Considerations for the Identification and Conveyance of Clinical Pathology Findings in Preclinical Toxicity Studies: Results From the 9th ESTP International Expert Workshop.

The European Society of Toxicologic Pathology (ESTP) organized a panel of 24 international experts from many fields of toxicologic clinical pathology (e.g., industry, academia, and regulatory) that came together in 2021 to align the use of terminology to convey the importance of clinical pathology findings in preclinical toxicity studies. An additional goal consisted of how to identify important findings in standard and nonstandard clinical pathology associated endpoints. This manuscript summarizes the information and opinions discussed and shared at the ninth ESTP International Expert Workshop, April 5 to 6, 2022. In addition to terminology usage, the workshop considered topics related to the identification and conveyance of the importance of test item-related findings. These topics included sources of variability, comparators, statistics, reporting, correlations to other study data, nonstandard biomarkers, indirect/secondary findings, and an overall weight-of-evidence approach.

Implementation of the Diabetes Prevention Program in Georgia Cooperative Extension According to RE-AIM and the Consolidated Framework for Implementation Research.

Increased dissemination of the CDC's Diabetes Prevention Program (DPP) is imperative to reduce type 2 diabetes. Due to its nationwide reach and mission to improve health, Cooperative Extension (Extension) is poised to be a sustainable DPP delivery system. However, research evaluating DPP implementation in Extension remains scant. Extension professionals delivered the DPP in a single-arm hybrid type II effectiveness-implementation study. Semi-structured interviews with Extension professionals were conducted at three time points. The Consolidated Framework for Implementation Research (CFIR) guided interview coding and analysis. Constructs were rated for magnitude and valence and evaluated as facilitators or barriers of RE-AIM (Reach, Effectiveness, Adoption, Implementation, Maintenance) outcomes. The program reached 119 participants, was adopted by 92% (n = 12/13) of trained Extension professionals and was implemented according to CDC standards: all programs exceeded the minimum 22-session requirement (26 ± 2 sessions). The program was effective in achieving weight loss (5.0 ± 5.2%) and physical activity (179 ± 122 min/week) goals. At post-intervention, eight professionals (67%) had begun or planned to maintain the intervention within the next 6 months. Several facilitators were identified, including Extension leadership structure, organizational compatibility, and technical assistance calls. Limited time to recruit participants was the primary barrier. Positive RE-AIM outcomes, facilitated by contextual factors, indicate Extension is an effective and sustainable DPP delivery system. Extension and other DPP implementers should plan strategies that promote communication, the program's evidence-base, recruitment time, and resource access. Researchers should explore DPP implementation in real-world settings to determine overall and setting-specific best practices, promote intervention uptake, and reduce diabetes.

Depressive symptoms, but not moderate-to-vigorous physical activity, impacts sexual well-being and menopause-specific quality of life in middle-aged women.

Inadequate physical activity (PA), unhealthy weight status, prevalence of chronic conditions, and psychosocial distress are common in middle-aged women and are linked to reductions in well-being and quality of life. However, their potential interactive effects, specifically on sexual well-being and menopause-specific quality of life (MENQOL), have not been well characterized in postmenopausal women. PURPOSE: To determine if moderate-to-vigorous physical activity (MVPA) and adiposity (%Fat) influence sexual well-being and MENQOL outcomes, controlling for health status (chronic conditions; medications) and psychosocial well-being (depressive symptoms; perceived stress), in postmenopausal women. Postmenopausal women (n = 68, 58.6 ± 3.4 yr, 80.9 percent married/partnered, 51.5 percent overweight/obese, nonsmoking) were recruited through e-mail advertisements and flyers placed throughout the community. Participants were scheduled for two laboratory visits 7-10 days apart where they were objectively assessed for MVPA with accelerometers (in the interim), adiposity via dual-energy X-ray absorptiometry (DXA), and self-report questionnaires to determine health status, depressive symptoms, perceived stress, sexual well-being, and MENQOL. Lower MVPA and higher %Fat were associated with lower physical domain MENQOL (both r = .27, p < .05); health status and psychosocial well-being were not associated (all p > .05). Hierarchical regression analyses revealed 1) greater number of chronic conditions and medications, and depressive symptoms scores predicted less favorable sexual well-being, independent of MVPA and %Fat (standardized ß range =.22-.56, all p < .05), 2) depression was most consistently associated with MENQOL (models p ≤ .001), and 3) greater adiposity augmented the negative influence of depression on the physical domain of MENQOL (ß = .40. p = .002). CONCLUSIONS: PA may influence sexual well-being and MENQOL indirectly through positive impacts on adiposity, chronic conditions, and depressive symptoms in middle-aged postmenopausal women, a sector of the population often afflicted with compromised sexual well-being.

Weight Loss and Exercise Effects on Rate of Torque Development and Physical Function in Overweight Older Women.

Exercise training (EX) and weight loss (WL) improve lower extremity physical function (LEPF) in older overweight women; however, effects on rate of torque development (RTD) are unknown. This study aimed to determine the effects of WL + EX or WL alone on RTD, and relatedly LEPF, in overweight older women. Leg strength was assessed using isokinetic dynamometry, and RTD was calculated (RTD200 = RTD at 200 ms, RTDPeak = peak RTD, T2P = time to 1st peak). LEPF was determined via clinical functional tasks. Women (n = 44, 69.1 ± 3.6 years, 30.6 ± 4.3 kg/m2) completed a 6-month trial in EX + WL or WL groups with similar weight loss (-9.8 ± 4.2%, p > .95). EX + WL had greater improvements in (a) most LEPF tasks (p < .001) and (b) RTD200, compared with WL (36% vs. -16%, p = .031); no other RTD parameters differed. Changes in RTD parameters and LEPF were not related (all p > .05). RTD is responsive to EX but is not associated with LEPF in older women.

Identifying support mechanisms to overcome barriers to food safety scheme certification in the food and drink manufacturing industry in Wales, UK.

Obtaining food safety certification is essential for food manufacturers. Potential barriers to obtaining certification are complex, interrelated and broadly relate to, 'knowledge and skills', 'time, cost and resources', and 'communication and access to information'. This study aimed to explore requirements for support to enable food manufacturers in Wales to overcome identified barriers. Food manufacturers (n = 37) participated in group discussions (n = 2) and completed online-questionnaires (n = 29). Support mechanisms, perceived necessary to obtain food safety certification included; funding for training and audit-fees, support for implementing food safety scheme documentation, on-site support through mentoring/coaching and pre-audits. Findings identify the need for a food safety scheme certification support package pathway incorporating online, off-site, on-site and financial support to assist food and drink manufacturers obtain third-party food safety certification. Such assistance would support three critical areas. Findings may inform development of support mechanisms to increase uptake of food safety certification and accelerate food-sector growth.

A case study of food handler hand hygiene compliance in high-care and high-risk food manufacturing environments using covert-observation.

Observation of behaviour is superior to cognitive data, which does not equate to behaviour. Covert-observation is seldom used in food manufacturing to assess behaviour. In this case study, closed-circuit-television footage (15 h) in a business were reviewed to assess hand hygiene compliance using an electronic-checklist. Hand hygiene attempts were observed prior to entering high-risk (cake/pie)(n= 47) and high-care (sandwich/salad)(n= 153) production areas. Business hand hygiene protocol required handwashing durations ≥ 20 s. Observed durations ranged 1-71 s, <96% of attempts were <20 s. Significantly longer durations were observed when food handlers were in the presence of others (12 s) than when alone (9 s). Although <99% utilised soap, only 56-69% wetted hands first. Failure to rub all parts of hands was commonplace (<87%) and 24-35% failed to apply sanitiser after drying. Consequently, >98% of observed attempts before entering production areas did not comply with the protocol. Observed non-compliant practices may have implications for food safety in manufacturing.

Understanding the barriers to food safety scheme certification in the food and drink manufacturing industry in Wales, UK.

It is increasingly essential for food manufacturers to have food safety certification. Little is known about certification in the Welsh food industry. This study aimed to explore perceptions of Welsh manufacturers regarding the drivers, benefits and barriers to obtaining certification. Focus groups with manufacturers and stakeholders (n = 68) were conducted. 'Customer requirement' and 'product safety' were drivers for obtaining certification. Benefits related to 'food safety culture', 'supply chain security', 'brand protection', 'due diligence', 'business growth' and 'job security'. Barriers were complex, often interrelated and were broadly defined as, 'knowledge and skills', 'time, cost and resources', and 'communication and access to information'. The research identifies the need to explore requirements for support to enable food manufacturers in Wales to overcome identified barriers. Such data may inform the design and development of support mechanisms to increase uptake of food safety certification and accelerate food sector growth in-line with Welsh Government aspirations.

Enzalutamide and Apalutamide: In Vitro Chemical Reactivity Studies and Activity in a Mouse Drug Allergy Model.

Enzalutamide and apalutamide are two androgen receptor inhibitors approved for the treatment of castration-resistant prostate cancer (CRPC) and nonmetastatic castration-resistant prostate cancer (nmCRPC), respectively. Apalutamide is associated with an increased incidence of skin rash above the placebo groups in the SPARTAN trial in nmCRPC and in the TITAN trial in metastatic castration-sensitive prostate cancer patients. On the contrary, the rate of skin rash across all clinical trials (including PROSPER [nmCRPC]) for enzalutamide is similar to the placebo. We hypothesized that the apalutamide-associated increased skin rash in patients could be linked to a structural difference. The 2-cyanophenyl and dimethyl moieties in enzalutamide are substituted in apalutamide with 2-cyanopyridine and cyclobutyl, respectively. In our evaluations, the 2-cyanopyridine moiety of apalutamide was chemically reactive with the thiol nucleophile glutathione, resulting in rearranged thiazoline products. Radiolabeled apalutamide, but not radiolabeled enzalutamide, was shown to react with mouse and human plasma proteins. Thiol nucleophiles decreased the extent of covalent binding to the model protein bovine serum albumin, whereas amine and alcohol nucleophiles had no effect, suggesting reactivity with cysteine of proteins. Subcutaneous administration of apalutamide dose dependently increased lymphocyte cellularity in draining lymph nodes in a mouse drug allergy model (MDAM). Enzalutamide, and its known analogue RD162 in which the cyanophenyl was retained but the dimethyl was replaced by cyclobutyl, demonstrated substantially less covalent binding activity and negative results in the MDAM assay. Collectively, these data support the hypothesis that the 2-cyanopyridine moiety in apalutamide may react with cysteine in proteins forming haptens, which may trigger an immune response, as indicated by the activity of apalutamide in the MDAM assay, which in turn may be leading to increased potential for skin rash versus placebo in patients in the SPARTAN and TITAN clinical trials.

Video observation of hand-hygiene compliance in a manufacturer of ready-to-eat pie and pastry products.

Food-handler hand-hygiene can be a contributory factor for food-borne illness. Cognitive data (knowledge/attitudes/self-reported practices), while informative, are not indicative of behaviour, and are subject to biases. Consequently, observation of behaviour is superior to survey data. However, researcher presence in direct observation increases reactivity, whereas video observation gives comprehensive analysis over a longer period, furthermore, familiarity reduces reactivity. Although video observation has been used to assess food safety at retail/foodservice, this valuable method is under-utilized in food-manufacturing environments. For the study, footage (24 h) was reviewed to assess compliance in a food-manufacturing site with company protocol. Video observation of food-handlers entering production (n = 674) was assessed; upon 70 occasions no attempt to implement hand-hygiene was observed. Of attempted hand-hygiene practices (n = 604), only 2% implemented compliant practices. Although 78% of attempts utilized soap, only 42% included sanitizer. Duration ranged from 1 to 69 s (Median 17 s). The study provides hand-hygiene data in an area that observational data is seldom captured.

Moderate, but not vigorous, intensity exercise training reduces C-reactive protein.

BACKGROUND: Sprint interval cycle training is a contemporary popular mode of training but its relative efficacy, under conditions of matched energy expenditure, to reduce risk factors for cardiometabolic disease is incompletely characterised, especially in young women. The purpose of this investigation was to determine the relative efficacy of six weeks of moderate-intensity cycling (MOD-C) and vigorous sprint-interval cycling (VIG-SIC) on lipid profile, insulin (INS) and insulin resistance using the homeostatic model assessment (HOMA-IR) and C-reactive protein (CRP) in inactive, overweight/obese (OW/OB) young women. METHODS: Participants (BMI ≥25 kg/m2, waist circumference ≥88 cm) were randomly assigned to MOD-C (20-30 min at 60-70% of heart rate reserve(HRR)) or VIG-SIC (5-7 repeated bouts 30-second maximal effort sprints, followed by four minutes of active recovery) supervised training three days/week for six weeks, with each group matched on energy expenditure. Adiposity (%Fat) was measured using dual x-ray absorptiometry. RESULTS: Forty-four participants (20.4 ± 1.6 years, 65.9% Caucasian, 29.8 ± 4.1 kg/m2) were included in the analysis. The improvement in CRP observed in the MOD-C group was larger than the VIG-C group (p = .034). Overall, high-density lipoprotein (HDL-C) and low-density lipoprotein (LDL-C) levels improved following training (p < .05); however, total cholesterol, triglyceride, INS and HOMA-IR did not improve (p > .05). CONCLUSION: These results indicate MOD-C training may be more effective in reducing CRP than VIG-SIC.

Interactive associations of physical activity, adiposity, and oral contraceptive use on C-reactive protein levels in young women.

Oral contraceptives (OCs) are the most frequently used type of birth control among young women. OC-users have higher C-reactive protein (CRP) values, an indicator of systemic inflammation, than do non-OC-users. In addition, adiposity (percent fat) is positively associated with CRP, and physical activity (PA) is inversely associated with CRP. The present study determined the interactive associations of PA, percent fat, and OC-use with CRP. Data were collected during 2012-2015 at the University of Georgia. Objective PA was measured via pedometers. Percent fat was measured via dual X-ray absorptiometry. The current OC-use was self-reported. High-sensitivity (hs) CRP was determined using venipuncture. Multivariate linear regression determined the interactive associations of percent fat, OC-use, and PA with hs-CRP. Participants (n = 247; mean age 18.9 ± 1.4 years, 60.7 percent white) accumulated a mean of 10,075.7 ± 3,593.4 steps/day. One-third of participants were categorized as overweight/obese by BMI (mean = 24.5 ± 4.8 kg/m2, mean percent fat = 35.2 ± 6.8). The current OC-use was reported by 26.2 percent of the sample (n = 61). A significant three-way interaction (ß = 0.01, p = .03) indicated that higher PA was associated with lower hs-CRP in non-OC-users with higher percent fat, but not among OC-users with higher percent fat. These results highlight the need to measure and account for the current OC-use in studies examining the relationship between PA and CRP.

Moderate-intensity physical activity is independently associated with lower-extremity muscle power in older women.

Skeletal muscle power is a salient determinant of physical function in older adults, but its relationship with habitual physical activity has not been well-characterized. The aim of this study was to examine the association between moderate-intensity physical activity and lower-extremity muscle power in community-dwelling older women. Older women (n = 96, mean age = 73.9 ± 5.6 years, mean body mass index = 26.5 ± 4.7 kg/m(2)) underwent assessments for body composition via dual-energy X-ray absorptiometry and lower-extremity muscle power (watts) using the Nottingham power rig. The Community Health Activities Model Program for Seniors questionnaire was used to estimate weekly caloric expenditure in moderate-intensity physical activity (kcals/wk). Linear regression indicated that moderate-intensity physical activity was independently related to muscle power (standardized ß = 0.20, p = .03), and this relationship remained following adjustment for covariates. Analysis of covariance revealed that women in the highest tertile of volume of physical activity had significantly greater muscle power than those with the lowest volume (199.0 vs. 170.7 watts, p < .05). Moderate-intensity physical activity was independently associated with lower-extremity muscle power in older women. Future intervention trials should determine if increasing habitual physical activity is associated with improvements in lower-extremity muscle power in older women.

The negative association of childhood obesity to cognitive control of action monitoring.

The global epidemic of childhood obesity has become a major public health concern. Yet, evidence regarding the association between childhood obesity and cognitive health has remained scarce. This study examined the relationship between obesity and cognitive control using neuroelectric and behavioral measures of action monitoring in preadolescent children. Healthy weight and obese children performed compatible and incompatible stimulus-response conditions of a modified flanker task, while task performance and the error-related negativity (ERN) were assessed. Analyses revealed that obese children exhibited a longer reaction time (RT) relative to healthy weight children for the incompatible condition, whereas no such difference was observed for the compatible condition. Further, obese children had smaller ERN amplitude relative to healthy weight children with lower post-error response accuracy. In addition, healthy weight children maintained post-error response accuracy between the compatible and incompatible conditions with decreased ERN amplitude in the incompatible condition, whereas obese children exhibited lower post-error response accuracy for the incompatible relative to the compatible condition with no change in ERN amplitude between the compatibility conditions. These results suggest that childhood obesity is associated with a decreased ability to modulate the cognitive control network, involving the prefrontal cortex and anterior cingulate cortex, which supports action monitoring.

Regulatory forum commentary: is unexpected immunostimulation manageable in pharmaceutical development?

One of the biggest challenges in drug development is interpretation of findings suggestive of immunostimulation, particularly when the findings are unexpected based on the known mechanism of action of the drug. Findings suggestive of immunostimulation do not necessarily preclude development of the drug. By understanding mechanism, the patient population, and species differences and by careful clinical monitoring, many of the drugs that produce immunostimulation can be successfully developed. This opinion piece describes various types of immunostimulation and a risk assessment strategy for management of unexpected immunostimulation in toxicity studies.

Approaches and considerations for the assessment of immunotoxicity for environmental chemicals: a workshop summary.

As experience is gained with toxicology testing and as new assays and technologies are developed, it is critical for stakeholders to discuss opportunities to advance our overall testing strategies. To facilitate these discussions, a workshop on practices for assessing immunotoxicity for environmental chemicals was held with the goal of sharing perspectives on immunotoxicity testing strategies and experiences, developmental immunotoxicity (DIT), and integrated and alternative approaches to immunotoxicity testing. Experiences across the chemical and pharmaceutical industries suggested that standard toxicity studies, combined with triggered-based testing approaches, represent an effective and efficient approach to evaluate immunotoxic potential. Additionally, discussions on study design, critical windows, and new guideline approaches and experiences identified important factors to consider before initiating DIT evaluations including assay choice and timing and the impact of existing adult data. Participants agreed that integrating endpoints into standard repeat-dose studies should be considered for fulfilling any immunotoxicity testing requirements, while also maximizing information and reducing animal use. Participants also acknowledged that in vitro evaluation of immunosuppression is complex and may require the use of multiple assays that are still being developed. These workshop discussions should contribute to developing an effective but more resource and animal efficient approach for evaluating chemical immunotoxicity.

V. The differential association of adiposity and fitness with cognitive control in preadolescent children.

With the increasing prevalence of sedentary behaviors during childhood, a greater understanding of the extent to which excess adiposity and aerobic fitness relate to cognitive health is of increasing importance. To date, however, the vast majority of research in this area has focused on adiposity or fitness, rather than the possible inter-relationship, as it relates to cognition. Accordingly, this study examined the differential associations between body composition, aerobic fitness, and cognitive control in a sample of 204 (96 female) preadolescent children. Participants completed a modified flanker task (i.e., inhibition) and a switch task (i.e., cognitive flexibility) to assess two aspects of cognitive control. Findings from this study indicate that fitness and adiposity appear to be separable factors as they relate to cognitive control, given that the interaction of fitness and adiposity was observed to be nonsignificant for both the flanker and switch tasks. Fitness exhibited an independent association with both inhibition and cognitive flexibility whereas adiposity exhibited an independent association only with cognitive flexibility. These results suggest that while childhood obesity and fitness appear to both be related to cognitive control, they may be differentially associated with its component processes.

Area-level socioeconomic status, adiposity, physical activity, and inflammation in young adults, 2013.

INTRODUCTION: We assessed the independent effects of socioeconomic status, sex, adiposity, and physical activity on C-reactive protein in young adults. METHODS: During the fall semester of their first year, college students (n = 177; mean age, 18.1 y; 66.7% female; 65.5% white) were assessed for adiposity via dual x-ray absorptiometry, physical activity via accelerometer, and serum C-reactive protein. Area-level socioeconomic status was based on self-reported home zip code. Hierarchical linear modeling was used to assess the relationship of sex, adiposity, and physical activity on the dependent variable of C-reactive protein, with participants nested within geographic regions of similar socioeconomic characteristics. RESULTS: C-reactive protein was positively associated with adiposity and inversely associated with socioeconomic status (both P < .05). Area-level socioeconomic status explained 28.2% of the variance in C-reactive protein. Adiposity was significantly associated with C-reactive protein in the full model (P = .006); physical activity was not associated with C-reactive protein (P = .48), and area-level socioeconomic status approached significance (P = .05) within the age range of our analysis after accounting for the variance explained by adiposity. CONCLUSION: The significant positive association between adiposity and C-reactive protein suggests that young adults with higher adiposity have higher C-reactive protein levels after accounting for area-level socioeconomic status, sex, and physical activity.

Greater effect of adiposity than physical activity or lean mass on physical function in community-dwelling older adults.

Adiposity, lean mass, and physical activity (PA) are known to influence physical function in older adults, although the independent influences are not completely characterized. Older adults (N = 156, M age = 68.9 ± 6.7 yr, 85 men) were assessed for body composition via dual-energy X-ray absorptiometry, PA by accelerometer, and physical function via timed up-and-go (UP&GO), 30-s chair stand, 6-min walk (6-min WALK), and Star-Excursion Balance Test. In the absence of percentage-body-fat by PA interactions (p > .05), main effects existed such that a higher percentage body fat was associated with poorer performance in UP&GO, 30-s chair stand, and 6-min WALK (p < .05). No significant main effects were found for PA and functional performance. Adiposity explains 4.6-11.4% in physical functional variance (p < .05). Preventing increases in adiposity with age may help older adults maintain functional independence.