Microbiology subsystem of a total, dedicated laboratory computer system.

The computer system used by the Microbiology Service of the Clinical Pathology Department, Clinical Center, National Institutes of Health is discussed. This microbiology subsystem is a part of a dedicated on-line laboratory computer system used by the entire department. The laboratory computer is connected on-line to a hospital computer which provides patient admission, transfer, and discharge data. Mark sense worksheets and cathode ray tube terminals are used for result entry and correction. Cumulative patient reports are printed. Results for both active and completed accessions can be easily retrieved on cathode ray terminals in the laboratory. All laboratory data are archived on magnetic tape from which a research data base and microfiched laboratory records are generated. The manner in which the system is integrated in the routine operation of the microbiology laboratory is emphasized. In addition, some of the costs, benefits, liabilities, and pitfalls associated with the introduction of the computer in the laboratory are reviewed. Finally, we have presented our concept of some of the future enhancements to our present system and some of the directions in which any future microbiology system might develop.     

AIDS-associated Kaposi''s sarcoma-derived cells in long-term culture express and synthesize smooth muscle alpha-actin.

Spindle-shaped cells from Kaposi's sarcoma lesions (AIDS-KS cells) were cultured for long periods in the presence of conditioned medium from activated CD4-positive T cells (HTLV-II infected transformed nonvirus producer) and characterized under in vitro conditions. To investigate a possible vascular origin, AIDS-KS cells were analyzed for the presence of smooth muscle alpha-actin, a differentiation marker for vascular smooth muscle cells. Immunofluorescence studies using a monoclonal antibody for smooth muscle alpha-actin demonstrated positive staining of the AIDS-KS cells (KS-3 and KS-4) but not by endothelial cells or fibroblasts. Northern blot analysis using an oligonucleotide probe unique for human smooth muscle alpha-actin indicated the expression of this gene by AIDS-KS cells. Similar analysis of biopsies from the KS lesion showed that in addition to the staining of smooth muscle cells associated with the blood vessels, the tumor-related spindle cells also stained positively. These cells were also analyzed for the expression of different growth factor genes. The platelet-derived growth factor (PDGF) A-chain gene was expressed at a moderate level. The insulin-like growth factor-1 (IGF-1) and insulin-like growth factor-2 (IGF-2) genes were not overexpressed in relation to control cells. These data suggest that the analyzed AIDS-KS cells may be smooth muscle-like cells and therefore of vascular origin. Based on these results as well as previous reports, we speculate that cells of the immune system may regulate growth of cells in the vascular wall by a novel pathway.     

B cell chronic lymphocytic leukaemia cells have reduced capacity to upregulate expression of MHC class I in response to interferon-gamma

AIMS: An important consideration in the design of a tumour vaccine is the ability of tumour-specific cytotoxic T lymphocytes (CTL) to recognise unmanipulated tumour cells in vivo. To determine whether B-CLL might use an escape strategy, the current studies compared B-CLL and normal B cell MHC class I expression. METHODS: Flow cytometry, TAP allele PCR and MHC class I PCR were used. RESULTS: While baseline expression of MHC class I did not differ, upregulation of MHC class I expression by B-CLL cells in response to IFN-gamma was reduced. No deletions or mutations of TAP 1 or 2 genes were detected. B-CLL cells upregulated TAP protein expression in response to IFN-gamma. Responsiveness of B-CLL MHC class I mRNA to IFN-gamma was not impaired. CONCLUSIONS: The data suggest that MHC class I molecules might be less stable at the cell surface in B-CLL than normal B cells, as a result of the described release of beta(2)m and beta(2)m-free class I heavy chains from the membrane. This relative MHC class I expression defect of B-CLL cells may reduce their susceptibility to CTL lysis in response to immunotherapeutic approaches.     

Construction and expression of recombinant plasmids encoding type 1 or D-mannose-resistant pili from a urinary tract infection Escherichia coli isolate.

Isolates of Escherichia coli from human urinary tract infections frequently express adherence properties found less often among normal intestinal isolates. These properties include adherence to human uroepithelial cells and primary monkey kidney cells, as well as D-mannose-resistant hemagglutination of human erythrocytes, and they are mediated by a pilus type different from type 1. The genes encoding this pilus type (pyelonephritis-associated pili, pap) and those encoding type 1 pili have been cloned from a urinary tract infection isolate of E. coli and transferred to an E. coli K-12 derivative. The recombinant plasmids were found to express functional pili and to endow the new host with all of the adherence properties of the urinary tract infection isolate. Both pilus types were found to be genetically distinct, and unlike the adherence genes from bovine, porcine, and human diarrheal isolates, both were found to be chromosomally encoded.     

Potential target sites in peripheral tissues for excitatory neurotransmission and excitotoxicity

Glutamate receptors (GluRs) are ubiquitously present in the central nervous system (CNS) as the major mediators of excitatory neurotransmission and excitotoxicity. Neural injury associated with trauma, stroke, epilepsy, and many neurodegenerative diseases such as Alzheimer's, Huntington's, and Parkinson's diseases and amyotrophic lateral sclerosis may be mediated by excessive activation of GluRs. Neurotoxicity associated with excitatory amino acids encountered in food, such as domoic acid and monosodium glutamate, has also been linked to GluRs. Less is known about GluRs outside the CNS. Recent observations suggest that several subtypes of GluRs are widely distributed in peripheral tissues. Using immunochemical and molecular techniques, the presence of GluR subtypes was demonstrated in the rat and monkey heart, with preferential distribution within the conducting system, nerve terminals, and cardiac ganglia. GluR subtypes NMDAR 1, GluR 2/3, and mGluR 2/3 are also present in kidney, liver, lung, spleen, and testis. Further investigations are needed to assess the role of these receptors in peripheral tissues and their importance in the toxicity of excitatory compounds. Therefore, food safety assessment and neurobiotechnology focusing on drugs designed to interact with GluRs should consider these tissues as potential target/effector sites.     

Exploring cancer register data to find risk factors for recurrence of breast cancer – application of Canonical Correlation Analysis

Background  

A common approach in exploring register data is to find relationships between outcomes and predictors by using multiple regression analysis (MRA). If there is more than one outcome variable, the analysis must then be repeated, and the results combined in some arbitrary fashion. In contrast, Canonical Correlation Analysis (CCA) has the ability to analyze multiple outcomes at the same time.     

The CTLA-4 gene region of chromosome 2q33 is linked to, and associated with, type 1 diabetes. Belgian Diabetes Registry

Susceptibility to autoimmune insulin-dependent (type 1) diabetes mellitus is determined by a combination of environmental and genetic factors, which include variation in MHC genes on chromosome 6p21 (IDDM1) and the insulin gene on chromosome 11p15 (IDDM2). However, linkage to IDDM1 and IDDM2 cannot explain the clustering of type 1 diabetes in families, and a role for other genes is inferred. In the present report we describe linkage and association of type 1 diabetes to the CTLA-4 gene (cytotoxic T lymphocyte associated-4) on chromosome 2q33 (designated IDDM12). CTLA-4 is a strong candidate gene for T cell- mediated autoimmune disease because it encodes a T cell receptor that mediates T cell apoptosis and is a vital negative regulator of T cell activation. In addition, we provide supporting evidence that CTLA-4 is associated with susceptibility to Graves' disease, another organ- specific autoimmune disease.      

Effect of defective connective tissue on the formation of aneurysmal-like structures in the rat testicular artery

Microscopic aneurysmal-like structures (ALS) develop spontaneously in the convoluted rat testicular artery and have been previously proposed as a model relevant to cerebral aneurysms. The effect of defects in connective tissue fibres on ALS formation was investigated by microscopy using two approaches: (i) the study of the effect of β-aminopropionitrile (BAPN), an inhibitor of the cross-linking of elastic and collagen fibres, on the incidence, size and morphology of ALS in spontaneously hypertensive rats (SHR) and their normotensive controls (WKY). The straight spermatic artery was studied for comparison. (ii) The determination of the incidence of spontaneous ALS in Brown Norway (BN) and Long Evans (LE) rats which are highly susceptible (BN) or resistant (LE) to the spontaneous rupture of the arterial internal elastic lamina. (i) BAPN increased the number and size of ALS in SHR and WKY rats and had no effect on the straight spermatic artery and (ii) ALS were more numerous and of greater size in BN than in LE rats. Taken together, these results show that defective connective tissue fibres may favour the formation and induce the enlargement of aneurysmal-like structures. By analogy, these data suggest that a lack of connective tissue fibre integrity may be of importance in cerebral aneurysm formation and development.