Constitutive Secretion of Acid Hydrolases in Tetrahymena thermophila

THE role of lysosomal enzymes in intracellular digestion is now well established [11]. Most often we think of lysosomal hydrolases in catabolism of endogenous or foreign material taken up by endocytosis. There is however, a number of reports dealing with the release of acid hydrolases into the extracellular fluid in a variety of eukaryote cells. These cells range from Saccharomyces cerevisiae [15], Dictyostelium discoideum [10], Leishmania donovani [20], Acanthamoeba castellani [22], Entamoeba histolytica [12, 31], and species of Tetrahymena [1–3, 6] to mammalian cells in culture [49]. Concerning the latter, fibroblasts and hepatocytes in culture release acid hydrolases to the extracellular medium, but only if the synthesis of a specific recognition marker is impaired in the cells. This marker (man-nose-6-phosphate) is used for receptor mediated segregation of lysosomal enzymes into the lysosomal compartments. If the receptor or the marker are lacking, the hydrolases fail to enter the lysosomal compartment, and are secreted in immature form together with molecules belonging to the constitutive secretory pathway of the cells [8, 49]. Such a release of acid hydrolases seems to occur spontaneously from mammalian osteoclasts [4]. Macrophages, on the other hand, need a specific stimulation for their release process [40]. In lower eukaryotes the release may     

Monophily and pollination mechanisms in Angraecum arachnites Schltr. (Orchidaceae) in a guild of long-tongued hawk-moths (Sphingidae) in Madagascar

Anthecological relations between a long-spurred angraecoid orchid and pollinating Sphingidae are documented for the first time. In a primary forest on the Central Plateau of Madagascar Angraecum arachnites Schltr. was found to be pollinated by and adapted to a single species of hawk-moth, Panogena lingens (Butler), despite abundance of many concurrent Sphingidae of which several were also long-tongued. Furthermore, P. lingens was dimorphic in the length and breadth of its proboscis and only the morph with the longest and most slender proboscis was recorded to pollinate A. arachnites . Exclusive and precise adaptation to the latter morph of P. lingens existed in floral morphology and probably in other characteristics such as flowering phenology and chemical signalling. Several concurrently flowering orchid species were sharing P. lingens as a pollinator resource. The monophily in A. arachnites is interpreted as a result of a refined long-term specialization developed within an archaic evolutionary relationship in a relatively stable environment. The extraordinary number and diversity of long-spurred Orchidaceae in Madagascar appears to be a direct coevolutionary consequence of an Old-World-unique diversity of long-tongued archaic Sphingidae that has persisted in this isolated land.     

Ectomycorrhizal fungal communities in late-successional Swedish boreal forests, and their composition following wildfire

This study was conducted to evaluate the effects of wildfires on ectomycorrhizal (EM) fungal communities in Scots pine ( Pinus sylvestris ) stands. Below- and above-ground communities were analysed in terms of species richness and evenness by examining mycorrhizas and sporocarps in a chronosequence of burned stands in comparison with adjacent unburned late-successional stands. The internal transcribed spacer (ITS)-region (rDNA) of mycobionts from single mycorrhizas was digested with three restriction enzymes and compared with an ITS–restriction fragment length polymorphism (RFLP) reference database of EM sporocarps. Spatial variation seemed to be more prominent than the effects of fire on the EM fungal species composition. Most of the common species tended to be found in all sites, suggesting that EM fungal communities show a high degree of continuity following low-intensity wildfires. Species richness was not affected by fire, whereas the evenness of species distributions of mycorrhizas was lower in the burned stands. The diversity of EM fungi was relatively high considering that there were only three EM tree species present in the stands. In total, 135 EM taxa were identified on the basis of their RFLP patterns; 66 species were recorded as sporocarps, but only 11 of these were also recorded as mycorrhizas. The species composition of the below-ground community of EM fungi did not reflect that of the sporocarps produced. EM fungal species present in our ITS–RFLP reference database accounted for 54–99% of the total sporocarp production in the stands, but only 0–32% of the mycorrhizal abundance.     

Effects of Azide on Photophosphorylation in Isolated Spinach Chloroplasts

The influence of sodium azide on open-chain and flavine mononucleotide mediated cyclic photophosphorylation in isolated spinach chloroplasts was investigated under anaerobic conditions. Open chain phosphorylation was completely inhibited with DCMU both in the presence and absence of sodium azide in the experimental medium. Flavine mononucleotide mediated photophosphorylation was only slightly inhibited by DCMU in the absence of sodium azide but inhibited in two steps by increasing amounts of DCMU when sodium azide was present in the medium. The first step can be explained as being mainly an effect of DCMU on an open chain electron transport, with water and H₂O₂ as electron donors and with flavine mononucleotide — kept in an oxidized state by sodium azide — as the electron acceptor. The second step, as well as the comparatively insensitivity to DCMU in the absence of sodium azide, depends on cyclic photophosphorylation mediated by flavine mononucleotide.     

THE INTRINSIC MECHANISM OF CHROMATIN DECONDENSATION AND ITS ACTIVATION IN HUMAN SPERMATOZOA*

In vitro decondensation of human sperm chromatin induced by the activation of an intrinsic mechanism was studied by light microscopy, scanning and transmission electron microscopy. Morphological evidence was provided to support the concept that this mechanism is essential for the chromatin decondensation occurring in vivo. Prostatic zinc is hypothesized to preserve this potential decondensation ability from oxidative destruction, by reversibly binding to free thiol-groups. The unique occurrence of disulphide-stabilized structures in eutherian spermatozoa may serve to protect the spermatozoon from structural degradation by its own proteolytic activity during the relatively slow passage through the eutherian egg investments.     

Does Chloroquine,an Antimalarial Drug,Affect Autophagy in Tetrahymena pyriformis?

ABSTRACT. The effect of chloroquine (CQ) on autophagy was studied in starved Tetrahymena pyriformis . When a proliferating Tetrahymena culture is transferred to a starvation medium, autophagy commences although cells most advanced in the cell cycle will divide. The drug was added to 1-h starved cells at different pH values because CQ affects pH dependently. The CQ concentration blocking all cell divisions was determined as the lowest toxic, but sublethal, concentration. Hence, the highest tolerated concentrations at pH 6.8, 7.1, and 7.7 were 1.0, 0.3, and 0.03 mM CQ, respectively. Lower CQ concentrations had a dose-dependent effect on cell increment and higher concentrations induced cell mortality. Rates of cell motility and decreases in cell volume were affected by the drug, while the capacity for endocytosis was unaffected in low concentrations but affected dose dependently in high concentrations. Light microscopically, all drug-treated cells contained small refractive bodies, but in toxic concentrations they also contained conspicuously large vacuoles. After 1 h and 4 h in CQ, fine structure analysis showed autophagosomes with electron-dense material in cells in tolerated concentrations and of enlarged size, but decreased number, in toxic concentrations. The contents of autophagosomes revealed cell organelles in different stages of disintegration. The conclusion is that the drug enhances autophagy in Tetrahymena in a pH-, dose-, and time-dependent manner.