Alpha-terthienyl is toxic to late embryonic stages ofRana pipiens in the presence of sunlight. Neither α-terthienyl alone in the dark nor a previously photolyzed solution of α-terthienyl has comparable activity. The LC50 was 0.11 ppm with 30 min of exposure and 0.018 ppm with 2 hr of exposure to sunlight. Anthracene, a representative example of polycyclic aromatic hydrocarbons widely distributed in the environment, also showed similar phototoxicity, with an LC50 of 0.065 ppm after 30 min of exposure and 0.025 ppm after 5 hr.
SYNOPSIS. Microstome →macrostome transformation in Tetrahymena vorax was induced by suspending microstomes in a transforming principle, stomatin, released by a potential prey, T. pyriformis. It was found that 70–90% of the microstomes formed macrostomes within 7 hours following suspension in this transforming principle. Macrostome formation occurred by the process of oral replacement. This process involved resorption of the microstome oral apparatus and its replacement with a larger (macrostome) one, which arose from an anarchic field that formed behind the resorbing oral area. Ninety‐five percent of those microstomes which were destined to form macrostomes were in some stage of oral replacement 195 minutes after their suspension in stomatin. Several commercially produced products were tested over a wide range of concentrations to determine their ability to act as an inducer of macrostomes. Only 2, Trypticase and Bactocasitone, had any activity, and it was too small to be considered really effective. An attempt was also made to destroy the activity of stomatin by using enzymes. RNAse was effective but only in very high concentrations, so it was suggested that this activity might be related to the destruction of RNA within the transforming cell and not related to hydrolysis of stomatin. None of the other enzymes tested had any effect in reducing the activity of stomatin.
The stalked, ciliated protozoan Vorticella convallaria possesses a highly contractile cytoskeleton consisting of spasmonemes and myonemes. The major component of these contractile organelles is the calcium-binding protein(s) called spasmin. Cloning and characterization of spasmin would help elucidate this contractile system. Therefore, enriched spasmoneme protein preparations from these contractile stalks were used to produce a monoclonal antibody to spasmin. A monoclonal antibody, 1F5, was obtained that immunolocalized specifically to the spasmonemes and the myonemes and recognized a 20-kD calcium-binding protein in spasmoneme protein preparations. A putative spasmin cDNA was obtained from a V. convallaria cDNA library and the derived amino acid sequence of this cDNA revealed an acidic, 20-kD protein with calcium-binding helix-loop-helix domains. The physical properties of the putative spasmin were assessed by characterization of a recombinantly-produced spasmin protein. The recombinant spasmin protein was shown to bind calcium using calcium gel-shift assays and was recognized by the anti-spasmin antibody. Therefore, a V. convallaria spasmin was cloned and shown to be a member of the EF-hand superfamily of calcium-binding proteins.
The polymorphic ciliated protozoan Tetrahymena vorax can undergo differentiation from the microstomal form, which normally feeds on bacteria and other particulate matter, into the macrostomal cell type, which is capable of ingesting prey ciliates. The process is triggered by exposure of the microstome to an inducer contained in stomatin, an exudate of the prey. To establish the identity of the signal, stomatin was fractionated by combinations of cation exchange, HPLC, and TLC, and the fractions were assayed for biological activity. Although no single active fraction of purified inducer was obtained, all fractions with activity contained ferrous iron and the nucleic acid catabolites hypoxanthine (6-oxypurine) and uracil (2,4-dioxopyrimidine), probably in a chelated form. The activity of synthetic complexes containing these three components is equivalent to stomatin. These results indicate a role for ferrous iron and its potential in chelated form to signal differentiation in certain protozoa and, perhaps, in other organisms as well.
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