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Aims: The restoration of unused urban‐industrial areas has great conservation potential as it can sometimes create a win‐win situation in terms of nature conservation and economic benefits. Financial data of restoration projects are rarely published. To close this gap, we report the cost–benefit analysis of different landscaping methods used in the greening of the outskirts of a factory industrial area. Location: The study area is located in a factory area in Nyíregyháza, Nyírség, Hungary. Methods: In this study, we compare the costs of the implementation and maintenance of three different types of grasslands (intensively and extensively managed park lawns and semi‐natural grassland, called eco area) and the ecological benefits in terms of vegetation and potential for pollinators. Results: The implementation and maintenance costs of the extensively managed park lawn were more than ten‐fold higher than that of the eco area, despite the higher price of native plant material. The construction of the extensive park lawn is the cheapest, but the eco area pays off in ca. 1.5 years compared to it, taking into account the lower maintenance costs. Seven years after implementation, the eco area is the most species‐rich, hosting the highest number of natural constituent species, but with similar vegetation cover as in the intensively managed park lawn without costly maintenance. The cover and species richness of legumes and forbs are also higher in the eco area, ensuring a food source for pollinators. Conclusions: Our results support that introducing species‐rich and diverse grasslands with native, habitat specialist species as an alternative to traditional landscaping in urban‐industrial areas can be economically and ecologically rewarding. The higher costs of planning, plant material, and implementation pay off in a short time due to low maintenance costs and can lead to a win‐win situation for nature as well as for the local economy. [ABSTRACT FROM AUTHOR]

Aichi Target 15 aims to enhance the resilience of ecosystems through the restoration of 15% of degraded land by 2020. Compliance with the target is still insufficient, partly due to the lack of appropriate baselines and knowledge of restoration efforts. The four-level model of ecosystem condition was suggested to set the baseline and support the estimation of progress. This study is the first to assess the condition of MAES ecosystem types at a country scale, using the suggested approach. Altogether 613,000 ha of land is in close to natural condition (Level 1), 893,000 ha in a slightly degraded state (Level 2), 1,907,000 ha in moderately degraded condition (Level 3), and 5,890,000 ha of land is in highly modified category (Level 4). An inventory of ecological restoration interventions was created based on the reports of the national park directorates and EU LIFE projects. The findings of the restoration database of 634 interventions were compared to the need for restoration of nine habitat types. Restoration efforts were focused at habitat types with the most extended degraded area, except for dry forest restoration and the fight against invasive species in all three forest types. This study provides a more reliable estimate of habitats in natural state than Habitat Directive reporting can achieve, and is therefore better suited for estimating compliance with the 15% target. Only 1% of degraded land was the target for restoration in the period of 2002–2016. The details of the survey regarding habitat and degradation types can support further restoration planning. [ABSTRACT FROM AUTHOR]

Scaling-up ecological restoration demands the involvement of private sector actors. Experience regarding science-based habitat restoration programs in the sector should be made available to support further joint projects. In our case, hierarchical restoration prioritization was applied to select best target for habitat reconstruction at a Hungarian industrial area. Multiple potential natural vegetation model, a novel approach, supported restoration prioritization satisfying both ecological (sustainability and nature conservation value) and other needs (feasibility, rapid green surface, amenity, and education value). The target that met all priorities was the open steppe forest that has a mosaic arrangement with open and closed sand steppes. The potential area of this xero-thermophile oak wood is expected to expand in Hungary with climate change, therefore the selected target has a likelihood of long-term sustainability, if established. A matrix of sand steppes was created first at the factory area in 2014-2015, and tree and shrub saplings were planted in this matrix. The seeding induced rapid changes in vegetation composition: the second year samples became close to reference sand steppes in the principal component analysis ordination space. Tree and shrub survival was species dependent, reaching a maximum of 52 and 73% for tree and shrub species, respectively. One tree and 2 shrub species did not survive at all. Altogether 53 of 107 target species have established. So far, restored vegetation development confirmed the suitability of the applied hierarchical prioritization framework at factory scale. [ABSTRACT FROM AUTHOR]

Successful use of woody species in reducing climatic and environmental risks of energy shortage and spreading pollution requires deeper understanding of the physiological functions controlling biomass productivity and phytoremediation efficiency. Targets in the breeding of energy willow include the size and the functionality of the root system. For the combination of polyploidy and heterosis, we have generated triploid hybrids (THs) of energy willow by crossing autotetraploid willow plants with leading cultivars (Tordis and Inger). These novel Salix genotypes (TH3/12, TH17/17, TH21/2) have provided a unique experimental material for characterization of Mid-Parent Heterosis (MPH) in various root traits. Using a root phenotyping platform, we detected heterosis (TH3/12: MPH 43.99%; TH21/2: MPH 26.93%) in the size of the root system in soil. Triploid heterosis was also recorded in the fresh root weights, but it was less pronounced (MPH%: 9.63–19.31). In agreement with root growth characteristics in soil, the TH3/12 hybrids showed considerable heterosis (MPH: 70.08%) under in vitro conditions. Confocal microscopy-based imaging and quantitative analysis of root parenchyma cells at the division–elongation transition zone showed increased average cell diameter as a sign of cellular heterosis in plants from TH17/17 and TH21/2 triploid lines. Analysis of the hormonal background revealed that the auxin level was seven times higher than the total cytokinin contents in root tips of parental Tordis plants. In triploid hybrids, the auxin–cytokinin ratios were considerably reduced in TH3/12 and TH17/17 roots. In particular, the contents of cytokinin precursor, such as isopentenyl adenosine monophosphate, were elevated in all three triploid hybrids. Heterosis was also recorded in the amounts of active gibberellin precursor, GA19, in roots of TH3/12 plants. The presented experimental findings highlight the physiological basics of triploid heterosis in energy willow roots. [ABSTRACT FROM AUTHOR]

The biomass productivity of the energy willow Salix viminalis as a short-rotation woody crop depends on organ structure and functions that are under the control of genome size. Colchicine treatment of axillary buds resulted in a set of autotetraploid S. viminalis var. Energo genotypes (polyploid Energo [PP-E]; 2n = 4x = 76) with variation in the green pixel-based shoot surface area. In cases where increased shoot biomass was observed, it was primarily derived from larger leaf size and wider stem diameter. Autotetraploidy slowed primary growth and increased shoot diameter (a parameter of secondary growth). The duplicated genome size enlarged bark and wood layers in twigs sampled in the field. The PP-E plants developed wider leaves with thicker midrib and enlarged palisade parenchyma cells. Autotetraploid leaves contained significantly increased amounts of active gibberellins, cytokinins, salicylic acid, and jasmonate compared with diploid individuals. Greater net photosynthetic CO2 uptake was detected in leaves of PP-E plants with increased chlorophyll and carotenoid contents. Improved photosynthetic functions in tetraploids were also shown by more efficient electron transport rates of photosystems I and II. Autotetraploidization increased the biomass of the root system of PP-E plants relative to diploids. Sections of tetraploid roots showed thickening with enlarged cortex cells. Elevated amounts of indole acetic acid, active cytokinins, active gibberellin, and salicylic acid were detected in the root tips of these plants. The presented variation in traits of tetraploid willow genotypes provides a basis to use autopolyploidization as a chromosome engineering technique to alter the organ development of energy plants in order to improve biomass productivity. [ABSTRACT FROM AUTHOR]

The role of complement in the regulation of T cell immunity has been highlighted recently by several groups. We were prompted to reinvestigate the role of complement receptor type 1 (CR1, CD3) in human T cells based on our earlier data showing that activated human T cells produce C3 (Torok et al. (2012) [48] ) and also by results demonstrating that engagement of Membrane Cofactor Protein (MCP, CD46) induces a switch of anti-CD3-activated helper T cells into regulatory T cells (Kemper et al. (2003) [17] ). We demonstrate here that co-ligation of CD46 and CD3, the two C3b-binding structures present on activated CD4+ human T cells significantly enhances CD25 expression, elevates granzyme B production and synergistically augments cell proliferation. The role of CR1 in the development of the Treg phenotype was further confirmed by demonstrating that its engagement enhances IL-10 production and reduces IFNγ release by the activated CD4+ T cells in the presence of excess IL-2. The functional in vivo relevance of our findings was highlighted by the immunohistochemical staining of tonsils, revealing the presence of CD4/CD3 double positive lymphocytes mainly in the inter-follicular regions where direct contact between CD4+ T cells and B lymphocytes occurs. Regarding the in vivo relevance of the complement-dependent generation of regulatory T cells in secondary lymphoid organs we propose a scenario shown in the figure. The depicted process involves the sequential binding of locally produced C3 fragments to CD46 and CD3 expressed on activated T cells, which – in the presence of excess IL-2 – leads to the development of Treg cells. [ABSTRACT FROM AUTHOR]

Soil nitrogen immobilization by carbon amendment is a management technique used for conservation purposes to increase the competitive ability of late-seral plant species over early-seral species based on their different tolerance of low soil nitrogen content., We immobilized nitrogen over six growing seasons on three ex-arable fields with poor sandy soils in the Hungarian lowland in order to restore endemic sandy grassland. Sucrose and sawdust were applied at rates based on previous laboratory experiments using local soils. We tested the efficacy of long-term carbon amendment for lowering soil nitrogen availability and favouring late-seral native species over early-seral weed species., Carbon amendments resulted in significant increase in soil microbial biomass C and reduced soil nitrogen availability after 2 years., Total vegetation cover was reduced by reducing soil nitrogen availability, but total species richness was not impacted. Cover of early-seral species decreased, and species richness and cover of late-seral species increased irrespective of nitrogen immobilization. However, after 4-6 years reducing soil nitrogen availability hampered the spread of moss under vascular vegetation., Synthesis and applications. This study supports the efficacy of carbon amendment as a tool to immobilize available soil nitrogen in the upper soil layers. However, the desired impact on vegetation was not fully achieved despite application over several years. Nitrogen immobilization was most relevant to bryophytes, lacking deep root systems, which may explain the responsiveness of this group to N limitation. The different impact of N availability on the complex of early-seral, late-seral vascular species and that of the bryophyte layer provides opportunity for directing state transformations in arid grasslands. Bryophyte cover can be suppressed through carbon amendments in order to enhance the germination and establishment of grassland species. The advantage of the method is that it opens bryophyte cover gradually without disturbing the soil surface, possibly avoiding the establishment of invasive species. However, further studies are required for deeper insight. [ABSTRACT FROM AUTHOR]

Abstract: Although the complement system is thought to be mainly involved in innate immunity and in the humoral arm of adaptive responses, evidence implicating that complement impacts T cell responses are accumulating recently. The role of the various activation products of the major complement component C3 were mainly studied so far in animal systems, and investigations regarding the effect of different C3-fragments on human T cells are sparse. Here we show that anti-CD3 activated human T lymphocytes derived from the blood and tonsil of healthy individuals produce C3, and the major cleavage fragment that appears on the T cell surface is iC3b. Based on studies carried out in allogenic system we demonstrate that the T cell membrane bound iC3b binds to the CR3 and probably to CR4 receptors expressed on monocyte-derived dendritic cells, and this interaction leads to significantly enhanced T-cell proliferation. Since neither C3aR and nor C3a binding could be detected on the membrane of anti-CD3 activated T cells, our findings indicate that in humans – in contrast to mice – the C3a peptide is most probably not involved directly in the T cell activation process. [Copyright &y& Elsevier]

Translocation from the cytosol to the nucleus is a major response by calmodulin (CAM) to stimulation of cells by Ca2+. However, the mechanisms involved in this process are still controversial and both passive and facilitated diffusion have been put forward. We tested nuclear translocation mechanisms in electroporated HeLa cells, rat cortical neurons and glial cells using novel calmodulin and inhibitor peptide probes and eonfocal microscopy. Passive diffusion of calmodulin across the nuclear membrane was measured in conditions in which facilitated transport was blocked and was compared to that of a similarly sized fluorescein-labeled dextran. Wheat germ agglutinin, which blocks facilitated transport but not passive diffusion, inhibited the nuclear entry of both wild-type and Ca2+-binding-deficient mutant calmodulin both in low and elevated [Ca2+]. Ca2+-dependent nuclear translocation was prevented by a membrane-permeant CaM inhibitor, the mTrp peptide, which indicated that it was specific to Ca2+/CaM. Diffusion of free CaM and Ca2+/CaM was considerably slower than the observed nuclear translocation by facilitated transport. Our data show that the majority of CaM nuclear entry occurred by facilitated mechanisms in all cell types examined, in part by a Ca2+-independent and in part by a Ca2+-dependent translocation mechanism. [ABSTRACT FROM AUTHOR]

The role of adenosine 5'-triphosphate (ATP) in the activation mechanism of α-Ca2+/calmodulin- dependent protein kinase II (αCaMKII) was investigated using the T286A non-autophosphorylatable mutant of αCaMKII. Characterization of the T286A-αCaMKII mutant revealed kcat = 0.06 ± 0.02 s-1 for the T286A mutant, a 6 (± 2)-fold lower value compared to wild-type αCaMKII with 100 µM smooth muscle myosin light chain (MLC) as substrate. MLC phosphorylation by the T286A mutant and wild-type αCaMKII was cooperative, with Hill coefficients 2.3 ± 0.1 and 2.4 ± 0.3, respectively. Km values for MLC were 96 ± 28 µM with T286A-αCaMKII and 49 ± 29 µM for wild-type αCaMKII. Thus, while the activity of αCaMKII was sensitive to mutation of the Thr286 residue to Ala, the mechanisms of the wild-type and T286A mutant enzyme appeared similar. Kd for Ca2+/calmodulin was 2-fold reduced to 40 nM compared to that of wild-type αCaMKII (75 nM). ATP induced a 9-fold stabilization of Ca2+/calmodulin binding to the T286A mutant enzyme. Fluorescence stopped-flow kinetic experiments revealed that two Ca2+/ calmodulin-enzyme complexes were formed, the first, unaffected by ATP, with association and dissociation rate constants of 2 × 107 M-1 s-1 and 5 s-1, respectively, containing calmodulin in extended conformation. The second complex, in which calmodulin adopted a compact conformation, was formed with association rate constant 3 × 106 M-1 s-1 and dissociation at 0.15 s-1 in the absence and 0.015 s-1 in the presence of ATP. These data show that ATP is involved in the activation mechanism by forming two classes of Ca2+/calmodulin·αCaMKII-ATP complex. It is likely that only one of the complexes is on the activation pathway. [ABSTRACT FROM AUTHOR]

Abstract The aim of the present study was to investigate the aggregation of cardiovascular risk factors (hyperinsulinaemia, impaired glucose tolerance, dyslipidaemia, and hypertension) in 180 (77 female, 103 male) obese and 239 control children. Blood glucose, serum insulin and lipid levels were determined from blood samples taken after an overnight fast. Oral glucose tolerance tests were performed and blood glucose concentrations were monitored. The body mass index, body fat (on the basis of skinfold measurements), lean body mass and waist/hip ratio were calculated and blood pressure was measured five times in all subjects. It was shown that only 14.4% of obese children were free from any risk factors, in contrast to 79.1% of the control children. Four risk factors (metabolic cardiovascular syndrome) were found in 8.9% of the obese children (8.7% in males and 9.1% in females) while none could be detected in controls. Considerable differences were also detected in the prevalence of one, two or three risk factors between control and obese children. Patients with the metabolic cardiovascular syndrome could not be characterized by any of the investigated anthropometric characteristics, but the duration of obesity was significantly longer in these children. Conclusion Potential risk factors for cardiovascular diseases already tend to cluster in childhood and they are strongly associated with obesity. Our observations suggest that the development of the metabolic cardiovascular syndrome has its origin in childhood. [ABSTRACT FROM AUTHOR]

Sarcoplasmic reticulum Ca2+ATPase is cleaved by trypsin at two sites, T1 and T2. Cleavage at T1 is complete, whereas only about 50% of the Ca2+ATPase is digested at the T2 site. In the absence of Ca2+ ionopbor, Ca2+- ATPase activity of the digested enzyme remains virtually unchanged. In the presence of Ca2+ ionophor, however, the calculated specific activity of the doubly cleaved Ca2+ATPase is decreased by about 40%. The decrease in Ca2+ transport activity is much more rapid than cleavage of the T2 site, and could he correlated with an increased leak of Ca2+ from the digested vesicles. We obtained evidence that this leakiness is independent of the digestion of the Ca2+ATPase itself and is presumably due to the digestion of some other components of the sarcoplasmic reticulum vesicles. Examination of steady-state phosphoenzyme levels resulting from phosphorylation by ATP and Pi, or dephosphorylation by ADP or ADP/EGTA revealed no difference between the digested and the undigested Ca2+ATPase indicating no change in the equilibria caused by the T2 cleavage. Analysis of the substrate concentration dependence of the Ca2+ ATPase activity also led to the conclusion that the digestion at T2 reduced the Vmax of ATP hydrolysis but leaves the Km unchanged. The above results are consistent with the model that cleavage at the T2 site reduces the turnover rate of the Ca2+ATPase reaction cycle by about 40% by slowing down or altering rite rate-limiting step without effecting the equilibrium constants of the examined steps. We found no evidence of true uncoupling of Ca2+ transport from ATP hydrolysis correlated with cleavage at the T2 site. [ABSTRACT FROM AUTHOR]

Objective To compare the outcomes of a 6-month-long accelerated rehabilitation with a 12-month-long rehabilitation. There is no consensus on the optimal duration of rehabilitation after anterior cruciate ligament reconstruction (ACLR). Trends in the past decades have shifted towards accelerated programs, often resulting in a return to play (RTP) at 4-6 months, postoperatively. However, longer rehabilitation cycles have recently experienced renaissance due to a greater understanding of graft remodeling. Methods Adult athletes who underwent ACLR between 2015 and 2018 by the same surgeon were included and followed-up prospectively for 24 months. Participants were allocated into two groups based on their RTP (6 months vs. 12 months) and compared with graft elongation, reoperation rate, and sports career (quit or continue) outcomes. Results Fifty-four patients underwent accelerated rehabilitation and 92 completed conventional rehabilitation. The accelerated rehabilitation was significantly associated with graft elongation--the accelerated rehabilitation group (n=9) and the conventional rehabilitation group (n=0), p<0.001--and need for reoperation--the accelerated rehabilitation group (n=5) and the conventional rehabilitation group (n=1), p=0.026. Although the relationship between rehabilitation time and quitting competitive sports did not reach significance at 0.05 level (p=0.063), it was significant when p<0.1, thereby showing a clear trend. Conclusion Accelerated rehabilitation increased graft elongation risk. Knee laxity ≥3 mm measured at 6 months after ACLR should be accompanied by RTP time frame re-evaluation. Arthrometry checkups or routine magnetic resonance imaging shortly after RTP may be considered in cases of accelerated rehabilitation. [ABSTRACT FROM AUTHOR]

The restoration of invasion-resistant plant communities is an important strategy to combat the negative impacts of alien invasions. Based on a systematic review and meta-analysis of seed-based ecological restoration experiments, here we demonstrate the potential of functional similarity, seeding density and priority effect in increasing invasion resistance. Our results indicate that native priority is the most promising mechanism to control invasion that can reduce the performance of invasive alien species by more than 50%. High-density seeding is effective in controlling invasive species, but threshold seeding rates may exist. Overall seeding functionally similar species do not have a significant effect. Generally, the impacts are more pronounced on perennial and grassy invaders and on the short-term. Our results suggest that biotic resistance can be best enhanced by the early introduction of native plant species during restoration. Seeding of a single species with high functional similarity to invasive alien species is unpromising, and instead, preference should be given to high-density multifunctional seed mixtures, possibly including native species favored by the priority effect. We highlight the need to integrate research across geographical regions, global invasive species and potential resistance mechanisms. A systematic review and meta-analyses of seed-based ecological restoration experiments identify native priority as a promising mechanism for controlling invasion of alien plant species, which can reduce the performance of invasive species by more than 50%. [ABSTRACT FROM AUTHOR]

Changes in the balance between glutamate (Glu) release and uptake may stimulate synaptic reorganization and even synapse loss. In the case of neurodegeneration, a mismatch between astroglial Glu uptake and presynaptic Glu release could be detected if both parameters were assessed independently and at a single-synapse level. This has now become possible due to a new imaging assay with the genetically encoded ultrafast Glu sensor iGluu. We report findings from individual corticostriatal synapses in acute slices prepared from mice of either sex that were > 1 year of age. Contrasting patterns of short-term plasticity and a size criterion identified two classes of terminals, presumably corresponding to the previously defined IT (intratelencephalic) and PT (pyramidal tract) synapses. The latter exhibited a higher degree of frequency potentiation/residual Glu accumulation and were selected for our first iGluu single-synapse study in Q175 mice, a model of Huntington’s disease (HD). In HD mice, the decay time constant of the perisynaptic Glu concentration (TauD), as an indicator of uptake, and the peak iG!u„ amplitude, as an indicator of release, were prolonged and reduced, respectively. Treatment of WT preparations with the astrocytic Glu uptake blocker TFB-TBOA (100 mu) mimicked the TauD changes in homozygotes. Considering the largest TauD values encountered in WT, —40% of PT synapses tested in Q175 heterozygotes can be classified as dysfunctional. Moreover, HD but not WT synapses exhibited a positive correlation between TauD and the peak amplitude of iGluu. Finally, EAAT2 (excitatory amino acid transport protein 2) immunoreactivity was reduced next to corticostriatal terminals. Thus, astrocytic Glu transport remains a promising target for therapeutic intervention. [ABSTRACT FROM AUTHOR]

Questions: The restoration of unused urban‐industrial areas has largely been neglected despite their great potential for nature conservation purposes. We applied three plant introduction treatments at a highly degraded industrial area to test whether plant material introduced onto industrial areas initiates plant assemblages similar in composition to those of reference grasslands. We specifically asked (a) Does restoration differ between primary and secondary reference sites in terms of overall species richness and cover three years after the introduction of plant material? (b) What is the sociability of species of the vegetation resulting from different treatments? Location: Restoration and reference grassland sites with acidic sandy soil in the temperate region of the EU, NE Hungary, Nyírség. Methods: As restoration techniques, we (a) directly seeded a single dominant species, (b) applied a commercial seed mixture, and (c) transferred hay. We compared species composition, cover, species richness and sociability of species in restoration treatments to reference grasslands. Inventories of vascular plant species were made at five randomly placed 2 m × 2 m sampling units per plot. Results: The species composition of seeded restoration plots (with a single dominant species and a commercial seed mixture) became similar to that of reference grasslands. Both types of seeding resulted in a similar cover, whereas seeding a commercial seed mixture and hay transfer resulted in a species richness similar to that of reference grasslands. The cover of the natural constituents (NC) of seeded plots also reached that of reference grasslands, while hay addition resulted in a lower cover and a higher number of NC than other methods. Conclusions: The introduction of propagules in degraded industrial areas can provide communities similar, after three years, to those of secondary reference grasslands that are 30 years of age. We conclude that investing in the restoration of native grasslands at unused urban‐industrial sites can be a great opportunity to enhance biodiversity. We applied three plant introduction treatments to restore sandy grasslands at an industrial area. The restored grassland was compared to reference grasslands. The treatments resulted in communities that, after three years, were similar to those of secondary reference grasslands with an age of 30 years. The restoration of native grasslands at unused urban‐industrial sites can therefore be a great opportunity to enhance biodiversity. [ABSTRACT FROM AUTHOR]

Question: What is the effect of invasive common milkweed (Asclepias syriaca L.) on the germination and early establishment of native grass species during open sand grassland vegetation recovery in old‐fields? Location: Fülöpháza Sand Dune Area, Hungary. Methods: A small‐scale experiment was carried out in a sandy old‐field infested with Asclepias. We designated 36 2 m × 2 m plots in patches of Asclepias. We seeded two native grass species Festuca vaginata and Stipa borysthenica in 12 plots each (a third of the plots was left unseeded). We applied repeated mechanical removal of Asclepias shoots on half of the plots for two growing seasons. The number and above‐ground cover of the two grass seedlings were evaluated for two growing seasons. Results: The number and above‐ground cover of Festuca and Stipa seedlings did not increase after applying Asclepias shoot removal during the 2 years of the study. We found lower seedling number and cover of Festuca in plots with Asclepias shoot removal in the second year, when a severe summer drought occurred at the study site. The number and cover of the Stipa seedlings did not differ between plots with Asclepias shoot removal and control plots throughout the experiment. Conclusions: We did not find any negative effects of the presence of the invasive Asclepias during open sand grassland regeneration in terms of germination and early establishment of the dominant grass species. We even detected a nurse effect of Asclepias on Festuca, where the shade of Asclepias may have mitigated the unfavourable abiotic conditions for Festuca caused by summer drought. This mitigation was not observed in the case of Stipa, which can better tolerate summer droughts. Our results suggest that Asclepias control is not required for a successful open sand grassland restoration in the early phase of vegetation recovery, and restoration efforts should focus on the mitigation of propagule limitation of native grasses. However, further information is needed about the effects of Asclepias on other elements of the biota and in later phases of secondary succession. We experimentally investigated the effect of the invasive herb common milkweed (Asclepias syriaca L.) on grassland recovery in terms of the establishment of two dominant grasses of the open sand grassland community. We observed that milkweed did not have any adverse effects on the early establishment of these species, and even had a nurse effect on one of them during summer drought. [ABSTRACT FROM AUTHOR]

Invasion of alien species is one of the main drivers of land degradation threatening both natural and managed ecosystems. Ecological restoration is crucial in controlling invasion to improve biotic resistance and avoid further land degradation. We investigated the possibility of controlling the establishment of invasive alien species (IAS) by native seed addition. We tested if trait similarity or increased propagule pressure of native species results in the suppression of IAS at the early stage of development. We set up a sowing experiment with three widespread IAS in Hungary of different life forms and functional groups (Asclepias syriaca, Conyza canadensis, Tragus racemosus) and four Pannonic sand grassland species (Festuca vaginata, Galium verum, Gypsophila paniculata, Saponaria officinalis). We found no significant differences in germination ability and seedling emergence between native species and IAS, despite the differences in thousand‐seed weight. Using univariate general linear models, we found that the seedling establishment of IAS can be reduced by adding native species at high densities but also depending on the species identity. Instead of species of similar traits, the seeding of a competitor perennial grass of sand grasslands (F. vaginata) reduced the seedling emergence of all studied IAS the most. Our results confirm that IAS can be effectively controlled by native seed addition in the early establishment stage, especially applying higher densities and competitive species. We conclude that invasion‐resistant restoration can be achieved by the combination of several factors, including high‐density sowing of native species that match IAS in the early stage of development. [ABSTRACT FROM AUTHOR]

Ca2+/calmodulin (Ca2+/CaM) interaction with connexins (Cx) is well-established; however, the mechanistic basis of regulation of gap junction function by Ca2+/CaM is not fully understood. Ca2+/CaM is predicted to bind to a domain in the C-terminal portion of the intracellular loop (CL2) in the vast majority of Cx isoforms and for a number of Cx-s this prediction has proved correct. In this study, we investigate and characterise both Ca2+/CaM and apo-CaM binding to selected representatives of each of the α, β and γ connexin family to develop a better mechanistic understanding of CaM effects on gap junction function. The affinity and kinetics Ca2+/CaM and apo-CaM interactions of CL2 peptides of β-Cx32, γ-Cx35, α-Cx43, α-Cx45 and α-Cx57 were investigated. All five Cx CL2 peptides were found to have high affinity for Ca2+/CaM with dissociation constants (Kd(+Ca)) from 20 to 150 nM. The limiting rate of binding and the rates of dissociation covered a broad range. In addition, we obtained evidence for high affinity Ca2+-independent interaction of all five peptides with CaM, consistent with CaM remaining anchored to gap junctions in resting cells. However, for the α-Cx45 and α-Cx57 CL2 peptides, Ca2+-dependent association at resting [Ca2+] of 50–100 nM is indicated in these complexes as one of the CaM Ca2+ binding sites displays high affinity with Kd of 70 and 30 nM for Ca2+, respectively. Furthermore, complex conformational changes were observed in peptide-apo-CaM complexes with the structure of CaM compacted or stretched by the peptide in a concentration dependent manner suggesting that the CL2 domain may undergo helix-to-coil transition and/or forms bundles, which may be relevant in the hexameric gap junction. We demonstrate inhibition of gap junction permeability by Ca2+/CaM in a dose dependent manner, further cementing Ca2+/CaM as a regulator of gap junction function. The motion of a stretched CaM–CL2 complex compacting upon Ca2+ binding may bring about the Ca2+/CaM block of the gap junction pore by a push and pull action on the CL2 C-terminal hydrophobic residues of transmembrane domain 3 (TM3) in and out of the membrane. [ABSTRACT FROM AUTHOR]

Amyloid β -peptides (A β ) are a major hallmark of Alzheimer's disease (AD) and their neurotoxicity develop with cytosolic calcium dysregulation. On the other hand, calmodulin (CaM), a protein which plays a major multifunctional role in neuronal calcium signaling, has been shown to be involved in the regulation of non-amyloidogenic processing of amyloid β precursor protein (APP). Using fluorescent 6-bromoacetyl-2-dimethylaminonaphthalene derivatives of CaM, Badan-CaM, and human amyloid β (1-42) HiLyte™-Fluor555, we show in this work that A β binds with high affinity to CaM through the neurotoxic A β 25-35 domain. In addition, the affinity of A β for calcium-saturated CaM conformation is approximately 20-fold higher than for CaM conformation in the absence of calcium (apo-CaM). Moreover, the value of K d of 0.98 ± 0.11 nM obtained for A β 1-42 dissociation from CaM saturated by calcium points out that CaM is one of the cellular targets with highest affinity for neurotoxic A β peptides. A major functional consequence of A β -CaM interaction is that it slowdowns A β fibrillation. The novel and high affinity interaction between calmodulin and A β shown in this work opens a yet-unexplored gateway to further understand the neurotoxic effect of A β in different neural cells and also to address the potential of calmodulin and calmodulin-derived peptides as therapeutic agents in AD. [ABSTRACT FROM AUTHOR]

High-resolution ecosystem maps increase the efficiency of policy implementation. However, due to challenges related to both data and methods, such maps of appropriate scale and quality are still rarely available for nationwide analyses. We present solutions to some typical challenges of national-scale ecosystem mapping through the new Ecosystem Map of Hungary. It is a comprehensive, spatially and thematically detailed map with a hierarchical typology. The mapping methodology combined several novel elements from the integration of various large-scale databases in a (theoretical) data cube to the use of image-based predictive mapping (with a Random Forest classifier, using Sentinel-1 and -2 and environmental data). A participatory method involving local experts was used for validation, addressing the lack of suitable reference data as well as improving map-maker and map-user interaction. Besides the original objective of supporting conservation-related decision-making, further uses emerged from a variety of fields including spatial planning, education and recreation. [ABSTRACT FROM AUTHOR]

Abstract: The GL7 epitope was originally described as part of a late lymphocyte activation antigen expressed in mouse and widely used since then as a marker of germinal center. Here we report on its differential expression by rat and human immune cells and lymphoid organs. Expression pattern of the GL7 epitope in rats is similar to that described earlier in mice, namely that GL7 antigen appears only on lymphocytes after 48h activation. In humans lymphocytes, but not the differentiated cells of myeloid origin, express this epitope. The GL7 epitope is up-regulated upon in vitro activation of primary T cells, while a slightly decreased expression is found on B lymphocytes. Fluorescent immunohistochemistry shows discrete location of GL7hi cells in human tonsil. GL7 antibody intensely stains CD19+, IgD+, IgMlow B lymphocytes found at the margin of B cell follicles. The GL7 epitope is constitutively and highly raft-associated in human lymphoid cells. Strong neuraminidase- and partial papain-sensitivity of the GL7 epitope on human lymphocytes indicates a sialic acid-containing epitope linked either to one (or more) membrane protein(s) or to lipids. The lymphocyte-restricted GL7 epitope expression and the activation-dependent bi-directional change in the amount of the epitope suggest a functional role for GL7 epitope linked to carbohydrate-based immunoregulation. [Copyright &y& Elsevier]

Abstract: The involvement of complement in the development and regulation of antibody responses under both healthy and pathological conditions is known for long. Unravelling the molecular mechanisms underlying the events however is still in progress. This review focuses on the role of complement receptors CR1 (CD35) and CR2 (CD21) expressed on T and B cells. Alteration in the expression and function of these receptors may contribute to the initiation and maintenance of immune complex mediated autoimmune diseases such as systemic lupus erythematosus and rheumatoid arthritis. Recent data regarding complement receptor expression on T lymphocytes and on memory B cells are also discussed. [Copyright &y& Elsevier]

Background: Office workers near retirement tend to be sedentary and can be prone to mobility limitations and diseases. We examined the dose effects of exergaming volume and duration of detraining on motor and cognitive function in office workers at late midlife to reduce sedentariness and mobility limitations. Methods: In an assessor-blinded randomized trial, 160 workers aged 55–65 years performed physically active video games in a nonimmersive form of virtual reality (exergaming) in small, supervised groups for 1 h, 1×, 2×, or 3×/week for 8 weeks followed by detraining for 8 and 16 weeks. Exergaming comprises high-intensity, full-body sensorimotor coordination, balance, endurance, and strengthening exercises. The primary outcome was the 6-minute walk test (6MWT), and secondary outcomes were body mass, self-reported physical activity, sleep quality, Berg Balance Scale, Short Physical Performance Battery, fast gait speed, dynamic balance, heart rate recovery after step test, and 6 cognitive tests. Results: The 3 groups were not different in any of the outcomes at baseline (all p > 0.05). The outcomes were stable and had acceptable reliability (intraclass correlation coefficients ≥0.334) over an 8-week control period. Training produced an inverted U-shaped dose response of no (1×), most (2×), and medium (3×/week) effects of exergaming volume in most motor and selected cognitive outcomes. The distance walked in the 6MWT (primary outcome) increased most (94 m, 19%, p < 0.05), medium (57 m, 12%, p < 0.05), and least (4 m, 1%) after exergaming 2×, 3×, or 0× (control) (all different p < 0.05). The highest responders tended to retain the exercise effects over 8 weeks of detraining, independent of training volume. This maintenance effect was less consistent after 16 weeks of detraining. Conclusion: Less was more during training and lasted longer after detraining. A medium dose volume of exergaming produced the largest clinically meaningful improvements in mobility and selected cognitive tests in 60-year-old office workers with mild mobility limitations and intact cognition. [ABSTRACT FROM AUTHOR]

Invasive species are among the main threats to grassland biodiversity, and nature conservation management seeks the best methods to eliminate them and to restore natural habitats. We studied the long‐term effect of mowing on the restoration of Pannonian sand grassland after elimination of invasive black locust (Robinia pseudo‐acacia) plantation in Hungary, Europe. Stands of R. pseudo‐acacia at three sites were felled and stumps herbicide‐treated in the winter of 1994–1995. Mowing with hay removal treatment was applied twice a year in 1995–2001 to assist grassland recovery. A block of 12 adjacent plots of 10 m by 10 m was assigned for the experiment at each site, with six control (unmowed) and six treatment (mowed) plots randomly selected. Vegetation was sampled in June and August yearly in 1995–1999 in all sites, plus in seminatural reference grasslands then re‐sampled six times until 2017 in two sites. Herbicide application with repeated mowing successfully eliminated R. pseudo‐acacia. In the unmowed plots, dense woody cover developed at all sites. Vegetation of mowed plots approached the reference grasslands in sites with better propagule availability of target species according to trajectory analyses. In these sites, higher cover of target species was found in mowed compared to unmowed plots, though still significantly lower than in the reference. Mowed plots were more prone to secondary invasion than the unmowed. The long‐term monitoring revealed that initial mowing assisted the restoration of Pannonian sand grassland, but further management is needed to control secondary invasion and increase target species cover. [ABSTRACT FROM AUTHOR]

Theavailability of seeds of native species in many countries for grassland restoration is limited; therefore, ex situ seed banks can gain importance as a source of germplasm in the future. We tested the usability of seed accessions of the Pannon Seed Bank for reintroduction with the aim to restore sandy grassland in Hungary. Seeds of 10 native sandy grassland species were seeded in the year of collection and after one or two years of storage. The establishment was estimated by counting seedlings along seeded transects for two vegetation seasons. This study produced the first numerical estimate we know about of native sand grassland species emergence in the field. A low establishment of the tested species was found, ranging from 0.002 to 8%. Within this range, Dianthus serotinus had the highest establishment, while Festuca vaginata, which was sown as matrix species, performed only medium establishment. The short‐term storage (1 or 2 years) of seeds had no significant effect, except for F. vaginata, where seed storage had a positive effect on reintroduction success. The year of seeding had the highest influence on recruitment. Four species were found to emerge over two years instead of only the first year. Based on our results, the weak seed yield of certain years and the low supply of native seeds in the market can be mitigated by using stored seeds. It is recommended to use multi‐year, scheduled seeding to reduce the negative impacts of particularly dry years and to increase the restoration success. [ABSTRACT FROM AUTHOR]

Aim: There is increasing recognition that plant–soil feedbacks drive species co‐existence; therefore the below‐ground compartment should be better considered in ecological restoration. Addition of carbon is a restoration measure that relies on indirect plant–soil relationships by immobilizing soil nitrogen in microbial biomass to manipulate competitive hierarchies between plant species. The aim of the present work was to evaluate the impact of six years of carbon amendment in old‐field restoration in the long term, 20 years after the first application. Location: Old‐fields and reference sand steppe on dry calcareous sandy soil in Fülöpháza, Kiskun LTER site (46°53′ N, 19°24′ E), Hungary, Pannonian biogeographic region, Europe. Methods: We applied carbon amendment in the form of sucrose and sawdust on three abandoned agricultural fields between 1998 and 2003. Vegetation was surveyed in 1998–2006 and re‐sampled in 2008, 2010 and 2018 on permanent 2 m × 2 m coenological relevés for carbon‐amended, control and reference plots (n = 144). We used principal response curves (PRC) to describe vegetation development trajectories and linear mixed‐effects models to test changes in cover of vascular plants and cryptogams, nitrogen requirement groups and restoration species groups with time and treatment. Results: Carbon amendment resulted in lower soil nitrogen availability, and lower cover of vascular plants and cryptogams compared to control, but these differences became visible only four to five years after the first application and disappeared three years after the cessation of treatment. Minor treatment effects on the cover of oligotrophic, mesotrophic and target species were found. Conclusions: Carbon amendment did not speed up the recovery of sand grasslands; however, the reduction of cover (vegetation and cryptogam) can be a window of opportunity for other species to colonize that can be used as a complement to other treatments. Long‐term monitoring is especially important in evaluating restoration interventions that focus on indirect above‐ground–below‐ground linkages. [ABSTRACT FROM AUTHOR]

To preserve the natural genetic pattern of species and to avoid the introduction of nonadapted ecotypes during restoration, seed transfer should be spatially restricted. Instead of applying administrative borders in the absence of species‐specific empirical data, biogeographical knowledge can be used as a proxy. Hungary was used as a suitable test region for this approach. The aims of the study were (1) to produce an evidence‐based seed transfer zone (STZ) map applying the Multiple Potential Natural Vegetation model; (2) to assess the uncertainty of the resulting STZ map; and (3) to compare the present seed transfer regulation based on administrative regions with the evidence‐based STZ map. The analysis was based on a floristic map, a vegetation map, and a landscape map of Hungary. Intersected polygons of the three maps were filled with Multiple Potential Natural Vegetation data and clustered to produce seven contiguous units that can serve as STZs. The uncertainty analyses provided a numerical comparison between the two approaches and demonstrated the inadequacy of defining administrative regions as STZs. The practical result of the study is the production of an evidence‐based STZ map that could replace the administrative map currently used for regulation in Hungary. Moreover, this map helps to develop native seed propagation and to enhance ecological restoration. We conclude that field‐based potential vegetation models, similar to the Multiple Potential Natural Vegetation, are suitable for STZ development in countries lacking an evidence‐based system for seed transfer. [ABSTRACT FROM AUTHOR]

Supplemental digital content is available in the text. Introduction: Different therapies can improve clinical and motor symptoms of multiple sclerosis (MS) similarly, but studies comparing the effects of different exercise therapies on clinical and motor outcomes are scant. We compared the effects of exergaming (EXE), balance (BAL), cycling (CYC), proprioceptive neuromuscular facilitation (PNF), and a standard care wait-listed control group (CON) on clinical and motor symptoms and quality of life (QoL) in people with MS (PwMS). Methods: PwMS (n = 68, 90% female; age, 47.0 yr; Expanded Disability Status Scale score 5–6) were randomized into five groups. Before and after the interventions (five times a week for 5 wk), PwMS were tested for MS-related clinical and motor symptoms (Multiple Sclerosis Impact Scale-29 (MSIS-29), primary outcome), QoL (EuroQol Five Dimensions Questionnaire), symptoms of depression, gait and balance ability (Tinetti Assessment Tool), static and dynamic balance and fall risk (Berg Balance Scale), walking capacity (6-min walk test), and standing posturography on a force platform. Results: EXE, BAL, and CYC improved the MSIS-29 scores similarly. EXE and CYC improved QoL and walking capacity similarly but more than BAL. Only EXE improved gait and balance scores (Tinetti Assessment Tool). EXE and BAL improved fall risk and standing balance similarly but more than CYC. PNF and CON revealed no changes. The EuroQol Five Dimensions Questionnaire moderated the exercise effects on the MSIS-29 scores only in EXE. Changes in QoL and changes in the MSIS-29 scores correlated (R 2 = 0.73) only in EXE. Conclusion: In conclusion, BAL and CYC but EXE in particular, but not PNF, can improve clinical and motor symptoms and QoL in PwMS (Expanded Disability Status Scale score 5 to 6), expanding the evidence-based exercise options to reduce mobility limitations in PwMS. [ABSTRACT FROM AUTHOR]

Community assembly theory is suggested as a guiding principle for ecological restoration to help understand the mechanisms that structure biological communities and identify where restoration interventions are needed. We studied three hypotheses related to propagule limitation, stress‐dominance, and limiting similarity concepts in community assembly in a restoration field experiment with a trait‐based null model approach. The experiment aimed to assist the recovery of sand grassland on former arable land in the Kiskunság, Pannonian biogeographic region, Europe. Treatments included initial seeding of five grassland species, carbon amendment, low‐intensity mowing, and combinations in 1 m by 1 m plots in three old fields from 2003 to 2008. The distribution of 10 individual plant traits was compared to the null model and the effect of time and treatments were tested with linear mixed effect models. Initial seeding had the most visible impact on species and trait composition confirming propagule limitation in grassland recovery. Reducing nutrient availability through carbon amendment strengthened trait convergence for length of flowering as expected based on the stress‐dominance hypothesis. Mowing changed trait divergence to convergence for plant height with a strengthening impact with time, supporting our hypothesis of increasing dominance of limiting similarity with time. Our results support the idea that community assembly is simultaneously influenced by propagule limitation and multiple trait‐based processes that act through different traits. The limited impact of manipulating environmental filtering and limiting similarity compared to seeding, however, supports the view that only targeting the dispersal and environmental filters in parallel would improve restoration outcome. [ABSTRACT FROM AUTHOR]

Genetically encoded calcium indicators (GECIs) are useful reporters of cell-signaling, neuronal, and network activities. We have generated novel fast variants and investigated the kinetic mechanisms of two recently developed red-fluorescent GECIs (RGECIs), mApple-based jRGECO1a and mRuby-based jRCaMP1a. In the formation of fluorescent jRGECO1a and jRCaMP1a complexes, calcium binding is followed by rate-limiting isomerization. However, fluorescence decay of calciumbound jRGECO1a follows a different pathway from its formation: dissociation of calcium occurs first, followed by the peptide, similarly to GCaMP-s. In contrast, fluorescence decay of calcium-boundjRCaMP1aoccurs by the reversal of the on-pathway: peptide dissociation is followed by calcium. The mechanistic differences explain the generally slower off-kinetics of jRCaMP1atype indicators compared with GCaMP-s and jRGECO1a-type GECI: the fluorescence decay rate of f-RCaMP1 was 21 s-1, compared with109s-1 forf-RGECO1andf-RGECO2(37 °C). Thus, the CaM-peptide interface is an important determinant of the kinetic responses of GECIs; however, the topology of the structural link to the fluorescent protein demonstrably affects the internal dynamics of the CaM-peptide complex. In the dendrites of hippocampal CA3 neurons, f-RGECO1 indicates calcium elevation in response to a 100 action potential train in a linear fashion, making the probe particularly useful for monitoring large-amplitude, fast signals, e.g. those in dendrites, muscle cells, and immune cells. [ABSTRACT FROM AUTHOR]

Glutamatergic synapses display a rich repertoire of plasticity mechanisms on many different time scales, involving dynamic changes in the efficacy of transmitter release as well as changes in the number and function of postsynaptic glutamate receptors. The genetically encoded glutamate sensor iGluSnFR enables visualization of glutamate release from presynaptic terminals at frequencies up to ~10 Hz. However, to resolve glutamate dynamics during high-frequency bursts, faster indicators are required. Here, we report the development of fast (iGluf) and ultrafast (iGluu) variants with comparable brightness but increased Kd for glutamate (137 μM and 600 μM, respectively). Compared with iGluSnFR, iGluu has a sixfold faster dissociation rate in vitro and fivefold faster kinetics in synapses. Fitting a three-state model to kinetic data, we identify the large conformational change after glutamate binding as the rate-limiting step. In rat hippocampal slice culture stimulated at 100 Hz, we find that iGluu is sufficiently fast to resolve individual glutamate release events, revealing that glutamate is rapidly cleared from the synaptic cleft. Depression of iGluu responses during 100-Hz trains correlates with depression of postsynaptic EPSPs, indicating that depression during high-frequency stimulation is purely presynaptic in origin. At individual boutons, the recovery from depression could be predicted from the amount of glutamate released on the second pulse (paired pulse facilitation/ depression), demonstrating differential frequency-dependent filtering of spike trains at Schaffer collateral boutons. [ABSTRACT FROM AUTHOR]

Improving efficiency of oligonucleotide-directed mutagenesis (ODM) is a prerequisite for wide application of this gene-editing approach in plant science and breeding. Here we have tested histone deacetylase inhibitor treatments for induction of relaxed chromatin and for increasing the efficiency of ODM in cultured maize cells. For phenotypic assay we produced transgenic maize cell lines expressing the non-functional Green Fluorescent Protein ( mGFP) gene carrying a TAG stop codon. These transgenic cells were bombarded with corrective oligonucleotide as editing reagent to recover GFP expression. Repair of green fluorescent protein function was monitored by confocal fluorescence microscopy and flow cytometry was used for quantification of correction events. Sequencing PCR fragments of the GFP gene from corrected cells indicated a nucleotide exchange in the stop codon (TAG) from T to G nucleotide that resulted in the restoration of GFP function. We show that pretreatment of maize cells with sodium butyrate (5-10 mM) and nicotinamide (1-5 mM) as known inhibitors of histone deacetylases can cause elevated chromatin sensitivity to DNase I that was visualized in agarose gels and confirmed by the reduced presence of intact PCR template for the inserted exogenous mGFP gene. Maize cells with more relaxed chromatin could serve as an improved recipient for targeted nucleotide exchange as indicated by an average of 2.67- to 3.62-fold increase in GFP-positive cells. Our results stimulate further studies on the role of the condition of the recipient cells in ODM and testing the application of chromatin modifying agents in other, programmable nuclease-based genome-editing techniques in higher plants. [ABSTRACT FROM AUTHOR]

The expression and role of CR3 (CD11b/CD18) and CR4 (CD11c/CD18) in B cells are not yet explored in contrast to myeloid cells, where these β 2 -integrin type receptors are known to participate in various cellular functions, including phagocytosis, adherence and migration. Here we aimed to reveal the expression and role of CR3 and CR4 in human B cells. In B cells of healthy donors CR3 and CR4 are scarcely expressed. However, two patients with chronic lymphocytic leukemia (CLL) characterized by a peculiar immune-phenotype containing both CD5-positive and CD5-negative B cell populations made possible to study these molecules in distinct B cell subsets. We found that CD11b and CD11c were expressed on both CD5-positive and CD5-negative B cells, albeit to different extents. Our data suggest that these receptors are involved in spreading, since this activity of CpG-activated B cells on fibrinogen could be partially blocked by monoclonal antibodies specific for CD11b or CD11c. CpG-stimulation lead to proliferation of both CD5-positive and CD5-negative B cells of the patients with a less pronounced effect on the CD5-positive cells. In contrast to normal B cells, CLL B cells of both patients reacted to CpG-stimulation with robust IL-10 production. The concomitant, suboptimal stimulus via the BCR and TLR9 exerted either a synergistic enhancing effect or resulted in inhibition of proliferation and IL-10 production of patients’ B cells. Our data obtained studying B cells of leukemic patients point to the role of CR3 and probably CR4 in the interaction of tumor cells with the microenvironment and suggest the involvement of IL-10 producing B cells in the pathologic process. [ABSTRACT FROM AUTHOR]

In order to find the best and most reliable practices for ecological restoration of degraded lands, longer time scales should be considered when evaluating restoration efforts. We assessed the long-term (16 years) effects of different grassland restoration treatments – seeding, mowing, and carbon amendment – in the Pannonian sand grasslands. After re-plowing in 1 m × 1 m plots, treatments were carried out in two abandoned croplands. Seeding was applied only initially (2002) while mowing and carbon amendment were carried out for six years (2003–2008). Vegetation was surveyed yearly from 2003 to 2008 and re-sampled in 2019 in each permanent treatment plot. We used principal coordinates analysis to describe the trajectories of vegetation development and linear mixed-effects models to test changes in the relative cover of native sand grassland (target) and invasive (neophyte) species with time and treatments. Relative cover of target species increased while neophyte species decreased with time in both sites. There was a higher relative cover of target species from the first or third year on and a lower relative cover of neophyte species from the third year on in one site in seeded plots compared to other treatments. Seeded species also spread into non-seeded plots by 2019, obscuring the differences between treatments 16 years after sowing. Carbon amendment proved to be beneficial in the early and mowing in the later phases of restoration. Based on the long-term results, initial seeding is the best method for restoring sand grasslands in old fields by favoring the establishment of target species and controlling non-native invasion. As a supplement to seeding, carbon amendment can be suggested in the initial phases and/or low-intensity mowing in the later phases of the restoration after land abandonment. Although the spread from seeded plots obscured the long-term differences between treatments, it optimized the restoration process, suggesting that the use of small seed introduction units can be enough to restore the whole degraded area. • Long-term monitoring is needed to better interpret restoration interventions. • Early seeding favors target species and controls invasion in grassland restoration. • Small introduction plots can trigger restoration of larger areas. • Low intensity mowing can be applied in the later phases of the restoration process. • Carbon amendment is suggested in combination with seeding right after abandonment. [ABSTRACT FROM AUTHOR]

The application of a filter-based assembly model is a promising concept to support ecological restoration. The filters illustrate dispersal, abiotic and biotic constraints that narrow down the species pool and determine the realized species composition. The aim of restoration interventions is the manipulation of filters to achieve a restoration target., We tested the applicability of a filter-based assembly model in the restoration of endemic Pannonian sandy grassland on old-fields. We report the results of a six-year experiment to identify key filters and find possible interactions. Treatments included the following: (i) seeding (dispersal filter), (ii) carbon amendment to lower nitrogen availability (abiotic filter) and (iii) mowing (biotic filter). Treatments were carried out in 1 m2 plots following the same Latin square design in three old-fields., Seeding was the most important treatment increasing species richness, vascular cover and enhancing target species composition. Mowing played a secondary role, acting primarily in interaction with the other treatments. Carbon amendment significantly reduced soil mineral nitrogen, but played a subordinate role in determining vegetation composition. Significant interactions were found between the biotic and dispersal and the biotic and abiotic filters determining primarily the structural characteristics of the vegetation in terms of vascular cover, moss cover, litter and bare ground. Regarding species composition, synergistic links between the dispersal and biotic filters for seeded species and long-lived forbs, and antagonistic interaction between the dispersal and abiotic filters for seeded species were found., Synthesis and applications. Based on the impact and interactions of filters uncovered in this experiment, we propose a scheme for Pannonian grassland restoration, which can be validated and also used for other oligotrophic systems. In the proposed conceptual model, the dispersal filter plays a key role in determining the outcome of restoration measures followed by the biotic filter if manipulated by mowing, while the abiotic filter altered by nitrogen immobilization proved to have a weak effect. Based on our results, targeting several filters in parallel would improve restoration outcome. We propose that filter interactions should form an integral part of filter-based assembly models and should be taken into account in restoration decisions. [ABSTRACT FROM AUTHOR]

Background: Resting heart rate (RHR) reflects sympathetic nerve activity a significant association between RHR and all-cause and cardiovascular mortality has been reported in some epidemiologic studies. Methods: To analyze the predictive power and accuracy of RHR as a screening measure for individual and clustered cardiovascular risk in adolescents. The study comprised 769 European adolescents (376 boys) participating in the HELENA cross-sectional study (2006–2008) were included in this study. Measurements on systolic blood pressure, HOMA index, triglycerides, TC/HDL-c, VO2máx and the sum of four skinfolds were obtained, and a clustered cardiovascular disease (CVD) risk index was computed. The receiver operating characteristics curve was applied to calculate the power and accuracy of RHR to predict individual and clustered CVD risk factors. Results: RHR showed low accuracy for screening CVD risk factors in both sexes (range 38.5%–54.4% in boys and 45.5%–54.3% in girls). Low specificity’s (15.6%–19.7% in boys; 18.1%–20.0% in girls) were also found. Nevertheless, the sensitivities were moderate-to-high (61.4%–89.1% in boys; 72.9%–90.3% in girls). Conclusion: RHR is a poor predictor of individual CVD risk factors and of clustered CVD and the estimates based on RHR are not accurate. The use of RHR as an indicator of CVD risk in adolescents may produce a biased screening of cardiovascular health in both sexes. [ABSTRACT FROM AUTHOR]

The natural history of introduced species is often unclear due to a lack of historical records. Even when historical information is readily available, important factors of the invasions such as genetic bottlenecks, hybridization, historical relationships among populations and adaptive changes are left unknown. In this study, we developed a set of nuclear, simple sequence repeat markers and used these to characterize the genetic diversity and population structure among native (Eurasian) and non-native (North and South American) populations of Centaurea solstitialis L., (yellow starthistle). We used these data to test hypotheses about the invasion pathways of the species that were based on historical and geographical records, and we make inferences about historical relationships among populations and demographic processes following invasion. We confirm that the center of diversity and the native range of the species is likely the eastern Mediterranean region in the vicinity of Turkey. From this region, the species likely proceeded to colonize other parts of Europe and Asia via a slow, stepwise range expansion. Spanish populations were the primary source of seed to invade South America via human-mediated events, as was evident from historical records, but populations from the eastern Mediterranean region were also important. North American populations were largely derived from South America, but had secondary contributors. We suggest that the introduction history of non-native populations from disparate parts of the native range have allowed not just one, but multiple opportunities first in South America then again in North America for the creation of novel genotypes via intraspecific hybridization. We propose that multiple intraspecific hybridization events may have created especially potent conditions for the selection of a noxious invader, and may explain differences in genetic patterns among North and South America populations, inferred differences in demographic processes, as well as morphological differences previously reported from common garden experiments. [ABSTRACT FROM AUTHOR]

N-Methyl-D-aspartic acid receptor-dependent long term potentiation (LTP), a model of memory formation, requires Ca2+ calmodulin-dependent protein kinase II (aCaMKII) activity and Thr286 autophosphorylation via both global and local Ca2+ signaling, but the mechanisms of signal transduction are not understood. We tested the hypothesis that the Ca2+-binding activator protein calmodulin (CaM) is the primary decoder of Ca2+ signals, thereby determining the output, e.g. LTP. Thus, we investigated the function of CaM mutants, deficient in Ca2+ binding at sites 1 and 2 of the N-terminal lobe or sites 3 and 4 of the C-terminal CaM lobe, in the activation of αCaMKII. Occupancy of CaM Ca2+ binding sites 1, 3, and 4 is necessary and sufficient for full activation. Moreover, the N- and C-terminal CaM lobes have distinct functions. Ca2+ binding to N lobe Ca2+ binding site 1 increases the turnover rate of the enzyme 5-fold, whereas the C lobe plays a dual role; it is required for full activity, but in addition, via Ca2+ binding site 3, it stabilizes ATP binding to aCaMKII 4-fold. Thr286 autophosphorylation is also dependent on Ca2+ binding sites on both the N and the C lobes of CaM. As the CaM C lobe sites are populated by low amplitude/low frequency (global) Ca2+ signals, but occupancy of N lobe site 1 and thus activation of aCaMKII requires high amplitude/high frequency (local) Ca2+ signals, lobe-specific sensing of Ca2+ signaling patterns by CaM is proposed to explain the requirement for both global and local Ca2+ signaling in the induction of LTP via αCaMKII. [ABSTRACT FROM AUTHOR]

Ca2+/calmodulin-dependent protein kinase II (αCaMKII) is thought to exert its role in memory formation by autonomous Ca2+-independent persistent activity conferred by Thr286 autophosphorylation, allowing the enzyme to remain active even when intracellular [Ca2+] has returned to resting levels. Ca2+ sequestration-induced inhibition, caused by a burst of Thr3051306 autophosphorylation via calmodulin (CaM) dissociation from the Thr305/306 sites, is in conflict with this view. The processes of CaM binding, autophosphorylation, and inactivation are dissected to resolve this conflict. Upon Ca2+ withdrawal, CaM sequential domain dissociation is observed, starting with the rapid release of the first (presumed N-terminal) CaM lobe, thought to be bound at the Thy305/306 sites. The time courses of Thr305/306 autophosphorylation and inactivation, however, correlate with the slow dissociation of the second (presumed C-terminal) CaM lobe. Exposure of the Thr305/306 sites is thus not sufficient for their autophosphorylation. Moreover, Thr3051306 autophosphorylation and autoinactivation are shown to occur in the continuous presence of Ca2+ and bound Ca2+/CaM by time courses similar to those seen following Ca2+ sequestration. Our investigation of the activity and mechanisms of phosphoThr286-αCaMKII thus shows time-dependent autoinactivation, irrespective of the continued presence of Ca2+ and CaM, allowing a very short, if any, time window for Ca2+/CaM-free phospho-Thr286-αCaMKII activity. Physiologically, the time-dependent autoinactivation mechanisms of phospho-Thr286αCaMKII (t½ of ∼50 s at 37 °C) suggest a transient kinase activity of ∼1 mm duration in the induction of long term potentiation and thus memory formation. [ABSTRACT FROM AUTHOR]

Studying germination in the native and non-native range of a species can provide unique insights into processes of range expansion and adaptation; however, traits related to germination have rarely been compared between native and non-native populations. In a series of common garden experiments, we explored whether differences in the seasonality of precipitation, specifically, summer drought vs summer rain, and the amount and variation of annual and seasonal precipitation affect the germination responses of populations of an annual ruderal plant, Centaurea solstitialis, from its native range and from two non-native regions with different climates. We found that seeds from all native populations, irrespective of the precipitation seasonality of the region in which they occurred, and non-native populations from regions with dry summers displayed similarly high germination proportions and rates. In contrast, genotypes from the non-native region with predominantly summer rain exhibited much lower germination fractions and rates. Also, percent germination was strongly correlated with variation in precipitation in winter, the season that follows germination for C. solstitialis. Specifically, germination was lower for native and non-native populations experiencing greater variation in winter precipitation. This correlation, however, was greatly influenced by the non-native region with summer rain, which also exhibited the greatest variation in winter precipitation among studied regions. These results suggest that rather than general climatic patterns, the degree of risk experienced at early developmental stages could exert an important control over the germination strategy of C. solstitialis populations in both native and non-native ranges. Also, these findings reveal a largely unique germination response in C. solstitialis genotypes growing in the non-native region with summer rain and high variation in winter precipitation. Our work raises the possibility that rapid adaptive changes in germination strategies may contribute to the success of globally distributed invaders. [ABSTRACT FROM AUTHOR]

Calmodulin plays a key role in the chemical gating of gap junction channels. Two calmodulin-binding regions have previously been identified in connexin32 gap junction protein, one in the N-terminal and another in the C-terminal cytoplasmic tail of the molecule. The aim of this study was to better understand how calmodulin interacts with the connexin32-binding domains. Lobe-specific interactions of calmodulin with connexin32 peptides were studied by stopped flow kinetics, using Ca2+ binding-deficient mutants. Peptides corresponding to the N-terminal tail (residues 1-22) of connexin32 engaged both the N- and C-terminal lobes (N- and C-lobes) of calmodulin, binding with higher affinity to the C-lobe of calmodulin (Ca2+ dissociation rate constants k3,4, 1.7 ± 0.5 s-1) than to the N-lobe (k1,2, 10.8 ± 1.3 s-1). In contrast, peptides representing the C-terminal tail domain (residues 208-227) of connexin32 bound either the Cor the N-lobe but only one calmodulin lobe at a time (k3,4, 2.6 ± 0.1 s-1 or k1, 13.8 ± 0.5 s-1 and k2, 1000 s-1). The calmodulin-binding domains of the N- and C-terminal tails of connexin32 were best defined as residues 1-21 and 216-227, respectively. Our data, showing separate functions of the N- and C-lobes of calmodulin in the interactions with connexin32, suggest transdomain or trans-subunit bridging by calmodulin as a possible mechanism of gap junction gating. [ABSTRACT FROM AUTHOR]

We demonstrate using Ca2+-dependent calmodulin (CaM)-affinity chromatography and overlay with biotinylated CaM that the adaptor proteins growth factor receptor bound (Grb)7 and Grb7V (a naturally occurring variant lacking the Src homology 2 (SH2) domain) are CaM-binding proteins. Deletion of an amphiphilic basic amino-acid sequence (residues 243–256) predicted to form an α-helix located in the proximal region of its pleckstrin homology (PH) domain demonstrates the location of the CaM-binding domain. This site is identical in human and rodents Grb7, and shares great homology with similar regions of Grb10 and Grb14, and the Mig10 protein from Caenorhabditis elegans. We show that Grb7 and Grb7V are present in the cytosol and bound to membranes, while the deletion mutants (Grb7Δ and Grb7VΔ) have less capacity to be associated to membranes. Grb7Δ maintains in part the capacity to bind phosphoinositides, and CaM competes for phosphoinositide binding. Activation of ErbB2 by heregulin β1 decreases the pool of Grb7 associated to membranes. The cell-permeable CaM antagonist W7 (N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide), but not the CaM-dependent protein kinase II inhibitor KN93, prevents this effect. Highly specific cell-permeable CaM inhibitory peptides decrease the association of Grb7 to membranes. This suggests that CaM regulates the intracellular mobilization of Grb7 in living cells. Direct interaction between enhanced yellow fluorescent protein (EYFP)-Grb7 and enhanced cyan fluorescent protein (ECFP)-CaM chimeras at the plasma membrane of living cells was demonstrated by fluorescence resonance energy transfer (FRET). The FRET signal dramatically decreased in cells loaded with a cell-permeable Ca2+ chelator, and was significantly attenuated when enhanced yellow fluorescent protein-Grb7 chimera (EYFP-Grb7)Δ instead of EYFP-Grb7 was used. Finally, we show that conditioned media from cells transiently transfected with Grb7Δ and Grb7VΔ lost its angiogenic activity, in contrast to those from cells transiently transfected with their wild-type counterparts.Oncogene (2005) 24, 4206–4219. doi:10.1038/sj.onc.1208591 Published online 4 April 2005 [ABSTRACT FROM AUTHOR]

Summary Rapid accumulation of toxic products from reactions of reactive oxygen species (ROS) with lipids and proteins significantly contributes to the damage of crop plants under biotic and abiotic stresses. Here we have identified a stress-activated alfalfa gene encoding a novel plant NADPH-dependent aldose/aldehyde reductase that also exhibited characteristics of the homologous human enzyme. The recombinant alfalfa enzyme is active on 4-hydroxynon-2-enal, a known cytotoxic lipid peroxide degradation product. Ectopic synthesis of this enzyme in transgenic tobacco plants provided considerable tolerance against oxidative damage caused by paraquat and heavy metal treatment. These transformants could also resist a long period of water deficiency and exhibited improved recovery after rehydration. We found a reduced production of lipid peroxidation-derived reactive aldehydes in these transformed plants under different stresses. These studies reveal a new and efficient detoxification pathway in plants. [ABSTRACT FROM AUTHOR]

Thr286 autophosphorylation is important for the role of αCaMKII in learning and memory. Phospho-Thr286-αCaMKII has been described to have two types of activity: Ca2+-independent partial activity and Ca2+/calmodulin-activated full activity. We investigated the mechanism of switching between the two activities in order to relate them to the physiological functioning of αCaMKII. Using a fluorometric coupled enzyme assay and smooth muscle myosin light chain (MLC) as substrate, we found that (1) Ca2+-independent activity of phospho-Thr286-αCaMKII represents 5.0 (±3.7)% of the activity measured in the presence of optimal concentrations of Ca2+ and calmodulin and (2) Ca2+ in the presence of calmodulin activates the enzyme with a Km of 137 (±56) nM and a Hill coefficient n = 1.8 (±0.3). In contrast, unphosphorylated αCaMKII has a Km for Ca2+ in the presence of calmodulin of 425 (± 119) nM and a Hill coefficient n = 5.4 (±0.4). Thus, the activity of phospho-Thr286-αCaMKII is essentially Ca2+/ calmodulin dependent with MLC as substrate. In physiological terms, our data suggest that αCaMKII is only activated in stimulated neurones whereas Ca2+/calmodulin activation of phospho-Thr286-αCaMKII can occur in resting cells (∼100 nM {Ca2+}). [ABSTRACT FROM AUTHOR]

Overexpression of the plasma membrane calcium pump (PMCA) isoform 4b by means of the baculovirus system enabled us, for the first time, to study the kinetics of calmodulin binding to this pump. This was done by stopped-flow fluorescence measurements using 2-chloro-(amino-Lys[SUB75])-[6-[4-(N,Ndiethylamino)phenyl]-l,3,5-triazin-4-yl]calmodulin (TA-calmodulin). Upon mixing with PMCA, the fluorescence of TA-calmodulin changed along a biphasic curve: a rapid and small increase in fluorescence was followed by a slow and large decrease that lasted about 100 s. The experiment was done at several PMCA concentrations. Global fitting nonlinear regression analysis of these results led to a model in which PMCA is present in two forms: a closed conformation and an open conformation. Calmodulin reacts with both conformations but reacts faster and with higher affinity for the open conformation. Measurements of the ATPase activity of PMCA under similar conditions revealed that the open form has higher ATPase activity than the closed one. Contrasting with the reaction with the whole pump, TA-calmodulin reacted rapidly (in about 2 s) with a calmodulin-binding peptide made after the sequence of the calmodulinbinding domain of PMCA (C28). Results of TA-calmodulin binding to C28 are explained by a simpler model, in which only an open conformation exists. [ABSTRACT FROM AUTHOR]

Abstract We have used the fluorescently labelled calmodulin TA-CaM to follow calmodulin activation during depolarization of adult rat sensory neurons. Calcium concentration was measured simultaneously using the low affinity indicator Oregon Green BAPTA 5N. TA-CaM fluorescence increased during a 200-ms depolarization but then continued to increase during the subsequent 500 ms, even though total cell calcium was falling at this time. In the next few seconds TA-CaM fluorescence fell, but to a new elevated level that was then maintained for several tens of seconds. During a train of depolarizations that evoked a series of largely independent calcium changes TA-CaM fluorescence was in contrast raised for the duration of the train and for many tens of seconds afterwards. The presence of a peptide corresponding to the calmodulin binding domain of myosin light chain kinase significantly increased the depolarization-induced TA-CaM fluorescence increase and slowed the subsequent fall of fluorescence. We interpret the slow recovery component of the TA-CaM signal as reflecting the slow dissociation of calcium–calmodulin–calmodulin binding protein complexes. Our results show that after brief electrical activity calmodulin's interaction with calmodulin binding proteins persists for approximately one minute. [ABSTRACT FROM AUTHOR]