Development of PainFace software to simplify, standardize, and scale up mouse grimace analyses.

ABSTRACT: Facial grimacing is used to quantify spontaneous pain in mice and other mammals, but scoring relies on humans with different levels of proficiency. Here, we developed a cloud-based software platform called PainFace (http://painface.net) that uses machine learning to detect 4 facial action units of the mouse grimace scale (orbitals, nose, ears, whiskers) and score facial grimaces of black-coated C57BL/6 male and female mice on a 0 to 8 scale. Platform accuracy was validated in 2 different laboratories, with 3 conditions that evoke grimacing-laparotomy surgery, bilateral hindpaw injection of carrageenan, and intraplantar injection of formalin. PainFace can generate up to 1 grimace score per second from a standard 30 frames/s video, making it possible to quantify facial grimacing over time, and operates at a speed that scales with computing power. By analyzing the frequency distribution of grimace scores, we found that mice spent 7x more time in a "high grimace" state following laparotomy surgery relative to sham surgery controls. Our study shows that PainFace reproducibly quantifies facial grimaces indicative of nonevoked spontaneous pain and enables laboratories to standardize and scale-up facial grimace analyses.

Neuroimaging findings in 4342 hospitalized COVID-19 subjects: A multicenter report from the United States and Europe.

BACKGROUND AND PURPOSE: To determine the incidence of acute neuroimaging (NI) findings and comorbidities in the coronavirus disease of 2019 (COVID-19)-infected subjects in seven U.S. and four European hospitals. METHODS: This is a retrospective study of COVID-19-positive subjects with the following inclusion criteria: age >18, lab-confirmed COVID-19 infection, and acute NI findings (NI+) attributed to COVID-19 on CT or MRI brain. NI+ and comorbidities in total hospitalized COVID-19-positive (TN) subjects were assessed. RESULTS: A total of 37,950 COVID-19-positive subjects were reviewed and 4342 underwent NI. NI+ incidence in subjects with NI was 10.1% (442/4342) including 7.9% (294/3701) in the United States and 22.8% (148/647) in Europe. NI+ incidence in TN was 1.16% (442/37,950). In NI (4342), incidence of ischemic stroke was 6.4% followed by intracranial hemorrhage (ICH) (3.8%), encephalitis (0.5%), sinus venous thrombosis (0.2%), and acute disseminated encephalomyelitis (ADEM) (0.2%). White matter involvement was seen in 57% of NI+. Hypertension was the most common comorbidity (54%) before cardiac disease (28.8%) and diabetes mellitus (27.7%). Cardiac disease (p < .025), diabetes (p < .014), and chronic kidney disease (p < .012) were more common in the United States. CONCLUSION: This multicenter, multinational study investigated the incidence and spectrum of NI+ in 37,950 hospitalized adult COVID-19 subjects including regional differences in incidences of NI+, associated comorbidities, and other demographics. NI+ incidence in TN was 1.16% including 0.95% in the United States and 2.09% in Europe. ICH, encephalitis, and ADEM were common in Europe, while ischemic strokes were more common in the United States. In this cohort, incidence and distribution of NI+ helped characterize the neurological complications of COVID-19.

Consensus on a netball video analysis framework of descriptors and definitions by the netball video analysis consensus group.

Using an expert consensus-based approach, a netball video analysis consensus (NVAC) group of researchers and practitioners was formed to develop a video analysis framework of descriptors and definitions of physical, technical and contextual aspects for netball research. The framework aims to improve the consistency of language used within netball investigations. It also aims to guide injury mechanism reporting and identification of injury risk factors. The development of the framework involved a systematic review of the literature and a Delphi process. In conjunction with commercially used descriptors and definitions, 19 studies were used to create the initial framework of key descriptors and definitions in netball. In a two round Delphi method consensus, each expert rated their level of agreement with each of the descriptors and associated definition on a 5-point Likert scale (1-strongly disagree; 2-somewhat disagree; 3-neither agree nor disagree; 4-somewhat agree; 5-strongly agree). The median (IQR) rating of agreement was 5.0 (0.0), 5.0 (0.0) and 5.0 (0.0) for physical, technical and contextual aspects, respectively. The NVAC group recommends usage of the framework when conducting video analysis research in netball. The use of descriptors and definitions will be determined by the nature of the work and can be combined to incorporate further movements and actions used in netball. The framework can be linked with additional data, such as injury surveillance and microtechnology data.

Hospital transfer rates and advance care planning following a nursing home-targeted video-conference education series (Project ECHO): a prospective cohort study.

AIM: What effect does a novel education programme have on emergency hospital transfers of, and advance care planning decisions among, nursing home residents? FINDINGS: This education programme did not affect overall rates of emergency hospital transfer. It did increase advance care planning discussions, increase compliance with the results of these discussions and increase "DNR" orders among nursing home residents. MESSAGE: Novel tele-education programmes have the potential to improve advance care planning discussions in nursing homes.

Detection of after-discharges during intraoperative functional brain mapping in awake brain tumor surgery using a novel high-density circular grid.

OBJECTIVE: To evaluate intraoperative use of a novel high-density circular grid in detecting after-discharges (AD) on electrocorticography (ECoG) during functional brain mapping (FBM). METHODS: FBM during glioma surgery (10/2016 to 5/2019) recorded ADs using a 22-channel circular grid compared to conventional strip electrodes. ADs were analyzed for detection, duration, amplitude, morphology, histology, direction, and clinical signs. RESULTS: Thirty-two patients (mean age 54.2 years; r = 30-75) with glioma (WHO grade II-IV; 20 grade IV) had surgery. ADs during FBM were more likely in patients with wild-type as opposed to IDH-1 mutants (p < 0.0001) using more contacts compared with linear strip electrodes (p = 0.0001). More sensors tended to be involved in ADs detected by the circular grid vs strips (6.61 vs 3.43; p = 0.16) at lower stimulus intensity (3.14 mA vs 4.13 mA; p = 0.09). No difference in the number of cortical stimulations before resection was present (38.9 mA vs 47.9 mA; p = 0.26). ADs longer than 10 seconds were 32.5 seconds (circular grid) vs 58.4 (strips) (p = 0.12). CONCLUSIONS: High-density circular grids detect ADs in 360 degrees during FBM for glioma resection. Provocation of ADs was more likely in patients with wild-type than IDH-1 mutation. SIGNIFICANCE: Circular grids offer high-resolution ECoG during intraoperative FBM for detection of ADs.

Vulnerability to fatal drowning among the population in Southern Bangladesh: findings from a cross-sectional household survey.

OBJECTIVES: This study aimed to determine the fatal drowning burden and associated risk factors in Southern Bangladesh. SETTINGS: The survey was conducted in 39 subdistricts of all 6 districts of the Barisal division, Southern Bangladesh. PARTICIPANTS: All residents (for a minimum 6 months prior to survey) of the Barisal division, Southern Bangladesh. INTERVENTION/METHODS: A cross-sectional, divisionally representative household survey was conducted in all six districts of the Barisal division between September 2016 and February 2017, covering a population of 386 016. Data were collected by face-to-face interview with adult respondents using handheld electronic tablets. International Classification of Diseases (ICD)-v. 10 (ICD-10) Chapter XX: External causes of morbidity and mortality codes for drowning, W65-W74, X36-X39, V90, V92, X71 or X92, were used as the operational definition of a drowning event. RESULTS: The overall fatal drowning rate in Barisal was 37.9/100 000 population per year (95% CI 31.8 to 43.9). The highest fatal drowning rate was observed among children aged 1-4 years (262.2/100 000/year). Mortality rates among males (48.2/100 000/year) exceeded that for females (27.9/100 000/year). A higher rate of fatal drowning was found in rural (38.9/100 000/year) compared with urban areas (29.3/100 000/year). The results of the multivariable logistic regression identified that the factors significantly associated with fatal drowning were being male (OR 1.7, 95% CI 1.2 to 2.3), aged 1-4 years (OR 3.0, 95% CI 1.4 to 6.4) and residing in a household with four or more children (four or more children OR 1.8, 95% CI 1.1 to 2.9; and five or more children OR 2.1, 95% CI 1.2 to 3.7). CONCLUSION: Drowning is a public health problem, especially for children, in the Barisal division of Southern Bangladesh. Male gender, children 1-4 years of age and residing in a household with four or more children were associated with increased risk of fatal drowning events. The Barisal division demands urgent interventions targeted at high-risk groups identified in the survey.

Estimating drowning mortality in Tanzania: a systematic review and meta-analysis of existing data sources.

BACKGROUND: The WHO advocates a 7-step process to enable countries to develop and implement drowning prevention strategies. We sought to assess, using existing data sources, the drowning situation in Tanzania as a first step in this process. METHODS: We searched for data on causes of death in Tanzania by reviewing existing literature and global datasets and by in-country networking. Authors and institutions were then contacted to request aggregate data on drowning mortality. Site-specific drowning estimates were combined using a random effects meta-analytic approach. We also tested for evidence of variations in drowning estimates by sex and by age group. RESULTS: We acquired partial or complete information on drowning deaths for 13 data sources. We found strong evidence for substantial variations between study sites (p<0.001). Combining population-based data, we estimated an average of 5.1 drowning deaths per 100 000 persons per year (95% CI 3.8 to 6.3). The proportions of deaths due to drowning were 0.72% (95% CI 0.55 to 0.88) and 0.94% (95% CI 0.09 to 1.78) combining population-based data and hospital-based data, respectively. Males were at greater risk than females, while both under-five children and adults aged 45 years or more were at greater risk than those aged 5-44 years. CONCLUSION: Our estimates of drowning burden are broadly in line with the 2016 Global Burden of Disease and the 2015 WHO Global Health Estimates. While this exercise was useful in raising the burden of drowning in Tanzania with policy makers, planning drowning prevention strategies in this country will require a better understanding of which subpopulations are at high risk.

The IONA® Test: Development of an Automated Cell-Free DNA-Based Screening Test for Fetal Trisomies 13, 18, and 21 That Employs the Ion Proton Semiconductor Sequencing Platform.

OBJECTIVE: To develop a screening test for fetal trisomy 13, 18, and 21 using cell-free DNA from maternal blood with an automated workflow using the Ion Proton sequencing platform. METHODS: An automated next-generation sequencing workflow was developed using the Ion Proton sequencing platform and software developed for straightforward bioinformatic analysis. An algorithm was developed using 239 samples to determine the likelihood of trisomy, using DNA fragment counts and a fetal fraction validity check; the results were compared with those from invasive diagnostic procedures. A further 111 samples were used to assess the tests' sensitivity (detection rate) and specificity (1 minus false-positive rate). RESULTS: The 110 of a possible 111 valid samples used to verify the IONA® test gave 100% sensitivity and specificity, compared with invasive diagnostic procedures; one failed the fetal fraction validity check giving a sample failure rate of 0.29% across all 350 analysed samples. CONCLUSION: The data indicate that the IONA test provides a robust, accurate automated workflow suitable for use on maternal blood samples to screen for trisomies 13, 18, and 21. The test has the potential to reduce the number of unnecessary invasive procedures performed and facilitate testing by screening laboratories.

Data in support of a central role of plasminogen activator inhibitor-2 polymorphism in recurrent cardiovascular disease risk in the setting of high HDL cholesterol and C-reactive protein using Bayesian network modeling.

Data is presented that was utilized as the basis for Bayesian network modeling of influence pathways focusing on the central role of a polymorphism of plasminogen activator inhibitor-2 (PAI-2) on recurrent cardiovascular disease risk in patients with high levels of HDL cholesterol and C-reactive protein (CRP) as a marker of inflammation, "Influences on Plasminogen Activator Inhibitor-2 Polymorphism-Associated Recurrent Cardiovascular Disease Risk in Patients with High HDL Cholesterol and Inflammation" (Corsetti et al., 2016; [1]). The data consist of occurrence of recurrent coronary events in 166 post myocardial infarction patients along with 1. clinical data on gender, race, age, and body mass index; 2. blood level data on 17 biomarkers; and 3. genotype data on 53 presumptive CVD-related single nucleotide polymorphisms. Additionally, a flow diagram of the Bayesian modeling procedure is presented along with Bayesian network subgraphs (root nodes to outcome events) utilized as the data from which PAI-2 associated influence pathways were derived (Corsetti et al., 2016; [1]).

Influences on plasminogen activator inhibitor-2 polymorphism-associated recurrent cardiovascular disease risk in patients with high HDL cholesterol and inflammation.

BACKGROUND AND AIMS: Evidence continues to accumulate that athero-protective effects of high-density lipoprotein (HDL) depend to some degree on effective HDL functionality and that such functionality can become degraded in the setting of chronic inflammation. To investigate this issue, we have studied a group of post-myocardial infarction patients with high levels of C-reactive protein as an indicator of chronic inflammation and with concurrently high levels of HDL cholesterol. For these patients we have demonstrated high-risk for recurrent cardiac events as well as a strong association of risk with a polymorphism of the gene (SERPINB2) for plasminogen activator inhibitor-2 (PAI-2) presumptively reflective of an important role for fibrinolysis in risk. However, additional processes might be involved. The current work sought to characterize processes underlying how PAI-2 might be involved in the generation of risk. METHODS: Multivariate population data were leveraged using Bayesian network modeling, a graphical probabilistic approach for knowledge discovery, to generate networks reflective of influences on PAI-2 polymorphism-associated risk. RESULTS: Modeling results revealed three individual networks centering on the PAI-2 polymorphism with specific features providing information relating to how the polymorphism might associate with risk. These included racial dependency, platelet clot initiation and propagation, oxidative stress, inflammation effects on HDL metabolism and coagulation, and induction and termination of fibrinolysis. CONCLUSIONS: Beyond direct association of a PAI-2 polymorphism with recurrent risk in post-myocardial infarction patients, results suggest that PAI-2 likely plays a key role leading to risk through multiple pathophysiologic processes. Such knowledge could potentially be valuable with individualization of patient care.

Racial/Ethnic Difference in HIV-related Knowledge among Young Men who have Sex with Men and their Association with Condom Errors.

OBJECTIVE: HIV disproportionately affects young men who have sex with men, and knowledge about HIV transmission is one factor that may play a role in high rate of infections for this population. This study examined racial/ethnic differences in HIV knowledge among young men who have sex with men in the USA and its correlation to condom usage errors. DESIGN: Participants included an ethnically diverse sample of 344 young men who have sex with men screened from an ongoing longitudinal cohort study. Eligible participants were between the ages of 16 and 20 years, born male, and had previously had at least one sexual encounter with a man and/or identify as gay or bisexual. This analysis is based on cross-sectional data collected at the baseline interview using computer assisted self-interviewing (CASI) software. SETTING: Chicago, IL, USA. METHOD: We utilised descriptive and inferential statistics, including ANOVA and Tukey's Post hoc analysis to assess differences in HIV knowledge by level of education and race/ethnicity, and negative binomial regression to determine if HIV knowledge was associated with condom errors while controlling for age, education and race/ethnicity. RESULTS: The study found that Black men who have sex with men scored significantly lower (average score=67%; p<.05) than their White counterparts (average score=83%) on a measure of HIV knowledge (mean difference=16.1%, p<.001). Participants with less than a high school diploma and those with a high school diploma/GED only had lower knowledge scores, on average (66.4%, 69.9%, respectively) than participants who had obtained post-high school education (78.1%; mean difference=11.7%, 8.2% respectively, ps<.05). In addition, controlling for age, race and level of education, higher HIV knowledge scores were associated with fewer condom errors (Exp B =.995, CI 0.992-0.999, p<0.05). CONCLUSION: These findings stress the need to for increased attention to HIV transmission-related educational activities targeting the social realities and unique risk mechanisms of young men who have sex with men.

Insulin resistance predicts the risk for recurrent coronary events in post-infarction patients.

BACKGROUND: We investigated the risk for recurrent coronary events associated with insulin resistance in post-infarction patients from the Thrombogenic Factors and Recurrent Coronary Events (THROMBO) study. METHODS: The association between insulin resistance expressed by Homeostatic Model As-sessment 2 for Insulin Resistance (HOMA2-IR) and the risk for recurrent coronary events was investigated in a cohort of 1,032 patients evaluated 2 months after myocardial infarction (MI) with a follow-up of 26 months. The endpoint for the study was recurrent coronary event defined as cardiac death, nonfatal MI, or unstable angina, whichever occurred first. We used time dependent survival analysis and Cox proportional hazards regression method to determine the association between HOMA2 categorized as high > 75th percentile and endpoints after adjustment for relevant clinical covariates and series of thrombogenic and dyslipogenic factors. RESULTS: High HOMA2-IR defined as in fourth quartile (≥ 2.4) was associated with increased risk for recurrent coronary events (HR 1.44; CI 1.03-2.01; p = 0.03) after adjustment for the clinical covariates: age, gender, diabetes, prior MI, pulmonary congestion, coronary artery bypass grafting and percutaneous transluminal coronary angioplasty. The highest risk of cardiac events was observed in non-obese patients (body mass index [BMI] ≤ 30 kg/m2) with high HOMA2-IR (HR 1.5; CI 1.02-2.22; p = 0.038). The plasma level of plasminogen activa-tor inhibitor-1 was associated with higher risk for recurrent coronary events in patients with insulin resistance (HR 1.79; CI 1.05-3.03; p = 0.03, interaction p = 0.018). CONCLUSIONS: In conclusion, insulin resistance predicts recurrence of coronary events in post-infarction population. HOMA2-IR is better than BMI in stratifying risk of recurrent coronary events.

Plasminogen activator inhibitor-2 polymorphism associates with recurrent coronary event risk in patients with high HDL and C-reactive protein levels.

The objective of this work was to investigate whether fibrinolysis plays a role in establishing recurrent coronary event risk in a previously identified group of postinfarction patients. This group of patients was defined as having concurrently high levels of high-density lipoprotein cholesterol (HDL-C) and C-reactive protein (CRP) and was previously demonstrated to be at high-risk for recurrent coronary events. Potential risk associations of a genetic polymorphism of plasminogen activator inhibitor-2 (PAI-2) were probed as well as potential modulatory effects on such risk of a polymorphism of low-density lipoprotein receptor related protein (LRP-1), a scavenger receptor known to be involved in fibrinolysis in the context of cellular internalization of plasminogen activator/plansminogen activator inhibitor complexes. To this end, Cox multivariable modeling was performed as a function of genetic polymorphisms of PAI-2 (SERPINB, rs6095) and LRP-1 (LRP1, rs1800156) as well as a set of clinical parameters, blood biomarkers, and genetic polymorphisms previously demonstrated to be significantly and independently associated with risk in the study population including cholesteryl ester transfer protein (CETP, rs708272), p22phox (CYBA, rs4673), and thrombospondin-4 (THBS4, rs1866389). Risk association was demonstrated for the reference allele of the PAI-2 polymorphism (hazard ratio 0.41 per allele, 95% CI 0.20-0.84, p=0.014) along with continued significant risk associations for the p22phox and thrombospondin-4 polymorphisms. Additionally, further analysis revealed interaction of the LRP-1 and PAI-2 polymorphisms in generating differential risk that was illustrated using Kaplan-Meier survival analysis. We conclude from the study that fibrinolysis likely plays a role in establishing recurrent coronary risk in postinfarction patients with concurrently high levels of HDL-C and CRP as manifested by differential effects on risk by polymorphisms of several genes linked to key actions involved in the fibrinolytic process.

Optimal control in individual-based models: implications from aggregated methods.

Use of individual-based models (IBMs) has been expanding in both theoretical and applied ecology. IBMs include details at the level of individuals that may lead to different conclusions from aggregated modeling methods. There has been essentially no guidance available on how to most effectively manage populations when the underlying dynamics are best modeled through IBMs. Using a simple resource-consumer IBM, we investigate whether optimal control theory applied to an aggregated model (AM) can effectively control a harmful species modeled by an IBM or whether interactions between individuals, their spatial distribution, and/or landscape heterogeneities limit the effectiveness of a control derived for the AM. If optimal policies derived from an AM are determined to be generally effective in managing a population modeled with considerably greater complexity, this provides evidence that optimal management strategies may be relatively insensitive to the details of individual behavior and the associated effects on population response. We investigate these issues and find that if there is weak spatial heterogeneity in the resource, the optimal control derived from the AM can be used effectively to control the harmful species in the IBM. The approach is more limited in the case of very strong spatial heterogeneity in the resource. This suggests investigation of a mixture of simplified models in conjunction with detailed simulation models when individual differences affect population processes.

Thrombospondin-4 polymorphism (A387P) predicts cardiovascular risk in postinfarction patients with high HDL cholesterol and C-reactive protein levels.

Few studies are available in human populations investigating involvement of vascular inflammation and oxidative stress-related dysfunctional transformation of high-density lipoprotein (HDL) in establishing cardiovascular disease (CVD) risk. To this end, the current work investigated a subgroup of post-infarction patients at high-risk for recurrent events defined by high levels of HDL cholesterol (HDL-C) and concurrently high levels of C-reactive protein (CRP). Thrombospondin-4 (TSP-4), a matricellular protein of vessel walls associated with inflammation, was investigated in terms of CVD risk using multivariable modelling with a well-characterised functional genetic polymorphism of THBS4 (A387P, rs1866389) along with previously demonstrated risk-related functional genetic polymorphisms of CYBA (C242T, rs4673) and CETP (TaqIB, rs708272), and a set of blood markers. Results revealed risk-association for the gain-of-function P-allele of the THBS4 polymorphism (hazard ratio 2.00, 95% confidence interval 1.10-3.65, p=0.024). Additionally, von Willebrand factor was associated with D-dimer levels in the higher-risk P allele patients suggestive of a connection between endothelial dysfunction and thrombogenesis. In conclusion, TSP-4, a matricellular protein involved in regulating vascular inflammation, plays a role in establishing recurrent coronary risk in post-infarction patients with high levels of HDL-C and CRP. Further studies should focus on additional effects of vascular inflammatory processes on anti-atherogenic functionality of HDL particles.

Glycerol as a reference material for fecal fat quantitation using low-resolution time domain ¹H NMR spectroscopy.

OBJECTIVES: To assess glycerol as reference material for low-resolution time-domain (1)H NMR analysis of fecal fat. DESIGN AND METHODS: NMR analysis of fecal fat in stool samples with added glycerol was used to assess linearity, recovery, and relationship with NMR lipid signal. RESULTS: The study revealed for added glycerol excellent linearity (r=0.9998), recovery (101-104%), and linear relationship with simulated fecal fat content. CONCLUSIONS: Glycerol is an effective reference material for NMR fecal fat analysis.

Cholesteryl ester transfer protein polymorphism (TaqIB) associates with risk in postinfarction patients with high C-reactive protein and high-density lipoprotein cholesterol levels.

OBJECTIVE: To investigate the roles of inflammation and a cholesteryl ester transfer protein (CETP) polymorphism potentially related to recent findings demonstrating coronary risk with increasing high-density lipoprotein cholesterol (HDL-C) level. METHODS AND RESULTS: A novel graphical exploratory data analysis tool allowed the examination of coronary risk in postinfarction patients relating to HDL-C and C-reactive protein levels. Results demonstrated a high-risk subgroup, defined by high HDL-C and C-reactive protein levels, exhibiting larger HDL particles and lower lipoprotein-associated phospholipaseA(2) levels than lower-risk patients. Subgroup CETP-associated risk was probed using a functional CETP polymorphism (TaqIB, rs708272). In the high-risk subgroup, multivariable modeling revealed greater risk for B2 allele carriers (less CETP activity) versus B1 homozygotes (hazard ratio, 2.41; 95% CI, 1.04 to 5.60; P=0.04). Within the high-risk subgroup, B2 allele carriers had higher serum amyloid A levels than B1 homozygotes. Evidence also demonstrates that CETP genotypic differences in HDL subfraction distributions regarding non-HDL-C and lipoprotein-associated phospholipaseA(2) may potentially relate to impaired HDL remodeling. CONCLUSIONS: Postinfarction patients with high HDL-C and C-reactive protein levels demonstrate increased risk for recurrent events. Future studies should aim at characterizing altered HDL particles from such patients and at elucidating the mechanistic details related to inflammation and HDL particle remodeling. Such patients should be considered in drug trials involving an increase in HDL-C level.