Identification and proteomic profiling of exosomes in human cerebrospinal fluid

<p>Abstract</p> <p>Background</p> <p>Exosomes are released from multiple cell types, contain protein and RNA species, and have been exploited as a novel reservoir for disease biomarker discovery. They can transfer information between cells and may cause pathology, for example, a role for exosomes has been proposed in the pathophysiology of Alzheimer's disease. Although studied in several biofluids, exosomes have not been extensively studied in the cerebrospinal fluid (CSF) from humans. The objective of this study was to determine: 1) whether human CSF contains exosomes and 2) the variability in exosomal protein content across individuals.</p> <p>Methods</p> <p>CSF was collected from 5 study participants undergoing thoraco-abdominal aortic aneurysm repair (around 200 - 500 ml per participant) and low-density membrane vesicles were concentrated by ultracentrifugation. The presence of exosomes was determined by western blot for marker proteins, isopycnic centrifugation on a sucrose step gradient and transmission electron microscopy with immuno-labelling. Whole protein profiling was performed using Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR).</p> <p>Results</p> <p>Flotillin 1 and tumor susceptibility gene 101 (TSG101), two exosomal marker proteins, were identified in the ultracentrifugation pellet using western blot. These markers localized to a density consistent with exosomes following isopycnic centrifugation. Transmission electron microscopy visualized structures consistent with exosomes in size and appearance that labelled positive for flotillin 1. Therefore, the pellet that resulted from ultracentrifugation of human CSF contained exosomes. FT-ICR profiling of this pellet was performed and 84-161 ions were detected per study participant. Around one third of these ions were only present in a single study participant and one third were detected in all five. With regard to ion quantity, the median coefficient of variation was 81% for ions detected in two or more samples.</p> <p>Conclusions</p> <p>Exosomes were identified in human CSF and their proteome is a potential new reservoir for biomarker discovery in neurological disorders such as Alzheimer's disease. However, techniques used to concentrate exosomes from CSF need refinement to reduce variability. In this study we used relatively large starting volumes of human CSF, future studies will focus on exosome isolation from smaller 'real life' clinical samples; a key challenge in the development of exosomes as translational tools.</p

Development and initial validation of the bronchiectasis exacerbation and symptom tool (BEST)

BACKGROUND: Recurrent bronchiectasis exacerbations are related to deterioration of lung function, progression of the disease, impairment of quality of life, and to an increased mortality. Improved detection of exacerbations has been accomplished in chronic obstructive pulmonary disease through the use of patient completed diaries. These tools may enhance exacerbation reporting and identification. The aim of this study was to develop a novel symptom diary for bronchiectasis symptom burden and detection of exacerbations, named the BEST diary. METHODS: Prospective observational study of patients with bronchiectasis conducted at Ninewells Hospital, Dundee. We included patients with confirmed bronchiectasis by computed tomography, who were symptomatic and had at least 1 documented exacerbation of bronchiectasis in the previous 12\u2009months to participate. Symptoms were recorded daily in a diary incorporating cough, sputum volume, sputum colour, dyspnoea, fatigue and systemic disturbance scored from 0 to 26. RESULTS: Twenty-one patients were included in the study. We identified 29 reported (treated exacerbations) and 23 unreported (untreated) exacerbations over 6-month follow-up. The BEST diary score showed a good correlation with the established and validated questionnaires and measures of health status (COPD Assessment Test, r =\u20090.61, p =\u20090.0037, Leicester Cough Questionnaire, r =\u2009-\u20090.52,p =\u20090.0015, St Georges Respiratory Questionnaire, r =\u20090.61,p &lt;\u20090.0001 and 6\u2009min walk test, r =\u2009-\u20090.46,p =\u20090.037). The mean BEST score at baseline was 7.1 points (SD 2.2). The peak symptom score during exacerbation was a mean of 16.4 (3.1), and the change from baseline to exacerbation was a mean of 9.1 points (SD 2.5). Mean duration of exacerbations based on time for a return to baseline symptoms was 15.3\u2009days (SD 5.7). A minimum clinically important difference of 4 points is proposed. CONCLUSIONS: The BEST symptom diary has shown concurrent validity with current health questionnaires and is responsive at onset and recovery from exacerbation. The BEST diary may be useful to detect and characterise exacerbations in bronchiectasis clinical trials

Opinions of dentists and directors of nursing concerning dental care provision for Adelaide nursing homes

The document attached has been archived with permission from the Australian Dental Association (7th Jan 2007). An external link to the publisher’s copy is included.Background: The complex oral health problems of nursing home residents have been well documented. However, the influences on residents’ oral health status, including opinions and experiences of dental professionals and nursing home staff, have not yet been adequately investigated. Methods: The baseline questionnaire component of this longitudinal study was mailed to all registered dentists practising in Adelaide and Adelaide nursing home directors of nursing (DONs). Results: 413 dentists and 97 DONs indicated that Adelaide dentists’ interest and training in nursing home dentistry was low. Dental service provision for nursing home residents was very low and dentists preferred to provide treatment at their dental practices. Few dental hygienists were working in nursing homes and dental professionals provided little educational assistance for nursing home staff. Dentists and DONs held several common and many varying perceptions of the problems associated with dental care provision in nursing homes. Both identified a group of nursing home environmental constraints and a lack of portable dental equipment. DONs further identified a group of resident related problems, and dentists a group of dental practicerelated problems. Conclusions: These study results provide important information concerning problems with nursing home dentistry for dental service providers, educators, policy-makers, administrators and nursing home staff.JM Chalmers, C Hodge, JM Fuss, AJ Spencer, KD Carter and R Mathe

The prevalence and experience of oral diseases in Adelaide nursing home residents

The document attached has been archived with permission from the Australian Dental Association. An external link to the publisher’s copy is included.Background: The twenty-first century will see the evolution of a population of dentate older Australians with dental needs very different from those of older adults in past years. This study provided comprehensive information concerning oral disease prevalence in older South Australian nursing home residents. Methods: This paper presents cross-sectional baseline results. Results: Most of the 224 residents, from seven randomly selected nursing homes, were functionally dependent, medically compromised, cognitively impaired and behaviourally difficult older adults who presented many complex challenges to carers and to dental professionals. Two-thirds (66 per cent) were edentulous with many dental problems and treatment needs. Dentate residents had a mean of 11.9 teeth present, higher than previously reported. The prevalence and experience of coronal and root caries and plaque accumulation was very high in dentate residents; especially males, those admitted more than three years previously, those who ate fewer food types and those who were severely cognitively impaired. These residents had more retained roots, decayed teeth and missing teeth, and fewer filled teeth when compared with data for community-dwelling older adults. Conclusions: This study highlighted the poor oral health status of these nursing home residents and the great impact of dementia on their high levels of oral diseases.JM Chalmers, C Hodge, JM Fuss, AJ Spencer, KD Carte

Injuries, Ill-Health and Fatalities in White Water Rafting and White Water Paddling

White water (WW) activities such as paddling (canoeing and kayaking) and rafting are popular sports for recreational and professional participants. An increase in participation has been seen worldwide. However, these activities come with a risk of injury and even death if not conducted safely. A review was conducted to identify the types of injuries and ill-health that occur as a result of these activities. Injury and fatality rates were assessed to establish the risk attributed to these activities. Web of Science, PubMed, Ergonomics Abstracts and PsycINFO databases were searched and a total of 16 published articles were identified and reviewed. The shoulders and back were the most vulnerable sites for injury in WW paddling. Injuries to the face and lower limbs were most common in WW rafters. However, injury rates are low and estimates are discussed. Due to different methods used across the studies, the reported injury rates are not comparable. This review identified three illnesses incurred through WW activities. There may be more but these are not currently reported in the literature. A relative paucity of studies regarding injuries and fatalities in WW activities was identified. Directions for future research are suggested and discussed

Molecular characterisation of protist parasites in human-habituated mountain gorillas (Gorilla beringei beringei), humans and livestock, from Bwindi impenetrable National Park, Uganda

Over 60 % of human emerging infectious diseases are zoonotic, and there is growing evidence of the zooanthroponotic transmission of diseases from humans to livestock and wildlife species, with major implications for public health, economics, and conservation. Zooanthroponoses are of relevance to critically endangered species; amongst these is the mountain gorilla (Gorilla beringei beringei) of Uganda. Here, we assess the occurrence of Cryptosporidium, Cyclospora, Giardia, and Entamoeba infecting mountain gorillas in the Bwindi Impenetrable National Park (BINP), Uganda, using molecular methods. We also assess the occurrence of these parasites in humans and livestock species living in overlapping/adjacent geographical regions

To what extent can behaviour change techniques be identified within an adaptable implementation package for primary care? A prospective directed content analysis

Interpreting evaluations of complex interventions can be difficult without sufficient description of key intervention content. We aimed to develop an implementation package for primary care which could be delivered using typically available resources and could be adapted to target determinants of behaviour for each of four quality indicators: diabetes control, blood pressure control, anticoagulation for atrial fibrillation and risky prescribing. We describe the development and prospective verification of behaviour change techniques (BCTs) embedded within the adaptable implementation packages

The oral health assessment tool - Validity and reliability

The document attached has been archived with permission from the Australian Dental Association. An external link to the publisher’s copy is included.Background: The Oral Health Assessment Tool (OHAT) was a component of the Best Practice Oral Health Model for Australian Residential Care study. The OHAT provided institutional carers with a simple, eight category screening tool to assess residents’ oral health, including those with dementia. This analysis presents OHAT reliability and validity results. Methods: A convenience sample of 21 residential care facilities (RCFs) in urban and rural Victoria, NSW and South Australia used the OHAT at baseline, three-months and six-months to assess intra- and inter-carer reliability and concurrent validity. Results: Four hundred and fifty five residents completed all study phases. Intra-carer reliability for OHAT categories: percent agreement ranged from 74.4 per cent for oral cleanliness, to 93.9 per cent for dental pain; Kappa statistics were in moderate range (0.51-0.60) for lips, saliva, oral cleanliness, and for all other categories in range of 0.61-0.80 (substantial agreement) (p<0.05). Inter-carer reliability for OHAT categories: percent agreement ranged from 72.6 per cent for oral cleanliness to 92.6 per cent for dental pain; Kappa statistics were in moderate range (0.48-0.60) for lips, tongue, gums, saliva, oral cleanliness, and for all other categories in range of 0.61-0.80 (substantial agreement) (p<0.05). Intraclass correlation coefficients for OHAT total scores were 0.78 for intra-carer and 0.74 for inter-carer reliability. Validity analyses of the OHAT categories and examination findings showed complete agreement for the lips category, with the natural teeth, dentures, and tongue categories having high significant correlations and percent agreements. The gums category had significant moderate correlation and percent agreement. Non-significant and low correlations and percent agreements were evident for the saliva, oral cleanliness and dental pain categories. Conclusion: The Oral Health Assessment Tool was evaluated as being a reliable and valid screening assessment tool for use in residential care facilities, including those with cognitively impaired residents.JM Chalmers, PL King, AJ Spencer, FAC Wright and KD Carte

The ethics of inherent trust in care robots for the elderly

The way elderly care is delivered is changing. Attempts are being made to accommodate the increasing number of elderly, and the decline in the number of people available to care for them, with care robots. This change introduces ethical issues into robotics and healthcare. The two-part study (heuristic evaluation and survey) reported here examines a phenomenon which is a result of that change. The phenomenon rises out of a contradiction. All but 2 (who were undecided) of the 12 elderly survey respondents, out of the total of 102 respondents, wanted to be able to change how the presented care robot made decisions and 7 of those 12 elderly wanted to be able to examine its decision making process so as to ensure the care provided is personalized. However, at the same time, 34% of the elderly participants said they were willing to trust the care robot inherently, compared to only 16% of the participants who were under fifty. Additionally, 66% of the elderly respondents said they were very likely or likely to accept and use such a care robot in their everyday lives. The contradiction of inherent trust and simultaneous wariness about control gives rise to the phenomenon: elderly in need want control over their care to ensure it is personalized, but many may desperately take any help they can get. The possible causes, and ethical implications, of this phenomenon are the focus of this paper