Logic-based mechanistic machine learning on high-content images reveals how drugs differentially regulate cardiac fibroblasts.

Fibroblasts are essential regulators of extracellular matrix deposition following cardiac injury. These cells exhibit highly plastic responses in phenotype during fibrosis in response to environmental stimuli. Here, we test whether and how candidate anti-fibrotic drugs differentially regulate measures of cardiac fibroblast phenotype, which may help identify treatments for cardiac fibrosis. We conducted a high-content microscopy screen of human cardiac fibroblasts treated with 13 clinically relevant drugs in the context of TGFß and/or IL-1ß, measuring phenotype across 137 single-cell features. We used the phenotypic data from our high-content imaging to train a logic-based mechanistic machine learning model (LogiMML) for fibroblast signaling. The model predicted how pirfenidone and Src inhibitor WH-4-023 reduce actin filament assembly and actin-myosin stress fiber formation, respectively. Validating the LogiMML model prediction that PI3K partially mediates the effects of Src inhibition, we found that PI3K inhibition reduces actin-myosin stress fiber formation and procollagen I production in human cardiac fibroblasts. In this study, we establish a modeling approach combining the strengths of logic-based network models and regularized regression models. We apply this approach to predict mechanisms that mediate the differential effects of drugs on fibroblasts, revealing Src inhibition acting via PI3K as a potential therapy for cardiac fibrosis.

Logic-based mechanistic machine learning on high-content images reveals how drugs differentially regulate cardiac fibroblasts.

Fibroblasts are essential regulators of extracellular matrix deposition following cardiac injury. These cells exhibit highly plastic responses in phenotype during fibrosis in response to environmental stimuli. Here, we test whether and how candidate anti-fibrotic drugs differentially regulate measures of cardiac fibroblast phenotype, which may help identify treatments for cardiac fibrosis. We conducted a high content microscopy screen of human cardiac fibroblasts treated with 13 clinically relevant drugs in the context of TGFß and/or IL-1ß, measuring phenotype across 137 single-cell features. We used the phenotypic data from our high content imaging to train a logic-based mechanistic machine learning model (LogiMML) for fibroblast signaling. The model predicted how pirfenidone and Src inhibitor WH-4-023 reduce actin filament assembly and actin-myosin stress fiber formation, respectively. Validating the LogiMML model prediction that PI3K partially mediates the effects of Src inhibition, we found that PI3K inhibition reduces actin-myosin stress fiber formation and procollagen I production in human cardiac fibroblasts. In this study, we establish a modeling approach combining the strengths of logic-based network models and regularized regression models, apply this approach to predict mechanisms that mediate the differential effects of drugs on fibroblasts, revealing Src inhibition acting via PI3K as a potential therapy for cardiac fibrosis.

Network model-based screen for FDA-approved drugs affecting cardiac fibrosis.

Cardiac fibrosis is a significant component of pathological heart remodeling, yet it is not directly targeted by existing drugs. Systems pharmacology approaches have the potential to provide mechanistic frameworks with which to predict and understand how drugs modulate biological systems. Here, we combine network modeling of the fibroblast signaling network with 36 unique drug-target interactions from DrugBank to predict drugs that modulate fibroblast phenotype and fibrosis. Galunisertib was predicted to decrease collagen and α-SMA expression, which we validated in human cardiac fibroblasts. In vivo fibrosis data from the literature validated predictions for 10 drugs. Further, the model was used to identify network mechanisms by which these drugs work. Arsenic trioxide was predicted to induce fibrosis by AP1-driven TGFß expression and MMP2-driven TGFß activation. Entresto (valsartan/sacubitril) was predicted to suppress fibrosis by valsartan suppression of ERK signaling and sacubitril enhancement of PKG activity, both of which decreased Smad3 activity. Overall, this study provides a framework for integrating drug-target mechanisms with logic-based network models, which can drive further studies both in cardiac fibrosis and other conditions.

Context-specific network modeling identifies new crosstalk in ß-adrenergic cardiac hypertrophy.

Cardiac hypertrophy is a context-dependent phenomenon wherein a myriad of biochemical and biomechanical factors regulate myocardial growth through a complex large-scale signaling network. Although numerous studies have investigated hypertrophic signaling pathways, less is known about hypertrophy signaling as a whole network and how this network acts in a context-dependent manner. Here, we developed a systematic approach, CLASSED (Context-specific Logic-bASed Signaling nEtwork Development), to revise a large-scale signaling model based on context-specific data and identify main reactions and new crosstalks regulating context-specific response. CLASSED involves four sequential stages with an automated validation module as a core which builds a logic-based ODE model from the interaction graph and outputs the model validation percent. The context-specific model is developed by estimation of default parameters, classified qualitative validation, hybrid Morris-Sobol global sensitivity analysis, and discovery of missing context-dependent crosstalks. Applying this pipeline to our prior-knowledge hypertrophy network with context-specific data revealed key signaling reactions which distinctly regulate cell response to isoproterenol, phenylephrine, angiotensin II and stretch. Furthermore, with CLASSED we developed a context-specific model of ß-adrenergic cardiac hypertrophy. The model predicted new crosstalks between calcium/calmodulin-dependent pathways and upstream signaling of Ras in the ISO-specific context. Experiments in cardiomyocytes validated the model's predictions on the role of CaMKII-Gßγ and CaN-Gßγ interactions in mediating hypertrophic signals in ISO-specific context and revealed a difference in the phosphorylation magnitude and translocation of ERK1/2 between cardiac myocytes and fibroblasts. CLASSED is a systematic approach for developing context-specific large-scale signaling networks, yielding insights into new-found crosstalks in ß-adrenergic cardiac hypertrophy.

Multiscale Coupling of an Agent-Based Model of Tissue Fibrosis and a Logic-Based Model of Intracellular Signaling.

Wound healing and fibrosis following myocardial infarction (MI) is a dynamic process involving many cell types, extracellular matrix (ECM), and inflammatory cues. As both incidence and survival rates for MI increase, management of post-MI recovery and associated complications are an increasingly important focus. Complexity of the wound healing process and the need for improved therapeutics necessitate a better understanding of the biochemical cues that drive fibrosis. To study the progression of cardiac fibrosis across spatial and temporal scales, we developed a novel hybrid multiscale model that couples a logic-based differential equation (LDE) model of the fibroblast intracellular signaling network with an agent-based model (ABM) of multi-cellular tissue remodeling. The ABM computes information about cytokine and growth factor levels in the environment including TGFß, TNFα, IL-1ß, and IL-6, which are passed as inputs to the LDE model. The LDE model then computes the network signaling state of individual cardiac fibroblasts within the ABM. Based on the current network state, fibroblasts make decisions regarding cytokine secretion and deposition and degradation of collagen. Simulated fibroblasts respond dynamically to rapidly changing extracellular environments and contribute to spatial heterogeneity in model predicted fibrosis, which is governed by many parameters including cell density, cell migration speeds, and cytokine levels. Verification tests confirmed that predictions of the coupled model and network model alone were consistent in response to constant cytokine inputs and furthermore, a subset of coupled model predictions were validated with in vitro experiments with human cardiac fibroblasts. This multiscale framework for cardiac fibrosis will allow for systematic screening of the effects of molecular perturbations in fibroblast signaling on tissue-scale extracellular matrix composition and organization.

Association of Mentor-to-Program Contact and Applicant Rank Disclosure With Vitreoretinal Fellowship Applicant's Final Match Outcome in 2016 and 2017.

Importance: Communication between applicants, mentors, and training programs is common before the residency and fellowship match. Few studies have examined the association of prematch communication on final match outcomes. Objectives: To report various characteristics of the vitreoretinal surgery fellowship match and to examine the association of mentor-to-program communication and applicant disclosure of their number 1 ranking with the probability of matching number 1. Design, Setting, and Participants: In this cross-sectional study of the 2016 and 2017 vitreoretinal surgery fellowship matches, an online survey examined (1) number of applications submitted, (2) number of programs ranked, (3) rank order of final match, (4) total application and interview-related costs, (5) mentor-to-program contact, (6) applicant disclosure of number 1 ranking, and (7) mentor disclosure of number 1 ranking. A link to an anonymous online survey was sent to 198 matched fellows (105 fellows from the 2016 match and 93 from the 2017 match). Main Outcomes and Measures: Survey responses regarding the vitreoretinal surgery fellowship application process. Results: The survey was sent to 198 matched fellowship applicants, and 152 (77%) completed the survey. Of the 105 matched applicants in 2016, 21 (20%) were women. Of the 93 matched applicants in 2017, 24 (26%) were women. Matched applicants applied to a mean (SD) of 34 (17) programs (range, 1-85) and ranked a mean (SD) of 12 (4) programs (range, 1-27). Of 152 applicants, 66 (43%) matched at their number 1 ranked program, 23 (15%) matched number 2, and 21 (14%) matched number 3. The mean (SD) total cost was $5500 ($2776) (range, $500-$13 500). Two-sided χ2 testing found no association (odds ratio, 0.70; 95% CI, 0.34-1.4; P = .33) between mentor-to-program contact and the probability of applicants matching at their number 1 ranked program. Matched applicants who revealed their number 1 ranking either personally or via a mentor matched at a program ranked lower (more desirable) on their rank list (mean match ranking, 2.8) compared with those who did not reveal their number 1 ranking (mean match ranking, 4.2; 95% CI, 0.2-2.5; P = .01). Applicant disclosure of their intention to rank a program number 1, either personally or via a mentor, was associated with matching number 1 (odds ratio, 2.6; range, 1.1-6.0; P = .03). Conclusions and Relevance: Vitreoretinal fellowship applicants who disclosed their number 1 ranking, either personally or via a mentor, were associated with improved match outcomes compared with their cohorts who did not make such disclosures.

Postintervention Effects of Click City® : Alcohol on Changing Etiological Mechanisms Related to the Onset of Heavy Drinking.

INTRODUCTION: Alcohol consumption, including heavy drinking, is the fourth leading preventable cause of death in the United States. Youth who engage in heavy drinking are likely to experience a number of problems associated with their use. In 2015, U.S. prevalence of heavy drinking was 17% among 12th graders. These data suggest a clear need for conducting alcohol prevention activities among youth. METHOD: We designed the Click City®: Alcohol program for 7th graders, with a booster in 8th grade. We conducted an efficacy trial in 25 schools in three counties in Oregon. Schools were randomized to either the Click City: Alcohol ( n = 12) or Usual Curriculum condition ( n = 13). We present the results of a short-term evaluation, assessing change in outcomes from baseline to 1 week following the intervention among students in Click City: Alcohol schools versus those in Usual Curriculum schools. RESULTS: Students who used the Click City: Alcohol program significantly decreased their intentions to drink heavily in the future, as compared with students in the Usual Curriculum control condition, although the effect size was small. Changes in the targeted mechanisms were in the expected direction and were significant for all but one mechanism, with moderate effect sizes. CONCLUSION: Our short-term findings provide preliminary support for the efficacy of the Click City: Alcohol program to change adolescents' intentions to engage in heavy drinking. If the results are maintained over time, the program has the potential to prevent the onset of heavy drinking among teens and reduce the negative consequences associated with heavy drinking, including neurological and other health consequences.

Lessons learned in the development and evaluation of RxCoach™, an mHealth app to increase tobacco cessation medication adherence.

OBJECTIVE: In this project we developed and evaluated a mobile health app to improve adherence to tobacco cessation medication. METHODS: The study was conducted in three phases: (1) Create app with input from our consultant, focus groups and user testing; (2) Test feasibility of the app; and (3) Develop and user-test the barcode scanner. RESULTS: Focus group feedback was instrumental in developing content and creating the user interface. User testing helped to identify problems and refine the app. The feasibility trial provided "real world" testing. We experienced challenges in recruitment due to the inclusion criteria. We had high attrition due to technical issues, medication side effects, enrollment procedures, and lack of personal contact. Among the five retained participants, use of the app was associated with good medication adherence and high consumer satisfaction. CONCLUSION: The small sample size limits the generalizability of the findings and the conclusions that can be drawn from the study. However, the feasibility trial enabled the team to identify ways to improve the conduct of this and other mHealth studies. PRACTICAL IMPLICATIONS: We should expand RxCoach to include all prescription and over-the-counter tobacco cessation medications, and re-test for feasibility using lessons learned to improve recruitment and retention.

Home medication support for childhood cancer: family-centered design and testing.

PURPOSE: Errors in the use of medications at home by children with cancer are common, and interventions to support correct use are needed. We sought to (1) engage stakeholders in the design and development of an intervention to prevent errors in home medication use, and (2) evaluate the acceptability and usefulness of the intervention. METHODS: We convened a multidisciplinary team of parents, clinicians, technology experts, and researchers to develop an intervention using a two-step user-centered design process. First, parents and oncologists provided input on the design. Second, a parent panel and two oncology nurses refined draft materials. In a feasibility study, we used questionnaires to assess usefulness and acceptability. Medication error rates were assessed via monthly telephone interviews with parents. RESULTS: We successfully partnered with parents, clinicians, and IT experts to develop Home Medication Support (HoMeS), a family-centered Web-based intervention. HoMeS includes a medication calendar with decision support, a communication tool, adverse effect information, a metric conversion chart, and other information. The 15 families in the feasibility study gave HoMeS high ratings for acceptability and usefulness. Half recorded information on the calendar to indicate to other caregivers that doses were given; 34% brought it to the clinic to communicate with their clinician about home medication use. There was no change in the rate of medication errors in this feasibility study. CONCLUSION: We created and tested a stakeholder-designed, Web-based intervention to support home chemotherapy use, which parents rated highly. This tool may prevent serious medication errors in a larger study.

Current options in the management of pellucid marginal degeneration.

PURPOSE: To review and evaluate current and future directions in the diagnosis and surgical management of pellucid marginal degeneration (PMD), including penetrating keratoplasty, full-thickness crescentic wedge resection (FTCWR), deep anterior lamellar keratoplasty (DALK), crescentic lamellar wedge resection (CLWR), crescentic lamellar keratoplasty, tuck-in lamellar keratoplasty (TILK), toric phakic intraocular lens (PIOL) implantation, intrastromal corneal ring segment implantation (ICRS), corneal collagen cross-linking (CXL), and combined therapies. This is the first review article looking at the literature specific to PMD. METHODS: Review of published studies. RESULTS: Reported data for each treatment is presented. Penetrating keratoplasty is the treatment of last resort in PMD and is effective, but with considerable complications. DALK provides visual outcomes similar to penetrating keratoplasty without the risk of immune-mediated graft rejection, but its complexity and relative novelty limit its acceptance. FTCWR has good visual outcomes, but with significant astigmatic drift. CLWR is effective, but lacks long-term results. Crescentic lamellar keratoplasty and TILK are effective, but technically difficult and without long-term results. Toric PIOL implantation is effective, but ectasia progression is a concern. ICRS implantation can delay penetrating keratoplasty and improve contact lens tolerance, but does not treat the underlying process. CXL demonstrates effectiveness without complications, although data are limited and long-term results are needed. Combining treatments such as ICRS, CXL, toric PIOL implantation, and refractive surgery is promising, but additional studies are needed to investigate their efficacy and safety. CONCLUSIONS: Although little is understood about the etiology, pathophysiology, epidemiology, and genetics of PMD, new treatments are improving visual outcomes and reducing complications. Corneal collagen cross-linking is especially exciting because it halts disease progression. Combined treatments and improved screening could eliminate the need for surgical management in most cases of PMD.

Change in keratometry after myopic laser in situ keratomileusis and photorefractive keratectomy.

PURPOSE: To compare the change in keratometry (K), spherical equivalent (SE), and visual acuity after myopic laser in situ keratomileusis (LASIK) and photorefractive keratectomy (PRK). SETTING: Academic tertiary care center. DESIGN: Retrospective review. METHODS: The postoperative K, SE, and uncorrected and corrected distance visual acuities were measured 6 months, 9 months, 1 year, 2 years, 3 years, 4 to 5 years, 6 to 7 years, and 8+ years postoperatively. A difference (Δ) for each variable was calculated from its 6-month postoperative baseline. The rates of change were grouped based on the magnitude of myopic correction (0.00 to 2.99 diopters [D]; 3.00 to 5.99 D; 6.00 to 8.99 D), type of surgery (LASIK versus PRK), and age (<34 years; 34 to 45 years; >45 years). RESULTS: Statistically significant differences were found in the rates of change between low and moderate corrections to high corrections for ΔKavg (P=.0472 and P=.0091, respectively) and ΔSE (both P<.0001). Statistically significant differences were found in the rate of change in ΔKavg between all 3 ages groups (P=.0330, P=.0051, and P<.0001) and in ΔSE between ages less than 34 years and 34 to 45 years to ages over 45 years (P=.0158 and P=.0015, respectively). There was no significant difference in the rate of change in ΔKavg and ΔSE between LASIK and PRK (P=.3599 and P=.9403, respectively). CONCLUSION: There was keratometric and refractive regression for myopic LASIK, with the rate of regression depending on treatment magnitude and age. FINANCIAL DISCLOSURE: No author has a financial or proprietary interest in any material or method mentioned.

Long-term efficacy of click city(r): tobacco: a school-based tobacco prevention program.

INTRODUCTION: Click City (®) : Tobacco is an innovative, computer-based tobacco prevention program designed to be implemented in 5th-grade classrooms with a booster in 6th grade. The program targets etiological mechanisms predictive of future willingness and intentions to use tobacco and initiation of tobacco use. Each component was empirically evaluated to assure that it changed its targeted mechanism. This paper describes long-term outcomes for students who participated in a randomized controlled efficacy trial of the program. METHODS: A total of 26 middle schools were stratified and randomly assigned to the Click City (®) : Tobacco program or Usual Curriculum. The 47 elementary schools that fed into each middle school were assigned to the same condition as their respective middle school. In Click City (®) : Tobacco schools, 1,168 students from 24 elementary schools and 13 middle schools participated. In Usual Curriculum schools, 1,154 students from 23 elementary schools and 13 middle schools participated. All participating students completed baseline, post-6th grade program, and 7th grade assessments. RESULTS: As compared to students in schools that continued with their usual curriculum, intentions and willingness to smoke increased less from baseline to 6th grade and from baseline to 7th grade, among students in schools that used the Click City (®) : Tobacco curriculum. Changes in mechanisms were also in the expected direction. The program was particularly efficacious for at-risk students. CONCLUSIONS: Results provide evidence to support the long-term efficacy of Click City (®) : Tobacco. Program development, based on an empirical evaluation of each component, most likely played a role in the success of the program.

Vitreous and Aqueous Penetration of Orally Administered Trimethoprim-Sulfamethoxazole Combination in Humans.

PURPOSE: To determine the penetration of orally administered trimethoprim (TMP)-sulfamethoxazole (SMX) into the aqueous and vitreous cavity of noninflamed human eyes. METHODS: Nine adult patients undergoing cataract surgery and 10 adult patients undergoing pars plana vitrectomy were given 3 doses of oral TMP-SMX every 12 hours before the surgery. Aqueous and blood samples were collected from patients undergoing cataract surgery; vitreous and blood samples were collected from patients undergoing vitrectomy. The levels of TMP and SMX were analyzed using high-performance liquid chromatography and were compared with the mean minimum inhibitory concentrations (MIC) of potential ocular pathogens. RESULTS: TMP-SMX was present in all samples. Among eyes undergoing cataract surgery, the mean concentrations of TMP in aqueous and blood were 0.341 ± 0.141 µg/mL (mean ± SD) and 1.501 ± 0.433 µg/mL and of SMX were 5.259 ± 0.929 µg/mL and 11.835 ± 2.100 µg/mL, respectively. Among eyes undergoing vitrectomy, the mean concentrations of TMP in vitreous and blood were 1.864 ± 0.807 µg/mL and 4.591 ± 2.979 µg/mL and of SMX were 5.910 ± 2.705 µg/mL and 39.289 ± 15.469 µg/mL, respectively. MIC levels were achieved against many bacterial pathogens, including methicillin-resistant Staphylococcus aureus. CONCLUSIONS: TMP-SMX penetrates both the aqueous and vitreous cavities when given orally. The components reach therapeutic inhibitory concentrations in the ocular cavity against many potential pathogens.

Corneal biomechanics in iatrogenic ectasia and keratoconus: A review of the literature.

The Ocular Response Analyzer (ORA) (Reichert Ophthalmic Instruments, Buffalo, NY) allows direct measurement of corneal biomechanical properties. Since its introduction, many studies have sought to elucidate the clinical applications of corneal hysteresis (CH) and corneal resistance factor (CRF). More recently, detailed corneal deformation signal waveform analysis (WA) has potentially expanded the diagnostic capabilities of the ORA. In this review, the role of CH, CRF, and WA are examined in keratoconus (KC) and iatrogenic ectasia (IE). The PubMed database was searched electronically for peer-reviewed literature in July 2012 and August 2012 without date restrictions. The search strategy included medical subject heading (MeSH) and natural language terms to retrieve references on corneal biomechanics, CH, CRF, corneal deformation signal WA, IE, and KC. The evidence suggests that while CH and CRF are poor screening tools when used alone, increased sensitivity and specificity of KC and IE screening result when these parameters are combined with tomography and topography. Recent advances in WA are promising, but little is currently understood about its biomechanical and clinical relevance. Future studies should seek to refine the screening protocols for KC and IE as well as define the clinical applicability of WA parameters.

Sources of medical error in refractive surgery.

PURPOSE: To evaluate the causes of laser programming errors in refractive surgery and outcomes in these cases. METHODS: In this multicenter, retrospective chart review, 22 eyes of 18 patients who had incorrect data entered into the refractive laser computer system at the time of treatment were evaluated. Cases were analyzed to uncover the etiology of these errors, patient follow-up treatments, and final outcomes. The results were used to identify potential methods to avoid similar errors in the future. RESULTS: Every patient experienced compromised uncorrected visual acuity requiring additional intervention, and 7 of 22 eyes (32%) lost corrected distance visual acuity (CDVA) of at least one line. Sixteen patients were suitable candidates for additional surgical correction to address these residual visual symptoms and six were not. Thirteen of 22 eyes (59%) received surgical follow-up treatment; nine eyes were treated with contact lenses. After follow-up treatment, six patients (27%) still had a loss of one line or more of CDVA. Three significant sources of error were identified: errors of cylinder conversion, data entry, and patient identification error. CONCLUSION: Twenty-seven percent of eyes with laser programming errors ultimately lost one or more lines of CDVA. Patients who underwent surgical revision had better outcomes than those who did not. Many of the mistakes identified were likely avoidable had preventive measures been taken, such as strict adherence to patient verification protocol or rigorous rechecking of treatment parameters.

Web-based family intervention for overweight children: a pilot study.

BACKGROUND: Research has shown the efficacy of family-based behavioral interventions for overweight children, but a major difficulty is access to effective treatment programs. The objective of this study was to develop and test the initial feasibility and efficacy of a web-based family program for overweight 8- to 12-year-old children. METHODS: A website was created using concepts from effective family-based behavioral programs and input from focus groups with overweight children, parents, and pediatricians. The website provided information about obesity and healthy lifestyles, assessment of dietary and physical activity habits, interactive dietary and physical activity games, and instruction in goal-setting and monitoring of goals. Children selected a dietary and physical activity goal and a daily step goal with pedometers. Feasibility and pilot testing over 4 weeks was conducted with 24 overweight children referred by a physician. Outcomes were z-BMI, healthy eating and physical activity, and intrinsic motivation and self-efficacy for weight control. RESULTS: Mean number of logins over the study period was 11.4 for the study sample. Eighteen families (75%) returned for the follow-up assessment. Pre-post analyses for these participants showed improvements in intrinsic motivation, (p=0.05), self-efficacy (p=0.025), physical activity (p=0.005), and healthy lifestyle behaviors (p=0.001). Comparisons between high and low users of the program indicated that high users reduced their BMI while low users increased their BMI over time (p=0.02); high users also improved their dietary intake relative to low users (p=0.04). Consumer satisfaction ratings were high. CONCLUSION: These pilot findings suggest this is a feasible approach for treatment of overweight children and that children who used the web program frequently improved their BMI and dietary intake.

A randomized clinical trial of a web-based tobacco cessation education program.

OBJECTIVES: We report the results of a randomized clinical trial of a 3-hour, web-based, tobacco cessation education program, the Web-Based Respiratory Education About Tobacco and Health (WeBREATHe) program, for practicing pediatric respiratory therapists (RTs), registered nurses (RNs), and nurse practitioners (NPs). METHODS: Two hundred fifteen RTs (n = 40), RNs (n = 163), and NPs (n = 12) employed at the Children's Hospital of Philadelphia and the Children's Hospital, University of Colorado at Denver, participated in this study. All study activities were completed online. After consenting, participants were randomly assigned to either the training (intervention) or delayed training (control) condition. The training condition consisted of a 3-hour continuing education unit course plus ongoing online resources. Participants were assessed at baseline, 1 week, and 3 months after enrollment. RESULTS: Participants in the training condition were more likely to increase their tobacco cessation intervention behaviors than their delayed training counterparts (F[1, 213] = 32.03, P < .001). Training participants showed significantly greater levels of advise (F[1, 213] = 7.22, P < .001); assess (F[1, 213] = 19.56, P < .001); and particularly assist/arrange (F[1213] = 35.52, P < .001). In addition, training condition participants rated the program highly on measures of consumer satisfaction. CONCLUSIONS: The WeBREATHe program is the first evidence-based education program in tobacco cessation designed specifically for pediatric RTs, RNs, and NPs. Engagement in WeBREATHe increased participants' tobacco cessation-related behaviors.