Use of RNAlater in fluorescence-activated cell sorting (FACS) reduces the fluorescence from GFP but not from DsRed

<p>Abstract</p> <p>Background</p> <p>Flow cytometry utilizes signals from fluorescent markers to separate targeted cell populations for gene expression studies. However, the stress of the FACS process could change normal gene expression profiles. RNAlater could be used to stop such changes in original gene expression profiles through its ability to denature RNase and other proteins. The normal conformational structure of fluorescent proteins must be maintained in order to fluoresce. Whether or not RNAlater would affect signals from different types of intrinsic fluorescent proteins is crucial to its use in flow cytometry; this question has not been investigated in detail.</p> <p>Findings</p> <p>To address this question, we analyzed the effect of RNAlater on fluorescence intensity of GFP, YFP, DsRed and small fluorescent molecules attached to secondary antibodies (Cy2 and Texas-Red) when used in flow cytometry. FACS results were confirmed with fluorescence microscopy. Our results showed that exposure of YFP and GFP containing cells to RNAlater reduces the intensity of their fluorescence to such an extent that separation of such labeled cells is difficult if not impossible. In contrast, signals from DsRed2, Cy2 and Texas-Red were not affected by RNAlater treatment. In addition, the background fluorescence and clumping of dissociated cells are altered by RNAlater treatment.</p> <p>Conclusions</p> <p>When considering gene expression studies using cell sorting with RNAlater, DsRed is the fluorescent protein of choice while GFP/YFP have severe limitations because of their reduced fluorescence. It is necessary to examine the effects of RNAlater on signals from fluorescent markers and the physical properties (e.g., clumping) of the cells before considering its use in cell sorting.</p

Responding to Food Insecurity and Community Crises through Food Policy Council Partnerships in a Rural Setting

Food insecurity is a constant struggle for many communities and food needs are often amplified during times of crisis. The objective of this article is to describe the progress in our community food policy council partnership by presenting the elements of success in facilitating policy change and programs that have helped our community in rural Pennsylvania respond to a variety of challenges, including the coronavirus disease 2019 (COVID-19) crisis. We also critically examine ongoing challenges and implications for our work. The elements that have contributed to sustaining our collective work include having a common agenda; collaboration; maintaining independent but mutually supporting member organizational goals; valuing those who are most impacted; and continuous communication. By applying these elements of partnership, the council remains focused on healthy food access, particularly during this COVID-19 crisis and ongoing food-related inequities

Voices of the Street: The Din from Within

Content Warning:&nbsp;The stories in this series deal with difficult and sometimes traumatic topics. Please practice self care, stop listening, and seek help if you need to. Scroll down to find links to available supports. &nbsp; Be transported into the soundscapes of two different poems. This installment of the Voices of the Street podcast is produced by Jules Chapman, a writer and peer support worker who is deeply involved in the Downtown Eastside community.&nbsp; Jules reads from the Voices of the Street anthology, sharing Elaine Schell’s “The Din from Within” and the original poem that Jules was inspired to write in response. These pieces take us through writerly reflections on poetic expression, getting to know yourself, processing through writing and sharing creative work with the world

Management of patients with rare adult solid cancers: objectives and evaluation of European reference networks (ERN) EURACAN

Summary: About 500,000 patients with rare adult solid cancers (RASC) are diagnosed yearly in Europe. Delays and unequal quality of management impact negatively their survival. Since 2017, European reference networks (ERN) aim to improve the quality of care of patients with rare disease. The steering committee of EURACAN, including physicians, researchers and patients review here the previous actions, present objectives of the ERN EURACAN dedicated to RASC. EURACAN promoted management in reference centres, and equal implementation of excellence and innovation in Europe and developed 22 clinical practice guidelines (CPGs). Additionally, fourteen information brochures translated in 24 EU languages were developed in collaboration with patient advocacy groups (ePAGs) and seventeen training session were organized. Nevertheless, connections to national networks in the 26 participating countries (106 centres), simplification of cross-border healthcare, international multidisciplinary tumour boards, registries and monitoring of the quality of care are still required. In this Health Policy, evaluation criteria of the performances of the network and of health care providers are proposed