The United States of America is currently in an opioid epidemic. Heroin remains the most lethal opioid option with its death rate increasing by over 500% in the last decade. The rewarding and reinforcing effects of heroin are thought to be mediated by its ability to increase dopamine concentration in the nucleus accumbens shell. By activating Gi/o‐coupled μ‐opioid receptors, opioids are thought to indirectly excite midbrain dopamine neurons by removing an inhibitory GABAergic tone. The partial μ‐opioid receptor agonist buprenorphine is a substitution‐based therapy for heroin dependence that is thought to produce a steady‐state level of μ‐opioid receptor activation. But it remains unclear how buprenorphine alters dopamine release relative to heroin and how buprenorphine alters the dopamine‐releasing effects of heroin. Because buprenorphine is a partial agonist at the μ‐opioid receptor and heroin is a full agonist, we predicted that buprenorphine would function as a weak dopamine releaser relative to heroin, while functioning as a competitive antagonist if administered in advance of heroin.We performed fast‐scan cyclic voltammetry in awake and behaving rats to measure how heroin, buprenorphine HCl, and their combination affect transient dopamine release events in the nucleus accumbens shell. We also performed a complimentary pharmacokinetic analysis comparing opioid plasma levels at time points correlated to our neurochemical findings.Both buprenorphine and heroin produced changes in the frequency of transient dopamine release events, although the effect of buprenorphine was weak and only observed at a low dose. In comparison with vehicle, the frequency of dopamine release events maximally increased by ~25% following buprenorphine treatment and by ~60% following heroin treatment. Distinct neuropharmacological effects were observed in the high‐dose range. The frequency of dopamine release events increased linearly with heroin dose but biphasically with buprenorphine dose. We also found that buprenorphine pretreatment occluded the dopamine‐releasing effects of heroin, but plasma levels of buprenorphine had returned to baseline at this time point.These findings support the notion that low‐dose buprenorphine is a weak dopamine releaser relative to heroin and that buprenorphine pretreatment can block the dopamine‐releasing effects of heroin. The finding that high‐dose buprenorphine fails to increase dopamine release might explain its relatively low abuse potential among opioid‐dependent populations. Because high‐dose buprenorphine decreased dopamine release before occluding heroin‐evoked dopamine release, and buprenorphine was no longer detected in plasma, we conclude that the mechanisms through which buprenorphine blocks heroin‐evoked dopamine release involve multifaceted pharmacokinetic and pharmacodynamic interactions. Buprenorphine functions as a weak dopamine releaser relative to heroin but its pretreatment occludes the dopamine‐releasing effects of heroin. These data may help explain why buprenorphine poses minimal risk for abuse yet functions as an effective substitution therapy for opioid dependence. The partial μ‐opioid receptor agonist buprenorphine is thought to act as a substitution therapy in opioid dependence; the μ‐opioid receptor antagonist naloxone is primarily added to Suboxone® in order to prevent intravenous (IV) buprenorphine abuse.The United States of America is currently in an opioid epidemic that is producing over 100 deaths per day. Fentanyl and heroin are the most lethal opioid options, with their respective death rates increasing by 520% and 533% between 2009 and 2016.Heroin produces its rewarding and reinforcing effects by increasing dopamine concentration in the mesolimbic dopamine pathway,By investigating the effects of opioids on dopamine transients, we sought to build upon the existing microdialysis, electrochemistry, and electrophysiology literature. Previous publications show that heroin increases dopamine concentration in the NAc shellTo test these predictions, we used fast‐scan cyclic voltammetry (FSCV) to measure drug‐induced changes in dopamine release in the awake and freely moving rat. While monitoring transient release events in the NAc shell, we treated rats with ascending intravenous (IV) doses of either: heroin alone or buprenorphine followed by heroin Figure . To furt22.1Catheterized male Long‐Evans rats, supplied by Charles River, were singly housed under a 12:12‐h light‐dark cycle with a 10 AM to 10 PM active period (dark phase). All experiments were conducted in the active phase. Rats (275‐325 g at the time of surgery) were placed under isoflurane anesthesia  for surgery conducted in a stereotaxic apparatus. A guide cannula that mates with a micromanipulator was implanted to be aimed at the NAc shell . The shell region of the NAc was targeted because the initial effects of drugs on dopamine concentration are most pronounced in this region.2.22 and electrolytic lesions were performed to confirm electrode placement. Brains were extracted and frozen in −25°C 2‐methylbutane and then stored at −80°C until they were coronally sectioned at 50 μm using a cryostat. Slices were dehydrated with baths of increasing ethanol concentration, preserved with histoclear, and mounted for observation of lesion placement.Voltammetric recordings were conducted by lowering a glass‐encased carbon fiber microelectrode using a micromanipulator that fits inside the implanted guide cannula. An initial waveform  was applied which allowed for the detection of dopamine from cyclic voltammograms taken every 100 ms. To increase electrode sensitivity, the waveform was first applied at 60 Hz for ~30 minutes but reduced to 10 Hz before experimentation. To extract the dopamine component, principle component regression was applied to the raw voltammetric data.2.3As previously described,p1, p2, p3) with the largest R2 value were assigned for each individual 60 seconds dopamine concentration file. The degree of the polynomial was determined by finding the lowest Akaike information criterion (AIC) score with the following equation:The coefficients (A second fitted line was generated to establish a relative zero (“baseline”) using the following equation:To reduce type 1 error, only peaks above 1.5 standard deviation from the “baseline” that were greater than 0.5 seconds apart were analyzed. If multiple peaks occurred within the 0.5 seconds period, the highest concentration event was reported.2.4Heroin and buprenorphine were prepared in sterile saline and briefly sonicated. In all experiments, opioids were administered intravenously using a cumulative‐dosing approach. For the heroin‐only condition, we administered vehicle  and then 0.27, 0.55, 1.12, and 2.24 heroin (mg/kg IV). Cumulative doses were administered every 10 minutes as illustrated in Figure In the buprenorphine‐heroin condition, we administered vehicle  and then ascending doses of buprenorphine: 0.011, 0.044, 0.177, 0.708 Figure . Continu2.5In the present study, we took advantage of a unique clinical investigation in which human participants were given ascending doses of intravenous buprenorphine and heroin—in addition to several other opioids, and then asked to subjectively rate the resulting experience/affect (eg “I feel good”).The Km used for human = 37; the Km used for rat = 6.2.2.6g for 12 minutes to separate components. Separated plasma was collected and sent to the Pharmacology core at Colorado State University for combined liquid chromatography/mass spectroscopy (LC‐MS), which consists of a Shimadzu HPLC machine with a Waters Sunfire C18 5 µm 4.6 × 150 mm elution column (Part No. 186002557) coupled to a 3200 Q‐TRAP triple quadrupole mass spectrometer .To determine circulating concentrations of buprenorphine, heroin, and its metabolites, we performed a pharmacokinetic analysis. Long‐Evans rats were either treated with cumulative heroin doses only or pretreated with cumulative buprenorphine doses followed by cumulative heroin doses. The cumulative‐dosing scheme for heroin and buprenorphine  was identical to the voltammetry experiment dosing scheme previously described. 0.5 mL of whole blood was taken from animals via femoral vein catheter and placed in KDTA‐coated blood collection tubes at regular time intervals postdrug infusion. For the heroin‐only group Figure , blood w2.6.1g and transferred to an autosampler with inserts.A 0.1 mg/mL stock solution of buprenorphine, morphine, M3G, and 6‐acetylmorphine (6‐AM) in 50/50 ACN/Milli‐Q was prepared. Standard curves were created using the following concentrations  ng/mL buprenorphine, morphine, M3G, and 6‐AM in 50/50 ACN/Milli‐Q. A 100 ng/mL solution of fentanyl in 50/50 ACN/Milli‐Q was prepared as an internal standard. Next, 5 µL of each standard was added to fresh 1.5‐mL microcentrifuge tubes, along with 5 µL of 500 ng/mL fentanyl solution. 9 Quality control samples  ng/mL were prepared in the same manner. 50 µL of blank Rat plasma and 50 µL of acetonitrile with 0.1% formic acid were added to each tube and vortexed for 5 minutes. The tubes were then centrifuged for 15 minutes at 20 000 2.6.2g, and transferred to an autosampler vials with inserts.Five microlitre of 50/50 ACN/Milli‐Q, 5 µL of 100 ng/mL fentanyl standard, 50 µL of unknown experimental rat plasma sample, and 50 µL of ACN with 0.1% formic acid were added to fresh 1.5‐mL microcentrifuge tubes. The tubes were vortexed for 5 minutes, centrifuged for 15 minutes at 20 000 2.7Because parametric statistical tests such as ANOVA assume that experimental samples have equal variance with data that are normally distributed, we first performed tests of normality and equal variance. If data were determined to normally distribute and show equal variance, we assessed for significant differences in the data using parametric statistics . In contrast, if data were determined to not fit these criteria, we proceeded to assess for significant differences in the data using nonparametric statistics. Final statistical analyses were performed using SigmaPlot 11.33.1To assess the effects of heroin, buprenorphine, and their interaction on transient dopamine release events, we treated rats with cumulative ascending IV doses of either heroin‐alone (n = 6) or buprenorphine followed by heroin (n = 7) while concurrently performing FSCV in the freely moving rat. Doses were adapted from a clinical study that asked opioid‐dependent humans to compare the subjective effects of ascending doses of buprenorphine  and heroin . Our calculated rat animal equivalent doses (AEDs) were  for buprenorphine and  for heroin.3.2P < .05. All doses were tested cumulatively, with 10 minutes elapsing between each IV treatment. A one‐way repeated‐measures ANOVA revealed that heroin F4,29 = 14.12, P < .05  changed the frequency of dopamine release events. Tukey post hoc analysis revealed that the 2.2 mg/kg heroin dose significantly increased the frequency of dopamine release events vs vehicle and the 0.27 mg/kg heroin (P < .001). In addition, the 1.1, 0.55, and 0.27 mg/kg heroin doses all increased the frequency of dopamine release events vs. vehicle  to increase the frequency of dopamine release events . Tukey post hoc analysis further revealed that the 0.55, 1.1, and 2.2 mg/kg cumulative IV doses of heroin significantly increased 6‐AM versus vehicle. We next treated naïve rats (n = 6) with cumulative doses of buprenorphine followed by cumulative doses of heroin . Tukey post hoc analysis further revealed that the 2.2 mg/kg cumulative IV dose of heroin significantly increased 6‐AM versus vehicle (P < .05). We also measured buprenorphine plasma levels in the buprenorphine + heroin group after each series of cumulative opioid treatments and found that buprenorphine plasma levels returned to baseline by the end of the final cumulative heroin treatment . Tukey post hoc analysis further revealed that the 0.7 mg/kg cumulative dose of buprenorphine significantly increased buprenorphine plasma content (P < .05) but buprenorphine plasma content was not distinguishable from vehicle 10 minutes following the final cumulative heroin treatment—a time point in which buprenorphine pretreatment prevented heroin‐evoked dopamine release.To measure just how much buprenorphine and 6‐acetylmorphine  is in plasma at different time points in our cumulative‐dosing scheme, we performed an additional pharmacokinetic (PK) assay. We first treated naïve rats with the same cumulative doses of heroin used in our FSCV study Figure  and withn Figure . An unpan Figure  and 6‐AMn Figure  demonstr4In the present study, we used FSCV to measure how heroin, buprenorphine, and their combination affect transient dopamine release events in the NAc shell of freely moving rats. While these unique opioids both produced dose‐dependent changes in the frequency of dopamine release events, their effect on the amplitude of each event was less resolved. Analysis of the frequency data revealed that low‐dose buprenorphine functions as a weak dopamine releaser relative to heroin. When compared to vehicle, the frequency of dopamine release events maximally increased by ~25% following buprenorphine treatment and by ~60% following heroin treatment. However, distinct neurochemical effects emerged in the high‐dose range. While heroin lawfully increased the frequency of dopamine release events in a linear dose‐dependent manner, buprenorphine dosage produced a bell‐shaped effect on release. The frequency of buprenorphine‐evoked dopamine release events increased across the 0.01‐0.04 mg/kg range before declining to a level of insignificance across the 0.18‐0.7 mg/kg range. This latter observation confirms a previous microdialysis report demonstrating that buprenorphine alters accumbal dopamine concentration in a biphasic, dose‐dependent manner.The opioid doses used in the present study were adapted from a double‐blind, placebo controlled inpatient trial. We selected these doses so that we could then compare our neurochemical findings to subjective responses that are impossible to measure in animal models. Similar to the present study, Comer and colleaguesThree classical opioid receptors  exist.The complimentary PK analysis offers additional insight into the complex pharmacokinetic and pharmacodynamic mechanisms through which buprenorphine alters opioid‐evoked dopamine release. While the PK data confirm that our cumulative‐dosing scheme lawfully increased plasma levels of 6‐AM, we were surprised to find that buprenorphine was no longer detectable in plasma at a time point in which its pretreatment occluded heroin‐evoked dopamine release. This observation is supported by the findings of Ohtani et alThe μ‐opioid receptor‐mediated increase in accumbal dopamine concentration is theorized to arise from disinhibition of dopamine neurons in the VTA.®/Suboxone® would produce distinct effects on dopamine release vs IV buprenorphine. Opioid bioavailability would also be significantly lower following SL vs IV buprenorphine. Future studies should be conducted to compare the interaction of buprenorphine and heroin on opioid‐evoked dopamine release in rats with a history of contingent drug self‐administration because the neurochemistry of acute noncontingent administration of drugs to animals is quite different than what occurs in models of reinforcement such as self‐administration. Thus, while it may be worth considering that Subutex®/Suboxone® prescription guidelines suggest that physicians begin treatment with 8 mg/70 kg sublingual (SL) buprenorphine—a dose that is comparable to the highest buprenorphine dose used in the present study (0.7 mg/kg AED in rat), it is likely that IV buprenorphine is more potent and effective at increasing dopamine release vs. the same dose of SL buprenorphine.While we attempted to make our dose assessments relevant for the clinical literature, caution should be used during the extrapolation of our dose‐dependent effects. Aside from being rats, our sample is different from that used by Comer et alNotwithstanding these limitations, our data provide new insight into how clinically relevant doses of heroin, buprenorphine, and their combination influence dopamine release. While heroin dose‐dependently increased the frequency of dopamine release events, buprenorphine only increased dopamine release in the low‐dose range. In addition, we found that buprenorphine pretreatment attenuates the dopamine‐releasing effects of heroin. Because high‐dose buprenorphine stopped increasing dopamine release and buprenorphine was no longer present in plasma, it is likely that its ability to blunt heroin‐evoked dopamine release is not exclusively a result of buprenorphine competing with 6‐AM at the μ‐opioid receptor.We have no conflicts to disclose.EBO designed the experiment and obtained funding to conduct it. DPI, RPL, TJE, HL‐B, and EBO performed the experiments and statistical analysis; DPI, LRH, and EBO wrote the manuscript. LRH and EBO finalized the manuscript. All authors read and approved the final manuscript.We need not require IRB review.We did not require informed consent.We did not register a clinical trial for this study.The University of Colorado Denver Institutional Animal Care and Use Committee approved all animal experiments and procedures in advance.


This work investigates the effect of replacing normal aggregate either partially or totally with expanded perlite aggregate. This material allows for better thermal insulation properties, thus decreasing the energy usage within the life cycle of the concrete structure. Expanded perlite aggregate was used in concrete by 20%, 40%, 60%, 80%, and 100% in replacement of the natural aggregate. Material characterization tests of compressive strength, flexural strength, and thermal conductivity were carried out for six concrete mixtures. In addition, microstructure analysis was performed with the aid of a micro-computed tomography system to investigate the effects and relation of microstructure quantities on material properties. The proposed concrete mixture, which has 100% of expanded perlite aggregate, has a unit weight of 1703 kg/m In missions ,6. Previmissions . Therefomissions .It has been demonstrated that energy efficiency measures such as thermal insulation retrofitting are more cost-effective compared with the harvesting of renewable sources such as solar energy and wind energy ,10. Conv−1·K−1, which is fairly high and thus exhibits poor insulation properties [Because concrete is the most commonly used building material, researchers have explored methods for enhancing the properties of concrete materials ,12,13. Toperties ,15. The operties ,17.−1·K−1 but significantly decreased the compressive strength to 20 MPa. In an attempt to rectify such tradeoffs, significant research has been dedicated to investigating the use of perlite and expanded perlite (EP) material in concrete.In a previous study , the eff−1·K−1 although at the expense of a much lower compressive strength of 3.71 MPa. In addition, the effects of EP on the mechanical and thermal properties of lightweight concrete were tested in [The popularity of perlite and EP is attributed mainly to its desired thermal conductivity properties and light weight. Perlite is a hydrated amorphous- volcanic silicate glass whose volume rapidly expands between 5 and 20 times of its original volume when heated about 900–1200 °C, forming EP ,22,23. Fested in . In thatested in  evaluateested in ,28,29,30Although a significant amount of research has focused on the development of lightweight, thermally insulated concrete, the methods proposed in the literature have major limitations. First, most of the adjusted concrete mixtures have low mechanical strength, which renders them unacceptable for structural use. Second, the adjusted concrete mixtures with somewhat improved properties are suitable only for use in building envelopes such as blocks and plaster. The literature lacks the development of a concrete mixture that exhibits high mechanical properties and low thermal conductivity. Therefore, in the present study, we evaluate the influence of EP on the thermal and mechanical properties of high-performance concrete. The motivation behind this study is the development of a novel, lightweight concrete mixture that exhibits reduced thermal conductivity while retaining strong mechanical properties that enable its use in structural elements such as load bearing walls, columns, and roof slabs.Motivated by the above, the work presented herein undertakes an experimental evaluation of an adjusted concrete mixture that incorporates various fraction of EP within the mix is conducted in this study. The reference concrete mixture is composed of ordinary Portland cement, microsilica, fly ash, river sand, and polypropylene microfibers. The replacement of sand with EP is applied in increments of 0%, 20%, 40%, 60%, 80%, and 100% of the volume of the sand. The resulting mixtures are tested to evaluate their mechanical properties such as compressive and flexural strengths, thermal properties such as thermal conductivity, and microstructural characteristics via X-ray computed tomography (CT).For the experiments undertaken in this work, ordinary Portland cement, microsilica, fly ash, river sand with a maximum particle size of 2 mm, EP with a maximum particle size of 4 mm, and polypropylene microfibers with a length:diameter ratio of 12:0.18 mm were employed. The polypropylene microfibers were used to reduce the shrinkage and deformation in the plastic state. Polycarboxylic ether superplasticizer, provided by BASF, was used to maintain concrete workability with a low water/cement ratio (W/C).Particle size grading of the sand, cement, fly ash, and microsilica was conducted using a laser diffraction analyzer . However, due to the large size of the EP aggregate (>2 mm), sieve analysis was implemented as per the ASTM C136/C136M-19 standard. 3, respectively. Six concrete mixtures were prepared with the same W/C ratio of 0.28 to test the effects of replacing normal aggregate with EP aggregate in increments of 0%, 20%, 40%, 60%, 80%, and 100% of the volume of the sand. The cementitious binder remained the same for all mixtures, containing 70%, 20%, and 10% of ordinary Portland cement, fly ash and microsilica, respectively. The manufacturer’s recommended dosages of superplasticizer and microfiber were used, at 1 L/kg of cementitious material and 1.2 kg/m3 and decreased approximately in a linear rate to 1700 kg/m3 for mixture 6 that has 100% EP.3 cube, resulting in a ~25.4 mm ø cylindrical sample with a length of ~50mm, as illustrated in Fifteen samples were taken from each of the six mixtures for testing purposes. Twelve of these cubic samples, with dimensions of 50 × 50 × 50 mm, were used to test the compressive strength at different time intervals of 1, 7, 28, and 56 days. To evaluate the flexural strength, three beam specimens with dimensions of 40 × 40 × 160 mm were prepared and tested on day 28. Fragments that developed during the flexural strength testing were used for thermal conductivity analysis. One of the three remaining samples from the original 15 for mixtures 1, 4, and 6 was used for microstructural analysis using X-ray CT imaging. The samples were cored from the center of each 50 mmThe mechanical properties were tested according to ASTM standards. The compressive strength of three samples representing each mixture was tested on days 1, 7, 28, and 56 with a loading rate of 1200 N/s according to ASTM C109/C109M−16a. The arithmetic average was reported for each test. The flexural strength of each mixture was evaluated by testing three beam prisms with dimensions of 40 mm × 40 mm × 160 mm according to the ASTM C 348-18 standard on day 28.The thermal conductivity tests were performed on the flexural test specimens. The samples were heated for 48 h in an oven at a temperature of 60 °C and periodic weight measurements were taken to insure there was no change in the sample weight and thus has reached steady-state; indicated there was no more moisture remaining within the samples that may affect measurements ,32. The The microstructure of a material influences its physical, mechanical, and thermal properties ,36,37. IIn this work, µCT images of concrete samples selected for microstructural investigations were captured using a Thermo-Fisher HeliScan   micro computed tomographic scanner. The 25.4 mm diameter core samples were scanned using the maximum field of view of the system using space filling and large X-ray spot size modes, achieving a voxel resolution of 10.8 µm. The scanning procedure used an X-ray tube potential of 120 kV with a target current of 55 mA. A 0.1 mm steel shim filter was used on the X-ray tube diamond window to suppress beam hardening effects, with aluminum filters ranging between 4 mm (Mixture 6) and 6 mm (Mixture 1) thickness installed on the instrument’s flat panel detector. Samples were heated for 48 h in an oven at 60 °C to ensure that no moisture remained in matrix prior to scanning. Samples were then cooled to room temperature prior to scanning, to mitigate sample movement due to thermal contraction during image acquisition.Microstructural analysis of the CT scans was performed using Thermo-Fisher Avizo Fire  image analysis suite. In order to study the effect of EP addition in the concrete mixture upon concrete microstructure, the volume fractions of the cementitious material paste, air voids, and EP were measured. To reduce computational overhead, each concrete sample volume separated into 14 sub-volumes, each consisting of ~160 orthoslices .The image analysis workflow was conducted separately on each block. Interactive thresholding was implemented to ensure clean segmentation between the paste material, the observable pore-space and EP content. Interactive thresholding is a gradient cut method, whereby the user visually determines grayscale cut off values to segment each material phase. After quality checking the results of the segmentation, quantitative image analysis was conducted to measure the amount of paste to air within each of the imaged concrete mixtures.The imaged samples containing expanded perlite fragments offer considerable challenges in terms of macro-pore characterization using µCT imaging, with EP particulate having similar X-ray attenuation characteristics  to air. In this work, blob detection, and sphericity analysis is used to segregate the two aforementioned material phases from µCT images. Sphericity is the degree to which an object conforms to a perfect sphere. Qualitative analyses of the control concrete mix (mixture 1) showed that the air voids exhibit highly were spherical geometries, with EP typically exhibiting a high degree of angularity and low sphericity, providing a practical criteria through which to segment the phases. To achieve this objective; label analysis was performed on the EP-air void segmented binary images to separate the blobs via their connected components. The sphericity of each disconnected blob was calculated ashericity . The vol2 values. For discussion purposes, the mixture with 0% EP is referred to as the reference mixture. The highest compressive strength, 91.4 MPa, was observed for the reference mixture on day 56. Two main observations were noted. First, as expected, the compressive strength of the concrete material decreased as the amount of EP present in the mixture increased. In particular, the largest decrease in compressive strength with respect to the reference mixture was noted at 20% and 40% EP. However, the decrease in compressive strength when the EP content increased from 60% to 80% and from 80% to 100% was not as significant when tested on days 28 and 56.The compressive strength testing results for each concrete mixture at days 1, 7, 28, and 56 are given in The decrease in compressive strength with increasing amounts of EP in the concrete mix can be explained by the low compressive strength of EP owing to its high porosity; therefore, as more amounts of sand are replaced by EP, the compressive strength of the mixture inherently decreases.2) values, which indicate good exponential fitting to the data points. The error bars in the graph represent the standard deviation between the results for each reported average.Despite this decrease in compressive strength, the resulting measurements are still acceptable and are comparable to the strengths of structural concrete mixtures. 2 of 0.80. The flexural strength of the concrete decreased from 14.2 MPa at 0% EP in the mixture to 10.6 MPa when 100% of the sand was replaced by EP.−1·K−1 at 0% EP to 0.69 W·m−1·K−1 at 100% incorporation of EP; thus, achieving a desirable 62% decrease in thermal conductivity.The thermal conductivity of a material is a measure of its ability to transfer heat. Typically, materials with higher levels of thermal conductivity  are efficient for conducting heat, whereas the opposite is true for materials that exhibit lower levels of thermal conductivity . Porosity is a main factor that affects the thermal conductivity of concrete because the pores are filled with air, which has low thermal conductivity. The replacement of normal aggregate by EP increases the total porosity of the concrete and thus decreases the thermal conductivity and increases the thermal insulation of the concrete. This is illustrated in In this section, μCT analysis is performed for three of the mixtures, namely, mixtures 1, 4, and 6. Mixture 1 (0% incorporation of EP) was chosen as it is the baseline for comparing the other mixtures that incorporate EP. Mixtures 4 and 6 were chosen to demonstrate the properties of the concrete when 60% of the aggregate is replaced with EP (mixture 4) and when 100% of aggregate is replaced with EP (mixture 6). Interactive thresholding was applied to the images to segment the image into paste and air voids/EP particles. To segregate the air voids and EP in mixtures 4 and 6, with 60% and 100% EP incorporation, respectively, a geometric segmentation based upon sphericity was used. More specifically, image features with sphericity values equal to or less than 0.78 were labeled as EP, and image features with sphericity values over 0.78 were labelled as air voids. Lightweight concrete may experience segregation between the aggregate and the paste material ,40,41, tTo corroborate results of the µCT image analysis presented herein, the data in The replacement of normal aggregate (sand) by EP in concrete mixtures exhibited favorable properties, particularly in the significant decreases in thermal conductivity. Although the incorporation of EP did decrease the overall compressive and flexural strengths of the concrete, the decrease was not significant, and the resulting measurements were comparable to those reported in the literature and to measured values for standard industry-grade concrete. Therefore, the use of concrete containing EP is justifiable for structural elements. The work in  investigThis paper introduces a lightweight but strong concrete mixture that uses EP to replace varying volumes of sand aggregate. The proposed mixture obtained superior strength in comparison with previous studies due to introducing perlite to a reference/control mix that had high compressive strength (approximately 80 MPa at 28 days). This strength was achieved primarily by using low W/C ratio and different admixtures especially micro silica.The proposed mixtures showed high compressive and flexural strengths, making it suitable for use in appropriate structural elements with similar strength requirements. In addition, the low thermal conductivity of the proposed concrete makes it a promising material for use in buildings because the improved thermal insulation results in lower energy consumption for heating and cooling. Furthermore, the microstructural analysis via CT imaging revealed that the proposed lightweight concrete mixtures are relatively homogenous throughout the samples, which is critically important but uncommon in lightweight concrete.Future research will focus on conducting life cycle assessment and life cycle costing to determine the sustainability benefits of using the EP mixtures developed in this study in various structural elements and buildings.


Bacteriophage (phage) therapy is a promising approach to combat the rise of multidrug-resistant bacteria. Currently, the preferred clinical modality is to pair phage with an antibiotic, a practice thought to improve efficacy. However, antagonism between phage and antibiotics has been reported, the choice of phage and antibiotic is not often empirically determined, and the effect of the host factors on the effectiveness is unknown. Here, we interrogate phage-antibiotic interactions across antibiotics with different mechanisms of action. Our results suggest that phage can lower the working MIC for bacterial strains already resistant to the antibiotic, is dependent on the antibiotic class and stoichiometry of the pairing, and is dramatically influenced by the host microenvironment. Escherichia coli (ExPEC) with antibiotic levels blanketing the MIC across seven orders of viral titers. Our results suggest that, under certain conditions, phages provide an adjuvating effect by lowering the MIC for drug-resistant strains. Furthermore, synergistic and antagonistic interactions are highly dependent on the mechanism of bacterial inhibition by the class of antibiotic paired to the phage, and when synergism is observed, it suppresses the emergence of resistant cells. Host conditions that simulate the infection environment, including serum and urine, suppress PAS in a bacterial growth-dependent manner. Lastly, two different related phages that differed in their burst sizes produced drastically different synograms. Collectively, these data suggest lytic phages can resuscitate an ineffective antibiotic for previously resistant bacteria while also synergizing with antibiotics in a class-dependent manner, processes that may be dampened by lower bacterial growth rates found in host environments.The continued rise in antibiotic resistance is precipitating a medical crisis. Bacteriophage (phage) has been hailed as one possible therapeutic option to augment the efficacy of antibiotics. However, only a few studies have addressed the synergistic relationship between phage and antibiotics. Here, we report a comprehensive analysis of phage-antibiotic interaction that evaluates synergism, additivism, and antagonism for all classes of antibiotics across clinically achievable stoichiometries. We combined an optically based real-time microtiter plate readout with a matrix-like heat map of treatment potencies to measure phage and antibiotic synergy (PAS), a process we term synography. Phage-antibiotic synography was performed against a pandemic drug-resistant clonal group of extraintestinal pathogenic  A major public health crisis is the alarming increase of infections caused by antibiotic-resistant bacteria, which are responsible for approximately 2.8 million infections in the United States alone . In 2014extraintestinal pathogenic Escherichia coli (ExPEC) with the sequence type 131 (ST131)  . This gr (ST131) , 6. In a (ST131) .31 total particles)  . The adarticles) , 10. Pharticles) . Finallyrticles) . These cin vitro assessments of phage-antibiotic action prior to treatment to select for synergistic combinations, a personalized approach that has led to satisfactory therapeutic outcomes in those cases E. coli phages in our library that targeted JJ2528, but ΦHP3 was shown to have very good killing against JJ2528 both in vitro and in a mouse model of bacteremia  in vitro, can circulate to major organ tissues  in mice, and was able to reduce disease severity in mice for the bacteremia model . This way, multiples of the Sn10  can be used as a practical parameter when defining the concentration of each for an assay, experiment, or treatment.Throughout the study, we found that synograms seemed to follow a pattern specific to the antibiotic being tested and were generally divided into three sections: (i) an antibiotic-dominated-killing region, usually the upper division of the synograms, where antibiotics are effective and thus the killing pattern of the combination therapy closely tracks that of the antibiotic-only treated cells  when bacterial cells were treated with ≥32 μg/ml of chloramphenicol, a region that is predominantly affected by the action of the antibiotic . CAT functions to transfer an acetyl group from coenzyme A to chloramphenicol, a modification that prevents the antibiotic from binding to the bacterial ribosome, prohibiting the inhibition of protein synthesis. We first examined the effect of each agent alone and in combination on wild-type JJ2528 that lacks the On the other hand, ExPEC harboring the enzyme CAT (JJ2528-CAT) was highly resistant to chloramphenicol; it took >32-fold more  chloramphenicol to achieve a similar reduction as the wild-type JJ2528 that lacks the enzyme . Once ag10.1128/mBio.01462-20.2FIG S1P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. Download FIG S1, TIF file, 2.2 MB.Violin plots for the interacting region of the synograms. Areas under the curves (AUCs) for the possible interacting regions of the synograms are shown in violin plots where the medians are the solid thick vertical lines and the quartiles are solid thin lines. (A) Comparison between bacterial resistance; (B) comparison between classes of antibiotics; (C) comparison between media. AUCs were normalized. Kruskal-Wallis test was performed, followed by Dunn’s test for multiple comparisons. *, Copyright © 2020 Gu Liu et al.2020Gu Liu et al.Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the The acquisition of a gene (such as that above) that encodes an enzyme or other protein that inactivates, blocks, or pumps out the antibiotic is one mode by which bacteria become resistant to antibiotics. Another mode, however, is that the gene acquires mutations that enhance the corresponding enzyme’s catalytic activity. To determine the effect of phage-antibiotic combinatorial treatment on bacteria harboring such changes, we introduced genes encoding the β-lactamases CTX-M-14 wild type (WT) and CTX-M-14 A77V/D240G into JJ2528. CTX is an enzyme that hydrolyzes the β-lactam ceftazidime; the presence of the double mutations (A77V/D240G) allows the bacteria to hydrolyze ceftazidime more efficiently than the wild-type enzyme . Mutant 28–4 PFU/ml) or phage-antibiotic-treated cells (7 to 109 PFU/ml of ΦHP3) was not statistically significant  showed dramatically different interactions with the same phage over the same concentration of both agents suggested that the outcome of phage-antibiotic interactions might vary in a consistent manner by antibiotic class (or mechanism of action). To test this hypothesis, we determined the effect on JJ2528 when ΦHP3 was combined with representatives of four other classes of antibiotics, including trimethoprim (folic acid synthesis inhibitor), colistin (cell membrane disrupter), ciprofloxacin (DNA topoisomerases inhibitor), and kanamycin (protein synthesis inhibitor-30S subunit). Wild-type JJ2528 treated with ΦHP3 and the folic acid synthesis inhibitor trimethoprim produced a synogram dominated by phage killing . In facted cells . Of notenificant . This ty4 PFU/ml , which led to 73% reduction. However, this seemingly effective combination was again reversed when the ciprofloxacin doubled to 16 μg/ml, which in combination with 108 PFU/ml of ΦHP3, led to only 11% reduction  was applied as a single treatment  and human heat-inactivated serum . Note that even with additional heat inactivation, untreated bacterial levels decreased over time, especially >8 h; hence, we analyzed the serum synogram using the 8th hour as the endpoint. We first chose an antibiotic that synergizes well with ΦHP3 against wild-type JJ2528 in LB, which was the β-lactam ceftazidime . InteresTo determine if the types of PAS observed to this point are affected by the choice of phage, we chose to perform synography with phage that is 98% identical to ΦHP3 (termed ΦES12) but harbors a 6-fold reduced burst size (60 PFU/cell for ΦHP3 compared to 10 PFU/cell for ΦES12). We examined synography when ΦES12 was combined with ceftazidime (synergism previously observed with ΦHP3) and ciprofloxacin (antagonism previously observed with ΦHP3). Unexpectedly, the combination of ΦES12 and ceftazidime resulted in a synogram with mostly additive effects and a few synergistic combinations, with the overall patterning very different in composition compare  with 6B.in vivo efficacy of the antimicrobials, improve clinical outcomes, and decrease resistance, we tested a recently discussed phage highly specific against the pandemic ST131 clonal group of E. coli for synergistic interactions with all major classes of antibiotics. Primary findings from our study reveal (i) the development of a new high-throughput platform that quickly assesses the effect of various phage and antibiotic concentrations on bacterial growth, an analysis we call synography (and the resulting data represented in a synogram); (ii) that synograms demonstrate a wide range of conditions in which combinatorial treatment is synergistic, additive, or antagonistic, sometimes all three present in the same analysis; (iii) that a phage may demonstrate highly effective killing or inhibition when combined with one class of antibiotics but may lack this same effect when combined with another class; (iv) that phage may restore the competency of antibiotics even in bacteria that encode resistance elements against the chosen antibiotic, an effect we term “phage adjuvation” because the phage adjuvates, or make better, the antibiotic; (v) that highly genetically similar phages produce dramatically different synograms even when these are combined with the same class of antibiotics; (vi) that phage-antibiotic synergy may prevent resistance, but only when the antibiotic concentration is increased; and finally, (vii) the host-like conditions substantially influence PAS, and the synogram profiles in general, thus reflecting the need to test such antibacterial effects under conditions that more reliably simulate the host environment. In this case, the dampening of PAS seems to be due to a reduced growth rate when the bacterium is in urine or blood.Recognizing that a phage used in combination with antibiotics might yield possible beneficial interactions that can enhance An area of phage therapy that is in its infancy is whether phage can help resensitize resistant bacteria toward antibiotics. Often, the options for antibiotics can be limited by bacterial resistance or by the patient’s own medical conditions. In this investigation, we studied the efficacy of combinatorial therapy using antibiotics that can be targeted by bacterial enzymes; these antibiotics are rendered ineffective if they were to be used alone. We found that the MIC was lowered for both wild-type and resistant JJ2528 when the antibiotic was combined with phage. In other words, phages seemed to act as a type of adjuvant to antibiotics, much like we consider alum for antigens in vaccines, even with bacteria harboring genetic elements that confer resistance to the antibiotic. In this sense, they “resensitize” bacteria by lowering the dose of antibiotic needed to achieve a similar reduction of bacterial levels compared to that for the antibiotic alone. There are other ways in which bacteria can be resensitized, for instance, through transduction whereby sensitizing genes are delivered to bacteria after phage infection . As notein vitro  observed additivism, synergism, antagonism, and neutrality; (ii) found that synergistic effects with an antibiotic do not translate to a different antibiotic of the same class that targets similar bacterial pathways; and (iii) showed that more than one type of interaction can exist simultaneously. Since there is a myriad of interactions between phage and antibiotic, there lies a possibility that the interference of some bacterial processes by certain antibiotics may also affect the lytic cycle of the phage. For instance, the synergism and antagonism seen in the membrane disrupter colistin can be caused either by the complexity of the drug (major and minor forms with amphiphilic property) or by its primary interaction with lipopolysaccharide (LPS) that leads to cell membrane destabilization , 33. Sin36–in vitro , 43. Thuin vitro . The bacin vitro . Since pin vitro , 47.From an evolutionary standpoint, the imposition of two different selective pressures on bacteria may reduce the chances for them to develop potential resistance . In our 49–in vitro testing may not always correlate to in vivo efficacy. An inoculum effect was discovered to be an in vivo concern, and this represents only one of the tightly regulated parameters  chloramphenicol on JJ2528-WT and JJ2528-CAT; (B) ceftazidime on wild-type JJ2528, JJ2528 CTX-M-14 wild-type, and JJ2528 CTX-M-14 A77V/D240G. Inoculum effect was also tested on JJ2528 wild-type with different antibiotics: (C) trimethoprim; (D) colistin; (E) kanamycin; (F) ciprofloxacin. Dashed lines represent the limits of detection. Download FIG S2, TIF file, 1.8 MB.Inoculum effects of different antibiotics. A 100-fold diluted subculture of ExPEC (strain JJ2528) was incubated for 4 h, centrifuged, washed, adjusted to an ODCopyright © 2020 Gu Liu et al.2020Gu Liu et al.Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Phages that are chosen for therapy should be carefully characterized to avoid the presence of potentially harmful genes that yield toxins, antibiotic resistance, and virulence to the bacterial host. From our recent characterized phage library , we chosOverall, our investigation of phage-antibiotic interactions paves a new path to explore the complexity of success of dual therapy, especially under physiological conditions. The approach taken in this study can be used to inform clinicians on the possible antagonism that may arise during PAS testing. The real value of the findings reported here is they reveal that the phage-antibiotic interaction is quite complex and influenced by many factors that have not readily been systematically studied. Our work demonstrates (i) how different interactions between phage and antibiotic are strongly affected by the class of antibiotics, (ii) how phage generally lowers the MIC of the antibiotic, (iii) how phage and antibiotics suppress resistance, (iv) how bacterial resistance toward antibiotics impacts the combination therapy, (v) how host factors such as urine and serum affect these types of interactions, and (vi) how similar phages may result in dramatically different outcomes. For future studies, synograms can be modified to accommodate the combination of a phage cocktail combined to an antibiotic or a phage combined to multiple antibiotics. In addition, determining the efficacy of a phage cocktail combined with antibiotics in complex host systems, the effect of simultaneous versus sequential treatment to reduce antagonistic interactions, and the actual mechanisms behind each synergistic and antagonistic effect in combinatorial treatment are fertile grounds for future research.KY608967) and ΦES12 (accession number MN508614), were previously isolated from environmental sources , and 100 μl was inoculated into each well of the microtiter plate that contained the checkerboard of phage and antibiotic concentrations . The OD600 was measured every 15 min at 37°C for a total of 24 h with continuous shaking in a Biotek Synergy HT . For more details on synergy assays, urine collection, and serum information, see Synergy testing was performed with LB medium, human pooled urine, and commercial human serum. A subculture of To determine the effect of inoculum size on the efficacy of antibiotics, 2-fold serial dilutions of antibiotic were added to a 96-well plate. A bacterial subculture of JJ2528 was prepared as described for synergy testing, and serial 10-fold dilutions were inoculated into the microtiter plate and grown for 24 h at 37°C in a shaker. For each bacterial inoculum, the well with the lowest antibiotic concentration that showed bacterial clearance was marked as the MIC. For more details on antibiotics and bacterial dilution, see growthcontrol − ODtreatment)/ODgrowthcontrol] × 100.To generate synograms, absorbance readings from three biological replicates were normalized with the negative control, and the treated wells were deducted from the positive control (no treatment) to yield percent reduction: Reduction (%) =  × 100. The regions above the dashed lines indicate antibiotic-mediated killing with highly effective doses; the regions between the solid and dashed lines represent the interacting regions of the phage and antibiotic, and the regions below the solid lines indicate phage-mediated killing with ineffective antibiotic concentrations. Download FIG S3, TIF file, 1.1 MB.Synograms of different time points for ceftazidime and ciprofloxacin. A 100-fold diluted subculture of wild-type JJ2528 was incubated for 4 h, centrifuged, washed, adjusted to an ODCopyright © 2020 Gu Liu et al.2020Gu Liu et al.Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the 10.1128/mBio.01462-20.5FIG S4600 of 1, and inoculated in a 96-well plate coated with ΦHP3 and antibiotics, and the OD600 was measured every 15 min for a total of 24 h with shaking. Synograms (t = 24 h) were computed for the following classes: (A) cell wall synthesis inhibitors (ceftazidime and cefepime); (B) DNA topoisomerases inhibitors (ciprofloxacin and levofloxacin); (C) cell membrane disrupters (polymyxin E and polymyxin B); (D) folic acid synthesis inhibitors (trimethoprim and sulfamethoxazole); (E) protein synthesis inhibitors (kanamycin and chloramphenicol). Synograms (t = 24 h) represent the mean reduction percentage of each treatment from at least two biological replicates: Reduction (%) = [(ODgrowthcontrol − ODtreatment)/ODgrowthcontrol] × 100. The regions above the dashed lines indicate antibiotic-mediated killing with highly effective doses; the regions between the solid and dashed lines represent the interacting regions of the phage and antibiotic, and the regions below the solid lines indicate phage-mediated killing with ineffective antibiotic concentrations. Download FIG S4, TIF file, 1.1 MB.Synograms of antibiotics with similar mechanisms of action. A 100-fold diluted subculture of wild-type JJ2528 was incubated for 4 h, centrifuged, washed, adjusted to an ODCopyright © 2020 Gu Liu et al.2020Gu Liu et al.Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the 10.1128/mBio.01462-20.6FIG S5600 of 1.0, and inoculated in a 96-well plate coated with different antibiotic concentrations and phage titers. The plate shows the raw absorbance readings for one biological replicate. Download FIG S5, TIF file, 1.2 MB.Diagram of a 96-well plate showing raw absorbance readings for the combinatorial treatment on wild-type JJ2528. A 100-fold diluted subculture of ExPEC (strain JJ2528) was incubated for 4 h, centrifuged, washed, adjusted to an ODCopyright © 2020 Gu Liu et al.2020Gu Liu et al.Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the


Autopsy tissues  served as controls. All sections were studied histopathologically. (3) Results: Pathomorphology in TAAD showed predominantly moderate elastic fiber fragmentation/loss, elastic fiber thinning, elastic fiber degeneration, mucoid extracellular matrix accumulation, smooth muscle cell nuclei loss, and overall medial degeneration. The control group showed significantly fewer signs of those histopathological features . It was concluded that the dissection plane consistently coincides with the vasa vasorum network, and that TAAD associates with a significantly thinner intimal layer p = 0.005). (4) Conclusions: On the basis of the systematic evaluation and the consistent presence of diffuse, pre-existing medial defects, we hypothesize that TAAD relates to a developmental defect of the ascending aorta and is caused by a triple-hit mechanism that involves (I) an intimal tear; and (II) a diseased media, which allows (III) propagation of the tear towards the plane of the vasa vasorum where the dissection further progresses.(1) Background: The pathophysiologic basis of an acute type A aortic dissection (TAAD) is largely unknown. In an effort to evaluate vessel wall defects, we systematically studied aortic specimens in TAAD patients. (2) Methods: Ascending aortic wall specimens ( An acute aortic dissection is a medical catastrophe  with an An aortic dissection is hallmarked by a tear in the intimal layer, which allows blood to access the middle layer of the wall, causing the vascular layers to separate (dissect) from one another, resulting in formation of a ‘’false’’ lumen. One or multiple re-entry tears down- or upstream allow flow in the false lumen. Pressure build-up in the false lumen may result in compression of the aortic side branches. Depending on the degree of occlusion, and the side branches involved, this may result in life-threatening ischemia. The large majority of dissections are continuous with the actual arterial lumen via one or more intimal ‘’entry tears’’, yet some dissections may present without an apparent continuity with the actual lumen . Based oThe pathophysiologic basis of TAAD is poorly understood. Although the disease is associated with hypertension ,2,3, theA role for (an) underlying aortic defect(s) as predisposing factor for TAAD is further supported by a number of histological evaluations ,7,8. UnfWith the aim of harmonizing the histomorphological characterization of degenerative vascular pathologies, the Society for Cardiovascular Pathology along with the Association for European Cardiovascular Pathology recently issued a consensus statements with an unambiguous nomenclature . This noOur study complies with the Declaration of Helsinki. Approval for this study was granted by the Medial Ethical Committee of Leiden University Medical Center (LUMC), Leiden. All control specimens were obtained post mortem. Obduction has been performed according the guidelines of the pathology department. Tissue collection was performed according to the regulations and protocols for secondary tissue use of the dept. of pathology at the Leiden University Medical Center. We have obtained permission to include the specimens in our study and obduction. Ascending aortic wall samples were obtained from the aortotomy site as residual aortic wall material during an ascending aortic replacement in patients with acute TAAD. Seventeen control aortas were obtained during post-mortem autopsies. The wall samples were formalin-fixed, decalcified using Kristensens solution, and paraffin-embedded. Successive 4 µm sections were, respectively, Haematoxylin-Eosin (HE)-, Resorcin-Fuchsin (RF)-, and MOVAT-pentachrome-stained. Vasa vasorum were visualized by CD31 staining , 1/1000 after Tris EDTA  heat retrieval. The anti-mouse Envision+ System HRP  and DAB solution  were used for visualization.Histomorphological grading was performed according to the consensus classification for degenerative aorta pathology . ElasticThe relative position of the dissection plane in the media was estimated as the ratio of the distance from the first major internal elastic lamella (lamina elastica interna) to the dissection plane over the full medial thickness  . For thet-test was used to compare the ratio of the dissection plane to ratio of the vasa vasorum plexus.Statistical analysis was performed using the IBM SPSS statistics software, version 26. Chi-square test was used to compare the morphology between the dissection and the control group. Independent samples This study includes 58 successive TAAD patients and 17 control patients. Baseline characteristics of the patients are shown in Putative aortic wall abnormalities in TAAD were mapped according to the revised, consensus nomenclature for noninflammatory, degenerative aortic histopathologies . Conclusp = 0.044)  vs. 193 ± 132 μm (mean ± sd) in the control group (= 0.044) . Intimaln = 25).From the histological evaluation, the picture emerged of a uniform dissection plane for all TAAD specimens in the study. We therefore evaluated the level of the dissection plane for the 41 cases for which full-thickness aortic wall samples were available. For the remaining 17 samples, the adventitial aspect of the media was either lost or fully separated from the media in the process of tissue procurement. In the available full thickness samples, the dissection plane was consistently located in the outer aspect of the media (at 83% (SD 0.08) of the full medial thickness), a level that coincides with the aortic vasa vasorum network . The levThis systematic histological evaluation of aorta segments of type A dissections shows universal presence of diffuse, moderately-severe medial defects, a thinner intima, and a dissection plane that is consistently located at the level of the vasa vasorum network in the outer media. On basis of these observations, the picture emerges of aortic dissection being the consequence of a multi-hit process that involves a potentially more vulnerable (thinner) intima, a pre-existing incompetent media that is unable to withstand the tear forces exerted by the blood following an intimal tear, thereby allowing the blood access to an area of least resistance  at the level of the vasa vasorum network in the outer medial layer.Histological evaluation indicated universal presence of medial defects in all TAAD cases included in this evaluation. Medial defects were systematically inventoried by applying the recently issued consensus grading from the Society for Cardiovascular Pathology and the Association For European Cardiovascular Pathology . This coThis is the first study to apply the consensus classification to TAAD wall samples. Based on this grading, it was concluded that TAAD universally associates with moderately severe elastic fiber pathology, overall medial degeneration, mucoid extracellular matrix accumulation, and smooth muscle cell nuclei loss. Additionally, MOVAT staining implied further pathological changes not included in the consensus classification  such as Apart from the consistent presence of the moderately severe medial defects, this systematic histological inventory also revealed a remarkably thin intima, and the existence of uniform dissection plane, as consistent findings in the 58 TAAD cases.Although one could speculate that the observed thin intima in TAAD constitutes the primary defect, with thin intima’s being more prone to dissection and the medial defects reflecting a bystander phenomenon, such a theory is not compliant with the clinical observations that intimal tears or medial injuries in a healthy vessel do not result in dissection. Localized intimal tears are observed as incidental findings in elderly patients or after trauma . SimilarAn alternative explanation for the thin intima is that it reflects an underlying developmental and/or differentiation defect of the mesenchymal cell population . RelativA further remarkable observation was the uniform localization of the dissection plane  ,24. The One could speculate that the level harboring this vascular network structure is an inherently mechanically weak plane that is less able to resist the wedge forces exhibited by the blood than the other segments of the media, thereby creating an inherently weak plane within the aortic wall along which the dissection progresses. Considering the fact that the mean blood pressure in the vasa vasorum network is lower than the central blood pressure, we consider it unlikely that TAADs that present without an apparent continuity with the actual lumen relate to a bleeding from the vasa vasorum plexus.In conclusion: this systematic histological evaluation of 58 successive TAAD shows that TAAD uniformly associates with pre-existing ascending aortic wall defects and consistently proceeds along a plane that coincides with the vascular plexus in the outer media. On this basis, a picture emerges of a TAAD-susceptible phenotype that relates to an inherently mechanically incompetent media, and a common final pathway reflecting propagation of the dissection along the plane of least resistance (the vasa vasorum network).This is an observational study. As such, the study comes with several limitations: the evaluation is based on histological evaluation surgical specimens obtained at the time of aorta replacement, hence information on non-operated patients is missing. Moreover, information on the natural history of the medial changes, and on the molecular aspects of TAAD, is missing.


Misuse and overuse of antibiotics by physicians in the treatment of children is common in China. This study aimed to reveal the overall use of antibiotics to treat children hospitalized in four types of pediatric wards.Seven independent point prevalence surveys (PPSs) were conducted in Shanghai Children’s Hospital of Shanghai Jiao Tong University over the period 2012 to 2018. Pediatric ward types were defined general pediatric medical, pediatric surgical, pediatric intensive care units (PICU), and neonatal.n = 1743), penicillins , carbapenems , nitroimidazoles  and macrolides . The five most commonly used generic antibiotics were cefuroxime , ceftriaxone , cefotaxime , meropenem  and ampicillin/sulbactam . Meropenem was among top five antibiotics prescribed in the general pediatric, PICU and neonatal wards and sixth in the pediatric surgical wards. Of all children on antibiotics, 23.4% received prophylactic treatment, and prophylaxis accounted for 68.1% of indications for treatment in the pediatric surgical wards.A total of 3975 pediatric patients were included in the study, of which 63.9% received at least one dose antibiotic. The top five classes of antibiotics administered were cephalosporins (43.8%, Given that over-treatment with third-generation cephalosporins and carbapenems has been associated with treatment-resistant infections, the prescription of these drugs should be strictly controlled and monitored, and measures should be taken to improve the management of surgical prophylaxis in hospitalized children in China. China has the largest population in the world and also uses a large quantity of antibiotics. A study conducted in 784 community health institutions in 28 cities across China showed that China’s community health institutions prescribed many more antibiotics and more injections than the WHO’s recommendations . ChildreMisuse and overuse of antibiotics causes not only the most common side effects such as antibiotic-associated diarrhea and antibiotic allergic reactions but also higher healthcare costs . If the This study analyzes the results of seven repeated point prevalence surveys (PPSs) on antibiotic usage carried out in Shanghai Children’s Hospital from 2012 to 2018. We aimed to reveal the general situation of antibiotic prescription practices, including antibiotic number, type of antibiotic agents, antibiotic prescription rates and the indications for antibiotic treatment  among hospitalized children in four main types of pediatric wards.The study was conducted in Shanghai Children’s Hospital of Shanghai Jiao Tong University, a tertiary level 800-bed hospital with all medical specialties available, including PICU, neonatal intensive care unit (NICU), neonatology, hematology oncology, respiratory, gastroenterology, nephrology, endocrinology, cardiology, neurology, neurosurgery, cardiothoracic surgery, general surgery, urology, pediatric orthopedics, otolaryngology, and ophthalmology. Data was collected from all wards of the hospital.A one-day PPS on antibiotic treatment of hospitalized children was performed in Shanghai Children’s Hospital. A total of seven PPSs were conducted during the period 2012 to 2018 (once a year in the months of November or December). The survey included all admitted patients present in the ward at 8:00 am on the day of the survey. Patients discharged on the survey day were included, and newly admitted patients were excluded. The total number of patients included in the study was 3975. The number of participants per survey day was as follows: 376 (2012-11-13), 399 (2013-11-20), 578 (2014-11-12), 643 (2015-12-10), 575 (2016-11-30), 684 (2017-12-12) and 720 (2018-12-11). One pediatrician and one nurse in each ward were responsible for investigating and registering the use of antibiotics in the hospitalized children. For all participants, data about gender, age, clinical department, disease diagnoses, and antibiotic treatment characteristics including the categories of antibiotics prescribed, the reasons for prescribing and combinations antibacterial medication were collected. In addition, surgical inpatient records included the surgical site and time of surgery. The study was approved by the Research Ethics Committee of Shanghai Children’s Hospital of Shanghai Jiao Tong University . Our team has obtained administrative permissions to access the data used in the study.P value < 0.05 was considered to be statistically significant.Microsoft Excel 2007  and SPSS 16.0 statistical package  were used. The categorical variables were presented as counts and percentages. Statistical significance for the differences between groups was assessed using a chi-square test and a two-sided n = 1514), pediatric surgical , PICU  and neonatal wards . Neonatal wards included the NICU, premature, and a general neonatal medical wards. The demographic characteristics and disease diagnoses of the patients included in the current study are summarized in Table A total of 3975 patients (1546 girls and 2429 boys) were hospitalized during the seven PPSs. The median age of the cohort was 2  years, with a minimum of 1 h and a maximum age of 17 years. Patients were in four main pediatric ward types: general pediatric medical , penicillin-based drugs , carbapenems , nitroimidazoles  and macrolides . Among cephalosporins, third-generation drugs represented 59.4% (n = 1035) of prescriptions, while the second-generation drugs accounted for 40.3% (n = 702) of prescriptions. The combination of penicillins plus β-lactamase inhibitors accounted for 86.69% (456/526) of penicillin-based prescriptions. Carbapenems were the third most frequently prescribed antibiotic class in our study, with meropenem the most frequently used in this class. The five most commonly used generic antibiotics were cefuroxime , ceftriaxone , cefotaxime , meropenem  and ampicillin/sulbactam  , carbapenems, nitroimidazoles, macrolides, glycopeptides, lincosamides, cephalomycins, sulfonamides, linezolid, aminoglycosides, fosfomycin, furans, tetracyclines, glycylcyclines, quinolones, aztreonam, and antifungals. Overall, 2541 (63.9%) children received at least one dose of an antibiotic during the seven PPSs. The top five classes of drugs prescribed were cephalosporins , the pediatric surgical wards , the PICU  and the neonatal wards ; . There was no significant difference of antibiotic use rate between boys and girls in the four types of wards . In the general pediatric medical wards, 40 different antibiotics were recorded of which cefuroxime was the most often prescribed  followed by ceftriaxone , azithromycin , amoxicillin/clavulanic acid  and meropenem . In pediatric surgical wards, 34 different antibiotics were recorded, of which cefuroxime was the most often prescribed  followed by ornidazole , ceftriaxone , cefoperazone/sulbactam  and metronidazole . In the treatment of children in PICU, 30 different antibiotics were recorded, of which meropenem was the most often prescribed  followed by cefoperazone/sulbactam , cefuroxime , ampicillin/sulbactam  and azithromycin . In neonatal wards, 28 different antibiotics were recorded, of which cefotaxime was the most often prescribed  followed by ampicillin/sulbactam , meropenem , ticarcillin/clavulanic acid  and metronidazole  , ornidazole , ceftriaxone , metronidazole  and cefoperazone/sulbactam . The five most commonly used generic antibiotics for therapy were cefotaxime , ceftriaxone , cefuroxime , meropenem  and azithromycin .Of all children on antibiotics, 74.5% received antibiotics for antimicrobial therapy, 23.4% received antibiotics for prophylactic treatment and 2.1% as prophylaxis plus therapy. The five most commonly used generic antibiotics for prophylaxis were cefuroxime , whereas in the pediatric surgical wards prophylactic antibiotic treatment accounted for 68.1% of prescriptions. In addition, 30% of the children in pediatric surgical wards receiving antibiotics received two or more types of antibiotic prophylaxis. The majority (63.5%) of patients receiving antibiotic treatment in the pediatric surgical wards were prescribed two or more types of antibiotics whereas far fewer (36.4%) received a single antibiotic. One half of children in PICU and approximately 30% in the general pediatric medical and neonatal wards received two or more types of antibiotics   but muchConsistent with most study results , 23–25, In this study, carbapenems ranked third in total antimicrobial treatment of pediatric patients and was almost totally represented by meropenem. Meropenem was among top five antibiotics prescribed in the general pediatric, PICU and neonatal wards and sixth in the pediatric surgical wards. Carbapenems are often considered to be last resort agents reserved for the treatment of infections highly resistant to most antimicrobial treatments. Unfortunately, whether it is in the literature  or in clOf all children on antibiotics in this study, 23.4% received prophylactic treatment, and low prescribing rates of antibiotic prophylaxis were observed in the general pediatric medical (2.3%), PICU (4.2%) and neonatal wards (6.3%). This was a reassuring finding, since prophylactic use of antibiotics to prevent infections is controversial. The results of the current study did not support the administration of antibiotics for surgical prophylaxis as a potential quality indicator. Prophylaxis in pediatric surgical wards, however, accounted for 68.1% of antibiotic prescriptions. This was much higher than the percentage of antibiotic prophylaxis for surgical diseases in the treatment of children in Europe (26.6%) . Also, 3There are several strengths to this study. Firstly, all wards were investigated, so our research population included medical and surgical wards with a broad spectrum of diseases. Furthermore, the wards were divided into pediatric medical, PICU, neonatal wards and pediatric surgical wards, which would help to manage antibiotic use based on the unique challenges in the different wards. This distinction was rare in previous studies. Secondly, seven repeated PPSs were conducted on all wards within the same hospital so that the selection bias would be reduced and the overall prescribing of antibiotics in our hospital would be better reflected. There were also certain limitations in our study. Firstly, one of the drawbacks of PPS is that it cannot be judged whether the purpose of using antibiotics at this point is empirical or targeted medication. Secondly, in this study, all PPSs were conducted in autumn and winter, which could not reflect the fact that antibiotic use may change according to seasons of the year. Thirdly, the study was conducted at a single healthcare center, and the results may or may not be able to be extrapolated to other similar hospitals. Nevertheless, as a tertiary children’s hospital in China, patients come from multiple regions of the country. Thus, the results in this study do reflect the use of antibiotics in the treatment of hospitalized children.This study has revealed the high rate of overall antibiotic use in the treatment of children, and most particularly neonates and children requiring surgery, in a tertiary hospital in Shanghai. The most commonly prescribed antibiotics were third-generation cephalosporins. Meropenem was among top five antibiotics prescribed in the general pediatric, PICU and neonatal wards and sixth in the pediatric surgical wards. The prescription of these drugs in children should be strictly controlled and monitored to effectively limit the development of antimicrobial resistance. Additionally, measures should be taken to improve the management of antibiotics for surgical prophylaxis in children. The CDSS is one tool, already deemed useful in the literature, for the management of antimicrobial treatment in children and adults.


Investigations included the kinetics of hybrid materials formation and selected physical and mechanical properties. The disadvantage of ionogels without the methacryl-POSS modifier is leakage and insufficient mechanical properties. Modifying the thiol-ene matrix by the addition of methacryl-POSS made it possible to obtain non-leaking ionogels with improved mechanical and conductive properties. The steric hindrance of POSS cages and high-density network formation played important roles in ionogel synthesis: decrease of polymerization rate , as well as dimensions of the formed polymer spheres during dispersion polymerization , an increase of glass transition temperature and puncture strength. Hybrid ionogels with high ionic conductivity in the range of 4.0–5.1 mS∙cm−1 with the maximum parameter for 1.5 wt.% addition of the methacryl-POSS were obtained, which can be associated with ion-pair dissociations in ionic liquid clusters caused by methacryl-POSS.The effect of polyhedral oligomeric silsesquioxane (POSS) on the synthesis and properties of hybrid organic–inorganic ionogels was investigated using octakis(methacryloxypropyl) silsesquioxane (methacryl-POSS). Ionogels were prepared in situ by thiol-ene photopolymerization of triallyl isocyanurate with pentaerythritol tetrakis(3-mercaptopropionate) in a mixture of imidazolium ionic liquid 1-ethyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide (EMImNTf Hybrid organic–inorganic ionogels are considered to be a new class of hybrid material, containing solid matrix (organic–inorganic nanoparticles in the polymer) and physically entrapped (immobilized) ionic liquid (IL) that keep their unique properties. These materials can be used as membranes or gel polymer electrolytes (GPEs) to produce electrochemical devices . Hybrid ionogels can be obtained by an in situ process based on the polymerization of monomers and fillers dissolved/dispersed in ILs. Classical radical photopolymerization involving (meth)acrylates, represents a valuable approach that exhibits unique spatial, temporal control by means of a chain-growth mechanism, low-temperature cure, and energy efficiency. This process is widely used for a range of applications  ,3,4,5,6.The application of thiol-ene reaction effectively combines the classical benefits of click reactions with the advantages of a photoinitiated process, resulting in an excellent method of chemical synthesis and production of tailorable materials ,12. In c1.5)n or Tn (where n is the number of repeat units and T is RSiO1.5) wherein the substituent R can be reactive or nonreactive in the polymerization process [In our continuing investigations on the preparation and properties of photocurable ionogels with a thiol-ene matrix  , we unde process ,14. This process ,16,17. M process ,18, alth process . When PO process .POSS containing thiol groups was introduced into acrylated castor oil to develop a novel photocurable hybrid material with reduced surface energy and better thermal stability . Fang etDespite many works on polymer/POSS materials, only a few papers focused on the kinetics of the photocuring process. Usually, a small amount of POSS can substantially enhance the photopolymerization rate, both in the case of classical radical-based photopolymerization ,22 and t2 in various blended compositions were also studied by Kim et al. [Nanofillers are incorporated into ionogels mainly to improve their thermal and mechanical properties. The detailed review of the fabrication of hybrid materials with special attention paid to polymer-based ionogels is presented in . The incm et al. ,26. GPEs can be also applied in electrochemical capacitors (EC) where plays the role of separator and electrolyte simultaneously. The most important advantages of using GPE are (i) preservation against the electrolyte leakage from ECs even in the case of its damage and (ii) simplification and time saving of ECs fabrication processes. Time saving is particularly noticeable if GPE is produced by the photopolymerization process ,28. HoweThe thiol-methacrylate system uniquely exhibits a mixed step-chain growth polymerization regime that combines the step-growth thiol-ene reaction and chain free-radical polymerization. This provides a simple route to prepare a novel POSS-based polymer. The effect of ILs and POSS on thiol-ene polymerization can differ from that observed for (meth)acrylate polymerization, because of different polymerization mechanism. The incorporation of methacryloxy functionalized-POSS derivatives is expected to enhance the mechanical and conductive properties of thiol–ene-POSS ionogels.In studies presented in this paper to reduce the costs of ionogel, we decided to use for the synthesis a mixture of IL with propylene carbonate instead of the neat IL. As monomers triallyl isocyanurate (TATT) and pentaerythritol tetrakis(3-mercaptopropionate) (PETMP) were used. The obtained poly(TATT + PETMP) ionogel with the solvent mixture has a major disadvantage of leakage. To prevent this problem, we decided to modify the system with the addition of methacryl-POSS, which should also improve the mechanical properties of the ionogel. The aim of the study was therefore the synthesis of hybrid ionogels modified with the addition of POSS by the thiol-ene photopolymerization in the IL + PC mixture. Moreover, the study of the POSS influence on the kinetics of the reaction and the properties of the obtained ionogels.2), purity 99% was delivered by Solvionic . Propylene carbonate (PC) anhydrous, purity 99.7%, was purchased from Sigma–Aldrich . The photoinitiator, 2,2-dimethoxy-2-phenylacetophenone (DMPA) was also supplied by Sigma–Aldrich . Monomers: pentaerythritol tetrakis(3-mercaptopropionate) (PETMP), purity ≥95%, 1,3,5-triallyl-1,3,5-triazine-2,4,6-trione (TATT), purity 98%. Monomers were delivered by Sigma–Aldrich . Octakis(methacryloxypropyl) silsesquioxane , was provided by Hybrid Plastics . 1-ethyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide . The polymerizations were initiated with the UV light . All the experiments were conducted at least in triplicate. The reproducibility of the kinetic results was about ±3%.The kinetics of photopolymerization of the thiol-ene compositions modified with methacryl-POSS (with or without solvents mixture) was determined by using isothermal differential scanning calorimetry . The DSC apparatus was equipped with a lid especially designed for photopolymerization measurements. The 2 mg samples were polymerized in open aluminum pans (diameter 6.6 mm) under isothermal conditions at 25 °C. All reactions were performed under high-purity argon atmosphere , α (hydrogen bond donating ability), and β (hydrogen bond accepting ability). All dyes were delivered by Sigma–Aldrich . Solvatochromic dyes were dissolved in anhydrous methanol in the concentration of 5 × 10−3 M. The dye concentrate was added to the IL or solvent in such an amount that the maximum absorption band was in the range of 0.4 to 0.5. The mixture was homogenized and then, the methanol was evaporated at 40 °C under reduced pressure. The absorption spectrum was measured for each dye by spectrophotometer Jasco UV-530 . Measurements were carried out at room temperature in a quartz cuvette with a light path length of 1 mm in the measuring range 200–800 nm. The solvatochromic parameters were calculated according to Equations (1)–(5) and the methods described in the articles [N)-4-Nitroaniline, and (DN)-N,N-Diethyl-4-nitroaniline.4-Nitroaniline, purity > 99%; rity 90%  were usearticles ,32,33:. Then, the test samples were cut to certain dimensions from the obtained sheets of ionogels.The samples were prepared in a glove box under a pure argon atmosphere. The photocurable composition consisted of a mixture of monomers, methacryl-POSS, solvents, and photoinitiator (0.2 wt.%) was homogenized in an orbital shaker. The obtained homogeneous composition was poured into 0.3 mm thick glass molds and exposed in (room temperature) RT to UV light for 10 min on each side of the mold . Before measurements, synthesized ionogels were treated with methanol and dried in an oven at 35 °C to remove the solvents mixture (IL + PC). The small pieces of dried samples were placed on a stub of metal with adhesive and coated with an ultrathin Gold/Palladium coating, deposited on the sample by low-vacuum sputter coating. Based on obtained photos, the range of diameters (nm) of polymer particles has been measured and means and standard deviations were calculated. To compare the obtained results, the statistical significance of the difference between means has been tested.gT was measured by DSC  under a nitrogen atmosphere with a heating rate of 20 °C∙min−1 in the temperature range from −80 °C to 100 °C. The gT value was determined from the second run of the DSC measurement. For each ionogel, four measurements of DSC were made. Based on the obtained results, the means and standard deviations were calculated. To compare the obtained results, the statistical significance of the difference between means has been tested.Glass transition temperature 2O3 crucible and was analyzed with a heating rate of 10 °C∙min−1 under nitrogen atmosphere (purge of 10 mL∙min−1 of N2 protection gas and 20 mL∙min−1 of N2 sample gas).Thermal resistance of synthesized materials was investigated with TG 209 F3 Tarsus thermogravimetric analyzer  in the temperature range of 30–600 °C. Approximately 10 mg of sample was placed in Al−1). Because different ionogels slightly differ in their thickness, the load was normalized to a uniform thickness (300 µm) to compensate for the influence of thickness on the measurement. For each ionogel, seven measurements of mechanical properties were made. Based on the obtained results, the means and standard deviations were calculated. To compare the obtained results, the statistical significance of the difference between means has been tested.To characterize the mechanical properties of the obtained ionogels, a puncture resistance test was conducted. The measurements were performed with a CT3 Texture Analyzer . A sample with a diameter of 16 mm was cut out from ionogel just after synthesis. Its thickness was measured and then the sample was fixed in a 10 mm hole diameter sample holder and tested for puncture strength using the spherical probe with 2.5 mm radius. During the measurement, the load and distance of the measuring probe were recorded until the sample was punctured (probe displacement rate–0.3 mm∙sσ) was calculated from Equation (6):σ is the ionic conductivity of the ionogel in S∙cm−1, l is the thickness of the ionogel in cm, A is an ionogel surface area in cm2, and Sσ represents the volumetric conductance of the ionogel sample, S.The ionic conductivity of the ionogels was investigated by electrochemical impedance spectroscopy (EIS) in the frequency range from 1 kHz to 1 MHz using the SP-300 potentiostat/galvanostat . The experiment was performed in a two-electrode (stainless steel—316L) type electrochemical vessel at room temperature. The ionic conductivity of the ionogels —90 wt.% of Maxsorb MSP-20X —with carbon black—5 wt.% of C65 —and binder—5 wt.% of PTFE . The electrode components were mixed in deionized water until a homogenous suspension was obtained. Then, the solvent was partially evaporated, and a thin film was prepared by rolling out and calendaring to an average thickness of 200 ± 15 μm. Thereafter, the electrode film was glued on a stainless steel current collector using Acheson electrodag PF-407C . The mass and the diameter of each electrode were equal 13.0 ± 0.3 mg and 12 mm, respectively.® cell , the carbon electrodes were soaked off in a solution of IL with PC, and the EC was assembled in an argon atmosphere. The cells were investigated by cyclic voltammetry (CV) with various scan rates from 5 to 100 mV∙s−1 and up to maximal cell potentials from 1 to 3 V, galvanostatic (0.5 to 4 A∙g−1) charge/discharge with potential limitation (GCPL), charge/discharge at constant power with voltage limitation between 1.5 to 3 V  and electrochemical impedance spectroscopy at OCV , using an SP-300 potentiostat/galvanostat .Before assembling the Swagelok2 with high ionic conductivity (9 mS∙cm−1) and rather low sensitivity to moisture was used. However, considering the high viscosity and the price of ionic liquids, which make them not commonly used as electrolytes, we decided to prepare a mixture of IL with organic solvent—PC. This solvent is commonly used in electrochemistry [2 in PC was X = 0.2 (solution with the highest conductivity) and for ionogel preparation, 70 wt.% of this mixture was used. The introduction of propylene carbonate to the IL caused a significant increase in conductivity (13.8 mS∙cm−1); however, the use of this mixture for the synthesis of the ionogel caused leakage of the obtained material. For these two reasons, leakage and insufficient mechanical properties of the obtained ionogels, we decided to modify the polymer matrix by adding methacryl-POSS to the initial composition. With the addition of methacryl-POSS, an equimolar ratio of thiol (PETMP) to ene (TATT + 8M-POSS) functional groups was maintained, with the calculations assuming that 6 methacryl-POSS groups were involved in the polymerization reaction because of steric hindrance. The formulations of the compositions used for the synthesis of the polymer matrix are shown in 2 and PC) is shown in The thiol-ene system selected for this study was based on multifunctional monomers, i.e., trifunctional ene TATT and tetrafunctional thiol PETMP. Monomers were used in an equimolar ratio of functional groups . Polymerization of these monomers results in a high-density polymer network formed with a rapid reaction. However, the mechanical properties of the obtained polymer are insufficient due to the flexible thioether linkages formed during thiol-ene polymerization ,35. The hemistry ,37. The Polymerization of thiol-ene compositions goes in a step-growth mechanism by hydrothiolation of double bonds. Polymerization is initiated by the decomposition of a photoinitiator and a hydrogen abstraction from a thiol functional group. Then chain propagation occurs in two steps: propagation reaction and chain-transfer reaction, which results in thiol addition across the alkene. This addition/chain-transfer process continues cyclically. The step-growth nature of this reaction requires an equimolar ratio of reactive groups SH:C=C (1:1) to obtain high conversion . When me2 + PC were homogeneous and transparent .Prepared initial compositions of monomers (TATT + PETMP) and modifier (8M-POSS) with mixture of solvents EMImNTfnsparent a. After nsparent b, which 2 + PC) are depicted in pmaxR), time of reaction (maxt), and conversion (Rmp) at pmaxR on the concentration of the 8M-POSS in the investigated systems are shown in Kinetic curves obtained for photopolymerization of composition TATT + PETMP in the presence of 0, 0.5, 1, 1.5, 3, 5, 7, and 9 wt.% of 8M-POSS provided in 70 wt.% of solvents mixture  and hydrogen bond basicity (accepting ability) of the solvent, respectively, and π* describe dipolarity/polarizability of the solvent [β parameter which is connected with additional stabilization of the transition state during the chain-transfer step. On the other hand, the α parameter contributes to the decrease of polymerization rate due to the reduction in stabilization. In our previous study, we used monomer TATT and trimethylolpropane tris(3-mercaptopropionate) TMTMP, which differs only from the PETMP monomer in that it has one thiol group less in its structure. Thus, the influence of used solvents should be similar for the polymerization of these two compositions of monomers. Used solvents EMImNTf2 and PC caused the increase in polymerization rate of the TATT + TMTMP mixture, with the higher pRmax value obtained in PC. The IL EMImNTf2 has β parameter with lower value (0.23) than PC β parameter (0.35), and moreover with higher value of α parameter . Therefore, the polymerization of TATT+TMTMP proceeds with a faster polymerization rate in the PC than in the IL. Thus, we expected in this study an increase in the polymerization rate using a mixture of PC and EMImNTf2 which is characterized by α Kamlet–Taft parameter equal to 0.59 and β parameter equal to 0.30. In addition, as can be seen in 2 and PC.The influence of ILs and non-ionic solvents on the thiol-ene polymerization we explain in our previous reports ,41. The  solvent ,42. The  solvent . The reaSolvent homopolymerization of the neat methacryl-POSS occurs with much lower reaction rate than solvent thiol-ene polymerization of the TATT + PETMP composition . The stemaxt) and the conversion (Rmp) in the maximum reaction rate (pmaxR) to higher values . As mentioned above, polymerization in bulk proceeds with a lower polymerization rate than in solvents mixture. As can be seen from the comparison of the obtained kinetic curves shown in We also performed investigations of the polymerization kinetics of thiol-en-8M-POSS systems in bulk , withoutAs was mentioned above obtained ionogels were opaque, with phase separation. Thus, the polymer matrix morphology of synthesized ionogels, after extraction of solvents, was investigated by SEM to reveal the structure of the polymer matrix. SEM images of polymer matrices are shown in As can be seen in gT of synthesized materials were measured using DSC. In gT temperatures of the polymer matrix, ionogel without modification, and hybrid ionogels are presented. Pure poly(TATT+PETMP) has glass transition above room temperature, i.e., gT = 35.1 °C because two multifunctional monomers are used to matrix synthesis what results in a dense polymer network formation. Glass transition decreases about 20 °C for ionogel with 70 wt.% of EMImNTf2 + PC mixture. Used solvents plasticize polymer matrix. As shown in gT of hybrid ionogels of about 1–3 °C. However, in the studied range of 8M-POSS concentration, considering the results of the statistical analysis for this parameter , and thus the presence of unreacted thiol groups in the matrix structure, may cause a plasticizing effect (decrease of gT). The additional effect can be exerted by the intermolecular interactions of the polymer matrix with the solvent mixture. Stronger for modified hybrid ionogels.The glass transition temperatures arameter , no depeT5%) and 10% (T10%) of weight loss represent the onset temperature of the material decomposing [Thermal gravimetric analysis was used to investigate the thermal stabilities of synthesized ionogels. omposing .maxF) of synthesized ionogel poly(TATT + PETMP) with 70 wt.% of IL + PC is equal to 247 g. Modification with methacryl-POSS caused an increase in the values of this parameter. The addition of methacryl-POSS in the amount of up to 1.5–5.0 wt.% causes an increase in the parameter with an increase in modifier concentration, reaching the maximum value of ca. 330 g—there is no statistically significant differences in puncture strength for ionogels containing 1.5–5 wt.% of methacryl-POSS (results of the post hoc test are presented in maxε).Flexible and quite mechanically strong materials as for gels were obtained, i.e., ionogels can be twisted or rolled-up without damaging it. Their mechanical properties were investigated as puncture resistance. As can be seen in −1 at methacryl-POSS content equal 1–3 wt.%—there is no statistically significant differences in ionic conductivity for ionogels containing 1–3 wt.% of methacryl-POSS  ionic conductivity. This observation can be explained by the promotion of ion-pair dissociations in IL clusters caused by methacryl-POSS [Next to mechanical properties, ionic conductivity is an important parameter that decides the utility of ionogels as GPE in EC. In ryl-POSS .Considering mechanical properties  and ioniThe Nyquist plot of the AC/AC capacitor a equival−1 to 4 A∙g−1 causes linear decreasing of capacitance from ca. 144 F∙g−1 to ca. 78 F∙g−1 and from ca. 147 F∙g−1 to ca. 88 F∙g−1 for EC with ionogel containing 0 wt.% and 1.5 wt.% of methacryl-POSS. Similar behavior can be observed in the dependence of capacitance vs. scan rate (−1 to ca. 3400 W∙kg−1 (0 wt.% of methacryl-POSS) and from ca. 460 W∙kg−1 to ca. 3600 W∙kg−1 (1.5 wt.% of methacryl-POSS), causes only a slight decrease of energy density from ca. 67 W∙kg−1 to ca. 40 W∙kg−1 and from ca. 72 W∙kg−1 to ca. 42 W∙kg−1 for 0 wt.% and 1.5 wt.% of methacryl-POSS, respectively. Comparing the results obtained for capacitors with both ionogels, we can clearly see that ECs present similar capacitance/energy at a low value of current density (I), scan rate (v), or power (P). Thus, with increasing of these parameters  we can observe the greater difference in obtained values of capacitance and energy, with better results obtained for EC with ionogel containing 1.5 wt.% of methacryl-POSS. Furthermore, we should consider that both ionogels have the same thickness, and the ionogel containing 1.5 wt.% of methacryl-POSS shows better mechanical properties by ca. 30% at the same time. If we would like to apply a modified ionogel with similar mechanical properties as unmodified, we can reduce the thickness by about 30% and likewise reduce the resistance of the modified ionogel by about 30% (1.5 Ω). This will allow us to further improve the electrochemical properties of the EC.The CV of the AC/AC capacitor b obtainecan rate d, where can rate c presentAll these results show that modification of ionogels by 8M-POSS modifier and its application as GPE in electrochemical capacitor allows obtaining EC with very good electrochemical properties.2 and PC. Solvents mixture accelerates the thiol-ene polymerization of the TATT + PETMP system which was attributed to the polarity of the solvent which lowers the barrier for activation of hydrogen transfer in thiol-ene polymerization. On the other hand, methacryl-POSS decreases thiol-ene polymerization rate because of the low homopolymerization rate of methacryl-POSS.The hybrid ionogels modified with methacryl-POSS were successfully prepared by thiol-ene photopolymerization in a mixture of the imidazolium IL EMImNTfThe main disadvantage of the unmodified ionogel is leakage which makes the ionogel impossible to use as a gel polymer electrolyte. However, the modification with methacryl-POSS made it possible to obtain ionogels with no leakage. In addition, methacryl-POSS contributed to the improvement of the mechanical properties of hybrid ionogels. An increase in puncture strength (by approx. 30%) was observed, as well as a slight increase in the glass transition temperature. This is related to the steric hindrance of the POSS cages as well as the increase of cross-linking density of the polymer matrix. Additionally, the size of the interconnected polymer microspheres formed in dispersion polymerization decreases with the increase of methacryl-POSS content. The reason is poorer solubility of the modified polymer in the polymerizing medium at a similar conversion, so nucleation begins at lower conversions. Modification of ionogels with methacryl-POSS also causes an increase in ionic conductivity, by the promotion of ion-pair dissociations in IL clusters caused by methacryl-POSS. Moreover, application of hybrid ionogel as an GPE in an electrochemical capacitor allows obtaining EC with very good electrochemical properties.