RESUMEN
The inability of visible light to penetrate far through biological tissue limits its use for phototherapy and photodiagnosis of deep-tissue sites of disease. This is unfortunate because many visible dyes are excellent photosensitizers and photocatalysts that can induce a wide range of photochemical processes, including photogeneration of reactive oxygen species. One potential solution is to bring the light source closer to the site of disease by using a miniature implantable LED. With this goal in mind, we fabricated a wireless LED-based device (volume of 23 mm3) that is powered by RF energy and emits light with a wavelength of 573 nm. It has the capacity to excite the green absorbing dye Rose Bengal, which is an efficient type II photosensitizer. The wireless transfer of RF power is effective even when the device is buried in chicken breast and located 6 cm from the transmitting antenna. The combination of a wireless device as light source and Rose Bengal as photosensitizer was found to induce cell death of cultured HT-29 human colorectal adenocarcinoma cells. Time-dependent generation of protruding bubbles was observed in the photoactivated cells suggesting cell death by light-induced pyroptosis and supporting evidence was gained by cell staining with the fluorescence probes Annexin-V FITC and Propidium Iodide. The results reveal a future path towards a wireless implanted LED-based device that can trigger photodynamic immunogenic cell death in deep-seated cancerous tissue.
Asunto(s)
Fotoquimioterapia , Fármacos Fotosensibilizantes , Piroptosis , Rosa Bengala , Fármacos Fotosensibilizantes/farmacología , Piroptosis/efectos de los fármacos , Fotoquimioterapia/métodos , Humanos , Rosa Bengala/farmacología , Células HT29 , Tecnología Inalámbrica , AnimalesRESUMEN
In response to the SARS-CoV-2 pandemic, we developed a multiplexed, paired-pool droplet digital PCR (MP4) screening assay. Key features of our assay are the use of minimally processed saliva, 8-sample paired pools, and reverse-transcription droplet digital PCR (RT-ddPCR) targeting the SARS-CoV-2 nucleocapsid gene. The limit of detection was determined to be 2 and 12 copies per µl for individual and pooled samples, respectively. Using the MP4 assay, we routinely processed over 1,000 samples a day with a 24-h turnaround time and over the course of 17 months, screened over 250,000 saliva samples. Modeling studies showed that the efficiency of 8-sample pools was reduced with increased viral prevalence and that this could be mitigated by using 4-sample pools. We also present a strategy for, and modeling data supporting, the creation of a third paired pool as an additional strategy to employ under high viral prevalence.
Asunto(s)
COVID-19 , SARS-CoV-2 , Humanos , SARS-CoV-2/genética , Saliva/química , ARN Viral/genética , Reacción en Cadena de la Polimerasa , Sensibilidad y Especificidad , Prueba de COVID-19RESUMEN
Nested hierarchical structure is one of life's most familiar properties and a major component of biological diversity and complexity. However, there is little effort to teach the evolution of the hierarchy of life, as there is little effort to teach biological complexity per se. We propose a framework for teaching biological complexity based on research on evolutionary transitions in individuality (ETI theory). Translating ETI theory into the classroom allows students to see the connections between natural selection, social behavior in groups, and the major landmarks of biodiversity in the hierarchy of life. The translation of ETI theory into pedagogic content and practices involves (i) the new content that must be taught, (ii) the development of general teaching tools to teach this new content, and (iii) connecting the new content and teaching tools to the specific educational context including integrating with learning standards and benchmarks. We show how teaching ETIs aids in the teaching of science practices and in teaching the process of evolutionary change. Evolutionary transitions research provides a way to teach biological complexity that is familiar and engaging to students, leveraging their inherent understanding of social dynamics and group behavior.