EXTRA-LYSOSOMAL PROTEOLYSIS AND EXPRESSION OF CALPAINS AND CALPASTATIN IN CULTURED THYROID CELLS

Proteolysis at neutral pH in the soluble fraction of cultured pig thyroid epithelial cells was examined using a synthetic calpain substrate, succinyl-Leu-Tyr-7-amino-4-methylcoumarin. The Ca²⁺-independent proteolytic activity was largely inhibited by substances known to affect cysteine- and metalloproteases, whereas no or little effects were obtained with inhibitors affecting serine- and aspartic proteases. Addition of Ca²⁺did not significantly alter the rate of substrate degradation. Biochemical separation via hydrophobic interaction chomatography and Western blotting demonstrated the presence of both m-calpain (40% of total calpain) and μ-calpain (60%) in confluent thyrocytes. Determination of calpastatin activity indicated a 30 times higher level of the inhibitor as compared to total calpain activity. Western blotting showed the presence of a 110kD calpastatin form with additional low mol wt forms possibly representing fragmentation products. In immunofluorescent stainings, m-calpain had a diffuse cytoplasmic distribution whereas μ-calpain was located both in the cytoplasm and at the cell—cell contacts. Calpastatin immunoreactivity was mainly granular and located close to the nucleus, although a fibrillar distribution was also observed. The results show the presence of all components of the calpain/calpastatin system and indicate a strict control of calpain activity in cultured thyrocytes. The different subcellular distributions of calpains and calpastatin suggests that they are compartmentalized and require mobilization to interact.     

Hydrologic effects on riparian vegetation in a boreal river: an experiment testing climate change predictions

Climate change is expected to alter the magnitude and variation of flow in streams and rivers, hence providing new conditions for riverine communities. We evaluated plant ecological responses to climate change by transplanting turfs of riparian vegetation to new elevations in the riparian zone, thus simulating expected changes in water‐level variation, and monitored the results over 6 years. Turfs moved to higher elevations decreased in biomass and increased in species richness, whereas turfs transplanted to lower elevations gained biomass but lost species. Transplanted plant communities responded slowly to the new hydrologic conditions. After 6 years, biomass of transplanted turfs was statistically indistinguishable from target level controls, but species richness and species composition of transplants were intermediate between original and target levels. By using projections of future stream flow according to IPCC climate change scenarios, we predict likely changes to riparian vegetation in boreal rivers. Climate‐driven hydrologic changes are predicted to result in narrower riparian zones along the studied Vindel River in northern Sweden towards the end of the 21st century. Present riparian plant communities are projected to be replaced by terrestrial communities at high elevations as a result of lower‐magnitude spring floods, and by amphibious or aquatic communities at low elevations as a result of higher autumn and winter flows. Changes to riparian vegetation may be larger in other boreal climate regions: snow melt fed spring floods are predicted to disappear in southern parts of the boreal zone, which would result in considerable loss of riparian habitat. Our study emphasizes the importance of long‐term ecological field experiments given that plant communities often respond slowly and in a nonlinear fashion to external pressures.     

Facilitation and interference among three predators affect their consumption of a stream-dwelling mayfly

1. We experimentally tested if a multiplicative risk model accurately predicted the consumption of a common mayfly at risk of predation from three predator species in New Zealand streams. Deviations between model predictions and experimental observations were interpreted as indicators of ecologically important interactions between predators. 2. The predators included a drift‐feeding fish [brown trout (T), Salmo trutta], a benthivorous fish [galaxiid (G), koaro, Galaxias brevipennis] and a benthic predatory stonefly (S; Stenoperla sp.) with Deleatidium sp. mayflies as prey. Eight treatments with all predator species combinations and a predator‐free control were used. Experiments were performed in aquaria with cobbles as predator refuges for mayflies and we measured the proportion of prey consumed after 6 h for both day and night trials. 3. Trout consumed a higher proportion of prey than other predators. For the two predator treatments we found less than expected prey consumption in the galaxiid + trout treatment (G + T) for both day and night trials, whereas a higher than expected proportion of prey was consumed during night time in the stonefly + trout (S + T) treatment. 4. The results indicate interference (G + T) and facilitation (S + T) between predators depending on predator identity and time of day. Thus, to make accurate predictions of interspecific interactions, it is necessary to consider the ecology of individual species and how differences influence the direction and magnitude of interactions.     

Floral scents in butterfly-pollinated plants: possible convergence in chemical composition

The study explores whether or not there are convergent patterns in floral scent composition among plant species that completely or partially rely on butterflies for pollination. Floral scent compounds were analysed from 22 flowering butterfly-pollinated plant species, representing 13 families which originate mainly from temperate North Europe but also from tropical and temperate America. Scents were collected using the dynamic headspace adsorption method and identified with coupled gas chromatography and mass spectrometry (GC-MS). In total, 217 floral scent compounds were identified, with the number per species ranging from 8 to 65. The major emerging pattern is the occurrence of certain compounds emitted exclusively by the flowers of many of the investigated species in major amounts – the benzenoids phenylacetaldehyde and 2-phenylethanol, the monoterpenes linalool and linalool oxide (furanoid) I and II and the irregular terpene oxoisophorone. It is likely that these compounds serve as a signal to attract pollinating butterflies, and may have evolved in conjunction with the sensory capabilities of butterflies as a specific group of pollinators. While there is convergence in terms of the compounds sharing this function there has been a geographical divergence in terms of their relative abundance. The predominance (in terms of both numbers and relative amount) of benzenoids in many of the scent blends of the European temperate species and of linalool and its derivatives in those of the American species constitute two discernible groups among these plants.  © 2002. The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 140 , 129–153.     

Static and dynamic bending has minor effects on xylem hydraulics of conifer branches (Picea abies,Pinus sylvestris)

The xylem hydraulic efficiency and safety is usually measured on mechanically unstressed samples, although trees may be exposed to combined hydraulic and mechanical stress in the field. We analysed changes in hydraulic conductivity and vulnerability to drought‐induced embolism during static bending of Picea abies and Pinus sylvestris branches as well as the effect of dynamic bending on the vulnerability. We hypothesized this mechanical stress to substantially impair xylem hydraulics. Intense static bending caused an only small decrease in hydraulic conductance (?19.5 ± 2.4% in P. abies) but no shift in vulnerability thresholds. Dynamic bending caused a 0.4 and 0.8 MPa decrease of the water potential at 50 and 88% loss of conductivity in P. sylvestris, but did not affect vulnerability thresholds in P. abies. With respect to applied extreme bending radii, effects on plant hydraulics were surprisingly small and are thus probably of minor eco‐physiological importance. More importantly, results indicate that available xylem hydraulic analyses (of conifers) sufficiently reflect plant hydraulics under field conditions.     

Evaluating stocking of YOY pike Esox lucius as a tool in the restoration of shallow lakes

1. Stocking of piscivores in shallow, eutrophicated lakes to reduce cyprinid densities is a common approach in lake restorations. Young‐of‐the‐year (YOY) pike Esox lucius are frequently used to reduce cyprinid densities, but their effectiveness is equivocal. This study uses a simple model to assess the efficacy of pike stocking as a lake restoration tool. 2. The model evaluates YOY pike consumption in relation to different survival and growth patterns, timing of stocking, degree of piscivory of pike as well as YOY cyprinid growth patterns and productivity. The parameters used in the model are obtained from field surveys in eight study lakes stocked with YOY pike as well as from the literature. 3. Our model showed that all parameters studied were important for predicting the effects of pike stocking on cyprinids. In particular, body size at stocking, cyprinid production and pike survival were good predictors of biomanipulation success. 4. However, our model showed that only under very specific circumstances will stocking of YOY pike result in a reduction of YOY cyprinid densities large enough to affect water quality. 5. The findings of this study question the efficacy of pike stocking as an appropriate and reliable tool for restoration programmes in shallow lakes, and indicate that when used efforts should be made to optimize the timing of stocking in relation to YOY cyprinid production and to increase the stocking body size of the pike.