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Exosomes and other extracellular vesicles (commonly referred to as EVs) have generated a lot of attention for their potential applications in both diagnostics and therapeutics. The contents of these vesicles are the subject of intense research, and the relatively recent discovery of RNA inside EVs has raised interest in the biological function of these RNAs as well as their potential as biomarkers for cancer and other diseases. Traditional ultracentrifugation-based protocols to isolate EVs are labor-intensive and subject to significant variability. Various attempts to develop methods with robust, reproducible performance have not yet been completely successful. Here, we report the development and characterization of a spin column-based method for the isolation of total RNA from EVs in serum and plasma. This method isolates highly pure RNA of equal or higher quantity compared to ultracentrifugation, with high specificity for vesicular over non-vesicular RNA. The spin columns have a capacity to handle up to 4 mL sample volume, enabling detection of low-abundance transcripts in serum and plasma. We conclude that the method is an improvement over traditional methods in providing a faster, more standardized way to achieve reliable high quality RNA preparations from EVs in biofluids such as serum and plasma. The first kit utilizing this new method has recently been made available by Qiagen as \"exoRNeasy Serum/Plasma Maxi Kit\".

Background: Liquid biopsies often aim to compare the amount of nucleic acids of tumour origin with nucleic acids that are shed from healthy cells. While it is understood that the amount of tumour material in circulation strongly depends on tumour stage, the conditions influencing the amount of wildtype nucleic acids are less well described. The amount of RNA contained in exosomes and other extracellular vesicles (exoRNA), as well as the amount of cell-free DNA (cfDNA), may be influenced by preanalytical parameters - potentially changing the interpretation of a liquid biopsy result. Here, we analyse the release of exoRNA and cfDNA by physical activity. Methods: We sampled the blood of four healthy individuals by venipuncture before and after 30 min of continuous physical exercise and used a combined co-extraction of exoRNA and cfDNA to analyse their relative abundance in plasma. Size distribution of the total nucleic acids was accessed using a BioAnalyzer 6000 Pico Kit. The content of exoRNA was quantified using RT-qPCR for specific mRNAs (GAPDH and RPL0) and the content of cfDNA was quantified using primers for the loci of often used oncogenes (KRAS, BRAF and PIK3CA). Results: All four individuals showed increased amount of total nucleic acids after physical exercise, mainly corresponding to the size of monoand dinucleosomal DNA. Comparing the pre- and post-exercise datapoints of each patient, the increase in cfDNA was measured consistently between 10 and 30-fold, while the increase in exosomal mRNAs was only two-fold on average. Summary/Conclusion: While both exoRNA and cfDNA increased already after 30 min of exercise, the increase in cfDNA consistently exceeded the increase of exosomal RNA by an order of magnitude. This effect of physical activity has to be taken into account when interpreting datasets that use the absolute or relative amount of nucleic acids in liquid biopsies.

Exosomes and other extracellular vesicles (commonly referred to as EVs) have generated a lot of attention for their potential applications in both diagnostics and therapeutics. The contents of these vesicles are the subject of intense research, and the relatively recent discovery of RNA inside EVs has raised interest in the biological function of these RNAs as well as their potential as biomarkers for cancer and other diseases. Traditional ultracentrifugation-based protocols to isolate EVs are labor-intensive and subject to significant variability. Various attempts to develop methods with robust, reproducible performance have not yet been completely successful. Here, we report the development and characterization of a spin column-based method for the isolation of total RNA from EVs in serum and plasma. This method isolates highly pure RNA of equal or higher quantity compared to ultracentrifugation, with high specificity for vesicular over non-vesicular RNA. The spin columns have a capacity to handle up to 4 mL sample volume, enabling detection of low-abundance transcripts in serum and plasma. We conclude that the method is an improvement over traditional methods in providing a faster, more standardized way to achieve reliable high quality RNA preparations from EVs in biofluids such as serum and plasma. The first kit utilizing this new method has recently been made available by Qiagen as \"exoRNeasy Serum/Plasma Maxi Kit\".

Background Eating disorders are increasing in prevalence and confer serious physical, emotional and social impacts on individuals, families, communities and systems. Tertiary education of health professionals is key to addressing these impacts. Yet, graduates of key health professions may not complete their studies with the necessary knowledge, skills and confidence to prevent, identify, refer and provide safe care within their role and scope of practice. This study reports on current eating disorder-focused teaching, and opportunities for improvements, within Australian tertiary training programs for key medical and allied health professional groups. Methods Tertiary leaders and educators ( n  = 205) representing 42 Australian tertiary education institutions (universities and private education organizations) completed an online survey including questions that explored the relevance, importance, barriers and enablers of eating disorder-focused teaching. Results Three quarters (76.1%) of participants indicated that students had the opportunity to learn about eating disorders within the current curriculum. Identification (54.1%) and treatment (46.3%) were most likely to be taught; prevention (31.2%) and psychological support (38.0%) were less likely to be taught. Most respondents (85.9%) reported that teaching about eating disorders, disordered eating and body image within their program was either important (23.9%) or very important (62.0%). Perceived challenges to enhancing content included a crowded curriculum, lack of appropriately skilled educators and lack of clarity regarding basic profession-specific competencies and when these should be taught. Potential facilitators include developing materials and networks for implementing best practice training, support of accreditation/professional bodies and use of lived experience educators. Conclusions Most Australian healthcare educators consider eating disorders important and relevant to their curricula. However, there are limited time and practical opportunities for learning about eating disorders within current curricula. There is a need for guidance and resource development to help support educators to enhance eating disorder content in relevant curricula. Plain Language Summary Of the 205 health professional educators that responded to the survey, most but not all provide an opportunity for students to learn about eating disorders, disordered eating and body image. Eating disorders are seen as an important clinical teaching area, particularly because of the severity, impact and increasing prevalence. However, barriers to including eating disorder content included competing with an already full curriculum and difficulty providing practical learning. Despite these barriers, educators described the need for contemporary, best practice teaching materials and a variety of learning opportunities aligned with discipline specific frameworks and scope would help support educators to provide appropriate, evidenced-based eating disorder content to students.

Point-of-care ultrasounds (US) are used during cardiopulmonary resuscitation (CPR) and after return of spontaneous circulation (ROSC). Carotid ultrasounds are a potential non-invasive monitoring tool for chest compressions, but their general value and feasibility during CPR are not fully determined. In this prospective observational study, we performed carotid US during conventional- and extracorporeal CPR and after ROSC with at least one transverse and coronal image, corresponding loops with and without color doppler, and pulsed-wave doppler loops. The feasibility of carotid US during (peri-)arrest and type and frequency of diagnostic findings were examined. We recruited 16 patients and recorded utilizable US images in 14 cases (88%; complete imaging protocols in 11 patients [69%]). In three of all patients (19%) and in 60% (3/5) of cases during CPR plus a full imaging protocol, we observed: (i) in one patient a collapse of the common carotid artery linked to hypovolemia, and (ii) in two patients a biphasic flow during CPR linked to prolonged low-flow time prior to admission and adverse outcome. Carotid artery morphology and carotid blood flow characteristics may serve as therapeutic target and prognostic parameters. However, future studies with larger sample sizes are needed.

Eating disorders are increasing in prevalence and confer serious physical, emotional and social impacts on individuals, families, communities and systems. Tertiary education of health professionals is key to addressing these impacts. Yet, graduates of key health professions may not complete their studies with the necessary knowledge, skills and confidence to prevent, identify, refer and provide safe care within their role and scope of practice. This study reports on current eating disorder-focused teaching, and opportunities for improvements, within Australian tertiary training programs for key medical and allied health professional groups. Tertiary leaders and educators (n = 205) representing 42 Australian tertiary education institutions (universities and private education organizations) completed an online survey including questions that explored the relevance, importance, barriers and enablers of eating disorder-focused teaching. Three quarters (76.1%) of participants indicated that students had the opportunity to learn about eating disorders within the current curriculum. Identification (54.1%) and treatment (46.3%) were most likely to be taught; prevention (31.2%) and psychological support (38.0%) were less likely to be taught. Most respondents (85.9%) reported that teaching about eating disorders, disordered eating and body image within their program was either important (23.9%) or very important (62.0%). Perceived challenges to enhancing content included a crowded curriculum, lack of appropriately skilled educators and lack of clarity regarding basic profession-specific competencies and when these should be taught. Potential facilitators include developing materials and networks for implementing best practice training, support of accreditation/professional bodies and use of lived experience educators. Most Australian healthcare educators consider eating disorders important and relevant to their curricula. However, there are limited time and practical opportunities for learning about eating disorders within current curricula. There is a need for guidance and resource development to help support educators to enhance eating disorder content in relevant curricula.