Anthropometric measurements of the human body are an important problem that affects many aspects of human life. However, anthropometric measurement often requires the application of an appropriate measurement procedure and the use of specialized, sometimes expensive measurement tools. Sometimes the measurement procedure is complicated, time-consuming, and requires properly trained personnel. This study aimed to develop a system for estimating human anthropometric parameters based on a three-dimensional scan of the complete body made with an inexpensive depth camera in the form of the Kinect v2 sensor. The research included 129 men aged 18 to 28. The developed system consists of a rotating platform, a depth sensor (Kinect v2), and a PC computer that was used to record 3D data, and to estimate individual anthropometric parameters. Experimental studies have shown that the precision of the proposed system for a significant part of the parameters is satisfactory. The largest error was found in the waist circumference parameter. The results obtained confirm that this method can be used in anthropometric measurements. Anthropometric measurements of the human body are applicable to many aspects of human life . AnthropIn view of the above, it became reasonable to look for other measurement methods that could support or partially replace the existing techniques or tools. An alternative to classical anthropometry became imaging methods commonly used in medicine  , as wellIn the literature, there can be found some studies in which images from digital cameras ,15,16, KIn the group of systems and tools used to estimate the anthropometric parameters of the human body, the Kinect sensor has found a wide application. The device, characterized by a low price, and equipped with an RGB camera, an infrared camera, and an infrared emitter, has become a significant element of scientific research worldwide ,24,25.Among the publications describing the varied application of the Kinect sensor, there are studies in which the authors characterized the sensor in terms of hardware , softwarIt was also used as a tool supporting the process of estimating the anthropometric parameters of the human body. Some studies concerning this issue described the results obtained in systems based on one ,20,40,41He et al.  developeTo develop a system for the estimation of human anthropometric parameters based on the data from a depth camera;To develop a method for estimating anthropometric parameters from 3D scans;Using and verifying the possibility to estimate anthropometric parameters by the Kinect v2 sensor.The review of the literature shows that the application of depth sensors as a tool supporting anthropometric measurements of the human body may be justified, but there is a lack of comprehensive solutions that enable the measurement of many anthropometric parameters and are thoroughly tested on many objects. This study aimed to develop a system for estimating human anthropometric parameters based on a three-dimensional scan of the complete body made with an inexpensive depth camera in the form of the Kinect v2 sensor. The developed system builds a 3D human model based on the data obtained from the depth sensor, then performs the segmentation of this model and estimates seven anthropometric parameters featuring the human build. It should be noted that the Kinect v2 Sensor is used only to acquire depth data and can be replaced with another sensor  that makes it possible to obtain this type of data. Summarizing, the main contributions of this paper can be stated as:The research included 129 men aged 18 to 28. The men featured a weight at a level of 79.4 ± 11.7 kg and a body height of 180.2 ± 6.5 cm. All participants of the research gave their written consent to the anthropometric examination and consent to perform the 3D body scan.Body height (BH)—the body height was measured with a stadiometer  with an accuracy of up to 1 mm.Arm span (AS)—the subject stood with his back to the wall so that his back, buttocks, and heels touched the wall. The subject then raised both hands horizontally and the fingers of both hands were straightened. Then the left hand with straight fingers touched the corner of the room. The arm span was measured with tape from the corner of the room to a mark on the wall that corresponded to the end of the right hand.Waist girth (WC)—measurements of the waist circumference were carried out with anthropometric tape, an approximate midpoint between the lower margin of the last palpable rib and the top of the iliac crest.Hip girth (HC)—the hip circumference measured around the widest portion of the buttocks.Arm girth (AC)—the subject was in a relaxed standing position with the arms hanging by the sides. The girth of the arm is measured by the anthropometric tape positioned perpendicular to the long axis of the arm at the level of the midpoint between the corner of the acromion and the proximal radial head. The tape should be positioned perpendicular to the long axis of the arm.Thigh girth (TC)—the subject stands with his legs slightly apart and his body weight evenly distributed on both feet. The measurement was carried out using anthropometric tape in mid-thigh in a perpendicular plane to the long axis of the thigh so that the flexible tape does not indent the skin excessively.Calf girth (CC)—the subject stood with feet slightly apart and body weight evenly distributed. Measurement was made in place of the maximum circumference of the calf in the plane perpendicular to the vertical axis of the leg. The measuring tape has been wrapped so that it does not indent the skin excessively.All anthropometric parameters were measured according to the International Standards for Anthropometric Assessment (ISAK) procedures  and inclThe system for estimating human somatic parameters  consistsibraries .z coordinate is searched. The y coordinate of the y-axis is in the range , it is necessary to perform segmentation in order to separate individual body segments. The 3D scan of the human figure is divided into 9 parts : head, ubody see . Then, ter torso . The caly axis, while the arm span (AS) is calculated on the basis of the coordinates of the points with the maximum and minimum value for the x axis. The procedure for determining the remaining somatic features from the 3D scan is as follows:In order to calculate given perimeters, fragments of point clouds are separated from individual segments. These points are determined as follows:(a)x is in the range Arm girth (AC)—the place (point (b)y is in the range Waist girth (WC)—the place (point (c)y coordinate corresponding to the location of the hip circumference measurement (y coordinate is in the range Hip girth (HC)—the value of the (d)Thigh girth (TC) is calculated for points located in the middle of the thigh segment. The (e)z coordinate is searched. The y coordinate of the y coordinate is in the range Calf girth (CC)—at the beginning, the approximate place of circumference measurement is determined, for this purpose, based on the measurements of the test group, during which the distances between the knee and the calf girth measurement place and the calf girth measurement place and the foot, the Using the Convex Hull method, an ordered list of points is determined from the points projected onto the plane;The perimeter is calculated from the equation:With a segmented 3D scan, somatic features can be determined. The height (H) of the human figure is calculated on the basis of the coordinates of the points with the maximum and minimum value for the d) and relative difference , defined as the two standard deviations of the differences of the paired parameters, and the coefficient of variance (CV), expressed in percentage as the quotient of the standard deviation of the differences of the individual paired parameters by the mean, were determined. Additionally, Pearson correlations between measured and estimated values were calculated. Statistical analysis was performed in the GNU R software [Statistical evaluation of the significance of the differences was performed using the U Mann–Whitney test, taking software .The experimental study consisted of verifying the accuracy of estimating selected anthropometric parameters calculated using the presented algorithm. The values of the obtained parameters were compared to direct measurement, which was considered as a gold standard (GS). The estimated results, along with the errors, are presented in The study shows that the most accurately estimated parameter was the body height parameter for which tionship . CorrelaAnalysis using the Bland–Altman method  showed tIn order to visualize in detail the differences between the calculated values from the 3D scan and the measured values (gold standard), analysis using Bland–Altman charts was used . The vasThe comparative analysis carried out showed that the estimation of selected anthropometric parameters using the depth sensor generates acceptable errors. The main quality criterion adopted in the presented solution was the comparison with direct measurements of anthropometric parameters. Comparison of the obtained errors with errors generated by models presented by other researchers is not obvious due to the use of different criteria for evaluating methods. An interesting study was presented by Kahelin et al. (2020) , where tAnother paper that allows a direct comparison with the proposed method is that of , which pA paper that also used a Kinect camera as a tool to determine a 3D scan of the human silhouette was by Tong et al. 2012 . Tong anThe Kinect sensor is very often used as a tool to scan either the entire figure or selected body segments ,40,41,43This paper presents and tests a method for measuring selected anthropometric parameters using an inexpensive depth camera in the form of the Kinect v2 sensor. In order to evaluate the method, a statistical analysis was carried out in the form of a U Mann–Whitney test and Bland–Altman charts. Experimental studies have shown that the accuracy of the proposed system for a significant part of the parameters is satisfactory (Limitations of the work are related to the validation of the method. The proposed method is tested only for selected anthropometric parameters. Another limitation is the research group. Parameter estimation was performed for the male gender. The method was not tested in a group of women. The research also did not take into account non-standard cases, such as body deformities, missing or shorter limbs, etc. In such cases, the system may give incorrect results.Future work will be related to the development of new functionalities of the algorithm and the use of machine learning methods to classify body composition components and somatotype components. It is also planned to test the proposed method with other depth sensors, e.g., Azure Kinect. In addition to using a more accurate sensor, we will also work on improving the accuracy of the method itself, for this purpose various filtration and smoothing algorithms will be tested. Another important element of future work will be the inclusion of the female gender in the study.T. Krzeszowski, K. Przednowek: “Method for estimating somatic features, somatic indicators, somatotype components, somatotype and body composition components with the use of depth sensor”, Polish patent publication PL240075B1, 2022.


Fiber reinforced polymer (FRP) strengthening in circular columns is known to be more effective than in rectangular and square columns because of the uniform distribution of confining pressure. This study explores the effectiveness of using carbon-FRP anchors to improve the confinement of square reinforced concrete (RC) columns strengthened with FRP. Sharp corners in non-circular columns cause stress concentration on the corners, reducing the effectiveness of strengthening. To address this, the study examines the impact of three different anchor configurations on two sizes of FRP-strengthened square columns. The results show that the proposed anchors distribute stresses to a greater extent, resulting in a more uniform distribution of stresses and better confinement. For the best performance, it is proposed that the anchor fans surround the corners of the cross section. Experimental findings and finite element analysis results using the Concrete Damage Plasticity model in the ABAQUS material library match. It is widely recognized that concrete buildings constructed in the past do not perform satisfactorily under extreme loading conditions . In addiConcrete column confinement with FRP sheets has provided a considerable increase in axial ,16,17,18Various parameters of FRP strengthening systems were tested in previous research, such as the amount and orientation of the composite fibers , the conTypically, circular sections result in confining forces or pressure producing a uniform stress distribution along the concrete section perimeter, while non-circular sections have different stress profiles due to the inability of the confining material to restrict displacement outside the central region of the concrete material. To reduce the problem of non-uniform stress distribution in non-circular columns, researchers have focused on modifying the concrete cross section. This can be achieved by chamfering the corners to a desired radius  or by adThough increasing the corner radius provides better FRP confinement, it is limited by the presence of existing reinforcement, as early code provisions call for smaller concrete covers. In addition, FRP usage is restricted for lower-grade concrete  according to design codes, respectively ,29. An aThis study proposes carbon fiber reinforced polymers (CFRP) anchors in low confinement areas to enhance the efficiency of FRP strengthening in non-circular columns. The system evenly distributes stresses across the column, improving confinement during concrete core dilation and increasing axial capacity. Bonding between anchors and concrete using epoxy offers significant tensile capacity and redistributes forces within the section. This study investigates the effects of three anchor configurations on FRP-strengthened square-section columns of varying sizes through experiments and finite element (FE) modeling with ABAQUS, version 2016.The experimental program aims to investigate the effects of varying parameters on the behavior of confined concrete columns. The parameters include the specimen size, anchor layout, and number of anchors . One CFRAn increase in the corner radius of the concrete specimens is known to enhance the efficiency of confinement. Literature data suggest that the minimum corner radius for full-scale concrete columns should be 13 mm . In thisThe anchors consist of a dowel, which transfers stress to the concrete core, and a fan that accumulates fiber stresses within the embedment depth and transfers them to the concrete core. The dowel part of the FRP anchor effectively simulates the confining behavior exhibited by steel hoops in reinforced concrete. Three types of FRP anchors were used in this study: simple anchorage (SA), corner anchorage (CA), and fan anchorage (FA) ,34. SA aTo prepare the anchors, FRP sheets of different lengths and 10 mm width were rolled, and the fibers were separated for better distribution along the surface. For dry anchor dowels, thin copper wires were wound around the fibers and secured with plastic clips to create a tube-like structure that can be inserted into the drilled holes .The general properties of the anchors are given in To minimize the scattering of test results due to the variation in concrete strength, a cubic compressive strength of 25 MPa was selected, and ready-made concrete was used to cast the specimens, all of which were made using the same batch and mix design .To achieve the desired rounded corners, modified plywood molds were used . Gypsum The adhesive used was a thixotropic, solvent-free, two-component impregnation resin based on epoxy with a tensile Young’s modulus of 4500 MPa (from the producer data sheet). Three concrete cubes (15 × 15 × 15 cm) and cylinders  were tested on the same day as the FRP-wrapped specimens to compare the unconfined and confined compressive strengths. After 28 days, the mean compressive strength of the confined concrete cubes was 29.1 MPa. Initially, the surface of concrete samples was treated with a concrete grinding machine to improve the adhesion of epoxy to the concrete. After grinding, the aggregate surface was visible, revealing the underlying voids, which were filled with epoxy. Holes were drilled on every specimen with a particular layout to place the anchors. The holes were chamfered with a needle scaler to avoid 90° bending of FRP anchors. The anchorages were placed in the center of the concrete column cross-section in order to evenly distribute the stresses from the FRP throughout the column. This helps to prevent the column from cracking or failing under load. The reinforcement procedure is shown in The shaded area shown in The FRP sheet was placed over epoxy using the hand layup method. The dowel was inserted through the FRP sheet and epoxy hole with minimal disruption to the fibers . The ancAll specimens were tested under uniaxial compression until failure . The loaThe tests showed that all three-anchor schemas increased the axial capacity and improved the ductility of the confined concrete column samples. The results followed the same path for both small- and large-size specimens. Results in terms of stress and strain values are summarized in The experimental results demonstrated that CFRP-confined columns exhibited superior capacity performance compared to unconfined ones. Specifically, the axial capacity increased by 23.15% and 14.29% for specimens A-1 and B-1, with corresponding axial strain increases of 2.7 and 2 times, respectively. Further enhancement of compressive capacity was achieved by using additional anchors with CFRP wrap, with improvements ranging from 2.94% to 17.5%. Among all the specimens tested, those where the anchorage extended beyond the corner displayed the highest contribution to compressive capacity. Notably, fan anchorages demonstrated better confinement for square columns and exhibited a larger axial load-carrying capacity than corner anchorages. For the B and A test series, the capacity of the fan-anchored confined column showed a 17.5% and a 16.17% increase, respectively, in comparison to their un-anchored counterparts. Detailed results are presented in The reduction in compression capacity in double-anchored specimens revealed that disproportionate disturbance resulted in lower levels of capacity improvement. On the other hand, A-F-2 and B-F-2 samples had eight anchor holes as well. However, the inclined orientation of the anchors did not disturb the core section of the specimen , resultiThe CFRP jacket fractured mostly near the corner due to stress concentration in un-anchored specimens (A-1/B-1). Confined specimens had CFRP tension failure at mid-height of corners , causingThe smaller samples (A) failed in the middle, above or below the anchor dowel. The larger samples (B) had different failure modes depending on the type of anchorage. Samples strengthened with single and double simple anchorage (B-S-1 and B-S-2) failed at the corners, while samples strengthened with corner  or fan (B-F) anchorage failed in the middle. The different failure modes were due to the different fan lengths of the two sets of specimens. The simple anchorage contributed more to the failure mode for the A series, as it acted partially as a corner anchorage for the smaller specimens. In summary, the uniformity of the confining pressure increased with the anchorages, and it was more significant for the specimens strengthened with the corner anchorage for both test series.cu/fcc > 90%). In contrast, un-anchored specimens showed considerably lower ultimate loads when compared to their peak strength . All anchored specimens in this experimental study exhibited little or no strain-softening behavior  15 × 15 × 30 cm column specimens. To reduce model size, only one-quarter of each column section was modeled along its longitudinal axis. Symmetrical boundary conditions were assigned, and axial loads were applied using a displacement control method. Experimental results in this study show that the anchors were firmly attached to the concrete core and did not separate from it under any failure behavior. Therefore, it was concluded that they were rigidly attached to the concrete core. Previous research has shown that actively confined and FRP-confined concrete have different plastic behaviors. FRP-confined non-circular sections have complex and non-uniform stress variations, but experimental and FE results agree on the general behavior and stress–strain curves of FRP-confined columns . This stThe Young’s modulus was calculated using the ACI formula ,28 as shtf of the concrete was taken as in Equation (2)The uniaxial tensile strength cσ and cε are the compressive stress and strain, respectively, and pσ and pε are the experimentally determined maximum stress and its corresponding strain, which are obtained from standard cylinder tests. The concrete under uniaxial compression is described by the stress–strain relationship proposed by Saenz  and giveτ) and effective separation (δ). Elasticity is defined by nominal strain and tractions, using an elastic constitutive matrix (Equation (5)) to show the behavior of each traction component.epoxyG is the shear modulus of adhesive in GPa and ctf is the tensile strength of concrete in MPa.cG is the shear modulus of concrete in MPa, epoxyG is the shear modulus of epoxy in MPa, epoxyt is the epoxy thickness (1 mm), ct is the concrete thickness (5 mm), cE is the Young’s modulus of concrete in MPa, and epoxyE is the Young’s modulus of the adhesive in MPa.The interface between concrete and CFRP was modeled using the cohesive zone model representation. A traction-separation model was used to represent the interface, assuming an initial linear elastic behavior with a stiffness K evolution of damage (Equation (4)). The model was interpreted in ABAQUS using a bilinear traction separation constitutive curve  in termsStress–strain curves obtained by FE analysis were plotted together with the experimental results in Overall, the FE analysis approach used in this study is in good agreement with the experimental results of anchored and un-anchored specimens. The damaged plasticity model shows better agreement with FRP-confined columns where anchorages were used.The influence of confining pressure and anchorage on axial stress capacity and stress distribution was investigated, with hydrostatic pressure playing a significant role. It was observed that failure occurred on the plane above the anchorage, which was analyzed subsequently. The sectional distribution of confining pressure in un-anchored and single-anchored specimens is shown in The S33 values in the legend box in This study aimed to reduce confining stress concentration in corners of FRP-confined prismatic column sections. To achieve this, FRP anchors were positioned on the faces with the least confinement and extended to the corners to distribute the stress more uniformly. This study evaluated three distinct anchorage layouts on two sizes of square column samples. The anchors improved the load-bearing capacity of the columns and prevented premature failures of the corners.Compared to columns without anchorage, the axial compression capacity of confined column specimens was increased by up to 17.5% when FRP anchorage was used. It is noteworthy that a significant increase in compressive strength was observed in the areas where the anchorages surrounded the corner, such as with the fan and corner anchorage. This figure clearly shows that simple FRP anchor application does not improve axial capacity, nor does confinement like steel crossties in reinforced concrete. Rather, FRP anchors should surround the corners of the member.Only a slight increase in axial strength was observed for double simple anchorage samples. This result may have been affected by the presence of eight anchor holes in a small cross-sectional area, which produced a weaker concrete core. To better understand the confining mechanism of multi-anchorages, a larger-scale specimen should be tested.The use of anchors resulted in a reduction in strain-softening behavior. The column specimens anchored with different anchorage layouts showed almost no strain-softening behavior. The confinement kept the concrete core intact beyond the peak load until rupture. The majority of the anchored FRP-confined specimens failed in the mid-face, indicating that the failure mode was directly influenced by the stress distribution mechanism of the anchor. This mechanism redistributed stress and provided a uniform confining pressure around the section.The finite element analysis approach used in this study is in good agreement with the experimental results of both anchored and un-anchored specimens. The analysis results indicate that the anchorage provided a higher axial stress capacity and a more uniform stress distribution over the section. These findings are in line with the experimental data. It can be highlighted that the results of this study have a number of implications for practice. First, this study shows that FRP anchors can be used to improve the confinement of FRP-strengthened square columns, where chamfering on corners is limited by rebar. Additionally, the required number of confining layers can also be reduced by introducing FRP anchors.This study explores the effectiveness of using CFRP anchors to improve the confinement of square RC columns strengthened with CFRP sheets by wrapping. Sharp corners in non-circular columns cause stress concentration on the corners, reducing the effectiveness of strengthening. To address this, the study examines the impact of three different anchor configurations on two sizes of FRP-strengthened square columns. Three types of CFRP anchors were used in this study: simple, corner, and fan anchorage. Simple and corner types aim at providing better confinement to the concrete core, whereas fans generate minimum disturbance to the column’s concrete core, which is the main part to carry the axial load. An increase in axial load capacity ranging between 14% and 20% was recorded for concrete samples reinforced without anchors (only CFRP wrapping). The application of a fan anchorage improved further the axial load capacity of the concrete samples, up to 39% compared to the control unreinforced specimens. Another interesting effect of the application of CFRP reinforcement is the improved post-elastic residual capacity after cracking. While the axial load capacity of unreinforced concrete specimens reduced dramatically after the peak load, reinforced ones were able to sustain a residual capacity up to 90% of the maximum compressive load. This was particularly noted for concrete specimens where anchors were in place. The results also show that the proposed anchors distribute stresses to a greater extent, resulting in a more uniform distribution of stresses and better confinement. For the best performance, it is proposed that the anchor fans surround the corners of the cross section. To achieve this, CFRP anchors were positioned on the faces with the least confinement and extended to the corners to distribute the stress more uniformly. On the other hand, it should be avoided to disturb core concrete. In order to achieve that, inclined drilling of anchor holes is a good practice. The authors suggest extending the study to rectangular columns, in which confinement by FRP wrapping is less effective with respect to square columns.On the other hand, this study has a few limitations. Only one or two concrete samples were tested for each reinforcement and a control. This makes conclusions difficult. Concrete is variable, and a difference in the control and treatment can be attributed to this variability and not necessarily to the reinforcement. However, several similar reinforcement layouts have been tested, for a total of 20 compression tests, and all of them produced an improvement in the axial load capacity of the concrete columns. This is a clear indication of the effectiveness of the reinforcement methods. More tests will be necessary to assess the effect of the anchors. The finite element analysis conducted in ABAQUS solidified our understanding of the anchors’ impact in FRP-confined square columns, aligning well with experimental findings. The results from the numerical simulations played a crucial role in validating the enhanced load-bearing capacity and more uniform stress distribution achieved through the use of anchors. This validation was paramount in illustrating the reduction in stress concentrations around corners, an inherent issue in non-circular columns. The integration of these findings into our conclusions strengthens the case for implementing anchors in practical applications, ensuring a robust and well-supported set of design recommendations.In addition, the investigation only involved FRP-strengthened square columns. It would be interesting to investigate the effectiveness of FRP anchors on rectangular columns. Second, this study only tested two sizes of columns as well. It would be interesting to consider the effect of FRP anchors on real-scale columns. Despite these limitations, this study provides valuable insights into the effectiveness of using FRP anchors to improve the confinement of FRP-strengthened square columns. The results of this study suggest that FRP anchors are promising for further improving the performance of FRP-strengthened columns. Future research should focus on investigating the effectiveness of FRP anchors on different shapes and sizes of columns, as well as the long-term performance of FRP anchors.


The antibiotic retention removal was higher than 80.0%. The antibiotic removal performance is greatly affected by the effect of pH, ionic strength, molar ratio, and/or OXT concentration, as these parameters directly affect the electrostatic interactions between the polymer and the antibiotics. The diafiltration technique was shown to be highly efficient for the removal of OXT, with maximum removal capacities of 1273, 966, and 778 mg OXT g−1 polycation. In conclusion, it can be said that coupling water-soluble polymers to the diafiltration technique is an excellent low-cost way to address the problem of antibiotics in aquatic systems.The presence of antibiotics in aquatic systems in recent years has become a global environmental and public health concern due to the appearance of strains resistant to these antibiotics. Oxytetracycline (OXT) is a high-impact antibiotic used for both human and veterinary consumption, and it is the second most used antibiotic in aquaculture in Chile. Based on the above, this problem is addressed using a linear polymer whose structure is composed of aromatic rings and quaternary ammonium groups, which will help enhance the removal capacity of this antibiotic. To obtain the polycation, a radical polymerization synthesis was carried out using (4-vinylbenzyl)-trimethylammonium chloride as the monomer. The polycation was characterized via Fourier Transform Infrared spectroscopy (FTIR) and Nuclear Magnetic Resonance (NMR). The removal studies were conducted under different experimental conditions such as pH levels , ionic strength (0.0–0.50 mg L Piscirickettsia salmonis. Research has shown that recent mutations, such as SLGO94 and SLGO95, of this pathogen present a higher level of resistance to these antibiotics , and ammonium persulfate (APS 98%) used as radical initiator. All the solutions used in this investigation were prepared with deionized water type I and all the reagents were purchased from Sigma Aldrich; Darmstadt, Germany.The reagents used for antibiotic removal were oxytetracycline hydrochloride  for weekly standard preparation. Hydrochloric acid standards 0.1 mol LThe polyelectrolyte homopolymer poly[4-vinylbenzyl) trimethylammonium chloride] was synthesized via free radical polymerization. A quantity of 0.2 mol of monomer was dissolved in 40 mL of deionized water. Subsequently, 1.0 mol % ammonium persulfate (APS 98%) as initiator was added to the amount of dissolved monomer and degassed for 10 min under nitrogen atmosphere. Polymerization was carried out for 24 h at 80 ± 1 °C under nitrogen atmosphere. For its purification, the obtained product was dissolved in 2000 mL of deionized water and ultrafiltered with regenerated cellulose membranes with exclusion weight of 100 kDa  and working pressure between 55.0 and 65.0 psi. It was then lyophilized and subsequently characterized .Fourier transform infrared spectroscopy (FTIR): the polyelectrolyte, antibiotic, and polymer-antibiotic were evaluated by FTIR spectroscopy. To determine the functional groups, a Nicolet spectrometer equipped with a DTGS-KBr detector , was used and analyzed in the range of 400–4000 cm−1.1H-NMR):Nuclear Magnetic Resonance  to dissolve 20 mg of each sample separately. The analyses were performed using a Bruker Advance 400 MHz instrument- .Scanning electron microscopy (SEM): it was used to study the surface characteristics of the homopolymer and polymer-antibiotic by scanning electron microscopy (SEM) using a JEOLSEM-PROBE CAMECA model SU-30, equipped with an EDS detector-Tokyo, Japan.To evaluate the removal of oxytetracycline using poly(4-vinylbenzyl) trimethylammonium chloride) and determine the ideal conditions to improve its performance, the diafiltration technique was used . The eff−1 antibiotic, solutions of 30 mg L−1 were prepared in deionized water with molar ratios of 1:20 of antibiotic and polymer. The pH was adjusted using 0.1 mol L−1 NaOH or HCl, and experiments were conducted separately at pH values of 3.0, 5.0, 7.0, 9.0, and 11.0. The effect of ionic strength was evaluated by maintaining an OXT concentration of 30 mg L−1, a molar ratio of 1:20 (OXT: polycation-HPE) at the pH value that showed the best performance. Separate experiments were performed by varying the NaCl concentration at 0.025, 0.050, 0.100, 0.25, and 0.500 mg L−1.The washing method was used to evaluate the best conditions for pH, ionic strength, molar ratio (Antibiotic: polycation-HPE), and antibiotic concentration. The same procedure was followed for each parameter evaluated, maintaining a constant cell volume of 20 mL and collecting fractions of the permeate with the same volume until completing 10 test tubes per analysis . From a −1. The results obtained were plotted as F (Vp/Vc) Vs Rpol (%), where F is given by the filtration factor which is the result between the permeate volume (Vp) and the cell volume (Vc). While, Rpol (%) is the retention of the polymer expressed in percentages.Then, we proceeded to study the increase in the molar ratio while maintaining the same concentration of the antibiotic with the optimal parameters of pH and ionic strength. In separate experiments, we evaluated molar ratios of 1:1, 1:5, 1:20, 1:40, and 1:100 (OXT: polycation-HPE). Once we determined the pH, ionic strength, and molar ratio parameters that yielded the best performance in OXT retention, we proceeded to evaluate the increase in antibiotic concentration. We evaluated concentrations of 1.0, 5.0, 30.0, 40.0, 80.0, and 100.0 mg LThe enrichment method was used to determine the maximum retention capacity (MRC) of the polymer for OXT . Using tp < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.The data are presented as the mean ± standard deviation. One analysis of variance (ANOVA) with Tukey’s test was used to analyze the significant differences between samples. For all tests, the level of significance was set to * −1 corresponding to C-H bond stretching, 2925 cm−1 C-C bond stretching, a strong absorption band at 1482 cm−1 characteristic of the quaternary amino group (-N+(CH3)3), and from 1623–1425 cm−1 attributed to the C=C bonds of the aromatic ring. Additionally, a strong signal at 3416 cm−1 is observed, which is characteristic of the OH groups and is attributed to the hydration of the polymer.In 1H-NMR spectra for both the homopolymer and its monomer. Characteristic signals between 2.5 and 3.0 ppm are attributed to the protons present in the (-N+(CH3)3) group  polyelectrolyte homopolymer was synthesized from [(4-vinylbenzyl) trimethyl ammonium chloride] to be coupled with ultrafiltration membranes and used for the removal of oxytetracycline in aqueous solution, which is a commonly used antibiotic in the salmon industry. The polycation was analyzed by FTIR and NMR, confirming the presence of functional groups and the absence of vinyl proton from the monomer. In pH studies, OXT removal rates of up to 80.0% were achieved at pH 9.0 with polymer doses of 25.5 mg. In ionic strength studies, a decrease in removal efficiency was observed as the salt concentration in the medium increased, indicating competition between chloride ions and the anionic OXT for the active sites of the polycation. It was also observed that increasing the polymer concentration resulted in an increase in the retention capacity in terms of mg L


Maximum specific heat, on the other hand, was reported for plain concrete (R)—1.678 ± 0.001 MJ/m3 K. The increasing amount of waste from the vulcanization industry has become a serious environmental challenge. Even the partial reuse of the steel contained in tires as dispersed reinforcement in the production of new building materials may contribute to reducing the environmental impact of this industry while supporting the principle of sustainable development. In this study, the concrete samples were made of Portland cement, tap water, lightweight perlite aggregates, and steel cord fibers. Two different addition of steel cord fibers  were used. The samples of lightweight concrete based on perlite aggregate with steel cord fiber addition showed a significant increase in compressive (18–48%), tensile (25–52%), and flexural strength (26–41%). Moreover, higher thermal conductivity and thermal diffusivity were reported after incorporating steel cord fibers into the concrete matrix; however, the specific heat values decreased after these modifications. The highest values of thermal conductivity and thermal diffusivity were obtained for samples modified with a 2.6% addition of steel cord fibers and were equal to 0.912 ± 0.002 W/mK and 0.562 ± 0.002 µm This state affects the search for alternative building materials and the improvement of those already in use ,2,3. In 3 . Lightwe3 ,6,7.However, the increased thermal insulation properties of lightweight concrete are influenced not only by the number of air voids in the final matrix but also by the amount of aggregate used ,9,10. ThPerlite aggregate is used in concretes and mortar mixes containing, in their composition, food waste ,14 or re3) in a 3D printed concrete matrix.Steel fibers are the leader in concrete reinforcement due to the high tensile strength of steel ,20,21. AHowever, it is not necessary to use new fibers produced in the steel mill. Alternative steel fibers that are a by-product of recycling can be used . Based oWith the acceleration of industrialization, the research on cement-based composite materials has diversified ,34. Ultr3, were used in this study, which corresponds to 1.3% and 2.6%wt. of concrete, respectively. The presented paper fills the gap and responds to the need of the construction sector regarding the use of recycled materials (steel cord from recycled old car tires) in concrete.In order to answer this demand, this study focuses on the possibility of using steel cord fibers from recycling old car tires to lightweight concretes based on perlite aggregate and checking how it affects the material and the mechanical and thermal properties of the hardened and fresh lightweight concrete mix. The addition of steel fibers in two amounts, 22 and 44 kg/mIn this study, three lightweight concrete mixtures were used: the first reference mixture (R) and two mixtures with steel cord addition mixtures (M1 and M2). All three tested materials consisted of CEM 42.5R Portland cement, expanded perlite aggregate and tap water. Moreover, two composites, marked as M1 and M2, had steel cord addition of 1.3% and 2.6% wt. of concrete, which corresponds to volume fractions equal to 1.8% and 3.1%, respectively. The doses of fiber were tested in the laboratory, and the 2.6% wt. of the concrete dose was the maximum one that allowed the proper mixing process of concrete components (without the flocculation process occurring in the mixer). The final composition of the tested samples is presented in CEM 42.5R Portland cement  was used as a binder to prepare all concrete samples. Specific physical parameters of cement used in this study are shown in Expanded perlite was added to the concrete mix. It was artificially received from perlite ore, which grounded to the proper fraction in the first phase. The final perlite aggregate was sieved into individual fractions, and in this study, two of them were used: fractions 0 ÷ 4 mm and 4 ÷ 8 mm in a ratio 1:1.5. Each lightweight concrete was designed using the volumetric method (method of successive approximations), supported by laboratory tests and experimental tests, which does not specify the requirement for the granulometric composition of individual lightweight aggregate fractions, i.e., the percentage content of individual fractions in the aggregate curve.In order to prepare all concrete samples, tap water was used. The water was obtained from the Military University of Technology connector. Before the test, it had not been purified, distilled, or otherwise treated in the laboratory, and its chloride content of it was 28 mg/L.A steel cord obtained in the old truck tires recycling process was used as fiber see . The tirTwenty-four hours before manufacturing the samples, the aggregate was put into the bathtubs filled with tap water a. This pAfter this stage, the water was added to the mixture in a thin stream, and the mixture was mixed for another 10 min. Then, in the case of the modified recipes, M1 and M2, the additive in the form of steel cord fibers was gradually dosed so that no local densities were made, and the additive was evenly distributed in the mixture. After adding the fibers, these mixes were mixed for another 10 min. The reference mix was also mixed for 10 extra minutes to standardize the mixing time of all recipes and to allow all mixes to air themselves to the same level. The total time of the mixing process was 40 min. The concrete mixtures were then poured into the molds, which were previously covered with a thin layer of anti-adhesive oil to help with the removal of hardened concrete. The fresh mix was placed in a mold in layers, and each layer was compacted with the use of the vibrating table for at least 1 min. In cubic molds (150 × 150 × 150 mm) and in the beams (150 × 150 × 600 mm), the mixture was poured in two layers, but in the cylinder molds (150 × 300 mm), it was poured in three layers. After vibrating, the upper surface of the samples was smoothed, and the extra material on top c was remIn order to analyze the structure of components of the lightweight concrete mix based on perlite aggregate, the scanning electron microscopy (SEM) technic was used. To analyze the structure of cement, a 1 g portion of cement CEM I 42.5R powder was collected and put under the JSM-6610 microscope  in a high vacuum. The area of the sample surface was subjected to scanning of a focused and concentrated electron beam of a high voltage of 12,000 V or 20,000 V, depending on the magnification needed. The primary electron beam penetrated the surface layer of the cement and induced a signal in it, which was treated as an analyzed secondary electron (SE) signal. With its help, it was possible to visualize the surface of the sample, in this case, the cement powder.The microscope was also used for field emission scanning electron microscope (FESEM) tests performed on the perlite aggregate. As this technic allows us to obtain the ultra-high resolution imaging of earth elements, it is the optimal technic to test perlite aggregate that is derived from ore. A representative sample of perlite aggregate was chosen randomly and put under the microscope. The procedure was similar to SEM, but a lower voltage was used to perform the FESEM tests (5000 V).For the analysis of the steel cord fibers, a representative sample in the form of ten random fibers was selected and then photographed using a confocal microscope LEXT OLS4100 . The confocal laser scanning microscopy (CLSM) technic allowed for obtaining microphotography of fibers with increased optical resolution and contrast by blocking defocused light. These sharp images also allow for the measurement of the total length and thickness of fibers (in five spots). Afterward, the thickness of each fiber was calculated as an average value.3 was taken from each type. Concrete was taken straight from the mixer and poured into the Abrams cone  in three layers; each layer was compacted. This process was performed with the use of 25 times hammering of the fresh concrete with a steel bar (diameter of 16 mm and length of 600 mm). Additionally, the fresh concrete mixture was tested for air content using the pressure method [After finishing the mixing of each lightweight concrete mix, the consistency was determined using the cone fall test according to EN 12350-2:2019-07 . Each mie method . For thiAfter the curing process of concrete, the samples were taken out of the water and initially dried. The density of the concrete sample was determined according to EN 12390-7:2019-08 . It was The compressive strength was determined by testing cubes with dimensions of 150 × 150 × 150 mm using a MEGA3–3000kN–100S testing machine . The machine was set to increase the force from 0.2 to 1.0 MPa/s, and the entire test was carried out at a constant speed without any dynamic disturbances . The speYoung’s modulus was determined using an odometer model V-E-400 . The test was performed by EN 12390-13:2014-02  and carrIn order to perform thermal properties tests, the analyzer ISOMET2114  was used. Its measurements were based on the obtained temperature responses to heat flow impulses through the tested lightweight concrete on perlite aggregate with or without steel fibers. The probe of the device was equipped with a resistor heater which was applied to the edge of the sample and generated a heat flux. The probe with a diameter of 60 mm was used due to the dimensions of the tested samples, i.e., 150 × 150 × 150 mm.FESEM images of expanded perlite particles  show thaThe steel cord fibers that were part of the concrete matrix had an average thickness in the range of 0.32 0.37 mm, as shown by the CLSM measures performed in points 2–6 see . MoreoveThe results of the cone fall tests are presented in 3 was qualified for the D1.6 lightweight concrete density class, but concrete modified with the steel cord fiber scored a lower class, i.e., D1.8 [3, respectively.The results of air contents of samples of lightweight concrete based on perlite aggregate are presented in e., D1.8 . The aveIn regular and heavy concretes, the air content is determined based on the consistency class and density; however, this trend does not apply to lightweight concrete due to the high porosity of the aggregate see . Air parThe results of mechanical strength tests are presented in The compressive strength of fiber-reinforced lightweight concrete is higher, by about 15% for M1 samples and by about 48% for M2 samples compared to the reference sample (21.93 ± 0.24 MPa). This phenomenon was also confirmed by Christidis et al. , who tesThe splitting tensile strength of the reference sample was equal to 1.30 ± 0.02 MPa. The splitting tensile strength of the M1 and M2 samples was higher by about 25% and 52%, respectively. Moreover, the Madandoust et al. study  reportedThe effect of steel corn fiber addition on flexural strength is inconsiderable. However, Christidis et al.  determin3 addition of steel fiber of length in the range 50–60 mm and aspect ratio 80 and 100, flexural strength was higher by 8–16%. This is about a 10% lower increase than the one for the M1 sample with 22 kg/m3 of fiber content. The values differ due to the different shapes of the fibers and the different compositions of concrete mixtures.The results obtained in this study are similar to the results of other scientists for traditional concrete. Splitting tensile strength obtained by Mensah et al.  for the 3). For the approximately same content of fiber (M1 sample with 22 kg/m3 of fiber content), the compressive strength was 5.47–9.97 MPa lower. Moreover, it can be explained by the different dimensions and shapes of steel fibers.In general, fibers do not affect compressive strength; however, tests may indicate an increase in compressive strength with the addition of fibers . Chen etThe way of destruction of the selected sample of fiber-reinforced concrete in the compressive strength test was presented in In the case of the bending strength test for the reference sample, the fracture occurred axially at the point where the force was applied a. The frThe modulus of elasticity was 11.45 ± 0.05 GPa for the reference sample and 12.75 ± 0.05 and 14.15 ± 0.05 GPa for the M1 and M2 samples, respectively. It can be noted that Young’s modulus increased with an increasing amount of steel cord addition. This is due to the characteristics of the longitudinal elasticity of steel, which is 20 times greater than that of concrete, so the increase is so large despite the small addition of dispersed reinforcement in the form of fibers. Additionally, an analogy to the compressive strength results can also be seen. Young’s modulus is closely related to this property of concrete, and this study confirms that this dependence also occurs when using lightweight aggregate. Furthermore, Zeybek et al.  reported3K and 0.055 MJ/m3K, respectively, for samples M1 and M2, compared to the reference sample (1.678 ± 0.001 MJ/m3K).As a result of the addition of the steel cord fibers, the thermal properties of the final composite are lower due to the higher thermal conductivity coefficient for steel than for concrete . The infThe rising trend of thermal parameters of the samples is similar to the rising trend of the density. This is because both of these values are closely related. For concrete with the addition of steel cord, higher thermal conductivity and thermal diffusivity, and lower specific heat were obtained than the reference sample of lightweight concrete. This is due to the use of additional material in the concrete matrix, which is a good heat conductor. As a result, this property deteriorated for the final construction product. Moreover, the thermal conductivity increase for lightweight concrete is in agreement with traditional concrete increase and disagreement when it comes to specific heat and thermal diffusivity, where opposite trends were reported . The inc3 of steel cord fiber were used, respectively. The cross-sections also show a significant number of pores, which are marked in blue (only the larger pores in the concrete that were visible to the naked eye). Additionally, the naturally occurring pores in the light aggregate cannot be seen in photos.In addition, these samples were examined under the microscope at a magnification of 15 times and 45 times. It can be observed that the steel cord fiber showed good pull-out strength, and the concrete cracked around its surface before the fiber was pulled out . FurtherThis observation was an analogy to the results of other scientists. Madandoust et al.  noted thWith the increase in the steel cord fiber up to 10% wt. of cement content, the higher values of the cone fall of the fresh, lightweight concrete mixture based on the perlite aggregate were determined;The steel cord fiber addition did not affect the air content in the concrete mix; however, it changed its density class from D1.6 to D1.8;With the 1.3% and 2.6% (wt. of concrete) addition of steel cord fibers, the mechanical properties of lightweight concrete based on perlite aggregate increased: the compressive strength by about 1.18 and 1.48 times, the splitting tensile strength by about 1.25 and 1.52 times, and the flexural strength by about 1.26 and 1.41 times, respectively;Steel cord fiber addition affects the modulus of elasticity as it was greater by about 11% for M1 and 24% for M2 samples compared to the reference sample;For steel cord fiber-reinforced lightweight concretes, higher thermal conductivity and thermal diffusivity and lower specific heat were obtained compared to the base sample.This study aimed to investigate the possibility of using recycled steel cord fibers in lightweight concrete based on perlite aggregate. The two different fiber content were used in this study (1.3% and 2.6% wt. of concrete). The main results of this study are as follows:The use of steel fibers in combination with lightweight aggregate in concrete will primarily have a positive impact on the road industry through the production of new elements such as paving stones, curbs, and road slabs. The elements will be lighter, which will shorten the implementation time and reduce the amount of work necessary to complete the project. For this purpose, the proposed recipes should be, however, tested in terms of abrasion resistance, slip resistance, and frost resistance, which is to be examined in future work on these recipes.


JBMR Plus published by Wiley Periodicals LLC. on behalf of American Society for Bone and Mineral Research.Stopping treatment for osteoporosis with denosumab (Dmab) leads to a major and rapid loss in bone mineral density (BMD) and a risk of vertebral fracture. Subsequent treatment with bisphosphonate (Bp) does not completely prevent this bone loss. We carried out a prospective pilot study to find out whether the gradual dose reduction with denosumab could prevent this bone loss. We proposed a therapeutic protocol consisting in reducing the doses of Dmab to women treated with Dmab for postmenopausal osteoporosis. Six months after the last dose of Dmab 60 mg, the subsequent injection was performed with a reduced dose of 30 mg, and the month‐12 injection was a 15‐mg injection. BMD and serum C‐terminal telopeptide of type I collagen (CTX) were measured at the start of treatment with Dmab (T0), at the last dose with 60 mg (T1), and at 6 months (T2) and 12 months (T3) after the last 15 mg Dmab injection. We included 13 patients aged 68.7 ± 3 years, and treated with Dmab for 45.2 ± 5 months. At the lumbar spine, 39% of the initial gain in BMD was preserved 1 year after the last dose (15 mg). Conversely, at the hip, the bone loss at the end of the treatment reduction protocol was equivalent to the initial gain. The mean CTX level was 166 ± 152 pg/mL 6 months after the last dose , and 549 ± 425 pg/mL 12 months after the last dose . One patient presented two vertebral fractures, 8 months after the last dose of Dmab (15 mg). Gradual dose reduction of denosumab (30 mg then 15 mg) does not prevent bone loss in the hip and partially maintains the initial gain at the spine. © 2023 The Authors.  Consequently, multiple vertebral fractures have been reported after Dmab discontinuation. Recently, Sølling and colleagues used zoledronate (ZOL) in 59 patients treated for 4.6 ± 1.6 years with denosumab to prevent this bone loss. The effect was incomplete: whatever the mode of administration of ZOL, ie, 6 or 9 months after stopping Dmab or when the markers of bone turnover increased, bone loss was, respectively, −4.8%, −4.1%, and −4.7% 12 months after stopping Dmab, at the spine. Other studies have confirmed the partial effect of ZOL or other bisphosphonates.The off‐treatment extension studies with denosumab (Dmab) have shown a rebound in bone turnover markers and a parallel loss in bone mineral density (BMD) following discontinuation of Dmab.1 ConsCould a gradual decrease in Dmab doses mitigate the rebound phenomenon of bone remodeling and attenuate bone loss? We proposed a Dmab therapeutic dose‐reduction protocol in 13 patients treated for postmenopausal osteoporosis.Our objective was to find out whether a gradual reduction in the dose of Dmab reduces the rebound phenomenon and bone loss caused by the discontinuation of this drug.T‐score >−2.5 standard deviation (SD) at the two sites  at inclusion, and without osteoporotic fracture over the last 2 years. Twenty patients met the inclusion criteria and 13 accepted our protocol.A total of 120 with postmenopausal osteoporosis have been treated with Dmab in our center since the product was marketed. We studied the files of these patients and reconvened in consultation those whose treatment by Dmab had been prescribed for at least 36 months. We performed a dual‐energy X‐ray absorptiometry (DXA) scan on these patients. We included women with postmenopausal osteoporosis, treated for at least 3 years of Dmab, without bisphosphonate treatment prior to Dmab, with a This pilot, monocentric, observational study was approved by the Toulouse II CCP. All patients signed informed consents.Six months after the last dose of Dmab 60 mg, they received an injection of 30 mg, followed 6 months later by an injection of 15 mg  and total hip sites using DXA  by a single experimenter, at the onset of treatment with Dmab (T0), upon discontinuation (T1), 6 months (T2) and 12 months (T3) after the 15 mg injection.BMD was measured at the spine  measurements were performed at T2 and T3 Fig. . .Data was expressed as mean ± SD. Variation of bone mineral was expressed as a percentage of the initial values. We tested whether values had a Gaussian distribution (Shapiro‐Wilk test). Multiple groups comparison was performed by a one‐way ANOVA and Tukey's multiple comparison test (Gaussian distribution). Correlation between bone loss and CTX variation was assessed by Pearson correlation test (Gaussian distribution). A T‐score was −2.3 ± 0.1 and the mean total hip T‐score was −2.2 ± 01. The mean duration of denosumab treatment was 45.2 ± 5 months.The mean age of our population was 68.8 ± 5 years. Regarding prevalent fractures: three patients had a history of vertebral fracture; two patients had wrist fracture. The mean lumbar The mean gain in BMD during treatment with full doses of Dmab was +7.8% ± 4.4% at the spine and +4.0% ± 3.5% at the hip after 45.2 ± 5 months of treatment  there was a bone loss of −5.4% ± 4.0% at the spine and −3.0% ± 2.0% at the hip  Table .One of the thirteen patients had two lumbar vertebral fractures due to a minor strain, 8 months after the last 15 mg dose of Dmab. This patient was 77 years old. For this patient, we reinitiated Dmab.T‐score >−2.5 SD and the absence of fracture during the 2 years preceding the study.We proposed our study to patients whose fracture risk seemed low, based on densitometric criteria Our study shows that the progressive decrease of Dmab helps preserve close to half of the BMD gain obtained with Dmab to the spine, whereas this strategy is ineffective for the hip. found in 11 patients that BMD gains obtained during treatment with romosozumab for 1 year and Dmab for 2 years were maintained in 73% (spine) to 87% (hip) of patients, 12 months after receiving an infusion of ZOL (post‐Dmab discontinuation). However, 24 months after ZOL, BMD decreased by −8% at the spine. An observational study including 120 postmenopausal women treated with Dmab for 2–3 years were given a single infusion of ZOL 5 mg, 6 months after the last Dmab injection. Six to Thirty‐six months after ZOL treatment, BMD had decreased significantly by −3.3%, −2.2%, and −1.5%, at the spine, total hip, and femoral neck, respectively. Anastasilakis and colleagues reported increased lumbar spine BMD 12 months after an infusion of ZOL (+1.7%), but decreasing BMD toward baseline values after 24 months (−0.1%) in the ZOL‐treated women. Recently, Sølling and colleagues reported a 2‐year randomized, open label trial, including 59 patients with osteopenia. They administrated ZOL 6 months , or 9 months  after the last denosumab injection or when bone turnover had increased . Their patients had taken Dmab for 4.6 ± 1.6 years, a length of time comparable to our study. The bone losses recorded in their patients are also similar to those of our patients: lumbar spine BMD had decreased significantly by −4.8% ± 0.7%, −4.1% ± 1.1%, and −4.7% ± 1.2% 12 months after ZOL in the 6 M, 9 M, and OBS groups. The progressive dose reduction of Dmab helps preserve close to half of the BMD gain obtained in the spine, but is not effective for the hip.Our findings of BMD decrease with Dmab doses reduction can be compared to several works studying the efficacy of ZOL post‐Dmab, the results of which are fairly heterogeneous, probably depending on the duration of treatment with Dmab and whether or not the patients had taken bisphosphonates before Dmab. Horne and colleagues8 foun This dose increased lumbar spine and hip BMD, when compared to placebo, but to a lesser extent than higher doses. However, the 14‐mg dose was not able to induce a sustained reduction of CTX levels over the 6 months of the inter‐dose period.It should be noted, in our study, that an injection of 15 mg of Dmab is sufficient to prevent bone loss and maintains CTX at a low level in most patients. These findings are in line with a previous report assessing various Dmab regimen including 14 mg of Dmab every 6 months in postmenopausal osteoporosis.10 ThiThe limits of our pilot study are linked to the small number of patients and absence of a control group. The main input for the scope is the fact that it is the only prospective study on the effect of the progressive decrease of Dmab. Moreover, we selected patients with high expected risk of bone loss on discontinuation of denosumab because they had no treatment with bisphosphonates before Dmab, and Dmab duration was longer than 36 months.To date, no therapeutic protocol using bisphosphonates fully preserves bone gain after Dmab withdrawal. A progressive decrease in treatment preserves ~40% of the gain at the spine. More studies about Dmab tapering are needed and potential usefulness of combining bisphosphonates with low‐dose Dmab remains an unexplored area.Administration of 15 mg of Dmab is sufficient to prevent complete BMD loss and to keep CTX levels low. This approach could be an alternative when bisphosphonates are contraindicated following Dmab.Michel Laroche: Conceptualization; data curation; formal analysis; investigation; methodology; supervision; validation; writing – original draft; writing – review and editing. Guillaume Couture: Writing – review and editing. Yannick Degboé: Formal analysis; writing – review and editing.The authors declare no conflicts of interest.https://www.webofscience.com/api/gateway/wos/peer-review/10.1002/jbm4.10731.The peer review history for this article is available at Fig. S1. Correlation between crosslaps and bone mineral density variations from T2 to T3.Click here for additional data file.


Vasculitis does not usually induce glandular tissue lesions. Herein, we present a case of vasculitis with a distribution similar to that of immunoglobulin-G4-related disease (IgG4-RD), including that of the submandibular and lacrimal glands, and periaortitis.A 70-year-old man presented with shortness of breath. Laboratory findings revealed an IgG4 level of 191 mg/dL, negative antineutrophil cytoplasmic antibody test, and C-reactive protein level of 8.33 mg/dL. Magnetic resonance imaging of the head and computed tomography of the neck revealed bilaterally enlarged submandibular and lacrimal glands. Neck-to-pelvis computed tomography revealed bilateral infiltrative shadows in the lower lobes of both lungs, mass shadows in both lungs, and periaortitis of the abdominal aorta extending to the common iliac artery. Thus, the patient was diagnosed with IgG4-related respiratory disease and periaortitis/periarteritis. Prednisolone was administered at a dose of 35 mg (0.6 mg/kg daily). The dose was gradually tapered while observing the effects of the treatment. Imaging findings indicated an improvement and the C-reactive protein and IgG4 levels decreased, indicating a successful treatment course. However, after reexamination of the pathological findings, the diagnosis changed from IgG4-RD to vasculitis. One year after treatment initiation, the patient symptoms have stabilized.Vasculitis can present with lesions and pathological findings similar to those of IgG4-RD. The diagnosis of immunoglobulin-G4-related disease (IgG4-RD) and vasculitis has been increasing since the novel concept of IgG4-RD was established in Japan in 2011.; however, glandular tissue lesions are not considered a typical presentation of vasculitis. Herein, we report a case of vasculitis in the submandibular and lacrimal glands with periaortitis, which presented with a distribution similar to that of IgG4-RD.IgG4-RD frequently affects the pancreas and lacrimal and salivary glands. Vasculitis commonly affects the upper respiratory tract, lungs, kidneys, eyes, and peripheral nervesEthics approval and consent for this case report were waived. We have obtained written consent from the patient to present the case.A 70-year-old man developed a sore throat and cough while working. He also experienced shortness of breath. He visited a physician 14 days later. Pneumonia was suspected based on his chest radiograph findings; therefore, the patient was referred to another hospital for treatment. Upon admission, a chest computed tomography (CT) was performed which revealed masses in the lower lobes of both lungs. However, no improvement was observed after 1-week of treatment with tazobactam/piperacillin (4.5 g × thrice/day). The patient was referred to our department for further management. He had a history of smoking 20 cigarettes per day for 47 years (20–67 years of age).2 saturation, 96% on room air; and temperature, 36.7°C. The submandibular glands were palpable and approximately 2 cm in size. We did not identify any pulmonary vessel murmurs, abdominal vascular murmurs, or any other remarkable physical findings.On admission, his vital signs were as follows: blood pressure, 117/67 mm Hg; pulse rate, 80 bpm; respiratory rate, 18/min; OThe abnormal laboratory findings upon admission were as follows: The IgG4, IgE, rheumatoid factor, antinuclear antibody, and C-reactive protein (CRP) levels and erythrocyte sedimentation rate were elevated. All other blood investigation results, including the myeloperoxidase-antineutrophil cytoplasmic antibody (ANCA) and proteinase-3-ANCA levels, were within the normal reference range Table .Head magnetic resonance imaging and neck CT revealed bilaterally enlarged submandibular and lacrimal glands Fig. A and B. Ultrasonography of the submandibular gland revealed coarse hypoechoic areas Fig. .Nasopharyngolaryngoscopy revealed no abnormal findings in the paranasal cavity; thus, no biopsy was performed. the patient was diagnosed with IgG4-related respiratory disease and periaortitis/periarteritis.We performed a CT-guided biopsy of the mass in the left lung; histological analysis revealed dense lymphocytic and plasmacytic infiltration with fibrosis. Immunohistochemical analysis revealed numerous IgG4 + cells in the mass (≥50 cells/high-power field); in some areas, the IgG4+/IgG + cell ratio exceeded 40. Storiform fibrosis and obliterative phlebitis were also observed Fig. A–C. BasePrednisolone was administered at a dose of 35 mg (0.6 mg/kg daily). The dose was gradually tapered while observing the effects of the treatment. The subsequent improvement in imaging findings and decrease in CRP and IgG4 levels, indicated a successful treatment course. There was no specialist in IgG4-RD in our hospital. Thus, we reviewed the samples collected before diagnosis; we did not repeat the biopsy. The diagnosis was changed from IgG4-RD to vasculitis.The lung tissue revealed damaged vascular endothelium, some nuclei debris, and neutrophilic infiltration, suggesting vasculitis Fig. A. The alOne and a half years after treatment initiation, the symptoms have stabilized; therefore, the remission is being maintained with a small dose of prednisolone. If symptoms worsen in the future, we will consider the use of immunosuppressive drugs in combination with prednisolone.We identified 2 important clinical issues in this case. First, vasculitis can present with a distribution similar to that of IgG4-RD. Second, submandibular gland ultrasonography may be useful for distinguishing between IgG4-RD and vasculitis.; it discussed that ANCA-negative AAV is analogous to seronegative lupus or rheumatoid arthritis. It is diagnosed if the patient meets the criteria for AAV, but the serological tests for ANCA are negative. In Japan, GPA is diagnosed based on the diagnostic criteria proposed by the Japanese Ministry of Health, Labor and Welfare. Although this case was not definitively GPA, it could probably have been GPA based on these criteria. Additionally, it could not be classified as GPA according to the American College of Rheumatology classification criteria of 1990. However, owing to the histological compatibility with the CHCC criteria of microscopic polyangiitis and radiographic evidence of fixed pulmonary infiltrates present for >1 month, this case was classified as GPA based on the European Medicines Evaluation Agency vasculitis classification algorithm. The 2012 CHCC also states that when patients with GPA exhibit clinical and pathologic changes identical to those observed in respiratory diseases they should remain in the GPA category.The vasculitis in this case may have been ANCA-negative granulomatosis with polyangiitis (GPA). The concept of ANCA-negative ANCA-associated vasculitis (AAV) was advocated in the 2012 Chapel Hill Consensus Conference (CHCC)–11 The submandibular gland lesion in this case was pathologically similar to that of IgG4-RD; however, cell debris is not typically observed in IgG4-RD, and the IgG4/IgG cell ratio was low. Moreover, parotid involvement in vasculitis is rare.–21 Thus, the relationship between salivary gland involvement and vasculitis remains unclear. Additionally, the patient had a lacrimal gland lesion, which has been reported in patients with vasculitis.–24The patient had a lesion in the submandibular gland. Involvement of the submandibular gland in vasculitis is rare.–35 The mechanism of large-vessel involvement in GPA is yet to be elucidated. Histological and clinical observations in previous studies have suggested that large-vessel involvement belongs to the AAV spectrum rather than overlapping with other large-vessel vasculitis.,36A periaortic lesion was observed in this patient. The CHCC 2012 criteria classifies GPA as a small-vessel vasculitis. Aortic involvement in GPA is rare, and only a few cases have been reported.; however, high CRP levels can be a hallmark of IgG4-related periaortitis/periarteritis. The median CRP level of IgG4-related periaortitis/periarteritis is 6.72 (2.14–24.65) mg/dL. In this case, the CRP levels was 8.33 mg/dL; therefore, we misattributed this elevation to IgG4-related periaortitis. This is a diagnostic limitation of the present study.The CRP levels in patients with IgG4-RD do not increase significantly–21 GPA with periaortitis was not reported in any of these case reports. Table –35 Among these, only 1 case of GPA reported a glandular tissue lesion; proteinase-3-ANCA was positive in this case.Table To the best of our knowledge, this is the first report of vasculitis with a distribution of lesions similar to that of IgG4-RD, including the submandibular and lacrimal glands, and association with periaortitis. In this case, coarse hypoechoic findings were observed, which differ from the ultrasonographic findings in IgG4-RD; this was useful for the diagnosis. Currently, IgG4-RD cannot be completely ruled out based on ultrasonographic findings alone; however, diagnosis can be made in combination with serological findings. Our study findings need to be externally validated at multiple facilities.Submandibular gland ultrasonography may be useful in distinguishing IgG4-RD from vasculitis. Characteristic ultrasonographic changes in the submandibular gland of patients with IgG4-RD are as follows: hypoechoic areas in a nodal pattern with high vascularity and/or superficial hypoechoic areas in a reticular pattern. The sensitivity, specificity, and accuracy of ultrasonographic diagnosis of IgG4-RD are 100%, 83.8%, and 91.2%, respectively.Vasculitis can present with a distribution similar to that of IgG4-RD, and ultrasonography of the submandibular gland may be useful in distinguishing between IgG4-RD and vasculitis. Unawareness of this, could lead to misdiagnoses. Therefore, ultrasonography of the submandibular gland should be performed frequently. Further studies should be conducted to determine the occurrence of conditions mimicking IgG4-RD and whether routine submandibular gland ultrasonography may contribute to the diagnosis.We would like to thank the Japanese Association for IgG4-related Diseases for reexamining the pathological findings.Writing – original draft: Mamika Kuribayashi, Hidesato Odaka.Writing – review & editing: Susumu Takahashi, Takuo Tokairin, Hirokazu Kurokawa.


TP53 mutation and c-MET activation are frequent events in human HCCs. Here, we discovered that the simultaneous mutations in TP53 and activation of c-MET occur in ~20% of human HCCs, and these patients show a poor prognosis. Importantly, we found that concomitant deletion of Trp53 and overexpression of c-MET (c-MET/sgp53) in the mouse liver led to HCC formation in vivo. Consistent with human HCCs, RNAseq showed that c-MET/sgp53 mouse HCCs were characterized by activated c-MET and Ras/MAPK cascades and increased tumor cell proliferation. Subsequently, a stably passaged cell line derived from a c-MET/sgp53 HCC and corresponding subcutaneous xenografts were generated. Also, in silico analysis suggested that the MEK inhibitor trametinib has a higher inhibition score in TP53 null human HCC cell lines, which was validated experimentally. We consistently found that trametinib effectively inhibited the growth of c-MET/sgp53 HCC cells and xenografts, supporting the possible usefulness of this drug for treating human HCCs with TP53-null mutations. Altogether, our study demonstrates that loss of TP53 cooperates with c-MET to drive hepatocarcinogenesis in vivo. The c-MET/sgp53 mouse model and derived HCC cell lines represent novel and useful preclinical tools to study hepatocarcinogenesis in the TP53 null background.Hepatocellular carcinoma (HCC) is a deadly malignancy with high genetic heterogeneity.  Primary liver cancer is one of the most common malignant tumors worldwide. Hepatocellular carcinoma (HCC) is the predominant tumor entity, accounting for 75–85% of the cases . HCC is TP53 (p53) gene is one of the most critical tumor suppressors. The p53 protein encoded by TP53 is involved in various pathways to regulate multiple processes, such as metabolism, DNA damage repair, cell cycle arrest, and apoptosis. As in other cancer types, TP53 mutation is also one of the main genetic variations in HCC. Accounting for about 30% of HCC cases, TP53 inactivating mutations contribute to HCC initiation and progression [The gression –6. In adgression , 8. TherMET (c-MET) is a protooncogene that encodes the hepatocyte growth factor (HGF) receptor. Binding to HGF, c-MET activates multiple downstream targets, such as the RAS/MAPK and phosphoinositide 3-kinase (PI3K)/AKT pathways, to drive tumor invasion and metastasis [tastasis . Previoutastasis , 11. Howtastasis –14.TP53 mutations are more likely to show upregulation of the c-MET signature. Specifically, about 22% of the HCC patients have both c-MET activation features and TP53 mutations. Clinically, these HCCs are associated with poor patients’ prognoses. Therefore, we speculate that TP53 mutations can synergize with c-MET activation to induce hepatocyte malignant transformation. In this study, we established and characterized a novel murine HCC model induced by overexpression of c-MET and CRISPR-Cas9 mediated knockout of Trp53 (encoding mouse p53). In addition, we applied this model for therapeutic testing.Gene expression analysis of HCC specimens from The Cancer Genome Atlas-Liver Hepatocellular Carcinoma (TCGA) database suggests that samples with TP53 mutation are two frequent alterations reported in human HCC. To investigate the profile of these genetic aberrations, we analyzed the expression level of c-MET and TP53 mutation status based on The Cancer Genome Atlas-Liver Hepatocellular Carcinoma (TCGA LIHC) human HCC dataset [TP53 mutation include truncating mutations, splice mutations, inframe mutations, and missense mutations. The details about TP53 mutations are shown in Fig. STP53 mutant and TP53 wild-type samples. The results showed that the c-MET signature was enriched in TP53 mutant human HCC samples  in the TCGA dataset displayed the activated c-MET signature. In comparison, only 55% of TP53 wild-type HCCs (136/249) exhibited high c-MET expression  and c-MET/sgp53 liver tumors (n = 3). The heatmap of the gene list showed genetic dissimilarities between the two groups   Fig. S. The Kyo5) Fig. S. Interes5) Fig. S. The corTP53 deletion using the TCGA-LIHC dataset, which occurred in 23/360 HCC specimens  in c-MET-high/TP53-null HCC samples, among which 521 genes were also upregulated in c-MET/sgp53 mouse HCCs  derived from a c-MET/sgp53 HCC by serial passaging of tumor cells from mouse to mouse Figs. S and S10.P adj < 0.05) in MP cells (1578 genes overlapped with the MP tumors) , we also performed RNA-Seq experiments on MP cells. Normal liver tissues from FVB/N mice were used as the control. The DEGs analysis identified 4697 significantly upregulated genes  Fig. S. KEGG ans) Fig. S. Notablys) Fig. S.Cabozantinib is an FDA-approved drug for HCC treatment, and it inhibits HCC growth by targeting the activated c-MET pathway , 21. TheTP53 is one of the most frequent genetic events in human HCCs, we applied this unique murine HCC cell line to study drugs that have shown effectiveness against p53-defective tumors. Niclosamide and metformin have been reported to inhibit the growth of xenografts from p53-defective human cancer cells [TP53 null human cancers is lacking. We found that both drugs could inhibit MP cell growth in vitro . The results suggested that the TP53-null HCC cell line (Hep3B) has higher sensitivity to multiple MEK inhibitors treatment than p53-mutant and p53-wild-type human HCC cell lines  mutant metastatic melanoma [Based on the in vitro studies, we investigated whether the MEK inhibitor effectively inhibits melanoma . First, Altogether, these data underline the therapeutic potential of MEK/ERK inhibitors for treating p53-deficient HCC.TP53 mutations abolish the functions of the p53 protein, such as gene transcription, DNA synthesis and repair, cell cycle arrest, senescence, and apoptosis [TP53 database (https://tp53.isb-cgc.org/), only a few murine liver cancer models with engineered p53 are reported in the scientific literature   Fig. S. SignifiC per se , making p53 HCCs . These aFor p53 mutated HCCs, the current drug development strategies include restoring wild-type p53 conformation and transcriptional activity, inducing the degradation of mutated p53, and inhibiting the interaction of p53 and its negative regulatory factor MDM2. However, the heterogeneity of p53 mutations in tumors limits the use of these drugs. Thus, synthetic lethal effects can be a promising therapy for a wide range of functional p53 mutations in HCC. Here, our murine HCC model and the related HCC cell line with deletion of p53 could be a valuable preclinical model in exploring targeted treatments for p53-loss-of-function HCCs in vitro and in vivo.TP53 null mutations. Therefore, it is critical to identify a potential biomarker(s) of response to the effective therapy for HCCs. Thus, preclinical and clinical studies are required to examine the therapeutic efficacy of trametinib against TP53 null human HCCs.The current study reveals that cabozantinib and trametinib inhibit the growth of c-MET/sgp53 HCC xenografts. These findings highlight the importance of biomarker-based targeted therapies for effective cancer treatment. In phase I clinical study, trametinib, and sorafenib were used to treat unselected patients with hepatocellular carcinoma. Unfortunately, the resulting clinical data indicated this combination therapy had limited efficacy in advanced HCC . HoweverTP53. Clearly, there is a need to develop human HCC cell lines with wild-type TP53, and these cell lines would be critical to validate the specificity of trametinib against TP53 null HCC cells. The c-MET signature of these cell lines has been previously analyzed [Our study has several limitations. The trametinib sensitivity data are based on three HCC cell lines  and one hepatoblastoma cell line (HepG2). They are not isogenic models. Unfortunately, there is no commercially available HCC cell line with wild-type analyzed , revealiTP53 null tumors represent an HCC subset that trametinib could specifically target. Based on the TCGA-LIHC dataset, 23 human HCC samples displayed c-MET activation and TP53 null mutation concomitantly, representing 6% of the human HCC collection. Each year, approximately 900,000 new cases of liver cancer occur worldwide [TP53 null mutation and c-MET amplification. These patients may benefit from the targeted treatment identified in this study.It is important to note that HCC is a heterogeneous disease with many subtypes. Therefore, identifying reliable biomarkers for patient selection is critical for precision HCC treatment. As revealed in our previous work , c-MET horldwide . TherefoThe plasmids used in this study, including pT3-EF1α-c-MET (human c-MET or hMET) and pCMV-sleeping beauty transposase (SB), have been described in our previous studies , 17, 34.FVB/N mice were obtained from Charles River Laboratories . To generate the c-Met/sgp53 HCC model, 20 μg pT3-EF1α-c-MET and 20/40 μg pX330-sgp53 along with 0.8 μg pCMV/SB plasmids in 2 ml saline (0.9% NaCl) were delivered to 6–8 week-old mice  by hydrodynamic tail vein injection. Mice were housed, fed, and monitored following protocols approved by the Committee for Animal Research at the University of California, San Francisco, and the University of Hawaii Cancer Center.2 for ~4 weeks. The primary liver tumor cells were generated. To obtain a stably passaged cell line, 1 × 107 primary tumor cells were injected into flanks of 6–8 week-old FVB/N mice. When the subcutaneous tumor developed, tumor tissue was dissected into small pieces and digested with trypsin . The new generation of liver tumor cells (P0) were cultured and passaged in vitro for 3 times. Then 1 × 107 “P0” cells were implanted into the flanks of a new recipient FVB/N mouse. These procedures were repeated for three rounds. The stably passaged cell line from c-MET/sgp53 HCC (MP cell) was finally generated.Mice were humanely euthanized and rinsed in 70% ethanol. The liver tumor (2–3 g) was dissected, washed in PBS, and minced into ~1 mm fragments using a scalpel blade. All procedures were operated in the hood using autoclaved dissecting tools. The tumor fragments were digested with 0.6 mg/L collagenase  at 37 °C for 15–30 min, then filtered through 100 μm nylon mesh cell strainer  and spun . The cell pellet was resuspended and cultured in 5 ml DMEM with 10% fetal bovine serum (FBS) and 1% Penicillin/Streptomycin at 37 °C and 5 % CO7 MP cells) suspension of tumor cells in Matrigel  were inoculated subcutaneously into the right flank of 6 ~ 8 week-old FVB/N mice . On day 14 post-implantation, when the tumor size reached about 100–150 mm3, mice were randomized into 5 groups for treatment. Metformin (250 mg/kg/day), niclosamide (100 mg/kg/day), cabozantinib (60 mg/kg/day), trametinib (1 mg/kg/day), or vehicle was orally administered daily via gavage. Tumor volume was measured every three days and estimated from the caliper using the formula V = A × B2/2 . In addition, body weight was measured every day. After 3 weeks of treatment, all mice were sacrificed, and tumors were harvested for further analysis.To develop mice xenografts, 200 μl . The IC50 values were calculated using the Prism 9.0 software (GraphPad Software Inc.). All experiments were repeated at least three times.The following human HCC cell lines were used in this study: LM9, HLE, HepG2, and Hep3B. They were obtained from the American Type Culture Collection . In addition, Dr. Felsher of Stanford University kindly provided the HCC3-4 and HCC4-4 cell lines, which were isolated from c-Myc mouse liver tumors. All cell lines were grown in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% FBS and penicillin/streptomycin  in a 5% COMouse liver tissues and tumor tissues were fixed in 4% paraformaldehyde, embedded in paraffin, and sectioned at 5 μm. Then, hematoxylin and eosin (H&E), and IHC staining were performed. For antigen retrieval, de-paraffinized slides were incubated in antigen retrieval buffer  and microwaved for 10 min. After a blocking step with the 5% goat serum and Avidin-Biotin blocking kit , the sections were incubated with the primary antibodies overnight at 4 °C. To quench the endogenous peroxidase, slides were then subjected to 3% hydrogen peroxide for 10 min, and subsequently, the secondary antibody was applied for 30 min at room temperature. The immunoreactivity was visualized with the Vectastain ABC Elite kit (Vector Laboratories Inc.) and DAB . Slides were counterstained with hematoxylin. The following primary antibodies were used: anti-p53 , anti-CK19 , anti-HNF4a , anti-Ki67 , anti-p-ERK , and anti-Cleaved-Caspase 3  were used in the present investigation.Tyr1234/1235 , anti-ERK1/2  and anti-p-ERK1/2Thr202/Tyr204 , anti-GAPDH , and anti-β-actin .Frozen liver tumors were homogenized in Mammalian Protein Extraction Reagent (Thermo Fisher Scientific) containing the Complete Protease Inhibitor Cocktail (Thermo Fisher Scientific). Protein concentrations were determined with the Bio-Rad Protein Assay Kit . The Supernatant was denatured by boiling in 2× Laemmli sample buffer . Equal loading was assessed by probing the membranes with the GAPDH or β-actin antibody. Aliquots of 30 μg protein lysates were separated by SDS-PAGE  and transferred onto PVDF membranes (Bio-Rad). Membranes were blocked in 10% non-fat milk in Tris-buffered saline containing 0.05% Tween-20 and incubated with primary antibodies at 4 °C overnight. Then membranes were incubated with horseradish peroxidase-secondary antibody  for 1 h at room temperature and developed with ClarityTM Western ECL Substrate . The antibodies used are as follows: anti-p53 , anti-c-MET , anti-p-METAccording to the manufacturer’s instructions, mouse genomic DNA was extracted from frozen mouse tissue samples using the Mouse Direct PCR Kit . The amplification conditions were 94 °C for 5 min, followed by 35 cycles of 94 °C for 20 s, 50 °C for 30 s, and 72 °C for 30 s. The sequences of the primers are as follows: Forward: CCTACTGGATGTCCCACCTTCT; Reverse: CAGACACCCAACACCATACCA. For individual clonal sequencing, PCR products were purified and inserted in the pGEM®-T Easy Vector  according to the manufacturer’s instructions. Next, clones were cultured, and plasmids were extracted using the Zyppy Plasmid Miniprep kit . The inserted sequence was subsequently sequenced using T7 primers.n = 3), MP cells (n = 3), and FVB/N normal livers (n = 6) using Quick-RNA Miniprep Kit . For RNA extraction, the c-MET/sgp53 HCCs were collected from male mice at 17, 20, and 22 weeks post-injection. The normal livers were collected from FVB/N male mice aged 21–28 weeks. The normal liver tissues were collected from wild-type FVB mice. The RNA quality control was determined using Agilent RNA 6000 Nano Kit  and Bioanalyzer . Library preparation and sequencing were performed by Novogene . Reads quality was controlled by fastqc (v0.11.7). Gene read counts were in Ensembl Gene ID and converted to Entrez Gene ID using Bioconductor Package Maintainer org.Mm.eg.db. Not annotated, duplicate Entrez IDs and genes without symbols were removed. R package “edgeR” were used to identify the DEGs. DEGs were limited by a p-value of 0.05 and an FDR  of 0.05. The Venn diagram was drawn using the VennDiagram package. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed using ggplot2 package. The RNAseq data for this study were deposited in the Gene Expression Omnibus database (GSE229764).Total RNA was extracted from mouse sgp53/c-MET HCCs . The overall sample size is 410, including 50 surrounding liver tissues (ST) and 360 HCC samples with TP53 mutation data. RNA sequencing data were analyzed in R using multiple packages. The analysis of c-MET activation status was performed as previously described [To investigate the relationship of c-MET activation with escribed . In brieescribed . The datt-test, Mann–Whitney test, Welch’s t-test, and Log-rank (Mantel-Cox) test analyses. The data were expressed as the mean ± SD  of at least three independent experiments.The Prism 9.0 software (GraphPad Software Inc.) was used to analyze the data. Statistical analysis was performed using Student’s Supplementary FiguresSupplementary Table S1Supplementary Table S2Supplementary Table S3Supplementary Table S4Supplementary Table S5Supplementary Table S6Original western blotschecklist


This paper studies the uniqueness of solutions to a two-term nonlinear fractional integro-differential equation with nonlocal boundary condition and variable coefficients based on the Mittag-Leffler function, Babenko’s approach, and Banach’s contractive principle. An example is also provided to illustrate the applications of our theorem. Let mentclass2pt{minimx), from .Let λ is a constant. In particular, for Let  biology –21. In 2 biology  investigμ is a constant, and In particular, for all n et al.  studied equation  by settii et al.  studied Very little is known in modern literature about boundary value problems of fractional integro-differential equations with integral boundary conditions and variable coefficients. We will work on equation , which, We define the Banach space The two-parameter Mittag-Leffler function  is definBabenko’s approach  is a powLetbe continuous functions andbe a continuous functional. Assume thatwhereMis a constant satisfyingThenis a solution to the following nonlinear integro-differential equationif and only ifsatisfies the integral equationin the spaceClearly, mentclass2pt{minimWe are now ready to present our key results regarding the uniqueness of solutions to equation  using BaAssume thatis continuous andis a continuous functional, and there exist nonnegative constantsLandsuch thatFurthermore, Then equation , we get Clearly, entclass1pt{minimaentclass1pt{minimaWe have investigated the uniqueness of solutions to the nonlinear fractional integro-differential equation  with non