Structural and Functional Impact of SRP54 Mutations Causing Severe Congenital Neutropenia

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Conditionality of phase resetting by inhibitors of 80 S translation inGonyaulax polyedra

Summary Gonyaulax polyedra was subjected to a cold treatment of 18 h at 10 °C leading to arrhythmicity. Subsequently, the circadian rhythm of bioluminescence was investigated at the permissive temperature of 20 °C. 1-h pulses of 10 M cycloheximide or 2 M anisomycin, when given after the temperature step-up, resulted only in a very weak resetting of the circadian oscillator, in marked contrast to the behaviour of cells kept continuously in oscillatory conditions at 20 °C. The extremely reduced sensitivity to 80 S inhibition was characteristic for the first cycle after the temperature step-up, whereas cells treated with cycloheximide in the second cycle after re-initiation of rhythmicity showed a gradual recovery of resettability, though the phase response curve was still atypical; treatment in the 3rd cycle after step-up led to a relatively normal phase response curve. The observed insensitivity in the 1st cycle was neither a consequence of insufficient drug action, nor of a transient non-oscillatory behaviour after temperature step-up. Already in the first hours after transfer to 20 °C, 80 S translation was strongly suppressed by cycloheximide, and the cells were also efficiently reset by changes of the light-dark zeitgeber. Resettability of the circadian oscillator by 80 S inhibitors is, therefore, conditional.Abbreviations Ani anisomycin - Chx cycloheximide - CT circadian time - LD light-dark cycle - LL constant light - TCA trichloroacetic acid This work was supported by the Deutsche Forschungsgemeinschaft     

Coupling between phosphoinositide breakdown and early mitogenic events in fibroblasts. Studies with fluoroaluminate, vanadate, and pertussis toxin

In the preceding paper (Paris, S., and Pouysségur J. (1987) J. Biol. Chem. 262, 1970-1976), AlF4- and vanadate have been shown to induce inositol phosphate formation in resting hamster fibroblasts (CCL39). In this study, we show that these two phosphate analogs are good tools to explore the causal relationship between phosphoinositide breakdown and early mitogenic events. AlF4- can activate, very similarly to the mitogen alpha-thrombin: the amiloride-sensitive Na+/H+ antiport, the bumetanide-sensitive Na+/K+/Cl- co-transport, and the expression of c-myc mRNA. The link between phospholipase C activation and these early events of the mitogenic response is demonstrated by the similarity of all dose-response curves for NaF and AlCl3 and by the common sensitivity of the four events to pertussis toxin. Vanadate likewise stimulates the Na+/H+ antiport through a pertussis toxin-sensitive pathway. On longer incubations, both fluoride and vanadate were found to be toxic and failed to induce DNA synthesis. Therefore, we have used pertussis toxin to investigate the link between phospholipase C activation and commitment to DNA synthesis. We show that pertussis toxin strikingly inhibits thrombin-induced reinitiation of DNA synthesis but does not affect the stimulation by the epidermal or fibroblast growth factors, two mitogens that do not stimulate phosphoinositide breakdown in CCL39 cells. In conclusion, these studies demonstrate that activation of phospholipase C, if not an obligatory step in the action of all growth factors, plays a crucial role in the mitogenic signaling pathway of alpha-thrombin.     

Nasal allergy to fungi in guinea pigs

Sensitized guinea pigs produced specific IgG and IgE antibodies toward Cladosporium and Alternaria. In presence of fungal extracts, nasal mast cells degranulate. Ultrastructural modifications of the cells during degranulation have been established. The ciliary epithelium and the ciliary beating are not affected by fungal allergens.     

Kinetic Concepts for Measuring Microbial Rate Constants: Effects of Nutrients on Rate Constants

We investigated the effect of preincubation of environmental waters amended with inorganic nutrients (nitrogen, phosphorus, and traces of iron and magnesium) on the kinetics of the microbial transformation of phenol, propanil, propyl ester of (2,4-dichlorophenoxy)acetic acid, methyl parathion, Ronnel, and methoxychlor in pond and river waters. No effect on the second-order rate constants for these compounds was observed, although there was an increase in the bacterial populations and the pseudo-first-order rate constants. The use of nutrient-amended waters could be a useful tool for estimating second-order rate constants for an expanded number of compounds. This technique would provide a larger data base for predicting the behavior of xenobiotic compounds in the environment by using currently available mathematical models.