Pediatric Unintentional Fall-Related Injuries in a Statewide Trauma Registry.

OBJECTIVES: The purpose of the study was to evaluate patterns of fall-related injury through childhood and identify risk factors for more severe fall-related injuries with the goal of informing targeted prevention strategies for different ages. METHODS: The study population consisted of pediatric patients in the Iowa Trauma Registry from January 1, 2010, to December 31, 2014, who sustained an unintentional fall-related injury (N = 3856 patients). Multinomial logistic regression analysis was used to predict injury severity. Adjusted odds ratios were calculated characterizing the relationship between fall severity and age, sex, race, and fall type. RESULTS: More males (62%) sustained a fall-related injury during the study period when compared with females (38%; P < 0.0001). Head injuries were the most common type of injury in the younger than 1 year age group (77%), whereas fractures were the predominant injury type in all other age groups, followed by head injuries. Those younger than 1 year (adjusted odds ratio, 4.0; 95% confidence interval, 2.36-6.90) and aged 15 to 18 years (adjusted odds ratio, 1.9; 95% confidence interval, 1.17-3.03) were more likely to have an Injury Severity Score of ≥16 than those aged 10 to 14 years. CONCLUSIONS: Recommendations and prevention strategies need to focus on specific risk factors to reduce the harm of multilevel falls. As we have shown, patterns of fall injuries presenting to trauma hospitals differ by age, thus suggesting that prevention strategies focus on specific age groups.

Engineering a Single-Agent Cytokine/Antibody Fusion That Selectively Expands Regulatory T Cells for Autoimmune Disease Therapy.

IL-2 has been used to treat diseases ranging from cancer to autoimmune disorders, but its concurrent immunostimulatory and immunosuppressive effects hinder efficacy. IL-2 orchestrates immune cell function through activation of a high-affinity heterotrimeric receptor (composed of IL-2Rα, IL-2Rß, and common γ [γc]). IL-2Rα, which is highly expressed on regulatory T (TReg) cells, regulates IL-2 sensitivity. Previous studies have shown that complexation of IL-2 with the JES6-1 Ab preferentially biases cytokine activity toward TReg cells through a unique mechanism whereby IL-2 is exchanged from the Ab to IL-2Rα. However, clinical adoption of a mixed Ab/cytokine complex regimen is limited by stoichiometry and stability concerns. In this study, through structure-guided design, we engineered a single agent fusion of the IL-2 cytokine and JES6-1 Ab that, despite being covalently linked, preserves IL-2 exchange, selectively stimulating TReg expansion and exhibiting superior disease control to the mixed IL-2/JES6-1 complex in a mouse colitis model. These studies provide an engineering blueprint for resolving a major barrier to the implementation of functionally similar IL-2/Ab complexes for treatment of human disease.

Use of a Bidirectional Text Messaging System for Emergency Department Follow-Up Versus Usual Follow-Up.

Background: The use of text messaging is a growing trend. Usual care for follow-up with patients (no dedicated communication) has proven unreliable, and alternative communication methods may be beneficial. Introduction: The objective was to evaluate the effect of text messaging as a means of follow-up communication compared to usual care on patient satisfaction among patients discharged from the emergency department (ED). Materials and Methods: Participants completed a baseline survey about their text message usage and ED visit satisfaction. The participants completed a follow-up survey 2 weeks later. Participants randomized to text messaging received a text message at 24 h, 1 week, and 2 weeks after discharge. Control participants received usual care (typically no dedicated communication). Bivariate analyses were performed, and intent-to-treat and per protocol analyses were completed to examine follow-up satisfaction with ED communication/care. Results: A total of 802 subjects were recruited (text messaging-398 subjects, usual care-404 subjects). In the intent-to-treat analysis, text messaging subjects were not more likely to report satisfaction with follow-up communication (adjusted odds ratio [aOR] 0.90 [0.46-1.75]) or follow-up care (aOR 0.66 [0.30-1.46]) than usual care subjects. In per-protocol analysis, text messaging subjects had 2.95 (1.52-5.73) higher odds of reporting satisfaction with follow-up communication and 3.24 (1.46-7.16) higher odds of reporting satisfaction with follow-up care. Discussion: The use of text messaging for follow-up, when comparing satisfaction with follow-up communication and follow-up care after discharge, performs at least equally as well as usual follow-up. Conclusions: Text messaging is a provider time-saving and resource-conserving technology allowing health care providers to potentially reach a larger proportion of patients, making it a valuable form of follow-up communication.

Telemedicine Is Associated with Faster Diagnostic Imaging in Stroke Patients: A Cohort Study.

BACKGROUND: Meeting time goals for patients with time-sensitive conditions can be challenging in rural emergency departments (EDs), and adopting policies is critical. ED-based telemedicine has been proposed to improve quality and timeliness of care in rural EDs. INTRODUCTION: The objective of this study was to test the hypothesis that diagnostic testing in telemedicine-supplemented ED care for patients with myocardial infarction (MI) and stroke would be faster than nontelemedicine care in rural EDs. MATERIALS AND METHODS: This observational cohort study included all ED patients with MI or stroke in 19 rural critical access hospitals served by a single real-time contract-based telemedicine provider in the upper Midwest (2007-2015). The primary outcome for the MI cohort was time-to-electrocardiogram (EKG) and for the stroke cohort was time-to-head computed tomography (CT) interpretation. To measure the relationship between telemedicine and timeliness parameters, generalized estimating equations models were used, clustering on presenting hospital. RESULTS: Of participating ED visits, 756 were included in the MI cohort (29% used telemedicine) and 140 were included in the stroke cohort (30% used telemedicine). Time-to-EKG did not differ when telemedicine was used (1% faster, 95% confidence interval [CI] -4% to 7%), or after telemedicine was implemented (4% faster, 95% CI -3% to 10%). Head CT interpretation was faster for telemedicine cases (15% faster, 95% CI 4-26%). No differences were observed in time to reperfusion therapy. CONCLUSIONS: Telemedicine implementation was associated with more timely head CT interpretation for rural patients with stroke, but no difference in early MI care. Future work will focus on the specific manner in which telemedicine changes ED care processes and ongoing professional education.

Emergency Department Telemedicine Shortens Rural Time-To-Provider and Emergency Department Transfer Times.

BACKGROUND: Emergency department (ED)-based telemedicine has been implemented in many rural hospitals to provide specialty care and expertise to patients with critical time-sensitive conditions. INTRODUCTION: The purpose of this study was to measure the impact of ED-based telemedicine on timeliness of care in participating rural hospitals. MATERIALS AND METHODS: Matched cohort study of patients seen in 1 of 14 rural hospitals in a large Midwestern telemedicine network. Telemedicine cases were matched 2:1 with controls based on age, diagnosis, and hospital. The primary outcome was door-to-provider time, and secondary outcomes included ED length-of-stay (LOS) and time-to-transfer in those transferred to other hospitals. RESULTS: Of 127,928 qualifying ED encounters, 2,857 consulted telemedicine and were matched with nontelemedicine controls. Door-to-provider time was shorter in telemedicine patients by 6.0 min (95% confidence interval [CI] 4.3-7.8 min). The first provider seeing the patient was a telemedicine provider in 41.7% of telemedicine encounters, and in these cases, telemedicine was 14.7 min earlier than local providers. ED LOS was 22.1 min shorter (95% CI 3.1-41.2) among transferred patients, but total ED LOS was longer (40.2 min, 95% CI 30.8-49.6 min) for all telemedicine patients. CONCLUSIONS: Telemedicine decreases ED door-to-provider time, most commonly because the telemedicine provider was the first provider seeing a patient. Among transferred patients, ED LOS at the first hospital was shorter in patients who had telemedicine consulted. Future work will focus on the clinical impact of more timely rural ED care.

Steering teens safe: a randomized trial of a parent-based intervention to improve safe teen driving.

BACKGROUND: Crashes are the leading cause of death for teens, and parent-based interventions are a promising approach. We assess the effectiveness of Steering Teens Safe, a parent-focused program to increase safe teen driving. METHODS: Steering Teens Safe aimed to improve parental communication with teens about safe driving using motivational interviewing techniques in conjunction with 19 safe driving lessons. A randomized controlled trial involved 145 parent-teen dyads (70 intervention and 75 control). Intervention parents received a 45-minute session to learn the program with four follow-up phone sessions, a DVD, and a workbook. Control parents received a standard brochure about safe driving. Scores were developed to measure teen-reported quantity and quality of parental communication about safe driving. The main outcome measure was a previously validated Risky Driving Score reported by teens. Because the Score was highly skewed, a generalized linear model based on a gamma distribution was used for analysis. RESULTS: Intervention teens ranked their parent's success in talking about driving safety higher than control teens (p = 0.035) and reported that their parents talked about more topics (non-significant difference). The Risky Driving Score was 21% lower in intervention compared to control teens (85% CI = 0.60, 1.00). Interaction between communication quantity and the intervention was examined. Intervention teens who reported more successful communication had a 42% lower Risky Driving Score (95% CI = 0.37, 0.94) than control parents with less successful communication. CONCLUSIONS: This program had a positive although not strong effect, and it may hold the most promise in partnership with other programs, such as Driver's Education or Graduated Driver's License policies. TRIAL REGISTRATION: ClinicalTrials.gov NCT01014923. Registered Nov. 16, 2009.

Provider-to-provider telehealth for sepsis patients in a cohort of rural emergency departments.

BACKGROUND: Telehealth has been proposed as one strategy to improve the quality of time-sensitive sepsis care in rural emergency departments (EDs). The purpose of this study was to measure the association between telehealth-supplemented ED (tele-ED) care, health care costs, and clinical outcomes among patients with sepsis in rural EDs. METHODS: Cohort study using Medicare fee-for-service claims data for beneficiaries treated for sepsis in rural EDs between February 1, 2017, and September 30, 2019. Our primary hospital-level analysis used multivariable generalized estimating equations to measure the association between treatment in a tele-ED-capable hospital and 30-day total costs of care. In our supporting secondary analysis, we conducted a propensity-matched analysis of patients who used tele-ED with matched controls from non-tele-ED-capable hospitals. Our primary outcome was total health care payments among index hospitalized patients between the index ED visit and 30 days after hospital discharge, and our secondary outcomes included hospital mortality, hospital length of stay, 90-day mortality, 28-day hospital-free days, and 30-day inpatient readmissions. RESULTS: In our primary analysis, sepsis patients in tele-ED-capable hospitals had 6.7% higher (95% confidence interval [CI] 2.1%-11.5%) total health care costs compared to those in non-tele-ED-capable hospitals. In our propensity-matched patient-level analysis, total health care costs were 23% higher (95% CI 16.5%-30.4%) in tele-ED cases than matched non-tele-ED controls. Clinical outcomes were similar. CONCLUSIONS: Tele-ED capability in a mature rural tele-ED network was not associated with decreased health care costs or improved clinical outcomes. Future work is needed to reduce rural-urban sepsis care disparities and formalize systems of regionalized care.

Targeting Acute Myeloid Leukemia Stem Cells Through Perturbation of Mitochondrial Calcium.

Acute myeloid leukemia stem cells (LSCs) are uniquely reliant on oxidative phosphorylation (OXPHOS) for survival. Moreover, maintenance of OXPHOS is dependent on BCL-2, creating a therapeutic opportunity to target LSCs using the BCL-2 inhibitor venetoclax. While venetoclax-based regimens have shown promising clinical activity, the emergence of drug resistance is prevalent. Thus, in the present study, we investigated how mitochondrial properties may influence venetoclax responsiveness. Our data show that utilization of mitochondrial calcium is fundamentally different between drug-responsive and non-responsive LSCs. By comparison, venetoclax-resistant LSCs demonstrate a more active metabolic (i.e. OXPHOS) status with relatively high levels of calcium. Consequently, we tested genetic and pharmacological approaches to target the mitochondrial calcium uniporter, MCU. We demonstrate that inhibition of calcium uptake reduces OXPHOS and leads to eradication of venetoclax-resistant LSCs. These findings demonstrate a central role for calcium signaling in LSCs and provide an avenue for clinical management of venetoclax resistance.

Healthcare providers' knowledge, attitudes and counselling on injury prevention for preschool children in Croatia.

Injuries are the leading cause of death for children and young adults in Croatia. Research has indicated that health care providers can be effective in reducing the risk for traumatic injury through anticipatory guidance, but successful guidance requires that providers have injury knowledge and informed safety attitudes. This is the first study in Croatia to identify health care provider's knowledge, attitudes, and practices regarding anticipatory guidance on injury prevention for children. A stratified, random sample of licensed Croatian healthcare providers was mailed a survey, with a response of rate of 39.5 %. Participants included pediatricians, family physicians, gynecologists, each with a focus on primary care, and community nurses. Participants filled out a 15-minute paper-and-pencil survey that tested their knowledge of injury risks and prevention strategies, assessed their safety-prone attitudes, and measured the extent to which they counselled their patients on injury prevention. Pediatricians had the highest knowledge of injury risks and intervention approaches, with an average correct score of six out of ten (significantly higher than all other provider types). Knowledge was highest regarding infant fall risk and lowest for safe sleep positions. Pediatricians and community nurses had the highest safety-prone attitudes. Safety prone attitudes were strongest for transportation safety and weakest for safe sleeping position for all providers. Community nurses reported the highest level of patient counselling, followed by pediatricians. Both factual education and support in translating knowledge into everyday practice are necessary for health care providers. Implementing anticipatory guidance for child safety is a promising approach in Croatia.

Targeting Acute Myeloid Leukemia Stem Cells Through Perturbation of Mitochondrial Calcium.

We previously reported that acute myeloid leukemia stem cells (LSCs) are uniquely reliant on oxidative phosphorylation (OXPHOS) for survival. Moreover, maintenance of OXPHOS is dependent on BCL2, creating a therapeutic opportunity to target LSCs using the BCL2 inhibitor drug venetoclax. While venetoclax-based regimens have indeed shown promising clinical activity, the emergence of drug resistance is prevalent. Thus, in the present study, we investigated how mitochondrial properties may influence mechanisms that dictate venetoclax responsiveness. Our data show that utilization of mitochondrial calcium is fundamentally different between drug responsive and non-responsive LSCs. By comparison, venetoclax-resistant LSCs demonstrate a more active metabolic (i.e., OXPHOS) status with relatively high steady-state levels of calcium. Consequently, we tested genetic and pharmacological approaches to target the mitochondrial calcium uniporter, MCU. We demonstrate that inhibition of calcium uptake sharply reduces OXPHOS and leads to eradication of venetoclax-resistant LSCs. These findings demonstrate a central role for calcium signaling in the biology of LSCs and provide a therapeutic avenue for clinical management of venetoclax resistance.

A Novel Type of Monocytic Leukemia Stem Cell Revealed by the Clinical Use of Venetoclax-Based Therapy.

The BCL2 inhibitor venetoclax has recently emerged as an important component of acute myeloid leukemia (AML) therapy. Notably, use of this agent has revealed a previously unrecognized form of pathogenesis characterized by monocytic disease progression. We demonstrate that this form of disease arises from a fundamentally different type of leukemia stem cell (LSC), which we designate as monocytic LSC (m-LSC), that is developmentally and clinically distinct from the more well-described primitive LSC (p-LSC). The m-LSC is distinguished by a unique immunophenotype (CD34-, CD4+, CD11b-, CD14-, CD36-), unique transcriptional state, reliance on purine metabolism, and selective sensitivity to cladribine. Critically, in some instances, m-LSC and p-LSC subtypes can co-reside in the same patient with AML and simultaneously contribute to overall tumor biology. Thus, our findings demonstrate that LSC heterogeneity has direct clinical significance and highlight the need to distinguish and target m-LSCs as a means to improve clinical outcomes with venetoclax-based regimens. SIGNIFICANCE: These studies identify and characterize a new type of human acute myeloid LSC that is responsible for monocytic disease progression in patients with AML treated with venetoclax-based regimens. Our studies describe the phenotype, molecular properties, and drug sensitivities of this unique LSC subclass. This article is featured in Selected Articles from This Issue, p. 1949.

Analysis of acyclic nucleoside modifications in siRNAs finds sensitivity at position 1 that is restored by 5'-terminal phosphorylation both in vitro and in vivo.

Small interfering RNAs (siRNAs) are short, double-stranded RNAs that use the endogenous RNAi pathway to mediate gene silencing. Phosphorylation facilitates loading of a siRNA into the Ago2 complex and subsequent cleavage of the target mRNA. In this study, 2', 3' seco nucleoside modifications, which contain an acylic ribose ring and are commonly called unlocked nucleic acids (UNAs), were evaluated at all positions along the guide strand of a siRNA targeting apolipoprotein B (ApoB). UNA modifications at positions 1, 2 and 3 were detrimental to siRNA activity. UNAs at positions 1 and 2 prevented phosphorylation by Clp1 kinase, abrogated binding to Ago2, and impaired Ago2-mediated cleavage of the mRNA target. The addition of a 5'-terminal phosphate to siRNA containing a position 1 UNA restored ApoB mRNA silencing, Ago2 binding, and Ago2 mediated cleavage activity. Position 1 UNA modified siRNA containing a 5'-terminal phosphate exhibited a partial restoration of siRNA silencing activity in vivo. These data reveal the complexity of interpreting the effects of chemical modification on siRNA activity, and exemplify the importance of using multiple biochemical, cell-based and in vivo assays to rationally design chemically modified siRNA destined for therapeutic use.

Pediatric poisonings and risk markers for hospital admission in a major emergency department in Romania.

To identify the prevalence, characteristics, and risk markers for childhood poisonings treated in the emergency department of a large Romanian hospital. Trauma registry data using ICD-10 codes and case summaries were studied to identify poisonings among children aged 0-18. Multivariate logistic regression identified factors associated with hospital admission. Between 1999 and 2003, 1,173 pediatric trauma cases were seen in the emergency department; 437 (37.3%) were treated for poisoning, including medication (35%), alcohol (26%), chemical products (19%), and carbon monoxide (14%). Half of all poisonings were unintentional, primarily affecting children < 10 years. Half were intentional, mainly affecting children 10-18. Females were three times more likely than males to have documented suicidal intent (P < .0001). Over 30% of suicide attempts were among children ages 10-14 (P < .0001). We report significantly increased adjusted odds ratios (P < .05) of hospital admission for children 10-18, and for chemical substance poisoning, and suicidal intent. Pediatric poisoning is a serious public health issue in Romania, and we suggest these findings are relevant across other eastern European countries with limited resources. Poisonings result in morbidity and hospital admissions, yet there are few prevention resources available. Health education programs and consumer product safety policies are needed in Romania and eastern Europe.

Flood-related work disruption and poor health outcomes among university students.

INTRODUCTION: Globally, floods are the most common and among the most devastating of natural disasters. Natural disasters such as floods impact local businesses, increasing local unemployment by up to 8.2%. Previous research has linked individual losses from disasters with symptoms such as posttraumatic stress disorder. However, little is known about the impact of work disruption and job loss on post-disaster psychological symptoms. University students, who are often living far away from family support structures and have limited resources, may be particularly vulnerable. This study examines student psychological health following a large flood at a university. HYPOTHESIS: Students who experienced flood-related job loss or disruption had a higher proportion of psychological symptoms than those who did not experience job loss or disruption, controlling for individual loss such as injury, home loss or evacuation. METHODS: On June 8, 2008, a major flood affected seven US Midwestern states. A total of two dozen people were killed and 148 injured, although no deaths or serious injuries were reported in the population used for this study. At the study university, operations were closed for one week, and 20 buildings were severely damaged. A cross-sectional survey of all students enrolled during the semester of the flood was conducted. Students were sent an online survey six weeks after the flood. In addition to questions about damage to their homes, the survey asked students if their work was disrupted because of the floods. Symptoms of PTSD were measured through the modified Child PTSD Symptom Scale. RESULTS: Of the 1,231 responding students with complete surveys, 667 (54.2%) reported that their work was disrupted due to the floods. Controlling for gender, ethnicity, grade, and damage to the student's home, students reporting work disruption were more than four times more likely to report PTSD symptoms (95% CI, 2.5-8.2). Work disruption was independently associated with decreases in general mental and physical health following the floods, as well as with increases in alcohol use. CONCLUSION: Disaster research has focused on damage to individuals and homes, but there has been little focus on work losses. Individuals who lose their jobs may be a vulnerable population post-disaster.

An In Vitro Evaluation of Denture Cleansing Regimens against a Polymicrobial Denture Biofilm Model.

Denture stomatitis (DS) is an inflammatory disease resulting from a polymicrobial biofilm perturbation at the denture surface-palatal mucosa interface. Recommendations made by dental health care professionals often lack clarity for appropriate denture cleaning. This study investigated the efficacy of brushing with off-the-shelf denture cleanser (DC) tablets (Poligrip®) vs. two toothpastes (Colgate® and Crest®) in alleviating the viable microorganisms (bacteria and fungi) in an in vitro denture biofilm model. Biofilms were grown on poly(methyl)methacrylate (PMMA) discs, then treated daily for 7 days with mechanical disruption (brushing), plus Poligrip® DC, Colgate® or Crest® toothpastes. Weekly treatment with Poligrip® DC on day 7 only was compared to daily modalities. All treatment parameters were processed to determine viable colony forming units for bacteria and fungi using the Miles and Misra technique, and imaged by confocal laser scanning microscopy (CLSM). Brushing with daily DC therapy was the most effective treatment in reducing the viable biofilm over 7 days of treatment. Brushing only was ineffective in controlling the viable bioburden, which was confirmed by CLSM imaging. This data indicates that regular cleansing of PMMA with DC was best for polymicrobial biofilms.

South Yorkshire Cohort: a 'cohort trials facility' study of health and weight - protocol for the recruitment phase.

BACKGROUND: Growing levels of both obesity and chronic disease in the general population pose a major public health problem. In the UK, an innovative 'health and weight' cohort trials facility, the 'South Yorkshire Cohort', is being built in order to provide robust evidence to inform policy, commissioning and clinical decisions in this field. This protocol reports the design of the facility and outlines the recruitment phase methods. METHOD/DESIGN: The South Yorkshire Cohort health and weight study uses the cohort multiple randomised controlled trial design. This design recruits a large observational cohort of patients with the condition(s) of interest which then provides a facility for multiple randomised controlled trials (with large representative samples of participants, long term outcomes as standard, increased comparability between each trial conducted within the cohort and increased efficiency particularly for trials of expensive interventions) as well as ongoing information as to the natural history of the condition and treatment as usual.This study aims to recruit 20,000 participants to the population based South Yorkshire Cohort health and weight research trials facility. Participants are recruited by invitation letters from their General Practitioners. Data is collected using postal and/or online patient self completed Health Questionnaires. NHS numbers will be used to facilitate record linkage and access to routine data. Participants are eligible if they are: aged 16 - 85 years, registered with one of 40 practices in South Yorkshire, provide consent for further contact from the researchers and to have their information used to look at the benefit of health treatments. The first wave of data is being collected during 2010/12 and further waves are planned at 2 - 5 year intervals for the planned 20 year duration of the facility. DISCUSSION: The South Yorkshire Cohort combines the strengths of the standard observational, longitudinal cohort study design with a population based cohort facility for multiple randomised controlled trials in a range of long term health and weight related conditions (including obesity). This infrastructure will allow the rapid and cheap identification and recruitment of patients, and facilitate the provision of robust evidence to inform the management and self-management of health and weight.

Discovery of fragments inducing conformational effects in dynamic proteins using a second-harmonic generation biosensor.

Biophysical screening of compound libraries for the identification of ligands that interact with a protein is efficient, but does typically not reveal if (or how) ligands may interfere with its functional properties. For this a biochemical/functional assay is required. But for proteins whose function is dependent on a conformational change, such assays are typically complex or have low throughput. Here we have explored a high-throughput second-harmonic generation (SHG) biosensor to detect fragments that induce conformational changes upon binding to a protein in real time and identify dynamic regions. Multiwell plate format SHG assays were developed for wild-type and six engineered single-cysteine mutants of acetyl choline binding protein (AChBP), a homologue to ligand gated ion channels (LGICs). They were conjugated with second harmonic-active labels via amine or maleimide coupling. To validate the assay, it was confirmed that the conformational changes induced in AChBP by nicotinic acetyl choline receptor (nAChR) agonists and antagonists were qualitatively different. A 1056 fragment library was subsequently screened against all variants and conformational modulators of AChBP were successfully identified, with hit rates from 9-22%, depending on the AChBP variant. A subset of four hits was selected for orthogonal validation and structural analysis. A time-resolved grating-coupled interferometry-based biosensor assay confirmed the interaction to be a reversible 1-step 1 : 1 interaction, and provided estimates of affinities and interaction kinetic rate constants (K D = 0.28-63 µM, k a = 0.1-6 µM-1 s-1, k d = 1 s-1). X-ray crystallography of two of the fragments confirmed their binding at a previously described conformationally dynamic site, corresponding to the regulatory site of LGICs. These results reveal that SHG has the sensitivity to identify fragments that induce conformational changes in a protein. A selection of fragment hits with a response profile different to known LGIC regulators was characterized and confirmed to bind to dynamic regions of the protein.

Candida albicans as an Essential "Keystone" Component within Polymicrobial Oral Biofilm Models?

Background: Existing standardized biofilm assays focus on simple mono-species or bacterial-only models. Incorporating Candida albicans into complex biofilm models can offer a more appropriate and relevant polymicrobial biofilm for the development of oral health products. Aims: This study aimed to assess the importance of interkingdom interactions in polymicrobial oral biofilm systems with or without C. albicans, and test how these models respond to oral therapeutic challenges in vitro. Materials and Methods: Polymicrobial biofilms (two models containing 5 and 10 bacterial species, respectively) were created in parallel in the presence and absence of C. albicans and challenged using clinically relevant antimicrobials. The metabolic profiles and biomasses of these complex biofilms were estimated using resazurin dye and crystal violet stain, respectively. Quantitative PCR was utilized to assess compositional changes in microbial load. Additional assays, for measurements of pH and lactate, were included to monitor fluctuations in virulence "biomarkers." Results: An increased level of metabolic activity and biomass in the presence of C. albicans was observed. Bacterial load was increased by more than a factor of 10 in the presence of C. albicans. Assays showed inclusion of C. albicans impacted the biofilm virulence profiles. C. albicans did not affect the biofilms' responses to the short-term incubations with different treatments. Conclusions: The interkingdom biofilms described herein are structurally robust and exhibit all the hallmarks of a reproducible model. To our knowledge, these data are the first to test the hypothesis that yeasts may act as potential "keystone" components of oral biofilms.

Public health application of predictive modeling: an example from farm vehicle crashes.

BACKGROUND: The goal of predictive modelling is to identify the likelihood of future events, such as the predictive modelling used in climate science to forecast weather patterns and significant weather occurrences. In public health, increasingly sophisticated predictive models are used to predict health events in patients and to screen high risk individuals, such as for cardiovascular disease and breast cancer. Although causal modelling is frequently used in epidemiology to identify risk factors, predictive modelling provides highly useful information for individual risk prediction and for informing courses of treatment. Such predictive knowledge is often of great utility to physicians, counsellors, health education specialists, policymakers or other professionals, who may then advice course correction or interventions to prevent adverse health outcomes from occurring. In this manuscript, we use an example dataset that documents farm vehicle crashes and conventional statistical methods to forecast the risk of an injury or death in a farm vehicle crash for a specific individual or a scenario. RESULTS: Using data from 7094 farm crashes that occurred between 2005 and 2010 in nine mid-western states, we demonstrate and discuss predictive model fitting approaches, model validation techniques using external datasets, and the calculation and interpretation of predicted probabilities. We then developed two automated risk prediction tools using readily available software packages. We discuss best practices and common limitations associated with predictive models built from observational datasets. CONCLUSIONS: Predictive analysis offers tools that could aid the decision making of policymakers, physicians, and environmental health practitioners to improve public health.

Polymicrobial oral biofilm models: simplifying the complex.

Over the past century, numerous studies have used oral biofilm models to investigate growth kinetics, biofilm formation, structure and composition, antimicrobial susceptibility and host-pathogen interactions. In vivo animal models provide useful models of some oral diseases; however, these are expensive and carry vast ethical implications. Oral biofilms grown or maintained in vitro offer a useful platform for certain studies and have the advantages of being inexpensive to establish and easy to reproduce and manipulate. In addition, a wide range of variables can be monitored and adjusted to mimic the dynamic environmental changes at different sites in the oral cavity, such as pH, temperature, salivary and gingival crevicular fluid flow rates, or microbial composition. This review provides a detailed insight for early-career oral science researchers into how the biofilm models used in oral research have progressed and improved over the years, their advantages and disadvantages, and how such systems have contributed to our current understanding of oral disease pathogenesis and aetiology.