An overview of some mechanisms of bacterial pathogenesis

The adherence of microorganisms to host surfaces is highly specific, and in many cases, essential for subsequent pathogenetic events to occur. A dynamic process leading to increased mucosal adherence of gram-negative bacilli to epithelial cell receptors in the oral cavity appears to be the initial step in the development of pneumonia. In infectious processes secondary to Streptococcus pneumoniae, adherence may also play a role in specific syndromes. In many cases, however, colonization of oropharyngeal mucus itself, the presence of capsular polysaccharide, and the release of various cell wall components appear to interact to cause clinical disease. In Neisseria gonorrhoeae infections, adherence is all important and is mediated by a number of cell surface structures. These have been studied extensively. Many of these structures, such as pili and protein II, exhibit great variability both between strains and in the same organism at different stages of infection. Others, such as protein I, are more constant. This information has been used in the production of specific vaccines to more preserved structures to inhibit adherence. These will be tested in the near future. It is our view that a better understanding of the many forms of bacterial adherence will be the key to our designing more effective strategies to detect early infection and to intervene more decisively to limit its spread.     

A social learning model of conflict and cooperation in human societies

Patterns of conflict and cooperation both within and between societies may be related to the degree of cultural similarity within and between the same societies. A simple model of social learning is used to predict patterns of conflict and cooperation in hypothetical societies that differ in the roles of relatives and nonrelatives in the enculturation of children. The model is illustrated by comparing its predictions to known differences in the patterns of conflict between males inpatrilocal and matrilocal societies.     

Sequence heterogeneity and anomalous electrophoretic mobility of kinetoplast minicircle DNA from Leishmania tarentolae

Several unit-length minicircles from the kinetoplast DNA of Leishmania tarentolae were cloned into pBR322 and into M13 phage vectors. The complete nucleotide sequences of three different partially homologous minicircles were obtained. The molecules contained a region of approx. 80% sequence homology extending for 160–270 bp and a region unique to each minicircle. A 14-mer was found to be conserved in all kinetoplast minicircle sequences reported to date. The frequency distributions of various minicircle sequence classes in L. tarentolae were obtained by quantitative gel electrophoresis and by examination of the “T ladder” patterns of minicircles randomly cloned into M13 at several sites. By these methods we could assign approx. 50% of the total minicircle DNA into a minimum of five sequence classes. A sequence-dependent polyacrylamide gel migration abnormality was observed with several minicircle fragments both cloned and uncloned. The abnormality was dependent on the presence of a portion of the conserved region of the minicircle.     

Spontaneous Aggregation and Cytotoxicity of the β-Amyloid Aβ1–40: A Kinetic Model

The time dependency of the spontaneous aggregation of the fibrillogenic β-Amyloid peptide, Aβ^1–40, was measured by turbidity, circular dichroism, HPLC, and fluorescence polarization. The results by all methods were comparable and they were most consistent with a kinetic model where the peptide first slowly forms an activated monomeric derivative (AM), which is the only species able to initiate, by tetramerization, the formation of linear aggregates. The anti-Aβ antibody 6E10, raised against residues 1–17, at concentrations of 200–300 nM delayed significantly the aggregation of 50 μM amyloid peptide. The anti–Aβ antibody 4G8, raised against residues 17–24, was much less active in that respect, while the antibody A162, raised against the C-terminal residues 39–43 of the full-length Aβ was totally inactive at those concentrations. Concomitant with the aggregation experiments, we also measured the time dependency of the Aβ^1–40–induced toxicity toward SH-EP1 cells and hippocampal neurons, evaluated by SYTOX Green fluorescence, lactate dehydrogenase release, and activation of caspases. The extent of cell damage measured by all methods reached a maximum at the same time and this maximum coincided with that of the concentration of AM. According to the kinetic scheme, the latter is the only transient peptide species whose concentration passes through a maximum. Thus, it appears that the toxic species of Aβ^1–40 is most likely the same transient activated monomer that is responsible for the nucleation of fibril formation. These conclusions should provide a structural basis for understanding the toxicity of Aβ^1–40 in vitro and possibly in vivo.     

A simple,rapid, and sensitive method for measuring protein concentration in subcellular membrane fractions prepared by sucrose density ultracentrifugation

The Multiskan spectrophotometric system and Coomassie brilliant blue G-250 protein-dye binding method have been used together to measure NaOH-solubilized protein in subcellular membrane fractions prepared from isolated rat adipose cells. Forty-eight samples can be read in duplicate within 1 min. Sucrose in concentrations up to 0.7 m interfere only moderately with the assay. A rapid and convenient method is, therefore, now available for multiple protein determinations following sucellular fractionation on sucrose gradients.     

A radioimmunoassay for epinephrine and norepinephrine in tissues and plasma

An I¹²⁵ radioimmunoassay (RIA) has been developed for the measurement of plasma and tissue epinephrine (E) and norepinephrine (NE). The assay utilizes an antibody which specifically binds metanephrine. E and NE are detected by conversion to metanephrine with the enzymes catechol-0-methyltransferase and phenylethanol-amine-N-methyltransferase. The assay is very specific and will allow the measurement of E and NE in less than 500 μl of normal human plasma. E and NE concentrations were determined by both the RIA and a radioenzymatic assay in canine, human and rat biologic samples. The correlation coefficients between the two assays were .962 for E and .956 for NE. The RIA is sensitive, specific, precise and significantly less costly and time consuming than present radioenzymatic methods.