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Reliable quantitative vegetation reconstructions for Europe during the Holocene are crucial to improving our understanding of landscape dynamics, making it possible to assess the past effects of environmental variables and land-use change on ecosystems and biodiversity, and mitigating their effects in the future. We present here the most spatially extensive and temporally continuous pollen-based reconstructions of plant cover in Europe (at a spatial resolution of 1° × 1°) over the Holocene (last 11.7 ka BP) using the ‘Regional Estimates of VEgetation Abundance from Large Sites’ (REVEALS) model. This study has three main aims. First, to present the most accurate and reliable generation of REVEALS reconstructions across Europe so far. This has been achieved by including a larger number of pollen records compared to former analyses, in particular from the Mediterranean area. Second, to discuss methodological issues in the quantification of past land cover by using alternative datasets of relative pollen productivities (RPPs), one of the key input parameters of REVEALS, to test model sensitivity. Finally, to validate our reconstructions with the global forest change dataset. The results suggest that the RPPs.st1 (31 taxa) dataset is best suited to producing regional vegetation cover estimates for Europe. These reconstructions offer a long-term perspective providing unique possibilities to explore spatial-temporal changes in past land cover and biodiversity.

Our first aim was to document the effects of palaeotemperatures and vegetation changes on beetle assemblages, and secondly to determine the extent to which surrogacy analysis at the family taxonomic level reveals patterns evident from lower taxa analysis. The sedimentary sequence sampled on the experimental site of ‘La Grande Pile’ (Vosges, France) covers the whole of the last climatic cycle. Beetle fragments were extracted from 39 coring samples and identified to the lowest possible taxonomic level. A total of 3092 beetle specimens belonging to 394 taxa were identified, more than half to species level. Carabidae, Staphylinidae and Curculionidae families together represented 40% of the overall taxa richness. Beetle taxa richness and assemblage composition varied markedly over time. Average summer temperatures clearly play a major role in diversity patterns, as temperature was positively correlated with taxon richness. Nevertheless, the warmest and the coldest periods were not the richest and the poorest, respectively, and the most humid period did not correspond to maximum beetle richness. Beetle assemblages are likely to fluctuate in response to other factors such as plant diversity and vegetation structure. Steppe-like vegetation did not reduce species richness while dense, homogenous and closed forests did. Family patterns mirrored those observed at the lower taxa level. This makes the family level a convincing alternative to lower taxonomic level analyses by representing a faithful picture of changing beetle diversity over a long period of time. Finally, evolution of beetle diversity over the Quaternary represents a convincing model for evaluating the effect of close and wide past climate changes, and for assisting in management of present-day biodiversity as part of the current anthropogenic global climate change. [ABSTRACT FROM AUTHOR]

As part of the European Bioanalysis Forum's continued commitment to develop young scientists beyond their scientific skills, we also focus on soft skills and a community responsibility during the Young Scientist Symposia, with the Science Café. In previous years, we have focused on topics such as sustainability (green lab) or the impact of the COVID-19 pandemic on career development. At the ninth Young Scientist Symposium, the Science Café roundtables focused on the work-life balance and how caring for it can be beneficial for both the individual and the company. Feedback from a premeeting survey and from the discussions during the roundtables can be an important addition to personal and professional development. If organizations are not already focusing on the importance of a healthy work-life balance, they can be inspired to include some aspects of the outcome of the Science Café discussions when developing their staff toward future (scientific) leadership.

The extent of vegetation openness in past European landscapes is widely debated. In particular, the temperate forest biome has traditionally been defined as dense, closed-canopy forest; however, some argue that large herbivores maintained greater openness or even wood-pasture conditions. Here, we address this question for the Last Interglacial period (129,000-116,000 years ago), before Homo sapiens -linked megafauna declines and anthropogenic landscape transformation. We applied the vegetation reconstruction method REVEALS to 96 Last Interglacial pollen records. We found that light woodland and open vegetation represented, on average, more than 50% cover during this period. The degree of openness was highly variable and only partially linked to climatic factors, indicating the importance of natural disturbance regimes. Our results show that the temperate forest biome was historically heterogeneous rather than uniformly dense, which is consistent with the dependency of much of contemporary European biodiversity on open vegetation and light woodland.

Macrocyclization constraints are widely used in the design of protein ligands to stabilize their bioactive conformation and increase their affinities. However, the resulting changes in binding entropy can be puzzling and uncorrelated to affinity gains. Here, the thermodynamic (Isothermal Titration Calorimetry) and structural (X-ray, NMR and CD) analysis of a complete series of lactam-bridged peptide ligands of the vascular endothelial growth factor, and their unconstrained analogs are reported. It is shown that differences in thermodynamics arise mainly from the folding energy of the peptide upon binding. The systematic reduction in conformational entropy penalty due to helix pre-organization can be counterbalanced by an unfavorable vibrational entropy change if the constraints are too rigid. The gain in configurational entropy partially escapes the enthalpy/entropy compensation and leads to an improvement in affinity. The precision of the analytical ITC method makes this study a possible benchmark for constrained peptides optimization., (© 2022 The Authors. Chemistry - A European Journal published by Wiley-VCH GmbH.)

The thiol group of the cysteine side chain is arguably the most versatile chemical handle in proteins. To expand the scope of established and commercially available thiol bioconjugation reagents, we genetically encoded a second such functional moiety in form of a latent thiol group that can be unmasked under mild physiological conditions. Phenylacetamidomethyl (Phacm) protected homocysteine (HcP) was incorporated and its latent thiol group unmasked on purified proteins using penicillin G acylase (PGA). The enzymatic deprotection depends on steric accessibility, but can occur efficiently within minutes on exposed positions in flexible sequences. The freshly liberated thiol group does not require treatment with reducing agents. We demonstrate the potential of this approach for protein modification with conceptually new schemes for regioselective dual labeling, thiol bioconjugation in presence of a preserved disulfide bond and formation of a novel intramolecular thioether crosslink., (© 2021 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.)

A multidisciplinary study (geomorphology, sedimentology and palynology) shows that the landscapes of the southwest coast of Corsica have been deeply modified by humans and the climate since 3000 BC. Significant and rapid landscape transformations are recorded between the Chalcolithic and the Middle Bronze Ages (3000-1300 BC). Several major (2.2 ka BC, 1.2 ka BC) and local (3000 BC) detrital events affected the Taravo Lower Valley in relation to global climatic changes and anthropic activities. The vegetation dynamics since 3000 BC show alternating phases of agriculture and abandonment until the complete disappearance of the original forest populations in the vicinity of the Canniccia Marshes. An early phase of Olea cultivation is recorded between 2900 and 2300 BC. Plant macro-remains indicate that cereals, vine and many species of Fabaceae were cultivated in the nearby of the archaeological sites during the middle to the late Chalcolithic Age. The event of 2.2 ka BC corresponds to an abandonment phase in the lower Taravo Valley. Pastoralism dominated agricultural activities between 2200 and 1700 BC. During Roman times, agriculture is characterized by olive and vine cultivation. A new peak of pastoralism and the cultivation of Castanea are noted during invasion times (500 to 1000 AD), showing that invasions didn't disturb agricultural activities in the Taravo Valley. During the Pisa Period (end of the 9th C. to then end of 13th C. AD), pastoralism declined and vine and cereals were cultivated in the very nearby of the Canniccia Marshes. During the Genoa Period upwards (end of the 13th C. to 1769 AD), a decline in agriculture and a recrudescence of the forest (maquis and pine) are recorded, leading to the settlement of a present-day vegetal landscape dominated by an Erica arborea maquis., Competing Interests: The authors have declared that no competing interests exist.

We report the genetically encoded incorporation of phenylacetyl protected lysine (PacK) into proteins in Escherichia coli. This unnatural side-chain modification can be enzymatically removed using either penicillin G acylase (PGA) or, surprisingly, the sirtuin SrtN from Bacillus subtilis. Our approach expands the toolbox to reversibly control protein structure and function under very mild and non-denaturing conditions, as demonstrated by triggering the activity of the nonribosomal peptide synthetase GrsA.

The interaction between vascular endothelial growth factor (VEGF) and its receptors (VEGFR) has important implications in angiogenesis and cancer, which moved us to search for peptide derivatives able to block this protein-protein interaction. In a previous work we had described a collection of linear 13-mer peptides specially designed to adopt helical conformations (Ac-SSEE X⁵ ARN X⁸ AA X 12 N-NH₂), as well as the evaluation of seven library components for the inhibition of the interaction of VEGF with its Receptor 1 (VEGFR1). This study led to the discovery of some new, quite potent inhibitors of this protein-protein system. The results we found prompted us to extend the study to other peptides of the library. We describe here the evaluation of a new selection of peptides from the initial library that allow us to identify new VEGF-VEGFR1 inhibitors. Among them, the peptide sequence containing F, W, and I residues at the 5, 9, and 12 positions, show a very significant nanomolar IC 50 value, competing with VEGF for its receptor 1, VEGFR1 (Flt-1), which could represent a new tool within the therapeutic arsenal for cancer detection and therapy., Competing Interests: The authors declare no conflict of interest.

Cancer angiogenesis is mainly initiated by vascular endothelial growth factors (VEGFs). On the basis of the reported crystal structures of three natural ligands (VEGF-A, -B, and PlGF) with the major receptors VEGFR-1 and VEGFR-2, we scanned receptor-binding epitopes of these ligands by designing linear and cyclic peptides with the aim to disrupt the VEGF-A/VEGFR-1 interaction, which is implicated in cancer development. The ability of peptides to inhibit this interaction was evaluated by an ELISA-based assay. Several peptides, especially those mimicking loop 1 (L1) of these ligands that binds primarily to domain D3 of VEGFRs, have demonstrated higher inhibition for VEGF-A/VEGFR-1 binding. They have also shown inhibitory effects on VEGF-induced tube formation in HUVECs (human umbilical vein endothelial cells). These results validate the domain D3 of VEGFRs as an efficient target for the design of VEGFR antagonists.

The VEGFR1 has been shown to play a role in the regulation of angiogenesis, and has therefore been associated to several pathologies. In order to extend our toolbox of screening methods for the identification of compounds disrupting the VEGF receptor 1/VEGF interaction, we developed a fast and accurate displacement assay, in which VEGF receptor 1 domain 2 is directly labeled with an enzyme, bypassing the classical streptavidin-biotin interaction system. A description of this straightforward strategy is provided here, including its advantages and disadvantages. Optimization of the reagents preparation, purification and conservation, and displacement assay with known molecular entities are presented., (Copyright © 2017. Published by Elsevier Inc.)

Angiogenesis is tightly regulated through the binding of vascular endothelial growth factors (VEGFs) to their receptors (VEGFRs). In this context, we showed that human VEGFR1 domain 2 crystallizes in the presence of Zn2+, Co2+ or Cu2+ as a dimer that forms via metal-ion interactions and interlocked hydrophobic surfaces. SAXS, NMR and size exclusion chromatography analyses confirm the formation of this dimer in solution in the presence of Co2+, Cd2+ or Cu2+. Since the metal-induced dimerization masks the VEGFs binding surface, we investigated the ability of metal ions to displace the VEGF-A binding to hVEGFR1: using a competition assay, we evidenced that the metals displaced the VEGF-A binding to hVEGFR1 extracellular domain binding at micromolar level., Competing Interests: The authors have declared that no competing interests exist.

The v114* cyclic peptide has been identified as a tight vascular endothelial growth factor (VEGF) ligand. Here we report on the use of isothermal titration calorimetry (ITC), 96-well plate competition assay, and circular dichroism (CD) to explore the binding determinants of a new set of related peptides. Anti-VEGF antibodies are currently used in the clinic for regulating angiogenesis in cancer and age-related macular degeneration treatment. In this context, our aim is to develop smaller molecular entities with high affinity for the growth factor by a structure activity relationship approach. The cyclic disulfide peptide v114* was modified in several ways, including truncation, substitution, and variation of the size and nature of the cycle. The results indicated that truncation or substitution of the four N-terminal amino acids did not cause severe loss in affinity, allowing potential peptide labeling. Increase of the cycle size or substitution of the disulfide bridge with a thioether linkage drastically decreased the affinity, due to an enthalpy penalty. The leucine C-terminal residue positively contributed to affinity. Cysteine N-terminal acetylation induced favorable ΔΔG° and ΔΔH° of binding, which correlated with free peptide CD spectra changes. We also propose a biochemical model to extrapolate Ki from IC50 values measured in the displacement assay. These calculated Ki correlate well with the Kd values determined by extensive direct and reverse ITC measurements.

Previously designed cyclic peptide antagonist c[YYDEGLEE]-NH2 disrupts the interaction between vascular endothelial growth factor (VEGF) and its receptors (VEGFRs). It represents a promising tool in the fight against cancer and age-related macular degeneration. We described in this paper the optimization of the lead peptide by C-terminal modification. A new strategy for the synthesis of cyclic peptides is developed, improving the cyclisation efficiency. At 100 µM, several new peptides with an aromatic group flexibly linked at C-terminal end showed significantly increased receptor binding affinities in competition ELISA test. The most active peptide carrying a coumarin group may be a useful tool in anti-angiogenic biological studies.

Objectives: We studied whether plasma levels of angiogenic factors VEGF and placental growth factor (PlGF) in coronary artery disease patients or undergoing cardiac surgery are modified, and whether those factors modulate endothelial progenitor's angiogenic potential., Methods and Results: A total of 143 patients' plasmas from two different studies were analyzed (30 coronary artery disease patients, 30 patients with stable angina, coupled with 30 age and sex-matched controls; 53 patients underwent cardiac surgery). Among factors screened, only PlGF was found significantly increased in these pathological populations. PlGF-1 and PlGF-2 were then tested on human endothelial-colony-forming cells (ECFCs). We found that PlGF-1 and PlGF-2 induce VEGFR1 phosphorylation and potentiate ECFCs tubulogenesis in vitro. ECFCs VEGFR1 was further inhibited using a specific small interfering RNA (siRNA) and the chemical compound 4321. We then observed that the VEGFR1-siRNA and the compound 4321 decrease ECFCs tubulogenesis potential in vitro. Finally, we tested the compound 4321 in the preclinical Matrigel(®)-plug model with C57Bl/6J mice as well as in the murine hindlimb ischemia model. We found that 4321 inhibited the plug vascularization, attested by the hemoglobin content and the VE-Cadherin expression level and that 4321 inhibited the post-ischemic revascularization., Conclusion: PlGF plasma levels were found increased in cardiovascular patients. Disrupting PlGF/VEGFR1 pathway could modulate ECFC-induced tubulogenesis, the cell type responsible for newly formed vessels in vivo.

Protein-protein interactions (PPIs) have emerged as important targets for pharmaceutical intervention because of their essential role in numerous physiological and pathological processes, but screening efforts using small-molecules have led to very low hit rates. Linear peptides could represent a quick and effective approach to discover initial PPI hits, particularly if they have inherent ability to adopt specific peptide secondary structures. Here, we address this hypothesis through a linear helical peptide library, composed of four sublibraries, which was designed by theoretical predictions of helicity (Agadir software). The 13-mer peptides of this collection fixes either a combination of three aromatic or two aromatic and one aliphatic residues on one face of the helix (Ac-SSEEX(5)ARNX(9)AAX(12)N-NH2), since these are structural features quite common at PPIs interfaces. The 81 designed peptides were conveniently synthesized by parallel solid-phase methodologies, and the tendency of some representative library components to adopt the intended secondary structure was corroborated through CD and NMR experiments. As proof of concept in the search for PPI modulators, the usefulness of this library was verified on the widely studied p53-MDM2 interaction and on the communication between VEGF and its receptor Flt-1, two PPIs for which a hydrophobic α-helix is essential for the interaction. We have demonstrated here that, in both cases, selected peptides from the library, containing the right hydrophobic sequence of the hot-spot in one of the protein partners, are able to interact with the complementary protein. Moreover, we have discover some new, quite potent inhibitors of the VEGF-Flt-1 interaction, just by replacing one of the aromatic residues of the initial F(5)Y(9)Y(12) peptide by W, in agreement with previous results on related antiangiogenic peptides. Finally, the HTS evaluation of the full collection on thermoTRPs has led to a few antagonists of TRPV1 and TRPA1 channels, which open new avenues on the way to innovative modulators of these channels.

The design, synthesis, conformational studies and binding affinity for VEGF receptors of a collection of linear and cyclic peptide analogues of the N-terminal α-helix fragments 13-25 of VEGF and 1-13 of Vammin are described. Linear 13(14)-mer peptides were designed with the help of an AGADIR algorithm and prepared following peptide solid-phase synthetic protocols. Cyclic peptide derivatives were prepared on-resin from linear precursors with conveniently located Glu and Lys residues, by the formation of amide linkages. Conformational analysis, CD and NMR, showed that most synthesized peptides have a clear tendency to be structured as α-helices in solution. Some of the peptides were able to bind a VEGFR-1 receptor with moderate affinity. In addition to the described key residues (Phe17, Tyr21 and Tyr25), Val14 and Val20 seem to be relevant for affinity.

The study of two pollen sequences from El-Kala marshes allowed the reconstruction of the regional vegetation history supported by eight radiocarbon dates. Pollen assemblages from Bourdim site were dominated by local input of Alnus and Salix, while regional vegetation was characterized by scattered Quercus suber forests with a well-developed Erica arborea matorral. While the vegetation dynamics recorded at Bourdim is recent (Late Holocene), the majority of the pollen diagram from Garaat El-Ouez is contemporaneous to the Late Pleniglacial and is characterized by open woodlands with Pinus, Poaceae and several heliophilous herbs. The significant values of Cedrus pollen identified in this period indicate that the region of El-Kala most probably played the role of a refugium for this tree., (Copyright © 2010 Académie des sciences. Published by Elsevier SAS. All rights reserved.)

The Velay sequence (France) provides a unique, continuous, palynological record spanning the last four climatic cycles. A pollen-based reconstruction of temperature and precipitation displays marked climatic cycles. An analysis of the climate and vegetation changes during the interglacial periods reveals comparable features and identical major vegetation successions. Although Marine Isotope Stage (MIS) 11.3 and the Holocene had similar earth precessional variations, their correspondence in terms of vegetation dynamics is low. MIS 9.5, 7.5, and especially 5.5 display closer correlation to the Holocene than MIS 11.3. Ecological factors, such as the distribution and composition of glacial refugia or postglacial migration patterns, may explain these discrepancies. Comparison of ecosystem dynamics during the past five interglacials suggests that vegetation development in the current interglacial has no analogue from the past 500,000 years.

The last interglacial period (127-110 kyr ago) has been considered to be an analogue to the present interglacial period, the Holocene, which may help us to understand present climate evolution. But whereas Holocene climate has been essentially stable in Europe, variability in climate during the last interglacial period has remained unresolved, because climate reconstructions from ice cores, continental records and marine sediment cores give conflicting results for this period. Here we present a high-resolution multi-proxy lacustrine record of climate change during the last interglacial period, based on oxygen isotopes in diatom silica, diatom assemblages and pollen-climate transfer functions from the Ribains maar in France. Contrary to a previous study, our data do not show a cold event interrupting the warm interglacial climate. Instead, we find an early temperature maximum with a transition to a colder climate about halfway through the sequence. The end of the interglacial period is clearly marked by an abrupt change in all proxy records. Our study confirms that in southwestern Europe the last interglacial period was a time of climatic stability and is therefore still likely to represent a useful analogue for the present climate.

Two Holocene lake sequences from Lac de Creno, Corsica were analysed on the basis of 119 pollen spectra and with the support of 13 14 C-calibrated dates. The lower part of one of these sequences, corresponding to the late- glacial period, has been published previously. The first third of the Post-glacial is characterized by very particular forest dynamics, namely the absence of a role for deciduous Quercus and Corylus, the presence of mesophilous vegetation types dominated by Taxus, and the major forest role of Erica arborea at lower and mean altitude. At about 7440 cal BP, the occurrence of a major anthropogenic action brought about significant changes in the vegetation, notably an increase of deciduous Quercus and the expansion of Quercus ilex. Later, three major human-induced events are identified: the first, at about 2290 cal BP, is the cause of a short local expansion of Abies; the second, at about 1150 cal BP, is the degradation of deciduous forests to the benefit of Fagus; the third, at about 310 cal BP, corresponds to the disappearance of Fagus and its replacement by Pinus. Pollen data indicate that Q. ilex, Abies and Fagus are not indigenous in Corsica but spread there during the Postglacial ; this probably took place at about 6980 cal BP for Q. ilex.

The pollen-analytical study of eight sites (two glacial lakes and six coastal marshes) has been carried out (nine pollen diagrams, 472 pollen spectra). The total absence of Quercus ilex forest in Corsica during the Atlantic and the vegetation contrast existing at that time between the western and the eastern sides of the island are demonstrated. On the western site, Erica arborea constituted the only regional vegetation from the coast up to about 1500 m. Human action is responsible for the establishment of Quercus ilex in this climax vegetation during the Sub-boreal. It is also shown that the only Pinus halepensis forest present today in Corsica has been introduced on the island during the nineteenth century.