A localizing nanocarrier formulation enables multi-target immune responses to multivalent replicating RNA with limited systemic inflammation.

RNA vaccines possess significant clinical promise in counteracting human diseases caused by infectious or cancerous threats. Self-amplifying replicon RNA (repRNA) has been thought to offer the potential for enhanced potency and dose sparing. However, repRNA is a potent trigger of innate immune responses in vivo, which can cause reduced transgene expression and dose-limiting reactogenicity, as highlighted by recent clinical trials. Here, we report that multivalent repRNA vaccination, necessitating higher doses of total RNA, could be safely achieved in mice by delivering multiple repRNAs with a localizing cationic nanocarrier formulation (LION). Intramuscular delivery of multivalent repRNA by LION resulted in localized biodistribution accompanied by significantly upregulated local innate immune responses and the induction of antigen-specific adaptive immune responses in the absence of systemic inflammatory responses. In contrast, repRNA delivered by lipid nanoparticles (LNPs) showed generalized biodistribution, a systemic inflammatory state, an increased body weight loss, and failed to induce neutralizing antibody responses in a multivalent composition. These findings suggest that in vivo delivery of repRNA by LION is a platform technology for safe and effective multivalent vaccination through mechanisms distinct from LNP-formulated repRNA vaccines.

Biochemical and clinical studies of putative allergens to assess what distinguishes them from other non-allergenic proteins in the same family.

Many protein families have numerous members listed in databases as allergens; however, some allergen database entries, herein called "orphan allergens", are members of large families of which all other members are not allergens. These orphan allergens provide an opportunity to assess whether specific structural features render a protein allergenic. Three orphan allergens [Cladosporium herbarum aldehyde dehydrogenase (ChALDH), Alternaria alternata ALDH (AaALDH), and C. herbarum mannitol dehydrogenase (ChMDH)] were recombinantly produced and purified for structure characterization and for clinical skin prick testing (SPT) in mold allergic participants. Examination of the X-ray crystal structures of ChALDH and ChMDH and a homology structure model of AaALDH did not identify any discernable epitopes that distinguish these putative orphan allergens from their non-allergenic protein relatives. SPT results were aligned with ChMDH being an allergen, 53% of the participants were SPT (+). AaALDH did not elicit SPT reactivity above control proteins not in allergen databases (i.e., Psedomonas syringae indole-3-acetaldehyde dehydrogenase and Zea mays ALDH). Although published results showed consequential human IgE reactivity with ChALDH, no SPT reactivity was observed in this study. With only one of these three orphan allergens, ChMDH, eliciting SPT(+) reactions consistent with the protein being included in allergen databases, this underscores the complicated nature of how bioinformatics is used to assess the potential allergenicity of food proteins that could be newly added to human diets and, when needed, the subsequent clinical testing of that bioinformatic assessment.Trial registration number and date of registration AAC-2017-0467, approved as WIRB protocol #20172536 on 07DEC2017 by WIRB-Copernicus (OHRP/FDA Registration #: IRB00000533, organization #: IORG0000432).

High prevalence of Helicobacter pylori clarithromycin resistance mutations among Seattle patients measured by droplet digital PCR.

BACKGROUND: Treatment of Helicobacter pylori infection is often empiric; however, current guidelines for management of Helicobacter pylori infection advise against the use of standard triple therapy (clarithromycin, amoxicillin, and proton-pump inhibitor) when clarithromycin resistance exceeds 20%. We developed and tested a new culture-free assay to detect clarithromycin resistance-conferring mutations to determine the prevalence of H. pylori clarithromycin resistance in patients from the United States Pacific Northwest. MATERIALS AND METHODS: Droplet digital PCR (ddPCR) was used to detect the H. pylori 23S rRNA gene, and resistance-conferring mutations, in archived, formalin-fixed, paraffin-embedded (FFPE) gastric tissue and to retrospectively determine the prevalence of clarithromycin-resistant H. pylori among 110 patients at an academic medical center in the Northwest United States between 2012 and 2014. RESULTS: Of 102 patients with the H. pylori 23S rRNA gene detected by the ddPCR assay, 45 (44%) had clarithromycin resistance mutations. Thirty-three of the 45 patients with clarithromycin resistance mutations had a mix of wild-type and resistance alleles. Prevalence of clarithromycin resistance mutations differed among racial groups and was highest among Asians, with mutations detected in 14 (67%) of the 21 patient samples. CONCLUSIONS: The prevalence of clarithromycin resistance detected in this region exceeds 20%, indicating that standard triple therapy should not be the first-line antibiotic treatment for H. pylori infection. Culture-free assays for detecting clarithromycin resistance mutations can be performed on archived tissue samples and will aid in informing tailored treatment for effective H. pylori eradication.

Access to and Use of the Internet by Veterans with Serious Mental Illness.

This study evaluated internet use among 239 veterans with serious mental illness who completed questionnaires assessing demographics and internet use in 2010-2011. The majority of individuals (70 %) reported having accessed the internet and among those, 79 % had accessed it within the previous 30 days. Those who were younger and more educated were more likely to have accessed the internet, as were those with a schizophrenia spectrum disorder, bipolar disorder, or major depressive disorder, compared to individuals with PTSD. Veterans with serious mental illness commonly use the internet, including to obtain health information, though use varies across demographic characteristics and clinical diagnosis.

Caspase 9 is constitutively activated in mouse oocytes and plays a key role in oocyte elimination during meiotic prophase progression.

In many mammalian species, more than half of the initial oocyte population is eliminated by neonatal life, thus limiting the oocyte reserve for reproduction. The cause or mechanism of this major oocyte loss remains poorly understood. We examined the apoptotic pathway involved in oocyte elimination in wild-type mouse ovaries as well as in Msh5 -/- ovaries, in which all oocytes were eliminated due to a lack of double strand break repair. Immunoblot and immunofluorescence staining showed that an initiator caspase 9 and an effector caspase 7 were constitutively activated in almost all oocytes in fetal ovaries regardless of their genotypes. In caspase 9 -/- ovaries, the total number of oocytes remained high while that in wild-type ovaries steadily declined during ovarian development. Therefore, the activation of caspase 9 was required for but did not immediately lead to oocyte demise. We found that XIAP, an endogenous inhibitor of apoptosis, was also abundant in oocytes during meiotic prophase progression. On the other hand, a cleaved form of PARP1, a target of effector caspases, was localized to the nuclei of a limited number of oocytes, and the frequency of cleaved PARP1-positive oocyte nuclei increased significantly higher before all oocytes disappeared in Msh5 -/- ovaries. We conclude that the mitochondrial apoptotic pathway mediated by caspase 9 is constitutively activated in oocytes and renders the elimination of oocytes with meiotic errors, which can be captured by the cleavage of PARP1.

Understanding the perspective of community co-educators on community-based service learning: a qualitative analysis.

Background: Community-based service learning (CBSL) is a core component of the Canadian medical education system. However, the unique role of community partner organizations (CPOs) in supporting CBSL remains unclear. This qualitative study evaluates the perspective of CPOs as co-educators in the undergraduate medical curriculum. Methods: We conducted eight semi-structured, one-on-one interviews with CPOs at a medical school in Toronto, Ontario between 2020-2021. Interviews were conducted following a pre-determined interview guide and then recorded, de-identified, and transcribed. Three reviewers independently performed an inductive thematic analysis of codes followed by a group review of discrepancies. Results: Five main findings were identified: 1) CPOs share a common interest in serving as co-educators; 2) considerable heterogeneity in the understanding of co-education exists; 3) there is an opportunity for increased partnerships between CPOs and faculty; 4) the role of co-educators is limited by curriculum structure; and 5) co-educators facilitate unique teachings of social determinants of health otherwise not available through traditional didactic teaching. Conclusions: There is an emerging, unique role for community co-educators in the undergraduate medical curriculum, supported by interest from CPOs. Its emphasis may contribute to future cohorts of medical students capable of understanding and addressing the needs of the populations they serve.

Contexte: L'apprentissage par le service communautaire (APSC) est une composante essentielle du système d'éducation médicale canadien. Cependant, le rôle unique des organismes communautaires partenaires (OCP) dans le soutien de l'APSC n'est toujours pas clair. Cette étude qualitative évalue le point de vue des OCP en tant que co-éducateurs dans le programme d'études médicales de premier cycle. Méthodes: Nous avons mené huit entrevues individuelles semi-structurées avec des OCP d'une faculté de médecine de Toronto, en Ontario, entre 2020 et 2021. Les entrevues ont été menées en suivant un guide d'entrevue prédéterminé, puis enregistrées, dépersonnalisées et transcrites. Trois examinateurs ont effectué indépendamment une analyse thématique inductive des codes, suivie d'un examen collectif des divergences. Résultats: Cinq conclusions principales ont été identifiées : 1) les OCP ont un intérêt commun à agir en tant que co-éducateurs; 2) il existe une grande hétérogénéité dans la compréhension de la co-éducation; 3) il est possible d'accroître les partenariats entre les OCP et le corps professoral; 4) le rôle des co-éducateurs est limité par la structure du curriculum médical et 5) les co-éducateurs facilitent des opportunités d'apprentissage uniques sur les déterminants sociaux de la santé qui ne sont pas disponibles dans le cadre de l'enseignement didactique traditionnel. Conclusions: Les co-éducateurs communautaires jouent un rôle novateur et unique dans le programme d'études médicales de premier cycle, soutenu par l'intérêt des OCP. L'accent mis sur ce rôle peut contribuer à former de futures cohortes d'étudiants en médecine capables de comprendre et de répondre aux besoins des populations qu'ils servent.

A self-amplifying RNA vaccine prevents enterovirus D68 infection and disease in preclinical models.

The recent emergence and rapid response to severe acute respiratory syndrome coronavirus 2 was enabled by prototype pathogen and vaccine platform approaches, driven by the preemptive application of RNA vaccine technology to the related Middle East respiratory syndrome coronavirus. Recently, the National Institutes of Allergy and Infectious Diseases identified nine virus families of concern, eight enveloped virus families and one nonenveloped virus family, for which vaccine generation is a priority. Although RNA vaccines have been described for a variety of enveloped viruses, a roadmap for their use against nonenveloped viruses is lacking. Enterovirus D68 was recently designated a prototype pathogen within the family Picornaviridae of nonenveloped viruses because of its rapid evolution and respiratory route of transmission, coupled with a lack of diverse anti-enterovirus vaccine approaches in development. Here, we describe a proof-of-concept approach using a clinical stage RNA vaccine platform that induced robust enterovirus D68-neutralizing antibody responses in mice and nonhuman primates and prevented upper and lower respiratory tract infections and neurological disease in mice. In addition, we used our platform to rapidly characterize the antigenic diversity within the six genotypes of enterovirus D68, providing the necessary data to inform multivalent vaccine compositions that can elicit optimal breadth of neutralizing responses. These results demonstrate that RNA vaccines can be used as tools in our pandemic-preparedness toolbox for nonenveloped viruses.

Search strategies in systematic reviews in periodontology and implant dentistry.

OBJECTIVE: To perform an overview of literature search strategies in systematic reviews (SRs) published in periodontology and implant dentistry. MATERIALS AND METHODS: Two electronic databases (PubMed and Cochrane Database of SRs) were searched, independently and in duplicate, for SRs with meta-analyses on interventions, with the last search performed on 11 November 2012. Manual searches of the reference lists of included SRs and 10 specialty dental journals were conducted. Methodological issues of the search strategies of included SRs were assessed with Cochrane collaboration guidelines and AMSTAR recommendations. The search strategies employed in Cochrane and paper-based SRs were compared. RESULTS: A total of 146 SRs with meta-analyses were included, including 19 Cochrane and 127 paper-based SRs. Some issues, such as "the use of keywords," were reported in most of the SRs (86%). Other issues, such as "search of grey literature" and "language restriction," were not fully reported (34% and 50% respectively). The quality of search strategy reporting in Cochrane SRs was better than that of paper-based SRs for seven of the eight criteria assessed. CONCLUSION: There is room for improving the quality of reporting of search strategies in SRs in periodontology and implant dentistry, particularly in SRs published in paper-based journals.

The Motivation and Pleasure Scale-Self-Report (MAP-SR): reliability and validity of a self-report measure of negative symptoms.

The Clinical Assessment Interview for Negative Symptoms (CAINS) is an empirically developed interview measure of negative symptoms. Building on prior work, this study examined the reliability and validity of a self-report measure based on the CAINS-the Motivation and Pleasure Scale-Self-Report (MAP-SR)-that assesses the motivation and pleasure domain of negative symptoms. Thirty-seven participants with schizophrenia or schizoaffective disorder completed the 18-item MAP-SR, the CAINS, and other measures of functional outcome. Item analyses revealed three items that performed poorly. The revised 15-item MAP-SR demonstrated good internal consistency and convergent validity with the clinician-rated Motivation and Pleasure scale of the CAINS, as well as good discriminant validity, with little association with psychotic symptoms or depression/anxiety. MAP-SR scores were related to social anhedonia, social closeness, and clinician-rated social functioning. The MAP-SR is a promising self-report measure of severity of negative symptoms.

Effect of Peripheral Magnetic Stimulation on Acute and Chronic Pain After Surgery: A Systematic Review and Meta-Analysis.

Peripheral magnetic stimulation (PMS) is a potentially promising modality to help manage postoperative pain. We systematically reviewed the effect of PMS on acute and chronic postoperative pain. MEDLINE, Cochrane CENTRAL, EMBASE, ProQuest Dissertations, and clinical trials.gov were searched from inception until May 2021. We included studies of any study design that included patients ≥18 years of age undergoing any type of surgery that administered PMS within the perioperative period and evaluated postoperative pain. Seventeen randomized controlled trials and 1 nonrandomized clinical trial were included into the review. Thirteen out of the 18 studies found a positive effect with PMS on postoperative pain scores. In our meta-analysis, peripheral magnetic stimulation was more efficacious than sham or no intervention within the first 7 postoperative days (mean difference [MD] -1.64 on a 0 to 10 numerical rating score, 95% confidence interval [CI] -2.08 to -1.20, I2 = 77%, 6 studies, 231 patients). This was also true at 1 and 2 months after surgery (MD -1.82, 95% CI -2.48 to -1.17, I2 = 0%, 3 studies, 104 patients; and MD -1.96, 95% CI -3.67 to -.26, I2 = 84%, 3 studies, 104 patients, respectively). A difference was not seen with persistent pain at 6 and 12-months after surgery, acute postoperative opioid consumption, or adverse events between groups. Results are limited by heterogeneity and generally low-quality studies, as well as low or very low quality of evidence. High-quality and adequately blinded trials are needed to definitively confirm the benefits of peripheral magnetic stimulation administered in the perioperative period. PERSPECTIVE: This review evaluates the efficacy and safety of PMS on postoperative pain. The results help elucidate PMS' role in postoperative pain management and identify gaps where more research is required.

SRY upregulation of SOX9 is inefficient and delayed, allowing ovarian differentiation, in the B6.Y(TIR) gonad.

SRY on the Y-chromosome acts as a transcription factor to initiate testicular differentiation in mammals. Sox9 is a SRY target gene, upregulated immediately after Sry expression, and plays a key role in testicular differentiation. In the present study, we examined the expression of SRY and SOX9 proteins in the B6.Y(TIR) gonad, which undergoes partial or complete sex reversal. The results show that the ontogeny of SRY expression in the B6.Y(TIR) gonad was comparable with that in the B6.XY gonad. On the other hand, while SOX9 expression immediately followed SRY expression in the B6.XY gonad, it was considerably delayed compared to SRY expression in the B6.Y(TIR) gonad or SOX9 expression in the B6.XY gonad. Although SOX9 expression reached the entire gonad at a time point, it was downregulated and became restricted to the central area in which testis cords were organized. MIS, a marker of Sertoli cells, appeared only in well-organized testis cords. We speculate that the SRY protein from the Y(TIR)-chromosome is inefficient in upregulating the Sox9 gene on the B6 background, allowing the initiation of ovarian differentiation.

Virtual breakout rooms: an effective approach to offer guidance to medical students on residency applications.

Peer mentorship on residency applications has been difficult due to recent public health measures, prompting a shift from in-person events to virtual platforms. To address gaps in career exploration, we created a virtual, non-recorded space that allowed medical students and residents to discuss the Canadian Resident Matching Service (CaRMS) process meaningfully and transparently. Attendees reported a greater understanding of the match process and reduced anxiety after the event. This model provides a virtual framework that can be adapted for various mentorship opportunities.

Le mentorat par les pairs offert aux étudiants concernant les demandes de résidence a été difficile à mettre en œuvre en raison des récentes mesures de santé publique, entraînant le transfert des activités en personne vers les plateformes virtuelles. Pour combler les lacunes dans l'exploration des carrières, nous avons créé un espace virtuel, sans enregistrement, qui a permis aux étudiants et aux résidents de discuter du processus du Service canadien de jumelage des résidents (CaRMS) de manière approfondie et transparente. Les participants ont indiqué qu'à la suite de la rencontre, ils comprenaient mieux le processus de jumelage et qu'ils étaient moins anxieux. Ce modèle virtuel peut être adapté et utilisé pour diverses activités de mentorat.

Factors Contributing to the Need for Non-Elective Explant following Surgical Repair of Tibial Tuberosity Avulsion Fracture.

OBJECTIVE: The aim of this study was to evaluate factors contributing to the need for non-elective explant following surgical repair of tibial tuberosity avulsion fractures. STUDY DESIGN: Retrospective multicentre case-control study. Over a 5-year period, dogs (n = 63) that underwent surgical repair of tibial tuberosity avulsion fractures (n = 64) were considered. Dogs that underwent a non-elective explant were compared with those that did not. Continuous variables were compared with the Mann-Whitney U test. Categorical variables were compared with the Fisher's exact test. Variables which were significant on univariate analysis were entered into a multiple logistic regression model. Significance was set at p < 0.05. RESULTS: Non-elective explant was performed in 20/64 fractures and elective explant was performed in 2/64 fractures. Neutered dogs were found to be 19 times (95% confidence interval: 2.1-172) more likely to require explant compared with intact dogs (p = 0.009). Every 0.25 mm increase in average pin size was found to make it 2.5 times (95% confidence interval: 1.3-4.9) more likely to require explant (p = 0.006). CONCLUSION: The findings suggest that use of the smallest appropriate pin should be considered for standard surgical repair of tibial tuberosity avulsion fractures to minimize the risk of requiring non-elective explant.

Clinical research: protection of the "vulnerable"?

In this age of evidence-based medicine, clinical research is critical for developing new therapeutics and determining the best way to use these therapies. To perform appropriate clinical research, researchers must adhere to ethical standards. These standards have developed in large part as a response to egregious violations of ethically appropriate behavior. In this respect certain populations have been identified as at risk of being treated inappropriately in medical research. Current ethical guidelines prohibit or severely limit what types of research can be performed involving these "vulnerable" populations. Although this might protect these populations, the lack of research on them might actually do harm in limiting their access to life-saving therapies. We explore the historical underpinnings of protecting the vulnerable populations and whether a newer ethical paradigm that would allow for protected research on these populations should be adopted by society.

Respiratory virus and asthma: the role of immunoglobulin E.

BACKGROUND: Atopic diseases, including asthma, have increased to epidemic proportions in the westernized world. The reasons for this increase are not known, nor are the mechanisms behind the development of these diseases. An interesting aspect of atopic disease is the role of respiratory viruses in the development of asthma and atopy. In fact, severe respiratory viral infection in infancy has been associated with a greatly increased risk of asthma. OBJECTIVE: This paper explores potential mechanisms through which viruses impart an increased risk of asthma, focusing on new pathways in mouse models of atopy. METHODS: A search of MEDLINE (1950-March 2008) was conducted using terms that included viral-induced wheeze, respiratory virus, asthma, IgE, and dendritic cells. RESULTS: A total of 1643 publications were identified that contained > or = 1 of the search terms; however, only 7 of these focused on immunoglobulin E (IgE) and the viral risk of asthma, and only 1 of the 7 explored the role of dendritic cells in this process. The latter study suggested a mechanistic link between lung dendritic cells and the development of postviral atopic disease. Important in this pathway is the generation of IgE, its high-affinity receptor, and the T-cell chemoattractant CCL28. CONCLUSIONS: Data from recent mouse models suggest that the development of asthma after severe respiratory viral infection may be the result of a response generated by production of antiviral IgE, which is capable of engaging dendritic cells to form a chemoattractant for interleukin-13-producing T cells. This new paradigm points to a focus for development of future therapies to prevent or at least ameliorate post- viral atopic disease.

Racial Differences in Mental Health Recovery among Veterans with Serious Mental Illness.

Black consumers with serious mental illness (SMI) face significant challenges in obtaining quality mental health care and are at risk for experiencing significant disparities in mental health care outcomes, including recovery from mental illness. Patient-provider interactions may partly contribute to this disparity. The purpose of the current study was to understand the effects of race, psychosis, and therapeutic alliance on mental health recovery orientation among Veterans with SMI. Participants were Veterans who had an SMI being treated at two Veteran Affairs outpatient mental health clinics by a psychiatrist or nurse practitioner. Participants completed the Behavior and Symptom Identification Scale (BASIS-24), Mental Health Recovery Measure, and patient-report Scale to Assess the Therapeutic Relationship (STAR-P) which includes three subscales: positive collaboration, positive clinician input, and non-supportive clinician input. Regression analyses were used to determine interactive effects of race, psychosis severity, and therapeutic alliance variables. The sample was 226 Veterans (50% black, 50% white). Black participants were slightly older (p < .05), had higher baseline psychosis (p < .05), higher mental health recovery (p < .05), and perceived less non-supportive clinician input (p < .01) than white participants. Regression analyses indicated a significant three-way interaction among race, psychosis, and positive collaboration (p < .01). Greater positive collaboration attenuated the negative effect of higher levels of psychosis on mental health recovery orientation for black participants. Conversely, for white participants, positive collaboration had little effect on the negative relationship between psychosis severity and mental health recovery orientation. Increased levels of psychosis may inhibit patients' perceptions of their ability to recover from SMI. However, for black participants, positive collaboration with mental health providers may moderate the effects of psychotic symptomatology.

Predictors of Employment Burnout Among VHA Peer Support Specialists.

OBJECTIVE: This study evaluated three domains of job burnout (emotional exhaustion, depersonalization, and personal accomplishment) and factors associated with burnout in a national sample of peer specialists (PSs) employed at 138 Veterans Health Administration (VHA) health care systems in 49 states. METHODS: Data were drawn from an observational study in which participants (N=152) completed online, self-report surveys about their mental health recovery, quality of life, and employment experiences at baseline, six months, and 12 months. Levels of burnout were analyzed at each time point, and regression analyses that controlled for baseline levels identified potential predictors of burnout (demographic, clinical, and employment characteristics) at six and 12 months. RESULTS: Compared with previously published burnout levels of other mental health workers in the VHA, PSs reported similar levels of emotional exhaustion, depersonalization, and personal accomplishment. At baseline, increased burnout was correlated with white race, fewer hours providing direct services, greater psychiatric symptoms, and lower self-efficacy. However, analyses did not reveal strong predictors of burnout scores at six or 12 months. CONCLUSIONS: In the first study to prospectively examine job burnout among PSs employed by the VHA, results illustrate the nuanced experience of burnout over a 12-month period and suggest the need for replication and further research on employment experiences of this emerging workforce.

The interactive effects of negative symptoms and social role functioning on suicide ideation in individuals with schizophrenia.

Findings regarding the protective effect of social role functioning on suicide ideation in individuals with schizophrenia have been mixed. One reason for such inconsistencies in the literature may be that individuals with prominent negative symptoms of schizophrenia may not experience a desire for social closeness, and therefore social role functioning may not influence suicide risk in these individuals. The aim of this study was to examine the moderating effects of self-reported desire for social closeness and interviewer-rated negative symptoms on the relationship between social role functioning and suicide ideation. Our sample consisted of 162 individuals who had been diagnosed with schizophrenia-spectrum disorders; all participants completed self-report questionnaires and clinician-administered interviews, and moderation hypotheses were tested with a non-parametric procedure. The results indicated that motivation and pleasure-related negative symptoms moderated the relationship between social role functioning and suicide ideation; self-reported desire for social closeness and negative symptoms related to expression did not have such a moderating effect. Specifically, better social role functioning was associated with less suicide ideation only in those individuals who had low motivation and pleasure-related negative symptoms; no significant relationship was observed between social role functioning and suicide ideation among those with elevated motivation and pleasure-related negative symptoms. These findings suggest that assessing for negative symptoms and social role functioning may inform suicide risk assessments in individuals with schizophrenia, and improving social role functioning may reduce suicide ideation among those with few motivation and pleasure-related negative symptoms.