The cytonucleus test in the rat: a combined metaphase and micronucleus assay

The suitability of the rat as a species choice for the micronucleus assay and the possibility of combining both metaphase and micronucleus analysis using one set of animals were investigated. Cyclophosphamide, trenimon, vinblastine sulphate and dimethyl benzanthracene were used to optimise the study design and experimental procedures. The sample times ranged from 12 to 72 h after a single dose of the compound. A maximal response in both micronucleus and chromosome aberration induction was observed 24 h after dosing. This sample time is recommended as the single sample time for the screening of all classes of compound. Using the optimal conditions, a number of different mutagens/carcinogens were then analysed for the induction of both micronuclei and chromosome aberrations in the same animal. The compounds selected were mitomycin C, methotrexate, 5-fluorouracil, hexamethyl phosphoramide, benzo[a]pyrene, benzidine and diaminoterphenyl. The results show that the rat is a responsive test species and that it is possible to combine both metaphase and micronucleus analysis in the same animal.     

Formation and regeneration of Frankia protoplasts

A technique for forming protoplasts from Frankia cells and regenerating them to the normal hyphal mode of growth is described. Electron microscopy proved that protoplasts were studied and not spores or small hyphae. Regenerated colonies were investigated for genetic markers. One ArI3 colony had been cured of its plasmids without being affected in its symbiotic properties.     

A conformational rearrangement in gramicidin A: from a double-stranded left-handed to a single-stranded right-handed helix.

A conformational transition is described for the polypeptide, gramicidin A, in which a dimer that forms a left-handed intertwined antiparallel helix is converted to a single-stranded amino terminus to amino terminus right-handed helix. The starting structure is determined here by solution NMR methods while reference is made to the well-established folding motif of gramicidin in a lipid bilayer for the ultimate conformation of this transition. Furthermore, an organic solvent system of benzene and ethanol in which gramicidin has a unique conformation is identified. This conformation is shown to be very similar to that derived from X-ray diffraction of crystals prepared from a similar solvent system.     

Feeder cells enhance oncolytic and proliferative activity of long-term human bone marrow interleukin-2 cultures and induce different lymphocyte subsets

Summary The effect of feeder cells on oncolytic activity of lymphocyte subsets and their growth was evaluated in long-term human bone marrow interleukin-2 (IL-2) cultures. Two B-lymphoblastoid cell lines (Daudi and Epstein-Barr-virus-transformed BSM) and two human leukemias, AML-M5, were used as feeder cells. The most prominent effects were seen in cultures stimulated with Daudi cells. In these cultures, cytotoxic activity was 100–1000 times increased against a broad range of target cells and the total cellular expansion was more than 40 times higher than in control cultures. This Daudi-related effect appeared to be mediated by natural killer (NK) cells, since cellular expansion occurred mostly in the CD16⁺ and CD56⁺ CD3^– NK cell subset. In cultures stimulated with BSM and acute myelogenous leukemia (AML) feeder cells, the increase in proliferation was similar, but the enhancement of cytotoxicity, even though significant, was less prominent. Although all feeder cells were effective in stimulation of bone marrow reactivity, the highest cytotoxicity was always observed with feeder cells autologous to the targets, indicating some degree of specificity. This was especially evident in cultures stimulated with autologous versus allogeneic AML feeder cells. In contrast to Daudistimulated IL-2 cultures, in which the highest expansion of CD3^– CD56⁺ NK cells was observed, in BSM and AML cultures, the CD3⁺ CD56^+/- T cell subsets were more prolific. This indicates that the response and phenotypic heterogeneity of bone marrow cultures depends on the type of feeder cells used. This observation indicates that the preferential stimulation of a pertinent lymphocyte subset for therapeutic purposes may be possible.Recipient of Florence Maude Thomas Cancer Research Professorship     

Regulation of the trimethylamine N-oxide (TMAO) reductase in Escherichia coli: analysis of tor::Mud1 operon fusion

Summary Mud1 insertion mutants of Escherichia coli were obtained in which the lac structural genes were fused to the promoter of torA, a gene encoding the trimethylamine N-oxide (TMAO) reductase. Expression of the fusion is induced by TMAO and repressed by oxygen. However, in contrast to the nar operon which codes for the nitrate reductase structural genes, the tor::Mud1 fusion was found to be independent of the positive control exerted by the nirR gene product and not repressed by the molybdenum cofactor. The torA gene which is strongly linked to pyrF (28.3 U) is different from any tor gene already described in E. coli or in Salmonella typhimurium.     

THE CLASSIFICATION OF PROBOSCIDEA: HOW MANY CLADISTIC CLASSIFICATIONS?

Abstract— Hennig conceived a method to build a "phylogenetic system", with the stipulation that a "properly drawn phylogenetic tree must be directly translatable into the language of phylogenetic systematics". Consequently, this system could be the general reference system of biology. A review of the classificatory technical improvements, conventions and rules which have been proposed for the past twenty years together with their application to the classification of the Proboscidea, leads to the conclusion that more than one formal system can be built upon one given cladogram. As words are used more frequently for communication than diagrams, schemes or graphs, the "general reference system of biology1' remains somewhere in Utopia. The "phylogenetic system" is rather more synonymous with a cladogram than with a written classification.     

Optimization of the bare platinum electrode as an oxygen measurement system in photosynthesis

This paper is concerned with the definition of the standard conditions required for optimum operation of the bare platinum electrode with photosynthetic samples. Experimental evidence shows the following: 1) Polarization circuits should have zero resistance; 2) The electrolyte layer between the electrodes should have a conductance higher than 54×10^{–6 –1} per mm² of platinum electrode area; 3) The electrodes should be polarized just before taking the measurements. All these facts can be interpreted in terms of phenomena occurring on the electrode: The adsorption of hydrogen on the electrode imposes the need for low resistances in the system, and oxygen consumption by the electrode is minimized by polarizing the electrodes as late as possible. This investigation increases the reliability of the bare platinum electrode and gives a basis for a comparison of the results from different experiments. Demonstrations of the pertinence of these conditions are made in our lab with the algae Dunaliella Tertiolecta.     

Coculture of human keratinocytes and melanocytes: differentiated melanocytes are physiologically organized in the basal layer of the cultured epithelium

Human epidermal keratinocytes differentiate in vitro into a stratified epithelium suitable for grafting on burned patients. In this paper, we show that differentiated melanocytes are present in the cultured epithelium. In particular, we have found that i) melanocytes proliferate in the same culture conditions that allow keratinocyte growth, ii) during the culture the ratio between keratinocytes and melanocytes tends to remain constant, iii) melanocytes organize into the basal layer of the cultured epithelium independently of the presence of dermis, develop dendritic arborizations with melanosome-containing processes and transfer melanosomes into keratinocyte cytoplasm.     

Evidence for a linear variation of the miss and single hit S-state probabilities with the flash number,measured by oxygen evolution in Dunaliella tertiolecta

Oxygen evolution in Dunaliella tertiolecta under flashing light was measured with a bare electrode, at a 10 Hz frquency. The sigma coefficients of the oxygen evolution recurrence law (Thibault (1978) J Theor Biol 73, 271) were determined using groups of nine consecutive points. The S-state transition probabilities were computed from the sigma coefficients and plotted as a function of the flash number of the first of the points used. Low standard deviations over the sigma coefficients resulted from the use of our system (Meunier & Popovic (1988) Rev Sci Instr 59, 486). We observed a linear lowering of the miss probability with time, and a linear rise of the single-hit probability with the same absolute value of the slope. The hypothesis that the slopes were zero was statistically tested and was rejected with a 99.9% confidence interval. Our work demonstrates that, to be accurate, an oxygen evolution model has to take the variations in the properties of S-states into account.International Journal of Fracture 76 (1995) R37