COGKNOW development and evaluation of an ICT-device for people with mild dementia.

Dementia is a progressive, chronic disease affecting 5% of all persons above 65 and over 40% of people over 90. The aim of the COGKNOW project is to achieve a breakthrough with research that addresses the needs of those with dementia, particularly those with mild dementia living in the community. This entails cognitive reinforcement in four main areas: helping people to remember, helping to maintain social contact, helping with performing daily life and recreational activities and finally enhance feelings of safety. Based on a sound foundation of needs reported in dementia literature, workshops and individual interviews have been carried out with dementia sufferers and their carers in three European countries. A ranked analysis of information from workshops and interviews, and the state of the art of successful ICT solutions will be the basis for formulating the functionalities of the technical solution and for the development of a cognitive prosthetic device with associated services for people with mild dementia. The research and evaluation will be conducted from human factors, technology, and business perspectives in three phases of one year each. In this paper we discuss the design of the COGKNOW project, the first results of the user needs inquiry workshops and the ICT solutions the COGKNOW project will focus on in the first year.

Induction of circular episomes during rescue and replication of adeno-associated virus in experimental models of virus latency.

The synthesis of linear duplex replicative structures (monomers, head-to-head, and tail-to-tail dimers) is an important hallmark of the productive phase of the adeno-associated virus (AAV) life cycle. These structures are generated by a strand-displacement replication mechanism and believed to be a reservoir for single-stranded DNA genomes. During the course of studies with recombinant versions of AAV (rAAV), we discovered the assembly of circular duplex provirus derivatives in latently infected cell lines under conditions permissive for replication (i.e., helper virus dependent). These novel structures were cloned by bacterial trapping revealing a markedly homogeneous structure that included a single copy of the rAAV genome joined head-to-tail about the inverted terminal repeats (ITR). Restriction and sequence analysis of the point of circularization revealed a so-called "TRT" domain, consisting of a single ITR hairpin palindrome flanked by 5' and 3' D sequence elements. The circular conformation was additionally characterized by Southern blotting and confirmed by purification on an ethidium bromide-CsCl gradient where the buoyant density was consistent with circular supercoiled DNA. These findings suggest that AAV replication is accompanied by the assembly of circular duplex structures.

Interactions of forskolin and ATP with the cytosolic domains of mammalian adenylyl cyclase.

Fragments of the two cytoplasmic domains of mammalian adenylyl cyclases can be synthesized independently (and abundantly) as soluble proteins; Gsalpha- and forskolin-stimulated enzymatic activity is restored upon their mixture. We have utilized this system to characterize the interactions of adenylyl cyclase with forskolin and its substrate, ATP. In the presence of Gsalpha, adenylyl cyclase is activated in response to occupation of only one forskolin-binding site. A single binding site for forskolin was identified by equilibrium dialysis; its Kd (0.1 microM) corresponds to the EC50 for enzyme activation. The affinity of forskolin for adenylyl cyclase is greatly reduced in the absence of Gsalpha ( approximately 40 microM). Binding of forskolin to the individual cytoplasmic domains of the enzyme was not detected. A single binding site for the ATP analog, alpha,beta-methylene ATP (Ap(CH2)pp), was also detected by equilibrium dialysis. Such binding was not observed with the individual domains. Binding of Ap(CH2)pp was unaffected by P-site inhibitors of adenylyl cyclase. A modified P-loop sequence located near the carboxyl terminus of adenylyl cyclase has been implicated in ATP binding. Mutation of the conserved, non-glycine residues within this region caused no significant changes in the Km for ATP or the Ki for Ap(CH2)pp. It thus seems unlikely that this region is part of the active site. However, a mutation in the C1 domain (E518A) causes a 10-fold decrease in the binding affinity for Ap(CH2)pp. This residue and the active site of the enzyme may lie at the interface between the two cytosolic domains.

Repair of transversely incised abdominal wall fascia in a rabbit model.

OBJECTIVE: To compare several commonly used methods of closing transversely incised anterior abdominal wall in order to determine which technique results in the strongest incisional tensile strength. METHODS: Thirty-six rabbits were randomized to receive either interrupted or continuous closure with 0-Vicryl. Within these groups, each animal was randomized to one of three different bite and interval techniques: 1-cm bites/0.5-cm intervals, 1-cm bites/1-cm intervals, and 2-cm bites/1-cm intervals. Each rabbit received three to four transverse abdominal wall incisions of approximately 3-8 cm in length. The incisions were excised en bloc and stored at -70C at postoperative week 1, 2, or 4 in a random fashion. Representative 1-cm strips were harvested from each incision after thawing. The Instron tensiometer was used to determine the maximum intrinsic tensile strength required to disrupt each tissue strip at the incision. Statistical analysis was performed using analysis of variance, two-sample t test, Scheffé multiple comparison, and Kruskal-Wallis test. RESULTS: Two hundred thirty-seven strips were analyzed. The mean maximum tensile strength of all of the interrupted and continuous suture repairs was 48 and 38 lb, respectively (P < .001). The maximum tensile strength for interrupted closures was achieved at week 1 and was similar at week 4. The continuous closure was weakest at week 1 and increased to a maximum value during week 4. There was no difference in maximum tensile strength between the interrupted and continuous closure groups at week 4. There was no significant difference in the maximum tensile strength of the three repair techniques. The mean maximum tensile strength of all specimens was significantly less among those harvested during weeks 1 and 2 compared with week 4 (P = .001). CONCLUSION: In this randomized study, the interrupted closure had a greater maximum tensile strength than the continuous closure in repair of transverse incisions during the first 2 postoperative weeks. Both repair methods were associated with a similar maximum tensile strength at 4 postoperative weeks. Repair techniques using different bite sizes and intervals resulted in similar maximum tensile strengths.