Bacterial bone infections after revision surgeries and diseases, like osteomyelitis, are still a challenge with regard to surgical treatments. Local bone infections were treated with antibiotics directly or by controlled drug-releasing scaffolds, like polymethylmethacrylate (PMMA) spheres, which have to be removed at a later stage, but there is a risk of a bacterial contamination during the removement. Therefore, biomaterials loaded with antibiotics for controlled release could be the method of choice: The biomaterials degrade during the drug release, therefore, there is no need for a second surgery to remove the drug eluting agent. Even non-resorbable bone materials are available ) or resorbable bone graft materials ) that will be replaced by newly formed bone. Composite materials with organic additives  supports the handling during surgery and enhances the drug loading capacity, as well as the drug releasing time. The purpose of this study was to investigate the loading capacity and the release rate of Vancomycin and Gentamicin on TCP and HA granules in the shape of a degradable scaffold compared to composite materials from TCP mixed with porcine collagen. Its antibacterial efficacy to a more elementary drug with eluting in aqueous solution was examined. The loading capacity of the biomaterials was measured and compared according to the Minimum Inhibition Concentration (MIC) and the Minimum Biofilm Eradication Concentration (MBEC) of a bacterial biofilm after 24 h aging. Antibiotic elution and concentration of gentamycin and vancomycin, as well as inhibition zones, were measured by using the Quantitative Microparticle Systems (QMS) immunoassays. The antibiotic concentration was determined by the automated Beckman Coulter (BC) chemistry device. For examination of the antibacterial activity, inhibition zone diameters were measured. Generally, the antibiotic release is more pronounced during the first couple of days than later. Both TCP granules and HA granules experienced a significantly decline of antibiotics release during the first three days. After the fourth day and beyond, the antibiotic release was below the detection threshold. The antibiotic release of the composite material TCP and porcine collagen declined less drastically and was still in the frame of the specification during the first nine days. There was no significant evidence of interaction effect between antibiotic and material, i.e., the fitted lines for Gentamycin and Vancomycin are almost parallel. During this first in vitro study, β-TCP-Collagen composites shows a significantly higher loading capacity and a steadily release of the antibiotics Gentamycin and Vancomycin, compared to the also used TCP and HA Granules. The treatment of osteomyelitis is still facing challenging problems in orthopedic surgery. By reaching a chronic state of osteomyelitis, the local effectiveness of systemic anti-infectives is reduced due to a diminished local vascularity . AdditioDifferent methods and various grafting materials are used for the purpose of bone reconstruction . Among tCollagen-ceramic composites as synthetic bone regeneration materials have been developed for non-weight-bearing applications and the addition of collagen confers a number of advantages. These are built upon the material properties of the composite having the hemostatic function that allows early wound stabilization, attracts fibroblasts, and facilitates nutrient transport ,7. As thThe local application of antibiotics, besides surgical debridement, represents a viable alternative treatment to handle osteomyelitis by reaching higher local antibiotic concentrations and minimizing systemic side effects . Gentamicin and Vancomycin are common antibiotics in use for the treatment of bacterial infections. Because of the increasing resistance vancomycin is frequently used because of its antimicrobial activity against Gram-positive bacteria together with gentamicin against gram-negative bacteria as main targets . Both suFor the successful treatment of the infection, particularly when glycosides were used , is the Van de Belt described the release kinetics from polymethylmethacrylate bone cements as a combination of surface properties and porosity, which shows a biphasic process (burst-sustained release) in a study . After aThe intention of this in vitro study was a comparing examination of different antibiotics in combination with commercially available bone graft substitutes. The antibiotics, as well as the biomaterials, do have different structures and therefore the combination of both are different in dependence with the material , the structure  and pharmaceutical agent. The preparation and the uptake of the antibiotic formulations under clinical conditions were simulated by dripping the solutions onto the biomaterials and measuring the loading capacity. The loading capacity was measured in order to understand which amount of antibiotics can be combined for clinical applications. For drug release experiments, the biomaterials were eluted in aqueous solution, and the concentration of the pharmaceutical active substance was measured during elution time in the surrounding liquid. Two experimental setups were implemented in order to measure the release behavior of different Gentamycin and Vancomycin-doped ceramic which was based on bone substitute . The conventional test setup is dipping the loaded biomaterials in an aqueous solution and measuring the drug release as a function of time by measuring the concentration in the solution. This is a dilution method to understand the interaction between antibiotic and biomaterial. Another test method is dripping the above described solution onto an agar plate with bacterial layer by measuring the inhibition zone around the laden biomaterial as a function of time. So, the antibacterial activity and function of the antibiotics, first laden on the biomaterial and then released in the aqueous solution, is monitored.®) and β-tricalcium phosphate granulate with a porosity of 65% (Cerasorb® M) were both used with a granulate size of 1000–2000 µm. β-tricalcium phosphate composite materials consisted of 85 vol % porcine collagen and 15 vol % β-tricalcium phosphate (Cerasorb® Flexible Foam) with a density of (0.4 ± 0.1) g/cm3 was used in this work. Depending on its density, the materials had different mechanical characteristics. All materials were provided by Curasan AG, Kleinostheim, Germany.Open cellular hydroxyapatite (HA) granulate with a porosity of 75% (OsboneThe scaffolds were examined by Scanning Electron Microscopy (SEM) in order to determine the surface and porosity characteristics of the biomaterials. The samples were placed on a sample holder using conductive carbon tape followed by sputtering with graphite and gold. The materials were examined using an AMRAY Scanning Electron Microscope Type 1810P . Photos were filed using different magnifications depending on the surface characteristic of the material. The pore sizes were measured using the scale bars. The density and porosity of the materials was determined by measuring the bulk density of the scaffolds.The QMS Gentamicin and Vancomycin assays are homogeneous particle-enhanced turbidimetric immunoassays. The assays are based on competition between drug in the sample and drug coated onto a microparticle for antibody binding sites of the antibody reagents. The antibiotic-coated microparticle reagent is rapidly agglutinated in the presence of the anti-antibody reagent and in the absence of any competing drug in the sample. The rate of absorbance change is measured photometrically. When a sample containing the antibiotic is added, the agglutination reaction is partially inhibited, slowing down the rate of absorbance change. A concentration-dependent classic agglutination inhibition curve can be obtained with maximum rate of agglutination at the lowest antibiotic concentration and the lowest agglutination rate at the highest antibiotic concentration. The antibiotic concentration was determined by the automated Beckman Coulter AU480-clinical chemistry device .® Foam pellets were cut from the scaffolds. The granules of biomaterials were used in the original structure. Therefore, five samples of every sort of the respective bone graft substitutes Cerasorb® M granules , Cerasorb® Foam , and Osbone® granulate  were soaked with 4 mL of the antibiotic solutions (Gentamicin-Ratiopharm 80 SF (4% solution), ratiopharm GmbH, Ulm, Germany, 40 mg/mL); and Vancomycin 50 mg/mL (5% solution), Hikma Pharma GmbH, Gräfelfing, Germany) for 1 min at room temperature in a petri dish under simulated clinical conditions.For each experiment there was 100 mg biomaterial used. CerasorbAntibiotic in wt %), the following Equation (1) was used:Beckman Coulterc—Measurement of AU480 in mg/L in 5 mL PBS substrate, and Biomaterialm—Initial weight of the biomaterial sample.The total antibiotic uptake of the pellets was carried out by pestling the pellets and loading with antibiotics. Subsequently, 5 mL PBS was also added and followed by incubation for 24 h at 37 °C. The supernatant was measured with the Beckman Coulter AU480 device as described in From these data, the minimum biomaterial amount with uptaken antibiotics to reach the MBEC, according to Marques et al., in mg was calculated .TM, Rockford, IL, USA). The detection limit for the QMS was 2.5 µg/mL for Vancomycin and 0.5 µg/mL for Gentamycin. Additionally, disk diffusion was used to measure the effect of the antibiotic release on microorganisms. The release kinetics of the antibiotics Gentamicin and Vancomycin with regard to the bone graft substitutes was determined. The released antibiotic concentrations were measured using the Quantitative Microparticle Systems (QMS) Gentamicin and Vancomycin immunoassays , pH 7.4, at 37 °C for 24 h. The release kinetics of each of the two antibiotics was measured in multiple test runs (n = 5) on 10 consecutive days. Each day, the supernatant was taken from the solution. One hundred microliters was used for disk diffusion, and the rest of the solution was preserved at −80 °C for subsequent determination of the antibiotic concentration. The pellets were washed with PBS and then, again, 10 mL PBS were added, followed by subsequent incubation.Outgoing from the release percentage after 24 h the minimum amount of Antibiotic-loaded biomaterial was calculated to reach MBEC after 24 h.For disk diffusion, a bacterial suspension of the test organisms Bacillus subtilis ATCC 6633 and Staphylococcus aureus ATCC 25923 was adjusted using a McFarland standard of 0.5 in 0.9% sterile saline and streaked over the whole surface of a Columbia blood agar plate with a sterile swab. One hundred microliters of the supernatant of the incubated bone graft substitutes was transferred in the middle of the plate. After overnight incubation at 35 ± 2 °C, the zone diameters were measured.® Analytic Software  and Solutions to fit nonlinear fixed models. Multiple measurements on each specimen were taken into account by including a random specimen effect in the model.In order to describe the antibiotics release as a function of time, two evaluation methods were used. Outgoing from the Antibiotic concentrations in the supernatant measured by the Beckman Coulter Device  in mg/L. 100%-release of the drug was supposed when no change of the amount was detected. The release was then plotted in cumulative percent release. In a second method, piecewise linear functions to the antibiotics release were fitted, i.e., In order to describe the dependence of the inhibition zones for staphylococcus aureus and bacillus subtilis on antibiotics release, a linear mixed model (SAS PROC MIXED) was used, taking multiple measurements on each specimen into account by including a random specimen effect in the model. The dependent variable was the area of the inhibition zone, which was calculated from the diameter, assuming a circular shape. Independent variables were antibiotics’ release, material, and type of antibiotic. Here, observations with antibiotics release below the detection threshold were excluded from the model, as these observations neither had a detectable inhibition zone.Antibiotic uptake was compared, using the Kruskal–Wallis test and Wilcoxon–Mann–Whitney test, and exact p-values were determined. All analyses were performed using SAS 9.4.® M showed a rough surface with statistical distributed pores from 0.2–500 µm. The inner surface is enhanced by this interconnecting micro-, meso-, and macroporosity.® shows a bone-like structure. Pores are replaced by open cellular structure formed by bars of 10–50 µm thickness. These bars are formed by the primary HA particles forming a bone-like nearly 80% porous structure with a medium inner surface.Osbone® Foam shows a fibrous structure of collagen fibers. The ceramic is completely embedded in this structure and is not visible by this examination method. The material is open-structured, highly porous, and has a maximized inner surface. Due to the fibrous structure of the organic phase, the material has a high internal liquid uptake characteristic.Cerasorb® Granules having the lowest uptake, Osbone® Granules an intermediate intake, and Cerasorb® Foam having the highest intake . One measuring value with Cerasorb® Foam was an outlier value and, therefore, discarded from the analysis. When comparing antibiotics, no difference in uptake could be demonstrated for Cerasorb® Foam and Osbone® Granules. However, for Cerasorb® Granulate the Vancomycin uptake was clearly higher than the Gentamycin uptake , steeper slopes in the first part , and more gentle slopes in the second part . There was no significant interaction effect of antibiotic and material, i.e., the fitted lines for Gentamycin and Vancomycin are almost parallel . However, there may exist a threshold, especially in Cerasorb® Form compared to the granules . Again, there may exist a threshold, especially in Cerasorb® Foam than for the granulates  pellets, in comparison to polymethylmethacrylate (PMMA) beads ,17. Pure® M Granules and the Collagen composite Cerasorb® Foam, were loaded with different concentrations of Vancomycin solution from the same origin like in this work. As distinguished from this work the biomaterials were loaded with a dripping technique under normal conditions as well as in vacuum to obtain a more and a complete soaking of all pores and interstices of the biomaterials. The incubation was then carried out in a drying oven at 50 °C for 96 h. Due to this dripping and drying technique a higher load of pharmaceutically active substance could be reached. The release kinetic did not show the strong initial bursts like in this actual work and had a much longer release kinetic: even after 14 days incubation period nearly 50% of the antibiotic remained in the biomaterial caused by a stronger binding of the substance on the inner surface of the biomaterials. With this way the antibiotic concentrations around the materials are at a nearly constant level over long periods and could be helpful for clinical applications. The remaining active substance will be released during the biodegradation/resorption of the biomaterials. Additional experiments of controlled drug release with Vancomycin from these biomaterials under lower pH conditions, simulating the treatment of infected areas showed an irregular release kinetic [In a previous work, different biomaterials, amongst others also Cerasorb kinetic . In this2/g vs. 80 m2/g). This opens a huge potential for physisorption of the antibiotics. The big amount of functional side chains in collagen with polar COOH- and NH2- groups should also provide significant ionic and electronic interdependencies. The question of why Vancomycin, compared to Gentamycin, demonstrates a stronger bonding to the provided matrix can obviously be answered by the higher molecular weight of Vancomycin . The good adsorption to the composite matrix can be explained by multiple effects: On the one hand, the surface of collagen is much bigger than it is in Calcium phosphates , while a larger quantity is needed to reach the MBEC (Minimum Biofilm Eradication Concentration). In all cases, the drug-released kinetic started with an initial burst reaching the MIC in the surrounding liquid in the first 24 h. A slower release of the pharmaceutical agents took place in a second step. The drug release of the granulated material was faster compared to the collagen composite material. In all cases, the pharmaceutical agent was practically released to 100%. The antibacterial activity of the antibiotics were measured after the loading and release from the biomaterials, which was still showing an inhibitory effect of Staph. Aureus and Bacillus subtilis. β-TCP-Collagen Composites showed in this first in vitro study a significantly stronger and longer release of Gentamycin and Vancomycin compared to β-TCP- and hydroxyapatite granules. For clinical use, a more sustainable therapy against sensitive exciters is to be expected. The osteoconductivity of the composite may offer an additional positive effect in the treatment of bone infections.


Human hepatocellular carcinoma (HCC) lacks effective curative therapy and there is an urgent need to develop a novel molecular-targeted therapy for HCC. Selective tyrosine kinase inhibitors have shown promise in treating cancers including HCC. Tyrosine kinases c-Met and Trks are potential therapeutic targets of HCC and strategies to interrupt c-Met and Trks cross-signaling may result in increased effects on HCC inhibition.The effects of Indo5 on c-Met and Trks activity were determined with in vitro kinase activity assay, cell-based signaling pathway activation, and kinases-driven cell transformation. The in vivo anti-tumor activity was determined with xenograft mice and liver orthotopic mice models. The co-expression of c-Met and TrkB in 180 pairs of HCC and adjacent normal tissues were detected using immunohistochemical staining.Indo5, a novel lead compound displayed biochemical potency against both c-Met and Trks with selectivity over 13 human kinases. Indo5 abrogated HGF-induced c-Met signaling activation and BDNF/NGF-induced Trks signal activation, c-Met or TrkB-mediated cell transformation and migration. Furthermore, Indo5 significantly decreased the growth of HCC cells in xenograft mice and improved the survival of mice with liver orthotopic tumors. In addition, co-expression of c-Met and TrkB in HCC patients was a predictor of poor prognosis, and combined inhibition of c-Met and TrkB exerted a synergistic suppressive effect on HCC.These findings indicate that Indo5 is associated with marked suppression of c-Met and Trks co-expressing HCC, supporting its clinical development as an antitumor treatment for HCC patients with co-active c-Met and Trks signaling.The online version of this article (10.1186/s13046-019-1104-4) contains supplementary material, which is available to authorized users. Selective tyrosine kinase inhibitors have shown promise in treating cancers driven by activated tyrosine kinases such as EGF receptor (EGFR) in non-small cell lung cancer (NSCLC), Bcr-Abl in chronic myelogenous leukemia (CML), and c-Kit in gastrointestinal stromal tumors (GIST) . SorafenOngoing efforts to study hepatocarcinogenesis have identified an important role of c-Met signaling in the promotion of tumor growth, angiogenesis, and metastasis including HCC. c-Met transcription is increased in HCC tumors and overexpression c-Met receptor protein results in a poor prognosis . In addiAccumulated evidence have reported that aberrant c-Met activation can occur in various tumors through crosstalking with other receptors, including EGFR, chemokine receptor 4 (CXCR4), VEGFR-2 and FGFR . MoreoveThe tropomyosin receptor kinase (Trk) family belongs to receptor tyrosine kinases and is composed of three homologous receptor tyrosine kinases: TrkA, TrkB, and TrkC, which specifically bind neurotrophins nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin 4 (NT4) and neurotrophin 3 (NT3), respectively . NeurotrRecently, several lines of evidence have revealed a connection between c-Met and Trk receptors in promoting tumor progression in addition to their respective roles in the occurrence and development of tumors. Activated c-Met enhances TrkA signaling and the bioactivity of NGF , and TrkHere, we described Indo5 as a potent and selective c-Met and Trk inhibitor. Importantly, Indo5 exhibited the antitumor effects on HCC with in vitro models, in vivo xenograft, and orthotopic models. These findings supported its clinical development as an antitumor treatment for HCC patients with co-active c-Met and Trks signaling.The human hepatocellular carcinoma cell lines HepG2, HCC LM3, MHCC97-L, MHCC97-H, BEL-7402, and SMMC-7721, human liver cell line L02, human neuroblastoma cell line SK-N-SH, human lung carcinoma cell lines A549 and NCI-H460, human fetal lung cell MRC-5, primary mice hepatocytes, and Madin-Darby Canine Kidney (MDCK) cell were all cultured in DMEM-H (Gibco) supplemented with 2 mM glutamine, 100 units/ml penicillin, 100 mg/ml streptomycin and 10% FBS. Murine pre-B cells (Ba/F3) were grown in RPMI 1640 containing 10% FBS and 10% WEHI-3 cell-conditioned medium as a source of murine IL-3. The compound 3-(1H-benzimidazol − 2-methylene)-5- (2-methylphenylaminosulfo)- 2-indolone (Indo5) was synthesized as previously described (US Patent No: 9642839B2). For oral gavage, Indo5 was dissolved in 0.5% CMC-Na.3 cells were seeded in a 96-multiwell plate . After 4 h, different concentrations of inhibitors were added to the wells. The plate was immediately put into the Operetta and incubated for 48 h at 37 °C and 5% CO2. Digital phase contrast images were sequentially acquired every 3 h using a 10 × objective with 6 fields per well. Image analysis was performed using the Harmony application to provide the algorithm.All cell growth assays were performed using the High Content Screening System (PerkinElmer). Briefly, 2 × 10Female BALB/c athymic nude mice aged 5 to 6 weeks were purchased from Beijing Vital River Laboratory Animal Technology  and allowed to acclimate for 1 week. Animal care was in accordance with the institutional animal care guidelines. All animal experiments were approved by the Animal Ethics Committee of the Academy of Military Medical Science.6 HepG2 cells, 4 × 106 MHCC 97H cells, 4 × 106MHCC 97 L cells, 2 × 106 A549 cells, or 5 × 106 SMMC-7721 cells in a volume of 200 μl PBS were inoculated subcutaneously in the right flank of each mouse. Tumor growth was measured using vernier calipers, and tumor volume (mm3) was calculated as (L × W × H)/2. When the mean tumor volume reached 200 mm3, the mice were randomized into four groups and treatment was initiated. Indo5 in DMSO was injected intraperitoneally into the mice once daily at the indicated doses. Tumor volume were recorded every 3 days until the study was terminated. The mice were sacrificed after 21 days of treatment. Solid tumors were excised and weighed, and the inhibition rate was calculated as (WeightIndo5 treatment group-Weightcontrol group)/Weightcontrol group.First, 2 × 103, the mice were sacrificed, and the tumors were isolated and cut into 1 mm3 pieces. The mice were anesthetized by an intraperitoneal injection of 1% sodium pentobarbital solution. A 1-cm longitudinal skin incision was made on the right upper axillary region of the abdomen of the mouse, the peritoneum was opened and the liver was exposed. The liver was punctured by a needle, and a 1 mm3 tumor piece was implanted. The liver was put back into the abdominal cavity gently, and the surgical opening was closed using a 5–0 surgical suture.For the orthotopic xenograft mouse model, MHCC97H cells were inoculated subcutaneously in the right flank of each SCID mouse. When the tumor volumes reached approximately 1 cmp-value of 0.05 or less was considered statistically significant.Data were expressed as the mean ± S.D. All statistical analyses were performed using the GraphPad Prism software. Statistical significance was calculated using a two-way ANOVA. A 3-(1H-benzimidazol-2-methylene)-5-(2-methylphenylaminosulfo)-2-indolone (Indo5) . Analysis of the time course of Indo 5 on the c-Met phosphorylation suggested that the inhibitory effect of Indo 5 was lasts from 2 h to 6 h . No significant effect of Indo5 was observed on other 13 tyrosine kinases. These data suggested that Indo5 may be a potent inhibitor for Trk kinases. To confirm, we investigated the effect of Indo5 on the Trk signaling pathway in SK-N-SH cells. The data suggested that Indo5 pre-treatment inhibited NGF-induced signaling in a dose-dependent manner, and TrkA signaling was almost completely inhibited at a concentration of 1 μM Indo5 , the IC50 value of Indo5 ranged from 1.114 ± 0.018 μM to 1.873 ± 0.012 μM. In cells expressing TrkB alone or both low (c-MetloTrkBhi and c-MetloTrkBlo), the IC50 value is much higher, ranging from 2.982 ± 0.019 μM to 9.185 ± 0.035 μM  even in L02 cells the expression levels of c-Met and TrkB were high. These results indicated that the inhibition effect of Indo5 on HCC growth was closely correlated with the expression levels of c-Met and Trk.We further investigated the effect of Indo5 on the growth of these cell lines. Indo5 significantly inhibited cell growth in a dose-dependent manner in c-Met and TrkB coexpressing cell lines . Thus, we showed that Indo5, at well-tolerated doses, suppressed HepG2 xenograft growth in nude mice. Similar results were obtained in MHCC97H, MHCC97L, and A549 xenograft mice  staining with specific c-Met and TrkB antibodies. Figure high TrkB low (n = 83), c-Met high TrkB high (n = 36), c-Met low TrkB low (n = 57), and c-Met low TrkB high (n = 4). Since the patient number in the last group was too small for statistical analysis, we analyzed the overall survival in the other three groups. As shown in Fig. high TrkB low group exhibited poor overall survival (OS) compared to patients in c-Met low TrkB low , and the group c-Met high TrkB highhad shorter OS times than did the group c-Met high TrkB low . These data indicate that the co-upregulation of c-Met and TrkB may be a potential factor for predicting the prognosis of HCC.To further evaluate the significant contribution of c-Met and TrkB coexpression in the prognosis of patients with HCC, 180 patients with HCC were divided into four groups according to the c-Met and TrkB expression levels in their tumors: c-Met igh n = 3, c-Met lIn this report, we identify Indo5, a novel indolinone derivative, as a tyrosine kinase inhibitor that targets c-Met and Trks in vitro and in vivo. Indo5 potently inhibited proliferation and survival of Tpr-Met and TrkB transfected Ba/F3 cells. Moreover, HCC cell lines with high c-Met and Trks coexpression were more sensitive to Indo5 and inhibition of c-Met and TrkB significantly reduced the antitumor activity of Indo5. These results suggested that the antitumor activity of Indo5 in HCC is mediated by suppression of oncogenic signaling of c-Met and TrkB. Of special interest was the observation that Indo5 treatment improved the survival of mice with liver orthotopic model generated by MHCC97H cells, which expresses high levels of c-Met and TrkB, with Indo5 showing similar or superior activity to Sorafenib. In addition, we showed that co-expression of c-Met and TrkB in HCC patients was a predictor of poor prognosis, and combined inhibition of c-Met and TrkB exerted a synergistic suppressive effect on HCC. These findings indicate that Indo5 is associated with marked suppression of c-Met and Trks co-expressing HCC, supporting its clinical development as an antitumor treatment for HCC patients with co-active c-Met and Trks signaling.50 values of 14.37 nM, 28 nM, and 25 nM, respectively. Furthermore, we noticed that while Indo5 displayed similar IC50s against the kinase activity of TrkA and TrkB in vitro (Table S1), it seemed that TrkB was more sensitive to TrkA in cells. This differential activity of Indo 5 on the signaling mediated by TrkA and TrkB might due to a different response to NGF and BDNF of SK-N-SH and HepG2 cells. No significant effect of Indo5 was observed on other tyrosine kinases including EGFR, VEGFR, FGFR1 and c-Met family member, Ron. Although the specificity of Indo5 kinase inhibition still requires further studies, our findings provide evidence that Indo5 exerts a potent cytotoxicity toward subsets of HCC cells characterized by high c-Met and Trk co-expression levels. In the development of tyrosine kinase inhibitors, a balance between toxicity and efficacy must be achieved. In many cases, the ideal anticancer effect can be achieved by inhibiting multiple kinases, which in turn increases its toxicity. In our experiment, while sorafenib treatment induced up to 10% body weight loss, and Indo5 treatment was not associated with loss of body weight even at the highest dose as single agent (48 mg/kg), indicating Indo5 was well tolerated in mice.Indo5(3-(1H-benzimidazol-2-methylene)-5-(2-methylphenylaminosulfo)-2-indolone) contains the indolone core structure, which have been identified as a versatile scaffold for the development of protein kinase inhibitors . By alteThe role of HGF/c-Met and BDNF/TrkB signaling is reported in several cancers including HCC. Potential crosstalk between these signaling pathways is described in neuroblastoma , salivarCombination targeting c-Met and other receptors exhibited a better anti-tumor effect than single agent administration. We showed that combined inhibition of c-Met and TrkB with siRNA or specific inhibitors exerted a synergistic suppressive effect on growth and migration of HCC cells, indicating that combined targeting of c-Met and Trk kinases is a useful therapeutic strategy. We demonstrate that co-expression of c-Met and TrkB is present in about 20% HCC patients and association with poor prognosis, indicating that co-expression of c-Met and TrkB may be a biomarker for HCC personalized therapy. Base on our above studies, we speculate that in a subclass of HCC patients with co-upexpression of c-Met and TrkB might benefit from targeted therapy with c-Met and Trk inhibitors. In addition to Indo5, several compounds have been shown to suppress c-Met and Trks such as K252a , CabozanWe have identified Indo5 as a novel inhibitor targeting c-Met and Trks. Our results show that Indo5 suppresses the growth of HCC cells with high levels of c-Met and Trk, providing a potentially new and practical pharmacological approach for treating HCC patients with co-active c-Met and Trks signaling.Additional file 1:A selective c-Met and Trks inhibitor Indo5 suppresses hepatocellular carcinoma growth. (DOC 20 kb)


Based on the fresh-state tests, including 3D printing as such, an optimum composition was identified, and its compressive and flexural strengths were characterized. The printable foam concrete showed low thermal conductivity and relatively high compressive strengths of above 10 MPa; therefore, it fulfilled the requirements for building materials used for load-bearing wall elements in multi-story houses. Thus, it is suitable for 3D-printing applications, while fulfilling both load-carrying and insulating functions. Three-dimensional (3D) printing with foam concrete, which is known for its distinct physical and mechanical properties, has not yet been purposefully investigated. The article at hand presents a methodological approach for the mixture design of 3D-printable foam concretes and a systematic investigation of the potential application of this type of material in digital construction. Three different foam concrete compositions with water-to-binder ratios between 0.33–0.36 and densities of 1100 to 1580 kg/m Foam concrete of density lower than 400 kg/m3 is used primarily as a filling or insulating material [Foam concrete (FC) is a lightweight cementitious material with a cellular structure produced by incorporating air voids into mortar or cement paste. It can be designed to have a density in the range of 200 to 1900 kg/mmaterial . 2 footprint and very limited recyclability. In contrast to such materials, foam concrete is made of mineral constituents with only minor contents of chemical admixtures [Groups working with foresight on digital fabrication have identified the future need for sustainable construction materials that are economically efficient and environmentally friendly . It is emixtures . AdditioThe Concrete on-site 3D-Prining (CONPrint3D) concept developed at the Technische Universität Dresden facilitates implementation of the advantages of the additive technology in the construction industry . In contThe literature contains an example that describes an automated application of foam concrete to vertical surfaces by means of an extrusion-based technique . The aut3 and compressive strength in the range of 1.5 to 9 MPa, and which in addition maintains its dimensional stability after extrusion. The water-to-cement ratio (w/c) was set to 0.3 in all the mixtures. Neither fillers nor aggregates were used. Preformed foam was prepared with a protein-based foaming agent. The research provides a wide range of results related to the influences of curing conditions on tensile and compressive strengths. However, the experimental procedure described did not represent typical 3D-printing procedures by robotic print heads. The material was rather filled into steel formwork and pushed down manually from the formwork in the early stage of hydration. The deposition technique used by Faliano et al. imitated automated extrusion and provided first filling of the material behavior in terms of form stability and green strength development. Faliano et al. ,15 descr3. Both structural and non-structural applications for the extrudable elements made of foam concrete were denominated as effective and environmentally friendly. One of the suggested applications was to form multilayer insulating panels in situ.There is no standard way of measuring the properties of buildability. Generally, buildability is evaluated by printing a certain number of layers at a specific rate ,17,18,19In general, concrete that is suitable for digital construction must be well extrudable and demonstrate adequate buildability. Furthermore, printed layers need to have good interlayer bonds ,16,20,21Another important feature of a printable material is its buildability, which is comprised of the shape stability of the printed layers under their self-weight and the ability to hold further layers with minimum deformation . In otheIn studies related to 3D printing with normal-weight concrete, quick setting is usually achieved by using accelerating admixtures or by choosing cements with shorter setting times, i.e., rapid-hardening sulfoaluminate or calcium aluminate cements ,25. The Another important aspect of printed elements is their interlayer bonding. It strongly influences the mechanical properties, durability, and serviceability of 3D printed structures; see e.g., ,31,32. TGenerally, there are two mechanisms to introduce large volumes of air voids into the mixture: (1) the use of gas-forming chemicals such as aluminum powder, and (2) the use of foaming agents. Addition of the gas-forming agents results in bubble formation through chemical reactions with alkaline hydration products, e.g., calcium hydroxide . This meIn the alternative approach, foam concrete can be produced either by adding the foaming agent to the cement paste, followed by intense mixing, which is called the mixed foaming method, or by intermixing a separately produced foam into cement paste, which is known as the prefoaming method ,4. In coThe mixed foaming method is widely used in the construction industry to produce foam concrete. However, this method is limited according to the use of the synthetic foaming agents and is highly dependent on the mixing device used. In contrast, the prefoaming method enables the defining of the material density by the exact addition of the required amount of the foam to the base mixture. Since the ratio of foam to base material could be greater than 1:1, foam becomes a major influencing factor . The sta3. Finally, the insulating properties of the printable foam concrete were compared to those of normal-weight printable concrete . The scheme of the mixture design approach as developed in the framework of the research project CONPrint3D-Ultralight is presented in Firstly, constraints such as the range of the water-to cement ratio (w/c) and cement content must be set according to the intended application. Based on information from the literature, suitable proportions and materials can be identified. Production and characterization of the foam follow. The aim of this step is to obtain sufficiently stable foam that is capable of withstanding the intermixing process. In parallel, the water demand and binder composition of the cement-based matrix, including the superplasticizer (SP) dosage, are determined by iterative testing. Workability was assessed by measuring the spread flow diameter values in accordance with the European standard DIN EN 1015-3:1998, and thus using the so-called Hägermann cone and by applying 15 strokes . In the The third step focuses on verifying the fresh foam concrete’s rheological properties, which have to comply with the 3D-printing process requirements of printability, extrudability, and buildability ,40,41,42The last step specifies testing the hardened state properties of foam concrete, such as its compressive and flexural strengths, thermal conductivity, and/or durability. At this stage, the water-to-binder ratio (w/b) may be reduced; alternatively, reinforcement in the form of dispersed nanofibers or microfibers may be introduced ,3,43. ThIt is essential to specify a suitable water content in the foam concrete. There is no standard procedure, especially when the requirements of printability, pumpability, and buildability must be fulfilled. In the present work, the water demand of the cement-based matrix was determined by Okamura’s and Ozawa’s method . The com2 and Al2O3. The implementation of fly ash into concrete composition on the one hand enabled decreasing the water demand of dry constituents while still achieving the targeted rheological behavior; on the other hand, it improved the sustainability of the mixtures. A polycarboxylate ether (PCE)-based SP  was used in the cement-based matrix to adjust the workability at reduced water content. The water content in the SP was 77% by mass. The density of the SP equaled 1050 kg/m3. A protein-based foaming agent  was utilized for production of the foam. A type II Portland composite cement CEM II/A-M (S-LL) 52.5 R  was used. Hard coal fly ash Steament H-4  was chosen as a secondary cementitious material. The chemical composition and measured particle size distribution are provided in In a preliminary stage, three liters of the cement-based matrix paste were produced to assess the water demand using a pan mixer . Foam concrete was produced using a conical, multi-rotor colloidal mixer . For each experiment, 30 L of foam concrete was prepared using the procedure according to Extrusion and deposition experiments were conducted using two devices: (a) a standalone progressive cavity pump (PCP1) DURAPACT DP 326S , and (b) the 3D concrete printing test device  equipped with PCP2; see Each printed wall was transferred to a climatic chamber at an age of 24 h and cured at a constant temperature of 20 °C, a relative humidity of 65%, and in the absence of wind for 27 days. This procedure on purpose does not accord to DIN EN 12390-2 , which p3 were saw-cut from the walls printed in the same manner as those for mechanical testing. The insulation properties of the optimum mix composition were measured with a Heat Transfer Analyzer ISOMET 2104 . This apparatus applies a dynamic measurement method that enables the reduction of the period of the thermal conductivity measurements to a mere 10 to 16 min. Specimens of dimensions of 70 × 70 × 20 mmScanning electron microscopy (SEM) was used to visualize the microstructure of the foam concrete. The SEM unit Quanta 250 FEG  was operated in the so-called “low-vacuum mode”, whereby the non-conductive specimens were imaged as obtained without sputter-coating. 4). This part of the sample preparation is in accordance with DIN EN 480-11:2005 [The pore structure of foam concrete consists of gel pores, capillary pores, and entrained and entrapped air voids . Gel and-11:2005 . An areaThe water demand idelines ,49, onlyThe water demand of self-compacting concrete following Okamura’s procedure is usually set at 80% to 90% of the experimentally determined g factor , but firFor the first foam concrete experiments, composition A-1, consisting of cement and fly ash, was chosen. The foam volume was calculated according to the method described in , which dThe spread diameters of all the mixtures comply satisfactorily with the requirement for printable concrete, which was shown to be between 120–140 mm . Figure A common challenge in foam concrete production is the instability of its pore system and the resulting deviation from the targeted density, which is hereafter termed density discrepancy. The risk of such discrepancy is increased in the case of printable foam concretes due to their relatively stiff cement-based matrix. Therefore, the mixture design approach of the study at hand incorporates a step for the revision of the concrete composition based on this discrepancy in the density achieved, as compared to the initial target density. 3 and 1180 kg/m3, respectively. Similarly, M-3 had a fresh density of 1430 kg/m3, which differed markedly from the target value of 1580 kg/m3. Interestingly, the density discrepancy of M-2 was almost negligible in comparison to that of the other mixtures. The results presented in [Mixture M-1 had an initial target density and a finally measured fresh mixture density of 1100 kg/mented in  indicate3 (M-3) had a weight loss upon drying of just about 9%, higher amounts of foam added gave rise to pronouncedly higher drying losses of approximately 18% (M-1 and M-2). The density of the foam concretes changes over time due to drying. The kinetics of this process and the resulting dry density depend on ambient temperature and humidity as well as on maturity, fresh density, and w/c of the concrete, and on the area of the surface exposed to evaporation. The drying weight loss varied with the density; see All the mixtures could be extruded and deposited during the first 20 min after production without any visible deformations upon deposition. During the preliminary investigation, M-1 layers were extruded and deposited manually; see The buildability of the M-2 and M-3 foam concretes was evaluated by printing a maximum number of layers until collapse of the wall specimens. The maximum heights and number of layers of printed walls with M-2 and M-3 equaled 15 cm with 16 layers and 10 cm with six layers, respectively. Note that the geometries of the nozzles varied between M-2 and M-3, and the aspect ratios (height to width) were 0.2 and 0.7, respectively. The edges of all the layers retained the rectangular default shape imposed by the nozzle, and no significant deficiencies were noted on the surfaces of the deposited layers. In the observation period of 24 h after printing, no visible cracks or other signs for early age degradation were found. Of the three compositions designed, M-2 revealed the lowest discrepancy from the target density; see also 3 and the printed concrete with a density of 2100 kg/m3 exhibited similar thermal conductivities of 1.70 W/mK and 1.68 W/mK, respectively. Contrarily, printable foam concrete M-2 with its closed-cell meso structure was found to have a thermal conductivity of 0.24 W/mK, which is approximately 85% lower than that of the reference concretes; see also A higher magnification of matrix regions discloses that the microstructure is fairly dense and does not have significant capillary porosity, which would be in the range of pore diameters of some few up to tens of micrometers; see The walls between the air voids constitute the load-carrying frame in the structure of the foam concrete. Interestingly, no cracks originated or terminated at the large-scale air bubbles that stem from the foam. This finding indicates that local tensile strains due to different drying shrinkage degrees were insignificant, which is in contrast to . 3. These concretes could be extruded and deposited layer-wise.Based on an established approach to the mixture design of self-compacting concrete, it was possible to manufacture 3D-printable foam concretes in the density rage of 970 to 1500 kg/mThe discrepancy between the designed target density and that achieved was in the range of 0.25% to 10%. Improvement in the stability of prefabricated foam, especially a reduction in the time-dependent drainage of water, should contribute to even more robust compositions in consecutive research steps.The water-to-powder ratio of 0.37 of cement-fly ash paste in a proportion of 60:40 by volume with the addition of 0.25% of the SP by weight yielded an eminently workable cement-based matrix which is appropriate for the intermixing of prefabricated foam with minimal brakeage of bubbles.3, exhibited a compressive strength of 10.4 MPa after 28 days. Although these values are the results of laboratory-scale test specimens, they clearly indicate that printable foam concrete can prospectively be used for structural applications, e.g., load-bearing walls in multi-story houses. Further aspects such as testing the mechanical properties of real-scale prototypes need to be studied before the technology can be recommended for large-scale industrial implementation.Foam concrete composition M-2, with a dry density of 980 kg/mFrom a broader perspective, walls with high thermal insulation (currently 0.24 W/mK) could be produced using additive technology directly on the site.The potential application of foam concrete in digital construction was reviewed, and the mixture design for this novel material was developed and verified. The major findings of this work are as follows: 3 with appropriate mechanical characteristics for structural applications.Future work should focus on rheological behavior by varying the foam contents and quantitative characterization of the possible anisotropy of the mechanical properties of printed foam concrete. Production techniques for continuous foam concrete production, including the mixed foaming method, should be considered as well. It is expected to achieve raw densities of printable foam concrete in the range of 800 to 900 kg/m


The aim of this study was to identify fibrosis-related serological surrogate outcome measures in patients with immunoglobulin G4-related disease (IgG4-RD).This was a clinical observational study of 72 patients with untreated IgG4-RD from four institutions in Japan. The serum concentrations of growth differentiation factor 15 (GDF-15), CCL2, hyaluronic acid (HA), amino-terminal propeptide of type III procollagen (PIIINP), and tissue inhibitor of metalloproteinases 1 (TIMP-1) were measured by enzyme-linked immunosorbent assays. The enhanced liver fibrosis (ELF) score was calculated from the TIMP-1, PIIINP, and HA values. We evaluated associations between the values of these biomarkers and laboratory data, the IgG4-RD responder index (IgG4-RD RI) score, and organ involvements.Compared with the 44 healthy controls, the patients with IgG4-RD showed significantly elevated serum concentrations of GDF-15, MCP-1, HA, PIIINP, and TIMP-1 and ELF scores. The patients’ serum concentrations of GDF-15, CCL2, HA, and TIMP-1 (but not PIIINP) were positively correlated with each other. Among them, serum GDF-15 most efficiently distinguished patients with IgG4-RD from healthy controls. Serum GDF-15 was not associated with the IgG4-RD RI score or the number of organ involvements but was independently associated with the presence of retroperitoneal fibrosis and with parotid gland involvement.We observed increased serological surrogate outcome measures of fibrosis in IgG4-RD. GDF-15 may precisely reflect the fibrotic degree in patients with IgG4-RD.The online version of this article (10.1186/s13075-018-1777-7) contains supplementary material, which is available to authorized users. Immunoglobulin G4-related disease (IgG4-RD) is a fibro-inflammatory condition generally characterized by tumefactive lesions, a dense lymphoplasmacytic infiltrate rich in IgG4-positive plasma cells, storiform fibrosis, and  elevated serum IgG4 concentrations . IgG4-RDFibrosis and the infiltration of IgG4-positive plasma cells are predominant histopathological features of IgG4-RD. Although histopathological findings are the key to the diagnosis of IgG4-RD, a tissue biopsy may not be anatomically available depending on the affected organ(s) . The extent of fibrosis can predict poor responsiveness to immunosuppressive therapies –10. HoweThe enhanced liver fibrosis (ELF) score is both a clinically validated surrogate score of the severity of tissue fibrosis and a predictor of clinical outcomes in the setting of chronic fibrotic liver diseases . The ELFIn addition to CCL18 and the ELF score, growth differentiation factor 15 (GDF-15) and CCL2 are thought to be involved in the fibrotic manifestation of human diseases such as SSc . Serum lWe enrolled the 72 consecutive untreated patients who fulfilled the comprehensive diagnostic criteria for IgG4-RD  or organNo patients with hepatic involvement by IgG4-RD or chronic fibrotic liver disease were enrolled. Sixty patients underwent a biopsy, and the diagnoses of 51 patients were confirmed by histopathology . In 12 other patients, a tissue biopsy was not anatomically available. None of the patients was on GC treatment at baseline. Thirteen patients  were also evaluated after GC treatment.This study was a retrospective observational study using anonymized information. The patients gave their informed consent to be subjected to the protocol, which was approved by the institutional review board of each institution.Baseline serum samples were collected from untreated 72 patients at the time of diagnosis. Serum samples at sustained remission after induction of GC treatment were collected from 13 patients. Serum samples were aliquoted and stored at − 20 °C until analysis. The serum levels of GDF-15 , CCL2 (MCP-1) (R&D Systems), TIMP-1 (R&D Systems), and HA (R&D Systems) and PIIINP  were measured with specific enzyme-linked immunosorbent assay kits. The assays were calibrated using standards provided by the manufacturers. These biomarkers were also measured in 44 healthy controls . The ELF score provides a single value obtained with an algorithm combining the quantitative serum measurements of TIMP-1, PIIINP, and HA , 29. TheU test for comparisons between independent medians, and we used the chi-squared test for the evaluation of the associations between categorical variables. Correlations were assessed with Spearman’s correlation coefficient. The changes in the serum GDF-15 concentration from baseline were analyzed by using the Wilcoxon signed-rank test. We investigated the organ involvements associated with high GDF-15  by performing a multivariate logistic regression analysis. Variables with P values of less than 0.05 in a univariate model were used in the multivariate model. The overall significance level for statistical analysis was 5% (two-sided). P values of less than 0.05 were accepted as significant.Statistical analysis was performed by using JMP Pro statistical software, version 11.0 . Quantitative variables are presented as medians and interquartile ranges (IQRs). Categorical variables are presented as percentages. We used the Mann–Whitney n = 49 cases, 68.1%), lymph nodes , lacrimal glands , pancreas , retroperitoneal fibrosis , lungs , parotid glands , kidneys , orbits , prostate , bile duct , thyroid , aorta , and pituitary gland . A median of three (range of one to eight) organs per patient were involved. Fifty-two (72.2%) patients had at least three organs involved.The demographic and clinical characteristics of the 72 patients with IgG4-RD are summarized in Table The median (IQR) of serum IgG4 concentrations was 457 (260–705) mg/dL, and the concentration was at least 135 mg/dL in 68 patients (94.4%). Regarding serum complement levels, low C3 (<65 mg/dL), low C4 (<13 mg/dL), and low CH50 <30 U/mL) concentrations were seen in 15.2%, 24.6%, and 19.4% of the patients, respectively. Two or more serum complements were depressed in 13 patients (18.1%), and kidney was an involved organ in seven of these patients (53.8%). The serum concentration of soluble interleukin-2 receptor (sIL-2R)  was elevated in 34 of the 58 patients tested (58.6%), and the serum concentration of IgE  was elevated in 35 of the 60 patients tested (58.3%). The median (IQR) of IgG4-RD responder index (IgG4-RD RI) score at baseline, calculated according to Carruthers et al. [ U/mL conFifty-seven patients (79.2%) initially received treatment with GCs depending on the treatment guidelines for each organ involvement, and no patients were treated with any other therapies. The median initial dose of GC was 30 mg daily. Other patients were followed without any treatment.P <0.0001, Table The serum concentrations of GDF-15, CCL2, TIMP-1, HA, and PIIINP were significantly higher in the patients with IgG4-RD compared with the healthy controls  were positively correlated with each other Table . The ELFThe patients’ serum concentrations of GDF-15, CCL2, HA, TIMP-1, and PIIINP and the ELF score were not correlated with the IgG4-RD RI score or the number of organ involvements (Table P = 0.026) and the presence of parotid gland involvement  were independently associated with high GDF-15. Since GDF-15 was high in male patients compared with female patients (P = 0.0008) and the positive correlation of GDF-15 with age was present , a multivariate logistic regression analysis was adjusted by age and gender which still showed the tendency .We attempted to identify the organ involvements associated with a high GDF-15 level  by performing a multivariate logistic regression analysis . The serum concentrations of GDF-15, CCL2, and TIMP-1 were increased. The serum concentrations of HA and PIIINP as well as the ELF score did not change during GC treatment.The changes in the levels of serum biomarkers after induction of GC treatment in 13 patients are summarized in Additional file In regard to the serum cytokines and chemokines of patients with IgG4-RD, the associations of CCL18  and the As reported previously, we observed that the ELF score was increased in patients with IgG4-RD. We also observed that the serum concentrations of GDF-15 and CCL2 were significantly high in the patients compared with the healthy controls. The serum concentration of GDF-15 was the most useful for distinguishing patients with IgG4-RD from healthy controls. This is the first report of associations of GDF-15 and CCL2 with IgG4-RD. There is a close relation between GDF-15 and CCR2. GDF-15 upregulates CCL2 and its receptor CCR2 expression in macrophages and promotes macrophage chemotaxis , 19, sugA recent study demonstrated that the CCL2–CCR2 axis regulates the macrophage polarization of M2 dominant by influencing the expression of functionally relevant and polarization-associated genes and down-modulating pro-inflammatory cytokine production . FurukawOur present findings revealed the association of both GDF-15 and CCR2 with IgG4-RD, but there are remaining clinical outcomes that could not be explained by GDF-15 or CCR2. First, although the serum concentration of GDF-15 was correlated with some organ involvements of IgG4-RD, we observed that the conventional serum markers, including IgG4, complements, and sIL-2R, were correlated with the IgG4-RD RI score or the number of organ involvements better than GDF-15 or CCL2. Second, a clinical response to prednisolone was shown by our data, but the serum concentrations of GDF-15 and CCL2 were even elevated in the prednisolone-treated patients. It was reported that B-cell depletion therapy with rituximab reduced the IgG4-RD RI score, ELF score, and myofibroblast activation . The pre+ cytotoxic T cells [In addition to serum biomarkers, changes in the numbers of circulating plasmablasts , 34, act T cells  were obsWe observed increased serological surrogate outcome measures of fibrosis in patients with IgG4-RD. GDF-15 may precisely reflect the fibrotic degree in patients with IgG4-RD.Additional file 1:Table S1. Association between growth differentiation factor 15 (GDF-15) and organ involvements. Table S2. Changes of serum biomarkers and immunoglobulin G4-related disease responder index (IgG4-RD RI) score after glucocorticoid treatment in 13 patients. (DOCX 18 kb)


Intraoperative cultures are commonly sent in complicated appendicitis. Culture-guided antibiotics used to prevent postoperative infectious complications are debated. In this study, we describe the microbial overlap between intraoperative and abscess cultures, and antibiotic resistance patterns.A local register of a children’s hospital treating children 0–15 years old with appendicitis between 2006 and 2013 was used to find cases with intraoperative cultures, and cultures from drained or aspirated postoperative intraabdominal abscesses. Culture results, administered antibiotics, their nominal coverage of the identified microorganisms, and rationales given for changes in antibiotic regimens were collected from electronic medical records.In 25 of 35 patients who met inclusion criteria, there was no overlap between the intraoperative and abscess cultures. In 33 of 35 patients, all identified intraoperative organisms were covered with postoperative antibiotics. In 14 patients, organisms in the abscess culture were not covered by administered antibiotics. Enterococci not found in the intraoperative culture were found in 12 of 35 abscesses. We found no difference in the antibiotic coverage between rationales given for antibiotic changes.The overlap between intraoperative cultures and cultures from subsequent abscesses was small. Lack of antibiotic coverage of intraoperative cultures was not an important factor in abscess formation.The online version of this article (10.1007/s00383-018-04428-3) contains supplementary material, which is available to authorized users. In patients with perforated appendicitis, surgery and antibiotics are used to limit bacterial contamination. Commonly, broad-spectrum antibiotic regimens are used to cover the most common fecal bacterial pathogens that are released into the peritoneal cavity once the appendix perforates. Findings of the same bacteria in distant abscesses and periappendiceal abscesses were described in patients with appendicitis by Altemeier in the 1930s ), and two patients with B. fragilis or C. albicans were identified.One important group to consider is patients where IAA organisms were covered by antibiotics according to the resistance pattern, but appeared in the abscess anyway. Out of the nine such cases, four had C. albicans was found in IAA cultures in three patients , and Saccharomyces cerevisiae was found in one patient . Only in one patient with C. albicans an antifungal was used, reflecting that not all organisms were considered clinically relevant.Enterococci not found in the intraoperative culture were found in 12 of 35 IAA cultures and were, as a group, the most common organisms in IAAs not found in intraoperative cultures. S. anginosus was found in 18 (51%) and 6 (17%) of intraoperative and IAA cultures, respectively. Out of the six patients with S. anginosus in the IAA, five also had S. anginosus in the intraoperative culture. Cultures with ESBL E. coli were found intraoperatively in three patients, and in one of these  ESBL E. coli was also found in the abscess. The latter strain was susceptible in vitro to meropenem which was the antibiotic given before draining the IAA.S. anginosus strain found intraoperatively was sensitive to TMP/SMX, but resistant to TMP/SMX in the IAA. In the second patient , E. coli was found resistant to TMP/SMX, but a sensitive E. coli was found in the IAA. The third patient  had more intricate changes, with S. anginosus and B. fragilis found sensitive to clindamycin, but resistant in the IAA. The same patient had B. fragilis initially intermediately sensitive to piperacillin/tazobactam, but fully sensitive in the IAA. The fourth patient  had intraoperative B. fragilis sensitive to piperacillin/tazobactam, and intermediately sensitive B. fragilis in the IAA. Overall, these changes in susceptibility did not cause any difficulty in covering the respective organisms with the postoperative antibiotic regimens used, and in only one case possible resistance to a given antibiotic developed.Changes in antibiotic susceptibility between intraoperative and IAA cultures were only found in four patients. In one patient , the Enterococci, or by organisms in vitro sensitive to the antibiotics administered resulting in no growth in cultures from the IAA. Changing antibiotic susceptibility between perforation and abscess formation was not found to be an important factor for abscess formation.We found that the overlap between intraoperative cultures and IAA cultures was limited, with only 29% (10/35) of patients having some overlap in cultured organisms. Although 91% (32/35) of intraoperative cultures were covered by given antibiotics, only 40% (14/35) of patients had IAA organisms resistant to given antibiotics. The main mode of abscess development seems to be from resistant organisms not dominant in the intraoperative culture, typically The group without changes in the postoperative antibiotics probably had a clinically less severe postoperative course, as changes in the antibiotics were not attempted, and is corroborated by the shorter time in hospital in this group. It can also be speculated that empiric alterations in the antibiotic regimen were motivated by clinical deterioration and could constitute a sicker group. However, this is not supported by the data, where instead the patients with guided changes appeared to have stayed longer in the hospital, although the difference was not statistically significant.S. anginosus tended to be present also in intraoperative culture (5/6), whereas Enterococci were found de novo in the abscesses. When S. anginosus is present in the intraoperative swab, previous studies indicate that abscess formation is overrepresented [S. anginosus is overrepresented in complicated appendicitis [Enterococci in the IAA are worrying in that they are not necessarily covered by initial antibiotic regimens, a natural resistance to many common antibiotics, and because of an increase in vancomycin-resistant Enterococci (VRE). Our current antibiotic regimen in complicated appendicitis has now changed again to include piperacillin/tazobactam, with better coverage of, at least, E. faecalis.Two patterns that we noted regarding the most common organisms in IAAs were that resented , 12, 13,ndicitis . We beliThere is some data to suggest that not covering the organisms found at the time of surgery is associated with increase in superficial wound infections and a non-significant increase in abscess formation  and someAn additional obstacle in reviewing the effect of peritoneal swabs is that the pathogens causing postoperative IAAs are not characterized unless abscesses are found and drained or aspirated, and that intraoperative antibiotics are usually already administered reducing the sensitivity of the culture. There is also no clear consensus on where the most representative culture should be sampled in the contaminated abdomen to best predict how to prevent abscess formation.S. anginosus, could be reasonably settled with the clinically based assumptions described here. A further limitation is that only 77% of patients were cultured intraoperatively. In the subgroup at highest risk for IAA, i.e., those with perforated appendicitis, 94% were cultured. We find non-adherence to the local recommendation of culturing all appendicitis cases to be an unlikely source of serious bias given the dominance of perforated appendicitis cases in this IAA cohort. There was considerable variation in the antibiotics given over this 8-year period and this may make the interpretation of the results more difficult. However, local recommendations were followed in the postoperative antibiotic given in most cases, and the strategy with respect to sending intraoperative cultures and letting them guide antibiotic changes to be largely divorced from the specifics of the antibiotic type as long as appropriate initial broad-spectrum antibiotics are used.A bias in this retrospective review of patients who developed IAAs is that we cannot compare how many IAAs were prevented by changes in antibiotics due to information from the intraoperative culture. Looking instead at all patients who underwent treatment for perforated appendicitis, previous work found increased infectious complications, longer fever duration, and longer hospitalization with tailored antibiotic changes compared with empiric changes . Again, Because antibiotic choice changes over time as resistance patterns change, it is important to find practice guidelines that help not only with what antibiotic to use, but with the rationale for changing and stopping antibiotics. Clearly, as we show, intraoperative cultures are not easy to interpret as they poorly reflect the contents of IAAs, and to narrow treatment to intraoperative findings could be misleading given the frequent occurrence of other organisms in IAAs. Empiric broad-spectrum coverage of the common pathogens is reasonable going forward, and in the light of this study and previous work the value of intraoperative cultures is doubtful.Below is the link to the electronic supplementary material.Supplementary material 1 (DOCX 88 KB)Supplementary Fig. A1: Antibiograms with organisms and antibiotic susceptibilities for the 35 patients with intraoperative  cultures (EPS 1772 KB)


This paper presents the experimental results on the behavior of fly ash geopolymer concrete incorporating bamboo ash on the desired temperature (200 °C to 800 °C). Different amounts of bamboo ash were investigated and fly ash geopolymer concrete was considered as the control sample. The geopolymer was synthesized with sodium hydroxide and sodium silicate solutions. Ultrasonic pulse velocity, weight loss, and residual compressive strength were determined, and all samples were tested with two different cooling approaches i.e., an air-cooling (AC) and water-cooling (WC) regime. Results from these tests show that with the addition of 5% bamboo ash in fly ash, geopolymer exhibited a 5 MPa (53%) and 5.65 MPa (66%) improvement in residual strength, as well as 940 m/s (76%) and 727 m/s (53%) greater ultrasonic pulse velocity in AC and WC, respectively, at 800 °C when compared with control samples. Thus, bamboo ash can be one of the alternatives to geopolymer concrete when it faces exposure to high temperatures. For developing countries, bamboo is considered as an ideal crop for rural development. Bamboo production and utilization are considered relevant to many in the UN for sustainable development goals. Naturally, the bamboo is found in cylindrical pole or culm. Bamboo is also part of the grass family. There are over 1200 species of bamboo all over the world, with structural species varying by locations. The different species can be categorized into three types of root systems: sympodial (clumping), monopodial (running), and amphipodial (clumping and running). According to the Food and Agriculture Organization of the United Nations (UNFAO), a total of 72% of land area in Malaysia is filled with forests. Bamboo is an easy plant to grow. Tropical rainforest areas found in Malaysia provide ideal growing conditions for the bamboo plant. The production of bamboo charcoal has increased and its applications especially in healthcare, cooking, water purification, and gardening have grown significantly . ConsequThere is lack of research on the use of bamboo ash in the construction industry. Therefore, in present study, we have experimentally investigated the properties and performance of fly ash geopolymer concrete incorporating with bamboo ash under elevated temperature. In addition, there is no previous study that has used bamboo ash as a construction material. This investigation includes the effect of physical appearances, compressive strength, weight loss, and ultrasonic pulse velocity loss after the desired exposure.In this study, fly ash (FA) as low calcium fly ash (class F) and bamboo ash was used. The chemical composition and particle size analysis for both materials will be discussed in the results and discussion section.Fly ash was obtained from Tanjung Bin, Johor, Malaysia. The bamboo ash (BA) was obtained from Lanchang, Pahang, Malaysia. First, collected bamboo ash was dried in the oven for 24 hours at 110 °C (± 5 °C) to ensure that there was no moisture available. Then, the bamboo ash was ground in an abrasion test machine for 6 hours to improve the fineness of the ash. Then, the bamboo ash was sieved through a 45 μm sieve in order to remove bigger size of ash particle and impurities. Only the fine bamboo ash passing through the sieve were collected following the standard size of Portland cement used in the mixing. The specific gravity of FA and BA was 2.20 and 2.05, respectively.The standards used to determine the properties of aggregates are ASTM C127 , ASTM C13, respectively. River sand was used in fine aggregates and obtained from a local source in Johor, Malaysia. Specific gravity, water absorption, and bulk density of the fine aggregate was 2.6, 0.7%, and 1650 kg/m3, respectively. To ensure the aggregates did not absorb alkaline solution during the mixing process, saturated surface dry (SSD) conditions for both aggregates were conducted. For this purpose, both coarse and fine aggregates were soaked separately with clean tap water. Then, the aggregates were placed on a plastic sheet until the surface became dry.In this research, crushed granite with nominal size of 10 mm was used as a coarse aggregate. The specific gravity, water absorption, and bulk density of the coarse aggregate was 2.7, 0.5%, and 1551 kg/mTM, Auckland, New Zealand was used in this research. The alkaline solution was prepared by mixing 10 M sodium hydroxide (NaOH) with sodium silicate (Na2SiO3). The activator to binder ratio was different starting from 0.40, 0.45, and 0.5. Optimization of the activator to binder ratio was carried on the basis of workability results. Alkali Sodium based activator purchased from QReCIn this research, Master Glenium ACE 8589  was used as a superplasticizer (SP). SP is a chemical admixture that was added to the concrete during the mixing process. It is also known as a water reducer. SP provides exceptionally good early strength development and maintains flowability for a considerable period of time. Behzad and Jay have studied the effect of different SPs on the workability and strength of fly ash based geopolymer and they have found that SP is effective in improving the properties of concrete, which are directly dependant on the type of activator and the SP .The preliminary mix design was optimized based on several factors. In this study, the mass ratio of binder to aggregates and coarse to fine aggregates was set to be 1. The mass ratio for coarse to fine aggregates and sodium silicate to sodium hydroxide was set to be 1 and 2.5, respectively, along with 3% superplasticizer. Huseien et al. , have fo3 binder content in alkali activated mortar for durability properties [operties  and theyoperties . During The determination for the workability of the geopolymer mortar was conducted in accordance with ASTM C1437 [The compressive strength test measurement was carried out on a 100cm × 100cm × 100cm concrete sample. This test was conducted according to ASTM C109-16 standard .The chemical compositions of bamboo ash and fly ash were acquired by XRF test . 100 grams of both fly ash and bamboo ash was sealed safely in a plastic bag before being tested.X-ray Diffraction (XRD: Rigaku Smartlab Diffractometer) of fly ash, bamboo ash, 100% fly ash and 5% bamboo ash + 95% fly ash geopolymer concrete after 7 days of curing were carried out from 2θ = 5–85° using Cu radiation (λ = 1.54182 Å) at 25 kV.Particle size analysis (PSA) was conducted to investigate the size distribution of the binder. This analysis was performed by laser scattering technique (Mastersizer 3000). This test was carried out using the wetting method where the particles were dispersed using distilled water to avoid an agglomerated condition.The surface morphology of both materials (bamboo ash and fly ash) as well as 100% fly ash and 5% bamboo ash+95% fly ash geopolymer concrete after 7 days of curing were conducted by scanning electron microscopy  operated at 15 kV.The UPV value of concrete samples was determined according to BS EN 12504-4:2004  after exThe fire endurance test was carried out in accordance with ASTM E119-12a  using auThe chemical composition of bamboo ash and fly ash were determined using XRF. Based on the results shown in 2, JCPDS = 88-2487) and mullite  in the XRD pattern , rosenhahnite i.e., calcium hydroxide silicate (Ca3Si3O8(OH)2, JCPDS = 83-1242), and sulfur trioxide   b. Potass76-0760) . A sharp2/kg for bamboo ash and fly ash particles, respectively.The particle size distribution of fly ash and bamboo ash was characterized by their D50 values as shown in Once the activator to binder ratio was reached at 0.45, the slump value increases compared to 0.40 for 100% FA (Csample) and 5% BA+ 95% FA (Asample). Based on the observations, the mix was quite homogenous and the mixing process was easier compared to for the 0.40 ratio. The workable mix with the highest flow value was obtained when the activator to binder ratio was 0.50 for both samples. In conclusion, the small percentage i.e., 5% addition of bamboo ash at a 0.45 ratio, gave a much smaller slump loss in Asample compared to Csample. However, with 0.40 and 0.50 ratios, the slump loss is 10 mm for Asample. Thus, we decided to perform another experiment in a 0.45 activator to binder ratio. The standard deviation in the compressive strength for different mix proportions was found to be 0.11 to 1.31 MPa. A very stable proportion was needed in order to obtain a very good concrete mix for construction purposes. The addition of 5% bamboo ash was shown to be the most suitable mixture to get a better performance in regards to early compressive strength compared to other mixtures.2 and Al2O3 are not fully utilized for geopolymer formation. Bamboo ash and activator are the materials that are involved in the synthesis of fly ash based geopolymer concrete. The pure geopolymer network actually consists mainly of Si, Al, and O with alkali Na+ or K+. In this reaction, all minerals did not participate in the geopolymerization process. To understand the formation of phases after 7 days of curing, XRD of Csample (100% FA) and Asample (5% BA + 95% FA) was performed and results are shown in High temperature i.e., heat exposure is one of the most important parameters which affects the surface characteristics, surface outlook, shape, and color of concrete. Although it does not give significant information regarding the distortion suffered by the concrete, it will give an immediate impression of the failure tendency of the concrete. It can be observed from the entire cooling regime at 200 °C that the color of the sample was grey for both Csample and Asample with a smooth and sharp edges. These characteristics are maintained up to 400 °C. At 600 °C, the light grey color is observed for air cooling (AC) with rough surface while water cooling (WC) exhibits yellowish grey with cracks appeared in Csample whereas in Asample there is no cracks. At 800 °C, the Csample develops heavy cracks throughout the surface in AC and WC, whereas Asample shows lesser cracks compared to Csample in both AC and WC. The change in the matrix structure is a consequence of the dehydration in the phases of the binder during exposure to different temperatures. It is manifested as changes in porosity and color, as well as physical defects such as the presence of cracks. It is matched with the results presented in The residual compressive strength of Csample and Asample at different temperature and cooling regimes is shown in According to a previous researcher , geopolyIn this research, it was proved that fly ash geopolymer concrete (Asample and Csample) does not complete a geopolymerization process until 28 days have passed. Within 200 °C exposure, both type of samples gain an improvement in terms of strength and also the matrix structure. These are associated with the reported changes in the value of compressive strength after exposure. It is seen that the water molecule is expelled from the geopolymer concrete during the presence of heat, which improves the strength of the concrete and also leads to the discontinuous nano-pores of the matrix. It is possible that not all water molecules were expelled due to high temperature, especially in a larger sample. Higher surface tensions in larger samples will dissipate moisture at slower rates compared to in smaller samples. Starting from 400 °C, 600 °C and 800 °C levels of heat exposure, the the compressive strength decreased for both type of samples. But in the presence of bamboo ash, the residual compressive strength of Asample tends to show a positive result at 800 °C where the strength is higher compared to Csample. This clearly shows that at 800 °C, the presence of bamboo ash contains potassium oxide, thus producing a significant compressive strength compared to Csample. If alkali metal oxides content is above a certain limit, it exhibits a high coefficient of thermal expansion. With the presence of bamboo ash at high temperature, the tendency of the oxides lead to change its shape to have a higher area and volume. Thus, more solid and packed molecules are formed and provide better structural components for the geopolymer concrete. . At 400 °C, the trends changed, after which the Csample experienced UPV losses of 13.3% (AC) and 14.0% (WC) compared to Asample, which experienced 12.9% (AC) and 16.1% (WC) UPV losses. At 600 °C, the loss trends continued to increase and achieved a maximum loss at 800 °C for both types of samples. 67.8% (AC) and 64.7% (WC) losses was achieved by Csample, while 42.8% (WC) and 45.1% (AC) were achieved for Asample. The Asample exhibited a smaller loss of UPV compared to Csample. Generally, the reduction of the velocity in the concrete was due to the deformation of the microstructure in the geopolymer concrete. A rise in the temperature increases the amount of air voids in the concrete samples. Thus, the transmission speed of sound waves decreased with the increase in the traveling time of the ultrasonic pulse transmission.The change in UPV value due to exposure to high temperature is depicted in The quality of the concrete can be classified based on The effect of elevated temperature on the weight loss of geopolymer concrete for all curing regimes is depicted in In addition, an increase in temperature and weight reduction lead to deterioration of the structural integrity of geopolymer concrete. Asample yielded a higher weight loss compared to Csample. Furthermore, weight loss was observed to be lower in WC compared to AC regime. This may be due to water being absorbed during the application of water-cooling to make sure the temperature of the concrete was under control.2 = 0.8949  at 600 °C with a rough surface and cracks that appeared in Csample during water cooling (WC), whereas there were no cracks in Asample.Csample and Asample gained approximately 36% and 39% residual compressive strength at 200 °C of exposure, respectively, whereas losses of approximately 62% and 41% from 400 °C to 800 °C, respectively, were found after using the air cooling regime. Asample exhibited the highest residual compressive strength at 800 °C of exposure compared to Csample for the AC and WC regimes. UPV values for both types of samples increased tremendously at 200 °C of exposure, which was concluded to be good in terms of concrete quality. The UPV values tended to decrease from 400 °C to 800 °C of concrete exposure. A greater loss was found at 800 °C of exposure and it led to a change in the stiffness and mechanical properties of geopolymer concrete.Asample shows a high confidence level and the best fitted value compared to Csample in UPV vs. weight loss results, which reveals that the velocity of heat passing through the concrete is slower.Based on the experimental results and discussion, the following conclusions can be drawn:


The paper presents experimental investigations concerning the influence of the cement type (CEMI 42.5 R Portland cement and CEMIII/A 42.5 N slag cement—with 53% granulated blast furnace slag) on the mechanical and transport properties of heated concretes. The evolution of properties due to high temperature exposure occurring during a fire was investigated. High temperature exposure produces changes in the transport and mechanical properties of concrete, but the effect of cement type has not been widely studied in the literature. In this paper, concretes were made with two cement types: CEMI and CEMIII, using basalt (B) and riverbed aggregates (RB). The compressive and tensile strength, as well as the static modulus of elasticity and Cembureau permeability, were tested after high temperature exposure to 200, 400, 600, 800, and 1000 °C. The evaluation of damage to the concrete and crack development due to high temperature effects was performed on the basis of the change in the static modulus of elasticity. The test results clearly demonstrated that permeability increases with damage, and it follows an exponential type formula for both types of cement.  Cements with granulated blast furnace slag are widely employed due to their lower carbon footprint, as a strategy for sustainable development in the field of construction. The use of ground granulated blast furnace slag (GGBFS), which presents an amorphous structure and shows pozzolan characteristics, in concrete as an additive has a positive effect on the properties of fresh and hardening concrete ,2. The uWhen equal amounts of cement and water binder (w/b) are used, concretes with slag content have a lower compressive strength at early ages and higher compressive strength at late ages than Portland cement . FurtherEnvironmental conditions and the temperature exposure during curing has a strong effect on concrete mechanical properties ,5. When The existing knowledge regarding the behavior of high performance concrete in a fire was recently reviewed by the RILEM Technical Committee HPB-227 , however−6/°C and 17.2 × 10−6/°C, respectively, which is 99.2% and 75.5% of the value obtained for Portland cement paste [According to Shumuye et al. , the addnt paste . HoweverMoreover, it can be concluded that the addition of aluminosilicate minerals like fly ash, ground granulated blast furnace slag (GGBFS), and silica fume (SF) can affect concrete behavior at high temperatures in a way that may produce spalling of heated concrete in material that is denser, and thus less permeable ,19,20. LCases of fires that took place in engineering facilities  have caused numerous fatalities, but also significant financial losses. During these fires an important loss of concrete in tunnel linings was observed. The load-bearing capacity of the structural elements was reduced due to the explosive spalling. The spalling may take different forms, from small concrete pieces chipping, known as the popcorn effect, to explosive behavior when larger pieces of concrete are separated from the concrete element with great energy ,22,23,24During heating, the permeability usually progressively increases ,28,29, eHence, the results available do not reflect all the relevant aspects of this topic, and additional investigation is required. The literature results cannot be compared to each other due to the fact that the mixes differ. A research programme was therefore proposed which would allow for a clear comparison of the influence of cement type on the mechanical and physical properties of concrete at high temperatures. For this we performed various tests on identical concrete mixes, for which the only changing factor was the cement. Therefore, the main goal of this work is to present the comparison of the changes in mechanical and physical properties of concretes made with two different cement types; CEMI and CEMIII. For all four concretes, the composition of cement paste, as well as the volume of cement paste and mortar, remained the same. Thus, the study reflected solely the cement type effect of Portland cement versus slag cement on the mechanical performances and permeability of concretes made with two types of aggregates: crushed basalt (B) and riverbed gravel (RB). For all the concretes tested, the amount of all components (cement paste and mortar volume) and aggregate type and nature, as well as the particle size distribution, was identical, apart from the type of cement. cT, ftT, ET) were evaluated after heating to temperature T (°C), which corresponds to the post-fire performance of concrete in situations where the assessment of material properties is required. In this specific situation, the residual permeability of concrete is also an issue because it governs all aspects of durability, and there may be a need for assessment when a decision must be made on the further use of concrete elements after a fire.This research investigates the mechanical performances and permeability of concretes made with different cements, to compare their reference mass transport capacities, strength, and stiffness after high temperature exposure. The reference values of permeability enable one to assess their potential for spalling in fire conditions, as denser and less permeable materials are prone to this behavior. Furthermore, the evolution of permeability with heating temperature was investigated, as well as the compressive strength and splitting tensile strength. Moreover, the stress strain curves were determined, and the modulus of elasticity was determined. All residual mechanical performances basalt or (RB) riverbed gravel.Cements from Lafarge  were used for both the CEMI 42.5 R Portland cement and CEMIII/A 42.5 N slag cement. The chemical characteristics of these cements are provided in Two types of aggregates were used in this research programme: gravel from Dunajec River  and crushed basalt.3/m3 and the mortar volume was 550 dm3/m3. The concretes are denominated as B CEMI, B CEMIII, RB CEMI, and RB CEMIII. Plasticizer  and superplasticizer  were used and the water-cement ratio (w/c) of the concretes was equal to 0.3.In All concrete cubic and cylindrical specimens were cast in plastic molds and stored for 24 h. After preliminary 24 h curing, the molds were covered with plastic lids for 7 days to prevent water evaporation. Samples were stored in laboratory conditions at T = 20 ± 5 °C and relative humidity HR = 50% ± 5%. Cylindrical specimens dedicated to permeability measurements were cut into discs with a diameter of 150 mm and thickness of 50 mm at the age of 28 days. At 90 days, all specimens for mechanical performance testing and permeability were heated in an electric furnace to T = 200, 400, 600, 800, and 1000 °C. As recommended by RILEM , a heatiThe permeability test used nitrogen as a gas media and the Cembureau method was applied . The tes3/s);Q: the measured gas flow intensity Q = V/t (m3)V: gas volume (mt: time (s)a: atmospheric pressure (1 bar = 105 Pa);PP: absolute pressure (Pa);2);A: cross section area of the specimen ;L: thickness of the specimen (m).In Equation (1), the permeability (k) was determined:The initial reference permeability of the concrete in the samples at 90 days old was determined on the specimens that were not pre-dried, in order to represent the non-dried condition in the real structure. Subsequently, the samples were heated to a temperature ranging from 200 to 1000 °C, and after cooling the permeability was measured. Each measurement value represented the mean value from three samples.The cubic specimens with side a = 150 mm were used for compressive strength determination, with a diameter (d) of 100 mm and height (h) of 200 mm for the cylindrical samples for the splitting tensile strength tests. Three samples were used to test unheated concrete and two were used to test heated concrete. The modulus of elasticity was determined from the stress–strain (σ-ε) using one cylindrical sample . All E values were expressed in GPa and calculated from σ-ε curves as the stress to strain and strain ratio in the range of 10% to 40% of the ultimate stresses. For all properties six temperature levels were studied: T = 20, 200, 400, 600, 800, and 1000 °C. The compressive strength test procedures applied were presented in EN 12390-3 , and theo20°C and permeability k, and the mechanical properties of compressive strength fc20°C tensile strength ft20°C and modulus of elasticity E20°C were determined after 90 days. The initial measurements, obtained for non-heated concrete properties, are presented in For B CEMI, B CEMIII, RB CEMI, and RB CEMIII concretes, the initial physical properties of bulk density ρThe progressive increase of the temperature resulted in free water evaporation and progressive dehydration of the material. The C-S-H, as well as portlandite and calcium carbonate decomposition, were progressive in higher temperatures. As a result, weight loss was observed and the progressive density changes were recorded. The bulk density of B CEMI, B CEMIII, RB CEMI, and RB CEMIII concretes decreased as a function of the temperature. The mean values of bulk density are presented in 3 and the B CEMIII 2533.2 kg/m3. The RB CEMI and RB CEMIII concrete were 2300.7 and 2315.6 kg/m3, respectively. Apart from the initial values of density observed in the non-heated pristine concrete, the changes of the density with the temperature were quite similar for both cement types.In tT are presented. Heating resulted in a progressive reduction in strength, nevertheless, the differences between CEMI and CEMIII concretes over a whole range of temperatures may be considered insignificant, in the scope of measurement error, or the scatter of results for this mechanical property. In As has already been shown in previous research, an important aspect in the high temperature behavior of concrete is the thermal stability of aggregates at high temperatures. This can be evaluated by thermo-gravimetric and differential thermal analysis, which indicate the physical or chemical transformation of aggregates. As has already been reported , basalt The stress–strain relationships for the tested concretes are presented in T) of heated B CEMI and B CEMIII, as well as RB CEMI and RB CEMIII, are shown in 20°C) were 44.4 and 48.9 GPa, respectively, for B CEMI and B CEMIII. For riverbed aggregate RB CEMI and RB CEMIII they were 30.6 and 29.7 GPa. These results show clearly that for concretes with the same volume of cement paste, the modulus of elasticity is related to the nature of the aggregate and is strongly related to concrete density. Higher values of ET were observed for both CEMIII concretes with RB and B aggregates.The static modulus of elasticity values :d to the mechanical damage. The term (1 – d) corresponds to 0(T0) is the initial value of the static modulus of elasticity, E0(T) is the modulus for mechanically undamaged material expressed in a function of heating temperature, and E(T) represents the static modulus of elasticity of mechanically damaged heated concrete. Previous studies ,35 have 20°C is the static modulus of elasticity tested at 20 °C and ET is the value obtained for heated concrete.Following this approach, in The damage factor follows a comparable increasing change for all tested materials and almost reaches the value of 0.9, which means that 90% of the concrete has deteriorated. However, at 400 °C the damage value becomes much higher for the basalt aggregate concretes in comparison with the riverbed aggregate ones. Overall, the damage values for the CEMIII concretes appear to be slightly lower than for the CEMI concretes, especially for the basalt-based materials. These changes may be qualitatively compared to the change of total damage with temperature of a high performance concrete . HoweverIt has already been noted that the changes to the inner micro-structure and permeability of the concrete may be characterized using this mechanistic approach, using damage evaluation to describe the high temperature degradation and/or micro-cracking effects ,26,37,380 the initial reference permeability, DE the damage factor, and CDE is the material dependent parameter, here equal to 8, which confirms the value obtained for another high performance concreteat elevated temperatures, but based only on the CEMI cement [DE value being equal to 8was obtainedfrom the regression curve with the coefficient of determination R2 of 0.86 value. Therefore, the proposed regression curve is limited in the range of temperature from 20 to 600 °C.Three points that do not follow the trend in In Equation (4) k is the permeability of the heated material, kI cement . The CDEType of cement influences compressive strength and permeability of 90 day concrete. Concretes with CEMIII presented lower permeability and higher compressive strength for both basalt and riverbed aggregate concretes;High temperature exposure strongly influences the mechanical and physical properties of concretes, and the damage to concrete increases with exposure temperature. A temperature increase leads to the reduction of strength and modulus of elasticity. The splitting tensile strength decrease is more pronounced than the compressive strength evolution.cT, ftT and ET, were very close or the differences were within the range of measurement error or the scatter of results of the properties tested;Minor differences between the mechanical properties of heated CEMI and CEMIII concretes were observed. The bulk density values, as well as the mechanical properties fThe nature of the aggregate has a dominant influence on the material physical density and mechanical properties of the tested concretes. The compressive and tensile strengths depend on the aggregate nature for temperature up to 400 °C; above this temperature level, similar values of strength are observed;The decrease in the mechanical properties is the result of progressive cement paste damage due to dehydration and chemical changes in the cement paste. Moreover, crack development due to the thermal mishmash of aggregate and cement paste results in nonlinear behavior of heated concretes;The course of changes of the relative value in the elastic modulus for all the concretes investigated was very similar, except for the temperature of 400 °C. The riverbed aggregate concretes RB CEMI and RB CEMIII hada lower damage parameter than that observed for basalt aggregate concretes (B CEMI and B CEMIII) at this temperature. For 200, 600, 800, and 1000 °C, the damage levels were similar;−17 vs. basalt 0.7 × 10−17; concretes with CEMIII: riverbed 0.99 × 10−17 vs. basalt CEMIII 0.53 × 10−17. That difference was be explained by lower permeability of basalt aggregate itself. This relation was also observed for the temperatures of 200, 400, and 600 °C;Important changes of up to six orders of magnitude were observed in permeability values following heating. However, the differences between the concretes could not be considered as significant. Indeed, CEMIII concretes presented slightly lower values of permeability in comparison with the CEMI ones in whole range of temperatures. On the other hand, basalt aggregate-based concretes have slightly lower permeability than riverbed ones. Concretes with CEMI: riverbed 1.2 × 10Analysis of the results allowed the formulation of the constitutive exponential law, presenting the relationship between the permeability of concrete and damage, which is valid up to 600 °C.It can be considered that heating induces damage, which may be represented by changes in the initial modulus of elasticity, that depends to a small degree on the type of cement. In this range of damage, the effects of aggregate type are also non-significant.This paper intended to present the study of the influence of the cement types CEMI and slag cement CEMIII, in which the GGBFS amount reaches 54%, on the mechanical and physical performances of heated concretes with riverbed (RB) and basalt (B) aggregates. Four concretes with the same volume of cement paste and mortar were investigated. The only parameter differentiating RB and B concretes was the cement type. Analysis of the experimental data obtained concerned the mechanical tests, stiffness, and the permeability test results of the four concretes subjected to high temperature exposure (up to 1000 °C). The following main conclusions were drawn: