Levels and probable origin of predatory scarring on humpback whales (Megaptera novaeangliae) in east Australian waters

To investigate the incidence of non-lethal predation in Southern Hemisphere whales, more than 3400 fluke-identification photographs from resight histories of 1436 east Australian humpback whales were examined for evidence of predatory markings. Photographs were obtained from 1984 to 1996 at various locations along the east coast of Australia, from northern Queensland to southern New South Wales. Photographs were classified in terms of the level and type of scarring. The possible predator and whether the markings appeared fresh were also noted. In all, 17% of identified east Australian humpbacks possessed some form of predatory scarring, 57% of which was minor and 43% major. Almost all predatory scarring was consistent with that inflicted by killer whales. Only three whales demonstrated an increase in the level of predatory scarring after their first sightings. Two incidents of fresh scarring were recorded, and one fatal killer whale attack on a humpback whale calf was directly observed. The overall level of predatory scarring found in this study is comparable to those found in studies for Northern Hemisphere humpback whales. The low incidence of adult whales showing their first sign of predatory scarring after their initial sighting, and the small number possessing recent scarring, support the idea that east Australian humpback whales experience most predatory attacks early in life.

Field trial of a new aeration system for enhancing biodegradation in a biopile

The influence of a new aeration system on the biopile performance was investigated. The purpose was to increase biodegradation efficiency by optimising airflow through the pile. During a 1-month field trial, the performance of a new system using two perforated vertical pipes with wind-driven turbines was compared with that of a standard pile configuration with two horizontal perforated pipes. Both piles were composed of a similar mix of diesel-contaminated soils, woodchips, compost and NPK fertiliser. Hydrocarbons were recovered using solvent extraction, and determined both gravimetrically and by gas chromatography. Total heterotrophs, pH and moisture content were also assessed. Air pressure measurements were made to compare the efficiency of suction in the pipes. Results at the end of the experiment showed that there was no significant difference between the two piles in the total amount of hydrocarbon biodegradation. The normalised degradation rate was, however, considerably higher in the new system than in the standard one, suggesting that the vertical venting method may have improved the efficiency of the biological reactions in the pile. The pressure measurements showed a significant improvement in the suction produced by the new aeration system. However, many factors other than the airflow (oxygen supply) may influence and limit the biodegradation rates, including moisture content, age of contaminants and the climatic conditions. Additional experiments and modelling need to be carried out to explore further the new aeration method and to develop criteria and guidelines for engineering design of optimal aeration schemes in order to achieve maximum biodegradation in biopiles. (C) 2003 Elsevier Ltd. All rights reserved.

Physiological thermoregulation in a crustacean? Heart rate hysteresis in the freshwater crayfish Cherax destructor

Differential heart rates during heating and cooling (heart rate hysteresis) are an important thermoregulatory mechanism in ectothermic reptiles. We speculate that heart rate hysteresis has evolved alongside vascularisation, and to determine whether this phenomenon occurs in a lineage with vascularised circulatory systems that is phylogenetically distant from reptiles, we measured the response of heart rate to convective heat transfer in the Australian freshwater crayfish, Cherax destructor. Heart rate during convective heating (from 20 to 30 degreesC) was significantly faster than during cooling for any given body temperature. Heart rate declined rapidly immediately following the removal of the heat source, despite only negligible losses in body temperature. This heart rate 'hysteresis' is similar to the pattern reported in many reptiles and, by varying peripheral blood flow, it is presumed to confer thermoregulatory benefits particularly given the thermal sensitivity of many physiological rate functions in crustaceans. (C) 2004 Published by Elsevier Inc.

Wolbachia-induced cytoplasmic incompatibility as a means for insect pest population control

Biological control is the purposeful introduction of parasites, predators, and pathogens to reduce or suppress pest populations. Wolbachia are inherited bacteria of arthropods that have recently attracted attention for their potential as new biocontrol agents. Wolbachia manipulate host reproduction by using several strategies, one of which is cytoplasmic incompatibility (CI) [Stouthamer, R., Breeuwer, J. A. J. & Hurst, G. D. D. (1999) Annu. Rev. Microbiol. 53,71-102]. We established Wolbachia-infected lines of the medfly Ceratitis capitata using the infected cherry fruit fly Rhagoletis cerasi as donor. Wolbachia induced complete CI in the novel host. Laboratory cage populations were completely suppressed by single releases of infected males, suggesting that Wolbachia-induced CI could be used as a novel environmentally friendly tool for the control of medfly populations. The results also encourage the introduction of Wolbachia into pest and vector species of economic and hygenic relevance to suppress or modify natural populations.

Divergent environments and population bottlenecks fail to generate premating isolation in Drosophila pseudoobscura

While the feasibility of bottleneck-induced speciation is in doubt, population bottlenecks may still affect the speciation process by interacting with divergent selection. To explore this possibility, I conducted a laboratory speciation experiment using Drosophila pseudoobscura involving 78 replicate populations assigned in a two-way factorial design to both bottleneck (present vs. absent) and environment (ancestral vs. novel) treatments. Populations independently evolved under these treatments and were then tested for assortative mating and male mating success against their common ancestor. Bottlenecks alone did not generate any premating isolation, despite an experimental design that was conducive to bottleneck-induced speciation. Premating isolation also did not evolve in the novel environment treatment, neither in the presence nor absence of bottlenecks. However, male mating success was significantly reduced in the novel environment treatment, both as a plastic response to this environment and as a result of environment-dependent inbreeding effects in the bottlenecked populations. Reduced mating success of derived males will hamper speciation by enhancing the mating success of immigrant, ancestral males. Novel environments are generally thought to promote ecological speciation by generating divergent natural selection. In the current experiment, however, the novel environment did not cause the evolution of any premating isolation and it reduced the likelihood of speciation through its effects on male mating success.

Environmentalizing economic development: a South Asian perspective

South Asia's pursuit of economic development has entailed considerable damage to and exposed the fragility of the physical environment of the region. This paper provides an analytical overview of the of the environmental problem that manifest themselves in South Asia in a comparative perspective with East and Southeast Asian countries as well as selected developed market economics. To date, South Asian development process has been environment-intensive and environment-depleting. It is argued that environmental problems are likely to set serious constraints to sustain growth in production to support a growing population. By exploring the relationship between indices of human welfare and bio-diversity conservation. the paper exposes the dichotomy of the development process. Finally, the study underscores the need for a range of policy options that rely both based and non-market based instruments in an integrated setting to enviromnentalize South Asian economic development. (C) 2004 Elsevier B.V All rights reserved.

Quantitative description of the interaction between folate and the folate-binding protein from cow's milk

A detailed study has been carried out on the dependence of folate binding on the concentration of FBP (folate-binding protein) at pH 5.0, conditions selected to prevent complications arising from the pre-existing self-association of the acceptor. In contrast with the mandatory requirement that reversible interaction of ligand with a single acceptor site should exhibit a unique, rectangular hyperbolic binding curve, results obtained by ultrafiltration for the FBP-folate system required description in terms of (i) a sigmoidal relationship between concentrations of bound and free folate and (ii) an inverse dependence of affinity on FBP concentration. These findings have been attributed to the difficulties in determining the free ligand concentration in the FBP-folate mixtures for which reaction is essentially stoichiometric. This explanation also accounts for the similar published behaviour of the FBP-folate system at neutral pH, which had been attributed erroneously to acceptor self-association, a phenomenon incompatible with the experimental findings because of its prediction of a greater affinity for folate with increasing FBP concentration.

The inhibitory effect of pentobarbitone on reverse transcription-PCR

Pentobarbitone sodium (Sodium 5-ethyl-5[1-methylbutyl]-pentobarbitone) is a short-acting barbiturate that is commonly used to euthanase animals. As part of our studies into the molecular genetics of copper toxicosis in Bedlington terrier dogs, reverse-transcription (RT)-PCR was noted to always fail on RNA samples collected from livers of dogs sacrificed by pentobarbitone injection. When samples were collected without pentobarbitone, however, RTPCR was always successful. We suspected the possible inhibition by pentobarbitone sodium of either reverse transcriptase or Taq polymerase. In vitro studies showed that pentobarbitone interference of PCR occurred at >4 mug/mul. To identify if pentobarbitone produced competitive inhibition, each components (Taq polymerase, MgCl2, dNTP, etc.) of the PCR was individually altered. However, inhibition still persisted, suggesting that multiple PCR components may be affected. Also it was shown that pentobarbitone interference was not dependent on the PCR product size. Simple dilution of pentobarbitone contaminated DNA solutions, and the addition of bovine serum albumin (BSA) to the PCR mix overcame pentobarbitone interference. In vivo, PCR by pentobarbitone was found to be compounded by high DNA concentration and pentobarbitone contamination. In addition, both high DNA concentration and pentobarbitone contamination could be overcome through dilution and the addition of BSA. Further work is required to quantify pentobarbitone concentration in the liver-extracted DNA and RNA samples before this inhibition effect on PCR can be fully elucidated. (C) 2004 Elsevier B.V. All rights reserved.

Ratiometric and nonratiometric Ca2+ indicators for the assessment of intracellular free Ca2+ in a breast cancer cell line using a fluorescence microplate reader

Transporters of Ca2+ are potential drug targets and Ca2+ is a useful signal in the assessment of G-protein-coupled receptor activation. Assays involving the assessment of intracellular Ca2+ using microplate readers most often use Ca2+ indicators which do not exhibit a spectra shift on Ca2+ binding (e.g. fluo-3). Indicators that do exhibit a spectral shift upon Ca2+ binding (e.g. fura-2) offer potential advantages for the calibration of intracellular Ca2+ levels. However, experimental limitations may limit the use of ratiometric dyes in microplate readers capable of screening. In this study, we compared the assessment of intracellular Ca2+ in adherent breast cancer cells using ratiometric and nonratiometric Ca2+ indicators. Our results demonstrate that both fluo-3 and fura-2 detect ATP dose-dependent increases in intracellular Ca2+ in the MCF-7 breast cancer cell line and that some of the limitations in the use of fura-2 appear to be overcome by the use of glass bottom microplates. The calibrated intracellular Ca2+ levels derived using fura-2 are consistent with those from microscopy and cuvette-based studies. Fura-2 may be useful in microplate studies, where cell lines with different properties are compared or where screening treatments lead to differences in the number of cells or dye loading. (C) 2003 Elsevier B.V. All rights reserved.

Searching oligo sets of human chromosome 12 using evolutionary strategies

DNA Microarray is a powerful tool to measure the level of a mixed population of nucleic acids at one time, which has great impact in many aspects of life sciences research. In order to distinguish nucleic acids with very similar composition by hybridization, it is necessary to design microarray probes with high specificities and sensitivities. Highly specific probes correspond to probes having unique DNA sequences; whereas highly sensitive probes correspond to those with melting temperature within a desired range and having no secondary structure. The selection of these probes from a set of functional DNA sequences (exons) constitutes a computationally expensive discrete non-linear search problem. We delegate the search task to a simple yet effective Evolution Strategy algorithm. The computational efficiency is also greatly improved by making use of an available bioinformatics tool.

Moving molecules: mRNA trafficking in mammalian oligodendrocytes and neurons

Numerous mRNA molecules are localized in regions of the dendrites of neurons, some moving along dendrites in response to synaptic activity. The proteins encoded by these RNAs have diverse functions, including participation in memory formation and long-term potentiation. Recent experiments have shown that a cytoplasmic RNA trafficking pathway described for oligodendrocytes also operates in neurons. Transported RNAs possess a cis-acting element that directs them to granules, which are transported along microtubules by the motor proteins kinesin and dynein. These RNA molecules are recruited to the cytoplasmic transport granules by cooperative interaction with a cognate trans-acting factor. mRNAs containing the 11-nucleotide A2RE11 or 21-nucleotide A2RE sequences bind heterogeneous nuclear ribonucleoproteins A2 and A3, which are abundant in the brain. Mutations in this cis-acting element that weaken its interaction with hnRNP A2 also interfere with RNA trafficking. Several dendritically localized mRNAs, including those encoding calcium-calmodulin-dependent protein kinase 11 a subunit and neurogranin, possess A2RE-like sequences, suggesting that they may be localized by interaction with these heterogeneous nuclear ribonucleoproteins. Calcium-calmodulin-dependent protein kinase 11 a subunit is of particular interest: Its RNA is transported in depolarized neurons, and the protein it encodes is essential for establishing long-term memory. Several other cis-acting sequences and trans-acting factors that participate in neuronal RNA localization have been discovered.

Increased expression of the MBP mRNA binding protein HnRNP A2 during oligodendrocyte differentiation

Heterogeneous nuclear ribonucleoprotein (hnRNP) A2, a trans-acting factor that mediates intracellular trafficking of myelin basic protein (MBP) mRNA to the myelin compartment in oligodendrocytes, is most abundant in the nucleus, but shuttles between the nucleus and cytoplasm. In the cytoplasm, it is associated with granules that transport mRNA from the cell body to the processes of oligodendrocytes. We found that the overall level of hnRNP A2 increased in oligodendrocytes as they differentiated into MBIP-positive cells, and that this augmentation was reflected primarily in the cytoplasmic pool of hnRNP A2 present in the form of granules. The extranuclear distribution of hnRNP A2 was also observed in brain during the period of myelination in vivo. Methylation and phosphorylation have been implicated previously in the nuclear to cytoplasmic distribution of hnRNPs, so we used drugs that block methylation and phosphorylation of hnRNPs to assess their effect on hnRNP A2 distribution and mRNA trafficking. Cultures treated with adenosine dialdehyde (AdOx), an inhibitor of S-adenosyl-L-homocysteine hydrolase, or with 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB), a drug that inhibits casein kinase 2 (CK2), maintained the preferential nuclear distribution of hnRNP A2. Treatment with either drug affected the transport of RNA trafficking granules that remained confined to the cell body. (C) 2004 Wiley-Liss, Inc.