Mechanisms of G protein activation via the D-2 dopamine receptor: evidence for persistent receptor/G protein interaction after agonist stimulation

Background and purpose: The aim of this report is to study mechanisms of G protein activation by agonists. Experimental approach: The association and dissociation of guanosine 5'-O-(3-[S-35] thio) triphosphate ([S-35] GTP gamma S) binding at G proteins in membranes of CHO cells stably transfected with the human dopamine D-2short receptor was studied in the presence of a range of agonists. Key results: Binding of [S-35] GTPgS was dissociable in the absence of agonist and dissociation was accelerated both in rate and extent by dopamine, an effect which was blocked by the dopamine D-2 receptor antagonist raclopride and by suramin, which inhibits receptor/G protein interaction. A range of agonists of varying efficacy increased the rate of dissociation of [S-35] GTPgS binding, with the more efficacious agonists resulting in faster dissociation. Agonists were able to dissociate about 70% of the pre-bound [S-35] GTPgS, leaving a component which may not be accessible to the agonist-bound receptor. The dissociable component of the [S-35] GTPgS binding was reduced with longer association times and increased [S-35] GTPgS concentrations. Conclusions and implications: These data are consistent with [S-35] GTPgS binding being initially to receptor-linked G proteins and then to G proteins which have separated from the agonist bound receptor. Under the conditions used typically for [S-35] GTPgS binding assays, therefore, much of the agonist-receptor complex remains in proximity to G proteins after they have been activated by agonist.

Agonist-selective, receptor-specific interaction of human P-2Y receptors with beta-arrestin-1 and-2

Interaction of G-protein-coupled receptors with beta-arrestins is an important step in receptor desensitization and in triggering "alternative" signals. By means of confocal microscopy and fluorescence resonance energy transfer, we have investigated the internalization of the human P2Y receptors 1, 2, 4, 6, 11, and 12 and their interaction with beta-arrestin-1 and -2. Co-transfection of each individual P2Y receptor with beta-arrestin-1-GFP or beta-arrestin-2-YFP into HEK-293 cells and stimulation with the corresponding agonists resulted in a receptor-specific interaction pattern. The P2Y(1) receptor stimulated with ADP strongly translocated beta-arrestin-2-YFP, whereas only a slight translocation was observed for beta-arrestin-1-GFP. The P2Y(4) receptor exhibited equally strong translocation for beta-arrestin-1-GFP and beta-arrestin-2YFP when stimulated with UTP. The P2Y(6), P2Y(11), and P2Y(12) receptor internalized only when GRK2 was additionally cotransfected, but beta-arrestin translocation was only visible for the P2Y(6) and P2Y(11) receptor. The P2Y(2) receptor showed a beta-arrestin translocation pattern that was dependent on the agonist used for stimulation. UTP translocated beta-arrestin-1-GFP and beta-arrestin-2-YFP equally well, whereas ATP translocated beta-arrestin-1-GFP to a much lower extent than beta-arrestin2- YFP. The same agonist-dependent pattern was seen in fluorescence resonance energy transfer experiments between the fluorescently labeled P2Y(2) receptor and beta-arrestins. Thus, the P2Y(2) receptor would be classified as a class A receptor when stimulated with ATP or as a class B receptor when stimulated with UTP. The ligand-specific recruitment of beta-arrestins by ATP and UTP stimulation of P2Y(2) receptors was further found to result in differential stimulation of ERK phosphorylation. This suggests that the two different agonists induce distinct active states of this receptor that show differential interactions with beta-arrestins.

Drug interaction and location in liposomes: correlation with polar surface areas

An important step in liposome characterization is to determine the location of a drug within the liposome. This work thus investigated the interaction of dipalmitoylphosphatidylcholine liposomes with drugs of varied water solubility, polar surface area (PSA) and partition coefficient using high sensitivity differential scanning calorimetry. Lipophilic estradiol (ES) interacted strongest with the acyl chains of the lipid membrane, followed by the somewhat polar 5-fluorouracil (5-FU). Strongly hydrophilic mannitol (MAN) showed no evidence of interaction but water soluble polymers inulin (IN) and an antisense oligonucleotide (OLG), which have very high PSAs, interacted with the lipid head groups. Accordingly, the drugs could be classified as: hydrophilic ones situated in the aqueous core and which may interact with the head groups; those located at the water-bilayer interface with some degree of penetration into the lipid bilayer; those lipophilic drugs constrained within the bilayer. (c) 2004 Elsevier B.V. All rights reserved.

The effect of the functional attributes of objects within the caged environment on interaction time in laboratory rats

Determining rat preferences for, and behaviour towards, environmental enrichment objects allows us to provide evidence-based information about how the caged environment may be enriched. In recent years there have been many studies investigating the preferences of laboratory rodents for a wide variety of environmental enrichment objects and materials. While these have provided important information regarding the animals' perception of the items, very few studies have attempted to systematically investigate the precise attributes that constitute a preferred object and the behaviour that these objects afford. We have designed a research program to systematically study rats' motivation to interact with enrichment objects. Here we present the results from two experiments which examined the time rats spent with objects that only differed in size. This showed that rats spent longer with large objects rather than small ones, even though objects were presented individually. We also investigated the rats' behaviour with the objects in an open field and found that rats spent longer climbing on top of the large object. This behaviour continued when the large objects were laid on their sides instead of placed upright in the arena, suggesting that the rats were not simply climbing on the objects to investigate the top of the arena and thus an escape route, but instead were genuinely motivated to climb. This suggests that rat welfare could be enhanced by the addition to their cages of objects that permit climbing. (C) 2009 Elsevier B.V. All rights reserved.

Social interaction deficits and conversational inadequacy in Williams syndrome

Research on social communication skills in individuals with Williams syndrome has been inconclusive, with some arguing that these skills are a relative strength and others that they are a weakness. The aim of the present study was to investigate social interaction abilities in a group of children with WS, and to compare them to a group of children with specific language impairment and a group of typically developing children. Semi-structured conversations were conducted and 100-150 utterances were selected for analysis in terms of exchange structure, turn taking, information transfer and conversational inadequacy. The statistical analyses showed that the children with WS had difficulties with exchange structure and responding appropriately to the interlocutor's requests for information and clarification. They also had significant difficulties with interpreting meaning and providing enough information for the conversational partner. Despite similar language abilities with a group of children with specific language impairment, the children with WS had different social interaction skills, which suggests that they follow an atypical trajectory of development and their neurolinguistic profile does not directly support innate modularity. (c) 2005 Elsevier Ltd. All rights reserved.

The effect of postnatal depression on mother-infant interaction, infant response to the still-face perturbation, and performance on an instrumental learning task

A representative community sample of primiparous depressed women and a nondepressed control group were assessed while in interaction with their infants at 2 months postpartum. At 3 months, infants were assessed on the Still-face perturbation of face to face interaction, and a subsample completed an Instrumental Learning paradigm. Compared to nondepressed women, depressed mothers' interactions were both less contingent and less affectively attuned to infant behavior. Postnatal depression did not adversely affect the infant's performance in either the Still-face perturbation or the Instrumental Learning assessment. Maternal responsiveness in interactions at 2 months predicted the infant's performance in the Instrumental Learning assessment but not in the Still-face perturbation. The implications of these findings for theories of infant cognitive and emotional development are discussed.

Developing social interaction and understanding in individuals with autism spectrum disorder: a groupwork intervention

Background: Difficulties with social interaction and understanding lie at the heart of the communication disorder that characterises the autism spectrum. This study sought to improve social communication for individuals with autism spectrum disorder (ASD) by means of a groupwork intervention focusing on social and emotional perspective-taking, conversation skills, and friendship skills. It also aimed to address some of the limitations of previous interventions, including a lack of generalisation to other settings, so as to maximise inclusion in the community. Method A group of 46 high functioning children and adolescents with ASD (38 boys, 8 girls, age range 6-16 years) were allocated to one of 6 intervention groups. Each group met over a period of 12-16 weeks for a minimum of one 1 1/2-hour weekly session aimed at promoting key areas of social interaction and understanding, supported by home-based practice. Results: Significant gains were achieved in comparison with a normative population, and individual parent ratings showed marked and sustained changes in the key areas targeted in the group sessions. Conclusion: Social communication in children and adolescents with ASD can be enhanced through the use of a groupwork intervention addressing social interaction and understanding.

Sibling interaction of children with autism: development over 12 months

While deficits in social interaction are central to autism, the sibling relationship has been found to provide a key medium for the development of such skills. Naturalistic observations of sibling pairs including children with autism and controls with Down syndrome were made across two time periods, twelve months apart. Consistent with the evidence on typically developing children, the amount and rate of initiations of both prosocial and agonistic interaction increased, but further analysis suggested that these interactions were stage-managed by the typically developing children. Results show social interaction and imitation in children with autism and the special role that sibling interactions can play. Longitudinal research on the acquisition of social skills in children with developmental disabilities is needed.

Living with ASD: how do children and their parents assess their difficulties with social interaction and understanding?

Social interaction and understanding in autistic spectrum disorder (ASD) are key areas of concern to practitioners and researchers alike. However, there is a relative lack of information about the skills and competencies of children and young people with ASD who access ordinary community facilities including mainstream education. In particular, contributions by parents and their children have been under-utilized. Using two structured questionnaires, 19 children with ASD reported difficulties with social skills including social engagement and temper management and also reported difficulties with social competence, affecting both friendships and peer relationships. Parents rated the children's social skill and competence as significantly worse than did the children themselves, but there was considerable agreement about the areas that were problematic. Using an informal measure to highlight their children's difficulties, parents raised issues relating to conversation skills, social emotional reciprocity and peer relationships. The implications for assessment and intervention are discussed.

The role played by the interaction between genetic factors and attachment in the stress response in infancy

Background: The importance of understanding which environmental and biological factors are involved in determining individual differences in physiological response to stress is widely recognized, given the impact that stress has on physical and mental health. Methods: The child-mother attachment relationship and some genetic polymorphisms (5-HTTLPR, COMT and GABRA6) were tested as predictors of salivary cortisol and alpha amylase concentrations, two biomarkers of hypothalamic-pituitary-adrenocortical (HPA) axis and sympathetic adrenomedullary (SAM) system activity, during the Strange Situation (SS) procedure in a sample of more than 100 healthy infants, aged 12 to 18 months. Results: Individual differences in alpha amylase response to separation were predicted by security of attachment in interaction with 5-HTTLPR and GABRA6 genetic polymorphisms, whereas alpha amylase basal levels were predicted by COMT x attachment interaction. No significant effect of attachment, genetics and their interaction on cortisol activity emerged. Conclusions: These results help to disentangle the role played by both genetic and environmental factors in determining individual differences in stress response in infancy. The results also shed light on the suggestion that HPA and SAM systems are likely to have different characteristic responses to stress.

Object life cycles & interaction environments for cross media production & distribution

This paper addresses the requirements for a Work/flow Management System that is intended to automate the production and distribution chain for cross-media content which is by nature multi-partner and multi-site. It advocates the requirements for an ontology-based object lifecycle tracking within work/flow integration by identifying various types of interfaces, object life cycles and the work-flow interaction environments within the AXMEDIS Framework.

High bandwidth, large workspace haptic interaction: flying phantoms

It is well understood that for haptic interaction: free motion performance and closed-loop constrained motion performance have conflicting requirements. The difficulties for both conditions are compounded when increased workspace is required as most solutions result in a reduction of achievable impedance and bandwidth. A method of chaining devices together to increase workspace without adverse effect on performance is described and analysed. The method is then applied to a prototype, colloquially known as 'The Flying Phantom', and shown to provide high-bandwidth, low impedance interaction over the full range of horizontal movement across the front of a human user.

Robot-human interaction - practical experiments with a cyborg

This paper presents results to indicate the potential applications of a direct connection between the human nervous system and a computer network. Actual experimental results obtained from a human subject study are given, with emphasis placed on the direct interaction between the human nervous system and possible extra-sensory input. An brief overview of the general state of neural implants is given, as well as a range of application areas considered. An overall view is also taken as to what may be possible with implant technology as a general purpose human-computer interface for the future.