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Introduction: Effective infiltration of chimeric antigen receptor T (CAR-T) cells into solid tumors is critical for achieving a robust antitumor response and improving therapeutic outcomes. While CAR-T cell therapies have succeeded in hematologic malignancies, their efficacy in solid tumors remains limited due to poor tumor penetration and an immunosuppressive tumor microenvironment. This study aimed to evaluate the potential of low-dose radiotherapy (LDRT) administered before T-cell therapy to enhance the antitumor effect by promoting CAR-T cell infiltration. We hypothesized that combining LDRT with T-cell therapy would improve tumor control and survival compared to either treatment alone., Methods: We investigated this hypothesis using two NSG mouse models bearing GSU or CAPAN-2 solid tumors. The mice were treated with engineered CAR-T cells targeting guanyl cyclase-C (GCC) or mesothelin as monotherapy or in combination with LDRT. Additionally, we extended this approach to a C57BL/6 mouse model implanted with MC38-gp100+ cells, followed by adoptive transfer of pmel+ T cells before and after LDRT. Tumor growth and survival outcomes were monitored in all models. Furthermore, we employed atomic force microscopy (AFM) in a small cohort to assess the effects of radiotherapy on tumor stiffness and plasticity, exploring the role of tumor nanomechanics as a potential biomarker for treatment efficacy., Results: Our results demonstrated enhanced tumor control and prolonged survival in mice treated with LDRT followed by T-cell therapy across all models. The combination of LDRT with CAR-T or pmel+ T-cell therapy led to superior tumor suppression and survival compared to monotherapy, highlighting the synergistic impact of the combined approach. Additionally, AFM analysis revealed significant changes in tumor stiffness and plasticity in response to LDRT, suggesting that the nanomechanical properties of the tumor may be predictive of therapeutic response., Discussion: The findings of this study highlight the transformative potential of incorporating LDRT as a precursor to adoptive T-cell therapy in solid tumors. By promoting CAR-T and pmel+ T-cell infiltration into the tumor microenvironment, LDRT enhanced tumor control and improved survival outcomes, offering a promising strategy to overcome the challenges associated with CAR-T therapy in solid tumors. Additionally, the changes in tumor nanomechanics observed through AFM suggest that tumor stiffness and plasticity could be biomarkers for predicting treatment outcomes. These results support further investigation into the clinical application of this combined approach to improve the efficacy of cell-based therapies in patients with solid tumors., Competing Interests: JWW: Accuray (SAB, consulting), Alpine Immune Science (SAB, consulting, equity), Boehringer Ingelheim (SAB, consulting), Checkmate Pharmaceuticals (SAB, consulting, equity), China Medical Tribune (SAB, consulting), Genentech (SAB, consulting), GI Innovation (SAB, consulting), Kezar Life Sciences (consulting, SAB), Legion Healthcare Partners (SAB, consulting), Life Science Dynamic Limited (SAB, consulting), McKesson Corporation (SAB, consulting), Molecular Match (equity), Nanorobotix (SAB, consulting), OligoImmune (founder), Roche (SAB, consulting), Roche Molecular Systems (SAB, consulting), Nanobiotix (Research grant, Travel expenses, SAB), BMS (Research grant), Merck (Research grant), Varian (Research grant, Travel expenses, clinical sponsored research), Reflexion (Research grant, Travel expenses, Stock options, SAB), Hotspot Therapeutics (Research grant), Gilead (Research grant), Novocure (SAB), Oncoresponse (SAB, Stock options), Astra Zeneca (consultant, Research grant), Bayer Healthcare (Research grant), Kiromic (Research grant), Alkermes (Research grant, SAB), Artidis (Research grant), Sciclone (Research grant), Takeda (Research grant), Pebble Life Science (Research grant). HB and NP-O report consulting support from Research Guided Cancer Treatment Consults. Authors KX, CW and JM were employed by Takeda Development Centers Americas Inc. Authors GS, SN, PO, ML and MP were employed by ARTIDIS AG. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision. The authors declare that this study received funding from Takeda and was involved in the study design. The authors declare that ARTIDIS funded the tissue nanomechanical analysis from the study funded entirely by Takeda. ARTIDIS personnel was involved in the collection, analysis, interpretation, and writing of the tissue nanomechanical section of this manuscript., (Copyright © 2024 Puebla-Osorio, Fowlkes, Barsoumian, Xega, Srivastava, Kettlun-Leyton, Nizzero, Voss, Riad, Wong, Huang, Hu, Mitchell, Kim, Rafiq, He, Sezen, Hsu, Masrorpour, Maleki, Leuschner, Cortez, Oertle, Loparic, Plodinec, Markman and Welsh.)

With ongoing global warming, increasing water deficits promote physiological stress on forest ecosystems with negative impacts on tree growth, vitality, and survival. How individual tree species will react to increased drought stress is therefore a key research question to address for carbon accounting and the development of climate change mitigation strategies. Recent tree-ring studies have shown that trees at higher latitudes will benefit from warmer temperatures, yet this is likely highly species-dependent and less well-known for more temperate tree species. Using a unique pan-European tree-ring network of 26,430 European beech (Fagus sylvatica L.) trees from 2118 sites, we applied a linear mixed-effects modeling framework to (i) explain variation in climate-dependent growth and (ii) project growth for the near future (2021-2050) across the entire distribution of beech. We modeled the spatial pattern of radial growth responses to annually varying climate as a function of mean climate conditions (mean annual temperature, mean annual climatic water balance, and continentality). Over the calibration period (1952-2011), the model yielded high regional explanatory power (R 2  = 0.38-0.72). Considering a moderate climate change scenario (CMIP6 SSP2-4.5), beech growth is projected to decrease in the future across most of its distribution range. In particular, projected growth decreases by 12%-18% (interquartile range) in northwestern Central Europe and by 11%-21% in the Mediterranean region. In contrast, climate-driven growth increases are limited to around 13% of the current occurrence, where the historical mean annual temperature was below ~6°C. More specifically, the model predicts a 3%-24% growth increase in the high-elevation clusters of the Alps and Carpathian Arc. Notably, we find little potential for future growth increases (-10 to +2%) at the poleward leading edge in southern Scandinavia. Because in this region beech growth is found to be primarily water-limited, a northward shift in its distributional range will be constrained by water availability., (© 2024 The Author(s). Global Change Biology published by John Wiley & Sons Ltd.)

Recent findings suggest that immunoradiotherapy (IRT), combining photon radiotherapy (XRT) or proton radiotherapy (PRT) with immune checkpoint blockade, can enhance systemic tumor control. However, the comparative efficacy of XRT and PRT in IRT remains understudied. To address this, we compared outcomes between XRT + αPD1 and PRT + αPD1 in murine αPD1-resistant lung cancer (344SQR). We also assessed the impact of the nanoparticle radioenhancer NBTXR3 on both XRT + αPD1 and PRT + αPD1 for tumor control and examined the tumor immune microenvironment using single-cell RNA sequencing (scRNAseq). Additionally, mice cured by NBTXR3 + PRT + αPD1 were rechallenged with three lung cancer cell lines to evaluate memory antitumor immunity. PRT + αPD1 showed superior local tumor control and abscopal effects compared to XRT + αPD1. NBTXR3 + PRT + αPD1 significantly outperformed NBTXR3 + XRT + αPD1 in tumor control, promoting greater infiltration of antitumor lymphocytes into irradiated tumors. Unirradiated tumors treated with NBTXR3 + PRT + αPD1 had more NKT cells, CD4 T cells, and B cells, with fewer Tregs, than those treated with NBTXR3 + XRT + αPD1. NBTXR3 + PRT + αPD1 also stimulated higher expression of IFN-γ, GzmB, and Nkg7 in lymphocytes, reduced the TGF-β pathway, and increased tumor necrosis factor alpha expression compared to NBTXR3 + XRT + αPD1. Moreover, NBTXR3 + PRT + αPD1 resulted in greater M1 macrophage polarization in both irradiated and unirradiated tumors. Mice achieving remission through NBTXR3 + PRT + αPD1 exhibited a robust memory immune response, effectively inhibiting growth of subsequent tumors from three distinct lung cancer cell lines. Proton IRT combined with NBTXR3 offers enhanced tumor control and survival rates over photon-based treatments in managing αPD1-resistant lung cancer, indicating its potential as a potent systemic therapy., (© 2024. The Author(s).)

Recent hot droughts have caused tree vitality decline and increased mortality in many forest regions on earth. Most of Central Europe's important timber species have suffered from the extreme 2018/2019 hot drought, confronting foresters with difficult questions about the choice of more drought- and heat-resistant tree species. We compared the growth dynamics of European beech, sessile oak, Scots pine and Douglas fir in a warmer and a cooler lowland region of Germany to explore the adaptive potential of the four species to climate warming (24 forest stands). The basal area increment (BAI) of the two conifers has declined since about 1990-2010 in both regions, and that of beech in the warmer region, while oak showed positive BAI trends. A 2 °C difference in mean temperatures and a higher frequency of hot days (temperature maximum >30 °C) resulted in greater sensitivity to a negative climatic water balance in beech and oak, and elevated sensitivity to summer heat in Douglas fir and pine. This suggests to include hot days in climate-growth analyses. Negative pointer years were closely related to dry years. Nevertheless, all species showed growth recovery within one to three years. We conclude that all four species are sensitive to a deteriorating climatic water balance and hot temperatures, and have so far not been able to successfully acclimate to the warmer climate, with especially Douglas and beech, but also Scots pine, being vulnerable to a warming and drying climate., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

The future performance of the widely abundant European beech (Fagus sylvatica L.) across its ecological amplitude is uncertain. Although beech is considered drought-sensitive and thus negatively affected by drought events, scientific evidence indicating increasing drought vulnerability under climate change on a cross-regional scale remains elusive. While evaluating changes in climate sensitivity of secondary growth offers a promising avenue, studies from productive, closed-canopy forests suffer from knowledge gaps, especially regarding the natural variability of climate sensitivity and how it relates to radial growth as an indicator of tree vitality. Since beech is sensitive to drought, we in this study use a drought index as a climate variable to account for the combined effects of temperature and water availability and explore how the drought sensitivity of secondary growth varies temporally in dependence on growth variability, growth trends, and climatic water availability across the species' ecological amplitude. Our results show that drought sensitivity is highly variable and non-stationary, though consistently higher at dry sites compared to moist sites. Increasing drought sensitivity can largely be explained by increasing climatic aridity, especially as it is exacerbated by climate change and trees' rank progression within forest communities, as (co-)dominant trees are more sensitive to extra-canopy climatic conditions than trees embedded in understories. However, during the driest periods of the 20th century, growth showed clear signs of being decoupled from climate. This may indicate fundamental changes in system behavior and be early-warning signals of decreasing drought tolerance. The multiple significant interaction terms in our model elucidate the complexity of European beech's drought sensitivity, which needs to be taken into consideration when assessing this species' response to climate change., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

The persistence of plant populations depends crucially on successful regeneration. Yet, little is known about the effects of consecutive winter and spring frost events on the regeneration stage of trees from different seed sources, although this will partly determine the success of climate warming-driven poleward range shifts. In a common garden experiment with European beech ( Fagus sylvatica ) seedlings from winter 2015/2016 to autumn 2017, we studied how simulated successive spring and winter frost events affect leaf-out dates, growth performance, and survival rates of 1- to 2-year-old seedlings from provenances differing in climate at origin. We further investigated the combined effects of successive frost events. The first spring frost after germination led to a mortality rate up to 75%, resulting in reduced seedling numbers but better frost tolerance of the survivors, as reflected in a weaker impact of the following winter frost event in the survivors compared to the non-acclimated control. Final plant height was most strongly reduced by the spring frost in the second year. The winter frost event delayed leaf-out by up to 40 days, leading to severe growth impairment in 2017. Our results indicate partly successful frost acclimation and/or the selection of frost-hardier individuals, because the negative growth effects of consecutive frost events did not add up after exposure to more than one event. Both mechanisms may help to increase the frost tolerance of beech offspring. Nevertheless, mortality after the first spring frost was high, and frost exposure generally caused growth reductions. Thus, achieving higher frost tolerance may not be sufficient for beech seedlings to overcome frost-induced reductions in competitive strength caused by winter frost damage and delayed leaf enfolding., Competing Interests: All authors have no conflicts of interest to declare., (© 2024 The Author(s). Ecology and Evolution published by John Wiley & Sons Ltd.)

To increase the resilience of forests to drought and other hazards, foresters are increasingly planting mixed stands. This requires knowledge about the drought response of tree species in pure and mixed-culture neighborhoods. In addition, drought frequently interacts with continued atmospheric nitrogen (N) deposition. To disentangle these factors for European beech, Norway spruce and Douglas fir, we conducted a replicated 3-factorial sapling growth experiment with three moisture levels, (high, medium, and low), two N levels (high and ambient), and pure and mixed-culture neighborhoods. We measured biomass, stomatal conductance (GS), shoot water potential (at predawn: ΨPD, midday, and turgor loss point: ΨTLP), branch xylem embolism resistance (Ψ50) and minimum epidermal conductance (Gmin). The three species differed most with respect to Gmin (10-fold higher in beech than in the conifers), hydroscape area (larger in beech), and the time elapsed to reach stomatal closure (TΨGS90) and ΨTLP (TTLP; shorter in beech), while Ψ50 and ΨTLP were remarkably similar. Neighborhood (pure vs mixed-culture) influenced biomass production, water status and hydraulic traits, notably GS (higher in Douglas fir, but lower in spruce and beech, in mixtures than pure culture), hydraulic safety margin (smaller for beech in mixtures), and TΨGS90 and TTLP (shorter for spruce in mixture). High N generally increased GS, but no consistent N effects on leaf water status and hydraulic traits were detected, suggesting that neighbor identity had a larger effect on plant water relations than N availability. We conclude that both tree neighborhood and N availability modulate the drought response of beech, spruce, and Douglas fir. Species mixing can alleviate the drought stress of some species, but often by disadvantaging other species. Thus, our study suggests that stabilizing and building resilience of production forests against a drier and warmer climate may depend primarily on the right species choice; species mixing can support the agenda., (© Crown copyright 2024.)

Fine roots are multifunctional organs that may change function with ageing or root branching events from primarily absorptive to resource transport and storage functions. It is not well understood, how fine root branching patterns and related root functional differentiation along the longitudinal root axis change with soil chemical and physical conditions. We examined the variation in fine root branching patterns (the relative frequency of 1st to 4th root orders) and root morphological and chemical traits of European beech trees with soil depth (topsoil vs. subsoil) and soil chemistry (five sites with acid to neutral/alkaline bedrock). Bedrock type and related soil chemistry had an only minor influence on branching patterns: base-poor, infertile sites showed no higher fine root branching than base-rich sites. The contribution of 1st-order root segments to total fine root length decreased at all sites from about 60% in the topsoil (including organic layer) to 45% in the lower subsoil. This change was associated with a decrease in specific root area and root N content and an increase in mean root diameter with soil depth, while root tissue density did not change consistently. We conclude that soil depth (which acts through soil physical and chemical drivers) influences the fine root branching patterns of beech much more than soil chemical variation across soil types. To examine whether changes in root function are indeed triggered by branching events or result from root ageing and diameter growth, spatially explicit root physiological and anatomical studies across root orders are needed., (© 2024. The Author(s).)

European beech is negatively affected by climate change and a further growth decline is predicted for large parts of its distribution range. Despite the importance of this species, little is known about its genetic adaptation and especially the genetic basis of its physiological traits. Here, we used genotyping by sequencing to identify SNPs in 43 German European beech populations growing under different environmental conditions. In total, 28 of these populations were located along a precipitation and temperature gradient in northern Germany, and single tree-based hydraulic and morphological traits were available. We obtained a set of 13,493 high-quality SNPs that were used for environmental and SNP-trait association analysis. In total, 22 SNPs were identified that were significantly associated with environmental variables or specific leaf area (SLA). Several SNPs were located in genes related to stress response. The majority of the significant SNPs were located in non-coding (intergenic and intronic) regions. These may be in linkage disequilibrium with the causative coding or regulatory regions. Our study gives insights into the genetic basis of abiotic adaptation in European beech, and provides genetic resources that can be used in future studies on this species. Besides clear patterns of local adaptation to environmental conditions of the investigated populations, the analyzed morphological and hydraulic traits explained most of the explainable genetic variation. Thus, they could successfully be altered in tree breeding programs, which may help to increase the adaptation of European beech to changing environmental conditions in the future., (© 2024 The Authors. Physiologia Plantarum published by John Wiley & Sons Ltd on behalf of Scandinavian Plant Physiology Society.)

Background: The combination of radiotherapy and immunotherapy (immunoradiotherapy) has been increasingly used for treating a wide range of cancers. However, some tumors are resistant to immunoradiotherapy. We have previously shown that MER proto-oncogene tyrosine kinase (MerTK) expressed on macrophages mediates resistance to immunoradiotherapy. We therefore sought to develop therapeutics that can mitigate the negative impact of MerTK. We designed and developed a MerTK specific antisense oligonucleotide (ASO) and characterized its effects on eliciting an anti-tumor immune response in mice., Methods: 344SQR cells were injected into the right legs on day 0 and the left legs on day 4 of 8-12 weeks old female 129sv/ev mice to establish primary and secondary tumors, respectively. Radiation at a dose of 12 Gy was given to the primary tumors on days 8, 9, and 10. Mice received either anti-PD-1, anti-CTLA-4 or/and MerTK ASO starting from day 1 post tumor implantation. The composition of the tumor microenvironment and the level of MerTK on macrophages in the tumor were evaluted by flow cytometry. The expression of immune-related genes was investigated with NanoString. Lastly, the impact of MerTK ASO on the structure of the eye was histologically evaluated., Results: Remarkably, the addition of MerTK ASO to XRT+anti-PD1 and XRT+anti-CTLA4 profoundly slowed the growth of both primary and secondary tumors and significantly extended survival. The ASO significantly reduced the expression of MerTK in tumor-associated macrophages (TAMs), reprograming their phenotype from M2 to M1. In addition, MerTK ASO increased the percentage of Granzyme B + CD8 + T cells in the secondary tumors when combined with XRT+anti-CTLA4. NanoString results demonstrated that the MerTK ASO favorably modulated immune-related genes for promoting antitumor immune response in secondary tumors. Importantly, histological analysis of eye tissues demonstrated that unlike small molecules, the MerTK ASO did not produce any detectable pathology in the eyes., Conclusions: The MerTK ASO can significantly downregulate the expression of MerTK on TAMs, thereby promoting antitumor immune response. The combination of MerTK ASO with immunoradiotherapy can safely and significantly slow tumor growth and improve survival., (© 2024. The Author(s).)

The predicted increase of drought intensity in South-East Asia has raised concern about the sustainability of rubber (Hevea brasiliensis Müll. Arg.) cultivation. In order to quantify the degree of phenotypic plasticity in this important tree crop species, we analysed a set of wood and leaf traits related to the hydraulic safety and efficiency in PB260 clones from eight small-holder plantations in Jambi province, Indonesia, representing a gradient in local microclimatic and edaphic conditions. Across plots, branch embolism resistance (P50) ranged from -2.14 to -2.58 MPa. The P50 and P88 values declined, and the hydraulic safety margin increased, with an increase in the mean annual vapour pressure deficit (VPD). Among leaf traits, only the changes in specific leaf area were related to the differences in evaporative demand. These variations of hydraulic trait values were not related to soil moisture levels. We did not find a trade-off between hydraulic safety and efficiency, but vessel density (VD) emerged as a major trait associated with both safety and efficiency. The VD, and not vessel diameter, was closely related to P50 and P88 as well as to specific hydraulic conductivity, the lumen-to-sapwood area ratio and the vessel grouping index. In conclusion, our results demonstrate some degree of phenotypic plasticity in wood traits related to hydraulic safety in this tropical tree species, but this is only in response to the local changes in evaporative demand and not soil moisture. Given that VPD may increasingly limit plant growth in a warmer world, our results provide evidence of hydraulic trait changes in response to a rising evaporative demand., (© The Author(s) 2023. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

There is no currently approved adoptive cellular therapy for solid tumors. Pre-clinical and clinical studies have demonstrated that low-dose radiotherapy (LDRT) can enhance intratumoral T cell infiltration and efficacy. This case report describes a 71-year-old female patient with rectal mucosal melanoma that had developed metastases to liver, lung, mediastinum, axillary nodes, and brain. After systemic therapies had failed, she enrolled in the radiation sub-study of our phase-I clinical trial exploring the safety and efficacy of afamitresgene autoleucel (afami-cel), genetically engineered T cells with a T cell receptor (TCR) targeting the MAGE-A4 tumor antigen in patients with advanced malignancies (NCT03132922). Prior to the infusion of afami-cel, she received concurrent lymphodepleting chemotherapy and LDRT at 5.6 Gy/4 fractions to the liver. Time to partial response was 10 weeks, and duration of overall response was 18.4 weeks. Although the patient progressed at 28 weeks, the disease was well controlled after high-dose radiotherapy to liver metastases and checkpoint inhibitors. As of the last follow-up, she remains alive over two years after LDRT and afami-cel therapy. This report suggests that afami-cel in combination with LDRT safely enhanced clinical benefit. This provides evidence for further exploring the benefit of LDRT in TCR-T cell therapy., (Copyright © 2022 The Author(s). Published by Wolters Kluwer Health, Inc.)

The combination of radiation therapy (RT) and immunotherapy has emerged as a promising treatment option in oncology. Historically, x-ray radiation (XRT) has been the most commonly used form of RT. However, proton beam therapy (PBT) is gaining recognition as a viable alternative, as it has been shown to produce similar outcomes to XRT while minimizing off-target effects. The effects of PBT on the antitumor immune response have only just begun to be described, and to our knowledge no studies to date have examined the effect of PBT as part of a combinatorial immunoradiotherapeutic strategy. Here, using a 2-tumor model of lung cancer in mice, we show that PBT in tandem with an anti-PD1 antibody substantially reduced growth in both irradiated and unirradiated tumors. This was accompanied by robust activation of the immune response, as evidenced by whole-tumor and single-cell RNA sequencing showing upregulation of a multitude of immune-related transcripts. This response was further significantly enhanced by the injection of the tumor to be irradiated with NBTXR3 nanoparticles. Tumors of mice treated with the triple combination exhibited increased infiltration and activation of cytotoxic immune cells. This triple combination eradicated both tumors in 37.5% of the treated mice and showed robust long-term immunity to cancer.

Forests account for nearly 90 % of the world's terrestrial biomass in the form of carbon and they support 80 % of the global biodiversity. To understand the underlying forest dynamics, we need a long-term but also relatively high-frequency, networked monitoring system, as traditionally used in meteorology or hydrology. While there are numerous existing forest monitoring sites, particularly in temperate regions, the resulting data streams are rarely connected and do not provide information promptly, which hampers real-time assessments of forest responses to extreme climate events. The technology to build a better global forest monitoring network now exists. This white paper addresses the key structural components needed to achieve a novel meta-network. We propose to complement - rather than replace or unify - the existing heterogeneous infrastructure with standardized, quality-assured linking methods and interacting data processing centers to create an integrated forest monitoring network. These automated (research topic-dependent) linking methods in atmosphere, biosphere, and pedosphere play a key role in scaling site-specific results and processing them in a timely manner. To ensure broad participation from existing monitoring sites and to establish new sites, these linking methods must be as informative, reliable, affordable, and maintainable as possible, and should be supplemented by near real-time remote sensing data. The proposed novel meta-network will enable the detection of emergent patterns that would not be visible from isolated analyses of individual sites. In addition, the near real-time availability of data will facilitate predictions of current forest conditions (nowcasts), which are urgently needed for research and decision making in the face of rapid climate change. We call for international and interdisciplinary efforts in this direction., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Zweifel Roman reports financial support was provided by Federal Office for the Environment., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Diverse factors contribute to the limited clinical response to radiotherapy (RT) and immunotherapy in metastatic non-small cell lung cancer (NSCLC), among which is the ability of these tumors to recruit a retinue of suppressive immune cells-such as M2 tumor-associated macrophages (TAM)-thereby establishing an immunosuppressive tumor microenvironment that contributes to tumor progression and radio resistance. M2 TAMs are activated by the STAT6 signaling pathway. Therefore, we targeted STAT6 using an antisense oligonucleotide (ASO) along with hypofractionated RT (hRT; 3 fractions of 12 Gy each) to primary tumors in three bilateral murine NSCLC models (Lewis lung carcinoma, 344SQ-parental, and anti-PD-1-resistant 344SQ lung adenocarcinomas). We found that STAT6 ASO plus hRT slowed growth of both primary and abscopal tumors, decreased lung metastases, and extended survival. Interrogating the mechanism of action showed reduced M2 macrophage tumor infiltration, enhanced TH1 polarization, improved T-cell and macrophage function, and decreased TGFβ levels. The addition of anti-PD-1 further enhanced systemic antitumor responses. These results provide a preclinical rationale for the pursuit of an alternative therapeutic approach for patients with immune-resistant NSCLC., (©2023 American Association for Cancer Research.)

Increasing exposure to climate warming-related drought and heat threatens forest vitality in many regions on earth, with the trees' vulnerability likely depending on local climatic aridity, recent climate trends, edaphic conditions, and the drought acclimatization and adaptation of populations. Studies exploring tree species' vulnerability to climate change often have a local focus or model the species' entire distribution range, which hampers the separation of climatic and edaphic drivers of drought and heat vulnerability. We compared recent radial growth trends and the sensitivity of growth to drought and heat in central populations of a widespread and naturally dominant tree species in Europe, European beech (Fagus sylvatica), at 30 forest sites across a steep precipitation gradient (500-850 mm year -1 ) of short length to assess the species' adaptive potential. Size-standardized basal area increment remained more constant during the period of accelerated warming since the early 1980s in populations with >360 mm growing season precipitation (April-September), while growth trends were negative at sites with <360 mm. Climatic drought in June appeared as the most influential climatic factor affecting radial growth, with a stronger effect at drier sites. A decadal decrease in the climatic water balance of the summer was identified as the most important factor leading to growth decline, which is amplified by higher stem densities. Inter-annual growth variability has increased since the early 1980s, and variability is generally higher at drier and sandier sites. Similarly, within-population growth synchrony is higher at sandier sites and has increased with a decrease in the June climatic water balance. Our results caution against predicting the drought vulnerability of trees solely from climate projections, as soil properties emerged as an important modulating factor. We conclude that beech is facing recent growth decline at drier sites in the centre of its distribution range, driven by climate change-related climate aridification., (© 2022 The Authors. Global Change Biology published by John Wiley & Sons Ltd.)

Leaf morphological and physiological traits control the carbon and water relations of mature trees and are determinants of drought tolerance, but it is not well understood how they are modified in response to water deficits. We analysed five sun-canopy leaf traits (mean leaf size (LS), specific leaf area (SLA), Huber value (HV), water potential at turgor loss point (Ψ tlp ) and foliar carbon isotope signature (δ 13 C)) in European beech (Fagus sylvatica L.) across three precipitation gradients sampled in moist (2010), dry (2019) and very dry (2018) summers, and tested their response to short-term water deficits (climatic water balance (CWB) preceding sample collection) and long-term water availability (mean annual precipitation (MAP), plant-available soil water capacity (AWC) and neighbourhood competition). Across the 34 sites, LS varied seven-fold (3.9-27.0 cm 2 ), SLA four-fold (77.1-306.9 cm²·g -1 ) and HV six-fold (1.0-6.65 cm 2 ·m -2 ). In the 2018 dataset, LS showed a negative and HV a positive relationship to MAP, which contradicts relations found in multi-species samples. Average Ψ tlp ranged from -1.90 to -2.62 MPa and decreased across the sites with decreasing CWB in the month prior to measurement, as well as with decreasing MAP and AWC in 2019. Studied leaf traits varied considerably between years, suggesting that mast fruiting and the severe 2018 drought caused the formation of smaller leaves. We conclude that sun-canopy leaf traits of European beech exhibit considerable plasticity in response to climatic and edaphic aridity, and that osmotic adjustment may be an important element in the drought response strategy of this anisohydric tree species., (© 2021 The Authors. Plant Biology published by John Wiley & Sons Ltd on behalf of German Society for Plant Sciences, Royal Botanical Society of the Netherlands.)

Crucial for the climate adaptation of trees is a xylem anatomical structure capable of adjusting to changing water regimes. Although species comparisons across climate zones have demonstrated anatomical change in response to altered water availability and tree height, less is known about the adaptability of tree vascular systems to increasing water deficits at the intraspecific level. Information on the between-population and within-population variability of xylem traits helps assessing a species' ability to cope with climate change. We investigated the variability of wood anatomical and related hydraulic traits in terminal branches of European beech (Fagus sylvatica L.) trees across a precipitation gradient (520-890 mm year-1) and examined the influence of climatic water balance (CWB), soil water capacity (AWC), neighborhood competition (CI), tree height and branch age on these traits. Furthermore, the relationship between xylem anatomical traits and embolism resistance (P50) was tested. Within-population trait variation was larger than between-population variation. Vessel diameter, lumen-to-sapwood area ratio and potential conductivity of terminal branches decreased with decreasing CWB, but these traits were not affected by AWC, whereas vessel density increased with an AWC decrease. In contrast, none of the studied anatomical traits were influenced by variation in tree height (21-34 m) or CI. Branch age was highly variable (2-22 years) despite equal diameter and position in the flow path, suggesting different growth trajectories in the past. Vessel diameter decreased, and vessel density increased, with increasing branch age, reflecting negative annual radial growth trends. Although vessel diameter was not related to P50, vessel grouping index and lumen-to-sapwood area ratio showed a weak, though highly significant, positive relationship to P50. We conclude that the xylem anatomy of terminal tree-top branches in European beech is modified in response to increasing climatic aridity and/or decreasing soil water availability, independent of a tree height effect., (© The Author(s) 2022. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The efficacy of immunoradiotherapy consisting of radiation therapy and immune checkpoint blockade relies on effectively promoting the systemic antitumor immune response's activation while simultaneously reducing local factors favoring immune suppression. We previously demonstrated that NBTXR3, a nanoparticle radioenhancer, significantly improved immune responses in a murine anti-PD1-resistant metastatic lung cancer model. We hypothesize that radioactivated-NBTXR3 addition to anti-PD1 and a second-generation anti-CTLA4 could improve treatment effectiveness. To test this hypothesis, we inoculated mice with 344SQR cells in the right and left legs to establish primary and secondary tumors. The primary tumors were intratumorally injected with NBTXR3 nanoparticles on day 7, followed by three fractions of 12 Gy radiation on days 8, 9, and 10. The secondary tumors received two fractions of 1Gy radiation on days 13 and 14. Multiple rounds of anti-PD1, anti-CTLA4 or nonfucosylated anti-CTLA4 were given to the mice. Immune profiling of the tumors revealed that the combination of NBTXR3 with immunoradiotherapy significantly upregulated the activities of a wide range of antitumor immune pathways and reduced the abundance of regulatory suppressor T cells. This combination effectively eradicated the primary and secondary tumors and increased animal survival to 75%. Remarkably, previously treated with NBTXR3-containing treatment, the survivor mice exhibited a long-lasting antitumor memory immune response. This data provides compelling evidence of the efficacy of NBTXR3 to synergize with the immunoradiotherapy approach when combined with an anti-PD1 and multiple checkpoints such as a second generation anti-CTLA4 and show the potential for clinical uses of antitumor immunomodulatory effects of NBTXR3., Competing Interests: JWW reports research support from GlaxoSmithKline, Bristol Meyers Squibb, Merck, Nanobiotix, RefleXion Alkermes, Artidis, Mavu Pharma, Takeda, Varian, and Checkmate Pharmaceuticals. JWW serves on the scientific advisory board for Legion Healthcare Partners, RefleXion Medical, MolecularMatch, Merck, AstraZeneca, Aileron Therapeutics, OncoResponse, Checkmate Pharmaceuticals, Mavu Pharma, Alpine Immune Sciences, Ventana Medical Systems, Nanobiotix, China Medical Tribune, GI Innovation, Genentech and Nanorobotics. JWW is on Speaking Engagements for Ventana Medical Systems, US Oncology, Alkermes, and Boehringer Ingelheim. He is co-founder of Healios, MolecularMatch, OncoResponse and serves as an advisor to Astra Zeneca, OncoResponse, Merck, MolecularMatch, Incyte, Aileron and Nanobiotix. JWW holds stock or ownership in Alpine Immune Sciences, Checkmate Pharmaceuticals, Healios, Mavu Pharma, Legion Healthcare Partners, MolecularMatch, Nanorobotics, OncoResponse, and RefleXion. JWW has accepted honoraria in the form of travel costs from Nanobiotix, RefleXion, Varian, Shandong University, The Korea Society of Radiology, Aileron Therapeutics and Ventana. JWW has the following patents; MP470 amuvatinib, MRX34 regulation of PDL1, XRT technique to overcome immune resistance. MD Anderson Cancer Center has a trademark for XRTTM. SP and JS are employees of Nanobiotix. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. This study received funding from Nanobiotix. The funder had the following involvement with the study: experiment design, data analysis, and manuscript writing., (Copyright © 2022 Hu, Paris, Bertolet, Barsoumian, Wang, Da Silva, Patel, Nguyen, Doss, Huang, Hsu, Leyton, Voss, Masrorpour, Leuschner, Pietz, Puebla-Osorio, Gandhi, Nguyen, Wang, Cortez and Welsh.)

Background: While improvements in immunoradiotherapy have significantly improved outcomes for cancer patients, this treatment approach has nevertheless proven ineffective at controlling the majority of malignancies. One of the mechanisms of resistance to immunoradiotherapy is that immune cells may be suppressed via the myriad of different immune checkpoint receptors. Therefore, simultaneous blockade of multiple immune checkpoint receptors may enhance the treatment efficacy of immunoradiotherapy., Methods: We combined NBTXR3-enhanced localized radiation with the simultaneous blockade of three different checkpoint receptors: PD1, LAG3, and TIGIT, and tested the treatment efficacy in an anti-PD1-resistant lung cancer model in mice. 129 Sv/Ev mice were inoculated with fifty thousand αPD1-resistant 344SQR cells in the right leg on day 0 to establish primary tumors and with the same number of cells in the left leg on day 4 to establish the secondary tumors. NBTXR3 was intratumorally injected into the primary tumors on day 7, which were irradiated with 12 Gy on days 8, 9, and 10. Anti-PD1 (200 µg), αLAG3 (200 µg), and αTIGIT (200 µg) were given to mice by intraperitoneal injections on days 5, 8, 11, 14, 21, 28, 35, and 42., Results: This nanoparticle-mediated combination therapy is effective at controlling the growth of irradiated and distant unirradiated tumors, enhancing animal survival, and is the only one that led to the destruction of both tumors in approximately 30% of the treated mice. Corresponding with this improved response is robust activation of the immune response, as manifested by increased numbers of immune cells along with a transcriptional signature of both innate and adaptive immunity within the tumor. Furthermore, mice treated with this combinatorial therapy display immunological memory response when rechallenged by the same cancer cells, preventing tumor engraftment., Conclusion: Our results strongly attest to the efficacy and validity of combining nanoparticle-enhanced radiotherapy and simultaneous blockade of multiple immune checkpoint receptors and provide a pre-clinical rationale for investigating its translation into human patients., (© 2022. The Author(s).)

The growth of past, present, and future forests was, is and will be affected by climate variability. This multifaceted relationship has been assessed in several regional studies, but spatially resolved, large-scale analyses are largely missing so far. Here we estimate recent changes in growth of 5800 beech trees (Fagus sylvatica L.) from 324 sites, representing the full geographic and climatic range of species. Future growth trends were predicted considering state-of-the-art climate scenarios. The validated models indicate growth declines across large region of the distribution in recent decades, and project severe future growth declines ranging from -20% to more than -50% by 2090, depending on the region and climate change scenario (i.e. CMIP6 SSP1-2.6 and SSP5-8.5). Forecasted forest productivity losses are most striking towards the southern distribution limit of Fagus sylvatica, in regions where persisting atmospheric high-pressure systems are expected to increase drought severity. The projected 21 st century growth changes across Europe indicate serious ecological and economic consequences that require immediate forest adaptation., (© 2022. The Author(s).)

Xylem embolism resistance has been identified as a key trait with a causal relation to drought-induced tree mortality, but not much is known about its intra-specific trait variability (ITV) in dependence on environmental variation. We measured xylem safety and efficiency in 300 European beech (Fagus sylvatica L.) trees across 30 sites in Central Europe, covering a precipitation reduction from 886 to 522 mm year -1 . A broad range of variables that might affect embolism resistance in mature trees, including climatic and soil water availability, competition, and branch age, were examined. The average P 50 value varied by up to 1 MPa between sites. Neither climatic aridity nor structural variables had a significant influence on P 50 . However, P 50 was less negative for trees with a higher soil water storage capacity, and positively related to branch age, while specific conductivity (K s ) was not significantly associated with either of these variables. The greatest part of the ITV for xylem safety and efficiency was attributed to random variability within populations. We conclude that the influence of site water availability on P 50 and K s is low in European beech, and that the high degree of within-population variability for P 50 , partly due to variation in branch age, hampers the identification of a clear environmental signal., (© 2022. The Author(s).)

The iso/anisohydric continuum has been used to classify tree species' drought response strategies. The range over which stomata are regulating leaf water potential (ψl) before turgor loss occurs can be described with metrics such as the dependence of ψl on soil water potential (ψsoil) and the size of 'hydroscape area' (HA), but corresponding field data from adult trees are scarce. We examined the stomatal conductance (gs)-ψl relationship in its temporal (diurnal vs seasonal and interannual) and spatial (within-crown vs between-site) variation in European beech, using extensive ψl and gs measurements in the canopy of four beech stands across a precipitation gradient, and complemented the data set by published ψl and gs measurements in further Central European beech stands (including the extreme 2018 drought) in order to cover the full water potential operation space of the species. Both metrics characterize beech as a strictly anisohydric species with δψl/δψsoil >> 1 and HA = 4 MPa2. However, stomates close sensitively in response to increasing vapor pressure deficit, disproving the widely assumed dependence of large ψl variation on looser stomatal control. Characterizing the water status regulation mechanisms of trees requires separating diurnal from day-to-day variation in ψl and gs. The large diurnal and seasonal ψl variation in beech leaves is partly caused by a low leaf tissue elasticity, suggesting that a whole-plant perspective with consideration of osmotic and elastic tissue properties and stem and root hydraulics is needed for fully understanding ψl regulation and the drought tolerance strategy of trees., (© The Author(s) 2021. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permission@oup.com.)

Biodiversity and ecosystem functions are highly threatened by global change. It has been proposed that geodiversity can be used as an easy-to-measure surrogate of biodiversity to guide conservation management. However, so far, there is mixed evidence to what extent geodiversity can predict biodiversity and ecosystem functions at the regional scale relevant for conservation planning. Here, we analyse how geodiversity computed as a compound index is suited to predict the diversity of four taxa and associated ecosystem functions in a tropical mountain hotspot of biodiversity and compare the results with the predictive power of environmental conditions and resources (climate, habitat, soil). We show that combinations of these environmental variables better explain species diversity and ecosystem functions than a geodiversity index and identified climate variables as more important predictors than habitat and soil variables, although the best predictors differ between taxa and functions. We conclude that a compound geodiversity index cannot be used as a single surrogate predictor for species diversity and ecosystem functions in tropical mountain rain forest ecosystems and is thus little suited to facilitate conservation management at the regional scale. Instead, both the selection and the combination of environmental variables are essential to guide conservation efforts to safeguard biodiversity and ecosystem functions., (© 2021. The Author(s).)

Despite multiple therapeutic approaches, the presence of liver metastases carries a guarded prognosis, urgently necessitating further clinical and scientific research to develop curative interventions. The liver is an immunoprivileged organ that suppresses the effectiveness of immunotherapies in patients with hepatic metastases. Cancer immunotherapies have been successfully bolstered by low-dose radiotherapy (LDRT), which is capable of reprogramming the tumor microenvironment (TME) from an immunosuppressive to an immunostimulatory one. Likewise, LDRT may be able to revoke the immune privilege enjoyed by the liver, permitting successful immunotherapies there. Here, we first review challenges that face the treatment of liver metastases. We next outline emerging preclinical and clinical evidence supporting enhanced systemic tumor control of LDRT in the context of cancer immunotherapy. Finally, we will discuss the rationale of combining liver-directed LDRT with immunostimulatory strategies to overcome immune resistance and achieve better clinical response. This notion is supported by a recent case study in which a patient who had progressed following T cell therapy experienced a complete response after LDRT to the liver., Competing Interests: JW reports grants from Varian Medical Systems, Bristol-Meyers Squibb; personal fees and other from Alpine Immune Sciences, Legion Healthcare Partners, Molecular Match, Nanorobotix, OncoResponse, and RefleXion Medical; grants and personal fees from Nanobiotics; grants, personal fees or equity from Checkmate Pharmaceuticals. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 He, Barsoumian, Bertolet, Verma, Leuschner, Koay, Ludmir, Hsu, Pisipati, Voss, Puebla-Osorio, Cortez and Welsh.)

Many experiments have shown that biodiversity enhances ecosystem functioning. However, we have little understanding of how environmental heterogeneity shapes the effect of diversity on ecosystem functioning and to what extent this diversity effect is mediated by variation in species richness or species turnover. This knowledge is crucial to scaling up the results of experiments from local to regional scales. Here we quantify the diversity effect and its components-that is, the contributions of variation in species richness and species turnover-for 22 ecosystem functions of microorganisms, plants and animals across 13 major ecosystem types on Mt Kilimanjaro, Tanzania. Environmental heterogeneity across ecosystem types on average increased the diversity effect from explaining 49% to 72% of the variation in ecosystem functions. In contrast to our expectation, the diversity effect was more strongly mediated by variation in species richness than by species turnover. Our findings reveal that environmental heterogeneity strengthens the relationship between biodiversity and ecosystem functioning and that species richness is a stronger driver of ecosystem functioning than species turnover. Based on a broad range of taxa and ecosystem functions in a non-experimental system, these results are in line with predictions from biodiversity experiments and emphasize that conserving biodiversity is essential for maintaining ecosystem functioning., (© 2021. The Author(s), under exclusive licence to Springer Nature Limited.)

The study of animal-habitat interactions is of primary importance for the formulation of conservation recommendations. Flying, gliding, and climbing animals have the ability to exploit their habitat in a three-dimensional way, and the vertical canopy structure in forests plays an essential role for habitat suitability. Forest bats as flying mammals may seasonally shift their microhabitat use due to differing energy demands or changing prey availability, but the patterns are not well understood. We investigated three-dimensional and seasonal habitat use by insectivorous bats in a temperate lowland old-growth forest, the Belovezhskaya Pushcha in Belarus. We acoustically sampled broadleaved and mixed coniferous plots in the forest interior and in gaps in three heights during two reproductive periods (pregnancy/lactation vs. postlactation). In canopy gaps, vertical stratification in bat activity was less pronounced than in the forest interior. Vertical activity patterns differed among species. The upper canopy levels were important foraging habitats for the open-space forager guild and for some edge-space foragers like the Barbastelle bat Barbastella barbastellus and the soprano pipistrelle Pipistrellus pygmaeus . Myotis species had highest activity levels near the ground in forest gaps. Moreover, we found species-dependent seasonal microhabitat shifts. Generally, all species and species groups considered except Myotis species showed higher activity levels during postlactation. Myotis species tended toward higher activity in the forest interior during postlactation. P .  pygmaeus switched from high activity levels in the upper canopy during pregnancy and lactation to high activity levels near the ground during postlactation. We conclude that a full comprehension of forest bat habitat use is only possible when height in canopy and seasonal patterns are considered., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2021 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Plant hydraulic traits are key for understanding and predicting tree drought responses. Information about the degree of the traits' intra-specific variability may guide the selection of drought-resistant genotypes and is crucial for trait-based modelling approaches. For the three temperate minor broadleaf tree species Acer platanoides, Carpinus betulus and Tilia cordata, we measured xylem embolism resistance (P 50 ), leaf turgor loss point (P TLP ), specific hydraulic conductivity (K S ), Huber values (HVs), and hydraulic safety margins in adult trees across a precipitation gradient. We further quantified trait variability on different organizational levels (inter-specific to within-canopy variation), and analysed its relationship to climatic and soil water availability. Although we observed a certain intra-specific trait variability (ITV) in safety-related traits (P 50 , P TLP ) with higher within-tree and between-tree than between populations variability, the magnitude was small compared to inter-specific differences, which explained 78.4% and 58.3% of the variance in P 50 and P TLP , respectively. In contrast, efficiency-related traits (K S , HV) showed a high ITV both within populations and within the crowns of single trees. Surprisingly, the observed ITV of all traits was neither driven by climatic nor soil water availability. In conclusion, the high degree of conservatism in safety-related traits highlights their potential for trait-based modelling approaches., (© 2021 The Authors New Phytologist © 2021 New Phytologist Foundation.)

Screening species-rich communities for the variation in functional traits along environmental gradients may help understanding the abiotic drivers of plant performance in a mechanistic way. We investigated tree leaf trait variation along an elevation gradient (1000-3000 m) in highly diverse neotropical montane forests to test the hypothesis that elevational trait change reflects a trend toward more conservative resource use strategies at higher elevations, with interspecific trait variation decreasing and trait integration increasing due to environmental filtering. Analysis of trait variance partitioning across the 52 tree species revealed for most traits a dominant influence of phylogeny, except for SLA, leaf thickness and foliar Ca, where elevation was most influential. The community-level means of SLA, foliar N and Ca, and foliar N/P ratio decreased with elevation, while leaf thickness and toughness increased. The contribution of intraspecific variation was substantial at the community level in most traits, yet smaller than the interspecific component. Both within-species and between-species trait variation did not change systematically with elevation. High phylogenetic diversity, together with small-scale edaphic heterogeneity, cause large interspecific leaf trait variation in these hyper-diverse Andean forests. Trait network analysis revealed increasing leaf trait integration with elevation, suggesting stronger environmental filtering at colder and nutrient-poorer sites.

We investigated the variation in tree fine root traits and their functional diversity along a local topographic gradient in a Neotropical montane forest to test if fine root trait variation along the gradient is consistent with the predictions of the root economics spectrum on a shift from acquisitive to conservative traits with decreasing resource supply. We measured five fine root functional traits in 179 randomly selected tree individuals of 100 species and analysed the variation of single traits (using Bayesian phylogenetic multilevel models) and of functional trait diversity with small-scale topography. Fine roots exhibited more conservative traits (thicker diameters, lower specific root length and nitrogen concentration) at upper slope compared with lower slope positions, but the largest proportion of variation (40-80%) was explained by species identity and phylogeny. Fine root functional diversity decreased towards the upper slopes. Our results suggest that local topography and the related soil fertility and moisture gradients cause considerable small-scale variation in fine root traits and functional diversity along tropical mountain slopes, with conservative root traits and greater trait convergence being associated with less favourable soil conditions due to environmental filtering. We provide evidence of a high degree of phylogenetic conservation in fine root traits., (© 2020 The Authors New Phytologist © 2020 New Phytologist Foundation.)

Tropical mountain ecosystems are threatened by climate and land-use changes. Their diversity and complexity make projections how they respond to environmental changes challenging. A suitable way are trait-based approaches, by distinguishing between response traits that determine the resistance of species to environmental changes and effect traits that are relevant for species' interactions, biotic processes, and ecosystem functions. The combination of those approaches with land surface models (LSM) linking the functional community composition to ecosystem functions provides new ways to project the response of ecosystems to environmental changes. With the interdisciplinary project RESPECT, we propose a research framework that uses a trait-based response-effect-framework (REF) to quantify relationships between abiotic conditions, the diversity of functional traits in communities, and associated biotic processes, informing a biodiversity-LSM. We apply the framework to a megadiverse tropical mountain forest. We use a plot design along an elevation and a land-use gradient to collect data on abiotic drivers, functional traits, and biotic processes. We integrate these data to build the biodiversity-LSM and illustrate how to test the model. REF results show that aboveground biomass production is not directly related to changing climatic conditions, but indirectly through associated changes in functional traits. Herbivory is directly related to changing abiotic conditions. The biodiversity-LSM informed by local functional trait and soil data improved the simulation of biomass production substantially. We conclude that local data, also derived from previous projects (platform Ecuador), are key elements of the research framework. We specify essential datasets to apply this framework to other mountain ecosystems.

Tropical forests represent the largest store of terrestrial biomass carbon (C) on earth and contribute over-proportionally to global terrestrial net primary productivity (NPP). How climate change is affecting NPP and C allocation to tree components in forests is not well understood. This is true for tropical forests, but particularly for African tropical forests. Studying forest ecosystems along elevation and related temperature and moisture gradients is one possible approach to address this question. However, the inclusion of belowground productivity data in such studies is scarce. On Mt. Kilimanjaro (Tanzania), we studied aboveground (wood increment, litter fall) and belowground (fine and coarse root) NPP along three elevation transects (c. 1800-3900 m a.s.l.) across four tropical montane forest types to derive C allocation to the major tree components. Total NPP declined continuously with elevation from 8.5 to 2.8 Mg C ha -1  year -1 due to significant decline in aboveground NPP, while fine root productivity (sequential coring approach) remained unvaried with around 2 Mg C ha -1  year -1 , indicating a marked shift in C allocation to belowground components with elevation. The C and N fluxes to the soil via root litter were far more important than leaf litter inputs in the subalpine Erica forest. Thus, the shift of C allocation to belowground organs with elevation at Mt. Kilimanjaro and other tropical forests suggests increasing nitrogen limitation of aboveground tree growth at higher elevations. Our results show that studying fine root productivity is crucial to understand climate effects on the carbon cycle in tropical forests.

Objective: This randomized open-label phase II study evaluated the safety and clinical activity of EP-100 plus weekly paclitaxel in patients with recurrent ovarian cancer expressing positive LHRH receptor., Methods: In a limited "run-in" dose escalation phase for EP-100, six patients were treated with ascending dose levels (13 mg/m 2 , 20 mg/m 2 , 30 mg/m 2 ). In the randomized phase, patients received weekly paclitaxel (80 mg/m 2 intravenously) plus twice weekly EP-100 (30 mg/m 2 intravenously; combination arm) or weekly paclitaxel alone (80 mg/m 2 intravenously; paclitaxel arm). The primary study endpoint was overall response rate (ORR)., Results: Forty-four patients were then randomized to either the experimental combination arm (n = 23) or the standard of care paclitaxel monotherapy arm (n = 21). The ORR was 35% (95%CI 16%-57%) for the combination arm and 33% (95% CI 15%-57%) for the paclitaxel arm. An interesting observation from an unplanned analysis was that a subset of patients with target liver lesions showed a greater overall response rate to the combination (69%) compared to paclitaxel alone (16%). The frequency of treatment-related grade 3-4 adverse events was similar between treatment arms: 48% vs 43% for the combination and paclitaxel arms, respectively., Conclusions: ORR in the EP-100 combination arm was similar to that in the group treated with paclitaxel alone; however, a subset of patients with liver metastases appeared to benefit from the combination. The addition of EP-100 did not appear to augment the adverse event profile of paclitaxel and was well tolerated., Competing Interests: Declaration of Competing Interest, (Copyright © 2020 Elsevier Inc. All rights reserved.)

Here, we examined the role of EP-100 [luteinizing hormone-releasing hormone (LHRH) ligand joined to a lytic peptide], improving the efficacy of immune checkpoint blockade. LHRH-R-positive murine ovarian cancer cells (ID8, IG10, IF5, and 2C12) were sensitive to EP-100 and were specifically killed at low micromolar levels through LHRH-R. EP-100 increased PD-L1 levels on murine ovarian cancer cells. In vivo syngeneic mouse models (ID8 and IG10) demonstrated that single-agent EP-100 reduced tumor volume, tumor weight, and ascites volume. The greatest reductions in tumor and ascites volume were observed with the combination of EP-100 with an anti-PD-L1 antibody. Immune profiling analysis showed that the population of CD8 + T cells, natural killer cells, dendritic cells, and macrophages were significantly increased in tumor and ascitic fluid samples treated with anti-PD-L1, EP-100, and the combination. However, monocytic myeloid suppressor cells, B cells, and regulatory T cells were decreased in tumors treated with anti-PD-L1, EP-100, or the combination. In vitro cytokine arrays revealed that EP-100 induced IL1α, IL33, CCL20, VEGF, and Low-density lipoprotein receptor (LDLR) secretion. Of these, we validated increasing IL33 levels following EP-100 treatment in vitro and in vivo ; we determined the specific biological role of CD8 + T-cell activation with IL33 gene silencing using siRNA and Cas9-CRISPR approaches. In addition, we found that CD8 + T cells expressed very low level of LHRH-R and were not affected by EP-100. Taken together, EP-100 treatment had a substantial antitumor efficacy, particularly in combination with an anti-PD-L1 antibody. These results warrant further clinical development of this combination., (©2020 American Association for Cancer Research.)

The authors of the above-mentioned publication noticed that the database for the beta diversity calculation was not correctly chosen.

Root exudation is a key plant function with a large influence on soil organic matter dynamics and plant-soil feedbacks in forest ecosystems. Yet despite its importance, the main ecological drivers of root exudation in mature forest trees remain to be identified. During two growing seasons, we analyzed the dependence of in situ collected root exudates on root morphology, soil chemistry and nutrient availability in six mature European beech (Fagus sylvatica L.) forests on a broad range of bedrock types. Root morphology was a major driver of root exudation across the nutrient availability gradient. A doubling of specific root length exponentially increased exudation rates of mature trees by c. 5-fold. Root exudation was also closely negatively related to soil pH and nitrogen (N) availability. At acidic and N-poor sites, where fungal biomass was reduced, exudation rates were c. 3-fold higher than at N- and base-richer sites and correlated negatively with the activity of enzymes degrading less bioavailable carbon (C) and N in the bulk soil. We conclude that root exudation increases on highly acidic, N-poor soils, in which fungal activity is reduced and a greater portion of the assimilated plant C is shifted to the external ecosystem C cycle., (© 2020 The Authors. New Phytologist © 2020 New Phytologist Trust.)

Land-use transitions can enhance the livelihoods of smallholder farmers but potential economic-ecological trade-offs remain poorly understood. Here, we present an interdisciplinary study of the environmental, social and economic consequences of land-use transitions in a tropical smallholder landscape on Sumatra, Indonesia. We find widespread biodiversity-profit trade-offs resulting from land-use transitions from forest and agroforestry systems to rubber and oil palm monocultures, for 26,894 aboveground and belowground species and whole-ecosystem multidiversity. Despite variation between ecosystem functions, profit gains come at the expense of ecosystem multifunctionality, indicating far-reaching ecosystem deterioration. We identify landscape compositions that can mitigate trade-offs under optimal land-use allocation but also show that intensive monocultures always lead to higher profits. These findings suggest that, to reduce losses in biodiversity and ecosystem functioning, changes in economic incentive structures through well-designed policies are urgently needed.

Fine roots (≤2 mm) consume a large proportion of photosynthates and thus play a key role in the global carbon cycle, but our knowledge about fine root biomass, production, and turnover across environmental gradients is insufficient, especially in tropical ecosystems. Root system studies along elevation transects can produce valuable insights into root trait-environment relationships and may help to explore the evidence for a root economics spectrum (RES) that should represent a trait syndrome with a trade-off between resource acquisitive and conservative root traits. We studied fine root biomass, necromass, production, and mean fine root lifespan (the inverse of fine root turnover) of woody plants in six natural tropical ecosystems (savanna, four tropical mountain forest types, tropical alpine heathland) on the southern slope of Mt. Kilimanjaro (Tanzania) between 900 and 4,500 m a.s.l. Fine root biomass and necromass showed a unimodal pattern along the slope with a peak in the moist upper montane forest (~2,800 m), while fine root production varied little between savanna and upper montane forest to decrease toward the alpine zone. Root:shoot ratio (fine root biomass and production related to aboveground biomass) in the tropical montane forest increased exponentially with elevation, while it decreased with precipitation and soil nitrogen availability (decreasing soil C:N ratio). Mean fine root lifespan was lowest in the ecosystems with pronounced resource limitation (savanna at low elevation, alpine heathland at high elevation) and higher in the moist and cool forest belt (~1,800-3,700 m). The variation in root traits across the elevation gradient fits better with the concept of a multi-dimensional RES, as root tissue density and specific root length showed variable relations to each other, which does not agree with a simple trade-off between acquisitive and conservative root traits. In conclusion, despite large variation in fine root biomass, production, and morphology among the different plant species and ecosystems, a general belowground shift in carbohydrate partitioning is evident from 900 to 4,500 m a.s.l., suggesting that plant growth is increasingly limited by nutrient (probably N) shortage toward higher elevations., (Copyright © 2020 Sierra Cornejo, Hertel, Becker, Hemp and Leuschner.)

Agricultural intensification has led to drastic population declines in Europe's arable plant vegetation, and continuous monitoring is a prerequisite for assessing measures to increase and conserve remnant populations of endangered arable plant species. Unfortunately, strong variation in plot sizes and in-field locations makes comparison of current arable plant monitoring approaches difficult. This study compares different relevé approaches in conventionally managed arable fields in Northwest German farmland with respect to plant species detection success and time expenditure. We compared species detection rate and expenditure of time of six different relevé types in 45 conventionally managed arable fields (each 15 fields of wheat, maize, and rapeseed): field "Interior" plots (50 × 2 m); field edge plots: "Edge&#95;30" (30 × 2 m), "Edge&#95;50" (50 × 2 m), and "Edge&#95;500" (500 × 1 m); "Subplots" (four dispersed plots of 5 × 1 m); and "Corner" plots (50 × 2 m). To determine species detection rate, the species richness recorded with a survey method was related to the field's total plant species number as estimated from a survey of the entire field edge zone. With a species detection rate of 8.3% (median), interior plots were inadequate for characterizing the field's arable plant vegetation. Edge&#95;500 plots yielded the highest proportion of the field's arable plant species pool (75.6%, including taxa of conservation value), followed by "Corner" plots (45.8%) and "Sublots" (32.6%). Edge&#95;50 and Edge&#95;30 plots detected less than 25% of the field's species pool. The average time needed for a relevé was 20 min in Edge500 plots and 5-11 min in the other plot types. We suggest implementing Edge&#95;500 plots as a standard monitoring approach in conventionally managed farmland due to its favorable ratio of detection success to expenditure of time. Our findings should be compared to methodological studies conducted in other regions, in different farmland management systems, and in landscapes of variable complexities.

With the prospect of climate change and more frequent El Niño-related dry spells, the drought tolerance of oil palm (Elaeis guineensis Jacq.), one of the most important tropical crop species, is of major concern. We studied the influence of soil water availability and palm height on the plasticity of xylem anatomy of oil palm fronds and their embolism resistance at well-drained and seasonally flooded riparian sites in lowland Sumatra, Indonesia. We found overall mean P12 and P50 values, i.e., the xylem pressures at 12% or 50% loss of hydraulic conductance, of -1.05 and - 1.86 MPa, respectively, indicating a rather vulnerable frond xylem of oil palm. This matches diurnal courses of stomatal conductance, which in combination with the observed low xylem safety evidence a sensitive water loss regulation. While the xylem anatomical traits vessel diameter (Dh), vessel density and potential hydraulic conductivity (Kp) were not different between the sites, palms in the moister riparian plots had on average by 0.4 MPa higher P50 values than plants in the well-drained plots. This could largely be attributed to differences in palm height between systems. As a consequence, palms of equal height had 1.3 MPa less negative P50 values in the moister riparian plots than in the well-drained plots. While palm height was positively related to P50, Dh and Kp decreased with height. The high plasticity in embolism resistance may be an element of the drought response strategy of oil palm, which, as a monocot, has a relatively deterministic hydraulic architecture. We conclude that oil palm fronds develop a vulnerable water transport system, which may expose the palms to increasing drought stress in a warmer and drier climate. However, the risk of hydraulic failure may be reduced by considerable plasticity in the hydraulic system and the environmental control of embolism resistance, and a presumably large stem capacitance., (© The Author(s) 2019. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

EP-100 is a synthetic lytic peptide that specifically targets the gonadotropin-releasing hormone receptor on cancer cells. To extend the utility of EP-100, we aimed to identify effective combination therapies with EP-100 for ovarian cancer and explore potential mechanisms of this combination. A series of in vitro (MTT assay, immunoblot analysis, reverse-phase protein array, comet assay, and immunofluorescence staining) and in vivo experiments were carried out to determine the biological effects of EP-100 alone and in combination with standard-of-care drugs. EP-100 decreased the viability of ovarian cancer cells and reduced tumor growth in orthotopic mouse models. Of five standard drugs tested (cisplatin, paclitaxel, doxorubicin, topotecan, and olaparib), we found that the combination of EP-100 and olaparib was synergistic in ovarian cancer cell lines. Further experiments revealed that combined treatment of EP-100 and olaparib significantly increased the number of nuclear foci of phosphorylated histone H2AX. In addition, the extent of DNA damage was significantly increased after treatment with EP-100 and olaparib in comet assay. We performed reverse-phase protein array analyses and identified that the PI3K/AKT pathway was inhibited by EP-100, which we validated with in vitro experiments. In vivo experiment using the HeyA8 mouse model demonstrated that mice treated with EP-100 and olaparib had lower tumor weights (0.06 ± 0.13 g) than those treated with a vehicle (1.19 ± 1.09 g), EP-100 alone (0.62 ± 0.78 g), or olaparib alone (0.50 ± 0.63 g). Our findings indicate that combining EP-100 with olaparib is a promising therapeutic strategy for ovarian cancer., (©2019 American Association for Cancer Research.)

In high-elevation grasslands, plants can encounter periods with high air temperature while the soil remains cold, which may lead to a temporary mismatch in the physiological activity of leaves and roots. In a climate chamber experiment with graminoid species from three elevations (4400, 2400, and 250 m a.s.l.), we tested the hypothesis that soil temperature can influence photosynthesis and stomatal conductance independently of air temperature. Soil monoliths with swards of Kobresia pygmaea (high alpine), Nardus stricta (lower alpine), and Deschampsia flexuosa (upper lowland) were exposed to soil temperatures of 25, 15, 5, and -2°C and air temperatures of 20 and 10°C for examining the effect of independent soil and air temperature variation on photosynthesis, leaf dark respiration, and stomatal conductance and transpiration. Soil frost (-2°C) had a strong negative effect on gas exchange and stomatal conductance in all three species, independent of the elevation of origin. Leaf dark respiration was stimulated by soil frost in D. flexuosa , but not in K. pygmaea , which also had a lower temperature optimum of photosynthesis. Soil cooling from 15 to 5°C did not significantly reduce stomatal conductance and gas exchange in any of the species. We conclude that all three graminoids are able to maintain a relatively high root water uptake in cold, non-frozen soil, but the high-alpine K. pygmaea seems to be especially well adapted to warm shoot - cold root episodes, as it has a higher photosynthetic activity at 10 than 20°C air temperature and does not up-regulate leaf dark respiration upon soil freezing, as was observed in the grasses from warmer climates.

In forest ecosystems, ectomycorrhizal (ECM) fungi are important for plant growth and soil biogeochemical processes. The biochemical composition of ECM mycelium is an important fungal effect trait with consequences for its decomposition rate, and consequently on soil carbon pools and plant nutrition. Although the link between ECM fungi and leaf litter-released nutrients is well known, the response of ECM fungal biochemical composition to different leaf litter species remains poorly understood. To determine how leaf litter quality influences ECM fungi's biochemical profiles, we planted young beech trees in an oak forest and replaced the natural leaf litter with that of European beech (Fagus sylvatica), ash (Fraxinus excelsior), maple (Acer pseudoplatanus), or lime (Tilia cordata). We assessed the biochemical profiles of ECM root tips colonized by common fungal taxa in temperate forests (i.e., Cenococcum geophilum, Inocybe sp., and Lactarius subdulcis), using attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR). ECM fungal biochemical composition changed with leaf litter species. Changes were apparent in the infrared absorption bands assigned to functional groups of lipids, amides, and carbohydrates. C. geophilum and L. subdulcis exhibited large spectral differences corresponding to the initial pattern of leaf litter chemical composition between samples collected in the beech and ash leaf litter treatments. In contrast, Inocybe sp. was influenced by lime, but with no differences between samples from ash or beech leaf litter treatments. Although the spectral bands affected by leaf litter type differed among ECM fungi, they were mainly related to amides, indicating a dynamic response of the fungal proteome to soil nutritional changes. Overall, the results indicate that the biochemical response of ECM fungi to leaf litter species varies among ECM fungal species and suggests that the biochemical composition of ECM mycelium is a fungal response trait, sensitive to environmental changes such as shifts in leaf litter species.

The branch xylem of six important Inner Asian southern boreal forest trees was studied for wood-anatomical and hydraulic traits in order to infer the species' drought tolerance from embolism resistance, potential hydraulic conductivity, mean conduit diameters and conduit density. The only studied angiosperm tree, Betula pendula Roth, was much more sensitive to cavitation than all five conifers (evergreen or summer-green), even when using 88% loss of conductivity (P88) in birch, but 50% (P50) in the conifers as critical thresholds. This suggests that pioneer birch forests, which have widely replaced the conifer climax forests after anthropogenic disturbance (e.g., logging, man-made fire), are more vulnerable to climate warming-induced drought than the original conifer forests. In contrast to expectation, the generally more drought-exposed light taiga species (Larix sibirica Ledeb., Pinus sylvestris L.) did not have consistently lower P50 and P88 values than the dark taiga conifers, suggesting that other drought survival traits are equally important. Among the dark-taiga species, only Pinus sibirica Du Tour, but not Abies sibirica Ledeb. and Picea obovata Ledeb., had relatively high P50 values indicating higher vulnerability. In the light-taiga forest, P. sylvestris revealed lower embolism resistance than L. sibirica. In the face of rapid climate warming and drying in Inner Asia, the drought survival strategies of southern boreal tree species deserve further intensive study, which should include other drought survival traits., (© The Author(s) 2018. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

With 450,000 km 2 Kobresia (syn. Carex) pygmaea dominated pastures in the eastern Tibetan highlands are the world's largest pastoral alpine ecosystem forming a durable turf cover at 3000-6000 m a.s.l. Kobresia's resilience and competitiveness is based on dwarf habit, predominantly below-ground allocation of photo assimilates, mixture of seed production and clonal growth, and high genetic diversity. Kobresia growth is co-limited by livestock-mediated nutrient withdrawal and, in the drier parts of the plateau, low rainfall during the short and cold growing season. Overstocking has caused pasture degradation and soil deterioration over most parts of the Tibetan highlands and is the basis for this man-made ecosystem. Natural autocyclic processes of turf destruction and soil erosion are initiated through polygonal turf cover cracking, and accelerated by soil-dwelling endemic small mammals in the absence of predators. The major consequences of vegetation cover deterioration include the release of large amounts of C, earlier diurnal formation of clouds, and decreased surface temperatures. These effects decrease the recovery potential of Kobresia pastures and make them more vulnerable to anthropogenic pressure and climate change. Traditional migratory rangeland management was sustainable over millennia, and possibly still offers the best strategy to conserve and possibly increase C stocks in the Kobresia turf., (Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Old-growth and primeval forests are passing through a natural development cycle with recurring stages of forest development. Several methods for assigning patches of different structure and size to forest development stages or phases do exist. All currently existing classification methods have in common that a priori assumptions about the characteristics of certain stand structural attributes such as deadwood amount are made. We tested the hypothesis that multivariate datasets of primeval beech forest stand structure possess an inherent, aggregated configuration of data points with individual clusters representing forest development stages. From two completely mapped primeval beech forests in Albania, seven ecologically important stand structural attributes characterizing stand density, regeneration, stem diameter variation and amount of deadwood are derived at 8216 and 9666 virtual sampling points (moving window, focal filtering). K-means clustering is used to detect clusters in the datasets (number of clusters (k) between 2 and 5). The quality of the single clustering solutions is analyzed with average silhouette width as a measure for clustering quality. In a sensitivity analysis, clustering is done with datasets of four different spatial scales of observation (200, 500, 1000 and 1500 m 2 , circular virtual plot area around sampling points) and with two different kernels (equal weighting of all objects within a plot vs. weighting by distance to the virtual plot center)., Results: The clustering solutions succeeded in detecting and mapping areas with homogeneous stand structure. The areas had extensions of more than 200 m 2 , but differences between clusters were very small with average silhouette widths of less than 0.28. The obtained datasets had a homogeneous configuration with only very weak trends for clustering., Conclusions: Our results imply that forest development takes place on a continuous scale and that discrimination between development stages in primeval beech forests is splitting continuous datasets at selected thresholds. For the analysis of the forest development cycle, direct quantification of relevant structural features or processes might be more appropriate than classification. If, however, the study design demands classification, our results can justify the application of conventional forest development stage classification schemes rather than clustering.

Climate extremes are predicted to become more frequent and intense in future. Thus, understanding how trees respond to adverse climatic conditions is crucial for evaluating possible future changes in forest ecosystem functioning. Although much information about climate effects on the growth of temperate trees has been collected in recent decades, our understanding of the influence of forest management legacies on climate-growth relationships is still limited. We used individual tree-ring chronologies from managed and unmanaged European beech forests, located in the same growth district (i.e. with almost identical climatic and soil conditions), to examine how forest management legacies (recently managed with selection cutting, >20 years unmanaged, >50 years unmanaged) influence the radial growth of Fagus sylvatica during fluctuating climatic conditions. On average, trees in managed stands had higher radial growth rate than trees in unmanaged stands during the last two decades a 50%. However, the beech trees in the unmanaged stands were less sensitive to drought than those in the managed stands. This effect was most pronounced in the forest with longest management abandonment (>50 years), indicating that the drought sensitivity of mature beech trees is in these forests the lower, the longer the period since forest management cessation is. Management-mediated modifications in crown size and thus water demand are one likely cause of the observed higher climate sensitivity of beech in the managed stands. Our results indicate a possible trade-off between radial growth rate and drought tolerance of beech. This suggests that reducing stem density for maximizing the radial growth of target trees, as is common practice in managed forests, can increase the trees' drought sensitivity. In the prospect of climate change, more information on the impact of forest management practices on the climate-growth relationships of trees is urgently needed., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Salt marsh plants are affected by regular tidal inundation exposing them to saline water as a potential water source. This study aimed at quantifying the water uptake of plants depending on their distance from the sea and exploring plant responses to changing inundation regimes. We used stable isotope ratios (δ 18 O) to determine the proportions of seawater and precipitation water used by three salt marsh species (Spartina anglica, Atriplex portulacoides and Elytrigia atherica) from a German North Sea coast salt marsh. Additionally, A. portulacoides was transplanted to experimental islands at three elevation levels to investigate its plasticity in water use in the course of future sea level rise. We found a marked gradient in plant seawater use from the lowermost pioneer zone (79-98% seawater uptake by S. anglica) to the lower marsh (61-95% by A. portulacoides) and the upper marsh (25-39% by E. atherica). Seasonal differences in water use were not pronounced, likely due to the absence of longer dry periods during summer in these temperate salt marshes. Contradicting our expectation, roots in deeper soil showed higher water uptake rates per fine root mass than topsoil roots suggesting effective root adaptation to the anoxic subsoil. Transplanted A. portulacoides plants significantly increased the uptake of seawater with increasing inundation indicating flexibility in the use of water sources by this species which may facilitate acclimation to rising sea levels. We conclude that the zonation of salt marsh vegetation reflects the availability of water sources along the inundation gradient.

Climate change can impact forest ecosystem processes via individual tree and community responses. While the importance of land-use legacies in modulating these processes have been increasingly recognised, evidence of former land-use mediated climate-growth relationships remain rare. We analysed how differences in former land-use (i.e. forest continuity) affect the growth response of European beech to climate extremes. Here, using dendrochronological and fine root data, we show that ancient forests (forests with a long forest continuity) and recent forests (forests afforested on former farmland) clearly differ with regard to climate-growth relationships. We found that sensitivity to climatic extremes was lower for trees growing in ancient forests, as reflected by significantly lower growth reductions during adverse climatic conditions. Fine root morphology also differed significantly between the former land-use types: on average, trees with high specific root length (SRL) and specific root area (SRA) and low root tissue density (RTD) were associated with recent forests, whereas the opposite traits were characteristic of ancient forests. Moreover, we found that trees of ancient forests hold a larger fine root system than trees of recent forests. Our results demonstrate that land-use legacy-mediated modifications in the size and morphology of the fine root system act as a mechanism in regulating drought resistance of beech, emphasising the need to consider the 'ecological memory' of forests when assessing or predicting the sensitivity of forest ecosystems to global environmental change.