Tributyltin causes cytochrome C release from isolated mitochondria by two discrete mechanisms

Using isolated liver mitochondria we show that low concentrations of TBT (0.5 microM) cause the release of mitochondrial cytochrome c, in the presence of Ca(2+). This is reflected in a rapid loss of membrane potential (DeltaPsi(m)), and a large-amplitude swelling characteristic of mitochondrial permeability transition (MPT). Despite this, the inclusion of cyclosporin A could not prevent the release of cytochrome c. Further, in the absence of Ca(2+), low concentrations of TBT (0.5 microM) resulted in a slow sub-maximal shift of DeltaPsi(m), not characteristic of MPT, which was still paralleled by a release of cytochrome c. Further experiments showed that the loss of DeltaPsi(m) in the absence of Ca(2+) was due to a combination of inhibition of respiration and a direct uncoupling effect on the respiratory chain. Under these conditions, rapid swelling of mitochondria could be demonstrated, due to chloride exchange over the inner mitochondrial membrane. Taken together these data suggest that TBT can induce the release of cytochrome c in intact cells by at least two mechanisms. The first and critical mechanism is initiated immediately the mitochondria sense the presence of TBT and involves a slow loss of DeltaPsi(m) and induction of swelling, which allows release of cytochrome c in a relatively non-specific manner and independently from a rise in [Ca(2+)](i). The second mechanism involves the induction of formal MPT as intracellular [Ca(2+)](i) increases. These data help to explain previous observations in intact lymphocytes demonstrating TBT-induced release of mitochondrial cytochrome c in the absence of a rise in [Ca(2+)](i) (Stridh, H., Gigliotti, D., Orrenius, S., and Cotgreave, I. A. (1999) Biochem. Biophys. Res. Commun. 266, 460-465).     

On the dynamics of the stocks of blue mussels (Mytilus edulis L.) in the Danish Wadden Sea

As biological basis for the monitoring programme for the commercially exploited stock(s) of mussels (Mytilus edulis L.) in the Danish Wadden Sea, samples of mussels have been collected regularly since 1986, both from sub-tidal and inter-tidal mussel beds. These samples are the basis for the estimation of total biomass. They also provide data on size frequency distributions, which have been analysed for cohort identification resulting in length at age data, which again have been used for estimating parameters (L and K) for the von Bertalanffy growth equation (VBGE) as well as mortality parameters. By applying these in the Beverton & Holt model, estimates of average biomass and annual production (P) of the mussels have been obtained together with possible fisheries yields from the beds. The growth and mortality parameters and the figures for annual production and P/B are compared with figures from other investigations. These analyses have been the basis for annual assessments of the mussel stocks, which again are used in the current management of mussel fishery in the Danish Wadden Sea.     

Shake-up and shake-off excitations with associated electron losses in X-ray studies of proteins

Photoionization of an atom by X-rays usually removes an inner shell electron from the atom, leaving behind a perturbed "hollow ion" whose relaxation may take different routes. In light elements, emission of an Auger electron is common. However, the energy and the total number of electrons released from the atom may be modulated by shake-up and shake-off effects. When the inner shell electron leaves, the outer shell electrons may find themselves in a state that is not an eigen-state of the atom in its surroundings. The resulting collective excitation is called shake-up. If this process also involves the release of low energy electrons from the outer shell, then the process is called shake-off. It is not clear how significant shake-up and shake-off contributions are to the overall ionization of biological materials like proteins. In particular, the interaction between the outgoing electron and the remaining system depends on the chemical environment of the atom, which can be studied by quantum chemical methods. Here we present calculations on model compounds to represent the most common chemical environments in proteins. The results show that the shake-up and shake-off processes affect approximately 20% of all emissions from nitrogen, 30% from carbon, 40% from oxygen, and 23% from sulfur. Triple and higher ionizations are rare for carbon, nitrogen, and oxygen, but are frequent for sulfur. The findings are relevant to the design of biological experiments at emerging X-ray free-electron lasers.     

Columnar denitrification of water by immobilized Pseudomonas denitrificans cells

Summary Whole cells of Pseudomonas denitrificans, immobilized in alginate gel, were used for columnar denitrification of ground water. Ethanol was selected as a suitable carbon source and the C/N-ratio necessary for satisfactory nitrate reduction was established (1.6 mg ethanol-C/mg nitrate-N). The course of the reaction and the diffusional limitations were investigated during columnar denitrification. The mechanical integrity of the gel matrix, as judged from leakage of cells was studied. The release of cells into the effluent was effectively inhibited (<10² cells/ml) by the use of different filter devices. The operational characteristics were determined by studying a column operating for nearly four months. Theoretically, the alginate gel column should, from high nitrate drinking water (22 mg NO ₃ ^– -N/1), produce 3 1 of denitrified water/kg gel/h (wet wt.) during a period of two months. The regeneration of nitrate reduction activity by means of activation in nutrient media proved a useful tool for restoring initial activity, the gel column having shown no loss in activity at the end of the operation period.     

Cell death mechanisms: Cross-talk and role in disease

Multiple cell death mechanisms operate in both uni- and multicellular organisms. Hence, research during the past forty years has revealed that apoptosis is not the only cell death program involved in the regulation of tissue homeostasis and the removal of unwanted cells in biological organisms. While the molecular pathways of apoptosis and necrosis are now relatively well established, the precise mechanisms of other cell death modalities, and their cross-talk, require additional study. This is particularly important, since many human disorders can be attributed, directly or indirectly, to defective cell death mechanisms. In this review we shall discuss the characteristics and cross-talk between various modes of cell death and their role in cell death-related disorders, notably, neurodegenerative disease and cancer.     

Multiplying steady-state culture in multi-reactor system

Cultivation of microorganisms in batch experiments is fast and economical but the conditions therein change constantly, rendering quantitative data interpretation difficult. By using chemostat with controlled environmental conditions the physiological state of microorganisms is fixed; however, the unavoidable stabilization phase makes continuous methods resource consuming. Material can be spared by using micro scale devices, which however have limited analysis and process control capabilities. Described herein are a method and a system combining the high throughput of batch with the controlled environment of continuous cultivations. Microorganisms were prepared in one bioreactor followed by culture distribution into a network of bioreactors and continuation of independent steady state experiments therein. Accelerostat cultivation with statistical analysis of growth parameters demonstrated non-compromised physiological state following distribution, thus the method effectively multiplied steady state culture of microorganisms. The theoretical efficiency of the system was evaluated in inhibitory compound analysis using repeated chemostat to chemostat transfers.     

Population structure and dynamics of wood mouse Apodemus sylvaticus in Iceland

Five-year-studies of two wood mouse populations (Apodemus sylvaticus L.) were carried out outside the village Vík. S. Iceland, and in a birch forest close to the lake Mývatn, N. Iceland. The population at Vík was studied on a 1.10 ha grid on a slope covered with Angelica archangelica L., the seeds of which constituted a major food resource for the mice. Altogether 948 individuals were caught, marked and followed during 1973–1978. No other free-living small mammals occur in Iceland, and predator pressure is low and considered non-existent on the Angelica slope. The population density was extremely high; highest autumn density recorded was 150 ind ha^?1. Like other Apodemus populations, the one at Vík showed a typical seasonal pattern in numbers: a winter decrease, relatively stable numbers in late summer, and an autumn peak. Breeding started in late April or early May and ended in late September; no winter breeding was observed. Three or four litters were produced, and the last litter(s) of the season greatly contributed to the autumn increase. The output of young per reproductive female throughout the year was negatively correlated with spring density, and autumn peak density with the number of females in August. Also overwinter success and weight changes appeared to be associated with population density (and weather). Survival rates did not vary much between years and seasons and were consistently rather high. Autumn survival of juveniles was positively correlated with mean minimum temperatures. Juvenile survival was, at least at times, negatively affected by the presence of adult males. Trapping outside the main grid did not indicate any marked dispersal, though some immigration to the grid must have occurred, especially in spring and early summer. Among the factors governing the population density and dynamics of wood mouse at Vík we emphasize the absence of interspecific competition and predation, the rich and dependable food resources (mainly Angelica seeds), the mild (though moist and windy) climate, the shelter of dense Angelica stands, the abundance of crevices (e.g. nest-sites), and the social interactions. The birch forest in northern Iceland was low productive and the wood mouse population differed from the Vík population by exhibiting low density, short breeding season, and high vagility; characteristics we consider to be representative for most Apodemus populations in Iceland.