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We evaluated rat bone viability using a bone viability index (BVI). To evaluate hypothermic ischaemic bone injury, 21 amputated hind limbs of Fischer rats were preserved at hypothermia (4°C) for 1, 3 and 6 hours. To evaluate hypothermic ischaemia/reperfusion injury, another 28 amputated limbs were transplanted to recipient rats after hypothermic ischaemia for 3 and 6 hours, respectively. Total RNA isolated from each tibia was fractionated by electrophoresis and hybridised with ³²P-labelled cDNA of GAPDH, and the radioactivity of intact and degraded GAPDH mRNA measured. BVI was calculated as follows, BVI = {A / (A + B)} × 100, where A and B represent the radioactivities corresponding to intact and degraded GAPDH mRNA bands, respectively. In the hypothermic ischaemic insult group, BVIs were comparable to those of controls. However, in the 3-hour hypothermic ischaemia/reperfusion group, BVI was lower than that of the controls. Likewise, there was a significant difference between the 6-hour ischaemia/reperfusion group and controls. These results showed that bone viability decreased even after just a 3-hour hypothermic ischaemia/reperfusion.

The low-intensity ultrasound effects on MCF7 (human breast adenocarcinoma) and MCF10A (healthy breast cells) have been investigated at different sonication protocol to probe the effectiveness and the selectivity of the ultrasound (US) treatment and to understand the implications between cell mortality, biomechanical interactions and cell elastic modulus. Experiments performed at fixed and variable frequency demonstrated the effectiveness of some protocols in killing carcinogenic cells and the healthy cells insensitivity. Variation of elastic properties of MCF7 cells exposed to US under varying sonication conditions was examined. Sonication was carried out at fixed frequency (as it is usually done in therapy protocols), between 400kHz and 620kHz, following two protocols: (i) at fixed power output; (ii) at fixed voltage of the US generator. Evolution of cell stiffness during the US treatment was monitored via atomic force spectroscopy (AFS). It was found that cell mortality has a similar trend of variation with respect to sonication frequency regardless of the way specimens are exposed to US. Mechanical properties do not show a uniform trend with respect to frequency, but variations of Young’s modulus are more marked near the very low (400–480) kHz or very high frequencies (580–620) kHz. The observed variations may be related to mechanical interactions occurring in the cell culture, suggesting a primacy of the environment on other factors.

Developmental time is a trait of great relevance to fitness in all organisms. In holometabolous species that occupy ephemeral habitat, like Drosophila melanogaster, the impact of developmental time upon fitness is further exaggerated. We explored the trade-offs surrounding developmental time by selecting 10 independent populations from two distantly related selection treatments (CB_1-5 and CO_1-5) for faster development. After 125 generations, the resulting accelerated populations (ACB_1-5 and ACO_1-5) displayed net selection responses for development time of -33.4 hours (or 15%) for ACB and -38.6 hours (or 17%) for ACO. Since most of the change in egg-to-adult developmental time was accounted for by changes in larval duration, the “accelerated” larvae were estimated to develop 25-30% faster than their control/ancestor populations. The responses of ACB and ACO lines were remarkably parallel, despite being founded from populations evolved independently for more than 300 generations. On average, these “A” populations developed from egg to adult in less than eight days and produced fertile eggs less than 24 hours after emerging. Accelerated populations showed no change in larval feeding rate, but a reduction in pupation height, the latter being a trait relating to larval energetic expenditure in wandering prior to pupation. This experiment demonstrates the existence of a negative evolutionary correlation between preadult developmental time and viability, as accelerated populations experienced a severe cost in preadult survivorship. In the final assay generation, viability of accelerated treatments had declined by more than 10%, on average. A diallel cross demonstrated that the loss of viability in the ACO lines was not due to inbreeding depression. These results suggest the existence of a rapid development syndrome, in which the fitness benefits of fast development are balanced by fitness costs resulting from reduced preadult survivorship, marginal larval storage of metabolites, and reduced adult size.

Ralstonia solanacearum biovar (bv) 2 causes bacterial wilt and potato brown rot in solanaceous plants in temperate areas. The pathogen is able to persist in the environment, where it is frequently disseminated by watercourses. In these habitats, the effect of long-term oligotrophy on its survival remains to be ascertained. On that purpose river water microcosms were inoculated with R. solanacearum bv 2, incubated at 24°C and monitored for total, viable and culturable bacterial cell numbers up to four years. Within the first year, R. solanacearum bv 2 populations remained roughly constant in the initial levels. Then and until the fourth year, total counts slowly increased, while viability slightly declined and culturability decreased to a greater extent, pointing out a proportion of the bacterial populations entering the viable but non-culturable state. As cells in this state are not detected by cultivation-based methods, they represent a new challenge in designing strategies for control of the bacterial wilt disease.

Doxorubicin (DOXO), an anthracycline antibiotic, is a potent anticancer agent with severe toxic side-effects that have been attributed to its ability to generate free radicals (ROS), particularly in mitochondria. In this study, a prokaryotic organism, Salmonella typhimurium, was used to evaluate the effect of the drug on ROS-scavenging enzymes, catalase (CAT) and superoxide dismutase (SOD) activity, and on cell growth and viability. Increasing DOXO concentrations in the culture medium led to increasingly prolonged lag period, reduced growth rate and cell viability, and inhibition of SOD and CAT activity. SOD activity dropped to 66% of the control value in cells grown with 1 µg/ml DOXO and to 34% and 28% in cells grown with 150 and 300 µg/ml DOXO. CAT activity decreased progressively from 93% in 1 µg/ml DOXO to 30% in 150 µg/ml DOXO and was undetectable in 300 µg/ml. Growth in the presence of DOXO led to alterations in cytokinesis, as seen upon examination of cells under fluorescence microscope.