Cell division in the unicellular microalga Dunaliella viridis depends on phosphorylation of extracellular signal-regulated kinases (ERKs)

In mammalian cells, MAPKs are involved in both stress response (JNK and p38 pathways) and cell proliferation and differentiation [extracellular signal-regulated kinase (ERK)] through protein kinase cascades. Exposure of Dunaliella viridis cell cultures to PD98059, a very specific inhibitor of the ERK signalling pathway, resulted in a total arrest of cell proliferation and a complete dephosphorylation of ERK. As shown by flow cytometry analysis of propidium iodide-stained cells, PD98059 stopped mitosis at the G(2) phase after the S phase has been completed. Multiple physiological parameters such as cell motility and reducing power generation (NADPH) clearly indicate that the treated cells are wholly viable. Exposure of D. viridis to environmental stresses that impair cell division, such as hyperosmotic shock, nitrogen starvation, or sublethal UV irradiation, caused a marked decrease in the phospho-ERK levels as detected by western blot. Two 400 bp polynucleotides from D. viridis with high homologies to published sequences of ERK1 and ERK2 were cloned, sequenced, and submitted to GenBank. Northern blot analysis revealed two mRNA bands of approximately 1.9 kb, consistent with the expected size of ERK proteins ( approximately 40 kDa). Sequence analysis showed that they contained several mitogen-activated protein kinase (MAPK) conserved domains, including II, III, VIb, VII, and the double phosphorylation motif. Interestingly, in D. viridis, this motif was T*DY* instead of the canonic T*EY*. Based on this finding, ERK plant sequences can be divided into two groups, one termed the T*DY* branch and the other termed the T*EY* branch. The molecular and functional data presented here suggest that ERK is a very ancient signalling pathway and that it was already present in the last common ancestor of all eukaryotic cells.     

Expression of Chlorovirus MT325 aquaglyceroporin (aqpv1) in tobacco and its role in mitigating drought stress

Main conclusions

A Chlorovirus aquaglyceroporin expressed in tobacco is localized to the plastid and plasma membranes. Transgenic events display improved response to water deficit. Necrosis in adult stage plants is observed. Aquaglyceroporins are a subclass of the water channel aquaporin proteins (AQPs) that transport glycerol along with other small molecules transcellular in addition to water. In the studies communicated herein, we analyzed the expression of the aquaglyceroporin gene designated, aqpv1, from Chlorovirus MT325, in tobacco (Nicotiana tabacum), along with phenotypic changes induced by aqpv1 expression in planta. Interestingly, aqpv1 expression under control of either a constitutive or a root-preferred promoter, triggered local lesion formation in older leaves, which progressed significantly after induction of flowering. Fusion of aqpv1 with GFP suggests that the protein localized to the plasmalemma, and potentially with plastid and endoplasmic reticulum membranes. Physiological characterizations of transgenic plants during juvenile stage growth were monitored for potential mitigation to water dry-down (i.e., drought) and recovery. Phenotypic analyses on drought mimic/recovery of juvenile transgenic plants that expressed a functional aqpv1 transgene had higher photosynthetic rates, stomatal conductance, and water use efficiency, along with maximum carboxylation and electron transport rates when compared to control plants. These physiological attributes permitted the juvenile aqpv1 transgenic plants to perform better under drought-mimicked conditions and hastened recovery following re-watering. This drought mitigation effect is linked to the ability of the transgenic plants to maintain cell turgor.     

Community structure of fish in lowland streams differ substantially between subtropical and temperate climates

Fish are important in the structuring of other communities and may have large effects on the functioning of aquatic ecosystems. The structure of fish communities, in turn, seems to differ with climate. We compared the characteristics of fish assemblages in lowland streams located in two contrasting climates (cold-temperate Europe and subtropical South America) by use of published and unpublished data on streams of similar depth, width, and slope (n total?=?91 streams). We also selected a subset of seven comparable little-affected streams in the two contrasting climates: temperate (Denmark, 55°?C57°N, Dk) and subtropical (Uruguay, 30°?C35°S, Uy) and compared the fish community structures in relation to environmental characteristics. We then analysed a series of potential explanatory factors behind the patterns observed, in particular the effect of ambient temperature, by comparing temperature-corrected community metabolism. Significantly higher species richness, higher densities, lower biomass, smaller mean body size, and lower mean weight of fish were observed for the subtropical streams than for the temperate streams, both in the literature review and in the subset of streams. Several characteristics of fish assemblages in streams may be explained by direct and indirect effects of temperature. Accordingly, fish in subtropical systems had a temperature-corrected community metabolism I?m^?2 equal to that of fish in temperate systems, indicating that temperature, besides historical factors, is an important driver of different size structures. Our findings concur with differences previously found in littoral areas of shallow lakes, suggesting that these patterns are not restricted to running waters. Our results elucidate how fish community structure might be affected by increases in temperature triggered by climate warming.     

Session-to-session variations in external load measures during small-sided games in professional soccer players

The aims of this study were 1) to analyse session-to-session variations in different external load measures and 2) to examine differences in within-session intervals across different small-sided game (SSG) formats in professional players. Twenty professional soccer players (mean ± SD; age 28.1 ± 4.6 years, height 176.7 ± 4.9 cm, body mass 72.0 ± 7.8 kg, and body fat 10.3 ± 3.8%) participated in 3v3, 4v4, and 6v6 SSGs under different conditions (i.e., touch limitations and presence of goalkeepers vs. free touch and ball possession drill) over three sessions. Selected external load measures—including total distance (TD), high-intensity running (HIR, distance covered > 14.4 km^.h^-1), high-speed running (HSR, distance covered > 19.8 km^.h^-1), and mechanical work (MW, accelerations and deceleration > 2.2 m^.s²)—were recorded using GPS technology during all SSG sessions. Small to large standardized typical errors were observed in session-to-session variations of selected measures across SSGs. TD^.min^-1 showed less variability, having a coefficient of variation (CV) of 2.2 to 4.6%, while all other selected external load measures had CV values ranging from 7.2% to 29.4%. Trivial differences were observed between intervals in TD^.min^-1 and HIR^.min^-1 for all SSGs, as well as in HSR^.min^-1 and MW^.min^-1 for most SSG formats. No reductions or incremental trends in session-to-session variations were observed when employing touch limitations or adding goalkeepers. The increased noise observed in higher speed zones (e.g., high-speed running) suggests a need for more controlled, running-based conditional drills if the aim is greater consistency in these measures.     

A systematic review of collective tactical behaviour in futsal using positional data

Although many studies on collective tactical behaviour have been published in the last decade, no study has revised and summarized the findings provided for futsal. The main aim of this systematic review was to identify and discuss the geometrical centre (GC), distance and area tactical variables used to assess team behaviour in futsal. In addition, it summarizes the findings on the tactical response during futsal competition and training. A systematic review of the relevant articles provided on futsal was carried out using seven electronic databases (SPORTDiscus, ProQuest, Cochrane Plus, Scopus, Google Scholar, PubMed and Web of Science) until September 25, 2019. From a total of 1,209 studies initially found, 12 were included in the qualitative synthesis. There were some trends in the analysis of positional data in futsal with the most relevant situations analysed being 1 vs 1 and 5 vs 4+Goalkeeper. The distances and angles between two points were the most assessed tactical variables. Five types of distance variables were used to assess collective tactical behaviour in futsal: GC-GC, GC-player, player-player, player-ball and player-space. Pressure (GC-GC) was greater in shots on goal than in tackles during professional futsal matches. Area variables were reduced to occupied space, exploration space and dominant area. Occupied space was measured only during competition while the dominant area was measured only during training sessions. The surface area and dominant regions were greater when players were attacking in comparison to when they were defending. In addition, two non-linear techniques (i.e. relative phase and entropy) were applied to analyse synchronisation and complexity and regularity or predictability. Defenders were highly synchronous, while attackers tried to break this coordination to achieve possibilities for action. Task constraints are suitable to induce different regularity patterns. This review is an opportunity to develop studies aimed at bridging the gap in collective tactical behaviour in futsal.     

Conicamin, a novel histamine antagonist from the mediterranean tunicate Aplidium conicum

In addition to the known 6-bromo-hypaphorine (2) and plakohypaphorine-A (3), the methanol extract of the Mediterranean tunicate Aplidium conicum was shown to contain conicamin, a novel indole alkaloid having histamine-antagonistic activity which structure was determined to be 1 on the basis of the spectral data.