Rock2 promotes the invasion and metastasis of hepatocellular carcinoma by modifying MMP2 ubiquitination and degradation

Rho-associated coiled-coil-containing protein kinase 2 (Rock2) is a downstream effector of Rho that plays an important role in the tumorigenesis and progression of hepatocellular carcinoma (HCC). Matrix metalloproteinase 2 (MMP2) is a master regulator of tumor metastasis. In this study, we investigated the collections of Rock2 and MMP2 in HCCs and determined the potential role and molecular mechanism of Rock2 in MMP2-mediated invasiveness and metastasis. We found that Rock2 and MMP2 were markedly overexpressed in HCCs compared with the corresponding adjacent tissues, where a positive correlation in their expression was found. The knockdown of Rock2 significantly decreased MMP2 expression and inhibited the invasion and metastasis of HCC in vitro and in vivo. Additionally, the upregulation of MMP2 rescued the decreased migration and invasion induced by the knockdown of Rock2, whereas the knockdown of MMP2 decreased Rock2-enhanced HCC migration and invasion. Mechanistically, Rock2 stabilized MMP2 by preventing its ubiquitination and degradation. Together, our results link two drivers of invasion and metastasis in HCC and identify a novel pathway for MMP2 control.     

Patines et cortex de météorisation de grès du site rupestre d’Oum La Leg (Anti-Atlas, Maroc)

Within the framework of the European Commission's program “Patine du Desert” (DG Research; INCO-CT-FP6-2004-509100), partially concerning the preservation of Saharan engraved rock art, the approach led to the study of the sandstone patina from the rock art site of Oum La Leg (Anti-Atlas, Morocco). The rock and its Tazina school engravings are sometimes covered with a thin brown-ochre coating which only shows a dark patina, the desert patina. This film corresponds to a weathering cortex whose formation is contemporary with wet events from the Holocene period. On the surface, the patinas’ diversity is linked to the heterogeneous distribution of manganese oxides (birnessite and todorokite) due to a reorganisation which led to the incorporation of aeolian silts. Some dating suggestions are made to establish an ante quem age for the engraved lines.     

Context‐specific influence of water temperature on brook trout growth rates in the field

1. Modelling the effects of climate change on freshwater fishes requires robust field‐based estimates accounting for interactions among multiple factors. 2. We used data from an 8‐year individual‐based study of a wild brook trout (Salvelinus fontinalis) population to test the influence of water temperature on season‐specific growth in the context of variation in other environmental (i.e. season, stream flow) or biotic factors (local brook trout biomass density and fish age and size) in West Brook, a third‐order stream in western Massachusetts, U.S.A. 3. Changes in ambient temperature influenced individual growth rates. In general, higher temperatures were associated with higher growth rates in winter and spring and lower growth rates in summer and autumn. However, the effect of temperature on growth was strongly context‐dependent, differing in both magnitude and direction as a function of season, stream flow and fish biomass density. 4. We found that stream flow and temperature had strong and complex interactive effects on trout growth. At the coldest temperatures (in winter), high stream flows were associated with reduced trout growth rates. During spring and autumn and in typical summers (when water temperatures were close to growth optima), higher flows were associated with increased growth rates. In addition, the effect of flow at a given temperature (the flow‐temperature interaction) differed among seasons. 5. Trout density negatively affected growth rate and had strong interactions with temperature in two of four seasons (i.e. spring and summer) with greater negative effects at high temperatures. 6. Our study provided robust, integrative field‐based estimates of the effects of temperature on growth rates for a species which serves as a model organism for cold‐water adapted ectotherms facing the consequences of environmental change. Results of the study strongly suggest that failure to derive season‐specific estimates, or to explicitly consider interactions with flow regime and fish density, will seriously compromise our ability to predict the effects of climate change on stream fish growth rates. Further, the concordance we found between empirical observations and likely energetic mechanisms suggests that our general results should be relevant at broader spatial and temporal scales.     

Effects of coalbed natural gas development on fish assemblages in tributary streams of the Powder and Tongue rivers

1. Extraction of coalbed natural gas (CBNG) often results in disposal of large quantities of CBNG product water, which may affect aquatic ecosystems. We evaluated the effects of CBNG development on fish assemblages in tributary streams of the Powder and Tongue rivers. We used treatment and control, impact versus reference sites comparisons, surveys of CBNG product‐water streams and in situ fish survival approaches to determine if CBNG development affected fish assemblages. 2. Several of our results suggested that CBNG development did not affect fish assemblages. Species richness and index of biotic integrity (IBI) scores were similar in streams with and streams without CBNG development, and overall biotic integrity was not related to the number or density of CBNG wells. Fish occurred in one stream that was composed largely or entirely of CBNG product water. Sentinel fish survived in cages at treatment sites where no or few fish were captured, suggesting that factors such as lack of stream connectivity rather than water quality limited fish abundance at these sites. Fish species richness did not differ significantly from 1994 to 2006 in comparisons of CBNG‐developed and undeveloped streams. Biotic integrity declined from 1994 to 2006; however, declines occurred at both impact and reference sites, possibly because of long‐term drought. 3. Some evidence suggested that CBNG development negatively affected fish assemblages, or may do so over time. Specific conductivity was on average higher in treatment streams and was negatively related to biotic integrity. Four IBI species richness metrics were negatively correlated with the number or density of CBNG wells in the catchment above sampling sites. Bicarbonate, one of the primary ions in product water, was significantly higher in developed streams and may have limited abundance of longnose dace (Rhinichthys cataractae). Total dissolved solids, alkalinity, magnesium and sulphate were significantly higher in developed streams. 4. Biological monitoring conducted before the development of CBNG, and continuing through the life of development and reclamation, together with data on the quantity, quality and fate of CBNG product water will allow robust assessment of potential effects of future CBNG development worldwide.     

Leafminers help us understand leaf hydraulic design

Leaf hydraulics of Aesculus hippocastanum L. were measured over the growing season and during extensive leaf mining by the larvae of an invasive moth (Cameraria ohridella Deschka et Dimic) that specifically destroy the palisade tissue. Leaves showed seasonal changes in hydraulic resistance (R_lamina) which were related to ontogeny. After leaf expansion was complete, the hydraulic resistance of leaves and the partitioning of resistances between vascular and extra‐vascular compartments remained unchanged despite extensive disruption of the palisade by leafminers (up to 50%). This finding suggests that water flow from the petiole to the evaporation sites might not directly involve the palisade cells. The analysis of the temperature dependence of R_lamina in terms of Q₁₀ revealed that at least one transmembrane step was involved in water transport outside the leaf vasculature. Anatomical analysis suggested that this symplastic step may be located at the bundle sheath where the apoplast is interrupted by hydrophobic thickening of cell walls. Our findings offer some support to the view of a compartmentalization of leaves into well‐organized water pools so that the transpiration stream would involve veins, bundle sheath and spongy parenchyma, while the palisade tissue would be largely by‐passed with the possible advantage of protecting cells from short‐term fluctuations in water status.     

A PCR-based method for diet analysis in freshwater organisms using 18S rDNA barcoding on faeces

The development of DNA barcoding from faeces represents a promising method for animal diet analysis. However, current studies mainly rely on prior knowledge of prey diversity for a specific predator rather than on a range of its potential prey species. Considering that the feeding behaviour of teleosts may evolve with their environment, it could prove difficult to establish an exhaustive listing of their prey. In this article, we extend the DNA barcoding approach to diet analysis to allow the inclusion of a wide taxonomic range of potential prey items. Thirty-four ecological clade-specific primer sets were designed to cover a large proportion of prey species found in European river ecosystems. Selected primers sets were tested on isolated animal, algal or plant tissues and thereafter on fish faeces using nested PCR to increase DNA detection sensitivity. The PCR products were sequenced and analysed to confirm the identity of the taxa and to validate the method. The methodology developed here was applied to a diet analysis of three freshwater cyprinid species that are assumed to have similar feeding behaviour [Chondrostoma toxostoma toxostoma (Vallot 1837), Chondrostoma nasus nasus (Linnaeus, 1758) and Barbus barbus, (Linneaus 1758)]. These three species were sampled in four different hydrographic basins. Principal Component Analysis based on prey proportions identified distinct perilithon grazer and benthophagous behaviours. Furthermore, our results were consistent with the available literature on feeding behaviour in these fish. The simplicity of the PCR-based method and its potential generalization to other freshwater organisms may open new perspectives in food web ecology.     

Inter‐regional comparison of land‐use effects on stream metabolism

1. Rates of whole‐system metabolism (production and respiration) are fundamental indicators of ecosystem structure and function. Although first‐order, proximal controls are well understood, assessments of the interactions between proximal controls and distal controls, such as land use and geographic region, are lacking. Thus, the influence of land use on stream metabolism across geographic regions is unknown. Further, there is limited understanding of how land use may alter variability in ecosystem metabolism across regions. 2. Stream metabolism was measured in nine streams in each of eight regions (n = 72) across the United States and Puerto Rico. In each region, three streams were selected from a range of three land uses: agriculturally influenced, urban‐influenced, and reference streams. Stream metabolism was estimated from diel changes in dissolved oxygen concentrations in each stream reach with correction for reaeration and groundwater input. 3. Gross primary production (GPP) was highest in regions with little riparian vegetation (sagebrush steppe in Wyoming, desert shrub in Arizona/New Mexico) and lowest in forested regions (North Carolina, Oregon). In contrast, ecosystem respiration (ER) varied both within and among regions. Reference streams had significantly lower rates of GPP than urban or agriculturally influenced streams. 4. GPP was positively correlated with photosynthetically active radiation and autotrophic biomass. Multiple regression models compared using Akaike’s information criterion (AIC) indicated GPP increased with water column ammonium and the fraction of the catchment in urban and reference land‐use categories. Multiple regression models also identified velocity, temperature, nitrate, ammonium, dissolved organic carbon, GPP, coarse benthic organic matter, fine benthic organic matter and the fraction of all land‐use categories in the catchment as regulators of ER. 5. Structural equation modelling indicated significant distal as well as proximal control pathways including a direct effect of land‐use on GPP as well as SRP, DIN, and PAR effects on GPP; GPP effects on autotrophic biomass, organic matter, and ER; and organic matter effects on ER. 6. Overall, consideration of the data separated by land‐use categories showed reduced inter‐regional variability in rates of metabolism, indicating that the influence of agricultural and urban land use can obscure regional differences in stream metabolism.     

The Potential Role of Stream Habitat Restoration in Facilitating Salmonid Invasions: A Habitat‐Hydraulic Modeling Approach

Many successful invasions have taken place in systems where harmful disturbance has changed habitat conditions. However, less attention has been paid to the role of habitat restoration, which modifies habitats and thus also has the potential to facilitate invasions. We examined whether in‐stream habitat restorations have the potential to either facilitate or resist invasion by two widely introduced non‐native stream salmonids, Salvelinus fontinalis Mitchill and Oncorhynchus mykiss Walbaum, in Finland. A physical habitat simulation system was used to calculate whether the habitat area for the target species increased or decreased following the restorations. For comparison, we also reported results for four native stream fish species. The simulations showed that the restored streams provided the highest amount of usable habitat area for the native species, particularly for Salmo salar L. and Gottus gobio L. However, it was interesting to note that the restorations significantly increased habitat quality for the two non‐native species, especially at low flows. Nevertheless, the non‐native species had the lowest amount of usable habitat area overall. The modeling results indicated that not only habitat destruction but also habitat restoration could contribute to the spread of non‐native species. Fisheries and wildlife managers should be aware of the possibility, when restoring habitats in order to preserve native ecosystems, that non‐native species could manage to gain a foothold in restored habitats and use them as population sources for further spread. Knowing the widespread negative effect of non‐native species, this risk should not be underestimated.     

Rapid morphological change in stream beetle museum specimens correlates with climate change

Abstract 1. Climate change has been occurring at unprecedented rates and its impacts on biological populations is beginning to be well documented in the literature. For many species, however, long‐term records are not available, and trends have not been documented. 2. Using museum specimens from southern USA, we show that the stream‐dwelling beetle Gyretes sinuatus has shown an 8% increase in body size and change in body shape (fineness ratio) from 1928 to 1988. Any directional morphological change observed over time could be an indicator of a microevolutionary response. 3. During these 60 years, there have also been changes in temperature, precipitation, and location of collection sites. Unlike the global trend, mean annual temperature in the region has decreased, and furthermore, total annual precipitation has increased. By investigating how these various ecological and geographical variables may affect body size and shape, we can examine which pressures may promote larger and/or thinner beetles. 4. Results indicate that mean annual temperature was the most predictive variable for the change in size and shape. We suggest there is an adaptive role for temperature on body size and shape of stream dwelling organisms. 5. We found that museum specimens can be invaluable resources of information when collection date and location information is available. We promote the use of such specimens for future studies of the morphological response to climate change.