Characterisation of palmitoylation in alpha₂_A adrenoceptor and 5-HT₁_A serotonin receptor-G₀₁α G protein fusion proteins

Palmitoylation variant GPCR-G protein fusion proteins were created between the porcine u2A-adrenoceptor or the human 5-HT1A-serotonin receptor and the pertussis toxin resistant, Cys35lIle, form of the rat Go1u protein. These palmitoylation-variant fusions were transiently expressed in HEK293T cells prior to analysis of the regulation of palmitoylation and the functional consequences of palmitoylation for both the GPCR and G protein parts of the fusions. When the regulation of palmitoylation was studied for u2A-adrenoceptor-GoluCys35IIle fusion proteins, dynamic palmitoylation and depalmitoylation of both the Cys442residue of the u2A-adrenoceptor and the Cys ' residue of the GoluCys351Ile protein were found to occur. However, only the GOluCys351Ileprotein part of the fusion was found to undergo adrenaline-stimulated regulation of palmitoylation and the effect of adrenaline required G protein activation. Adrenaline regulation proceeded in a concentration-dependent manner correlating with agonist occupancy of the u2A-adrenoceptor. Such agonist effects were found to be, at least in part, due to agonist-stimulation of GOluCys351Ile protein depalmitoylation. The requirements for palmitoylation of the u2A-adrenoceptor and GoluCys351Ile protein elements of the u2A-adrenoceptor-GoluCys35IUe fusion proteins were subsequently assessed for various functional properties. Palmitoylation of neither the U2Aadrenoceptor nor the GoluCys351Ile protein parts of the fusion determined fusion protein expression levels, affinity for the agonist adrenaline, affinity for the antagonist RS- 79948-197, ability to bind or to hydrolyse GTP or their ability to influence the efficiency of RGS 16 protein to accelerate the GTPase reaction. In regulation of palmitoylation studies for 5-HTIA-receptor-GoluCys35IIle fusion proteins, dynamic palmitoylation of the Cys' residue of the GoluCys351Ue protein and the Cys417 residue of the 5-HTIA-receptor was observed as well as a lack of incorporation of palmitate into Cys420 of the 5-HT1A-receptor. Dynamic depalmitoylation was only observed for the Cys' residue of the GoluCys351Ile protein, not for the 5-HT1A-receptor. In the latter case, palmitate once incorporated appeared to remain stably attached. Both the 5-HT1A-receptor and the GoluCys351Ile protein parts of the fusion were found to undergo 8-0H-DPAT-stimulated regulation ofpalmitoylation. 8-0H-DPAT was able to regulate palmitoylation levels of both proteins in a concentration-dependent manner. For the regulation of GoluCys351Ile protein palmitoylation such agonist effects were found likely to be, at least in part, due to an agonist-stimulated rate of depalmitoylation. For the regulation of 5-HT1A-receptor palmitoylation such agonist-stimulated increases in observed palmitoylation levels were only attributable to the addition of palmitate, given that no depalmitoylation of the 5- HT1A-receptor could be detected. The requirements for palmitoylation of the 5-HT1A-receptor and GoluCys351Ile protein elements of the 5-HT1A-receptor-GoluCys351Ile fusion proteins were also assessed for a selection of functional properties. Similar to the results obtained with Go1uCys351Ile protein constrained to the uZA-adrenoceptor, the palmitoylation of the GoluCys351Ile protein did not determine fusion protein expression levels, their affinity for the antagonist WAYI00635, or their ability to bind GTP. Palmitoylation of 5-HT1Areceptor did not alter fusion protein expression levels or their affinity for the antagonist WAYI00635. However, in contrast, it did cause enhanced levels of GTP binding to the 5-HT1A-receptor-GoluCys351Ile fusion proteins. The results of this investigation suggest that there are different requirements for regulation of GPCR and G protein palmitoylation dependent on the GPCR-G protein fusion in question. These requirements may be responsible for the specific functional properties displayed by such fusions. The current study also demonstrates that GPCR-G protein fusion proteins can be successfully used as tools to study both the regulation of palmitoylation and the functional consequences of this modification

Domestic Violence Intervention Program Facilitators\u27 Motivation for Working With Repeat Offenders

Domestic violence (DV) rehabilitative program facilitators administer the same treatment programs to males who reoffend. When DV facilitators administer the same unsuccessful treatment programs to repeat offenders, facilitators may lose intrinsic and extrinsic motivation to perform their job. For this study a hermeneutic phenomenological methodology approach was used to explore the phenomenon of DV facilitators\u27 motivation. Self-determination theory was used to frame the influence of intrinsic and extrinsic motivation on DV facilitators who administer treatment programs to repeat offending males. A recruitment flyer was placed in the DV organization, data were collected from 7 participants through face-to-face or telephone interviews that were 18 years of age, proficient English speaking, actively facilitating DV treatment programs to repeat offending males, and employed with the DV organization. Data were transcribed and coded using open and axial coding, and analyzed for themes. Findings indicated that accountability, intrinsic motivation, and commitment influenced DV facilitators when administering programs to repeat offenders. Social change implications include awareness of perceived laissez-faire criminal justice policy towards DV male offenders and the lack of community support of challenges and opportunities for enhancing motivation for DV program facilitators

Evaluation of Indigenous justice programs Project D. Safe Aboriginal Youth Patrol programs in New South Wales and Northbridge policy and Juvenile Aid Group in Western Australia

In this evaluation, we examined two different approaches to delivery of Community and Night patrol services for young people: the Safe Aboriginal Youth Patrol programs (SAYP) of NSW, and the Northbridge Policy project (NPP) sometimes also called the Young people in Northbridge project, in Perth, Western Australia. The overarching focus of this evaluation was to determine whether the programs should be considered as examples of ‘good practice’ to be replicated elsewhere, and to find evidence of outcomes achieved by each program..

The Impact of Road Networks on Crime Rates in Saudi Arabia

This study examined the influence of the quality and extent of road networks on crime rates in both urban and rural areas of Jizan province, Saudi Arabia. We performed both Ordinary Least Squares regression (OLS) and Geographically Weighted Regression (GWR) where crime rate was the dependent variable and paved (sealed) roads, non-paved (unsealed/dirt) roads and population density were the explanatory variables. Population density was a control variable. The findings revealed that across all fourteen districts, those with better quality paved road networks recorded lower rates of crime than the districts with unpaved roads. Furthermore, the more extensive the road networks, the lower the crime rate, whether the roads were paved or not. The findings concur with those of studies conducted in other countries which reveal that rural areas are not always the safe, crime-free places they are often believed to be

Pawsitive Solutions: The symbiotic relationship between prisoners and dogs

With the prison population steadily increasing in Australia and over half of prisoners reoffending, it is evident that prison is ineffective for deterring and rehabilitating current and future offenders and reducing recidivism. As a result, there has been a gradual shift toward community corrections, placing an emphasis on interventions that address the criminogenic needs of prisoners. One such intervention is the implementation of prison dog programs (PDPs). PDPs involve a dog being paired with one or more specially selected inmates, who train, socialise and care for a dog for a specified period of time or until the animal is ready to be rehomed or move on to advanced training as an assistance or service dog. Although PDPs have been implemented in many correctional facilities in Australia, there is little evidence to support the existence of such programs. As such, this research sought to address this gap and add to the literature by conducting three studies. The first aimed to examine the nature and extent of PDPs operating in Australia through a national survey of eight corrections staff and 18 representatives from animal welfare, and training organisations involved in administering the program. The second study conducted semi-structured interviews with eight inmates, six corrections staff and one animal welfare representative involved in PDPs in Queensland, to identify the circumstances in which inmates are most likely to benefit from PDPs and how these programs can assist inmates in meeting their immediate and future needs. The third study aimed to explore the effect of PDPs on ten inmate participant's emotional intelligence; specifically, their ability to read emotions in others by comparing their ability, with current and previous dog owners, to provide judgements of emotion in photographs of dogs. The results of these studies support findings of other research and suggest that PDPs not only benefit inmates participating in PDPs, but also non-participant inmates, prison staff, prison culture, the dogs and society. The most reported benefits included positive changes to the prison environment, improved relationships with other inmates and staff and the opportunity to give back to society. The most commonly identified negative aspects were a lack of resources, personality clashes between inmates within the program and inmates' inability to socialise the dogs outside of the prison. Data from the studies as well as a review of the literature were used to develop a program logic model to improve the development, implementation, and evaluation of future PDPs

House time: Neolithic settlement development at Racot during the 5th millennium cal b.c. in the Polish lowlands

The settlement of Racot 18 in the western Polish lowlands is used as a case study in the investigation of continued development and expansion following initial Neolithic beginnings, and in the formal chronological modelling, in a Bayesian framework, of settlement development. The site belongs to the Late Lengyel culture of the later fifth millennium cal BC, and represents the intake of new land following earlier initial colonisation. The formally estimated chronology for the settlement suggests spans for individual house biographies from as little as a generation to over a century; distinctive substantial buildings, from late in the sequence, may have lasted longest. Racot 18 is compared to its formally modelled context of the later fifth millennium cal BC