The BULT Melanoma: A Spontaneous Transplantable Tumor in Mice

The BULT melanoma originated at Brown University as a spontaneous, small black nodule on the tail of an adult female mouse of the LT/Ch strain. Histological examination of a portion of the tumor indicated that it was intradermal and consisted predominantly of heavily melanized, ovoid to fusiform cells with melanin-laden macrophages scattered among them. The BULT melanoma has been maintained in LT/Ch mice for approximately 5 years by periodic transplantation, at first subcutaneously on the flanks and, more recently, intramuscularly in the hind legs. The shift in transplantation site was made following a marked decline in the growth of subcutaneous grafts. The transplants have retained the uniform deep-black melanization and general histology of the primary melanoma. Numerous melanosomes at all stages of development are found within the melanoma cells. DOPA-positive cytoplasmic vesicles are abundant. Occasional autophagic vacuoles containing clusters of melanosomes are also present. A few metastases from the transplanted melanoma have been observed in lymph nodes and on one occasion in the lungs. When grown in vitro, BULT melanoma cells do not require special growth promoting agents (e.g., TPA; cAMP) in order to proliferate. The BULT melanoma differs in one or more respects from each of the other three transplantable spontaneous mouse melanomas widely used in cancer research. In addition, it arose in a strain of mice characterized by the spontaneous death of melanocytes while the latter are engaged in synthesizing eumelanin within hair follicles. Karyotypic analysis of cultured cells showed a modal chromosome number of 68 with a range of 58–72 chromosomes.     

Transient over-expression of barley BAX Inhibitor-1 weakens oxidative defence and MLA12-mediated resistance to Blumeria graminis f.sp. hordei

BAX Inhibitor-1 (BI-1) is a conserved cell death suppressor protein. In barley, BI-1 ( HvBI-1 ) expression is induced upon powdery mildew infection and when over-expressed in epidermal cells of barley, HvBI-1 induces susceptibility to the biotrophic fungal pathogen Blumeria graminis . We co-expressed mammalian pro-apoptotic BAX together with HvBI-1, and the mammalian BAX antagonist BCL-X_L in barley epidermal cells. BAX expression led to cessation of cytoplasmic streaming and collapse of the cytoplasm while co-expression of HvBI-1 and BCL-X_L partially or completely, respectively, rescued cells from BAX lethality. When B. graminis was attacking epidermal cells, a green fluorescent protein fusion of HvBI-1 accumulated at the site of attempted penetration and was also present around haustoria. Over-expression of HvBI-1 in epidermal cells weakened a cell-wall-associated local hydrogen peroxide burst in a resistant mlo -mutant genotype and supported haustoria accommodation in race-specifically resistant MLA12 -barley. HvBI-1 is a cell death regulator protein of barley with the potential to suppress host defence reactions.     

STUDIES ON THE SYNERGISTIC EFFECT OF PIPERONYL BUTOXIDE AND ISOBUTYLUNDECYLENEAMIDE ON PYRETHRINS AND ALLETHRIN

Experiments are described to compare the toxicity of natural pyrethrins and allethrin, the completely synthetic homologue of cinerin I, and to show the effect of the two synergists, piperonyl butoxide and iaobutyl undecyleneamide (IN 930) on both these active ingredients, using a measured-drop technique with Mucca domestiur L. the housefly, and a residual-film technique with Cimex lectularius L. the bed bug. In the conditions of the experiment, pyrethrins were shown to be twice as toxic as allethrin to flies, and 5.5 times as toxic as allethrin to bugs. The two synergists were tested at several ratios to the two insecticides, ranging from 1 : 1 to 20 : 1. The results were plotted as series of log. concentration/probit regression linea. These were parallel for the bug tests; but in the fly tests, the slope of the line increased with a rise in the proportion of Synergist to insecticide. The estimated median lethal concentrations indicated, in all cases, that the toxicity increased with a rise in the ratio of synergist to insecticide, at least up to 20 : 1. However, the enhancement of toxicity was greatest for the smaller ratios and fell off as the ratio increased. Piperonyl butoxide was the more powerful synergist, increasing the potency of pyrethrins 5 times and allethrin 4 times to flies, and pyrethrins twice and allethrin 3 times to bugs, whereas IN 930 did not increase the potency of either ingredient more than twice to either test insect.
The addition of piperonyl butoxide to residual films of pyrethrins greatly prolonged their effectiveness; but an experiment designed to investigate the effect of the synergist on the stability of this insecticide showed that this action, if it exists, must be slight.     

Soluble nitrogen and phosphorus excretion of exotic freshwater mussels (Dreissena spp.): potential impacts for nutrient remineralisation in western Lake Erie

1. Recent increases in phytoplankton biomass and the recurrence of cyanobacterial blooms in western Lake Erie, concomitant with a shift from a community dominated by zebra mussels (Dreissena polymorpha) to one dominated by quagga mussels (D. bugensis), led us to test for differences in ammonia‐nitrogen and phosphate‐phosphorus excretion rates of these two species of invasive molluscs. 2. We found significant differences in excretion rate both between size classes within a taxon and between taxa, with zebra mussels generally having greater nutrient excretion rates than quagga mussels. Combining measured excretion rates with measurements of mussel soft‐tissue dry weight and shell length, we developed nutrient excretion equations allowing estimation of nutrient excretion by dreissenids. 3. Comparing dreissenid ammonia and phosphate excretion with that of the crustacean zooplankton, we demonstrated that the mussels add to nitrogen and phosphorus remineralisation, shortening nitrogen and phosphorus turnover times, and, importantly, modify the nitrogen and phosphorus cycles in Lake Erie. The increased nutrient flux from dreissenids may facilitate phytoplankton growth and cyanobacterial blooms in well‐mixed and/or shallow areas of western Lake Erie.     

The Physiological Mechanisms of Acclimatization in Tropical Reef Corals

SYNOPSIS. The ability of scleractinian corals to survive changesthat are predicted in the global environment over the next centurywill lie in their physiological mechanisms of acclimatization.Corals display rapid modifications in behavior, morphology andphysiology enabling them to photoacclimate to changing lightconditions, a scenario that demonstrates considerable biologicalflexibility. Here we argue that the acclimatization mechanismsin corals are fundamentally similar to those exhibited by otherinvertebrate taxa. We discuss protein metabolism as a mechanismunderlying acclimatization responses in reef corals, and explorethe relationship between protein turnover, metabolic rate, growthrate, and acclimatization capacity. Our preliminary analysessuggest that corals with low growth rates (µCa/mgN/h)and high metabolic rates (µO₂/cm²/hr), such as the massivespecies, acclimatize more effectively than those with high growthrates and low metabolic rates, a feature that is characteristicof branching species. We conclude that studies of protein turnover,combined with temporally relevant investigations into the dynamicaspects of coral dinoflagellate symbioses will provide considerableinsight into why corals exhibit such a high level of variationin response to the same environmental challenge. Furthermore,a more detailed understanding of acclimatization mechanismsis essential if we are to predict how a coral assemblage willrespond to present and future environmental challenges.     

Long-term survival despite low genetic diversity in the critically endangered Madagascar fish-eagle

The critically endangered Madagascar fish-eagle ( Haliaeetus vociferoides ) is considered to be one of the rarest birds of prey globally and at significant risk of extinction. In the most recent census, only 222 adult individuals were recorded with an estimated total breeding population of no more than 100–120 pairs. Here, levels of Madagascar fish-eagle population genetic diversity based on 47 microsatellite loci were compared with its sister species, the African fish-eagle ( Haliaeetus vocifer ), and 16 of these loci were also characterized in the white-tailed eagle ( Haliaeetus albicilla ) and the bald eagle ( Haliaeetus leucocephalus ). Overall, extremely low genetic diversity was observed in the Madagascar fish-eagle compared to other surveyed Haliaeetus species. Determining whether this low diversity is the result of a recent bottleneck or a more historic event has important implications for their conservation. Using a Bayesian coalescent-based method, we show that Madagascar fish-eagles have maintained a small effective population size for hundreds to thousands of years and that its low level of neutral genetic diversity is not the result of a recent bottleneck. Therefore, efforts made to prevent Madagascar fish-eagle extinction should place high priority on maintenance of habitat requirements and reducing direct and indirect human persecution. Given the current rate of deforestation in Madagascar, we further recommend that the population be expanded to occupy a larger geographical distribution. This will help the population persist when exposed to stochastic factors (e.g. climate and disease) that may threaten a species consisting of only 200 adult individuals while inhabiting a rapidly changing landscape.     

Molecular Characterization of the Mouse Tyrosinase Gene:Pigment Cell-Specific Expression in Transgenic Mice

Tyrosinase is the key enzyme in melanin synthesis, and is expressed in the pigment epithelium of the retina, a cell layer derived from the optic cup; and in neural crest-derived melanocytes of skin, hair follicle, choroid, and iris. The tyrosinase gene has been cloned and shown to map to the well-characterized c-locus (albino locus) of the mouse. Subsequent studies demonstrated that a functional tyrosinase minigene was able to rescue the albino phenotype in transgenic mice. The transgene was expressed in a cell type-specific manner in skin and eye. During development of the mouse, the tyrosinase gene is expressed in the pigment epithelium of the retina as early as day 10.5 of gestation. In the hair follicle, tyrosinase gene expression is detected from day 16.5 onwards. This cell-type–specific expression is largely reproduced in transgenic mice. Our results suggest that sequences in the immediate vicinity of the mouse tyrosinase gene are sufficient to provide cell type-specificity and developmental regulation in melanocytes and the pigment epithelium.