Isolation and characterization of highly purified mosquito oostatic hormone

Oostatic hormone, the hormone that inhibits vitellogenesis in mosquitoes, was purified 7,000-fold with a recovery of 70% from the ovaries of the mosquito Aedes aegypti. The purification procedure included heat treatment and chromatography on ion exchange and gel filtration columns. The hormone is a small peptidelike molecule of molecular weight 2,200 at pH 4.5, which aggregates into larger molecular species of trimer and octamer at pH 7.0 as determined by gel filtration. The hormone is positively charged at pH 7.8 and has a low Rf at pH 9.4 on disc gel electrophoresis. Injection of purified oostatic hormone (9 ng) into female mosquitoes inhibited yolk deposition and vitellogenin synthesis. Activity of the oostatic hormone in the mosquito ovary increased rapidly following blood feeding and reached a maximum after 48 h. Oostatic hormone of A. aegypti injected into autogenous Aedes taeniorhynchus inhibited egg development. Repeated injections of dilute oostatic hormone at 24 h intervals partially arrested egg development, resulting in 60% reduction in the number of eggs laid. This hormone does not block release of egg development neurosecretory hormone (EDNH) from the mosquito brain but rather appears to act on the ovary.     

A conceptual framework for comparing species assemblages in native and exotic habitats

Exotic (nonnative) species are known to have a wide variety of impacts on native biota. One potential set of impacts that have been poorly studied are the effects of replacing native habitat-providing species with exotic ones, e.g. when native trees that compose a woodland are replaced by an exotic tree plantation. Here we develop a graphical model that can be used to explore how multiple taxonomic components (such as birds, mammals and plants) respond to such changes. We suggest that four categorical responses are possible, with respect to changes in species richness (or other quantitative measures) of taxonomic groups within species assemblages. First, that each taxonomic group compared between habitats will be relatively unchanged, e.g. have equivalent values of species richness. Second, that a decrease (for example in species richness) of one group will be compensated for by an increase (in species richness) of another group. Third, that one or more groups will decrease without any compensated increases in other groups. Fourth, that one or more groups will increase without any compensated decreases in other groups. We provide empirical support for 3 of these 4 responses, with respect to measures of species richness, with much evidence for equivalency between habitats. These types of comparisons should provide a valuable tool for evaluating 1) the efficacy of environmental mitigation efforts that artificially create or restore habitats and 2) the types of changes that have occurred over time or across space as native habitat-producing species are replaced by exotic ones. Finally, this conceptual framework should help to broaden the range of possible changes considered by ecologists who study the impacts of exotic species.     

Biosynthesis and distribution of ecdysone and 20-OH-ecdysone in Aedes aegypti

The distribution and biosynthesis of ecdysone and 20-hydroxyecdysone (20-OH-ecdysone) was followed in sugar- and blood-fed female Aedes aegypti. In both sugar- and early blood-fed animals most of the ecdysteroid determined by radioimmunoassay was found outside the ovary. Twenty-four to 40 h after blood feeding, however, ecdysteroid was distributed between ovary and carcass in the ratio of 1:1.5. Ecdysteroid titer reached a plateau between 18 to 40 h after the blood meal and decreased thereafter. Analysis of the ecdysteroid titer using thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) revealed that both 20-OH-ecdysone and ecdysone were synthesized after the blood meal. The ratio of 20-OH-ecdysone to ecdysone remained essentially constant and fluctuated in parallel throughout egg development. Chromatography of the early ecdysteroid peak (8 h after feeding) using TLC and HPLC indicated that although it cross-reacted with ecdysteroid antibodies, it did not have the same elution times as ecdysone and 20-OH-ecdysone and is, therefore, probably a precursor of these ecdysteroids. Injections of egg development neurosecretory hormone (EDNH) preparation purified to near homogeneity, into ligated abdomens, induced ecdysteroid synthesis only if the abdomens were first treated with methoprene (12.5 pg). Methoprene at this concentration did not stimulate ecdysteroid synthesis in these abdomens. When blood-fed females were treated with [4-¹⁴C] cholesterol and analyzed using TLC and HPLC procedures, both [¹⁴C]labeled ecdysone and [¹⁴C]labeled 20-OH-ecdysone were synthesized in the ratio of 1:1.5. This report is the first to show that both ecdysone and 20-OH-ecdysone are synthesized in vivo in female A. aegypti.     

Analysis of viability and cell types of macroalgal protoplasts using flow cytometry

The ability to rapidly distinguish viable sub-populations of cells within populations of macroalgal protoplast isolations was demonstrated using flow cytometry. Viable protoplasts from Ulva sp. and Porphyra perforata J. Ag. were distinguished from non-viable protoplasts based on differential fluorescein accumulation. The identities of cortical and epidermal protoplasts from Macrocystis pyrifera (L.) C. Ag. were inferred based on light-scattering and chlorophyll a autofluorescence. Three cell types could be distinguished among protoplasts released from thalli of P. perforata based on chlorophyll a and phycoerythrin autofluorescence. Mixed protoplast populations of Ulva sp. and P. perforata were also discernable based on relative chlorophyll a and phycoerythrin autofluorescence. The ability to screen heterogenous protoplast populations rapidly, combined with the cell sorting capabilities of many flow cytometers, should prove valuable for seaweed biotechnology.     

Characteristics of a human epidermal squamous carcinoma cell line at different extracellular calcium concentratrions

A continuous line derived from a human skin squamous cell carcinoma has been grown in media of high, normal and low Ca²⁺ concentrations. The growth rate was unaffected by the Ca²⁺ levels even though morphological changes were observed. Desmosomes were absent at low Ca²⁺ and areas of cell piling were observed at high Ca²⁺. Cell protein staining patterns on polyacrylamide gels were identical for cells grown at the three Ca²⁺ levels. The variations were minor for the glycoproteins reacted with ¹²⁵I-conA. Lactoper-oxidase iodination revealed changes in cell surface proteins, most markedly in the emergence of new proteins at high Ca²⁺.     

Some adaptations between Danaus plexippus and its food plant, with notes on Danaus chrysippus and Euploea core (Insecta: Lepidoptera)

Behaviour of the egg-laying Monarch in captivity suggests that the concentration and quality of cardiac glycosides in the food plant are not important oviposition cues. The presence of eggs (as previously noted by Urquhart, 1960) and larvae feeding on the food plant, act as mild deterrents.
The butterfly's emetic potency (see Table XIII(a)) can sometimes surpass that of the leaves of the host plant itself. Unidentified factors, providing the internal plant environment, are more important as cardiac glycoside storage stimulants than either the quantity or quality of the cardenolides present. In the laboratory D. plexippus oviposited preferentially on a plant with relatively low cardiac glycoside content, but which produced the most powerfully protected (emetic) adult.
Metabolic changes during the pharate pupal stage, but also, in the case of Euploea core , in the larval fifth instar, rather than larval sequestration, may account for the major increase or decrease in butterfly toxicity compared with that of the food plant.
Temperature does not affect the storage of cardenolides except indirectly by altering metabolic rate. There is no evidence to support the concept that current "physiological cost" of cardenolide storage is high. Like the toad, this butterfly can be assumed to have evolved an enzvmatic system well adjusted to the presence of cardenolides in its bodv tissues.