Biochemical properties of mutant GBA on alpha-synuclein metabolism

Mutations in the GBA1 gene are numerically the greatest risk factor for Parkinson disease (PD). The GBA1 gene encodes for the lysosomal hydrolase enzyme, glucocerebrosidase (GCase), and mutations often lead to lysosomal impairment. The lysosome is important in the turnover of alpha-synuclein, the key hallmark for PD. The E326K mutation is one of the most common GBA1 mutations and is proposed to have a minimal effect on GCase activity. Common mutations, L444P and N370S, are well documented loss of function mutations. It was hypothesised that GBA1 mutations predispose patients to PD through influencing the relationship between GCase and alpha-synuclein, by altering the structural stability of GCase. The effect of the E326K mutation was characterised in human dermal fibroblasts, and demonstrated that it does not induce loss of GCase function, endoplasmic reticulum (ER) retention or ER stress, unlike L444P. Functional assays and proteolytic digestion analysis of recombinant GCase protein revealed minimal changes in stability across all mutants. This was corroborated in undifferentiated and differentiated SH-SY5Y dopaminergic neuron-like cell lines over-expressing GCase variants. Undifferentiated SH-SY5Y also demonstrated a drastic increase in lipid droplet number and a modest increase of insoluble alpha-synuclein aggregates in E326K mutants. Differentiated SH-SY5Y neurons and human midbrain dopamine neurons differentiated from induced pluripotent stem cells were treated with alpha-synuclein pre-formed fibrils (PFF) to accelerate alpha-synuclein pathology. No change in aggregation was demonstrated by Homogeneous Time Resolved Fluorescence® (HTRF®) analysis in E326K neurons compared to control. In conclusion, the work presented here demonstrates that the E326K mutation behaves differently to common loss of function mutations, however lipid dyshomeostasis and alpha-synuclein pathology is still evident

Predictors of obstetric anal sphincter injury during waterbirth: A secondary analysis of a prospective observational study

Introduction and Hypothesis: Obstetric Anal Sphincter Injury (OASI) during childbirth is associated with urino-genital pain and dysfunction. Waterbirth is a popular birth choice for women, but controversy remains around the risk of OASI during waterbirth. This study reports on the incidence of OASI, and factors associated with OASI, for a cohort of women who gave birth in water. Methods: This secondary analysis used prospectively collected data from 2,908 women who gave birth in water in the hospital setting. Incidence of OASI was calculated. Univariable and multivariable logistic regression analysis evaluated factors associated with OASI. Results: The incidence of OASI was 1.9% (95% CI 1.4, 2.4) for all women. In nulliparae it was higher (3.2%, 95% CI 2.3, 4.3), than for multiparae (0.9%, 95% CI 0.5, 1.4). In the multivariable analysis two variables were associated with OASI; multiparity was negatively associated with OASI (aOR 0.24, 95% CI 0.12, 0.50, p < 0.001), and birth weight was positively associated with OASI (aOR 1.001, 95% CI 1.000, 1.002, p = 0.02). A ‘hands-on’ technique was used during only 13% of births. Birth position supporting a flexible sacrum did not influence OASI risk. Conclusions: A low incidence of OASI was found for this cohort of women. The low proportion of midwives using a hands-on technique suggests that it may not be required in waterbirth.

Behavioural and physiological indicators of anxiety reflect shared and distinct profiles across individuals with neurogenetic syndromes

Anxiety is heightened in individuals with intellectual disability, particularly in those with specific neurogenetic syndromes. Assessment of anxiety for these individuals is hampered by a lack of appropriate measures that cater for communication impairment, differences in presentation, and overlapping features with co-occurring conditions. Here, we adopt a multi-method approach to identify fine-grained behavioural and physiological (via salivary cortisol) responses to anxiety presses in people with fragile X (FXS; n=27; Mage=20.11y; range 6.32-47.04y) and Cornelia de Lange syndromes (CdLS; n=27; Mage=18.42y; range 4.28–41.08y), two neurogenetic groups at high risk for anxiety, compared to neurotypical children (NT; n=21; Mage=5.97y, range 4.34–7.30y). Results indicate that physical avoidance of feared stimuli and proximity seeking to a familiar adult are prominent behavioural indicators of anxiety/stress in FXS and CdLS. Heightened pervasive physiological arousal was identified in these groups via salivary cortisol. An association between autistic characteristics and anxiety was evident in the FXS group but not in the CdLS group pointing to syndrome-specific nuances in the association between anxiety and autism. This study furthers understanding of the behavioural and physiological presentation of anxiety in individuals with intellectual disability and progresses theoretical developments regarding the development and maintenance of anxiety at the intersection of autism

Prognostic variables and scores identifying the end of life in COPD: a systematic review.

INTRODUCTION: COPD is a major cause of mortality, and the unpredictable trajectory of the disease can bring challenges to end-of-life care. We aimed to investigate known prognostic variables and scores that predict prognosis in COPD in a systematic literature review, specifically including variables that contribute to risk assessment of patients for death within 12 months. METHODS: We conducted a systematic review on prognostic variables, multivariate score or models for COPD. Ovid MEDLINE, EMBASE, the Cochrane database, Cochrane CENTRAL, DARE and CINAHL were searched up to May 1, 2016. RESULTS: A total of 5,276 abstracts were screened, leading to 516 full-text reviews, and 10 met the inclusion criteria. No multivariable indices were developed with the specific aim of predicting all-cause mortality in stable COPD within 12 months. Only nine indices were identified from four studies, which had been validated for this time period. Tools developed using expert knowledge were also identified, including the Gold Standards Framework Prognostic Indicator Guidance, the RADboud Indicators of Palliative Care Needs, the Supportive and Palliative Care Indicators Tool and the Necesidades Paliativas program tool. CONCLUSION: A number of variables contributing to the prediction of all-cause mortality in COPD were identified. However, there are very few studies that are designed to assess, or report, the prediction of mortality at or less than 12 months. The quality of evidence remains low, such that no single variable or multivariable score can currently be recommended

Developing doctors: what are the attitudes and perceptions of year 1 and 2 medical students towards a new integrated formative objective structured clinical examination?

Abstract Background: Objective Structured Clinical Examination (OSCE) is a core component of undergraduate medical student assessment. With increased emphasis on integrated programmes, more courses include OSCEs in the traditionally &apos;preclinical&apos; years. The acceptability and impact of such assessment methods at this stage of training is unknown

Skill set or mind set? Associations between health literacy, patient activation and health.

OBJECTIVE: There is ongoing debate on whether health literacy represents a skill-based construct for health self-management, or if it also more broadly captures personal 'activation' or motivation to manage health. This research examines 1) the association between patient activation and health literacy as they are most commonly measured and 2) the independent and combined associations of patient activation and health literacy skills with physical and mental health. METHODS: A secondary analysis of baseline cross-sectional data from the LitCog cohort of older adults was used. Participants (n = 697) were recruited from multiple US-based health centers. During structured face-to-face interviews, participants completed the Test of Functional Health Literacy in Adults (TOFHLA), the Patient Activation Measure (PAM), the SF-36 physical health summary subscale, and Patient Reported Outcomes Measurement Information Service (PROMIS) short form subscales for depression and anxiety. RESULTS: The relationship between health literacy and patient activation was weak, but significant (r = 0.11, p<0.01). In models adjusted for participant characteristics, lower health literacy was associated with worse physical health (β = 0.13, p<0.001) and depression (β = -0.16, p<0.001). Lower patient activation was associated with worse physical health (β = 0.19, p<0.001), depression (β = -0.27, p<0.001) and anxiety (β-0.24, p<0.001). CONCLUSIONS: The most common measures of health literacy and patient activation are weakly correlated with each other, but also independently correlated with health outcomes. This suggests health literacy represents a distinct skill-based construct, supporting the Institute of Medicine's definition. Deficits in either construct could be useful targets for behavioral intervention

Effects of Harmful Algal Blooms on Fish and Shellfish Species: A Case Study of New Zealand in a Changing Environment

Harmful algal blooms (HABs) have wide-ranging environmental impacts, including on aquatic species of social and commercial importance. In New Zealand (NZ), strategic growth of the aquaculture industry could be adversely affected by the occurrence of HABs. This review examines HAB species which are known to bloom both globally and in NZ and their effects on commercially important shellfish and fish species. Blooms of Karenia spp. have frequently been associated with mortalities of both fish and shellfish in NZ and the sub-lethal effects of other genera, notably Alexandrium spp., on shellfish (which includes paralysis, a lack of byssus production, and reduced growth) are also of concern. Climate change and anthropogenic impacts may alter HAB population structure and dynamics, as well as the physiological responses of fish and shellfish, potentially further compromising aquatic species. Those HAB species which have been detected in NZ and have the potential to bloom and harm marine life in the future are also discussed. The use of environmental DNA (eDNA) and relevant bioassays are practical tools which enable early detection of novel, problem HAB species and rapid toxin/HAB screening, and new data from HAB monitoring of aquaculture production sites using eDNA are presented. As aquaculture grows to supply a sizable proportion of the world’s protein, the effects of HABs in reducing productivity is of increasing significance. Research into the multiple stressor effects of climate change and HABs on cultured species and using local, recent, HAB strains is needed to accurately assess effects and inform stock management strategies

Scribble modulates the MAPK/Fra1 pathway to disrupt luminal and ductal integrity and suppress tumour formation in the mammary gland

Polarity coordinates cell movement, differentiation, proliferation and apoptosis to build and maintain complex epithelial tissues such as the mammary gland. Loss of polarity and the deregulation of these processes are critical events in malignant progression but precisely how and at which stage polarity loss impacts on mammary development and tumourigenesis is unclear. Scrib is a core polarity regulator and tumour suppressor gene however to date our understanding of Scrib function in the mammary gland has been limited to cell culture and transplantation studies of cell lines. Utilizing a conditional mouse model of Scrib loss we report for the first time that Scrib is essential for mammary duct morphogenesis, mammary progenitor cell fate and maintenance, and we demonstrate a critical and specific role for Scribble in the control of the early steps of breast cancer progression. In particular, Scrib-deficiency significantly induced Fra1 expression and basal progenitor clonogenicity, which resulted in fully penetrant ductal hyperplasia characterized by high cell turnover, MAPK hyperactivity, frank polarity loss with mixing of apical and basolateral membrane constituents and expansion of atypical luminal cells. We also show for the first time a role for Scribble in mammalian spindle orientation with the onset of mammary hyperplasia being associated with aberrant luminal cell spindle orientation and a failure to apoptose during the final stage of duct tubulogenesis. Restoring MAPK/Fra1 to baseline levels prevented Scrib-hyperplasia, whereas persistent Scrib deficiency induced alveolar hyperplasia and increased the incidence, onset and grade of mammary tumours. These findings, based on a definitive genetic mouse model provide fundamental insights into mammary duct maturation and homeostasis and reveal that Scrib loss activates a MAPK/Fra1 pathway that alters mammary progenitor activity to drive premalignancy and accelerate tumour progression