A systematic, genome-wide, phenotype-driven mutagenesis programme for gene function studies in the mouse.

As the human genome project approaches completion, the challenge for mammalian geneticists is to develop approaches for the systematic determination of mammalian gene function. Mouse mutagenesis will be a key element of studies of gene function. Phenotype-driven approaches using the chemical mutagen ethylnitrosourea (ENU) represent a potentially efficient route for the generation of large numbers of mutant mice that can be screened for novel phenotypes. The advantage of this approach is that, in assessing gene function, no a priori assumptions are made about the genes involved in any pathway. Phenotype-driven mutagenesis is thus an effective method for the identification of novel genes and pathways. We have undertaken a genome-wide, phenotype-driven screen for dominant mutations in the mouse. We generated and screened over 26,000 mice, and recovered some 500 new mouse mutants. Our work, along with the programme reported in the accompanying paper, has led to a substantial increase in the mouse mutant resource and represents a first step towards systematic studies of gene function in mammalian genetics.

A radiation hybrid map of mouse genes.

A comprehensive gene-based map of a genome is a powerful tool for genetic studies and is especially useful for the positional cloning and positional candidate approaches. The availability of gene maps for multiple organisms provides the foundation for detailed conserved-orthology maps showing the correspondence between conserved genomic segments. These maps make it possible to use cross-species information in gene hunts and shed light on the evolutionary forces that shape the genome. Here we report a radiation hybrid map of mouse genes, a combined project of the Whitehead Institute/Massachusetts Institute of Technology Center for Genome Research, the Medical Research Council UK Mouse Genome Centre, and the National Center for Biotechnology Information. The map contains 11,109 genes, screened against the T31 RH panel and positioned relative to a reference map containing 2,280 mouse genetic markers. It includes 3,658 genes homologous to the human genome sequence and provides a framework for overlaying the human genome sequence to the mouse and for sequencing the mouse genome.

A pilot clinical trial of two murine monoclonal antibodies fixing human complement in patients with chronic lymphatic leukaemia.

The use of monoclonal antibodies (MABs) for the therapy of malignant diseases offers the potential advantage of greater target cell specificity, and therefore less toxicity. A major limitation of this therapeutic approach has been the inability of most MABs to kill the cell once bound to the target antigen. We have previously reported the development of two murine IgM MABs, WM63 (CD48) and WM66 (unclustered), that react with panleucocyte antigens widely expressed on cells from lymphoproliferative disorders, and are lytic with human complement. These antibodies have subsequently been administered intravenously to patients with chronic lymphocytic leukaemia (CLL) in a Phase One trial. Seven patients with progressive CLL received increasing daily doses of WM66 (Patients 1-3) or WM63 (Patients 4-7), with one patient also receiving a continuous infusion of WM63 over 20 hours. All patients demonstrated a significant but transient reduction in the number of circulating leucocytes, and no overall effect on disease progression was observed. Antibody coating of circulating lymphocytes was seen in patients receiving WM-63. Patients receiving large doses of WM63 (cases 5-7) demonstrated a decline in complement levels during treatment. There were no major adverse reactions to WM66, but two patients developed dose limiting side effects to WM63. No human anti-mouse antibody (HAMA) responses were documented. These findings indicate that in vitro cytotoxicity mediated by Mabs fixing human complement correlates poorly with clinical responses, and support earlier observations which indicate that cell-mediated cytotoxicity is necessary for effective antibody therapy.

Coping with anxiety and panic: a factor analytic study.

OBJECTIVES: This study aimed to explore the coping styles that agoraphobia sufferers adopt when attempting to cope with symptoms of anxiety and panic. It aimed to extend Watts's (1989) Coping with Anxiety Questionnaire (CAQ) by including items to assess self-vigilance. It was hypothesized that agoraphobia sufferers would adopt consistent coping styles that would be related to symptom severity. DESIGN: A factor analysis was performed on questionnaire data. METHOD: A postal questionnaire was completed by members (N = 112) of a self-help group for agoraphobia and panic sufferers. All participants completed the Beck Anxiety Inventory, an Agoraphobia Severity Scale and a slightly modified version of the CAQ. Coping styles were identified via factor analysis of the CAQ items. RESULTS: Three coping styles were identified, which were labelled Effective Coping, Avoidant Coping and Self-vigilance. The latter two coping styles were found to be correlated with increased levels of agoraphobic symptomatology and with higher levels of anxiety. CONCLUSIONS: The present results support the previous research on coping tactics in anxiety and are compatible with cognitive therapy accounts of the role of self-vigilance in anxiety disorders.

Unusual presentation of granulocytic sarcoma in the breast: a case report and review of the literature.

This is a case of granulocytic sarcoma presenting as bilateral breast masses in a 40-yr-old woman with concurrent unsuspected chronic myeloid leukemia diagnosed by fine-needle aspiration. The granulocytic differentiation was recognized on Diff-Quik-stained cytology smears and confirmed rapidly on flow cytometry on the same day. The breast has been reported to be an uncommon site for granulocytic sarcoma. We found that 38.8% of granulocytic sarcomas diagnosed by fine-needle aspiration in the English-language literature occurred in the breast. In the absence of clinical history or hematological abnormality, granulocytic sarcoma may be misdiagnosed, depending on the degree of myeloid differentiation present within the tumor. The differential diagnosis includes large-cell non-Hodgkin's lymphoma, lobular carcinoma of the breast, undifferentiated carcinoma, malignant melanoma, extramedullary hemopoiesis and inflammation. The key morphological features and useful ancillary tests are discussed.

A novel cytogenetic abnormality in Burkitt lymphoma associated with treatment resistant disease.

We present a patient with deletion of IgH associated with the reciprocal translocation (8;14) in Burkitt lymphoma. The patient had treatment resistant disease and died 10 weeks after diagnosis. The deletion was detected by fluorescence in situ hybridization at diagnosis and again after failure of chemotherapy. To our knowledge this is the first report of such a deletion.

Geriatric medicine in a community hospital.

A study is reported of the care that was being received by the patients over the age of 60 years in general practitioner community hospital beds at Abingdon Hospital in Berkshire. The appropriateness of this care is discussed and measures are suggested for improving it. The practical working relationship between general practitioners attending the community hospital and the local consultant physicians in geriatric medicine is also discussed and again measures for improving their combined services to the community hospital are suggested.

Isolation and characterization of 2,3-dichloro-1-propanol-degrading rhizobia.

2,3-Dichloro-1-propanol is more chemically stable than its isomer, 1, 3-dichloro-2-propanol, and is therefore more difficult to degrade. The isolation of bacteria capable of complete mineralization of 2, 3-dichloro-1-propanol was successful only from enrichments at high pH. The bacteria thus isolated were found to be members of the alpha division of the Proteobacteria in the Rhizobium subdivision, most likely Agrobacterium sp. They could utilize both dihaloalcohol substrates and 2-chloropropionic acid. The growth of these strains in the presence of 2,3-dichloro-1-propanol was strongly affected by the pH and buffer strength of the medium. Under certain conditions, a ladder of four active dehalogenase bands could be visualized from this strain in activity gels. The enzyme involved in the complete mineralization of 2,3-dichloro-1-propanol was shown to have a native molecular weight of 114,000 and consisted of four subunits of similar molecular weights.

Ontologies for the description of mouse phenotypes.

Ontologies are becoming increasingly important for the efficient storage, retrieval and mining of biological data. The description of phenotypes using ontologies is a particularly complex problem. We outline a schema that can be used to describe phenotypes by combining orthologous axiomatic ontologies. We also describe tools for storing, browsing and searching such complex ontologies. Central to this approach is that assays (protocols for measuring phenotypic characters) describe what has been measured as well as how this was done, allowing assays to link individual organisms to ontologies describing phenotypes. We have evaluated this approach by automatically annotating data on 600,000 mutant mice phenotypes using the SHIRPA protocol. We believe this approach will enable the flexible, extensible and detailed description of phenotypes from any organism.

Informatics for mutagenesis: the design of mutabase--a distributed data recording system for animal husbandry, mutagenesis, and phenotypic analysis.

The increasing use of high-throughput methods for the production of biologically important information and the increasing diversity of that information pose considerable bioinformatics challenges. These challenges will be met by implementing electronic data management systems not only to capture the data, but increasingly to provide a platform for data integration and mining as we enter the post-genomic era. We discuss the design and implementation of such a data capture system, 'Mutabase', as a model of how such electronic systems might be designed and implemented. Mutabase was created in support of a large-scale, phenotype-driven mouse mutagenesis program at MRC Mammalian Genetics Unit, Harwell, in collaboration with SmithKline Beecham Pharmaceuticals, Queen Mary and Westfield College, London, and Imperial College of Science, Technology and Medicine, London. The aim of this mutagenesis project is to make a significant contribution to the existing mouse mutant resource, closing the phenotype gap and providing many more models for fundamental research and disease modeling. Mutabase records experimental details at the 'point of generation' and provides a number of dissemination and analysis tools for the experimental data, as well as providing a means of assessing various aspects of progress of the program. Mutabase uses a hypertext-based interface to provide interaction between a number of intranet-based client workstations and a central industrial strength database. Mutabase utilizes a variety of techniques in order to implement the user interface system including Perl/CGI, Java Servlets, and an experimental CORBA server. We discuss the relative merits of these methods in the context of the need to provide sound informatics approaches for the support of systematic mutagenesis programs.