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An Sau3 AI restriction endonuclease isoschizomer from Bacillus cereus.

The isolation and characterization of a restriction endonuclease from Bacillus cereus IOC 243 are described. The enzyme recognizes the palindromic sequence 5'-G(met-A,A)TC-3' as determined by PEI chromatography of pancreatic DNase, snake venom phosphodiesterase digestion products of labelled fragments, analysis of restriction digests from normal and N6-methyladenine-free DNA and direct sequence analysis of cloned fragments. The staggered cleavage products with 5' -terminal pGATC extensions are efficiently labelled with polynucleotide kinase and are easily cloned into BamHI sites. The enzyme, denoted Bce243, is thus an isoschizomer of Sau3AI. Its use and potential advantages in substituting Sau3AI. are discussed.

Bacillus cereus↗

Expression of the human apolipoprotein AI gene fused to the E. coli gene for beta-galactosidase.

The human apoAI gene was expressed in E. coli by in-frame fusion to a modified beta-galactosidase gene present in plasmid pUR291. The fused beta-galactosidase-apoAI gene product was expressed at a high level and was recognized by an anti-human apoAI antiserum. Besides the fused protein, at least one degradation product having an Mr similar to that of beta-galactosidase was present in high amounts in bacterial extracts. These results and those of a pulse-chase experiment indicate that degradation took place only in the apoAI moiety of the chimeric protein.

Apolipoprotein A-I↗

Congenital tremor type AI: light and electron microscopical observations on the spinal cords of affected piglets.

The histology and ultrastructure of the spinal white matter from the dorsolateral funiculus of the third cervical segment was studied in normal control pigs and pigs whose dams were inoculated with the Weybridge congenital tremor strain of swine fever virus in early pregnancy. Only inoculated sows produced abnormal piglets. These showed congenital tremors and ataxia. The severity of clinical signs was related to the degree of spinal myelin deficiency. Morphologically this was quantified by determination of the thickness of myelin investing axons classed according to their diameter. In clinically affected pigs fewer axons were myelinated than normal. Though the myelin sheath thickness increased with increasing axon diameter in all pigs whether clinically normal or not, the increase was less in moderately affected and much less in severely affected pigs. The deficiency of spinal myelin was probably due to delayed or sub-normal myelination accompanied by paranodal myelin abnormalities, myelin degeneration and remyelination.

Animals↗

Expression and secretion of chicken apolipoprotein AI in transfected COS cells.

A full-length chicken apolipoprotein A-I (apoAI) cDNA has been cloned into an expression vector, pRSVapoAI. This plasmid was transfected into a monkey kidney (COS-1) cell line in order to study apolipoprotein-lipid assembly. Chicken apoAI is the major apolipoprotein of chicken high-density lipoprotein (HDL), which is less complex in apolipoprotein content than the HDL of human plasma. The transient transfected COS-1 cells synthesized and secreted authentic plasma apoAI. Under serum-free medium conditions, COS cells secreted only proapoAI. A small portion (15%) of the secreted apoAI floated at a density 1.07-1.20 g/ml. Upon incubation with fetal bovine serum at 10 degrees C, a majority of the apoAI was recovered in the HDL density (1.06-1.20 g/ml) region. Secreted apoAI was labeled when transfected COS cells were incubated with [U-14C]palmitate, but the incorporation of radioactivity was not the result of fatty acid acylation through ester bond formation. These results indicate that heterologous COS-1 cells are capable of synthesizing and secreting apoAI, and that intracellular association of apoAI with lipids is not necessary for secretion.

Acylation↗

SMR (simulating medical reasoning): an expert shell for non-AI experts.

SMR is an expert system shell designed to put the tools for knowledge acquisition directly into the hands of the domain expert. Since the knowledge base is represented as free text within a simplified syntactic structure, it is intelligible to anyone familiar with medical terminology. The knowledge base includes the rules for inference making as well as data groupings and protocols to facilitate case recording. In this paper, SMR is presented from the expert's point of view, describing the rule syntax and procedures for formulating the required diagnostic or therapeutic knowledge in the chosen domain. Similarly, the end-user's application of the system to patient data and the provisions for exploring and explaining the system's conclusions and reasoning processes are detailed. Avoiding tedious and often inane dialog, the user enters all that is known about the patient and receives a report of the system's conclusions and recommendations followed by a list of observations to be made in patient follow-up. An expert system for evaluating the diabetic patient is used to illustrate system operations.

Computer Simulation↗

Neisseria gonorrhoeae M.Ngo AI DNA methyltransferase: physical and catalytic properties of the homogeneous enzyme.

A DNA methyltransferase, M.NgoAI, was purified to homogeneity from Neisseria gonorrhoeae strain WR220 by successive column chromatography. Its Mr is 25,000, as determined by both gel filtration and denaturing polyacrylamide gel electrophoresis. Maximal enzymatic activity was obtained in 50 mM Tris.HCl (pH 7.4), 10 mM EDTA, with incubation at 37 degrees C. An apparent Km value for S-adenosylmethionine and 5' -GGCC sites was determined to be 1.25 microM and 89.6 nM, respectively.

Bacterial Proteins↗

Isolation, characterization and sequencing of the chicken apolipoprotein-AI-encoding gene.

The chicken ApoAI gene has been isolated and characterized. This gene contains three introns: the first is situated after nucleotide (nt) 41 in the 5'-untranslated region of the gene, the second interrupts the codon specifying the Gly-10 of the prepeptide, and the third disrupts the codon for Asp42 in the mature ApoAI protein. The chicken ApoAI gene has 62.6% sequence similarity with the human and 65.0% similarity with the rat gene. Intron-exon organization of the chicken gene is similar to that of human and rat ApoAI genes. Two different transcriptional start points (tsp), only 2 nt apart from each other, have been obtained for the chicken ApoAI gene. The 5'-flanking sequence of the gene contains TAAATA (TATA-like box) and CCACAT (CCAAT-like box) sequences, which are located 21 bp and 96 bp upstream from the tsp, respectively. This gene's sequence and structural organization provide a basis for future studies of the regulation of chicken ApoAI gene expression.

Amino Acid Sequence↗

The adolescence of AI in medicine: will the field come of age in the '90s?

Artificial intelligence in medicine (AIM) has reached a period of adolescence in which interactions with the outside world are not only natural but mandatory. Although the basic research topics in AIM may be those of artificial intelligence, the applied issues touch more generally on the broad field of medical informatics. To the extent that AIM research is driven by performance goals for biomedicine, AIM is simply one component within a wide range of research and development activities. Furthermore, an adequate appraisal of AIM research requires an understanding of the research motivations, the complexity of the problems, and a suitable definition of the criteria for judging the field's success. Effective fielding of AIM systems will be dependent on the development of integrated environments for communication and computing that allow merging of knowledge-based tools with other patient data-management and information-retrieval applications. The creation of this kind of infrastructure will require vision and resources from leaders who realize that the practice of medicine is inherently an information-management task and that biomedicine must make the same kind of coordinated commitment to computing technologies as have other segments of our society in which the importance of information management is well understood.

Artificial Intelligence↗