Microarray and proteomic analysis of the human alcoholic brain

The superior frontal cortex (SFC) is selectively damaged in chronic alcohol abuse, with localized neuronal loss and tissue atrophy. Regions such as motor cortex show little neuronal loss except in severe co-morbidity (liver cirrhosis or WKS). Altered gene expression was found in microarray comparisons of alcoholic and control SFC samples [1]. We used Western blots and proteomic analysis to identify the proteins that also show differential expression. Tissue was obtained at autopsy under informed, written consent from uncomplicated alcoholics and age- and sex-matched controls. Alcoholics had consumed 80 g ethanol/day chronically (often, 200 g/day for 20 y). Controls either abstained or were social drinkers ( 20 g/day). All subjects had pathological confirmation of liver and brain diagnosis; none had been polydrug abusers. Samples were homogenized in water and clarified by brief centrifugation (1000g, 3 min) before storage at –80°C. For proteomics the thawed suspensions were centrifuged (15000g, 50 min) to prepare soluble fractions. Aliquots were pooled from SFC samples from the 5 chronic alcoholics and 5 matched controls used in the previous microarray study [1]. 2-Dimensional electrophoresis was performed in triplicate using 18 cm format pH 4–7 and pH 6–11 immobilized pH gradients for firstdimension isoelectric focusing. Following second-dimension SDS-PAGE the proteins were fluorescently stained and the images collected by densitometry. 182 proteins differed by 2-fold between cases and controls. 141 showed lower expression in alcoholics, 33 higher, and 8 were new or had disappeared. To date 63 proteins have been identified using MALDI-MS and MS-MS. Western blots were performed on uncentrifuged individual samples from 76 subjects (controls, uncomplicated alcoholics and cirrhotic alcoholics). A common standard was run on every gel. After transfer, immunolabeling, and densitometry, the intensities of the unknown bands were compared to those of the standards. We focused on proteins from transcripts that showed clear differences in a series of microarray studies, classified into common sets including Regulators of G-protein Signaling and Myelin-associated proteins. The preponderantly lower level of differentially expressed proteins in alcoholics parallels the microarray mRNA analysis in the same samples. We found that mRNA and protein expression do not frequently correspond; this may help identify pathogenic processes acting at the level of transcription, translation, or post-translationally.

Using integrated metamodeling to define OO design patterns with object-z and UML

Three important goals in describing software design patterns are: generality, precision, and understandability. To address these goals, this paper presents an integrated approach to specifying patterns using Object-Z and UML. To achieve the generality goal, we adopt a role-based metamodeling approach to define patterns. With this approach, each pattern is defined as a pattern role model. To achieve precision, we formalize role concepts using Object-Z (a role metamodel) and use these concepts to define patterns (pattern role models). To achieve understandability, we represent the role metamodel and pattern role models visually using UML. Our pattern role models provide a precise basis for pattern-based model transformations or refactoring approaches.

A case study in specification and implementation testing

Achieving consistency between a specification and its implementation is an important part of software development In previous work, we have presented a method and tool support for testing a formal specification using animation and then verifying an implementation of that specification. The method is based on a testgraph, which provides a partial model of the application under test. The testgraph is used in combination with an animator to generate test sequences for testing the formal specification. The same testgraph is used during testing to execute those same sequences on the implementation and to ensure that the implementation conforms to the specification. So far, the method and its tool support have been applied to software components that can be accessed through an application programmer interface (API). In this paper, we use an industrially-based case study to discuss the problems associated with applying the method to a software system with a graphical user interface (GUI). In particular, the lack of a standardised interface, as well as controllability and observability problems, make it difficult to automate the testing of the implementation. The method can still be applied, but the amount of testing that can be carried on the implementation is limited by the manual effort involved.

Testing java interrupts and timed waits

Testing concurrent software is difficult due to problems with inherent nondeterminism. In previous work, we have presented a method and tool support for the testing of concurrent Java components. In this paper, we extend that work by presenting and discussing techniques for testing Java thread interrupts and timed waits. Testing thread interrupts is important because every Java component that calls wait must have code dealing with these interrupts. For a component that uses interrupts and timed waits to provide its basic functionality, the ability to test these features is clearly even more important. We discuss the application of the techniques and tool support to one such component, which is a nontrivial implementation of the readers-writers problem.

Bridging technical and HCI research: Creating usable ubiquitous computing

This paper describes methods used to support collaboration and communication between practitioners, designers and engineers when designing ubiquitous computing systems. We tested methods such as “Wizard of Oz” and design games in a real domain, the dental surgery, in an attempt to create a system that is: affordable; minimally disruptive of the natural flow of work; and improves human-computer interaction. In doing so we found that such activities allowed the practitioners to be on a ‘level playing ground’ with designers and engineers. The findings we present suggest that dentists are willing to engage in detailed exploration and constructive critique of technical design possibilities if the design ideas and prototypes are presented in the context of their work practice and are of a resolution and relevance that allow them to jointly explore and question with the design time. This paper is an extension of a short paper submitted to the Participatory Design Conference, 2004.

Modelling and using imperfect context information

Most recently developed context-aware software applicationsmake unrealistic assumptions about the quality of the available context information, which can lead to inappropriate actions by the application and frustration on the part of the user. In this paper, we explore the problem of imperfect context information and some of its causes, and propose a novel approach for modelling incomplete and inaccurate information. Additionally, we present a discussion of our experiences in developing a contextaware communication application, highlighting design issues that are pertinent when developing applications that rely on imperfect context information.