Polyamine Depletion Following Exposure to DL-α-Difluoromethylornithine Both In Vivo and In Vitro Initiates Morphological Alterations and Mitochondrial Activation in a Monomorphic Strain of Trypanosoma brucei brucei

ABSTRACT. DL-α-difluoromethylornithine (DFMO), a specific irreversible inhibitor of ornithine decarboxylase (ODC), rapidly depletes cells of intracellular putrescine. When administered to animals and humans, DFMO cures acute infections of trypanosomiasis. In order to determine if the mechanism of drug action is related to initiation of transformation and biochemical alterations subsequent to polyamine depletion, trypanosome morphology and mitochondrial activation were studied in a monomorphic strain of Trypanosoma brucei brucei. Exposure of trypanosomes to DFMO in vivo in infected rodents or in vitro in culture resulted in a depletion of intracellular putrescine and a cessation of cell division without specific cytotoxicity. These events were followed by a transformation of the long slender bloodstream form to a short stumpy form via an intermediate morphology. Putrescine, the product of the ODC reaction, abrogates this effect. When introduced into SDM-79 medium, the intermediate form is capable of further transformation to an "insect" procyclic trypomastigote whereas the long slender form and short stumpy form are not. Short stumpy forms are incapable of binary fission and have lost their infectivity for the vertebrate host. In addition, the mitochondrial marker enzyme, NAD diaphorase, was found only in the short stumpy and intermediate forms. We hypothesize that the short stumpy phenotype may not be a viable stage in the natural transformation of the trypanosome from its mammalian host to the insect vector.     

Separation and Purification of Protoplast Types from Commelina communis L. Leaf Epidermis

Guard cell and epidermal/subsidiary cell protoplasts obtainedby enzymic digestion of peeled Commelina communis leaf epidermiswere separated and purified by discontinuous density gradientccntrifugation with media based on Percoll (Pharmacia Fine ChemicalsAB, Uppsala, Sweden). The cell types were recovered over 99.9%pure at yields exceeding 50% efficiency, and mesophyll contaminationcould be virtually eliminated when desired. Osmotic characteristicsof the protoplast types were evaluated and compared to in vivovalues, and the viability of the protoplasts, assessed usinga range of criteria, was found to be high. Purified Commelinaguard cell protoplasts were able to evolve O₂ when illuminated,and this was substantially reduced in the presence of the inhibitorDCMU, indicating that they possess photosystem II activity.Specific advantages of this method of protoplast purification,and the potential uses of separate suspensions of guard cellsand epidermal/subsidiary cells in experiments on stomatal physiologyare discussed. Key words: Commelina communis, Protoplasts, Epidermis     

Effect of Diurnally Fluctuating vs. Constant Temperatures on Flower Induction and Sex Expression in the Olive (Olea europaea)

Olive trees must be exposed to a period of winter chilling temperatures in order to form inflorescences the following spring. The effects of diurnally fluctuating and constant temperatures on flower induction and sex expression in the olive were compared under controlled environment conditions. The effect on flowering of diurnally fluctuating temperatures depends upon the length of time at the higher temperatures. While daily exposure to the high temperatures (21° to 26°C) for a short period (four hours) intensified the effectiveness of the low temperatures (7° to 12.5°C), when the high temperature duration was 12 hours it counteracted the low temperature action. Possibly, daily low temperatures act to stimulate reactions leading to floral hormone synthesis, while daily short exposures to high temperatures act to maintain metabolic activity, promote energy-yielding reactions, and possibly stimulate cell-division activity. In the olive, an intermediate constant temperature (12.5°C) favors both types of reactions and induces flowering; however, in this case, the pistils fail to develop. The olive has very specific temperature requirements for flowering and neither the mean temperature nor the accumulated number of hours below a given value, e.g. 7°C, adequately characterizes these requirements.     

Additional Nucleotide Sequences in Precursor 16S Ribosomal RNA from <Emphasis Type="Italic">Escherichia coli</Emphasis>

MATURE 5S, 16S and 23S ribosomal RNA species present in E. coli ribosomes are the end products of complex biosyn-thetic pathways. They are formed by reduction in length, and methylation of longer RNA chains transcribed on the ribosomal RNA cistrons of E. coli DNA. While these modifications take place the ribosome structure is formed by progressive addition of ribosomal proteins and conformational changes in the resulting ribonucleoprotein precursor particles¹.     

Ultrastructural Observations on Life Cycle Stages of Pneumocystis carinii

An ultrastructural study of life cycle stages of Pneumocystis carinii in infected rat lungs in situ was undertaken utilizing 8 different modes of fixation. Three of the fixatives employed gave good fixation of cysts and intracystic bodies, but for the trophic forms fixation was only fair. Both the trophic forms and intracystic bodies have nuclear pores. The mitochondria of the organism have cristac that appear lamellar. One of the fixation modes revealed a thin, electron-dense layer on the outer surface of the cell wall, a "fuzzy coat" that had not been described previously. This material appears to mediate tight adhesion of trophic forms with other trophic forms, cysts, and with pneumocytcs of the lung alveolus.     

Hyolith guts in the Cambrian of northern Australia -turning hyolithomorphs upside down

A single specimen of the early Middle Cambrian (Ordian) orthothecimorph hyolith Gudugu-wan hardmani with preserved gut filling has been discovered in the Montejinni Limestone, Wiso Basin, northern Australia. This represents only the eighth such occurrence worldwide. The specimen is unique among orthothecimorphs in retaining the gut filling in combination with the external mould of the operculum and invites revision of the accepted soft-part recon struction in this group, whereby the opercular lateral processes are now recognized to be ventral. Thus orthothecimorph conchs, if horizontal in life, would have been oriented ventral side down, while hyolithomorphs adopted the reverse orientation: horizontal, ventral side up.     

Light-limited growth and photosynthetic characteristics of two planktonic desmid species

1. Staurastrum chaetoceras, originating from a eutrophic environment, and Cosmarium abbreviatum var. planctonicum, from an oligo-mesotrophic one, were grown at 20°C in continuous-flow culture under varied light conditions. 2. Maximum growth rate in S. chaetoceras was approximately 50% higher than in C. abbreviatum, corresponding to a higher photosynthetic capacity. 3. Higher dry weight values and dark respiration rates in C. abbreviatum (compared with S. chaetoceras) support the suggestion from growth-irradiance curves that C. abbreviatum has to invest a greater part of its photosynthetic energy in the construction and maintenance of specific cell components. 4. The differences in growth and photosynthetic parameters might be related to the presence of a copious mucilaginous cell envelope in C. abbreviatum; such an envelope is lacking in S. chaetoceras. 5. The possible ecological significance of these species-specific differences is discussed.