Isotope dilution liquid chromatography - tandem mass spectrometry for simultaneous identification and quantification of beta-casomorphin 5 and beta-casomorphin 7 in yoghurt

A highly selective and sensitive liquid chromatography–tandem mass spectrometry method was developed and validated for the simultaneous identification and quantification of beta-casomorphin 5 (BCM5) and beta-casomorphin 7 (BCM7) in yoghurt. The method used deuterium labelled BCM5-d10 and BCM7-d10 as surrogate standards for confident identification and accurate and quantification of these analytes in yoghurt. Linear responses for BCM5 and BCM7 (R2 = 0.9985 and 0.9986, respectively) was observed in the range 0.01–10 ng/μL. The method limits of detection (MLDs) in yoghurt extracts were found to be 0.5 and 0.25 ng/g for BCM5 and BCM7, respectively. Analyses of spiked samples were used to provide confirmation of accuracy and precision of the analytical method. Recoveries relative to the surrogate standards of these spikes were in the range of 95–106% for BCM5 and 103–109% for BCM7. Precision from analysis of spiked samples was expressed as relative standard deviation (%RSD) and values were in the range 1–16% for BCM5 and 1–6% for BCM7. Inter-day reproducibility was between 2.0–6.4% for BCM5 and between 3.2–6.1% for BCM7. The validated isotope dilution LC–MS/MS method was used to measure BCM5 and BCM7 in ten commercial and laboratory prepared samples of yoghurt and milk. Neither BCM5 nor BCM7 was detected in commercial yoghurts. However, they were observed in milk and laboratory prepared yoghurts and interestingly their levels decreased during processing. BCM5 decreased from 1.3 ng/g in milk to 1.1 ng/g in yoghurt made from that milk at 0 day storage and <MLQ at 1 and 7 days storage. BCM7 decreased from 1.9 ng/g in milk to <MLQ in yoghurts immediately after processing. These preliminary results indicate that fermentation and storage reduced BCM5 and BCM7 concentration in yoghurt

“Farm School : Recalling Meaning & Memory of the Lynn Bachman School

This project tells the story of a generation, a region, and a unique educational moment in Appalachian history through a series of hand printed, hand bound, illustrated artists books. Capturing the lived experience of elders in the communities of Polk County, Tennessee and Cherokee County, North Carolina, the research expands our understanding of Appalachian education and social work before public education was introduced to the area. The project appropriately merged Dockery’s interdisciplinary work in visual arts, art education and Appalachian Studies and expanded the scholarship on rural education. Utilizing an arts based research methodology, the research introduces the Lynn Bachman Memorial School and Home to the growing body of literature related to Appalachian Settlement Schools. Three elders were interviewed, an archives of historical and artistic images of the Bachman school was generated, and a limited edition of artists’ books that utilize the heirloom craft of letterpress, photo transfer, and German book binding technique was created

Identification of key structural elements for neuronal calcium sensor-1 function in the regulation of the temperature-dependency of locomotion in C. elegans

BACKGROUND: Intracellular Ca(2+) regulates many aspects of neuronal function through Ca(2+) binding to EF hand-containing Ca(2+) sensors that in turn bind target proteins to regulate their function. Amongst the sensors are the neuronal calcium sensor (NCS) family of proteins that are involved in multiple neuronal signalling pathways. Each NCS protein has specific and overlapping targets and physiological functions and specificity is likely to be determined by structural features within the proteins. Common to the NCS proteins is the exposure of a hydrophobic groove, allowing target binding in the Ca(2+)-loaded form. Structural analysis of NCS protein complexes with target peptides has indicated common and distinct aspects of target protein interaction. Two key differences between NCS proteins are the size of the hydrophobic groove that is exposed for interaction and the role of their non-conserved C-terminal tails. RESULTS: We characterised the role of NCS-1 in a temperature-dependent locomotion assay in C. elegans and identified a distinct phenotype in the ncs-1 null in which the worms do not show reduced locomotion at actually elevated temperature. Using rescue of this phenotype we showed that NCS-1 functions in AIY neurons. Structure/function analysis introducing single or double mutations within the hydrophobic groove based on information from characterised target complexes established that both N- and C-terminal pockets of the groove are functionally important and that deletion of the C-terminal tail of NCS-1 did not impair its ability to rescue. CONCLUSIONS: The current work has allowed physiological assessment of suggestions from structural studies on the key structural features that underlie the interaction of NCS-1 with its target proteins. The results are consistent with the notion that full length of the hydrophobic groove is required for the regulatory interactions underlying NCS-1 function whereas the C-terminal tail of NCS-1 is not essential. This has allowed discrimination between two potential modes of interaction of NCS-1 with its targets

Identification of key structural elements for neuronal calcium sensor-1 function in the regulation of the temperature-dependency of locomotion in C. elegans

BACKGROUND: Intracellular Ca2+ regulates many aspects of neuronal function through Ca2+ binding to EF hand-containing Ca2+ sensors that in turn bind target proteins to regulate their function. Amongst the sensors are the neuronal calcium sensor (NCS) family of proteins that are involved in multiple neuronal signalling pathways. Each NCS protein has specific and overlapping targets and physiological functions and specificity is likely to be determined by structural features within the proteins. Common to the NCS proteins is the exposure of a hydrophobic groove, allowing target binding in the Ca2+-loaded form. Structural analysis of NCS protein complexes with target peptides has indicated common and distinct aspects of target protein interaction. Two key differences between NCS proteins are the size of the hydrophobic groove that is exposed for interaction and the role of their non-conserved C-terminal tails. RESULTS: We characterised the role of NCS-1 in a temperature-dependent locomotion assay in C. elegans and identified a distinct phenotype in the ncs-1 null in which the worms do not show reduced locomotion at actually elevated temperature. Using rescue of this phenotype we showed that NCS-1 functions in AIY neurons. Structure/function analysis introducing single or double mutations within the hydrophobic groove based on information from characterised target complexes established that both N- and C-terminal pockets of the groove are functionally important and that deletion of the C-terminal tail of NCS-1 did not impair its ability to rescue. CONCLUSIONS: The current work has allowed physiological assessment of suggestions from structural studies on the key structural features that underlie the interaction of NCS-1 with its target proteins. The results are consistent with the notion that full length of the hydrophobic groove is required for the regulatory interactions underlying NCS-1 function whereas the C-terminal tail of NCS-1 is not essential. This has allowed discrimination between two potential modes of interaction of NCS-1 with its targets

The experiences of people with Alzheimer’s dementia and their caregivers in acquiring and using a mobility aid_ a qualitative study

Purpose: Cognitive deficits and gait and balance problems are progressive in people with Alzheimer’s dementia. Yet, mobility aids are associated with an increased falls risk in people with dementia. Our objectives were to identify the perceptions of people living with mild-to-moderate Alzheimer’s dementia, and their caregivers, on the use of mobility aids. Methods: A qualitative study using semi-structured, face-to-face interviews was conducted. Community-dwelling older adults with dementia attending a day hospital program were recruited. Thematic analysis was conducted and the text was coded into broad themes aligned with the research questions. The coded text was examined for patterns and similarities, and grouped to form inductive themes. Results: Twenty-four people (12 living with dementia and their 12 caregivers) participated. Five themes were identified: (1) acknowledgement of need; (2) protecting a sense of self; (3) caregiver oversight and relief of burden; (4) healthcare professional involvement; (5) environment and design of aids. Conclusions: The findings suggest that people with Alzheimer’s dementia and their caregivers regard mobility aid use as increasing independence. There is a role for healthcare professionals to be involved in the prescription, provision and training for use of mobility aids among people living with dementia to ensure uptake and safety.IMPLICATIONS FOR REHABILITATION Mobility aid use is regarded as increasing independence by people with Alzheimer’s disease and their caregivers. Falls risk associated with mobility aid use was not well known and caregivers perceived mobility aids as a means to reduce falls. Most people do not see a healthcare professional when they acquire a mobility aid and there is a role for healthcare professionals to be involved in the prescription, provision and training of people living with dementia in the use of mobility aids in order to increase the uptake of aids and their safe use

using provenance data and DNA

Re-examination of the historical range of the greater prairie chicke

Chimpanzee Rights: The Philosophers' Brief

In December 2013, the Nonhuman Rights Project (NhRP) filed a petition for a common law writ of habeas corpus in the New York State Supreme Court on behalf of Tommy, a chimpanzee living alone in a cage in a shed in rural New York (Barlow, 2017). Under animal welfare laws, Tommy’s owners, the Laverys, were doing nothing illegal by keeping him in those conditions. Nonetheless, the NhRP argued that given the cognitive, social, and emotional capacities of chimpanzees, Tommy’s confinement constituted a profound wrong that demanded remedy by the courts. Soon thereafter, the NhRP filed habeas corpus petitions on behalf of Kiko, another chimpanzee housed alone in Niagara Falls, and Hercules and Leo, two chimpanzees held in research facilities at Stony Brook University. Thus began the legal struggle to move these chimpanzees from captivity to a sanctuary, an effort that has led the NhRP to argue in multiple courts before multiple judges. The central point of contention has been whether Tommy, Kiko, Hercules, and Leo have legal rights. To date, no judge has been willing to issue a writ of habeas corpus on their behalf. Such a ruling would mean that these chimpanzees have rights that confinement might violate. Instead, the judges have argued that chimpanzees cannot be bearers of legal rights because they are not, and cannot be persons. In this book we argue that chimpanzees are persons because they are autonomous

It\u27s not too Late for the Harpy Eagle (Harpia harpyja): High Levels Of Genetic Diversity and Differentiation Can Fuel Conservation Programs

Background: The harpy eagle (Harpia harpyja) is the largest Neotropical bird of prey and is threatened by human persecution and habitat loss and fragmentation. Current conservation strategies include local education, captive rearing and reintroduction, and protection or creation of trans-national habitat blocks and corridors. Baseline genetic data prior to reintroduction of captive-bred stock is essential for guiding such efforts but has not been gathered previously. Methodology/Findings: We assessed levels of genetic diversity, population structure and demographic history for harpy eagles using samples collected throughout a large portion of their geographic distribution in Central America (n = 32) and South America (n = 31). Based on 417 bp of mitochondrial control region sequence data, relatively high levels of haplotype and nucleotide diversity were estimated for both Central and South America, although haplotype diversity was significantly higher for South America. Historical restriction of gene flow across the Andes (i.e. between our Central and South American subgroups) is supported by coalescent analyses, the haplotype network and significant FST values, however reciprocally monophyletic lineages do not correspond to geographical locations in maximum likelihood analyses. A sudden population expansion for South America is indicated by a mismatch distribution analysis, and further supported by significant (p,0.05) negative values of Fu and Li’s DF and F, and Fu’s FS. This expansion, estimated at approximately 60 000 years BP (99 000–36 000 years BP 95% CI), encompasses a transition from a warm and dry time period prior to 50 000 years BP to an interval of maximum precipitation (50 000–36 000 years BP). Notably, this time period precedes the climatic and habitat changes associated with the last glacial maximum. In contrast, a multimodal distribution of haplotypes was observed for Central America suggesting either population equilibrium or a recent decline. Significance: High levels of mitochondrial genetic diversity in combination with genetic differentiation among subgroups within regions and between regions highlight the importance of local population conservation in order to preserve maximal levels of genetic diversity in this species. Evidence of historically restricted female-mediated gene flow is an important consideration for captive-breeding programs