Injury-induced NF-kappaB activation in the hippocampus: implications for neuronal survival.

Nuclear factor (NF)-kappaB p50 protein is involved in promoting survival in hippocampal neurons after trimethyltin (TMT)-injury. In the current study, hippocampal NF-kappaB activity was examined and quantitated from transgenic kappaB-lacZ reporter mice after chemical-induced injury. NF-kappaB activity was localized primarily to hippocampal neurons and significantly elevated over that in saline-treated mice between 4 and 21 days after TMT injection. Seven days after TMT injection, a timepoint of elevated NF-kappaB activity, gene expression in the hippocampus was studied by microarray analysis through comparison of expression profiles between treated nontransgenic and p50-null mice with their saline-injected controls. Seventeen genes increased in nontransgenic TMT-treated mice relative to saline-treated as well as showing no increase in p50-null mice, indicating a role for p50 in their regulation. One of these genes, the Na+, K+-ATPase-gamma subunit, was detected in brain for the first time. Several of the genes modulated by NF-kappaB are potentially related to neuroplasticity, providing additional evidence that this transcription factor is a neuroprotective signal in the hippocampus.

Mice expressing human mutant presenilin-1 exhibit decreased activation of NF-kappaB p50 in hippocampal neurons after injury.

Mutations in the presenilin-1 (mutPS-1) gene, a cause of familial Alzheimer's disease, increase the susceptibility of neurons to apoptotic death. Using the trimethyltin model of hippocampal neurodegeneration, mice expressing the human mutPS-1 gene (M146L) exhibited increased neurodegeneration and mortality relative to non-transgenic littermates. Activation of NF-kappaB p50 was found to be impaired in transgenic mice with unaltered expression levels suggesting that mutPS-1 expression inhibits p50 activation to adversely affect neuronal resistance to injury.

Signal transduction and neurosurvival in experimental models of brain injury.

Brain injury and neurodegenerative disease are linked by their primary pathological consequence-death of neurons. Current approaches for the treatment of neurodegeneration are limited. In this review, we discuss animal models of human brain injury and molecular biological data that have been obtained from their analysis. In particular, signal transduction pathways that are associated with neurosurvival following injury to the brain are presented and discussed.

Hypomethylation of cytosine 5-methyltransferase in human neoplasms.

Cytosine methylation is an epigenetic modification of DNA involved in control of gene expression. Neoplastic cells exhibit various alterations both in DNA methylation and activity of the enzyme responsible for this modification, 5-methyltransferase (5-MeTase). As there is little requirement for 5-methyltransferase expression in normal cells except during mitosis, we argued that the gene would be hypermethylated in normal cells. Southern analysis revealed almost complete methylation of the gene in genomic DNA from the peripheral blood leukocytes of healthy subjects and a primary fibroblast derived cell line. In contrast, in DNA from a range of tumour tissues and tumour derived cell lines, 5-MeTase exhibited marked hypomethylation. The results of this study indicate that dysregulation of the DNA methylating machinery, especially with respect to the methylation status of 5-MeTase, is a feature of a wide range of neoplasms.

An in vitro study of the effects of venom of Australian elapids on murine skeletal muscle and the protective effect of homologous plasma

The venom of many dangerous Australian snakes has a myotoxic component and some are strongly myolytic. The myotoxicity of venom of seven Australian elapid snakes was studied to determine their relative in vitro potency in causing cell death of C2C12 cells, a myoblast cell line, and murine pmyotubes in mixed cell culture. The venom of Pseudechis australis proved to be most myotoxic, Austrelaps superbus and Pseudechis porphyriacus venoms also exhibited myotoxicity relative to the other venoms tested. The specificity of Pseudechis porphyriacus venom was tested using the human glioma cell line TC3 and was shown to exhibit a general cytotoxicity. Myotoxicity, however, was the predominant action of the venom. It has long been known that certain animals sucha as the mongoose (herpestes edwardsii) are able to survive envenomation. Some species of snakes also possess this property and the neutralising factor(s) responsible for this P. porphyriacus has been shown to be present in the serum. The protective effect of homologous plasma from P. porphyriacus venom was also studied with reference to myotoxicity and cytotoxicity. The results of this study clearly demonstrated protection by homologous plasma using a myoblast cell line, C2C12, a primary mixed cell culture and TC3 cells. While protection was clear, particularly using high concentrations of venom, it was not absolute, and homologous plasma did not afford continued protection from the effects of the venom. In the mixed culture experiments venom/plasma mixtures pre-incubated for 30 min were more protective than venom/plasma mixtures which were not incubated, in contrast to the results of cell culture studies, which showed little difference.

Resident and infiltrating immune cells in the uveal tract in the early and late stages of experimental autoimmune uveoretinitis.

PURPOSE: To investigate the dynamics of resident and infiltrating immune cells in the choroid and iris during the early and late stages of experimental autoimmune uveitis (EAU) in Lewis rats. METHODS: Uveoretinitis was induced by footpad injection of crude retinal extract and complete Freund's adjuvant with concurrent intraperitoneal injection of Bordetella pertussis. Five experimental (EAU) and five control animals (adjuvant alone) were studied at days 5, 7, 9, 11 (prodromal stage) and 42 (late stage) after immunization. Five normal animals and five animals injected with B. pertussis alone served as further controls. Immunohistochemical localization of resident macrophages, major histocompatibility complex class II (Ia)+ dendritic cells (DC), infiltrating mononuclear cells, and T cells was performed on wholemounts of isolated choroidal and iris tissue. RESULTS: Double immunolabeling confirmed the presence of distinct networks of macrophages (591 +/- 52 cells/mm2) and DC (746 +/- 38 cells/mm2) in the rat choroid. No marked qualitative and quantitative changes were observed in the density or morphologic appearance of ED2+ resident tissue macrophages in the choroid and iris before clinical onset of ocular disease. On day 11, infiltration of ED1+ monocytes had occurred in the iris but not in the choroid; however, marked infiltration of T cells was evident in both choroid (286 +/- 161 cells/mm2) and iris (196 +/- 72 cells/mm2). The total density of Ia+ cells was significantly elevated in the choroid (1152 +/- 192 cells/mm2) at day 11, and small, round Ia+ cells were two to three times more frequent than normal at both sites. The density of T cells and Ia+ cells remained significantly elevated in the choroid and iris in the late stages of EAU. CONCLUSIONS: These data suggest resident uveal tract macrophages undergo no significant alteration in density in the early stages of EAU and that the earliest site of mononuclear cellular infiltrate in EAU occurs in the iris. The increased total density of Ia+ cells in the choroid on day 11 and the presence of significantly increased numbers of small, round Ia+ cells in the iris and choroid may represent increased trafficking of DC in the eye during uveoretinitis. Furthermore, the raised numbers of Ia+ cells, concurrent with the influx of T cells, suggests Ia+ DC and macrophages may act as local antigen-presenting cells in the induction of uveoretinitis.

Cervical cytology in Western Australia. Frequency, geographical and socioeconomic distributions and providers of the service.

Basic data were obtained from the records of 16,069 women who had smears taken for cervical cytological examination in Western Australia during an eight-week period in 1983. The peak smear rate was 340.7 per 1000 at 25-29 years of age and fell thereafter with age. The estimated peak frequency of smears that were designated as "screening" smears was 178.3 per 1000 at 30-34 years of age. Screening smears comprised 39% to 66% of the total number of smears, depending on age. After correction for the estimated prevalence of past hysterectomy, only in the age range 20-34 years did the rate of all smears approach the rate of screening smears that would be obtained under a recommended frequency of once every three years. The frequency of screening smears was 20% less in rural areas of Western Australia than in the capital city, Perth. In Perth it fell with decreasing socioeconomic status. General practitioners took 62.4% of all smears and 70.3% of screening smears. On average, female general practitioners took twice as many smears than did male general practitioners.