Light and electron microscopic analysis of two divisions of the suprachiasmatic nucleus in the young and aged rat.

The suprachiasmatic nucleus (SCN) is a principal controller of mammalian circadian rhythms. However, in spite of documented disturbance of biological rhythms in old animals, few significant age-related changes have been observed in this nucleus. This study examined age-related differences in SCN volume, neuronal number, density, and ultrastructural features in the entire rat SCN and in its two divisions, the denser ventromedial (compacta) and less dense dorsolateral (dissipata). Light and electron microscopic morphometric techniques were utilized in weanlings (21-28 days), young adults (3-6 mo), and aged (30-36 mo) animals. The total SCN volume, as well as volumes of the compacta region, were significantly greater in young adult and aged rats than in weanlings. Thus, as the rat ages the SCN increases in total size. However, the dissipata region appears to decrease in volume while the compacta increases. Even though the total number of SCN neurons was quite constant in the three age groups, the number of neurons in the dissipata region was decreased significantly in the young adult and aged groups as compared to the weanling. Neurons in the compacta region were usually spindled-shaped with two dendritic processes, while oval to spheroidal cells with 3-4 processes predominated in the dissipata. Nuclei of SCN cells were often invaginated. In weanlings, more SCN neuronal nuclei had invaginated nuclei in the dissipata region (66%) compared to the compacta (37%). In the two older age groups of rats, a higher percentage of invaginated neuronal nuclei were found in both regions. However, more were still found in the dissipata (90%) compared to the compacta (72%), even though the number of these cells in the compacta doubled. Thus, there was a large increase in the number of invaginated nuclei, as well as the number of invaginations, in the young adult rats compared to the weanling group, and this increase persisted in aged rats. SCN neurons usually had nuclei surrounded by a thin perimeter of cytoplasm containing sparse mitochondria and granular endoplasmic reticulum, multiple Golgi regions, and a moderate number of free ribosomes. In weanlings, mitochondria contained dense cristae and the granular endoplasmic reticulum was relatively prominent. Degenerative ultrastructural changes which included mitochondrial enlargement/vacuolation, Golgi vacuolation, lysosome, and lipofuscin development occurred in less than 10% of young adult SCN cells, and were more frequently found in the dissipata. In aged, rats 30% of the neurons showed degenerative changes in the dissipata compared with 18% in the compacta. Degenerative changes appeared highly correlated with the degree of membrane folding.(ABSTRACT TRUNCATED AT 400 WORDS)

Recognition by the human immune system of candidate vaccine epitopes of Toxoplasma gondii measured by a competitive ELISA.

A competitive ELISA for the detection of human antibody to specific candidate vaccine epitopes of Toxoplasma gondii is described. The method is based on competition between antibody in human serum and murine monoclonal antibody to bind to a crude sonicated antigen of T. gondii. A total of 301 normal human sera were tested against a panel of four anti-T. gondii murine monoclonal antibodies. The results obtained with two of the monoclonal antibodies (FMC 22 and FMC 23) gave good agreement with those of an indirect ELISA for human IgG to T. gondii, a measure of previous exposure to the parasite. The epitopes against which these two monoclonal antibodies are directed were recognized by the immune system of all patients previously exposed to the parasite, while greater than 92 and greater than 96% respectively of these patients recognized the epitopes which the other two monoclonal antibodies (FMC 18 and FMC 19) are directed against. An ELISA double antibody binding system was used to ascertain relationships in the epitope binding sites of the four monoclonal antibodies. The data obtained demonstrated that FMC 19 and FMC 22 bound to related epitopes. FMC 18, FMC 22 and FMC 23 also bound to related epitopes, but the binding of FMC 19 was found to be independent of the binding of FMC 18 and FMC 23 to their epitope binding sites.

Serotyping of Campylobacter jejuni from an outbreak of enteritis implicating chicken.

An outbreak of campylobacter enteritis involving 7 of 17 people over a period of 5 days followed a dinner at a restaurant. A chicken casserole dish was implicated with a food-specific attack rate of 58%. Campylobacter jejuni Penner serotype 18/21/29, resistant to metronidazole, was isolated from 3 of 4 symptomatic patients and from three raw fresh chicken samples closely associated with the implicated chicken. Numbers of C. jejuni in the chicken ranged from 5.3 X 10(1) to 7.5 X 10(2) colony forming units per square centimeter of surface area. This is the first outbreak of campylobacter enteritis reported in Australia in which C. jejuni has been isolated from both human and food sources and the isolates serologically confirmed as identical.

Aeromonas hydrophila typing scheme based on patterns of agglutination with erythrocytes and yeast cells.

An agglutination typing scheme has been developed for strains of Aeromonas hydrophila. Primary agglutination typing is based on testing agar-grown A. hydrophila cells with human, horse, rat, and guinea pig erythrocytes and Saccharomyces cerevisiae cells. Further subdivision of primary groups is based firstly on whether yeast cell agglutination is inhibited by a D-mannose polymer, yeast mannan, and secondly on patterns of inhibition of hemagglutination by yeast mannan and the monomeric sugars L-fucose, D-galactose, and D-mannose. A total of 320 isolates were tested, and these were divisible into 39 distinct types on the basis of this scheme. Application of this typing scheme in the future to isolates of A. hydrophila known to be associated with human infection may enable correlations to be made between particular agglutination types and human pathogenicity.

Coliphage 186 Replication is delayed when the host cell is UV irradiated before infection.

In contrast to results with injections by lambda and P2, the latent period for infection by coliphage 186 is extended when the host cell is UV irradiated before infection. We find that 186 replication is significantly delayed in such a cell, even though the phage itself has not been irradiated. In contrast, replication of the closely related phage P2 under the same conditions is not affected.

The transient inability of the conjugating female cell to host 186 infection explains the absence of zygotic induction for 186.

In an Hfr(186) X F- cross, the 186 prophage on the incoming male chromosome is not induced, despite the fact that prophage 186 can be induced by other means (W. H. Woods and J.B. Egan, J Virol. 14:1349-1356, 1974). We show here that the conjugating female is temporarily inhibitory to infection by 186, and this delay, we postulate, enables cI repression to be reestablished before the female cell recovers its 186 sensitivity.

Prophage induction of noninducible coliphage 186.

Coliphage 186 has been regarded as a member of the noninducible group of coliphages. Evidence that prophage 186 is induced by ultraviolet irradiation or by treatment with nalidixic acid or mitomycin C is now presented. The phage yields were similar to those from lysogens of the inducible phage lambda, and the induction required a recA(+) host. A noninducible mutant of 186 was isolated from its heat-inducible derivative, 186cIts, that was no longer inducible by ultraviolet irradiation but remained heat inducible. That zygotic induction of 186 after transfer from a lysogenic male to a non-lysogenic recipient did not occur is indicated by the following findings: (i) there was only a slight increase in phage titer; (ii) similar levels of recombinants were obtained for markers adjacent or distal to the phage integration site, whether the recipient was lysogenic or not, and there was no effect on the gradient of marker transfer; (iii) lysogenic recombinants were readily found and the co-transfer of 186 with adjacent markers was the same to lysogenic or non-lysogenic recipients. Thus, 186 formed an inducible prophage that did not display zygotic induction. Nevertheless, it shared many properties with the noninducible phage P2 as outlined in the discussion.

Integration stie of noninducible coliphage 186.

From conjugational data, the attachment site for noninducible coliphage 186 (att186) was located between the origins of Hfr strains KL16 and KL98, and close to the pheA gene in Escherichia coli K-12. P1 transductions indicated that att186 lies at 51 min on the standard genetic map of E. coli, with the order cysC-nalB-att186-pheA. The presence of prophage 186 in the donor destroyed linkage between nalB and pheA, which is taken as evidence for the integration of the 186 prophage between these genes.