specific Antigen of Pneumocystis murina.

Pneumocystis has a large multicopy gene family encoding proteins related to the major surface glycoprotein (Msg), whose functions are largely unknown. We expressed one such protein of Pneumocystis murina, p57, which is encoded by 3 highly conserved genes, and demonstrated by immunoblot that immunocompetent mice that were immunized with crude Pneumocystis antigens or that had cleared Pneumocystis infection developed antibodies to p57. Using hyperimmune anti-p57 serum combined with immunolabeling, we found that p57 was expressed by small trophic forms and intracystic bodies, whereas it was not expressed on larger trophic forms or externally by cysts. Expression of p57 and Msg by trophic forms was largely mutually exclusive. Treatment of infected animals with caspofungin inhibited cyst formation and markedly decreased p57 expression. While p57 expression was seen in immunocompetent mice infected with Pneumocystis, immunization with recombinant p57 did not result in altered cytokine expression by lymphocytes or in diminished infection in such mice. Thus, p57 appears to be a stage-specific antigen of Pneumocystis that is expressed on intracystic bodies and young trophic forms and may represent a mechanism to conserve resources in organisms during periods of limited exposure to host immune responses.

The Major Surface Glycoprotein of Pneumocystis murina Does Not Activate Dendritic Cells.

The major surface glycoprotein (Msg) is the most abundant surface protein among Pneumocystis species. Given that Msg is present on both the cyst and trophic forms of Pneumocystis and that dendritic cells play a critical role in initiating host immune responses, we undertook studies to examine activation of bone marrow-derived myeloid dendritic cells by Msg purified from Pneumocystis murina. Incubation of dendritic cells with Msg did not lead to increased expression of CD40, CD80, CD86, or major histocompatibility complex class II or to increased secretion of any of 10 cytokines. Microarray analysis identified very few differentially expressed genes. In contrast, lipopolysaccharide-activated dendritic cells had positive results of all of these assays. However, Msg did bind to mouse mannose macrophage receptor and human DC-SIGN, 2 C-type lectins expressed by dendritic cells that are important in recognition of pathogen-associated high-mannose glycoproteins. Deglycosylation of Msg demonstrated that this binding was dependent on glycosylation. These studies suggest that Pneumocystis has developed a mechanism to avoid activation of dendritic cells, potentially by the previously identified loss of genes that are responsible for the high level of protein mannosylation found in other fungi.

Caribbean Wellness Day: promoting a region-wide day of action.

In response to the epidemic of non-communicable diseases (NCDs) in the Caribbean, the Heads of Government of the Caribbean Community (CARICOM) issued the 2007 Port-of-Spain Declaration, "Uniting to Stop the Epidemic of Chronic Non-communicable Diseases" and declared the second Saturday in September to be "Caribbean Wellness Day" (CWD). CWD is a call-to-action for engaging the population of the Member States in confronting NCDs and their risk factors. This report reviews the genesis, implementation, and institutionalization of CWD in CARICOM and beyond. The study used quantitative and qualitative methods, including 29 surveys, 7 in-depth interviews, and analysis of media content. Data was provided by NCD focal points at the Ministry of Health in 15 CARICOM countries, as well as by 7 non-governmental and 4 private organizations. Branding materials were well received and locally adapted; much of CWD media content originated from organization websites and were not community-based. Events typically focused on physical activity and health screenings and were attended by up to 3 000 participants. Though most were held in city centers, some CWD activities have involved rural and indigenous populations. CWD has become a catalyst for multisectoral engagement and health promotion activities. Inspired by CWD, PAHO initiated "Wellness Week in the Americas," which includes CWD and promotes its tenets across the Region of the Americas. As CWD further develops, consideration should be given to reliable, adequate, and sustainable financing; to measuring and evaluating its impact on NCDs; and to widening its reach to include those outside of city centers.

En respuesta a la epidemia de las enfermedades no transmisibles en el Caribe, los jefes de Gobierno de la Comunidad del Caribe (CARICOM) emitieron en el 2007 la Declaración de Puerto España, "Unidos para detener la epidemia de las enfermedades crónicas no transmisibles", y propusieron instaurar el segundo sábado de septiembre como el "Día del Bienestar en el Caribe". Este día es un llamado a la acción para fomentar la participación la población de los Estados Miembros contra las enfermedades no transmisibles y sus factores de riesgo.Este informe examina la génesis, la puesta en marcha y la institucionalización del Día del Bienestar en el Caribe en CARICOM y otros lugares. En el estudio se emplearon métodos cuantitativos y cualitativos, entre ellos 29 encuestas, 7 entrevistas exhaustivas y el análisis del contenido de medios de difusión. Los datos fueron proporcionados por puntos focales para las enfermedades no transmisibles del Ministerio de Salud de 15 países de CARICOM, así como por siete organizaciones no gubernamentales y cuatro organizaciones privadas.Los materiales de imagen institucional fueron bien recibidos y se adaptaron localmente; gran parte del contenido sobre el Día del Bienestar en los medios de difusión provenía de los sitios web de organizaciones y no de las comunidades. Los eventos se centraron generalmente en la actividad física y los exámenes médicos de tamizaje, y asistieron hasta 3 000 personas. Aunque la mayoría se celebraron en el centro de las ciudades, algunas actividades se han realizado con poblaciones rurales e indígenas.El Día del Bienestar en el Caribe se ha convertido en un catalizador para la participación multisectorial y las actividades de promoción de la salud. Inspirada en él, la OPS instituyó la "Semana del Bienestar en las Américas", que incluye a su vez el Día del Bienestar en el Caribe y difunde sus principios en toda la Región de las Américas. Conforme el Día del Bienestar en el Caribe evolucione aún más, debe pensarse en obtener un financiamiento confiable, adecuado y sostenible; en medir y evaluar sus repercusiones en materia de enfermedades no transmisibles; y en ampliar su alcance para abarcar a quienes viven lejos del centro de las ciudades.

Em resposta à epidemia de doenças não transmissíveis (DNTs) no Caribe, os chefes de governo da Comunidade do Caribe (CARICOM) emitiram a Declaração de Porto de Espanha de 2007, "Unidos para deter a epidemia de doenças crônicas não transmissíveis", e declararam o segundo sábado de setembro o "Dia do Bem-estar no Caribe". Trata-se de uma chamada à ação aos Estados Membros para engajar a população a enfrentar as DNTs e os fatores de risco associados.Este informe faz uma análise da origem, implantação e institucionalização do Dia do Bem-Estar no Caribe no CARICOM e outras regiões. Neste estudo foram empregados métodos quantitativos e qualitativos, com 29 inquéritos, 7 entrevistas aprofundadas e uma análise de conteúdo de mídias. Os dados foram fornecidos pelos centros de coordenação de DNTs nos ministérios da Saúde de 15 países da CARICOM, assim como por sete organizações não governamentais e quatro entidades privadas.Os materiais de branding foram bem aceitos e adaptados ao contexto local. Grande parte de conteúdo de mídia do Dia do Bem-Estar no Caribe foi proveniente de sites de entidades e não foram originados nas comunidades. Os eventos normalmente envolveram atividade física e exames de prevenção de saúde, com o comparecimento de até 3.000 participantes. Apesar de a maioria dos eventos ter sido realizada em centros urbanos, ocorreram algumas atividades para populações rurais e indígenas.O Dia do Bem-Estar no Caribe é um incentivador para a participação multissetorial e para atividades de promoção da saúde. Inspirada nesta iniciativa, a Organização Pan-Americana da Saúde (OPAS) instituiu a "Semana do Bem-Estar nas Américas", englobando e promovendo os princípios do Dia do Bem-Estar no Caribe em toda a Região das Américas. Com a expansão deste acontecimento, é preciso considerar como custeá-lo de maneira adequada, segura e sustentável, mensurar e avaliar a repercussão que ele tem e ampliar o seu alcance para chegar às populações fora dos centros urbanos.

Evaluating policy responses to noncommunicable diseases in seven Caribbean countries: challenges to addressing unhealthy diets and physical inactivity.

OBJECTIVE: To identify, assess, and compare existing policies on noncommunicable diseases (NCDs) in the Caribbean, gaps in policy responses, and the factors influencing successful policy development and implementation following the Port of Spain Declaration of 2007. Specifically, to examine policies that target the upstream determinants of two NCD risk factors-unhealthy diets and physical inactivity. METHODS: A total of 76 semi-structured interviews with 80 relevant stakeholders in government, the private sector, and civil society were complemented by policy document analysis. Interviews were analyzed pragmatically, framed by the CARICOM government commitments, the WHO NCD Action Plan, a Multiple Streams framework approach, and realist evaluation ideas. RESULTS: The most widely-reported policy successes involved health promotion activities (e.g., school meal programs) that leveraged multisectoral collaboration among government ministries, such as Health, Education, and Agriculture. Large policy gaps still exist around creating legislative, physical, and social environments to support healthy eating and physical activity at the population level. Multisectoral NCD commissions successfully reached across sectors, but had limited influence on policy development. Different policy levels emerged with national-level policies considered a lengthy process, while "On-the-ground" programming was considered faster to implement than national policies. External barriers included a reliance on food imports enabled by international trade agreements limited availability, quality, and affordability of healthy foods. International pushback limited legislation to reduce food imports and the absence of an international/regional framework, similar to the Framework Convention on Tobacco Control, further impedes efforts. CONCLUSIONS: Regional collaboration and political support across sectors are essential to accelerating the pace of action to support healthy eating and active living environments. Policy "blueprints" could accelerate the process of development. Regional "NCD champions" could spearhead such responses and approaches.

Pulmonary Interleukin-17-Positive Lymphocytes Increase during Pneumocystis murina Infection but Are Not Required for Clearance of Pneumocystis.

Pneumocystis remains an important pathogen of immunosuppressed patients, causing a potentially life-threatening pneumonia. Despite its medical importance, the immune responses required to control infection, including the role of interleukin-17 (IL-17), which is important in controlling other fungal infections, have not been clearly defined. Using flow cytometry and intracellular cytokine staining after stimulation with phorbol myristate acetate and ionomycin, we examined gamma interferon (IFN-γ), IL-4, IL-5, and IL-17 production by lung lymphocytes in immunocompetent C57BL/6 mice over time following infection with Pneumocystismurina We also examined the clearance of Pneumocystis infection in IL-17A-deficient mice. The production of both IFN-γ and IL-17 by pulmonary lymphocytes increased during infection, with maximum production at approximately days 35 to 40, coinciding with peak Pneumocystis levels in the lungs, while minimal changes were seen in IL-4- and IL-5-positive cells. The proportion of cells producing IFN-γ was consistently higher than for cells producing IL-17, with peak levels of ∼25 to 30% of CD3+ T cells for the former compared to ∼15% for the latter. Both CD4+ T cells and γδ T cells produced IL-17. Administration of anti-IFN-γ antibody led to a decrease in IFN-γ-positive cells, and an increase in IL-5-positive cells, but did not impact clearance of Pneumocystis infection. Despite the increases in IL-17 production during infection, IL-17A-deficient mice cleared Pneumocystis infection with kinetics similar to C57BL/6 mice. Thus, while IL-17 production in the lungs is increased during Pneumocystis infection in immunocompetent mice, IL-17A is not required for control of Pneumocystis infection.

Conflicting Selection Pressures Will Constrain Viral Escape from Interfering Particles: Principles for Designing Resistance-Proof Antivirals.

The rapid evolution of RNA-encoded viruses such as HIV presents a major barrier to infectious disease control using conventional pharmaceuticals and vaccines. Previously, it was proposed that defective interfering particles could be developed to indefinitely control the HIV/AIDS pandemic; in individual patients, these engineered molecular parasites were further predicted to be refractory to HIV's mutational escape (i.e., be 'resistance-proof'). However, an outstanding question has been whether these engineered interfering particles-termed Therapeutic Interfering Particles (TIPs)-would remain resistance-proof at the population-scale, where TIP-resistant HIV mutants may transmit more efficiently by reaching higher viral loads in the TIP-treated subpopulation. Here, we develop a multi-scale model to test whether TIPs will maintain indefinite control of HIV at the population-scale, as HIV ('unilaterally') evolves toward TIP resistance by limiting the production of viral proteins available for TIPs to parasitize. Model results capture the existence of two intrinsic evolutionary tradeoffs that collectively prevent the spread of TIP-resistant HIV mutants in a population. First, despite their increased transmission rates in TIP-treated sub-populations, unilateral TIP-resistant mutants are shown to have reduced transmission rates in TIP-untreated sub-populations. Second, these TIP-resistant mutants are shown to have reduced growth rates (i.e., replicative fitness) in both TIP-treated and TIP-untreated individuals. As a result of these tradeoffs, the model finds that TIP-susceptible HIV strains continually outcompete TIP-resistant HIV mutants at both patient and population scales when TIPs are engineered to express >3-fold more genomic RNA than HIV expresses. Thus, the results provide design constraints for engineering population-scale therapies that may be refractory to the acquisition of antiviral resistance.

Engaging patients in health research: identifying research priorities through community town halls.

BACKGROUND: The vision of Canada's Strategy for Patient-Oriented Research is that patients be actively engaged as partners in health research. Support units have been created across Canada to build capacity in patient-oriented research and facilitate its conduct. This study aimed to explore patients' health research priorities in the province of Newfoundland and Labrador (NL). METHODS: Eight town halls were held with members of the general public in rural and urban settings across the province. Sessions were a hybrid information-consultation event, with key questions about health research priorities and outcomes guiding the discussion. RESULTS: Sixty eight members of the public attended town hall sessions. A broad range of health experiences in the healthcare system were recounted. Key priorities for the public included access and availability of providers and services, disease prevention and health promotion, and follow-up support and community care. In discussing their health research priorities, participants spontaneously raised a broad range of suggestions for improving the healthcare system in our jurisdiction. CONCLUSIONS: Public research priorities and suggestions for improving the provision of healthcare provide valuable information to guide Support Units' planning and priority-setting processes. A range of research areas were raised as priorities for patients that are likely comparable to other healthcare systems. These create a number of health research questions that would be in line with public priorities. Findings also provide lessons learned for others and add to the evidence base on patient engagement methods.

ß-Glucans Are Masked but Contribute to Pulmonary Inflammation During Pneumocystis Pneumonia.

ß-glucans, which can activate innate immune responses, are a major component in the cell wall of the cyst form of Pneumocystis In the current study, we examined whether ß-1,3-glucans are masked by surface proteins in Pneumocystis and what role ß-glucans play in Pneumocystis-associated inflammation. For 3 species, including Pneumocystis jirovecii, which causes Pneumocystis pneumonia in humans, Pneumocystis carinii, and Pneumocystis murina, ß-1,3-glucans were masked in most organisms, as demonstrated by increased exposure following trypsin treatment. Using quantitative polymerase chain reaction and microarray techniques, we demonstrated in a mouse model of Pneumocystis pneumonia that treatment with caspofungin, an inhibitor of ß-1,3-glucan synthesis, for 21 days decreased expression of a broad panel of inflammatory markers, including interferon γ, tumor necrosis factor α, interleukin 1ß, interleukin 6, and multiple chemokines/chemokine ligands. Thus, ß-glucans in Pneumocystis cysts are largely masked, which likely decreases innate immune activation; this mechanism presumably was developed for interactions with immunocompetent hosts, in whom organism loads are substantially lower. In immunosuppressed hosts with a high organism burden, organism death and release of glucans appears to be an important contributor to deleterious host inflammatory responses.