Development of a tool for Individual Aquatic Risk Management among children of 6-12 years (IARM-C).

Background: When it comes to children’s competence in, on and around water, their risk management also plays an important role. For an optimal and save participation in aquatic recreation, there is a crucial need to communicate about a realistic perception of potential dangers in relation to one’s own actual and perceived aquatic skills. Goals: The aim of this study was to develop and validate a tool for Individual Aquatic Risk Management for Children (IARM-C) useful in both research and practice regarding water safety for elementary school children and their families, as offered in schools, by local communities and different (water) sport organisations. Method: The IARM-C tool was developed and validated in three subsequent phases: (1) a selection of relevant aquatic situations with possible risks for children based on the literature and discussed with experts, resulting in 10 aquatic situations that were drawn, (2) a pilot study with 22 children to test content (face) validity, and (3) a cross-sectional study with 70 children (6-12 years, 35 girls and 35 boys, 8.9 ± 2.0 years) recruited via convenience sampling in different (swimming) schools in Brussel (Belgium) to test their risk perception, assessment and decision making in these 10 situations. Results: For each of the 10 aquatic risk situations of the IARM-C, data collection was organised in a one-on-one interview in order to assist the child in completing the questionnaire. Six of the 10 pictures resulted in a correct risk perception for > 80% (range between 83-94%) of the children. For one drawn aquatic risk situation in the swimming pool context (i.e. falling on someone else), only 60% of the children gave a correct description of the situation. In the drawn open water aquatic risk situations, three pictures scored quite low (range between 49%-54%): warning flag at sea, dangerous objects and sandbank in the sea. Conclusions: The IARM-C tool, showing pictures of aquatic risk situations followed by three categories of questions (risk perception, assessment and decision making), is a useful instrument for further research and education purposes, especially for the swimming pool cases.Desarrollo de una herramienta para la Gestión individual del riesgo acuático entre los niños de 6 a 12 años (IARM-C) Resumen Antecedentes: Cuando se trata de la competencia de los niños en, sobre y alrededor del agua, su competencia de riesgo también juega un papel importante. Para una participación óptima y segura en la recreación acuática, existe una necesidad crucial de comunicar una percepción realista de los peligros potenciales en relación con las propias habilidades acuáticas reales y percibidas. Objetivos: El objetivo de este estudio fue desarrollar y validar una herramienta para la Gestión Individual de Riesgos Acuáticos para Niños (IARM-C) útil tanto en la investigación como en la práctica en relación con la seguridad en el agua para los niños de la escuela primaria y sus familias, tal y como se ofrece en las escuelas, por las comunidades locales y diferentes organizaciones deportivas (acuáticas). Método: La herramienta IARM-C se desarrolló y validó en tres fases sucesivos: (1) una selección de situaciones acuáticas relevantes con posibles riesgos para los niños, basada en la literatura y discutida con expertos, lo que dio como resultado 10 ilustraciones de situaciones acuáticos, (2) un estudio piloto con 22 niños para comprobar la validez (aparente) del contenido, y (3) un estudio transversal con 70 niños (35 niñas y 35 niños, 6-12 años, 8.9 ± 2.0 años) seleccionados mediante muestreo de conveniencia en diferentes escuelas (de natación) de Bruselas (Bélgica) para comprobar su percepción de los riesgos, su evaluación y su toma de decisiones en estas 10 situaciones. Resultados: Para cada una de las 10 situaciones de riesgo acuático del IARM-C, la recogida de datos se organizó en una entrevista individual para ayudar al niño a rellenar el cuestionario. Seis de los 10 dibujos dieron lugar a una percepción de riesgo correcta para más del 80% (rango entre 83-94%) de los niños. En una de las situaciones de riesgo acuático dibujadas en el contexto de la piscina (es decir, la caída sobre otra persona), sólo el 60% de los niños dio una descripción correcta de la situación. En las situaciones de riesgo acuático dibujadas en aguas abiertas, tres imágenes obtuvieron una puntuación bastante baja (entre el 49% y el 54%): bandera de advertencia en el mar, objetos peligrosos y banco de arena en el mar. Conclusiones: La herramienta IARM-C, que muestra imágenes de situaciones de riesgo acuático seguidas de tres categorías de preguntas (percepción del riesgo, evaluación y toma de decisiones), es un instrumento útil para seguir investigando y educando, especialmente para los casos de piscinas

GlycoVHH : optimal sites for introducing N-glycans on the camelid VHH antibody scaffold and use for macrophage delivery

As small and stable high-affinity antigen binders, VHHs boast attractive characteristics both for therapeutic use in various disease indications, and as versatile reagents in research and diagnostics. To further increase the versatility of VHHs, we explored the VHH scaffold in a structure-guided approach to select regions where the introduction of an N-glycosylation N-X-T sequon and its associated glycan should not interfere with protein folding or epitope recognition. We expressed variants of such glycoengineered VHHs in the Pichia pastoris GlycoSwitchM5 strain, allowing us to pinpoint preferred sites at which Man(5)GlcNAc(2)-glycans can be introduced at high site occupancy without affecting antigen binding. A VHH carrying predominantly a Man(5)GlcNAc(2) N-glycan at one of these preferred sites showed highly efficient, glycan-dependent uptake by Mf4/4 macrophages in vitro and by alveolar lung macrophages in vivo, illustrating one potential application of glyco-engineered VHHs: a glycan-based targeting approach for lung macrophage endolysosomal system delivery. The set of optimal artificial VHH N-glycosylation sites identified in this study can serve as a blueprint for targeted glyco-engineering of other VHHs, enabling site-specific functionalization through the rapidly expanding toolbox of synthetic glycobiology

An affinity-enhanced, broadly neutralizing heavy chain-only antibody protects against SARS-CoV-2 infection in animal models

Broadly neutralizing antibodies are an important treatment for individuals with coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Antibody-based therapeutics are also essential for pandemic preparedness against future Sarbecovirus outbreaks. Camelid-derived single domain antibodies (VHHs) exhibit potent antimicrobial activity and are being developed as SARS-CoV-2-neutralizing antibody-like therapeutics. Here, we identified VHHs that neutralize both SARS-CoV-1 and SARS-CoV-2, including now circulating variants. We observed that the VHHs bound to a highly conserved epitope in the receptor binding domain of the viral spike protein that is difficult to access for human antibodies. Structure-guided molecular modeling, combined with rapid yeast-based prototyping, resulted in an affinity enhanced VHH-human immunoglobulin G1 Fc fusion molecule with subnanomolar neutralizing activity. This VHH-Fc fusion protein, produced in and purified from cultured Chinese hamster ovary cells, controlled SARS-CoV-2 replication in prophylactic and therapeutic settings in mice expressing human angiotensin converting enzyme 2 and in hamsters infected with SARS-CoV-2. These data led to affinity-enhanced selection of the VHH, XVR011, a stable anti-COVID-19 biologic that is now being evaluated in the clinic