By selectively exchanging the bound triflate from [OTf⊂NiII4L6](OTf)7 (1), we have been able to prepare a series of host–guest complexes that feature an encapsulated paramagnetic tetrahalometallate ion inside this paramagnetic host giving [MIIX4⊂NiII4L6](OTf)6, where MIIX42− = MnCl42− (2), CoCl42− (5), CoBr42− (6), NiCl42− (7), and CuBr42− (8) or [MIIIX4⊂NiII4L6](OTf)7, where MIIIX4− = FeCl4− (3) and FeBr4− (4). Triflate-to-tetrahalometallate exchange occurs in solution and can also be accomplished through single-crystal-to-single-crystal transformations. Host–guest complexes 1–8 all crystallise as homochiral racemates in monoclinic space groups, wherein the four {NiN6} vertexes within a single Ni4L6 unit possess the same Δ or Λ stereochemistry. Magnetic susceptibility and magnetisation data show that the magnetic exchange between metal ions in the host [NiII4] complex, and between the host and the MX4n− guest, are of comparable magnitude and antiferromagnetic in nature. Theoretically derived values for the magnetic exchange are in close agreement with experiment, revealing that large spin densities on the electronegative X-atoms of particular MX4n− guest molecules lead to stronger host–guest magnetic exchange interactions.Reaction of Ni(OTf) II4L6]8+ cage can reversibly bind paramagnetic MX41/2− guests, inducing magnetic exchange interactions between host and guest.The tetrahedral [Ni CoII) are employed, characterising magnetic properties has not been a key focus.7,8 However, the exploitation of (reversible) guest encapsulation to induce magnetic exchange interactions with the host could be used in a variety of potential applications including magnetic sensing and switching,9 the construction of single-molecule magnets,10 the encapsulation and stabilisation of highly anisotropic single ion magnets with specific geometries,11,12 dilution of magnetic molecules in the solid-state,13 and the organisation of electron spin based qubits within ordered structural frameworks and/or on surfaces.14,15 The latter has proven to be extremely difficult since the magnetic properties of molecules are often changed upon deposition.15The inherent ability of metallosupramolecular cages to encapsulate different chemical species within their cavity can be exploited for a myriad of applications, including the stabilisation of reactive species,i.e. there is no magnetic interaction between host and guest and/or there is no geometrical change in either component. (2) There is no magnetic interaction between host and guest, but binding induces structural changes, altering the geometries of the metal ions in the cage and/or the encapsulated guest, modifying magnetic anisotropy. (3) There is a magnetic interaction between host and guest, which may or may not also change the magnetic exchange between metals ions in the host. (4) There is a combination of points (2) and (3).Introducing a magnetic guest into the cavity of a magnetic host could result in a number of potential outcomes. (1) Guest encapsulation has no effect, II, has shown that even simple solvent molecules can modify the magnetic properties of the metal ions via geometry changes induced by intermolecular interactions. The resulting changes in metal anisotropies can lead to significant changes in magnetisation relaxation dynamics.16 In spin crossover (SCO) MOFs the high spin – low spin transition temperature is well known to be highly guest-dependent, proffering potential application in molecular recognition.17,18 Studies of coordination cages and capsules incorporating a paramagnetic component are limited to the examination of magnetic exchange interactions between metal ions in the cage,19 SCO (of the cage and guest),20–23 and the interaction of organic-radicals in the cavity (with themselves or the cage)24–26 or in the host framework.27,28 The ability to understand, and ultimately control, host–guest magnetic exchange interactions and single ion magnetoanisotropies in such molecular species would represent an important step toward making coordination cages with tuneable, and potentially useful, magnetic properties. Herein, we discuss the construction of a tetrahedral cage [NiII4L6]8+ (L = quaterpyridine) that can (reversibly) bind a range of tetrahedral, paramagnetic MX41/2− guests, inducing magnetic exchange interactions between host and guest.In the chemistry of porous coordination polymers, or metal–organic frameworks (MOFs), the ingress of (non-magnetic) guest molecules into the pores of 3D frameworks built from paramagnetic metal ions, such as CoII4L6 cage was synthesised by combining Ni(OTf)2  with quaterpyridine  in acetonitrile, followed by heating for 24 h 7 (see below). To generate the [MX4⊂NiII4L6]n+ host–guest complexes, the solution obtained after 24 h of heating Ni(OTf)2 and L was treated directly with one equivalent of tetraethylammonium tetrahalometallate salt, giving [MIIX4⊂Ni4L6](OTf)6 where MIIX42− = MnCl42− (2), CoCl42− (5), CoBr42− (6), NiCl42− (7) and CuBr42− (8); [MIIIX4⊂1](OTf)7 where MIIIX4− = FeCl4− (3) and FeBr4− (4). The displacement of the encapsulated triflate is confirmed both by X-ray crystallography (see below) and also by ESI-MS. In this case, ESI-MS . However, as singly charged FeX4−  also displaces the bound triflate, this selectivity is not purely a Coulombic effect, and is likely caused by the shape complementarity of the tetrahedral tetrahalometallate guest for the cage's pseudo-tetrahedral cavity.The tetrahedral Ni2–8 can also be reversibly formed through single crystal to single crystal transformations. For example, when orange crystals of 1 are soaked in an EtOH solution of (Et4N)2NiCl4 for 2 hours, green crystals of 7 are formed. The process is reversed by soaking crystals of 7 in an EtOH solution of nBu4NOTf 7 (1) and [MX4⊂NiII4L6](OTf)6/7 (2–8) were obtained from vapour diffusion of THF and/or Et2O into the MeCN mother liquor. Synchrotron radiation was required to obtain single crystal data for complexes 2 (MX4 = MnCl42−), 7 (MX4 = NiCl42−) and 8 (MX4 = CuCl42−).29 All eight complexes crystallise in monoclinic cells, with 1–4, 7 and 8 being in the space group C2/c, and 5 and 6 in the P21/n and P21/c space groups, respectively –2.17(3) Å and cis/trans angles in the range 76.4(12)–102.28(13)°/170.3(11)–176.6(6)°, respectively. Each tetrahedron has T-symmetry, possessing four metallic vertices with the same stereochemistry (Δ or Λ). All compounds crystallise as racemic mixtures of the homochiral cage .The structures of compounds ix edges . The app1 has a positionally disordered triflate anion occupying the cavity (ortho-pyridyl H atoms of L (O/F⋯H−Ar ≈ 2.52 Å). The remaining seven triflate anions surround the exterior of the tetrahedron, maintaining charge balance. These, and the solvent of crystallisation, are involved in a number of intermolecular interactions that connect neighbouring cages.Consistent with the ESI-MS observations, e cavity . There a7, [NiCl4⊂NiII4L6](OTf)6, is shown in 2–6 and 8, along with pertinent bond lengths and angles, see ESI4, which shows a 1 : 1 partial occupancy of FeBr4− and OTf−, the tetrahalometallate guests are positionally ordered with full occupancy, showing regular tetrahedral geometry. In each case the guest anion is positioned such that the MX4n− tetrahedron is inverted with respect to the cage's [Ni4L6]8+ tetrahedron, i.e. the halide atoms point towards of the portals of the tetrahedron. The host–guest interactions are similar to 1, with the closest contacts between the tetrahalometallate halide atoms and the ortho-pyridyl positions of the cage's ligand. For example, in 7, the distances between the host and guest are: Cl⋯H−Ar ≈ 2.86 Å  with g = 2 for all metal ions, except for NiII where gNi = 2.13 (vide infra). With the exception of 7, on lowering the temperature, the χMT products of 1–8 are essentially constant down to the temperature range 50–20 K, whereupon a further decrease of temperature results in the gradual drop of the χMT products of all complexes, to reach their respective minimum values at 2 K. This behaviour is indicative of weak antiferromagnetic interactions operating in 1–8. The faster drop of the χMT product of 7 from ∼100 K indicates that the tetrahedral NiII guest displays significant magnetic anisotropy, of the same order of magnitude as the temperature.The direct-current (d.c.) molar magnetic susceptibility, 1–8, we performed variable-temperature-variable-field (VTVB) dc magnetisation measurements on polycrystalline samples in the temperature range 2–7 K and in applied magnetic fields up to 7 T. The results of these VTVB measurements are given in the insets of μBB/kT with μB and k the Bohr magneton and Boltzman constant, respectively,  with respect to the experimental conditions . For the quantitative interpretation of the magnetic properties of 1–8, we used spin-Hamiltonian Ĥhost is the spin-Hamiltonian relative to 1, Ĥguest is the Hamiltonian relative to the guests in 2–8 and their interaction with the host 1, i, j are indices that run over the constitutive centres, gNi the g-value of NiII, Ŝi the spin operator of the ith paramagnetic centre, DNi the single-ion axial anisotropy parameter of NiII, SNi = 1 the total spin of NiII, Jij the pairwise isotropic magnetic exchange interaction parameter between centres i and j, with the equivalent quantities for the guests.To better define the low temperature magnetic properties of χMT product and the VTVB data for 1–8 were simultaneously fitted to spin-Hamiltonian 31 For 1, fitting of the χMT product and the VTVB data resulted in the best-fit parameters: gNi = 2.13, |DNi| = 1.575 cm−1 and JNi–Ni = −0.078 cm−1 ; for 4: JNi–Fe = −0.084 cm−1 (DFe was neglected for FeIII); for 5: |DCo| = 2.37 cm−1 and JNi–Co = −0.005 cm−1; for 6: |DCo| = 6.30 cm−1 and JNi–Co = −0.001 cm−1; for 7: |D′Ni| = 85.5 cm−1 and J′Ni–Ni = −0.476 cm−1; and for 8: JNi–Cu = −0.062 cm−1 (DCu was neglected for CuII). The values are tabulated in D′Ni value for the guest in 7 is of the magnitude expected for a tetrahedral NiII ion, and in agreement with the faster drop of the χMT product with decreasing temperature, absent for all other compounds. For these other compounds, the guest magnetic anisotropy is approximately two orders of magnitude smaller, or entirely negligible.The 078 cm−1 . The relab initio methods. DFT calculations have been employed to estimate the isotropic exchange coupling constants (J) using the Hamiltonian . On-site spin–orbit coupling/zero-field splitting (zfs) calculations require accurate estimation of excited state energies, and a single-determinant description of the wavefunction is not sufficient for systems having orbital degeneracies or those possessing low-lying excited states. For this reason, ab initio CASSCF/NEVPT2 calculations have been performed .The magnetic properties of all eight complexes have been investigated using both DFT and JNi–Ni interaction is estimated to be −0.062 cm−1, very close to the experimental value of −0.078 cm−1 , but significant twisting between the bipyridine units  in addition to the JNi–Ni exchange. The spin state energies of the host–guest molecules have been computed considering a pentametallic MnNi4 unit. The JNi–Ni and the JMn–Ni exchange interactions are estimated to be −0.09 cm−1 and −0.01 cm−1, respectively. Here the spin ground state is S = 5/2  in 3 compared to the Mn–Cl distance (2.38 Å) in 2. The CASSCF/NEVPT2 computed D values for the guest ions in complexes 2–4 are small. Indeed, they are smaller than the energy separation between the spin state energies arising from the exchange interaction.Similar analyses were performed on the remaining host–guest complexes. For complex ectively . Note th4 with [FeBr4]− as the guest. In this case the host–guest antiferromagnetic exchange is larger (−0.085 cm−1) than for 2 and 3. The spin density on the FeIII ion is significantly reduced due to strong spin delocalisation on to the electronegative Cl− and Br− ions in 3 and 4  and −3.96 cm−1 (6), three orders of magnitude higher than the energy separation between the exchange-coupled spin states, i.e., |D| > J. The MS level separations, i.e., the gap between the ±3/2 and ±1/2 microstates for CoII are much larger (∼2|D|) and close to 13 cm−1 and 8 cm−1, respectively. Deviation from ideal Td symmetry is well-known to result in a significant D value for tetrahedral CoII ions.35–38A similar situation is observed for complex  Table S6. Interes7, the orbital degeneracy of the tetrahedral NiII ion precluded convergence of the DFT calculations and thus no host–guest JNi–Ni exchange coupling could be estimated. NEVPT2 calculations yield a D value of +214 cm−1 for [NiCl4]2−, a value much larger than that estimated from experimental susceptibility and magnetisation data.39 The origin of this very large anisotropy can be explained from the NEVPT2-LFT orbital splitting pattern of the pseudo-Td [NiCl4]2− complex shown in D value, and arise primarily from the dxy → dyz/dxz and dx2−y2 → dyz/dxz electronic excitations  centres are very small and comparable to the J values (i.e. D ≈ J). For these five species, spin-Hamiltonian For complexes 8, the g-factors for the CuII ion from the NEVPT2 level of theory, gx = 1.633, gy = 2.126 and gz = 4.078 are overestimated. This is a well-known problem in the literature for CuII. Higher electron correlation, higher reference space and/or ligand orbital inclusion, should be taken into consideration.40 We have therefore performed multi-reference CI (MRCI) calculations in combination with the CASSCF wavefunction to obtain the final g-factors, gx = 1.945, gy = 2.470 and gz = 2.879 (giso = 2.43), which remain anisotropic due to mixing with the bromide orbitals. Simulation of the susceptibility and magnetisation data shows excellent agreement with the experimental data . In these two cases, we have used spin-Hamiltonian D values for the CoII ion, ground state effective g-factors for individual Kramers pairs are used as S̃= 1/2 pseudo-spins 7 (1) can be synthesised from the one pot reaction of Ni(OTf)2 and quaterpyridine (L) in acetonitrile. The analogous host–guest complexes, [MIIX4⊂Ni4L6](OTf)6 = MnCl42− (2), CoCl42− (5), CoBr42− (6), NiCl42− (7) and CuBr42− (8), [MIIIX4⊂Ni4L6](OTf)7 = FeCl4− (3) and FeBr4− (4) are formed from 1 by the selective exchange of the encapsulated triflate anion. The complexes can also be formed and interconverted through single-crystal-to-single-crystal transformations.The tetrahedral cage [OTf⊂Ni4n− guest molecules leads to stronger host–guest magnetic exchange interactions. For the tetrahedral CoII guests, the anisotropy is small but still much larger than the magnitude of exchange coupling between host–host and host–guest. The orbital degeneracy of the tetrahedral NiII ion and the very large zfs that results makes accurate estimation of JNi–Ni and DNi(tet) in (7) rather difficult, as reflected in the large differences in the results obtained between experiment and theory.Magnetic susceptibility and magnetisation data show that the magnetic exchange interactions between metal ions in the host complex, and between host and guest, are of comparable magnitude and antiferromagnetic in nature. Theoretically derived values for the exchange are in close agreement with experiment and reveal that large spin densities on the electronegative X-atoms of certain MX43,44What is clear, however, is that the encapsulation of paramagnetic guests inside dia/paramagnetic cages can be very useful in an array of potential applications. These include sensing and switching, the encapsulation and stabilisation of highly anisotropic (and/or air- and moisture-sensitive) magnetic molecules and the organisation (and/or dilution) of magnetic molecules within ordered, solution-stable structural matrices. To date, surface deposition of magnetic molecules has proved problematic, since in the vast majority of cases structural/magnetic integrity is compromised hindering application. Encapsulation of metal complexes such as spin crossover species, single-ion magnets (SIMs) or electron spin based qubits within a dia/paramagnetic cage whose exohedral organic skeleton is easily derivatised may prove to be an interesting option. The cage acting both as a surface anchor and a protective coating for the magnetic molecule.4n− guests were observed here, this work also suggests that the deliberate distortion/construction of magnetic molecules through encapsulation within the confines of a sterically restricted cavity of a coordination cage may offer an alternative route to producing highly unusual/anisotropic SIMs with specific geometries, tailored ligand fields, and targeted symmetries. In turn, such species may display a breadth of fascinating structures and magnetisation relaxation dynamics that may not exist outwith the cage.Although only relatively small changes to the geometries of the MXAJS and JV performed the experimental work. AS and GP performed the theoretical studes. AJS, JV, LS, ERM, MM and SP measured the magnetic data. SP and AJS fitted the magentic data. WTK and SJC collected and solved the single crystal X-ray diffraction data. PJL and EKB conceived the idea. AJS, GR, SP, PJL and EKB wrote, reviewed and edited the manuscript.There are no conflicts to declare.SC-012-D1SC00647A-s001SC-012-D1SC00647A-s002


Understanding the structure and composition of landscapes can empower agencies to effectively manage public lands for multiple uses while sustaining land health. Many landscape metrics exist, but they are not often used in public land decision-making. Our objectives were to (1) develop and (2) apply a process for identifying a core set of indicators that public land managers can use to understand landscape-level resource patterns on and around public lands. We first developed a process for identifying indicators that are grounded in policy, feasible to quantify using existing data and resources, and useful for managers. We surveyed landscape monitoring efforts by other agencies, gathered science and agency input on monitoring goals, and quantified the prevalence of potential indicators in agency land health standards to identify five landscape indicators: amount, distribution, patch size, structural connectivity, and diversity of vegetation types. We then conducted pilot applications in four bureau of land management (BLM) field offices in Arizona, California, and Colorado to refine procedures for quantifying the indicators and assess the utility of the indicators for managers. Results highlighted the dominance of upland and the limited extent of riparian/wetland vegetation communities, moderate connectivity of priority vegetation patches, and lower diversity of native vegetation types on BLM compared to non-BLM lands. Agency staff can use the indicators to inform the development of quantitative resource management objectives in land use plans, evaluate progress in meeting those objectives, quantify potential impacts of proposed actions, and as a foundation for an all-lands approach to landscape-level management across public lands. Five landscape indicators are relevant to existing policy for multiple-use public lands.We quantified the indicators for priority vegetation types in three western states.Landscape indicators can inform objectives in land use plans for public lands.Indicator products must fit agency workflows and reporting requirements to be used.Limitations of current landcover data impede landscape monitoring on public lands. There is a growing understanding that successful, long-term resource management and conservation often require prioritizing, coordinating, and implementing actions at landscape scales , public lands are extensive and have complex and intertwined land ownership patterns . Many public lands are managed for multiple uses  and are under pressure to provide diverse products and values to society, including oil and natural gas and diverse recreational opportunities , while others affect resource patterns and conditions across large areas and over longer time periods . Disturbances large and small, and their associated direct and indirect effects on people, wildlife, and resources at multiple spatial and temporal scales, all contribute to cumulative effects of past, present, and reasonably foreseeable future actions, land uses, and associated human impacts that can significantly alter landscapes  develop and (2) apply a process for identifying a core set of landscape indicators that public land managers could use to assess, and ultimately monitor, resource patterns within and across landscapes in the western US. We define core indicators as indicators that quantify measurable ecosystem components that are applicable across ecosystems, management objectives, and agencies BLM . We defiBelow we first present information on the process we developed for identifying policy-relevant landscape indicators, and the application of that process to the BLM. We then describe methods, results, and agency feedback on the four pilot applications. Finally, we discuss our process, datasets, and methods along with the importance of science-management partnerships in efforts such as this one that attempts to bridge the gap between science and its use in public lands decision-making. We close by identifying specific ways in which these indicators can be used to inform future planning and management decisions on multiple-use public lands.We began our effort to identify core landscape indicators by holding a scoping workshop with a group of 42 science and technical advisors to generate an initial outline of a process to identify the indicators, and then followed up with participants and other agency staff to solidify the key steps, information needs, and desired products for public land managers. This process was modeled after those used previously by the BLM to identify core indicators for field-based monitoring of terrestrial and lotic systems . Documenting clear references to landscape-level resource management goals and reporting needs in existing agency laws and policies is critical for identifying indicators that will be relevant to agency work. Indicators that can inform or facilitate required reporting are most likely to be viewed as useful by agency staff.Step 3 is to quantify the proposed indicators in pilot applications and gather input from agency staff to help ensure that the indicators are feasible to quantify, clearly presented to staff in easy to use formats and products, and relevant to agency planning and management across different geographies, ecosystems, and management contexts. Steps 2 and 3 are both likely to be iterative.Two foundations for the effort were articulated by our project advisors. First, because many changes in public lands policy were occurring at the time, advisors emphasized that the indicators needed to be grounded in foundational laws that are unlikely to be altered as elected officials and their administrations change over time. Accordingly, we identified the Federal Land Policy and Management Act of 1976, the Fundamentals of Rangeland Health, and the National Environmental Policy Act [USC §4321] as foundational to BLM resource management.Second, advisors recommended eight guiding principles for the effort which focused on relevance to BLM reporting and simplicity in terms of concept and interpretability Table . The advWe also explored if and how other organizations are assessing and monitoring natural resources at landscape scales. A few clear patterns and lessons emerged. First, multiple organizations have taken steps toward landscape-level assessment and monitoring of vegetation patterns, including the Environmental Protection Agency  using datasets likely to provide informative results over timeframes coinciding with required land use plan reporting (5–10 years).We identified BLM land health standards and indicators as the key mechanisms most directly connecting BLM policy to landscape concepts. BLM land health standards are derived from the Fundamentals of Rangeland Health (43 CFR §4180): (1) watersheds in properly functioning condition, 2) ecological processes that support healthy biotic populations and communities; (3) water quality that complies with state standards and achieves BLM objectives; and (4) habitats that support special status species. From these fundamentals, the BLM worked with 19 Resource Advisory Committees to develop land health standards and indicators for different geographic areas , land health standards and indicators for the area, and input from field and state office staff  to identify priority vegetation types and relevant vegetation management objectives for each pilot application. BLM resource management plans are comprehensive documents that identify, among many other things, desired outcomes  for vegetative resources, including desired mixes of vegetative types and structural stages; landscape and riparian functions; habitat for native plants, fish, and wildlife; and forage for livestock. Desired outcomes may be established at multiple scales. Plans may also identify areas of ecological importance, designate priority plant species and habitats, and identify actions and area-wide use restrictions needed to achieve desired vegetative conditions BLM .We evaluated three datasets as sources for current vegetation: LANDFIRE Existing Vegetation Type  within 500 m of a focal pixel. We first computed and mapped current vegetation type diversity in line with our monitoring objective to assess current status. Based on requests from BLM staff, we also computed and mapped the difference between current and estimated historic vegetation type diversity, using the difference between the count of class names for natural vegetation types between EVT and BpS for each neighborhood.We first present results for the Yuma Field Office and then summarize results across the four study areas. Results for the Hassayampa, Eagle Lake, and White River field offices are provided in Appendices A, B, and C, respectively. For all study areas, we illustrate patch metrics for one common upland priority vegetation community and for riparian/wetland, a scarce lowland vegetation community that is always a priority for BLM managers.The Yuma Field Office resource management plan BLM  identifiCoccyzus americanus] in conjunction with removal of competing for exotic species ”. Our analysis can be used to partially evaluate this objective by considering the frequency distribution of patch sizes of all riparian/wetland vegetation.Across our study areas, we identified between six and 12 priority vegetation communities based on the most recent resource management plans and amendments and contemporary issues noted by field staff. We were able to map and quantify the majority of those communities, but some were not well characterized by LANDFIRE EVT classes, and the area of a few communities was too small for meaningful analysis of landscape patterns. Notably, only one plan had a  quantitative vegetation management objective related to landscape patterns. The Yuma plan included the objective: “VM-013—Manage for large, contiguous blocks of native riparian habitat (>30 acres) for yellow-billed cuckoo [2 or smaller in size, however, the total area of creosote white bursage is relatively evenly distributed across three patch-size classes  in both the field office and the state, with no clear difference between BLM and non-BLM lands . However, there are always far more small patches (<10 km2) in terms of absolute numbers  in size, but only 2% of individual riparian/wetland patches are this large  in the Eagle Lake Field Office and northwest Great Basin on both BLM lands  and non-BLM lands . However, there are always far more very small patches (≤0.02 km2) of riparian/wetland vegetation in terms of absolute numbers , while the BLM field offices in California and Colorado manage lands across a range of diversities  in the northwestern Great Basin). Staff were concerned about the accuracy of vegetation data in their field office and identified this as a major challenge to broader implementation and regular use of landscape metrics in decision-making.As part of the pilot applications , we worked closely with BLM state and field office staff and gathered informal feedback from them on both the indicators and the quantitative results of our analyses in their region over the course of multiple meetings and work sessions. BLM staff across levels and positions appreciated the baseline information provided by the indicators for their priority vegetation communities and considered it to be directly relevant to (and in a format that can now be easily integrated into) their land use plans. The indicators afford them a better understanding of the current status of their resources, which they can use to identify vegetation treatment and restoration needs. For example, the limited amount of riparian/wetland vegetation managed by the BLM was expected based on patterns of settlement in the western US in which lands along streams were primarily secured by settlers  and on anthropogenic and ecological processes that affect resources . Many vegetation types that are priorities in land use plans are priorities because they are a rare or minor component of the landscape. One goal of the comprehensive remap of LANDFIRE EVT, which resulted in LANDIRE 2.0, was to increase classification accuracy for some of these vegetation types, including riparian , November 2018). The large units used for assessing the accuracy of LANDFIRE EVT, while ecologically based and appropriate for a nationwide dataset and the limited number of available testing data for each vegetation type, are part of this challenge. Agency land managers would be more convinced of the quality of the data if accuracy was assessed at the smaller extents  at which they are likely to use the indicators. Another federal land management agency in the US, the National Park Service, decided to use a different landcover dataset . We suggest that the methods and specific metrics used here to be viewed as a starting point. We hope that as managers become familiar and comfortable with these landscape concepts, they will want additional and more nuanced information about the landscape-level patterns of the resources they manage. Connectivity, in particular, is an area of focus currently among both federal and state resource managers (US Department of the Interior Our goal was to identify indicators relevant to public lands policy and demonstrate their relevance to and feasibility for use in public land management. We believe involvement and input from a broad group of stakeholders and agency staff throughout this project were a core strength of the process we developed to identify policy-relevant landscape indicators. We have highlighted above a number of challenges to their regular use, including the availability of landcover datasets with an adequate thematic resolution that is produced at regular intervals using consistent methods. Limited staff time and agency expertise in landscape ecology can also hinder the use of landscape information in public lands decisions, as can the short timeframes in which many decisions must be made (e.g., Trammell et al. We suggest that these core landscape indicators can help to improve understanding, management of, and reporting on the health of multiple-use public lands in five main ways. First, the indicators provide a viable method for BLM to quantify existing landscape-level land health standards and indicators. BLM is required to manage public lands to meet these land health standards and recommends that land health be assessed at broad scales (e.g., watersheds, BLM Supplementary Information


To avoid the influence of coarse aggregate, a qualitative estimate of the amount of coarse aggregate analyzed has been made, although the results show that there is no significant influence. Monte Carlo simulations were carried out in order to establish the necessary number of random analyses of the mean value to be within an acceptable range of error. In the case of quantifying sulphates, it is necessary to carry out six random analyses on the surface, and eight measurements in the case of quantifying chlorides; in this way, it is ensured that errors are below 10% in 95% of the cases. The results of the study highlight that a portable XRF device can be used in situ to obtain concentrations of chlorides and sulphates of a concrete surface with good accuracy. There is no need to take samples and bring them to a laboratory, allowing lower overall costs in inspection and reparation works.A correct assessment of the pathologies that can affect a reinforced concrete structure is required in order to define the repair procedure. This work addresses the challenge of quantifying chlorides and sulphates directly on the surface of concrete. The quantification was carried out by means of X-ray fluorescence analysis on the surface of concrete specimens at different points with portable equipment. Concrete prisms were made with different amounts of NaCl and Na The sulphates present in mortars and concretes may come from the gypsum or anhydrite added to the clinker in the cement manufacturing process as a setting retarder, or they may have been incorporated once cement has hydrated and hardened. An excess of sulphates can be harmful, since if they react with hydrated calcium aluminates, secondary ettringite is formed, while they react with CSH gel, thaumasite can be formed ,2,3. TheHowever, the sulphates present in mortar or concrete may not be part of these degradation products; rather, they may be adsorbed on other solid compounds or be part of the pore solution. These sulphates will not be harmful if they are not mobilized and react with the compounds mentioned above. There are also cements that, due to their lower calcium aluminate content and/or the presence of substituent additions, are more resistant to sulphates .For all these reasons, there are no sulphate concentration thresholds from which it can be deduced that if these values are exceeded, there is a sulphate attack. Quantifying these ions should be taken merely as an indication and should be complemented by a broader study. Other types of tests on the mortar or concrete are necessary to be able to conclude the existence of an attack, usually scanning electron microscopy (SEM) and X-ray diffraction (XRD) ,7 togethOn the other hand, chloride ions are one of the main causes of deterioration of reinforced concrete structures, causing corrosion of the reinforcement. Chloride causes the breakage of the passive layer that protects the steels, exposing them to further oxidation ,11,12. CAlthough there is no agreement on a limit of chloride concentration in concrete that causes corrosion of reinforcement ,22, inteThere are different methods of sulphate quantification. Standard methods involve dissolving the solid sample and precipitating the sulphates using a barium salt and then gravimetrically quantifying the initial sulphates in the sample. This methodology requires an adequate chemical analysis laboratory and the analysis is tedious and time consuming. Its reliability or reproducibility is acceptable .The reference method for the quantification of chlorides is based on the Volhard method, by which the chlorides present in a solution are precipitated by the addition of a silver salt to form an insoluble silver chloride . The det3 in a solid sample, but it is also capable of quantifying the specific signal of the chlorides present in a cementitious matrix. It should be noted that, as stated in the standards, all quantification methods other than those mentioned above as standardized require prior calibration [X-ray fluorescence (XRF) is a spectroscopic technique for elemental quantification. Fluorescence is a physical process that occurs in response to radiation by high-energy beams at a solid sample and occurs as a consequence of electronic relaxation when such radiation disappears. The response is element specific and proportional to its concentration in the sample. This technique can be used to calculate the sulphur concentration expressed as SOibration .Handheld X-ray fluorescence equipment (pXRF) allow this technique to be used for elemental analysis in works, buildings, facilities, and infrastructures far from the laboratory, since the analyzer is a small and lightweight device . On the Handheld X-ray equipment (pXRF) generally comes with an internal calibration that allows the quantification of elements in a semi-quantitative way, but as in any instrumental technique, it is advisable to calibrate the equipment to determine the desired elements in the matrix in which they are found.During the inspection of a work, installation, or infrastructure, it is very useful to know at the moment the cause of any apparent degradation of concrete. Normally, when it is necessary to know the elemental composition of the concrete, samples have to be taken and sent to a specialized laboratory for analysis. When the objective is even more specific, such as precisely delimiting the most affected areas in order to define and repair with greater objectivity, the increase in the time required for the entire process significantly increases the costs of these repair stages.The purpose of this study is to use pXRF equipment to directly quantify the concentration of chlorides and sulphates on the surface of concrete. Given the large number of variables that can influence this concentration, a previous experimental campaign has been carried out in which several concrete specimens with known concentrations of these ions have been analyzed in the laboratory. A large number of punctual analyses have been carried out in order to know whether their average value is close to the real value. Subsequently, by means of a Monte Carlo simulation, the number of punctual analyses necessary for their mean value to be as close as possible to the real value within an acceptable error has been obtained.Monte Carlo simulation is a statistical analytical method used to mathematically model real systems and estimate the probability of obtaining the expected outcome ,28. SincThe pseudo-population or model that represents the true population of interest is determined.Samples are taken from this pseudo-population.A value of the statistic parameter of interest is calculated and stored.N trials.Repeat steps 2 and 3 for N found in Step 4 are used to study the distribution of the statistic parameter.Finally, the values of The computer software for managing the pXRF shows the spot or analysis region where the analysis is carried out see . The phoThe graphical representation of the concentrations obtained as a function of the factor allows knowledge of the influence of the coarse aggregate in the quantification by this technique, since the main sources of sulphates and chlorides are found in the cementitious matrix.-Identification of the optimal distribution of sulphate and chloride quantification results. Once the distribution is defined, the descriptive statistic parameters for each one of them are defined.-In the simulation itself, random values are taken for each of the distributions defined by the previous statistic parameters. The number of random values is increased so that when computing the averages, it is possible to identify which average is close to the real value with an error of 10%.Once the influence of the aggregate was verified, a simulation was carried out using the Monte Carlo method to determine the number of random point analyses that achieve an average value as close as possible to the real value. This simulation followed the following process:2SO4 or NaCl (dissolved in the mixing water) was introduced into each of the specimens.Concrete specimens of dimensions 15 cm × 15 cm × 15 cm were made. A known percentage of Na3 and a cement quantity of 350 Kg/m3 in all cases , the soils mode was selected. The quantification results are expressed in parts per million (ppm). Chlorides are quantified directly as chloride ions, but sulphate quantification is expressed as total sulphur (S) and a conversion to SOA workstation from the same manufacturer was also available to connect the pXRF equipment and analyze the samples without the analyst being subjected to radiation or having to hold the equipment manually during the analysis. The workstation was connected to a computer to monitor the tests from the computer. Thanks to the fact that the analysis interface of the equipment’s software allows us to observe the area being analyzed, we can easily detect whether we are in an area with more or less aggregate. Each analysis was categorized from 0 to 4 according to the amount of aggregate, with 0 being all paste (and fine aggregate) and 4 being an analysis performed on 100% coarse aggregate.Although the pXRF equipment has an internal calibration, prior to the concrete surface analyses, samples with different amounts of sulphates and chlorides were prepared to calibrate perfectly the response of the instrument to samples whose matrix was similar to the object of analysis. The results of this calibration with powder samples of test concretes  indicatex) and standard deviation (sdx). Distributions will be obtained for each analyzed element and its concentration in the concrete.Fit the experimental data to an appropriate distribution and obtain the characteristic values, such as averages  and the distribution to which the experimental data fit, it is possible to obtain the optimum number of analyses to be performed on a concrete surface to be within an acceptable error with minimum uncertainty. The process follows the scheme below:4 concentration of 0%, As can be seen in the box and whisker plots in The following box-and-whisker plots show the chloride concentration as a function of the aggregate occupancy factor. It was decided not to omit the outliers; of which it should be noted that they are always on the high concentration side. This could be due to a higher point concentration on the specific surface of analysis or to recrystallization by evaporation in concrete pores. The number of observations for the 0% chloride specimen is significantly lower than for the rest of the specimens, since a large number of spot analyses gave quantification results below the detection limit of the technique.In view of the results in In order to perform an optimal simulation of the sample, it is necessary to know the distribution of the sample and to use the representative statistics of this distribution. The histograms of each of the samples are shown graphically below, together with the representation of the normal and lognormal density functions for each of them. These representations show which distribution best fits each of the histograms represented.The statistic parameters that represent a normal distribution are the average and the standard deviation. In the case of the sulphate quantification experiments, these are shown in Histograms of the chloride quantification results of the different test specimens are shown in A random value is taken that meets the conditions for each of the distributions; that is, they are within a normal distribution with the mean and standard deviation of 2 and sd2) are saved.Next, we simulate taking two values with the same conditions as in Step 1 and calculate the mean and standard deviation. The process is replicated one million times and the mean and standard deviation values .The same process is simulated for 3, 4, 5, 6, … up to 10 values. For each one, it is replicated one million times and the mean and standard deviation values are saved SE = sdn + 1.96 * SEIC 95% = meann + 1.645 * SEIC 90% = meann ± 0.1 * meannError ±10% = meanRandom values within the distributions found were then sampled. These random values correspond to the number of point analyses on the concrete surface. For the Monte Carlo simulation, the following process was followed:To check the number of point analyses on a concrete surface to be carried out with the portable X-ray equipment, it is sufficient to check the value of x at the intersection of the 95% IC (or 90% IC) curves with the 10% error on the graphs obtained.The results of the Monte Carlo simulation of sulphate quantification on the surfaces of the different concrete specimens are shown in As can be seen in The results of the simulation for the case of chloride quantification are shown in In view of the simulation results, it can be concluded that with about eight random measurements  on the surface of a concrete, we are quantifying chlorides within a ±10% error with a confidence interval of 95%, or in other words, in only 5% of the cases we cannot be sure of having an error of less than 10%.The study presents the results of an experiment in the quantification of chlorides and sulphates in concrete using handheld X-ray fluorescence equipment (pXRF). Calibration for quantification directly on the surface is shown, without the need for sample extraction or sample preparation.It has been observed that the concentration of these ions does not depend on the amount of coarse aggregate analyzed, since the dispersion around the mean value is much greater than the effect of the aggregate itself. The distribution of the quantification results for sulphate conforms to a normal density function, while chlorides conform to a lognormal one.n values is within the 10% error in 95% and 90% of the occasions, with being n the number of spot analyses to be done with the pXRF device on the surface of the concrete.A simulation was carried out using the Monte Carlo method based on the mean and standard deviation statistics of the sulphate concentrations in each of the specimens and the medianlog and sdlog values in the case of chlorides. The Monte Carlo method allows knowledge of what mean of The results show that by performing nine analyses on the surface of the concrete, the chloride content can be determined with an error of less than 10% at a 95% confidence interval. In relation to sulphates, the results of the study indicate that with a number of six analyses, we can obtain its content with an error of less than 10% at a 95% confidence interval.Although the quantification analyses obtained using the pXRF equipment are not as accurate as traditional X-ray fluorescence, the results are acceptable if we take into account the necessary prior calibration for each piece of equipment and its use in cement-based materials.


According to the social exchange theory, this study analyzed how a bottom-line mentality (BLM) among leaders affects teachers' innovative behavior and how this relationship is mediated by relative deprivation and psychological safety and moderated by person-organization values fit. Using two stages of data collection, 491 responses from teachers were obtained and analyzed. The results revealed that leader BLM significantly negatively affected teachers' innovative behavior, and relative deprivation and psychological safety both partially mediated this influence of leader BLM. Person-organization values fit negatively moderated the positive effect of leader BLM on teachers' relative deprivation and the negative effect of leader BLM on teachers' psychological safety. This study enriches the current literature about BLM and tests the influence of leader BLM on teacher's innovation in the Chinese education and training institutions, and provides insights into favorable educational management practices. Education is a process of teaching others to think with knowledge as a tool, thinking about how to create higher social wealth and realize the embodiment of self-worth. In the global competitive environment, how to practice good education is very important to cultivate innovative talents. Based on the restriction of public financial investment, the main direction of public education is to provide more and better basic education public services, while private education can provide more differentiated education services with high quality to meet people's individualized education needs. Only when public education and private education complement each other and cooperate with each other can we resolve the main contradictions in the field of education and better meet people's growing educational needs. Whether it is education and training institutions, or the operation of any category, market models and laws are changing with the development of the times and the progress of science and technology. Thus, innovation is undoubtedly the most important thing. Also, in the highly competitive education industry, the educational innovation practice and reform of the education and training institutions undoubtedly become important ways to obtain sustained competitive advantages Senior, . With thbottom-line mentality (BLM). According to the research theme, we define leader BLM as the stubborn thinking that the administrators of educational and training institutions only pay attention to organizational interests and personal affairs, while ignoring the professional development of teachers and trainers in the institutions. In corporate, the obsession of management with bottom-line goals has led to various corporate scandals around the world. It's worth noting that in Chinese education industry, similar education scandals also exist and continue to occur. How to maintain the advancement and purity of Chinese education industry is a major issue to be considered by academia and education industry. Therefore, research on the BLM of leaders in Chinese education context can provide theoretical and practical implications to education practice in China. Studies have demonstrated that leader BLM has a certain negative effect on individuals, such as causing unethical  have been a focus in recent studies. Although both qualitative and quantitative studies have revealed the negative effects of BLM on organizations and individuals, few studies have discussed the influence of leader BLM on individual constructive behavior, and most studies have been conducted from a single perspective . Also, BLM is a worldwide phenomenon, but studies on leader BLM in the context of Chinese management are few, which has been highlighted by scholars promoting leadership localization  to 5 (completely agree). Teachers of the English language and management were recruited to conduct back translation; subsequently, the translated scales were modified by two teachers of management and psychology with reference to the original scales before finalizing the Chinese version of the scales  and composite reliability (CR) to assess the reliability of each scale. 2 = 397.045, df = 179, χ2/df = 2.218, comparative fit index = 0.946, incremental fit index = 0.947, Tucker–Lewis index = 0.937, root-mean-square residual = 0.044, root-mean-square error of approximation = 0.050) is considerably higher than that of the other models, indicating high discriminant validity in the five factors  and psychological safety , and significantly positively correlated with teachers' relative deprivation . Teachers' psychological safety and innovative behavior are significantly positively correlated . Teachers' relative deprivation and innovative behavior are significantly negatively correlated . These correlation results provided a basis for the subsequent analyses.This study conducted regression analysis and concurrently tested the multicollinearity of the variables. Variance inflation factor analysis revealed that the variance inflation factor was lower than two for all variables, indicating no multicollinearity in the research model.p < 0.001, M6); thus H1 was supported. Besides, we also found that leader BLM has a significant negative effect on teacher's psychological safety , and has a significant positive effect on teacher's relative deprivation . At last, as we can see in the p < 0.001, M7), and the relative deprivation has a significant negative effect on their innovation . The test of direct effects provide a basis for further analysis.Regression analysis was conducted using SPSS22.0 with control of the demographic variables. p < 0.01, M10); relative deprivation has a significant negative effect on teacher innovative behavior . Accordingly, relative deprivation has a partial mediating effect on the relationship between BLM and teacher innovative behavior, supporting H2a. To test the mediating effect of psychological safety, BLM and psychological safety were used as independent variables and innovative behavior as the dependent variable. The regression analysis revealed a significant negative effect of BLM on teacher innovation behavior, but the regression coefficient is reduced ; psychological safety exerts a significant positive effect on innovative behavior . Accordingly, psychological safety exerts a partial mediating effect on the relationship between BLM and teacher innovation behavior, supporting H2b.A method proposed by Baron and Kenny  was usedp < 0.01, M3), indicating that P-OVF negatively moderates the relationship between BLM and relative deprivation, partially supporting H3a. The BLM, P-OVF, and interaction term were employed as the dependent variable and psychological safety as the independent variable in another regression analysis. The interaction term has a significant positive effect on psychological safety , indicating that P-OVF negatively moderates the negatively relationship between BLM and psychological safety, partially supporting H3b (Subsequently, the moderating effect of P-OVF was tested. Before the test, leader BLM and teachers' P-OVF were both centered, and their interaction term was obtained. The BLM, P-OVF, and interaction term were employed as independent variables and relative deprivation as the dependent variable in regression analysis. Seen as ting H3b . SpecifiFinally, the PROCESS macro for SPSS was used in conjunction with the bootstrap procedure to test the moderated mediation Hayes, . InnovatInnovation is a crucial means with which the education and training industry achieve a competitive advantage and has thus received considerable research attention. To analyze the causal relationship between leader BLM and teacher innovation behavior, this study constructed a theoretical model comprising leader BLM, relative deprivation, psychological safety, and teacher innovative behavior and incorporated P-OVF as a moderating variable. The causal relationships, mediating effects, and moderating effects among these variables were analyzed. The obtained results supported the proposed hypothesis.Leader BLM significantly negatively affects teacher innovation behavior. This result indicates that the prioritization of only education and training industry profit by leaders results in less innovative behaviors of teachers during work, tasks, and interactions with colleagues. Relative deprivation and psychological safety both mediate the relationship between leader BLM and teacher innovation behavior. Thus, leader BLM reduces the amount of teacher innovation behavior through its effect on teachers' psychological perceptions. Finally, P-OVF exhibits a moderating effect. Specifically, favorable P-OVF is associated with a weak positive effect of leader BLM on teachers' relative deprivation and concurrently moderates the mediating effect of relative deprivation. Furthermore, favorable P-OVF results in a weaker negative effect of leader BLM on teachers' psychological safety and moderates the mediating effect of psychological safety.The theoretical significance of this study is mainly reflected in the following aspects: First, we investigates the influence mechanism of leaders' bottom-line mentality on teachers in the Chinese educational context. Leaders' bottom-line mentality is the research frontier in organizational behavior, especially in the context of globalization that pays attention to the quality of economic development. Keeler and Webster  pointed Secondly, following the research on leaders' bottom-line mentality, it is found that although the bottom-line has played a certain role in ensuring economic profits, scholars have found that if they only pay attention to corporate profits and personal interests, there will be a series of adverse consequences (Sims and Brinkman, Third, relative deprivation and psychological safety played partial mediating roles between leaders' bottom-line mentality and teachers' innovative behavior. First of all, the results of this study clarify the specific impact mechanism of leaders' bottom-line mentality on individuals' innovative behavior, and at the same time expand the theoretical perspective of individuals' pre-factors of innovation. Although previous studies have confirmed the influence of leaders' bottom-line mentality on individuals' behavior, they are mainly based on the perspective of leadership identity and moral identity (Greenbaum et al., Finally, the person-organization values fit has a moderating effect. In previous studies, when the bottom-line mentality of leaders influenced the subordinates, most scholars took moral identity (Mesdaghinia et al., The main purpose of this research is, by referencing the research conclusions, to provide recommendations related to routine education management practices for education and training institutions. Based on the present research conclusions, this study proposes corresponding strategies aimed at sustaining teacher innovation and long-term school education development.The present study and relevant studies have demonstrated that leader BLM exerts a negative effect on psychological and behavioral aspects of individuals (Greenbaum et al., We find that relative deprivation and psychological safety partly mediate the relationship between leader BLM and teachers' innovative behavior, which makes us have to put forward some suggestions to effectively avoid the negative impact of leader BLM. First of all, we need to find some effective measures to reduce teachers' sense of relative deprivation and improve their psychological safety, so as to alleviate the transmission of negative effects to innovative behaviors. For example, education and training institutions can increase the welfare of teachers, which has been proved to be an effective way to alleviate unfair perception in some literatures. In addition, these institutions can also practice appropriate human resource management to improve individual psychological safety, such as developmental human resource management, inclusive human resource management and committed human resource management. Secondly, partial mediation shows that we can try other ways to transform the negative effects of leader BLM on individual innovation. For example, leader BLM pays more attention to organizational benefits, so we can take appropriate measures to encourage teachers to form performance-oriented approach to goals, such as advocating collective values, which can promote their innovative behavior under the leadership of BLM.What's more, education and training institutions must acknowledge the importance of innovative teacher behavior in education and training institutions' education development. Education and training institutions can construct an internal learning platform that connects with external platforms to create a learning atmosphere, encourage the formation of learning groups, and thus elicit innovative teacher behavior. This kind of learning method can stimulate teachers' creativity and improve educational practice and teaching quality. Hands-on education activities are essential to fostering innovative teacher behavior; therefore, education and training institutions can hold regular innovation competitions, teaching contest, and educational quality development activities to engage teachers in innovation practices, train their innovative education thinking, and thereby drive their innovative behavior.Despite obtaining valuable research results, this study had some limitations and there was room for improvement. First, this study investigated only how leader BLM affects teacher innovation behavior; the effect of leader BLM on organizations and other teacher behaviors was not addressed. Accordingly, future research may examine the causal relationship of leader BLM with other factors such as innovation atmosphere in the organization, teachers' organizational commitment, and job burnout. Additionally, this study explained the influence of leader BLM on innovative teacher behavior from the perspective of psychological safety and relative deprivation, but other mediators may also exist. Therefore, scholars can explore the effect from different perspectives to uncover potential mediators. Finally, this study analyzed the moderating effect of P-OVF; however, whether the causal effect of leader BLM is affected by other situational factors has not been determined. Accordingly, other boundary conditions, such as leaders' identity, could be incorporated.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by the Academic Committee of Business School of Huaqiao University. The patients/participants provided their written informed consent to participate in this study.This study was a joint work of the five authors. LL, WW, and YW contributed to the ideas of educational research, collection of data, and empirical analysis. LL, JL, WW, and QF contributed to the data analysis, design of research methods, and tables. LL, WW, JL, and YW participated in developing a research design, writing, and interpreting the analysis. All authors participated in reading and approval of the final manuscript and have read and agreed to the published version of the manuscript.The 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.


PaleoClim representing warm and cold cycles; and (2) from present to the year 2100 (CSI-future), using nine general circulation models of climate change of four periods available from WorldClim. We calculated standard deviation of the variables and selected an uncorrelated set for summing, normalizing and obtaining the CSI maps. Our approach is useful for fields such as biogeography, earth sciences, agriculture, or sociology. However, CSI is an index that can be re-calculated according to particular criteria and objectives (e.g. temperature variables); maps are, therefore, customizable to every user.Climate changes are top biodiversity shapers, both during the past and future. Mapping the most climatic stable and unstable zones on Earth could improve our understanding of biodiversity distribution and evolution. Here, we present a set of maps based on a global scale, high resolution (ca. 5 km) new Climate Stability Index (CSI). The CSI considers bioclimatic variables for two different time ranges: (1) from Pliocene (3.3 Ma) to the present (CSI-past map set), using 12 time periods of  Machine-accessible metadata file describing the reported data: 10.6084/m9.figshare.19050569 In contrast, regions with relatively stable climates have often acted either as “museums” (places of persistence)5 or both as “museums” and “cradles”7. In addition, recent global warming is causing even deeper changes in biodiversity in a very short space of time. Increases of temperature in just a few decades might produce large regional species turnover by increasing extinction rates and large migrations8, and also by loss of phylogenetic diversity 9.Long-term climatic variation has had an enormous impact on the evolution of biodiversity, including humans. Climatic instability linked to the Pleistocene glacial/interglacial cycles has caused active diversification10, following the demand of incorporating evolutionary processes into conservation planning11. Mapping climatic stability for the future can be even more important, as climate patterns are predicted to change dramatically during this century, including the occurrence of extreme weather events12. Such mapping may have, thus, implications on economy and human health; e.g., when selecting new arable lands or new human settlements that will be needed as a consequence of the predicted large-scale migrations8.Identifying areas with high climatic stability is of enormous interest. Delimiting past trends of stability and mapping them can help to shed light into evolutionary processes that shaped current biota, including human evolution. They may also be helpful in designing new protected areas (PAs) and prioritizing or redesigning the existing onesStableClim13 and the “map of climate stability”14 (hereinafter “MCS”), which are projections of climate variation, the former from 21,000 BP  to 2100 CE13, and the latter from 21,000 BP to 100 BP14. StableClim offers estimates of climate stability on an ideal time-scale (every 100 years) for surveying the eco-evolutionary impacts of short-term climate shifts13, but it shares with MCS three limitations14: (1) coverage of only ca. 21,000 years, which hinders their utility for explaining the role of climate stability in evolution and speciation that take thousands or millions of years; (2) spatial resolution is insufficient , which does not allow one to correlate climatic stability with population data on both local and regional scales; and (3) they use one or two variables (mean monthly temperatures13 or mean monthly temperatures and precipitations14) that may not be the key ones for a given case of study; for example, distribution patterns of bees in South Africa are highly dependent on seasonality and not on mean rainfall values15.Attempts to cartography climate stability are not new. The only ones that offer data at a global scale are PaleoClim16, a free-access database of paleoclimate layers at 2.5 arc-min (~5 km) grid resolution that includes data much older than the LGM, we are presenting a new set of maps that are based on a new Climatic Stability Index (CSI). Given the markedly different speed of inferred climatic changes (see above), we divide the sets of maps into two: CSI-past and CSI-future. CSI-past is based on the 12 time periods  to present. CSI-future uses the average values of nine general circulation models (GCMs) along four time intervals until 2100 , the much wider time interval, the variety of measures  and the finer resolution (ca. 5 km) makes our proposal a very versatile tool with a wide array of applications. With both CSI-past and CSI-future we offer maps of climatic stability adhering to FAIR principles20 implemented in RStudio v. 1.4.1103. The scripts used for each methodological step are available at the Figshare repository21. After data download from primary sources , specifically for the CSI-future map set we performed an initial step aimed to obtain individual bioclimatic variables for each future time period for the four SSPs  was estimated as a measure of the amount of variation or dispersion along time series, from which the resulting output maps showed the places where climate conditions remained constant or variable across the temporal periods considered Fig. . The SD,Specifically, for CSI-past, as it includes several time periods with sea-level dropping below the present level  according to a given method (Pearson for the present case) and a threshold of statistic selected . The function removeCollinearity returns a list of uncorrelated variables according to the settings specified, randomly selecting just one variable from groups of correlated ones . The histograms were computed for these final CSI maps, which represent the frequency and distribution of CSI values. We presented the final CSI maps with two different colour ramp schemes with ArcGIS. The first consisted of defining equal interval breaks from 0 to 1. The second was based on defining 32 categories with different value breaks for past and future map sets according to the value frequency shown by the histogram plot, i.e. the category with the highest CSI values (no. 32) was 0.71–1 in the past map set and 0.356–1 in the future map set.The final CSI maps were obtained by summing the SD values of the variables selected and the subsequent outputs normalized (0 to 1) Figs. –4. Histo21 in format of map raster layers (.TIF format).Our data records are available through FigshareSD-based maps of individual bioclimatic variables, which contain a set of raster layers (one for each variable): 14 for the case of the past map set  and 19 for the future map sets based on median calculations. These layers contain individual measures of the SD of each variable, which could be independently used according to the user’s study purpose or combined according to the user’s preferences to generate a customized CSI.the five CSI-based maps presented in this study, corresponding to CSI-past, CSI-future SSP1-2.6, CSI-future SSP2-4.5, CSI-future SSP3-7.0, and CSI-future SSP5-8.5.Two sets of data records are stored:Data files share a common naming pattern:Past map set from individual variables and CSI, respectively:For “sd_past_<bioclimatic variable>.tif”“csi_past.tif”Future map sets from individual variables and CSI, respectively:For “sd_future_<SSP scenario>_<bioclimatic variable>.tif”“csi_future_<SSP scenario>.tif”In addition, for the CSI-past map set, we include a map showing the areas affected by sea-level rising periods (intergl_affected.tif) and the raster map used to remove regions with landmasses currently under the sea but that were once emerged , an intermediate (r > 0.8), or a less restrictive cut-off value (r > 0.9) was also checked. In Supplementary Tables r threshold tested are specified. All pairs of CSI rasters were compared through Pearson’s correlation analyses, which showed a very high r value, ranging from 0.93 to 1.00. All r values in Table r > 0.7, 0.8, 0.9). Our choice was using the SD, median, and threshold r > 0.8, to draw the CSI-based maps.To evaluate the robustness of the CSI index, we compared its performance by varying the statistics used for its calculation .Before concluding, however, it should be acknowledged that the fact of not using the same approaches for past and future climate stability assessments introduces a degree of uncertainty when considered as a whole. To obtain CSI-past maps we are using Supplementary Table 1Supplementary Table 2Supplementary Figures


Thanks to clinically newly introduced inhibitors of the mesenchymal–epithelial transition (MET) receptor tyrosine-kinase, MET-gene copy number gain/amplification (MET-GCNG/GA) and increased expression of the MET protein are considered very promising therapeutic targets in lung cancer and other malignancies. However, to which extent these MET alterations occur in malignant mesothelioma (MM) remains unclear. Thus, we investigated by well-established immunohistochemistry and fluorescence in situ hybridization methods, the frequency of these alterations in specimens from 155 consecutive MMs of different subtypes obtained from pleural or peritoneal biopsies and pleurectomies. Thirty-three benign reactive mesothelial proliferations (RMPs) were used as controls. MET-protein upregulation was observed in 35% of all MM-cases, though restricted to predominantly epithelioid MMs. We detected low-/intermediate-level MET-GCNG/GA in 22.2% of MET-overexpressing MMs (7.8% of whole MM-cohort) and no MET-GCNG/GA in the other 77.8%, suggesting other upregulating mechanisms. In contrast, 100% of RMPs exhibited no MET-upregulation or MET-GCNG/-GA. Neither MET exon 14 skipping mutations nor MET-fusions were detected as mechanisms of MET overexpression in MM using RNA next-generation sequencing. Finally, in two cohorts of 30 MM patients with or without MET overexpression (MET-positive/-negative) that were matched for several variables and received the same standard chemotherapy, the MET-positive cases showed a significantly lower response rate, but no significant difference in progression-free or overall survival. Our results imply that MET overexpression occurs in a substantial fraction of predominantly epithelioid MMs, but correlates poorly with MET-amplification status, and may impact the likelihood of response to mesothelioma standard chemotherapy. The predictive significance of MET-IHC and -FISH for possible MET-targeted therapy of MM remains to be elucidated. The transmembrane mesenchymal–epithelial transition (MET) receptor tyrosine-kinase (RTK) is the receptor for hepatocyte growth factor (HGF), is encoded by the MET proto-oncogene located on chromosome 7q21-31, and is normally expressed in several epithelial and mesenchymal cell types. MET can activate several signaling pathways, including the PI-3K/AKT, RAS-Rac/Rho, RAS-MAPK, JAK-STAT3, and phospholipase C pathways [The best example until now has been non-small-cell lung cancer (NSCLC), in which MET exon 14 skipping mutations affecting the corresponding splicing sites and MET-GCNG/GA occur in 3–5% and 1–6% of cases, respectively ,3,4,5. OMalignant mesothelioma (MM) is a very aggressive and challenging cancer type originating from mesothelial cells coating the pleura , or more rarely other serosal membranes, such as the peritoneum , pericardium or tunica vaginalis testis (<1%) . Its patWith regard to RTKs, how MET is expressed in MM and whether it could represent a target for this cancer type remains to be further elucidated. Although, preclinical investigations indicate that the HGF-MET axis may play an important role in mesothelioma-genesis  and thatAs initial assessment of MET gene expression in human MM, we looked at the expression of the MET mRNA in the public GEPIA database for gene expression profiling analysis in the different cancer types sampled for “The Cancer Genome Atlas” (TCGA) project ,19. The For this study, we immunostained formalin-fixed, paraffin-embedded (FFPE) tissue sections according to a previously established protocol utilizing the anti-MET SP44 monoclonal antibody and a MET-expression scoring system . As descConsistent with the fact that MET is a transmembrane RTK, we also observed in both diagnostic biopsies and P/D specimens that the expression of MET was predominantly in the membrane of tumor cells, though a certain positivity was also visible to a lesser extent in the cytoplasm . Hence, Accordingly, we examined a cohort of 155 unselected consecutive MM cases treated at our institution between 2015 and 2017. The demographic and pathological characteristics of the MM cases and of control cases with non-neoplastic reactive mesothelial proliferation (RMP) are described in p < 0.05, t-test). Similarly, of the 27 MMs with an immunoscore of 2+, 16 (59.3%) were EMMs and 11 (40.7%) BMMs . Interestingly, in the 15 BMMs with a score of 2+/3+, only the epithelioid component accounted for that, while sarcomatoid cells consistently exhibited a score of 1+/0  of the 155 MM specimens exhibited a MET immunoscore of 3+  RMPs showed a MET immunoscore of 1+ (example in p < 0.05), this difference suggests that MMs harbor fairly modest increases of MET-GCN as compared to other cancer types such as NSCLC, in which de novo high-level MET-amplification, often with more than 15 MET gene copies (“gene clusters”), can be detected by FISH analysis [The FISH analysis in the 155 MM samples showed that the average MET-gene copy number (MET-GCN)/tumor cell varied between 1.44 and 5.69. By matching the FISH results with the corresponding IHC data, we observed that MM samples displaying no upregulation of MET expression (immunoscore 0/1+) had a lower average MET-GCN/cell (between 1.44 and 2.99) than MMs with upregulated MET-receptor expression . Although significant . However, one of the cases eventually classified with intermediate-level MET-GCNG/GA had a borderline value of 5.69 for average MET-GCN/cell and a borderline MET/CEN7 ratio of 1.94. For each sample, we analyzed 100 tumor cell nuclei, taking into account possible zonal heterogeneity of GCN, i.e., we assessed 20 neighboring tumor cell nuclei from five random areas of homogenous distribution of MET signals, as previously described ,17. NeveFinally, no cases with low- or intermediate-level MET-GCNG/GA showed a MET immunoscore of 1+ or 0 by IHC. Accordingly, all 10 SMMs and all 33 cases of RMPs, which as mentioned above, all exhibited a MET immunoscore of 1+/0 by IHC, showed no MET-GCNG/GA.Given that, in our cohort of MM specimens, the IHC and FISH analyses displayed only a limited correlation between MET overexpression and MET-GCNG/GA, we addressed the question, whether in the specimens displaying MET overexpression without concomitant MET-GCNG/GA, the receptor overexpression could be explained by alternative alterations of the MET gene. Indeed, in NSCLC, another mechanism of MET overexpression is represented by mutations involving the juxtamembrane domain of the MET gene. These mutations cause aberrant splicing of the MET transcripts, in which exon 14 is skipped, resulting in reduced degradation of the MET receptor that consequently becomes overexpressed and can function as an oncogenic driver ,4,5. ThuThe RNA-NGS assay used in this study, in addition to MET exon 14 skipping, is designed to identify variants and fusions in the ALK, BRAF, EGFR, FGFR1-3, KRAS, MET, NRG1, NTRK1-3, RET and ROS1 genes. Thus, it is also pertinent to note that no variants or fusions affecting these oncogenic driver genes were identified in the 20 specimens (data not shown). These results are consistent with the paucity of oncogene activation reported in MM ,8,9 and Having established that a significant fraction of MMs shows overexpression of MET receptor, though only partially correlating with MET-GCNG/GA, we were interested in assessing whether this overexpression had a prognostic impact and would predict a response to standard-of-care first-line chemotherapy in MM. For this purpose, we compared two groups of 30 consecutive, previously therapy-naïve patients with EMM or BMM who had been treated with first-line platinum-pemetrexed chemotherapy at our institution during the period 2015–2017. In each group, MET-expression status and MET-GCNG/GA had been assessed by IHC and FISH on diagnostic biopsies with >50% of tumor cell content, as described above. One patient group contained MMs displaying MET overexpression (immunoscore 2+/3+ = MET-positive) with or without concomitant FISH-detected MET-GCNG/GA, while in the other group the tumor tissue was without MET overexpression (immunoscore 1+/0 and no MET-GCNG/GA = MET-negative). As indicated in p = 0.04) (p = 0.430). Similarly, we did not observe a significant difference in OS between the two groups  (p = 0.532)  to first-line platinum-pemetrexed chemotherapy (either neoadjuvant before P/D or for advanced disease), progression-free survival (PFS), overall survival (OS), and number of patients alive in the respective group. We noticed a significantly lower RR in the MET-positive group as compared to the MET-negative group, with fewer/no MET-positive patients showing partial/complete response and more patients with progressive disease as compared to MET-negative cases  . As indi = 0.04) , top, th= 0.934) , bottom.= 0.532) .In univariate analysis, female gender, stage IV, and lack of pleurectomy (P/D) showed a significantly negative impact on both PFS  and OS , whereasIn this study, based on a cohort of 155 consecutive MM patients, we used a well-established method of IHC ,16,17 toLikewise, previous immunohistochemical investigations of MET expression in MM have shown variable results attributable, at least in part, to the lack of standardized immunostaining and scoring procedures and to differences in types of specimens ,14,15. IYet, high-level MET-GCNG/GA might occur at low frequency also in MM, as a genomic analysis of 13 peritoneal MMs showed one case harboring focal high-level amplification of the MET oncogene, in addition to a structural rearrangement involving BAP1 and homozygous deletion of CDKN2A . FurtherMost knowledge on the correlation between MET-gene copies and MET signaling in cancer patients derives from investigations in lung cancer. Indeed, in untreated NSCLC patients, low-/intermediate-level MET-GCNG/GA is often accompanied by co-mutations in other oncogenic drivers and is less sensitive to MET-signaling inhibition by MET-TKIs, as opposed to NSCLCs with high-level MET-GCNG/GA, which at baseline are less likely to harbor other detectable oncogenic drivers and are associated with significantly higher RR to MET-TKIs ,16,23,24A further drawback of the studies that have analyzed MET-GCNG/GA in MM by FISH is the lack of a standardized method for determining MET-amplification ,14,15, wCompared to other studies, we have classified the MM specimens according to a FISH-scoring method that we and others had successfully applied to NSCLC ,17. ThisMET splice mutations result in MET overexpression and are reportedly mutually exclusive with MET-amplification in NSCLC ,3,4,5. HImportantly, despite the limitations of the study, including also the small size  of the two matched cohorts of MET-positive and MET-negative MM patients, our data indicate that MET-protein overexpression was associated with lower RR to standard platinum-pemetrexed chemotherapy for MM, while no significant difference in median PFS and median OS after standard chemotherapy was observed between the two groups. Moreover, a multivariate regression analysis revealed an independent significant impact on survival for female gender  and MM stage, but not for the MET status. We noticed also an apparent positive impact on survival for P/D, which, though, did not reach statistical significance, possibly due to the low number of pleurectomized patients in the analysis.Our results imply that MET overexpression occurs in a substantial fraction of predominantly epithelioid MMs, and that despite correlating poorly with MET-amplification status, it may negatively impact the RR of MM patients receiving standard chemotherapy. This may be due to MET being a driver of tumor cell proliferation/survival/invasiveness ,2, and tThe study examined a cohort of 155 consecutive MM cases treated at our institution between 2015 and 2017 with demographic and pathological characteristics, as specified in The immunostaining for membranous and cytoplasmic expression of MET-receptor was performed as previously described . BrieflyMET-protein expression was scored in a blinded manner (without knowing the FISH results) by one observer (E.S.-R.), assessing staining intensity  and the percentage of stained cells, thereby defining four diagnostic “immunoscores”, i.e., 3+ (strong intensity in ≥50% of cells); 2+ (moderate intensity in ≥50% of cells); 1+ (weak intensity in ≥50% of tumor cells); 0 (no staining or <50% of tumor cells stained), and considering 2+/3+ as indicative of MET upregulation (“MET-positive”) as opposed to no upregulation (“MET-negative”), as previously described ,17. EndoBefore staining the cohort of MM cases, we performed a smaller pilot investigation comparing the MET immunostaining of 30 independent FFPE diagnostic biopsies of MM with their corresponding patient-matched P/D specimens, in order to assess whether the neoadjuvant chemotherapy would have any effect on MET expression in P/D specimens as compared to the corresponding chemotherapy-naïve diagnostic biopsies. For this purpose, we selected from samples utilized in previous studies ,21,22 10FISH was performed with the Zyto-Light SPEC MET/CEN7 dual-color probe  that detects the MET gene and the centromeric portion of the MET-harboring chromosome 7, as we previously described . Briefly® Lung kit according to the manufacturer’s instructions . In addition to MET exon 14 skipping mutations, this method allows the detection of variants and fusions involving the ALK, BRAF, EGFR, FGFR1-3, KRAS, MET, NRG1, NTRK1-3, RET and ROS1 genes.For analysis of possible mutations causing an aberrantly spliced transcript of MET leading to exon 14 skipping and reduced degradation of the MET-receptor ,4,5, totTo assess the prognostic impact or predictive value for response to chemotherapy of MET-amplification and/or overexpression we compared two MM patient groups with and without MET aberrations treated during 2015–2017. For this purpose, MET-expression status and MET-GCNG/GA were evaluated as described above on FFPE diagnostic tissue biopsies obtained from 60 consecutive, previously untreated patients with EMM or BMM. Tumor samples from 30 of these 60 patients displayed MET overexpression by IHC (MET-positive = immunoscores 2+/3+) with or without concomitant MET-GCNG/GA detected by FISH. MM samples from the other 30 patients had an immunoscore of 1+/0 (MET-negative) and no MET-GCNG/GA. The two groups of patients were matched concerning gender, age, performance status, histologic subtypes, stage, asbestos exposure, smoking habit, and treatment  and were compared with respect to outcome, such as RR to chemotherapy, PFS, OS, and number of patients alive in the respective group.t-test (two-tailed) was used to detect significant differences in expression between two groups. Significant differences in the MET expression in three or more groups overall were detected using one-way ANOVA. p values adjusted for mass significance were obtained using Tukey–Kramer post-tests. For the analysis of IHC results, the distributions of H-scores in three groups were compared using nonparametric Kruskal–Wallis ANOVA and the Dunn post-test. For the correlation with outcome and the analysis of alive patients in the two groups of MET-positive and MET-negative patients a Chi-squared test was performed. OS was defined as the time from diagnosis to the time of death from any cause or last follow-up. PFS was defined as the time from diagnosis to a documented progression or death from any cause. For patients without any progression at the time of analysis, the date of last follow-up was considered right-censored. OS and PFS analysis were performed using the Kaplan–Meier method and survival curves were tested for differences by using the log-rank test. The possible independent effect upon survival of the variables considered in the study was assessed by multivariate Cox regression analysis. A p-value < 0.05 was considered statistically significant. Statistical analyses were performed using IBM Statistics SPSS software .For immunohistochemical data, the unpaired Student’s