Transport properties are one of the most crucial assets of hydrogel samples, influencing their main application potential, i.e., as drug carriers. Depending on the type of drug or the application itself, it is very important to be able to control these transport properties in an appropriate manner. This study seeks to modify these properties by adding amphiphiles, specifically lecithin. Through its self-assembly, lecithin modifies the inner structure of the hydrogel, which affects its properties, especially the transport ones. In the proposed paper, these properties are studied mainly using various probes (organic dyes) to effectively simulate drugs in simple release diffusion experiments controlled by UV-Vis spectrophotometry. Scanning electron microscopy was used to help characterize the diffusion systems. The effects of lecithin and its concentrations, as well as the effects of variously charged model drugs, were discussed. Lecithin decreases the values of the diffusion coefficient independently of the dye used and the type of crosslinking. The ability to influence transport properties is better observed in xerogel samples. The results, complementing previously published conclusions, showed that lecithin can alter a hydrogel’s structure and therefore its transport properties. Hydrogels are hydrophilic polymeric materials with a three-dimensional structure (or 3D networks of hydrophilic polymers) capable of swelling in aqueous solutions or biological fluids and also possessing unique properties, such as biocompatibility, biodegradability, superabsorbency, viscoelasticity, softness, and responsiveness to external stimuli (pH and temperature) ,2,3. TheIn the case of designing systems for the controlled release and delivery of drugs, special attention should be given to the mechanism of transportation and its correlation with the microstructure of hydrogels. The physical and mechanical properties of such systems can easily be improved (1) by adding a structure modifier; (2) by means of binary systems composed of two or more mixed polymers, such as interpenetrating polymer networks; (3) by using self-reinforced composite materials composed of fibers embedded in a matrix of the same polymer; and (4) by means of the sol-gel reaction to produce silica reinforcement . AnotherThe characterization of hydrogels with a modified internal structure is often based on the analysis of their mechanical properties as well as their biocompatibility and biodegradability. In our previous paper , we provTransport properties generally include viscosity, thermal conductivity, and the diffusion coefficient (diffusivity), which indicates the rate at which specific (per unit volume) momentum, heat, or mass are transferred . StudyinIn this article, we present a study of the transport properties of hydrogel and xerogel systems determined on the basis of the given release experiments. In general, we investigated the diffusion of organic dyes  and calculated the diffusion coefficients for hydrogel systems with different weight concentrations of lecithin as a structure modifier.According to previously published data and to extend the results obtained in the previous article , the samFor each type of crosslinking, samples differing in lecithin concentration were investigated. Lecithin concentrations were selected according to a previous study, and the samples were labeled according to these concentrations ; the reference sample without lecithin addition was marked as “R”.−1) and the negatively-charged dyes eosin B (580.09 g·mol−1) and amido black 10B (616.50 g·mol−1).The probes that were chosen to study the transport properties of the hydrogel samples differed in some of their physicochemical properties (electric charge and molecular weight) as well as in some of their visual properties, such as their absorption maximum. Specifically, the probes chosen were the positively charged rhodamine 6G . The supporting method was scanning electron microscopy (SEM), from which images of the studied hydrogels were obtained and further compared with previously published xerogel images.To characterize the transport properties of the physically crosslinked hydrogel matrix, agarose hydrogels modified by different additions of lecithin were studied. Agarose was chosen because, in the context of amphiphilic modifications, we try to approach the functionality and properties of extracellular matrix (ECM), and agarose hydrogels are frequently used in vitro models across numerous disciplines . HydrogeDeff) was the crucial transport parameter that provided us with information about the transport properties of the studied hydrogels. Specifically, it is a value that describes how each individual dye is released from the hydrogel. The procedure for obtaining Deff and the method to calculate it are described in The effective diffusion coefficient  and previous experience with this organic dye . The resWhat happens to the hydrogel after evaporation of the dispersion medium, and how does this affect the other substances in the gel? To answer this key question, the same hydrogel samples as used in the previous experiment were dried prior to characterization. In this case, however, the dye release from agarose was very quick, and it was not possible to measure the release over longer intervals as was performed for the dye release from non-dried hydrogel samples. For this reason, a fiber UV-Vis spectrometer was used, allowing the measurement of UV-Vis spectra every twenty seconds. In this setup, the absorbance was measured for approximately the first hour, which, for agarose xerogel samples, was enough time for around twenty percent of the dye to be released from the sample . As staThe images obtained from SEM show differences in hydrogel structures between samples with different lecithin concentrations  and how 2+ ions. Alginate has been extensively used as a hydrogel-synthetic ECM and thus is an ideal representative of an ionically crosslinked hydrogel for this study [As an example of an ionically crosslinked hydrogel matrix, sodium alginate crosslinked by calcium ions (2:1 wt. ratio) was studied. Bond formation resulted from the interaction of negatively charged poly(guluronic) acid units of alginate with divalent Cais study . StructuThe first studied model dye was positively charged rhodamine 6G, which was released from an ionically crosslinked alginate hydrogel. Lecithin, which acts as an internal structure modifier, was added to the hydrogel systems in different concentrations. The amount was the key factor that influenced both the hydrogel and xerogel structures and their related transport properties, which can be seen in Deff) was the highest for the reference sample and decreased with increasing lecithin concentration. The differences in Deff values for the samples with distinct additions of lecithin were negligible and thus confirmed the insignificant influence of lecithin concentration on the final value of the diffusion coefficient (in comparison with the reference sample).The amount of released dye was the highest for the reference sample, as this hydrogel did not contain any structures or artifacts that would serve as obstacles or as storage moieties for the dye, and since the alginate hydrogel complex should act as a neutral system , the dyeXerogel samples for ionically crosslinked alginate yielded very similar results to those for physically crosslinked agarose samples. In the case of lecithin additions, the concentration of dye was undetectable, probably because of the blockage of positively charged dye release caused by lecithin throughout the whole duration of the experiment. The time of development of the dye release a was almNegatively charged dyes were also studied. As is obvious from The results for ionically crosslinked xerogels with negatively charged eosin B followed those for physically crosslinked xerogels with the same dye, showing that the reference sample had a much bigger diffusion coefficient than the other samples, their lower coefficients resulting from lecithin addition. According to this, it could be concluded that crosslinking does not have a considerable influence on the diffusion coefficients of xerogels for this type of model drug (dye). The timing of the release of this dye indicates that lecithin slowed down the release since alginate does not interact with eosin B . As mentThe release of the second negatively charged dye, amido black 10B, from ionically crosslinked alginate was significantly slowed down by lecithin additions. The associated diffusion coefficient increased with increasing concentration up to the 1 wt.% sample. During the experiment, the dye release from the samples with the two highest lecithin concentrations was either immeasurable or exhibited big deviations. This was caused by disintegration, which led to the release of both the dye and the lecithin. A similar order of magnitude for the diffusion coefficients  can be oOn the other hand, ionically crosslinked xerogels with negatively charged amido black 10B show the exact same trend as physically crosslinked agarose hydrogels. The value of the diffusion coefficient decreases with increasing lecithin concentration. The results of FTIR analysis using PCA showed that the main differences between the alginate gels without and with lecithin spectrally correspond to the characteristic bands of lecithin .Prior to SEM observation, ionically crosslinked alginate hydrogels were shock-frozen in liquid nitrogen and subsequently lyophilized, as were the agarose hydrogels. As stated before, this procedure was used to minimize the alteration of hydrogel structure; however, for these types of hydrogels, it was not effective. As can be seen from A mixture of PVA ) with chitosan crosslinked by epichlorohydrin was studied as an example of a chemically crosslinked hydrogel. As well as other types of crosslinking, the internal structure and subsequent transport properties of samples were modified by the addition of different amounts of amphiphilic lecithin. Chemically crosslinked samples were studied only in their xerogel form since the preparation method finishes with mixtures in their dried state; thus, rehydrated samples would yield the same results . The onlThe main evaluated parameters were the diffusion coefficients and the amount of released dye. Three different dyes were used to study the transport properties of chemically crosslinked PVA-chitosan samples: positively charged rhodamine 6G, negatively charged eosin B, and amido black 10B. The negatively charged dyes were held inside the gel very strongly, which was because of the small pores in the highly organized structure of the chemically crosslinked gel as well as the interaction of the positively charged chitosan with the negatively charged dye. The only dye that was measurable outside the gel was released immediately after the experiment started; neither long nor short experiments, the latter focused on the first hour of the experiment, were able to provide results that could be analyzed. Only a residual amount of both of these dyes was released from the xerogels; therefore, no diffusion coefficient values could be calculated.Only experiments with the positively charged dye, rhodamine 6G, yielded results sufficient for analyses and for obtaining information about diffusion coefficients. As can be seen in The chemically crosslinked hydrogels were used in their native (hydrated) form for the preparation of samples for morphological study. The prepared xerogels were left to swell in water and were subsequently shock-frozen in liquid nitrogen and freeze-dried. The images obtained using SEM show the layered structure of the hydrogels. The only observable difference between each sample is the smoothness of the cross-sectional view. With increasing lecithin content, the interlayer roughness increases .These results correspond with already-published data for xerogels of these chemically crosslinked hydrogels . Since tThis work focused on the description of the transport properties of differently crosslinked hydrogels and xerogels , which were altered with different concentrations of amphiphilic lecithin as a structure modifier to influence the transport properties of the gels. For characterization of the transport properties of the above-mentioned hydrogel systems, various model probes with different charges were used .The positively charged dye interacted with the lecithin modifier, which meant that for the physically and ionically crosslinked hydrogels, the dye was held within the hydrogel and released very little dye or none at all, regardless of the quantity of the modifier. When the charge of the dyes did not interact with the modifier (in the case of negatively charged dyes), the quantity of lecithin had a significant impact on the speed and amount of dye release. Which meant that the diffusion coefficient was lower for the xerogel samples with lecithin compared to the reference samples. In summary, therefore, we can state that the addition of lecithin to physically or ionically crosslinked hydrogels slows down the release of the model drug and affects the amount of drug released. Chemically crosslinked hydrogels appear to be a system that is also affected by lecithin, but not as significantly as in other types of crosslinking.This work closely follows already-published results dealing mainly with the mechanical properties of the same types of hydrogels and modifications used in this work. Together with our former work, it is possible to compare different morphological characterizations of gels in their hydrogel and xerogel forms.Differently crosslinked hydrogel matrices and the effect of lecithin on their properties were studied. Three different crosslinking mechanisms were studied: physical, ionic, and chemical. Agarose E was purchased from Condalab , calcium chloride from Lach-Ner , and sodium alginate, poly, chitosan, epichlorohydrin, and L-α-Phosphatidylcholine (lecithin) were all purchased from Sigma-Aldrich . Dyes used for diffusion experiments differed in charge: positively charged rhodamine 6G , negatively charged amido black 10B , and eosin B  were used.−1, eosin B—0.015 g·l−1, or amido black 10B—0.01 g·l−1). The sample thus prepared was mixed and dissolved at an elevated temperature. It was then left to stand for 24 h to gel prior to further study [Physically crosslinked hydrogels were prepared using agarose. For every sample, a mixture of agarose (1 wt.%) and lecithin  was weighed in a container. A liquid dispersion medium was then added to the mixture. The dispersion medium was either pure deionized water  or, if needed, a solution of appropriate dye (rhodamine 6G—0.01 g·ler study .3) at a two-to-one weight ratio. Lecithin was added to the alginate solution and stirred for 24 h. A solution of calcium chloride  was sprayed on the layer of sodium alginate with lecithin in order to create a homogeneous hydrogel [Ionically-crosslinked hydrogels were prepared by sodium alginate (2 wt.%) crosslinked by calcium ions of calcium chloride  which was constantly stirred. The water was then routinely measured in a UV/Vis spectrophotometer every ten minutes to obtain the absorbance value for the released dye. The experiment ended when all the dye was released, the amount led to a constant weight, or the sample was damaged or destroyed.2∙s−1.The most fundamental parameter that can express (and consequently be used to compare) the transport properties of hydrogels with inner structural changes resulting from the addition of a modifier, such as lecithin, is the effective diffusion coefficient, expressed as the amount of dye released from the gel per mIt must be noted that calculated diffusion coefficients are not absolute values but only relative (or effective) ones. This means that porosity and tortuosity are hidden in the values of diffusion coefficients. The determination of absolute values of diffusion coefficients requires deeper research focused specifically on the diffusion phenomena of each specific hydrogel. This was not the principal aim of this publication. Therefore, we used effective values for the diffusion coefficients as proxies for comparing the transport properties of the studied hydrogels as a function of the additive concentration and/or type of crosslinking.The time of development of the mass of released dye can be expressed as follows:nt) divided by the amount of substance at equilibrium (nrov). The effective diffusion coefficients were calculated from the slope of the linear regression of this ratio of the amount of substance as a function of the square root of time. The dimensions of the releasing medium  were considered as circles of diameter (δ). From this simple calculation, it is obvious that the release experiments must be performed until the equilibrium state is achieved (when the concentration of released dye does not change over time) [The total amount of released dye at a specific time is expressed as the ratio of the amount of substance at a given time (er time) .During the measurements as well as after evaluation of the results from UV-Vis spectrophotometry, it was found that the release of the dyes from the xerogel samples was too fast for the experimental setup. For this reason, the xerogels were measured again using optical probes and OceanView 2.0 software , which allowed us to obtain UV-Vis absorbance every 20 s for the first hour of the experiment.Morphological characterization of the structures of all prepared hydrogel samples was performed using a ZEISS EVO LS 10  scanning electron microscope. The samples in their native  form were shock-frozen in liquid nitrogen and then immediately freeze-dried. A few small specimens were taken from each studied hydrogel to maintain objective observation. Before observation, the specimens were gold-coated in a sputtering device .The surface morphologies of every sample were recorded. Observations were realized in secondary electron mode, and the accelerating voltage was set to 5 kV to avoid charging the sample.


Interestingly, this magnetic coupling does not affect the intrinsic transport parameters of graphene, such as the mobility and transport scattering rate, which remain the same with and without Ni nanoparticles, indicating that the changes in the magnetotransport properties have a purely magnetic origin.We characterize the effect of ferromagnetic nickel nanoparticles (size ∼ Graphene has been one of the most studied materials over the last 20 years, due to its linear dispersion relation close to the Fermi level which gives it unique properties. Graphene exhibits a high intrinsic mobility ,2, can sThere has been some research on the magnetic properties of graphene coupled with magnetic materials. A Rashba effect has been observed for CVD graphene grown on nickel ,26, nickof 5 meV ,32. Thisof 5 meV ,34 and Hof 5 meV  in graphIn this work, we study the effect of ferromagnetic nickel nanoparticles on the magnetoresistance of graphene ribbons under applied magnetic fields of up to brikosov , is commlow 10K) ; thus, t systems ,33,34,352) layer. Standard mask photolithography using an SUSS Micro-Tec MA6 Mask Aligner and thermal evaporation using a Kurt J. Lesker PVD 75 e-Beam/Thermal Evaporator were used to define the electrode pattern. First, the wafer was prebaked at 2 surface.The substrate used was a standard 4 inch phosphorus-doped silicon wafer with a 3 bath and sonicated for 2 gas, and loaded into the CVD system. The pressure in the tube furnace was pumped down to 2 gas and 4 was used as a carbon source with a flow of Graphene was grown on a copper foil substrate  support layer of We used the hydrolysis bubble method to transfer the graphene/nickel (Gr/Ni) film from the copper foil to the Si/SiO2 gas at We fabricated the GR and Ni-GR resistors using standard mask photolithography as described above. A protective layer of PMGI was spin-coated at a spin rate of 2 and Finally, the wafer was annealed at Atomic force microscopy (AFM) images were obtained on an Asylum MFP-3D Origin microscope in AC tapping mode with a We performed the electrical characterization of the samples in a cryogen-free Versalab system from Quantum Design with applied magnetic fields of up to B, resulting inElectrical transport in graphene is commonly described with the semi-classical Drude model; however, corrections must be included in order to explain the occurrence of a field-dependent resistivity. This correction depends on EEI and is considered within the conductivity tensor in two dimensions, and does not contribute to the Hall current . The lonequation ,48,49 images are a high precision profile measurement of the height of the sample . Conversely, AFM shows in general a poor lateral resolution. There are three main factors that determine lateral precision: the range of interactions of the electrostatic potential, the cantilever tip radius, and the in-plane step size, which depends on the scan area ,57,58,59n theory ,61,62, tIndeed, the precise values of the area coverage fraction are There are two important features to observe in As the temperature increases, WL effects tend to vanish at temperatures larger than as 200 K , in agreeraction  and weakIn order to discuss the effect of the Ni NPs at high fields, in Nickel is a ferromagnetic material with a large Curie temperature in excess of 500K ,65. Smal∼0.5−1 T ,65,66. T∼0.5−1 T . The magFrom the fitting of the MR data of the Ni-GR sample, shown in nt of Ni ,69,70,71stimated ,35. Furtstimated , in agregraphene , must beOur results show two effects that are directly associated with the magnetic coupling of the graphene with the Ni nanoparticles, i.e., the appearance of a large effective exchange field and the suppression of WL phenomena. However, the question remains open whether the intrinsic electronic properties of the underlying graphene were also modified or if the Ni–C interaction is only associated with a magnetic proximity effect. We hereby study and compare the electronic transport parameters of both the GR and Ni-GR samples. From the fittings of the MR data, we were able to obtain the curvature of the longitudinal resistivity, Equation ). This qNotably, even though the MR data of the samples are very different and the fittings were carried out using two distinct models (with and without an exchange field), the curvatures resulted to be remarkably similar and essentially independent of temperature in the range 60–140 K. The inset of In Equation  both theitten asμ=e3Equation , as a fugraphene ,74. In fgraphene . This isThe magnetoresistance of graphene has a quadratic dependence with the magnetic field due to electron–electron interactions, and a zero-field peak due to a weak localization quantum effect. In this paper, we have studied the effect of ferromagnetic nickel nanoparticles on the resistivity of graphene. We show that the Ni nanoparticles produce a large magnetic exchange coupling with the underlying graphene layer. At low applied magnetic fields, the local exchange fields of the Ni nanoparticles likely introduce magnetic scattering centers, boosting the spin–orbit interaction and thus increasing the dephasing rate of the conduction electrons. The consequence is a reduction of the WL peak by a factor 3. Furthermore, at high magnetic fields the exchange coupling with the ferromagnetic nanoparticles introduces an effective field in graphene, which adds to the applied field and produces a very large MR response. However, the curvature of the parabolic magnetoresistance, controlled by the mobility and scattering time, remains the same for both the decorated and the non-decorated samples and agrees well with the expected values for graphene. From the MR curves, we obtained a Ni–graphene magnetic exchange constant of around This work demonstrates the tunability of the magnetotransport properties of graphene via a strong coupling with ferromagnetic Ni nanoparticles. Therefore, our results show the potential of graphene-based magnetic heterostructures in the design of spintronic devices.


Corresponding rhenium analogues of [99mTc]Tc-L1 were synthesised and suggested the structures of these 99mTc-complexes would be a monovalent cation with a technetium (I) core surrounded by six ligands. [99mTc]Tc-L1 is hydrophilic, while the lipophilicities of [99mTc]Tc-L2, [99mTc]Tc-L3, [99mTc]Tc-L4 and [99mTc]Tc-L5 are close. In vitro cell experiments showed that all five novel 99mTc-complexes had higher uptake in hypoxic cells compared with aerobic cells, which indicates the complexes have good hypoxia selectivity. The biodistribution of the five 99mTc-complexes in S180 tumour-bearing mice showed that they all had certain uptake in the tumours. Among them, [99mTc]Tc-L1 had the highest tumour-to-muscle (4.68 ± 0.44) and tumour-to-blood (3.81 ± 0.46) ratios. The introduction of polyethylene glycol (PEG) chains effectively reduced the lipophilicity and decreased uptake by the liver, intestine and blood but also increased clearance from the tumours. In vivo metabolic studies showed [99mTc]Tc-L1 kept intact and remained stable in tumour, blood and urine at 2 h post-injection. The results of SPECT imaging showed that [99mTc]Tc-L1 had significant tumour uptake at 2 h post-injection, but there was still high uptake in abdominal organs such as the liver and kidney, suggesting that this complex needs to be further optimised before being used for tumour hypoxia imaging.When developing novel radiopharmaceuticals, a linker moiety between the chelator and targeting vector can have a crucial influence on adjusting the affinity of the tracer and its biodistribution in organisms. To develop novel  The oxygen content in the majority of tumour tissues is notably lower than that in other tissues due to the uncontrollable growth of tumour cells that consume more oxygen than normal tissues ,2. Addit18F]fluoromisonidazole -2-nitroimidazole) is a widely used tumour hypoxia imaging agent in clinic PET imaging FMISO. Compared with 18F, 99mTc has an ideal half-life, is easy to prepare with a 99Mo-99mTc generator and comes with a reasonable cost Tc(NS3) [99mTc]Tc-4 [99mTc]Tc-2c mTc]Tc-4  and [99mTc]Tc-2c , all sho99mTc core to produce stable [99mTc][Tc(CN-R)6]+ complexes with six ligands Tc-citrate and pertechnetate as assessed by thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC), suggesting that in vitro and in vivo studies can be carried out without further purification. The proposed structure of these complexes is monovalent cation with a 99mTc core in the centre surrounded by six identical ligands, which are similar to that of [99mTc]Tc-sestamibi (sestamibi: 2-methoxy-2-isobutyl isonitrile). The predicted chemical structures of the five complexes are shown in 99mTc]Tc-L1 were synthesised and identified by 1H-NMR and MS to verify the structures of the 99mTc-complexes, as both Tc and Re belong to the ⅦB group and have similar chemical properties. The HPLC profile of [99mTc]Tc-L1 co-injected with Re-L1 is shown in 99mTc]Tc-L1 (tR = 9.61 min) was close to the UV pattern of Re-L1 (tR = 8.94 min), indicating that [99mTc]Tc-L1 had the proposed structure.By reaction of pertechnetate with the reducing agent, stannous chloride, provided in the kit, the 99mTc-labelled complexes were kept in saline at r.t. or in mouse serum at 37 °C for 4 h, the RCPs of them were all higher than 90%, suggesting that these complexes do not decompose within 4 h and have good stability in vitro.The RCP of these complexes can be used to evaluate the stability of the complexes. As shown in 99mTc]Tc-L1 is hydrophilic, while the lipophilicities of [99mTc]Tc-L2, [99mTc]Tc-L3, [99mTc]Tc-L4 and [99mTc]Tc-L5 are close. Compared with [99mTc]Tc-L2, which contains a carbon chain (n = 7), [99mTc]Tc-L1 containing a polyethylene glycol  chain has a lower Log D value, indicating that [99mTc]Tc-L1 is more hydrophilic and the addition of PEG chains can effectively improve the hydrophilicity. By comparing [99mTc]Tc-L3, [99mTc]Tc-L4 and [99mTc]Tc-L5, it was found that the lipophilicities of the three 99mTc-complexes with phenyl ring were close, indicating that there was no positive correlation between the increase in the number of carbon atoms on both sides of the phenyl ring in these substituents and the lipophilicity of these 99mTc-complexes.The Log D values of the five tracers are shown in p < 0.05, in The in vitro cellular uptake of five tracers was tested on S180 cells under both aerobic and hypoxic conditions. 99mTc-complexes are shown in 99mTc]Tc-L2, containing a carbon chain (n = 7), had the highest tumour uptake value (1.22 ± 0.22%ID/g) and good retention in tumours for 2 h (1.19 ± 0.24%ID/g). However, the high uptake of [99mTc]Tc-L2 in the liver (33.31 ± 3.75%ID/g) and kidney (24.70 ± 2.77%ID/g) may increase the difficulty in the detection of abdominal tumour. By comparing the 99mTc-complexes containing phenyl rings , it was found that [99mTc]Tc-L4 had the highest tumour uptake and good retention, with an uptake of 0.97 ± 0.13%ID/g at 2 h, and the highest tumour/muscle ratio, which reached 4.15 ± 0.54 at 2 h post-injection. Among these three complexes containing phenyl rings as linkers, uptake in the liver and spleen increased with the reduced numbers of carbon atoms. For [99mTc]Tc-L5, the uptake in the liver at 0.5 h was 45.08 ± 8.90% ID/g and the uptake in the spleen was 36.16 ± 3.80%ID/g. In fact, these 99mTc-complexes containing phenyl rings are not ideal tumour hypoxia imaging agents because of their slow clearance in vivo and high uptake in nontarget organs, including the liver, kidneys and spleen. In comparison, [99mTc]Tc-L1 containing tetra-oxyethylene PEG chains had the highest tumour-to-muscle (4.68 ± 0.44) and tumour-to-blood ratios (3.81 ± 0.46), and the ratios of the tumour to other nontarget tissues were significantly reduced, indicating that [99mTc]Tc-L1 was more favourable than the other four 99mTc-complexes for further research.The results of biodistribution studies of the 99mTc]Tc-L1 was selected to conduct SPECT/CT imaging studies because of its high target-to-nontarget ratios. SPECT/CT images are shown in 99mTc]Tc-L1 at 0.5 h, 1 h and 2 h post-injection are shown in [99mTc]Tc-L1, S180 tumour-bearing mice were used for metabolic studies in tumour, blood, and urine at 2 h post-injection. As shown in To investigate the in vivo metabolism of Tc-L1 were synthesised and identified by 1H-NMR and MS to confirm that the structure of the 99mTc-complexes is a monovalent cation with a 99mTc core in the centre surrounded by six identical ligands. The five 99mTc-complexes all exhibited excellent in vitro stability. The Log D values of the five 99mTc-complexes showed that [99mTc]Tc-L1 had the highest hydrophilicity. In vitro cell experiments showed that all five novel 99mTc-complexes had higher uptake in hypoxic cells compared with aerobic cells, which indicates the complexes have favourable hypoxia selectivity. Female Kunming mice bearing S180 tumours, which were used as a model of solid tumour with hypoxia [99mTc]Tc-L1 (1.11 ± 0.13% ID/g) containing a PEG chain was slightly lower than that of [99mTc]Tc-L2. However, the tumour retention of [99mTc]Tc-L1 at 2 h post-injection was only 40% of that at 0.5 h post-injection, and the fast clearance from the tumour might relate to its hydrophilicity. The introduction of the PEG chain effectively reduced the lipophilicity and decreased the uptake in the liver, intestine and blood, but it also accelerated the clearance from the tumours [99mTc]Tc-L1 had the highest tumour/muscle (4.68 ± 0.44) and tumour/blood (3.81 ± 0.46) ratios, indicating the potential of this compound for further research. To investigate clearance in vivo, we studied the biodistribution at 6 h post-injection of [99mTc]Tc-L1. With faster clearance in kidneys and intestine and some retention of the tracer in tumour, [99mTc]Tc-L1 shows a preferable target-to-nontarget ratio. Study of the in vivo metabolism of [99mTc]Tc-L1 was carried out by analysis of the radioactivity in different tissues using HPLC. The results showed that the tracer remained mostly intact and thus was stable in vivo. Only small percentages of metabolites were found in blood, urine and tumour. As linkers can significantly alter the biodistribution potency of compounds, the purpose of this study was to design, synthesise and evaluate novel  hypoxia ,33, were tumours ,35. Inte99mTc-2c, a 99mTc-labelled 2-nitroimidazole derivative containing a carbon chain (n = 4), was reported to have a good tumour-to-muscle ratio (5.05) Tc-L1 containing a PEG chain as the linker showed a lower retention in blood and exhibited for this reason a higher tumour-to-blood ratio at 2 h post-injection (T/B for [99mTc]Tc-L1: 3.81 vs. T/B for 99mTc-2c: 1.32). Compared with the cyclopentadienyl tricarbonyl technetium-99m 2-nitroimidazole derivative [99mTc]Tc-4, [99mTc]Tc-L1 showed higher tumour-to-muscle ratio (T/M for [99mTc]Tc-L1:4.68 vs. T/M for [99mTc]Tc-4: 3.47) and tumour-to-blood ratio (T/B for [99mTc]Tc-L1:3.81 vs. T/B for [99mTc]Tc-4: 0.59) Tc-L1 had higher tumour-to-blood and tumour-to-muscle ratios than the other four tracers, it was chosen as a prospective tracer for further SPECT/CT imaging studies. The SPECT/CT imaging results of [99mTc]Tc-L1 exhibited observable tumour uptake, which was consistent with the results of biodistribution studies in mice. However, the relatively high uptake of [99mTc]Tc-L1 in the liver and kidneys is a shortcoming of the complex, and the results of abdominal tumour imaging may be affected. In future studies, we will focus on improving the tumour uptake and retention of hypoxic tumour molecular probes and reducing their uptake in the liver and kidneys by changing the linkers and the strategy of coordination.As [99mTc]NaTcO4 was eluted from a 99Mo-99mTc generator which was manufactured by Curium Netherlands B.V. . ESI-MS spectra were acquired by a Triple TOFTM 5600 spectrometer . The IR spectra were obtained by an IR-Affinity-1 spectrometer . 1H-NMR and 13C-NMR spectra were acquired on a JNM-ECS spectrophotometer . Radioactivity was assessed using an HRS-1000 technetium analyser  and a Wizard 2480 γ-counter . HPLC analysis was carried out with an analytical column  and a SHIMADZU system  equipped with an SPD-20A UV detector (λ = 254 nm) and a Bioscan flow count 3200 NaI/PMT γ-radiation scintillation detector . SPECT/CT imaging was recorded on a Triumph SPECT/CT scanner . Female Kunming mice (18–22 g) were obtained from Beijing Vital River Laboratory Animal Technology, Beijing, China.All chemicals in the studies were obtained from commercial sources and used without purification. +: found 414.1975, calcd 414.1983.Ligand L2 (8-isocyano-N-(2-(2-nitro-1H-imidazol-1-yl)ethyl)octanamide): white solid, yield 87%. 1H NMR  δ 7.49 , 7.11 , 4.40 , 3.44 , 1.93 , 1.52 , 1.36 , 1.31–1.24 , 1.23–1.12 ; 13C NMR  δ 173.09, 155.87, 145.21, 129.02, 128.08, 49.65, 41.65, 38.56, 35.65, 28.91, 28.35, 26.13, 25.45; IR (KBr)/cm−1: 3309.09, 3078.52, 2930.00, 2856.70, 2148.79, 1653.07, 1539.26, 1489.11, 1363.73, 1273.07, 1165.05, 1082.11; HR-MS (ESI) for C14H22N5O3 [M + H]+: found 308.1721, calcd 308.1717.Ligand L3 (2-(4-(isocyanomethyl)phenyl)-N-(2-(2-nitro-1H-imidazol-1-yl)ethyl) acetamide): yellow oily liquid, yield 56%. 1H NMR  δ 7.32 , 7.25 , 7.18 , 7.03 , 4.39 , 3.45 , 3.31 , 2.46 ; 13C NMR  δ 170.97, 156.87, 145.15, 136.44, 132.06, 130.05, 128.86, 128.09, 127.36, 49.58, 45.23, 42.28, 40.19; IR (KBr)/cm−1: 3315.78, 3105.53, 2146.86, 1637.63, 1535.40, 1477.54, 1357.94, 1276.93, 1153.48, 1089.83; HR-MS (ESI) for C15H16N5O3 [M + H]+: found 314.1245, calcd 314.1247.Ligand L4 (4-(isocyanomethyl)-N-(2-(2-nitro-1H-imidazol-1-yl)ethyl)benzamide): white solid, yield 75%. 1H NMR  δ 7.79–7.66 , 7.49–7.40 , 7.34 , 7.06 , 4.81 , 4.72–4.59 , 3.82 ; 13C NMR  δ 168.59, 156.03, 137.04, 133.77, 127.67, 127.55, 127.11, 126.65, 49.24, 44.40, 39.15; IR (KBr)/cm−1: 3364.00, 2945.43, 2835.48, 2150.72, 1651.14, 1548.91, 1479.49, 1356.02, 1286.58, 1253.78, 1161.20, 1028.10; HR-MS (ESI) for C14H14N5O3 [M + H]+: found 300.1090, calcd 300.1091.Ligand L5 (4-isocyano-N-(2-(2-nitro-1H-imidazol-1-yl)ethyl)benzamide): white solid, yield 63%. 1H NMR  δ 7.03–6.90 , 6.75–6.65 , 6.54 , 6.25 , 3.89–3.82 , 3.01 ; 13C NMR  δ 165.69, 155.72, 135.49, 129.23, 128.18, 126.98, 49.55, 39.01; IR (KBr)/cm−1: 3358.21, 3275.27, 2922.28, 2852.84, 2125.65, 1653.07, 1539.26, 1489.11, 1363.73, 1282.72, 1165.13, 1097.54; HR-MS (ESI) for C13H12N5O3 [M + H]+: found 286.0939, calcd 286.0934.Ligand 99mTc-complexes, the ligand was first dissolved in ethanol for obtaining a solution with a concentration of 5 mg/mL. Then, 0.1 mL of the solution (containing 0.5 mg of the ligand) was added to the reagents of a kit . Freshly prepared [99mTc]NaTcO4  was added to the kit. It was shaken and heated at 100 °C for 20 min. The radiochemical purity was measured by TLC and HPLC. TLC was conducted using a polyamide strip  with methanol as developing solvent. The target 99mTc-complexes moved to the top (Rf = 0.8–1.0), while [99mTc]NaTcO4 and 99mTcO2·nH2O stayed at the origin (Rf = 0–0.1). For HPLC, phase A was water, phase B was acetonitrile with a flow rate of 1 mL/min. The gradient elution profile was as follows: 0–2 min 10% B, 2–5 min 10–90% B, 5–20 min 90% B, 20–24 min 90–10% B, 24–25 min 10% B. Retention times of the 99mTc-complexes: 9.54 min ([99mTc]Tc-L1); 9.52 min ([99mTc]Tc-L2); 9.50 min ([99mTc]Tc-L3); 9.72 min ([99mTc]Tc-L4); and 9.51 min ([99mTc]Tc-L5).To prepare the 99mTc]Tc-L1 structure, the stable complex Re-L1 was synthesised. Re-L1 was prepared as follows: 100 mg of ligand L1 was dissolved in 0.5 mL normal saline, then 1.5 mL sodium citrate buffer solution  and 150 μL stannous chloride solution (50 mg/mL) were added and 5 mg potassium perrhenate was added. The reaction mixture was stirred at 100 °C for 3 h. The reaction solution was separated and purified by HPLC to obtain the rhenium complex. The identification method of HPLC was the same as the method in 1H NMR and MS. 1H NMR  δ 7.38 , 7.11 , 4.54 , 3.62–3.50 , 2.34 ; MS: [(C17H27N5O7)6ReNa2H]3+: m/z calcd 904.69, found 904.69.To validate the proposed Tc-L1 .SPECT/CT imaging studies were performed using a Triumph SPECT/CT scanner, reconstructed with HiSPECT software  and processed with Vivoquant 2.5 software . The mice used for the SPECT/CT imaging studies were anaesthetised and maintained under anaesthesia during the imaging procedure. The SPECT/CT imaging was performed with female Kunming mice bearing S180 tumours at 2 h post-injection of [99mTc]Tc-L1  was injected into Kunming mice bearing S180 tumours via a tail vein. After 2 h, the mice were sacrificed, then urine, blood and tumour were collected. The urine was passed through a 0.22 µm microporous membrane and analysed by HPLC. The blood was centrifuged at 10,000× g for 3 min, then 100 μL of the supernatant was mixed with 200 μL of acetonitrile. After centrifugation, the protein was precipitated and removed, and the supernatant was collected and passed through a 0.22 µm microporous membrane, then it was concentrated and analysed by HPLC. The tumours were homogenised by a TH-02 tissue-tearor  with 200 μL saline, then 400 μL of acetonitrile was added into the centrifuge tube and the mixture was centrifuged to precipitate the protein, and the supernatant was collected after centrifugation. Then, the supernatant was passed through a 0.22 µm microporous membrane, concentrated by rotary evaporator and analysed by HPLC. The method of HPLC was the same as the method in Tc-L1 had the highest hydrophilicity and the highest tumour-to-muscle and tumour-to-blood ratios. And it also stayed intact and remained stable in tumour, blood and urine at 2 h post-injection. The SPECT/CT imaging results showed that [99mTc]Tc-L1 was obviously taken up in the tumour site, but there was still the problem of high uptake in the liver and abdomen. Further exploration of structural modifications of the ligand and the linker is necessary.In this study, five isonitrile hypoxia ligands with different linkers were designed, synthesised and radiolabelled with


Here, we analyse the behavioural configuration that people adopt stereotypically when ashamed – slumped posture, downward head tilt, gaze avoidance, inhibition of speech – in light of shame's hypothesised function. This behavioural configuration may have differentially favoured its own replication by (a) hampering the transfer of information (e.g. diminishing audiences’ tendency to attend to or encode identifying information – shame camouflage) and/or (b) evoking less severe devaluative responses from audiences (shame display). The shame display hypothesis has received considerable attention and empirical support, whereas the shame camouflage hypothesis has to our knowledge not been advanced or tested. We elaborate on this hypothesis and suggest directions for future research on the shame pose.The shame system appears to be natural selection's solution to the adaptive problem of information-triggered reputational damage. Over evolutionary time, this problem would have led to a coordinated set of adaptations – the shame system – designed to minimise the spread of negative information about the self and the likelihood and costs of being socially devalued by others. This  Social media summary: The shame display functions to appease others. Landers and Sznycer propose an additional function: to render us invisible.Shame occurs in every known culture Brown, ; its comAdaptations are packages of design features retained over evolutionary time because they reliably solved tasks tributary to reproduction in ancestral environments. An adaptationist approach to understanding shame thus begins from theories of the adaptive problems our ancestors faced over deep evolutionary time , audiences react by attaching less weight to that person's welfare  devaluation reliably elicits shame  traits  will be less prone to shame all else equal, because their superior position can be leveraged to (a) impose or threaten to impose more costs on others to incentivise less devaluation or prevent it altogether . Furthermore, shame within each country tracked the magnitude of devaluation reported in the other two countries, suggesting universality in both the structure and content of shame .This reasoning led to empirical tests of the hypotheses that the shame system is designed to (a) accurately forecast the magnitude of audience devaluation on an act-by-act basis, and (b) generate an internal signal of anticipatory shame in proportion to that forecast. Across three different cultures , participants were presented with a set of socially devalued acts or traits  and were asked to rate in a between-subjects design, for each act or trait, either the intensity of shame they would feel if the act or trait were true of them or how negatively they would view another individual if the act or trait were true of that other individual . In all three countries, the intensity of anticipatory shame closely tracked the magnitude of devaluation expressed by local audiences a–c. FurtThe hypothesis that shame's adaptive function is to mitigate the threat of devaluation can explain many published findings on shame. For instance, being devalued causes people pain  effects for gaze avoiders: indeed, when an individual commits a moral infraction of which observers are aware, individuals who subsequently gaze-avoid are judged more favourably than those who gaze-seek. And audiences’ responses toward individuals known to have transgressed are more forgiving when the transgressor produces the gaze-avoiding shame display than when the transgressor fails to produce gaze-avoidance  on audiences has been interpreted as appeasement , suggest ways the shame system could realise this function, and offer questions that future research could address.While a sizeable literature deals with the appeasing function of the shame display, no research we are aware of has elaborated on the possibility that the shame ‘display’ might serve another function: evasion from identification . We use quotation marks around ‘display’ here because according to this hypothesis, the characteristic constellation of shame actions functions not to elicit a forgiving response from audiences, but to evade recognition and evaluation by the audience. That is, in some contexts the shame ‘display’ may function as camouflage rather than as a signal The view of the shame ‘display’ as a means to achieve invisibility can be found in Darwin's Ring of Gyges, Tolkien's The One Ring, Rowling's Cloak of Invisibility, Wells’ Invisible Man and Marvel Comics’ Invisible Woman, to the Daoist texts on goddess Jiutian Xuannü and Persian mythology's Khrafstra.Incidentally, we note that the hunger for invisibility – to act free of the threat of others’ devaluation and aggression, to gain information about others unmolested by reputational concerns, to snoop around without being regarded as a snoop  may seem odd or hyperbolic, but shame can render people invisible The shame system can produce invisibility in various ways: leading one to forgo actions that may result in being devalued, interrupting one's actions when detection by others is imminent, hiding, fleeing, equivocating, deceiving, justifying one's actions, and concocting other ruses for concealing oneself in plain sight. How might the static shame ‘display’, or camouflage, confer invisibility? We suggest two possible ways.One way in which the shame camouflage could confer invisibility is by disrupting observers’ ability to identify the ashamed individual. Data exist which are consistent with this possibility. For example, observers are quicker to find faces in an array  gaze are better remembered by observers  to render shamed individuals invisible to observers – that is, to interfere with observers’ ability to identify or recall ashamed individuals and thus to devalue them. In what follows, we offer a series of questions to help guide future research on the invisibility hypothesis.How quickly do observers identify targets who adopt the characteristic static shame pose  vs. a neincrease observer identification of and/or memory for individuals exhibiting the shame pose relative to other expressions. Indeed, in reactive mode, the shame system may produce a shame display designed to call attention to one's submission, coupled with characteristic verbal and nonverbal actions drawing attention to one's acceptance of lower status (e.g. saying ‘I'm sorry’). In prospective mode, however, before observers know that a transgression has occurred, the shame pose may reduce observer identification and/or memory compared with other kinds of emotional expressions. To corroborate the invisibility hypothesis, research must demonstrate reduced observer ability to identify and/or recall individuals exhibiting the shame pose relative to other emotional expressions given at least one level of observer knowledge.The hypothesised invisibility effect of the shame pose on both memory and identification is likely to interact with observer knowledge: how would learning the identity of the culprit who committed the shame-inducing act alter observer identification of and/or memory for individuals who exhibit the shame pose relative to other emotional expressions? Learning that a shameful event has occurred but lacking knowledge of the culprit, or lacking knowledge of any shameful event occurring in first place, may also uniquely affect observer identification and/or memory for individuals exhibiting the shame pose relative to other emotional expressions. Precisely because the shame display may also function as a signal of appeasement, learning that a shameful event has occurred, but lacking knowledge of the culprit, may in the kinds of high-coresidence, tightly knit ecologies in which our ancestors evolved. Therefore, an important test of the invisibility hypothesis is whether known community members  attain invisibility dividends when they adopt the shame pose. The attainment of invisibility by known community members seems less probable, because observers possess a wealth of background knowledge that can aid in identifying familiar transgressors , and more is at stake for observers when identifying actual wrongdoers in their community compared to when identifying the fictional wrongdoing of strangers. Yet this high evidentiary bar needs to be met for the invisibility hypothesis to show viability.According to the invisibility hypothesis, invisibility is conferred by adaptive design. This assumes that net benefits from invisibility would have accrued over deep time on average to those who adopted the shame pose in the right place at the right time Answers to these and related questions will corroborate or falsify the invisibility hypothesis.An adaptationist approach to shame paints a unique picture of this emotion: rather than a maladaptive source of hostility and psychopathology, shame is a messenger, one that defends the individual by transmitting devaluation-relevant information to an array of countermeasures against devaluation. The information threat theory can explain many known facts about shame and generate novel predictions. Some of these have received broad support across cultures (Sznycer et al.,


Pseudomonas aeruginosa; however, comparative data are lacking. The purpose of this study was to compare rates of tx-emergent resistance among patients (pts) with bacteremia or pneumonia at a single center.CZA and CT are front-line agents for treatment (tx) of MDR Adult pts treated for >48 hours with CZA or CT were included. Pts with cystic fibrosis or colonization were excluded. Isolates were tested by broth microdilution (BMD) in triplicate and underwent whole-genome sequencing.Table 1). CT-treated pts were less likely to receive prolonged infusions and monotherapy, but more likely to have empyema/endovascular infections. Baseline median (range) MICs were 4 (2–8mg/L) and 2 (0.25–8mg/L) for CZA and CT, respectively. Within 90-days from tx onset, rates of resistance defined as either ≥4-fold MIC increase or MIC >8mg/L were higher among CZA-treated pts (Table 1). Corresponding median MICs of resistant isolates were 16 (16–128mg/L) and 64 (32–512mg/L), respectively. Across 107 baseline isolates, 59 different sequence types (ST) were represented. The most common P. aeruginosa cephalosporinase (PDC) variants were PDC-3, PDC-5, and PDC-8, present in 10%, 21%, and 10% of isolates, respectively. Resistance evolved across varying STs. A subgroup of contemporary pts was well-balanced and supported overall findings (Table 2). From this subgroup, rates of tx-emergent resistance among pts receiving CZA or CT monotherapy were 50% (6/12) and 6% (1/17), respectively (P=0.01). Rates of CT-resistance were lower among pts who received prolonged  versus standard  infusions. Paired baseline and resistant isolates showing ≥4-fold MIC increase were compared (Table 3). CZA resistance was associated with sequence changes in ampD and/or efflux genes in 5 pts; 2 additional pairs showed mutations in ftsI (PBP3). CT resistance was associated with new ampC mutations in 83% of pairs.113 pts were included. Demographics, severity of illness, and durations of tx were similar for pts tx’d with CZA or CT : Advisor/Consultant|LabSimply: Advisor/Consultant|Merck: Advisor/Consultant|Shionogi: Advisor/Consultant|Shionogi: Honoraria Ryan K. Shields, PharmD, MS, Allergan: Advisor/Consultant|Cidara: Advisor/Consultant|Entasis: Advisor/Consultant|GSK: Advisor/Consultant|Melinta: Advisor/Consultant|Melinta: Grant/Research Support|Menarini: Advisor/Consultant|Merck: Advisor/Consultant|Merck: Grant/Research Support|Pfizer: Advisor/Consultant|Roche: Grant/Research Support|Shionogi: Advisor/Consultant|Shionogi: Grant/Research Support|Utility: Advisor/Consultant|Venatorx: Advisor/Consultant|Venatorx: Grant/Research Support


Estimating insertion depth, cochlear duct length (CDL), and other inner ear parameters is vital to optimizing cochlear implantation outcomes. Most current formulas use only the basal turn dimensions for CDL prediction. In this study, we investigated the importance of the second turn parameters in estimating CDL. Two experienced neuro-otologists blindly used segmentation software to measure (in mm) cochlear parameters, including basal turn diameter (A), basal turn width (B), second-turn diameter (A2), second-turn width (B2), CDL, first-turn length, and second-turn length (STL). These readings were taken from 33 computed tomography (CT) images of temporal bones from anatomically normal ears. We constructed regression models using A, B, A2, and B2 values fitted to CDL, two-turn length, and five-fold cross-validation to ensure model validity. CDL, A value, and STL were longer in males than in females. The mean B2/A2 ratio was 0.91 ± 0.06. Adding A2 and B2 values improved CDL prediction accuracy to 86.11%. Therefore, we propose a new formula for more accurate CDL estimation using A, B, A2, and B2 values. In conclusion, the findings of this study revealed a notable improvement in the prediction of two-turn length (2TL), and CDL by clinically appreciable margins upon adding A2 and B2 values to the prediction formulas. Since the implementation of universal neonatal screening programs in most developed countries, patients with SNHL have been able to benefit from early intervention4. The gold standard for most cases of SNHL is the cochlear implant (CI). CIs are a safe and cost-effective treatment option for patients with severe-to-profound SNHL who derive insufficient benefit from hearing aids. CI use is an effective means of enabling patients to develop  hearing and communication abilities6.Sensorineural hearing loss (SNHL) is the most frequent form of permanent hearing impairment and has an estimated prevalence of 0.13% among neonates in developed countries. In adolescents, this prevalence increases to 0.35% due to accumulated late-onset genetic and acquired cases8. Advancements in CI are moving toward offering (electrode) arrays of varying lengths to account for individual variations in CDL, rather than using “one size” arrays designed for cochleae of average size. Such individualization can be used to maximize treatment outcomes and improve users’ quality of life. The intracochlear electrode array is placed adjacent to peripheral dendrites of the afferent auditory nerve and spiral ganglion cell bodies (SGCBs) in the scala tympani. Recently, Dhanasingh et al.10 demonstrated that SGCBs are distributed in the basal turn and extend into the second turn as well, with segment I  containing only 11.3% of SGCBs and segments II and III  containing 63%. However, segment IV (angular depth of up to 680°) contains 25.8% of SGCBs. There is a need to be precise in determining the balance between safe atraumatic insertion and optimum insertion depth of the array to maximize the coverage of the cochlea while preserving the hearing structures, thereby maximizing CI outcomes.For optimal outcomes, CIs must be tailored to each recipient's requirements. Cochlear shape, size, and length vary among individuals; this geometrical variation can reach up to 30–40%11. Currently, the most clinically viable methods to estimate the CDL and angular insertion depth are dependent only on cochlear basal turn parameters, which can be measured by different methods described in the literature15. The basal turn of the cochlea, while readily identifiable, does not provide a comprehensive representation of the cochlear structure due to its highly variable nature among individuals16. This is particularly critical for CIs, where an accurate assessment of CDL can influence array choice and placement, ultimately impacting hearing outcomes17. Furthermore, the middle turn contains a large fraction of the SGCBs, which play a pivotal role in CI stimulation and overall auditory function18. As a result, incorporating the second-turn parameters in CDL estimation could both lead to more accurate measurement and facilitate tailored approaches to CI, improving post-operative hearing performance. Thus far, no method has been developed to measure the second-turn parameters and incorporate them in the CDL. Moreover, the middle and apical turns are not always predicted by basal turn parameters7.A deeper understanding and knowledge of cochlear architecture and morphology, and concise measurements of cochlear parameters will aid in better pre-operative estimation of cochlear duct length (CDL) and angular insertion depth. This mitigates the effects of incomplete array insertion, migration, folding, or kinking. Furthermore, it will yield a more precise allocation of frequency bands to electrode contacts, thereby producing maximum benefits and performance11. Although, sub-optimal cochlear implantation may not solely be due to inaccuracies in CDL prediction. Factors such as training and experience of the surgeon as well as the characteristics of the array itself play important roles in the success of the insertion process.Although many CI centers use CDL preoperatively to select the array length that best fits the recipient’s cochlear architecture, there are still some CI cases with challenging and incomplete insertion or over-insertion. Thus, there is a need for a new method that more accurately estimate the CDLTherefore, this study aimed to investigate the importance of the second-turn parameters in estimating CDL. The secondary objective was to test the accuracy of our new proposed formulas in predicting more accurate CDL and 2TL.This retrospective study was conducted at a tertiary CI center. Pre-operative CT scans of temporal bones were retrieved from the medical image database of King Abdullah Ear Specialist Center, King Saud University. All CT scans that showed a good compatibility with the 3D reconstruction software and resulted in a clear and high-resolution 3D model have been included to the study regardless age, gender, side. Patients were excluded if they had poor CT image resolution, in case of any inner ear anomalies, or if they had middle or inner ear pathologies. This study was conducted in accordance with the Declaration of Helsinki and approved by the local institutional review board of King Saud University (No. E-22-6574). The same local committee waived the need for informed consent due to the retrospective nature of the study. Routine clinical data, including age, sex, ear side, and array type, were collected.https://www.slicer.org/) following the previously reported steps11. The software’s four-window layout allowed us to manipulate and examine the image in axial, coronal, sagittal, and 3D segmentation views, the model was freely manipulated to allow for optimal visualization. The slice plane was adjusted parallel to and over the 3D segmentation, providing a consistent perspective for measurements. To standardize this approach, the 3D model was projected into a Cartesian coordinate system, with the X, Y, and Z axes set perpendicular to the cochlear aperture and the modiolus.All CT images in the present study were obtained using a 512-slice multidetector-row CT scanner . The following scanning parameters were used: axial plane, 0.625 mm slice thickness, 230 mAs, 140 kV, and rotation time 1 s with 0.3 mm reconstruction in the axial and coronal views. DICOM images of patients’ temporal bones were imported and segmented using the 3D Slicer multi-platform software  represented the largest linear distance of the basal turn from the center of the round window to the farthest point on the opposite of the lateral wall, passing through the modiolus. The B value (width) represented the width of the basal turn of the cochlea, measured as a straight-line running perpendicular to the A value line at the modiolus. We defined A2 as the diameter of the second turn, which was from the start of the second turn at 360° to the furthest point of the second turn through the modiolus. B2, which represented the width of the second turn, was measured as a line perpendicular to A2 that passed through the modiolus. CDL was measured manually by tracing a curve line from the center of the round window to the cochlear lateral wall up to the most apical bony point of the apex. The basal turn length (BTL) was measured manually as a curve line from 0° to 360° of the cochleae. Then, STL which was the second or mid-turn was measured as a curve line from 360° to 720° of the cochleae. Finally, the two-turn length (2TL) was both the basal and mid-turns of the cochlea from 0° to 720° of the cochleae.The A and B values were measured in the cochlear view projection, as previously described by Escude et al.8. The B2/A2 ratio is introduced and used in this study to represent the ratio of the width to the diameter of the second turn to determine the cochlear shape.The B/A ratio represented the ratio of the width to the diameter of the basal turn to determine the corresponding round/elliptical shapeAll statistical analyses were performed using Python version 3.9. Briefly, we determined the ICC between the two readers to validate their measurements. Parametric data are presented as mean ± SD, whereas nonparametric data are presented as median . For inferential statistics, we performed bivariate comparisons of the variables grouped by sex. We used the two-sided independent sample t-test for parametric data and the Mann–Whitney U test for nonparametric data. Furthermore, we used Pearson's correlation to estimate correlations between the B/A ratio to the B2/A2 ratio to assess cochlear dimensional consistency across patients.2, and test accuracy from the cross-validation for each model.To validate the importance of measuring A2 and B2 values for predicting CDL, and 2TL, we fitted the A, B, A2, and B2 values to predict them using multiple linear regression models. We compared these models to those fitted to the A and B values alone and to those fitted to A2 and B2 values alone. To further validate the results of these regression models, and to account for the small sample size, we performed a five-fold cross-validation and calculated the regression coefficients, mean RThe study was conducted in accordance with the Declaration of Helsinki and approved by the local institutional review board of King Saud University (No. E-22-6574).The aforementioned ethics committee waived the need for informed consent due to the retrospective nature of the study.CT scans of 17 patients (33 ears) aged 1.6 (1.1–2.9) years with good resolution were eligible for the measurement of cochlear parameters. All implantations were with a MED-EL electrode array : 15 ears received a FLEX28, 11 received a FORM24, 3 received a FLEX24, 3 received a STANDARD, and 1 received a MEDIUM. Table We calculated the intraclass correlation coefficient (ICC) across the two measurements done by the two readers. A, A2, and B2 values showed ICC values consistently above 0.6, which were always significant . However, the B value showed moderate ICC values, which were also significant . When assessing normality, only age, A2 value, and STL variables were not normally distributed ; therefore, nonparametric tests were used to test those variables going forward.The B/A ratio ranged from 0.70 to 0.87 , whereas the B2/A2 ratio ranged from 0.76 to 0.99 . The B/A ratio was not significantly correlated with the B2/A2 ratio . In contrast, the A value exhibited a significant moderate positive correlation with B  and A2 . B showed a similar correlation with B2 , and A2 showed a similar correlation with B2 . Figure The simple linear regression model showed that A2 has a significantly higher correlation  than B2  with STL  was lower than that mentioned by the same team 25. These results suggest that cochlear size in our population is proportionally different from that of other populations. Additionally, Erixon et al. made their measurements on temporal bone corrosion casts, which provide a high level of detail and accuracy in capturing cochlear dimensions. In contrast, our study relied on 3D segmentation of clinical CT images. While CT imaging provides valuable insights into cochlear anatomy, it may introduce certain limitations compared to direct measurements on corrosion casts.In our sample, BTL mean and range  represent 61% of the total cochlear length, whereas STL mean and range  account for 32% of the cochlear length, which was lower than the mean lengths for BTL (22.6 mm) and STL (12.4 mm) reported by Erixon et al.8, we found that the second turn tends to be more frequently round, regardless of the shape of the basal turn in the same cochlea, due to the high B2/A2 ratio and significantly small variation (SD = 0.06). To explore these relationships, we might require a larger sample size with more variations in B2/A2 ratios to study the shapes of the second turn in depth.In our study, although the B/A ratio was not significantly correlated with the B2/A2 ratio, A and B showed moderate correlation with A2 and B2, respectively. This indicates significant morphological variability that is constrained by the linear lengths and widths in the studied cohort. Notably, compared with the basal turn, which can have an oval or round shape according to the B/A range as demonstrated by Khurayzi et al.Regression analyses showed that A2 or B2 can be used to predict STL, with A2 being the better predictor: for each 1 mm increase in A2 and B2 values, the STL increased significantly by approximately 1.23 and 0.90 mm, respectively. Further, adding the A2 and B2 values to our regression models improved the prediction accuracy for 2TL and CDL by approximately 3%. However, fitting regression models to a small sample size produces inconsistent results. Additionally, running the model several times can produce markedly different results, which means that A2 and B2 can be as important for CDL prediction, which might be revealed with a larger sample size.We performed five-fold cross-validation across our models to minimize the inconsistency caused by the small sample size. Our analysis showed that the A and B values were consistently significant when predicting STL and CDL; the higher the A and B values, the higher the patients' CDL and STL. Surprisingly, these results did not extend to the A2 and B2 values, despite capturing features that should be associated with STL.10. Therefore, it is crucial to optimize array insertion depth to improve patient outcomes. Our results contribute to the body of evidence that can help surgeons increase the accuracy of CDL prediction.Various CI centers worldwide agree that most SGBCs are in the third and fourth cochlear segments14 adapted the geometric modeling formula by Yoo et al.26, and used the A value and insertion angular depth to approximate the lateral wall length of the cochlea. They showed that the larger the A value, the higher the insertion angle depth and the longer the lateral wall length14. More recently, Schurzig et al.15 developed an elliptic-circular approximation algorithm to predict the insertion length and angles in each individual achieving higher accuracy estimation. The available equation-based models often include a predetermined representation of data based on the helical shape of the cochlea. Our study uses a validated multiple linear regression modeling approach; although the data representation is predetermined, it has more room to learn from the data provided, thereby producing predictions that fit the confounding population differences.Escudé et al.27, which influences the accuracy of A2 and B2 measurements, and by extension, the computation of 2TL, and CDL. Secondly, the technique used for measuring A2 and B2 can also affect the calculation. The precision of these measurements may be impacted by the resolution of the imaging technique, the angle at which measurements are taken, and potential observer variability in identifying anatomical landmarks28. Thirdly, our regression models assume a linear relationship between A2 and B2 values and cochlear lengths. This may not fully capture the complexity of the true relationship, particularly given the three-dimensional spiral structure of the cochlea29. Lastly, the demographic characteristics of the sample population, such as ethnic background, may affect the generalizability of the model and its prediction accuracy in different groups. Future research could focus on quantifying and minimizing the impact of these influencing factors, potentially through the use of automated measurement techniques, advanced imaging technologies, and stratified analysis according to demographic characteristics. Additionally, the use of more complex mathematical models may be warranted to reflect the intricate relationships more accurately between various parameters and cochlear lengths.The process of calculating CDL and incorporating the second-turn parameters, A2 and B2, in the model is intricate, with a number of factors exerting influence. Firstly, the inherently variable nature of cochlear anatomy among individuals is one key consideration. Notably, the size, shape, and orientation of cochlear turns can vary greatlyThis study was limited by the relatively small sample size which might affect its generalization. Our measurements were taken from the lateral wall of the cochlea, which, while providing a practical and consistent landmark, does not necessarily correspond directly to the distance that the array would travel within the scala tympani during cochlear implantation. A radial correction factor to convert the cochlear length estimates from the lateral wall to the estimated array location within the scala tympani was not incorporated into our study.We acknowledge that the potential for observer variability cannot be eliminated and may influence the computation of various lengths. Future work with bigger sample size should consider conducting an error analysis to quantify variability in slice angles and how such variation might affect measurements of A2 and B2, and ultimately the calculated lengths.We found that the second turn of the cochlea tends to be round more frequently regardless of the basal turn shape. The findings of this study revealed a notable improvement in the estimation of 2TL, and CDL by clinically appreciable margins upon adding A2 and B2 values to the prediction formulas. Thus, our model and formulas may help clinicians choose the proper electrode-array length before the CI surgery. This improvement comes at little to no cost and may be detrimental to some patient outcomes.


Hypnosis can be an effective treatment for many conditions, and there have been attempts to develop instrumental approaches to continuously monitor hypnotic state level (“depth”). However, there is no method that addresses the individual variability of electrophysiological hypnotic correlates. We explore the possibility of using an EEG-based passive brain-computer interface (pBCI) for real-time, individualised estimation of the hypnosis deepening process.The wakefulness and deep hypnosis intervals were manually defined and labelled in 27 electroencephalographic (EEG) recordings obtained from eight outpatients after hypnosis sessions. Spectral analysis showed that EEG correlates of deep hypnosis were relatively stable in each patient throughout the treatment but varied between patients. Data from each first session was used to train classification models to continuously assess deep hypnosis probability in subsequent sessions. Models trained using four frequency bands  showed accuracy mostly exceeding 85% in a 10-fold cross-validation. Real-time classification accuracy was also acceptable, so at least one of the four bands yielded results exceeding 74% in any session. The best results averaged across all sessions were obtained using 1.5–14 and 4–15 Hz, with an accuracy of 82%. The revealed issues are also discussed.The online version contains supplementary material available at 10.1186/s13104-023-06553-2. Hypnosis can be an effective treatment for numerous disorders –3. The hProbably, a hypnotic state gradually evolves during a session and tends to fluctuate . The posPassive brain-computer interfaces (pBCI) are used to assess mental states such as fatigue, concentration, etc. , 59. We Nine outpatients  underwent up to seven hypnosis sessions. Inclusion criteria: age 18–65 years; consent for participation. Patients had previously reported experiencing deep hypnosis, described as a lack of self-awareness, external awareness, and memories for the deepest period of a session. Thus, probably all included participants could be classified as “dissociators” –62, “amnThe BCI system included equipment for synchronised EEG and video recording and also software: WinEEG 2.130.101 , EEGLab In each patient, after installing the electrodes and equipment for EEG and video recording, a baseline EEG was recorded for 3–5 min with eyes closed. Hypnosis was then induced and deepened by the counting method. After therapeutic suggestions, a patient was awakened. Feedback was then collected.We used the passive type of BCI , 59, 70 The physical signs recommended to verify sufficient depth , 24, 26:The patients’ post-session feedback on the hypnotherapist’s counting range during which they felt most deeply hypnotised (see “Supplements A” for thiThe file with two sets of labels was then used to train a classifier to continuously recognise (“predict”) these two opposite states in subsequent sessions with interdependent probability. Assuming that deepening is a continuous transition from wakefulness to the deepest hypnotic state, we hypothesise that the continual real-time measurement of the probability of a deep hypnosis during a session could tentatively, to some extent, operate as a quantitative reflection of the deepening process.The collected data were processed in four ways.Using WinEEG to compare the averaged power spectra of the identified deep hypnosis and the waking periods (5–10 min each) in each EEG, we obtained an overview of each patient’s assumed deep hypnosis patterns . We also assessed their putative stability throughout the treatment, qualitatively comparing them over different patient sessions. This analysis was conducted in parallel with all the others.During supervised learning, we trained prediction models, and those derived from the first sessions were then used in the following sessions for real-time classification. The Common Spatial Pattern (CSP) –76 methoUsing models derived from the first sessions for real-time state predictions in subsequent sessions yielded a Probability Value parameter, varying between 0 and 1, displayed as a moving curve, which informed the hypnotherapist of the probability of a deep hypnotic state occurring. We called this curve the Predictive curve. “Supplements A” describEach second and subsequent session was labelled by a specialist not privy to their predictive outcomes, and a percentage of correctly predicted states for different epochs was calculated to additionally test the accuracy . The aboPatient T reported no periods of deep hypnosis in the first session and was excluded from the analysis. Due to the artefacts, the recording from session #5 of Patient E was also excluded. Thus, the total number of EEGs from the remaining eight participants was 27.Figure As seen, the electrophysiological changes tend to be generally similar in different sessions of a particular patient, which suggests that a classifier would correctly recognise the target patterns in the following sessions. Correlates common to the patients are seen, e.g., a slow-wave activity increase, which is consistent with the literature , 45–57. “Supplements B” contain all the maps for all 27 sessions for all patients and the details on electrophysiological changes revealed.Table As an example of such results, Fig. “Supplements C” contain the Predictive curves for each patient. In general, post-session patients’ reports revealed an approximate match between the time when a curve was consistently above 0.7  and a period of unawareness. The high-amplitude wave-like motion was associated with alternating awareness, fractional memories, etc. Thus, this could potentially confirm instrumentally the literature reports of hypnotic depth variations during a session . “SuppleTable The accuracy averaged across all these sessions was highest when using bands 1.5–14 and 4–15 Hz. This is also in line with studies that reported the most changes in alpha and theta activity , 54, 55.Figure 2(B) shows the Native curve of Patient A’s seventh session. The configurations of curves (A) and (B) largely coincide, giving us additional confirmation that the model can reflect the real picture relatively accurately. “Supplements C” contain the pairs of the Predictive and Native curves for each patient for the second and following sessions.This study extends the idea of a “hypnometer”  but focuThe designed system is a trial version only and requires further substantial improvements, using both the results and issues we revealed. However, we suppose it could initially demonstrate that pBCI applies to hypnosis.Small sample size, the heterogeneous number of sessions, and the artefacts in some recordings.Involvement only of those patients who described their deepest experiences as a lack of awareness and self-awareness. Although these phenomena are common for hypnotic experiences , 65, 66,We did not measure the participants’ hypnotisability. Trait effects are considered to have a different basis –7, 9, anA single rater was used to label each given recording.Using the measurement of the continuously changing deep hypnosis probability as a tentative reflection of the deepening process is a hypothetical idea in the early stages of testing. The qualitative analysis conducted could partly underpin the information from a Predictive curve, but it is not comprehensive. Further studies, utilising the classification of intermediate levels of hypnosis, are needed.The qualitative analysis of estimated hypnotic patterns is approximate, and further studies can incorporate statistical assessment to strengthen these findings as well as quantify differences in accuracies across subjects and bands and identify similarities between the Native and Predictive curves.Our system is a prototype only, and the signal processing techniques used are not comprehensive, being the initial option. Further research is necessary to test other feature extraction approaches and classification methods , 82.Below is the link to the electronic supplementary material.Supplementary Material 1(“Supplements A”)Supplementary Material 2 (“Supplements B”)Supplementary Material 3 (“Supplements C”)Supplementary Material 4 (“Supplements D”)


Researchers are increasingly seeking to use routinely collected data to support clinical trials. This approach has the potential to transform the way clinical trials are conducted in the future. The availability of routinely collected data for research, whether healthcare or administrative, has increased, and infrastructure funding has enabled much of this. However, challenges remain at all stages of a trial life cycle. This study, COMORANT-UK, aimed to systematically identify, with key stakeholders across the UK, the ongoing challenges related to trials that seek to use routinely collected data.This three-step Delphi method consisted of two rounds of anonymous web-based surveys and a virtual consensus meeting. Stakeholders included trialists, data infrastructures, funders of trials, regulators, data providers and the public. Stakeholders identified research questions or challenges that they considered were of particular importance and then selected their top 10 in the second survey. The ranked questions were taken forward to the consensus meeting for discussion with representatives invited from the stakeholder groups.In the first survey, 66 respondents yielded over 260 questions or challenges. These were thematically grouped and merged into a list of 40 unique questions. Eighty-eight stakeholders then ranked their top ten from the 40 questions in the second survey. The most common 14 questions were brought to the virtual consensus meeting in which stakeholders agreed a top list of seven questions. We report these seven questions which are within the following domains: trial design, Patient and Public Involvement, trial set-up, trial open and trial data. These questions address both evidence gaps  and implementation gaps (requiring training and/or service re-organisation).This prioritised list of seven questions should inform the direction of future research in this area and should direct efforts to ensure that the benefits in major infrastructure for routinely collected data are achieved and translated. Without this and future work to address these questions, the potential societal benefits of using routinely collected data to help answer important clinical questions will not be realised.The online version contains supplementary material available at 10.1186/s13063-023-07251-x. Routinely collected data (RCD) is increasingly being used to support health research, including clinical trials, in the UK and worldwide. RCD include data collected for purposes other than research, including electronic health records (EHR)—the digital version of patient health information  and registry data—a collection of information about individuals, usually focused on a specific diagnosis or condition. The use of RCD in randomised controlled trials has the potential to bring essential benefits to patients participating in clinical trials, healthcare staff supporting clinical trials and members of the public and researchers globally who may be impacted by the trial findings .Accessing data directly from the RCD sources is expected to reduce the burden on patients and care staff otherwise involved in collecting data directly from individuals (bespoke data collection). RCD has the potential to provide data that are more scientifically valid (i.e. long-term data follow-up and high levels of completeness), enhance generalisability (i.e. high retention and more inclusive and diverse sample) and reduce the overall cost to the public of conducting large-scale research studies. Despite growing recognition of such potential benefits of using RCD for trials research, there remain key methodological challenges for researchers with identifying, accessing and using these datasets in practice and several areas of uncertainty –6.Beyond publications of expert opinion and limited examples reporting the (intended) use of RCD for trials, there is very little published evidence to guide researchers in this field. As a result, there remain broad uncertainties over the better approaches to ensure the highest standard of scientific rigour for use of RCD in trials. Anecdotally, these uncertainties include data and linkage quality, access and regulatory compliance, ethical considerations, public involvement and privacy.Identifying the top uncertainties in this area will direct efforts to ensure that the benefits in major infrastructure for RCD (in particular EHRs) are achieved and translate into benefits for methodology and efficiency in future trials, as well as for patients and their clinical teams. Without this, the potential societal benefits of using RCD to help answer important clinical questions will not be realised. The scope for this work will need to be wide as different countries’ systems and infrastructures vary considerably.This study aimed to systematically identify the current methodological research questions and uncertainties related to the use of RCD in trials, from the perspective of relevant stakeholders in the UK.We conducted a three-step Delphi method , 8 consiParticipants were eligible for the study if they were a UK stakeholder as defined in Table We used non-probability purposive sampling methods to sample survey respondents . The tarRespondents were asked to supply methodological research questions they felt were of particular importance in relation to RCD for trials. Methodological research questions were clarified as issues that can be answered with research methods and a visual map of the study life cycle was available to prompt respondents to think about all aspects of a research project (without giving specific examples which could have biassed responses) . The presentation order was randomly changed for each respondent so that all responses had an equal chance of being selected. A free-text box asked if there were any questions missing for respondents to add in as relevant. The survey was live for 3 weeks . After the closure of the survey, the questions were then ranked based on their frequency of appearing in the respondent’s top 10. The rankings were reviewed for the whole sample and then separated by stakeholder groups to identify any striking differences between stakeholder groups that may have skewed the whole sample ranking. The highest-ranking questions were taken forward to the consensus meeting, in addition to the responses provided in the free text (i.e. respondents suggested missing questions).We held a virtual consensus meeting in May 2022. Stakeholders from each group  connections. All responses were transferred to Cardiff University secure servers and analysed within Nvivo 12 (survey 1) and Microsoft Excel (survey 2). Mentimeter® votes were collected anonymously, and the results were saved on the same university servers.n = 10).Survey 1 was completed by 66 respondents. The most frequently reported stakeholder group self-identified by respondents was “trialist” (75%). The proportion of respondents from each stakeholder group, geographic spread and type of routinely collected data is shown in Table The second survey presented the 40 questions for respondents to select their top 10. There were 88 responses with the majority being trialists (65%), 30% reported completing survey 1, 23% were unsure and 40% had not completed survey 1. Table All 40 questions were included in the top 10 of at least 5 respondents, with the highest-ranked question being included by 50 respondents  and implementation gaps (requiring training and/or service re-organisation). These top questions have been published as a freely available infographic.n = 92 and n = 77 [n = 790) and priority II on retention (n = 456) [The numbers of responses for the two surveys were 66 and 88. These are comparable to other recent Delphi studies with similar stakeholder groups or scope , for example, CONSORT-ROUTINE which received d n = 77 . Howevern = 456) , these rn = 456) , and theAlthough all stakeholder groups were represented in both surveys, our sample’s composition was dominated by trialists despite our broad engagement strategy. Trialists will, however, have the most experience in all stages of the trial lifecycle  and therefore inherently have more to input in such a focussed Delphi study. Two-thirds of respondents of the second survey either did not contribute to the first survey or were unsure if they had done so and were provided with the opportunity to add in any questions at the end of the survey they considered were missing from the list of 40 questions presented. Only three questions emerged which were subsequently re-categorised during the consensus meeting within one of the 40 questions. This is an indication that additional input from stakeholders may not have yielded substantially different results to those reported here. It is acknowledged that the method of collection prevented a calculation of a response rate between surveys 1 and 2. In addition, we had representation from stakeholders across all four countries in the UK, at all stages of academic career, with expertise in using different types of data sources.methodological  was provided at the start of this survey. However, a considerable proportion of responses related to operational or implementation challenges require new or clearer guidance rather than new methodology work. Through discussion with the wider study team, it was agreed that all questions should be taken forward in the second survey as some that appeared operational may indeed be addressed via methodological research. This, combined with the additional subjectivity of such categorisation, meant that in practice, it was felt inappropriate to separate them into two groups.The first survey focussed on remaining questions that were methodological in nature. A definition of This study focused on UK stakeholders as a starting point, and the findings will only reflect the methodological questions from a UK standpoint. It is important to recognise that some of the key questions are likely to reflect structures and data that vary by country but that some of the short and long-list of challenges are likely to have some relevance to other countries, which may represent scope for future work.These prioritised questions require input from all stakeholders included in this study and beyond. Investment in infrastructure supporting RCD needs to be accompanied by a commitment to understanding the best methods and best practices for the use of RCD in trials. Data providers and regulators in particular will play a large role in addressing the two questions related to streamlining approvals across organisations and expediting timelines for data acquisition. These rely on working together to adapt their organisational processes and/or streamlining across organisations. Funders will continue to be crucial in supporting methodological projects aiming to address many of these questions. Recognition that we need to better understand these questions for the greatest benefit to patients, care teams and the public is gaining traction. Health Data Research (HDR) UK has recently funded a number of time-limited projects that will contribute to answering some of these seven questions and will be well placed to take many of these remaining questions forward over the coming years along with other key stakeholders such as the MRC-NIHR Trials Methodology Research Partnership. Members of the public and public contributors on trials will have particular significance in contributing to communication and building trust in trials that use RCD. Finally, trialists will  push these questions forward and provide input to the data provider and regulator consultations to ensure trials are included in strategic decisions and organisational prioritisation. The MCR-NIHR Trials Methodology Research Partnership plays a crucial role in this.This prioritised list of seven questions will inform the direction of future research in this area, and direct efforts should be made to ensure that the benefits in major infrastructure for routinely collected data are achieved and translated. It is important to consider the findings in light of the limitations presented and that the research priorities are dynamic over time , so any research priority exercise will need to be refreshed periodically. Without this and future work to address these questions, the potential societal benefits of using routinely collected data to help answer important clinical questions will not be realised.Additional file 1. Survey 1: Guidance provided and the format the question was asked.Additional file 2. Full list of 40 questions included in survey 2.Additional file 3. Final ranking of all 40 questions.Additional file 4. Infographic. Also, available on study page: https://www.cardiff.ac.uk/centre-for-trials-research/research/studies-and-trials/view/comorant-uk.