International Society for Reef Studies

International Society for Reef Studies     

Cell-substrate adhesion and metastatic potential of cultured mesothelioma cells induced by asbestos

Cell-substrate adhesion was quantified for two cultured mesothelioma cell lines (epitheliomatus and sarcomatous) on glass, fibronectin and laminin substrates. Interference reflection microscopy (IRM) was used to image the adhesion patterns of cells and a grey level analysis was employed to quantify adhesion. Sarcomatous cells demonstrated marked adhesion to glass and fibronectin-coated substrates but not to laminin-coated substrate, with the greatest adhesion occurring on the fibronectin-coated surface. This adhesion was accompanied by cytoplasmic spreading. By contrast, epitheliomatous cells showed little tendency to adhere to any of the substrates and only showed significant spreading when in contact with the laminin substrate (P < 0.01). A bioassay was used to determine the metastatic potential of each of the cell lines. Via the intravenous route, the sarcomatous cells killed the host rats in 24.7 ± 1.5 (S.D.) days compared to 27.3 ± 0.9 (S.D.) days for the epitheliomatous cells (P < 0.01). After subcutaneous inoculation of tumour cells, the sarcomatous cells killed the host rats in 54.7 ± 0.7 (S.D.) days compared to 48.5 ± 0.5 (S.D.) days for the epitheliomatous cells (P < 0.01). We conclude that the results of the metastasis bioassays were consistent with the predicted behavior of these cell lines based on their ability to adhere to substrates in the in vitro adhesion assays.     

Retrotransposon-induced ectopic expression of the Om(2D) gene causes the eye-specific Om(M) phenotype in Drosophila ananassae

Optic morphology (Om) mutations in Drosophila ananassae map to at least 22 loci, which are scattered throughout the genome. Om mutations are all semidominant, neomorphic, nonpleiotropic, and associated with the insertion of a retrotransposon, tom. We have found that the Om(2D) gene encodes a novel protein containing histidine/proline repeats, and is ubiquitously expressed during embryogenesis. The Om(2D) RNA is not detected in wild-type eye imaginal discs, but is abundantly found in the center of the eye discs of Om(2D) mutants, where excessive cell death occurs. D. melanogaster flies transformed with the Om(2D) cDNA under control of the hsp70 promoter display abnormal eye morphology when heat-shocked at the third larval instar stage. These results suggest that the Om(2D) gene is not normally expressed in the eye imaginal discs, but its ectopic expression, induced by the tom element, in the eye disc of third instar larvae results in defects in adult eye morphology.     

Morphological and proliferative responses of endothelial cells to hydrostatic pressure: Role of fibroblast growth factor

Subconfluent bovine pulmonary artery endothelial cells on rigid substrates were exposed to 1.5–15 cm H₂O sustained hydrostatic pressure for up to 7 days and exhibited elongation, cytoskeletal rearrangement, increased cell proliferation, and bilayering. The role of basic fibroblast growth factor (bFGF) in the mechanism(s) of these endothelial cell responses to sustained hydrostatic pressure was investigated. Evidence that bFGF was released from endothelial cells exposed to sustained hydrostatic pressure or compression was provided by the following experimental results: (1) Cells exposed to control (3 mm H₂O) pressure displayed intense nuclear and cytoplasmic bFGF staining by immunocytochemical techniques; this staining was absent in cells exposed to 10 cm H₂O for 7 days. (2) Conditioned medium from endothelial cells exposed to 10 cm H₂O for 7 days contained at ansferable, growth-promoting activity exhibiting heparin-Sepharose affinity, lability to both heat and freeze/thawing, and neutralization by anti-bovine bFGF. (3) Suramin (0.1 mM), a growth-factor receptor inhibitor, abrogated the proliferative and morphological responses of endothelial cells exposed to sustained hydrostatic pressure. Endothelial cells exposed to elevated hydrostatic pressure demonstrated no detectable decrement in cell viability as assessed by Trypan blue exclusion. The results of the present study indicate that hydrostatic pressure or compression can induce bFGF release from endothelial cells independent of cell injury or death; bFGF is subsequently responsible for the morphological, proliferative, and bilayering responses of endothelial cells to hydrostatic pressure. © 1993 Wiley-Liss, Inc.     

Mechanism of interferon action: evidence for intermolecular autophosphorylation and autoactivation of the interferon-induced, RNA-dependent protein kinase PKR.

The interferon-induced RNA-dependent protein kinase (PKR) is postulated to have an important regulatory role in the synthesis of viral and cellular proteins. Activation of the enzyme requires the presence of a suitable activator RNA and is accompanied by an autophosphorylation of PKR. Active PKR phosphorylates the alpha subunit of protein synthesis eukaryotic initiation factor 2, resulting in an inhibition of translation initiation. The mechanism of autophosphorylation is not well understood. Here we present evidence that the autophosphorylation of human PKR can involve intermolecular phosphorylation events, i.e., one PKR protein molecule phosphorylating a second PKR molecule. Both wild-type PKR and the point mutant PKR(K296R) synthesized in vitro were phosphorylated, even though PKR(K296R) was deficient in kinase catalytic activity. Phosphorylation of both wild-type PKR and PKR(K296R) was inhibited in the presence of 2-aminopurine. Furthermore, purified human recombinant PKR(K296R) was a substrate for the purified wild-type human PKR kinase. This intermolecular phosphorylation of mutant PKR(K296R) by wild-type PKR was dependent on double-stranded RNA and was inhibited by 2-aminopurine. Finally, PKR mRNA was capable of mediating an autoactivation of wild-type PKR kinase autophosphorylation in vitro.     

Alkalinity Tolerance of Woody Species Used in Bauxite Waste Rehabilitation, Western Australia

Glasshouse trials, using trickle irrigation and increasing levels of NaOH-induced alkalinity, identified species that could be expected to tolerate the high-pH conditions of bauxite processing waste residue sites. Of 29 taxa tested, the most tolerant were Casuarina obesa, Melaleuca lanceolata, M. armillaris, M. nesophila, Eucalyptus loxophleba, E. halophila, E. platypus, Tamarix aphylla, and a particular clone of E. camaldulensis; E. spathulata, E. tetragona, E. preissiana, E. gomphocephala, E. diptera, and E. occidentalis proved to be relatively sensitive to severe alkaline conditions. Tolerance appeared to relate to an ability to maintain root membrane function, nutrient uptake balance, and ultimately root tissue structure while under increasing levels of alkalinity stress. Species normally inhabiting alkaline soils tended to have increased growth rates in nutrient irrigation conditions between pH 8 and 10 compared with control plants irrigated with nutrient solutions of pH values near 7.4. However, once the irrigation solutions reached pH 12 and the buffering capacity of the soil appeared to be exceeded, the condition of susceptible plants rapidly declined and death followed. Sensitive plants initially showed symptoms related to nutrient deficiency, followed by wilting and death as the root systems failed. Field trial conditions in the bauxite residue impoundments at Kwinana, Western Australia, include soils with pH values as high as 11.00. In general, the relative survival and growth of seedlings after eight months were predicted by the response under glasshouse trial conditions. Appropriately designed stress trials can be important ecological techniques in choosing species most capable of surviving difficult environmental conditions in the rehabilitation of damaged landscapes.     

Serum concentrations of reproductively-related circulating steroid hormones in the free-ranging lemon shark,Negaprion brevirostris

Synopsis We have examined the concentrations of reproductively-related steroid hormones in 5 species of carcharhinid sharks, marine fishes possessing the unique attribute of placental viviparity. Measurements of serum estradiol, testosterone, progesterone, dihydrotestosterone, and corticosterone have provided baseline data for these hormones in both immature and adult male and female placental sharks. Our studies include: (1) changes in hormonal levels during maturation, (2) concentrations of circulating steroid hormones during peak breeding season, and (3) hormonal levels during gestation, including the collection of serial samples through and beyond birth from free-ranging lemon sharks. Our data suggest that steroids, important in regulating reproduction in higher mammals, are also essential in these cartilaginous fishes.     

Mortality rates of desert vegetation during high-intensity drought at Uluru-Kata Tjuta National Park,Central Australia

Precipitation variability and heatwaves are expected to intensify over much of inland Australia under most projected climate change scenarios. This will undoubtedly have impacts on the biota of Australian dryland systems. However, accurate modelling of these impacts is presently impeded by a lack of empirical research on drought/heatwave effects on native arid flora and fauna. During the 2018–2021 Australian drought, many parts of the continent's inland experienced their hottest, driest period on record. Here, we present the results of a field survey in 2021 involving indigenous rangers, scientists and national parks staff who assessed plant dieback during this drought at Ulur u-Kata Tjut a National Park (UKTNP), central Australia. Spatially randomized quadrat sampling of eight common and culturally important plants indicated the following plant death rates across UKTNP (in order of drought susceptibility): desert myrtle (Aluta maisonneuvei subsp. maisonneuvei) (91%), yellow flame grevillea (Grevillea eriostachya) (79%), Maitland's wattle (Acacia maitlandii) (67%), waxy wattle (A. melleodora) (65%), soft spinifex grass (Triodia pungens) (53%), mulga (A. aneura) (42%), desert oak (Allocasuarina decaisneana) (22%) and quandong (Santalum acuminatum) (0%). The sampling also detected that seedling recruitment was absent or minimal for all plants except soft spinifex, while a generalized linear mixed model (GLMM) indicated two-way interactions among species, plant size and stand density as important predictors of drought survival of adult plants. A substantial loss of biodiversity has occurred at UKTNP during the recent drought, with likely drivers of widespread plant mortality being extreme multi-year rainfall deficit (2019 recorded the lowest-ever annual rainfall at UKTNP [27 mm]) and record high summer temperatures (December 2019 recorded the highest-ever temperature [47.1°C]). Our findings indicate that widespread plant death and extensive vegetation restructuring will occur across arid Australia if the severity and frequency of droughts increase under climate change.     

Ecological and behavioral consequences of digestion in frugivorous animals

The postingestional effect of seed size and mass and nutrient composition on fruit profitability are reviewed. It is emphasized that profitability results from the interaction between fruit characteristics and the physiological and morphological traits of frugivores. The processes by which frugivores regurgitate or defecate seeds and the differential processing of the nutrient rich pulp and ballast in fruit are strongly dependent on the interaction between frugivore gut morphology and seed size. Euphonias that lack a functional gizzard defecate seeds, whereas tanagers that have a delete a functional gizzards regurgitate seeds. Some frugivores separate seeds from pulp and exocarp in the gizzard. It is hypothesized that the gizzard plays an important role in determining the postigestional fate of different pulp components. Although fruit nutrient content is often invoked as a determinant of frugivore feeding choices, studies that rely on proximal nutrient analysis, have often failed to find clear nutrient composition-preference correlations. It is argued that a partial reason for this failure is that proximal nutrient analysis ignores the complexities of fruit digestion. The ecological and physiological correlates of lipid and sugar assimilation are used to identify the limitations of traditional proximal nutrient analyses in fruit-frugivore studies. We suggest that recognizing the intricacies of the digestive characteristics of frugivores may reveal a much richer patterning in the interaction of frugivores with plants than has been previously hypothesized.