Herein, we present a scalable approach for the synthesis of a hydrogen-bonded organic–inorganic framework via coordination-driven supramolecular chemistry, for efficient remediation of trace heavy metal ions from water. In particular, using copper as our model ion of interest and inspired by nature’s use of histidine residues within the active sites of various copper binding proteins, we design a framework featuring pendant imidazole rings and copper-chelating salicylaldoxime, known as zinc imidazole salicylaldoxime supramolecule. This material is water-stable and exhibits unprecedented adsorption kinetics, up to 50 times faster than state-of-the-art materials for selective copper ion capture from water. Furthermore, selective copper removal is achieved using this material in a pH range that was proven ineffective with previously reported metal–organic frameworks. Molecular dynamics simulations show that this supramolecule can reversibly breathe water through lattice expansion and contraction, and that water is initially transported into the lattice through hopping between hydrogen-bond sites. Heavy metals and metalloids pose major threats to health and environmental ecosystems, thus systems for low-cost remediation are needed. Here the authors report the scalable design of a hydrogen-bonded organic–inorganic framework for selective removal of trace heavy metal ions from water. Despite intensive efforts, access to safe drinking water in particular is still at risk, due in part to unacceptable levels of toxic heavy metals found in various water streams and food webs. Heavy metals and metalloids, released from geochemical cycles and anthropogenic activities, pose major threats to environmental ecosystems and can cause severe adverse health effects in humans, plants, and microorganisms. While some of these metals are essential for biological functions, they can also escape control mechanisms—such as transport, compartmentalization, and binding to designated cell constituents—via coordination chemistry and redox pathways, leading to misregulation of trafficking machinery and ion homeostasis3. Undesirable binding of heavy metals to DNA and nuclear proteins often leads to cell malfunction, oxidative deterioration of biological macromolecules, and eventually toxicity3. Effective solutions for efficient remediation of trace heavy metal ions from water with low costs are key pieces of the puzzle to promote environmental and human well-being.Rapid population growth, industrialization, and climate variability are just a few of the factors that have contributed to pushing the global ecological balance near a threshold where immediate attention is required to ensure a future of sustainability. Addressing the inextricably linked water-energy-food security nexus is critical to promote societal stability and human prosperity4. A copper imbalance can impact heart function and lipid metabolism as well as causing inflammation and resistance to chemotherapeutic drugs5. The U.S. Environmental Protection Agency has established a maximum level of copper in drinking water at 1.3 mg L−16. However, copper levels reported in drinking water in Europe, Canada, and the USA can range from ≤0.005 to >30 mg L−16. Historically, there have been several outbreaks of copper poisoning from ingestion of drinks that were contaminated with copper salts7. Given that millions of tons of copper are produced annually worldwide and it is ubiquitous across industries8, maintaining low levels of copper contaminants in water streams is a present concern. Therefore, seeking highly efficient approaches for copper removal from water is important. Beyond human health, selective separations involving copper is a relevant area of research for which traditional metallurgical solutions are limited. For example, in electrolytic nickel refining, selective removal of copper from nickel electrolysis anolytes has long been a problem plaguing the global metallurgical industry, and requires more effective solutions9.Copper, one of the most widely used heavy metals, is toxic to humans in large quantities, despite its essential role in our physiology10. Indeed, isolating and disposing of only toxic contaminants from a myriad of species coexisting in water streams—some of which are valuable catalyst materials—appears to be a costly bottleneck for the water industry. Several methods have been used to remove heavy metal ions from wastewater, including evaporation, chemical precipitation, coagulation-flocculation, photocatalysis, and membrane separation, each with advantages and disadvantages11. Adsorption has been shown to be an effective and economic approach for heavy metal ion removal, as it offers flexibility in operation, a low volume of harmful secondary products, reversibility, and is capable of producing high-quality effluent with low energy and maintenance costs10. The efficacy of adsorption methods is limited largely by the ability to design an adsorbent with the necessary physical or chemical functionality for the contaminant of choice.Challenges inherent to heavy metal ion remediation from water include incomplete ion removal, high costs, and the generation of toxic, indistinguishable ion mixtures in sludge waste streams13, polymer-based adsorbents15, layered metal sulfides17, porous aromatic frameworks4, and metal–organic frameworks20. The thioether-functionalized porous aromatic framework PAF-1-SMe was shown to be highly selective for the detection of copper ions in biological and aqueous samples4, but it was exclusively investigated as a diagnostic tool for the detection of Wilson’s disease. ZIF-8 was also recently reported to exhibit an extremely high Cu(II) adsorption capacity (up to ~800 mg g−1), establishing it among the state-of-the-art adsorbents for copper ion capture18. In general, however, most materials investigated to date for copper ion capture often suffer from one or more drawbacks, such as low selectivity, low capacity, or limited function ), and their syntheses have not been designed for affordable copper removal at scale, often using costly organic solvents or expensive preparations20. The design of selective adsorbents for efficient removal of ions with fast kinetics and high adsorption capacities, using simple and versatile synthetic routes for economic production at scale, is thus critical.A promising ion adsorbent should exhibit high adsorption kinetics, large adsorption capacities even at low contaminant levels, selectivity for the target ion, stability in the water environment, scalability, regenerability, and affordability. Several materials have been shown to be effective for copper ion adsorption, such as carbon-based adsorbents25. Such promising results motivated us to explore and exploit crystal frameworks built predominantly on the propagation of hydrogen-bonding networks ubiquitously found in sophisticated functions performed by nature26 for selective ion removal. Herein, we present, zinc imidazole salicylaldoxime supramolecule (ZIOS), a HOIF that is capable of selective copper capture without the need for postsynthetic modification and exhibits excellent separation performance relative to ZIF-8. We couple aqueous coordination chemistry and supramolecular assembly to achieve a simple and scalable synthesis of ZIOS, which is composed of trinuclear units of zinc(II), 2-methylimidazole, and salicylaldoxime. Our material exhibits unprecedentedly fast adsorption kinetics as well as high adsorption capacities and selective performance at low pH values, rendering it a promising candidate for the adsorptive removal of copper from acid mine drainage-polluted water27. In addition, ZIOS crystals are highly stable in water, exhibiting no changes in crystallinity or structure after exposure to water for 52 days. Interestingly, this stability results from the ability of the material to reversibly breathe water through lattice expansion and contraction. We envision that such supramolecular structures may inform the further development of functional crystalline structures that can be translated to macroscopic platforms for selective removal of toxic metals from water.Hydrogen-bonded organic–inorganic frameworks (HOIF) are an emerging class of adsorbents that have recently been demonstrated for selective separations due to their flexibility and the highly reversible nature of their guest uptake3)2, 2-methylimidazole (Hmim), and the copper chelator salicylaldoxime  in water around 50 °C results in the formation of a zinc imidazole salicylaldoxime supramolecule, hereafter referred to as ZIOS. The material forms rapidly as small crystals in relatively high yield (76%) following the combination of separate mixtures of H2salox/Zn(NO3)3 and H2salox/mim. Synchrotron single-crystal X-ray diffraction (SC-XRD) data 4(mim)2 4(CH3C3H2N2)2) trinuclear units, wherein H2salox and 2-methylimidazole are deprotonated and bonded directly to Zn2+ nodes through tetrahedral and pentahedral coordination by oxygen and nitrogen, respectively. Intermolecular hydrogen bonds between the hydroxyimino oxygen of H2salox and the pyrrolic nitrogen of 2-methylimidazole lead to the formation of a two-dimensional supramolecular network. The ZIOS structure is essentially nonporous, in contrast to MOFs and ZIFs28, with a Brunauer–Emmett–Teller (BET) surface area of ~12–14 m2 g−1 confirmed by both BET measurements at different activation conditions, e.g. 100 and 200 °C and grand canonical Monte Carlo simulations . Even still, we find that the Cu2+ uptake capacity of ZIOS is 40% higher than that of a sample of ZIF-8 prepared in our hands, and the supramolecular structure exhibits unprecedented rapid adsorption kinetics (via infra). This surprising performance suggests the role of a chemisorption-based uptake mechanism, based on the coordination of Cu2+ by the aldoxime and histidine-inspired imidazole pendant groups present in the host structure. Powder X-ray diffraction characterization of ZIOS revealed that the bulk, as-synthesized material is highly crystalline  weight loss at ~50 °C, likely due to traces of uncoordinated water, followed by a two-step decomposition beginning at 250 °C, as a result of ligand degradation. Differential scanning calorimetry (DSC) analysis of ZIOS  at the room temperature, and within the first 30 min both samples underwent a color change, from white to pale blue or green, respectively  images of ZIOS revealed that the material partially retains its rod-like structure following 24 h of exposure to Cu2+  to observe their adsorption kinetics, isotherms, distribution coefficient, and selective performance in complex aqueous environments containing copper in the presence of several competing ions. Details of these experiments are listed in the Supporting Information. Samples of ZIOS and ZIF-8 were initially exposed to an aqueous CuClely Fig. . This chu2+ Fig. , and eneu2+ Fig. . In otheu2+ Fig. . Crystalake Fig. .2 solution are shown in Fig. 2+ solution results in a significant drop in ion concentration to a steady state concentration <1.5 ppm in ~30 min  and is significantly larger than rates reported for other state-of-the-art copper capture adsorbents (Supplementary Table t) as a function of time shown in Fig. 30. Notably, the fast adsorption kinetics exhibited by ZIOS are accompanied by a substantial adsorption capacity: at equilibrium, the amount of adsorbed copper ions (qe) was found to be 162.9 ± 9.4 mg g−1. For ZIF-8, a smaller equilibrium adsorption capacity of 116.4 ± 5.7 mg g−1 was determined from our measurements . We note this capacity for ZIF-8 is much lower than the uptake of ~800 mg g−1 reported previously (reported surface area = 1340 m2 g−1)18, although the authors in ref. 18 determined the material uptake after only 30 min of exposure to a copper ion solution, and our data indicate that equilibrium does not occur until ~75 min. As a comparison at this 30 min mark, the uptake of our sample of ZIF-8 was 151.7 ± 4.3 mg g−1. It is worth to note that the ratio of mass concentration (in mg L−1) of Cu: ZIF-8 used in our study is ~0.17:1, whereas that reported in Zhang and coworkers’ work is 1.26:1. We also note that adsorption capacity can be sensitive to slight differences in material preparation and activation conditions.Adsorption kinetics data for ZIF-8 and ZIOS in the presence of 895 ppm CuClmin Fig. , close tn−1 Fig. . This raqsat of ~208 mg g−1 4, our material is advantageous in that it can be prepared through a much simpler synthetic route. In addition, copper uptake kinetics in ZIOS are ~30 times faster than in PAF-1-SMe4. Using the Langmuir constant, KL, obtained from single-site Langmuir model16, and Blanchard’s32 original proposed equation (see the Supporting Information), we determined that the Gibbs free energy of copper ion adsorption by ZIOS is −27.23 kJ mol−1, indicating that uptake is also thermodynamically favorable.We further equilibrated ~2.5 mg ZIOS with solutions featuring a wide range of copper concentrations . Each solution was then analyzed by inductively coupled plasma-optical emission spectrometry (ICP-OES) to determine the quantity of copper adsorbed, and the resulting data are presented in the adsorption isotherm in Fig. 2+, Na+, Ca2+, Fe2+, Fe3+, Mn2+, and Ni2+ ions, prepared from chloride and nitrate salts of these cations . Notably, even in the presence of these other ions, ZIOS adsorbed ~98.0 % of the copper(II) in solution, nearly double the amount adsorbed by ZIF-8 (52.3%), and uptake of Na+, Ca2+, Mn2+ was less than 10% in ZIOS and ZIF-8. However, we found that ZIOS also adsorbs substantial Fe2+/Fe3+ and Ni2+, namely ~93.9% and ~92.2%, respectively, compared with only ~56.1%, and ~8.3% for ZIF-8, respectively 33. Indeed, when adsorption tests were carried out using metal ion solutions prepared with pH = 2.45, copper uptake by ZIOS remained high (~97.5%) while the material adsorbed negligible iron (~14.3%) and nickel (~2.9%); in contrast, ZIF-8 adsorbed more iron (~30.0%) than copper (~14.6%)  and 4.1 (pH = 4) were reported for the commercial resin Purolite S98428 and an N-octyl-2-aminomethylpyridine-functionalized chelating resin34 under equimolar concentrations of Ni2+ and Cu2+. We further evaluated the copper selectivity of ZIOS in an ion mixture having Zn2+ as one of the contaminants and found this behavior almost unchanged  Fig. . SignifiKd, mL g−1), or the ratio of copper in the adsorbed versus the solution phase  in addition to copper(II) (see below). All peak positions were referenced to the C 1s peak of adventitious carbon at 284.5 eV. Initial deconvolution of the Cu 2p spectra for both materials  and a Cu+ complex and/or a heteronuclear Cu2+Zn2+ complex (for ZIOS-Cu), both supported by macrocyclic ligands36. For ZIF-8-Cu, pairs of Cu 2p1/2,3/2 doublets (at 952.4/932.6 eV and 954.8/934.8 eV) and the charge transfer shake-up satellites (at 962.8 and 941.8–944.8 eV) were observed, indicative of Cu+ and Cu2+, respectively. For ZIOS-Cu, the Cu 2p spectrum exhibits peaks at 952.78 and 932.98 eV characteristic of Cu+ , in addition to weaker features at 954.48 and 934.78 eV that can be ascribed to Cu2+ . The copper(I) peaks could indicate the presence of a five-coordinate Cu+ diamagnetic adduct or a four-coordinate Cu+ complex with nitrogen macrocyclic ligands, while the copper(II) peaks are indicative of a heteronuclear Cu2+Zn2+ complex37.In order to better understand the mechanisms leading to copper ion uptake in ZIOS and ZIF-8, we utilized X-ray photoelectron spectroscopy (XPS), synchrotron radiation near-edge X-ray absorption fine structure (NEXAFS) spectroscopy, and extended X-ray absorption fine structure (EXAFS) spectroscopy to characterize the materials before and after copper ion adsorption. XPS analysis of ZIOS-Cu and ZIF-8-Cu was carried out in the binding energy regions of the Cu 22+ in ZIOS results in a downshifting of the O 1s binding energies of free salicylaldoxime from 531.68 and 533.08 eV to 531.28 and 532.48 eV (HO–N and HO–C (phenol), respectively). Similar values of 531.28 and 532.38 eV (HO–N and HO–C (phenol), respectively) are observed following copper adsorption in ZIOS. In general, coordination to a metal ion results in a shifting of binding energies to lower values as a result of the electron donating/accepting ability of the metal ions38. For ZIOS, the 0.1 eV-downshift of the binding energy of the phenolic oxygen O 1s peak, along with the enhanced intensity of this peak after copper adsorption, show that this site plays an active role in scavenging copper ions. Moreover, the N 1s binding energy of ZIOS exhibits a downshift from 398.88 to 398.78 eV at the pyridinic nitrogen following copper adsorption and, interestingly, a significant increase in intensity of the peak at ~400.18 eV for C=N(–O) and/or a pyrrolic nitrogen /I(C–N) from ~0.65 for ZIOS to ~1.54 for ZIOS-Cu indicates significant translational/rotational movement of C=N(–O) and/or pyrrolic nitrogen bonding from the bulk to the surface sites at which copper ions are coordinated. It remains unclear what imidazole nitrogen participates in copper coordination. Given the tautomeric property of the ionizable imidazole ring, both are possible if the Zn-imidazole bond was cleaved at the pyridinic nitrogen by copper ions. Note that the pH value of the stock copper chloride solution is ~5.5.In ZIOS, it is possible that copper chelates with salicylaldoxime by partially transmetalating with zinc nodes originally present in ZIOS and/or with 2-methylimidazole by coordinating with the free pyrrolic nitrogen. As seen in Fig. s→1π* transition. Both features are lower in energy than the corresponding 1s→1π* transition of the pyridinic nitrogen in the imidazole ring of the ZIF-8 structure (~400.5 eV). As the imino group typically shows a transition at 398–399 eV40, the peak at 399 eV for ZIOS and ZIOS-Cu likely stems from the N 1s→1π* transition of the oximate nitrogen (C=N–O) or the imidazolate pyridinic nitrogen41. After copper(II) ion adsorption in ZIOS, this peak for the 1s→1π* transition intensifies significantly compared with the broad peaks at 403.2, 406.3, and 410.5 eV, which correspond to the ionization of aromatic N–H and transitions from N 1s→σ*N–H and σ*C–N, respectively42. This intensity increase suggests that the oxime hydroxyimino nitrogen and/or the imidazolate pyridinic nitrogen play a critical role in interacting with copper ions during adsorption. Given the close atomic radii of Zn (134 pm) and Cu (128 pm), along with the higher stability of complexes formed by copper(II) than zinc(II)43, there could be a transmetalation event where Cu2+ partially ion-exchanges with some of the Zn2+ nodes in ZIOS. During this event, the zinc–oxime bonds may be partially cleaved, leaving oxime ligands poised to scavenge copper ions. On the other hand, the unbound Lewis basic pyrrolic nitrogen atoms of the imidazole side chains in ZIOS should have a high affinity for Lewis acidic Cu2+ ions. These sites could actively coordinate with copper to form a CuZn-based heteropolynuclear complex, reactivity that could potentially mimic that found in Cu–Zn superoxide dismutase, an antioxidant enzyme found in almost all cells26 of the human body.The roles of oxime and imidazole in coordinating with copper ions in ZIOS were further elucidated with NEXAFS spectroscopy. At the N K-edge, both ZIOS and ZIOS-Cu exhibit a sharp peak around 399 eV corresponding to a 12+ adsorption does not significantly change the local environment around the Zn2+ node in the ZIOS structure. This result is indicative of negligible adsorption of Cu2+ at sites already occupied by Zn2+ and preferential coordination at oxime and/or imidazole ligands. In contrast, a stark difference in the Zn K-edge data for ZIF-8 and ZIF-8-Cu  and ~8985 and 8987 eV, respectively , which confirm the presence of Cu2+ in the samples46. However, a more detailed examination reveals some modest differences related to the peak intensities and sharpness between the two samples. The Cu L-edge NEXAFS profiles of the two samples , CuCl2 is prone to fully dissociate in the aqueous solution, rendering the inner hydration shell of copper with water molecules and nitrate anions. ZIF-8-Cu shows another predominant peak at ~2.85 Å which is within range of reported Cu–C bonds45. Furthermore, there is no evidence of Cu–N bonding in ZIF-8-Cu, supporting the fact that the parent ZIF-8 structure featuring metal–imidazole bonding is completely transformed following copper uptake. On the other hand, peaks at ~2.01 and 2.41 Å for ZIOS-Cu (averaged from different scans) align well with reported Cu–Nequatorial and Cu–Oaxial distances54, respectively, in a pentacoordinated complex with a square pyramidal geometry, suggesting some degree of chemisorption. Other peaks observed in the range of 3.2–3.9 Å for ZIOS-Cu correspond to Cu–C bonds47. From these analyses, we infer that copper predominantly chemisorbs in ZIOS by coordinating with the oxime and/or imidazole ligands, while copper ions in ZIF-8-Cu appear to be weakly bound through physisorptive interactions and also perhaps weakly coordinated within the structure. The extent to which oxime and imidazole moieties coordinate with copper in ZIOS-Cu remains undetermined.At the Cu K-edge Fig. , both ZI-Cu Fig. , inset w-Cu Fig. . The fir2 solution at pH 5.5. These results indicate that a large amount of Zn2+ still remains in ZIF-8-Cu, in spite of the structural collapse observed via SEM and PXRD, indicating that ion exchange is not the main mechanism governing copper ion uptake in this material. Rather, our data indicate that copper adsorption in ZIF-8 leads to collapse of the ZIF-8 parent structure and the formation of a new phase that has retained the majority of the zinc ions as well as incorporated physisorbed and also possibly some chemically bound copper ions. In contrast, for ZIOS, a fraction of the Zn2+ ions appear to be replaced with copper ions, leading to a structure featuring Cu2+/Cu+ coordination pockets near the coordinated Zn2+ ions in ZIOS.From ICP-OES analysis of acid-digested solid samples, we were able to estimate that our samples of ZIF-8, ZIF-8-Cu, ZIOS, and ZIOS-Cu contain ~23.4, 19.9, 18.0, and 10.1 wt% Zn, respectively. Moreover, the ratios of Cu:Zn (by mass) in ZIF-8-Cu and ZIOS-Cu are 0.9:1 and 1.7:1, respectively. In these tests, ZIF-8-Cu and ZIOS-Cu were analyzed after soaking ZIF-8 and ZIOS for 24 h in a ~895 ppm CuCl2+ nodes of the trinuclear units, chelating with salicylaldoxime ligands and leading to a partial transformation of the crystalline structure. Adsorption evidently occurs quickly enough such that ZIOS does not have enough time to further interact with water and collapse. This behavior of ZIOS aligns well with the guest-binding property of metallohosts, structures that contain one or more transition metal atoms56. This property is regulated by metal coordination or redox reactions leading to site-selective transmetalation and ultimately to the construction of heteropolynuclear supramolecular structures. The transmetalation is evident by the amount of zinc ion transferred from ZIOS into the supernatant during copper adsorption quantified by ICP-OES -containing aqueous environment, the framework expands to some extent by gradually inviting water molecules to “assimilate” themselves into the framework through the propagation of a hydrogen-bonding network. This process may then open up channels that can accept copper ions. In addition to spontaneously coordinating with imidazole pendants at Lewis basic pyrrolic nitrogen sites, copper ions may partially exchange with the Znx, Ly, and Lz  simulations were performed using the LAMMPS package with standard three-dimensional periodic-boundary conditions. Further details are listed in the Experimental. Our results show that immersion of ZIOS unit cells in water leads to an expansion of the crystal lattice and the formation of nanochannels that can accommodate water molecules through the propagation of a hydrogen-bonding network. Water incorporation occurs quite rapidly—a snapshot at 15 ns Fig.  shows a  Lz Fig. . When wa Lz Fig. , indicat Lz Fig. . A slighg(r) , and a moderately high copper ion adsorption capacity . Furthermore, selective copper removal is possible with ZIOS at pH <3, a value inaccessible with previously reported metal–organic frameworks for copper adsorption. Unlike a majority of metal–organic frameworks, our hydrogen-bonded framework is also stable to water exposure for several weeks. MD simulations show that this framework reversibly breathes in the presence of water through lattice expansion and contraction. Increasing the concentration of water in the unit cell further results in a transition from hopping to diffusion-based transport. Future efforts will investigate the regenerability of our material through copper adsorption–desorption cycling. More broadly, this study is a valuable starting point to guide further development of novel organic–inorganic hybrids that can be effectively leveraged to address at-scale removal of toxic metals from water.−1) were used in the synthesis of ZIF-8/ZIOS and ion solution preparations, respectively.Zinc(II) nitrate hexahydrate (≥98%), 2-methylimidazole (>99%), salicylaldoxime, copper(II) chloride dehydrate, calcium(II) nitrate tetrahydrate, iron(II) chloride tetrahydrate, iron(III) chloride, nickel(II) nitrate hexahydrate, manganese(II) chloride, and sodium chloride were purchased from Sigma-Aldrich and used as received. Anhydrous methanol  and Milli-Q water 2‧6H2O  and 2-methylimidazole  were separately dissolved in methanol (20 mL) and then mixed together under ambient conditions with vigorous stirring for 4 h. Solid ZIF-8 was then separated from the suspension by centrifuging for 5 min at 8000 rpm and removal of the reaction solvent. The resulting solid was then rinsed with methanol (20 mL), centrifuged, and again separated from the solvent. This rinsing process was repeated twice more, and the final product was dried in a vacuum oven at the room temperature for 18 h.ZIF-8 crystals were prepared following the procedure reported by Song and coworkers3)2‧6H2O , 2-methylimidazole  and salicylaldoxime  for 2 h at 53–57 °C and atmospheric pressure. ZIOS was then separated from the reaction suspension by centrifuging for 5 min at 8000 rpm, rinsed with deionized water (25 mL), centrifuged, and reisolated; this rinsing was repeated two times. The resulting solid was finally rinsed once with methanol (20 mL) and isolated via centrifuge under the same conditions. The final product was then isolated by decanting the methanol and dried in a vacuum oven at the room temperature for 18 h. Single-crystal X-ray diffraction confirmed that the structure of ZIOS contains Zn3(L1)4(L2)2 trimers, where L1 is salicylaldoxime and L2 is 2-methylimidazole. (Supplementary Tables 1–3) and XPS confirmed elemental composition  after degassing samples at 100 °C for 20 h. Chemical and electronic states of elemental compositions of MOFs before and after Cu2+ adsorption were evaluated with XPS collected with a monochromated Al Kα source. All peak positions were referenced to the C 1s peak at 284.5 eV. Synchrotron soft- and hard- X-ray absorption  were conducted to understand the local environment surrounding Zn and Cu metal nodes to provide insights into adsorption mechanisms. ICP-OES test was used to quantify the total trace amount of ions in the supernatant after a given time of adsorption.Further details are reported in the supporting information. Briefly, after synthesized, ZIF-8 and ZIOS were grinded with a mortar and pestle before characterization and adsorption tests. The crystal structure of ZIOS was elucidated with synchrotron SC-XRD collected on beamline 12.2.1 at the Advanced Light Source using a Bruker D8 diffractometer with a PHOTONII CPAD detector, with λ = 0.7288 Å, equipped with an Oxford Cryosystems Cryostream 800 plus, at 100 K. Representative morphology of MOFs before and after Cukd as well as the Gibbs free energy of copper ion adsorption on ZIOS. Furthermore, these MOFs were challenged with a selectivity test in which solutions having several cations and anions and adjusted pH were used. Details of experiments and calculations are listed in the supporting information.ZIF-8  and ZIOS were evaluated with different adsorption tests from which experimental data were fitted accordingly to extract adsorption performance and behavior of MOFs. In essence, adsorption kinetics test was conducted where results were fitted with a pseudo-second-order model to determine the rate of adsorption and the amount of copper ion adsorbed on MOFs at equilibrium. Adsorption isotherm was then obtained by challenging ZIOS with solutions having copper concentration values covered a wide range from 0.45 to 450 ppm for 24 h. Results were then fitted with single-site and dual-site Langmuir models. Results from this study was also used to determine the distribution (or partition) coefficient U were used in the MD simulation of this system.All-atom AMBER force fields for potential energy 63. The force field parameters of atomistic oxidized salicylaldoxime and methylimidazole were developed in previous work65 and summarized in Supplementary Tables 66 was employed for water molecules, which depicts well the dynamic and structural properties of liquid water. The VdW interaction parameters between unlike atoms were obtained by the Lorentz–Berthelot combining rule. The nonbonded interactions separated by exactly three consecutive bonds (1–4 interactions) were reduced by related scaling factors68, which were optimized as 0.50 for VdW interactions and 0.83 for electrostatic interactions, respectively. Electronegativity equalization method69 was used to calculate the atomic charges by using Multiwfn70. The schematic molecular structures and partial charges of the components for the ZIOS unit and SPC/E water molecule are presented in Supplementary Fig. In Eq. , the firSupplementary InformationPeer Review File


In-bed cycling is a novel modality for the initiation of early mobilization in the intensive care unit. No study has investigated its use in the critically ill, off-track post cardiac surgery population. Before conducting an effectiveness trial, feasibility data are needed. The aim of this study was to determine the feasibility of in-bed cycling in a population of off-track cardiac surgery patients.We conducted a prospective feasibility study in a 16-bed adult cardiac surgery intensive care unit in Ontario, Canada. Previously ambulatory adults (≥ 18 years) who were mechanically ventilated for ≥ 72 h were enrolled within 3 to 7 days post cardiac surgery. Twenty minutes of in-bed cycling was delivered by ICU physiotherapists 5 days/week. The primary outcome, feasibility, was the percent of patient-cycling sessions that occurred when cycling was appropriate. The secondary outcome was cycling safety, measured as cycling discontinuation due to predetermined adverse events.We screened 2074 patients, 29 met eligibility criteria, and 23 (92%) consented. Patients were male (78.26%) with a median [IQR] age of 76 [11] years, underwent isolated coronary bypass (39.1%), and had a median EuroScore II of 5.4 [7.8]. The mean (SD) time post-surgery to start of cycling was 5.9 (1.4) days. Patients were cycled on 80.5% (136/169) of eligible days, with limited physiotherapy staffing accounting for 48.5% of the missed patient-cycling sessions. During 136 sessions of cycling, 3 adverse events occurred in 3 individual patients. The incidence of an adverse event was 2.2 per 100 patient-cycling sessions .In-bed cycling with critically ill cardiac surgery patients is feasible with adequate physiotherapy staffing and appears to be safe. Future studies are needed to determine the effectiveness of this intervention in a larger sample.NCT02976415). Registered November 29, 2016.This trial was registered with Clinicaltrials.gov (The online version contains supplementary material available at 10.1186/s40814-020-00760-5. What uncertainties existed regarding the feasibility?What are the key feasibility findings?It is feasible to enroll critically ill, off-track patients post cardiac surgery although stringent inclusion criteria limited our sample size.What are the implications of the feasibility findings for the design of the main study?Future studies involving critically ill, off-track patients post cardiac surgery should consider broader, less strict inclusion criteria, including patients who are extubated, those who failed extubation, and those who were readmitted to the ICU from the rehabilitation ward.It was not known whether it was feasible to enroll a population of critically ill, off-track patients post cardiac surgery, how easy it would be to identify potential patients and gain consent, and whether they would tolerate this rehabilitation modality.The number of cardiac surgeries has decreased significantly over the past decade, precipitated primarily by improvements in the use of percutaneous coronary interventions (PCI). As a result of increasing PCI use, patients who qualify for cardiac surgery are older with more co-morbidities . The devEarly mobilization of patients receiving intensive care unit (ICU)-level care may reduce the iatrogenic effects of critical care , 4. EarlThe benefits of in-bed cycling in critically ill patients –8 are coBetween August 28, 2017, and March 29, 2019, we conducted a single-center pilot, prospective, feasibility study in a 16-bed adult cardiac surgery ICU in Hamilton, Ontario, Canada. Our study was approved by the Hamilton Integrated Research Ethics Board (project number 1999) and was registered with Clinicaltrials.gov (NCT02976415). Inclusion criteria were (1) cardiac surgery patients ≥ 18 years old, (2) ICU stay for > 3 but < 7 days , (3) mecCycling was performed using the RT300 Supine Cycle, a portable in-bed device which provides passive cycling, active cycling, or a combination of both with the patient in a supine or semi-recumbent position . ThroughCritical care physiotherapists delivered 20 min of in-bed cycling, Monday to Friday, to patients as part of their post-operative care. We chose 20 min for consistency with previous cycling research that included patients’ post-cardiac surgery , 9 and mWe recorded the route of oxygen delivery, mechanical ventilation settings, presence of dialysis ), agitation, and delirium as assessed by the Richmond Agitation-Sedation Scale (RASS) , ConfusiThe primary outcome was feasibility. The feasibility outcomes were 1) the ability to implement in-bed cycling into daily physiotherapy practice at least 80% of the time that patients had no cycling exemptions  sustained hypertension for ≥ 2 min ), (2) sustained hypotension for ≥ 2 min ), (3) cardiorespiratory arrest, (4) oxygen desaturation less than 88% for ≥ 2 min, (5) removal of any lines or tubes, (6) cardiac arrhythmias , (7) saphenous vein graft incision site dehiscence confirmed by the ICU intensivist, and (8) sustained increased agitation for ≥ 2 min (RASS score > + 2) .Other secondary outcomes included consent rate, hospital all-cause mortality, ICU length of stay, 28-day mortality, EuroScore II, New York Heart Association Functional Classification scores, handgrip strength, Functional Status Score for the Intensive Care Unit (FSS-ICU), 2-min walk test (2MWT), Clinical Frailty Scale, and the number of cycling sessions per patient.As no previous safety and feasibility study has been conducted in the cardiac surgery population, we based our enrollment rates on a previous safety and feasibility study of in-bed cycling in a similar-sized medical-surgical ICU . The likDescriptive statistics were used to analyze participant demographics  and baseline data . The feasibility outcome was the percent of cycling sessions that occurred compared to the number of eligible cycling opportunities. The safety outcome was the percentage of cycling sessions terminated prematurely due to the development of one of the 8 a priori adverse events. We calculated means and standard deviations (SD) and medians and interquartile ranges [IQR] for normally and non-normally distributed data respectfully. Incidence of adverse events was reported as number of events per 100 patient-cycling sessions with 95% confidence intervals (CI) . AnalysiProgression criteria are defined as criteria that inform the decision to progress from a pilot study to a larger definitive trial . MeetingWe screened 2074 patients admitted to the ICU over the 19-month period from August 28, 2017, to March 29, 2019  time from ICU admission post cardiac surgery to the initiation of cycling was 5.9 (1.4) days. Overall, our patients cycled a median [IQR] of 4  sessions which yielded 136 patient-cycling sessions and completed a mean (SD) of 113.7 (90.7) min of cycling per patient. Of the 136 patient-cycling sessions, 104 (76.5%) occurred while participants were mechanically ventilated via an endotracheal tube, 17 (12.5%) with a pulmonary artery catheter, and 14 (10.3%) during dialysis (8/136 CRRT (5.9%), 6/136 hemodialysis (4.4%)). Participants cycled during receipt of inotropes/vasopressors , benzodiazepines , and propofol . The median [IQR] RASS score pre-cycling was − 1 [1.75]. The 28-day mortality rate was 34.8%.For patients enrolled in the study, 150 routine physiotherapy sessions were also conducted. These sessions occurred separately from cycling sessions. The most common routine interventions were airway clearance techniques, including manual percussions, vibrations, and tracheal suctioning , followed by passive range of motion exercises  and sitting at the edge of the bed .Outcome measure results are available in Additional File There were 169 eligible cycling days with cycling being delivered on 136 of those days (80.5%). Of the 33 days that cycling was not delivered, 16 (48.5%) were due to physiotherapy staffing shortages  and 6 (18.2%) occurred by participant discharge from ICU without advanced notification, preventing rescheduling of the cycling sessions . Two patients developed hypotension while cycling. One patient became agitated with a RASS of greater than + 2. All 3 events lead to cycling termination with no further medical management required. Patients were able to cycle on subsequent days with no further events. The EuroScore II scores for these three patients were 1.58 (developed hypotension), 1.84 (developed agitation), and 3.71 (developed hypotension) respectively. These EuroScore II scores indicated low risk of post-operative mortality. We examined the saphenous vein graft incision sites before, during, and after cycling. No wound dehiscence was noted during our study period as determined by the intensivist on staff.During routine physiotherapy, 5 adverse events occurred in 5 individual patients : 3 oxygen desaturations to < 88% for > 2 min despite attempts to improve oxygenation and 2 developments of cardiac arrhythmias  from previous normal sinus rhythm. No further medical management beyond terminating the physiotherapy intervention was required.In the 136 patient-cycling sessions, participants cycled a total of 164.6 kilometers (km) with the median [IQR] distance cycled per participant of 5.07 [7.28] km and per session was 1.23 [0] km. The proportion of active versus passive cycling during a single patient-cycling session varied based on each patient’s status and available effort given at the time of cycling. Active cycling was noted in 45/136 patient-cycling sessions (33.1%). The minimum and maximum distances cycled during an individual session ranged from 0.49 to 2.04 km. The maximum total cumulative distance cycled by a single patient was 19.51 km. The majority  of the patient-cycling sessions were completed to the full protocol with the median [IQR] length of each patient-cycling session being 20.0 [1.3] min. The shortest cycling session was 7.9 min. The median [IQR] time to set up and complete cycling was 34 [5.7] min.For our enrollment feasibility, we enrolled 92% of our qualifying patients, which exceeded our target by 42%. Similarly, for our cycling feasibility, we were able to offer cycling on 80.5% of eligible cycling days, 10.5% above our target. This would correspond with the green traffic light color, as per Avery et al. .Our study builds on the body of knowledge investigating the safety and feasibility of in-bed cycling with critically ill patients , 11, 20 We obtained a high consent rate of 92% considering the acuity of our population. We screened 2074 patients; however, due to our narrow inclusion criteria, only 29 patients qualified. While our inclusion criteria allowed us to include patients with high acuity, it limited the enrollment of other patients who may have benefited from early in-bed cycling, such as those who required reintubation within 48 h of extubation, extubated patients who required prolonged hemodynamic support, and patients readmitted to ICU from the rehabilitation ward. While we exceeded our progression criteria with respect to recruitment of eligible patients, in consideration of future interventional trials in this population, the combination of our strict inclusion criteria and a high post-operative mortality rate would have limited the ability to assess the effectiveness of in-bed cycling on patients’ post-operative strength and functional abilities as well as their long-term functional outcomes. Future effectiveness trials of in-bed cycling in this population should consider broader inclusion criteria to prevent the exclusion of patients who may benefit from the early initiation of this modality, to ensure enrollment targets are met, and to increase the likelihood of assessing long-term functional outcomes.Feasibility was dependent upon adequate physiotherapy staffing. “Adequate” staffing for critical care physiotherapists is not defined in the literature. However, staffing limitations have been identified as a common structural barrier that can have a significant impact on patient care –26. In aAll patients were cycled on the day of their study enrollment with the mean (SD) time to initiation of cycling from ICU admission of 5.9 (1.4) days. This is more conservative than previous cycling literature , 9. TimeMore than half our study patients (56.5%) had New York Heart Association Functional Classification scores greater than 3, signifying moderate to severe symptoms of congestive heart failure prior to surgery, and 78.2% of our sample scored between class I and II on the Canadian Cardiovascular Society Scores, indicating that symptoms of angina were present only during strenuous or moderate exertion. Over 80% of our sample were identified as either “Well” or “Managing Well” on the Clinical Frailty Scale , 29. DesIn comparison, patients identified as “on-track” post cardiac surgery tend to be younger in age, male, have intact left ventricle ejection fraction, no diagnosis of diabetes, and no previous history of either congestive heart failure or cardiac surgery . The majOur protocol of 20 min of in-bed cycling was consistent with previous cycling literature in which patients post cardiac surgery were enrolled , 9. EijsAdmission to ICU is not benign. The strength and functional impairments acquired during extended stays may remain well beyond hospital discharge. Physiotherapy may mitigate the effects of prolonged critical care stays , 31, 32.There are several presumed mechanisms on how muscles may respond to physical stress during the course of a critical illness, including the ability of mechanical signals to induce protein synthesis . If muscIn this study, we demonstrated that in-bed cycling was potentially feasible and safe to implement with off-track critically ill patients post cardiac surgery. Future studies can build upon this safety and feasibility evaluation and previous effectiveness studies to determine how best to challenge critically ill patients post cardiac surgery in an effort to moderate the effects of prolonged ICU stays.This study is novel in that it is the first study to investigate the feasibility of in-bed cycling with a population of solely acutely critically ill, off-track cardiac surgery patients. We obtained a consent rate of 92% across our 19-month enrollment period. While our patients had high median pre-operative EuroScore II results, our findings suggest that in-bed cycling can be implemented with some of the most critically ill patients in the cardiac ICU.Our study had limitations. We conducted a single-center study; thus, our results may not represent the broader critically ill cardiac surgery population. We did not meet our target due to our strict inclusion criteria. Future studies should incorporate broader inclusion criteria and target subgroups of the critically ill post cardiac surgery population missed by our strict criteria.In-bed cycling can be safely implemented in a population of off-track, critically ill patients post cardiac surgery and can be feasibly conducted by critical care physiotherapists with adequate staffing. The results of this single-center feasibility study can be the basis for future studies evaluating the effectiveness of this intervention in this high-risk group. The low adverse event rate can provide confidence to broaden the inclusion criteria to include and enroll patients with or without mechanical ventilation who continue to require critical care interventions in future research.Additional file 1. Outcome Measure Results.


Patients with hemianopic field defects (HFD) might benefit from reading text in vertical orientation if they place the text in the seeing hemifield along the vertical midline.We assigned 21 patients with HFD randomly to either vertical or horizontal reading training. They trained reading single lines of texts from a computer screen at home for 2 × 30 min/day, 5 days/week, for 4 weeks. The main outcome variable was reading speed (RS) during reading standardized paragraphs of printed text (IReST) aloud. RS was assessed before training (T1), directly after training (T2) and 4 weeks later (T3). Quality of life (QoL) was assessed by Impact of Visual Impairment (IVI) questionnaire.Vertical training improved RS in the vertical direction significantly. Only patients with right HFD benefited. Horizontal training improved RS in horizontal diection significantly, but much more in patients with left than in those with right HFD. Both effects remained stable at T3. RS during training at the computer improved highly significantly and correlated strongly with RS of printed text (Pearson r= > 0.9). QoL: Vertical training showed a statistically significant improvement in the complete IVI-score, patients with right HFD in the emotional IVI-score.The improvements of RS were specific for the training. The stable effect indicates that the patients can apply the newly learned strategies to everyday life. The side of the HFD plays an essential role: Left-HFD patients benefitted from horizontal training, right-HFD patients from vertical training. However, the vertical RS did not reach the level of horizontal RS.The study was registered in the German Clinical Trials register (DRKS-ID: DRKS00018843). If data were normally distributed, parametric confirmatory statistical analyses were applied . In all other cases, we used non-parametric methods. Descriptive data were presented as medians with their interquartile range (IQR). We also used the Wilcoxon signed-rank test, Friedman test, and the Mann-Whitney U test for independent samples. With 3 test points and a significant Friedman test result, we did not perform a Bonferroni correction because it is not indicated (“closed testing procedure” ).https://libguides.library.kent.edu/SPSS/Explore).The required level of significance α was set to 0.05 (two-sided) in all statistical tests. Unless otherwise stated, given a sample size of 21, the Shapiro-Wilk normality test and graphical Q-Q plots were used to determine the shapes of distributions. All calculations were performed with listwise exclusion  and side of the HFD (left vs. right).Median individual changes (delta RS between T1 and T2) of the subgroups were calculated and the individual changes of each patient are presented as scatter-plots.The statistical values are shown in Table IReading speed (RS) during reading printed text (IReST)Reading in vertical orientationn = 11). The median vertical RS improved by 14 wpm from T1  to T2 (104 wpm) and remained stable at T3 (104 wpm)  was only 6.3 wpm (10%). Only 3/11 patients improved by at least 10 wpm , there was no significant change: T1: 70 wpm; T2: 77 wpm; T3: 73 wpm. The patients with right HFD (n = 6) yielded a statistically significant and clinically relevant improvement by 11 wpm from T1 (94 wpm) to T2  (delta RS T1-T2) shows a wide overlap between right (red dots) and left HFD (blue dots). Only 3 patients  improved by more than 10 wpm , there was statistically significant improvement of RS  from T1 to T2, which remained stable at T3.For the total cohort  did not show any statistically significant changes during horizontal reading.Training group V (n = 10) showed a statistically significant increase of RS of 14.9 wpm from T1 (112.50 wpm) to T2 (127.40 wpm), of 12 wpm from T1 to T3 (124.50 wpm), and no change from T2 to T3, i.e. the training effect remained stable.Training group H  (delta RS T1-T2) was 11 wpm for group V and 12.5 wpm for group H.The individual change of RS during the training period showed a wide overlap between the groups without a clear advantage for the horizontal training group :HFD Fig. :Fig. 5Ren = 21) into groups according to the side of the HFD (independent of the training group), patients with left HFD (n = 11) showed a statistically significant increase of median RS during horizontal reading from T1 (113.0 wpm) to T2 (129.8 wpm), see Fig. Separating all patients  was 15.1 wpm (22.2%) in left HFD and 5.6 wpm [8.8%] in right HFD (delta RS T1-T3).The individual change RS correlated highly with cRS (RS during the training) (see below). No correlations were found between RS at T1 and the change of RS (from T1 to T2), regarding age, disease duration, and size of macular sparing.n = 11) showed a statistically significant improvement of the reading speed at the computer during the training (cRS) between the median of the first 3 days (72.93 wpm) and the median of the last 3 days (103.35 wpm) of the vertical reading training (see Table Group V ( 3 days 7.93 wpm an = 10) improved cRS during the horizontal reading training, but this change was not statistically significant: the mean of the first 3 days of training was 126.72 wpm; the mean of the last 3 days of training was 148.70 wpm. (see Table Group H (III.Training intensitycRS (during the home training) correlated highly with RS (at testing in the clinic with the printed IReST charts) see Fig. .III.Train = 21) was 1236.45 min  (103%). As the actual average training time per day for each patient was 62 min, this means that there was good compliance. There was neither a statistically significant correlation between training intensity and change of RS (T1-T2), nor between training intensity and change of cRS during the training at home .IV.QuestionnairesQuality of Life (IVI)The study design requested that each patient should train for 4 weeks, 5 days per week, 2 × 30 min per day (1200 min = 20 h). The actually measured median cumulative training time of all patients , we found a statistically significant improvement in the functional IVI-score from T1 (1.45) to T2 (1.05), which remained stable at T3 (1.05). The IVI reading-score showed an improvement from T1 (1.5) to T2 (1.0) and from T1 to T3 (1.00), which was statistically marginally significant. The complete IVI-score showed statistically significant improvement from T1 (1.25) to T2 (1.00) and from T1 to T3 (1.11) indicating that the effect remained stable after training. An improvement of the complete IVI-score was found in 11/12 patients who improved their RS by more than 10 wpm, but was found by only 3 patients who had not improved their RS. This indicates that a successful training was considered beneficial in everyday life by the patients.When all patients were analyzed together  to T2 (1.00), group H did not show any significant improvement.When the total cohort was separated by side of HFD, in left HFD, no significant improvement was observed. In right HFD, the improvement was significant in the emotional IVI-score from T1  to T3  . The reading IVI score tended to improve: T1: 1.50 (IQR 1.00 to 2.00); T2: 1.0 (IQR 0.50 to 1.50); T3: 1.00 (IQR 0.50 to 1.00), p = 0.07, Friedman’s test. Post-hoc tests were not applied, as Friedman’s test did not show a significant improvement.b)Specific questions regarding reading performancec)Feedback regarding the training programSome patients reported spontaneously that reading was „fun again“, that they were reading “much more than before”, and that they were reading “more comfortably”. The questionnaire showed that the amount of reading in daily life increased in 6 patients, the strain decreased in 5 patients. This was assessed at T1, T2 and T3.As not all patients answered all the questions, we can only report the absolute numbers. The questions were asked after the training at T2.How demanding was the training for the patients: Five patients described the training as slightly demanding , 7 patients reported the training to be demanding , 4 found it exhausting . This effort decreased during the training period in 9  and remained equal in 10  patients.d)Cognitive StatusQuestion regarding the handling of the program: It was easy for 9 and needed help in another 5 patients. Four patients reported difficulties. Reading in daily life was experienced as easier in 5, equal in 13 patients and harder in none.The MoCA score was not statistically different, neither between the reading training groups nor between the groups divided by the side of the HFD, and did not change during the training period , experienced a statistically significant improvement of their median vertical RS. This effect remained stable at T3, and they improved their RS not only during the training at the computer, but also during reading the printed IReST. This means that the patients applied their improved reading performance to everyday life. The finding that only patients with right HFD benefited regarding vertical RS is in accordance with the finding by de Jong et al. . This isHorizontal training (group H) led to statistically significant improvement of RS in horizontal orientation. The effect remained stable at T3. Despite the improvement during vertical training, the vertical RS at T2 (104 wpm) did not reach the level of horizontal RS . It should be taken into account that most people have practiced horizontal reading throughout their lives, whereas vertical reading is unfamiliar and challenging, e.g. because of the perceptually unusual word shapes and the necessity to perform reading saccades in a vertical direction. In an unpublished pilot study, normally sighted subjects found that vertical reading was quite demanding and tiring. However, it should be kept in mind that normally sighted people will not have any functional advantage using a vertically orientated text, whereas HFD-patients might have this advantage by shifting the line of text into their seeing hemifield. Our results allow hypothesizing that these patients might benefit only after much longer training of reading in the vertical direction.In fact, the difference between horizontal and vertical reading performance disappears, if readers are used to read in both directions, as in Chinese and Japanese . FurtherIn our total cohort of patients (V + H), we found a statistically significant increase of RS from T1 to T2 for horizontal reading. This effect remained stable at T3. Regarding the side of the HFD, patients with left HFD showed stronger improvements from T1 to T2 than those with right HFD from T1 to T3 during horizontal reading, but not in vertical reading. Patients with right HFD have the problem of having to perform saccades into the blind hemifield, but can at least improve by reading more frequently. The patients with left HFD have the advantage of getting through the line rather easily during horizontal reading and might need some more (unspecific) training of the return sweep to the next line.Even though we found statistically significant effects in the groups and subgroups, we should consider that the changes of RS were individually very different and did not show very clear effects. One limitation of the current study is the small sample size, which makes it difficult to form subgroups. The recruitment of the patients was very difficult, which is shown in Fig. We consider a change of 10 wpm in these patients as clinically relevant. The value is based on our long-standing clinical experience and has been used in previous studies e.g. , 35). In. In35]).There are two RCTs on reading training in patients with HFD that used different approaches. Spitzyna et al., (2006)  applied The strong correlation between reading speed during home training at the computer (cRS) with reading speed of printed paragraphs (RS) in the clinic shows that the method of measuring reading speed by software allows an assessment of the training results at home. The advantage is that the training effect can be determined by software without an additional visit in the clinic. This could save additional effort for the patient and lowers the costs for the health insurance, which would allow monitoring the status of patients with reduced mobility by “telemedicine”. The strong correlation between RS during reading printed text and cRS during the home training at the computer shows that the measurement of reading speed in both conditions is reliable and shows that the data is robust. This also indicates that the patients actually read the texts during the training at home without a supervising person.Not all of the objective improvements were reflected in the subjective reports by the patients. The fact that QoL improved in group V and in right HFD is in accordance with the finding that patients with right HFD and vertical training benefited most.The cognitive status (MoCA) of our patients did not change. There was neither an increase (by more intensive reading) nor a decrease  of the cognitive status of our patients.In summary, we found significant improvements of reading speed in two groups using reading training in horizontal or vertical orientation. Training in vertical text orientation improved RS in patients with a right hemianopic field defect, but does not reach the speed attained by reading text presented in horizontal orientation.In patients with a left HFD, the vertical training did not offer an advantage, because their main problem is finding the beginning of the new line, which they can train more easily by horizontal reading.


New tools to calibrate different types of scattering experiments suitable for static and moving detectors are presented. pyFAI suite for processing scattering experiments acquired with area detectors are presented. These include a new graphical user interface for calibrating the detector position in a scattering experiment performed with a fixed large area detector, as well as a library to be used in Jupyter notebooks for calibrating the motion of a detector on a goniometer arm (or any other moving table) to perform diffraction experiments.New calibration tools in the  The calibration issue is currently worked around by providing users with the proper geometry description or even the reduced data; however, this prevents re-processing at home institutes and also re-interpretation of data, which is needed as a part of the open-data initiative (Wilkinson etc.). Using a fixed-position setup combined with the speed of modern detectors (up to a few kHz), it is easy to follow chemical reactions or other physical processes in situ. Even with fixed geometries, determining the detector position is still an issue for the majority of users. Hence, a new graphical user interface has been developed for the Python fast azimuthal integration library pyFAI  area detectors mounted on moveable goniometer arms for powder diffraction . Detectors at synchrotrons are often mounted on goniometer or translation tables to provide the degrees of freedom (hereafter DoFs) needed to align the beamline, although those DoFs are rarely used for data acquisition. One counter-example is reported by Gao  al. 2016, where tq-range in one single frame  to acquire powder diffraction or PDF data on a larger q-range with no additional cost or investment.Pair distribution function (PDF) analysis typically requires very large area detectors and higher energies to be able to cover the needed high pyFAI library  as a function of motor positions is described.The 2.pyFAI is a Python /λ. pyFAI also provides additional tools to calibrate the detector position, i.e. to determine its location in space by means of Debye–Scherrer conical rings resulting from the intersection of the diffracted X-ray beam with the detector surface. The observed rings are deformed into ellipses when the detector is planar but slightly inclined. The azimuthal integration is performed in two steps. The first is a pixel-wise transformation corresponding to the image correction,Iraw is the raw signal of the detector, Idark is the dark current image , F is the factor accounting for the flat-field correction, Ω is the solid angle subtended by a given pixel, P is the polarization correction term and A represents the detector’s apparent efficiency due to the incidence angle of the photon on the detector . Iraw may be normalized by the incoming flux I0, which is independent of the pixel position. The numerator of equation (1)r). The intensity averaged over all pixels located at a given radial value is then simply the ratio of the two histograms which bin corresponds to the radius of interest [equation (2)]ci is used to describe pixel splitting and corresponds to the fraction of the pixel area falling into the specific histogram bin (0 ≤ ci ≤ 1). The multiple pixel-splitting schemes available in pyFAI, called full-splitting, bounding-box-splitting and no-splitting, have been described by Ashiotis et al. , which used equation (4)The integration scheme presented in equation (2)3.pyFAI integration scheme of multiple images taken at various positions was first reported by Kieffer & Wright . Hence, integrated intensities in multi-geometry mode are orders of magnitude larger than usual, the scale factor being the solid angle of one pixel at the PONI, i.e. dist2/(pixel1 × pixel2). Errors are propagated in an similar way to equation (3)The ight 2013. The pro4.4.1.calibrant. The rings are extracted automatically from the image (by subtracting a smoothed version of the image) and control points are placed at the local maxima on the rings. The geometry of the experiment is obtained by refining geometric parameters using a least-square fitting of the 2θ values calculated for the different control points.The calibration of a detector position is performed using several Debye–Scherrer rings collected from a reference powder called the FIT2D . The geometry used in pyFAI is based on the definition of the PONI  on the detector plane (or the plane z = d3 = 0 when the detector is non-planar and z varies from pixel to pixel). This geometry is inspired by SPD . Some geometry conversion tools are provided by pyFAI and documented by Detlefs & Kieffer  from the sample position to the PONI. (ii) poni1 and poni2: space coordinates of the PONI (in metres) within the detector plane (z = d3 = 0) along the slow- and fast-reading dimension of the detector image . (iii) rot1, rot2 and rot3: the rotation angles (expressed in radians) of the detector placed at the proper distance from the sample, with respect to the three axes of the laboratory reference system. The detector is first rotated around the vertical axis (rot1), then around the horizontal axis (rot2) and finally around the incoming beam direction (rot3). In a more mathematical way, this givesR1 and R2 are left-handed, while R3 is right-handed, which is a legacy from previous versions of pyFAI.The geometrical parameters to be refined are the following: (i) The strength of this geometry parameterization is that it describes any detector position in space. The drawback is that some parameters are correlated or not optimized:dist-wavelength: reducing the wavelength is nearly equivalent to increasing the distance unless the diffraction angle is large (2θ > 30°). It is advisable to fix one of these two variables unless the data quality are good enough and the scattering range is very large. This is an intrinsic limitation of the single-image approach.(i) rot1-poni2 and rot2-poni1: as small rotations can be mistaken for larger translations. Fixing one of the two decreases the uncertainty of the other by several orders of magnitude. The user interface offers a ‘SAXS constrains’ button to lock the detector normal to the direct beam.(ii) rot3: cannot be refined from Debye–Scherrer cones as they are, by definition, invariant by rotation around the incoming beam. The rot3 parameter can be assessed using an anisotropic contribution such as the polarization effect or some textures in the sample. This rot3 parameter is used to describe the azimuthal rotation of the experimental setup or of the sample, for example, when calculating pole figures.(iii) azimuthal integrator object, ready to perform the azimuthal regrouping of images coming from the detector.Those calibration parameters containing the geometry of the experimental setup are saved in text files with a .poni extension, also containing the detector definition and the wavelength. Such a file can subsequently be loaded into an 4.2.pyFAI since the origin of the project  design was developed on top of (i) Experimental settings. This first tab lets the user select the wavelength (or the energy), the reference material for calibration  and choose the type of detector, either from a list of 50 provided, or by defining a new detector Fig. 2.(ii) Mask drawing. This tab allows us to mask-out unwanted regions/pixels from the image, either based on their intensity (thresholding) or simply by drawing polygons on the image. Different masks can be saved, retrieved and assembled Fig. 3.(iii) Peak picking. Individual peaks and arcs of rings can be segmented out  and assigned  to different rings, each associated with a single reflection of the calibrant Fig. 4.(iv) Geometry fitting. At this stage, the detector position and wavelength are fitted against peak positions and ring numbers. Any of the parameters can be fixed or let free for refinement, possibly with boundaries. The  positions of the beam centre and the PONI, and the expected positions of rings are overlaid onto the diffraction image to visually assess the quality of the fit. The sample, detector and direct beam can also be visualized in 3D Fig. 5.i.e. powder diffraction profile and caked-image) to further validate the modelling of the experimental setup  Integration. This tab displays the 1D and 2D integrated patterns .The calibration of the detector position on a fixed goniometer position can be performed with 5.1.pyFAI accepts an arbitrary number of motors attached to the goniometer/translation stage. Let motors be a list of n motors moved during the acquisition: .Here, the difficulty is not in the calibration, but rather in the variety of goniometers and translation stages which may be controlled by one or multiple motors. The goniometer description implemented in Calibration of the goniometer involves defining a model and optimizing it:m static parameters for the model which are used to describe the position of the detector, m being the number of DoFs of the model: params = .(i) Choose a list of transformation functionsmotors) and goniometer parameters (params) into the six PONI parameters used to initialize an azimuthal integrator in pyFAI:(ii) Define the six (iii) Optimize the parameter set of the goniometer (params) so that, for any position of the motors, the detector position is expressed as a PONI-parameters set.exp position of the peaks over all control points. Those 2θexp are compared with the theoretically expected 2θtheo values calculated from the calibrant d-spacing and the wavelength,scipy.optimize.minimize function from SciPy , but should be prevented from executing any arbitrary code (security). Moreover, those functions should be made persistable on disk (i.e. saved) and their restoration should be possible without having to re-perform the calibration, nor creating vulnerabilities.The six PONI parameters returned are then used to calculate the experimentally observed 2θNumExpr library , with a list of motor names (pos_names), parameter names (param_names), as well as a set of six formulae, one for each of the six PONI parameters.In the following example, the definition of those functions is simply the creation of an object from the 5.2.q or longer distances for better Bragg peak separation.At the ESRF, the protein crystallography beamlines use large area pixel detectors  placed on a translation table which allows collection of the data at the optimal distance: shorter sample-to-detector distances to explore the region for high-MX-calibrate tool from the pyFAI suite is available for the calibration of many images taken at various distances. This can also be interpreted in terms of the goniometer setup , where the sample-to-detector distance is modelled as a simple linear function of the table position:The pyFAI: poni1, poni2, rot1 and rot2. rot3 is forced to zero while the distance is defined as a linear function of the motor position.In the example above, a six-parameter model was chosen for the goniometer geometry, most parameters matching exactly those of Jupyter notebook  from the metadata written in the image headers, thus a set of control points is available prior to data processing. The translation table is calibrated using all control points and this geometry transformation object. As a result, all images have been integrated azimuthally. Fig. 72 calibrated using this methodology. The blue curve corresponds to this initial model refined; the two peaks look ‘doubled’, indicating a poor modelling of the geometry.The content of this tutorial is available as a dist, poni1 and poni2) have been re-fitted and all data integrated with the new model, one obtains the orange curve in Fig. 7To address this poor modelling, another transformation function is defined, with a few additional DoFs on the PONI position . Once three parameters  for powder diffraction measurements. The initial idea was to use a small region of interest (ROI) in the centre of an area detector, and check whether it was possible to rebuild a powder diffraction pattern when moving the detector at various goniometer angles. This ROI has to be of limited size in the dimension orthogonal to the ring and as extended as possible in the tangential direction, with the limit of the curvature of the ring . We considered an ROI 10 pixels wide within a Pilatus 100k (487 pixels × 195 pixels) which means using only 2% of the total width of the detector (different lines being used for different bins). By using the full detector area, the signal/noise ratio could be improved by a factor of seven [(487/10)1/2] if the movement of the detector on the goniometer is perfectly predictable and reproducible.Probably the main application of this work is to place a small area detector on a goniometer 2θ arm in replacement of a punctual detector (geometry transformation is defined with rot2 (in radians) as a linear transformation of the motor position (pos is the default motor name), here measured in degrees by the goniometer. The scale parameter, scale = π/180, is used to convert angular units. The other parameters dist, poni1, poni2 and rot1 are directly mapped to pyFAI’s parameters. As done previously, rot3 is kept fixed at zero:When the detector arm is moving in the vertical plane , a simple Jupyter notebook , most have only one ring and only a few images display two rings. Even if those images are technically suitable for static detector calibration, they still present a challenge due to the large diffraction angles (2θ > 30°), the low curvature of the rings and the difficulty in assigning the picked peaks with the proper calibrant ring number.This example is also available in a rot1 = 0) to guarantee some consistency between the calibrations.Four images have been manually calibrated, fixing the vertical rotation axis of the detector  over all images, but Fig. 10rot1. A new model was tested where both the rot1 and rot2 values are scaled with the motor position. After refinement, the cost function dropped by a factor two and the low-angle peaks became sharp  is a factor of ten worse than the highest achieved resolution with secondary monochromator on insertion devices . The average peak resolution full width at half-maximum FWHM = 0.02° obtained on LaBet al., 2003et al., 1998rot3 = π/2.A third example of goniometer calibration is available in the work by Kieffer & Blanc 2017. It corrThis generic method has even been extended to strip detectors like the Mythen detector manufactured by Dectris as shown in the tutorial on the calibration of the array of nine Mythen detectors , this shrinkage of pixel sizes naturally leads to higher quality powder diffraction patterns. Unfortunately, to keep the covered 2θ range constant, one would need to multiply the number of pixels by the square of the pixel size reduction factor, and the associated infrastructure for read-out and data transfer accordingly. Moving the detector offers a flexibility which removes this limitation but makes the experiment slightly slower – though still suitable for 7.pyFAI has been developed to ease the calibration of an experimental setup with static detectors, especially for novice users. The concept of calibration of the detector position has been extended to fit the detector position as a function of the motion of a goniometer. Once a few fixed positions of the goniometer have been calibrated, a model can be optimized and the detector position can be extrapolated at any goniometer configuration. By acquiring multiple images at various positions, these images can be integrated together to produce a high-q powder diffraction pattern of quality equivalent to that acquired with a much larger detector, opening up new opportunities for insitu experiments and PDF measurements on most synchrotron diffraction beamlines.The new graphical user interface of


Tumours with high expression of B7-H3 showed larger spatial differences of CD8+ T cells (86.4/mm2 in tumour centre vs. 414.9/mm2 in invasive front) when compared to B7-H3-low group  (p < 0.001). This study provides insight into the expression pattern of B7-H3 in GC of Western origin. In GCs with higher level of B7-H3 expression, CD8+ T cells were spatially suppressed in the tumour centre suggesting that B7-H3 might be involved in tumour escape mechanisms from the immune response.CD8+ T cells are the main effector cells of anti-cancer immune response that can be regulated by various costimulatory and coinhibitory molecules, including members of the B7 family. B7 homolog 3 (B7-H3) appears as a promising marker for immunotherapy; however, its significance in gastric cancer (GC) is unclear yet. We evaluated the spatial distribution of CD8+ T cells in relation to the expression of B7-H3 by double immunohistochemical staining. The level of B7-H3 intensity was scored manually (0–3) and dichotomized into B7-H3-low and B7-H3-high groups. The distribution and density of CD8+ T cells was analysed using whole slide digital imaging. B7-H3 was expressed mainly in the stromal compartment of GC ( The tumour immune microenvironment (TIME) can be categorized into three classes : (a) theB7 homolog 3 protein  is one of the immune checkpoint molecules of the B7 family. Although the exact receptor of B7-H3 has not been discovered yet , this mon = 48; diffuse, n = 27; mixed, n = 6; unclassified, n = 15) were included in the study. Among 73 B7-H3 positive GCs (76%), B7-H3 expression was found within the tumour stroma (n = 60) or both in stromal cells and neoplastic cells (n = 13). After assessing the intensity of B7-H3 expression, 55 cases showed negative or low staining, and 41 cases showed moderate or strong staining .The median overall survival (OS) for the entire cohort was 17.5 months, and the median cause-specific survival (CSS) was 18.4 months. No significant differences were found in OS and CSS between B7-H3-low and B7-H3-high groups (2) differed significantly between both tumour compartments  than in the invasive front (IF) (258.1 cells/mm2). Tumours with higher B7-H3 expression in the stroma showed greater spatial differences of CD8+ T cells (86.4/mm2 in TC and 414.9/mm2 in IF) when compared to the B7-H3-low group  (The median density of CD8+ T cells (cells/mm< 0.001) . Addition = 415) was significantly higher than that in normal gastric tissue  .Data from The Cancer Genome Atlas Stomach Adenocarcinoma (TCGA STAD) cohort were retrieved for quantitative validation analysis using TCGA Xena computational tool (UC Santa Cruz) . The B7-Studies have demonstrated that the analysis of tumour infiltrating lymphocytes (TIL) alone cannot explain the complex crosstalk between tumour cells and their TIME. Here, we have presented a potential relationship between B7-H3, an immune checkpoint molecule, and CD8+ T cells, the main effector cells involved in tumour surveillance. The major findings of this study are the following: (1) B7-H3 is expressed mainly within the tumour stroma; (2) B7-H3 expression correlates significantly with tumour location, Lauren phenotype, and pT category; (3) median density of CD8+ T cells differs significantly between TC and IF; (4) in cases with higher stromal B7-H3 expression, restricted CD8+ cell infiltration in the TC was observed. Taken together, our results suggest a potential B7-H3 role in the immunosuppressive TIME of GC. However, while there is no specific human receptor for B7-H3 described yet, the exact mechanism of this interaction remains unclear.Most studies of TILs (including CD8+ T cells) and B7-H3 in GC are carried out on Asian cohorts. According to their meta-analyses, higher intratumour CD8 expression is associated with an improved overall survival ,12. Distn = 24), Gullo et al. demonstrated that T cells are enriched at the invasive margin of GCs, as well as in EBV+ cases [n = 18) showed T cell exclusion and possessed CD8+ cells predominantly at the invasive margin [By analysing GCs with lymphoid stroma , B7-H3 expression in stromal cells was described. By examining the co-expression of B7-H3 with CD31 and alpha-SMA, they detected stromal B7-H3 expression in 62.7% of GC cases, mainly in cancer-associated fibroblasts, which were the major component of the tumour stroma [The association between B7-H3 positivity and CD8 expression has been reported. Guo et al. examined the expression of PD-L1, B7-H3 and B7-H4 during different stages of gastric carcinogenesis. In their set of 50 GCs, 78% cases were B7-H3 positive, and a negative correlation between B7-H3 in the tumour and the density of CD8-expressing cells was reported . Here, wIn conclusion, the results indicate that B7-H3 within GC stroma could also play a role in regulating cell-mediated immune responses against cancer in patients of Western descent. Thus, it could serve as a great target of interest for different novel cancer immunotherapeutic strategies, especially in immunologically “cold” tumours.n = 48; diffuse, n = 27; mixed, n = 6; unclassified, n = 15) were included in the study. Specimens were routinely fixed in formalin and embedded in paraffin.A total of 98 gastric adenocarcinoma cases were selected by stratified sampling out of a consecutive series of GC patients treated with primary total or partial gastrectomy from the archive of the Institute of Pathology, University Hospital Schleswig-Holstein, Kiel, Germany. The following clinicopathological characteristics were collected: gender, age at diagnosis, tumour location, tumour phenotype by Lauren, pTNM stage, histological grade (G), UICC stage, as well as survival data. All patient-related data were pseudonymized. Two cases were excluded from the cohort as there was no clear invasive margin evaluable in the slides. Finally, a total of 96 cases . In the first step, antigen retrieval was performed using BOND Epitope Retrieval Solution 2  for 20 min, followed by incubation with anti-human B7-H3 . Detection and visualization were done with the Bond Polymer Refine Detection kit . In the second staining step, antigen retrieval was performed again using BOND Epitope Retrieval Solution 2 , followed by the second antibody incubation with anti-human CD8α . Finally, detection and visualization were done with the Bond Polymer Refine Red Detection .B7-H3 expression was scored manually by two pathologists (D.U. and C.R.) for staining intensity and localization. Four representative cases were selected for the adjustment of staining intensity  and used as reference standard for the evaluation of the entire cohort. Inconsistent results were resolved by consensus review. For statistical purposes, cases were divided into B7-H3-low (score 0/1) and B7-H3-high (score 2/3) groups as proposed by a previous study .2; cell simulation with 2 µm of maximum growth as the optimal threshold for cell border generation; cell classification by double staining immunohistochemistry markers with red chromogen threshold of 0.32 and brown chromogen threshold of 3.0, respectively. These settings were used for all the images and carried out by a single author, who was blinded to patient outcomes. To avoid the false results due to different artefacts , marking of tumour compartments was done manually by using the viewer and painting program VMP. The tumour centre (TC) was defined as intratumoural area comprising cancer cells and desmoplastic tumour stroma with no direct connection to the peritumoural non-tumourous tissue. The invasive front (IF) was defined as a narrow band-like area at the tumour–host interface with a width of up to 250 µm between the invasive margin of GC and adjacent non-tumourous tissue.Digital images of tissue sections were obtained using a Leica SCN400 microscopic whole-slide scanner  at 40× nominal magnification, which corresponds to a resolution of 0.25 µm per pixel. To detect CD8+ T cells, digital image analysis with Definiens Tissue Studio  was performed. The following detection settings were used: 20× nominal magnification for digital analysis; nucleus detection with haematoxylin threshold of 0.15 and typical nucleus size of 25 µmp < 0.05 was assumed.Data were analysed using SPSS v25.0.0.2 . Differences in clinicopathological variables between B7-H3 staining intensity groups were determined using Kendall’s tau test for ordinal variables and Fischer’s exact test for non-ordinal variables. Survival data were analysed by the Kaplan–Meier method and compared using log-rank test. Differences in CD8+ T cell densities between B7-H3 groups were analysed using Wilcoxon Signed Ranks test and Mann–Whitney U test. A significance level of http://xena.ucsc.edu (accessed on 14 February 2021)) [http://timer.cistrome.org (accessed on 14 February 2021)) [Data from TCGA STAD cohort were retrieved for quantitative validation analysis using TCGA Xena computational tool ) . The effy 2021)) .


Type 1 Diabetes (T1D) is a T cell-mediated autoimmune disorder caused by the destruction of insulin-secreting cells. B7-H3 (CD276) plays a vital role in T cell response. However, B7-H3 expression and its clinical significance in T1D remain unclear. The aim of this study was to investigate the correlations between the expression of B7-H3 and clinical parameters in T1D patients. The possible role of B7-H3 gene variants with T1D was also discussed.Four B7-H3 single nucleotide polymorphisms (SNPs) were genotyped in 121 T1D patients and 120 healthy controls by polymerase chain reaction (PCR) direct sequencing. Expression of membrane B7-H3 (mB7-H3) in peripheral blood lymphocytes was determined by flow cytometry. Levels of soluble B7-H3 (sB7-H3) in serum were analyzed by enzyme linked immunosorbent assay (ELISA).P < 0.05). The concentration of sB7-H3 in serum increased in T1D patients (P < 0.0001). We also observed that B7-H3-T-A-C-T was associated with the decreased release of sB7-H3 but not the membrane form.The B7-H3 haplotype T-A-C-T was less frequently observed in T1D patients compared to the controls . B7-H3 expression on monocytes showed significant upregulation in T1D patients and was positively correlated with several clinical features including ALT, fast C-peptide 120 min, HbAlc, IFN-γ, IL-6 and TNF-α (B7-H3 may act as a potential biomarker related to the pathogenesis of T1D. The B7-H3-T-A-C-T polymorphism variant is associated with the low risk of T1D as well as less release of sB7-H3. Type 1 diabetes (T1D) is a chronic autoimmune disease that leads to the selective loss of insulin-producing β-cells by activating the T cells –6. B7-H3+ and CD8+ T cell proliferation, cytotoxic T lymphocytes (CTLs) induction and interferon-gamma (IFN-γ) secretion in vitro  range; R is the correlation coefficient between alleles.Additional file 5: Fig. S4. Expression of mB7-H3 in HC and T1D patients. Bars show the mean and SD of mB7-H3 mean fluorescence intensity (MFI).