De novo-synthesized ceramide signals apoptosis in astrocytes via extracellular signal-regulated kinase.

Recent observations support the importance of ceramide synthesis de novo in the induction of apoptosis. However, the downstream targets of de novo-synthesized ceramide are unknown. Here we show that palmitate incorporated into ceramide and induced apoptotic DNA fragmentation in astrocytes. These effects of palmitate were exacerbated when fatty acid breakdown was uncoupled and were not evident in neurons, which show a very low capacity to take up and metabolize palmitate. Palmitate-induced apoptosis of astrocytes was prevented by L-cycloserine and fumonisin B1, two inhibitors of ceramide synthesis de novo, and by PD098059, an inhibitor of the extracellular signal-regulated kinase (ERK) cascade. Accordingly, palmitate activated ERK by a process that was dependent on ceramide synthesis de novo and Raf-1, but independent of kinase suppressor of Ras. Other potential targets of ceramide in the control of cell fate, namely, c-Jun amino-terminal kinase, p38 mitogen-activated protein kinase, and protein kinase B, were not significantly affected in astrocytes exposed to palmitate. Results show that the Raf-1/ERK cascade is the selective downstream target of de novo-synthesized ceramide in the induction of apoptosis in astrocytes and also highlight the importance of ceramide synthesis de novo in apoptosis of astrocytes, which might have pathophysiological relevance.     

Morphological characteristics in relation to diet in five coexisting Thai fish species

Morphological characteristics and intestinal content were analysed for five species of coexisting freshwater fishes in Thailand: Rasbora caudimaculata , Schistura desmotes , Dermogenys pusillus , Xenentodon cancila and Monopterus albus (all found in riffle habitats in Thai streams). Rasbora caudimaculata , S. desmotes and D. pusillus fed predominantly on ephemeropterans, hymenopterans and dipterans, X. cancila fed predominantly on fishes, and larger aquatic invertebrates such as Odonata, and M. albus fed on detritus as well as invertebrate prey such as crustaceans and Odonata. Intestine length, mouth height, mouth width, eye position and mouth orientation varied among all five species. Canonical analysis of discriminance of mouth height, width and intestine length showed a clear dispersion of species, which was supported by intestine content. Evolutionary processes leading to the present differences in morphological characters resulted in each of the five species consuming a different portion of the available resource base, thereby facilitating coexistence.     

Effect of thiostrepton and 3′-terminal fragments of aminoacyl-tRNA on EF-Tu and ribosome-dependent GTP hydrolysis

The effect of the antibiotics thiostrepton and micrococcin on EF-T_u-catalyzed (ribosome-dependent) GTP hydrolysis in the presence of A-Phe, C-A-Phe, or C-C-A-Phe (related to the sequence of the 3′-terminus of aminoacyl-tRNA)(System I) or by methanol (‘uncoupled GTPase’, System II) was investigated. In System I, thiostrepton increases the binding affinities of the effectors to the EF-T_u·GTP·70 S ribosome complex, as well as the extent of the GTP hydrolysis, while the K^GTP_m is virtually unchanged. Similarly, in the uncoupled system (System II) and in the absence of effectors, thiostrepton significantly increases V^GTP_max, whereas K^GTP_m remains unaffected. Micrococcin is without any effect in both systems. The ‘uncoupled GTPase’ (in System II) is also strongly inhibited by C-A-Phe. The results indicate the crucial role of the EF-T_u site which binds the aminoacylated C-C-A terminus of aminoacyl-tRNA in promoting GTP hydrolysis. It follows that the binding of the model effectors (such as C-C-A-Phe) to that site is favorably influenced by the interaction of thiostrepton with the 50 S ribosomal subunit, whereas thiostrepton, per se, does not influence the affinity of EF-T_u for GTP.     

Genotoxicity of ptaquiloside, a bracken carcinogen, in the hepatocyte primary culture/DNA-repair test

The genotoxicity of ptaquiloside (PT), recently isolated from bracken fern and shown to be carcinogenic, was examined by means of the hepatocyte primary culture/DNA-repair test. PT elicited clear unscheduled DNA synthesis with a dose-response effect. The result indicates that PT is a genotoxic carcinogen.     

Establishment and Characterization of Cell Lines from Homozygous Brachyury (T/T) Embryos of the Mouse

Lethal mutations in the T/t complex cause stage-specific morphologic abnormalities during early embryogenesis of mice. Although mutant embryos are lethal at the early stages of development, we have succeeded in establishing several cell lines from one of these mutants ( T/T ). Mutant-specific abnormality was not observed in gross morphology and growth patterns of cells. They, however, retained the characters of freshly dissociated embryonic cells to form smaller aggregates than the wild-type in rotation-mediated aggregation.
One of the T/T cell lines (T-1) formed tumors when injected into one-day-old syngeneic and allogeneic host, Expression of H-2 antigens was serologically studied with H-2 specificity 5 as a marker antigen. All lines except T-1 were shown to have this specificity.