Analysis of nanomaterial biocoronas in biological and environmental surroundings.

A biomolecular coating, or biocorona, forms on the surface of engineered nanomaterials (ENMs) immediately as they enter biological or environmental systems, defining their biological and environmental identity and influencing their fate and performance. This biomolecular layer includes proteins (the protein corona) and other biomolecules, such as nucleic acids and metabolites. To ensure a meaningful and reproducible analysis of the ENMs-associated biocorona, it is essential to streamline procedures for its preparation, separation, identification and characterization, so that studies in different labs can be easily compared, and the information collected can be used to predict the composition, dynamics and properties of biocoronas acquired by other ENMs. Most studies focus on the protein corona as proteins are easier to monitor and characterize than other biomolecules and play crucial roles in receptor engagement and signaling; however, metabolites play equally critical roles in signaling. Here we describe how to reproducibly prepare and characterize biomolecule-coated ENMs, noting especially the steps that need optimization for different types of ENMs. The structure and composition of the biocoronas are characterized using general methods (transmission electron microscopy, dynamic light scattering, capillary electrophoresis-mass spectrometry and liquid chromatography-mass spectrometry) as well as advanced techniques, such as transmission electron cryomicroscopy, synchrotron-based X-ray absorption near edge structure and circular dichroism. We also discuss how to use molecular dynamic simulation to study and predict the interaction between ENMs and biomolecules and the resulting biocorona composition. The application of this protocol can provide mechanistic insights into the formation, composition and evolution of the ENM biocorona, ultimately facilitating the biomedical and agricultural application of ENMs and a better understanding of their impact in the environment.

A template wizard for the cocreation of machine-readable data-reporting to harmonize the evaluation of (nano)materials.

Making research data findable, accessible, interoperable and reusable (FAIR) is typically hampered by a lack of skills in technical aspects of data management by data generators and a lack of resources. We developed a Template Wizard for researchers to easily create templates suitable for consistently capturing data and metadata from their experiments. The templates are easy to use and enable the compilation of machine-readable metadata to accompany data generation and align them to existing community standards and databases, such as eNanoMapper, streamlining the adoption of the FAIR principles. These templates are citable objects and are available as online tools. The Template Wizard is designed to be user friendly and facilitates using and reusing existing templates for new projects or project extensions. The wizard is accompanied by an online template validator, which allows self-evaluation of the template (to ensure mapping to the data schema and machine readability of the captured data) and transformation by an open-source parser into machine-readable formats, compliant with the FAIR principles. The templates are based on extensive collective experience in nanosafety data collection and include over 60 harmonized data entry templates for physicochemical characterization and hazard assessment (cell viability, genotoxicity, environmental organism dose-response tests, omics), as well as exposure and release studies. The templates are generalizable across fields and have already been extended and adapted for microplastics and advanced materials research. The harmonized templates improve the reliability of interlaboratory comparisons, data reuse and meta-analyses and can facilitate the safety evaluation and regulation process for (nano) materials.

Daphnia as a model organism to probe biological responses to nanomaterials-from individual to population effects via adverse outcome pathways.

The importance of the cladoceran Daphnia as a model organism for ecotoxicity testing has been well-established since the 1980s. Daphnia have been increasingly used in standardised testing of chemicals as they are well characterised and show sensitivity to pollutants, making them an essential indicator species for environmental stress. The mapping of the genomes of D. pulex in 2012 and D. magna in 2017 further consolidated their utility for ecotoxicity testing, including demonstrating the responsiveness of the Daphnia genome to environmental stressors. The short lifecycle and parthenogenetic reproduction make Daphnia useful for assessment of developmental toxicity and adaption to stress. The emergence of nanomaterials (NMs) and their safety assessment has introduced some challenges to the use of standard toxicity tests which were developed for soluble chemicals. NMs have enormous reactive surface areas resulting in dynamic interactions with dissolved organic carbon, proteins and other biomolecules in their surroundings leading to a myriad of physical, chemical, biological, and macromolecular transformations of the NMs and thus changes in their bioavailability to, and impacts on, daphnids. However, NM safety assessments are also driving innovations in our approaches to toxicity testing, for both chemicals and other emerging contaminants such as microplastics (MPs). These advances include establishing more realistic environmental exposures via medium composition tuning including pre-conditioning by the organisms to provide relevant biomolecules as background, development of microfluidics approaches to mimic environmental flow conditions typical in streams, utilisation of field daphnids cultured in the lab to assess adaption and impacts of pre-exposure to pollution gradients, and of course development of mechanistic insights to connect the first encounter with NMs or MPs to an adverse outcome, via the key events in an adverse outcome pathway. Insights into these developments are presented below to inspire further advances and utilisation of these important organisms as part of an overall environmental risk assessment of NMs and MPs impacts, including in mixture exposure scenarios.

Alaska's Play Every Day Campaign Encourages Parents to Serve Healthy Drinks to Young Children.

BACKGROUND: Sugary drink consumption by young children is a public health concern. The State of Alaska, partnering with the Alaska Native Tribal Health Consortium, implemented the Play Every Day social marketing campaign in 2019-2021 to encourage parents to serve healthy drinks to young children. The campaign's intended audience was parents who experience disproportionately poor nutrition outcomes: Alaska Native people, those living in rural communities, and those with low incomes and/or educational attainment. We described campaign development, implementation, and performance. METHOD: Parents from the identified disproportionately affected populations participated in formative research. Campaign awareness and engagement questions were added to Alaska's child health surveillance system. Regression models assessed associations between campaign exposure and outcomes. RESULTS: The sample included 476 Alaska mothers of 3-year-old children. Of the 34% who reported seeing the campaign, 21% said they changed drinks served to their child because of the campaign. Campaign awareness, engagement, and reported changes in drinks given to children because of the campaign were greater among Alaska Native mothers than White mothers. Among all mothers, those who said the campaign gave them new information or that they shared the campaign had 7 to 8 times greater odds for reporting changes in behavior than those not engaged with the campaign. IMPLICATIONS FOR PRACTICE: Social marketing campaigns that encourage parents to serve healthy drinks to young children may change behavior. Resources should be systematically dedicated to research, implementation, and evaluation focused on specific populations. Partnering with trusted community-serving organizations likely improves outcomes in disproportionately affected populations.

Seabird stress and breeding: Endocrine and hematological stress biomarkers differ between gray-faced petrel (Pterodroma gouldi) colonies.

Seabird breeding success is known to reflect oceanic conditions. Gray-faced petrels (Pterodroma gouldi) breeding on the east coast of Auckland, New Zealand, exhibit poor reproductive success and slow chick development compared to west coast conspecifics. This study mapped changes in physiological traits (corticosterone [CORT] and hematological parameters) indicative of sublethal stress in this Procellariiform species between the west coast (Ihumoana) and east coast (Hawere) island colonies. We found adult gray-faced petrels on the east coast to be lighter and, unlike west coast birds, exhibited an attenuation of response CORT levels between incubation and chick-rearing phases. Such responses were also reflected in east coast chicks that were lighter and had higher feather CORT titers than west coast chicks. Measures of adult hematology and plasma biochemistry revealed significantly lower glucose levels in east coast birds and indicated that chick rearing is the most stressful phase of breeding for this species Combined; these results suggest that east coast birds are under greater nutritional stress and that parents appear to transfer the costs of poor foraging to their chicks to preserve their own condition, consequently increasing chick developmental stress. Our results suggest that any long-term decrease in ocean conditions and/or climatic shifts would be more acutely felt by east coast chicks and potentially their parents, resulting in years of poor breeding success rates on a local scale.

Blood tests are not always helpful in predicting bacterial meningitis in children.

Bacterial meningitis is characterized by inflammation of the meninges covering the brain. It is a life-threatening illness, if untreated. The aim of this study was to examine whether blood tests including C-reactive protein (CRP) and full blood count (FBC) predict bacterial meningitis in children. We also examined the relationship between cerebrospinal fluid (CSF) tests including gram stain, culture and polymerase chain reaction (PCR) and blood tests such as culture and PCR results. We studied 11 patients admitted with bacterial meningitis in the Paediatric wards, National Children's Hospital, Dublin, Northern Ireland (2012-2016). The mean age was 10.7 (SD, 14.7) months. In this group of patients, 5 (45.5%) had leucocytosis, 7 (63.6%) experienced neutrophilia and 1 (9.1%) had both leukopenia and neutropenia. C-reactive protein (CRP) value of less than 1 mg/L was found in 2 patients (18.2%). No link was found between CSF gram stain and CSF culture (p value 0.66) or CSF PCR results (p value 0.75). Meningitis should be investigated and treated if clinically suspected, regardless of CRP values or peripheral blood results.

Process and outcomes of patient-centered medical care with Alaska Native people at Southcentral Foundation.

PURPOSE: This study describes key elements of the transition to a patient-centered medical home (PCMH) model at Southcentral Foundation (SCF), a tribally owned and managed primary care system, and evaluates changes in emergency care use for any reason, for asthma, and for unintentional injuries, during and after the transition. METHODS: We conducted a time series analyses of emergency care use from medical record data. We also conducted 45 individual, in-depth interviews with PCMH patients (customer-owners), primary care clinicians, health system employees, and tribal leaders. RESULTS: Emergency care use for all causes was increasing before the PCMH implementation, dropped during and immediately after the implementation, and subsequently leveled off. Emergency care use for adult asthma dropped before, during, and immediately after implementation, subsequently leveling off approximately 5 years after implementation. Emergency care use for unintentional injuries, a comparison variable, showed an increasing trend before and during implementation and decreasing trends after implementation. Interview participants observed improved access to primary care services after the transition to the PCMH tempered by increased staff fatigue. Additional themes of PCMH transformation included the building of relationships for coordinated, team-based care, and the important role of leadership in PCMH implementation. CONCLUSIONS: All reported measures of emergency care use show a decreasing trend after the PCMH implementation. Before the implementation, overall use and use for unintentional injuries had been increasing. The combined quantitative and qualitative results are consistent with decreased emergency care use resulting from a decreased need for emergency care services due to increased availability of primary care services and same-day appointments.

Comparison of p16INK4A and Hybrid Capture 2 human papillomavirus testing as adjunctive tests in liquid-based gynecologic SurePath preparations.

p16(INK4a), cyclin-dependent kinase inhibitor, is functionally inactivated in many tumors, including cervical cancer. We compared p16(INK4A) immunocytochemical staining and Hybrid Capture 2 (HCII) on SurePath specimens using tissue biopsies (as the gold standard). Their utility in a spectrum of atypical and preneoplastic lesions, and their ability to accurately identify underlying lesions of CIN II or greater was assessed using biopsy follow-up data. One-hundred and seventeen residual SurePath samples were collected: 43 atypical squamous cells of undetermined significance (ASCUS), 47 low-grade (LGSIL), and 27 high-grade (HGSIL) squamous intraepithelial lesions. Two slides were prepared from each sample; one stained with the SurePath autocyte stain and one immunostained using the CINtec p16(INK4a) Cytology Kit (Dakocytomation). High-risk HPV testing was performed using the HCII DNA test (Digene, Gaithersburg, MD). Available tissue biopsy follow-up data was retrieved. p16(INK4a) was positive in 32.6% (14/43) ASCUS, 46.8% (22/47) LGSIL, and 48.1% (13/27) HGSIL specimens. HCII DNA test was positive in 41.9% (18/43) ASCUS, 78.7% (37/47) LGSIL, and 96.3% (26/27) HGSIL samples. The sensitivity, specificity, positive (PPV) and negative (NPV) predictive values of p16(INK4a) and HCII were: 58.7% and 89.8%, 58.6% and 34.6%, 69.2% and 72.1%, 47.2% and 64.3%, respectively. In patients with cervical biopsies, the PPV of HCII (92.3%) results for a biopsy with CINII/III was significantly higher than the PPV of p16(INK4a) (52%) (P=0.001). Using liquid-based cytology specimens, HCII is a more sensitive test than p16(INK4a) for detection of abnormal cytology. HCII has a higher PPV than p16(INK4a) for identifying CIN II/III.