Fifteen day repeat air: liquid Interface air-only exposures can cause respiratory epithelium injury in MucilAir™ nasal respiratory epithelial cells that parallels chemically induced cytotoxicity.

New Approach Methodologies (NAMs) are being widely used to reduce, refine, and replace, animal use in studying toxicology. For respiratory toxicology, this includes in silico and in vitro alternatives using air:liquid interface (ALI) exposures to replace traditional in vivo inhalation studies. In previous studies using 1,3-dichloropropene (1,3-DCP), a 5-day 4 h repeat exposures of MucilAir™ nasal cell culture models caused, dose-dependent cytotoxicity, depletion of GSH, changes in differential gene expression and histopathological transitions in cellular morphology from pseudostratified columnar epithelium to squamous epithelium. In this report we attempted to extend these studies using 15-day 1,3-DCP 4 h exposures to using MucilAir™ nasal cultures as outlined by an US EPA recent task order (US EPA 2023). For the 15-day repeat exposure, there were severe histopathologic changes in the MucilAir™ nasal mock-treatment (air-only) VITROCELL® chamber controls compared to incubator controls preventing any further analysis. The histopathological transitions in cellular morphology from pseudostratified columnar epithelium to squamous epithelium observed in the air only control in this study and previously with 1,3-DCP in MucilAir™ nasal cultures is also a hallmark of chemically induced cytotoxic responses in vivo in the respiratory tract. Histopathology assessments of 3D respiratory tract models used in ALI exposures can provide the linkage between in vitro to in vivo outcomes as part of the validation efforts of ALI use in regulatory toxicology. This report indicates that importance of histopathological assessments of incubator and mock-treatment (air-only) controls from each ALI exposure experiment along with exposed cell based model.

From first encounters to longitudinal exposure: a repeated exposure-test paradigm for monitoring speech adaptation.

Perceptual difficulty with an unfamiliar accent can dissipate within short time scales (e.g., within minutes), reflecting rapid adaptation effects. At the same time, long-term familiarity with an accent is also known to yield stable perceptual benefits. However, whether the long-term effects reflect sustained, cumulative progression from shorter-term adaptation remains unknown. To fill this gap, we developed a web-based, repeated exposure-test paradigm. In this paradigm, short test blocks alternate with exposure blocks, and this exposure-test sequence is repeated multiple times. This design allows for the testing of adaptive speech perception both (a) within the first moments of encountering an unfamiliar accent and (b) over longer time scales such as days and weeks. In addition, we used a Bayesian ideal observer approach to select natural speech stimuli that increase the statistical power to detect adaptation. The current report presents results from a first application of this paradigm, investigating changes in the recognition accuracy of Mandarin-accented speech by native English listeners over five sessions spanning 3 weeks. We found that the recognition of an accent feature (a syllable-final /d/, as in feed, sounding/t/-like) improved steadily over the three-week period. Unexpectedly, however, the improvement was seen with or without exposure to the accent. We discuss possible reasons for this result and implications for conducting future longitudinal studies with repeated exposure and testing.

Efficacy of melatonin in alleviating disorders arising from repeated exposure to sevoflurane in males and females of the Wistar rats during preadolescence.

Pediatricians use sevoflurane due to its fast action and short recovery time. However, studies have shown that repeated exposure to anesthesia can affect learning and memory. Melatonin, an indole-type neuroendocrine hormone, has significant anti-inflammatory, and neuroprotective properties. Melatonin's impact on cognitive behavior in sevoflurane-anesthetized males and females of the Wistar rats during preadolescence was examined in this research. The cognitive function was evaluated by shuttle box and morris water maze tests, while interleukin-10, Catalase (CAT), Malondialdehyde (MDA), and Tumor Necrosis Factor-α (TNF-α) were evaluated using ELISA kits. The expression levels of the apoptosis-linked proteins, Bax, Bcl-2, and caspase-3, were determined using the western blotting technique. The learning and memory latencies of the rats were more significant in the sevoflurane groups than in the control group; however, the latencies were significantly shorter in the sevoflurane and melatonin groups than in the control group. The levels of MDA, TNF-α, Bax, and caspase-3 were significantly higher in the sevoflurane groups than in the control group. We also found that the levels of CAT and Bcl-2 were significantly reduced in the sevoflurane groups compared to the control group. Increasing levels of CAT, Bcl-2, and decreasing levels of MDA, TNF-α, Bax, and caspase-3 in response to melatonin indicate a possible contribution to the recovery from the sevoflurane impairment. Melatonin shows neuroprotective effects in male and female rats with sevoflurane-induced cognitive impairment. This suggests melatonin could be a valuable treatment for learning and memory deficits resulting from repeated exposure to sevoflurane, possibly by controlling apoptosis, oxidative stress, and inflammation.

Haptoglobin Attenuates Cerebrospinal Fluid Hemoglobin-Induced Neurological Deterioration in Sheep.

Secondary brain injury (SBI) occurs with a lag of several days post-bleeding in patients with aneurysmal subarachnoid hemorrhage (aSAH) and is a strong contributor to mortality and long-term morbidity. aSAH-SBI coincides with cell-free hemoglobin (Hb) release into the cerebrospinal fluid. This temporal association and convincing pathophysiological concepts suggest that CSF-Hb could be a targetable trigger of SBI. However, sparse experimental evidence for Hb's neurotoxicity in vivo defines a significant research gap for clinical translation. We modeled the CSF-Hb exposure observed in aSAH patients in conscious sheep, which allowed us to assess neurological functions in a gyrencephalic species. Twelve animals were randomly assigned for 3-day bi-daily intracerebroventricular (ICV) injections of either Hb or Hb combined with the high-affinity Hb scavenger protein haptoglobin (Hb-Hp, CSL888). Repeated CSF sampling confirmed clinically relevant CSF-Hb concentrations. This prolonged CSF-Hb exposure over 3 days resulted in disturbed movement activity, reduced food intake, and impaired observational neuroscores. The Hb-induced neurotoxic effects were significantly attenuated when Hb was administered with equimolar haptoglobin. Preterminal magnetic resonance imaging (MRI) showed no CSF-Hb-specific structural brain alterations. In both groups, histology demonstrated an inflammatory response and revealed enhanced perivascular histiocytic infiltrates in the Hb-Hp group, indicative of adaptive mechanisms. Heme exposure in CSF and iron deposition in the brain were comparable, suggesting comparable clearance efficiency of Hb and Hb-haptoglobin complexes from the intracranial compartment. We identified a neurological phenotype of CSF-Hb toxicity in conscious sheep, which is rather due to neurovascular dysfunction than structural brain injury. Haptoglobin was effective at attenuating CSF-Hb-induced neurological deterioration, supporting its therapeutic potential.

Low-concentration repeated exposure and high-concentration single exposure of cyanamide suppressed the growth of redroot pigweed in the alfalfa field.

The inclination toward natural products has led to the onset of the discovery of new bioactive metabolites that could be targeted for specific therapeutic or agronomic applications. Despite increasing knowledge coming to light of plant-derived materials as leads for new herbicides, relatively little is known about the mode of action on herbicide-resistant weeds. Cyanamide (CA) is a naturally occurring herbicide synthesized by hairy vetch (Vicia villosa Roth.). However, it has not been experimentally verified whether CA suppresses target plants via sustained discharge at low concentrations, as is often the case with most plant-derived materials. This study aimed to detect the toxicity and the mode of action of CA to alfalfa (Medicago sativa L.) and redroot pigweed (Amaranthus retroflexus L.). The toxicity of CA toward the alfalfa and redroot pigweed by three different exposure patterns was compared: low-concentration repeated exposure with 0.3 g/L CA (LRE), high-concentration single exposure with 1.2 g/L CA (HSE), and distilled water spray as control. The results showed that CA had a stronger inhibitory effect on redroot pigweed growth compared to alfalfa under both LRE and HSE exposure modes, with leaves gradually turning yellow and finally wilting. Beyond that, field trials were conducted to corroborate the toxicity of CA to alfalfa and redroot pigweed. The results have also shown that CA could inhibit the growth of redroot pigweed without significant adverse effects on alfalfa. The outcomes concerning electrolyte permeability, root activity, and malondialdehyde (MDA) content indicated that CA suppressed the growth of redroot pigweed by interfering with the structure of the cell membrane and impacting cellular osmotic potential. CA could destroy the cell membrane structure to inhibit the growth of the redroot pigweed by both LRE and HSE exposure modes, which provides a theoretical basis for preventing and controlling redroot pigweed in alfalfa fields.

The health risks of marine biotoxins associated with high seafood consumption: Looking beyond the single dose, single outcome paradigm with a view towards addressing the needs of coastal Indigenous populations in British Columbia.

People who consume high quantities of seafood are at a heightened risk for marine biotoxin exposure. Coastal Indigenous peoples may experience higher levels of risk than the general population due to their reliance on traditional marine foods. Most evidence on the health risks associated with biotoxins focus on a single exposure at one point in time. There is limited research on other types of exposures that may occur among those who regularly consume large quantities of seafood. The objective of this review is to assess what is known about the unique biotoxin exposure risks associated with the consumption patterns of many coastal Indigenous populations. These risks include [1]: repeated exposure to low doses of a single or multiple biotoxins [2]; repeated exposures to high doses of a single or multiple biotoxins; and [3] exposure to multiple biotoxins at a single point in time. We performed a literature search and collected 23 recent review articles on the human health effects of different biotoxins. Using a narrative framework synthesis approach, we collated what is known about the health effects of the exposure risks associated with the putative consumption patterns of coastal Indigenous populations. We found that the health effects of repeated low- or high-dose exposures and the chronic health effects of marine biotoxins are rarely studied or documented. There are gaps in our understanding of how risks differ by seafood species and preparation, cooking, and consumption practices. Together, these gaps contribute to a relatively poor understanding of how biotoxins impact the health of those who regularly consume large quantities of seafood. In the context of this uncertainty, we explore how known and potential risks associated with biotoxins can be mitigated, with special attention to coastal Indigenous populations routinely consuming seafood. Overall, we conclude that there is a need to move beyond the single-dose single-outcome model of exposure to better serve Indigenous communities and others who consume high quantities of seafood.

Effect of cell treatment procedures on in vitro genotoxicity assessment.

So far, the majority of in vitro toxicological experiments are conducted after an acute 24 h treatment that does not represent a realistic human chemical exposure. Recently, new in vitro approaches have been proposed to study the chemical toxicological effect over several days in order to be more predictive of a representative exposure scenario. In this study, we investigated the genotoxic potential of chemicals (direct or bioactived clastogen, aneugen and apoptotic inducer) with the γH2AX and pH3 biomarkers, in the human liver-derived HepaRP cell line. We used different treatment durations, with or without a three-day recovery stage (release period), before genotoxicity measurement. Data were analysed with the Benchmark Dose approach. We observed that the detection of clastogenic compounds (notably for DNA damaging agents) was more sensitive after three days of repeated treatment compared to one or three treatments over 24 h. In contrast, aneugenic chemicals were detected as genotoxic in a similar manner whether after a 24 h exposure or a three-day repeated treatment. Globally, the release period decreases the genotoxicity measurement substantially. For DNA damaging agents, after high concentration treatments, γH2AX induction was always observed after a three-day release period. In contrast, for DNA topoisomerase inhibitors, no effect could be observed after the release period. In conclusion, in the HepaRP cell line, there are some important differences between a one-day acute and a three-day repeated treatment protocol, indicating that different cell treatment procedures may differentiate chemical genotoxic mechanisms of action more efficiently.

I'll Never Give Up: A Qualitative Study of Caregivers' Perceptions and Decisional Processes When Feeding Infants and Toddlers Novel and Disliked Foods.

OBJECTIVE: To better understand caregivers' decisional processes related to offering novel and disliked foods to their infants and toddlers. DESIGN: As part of a parent study on young children's food acceptance that took place in Denver, CO, this secondary analysis used a basic qualitative approach to explore caregivers' decisional processes related to repeated exposure and children's food rejection. PARTICIPANTS: English-speaking caregivers of infants and toddlers (aged 6-24 months; n = 106) were recruited via flyers and social media and interviewed (from July, 2017 to January, 2018) during a laboratory visit focused on introducing a novel food. PHENOMENON OF INTEREST: Factors influencing caregiver decisions to (dis)continue offering novel or disliked foods. ANALYSIS: Using a combined deductive and inductive coding approach, trained researchers coded transcripts and codes, which were reviewed and discussed by all investigators to identify themes. RESULTS: Three major themes (and 2 subthemes) were generated regarding caregivers' decisions about re-offering rejected foods: 1) Caregivers understand that multiple experiences with new foods are needed because children's reactions can be unpredictable and depend upon time, developmental stage, and child traits; 2) Caregivers vary in their persistence and decisions to keep offering foods depending on responsiveness to child cues (sub-theme) and adult-centered beliefs, needs, and decisions (sub-theme); 3) Child food acceptance will change with time, circumstances, and development if you keep trying. CONCLUSIONS AND IMPLICATIONS: Although caregivers are aware of repeated exposure, additional implementation research focused on translating theory into effective home practices could assist caregivers to persist in offering novel or disliked foods.

Repeated exposure to body-related memories in women with high body-related self-disgust: Impact on disgust, avoidance, and acceptance.

Disgust-based body image concerns can bias autobiographical memory towards the recall and avoidant processing of disgust-related memories of the own body. Repeated exposure to such memories may help breaking avoidance and promote the habituation of disgust, thereby lowering body concerns. Using a pre-post within-participant experimental design, we tested if repeatedly exposing women with high self-disgust (N = 61) to disgust-focused body memories vs. neutral memories led to changes in disgust, body acceptance, and reactive avoidance. Contrary to expectations, state disgust towards the body itself only decreased following exposure to neutral memories. Yet, disgust elicited by body-related memories decreased following both repeated exposure to neutral and body memories. Although acceptance was not found to significantly change in either exposure session, pre-post decreases in state disgust were associated with increases in acceptance following the disgust-focused exposure. In contrast to expectations, reactive avoidance increased from pre to post in the disgust-focused exposure. Overall, the results indicate that repeated exposure to disgust-focused body memories may help reduce disgust elicited by these memories and promote body acceptance. Yet, the effect of this repeated exposure to body memories did not extend to changing state disgust towards the body, possibly due to reactive avoidance.

Assessment of brain response in operators subject to recoil force from firing long-range rifles.

Mild traumatic brain injury (mTBI) may be caused by occupational hazards military personnel encounter, such as falls, shocks, exposure to blast overpressure events, and recoil from weapon firing. While it is important to protect against injurious head impacts, the repeated exposure of Canadian Armed Forces (CAF) service members to sub-concussive events during the course of their service may lead to a significant reduction in quality of life. Symptoms may include headaches, difficulty concentrating, and noise sensitivity, impacting how personnel complete their duties and causing chronic health issues. This study investigates how the exposure to the recoil force of long-range rifles results in head motion and brain deformation. Direct measurements of head kinematics of a controlled population of military personnel during firing events were obtained using instrumented mouthguards. The experimentally measured head kinematics were then used as inputs to a finite element (FE) head model to quantify the brain strains observed during each firing event. The efficacy of a concept recoil mitigation system (RMS), designed to mitigate loads applied to the operators was quantified, and the RMS resulted in lower loading to the operators. The outcomes of this study provide valuable insights into the magnitudes of head kinematics observed when firing long-range rifles, and a methodology to quantify effects, which in turn will help craft exposure guidelines, guide training to mitigate the risk of injury, and improve the quality of lives of current and future CAF service members and veterans.

Repeated Exposure Enhanced Toxicity of Clarithromycin on Microcystis aeruginosa Versus Single Exposure through Photosynthesis, Oxidative Stress, and Energy Metabolism Shift.

Antibiotics are being increasingly detected in aquatic environments, and their potential ecological risk is of great concern. However, most antibiotic toxicity studies involve single-exposure experiments. Herein, we studied the effects and mechanisms of repeated versus single clarithromycin (CLA) exposure on Microcystis aeruginosa. The 96 h effective concentration of CLA was 13.37 µg/L upon single exposure but it reduced to 6.90 µg/L upon repeated exposure. Single-exposure CLA inhibited algal photosynthesis by disrupting energy absorption, dissipation and trapping, reaction center activation, and electron transport, thereby inducing oxidative stress and ultrastructural damage. In addition, CLA upregulated glycolysis, pyruvate metabolism, and the tricarboxylic acid cycle. Repeated exposure caused stronger inhibition of algal growth via altering photosynthetic pigments, reaction center subunits biosynthesis, and electron transport, thereby inducing more substantial oxidative damage. Furthermore, repeated exposure reduced carbohydrate utilization by blocking the pentose phosphate pathway, consequently altering the characteristics of extracellular polymeric substances and eventually impairing the defense mechanisms of M. aeruginosa. Risk quotients calculated from repeated exposure were higher than 1, indicating significant ecological risks. This study elucidated the strong influence of repeated antibiotic exposure on algae, providing new insight into antibiotic risk assessment.

What's cooking, if not meat? Effects of repeated home-use, recipe inspiration and meal context on perception of plant-based meat analogues.

Plant-based meat analogues (PBMA) may help consumers in shifting towards more plant-based diets, but PBMA are not widely used yet, and little is known about their longer-term acceptance. This study investigated whether consumer acceptance of PBMA changed with repeated home-use, and whether providing recipe suggestions in the form of meal boxes could influence PBMA acceptance. To this end, Dutch regular meat eaters (n = 61) prepared, consumed and evaluated two meals (one from a meal box, one self-created) with PBMA (PB mince and PB chicken, counterbalanced across meal types) per week at home for four weeks. As a secondary objective, potential longer-term effects of repeated home-use of PBMA on meat (analogue) consumption habits and attitudes (e.g. motives for choosing PBMA, attitudes toward eating less meat) were assessed in a pre-vs post-intervention survey. Responses were compared with a control group of consumers not participating in the home-use study (n = 179). Results provided no evidence that PBMA liking changed with repeated home-use, nor that the provision of meal boxes increased liking of PBMA. Instead, PBMA liking was strongly influenced by the meal context, which may have overruled potential effects of repeated exposure. Findings from the pre- vs. post-intervention survey suggest that repeated exposure may stimulate longer-term consumption of PBMA, although more seems needed to bring about a structural shift toward a less animal-based consumption pattern. Future research should investigate whether more sophisticated recipes that provide a suitable meal context for PBMA and elevate consumers' meal experiences may improve PBMA acceptance and facilitate the transition toward more sustainable diets.

Mutagenic effects of a commercial glyphosate-based herbicide formulation on the soil filamentous fungus Aspergillus nidulans depending on the mode of exposure.

Glyphosate-based herbicides (GBH) are the most used pesticides worldwide. This widespread dissemination raises the question of non-target effects on a wide range of organisms, including soil micro-organisms. Despite a large body of scientific studies reporting the harmful effects of GBHs, the health and environmental safety of glyphosate and its commercial formulations remains controversial. In particular, contradictory results have been obtained on the possible genotoxicity of these herbicides depending on the organisms or biological systems tested, the modes and durations of exposure and the sensitivity of the detection technique used. We previously showed that the well-characterized soil filamentous fungus Aspergillus nidulans was highly affected by a commercial GBH formulation containing 450 g/L of glyphosate (R450), even when used at doses far below the agricultural application rate. In the present study, we analysed the possible mutagenicity of R450 in A. nidulans by screening for specific mutants after different modes of exposure to the herbicide. R450 was found to exert a mutagenic effect only after repeated exposure during growth on agar-medium, and depending on the metabolic status of the tested strain. The nature of some mutants and their ability to tolerate the herbicide better than did the wild-type strain suggested that their emergence may reflect an adaptive response of the fungus to offset the herbicide effects. The use of a non-selective molecular approach, the quantitative random amplified polymorphic DNA (RAPD-qPCR), showed that R450 could also exert a mutagenic effect after a one-shot overnight exposure during growth in liquid culture. However, this effect was subtle and no longer detectable when the fungus had previously been repeatedly exposed to the herbicide on a solid medium. This indicated an elevation of the sensitivity threshold of A. nidulans to the R450 mutagenicity, and thus confirmed the adaptive capacity of the fungus to the herbicide.

Increasing confidence in new approach methodologies for inhalation risk assessment with multiple end point assays using 5-day repeated exposure to 1,3-dichloropropene.

New Approach Methodologies (NAMs) are being widely used to reduce, refine, and replace, animal use in studying toxicology. For respiratory toxicology, this includes both in silico and in vitro alternatives to replace traditional in vivo inhalation studies. 1,3-Dichloropropene (1,3-DCP) is a volatile organic compound that is widely used in agriculture as a pre-planting fumigant. Short-term exposure of humans to 1,3-DCP can result in mucous membrane irritation, chest pain, headache, and dizziness. In our previous work, we exposed differentiated cells representing different parts of the respiratory epithelium to 1,3-DCP vapor, measured cytotoxicity, and did In Vitro to In Vivo Extrapolation (IVIVE). We have extended our previous study with 1,3-DCP vapors by conducting transcriptomics on acutely exposed nasal cultures and have implemented a separate 5-day repeated exposure with multiple endpoints to gain further molecular insight into our model. MucilAir™ Nasal cell culture models, representing the nasal epithelium, were exposed to six sub-cytotoxic concentrations of 1,3-DCP vapor at the air-liquid interface, and the nasal cultures were analyzed by different methodologies, including histology, transcriptomics, and glutathione (GSH) -depletion assays. We observed the dose-dependent effect of 1,3-DCP in terms of differential gene expression, change in cellular morphology from pseudostratified columnar epithelium to squamous epithelium, and depletion of GSH in MucilAir™ nasal cultures. The MucilAir™ nasal cultures were also exposed to 3 concentrations of 1,3-DCP using repeated exposure 4 h per day for 5 days and the histological analyses indicated changes in cellular morphology and a decrease in ciliated bodies and an increase in apoptotic bodies, with increasing concentrations of 1,3-DCP. Altogether, our results suggest that sub-cytotoxic exposures to 1,3-DCP lead to several molecular and cellular perturbations, providing significant insight into the mode-of-action (MoA) of 1,3-DCP using an innovative NAM model.

Novel nasal formulation of xylometazoline with hyaluronic acid: In vitro ciliary beat frequency study.

Acute viral rhinosinusitis (viral ARS), or commonly referred to as the "common cold", is caused by respiratory viruses that cause disruption of the airway epithelial barrier and mucociliary dysfunction. Treatment of ARS is mainly symptomatic, with xylometazoline, a direct-acting α-adrenoceptor agonist, commonly used as a nasal decongestant. Unfortunately, this treatment does not resolve the epithelial dysfunction observed in ARS, and its use might negatively impact the nasal mucosa causing issues such as dryness, stinging, burning, rebound congestion, as well as atrophy. In light of this, a novel nasal spray formulation containing both xylometazoline and hyaluronic acid (HA) was developed to provide a more effective and safer treatment for viral ARS. HA is a natural polysaccharide known to hydrate and moisturise the upper respiratory tract, maintain the integrity of the nasal mucosa, and promote mucociliary clearance and wound healing. To investigate the potential of this combination, this study was conducted using the nasal MucilAirTMin vitro model and high-speed phase-contrast microscopy to examine the effect of xylometazoline and HA on ciliary function by measuring ciliary beat frequency and their cytotoxicity by morphological, histological and ultrastructural analysis. This research is the first to assess the effects of a specific dose and molecular weight of HA as an active pharmaceutical ingredient in nasal spray formulations. The combination of a fast-acting decongestant and an additional active agent targeting nasal epithelial dysfunction has the potential to provide an improved, reliable and safe treatment for viral ARS, and may serve as the basis for future clinical studies.

Role-model, reoffer, reward: A thematic analysis and TDF mapping of influences on families' use of evidence-based vegetable feeding practices.

Children's vegetable intake is low, despite benefits for immediate and long-term health. Repeatedly reoffering vegetables, role-modelling consumption, and offering non-food rewards effectively increase children's vegetable acceptance and intake. However, a number of barriers prevent families from reoffering previously-rejected vegetables. This study used the Theoretical Domains Framework (TDF) and the COM-B model of behaviour to explore barriers and enablers to reoffering, role-modelling and offering non-food rewards among parents of 2-4-year-old children. Twenty-five semi-structured interviews were conducted, from which eleven core inductive themes were generated: 'Child factors', 'Eating beliefs', 'Effectiveness beliefs', 'Past experience', 'Current family behaviours', 'Harms', 'Knowledge', 'Need for change', 'Parent effort', 'Parent values' and 'Practical issues'. The codes underpinning these themes were inductively mapped to 11 of the 14 TDF domains, and five of the six COM-B components. Previously-reported influences on families' vegetable feeding practices were confirmed, including concerns about child rejection of foods/meals, cost of vegetables, and food waste. Novel findings included some parents' perceptions that these practices are pressurising, and that certain beliefs/knowledge about children's eating behaviour can provide a "protective mindset" that supports families' perseverance with reoffering over time. Future interventions should be tailored to better reflect the diversity of needs and previous experiences of feeding that families have, with some families likely to find that troubleshooting and further signposting is appropriate for their needs while others might benefit from more persuasive and educational approaches. The mapping of codes to the TDF and COM-B will facilitate the identification of appropriate intervention functions and behaviour change techniques when designing new interventions to support families with increasing their children's vegetable intake.

Four-week repeated exposure to tire-derived 6-PPD quinone causes multiple organ injury in male BALB/c mice.

N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine quinone (6-PPDQ) is the ozonation product of tire antioxidant 6-PPD. 6-PPDQ can be detected in different environments, such as roadway runoff and dust. Although 6-PPDQ toxicity has been frequently assessed in aquatic organisms, the possible toxic effects of 6-PPDQ on mammals remain largely unclear. We here aimed to perform systematic assessment to evaluate 6-PPDQ toxicity on multiple organs in mice. Male BALB/c mice were intraperitoneally injected with 6-PPDQ for two exposure modes, single intraperitoneal injection and repeated intraperitoneal injection every four days for 28 days. Serum, liver, kidney, lung, spleen, testis, brain, and heart were collected for injury evaluation by organ index, histopathology analysis and biochemical parameters. In 0.4 and 4 mg/kg 6-PPDQ single injected mice, no significant changes in organ indexes and biochemical parameters were detected, and only moderate pathological changes were observed in organs of liver, kidney, lung, and brain. Very different from this, in 0.4 and 4 mg/kg 6-PPDQ repeated injected mice, we observed the obvious increase in organ indexes of liver, kidney, lung, testis, and brain, and the decrease in spleen index. Meanwhile, the significant pathological changes were formed in liver, kidney, lung, spleen, testis, and brain in 0.4 and 4 mg/kg 6-PPDQ repeated injected mice. Biochemical parameters of liver (alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP)) and kidney (urea and creatinine) were all significantly upregulated by repeated injection with 0.4 and 4 mg/kg 6-PPDQ. After repeated exposure, most of 6-PPDQ was accumulated in liver and lung of mice. Therefore, our results suggested the risk of repeated exposure to 6-PPDQ in inducing toxicity on multiple organs in mice.

Preschool- and childcare center-based interventions to increase fruit and vegetable intake in preschool children in the United States: a systematic review of effectiveness and behavior change techniques.

BACKGROUND: Fruit and vegetable (FV) consumption in children in the United States (US) is very low. Adequate FV consumption is required for proper development during childhood, and dietary habits are established during preschool-age and tend to persist into adulthood. As most U.S. preschool-aged children attend childcare or preschool, this may be an opportune time and setting to conduct interventions to improve FV intake. These interventions should be based in theory and use behavior change techniques (BCTs) to explain mechanisms for expected change. To date, no published reviews have examined the effectiveness of childcare- or preschool-based FV interventions in preschoolers and their use of theoretical frameworks and BCTs. METHODS: This systematic review was completed adhering to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Inclusion criteria were randomized controlled trials (RCTs) published between 2012 and 2022 of interventions to improve diet or FV intake in preschoolers (aged 2-5 years) in childcare or preschool-settings. A search of four databases was conducted between in September 2022 using search terms pertaining to the study's primary aim (FV consumption), age group (preschool-aged), settings (US childcare or preschool settings), and study design (RCT). Additional criteria were objective measures of FV consumption or skin carotenoids, as a proxy for FV intake. Included studies were narratively synthesized based on intervention type, measured effect, and use of theory and BCTs. RESULTS: The search resulted in six studies that reported on nine interventions. Overall, six interventions increased FV intake, of which five used nutrition education and one manipulated the feeding environment. Among the three interventions with no measured effect, two manipulated the feeding environment and one used peer modeling. Effective studies used at least three BCTs, though no pattern was observed between use of theory or BCTs and intervention effect. CONCLUSIONS: While several studies have shown promising results, the limited number of studies identified in this review highlights key gaps in this field: there is a need for studies to test FV interventions in US childcare settings that use objective measures of FV intake, directly compare intervention components and BCTs, are theory-based, and assess long-term behavior change.

A randomized, controlled trial of a Nordic, protein-reduced complementary diet in infants: effects on body composition, growth, biomarkers, and dietary intake at 12 and 18 months.

BACKGROUND: High intake of protein and low intake of plant-based foods during complementary feeding can contribute to negative long-term health effects. OBJECTIVES: To investigate the effects of a protein-reduced, Nordic complementary diet on body composition, growth, biomarkers, and dietary intake, compared with current Swedish dietary recommendations for infants at 12 and 18 mo. METHODS: Healthy, term infants (n = 250) were randomly allocated to either a Nordic group (NG) or a conventional group (CG). From 4 to 6 mo, NG participants received repeated exposures of Nordic taste portions. From 6 to 18 mo, NG was supplied with Nordic homemade baby food recipes, protein-reduced baby food products, and parental support. CG followed the current Swedish dietary recommendations. Measurements of body composition, anthropometry, biomarkers, and dietary intake were collected from baseline and at 12 and 18 mo. RESULTS: Of the 250 infants, 82% (n = 206) completed the study. There were no group differences in body composition or growth. In NG, protein intake, blood urea nitrogen and plasma IGF-1 were lower compared to CG at 12 and 18 mo. Infants in NG consumed 42% to 45% more fruits and vegetables compared to CG at 12 and 18 mo, which was reflected in a higher plasma folate at 12 and 18 mo. There were no between-group differences in EI or iron status. CONCLUSIONS: Introduction of a predominantly plant-based, protein-reduced diet as part of complementary feeding is feasible and can increase fruit and vegetable intake. This trial was registered at clinicaltrials.gov as NCT02634749.

Promoting vegetable acceptance in toddlers using a contingency management program: A cluster randomised trial.

Children learn to accept novel foods by repeated exposure to these foods. In the current study, we investigated in toddlers whether a contingency management program (The Vegetable Box), comprising repeated vegetable taste exposure with contingent non-food rewards, is particularly potent at increasing recognition of and willingness to try vegetables. A total of 598 children (1-4 years old) recruited at 26 different day-care centres in the Netherlands took part. The day-care centres were randomly assigned to one of three conditions ('exposure/reward', 'exposure/no reward', or 'no exposure/no reward'). At the start and directly after the 3-month intervention period, all children were asked to identify various vegetables (recognition test; max score = 14) and whether they would like to taste and consume 1 or 2 bite-size pieces of tomato, cucumber, carrot, bell pepper, radish, and cauliflower (willingness to try test). Data were analysed with linear mixed-effects regression analyses (for recognition and willingness to try separately) with condition and time as independent variables, and adjusting for day-care centre clustering. Vegetable recognition significantly increased in both the 'exposure/reward' and the 'exposure/no reward' group, relative to the 'no exposure/no reward' control group. The willingness to try vegetables only increased significantly in the 'exposure/reward' group. Offering vegetables to children at day-care centres significantly increased toddlers' ability to identify various vegetables, but rewards contingent upon tasting vegetables appear particularly effective to also increase children's willingness to try (i.e., taste and consume) different vegetables. This result corroborates and strengthens previous findings demonstrating the efficacy of similar reward-based programs.