An improved protocol for the isolation of total genomic DNA from Labyrinthulomycetes

Many protocols have been used for extraction of DNA from Thraustochytrids. These generally involve the use of CTAB, phenol/chloroform and ethanol. They also feature mechanical grinding, sonication, N2 freezing or bead beating. However, the resulting chemical and physical damage to extracted DNA reduces its quality. The methods are also unsuitable for large numbers of samples. Commercially-available DNA extraction kits give better quality and yields but are expensive. Therefore, an optimized DNA extraction protocol was developed which is suitable for Thraustochytrids to both minimise expensive and time-consuming steps prior to DNA extraction and also to improve the yield. The most effective method is a combination of single bead in TissueLyser (Qiagen) and Proteinase K. Results were conclusive: both the quality and the yield of extracted DNA were higher than with any other method giving an average yield of 8.5 µg/100 mg biomass.

Manufacturing and use of human placenta-derived mesenchymal stromal cells for phase I clinical trials: Establishment and evaluation of a protocol

Background/Aim. Mesenchymal stromal cells (MSCs) have been utilised in many clinical trials as an experimental treatment in numerous clinical settings. Bone marrow remains the traditional source tissue for MSCs but is relatively hard to access in large volumes. Alternatively, MSCs may be derived from other tissues including the placenta and adipose tissue. In an initial study no obvious differences in parameters such as cell surface phenotype, chemokine receptor display, mesodermal differentiation capacity or immunosuppressive ability, were detected when we compared human marrow derived- MSCs to human placenta-derived MSCs. The aim of this study was to establish and evaluate a protocol and related processes for preparation placenta-derived MSCs for early phase clinical trials. Methods. A full-term placenta was taken after delivery of the baby as a source of MSCs. Isolation, seeding, incubation, cryopreservation of human placentaderived MSCs and used production release criteria were in accordance with the complex regulatory requirements applicable to Code of Good Manufacturing Practice manufacturing of ex vivo expanded cells. Results. We established and evaluated instructions for MSCs preparation protocol and gave an overview of the three clinical areas application. In the first trial, MSCs were co-transplanted iv to patient receiving an allogeneic cord blood transplant as therapy for treatmentrefractory acute myeloid leukemia. In the second trial, MSCs were administered iv in the treatment of idiopathic pulmonary fibrosis and without serious adverse effects. In the third trial, MSCs were injected directly into the site of tendon damage using ultrasound guidance in the treatment of chronic refractory tendinopathy. Conclusion. Clinical trials using both allogeneic and autologous cells demonstrated MSCs to be safe. A described protocol for human placenta-derived MSCs is appropriate for use in a clinical setting, relatively inexpensive and can be relatively easily adjusted to a different set of regulatory requirements, as applicable to early phase clinical trials.

Health-related quality of life of individuals with transfemoral amputation fitted with the Transcutaneous Bone Anchoring Prosthesis following the OGAAP

Background The benefits and safety transcutaneous bone anchored prosthesis relying on a screw fixation are well reported.[1-17] However, most of the studies on press-fit implants and joint replacement technology have focused on surgical techniques.[3, 18-23] One European centre using this technique has reported on health related quality of life (HRQOL) for a group of individuals with transfemoral amputation (TFA).[3] Data from other centres are needed to assess the effectiveness of the technique in different settings. Aim This study aimed at reporting HRQOL data at baseline and up to 2-year follow-up for a group of TFAs treated by Osseointegration Group of Australia who followed the Osseointegration Group of Australia Accelerated Protocol (OGAAP), in Sydney between 08/12/2011 and 09/04/2014. Method A total of 16 TFAs (7 females and 9 males, age 51 ± 12 y, height 1.73 ± 0.12 m, weight 83 ±18 kg) participated in this study. The cause of amputation was trauma or congenital limb deficiency for 11 (69%) and 5 (31%) participants, respectively. A total of 12 (75%) participants were prosthetic users while 4(25%) were wheelchair bound prior the surgery. The HRQOL were obtained from Questionnaire for Persons with Transfemoral Amputation (Q-TFA) using the four main scales (i.e., Prosthetic use, Mobility, Problem, Global) one year before and between 6.5 and 24 months after the Stage 1 of the surgeries for the baseline and follow-up, respectively. Results The lapse of time before and after Stage 1 was -6.19±3.54 and 10.83±3.58 months respectively. The raw score and percentage of improvement are presented in Figures 1 and 2, respectively. Discussion & Conclusion The average results demonstrated an improvement in each domain, particularly in the reduction of problems and an increase in global state. Furthermore, 56%, 75%, 94% and 69% of the participants reported an improvement in Prosthetic use, Mobility, Problem, Global scales, respectively. These results were comparable to previous studies relying of screwed fixation confirming that press-fit implantation is a viable alternative for bone-anchored prostheses.[1, 7, 8]

The thrill of victory: Women and sport shopping

This study explores a new shopper type, ‘Sport Shoppers’, who are unique in their actions, attitudes and behaviors. We present evidence that these shoppers exhibit behavior similar to that of competitive athletes in that they view bargain shopping as an achievement domain. Data were collected through open-ended surveys, in-depth interviews and closet inventories. Thematic analysis of transcripts was undertaken based on three achievement goal categories; task-oriented, ego-oriented and social approval-oriented. Achievement goal theory is employed to offer an explanation of the Sport Shoppers’ behaviors and motivations. We argue the Sport Shopper is an economically viable and important shopper for fashion retailers to target.

How to choose a healthy, eco-friendly childcare centre

The early years have been recognized as a “natural starting point” for education for sustainability. ECEfS is a newly emerging area and Australian educators, teacher educators, and researchers are leaders in this field. Indeed, there is a growing list of exemplary kindergartens, child care centres, preschools, and preps across the country engaged in EfS focusing on a range of topics that include, for example, water education, reducing energy use, recycling, growing gardens, and cleaning up parks and creeks. Some centres focus more on the social dimensions of sustainability – strengthening Indigenous communities, working with the elderly, supporting the unemployed, fostering gender equality in their communities, being more socially inclusive. There is mounting evidence that early lessons in sustainability help reshape adult sustainability actions in homes, schools and communities.

Solvent effect in molecular recognition: Determining binding constants in different solvents following an extraction based protocol

Semi-rigid molecular tweezers 1, 3 and 4 bind picric acid with more than tenfold increment in tetrachloromethane as compared to chloroform.

PocketMatch: A new algorithm to compare binding sites in protein structures

Recognizing similarities and deriving relationships among protein molecules is a fundamental requirement in present-day biology. Similarities can be present at various levels which can be detected through comparison of protein sequences or their structural folds. In some cases similarities obscure at these levels could be present merely in the substructures at their binding sites. Inferring functional similarities between protein molecules by comparing their binding sites is still largely exploratory and not as yet a routine protocol. One of the main reasons for this is the limitation in the choice of appropriate analytical tools that can compare binding sites with high sensitivity. To benefit from the enormous amount of structural data that is being rapidly accumulated, it is essential to have high throughput tools that enable large scale binding site comparison. Results: Here we present a new algorithm PocketMatch for comparison of binding sites in a frame invariant manner. Each binding site is represented by 90 lists of sorted distances capturing shape and chemical nature of the site. The sorted arrays are then aligned using an incremental alignment method and scored to obtain PMScores for pairs of sites. A comprehensive sensitivity analysis and an extensive validation of the algorithm have been carried out. A comparison with other site matching algorithms is also presented. Perturbation studies where the geometry of a given site was retained but the residue types were changed randomly, indicated that chance similarities were virtually non-existent. Our analysis also demonstrates that shape information alone is insufficient to discriminate between diverse binding sites, unless combined with chemical nature of amino acids. Conclusion: A new algorithm has been developed to compare binding sites in accurate, efficient and high-throughput manner. Though the representation used is conceptually simplistic, we demonstrate that along with the new alignment strategy used, it is sufficient to enable binding comparison with high sensitivity. Novel methodology has also been presented for validating the algorithm for accuracy and sensitivity with respect to geometry and chemical nature of the site. The method is also fast and takes about 1/250(th) second for one comparison on a single processor. A parallel version on BlueGene has also been implemented.

More than just food: Field visits to an emergency relief centre

Emergency relief centres provide financial, housing, food and other types of support to families and individuals who experience financial hardship. These centres are non-profit, often government supported organizations that rely on the help of their volunteers and social workers. This paper reports on our preliminary findings from field visits to one such centre called Communify, in the inner west of Brisbane, Australia. Communify runs an emergency food relief facility for people who find themselves in a crisis or temporarily unable to afford groceries. Over a period of five months, we did several field visits to the centre and carried out 21 short in-situ interviews, with a mix of Communify clients and volunteers. Our results shed light on people’s experiences of financial hardship and their interactions with the emergency relief centre. In particular, issues related to their perceived values and stigmas associated with their experiences are highlighted in our findings. We identify opportunities for design that can empower people struggling with financial hardship.

G(its)(2) VSR: An Information Theoretic Secure Verifiable Secret Redistribution Protocol for Long-term Archival Storage

Protocols for secure archival storage are becoming increasingly important as the use of digital storage for sensitive documents is gaining wider practice. Wong et al.[8] combined verifiable secret sharing with proactive secret sharing without reconstruction and proposed a verifiable secret redistribution protocol for long term storage. However their protocol requires that each of the receivers is honest during redistribution. We proposed[3] an extension to their protocol wherein we relaxed the requirement that all the recipients should be honest to the condition that only a simple majority amongst the recipients need to be honest during the re(distribution) processes. Further, both of these protocols make use of Feldman's approach for achieving integrity during the (redistribution processes. In this paper, we present a revised version of our earlier protocol, and its adaptation to incorporate Pedersen's approach instead of Feldman's thereby achieving information theoretic secrecy while retaining integrity guarantees.

Submission to the External Independent Review of the United Nations Response to Allegations of Sexual Exploitation and Abuse and Other Serious Crimes by Members of Foreign Military Forces Not Under United Nations Command in the Central African Republic

In 2015 the UN Secretary-General established an External Independent Review to review how the United Nations has responded to allegations of child sexual exploitation and child sexual abuse, and to make recommendations concerning how the United Nations should respond to allegations in the future. This submission to the Review Panel draws on literature regarding children's rights, the nature of child sexual abuse, international instruments and policy, the nature of institutional child sexual abuse, and the CAR case itself. It makes recommendations for reform of UN protocols and procedures to better prevent child sexual abuse, and to improve responses to future occurrences.

The effectiveness of peer-based interventions on health promoting behaviors in older people: A systematic review protocol of quantitative evidence

Peer-based interventions have the potential to enhance quality of life and functioning; however their role specifically within the older population has not been fully investigated. The objective of this review therefore is to locate, appraise and synthesise evidence on the effectiveness of peer-based interventions on changes in health behaviors, specifically for the older population. The specific question to be answered is: “what is the effectiveness of peer-based interventions on health promoting behaviors in older adults, when compared to non peer-based interventions?”

Family violence in culturally and linguistically diverse communities: An evaluation of a family relationship centre

Federal legislative changes in Australia have sought to improve how family relationship centres (FRCs) can be more responsive to culturally and linguistically diverse (CALD) communities when addressing family disputes and family violence. Research on the prevalence of family violence against women from CALD backgrounds is sparse. This article seeks to contribute to the understanding of this issue by describing findings of an evaluation of the FRC at Broadmeadows conducted by the authors. The findings focus on family violence and the appropriateness of the services offered to three CALD community groups, namely Iraqi, Lebanese and Turkish. The final section provides reflections on these findings and offers suggestions about how FRC services might better cater to the family violence needs of CALD communities by developing more extensive partnerships with community groups and by expanding the range of processes they offer beyond mediation.

Effect of an interactive therapeutic robotic animal on engagement, mood states, agitation and psychotropic drug use in people with dementia: A cluster-randomised controlled trial protocol

Introduction: Apathy, agitated behaviours, loneliness and depression are common consequences of dementia. This trial aims to evaluate the effect of a robotic animal on behavioural and psychological symptoms of dementia in people with dementia living in long-term aged care. Methods and analysis: A cluster-randomised controlled trial with three treatment groups: PARO (robotic animal), Plush-Toy (non-robotic PARO) or Usual Care (Control). The nursing home sites are Australian Government approved and accredited facilities of 60 or more beds. The sites are located in South-East Queensland, Australia. A sample of 380 adults with a diagnosis of dementia, aged 60 years or older living in one of the participating facilities will be recruited. The intervention consists of three individual 15 min non-facilitated sessions with PARO or Plush- Toy per week, for a period of 10 weeks. The primary outcomes of interest are improvement in agitation, mood states and engagement. Secondary outcomes include sleep duration, step count, change in psychotropic medication use, change in treatment costs, and staff and family perceptions of PARO or Plush-Toy. Video data will be analysed using Noldus XT Pocket Observer; descriptive statistics will be used for participants’ demographics and outcome measures; cluster and individual level analyses to test all hypotheses and Generalised Linear Models for cluster level and Generalised Estimation Equations and/or Multi-level Modeling for individual level data. Ethics and dissemination: The study participants or their proxy will provide written informed consent. The Griffith University Human Research Ethics Committee has approved the study (NRS/03/14/HREC). The results of the study will provide evidence of the efficacy of a robotic animal as a psychosocial treatment for the behavioural and psychological symptoms of dementia. Findings will be presented at local and international conference meetings and published in peer-reviewed journals.

Ionized Gas towards Galactic Centre-Constraints from Low-Frequency Recombination Lines

Observations of the H272α recombination line towards the galactic centre show features near VLSR= 0, -50 and + 36 kms-1 . We have combined the parameters of these features with the available -166 measurements to obtain the properties of the ionized gas present along the line of sight and also in the -3 kpc arm-. For the line-of-sight ionized gas we get an electron density around 7 cm-3 and a pathlength through it 10-60 pc. The emission measure and the electron temperature are in the range 500-2900 pc cm-6 and 2000-6000 - respectively. The ionized gas in the 3 kpc arm has an electron density of 30 cm-3 and extends over 9 pc along the line of sight if we assume an electron temperature of 104 K. Using the available upper limit to the intensity of the H351α recombination line, we show that the distributed ionized gas responsible for the dispersion of pulsar signals should have a temperature > 4500 - and a minimum filling factor of 20 per cent. We also show that recombination lines from the -warm ionized- gas proposed by McKee & Ostriker (1977) should be detectable in the frequency range 100-150 MHz towards the galactic centre with the sensitivity available at present.

Analysis of an inter-centre, web-based radiation oncology peer-review case conference

Purpose Peer-review programmes in radiation oncology are used to facilitate the process and evaluation of clinical decision-making. However, web-based peer-review methods are still uncommon. This study analysed an inter-centre, web-based peer-review case conference as a method of facilitating the decision-making process in radiation oncology. Methodology A benchmark form was designed based on the American Society for Radiation Oncology targets for radiation oncology peer review. This was used for evaluating the contents of the peer-review case presentations on 40 cases, selected from three participating radiation oncology centres. A scoring system was used for comparison of data, and a survey was conducted to analyse the experiences of radiation oncology professionals who attended the web-based peer-review meetings in order to identify priorities for improvement. Results The mean scores for the evaluations were 82·7, 84·5, 86·3 and 87·3% for cervical, prostate, breast and head and neck presentations, respectively. The survey showed that radiation oncology professionals were confident about the role of web-based peer-reviews in facilitating sharing of good practice, stimulating professionalism and promoting professional growth. The participants were satisfied with the quality of the audio and visual aspects of the web-based meeting. Conclusion The results of this study suggest that simple inter-centre web-based peer-review case conferences are a feasible technique for peer review in radiation oncology. Limitations such as data security and confidentiality can be overcome by the use of appropriate structure and technology. To drive the issues of quality and safety a step further, small radiotherapy departments may need to consider web-based peer-review case conference as part of their routine quality assurance practices.