Influenza A viral burst size from thousands of infected single cells using droplet quantitative PCR (dqPCR).

An important aspect of how viruses spread and infect is the viral burst size, or the number of new viruses produced by each infected cell. Surprisingly, this value remains poorly characterized for influenza A virus (IAV), commonly known as the flu. In this study, we screened tens of thousands of cells using a microfluidic method called droplet quantitative PCR (dqPCR). The high-throughput capability of dqPCR enabled the measurement of a large population of infected cells producing progeny virus. By measuring the fully assembled and successfully released viruses from these infected cells, we discover that the viral burst sizes for both the seasonal H3N2 and the 2009 pandemic H1N1 strains vary significantly, with H3N2 ranging from 101 to 104 viruses per cell, and H1N1 ranging from 101 to 103 viruses per cell. Some infected cells produce average numbers of new viruses, while others generate extensive number of viruses. In fact, we find that only 10% of the single-cell infections are responsible for creating a significant portion of all the viruses. This small fraction produced approximately 60% of new viruses for H3N2 and 40% for H1N1. On average, each infected cell of the H3N2 flu strain produced 709 new viruses, whereas for H1N1, each infected cell produced 358 viruses. This novel method reveals insights into the flu virus and can lead to improved strategies for managing and preventing the spread of viruses.

Novel Skin Prep Technique Reduces OR Preparation Times in a Randomized Trial for Podiatric and Orthopedic Procedures.

The primary objectives of any high-volume surgery department should be patient safety, block time utilization and operating room efficiency. Reducing preparation time in the OR prior to actual surgery can improve operating room efficiency and utilization, but only if patient safety can be maintained. With this goal, this study evaluated a novel skin preparation technique using a device named ULTRAPREP™, a sterile, medical-grade plastic bag that is applied to the upper or lower extremity in the pre-operative holding area which allows for skin disinfection outside the OR (referred to as "disinfection bag"). The study compared preparation times required in the OR and antiseptic efficiency (through Colony Forming Units (CFU) counts) for traditional methods versus using the disinfection bag on a total of 115 patients undergoing podiatric or orthopedic surgeries (upper and lower extremities) in one hospital. The disinfection bag reduced skin preparation time in the OR from 16.8±3.5min to 10.9±2.7min, which was a 35.2% reduction, and was statistically significant (p<0.01). Skin antisepsis met safety standards of <15 CFUs for all cases regardless of preparation type at 48h and 72h. There was no statistical difference in CFU levels between the traditional and disinfection bag methods at 48h or 72h (p>0.11). Therefore, ULTRAPREP™ has shown the ability to decrease operating room time while keeping surgical site infection rates to a minimum. Minimizing activities in the OR optimizes use of this costly resource and brings overall savings to the surgery department.

Craving among patients seeking treatment for substance use disorder.

BACKGROUND: Craving has been implicated as a central feature of addiction and a predictor of relapse. However, a complete understanding of how craving varies across patient populations is lacking. This study aimed to better inform the effective and accurate use of craving as a clinical prognostic tool for patients with substance use disorders (SUD). METHODS: This cross-sectional study utilized information gathered on patients (n = 112) entering specialty treatment for a SUD. Craving in the prior 30 days was assessed with a single item with other intake questionnaires. RESULTS: Patients who reported substance use in the last 30 days were more likely to report craving compared to patients who did not report substance use in the last 30 days (AOR = 6.86 [95% CI 2.17-21.7], p-value = 0.001). Patients who reported Hispanic/Latinx ethnicity were less likely to report craving compared to patients who did not report Hispanic/Latinx ethnicity (AOR = 0.28 [95% CI 0.08-0.95], p-value = 0.04). There was no association between craving and Adverse Childhood Events (OR = 1.03 [95% CI 0.84-1.25], p-value = 0.81). CONCLUSION: The association between recent substance use and craving supports previous findings. The observed variation in craving among patients who report Hispanic/Latinx ethnicity is novel and suggests socio-cultural influences and possibly genetic factors influencing reported craving amongst patients. Additional research is needed to further understand the underlying factors leading to this finding, in order for better utilization of craving as a clinical indicator across patient populations.

A Quantitative Method to Measure the Kinetics of Elemental Mercury Emissions From Black Shale (Nova Scotia, Canada).

A controlled chamber method using continuous gold trap atomic fluorescence spectroscopy (AFS) (Tekran 2537X) for the analysis of Hg(0) emissions from moderate mass rock samples was developed and tested. A series of black shale and other bedrock samples from Nova Scotia, Canada, were used to test the method and its reproducibility. Hg(0) emissions at 170°C were measured to quantify both free surficial Hg(0) and Hg(0) that had penetrated the rock structure. High volumes of chamber air (45 L) were sampled using 30 min collection times to achieve detectable elemental mercury (Hg(0)) emissions. We found higher percentage masses of Hg(0) were released (1.1%-4.1% of total Hg mass present) in black shale samples as compared to granite and basalt samples from the same region (0.0%-0.3% released) over 350 h of continuous analysis time. The pseudo first order emission rate constants ranged from 0.015-0.245 h-1 (mean 0.063 h-1, standard deviation (SD) 0.102) for the black shale samples analyzed and was 0.004 h-1 for the granite sample. The 24-h zero-order emission rate constants ranged between 0.41 and 3.54 ng h-1 (mean 1.4 ng h-1, SD 1.3) for the black shale samples analyzed and were ~ 0.01 ng h-1 for the granite and basalt samples. This technique has useful implications for examining rock properties and Hg(0) emission rates.

Lgr5-expressing secretory cells form a Wnt inhibitory niche in cartilage critical for chondrocyte identity.

Osteoarthritis is a degenerative joint disease that causes pain, degradation, and dysfunction. Excessive canonical Wnt signaling in osteoarthritis contributes to chondrocyte phenotypic instability and loss of cartilage homeostasis; however, the regulatory niche is unknown. Using the temporomandibular joint as a model in multiple species, we identify Lgr5-expressing secretory cells as forming a Wnt inhibitory niche that instruct Wnt-inactive chondroprogenitors to form the nascent synovial joint and regulate chondrocyte lineage and identity. Lgr5 ablation or suppression during joint development, aging, or osteoarthritis results in depletion of Wnt-inactive chondroprogenitors and a surge of Wnt-activated, phenotypically unstable chondrocytes with osteoblast-like properties. We recapitulate the cartilage niche and create StemJEL, an injectable hydrogel therapy combining hyaluronic acid and sclerostin. Local delivery of StemJEL to post-traumatic osteoarthritic jaw and knee joints in rabbit, rat, and mini-pig models restores cartilage homeostasis, chondrocyte identity, and joint function. We provide proof of principal that StemJEL preserves the chondrocyte niche and alleviates osteoarthritis.

Integrating EM and Patch-seq data: Synaptic connectivity and target specificity of predicted Sst transcriptomic types.

Neural circuit function is shaped both by the cell types that comprise the circuit and the connections between those cell types 1 . Neural cell types have previously been defined by morphology 2, 3 , electrophysiology 4, 5 , transcriptomic expression 6-8 , connectivity 9-13 , or even a combination of such modalities 14-16 . More recently, the Patch-seq technique has enabled the characterization of morphology (M), electrophysiology (E), and transcriptomic (T) properties from individual cells 17-20 . Using this technique, these properties were integrated to define 28, inhibitory multimodal, MET-types in mouse primary visual cortex 21 . It is unknown how these MET-types connect within the broader cortical circuitry however. Here we show that we can predict the MET-type identity of inhibitory cells within a large-scale electron microscopy (EM) dataset and these MET-types have distinct ultrastructural features and synapse connectivity patterns. We found that EM Martinotti cells, a well defined morphological cell type 22, 23 known to be Somatostatin positive (Sst+) 24, 25 , were successfully predicted to belong to Sst+ MET-types. Each identified MET-type had distinct axon myelination patterns and synapsed onto specific excitatory targets. Our results demonstrate that morphological features can be used to link cell type identities across imaging modalities, which enables further comparison of connectivity in relation to transcriptomic or electrophysiological properties. Furthermore, our results show that MET-types have distinct connectivity patterns, supporting the use of MET-types and connectivity to meaningfully define cell types.

Mutational analysis of a conserved positive charge in the c-ring of E. coli ATP synthase.

F1Fo ATP synthase is a ubiquitous molecular motor that utilizes a rotary mechanism to synthesize adenosine triphosphate (ATP), the fundamental energy currency of life. The membrane-embedded Fo motor converts the electrochemical gradient of protons into rotation, which is then used to drive the conformational changes in the soluble F1 motor that catalyze ATP synthesis. In E. coli, the Fo motor is composed of a c10 ring (rotor) alongside subunit a (stator), which together provide two aqueous half channels that facilitate proton translocation. Previous work has suggested that Arg50 and Thr51 on the cytoplasmic side of each subunit c are involved in the proton translocation process, and positive charge is conserved in this region of subunit c. To further investigate the role of these residues and the chemical requirements for activity at these positions, we generated 13 substitution mutants and assayed their in vitro ATP synthesis, H+ pumping, and passive H+ permeability activities, as well as the ability of mutants to carry out oxidative phosphorylation in vivo. While polar and hydrophobic mutations were generally tolerated in either position, introduction of negative charge or removal of polarity caused a substantial defect. We discuss the possible effects of altered electrostatics on the interaction between the rotor and stator, water structure in the aqueous channel, and interaction of the rotor with cardiolipin.

Championing inclusive terminology in ecology and evolution.

Amid a growing disciplinary commitment to inclusion in ecology and evolutionary biology (EEB), it is critical to consider how the use of scientific language can harm members of our research community. Here, we outline a path for identifying and revising harmful terminology to foster inclusion in EEB.

Respiratory viruses: New frontiers-a Keystone Symposia report.

Respiratory viruses are a common cause of morbidity and mortality around the world. Viruses like influenza, RSV, and most recently SARS-CoV-2 can rapidly spread through a population, causing acute infection and, in vulnerable populations, severe or chronic disease. Developing effective treatment and prevention strategies often becomes a race against ever-evolving viruses that develop resistance, leaving therapy efficacy either short-lived or relevant for specific viral strains. On June 29 to July 2, 2022, researchers met for the Keystone symposium "Respiratory Viruses: New Frontiers." Researchers presented new insights into viral biology and virus-host interactions to understand the mechanisms of disease and identify novel treatment and prevention approaches that are effective, durable, and broad.

Using teledermatology to prevent and diagnose skin cancer in the rural United States.

ABSTRACT: In the United States, skin cancer is a prevalent and sometimes preventable form of cancer that causes a significant disease burden in rural and urban communities. Studies have shown, however, that rural residents are less likely to engage in primary prevention behaviors against skin cancer, and rural populations have higher skin cancer incidence and mortality than urban residents. Teledermatology can be used to address disparities in both skin cancer diagnosis and health education to improve dermatology outcomes in rural communities. This article reviews teledermatology formats, barriers, and benefits to encourage integration of teledermatology modalities into clinical practice.

Single-Cell Infection of Influenza A Virus Using Drop-Based Microfluidics.

Drop-based microfluidics has revolutionized single-cell studies and can be applied toward analyzing tens of thousands to millions of single cells and their products contained within picoliter-sized drops. Drop-based microfluidics can shed insight into single-cell virology, enabling higher-resolution analysis of cellular and viral heterogeneity during viral infection. In this work, individual A549, MDCK, and siat7e cells were infected with influenza A virus (IAV) and encapsulated into 100-µm-size drops. Initial studies of uninfected cells encapsulated in drops demonstrated high cell viability and drop stability. Cell viability of uninfected cells in the drops remained above 75%, and the average drop radii changed by less than 3% following cell encapsulation and incubation over 24 h. Infection parameters were analyzed over 24 h from individually infected cells in drops. The number of IAV viral genomes and infectious viruses released from A549 and MDCK cells in drops was not significantly different from bulk infection as measured by reverse transcriptase quantitative PCR (RT-qPCR) and plaque assay. The application of drop-based microfluidics in this work expands the capacity to propagate IAV viruses and perform high-throughput analyses of individually infected cells. IMPORTANCE Drop-based microfluidics is a cutting-edge tool in single-cell research. Here, we used drop-based microfluidics to encapsulate thousands of individual cells infected with influenza A virus within picoliter-sized drops. Drop stability, cell loading, and cell viability were quantified from three different cell lines that support influenza A virus propagation. Similar levels of viral progeny as determined by RT-qPCR and plaque assay were observed from encapsulated cells in drops compared to bulk culture. This approach enables the ability to propagate influenza A virus from encapsulated cells, allowing for future high-throughput analysis of single host cell interactions in isolated microenvironments over the course of the viral life cycle.

Effects of rumen-native microbial feed supplementation on milk yield, composition, and feed efficiency in lactating dairy cows.

The objective of this study was to evaluate the effects of two rumen-native microbial feed supplements (MFS) on milk production, milk composition, and feed efficiency. A total of 90 multiparous cows between 40 and 60 d in milk were enrolled in a randomized block design study. Within each block (baseline milk yield), cows were randomly assigned to: control (no microbial feed supplementation), MFS1 (0.33 g/kg total mixed ration [TMR] of an MFS containing a minimum of Clostridium beijerinckii at 2 × 106 CFU/g and Pichia kudriavzevii at 2 × 107 CFU/g), or MFS2 (0.33 g/kg TMR of a MFS containing a minimum of C. beijerinckii at 2 × 106 CFU/g, P. kudriavzevii at 2 × 107 CFU/g, Ruminococcus bovis at 2 × 107 CFU/g, and Butyrivibrio fibrisolvens at 2 × 107 CFU/g). Cows were housed in a single group and fed the study diets ad libitum for 270 d. Individual milk yield was recorded using electronic milk meters, and milk fat and protein were measured using optical in-line analyzers at each of two daily milkings. Treatment and treatment by time effects were assessed through multiple linear regression analyses. Treatment effects were observed for milk and energy-corrected milk (ECM) yields, milk fat and protein yields and concentrations, dry matter intake (DMI), and feed efficiency; those effects were conditional to time for milk yield, DMI, and feed efficiency. Overall, milk, ECM, fat, and protein yields were higher for MFS2 compared with control cows (+3.0, 3.7, 0.12, and 0.12 kg/d, respectively). Compared with MFS1, milk yield was higher and protein yield tended to be higher for MFS2 cows (+2.9 and 0.09 kg/d, respectively). In contrast, MFS1 cows produced 0.17 and 0.08 units of percentage per day more fat and protein than MFS2 cows, and 0.07 units of percentage per day more protein than control cows. Dry matter intake and feed efficiency were higher for MFS2 cows compared with MFS1 cows (+1.3 kg/d and 0.06, respectively), and feed efficiency was higher for MFS2 cows compared with control cows (+0.04). Where observed, treatment by time effects suggest that the effects of MFS2 were more evident as time progressed after supplementation was initiated. No effects of microbial supplementation were observed on body weight, body condition score, somatic cell count, or clinical mastitis case incidence. In conclusion, the supplementation of MFS2 effectively improved economically important outcomes such as milk yield, solids, and feed efficiency.

This study evaluates the effects of two rumen-native microbial feed supplements (MFS) on milk yield, composition, and feed efficiency in lactating dairy cows. Ninety multiparous Holstein cows between 40 and 60 d in milk were assigned to control (no microbial feed supplementation), MFS1 (Clostridium beijerinckii and Pichia kudriavzevii), or MFS2 (C. beijerinckii, P. kudriavzevii, Ruminococcus bovis, and Butyrivibrio fibrisolvens) total mixed ration supplementation. Overall, MFS2 cows had higher milk and milk component yields than control and MFS1, while MFS1 cows had higher milk component concentrations than control and MFS2. Feed efficiency was higher for MFS2 compared with control and MFS1 cows. Microbial feed supplementation improved economically important outcomes such as milk yield, solids, and feed efficiency.

Open set classification strategies for long-term environmental field recordings for bird species recognition.

Deep learning is one established tool for carrying out classification tasks on complex, multi-dimensional data. Since audio recordings contain a frequency and temporal component, long-term monitoring of bioacoustics recordings is made more feasible with these computational frameworks. Unfortunately, these neural networks are rarely designed for the task of open set classification in which examples belonging to the training classes must not only be correctly classified but also crucially separated from any spurious or unknown classes. To combat this reliance on closed set classifiers which are singularly inappropriate for monitoring applications in which many non-relevant sounds are likely to be encountered, the performance of several open set classification frameworks is compared on environmental audio datasets recorded and published within this work, containing both biological and anthropogenic sounds. The inference-based open set classification techniques include prediction score thresholding, distance-based thresholding, and OpenMax. Each open set classification technique is evaluated under multi-, single-, and cross-corpus scenarios for two different types of unknown data, configured to highlight common challenges inherent to real-world classification tasks. The performance of each method is highly dependent upon the degree of similarity between the training, testing, and unknown domain.

Sea ice extent and phenology influence breeding of high-Arctic seabirds: 4 decades of monitoring in Nunavut, Canada.

Seabirds breeding in the high Arctic contend with variable annual sea ice conditions, with important consequences depending on a species' unique reproductive and foraging ecology. We assessed the influence of sea ice extent and phenology on seabird breeding biology using monitoring data collected for northern fulmar (Fulmarus glacialis), glaucous gull (Larus hyperboreus), black-legged kittiwake (Rissa tridactyla), and thick-billed murre (Uria lomvia) breeding at Prince Leopold Island, Nunavut, Canada over 4 decades. We expected that years of later sea ice break-up and greater ice cover around the colony would create greater challenges to foraging and could result in delayed nest initiation, decreased colony attendance, and lower nesting success, but with distinct responses from each species. We also tested for time-lagged effects of ice conditions, where sea ice in a given year could impact food availability or juvenile recruitment in later years. Ice conditions around the colony exhibited no significant overall temporal trends or changepoints over the past 50 years (1970-2021), while counts of kittiwakes and murres increased over the study period 1975-2013. No trends were evident in counts of fulmars or gulls or in egg-laying dates or nest success for any species. However, three species (all but glaucous gulls) exhibited unique responses between breeding metrics and sea ice, highlighting how breeding decisions and outcomes may differ among species under the same environmental conditions in a given year. Time-lagged effects were only detected for kittiwake nest counts, where the date of spring ice break-up around the colony was negatively associated with counts at a 5-year lag. Greater distances to open water were associated with lower colony attendance by fulmars and later nest initiation by kittiwakes and murres. Our analyses provide additional insights to effects of sea ice on high-Arctic seabird breeding ecology, which will be useful in predicting and planning for the complex effects of a changing climate and changing human pressures on this high-latitude ecosystem and for the management of high-Arctic marine-protected areas.

Utilizing Supplemental Online Modules for Physician Assistant Student Electrocardiogram Interpretation Training.

PURPOSE: The literature suggests that graduating medical and physician assistant (PA) students lack competency in electrocardiogram (ECG) interpretation. This project aimed to determine whether use of perceptual adaptive learning modules (PALMs) would improve PA students' ECG interpretation, alter self-perceptions of their ECG education, or both. METHODS: PALMs were incorporated into the PA curriculum after lecture-based ECG learning. Students' pretest, posttest, and delayed-posttest scores were then compared. Students' ability to correctly interpret ECGs (accuracy) and the percentage of ECGs accurately interpreted within 15 seconds or less (fluency) also were evaluated. Finally, students' perceptions of PALMs and overall ECG training were assessed. RESULTS: PALM training improved ECG interpretation accuracy and fluency (p < .0001), as well as delayed-posttest accuracy and fluency (p < .0001). The majority of student respondents felt supplemental training enhanced their learning. CONCLUSION: These perception results combined with data on ECG interpretation improvement supports continued use of supplemental PALMs in ECG interpretation training.

Total mercury, methylmercury, phosphate, and sulfate inputs to a bog ecosystem from herring gull (Larus smithsoniansus) guano.

Avian biovector transport is an important mechanism for the movement of contaminants and nutrients to remote locations, usually bird colonies, through excretion, molting and decomposition of carcasses. Methylmercury (MeHg) is a bioaccumulative neurotoxin and endocrine disruptor which is present in many remote ecosystems. We collected guano samples biweekly from a herring gull (Larus smithsoniansus) colony over two summers and analyzed MeHg, total mercury (THg), water-extractable phosphate (PO43-) and sulfate (SO42-). Concentrations of THg in guano declined through the summer months while %MeHg significantly increased (ranging from 12% to 100% of THg), suggesting a switch in diet as gull nutritional needs or food availability changed through the summer. The percentage of PO43- in dry guano increased throughout the summer (ranging from 2.8% to 4.4% of dry weight) and SO42- varied throughout the season (ranging from 0.1% to 0.8% of dry weight). These data indicate that gulls are transporting considerable amounts of MeHg, PO43-, and likely other contaminants to Big Meadow Bog, Nova Scotia.

Investigating Student Perceptions of Instructor Talk: Alignment with Researchers' Categorizations and Analysis of Remembered Language.

Instructor Talk-the noncontent language used by an instructor during class time-is likely to influence learning environments in science classrooms from the student perspective. Despite Instructor Talk being found in every science course thus far, investigations into student perceptions and memories of it are limited. We investigated to what extent undergraduate biology students 1) were aligned with researchers in their perceptions of Instructor Talk as Positively Phrased or Negatively Phrased and 2) remembered Instructor Talk. To test these ideas, we engaged 90 biology students in a multipart assessment. First, students were given randomly selected Instructor Talk quotes, half Positively Phrased and half Negatively Phrased, and were asked to evaluate each quote as promoting a positive or negative learning environment. Overall, students evaluated the Instructor Talk quotes similarly to researchers' categorizations (p < 0.0001). Second, students were asked to provide examples of remembered instructor language from their biology courses that they felt promoted a positive or negative learning environment. Most students shared multiple memories, and ∼75% of these memories could be coded with the Instructor Talk frameworks. Given that students perceive and remember Instructor Talk as impacting the learning environment, Instructor Talk may be an explanatory variable for differential student outcomes across studies of active learning.

Ruminal Protozoal Populations of Angus Steers Differing in Feed Efficiency.

Feed accounts for as much as 70% of beef production costs, and improvement of the efficiency with which animals convert feed to product has the potential to have substantial financial impact on the beef industry. The rumen microbiome plays a key role in determining feed efficiency; however, previous studies of rumen microbiota have not focused on protozoal communities despite the estimation that these organisms represent approximately 50% of rumen content biomass. Protozoal communities participate in the regulation of bacterial populations and nitrogen cycling-key aspects of microbiome dynamics. The present study focused on identifying potential associations of protozoal community profiles with feed efficiency. Weaned steers (n = 50) 7 months of age weighing approximately 260 kg were adapted to a growing ration and GrowSafe for 2 weeks prior to a 70-day feed efficiency trial. The GrowSafe system is a feeding system that monitors feed intake in real time. Body weights were collected on the first day and then every 7 days of the feed efficiency trial, and on the final day, approximately 50 mL of rumen content were collected via orogastric tubing and frozen at -80 °C. Body weight and feed intake were used to calculate residual feed intake (RFI) as a measure of feed efficiency, and steers were categorized as high (n = 14) or low (n = 10) RFI based on ±0.5 standard deviations about the mean RFI. Microbial DNA was extracted, and the eukaryotic component profiled by amplification and sequencing of 18S genes using degenerate primers that can amplify this locus across a range of protists. The taxonomy of protozoal sequences was assigned using QIIME 1.9 and analyzed using QIIME and SAS 9.4 with significance determined at α ≤ 0.05. Greater abundances of unassigned taxa were associated with high-RFI steers (p = 0.03), indicating a need for further study to identify component protozoal species. Differences were observed between low- and high-RFI steers in protozoal community phylogenetic diversity, including weighted beta-diversity (p = 0.04), Faith's phylogenetic diversity (p = 0.03), and observed Operational taxonomic unit (OTU) (p = 0.03). The unassigned taxa and differences in phylogenetic diversity of protozoal communities may contribute to divergences observed in feed efficiency phenotypes in beef steers.

Lessons learned from Hurricane Irma.

In the aftermath of Hurricane Irma, Emergency Managers looked at the challenges faced during the storm and found areas for improvement. Some of those challenges included evacuation procedures, staffing at the Emergency Operations Centers, slow debris removal times, and prolonged power outages. This research considers various counties impacted directly and indirectly by Hurricane Irma. The After-Action Reports of several counties have been summarized, including not only the problems, but recommendations for improving or preventing those problems from occurring again. The lessons learned from Hurricane Irma could lessen the impact of future storms, not only for those who dealt with Irma, but for all communities that can face high impact natural disasters.

Longitudinal Integration of Oral Health Training Into Physician Assistant Education.

PURPOSE: The purpose of this study was to evaluate physician assistant (PA) student knowledge after integrating an oral health curriculum longitudinally into a PA program and to assess student perceptions of the oral health curriculum. METHODS: This study was a 2-group longitudinal study comparing differences between the groups at 2 points in time. Cohort I (n = 60) took an oral health pretest and then, after completing oral health training, took a posttest. Cohort II (n = 49), which had not received oral health training, took identical testing on the same time interval. The students in Cohort I assessed the value of the oral health curriculum by answering a standardized perception survey. RESULTS: Cohort I had a statistically significant increase (p ≤ .0001) in posttest assessment scores compared to Cohort II (control group). The students who were exposed to the oral health curriculum reported a deeper understanding and appreciation of oral health disease and increased confidence in oral health clinical application. CONCLUSIONS: A greater awareness of the importance of oral health and chronic disease has evolved within PA education over the past 2 decades. Despite this progress, there is still a need to expand this curriculum and improve PA students' depth of knowledge in oral health. This study presents a novel and effective approach to incorporating an oral health curriculum into a PA program.