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Introduction: In anticipation of institutional and community-wide COVID-19 immunization clinics, an educational program for the administration of COVID-19 vaccines was developed, collaborating with the Virginia Department of Health to train doctor of medicine and physician assistant students to serve as vaccinators. Faculty and students also worked with state legislatures to propose and enact a bill that would enable such students to vaccinate patients. Methods: Between January 2021 and August 2022, 263 student volunteers completed 3,685 person-hours, administering 48,279 doses. On the basis of community need, the majority of vaccines were administered at mass vaccination clinics in Chesapeake (47%) and Norfolk (22%) in Virginia. One year after the first COVID-19 immunization clinic, the authors surveyed students who assisted with clinics, utilizing a Likert scale and free-text responses to elicit feedback about the training and volunteer experience. Results: Volunteers ranked the vaccination clinics among the top third of established volunteer experiences offered at Eastern Virginia Medical School, and 75% of respondents believed that this training should be a permanent part of their program curriculum. Conclusions: This paper presents a strategy for preparing students to serve as vaccinators during public health emergencies and show how other institutions of medical education can prepare for and engage student participation in vaccination campaigns and emergent health initiatives.

Streptococcus pyogenes, also known as group A Streptococcus (GAS), is a ubiquitous pathogen that produces an array of human disease, including focal infections (e.g., pharyngitis, pyoderma) with or without localized [...]

IntroductionPancreatic ductal adenocarcinoma (PDAC) is a lethal cancer associated with an immunosuppressive environment. Neutrophil extracellular traps (NETs) were initially described in the context of infection but have more recently been implicated in contributing to the tolerogenic immune response in PDAC. Thus, NETs are an attractive target for new therapeutic strategies. Group A Streptococcus (GAS) has developed defensive strategies to inhibit NETs.MethodsIn the present work, we propose utilizing intra-tumoral GAS injection to stimulate anti-tumor activity by inhibiting cancer-promoting NETs. Mice harboring Panc02 or KPC subcutaneous tumors injected with three different M-type GAS strains. Tumors and spleens were harvested at the endpoint of the experiments to assess bacterial colonization and systemic spread, while sera were analyzed for humoral responses toward the streptococcal antigens, especially the M1 and Scl1 proteins. Role of the streptococcal collagen-like protein 1 (Scl1) in anti-PDAC activity was assessed in vivo after intratumoral injection with M1 GAS wild-type, an isogenic mutant strain devoid of Scl1, or a complemented mutant strain with restored scl1 expression. In addition, recombinant Scl1 proteins were tested for NET inhibition using in vitro and ex vivo assays assessing NET production and myeloperoxidase activity.ResultsInjection of three different M-type GAS strains reduced subcutaneous pancreatic tumor volume compared to control in two different murine PDAC models. Limitation of tumor growth was dependent on Scl1, as isogenic mutant strain devoid of Scl1 did not reduce tumor size. We further show that Scl1 plays a role in localizing GAS to the tumor site, thereby limiting the systemic spread of bacteria and off-target effects. While mice did elicit a humoral immune response to GAS antigens, tested sera were weakly immunogenic toward Scl1 antigen following intra-tumoral treatment with Scl1-expressing GAS. M1 GAS inhibited NET formation when co-cultured with neutrophils while Scl1-devoid mutant strain did not. Recombinant Scl1 protein inhibited NETs ex vivo in a dose-dependent manner by suppressing myeloperoxidase activity.DiscussionAltogether, we demonstrate that intra-tumoral GAS injections reduce PDAC growth, which is facilitated by Scl1, in part through inhibition of cancer promoting NETs. This work offers a novel strategy by which NETs can be targeted through Scl1 protein and potentiates its use as a cancer therapeutic.

Clindamycin and β-lactam antibiotics have been mainstays for treating invasive group A Streptococcus (iGAS) infection, yet such regimens might be limited for strains displaying MLSB phenotypes. We investigated 76 iGAS isolates from 66 patients in West Virginia, USA, during 2020–2021. We performed emm typing using Centers for Disease Control and Prevention guidelines and assessed resistance both genotypically and phenotypically. Median patient age was 42 (range 23–86) years. We found 76% of isolates were simultaneously resistant to erythromycin and clindamycin, including all emm92 and emm11 isolates. Macrolide resistance was conferred by the plasmid-borne ermT gene in all emm92 isolates and by chromosomally encoded ermA, ermB, and a single mefA in other emm types. Macrolide-resistant iGAS isolates were typically resistant to tetracycline and aminoglycosides. Vulnerability to infection was associated with socioeconomic status. Our results show a predominance of macrolide-resistant isolates and a shift in emm type distribution compared with historical reports.

Pancreatic cancer is a highly aggressive form of cancer with a five-year survival rate of only ten percent. Pancreatic ductal adenocarcinoma (PDAC) accounts for ninety percent of those cases. PDAC is associated with a dense stroma that confers resistance to current treatment modalities. Increasing resistance to cancer treatments poses a challenge and a need for alternative therapies. Bacterial mediated cancer therapies were proposed in the late 1800s by Dr. William Coley when he injected osteosarcoma patients with live streptococci or a fabrication of heat-killed Streptococcus pyogenes and Serratia marcescens known as Coley’s toxin. Since then, several bacteria have gained recognition for possible roles in potentiating treatment response, enhancing anti-tumor immunity, and alleviating adverse effects to standard treatment options. This review highlights key bacterial mechanisms and structures that promote anti-tumor immunity, challenges and risks associated with bacterial mediated cancer therapies, and applications and opportunities for use in PDAC management.

Burkholderia pseudomallei is a gram-negative bacterium that is the etiological agent of the tropical disease melioidosis. Currently, there is no licensed vaccine for melioidosis, but numerous candidates are being tested for protective efficacy and characterization of the elicited immune response. Our lab has previously reported the immunogenicity of a Bucl8-protein-based peptide antigen, designated L1-CRM197 (Cross-reacting material 197). When given subcutaneously, this vaccine formulation promoted a strong Th2 (IgG1) antibody response, however immunization did not protect from death. In this study, we hypothesized that an intranasally administered L1-CRM197 vaccine would induce protective mucosal immunity. To evaluate vaccine efficacy, we developed a surrogate Burkholderia infection model that employs outbred CD-1 mice which imitates the immunogenetic diversity of humans. Mice were immunized with either L1-CRM197 adjuvanted with fluorinated cyclic diguanosine monophosphate (FCDG) or with FCDG-only control. These mice were then challenged intranasally with an infectious dose of a luminescent strain of B. thailandensis E264 two weeks post-immunization, and correlates of protection were assessed in euthanized mice on days 1, 2, 3, and 7 post-infection. Overall, intranasal vaccination, compared to subcutaneous administration, induced a stronger Th1 (IgG2a/2b) to Th2 (IgG1) antibody response and promoted anti-L1 nasal, pulmonary, and systemic IgA. Additionally, sera IgG from L1-CRM197-vaccinated mice recognized whole-cell B. thailandensis and B. pseudomallei, a select agent exempt strain Bp82. Vaccination ameliorated disease indicators, including luminescent signal and bacterial cell counts, weight and temperature loss, and organ weight, which negatively correlated with IgG2a antibody levels and mucosa-stimulating cytokines IL-13 and IL-9. L1-CRM197-vaccinated mice also had earlier resolution of inflammatory and tissue-damaging cytokines compared to the FCDG-only controls. These results suggest a balanced humoral and cell-mediated response, along with mucosa-based immunity are beneficial for protection. Future efforts should further assess mucosal cellular and humoral mechanisms of protection and test such protection, using aerosolized B. pseudomallei select agent strain(s).

Wastewater-based epidemiology is being used as a tool to monitor the spread of COVID-19 and provide an early warning for the presence or increase of clinical cases in a community. The majority of wastewater-based epidemiology for COVID-19 tracking has been utilized in sewersheds that service populations in the tens-to-hundreds of thousands. Few studies have been conducted to assess the usefulness of wastewater in predicting COVID-19 clinical cases specifically in rural areas. This study collected samples from 16 locations across the Eastern Upper Peninsula of Michigan from June to December 2021. Sampling locations included 12 rural municipalities, a Tribal housing community and casino, a public university, three municipalities that also contained a prison, and a small island with heavy tourist traffic. Samples were analyzed for SARS-CoV-2 N1, N2, and variant gene copies using reverse transcriptase droplet digital polymerase chain reaction (RT-ddPCR). Wastewater N1 and N2 gene copies and clinical case counts were correlated to determine if wastewater results were predictive of clinical cases. Significant correlation between N1 and N2 gene copies and clinical cases was found for all sites (⍴= 0.89 to 0.48). N1 and N2 wastewater results were predictive of clinical case trends within 0-7 days. The Delta variant was detected in the Pickford and St. Ignace samples more than 12-days prior to the first reported Delta clinical cases in their respective counties. Locations with low correlation could be attributed to their high rates of tourism. This is further supported by the high correlation seen in the public university, which is a closed population. Long-term wastewater monitoring over a large, rural geographic area is useful for informing the public of potential outbreaks in the community regardless of asymptomatic cases and access to clinical testing.

Altered motor and social-communicative abilities in infancy have been linked to later ASD diagnosis. Most diagnostic instruments for ASD cannot be utilized until 12 months, and the average child is diagnosed substantially later. Imitation combines motor and social-communicative skills and is commonly atypical in infants at risk for ASD. However, few measures have been developed to assess infant imitation clinically. One barrier to the diagnostic age gap of ASD is accessibility of screening and diagnostic services. Utilization of caregiver report to reliably screen for ASD mitigates such barriers and could aid in earlier detection. The present study developed and validated a caregiver-report measure of infant imitation at 4, 6, and 9 months and explored the relationship between caregiver-reported imitation and motor abilities with later ASD risk. Participants (N = 571) were caregivers of term and preterm infants recruited as part of a large multi-site study of PediaTrac™, a web-based tool for monitoring and tracking infant development. Caregivers completed online surveys and established questionnaires on a schedule corresponding to well-child visits from birth to 18 months, including the M-CHAT-R/F at 18 months. Distinct imitation factors were derived from PediaTrac at 4, 6, and 9 months via factor analysis. The results supported validity of the imitation factors via associations with measures of infant communication (CSBS; ASQ). Imitation and motor skills at 9 months predicted 18-month ASD risk over and above gestational age. Implications for assessment of infant imitation, detecting ASD risk in the first year, and contributing to access to care are discussed. [ABSTRACT FROM AUTHOR]

Caregiver report is the most feasible way to assess early childhood development but is susceptible to the influences of response style and sociodemographic factors. In a sample of 571 caregiver-infant dyads (47.8% female; 48% White), we compared caregiver reports on the Ages and Stages Questionnaire-Third Edition (ASQ-3) with reports on a novel, web-based assessment, PediaTrac™. Ratings on PediaTrac correlated with ratings on the ASQ-3 at all time points (2, 4, 6, and 9 months). Caregiver age, response style, and sociodemographic factors accounted for significant variance on both measures. Developmental reporting of early childhood skills is influenced by caregiver response style and sociodemographic factors. These influences must be considered in order to ensure the accurate identification of infant developmental status.

The primary aim of this study was to evaluate the convergent validity of the Motor domain (MOT) of PediaTracTM v3.0, an online developmental tracking instrument based on caregiver reports, with fine and gross motor domains (ASQ-FM and ASQ-GM) of the Ages and Stages Questionnaire (ASQ-3) in infants between 2- and 9 months of age. Participants were caregivers of 571 infants born term or preterm (gestational age <37 weeks) enrolled in a multi-site psychometric study of PediaTracTM. Findings revealed significant correlations between MOT and ASQ-3 scores at 2, 4, 6, and 9 months across time periods, term-preterm status, and biological sex. A significantly higher percentage of infants born preterm, compared with those born at term, was identified as a moderate or high risk on both the ASQ-3 and PediaTrac. Future investigations are warranted to further examine the psychometric properties of the MOT domain, including sensitivity, specificity, and positive and negative predictive value. [ABSTRACT FROM AUTHOR]

Simple Summary: The prediction of tumor response after neoadjuvant FLOT therapy is highly necessary. The use of deep learning on gastroesophageal biopsies enabled us to extract predictive information. This prediction model could be easily applied in clinical decision making. Patients could avoid unnecessary treatment or receive an intensified FLOT therapy. Background: The aim of this study was to establish a deep learning prediction model for neoadjuvant FLOT chemotherapy response. The neural network utilized clinical data and visual information from whole-slide images (WSIs) of therapy-naïve gastroesophageal cancer biopsies. Methods: This study included 78 patients from the University Hospital of Cologne and 59 patients from the University Hospital of Heidelberg used as external validation. Results: After surgical resection, 33 patients from Cologne (42.3%) were ypN0 and 45 patients (57.7%) were ypN+, while 23 patients from Heidelberg (39.0%) were ypN0 and 36 patients (61.0%) were ypN+ (p = 0.695). The neural network had an accuracy of 92.1% to predict lymph node metastasis and the area under the curve (AUC) was 0.726. A total of 43 patients from Cologne (55.1%) had less than 50% residual vital tumor (RVT) compared to 34 patients from Heidelberg (57.6%, p = 0.955). The model was able to predict tumor regression with an error of ±14.1% and an AUC of 0.648. Conclusions: This study demonstrates that visual features extracted by deep learning from therapy-naïve biopsies of gastroesophageal adenocarcinomas correlate with positive lymph nodes and tumor regression. The results will be confirmed in prospective studies to achieve early allocation of patients to the most promising treatment. [ABSTRACT FROM AUTHOR]

Background: Adverse airway events (AAEs) are rare but devastating complications following palatoplasty. The purpose of this study is to evaluate patient risk factors for their effect on these complications. We hypothesize that prolonged operative time and the presence of multiple medical comorbidities are risk factors for AAEs. Design: Retrospective cohort study. Setting: Participant hospitals in the Pediatric American College of Surgeons National Surgical Quality Improvement Program year 2016–2019. Patients: Cases of palatoplasty in children under 3 years of age. Outcomes: Adverse airway events including postoperative reintubation or any requirement of postoperative mechanical ventilation. Results: A total of 6668 patients met inclusion criteria. The median operative time was 126 min (IQR 82). AAEs were identified in 107 (1.6%) patients. The incidence of risk factors was found to increase with age and AAEs were more prevalent in younger and older patients. Although patients in the older age groups had significantly higher burden of comorbidities, differences in age were not independently associated with AAEs. Following multivariable logistic regressions, operative times greater than 2 h, ASA class ≥3, >3 medical comorbidities, and black race were found to be significant independent risk factors. Conclusions: In this large, retrospective database study in palatoplasty, increased operative time, ASA classification ≥3, multiple comorbidities, and black race were independently associated with AAEs. [ABSTRACT FROM AUTHOR]

Aim: To examine the extent to which estimates of a latent trait or underlying construct of motor ability differ in infants born at term and preterm, based on caregiver ratings of the motor domain of PediaTrac v3.0. Method: The sample consisted of 571 caregiver–infant dyads (331 born at term, 240 born preterm), 48% female, with 51.7% of caregivers identifying as an ethnic minority. Latent trait of motor ability was estimated based on item response theory modeling. Gestational group differences (term and preterm birth) were examined at the newborn/term‐equivalent, 2‐, 4‐, 6‐, 9‐, and 12‐month time points. Results: Caregiver ratings of latent trait of motor ability were reliably modeled across the range of abilities at each time point. While the group born preterm exhibited significantly more advanced motor abilities at the term‐equivalent time point, by 6 months the group born at term was more advanced. Biological sex difference main and interaction effects were not significant. Interpretation: Caregivers provided reliable, longitudinal estimates of motor ability in infancy, reflecting important differences in the motor development of infants born at term and preterm. The findings suggest that significant motor development occurs in infants born preterm from birth to the term‐equivalent time point and provide a foundation to examine motor growth trajectories as potential predictors in the early identification of neurodevelopmental conditions and needs. What this paper adds: Longitudinal caregiver ratings of motor function in early infancy yielded reliable estimates of the latent trait of motor ability.Motor ability at the term‐equivalent time point was higher in infants born preterm than infants born at term. [ABSTRACT FROM AUTHOR]

Bacterial efflux pumps are an important pathogenicity trait because they extrude a variety of xenobiotics. Our laboratory previously identified in silico Burkholderia collagen-like protein 8 (Bucl8) in the hazardous pathogens Burkholderia pseudomallei and Burkholderia mallei. We hypothesize that Bucl8, which contains two predicted tandem outer membrane efflux pump domains, is a component of a putative efflux pump. Unique to Bucl8, as compared to other outer membrane proteins, is the presence of an extended extracellular region containing a collagen-like (CL) domain and a non-collagenous C-terminus (Ct). Molecular modeling and circular dichroism spectroscopy with a recombinant protein, corresponding to this extracellular CL-Ct portion of Bucl8, demonstrated that it adopts a collagen triple helix, whereas functional assays screening for Bucl8 ligands identified binding to fibrinogen. Bioinformatic analysis of the bucl8 gene locus revealed it resembles a classical efflux-pump operon. The bucl8 gene is co-localized with downstream fusCDE genes encoding fusaric acid (FA) resistance, and with an upstream gene, designated as fusR, encoding a LysR-type transcriptional regulator. Using reverse transcriptase (RT)-qPCR, we defined the boundaries and transcriptional organization of the fusR-bucl8-fusCDE operon. We found exogenous FA induced bucl8 transcription over 80-fold in B. pseudomallei, while deletion of the entire bucl8 locus decreased the minimum inhibitory concentration of FA 4-fold in its isogenic mutant. We furthermore showed that the putative Bucl8-associated pump expressed in the heterologous Escherichia coli host confers FA resistance. On the contrary, the Bucl8-associated pump did not confer resistance to a panel of clinically-relevant antimicrobials in Burkholderia and E. coli. We finally demonstrated that deletion of the bucl8-locus drastically affects the growth of the mutant in L-broth. We determined that Bucl8 is a component of a novel tetrapartite efflux pump, which confers FA resistance, fibrinogen binding, and optimal growth.