Although wastewater and sewage systems are known to be significant reservoirs of antibiotic-resistant bacterial populations and periodic outbreaks of drug-resistant infection, there is little quantitative understanding of the drivers behind resistant population growth in these settings. In order to fill this gap in quantitative understanding of the development of antibiotic-resistant infections in wastewater, we have developed a mathematical model synthesizing many known drivers of antibiotic resistance in these settings to help predict the growth of resistant populations in different environmental scenarios. A number of these drivers of drug-resistant infection outbreak, including antibiotic residue concentration, antibiotic interaction, chromosomal mutation, and horizontal gene transfer, have not previously been integrated into a single computational model. We validated the outputs of the model with quantitative studies conducted on the eVOLVER continuous culture platform. Our integrated model shows that low levels of antibiotic residues present in wastewater can lead to increased development of resistant populations and that the dominant mechanism of resistance acquisition in these populations is horizontal gene transfer rather than acquisition of chromosomal mutations. Additionally, we found that synergistic antibiotics at low concentrations lead to increased resistant population growth. These findings, consistent with recent experimental and field studies, provide new quantitative knowledge on the evolution of antibiotic-resistant bacterial reservoirs, and the model developed herein can be adapted for use as a prediction tool in public health policy making, particularly in low-income settings where water sanitation issues remain widespread and disease outbreaks continue to undermine public health efforts.IMPORTANCE The rate at which antimicrobial resistance (AMR) has developed and spread throughout the world has increased in recent years, and according to the Review on Antimicrobial Resistance in 2014, it is suggested that the current rate will lead to AMR-related deaths of several million people by 2050 . One major reservoir of resistant bacterial populations that has been linked to outbreaks of drug-resistant bacterial infections but is not well understood is in wastewater settings, where antibiotic pollution is often present. Using ordinary differential equations incorporating several known drivers of resistance in wastewater, we find that interactions between antibiotic residues and horizontal gene transfer significantly affect the growth of resistant bacterial reservoirs. Salmonella enterica serotype Typhi infections spread around sewage lines in the city of Hyderabad in Pakistan  and no plasmid-containing bacteria such that there was only chromosomal mutation as a mechanism for resistance. This experimental validation was done on the eVOLVER, a continuous culture platform which allows for automated, highly flexible and scalable microbial growth and lab evolution. eVOLVER allows for continuous flow conditions and independent, precise, and multiparameter control of growth conditions such as temperature and flow rate  (S1). This was added to match the experimental finding that resistance can coexist at a steady state with the susceptible population. We theorized that the observed susceptible population also contained a subpopulation of semi-resistant cells which were not detected by the 8× MIC resistance cutoff rate of the experiment. These semiresistant cells would allow for a sustained susceptible population in conditions with constant antibiotic pressure.In order to validate the developed model, experiments were conducted with low rate . This alin vitro behavior allowing for the development of a model with predictive capabilities.After adjustments, the resulting model outputs matched the general behavior of the resistant and susceptible bacterial populations in response to the selected concentrations of rifampicin  to f. ThFollowing this initial study, the adjusted model was used to make predictions for a second experiment with different rifampicin concentrations  and b. TE. coli under low antibiotic pressure. In the scenario of no HGT, as expected, only mutational resistance was observed . This is in agreement with several prior studies linking subtherapeutic antibiotic levels with both chromosomal resistance mutation development and HGT  was observed to decrease the time it takes for the resistant population to dominate under subinhibitory concentrations. This result is in agreement with previous studies showing that increased selective pressure from antibiotics at subinhibitory concentrations can increase resistance development  over time as well as equations modeling the susceptible population (S) and populations resistant to both antibiotics 1 and 2 from chromosomal mutation (mR) or from HGT, specifically a resistance-conferring plasmid (pR) over time (R1 and R2). Each antibiotic is modeled with terms for the antibiotic residue concentration in the environment (E) and the antibiotic clearance rate (ek). Each bacterial population is modeled with terms for growth rate (α), which is limited by the carrying capacity (Nmax). The bactericidal activities of the antibiotics are modeled by the term for killing rate in response to each antibiotic . These killing terms are modified by the antibiotic concentration where the killing action is half its maximum value for either susceptibility syn) with syn < 1 indicating antagonistic interaction and syn > 1 indicating synergistic interaction. Also included in the model are terms for bacterial inflow  and outflow (Tk) rates (m) which are concentration dependent. Parameters governing mutation rate and antibiotic interaction are based on a previously developed model of Michel et al. (β). HGT model terms and parameter values are adapted from a model of HGT in agricultural waste of Baker et al.  rates . These il et al. . HGT is r et al. . Other m studies , 29. We E. coli MG1655 in static conditions at 37°C. Then, inflow and outflow of rifampicin-containing LB medium at three concentrations  were started at a flow rate of 16 ml/h. Following the beginning of inflow/outflow, each culture condition was sampled daily, and the concentration of total bacteria was monitored through optical density (OD) measurements taken through the eVOLVER system and resistant bacteria were calculated through plating on selective LB agar containing 200 mg/liter rifampicin. This rifampicin concentration is eightfold greater than the experimentally determined MIC for rifampicin (25 mg/liter). After the conclusion of the experiment, model equations and parameter values were adjusted to match the experimental behavior . The experiment was initialized with inoculation of LB medium with behavior .


Background: Inappropriate antibiotic use among outpatients is recognized as the primary driver of antibiotic resistance. A proper understanding of appropriate antibiotic usage and associated factors helps to determine and limit inappropriateness. We aimed to identify the rate of appropriate use of antibiotics and identify factors associated with the inappropriate prescriptions. Methods: We conducted a cross-sectional descriptive study in outpatient antibiotic use at a hospital in Can Tho City, Vietnam, from August 1, 2019, to January 31, 2020. Data were extracted from all outpatient prescriptions at the Medical Examination Department and analyzed by SPSS 18 and Chi-squared tests, with 95% confidence intervals. The rationale for antibiotic use was evaluated through antibiotic selection, dose, dosing frequency, dosing time, interactions between antibiotics and other drugs, and general appropriate usage. Results: A total of 420 prescriptions were 51.7% for females, 61.7% with health insurance, and 44.0% for patients with one comorbid condition. The general appropriate antibiotic usage rate was 86.7%. Prescriptions showed that 11.0% and 9.5% had a higher dosing frequency and dose than recommended, respectively; 10.2% had an inappropriate dosing time; 3.1% had drug interactions; and only 1.7% had been prescribed inappropriate antibiotics. The risk of inappropriate antibiotic use increased in patients with comorbidities and antibiotic treatment lasting >7 days (p < 0.05). Conclusions: The study indicated a need for more consideration when prescribing antibiotics to patients with comorbidities or using more than 7 days of treatment. Klebsiella pneumoniae infections due to resistance in some countries [Staphylococcus aureus observed were methicillin-resistant and Escherichia coli resistant to third generation cephalosporins was detected with 36.0% in 2019 [Ever since antibiotics were introduced into medicine in the 1940s, resistance to them has also been ascertained and acknowledged [ountries . The WHO in 2019 . In 2013 in 2019 ,8.In the United States, studies have described the inappropriate selection of antibiotics and the need for rational antibiotic prescribing practices. Among outpatient antibiotic prescriptions filled, 23.2–40% were inappropriate ,10,11, aMany factors that may be associated with inappropriate antibiotic use by outpatients can be related to characteristics of prescribing physicians, patients, the environment, or the performance of physical examinations ,15. In VThe complicated situation of antibiotic resistance has, in recent years, raised great concern among local health facilities. Being a hospital with well-trained healthcare staff and associated with teaching/training activities, the Hospital of Can Tho University of Medicine and Pharmacy provides healthcare services for Southwestern Vietnam, with a large number of patients per day and a variety of clinics. The hospital is consistently attentive in the safe, effective, and appropriate use of drugs, especially antibiotics. Several measures to promote the appropriate use of antibiotics have been surveyed and proposed in the hospital, but these focus mainly on inpatient and prophylactic antibiotics. In this study, for a total evaluation of antibiotic use in outpatient treatment, we identified the rate of the appropriate use of antibiotics and assessed a number of factors associated with inappropriate prescription in this setting.We conducted a cross-sectional descriptive study to analyze outpatient antibiotic prescriptions in clinics of a hospital in Can Tho City, Vietnam. In the region, this general hospital serves a large number of patients from different regions, with a total of 205,848 outpatients and 12,074 inpatients in the year 2019.We included outpatient prescriptions with antibiotics and excluded prescriptions (1) with no oral antibiotics  and (2) lacking information on the indicated drug. Prescriptions repeated at the re-visits of the sampled patients were also excluded.Data were extracted from all outpatient prescriptions recorded in the hospital’s prescription database for 6 months  at the Medical Examination Department of the Hospital of Can Tho University of Medicine and Pharmacy. Screening of subjects before collection was based on inclusion and exclusion criteria. The eligible prescriptions from a total of 97,126 outpatient prescriptions during the study period were listed in numerical orders; subsequently, the prescriptions numbered at a defined interval were collected according to the technique of systematic random sampling, until 420 prescriptions were reached. Baseline data included demographic characteristics, health insurance, comorbidities, and characteristics of antibiotic use. Prescriptions lacking information on the prescribed drugs were excluded.The sample size was estimated by using the formula for a single proportion with an estimated proportion of antibiotic appropriate use which was 0.847, as reported in a prior study , an assuwww.drugs.com; accessed on 2 March 2020) to look up interactions between antibiotics with other drugs prescribed and considered interactions labeled “Major” by entering the drug list in the prescriptions into the tool and checking for the interactions. A prescription was generally considered appropriate when all five of the indicators listed above were appropriate. This study also assessed the effects of several relevant factors, including age , gender, health insurance, comorbidities, and duration of antibiotic treatment (≤7 days and >7 days) on appropriate antibiotic use.Assessment of the appropriateness of the antibiotic use was according to the following documents, in this order of priority: (1) medication instruction sheet, which was revised and approved by the regulatory authority—Drug Administration of Vietnam; (2) Instructions for Use of Antibiotics of Vietnam’s Ministry of Health, issued with Decision No. 708/QD-BYT, dated 2 March 2015; and (3) Vietnamese National Drug Formulary 2018, which specified the use of each drug substance [p < 0.05 level of statistical significance.Collected data were analyzed by using SPSS 18. Descriptive statistics, such as frequencies and percentages, were included in the analysis. The frequencies of categorical variables of two groups were compared by using the Chi-squared test, with 95% confidence intervals and a p < 0.05 was considered statistically significant.We modeled a multivariable logistic regression analysis, assuming the inappropriate usage as the outcome variable and age group, gender, health insurance, comorbidities, and duration of antibiotic treatment as explanatory ones. All five explanatory variables were analyzed in both models of univariable and multivariable analysis. A From a total of 97,126 prescriptions, after screening based on inclusion and exclusion criteria, 852 antibiotic prescriptions were eligible for systematic random sampling and 420 prescriptions were included for this study. Of 420 prescriptions, those for female patients were 51.7%. The mean age was 40.6 years (SD: 18.4). Antibiotics prescribed for outpatient treatment accounted for the highest proportion for the age group 16–59 (77.9%) and lowest for the age group ≥ 60 years (20.2%). In total, 61.7% had health insurance. The largest percentage of patients had one comorbid condition (44.0%), followed by those with no comorbidities (41.0%); the lowest percentage had two comorbidities (4.3%). The prescriptions collected from the ear–nose–throat clinic accounted for the highest proportion (35.7%), and those from the oncology clinic for the lowest (0.5%) .Of 420 prescriptions, the general appropriate rate of antibiotic use accounted for 86.7%. Of these, 98.3% of the prescribed antibiotics were considered appropriate. Higher-than-recommended antibiotic dose and frequency were found in 9.5% and 11.0%, respectively. Dosing time without adherence to medical recommendations accounted for 10.2%. The rate of antibiotic interactions with other drugs was 3.1% . Penicilp > 0.05). The risk of inappropriate antibiotic use increased in patients with comorbidities  and duration of antibiotic treatment > 7 days , with a statistically significant difference.Results of the univariable analysis reported no association between age, gender, or health insurance and inappropriate antibiotic usage (p < 0.05)  .Antibiotic prescribing in the children and elderly age group was lower than reported rates in a study at in public health facilities in Hoa Nhon district, Binh Dinh province with 46.12% of antibiotic prescriptions was performed on age group ≤15 years and 22.15% belonged to age group ≥60 years . In genen = 45, 63.4%) [Assessing whether or not the use of antibiotics was appropriate can be related to many factors. In this study, the appropriate usage of antibiotics was evaluated according to whether the prescribed antibiotic followed the recommendation indication or not. We recorded 413 (98.3%) of 420 prescriptions involving antibiotic selection in accordance with recommendations, and none containing antibiotics involving absolute contraindication for patients. In addition, no prescription had more than one antibiotic containing the same active ingredient. The selection of appropriate antibiotics in the current study did not differ from that of the previous study in different hospitals, and all were found to be more than 93% appropriate . Neverth, 63.4%) . These rThe second factor assessed was the dosage of antibiotics for the successful treatment of infections and reduction in resistance rate. We found that 90.5% of prescriptions had an appropriate dosage; 9.5% prescribed a higher-than-recommended dose, and no one had a lower-than-recommended dose. Vo Thi Thanh Thuy recorded 98% of cases using antibiotics with the appropriate dosage, but 2% with doses lower than recommended . In prevThird, the selection of dosing frequency was also a significant issue. In some cases, a single dose was appropriate, but due to a higher or lower frequency than recommended, resulting in a 24-h dose not being suitable. In our study, the rate of appropriate dosing frequency was quite high (89.0%); 11.0% of cases were higher than recommended. In addition, in 89.8% of prescriptions, we found the timing of antibiotic use to be correct. According to research at the central general hospital, errors in the time for using antibiotics were the most common error, at a rate of 32.6% . The difThe fifth factor, drug–drug interaction, is an important problem related to simultaneous use of multiple drugs. In our research, of 420 prescriptions examined, the interaction rate between antibiotics and other drugs was low, at 3.1%. However, only major interactions that are highly clinically significant and avoid combinations were recorded in this study. On the other hand, fewer interactions between antibiotics with the other drugs were acknowledged in comparison with those of cardiovascular drugs in general. In a study at Hue University of Medicine and Pharmacy Hospital, the percentage of prescriptions appearing to have clinically significant interactions was higher than our results, with 6.7%. Furthermore, the reported interaction rate was relatively higher in a study at Cho Ray hospital, Ho Chi Minh City, Vietnam, with 18.7% . Due to In the viewpoint of the quantity of antibiotic use, Vietnam ranks 11th in antimicrobial consumption per capita, with 32 Defined Daily Doses per 1000 inhabitants per day and is the second low- and lower-middle income country to appear on the list of global antibiotic consumption by country between 2010 and 2015 . ConsideAdministration of antibiotics to a patient involves many objective factors, ranging from the patient’s characteristics to the disease condition. Normally, physicians choose antibiotics for therapy depending on two factors: the characteristics of the patient and the pathogenic bacteria. The patient’s characteristics should be scrutinized, such as age, drug-allergy history, liver and kidney function, immunodeficiency status, and comorbidities. Regarding pathogenic bacteria, the type of bacteria causing the disease and their sensitivity to antibiotics should also be considered in the treatment of infections. Decision-making in antibiotic prescribing among doctors in Vietnam is influenced by both disease-related characteristics and individual factors, including acceptable minimum treatment coverage . Howeverp < 0.05.Using univariate analyses and multivariate logistic regression analysis, we found the association between comorbidities, duration of antibiotic treatment, and inappropriate antibiotic usage to be p = 0.009. In a series of nine studies evaluating comorbidities as a prescribing factor, seven found no association, and only two studies demonstrated an association between the presence of comorbid symptoms and the prescription [The rate of using antibiotics unreasonably with patients with comorbidities was 16.9%, 2.30 times higher than in patients without comorbidities (8.1%). This difference was statistically significant, with OR = 2.30 (1.21–4.36) and cription . For patp = 0.002. The duration of antibiotic therapy depended on the purpose for use  and the patient’s course [Furthermore, we found that, in outpatients using more than 7 days of antibiotic treatment, the rate of using antibiotics inappropriately was 28.6%; this was 3.04 times higher than the group of patients administered antibiotics within 7 days (11.6%). The difference was statistically significant, with OR  = 3.04 (1.45–6.36) and s course . In outps course . This coRegarding age and gender, according to the results of univariate analyses and multivariate logistic regression analysis, we had yet to find an association of these two factors with inappropriate use of antibiotics for outpatients. In fact, the prescription of an antibiotic would depend on the characteristics of each age group. According to Rachel McKay et al. (2016), after they retrieved a total of 2848 abstracts and analyzed 97 included in full-text reviews and 28 meeting full inclusion criteria, the result showed that just one of ten studies exploring gender found a statistically significant association between being male and having higher odds of antibiotic prescription . Among nRegarding health insurance, this was the most important policy in the Vietnamese social security system, ensuring that all citizens had health care, not for profit, and implemented by the Vietnamese government. In order to limit compensations, the Ministry of Health of Vietnam and the hospitals had proposed treatment regimens for each disease, creating a scientific basis for appropriate drug indications. In addition, hospitals had applied information technology to medical examinations and treatment to assist physicians in prescribing. Specifically, on the hospital management software, all drugs were registered with the number of uses per day, route and dose, thereby providing guidance to support physicians in prescribing according to the correct treatment regimen, whether with or without health insurance . UltimatOverall, comorbidities and antibiotic use longer than 7 days made the risk of inappropriate antibiotic use among outpatients respectively 2.46 and 2.63 times higher.For the fight against antimicrobial resistance (AMR) global burden, a crucial health approach providing educational programs for all doctors, especially young doctors, was supported in a study in Italy. As suggested by the study, a specific course and training on antibiotics should be included in the core curriculum of the School of Medicine; meanwhile, an antimicrobial-resistance program should be set up in health districts and hospitals; and the institution of a network on AMR, with the AMR sentinel doctors directly involved in monitoring and evaluating trends in AMR in their health districts and hospital should be set up . FurtherThe rate of appropriate general antibiotic usage was 86.7%. Patients with comorbidities and antibiotic treatment lasting >7 days had inappropriate rates of using antibiotics respectively higher than 2.46 and 2.63 times, as compared to the other group of patients. Therefore, physicians need to consider more carefully when prescribing antibiotics to patients with comorbidities or when administering antibiotics with a treatment longer than seven days. Further research is needed to examine intervention solutions focusing on antibiotic use in outpatients to ensure safe, effective, and economical use of antibiotics, and to reduce antibiotic resistance. Moreover, the initial steps for promoting training on AMR and implementing AMS in Vietnam should be taken into further consideration.


Currently, assays for rapid therapeutic drug monitoring (TDM) of β-lactam antibiotics in blood, which might be of benefit in optimizing doses for treatment of critically ill patients, remain challenging. Previously, we developed an assay for determining the penicillin-class antibiotics in blood using a thermometric penicillinase biosensor. The assay eliminates sample pretreatment, which makes it possible to perform semicontinuous penicillin determinations in blood. However, penicillinase has a narrow substrate specificity, which makes it unsuitable for detecting other classes of β-lactam antibiotics, such as cephalosporins and carbapenems. In order to assay these classes of clinically useful antibiotics, a novel biosensor was developed using New Delhi metallo-β-lactamase-1 (NDM-1) as the biological recognition layer. NDM-1 has a broad specificity range and is capable of hydrolyzing all classes of β-lactam antibiotics in high efficacy with the exception of monobactams. In this study, we demonstrated that the NDM-1 biosensor was able to quantify multiple classes of β-lactam antibiotics in blood plasma at concentrations ranging from 6.25 mg/L or 12.5 mg/L to 200 mg/L, which covered the therapeutic concentration windows of the tested antibiotics used to treat critically ill patients. The detection of ceftazidime and meropenem was not affected by the presence of the β-lactamase inhibitors avibactam and vaborbactam, respectively. Furthermore, both free and protein-bound β-lactams present in the antibiotic-spiked plasma samples were detected by the NDM-1 biosensor. These results indicated that the NDM-1 biosensor is a promising technique for rapid TDM of total β-lactam antibiotics present in the blood of critically ill patients. The optimization of antibiotic dosing regimens for patients with antimicrobial-resistant (AMR) infections has the potential to dramatically improve therapeutic outcomes while reducing the development of antibiotic resistance. This is especially important when treating critically ill patients where administration of standard antibiotic dosages sometimes results in subtherapeutic antibiotic blood concentration levels that adversely affect treatment outcomes ,2. This Currently, β-lactam antibiotics are the most widely used group of antimicrobial agents due to their safety, efficacy and broad spectrum . The β-lUnlike routine monitoring of aminoglycoside and glycopeptide antibiotics using commercial assays, the methods used to monitor β-lactam antibiotics have been largely developed in-house . Among tThe Enzyme Thermistor is a flow-injected biosensor device that combines an immobilized enzyme for recognition and catalysis of the substrate with ultrasensitive thermometric detection. The temperature changes generated by the catalytic enthalpy of the enzymes are used to quantify the presence of specific substrates, such as antibiotics ,22. NumeIn order to meet clinical requirements for determination of most therapeutic β-lactam antibiotics, a broad analytical capacity for the most applied antibiotics, in particular cephalosporins and carbapenems, was verified by comparison between the NDM-1 biosensor and the penicillinase biosensor.Performance in the activity measurement of two cephalosporins (ceftriaxone and cefepime) and two carbapenems (meropenem and imipenem) was assessed and compared using the antibiotics spiked in HEPES buffer while using penicillin G and aztreonam as two references. It was observed that with the antibiotic concentration at 300 mg/L, the NDM-1 biosensor showed significantly greater activity responses to all tested cephalosporins and carbapenems than the penicillinase biosensor except for two comparable activity responses to penicillin G and aztreonam . The resBlood measurement of the β-lactam antibiotics spiked in plasma was studied under two different blood sampling conditions, a heparin sodium tube and an EDTA tube. Clinically, plasma samples are usually prepared from whole blood collected by tubes with heparin sodium or EDTA as the anticlog agent. For comparison, the effect of these tubes on the measurements of blood antibiotics is shown in The properties of the NDM-1 biosensor in the determination of carbapenems (imipenem and meropenem); the second, third and fourth generations of cephalosporins ; and cephamycins (cefoxitin) in blood plasma were evaluated with the antibiotics spiked in HEPES buffer as control. The real responses of the NDM-1 biosensor to cefoxitin are represented in Nowadays, some FDA-approved β-lactam antibiotics are coformulated with β-lactamase inhibitors aiming to improve therapeutic efficacy. For instance, piperacillin is coformulated with tazobactam with a mass ratio of 4:1, ceftazidime is coformulated with avibactam with a mass ratio of 4:1 and meropenem is coformulated with vaborbactam with a mass ratio of 1:1 . To evalMultiple lines of evidence support that the combination of one β-lactam antibiotic with another antibiotic exhibits synergism in antimicrobial activity in vitro . MoreoveIn the blood of patients treated with an antibiotic, the antibiotic exists in protein-bound and unbound  forms, and the proportion of the antibiotic binding on protein depends on the intrinsic property of the antibiotic. In order to identify which antibiotic form the NDM-1 biosensor responded to in the spiked plasma, a study was performed using the NDM-1 biosensor to detect and compare the activity responses to the filtrate and the concentration generated from ultrafiltration of meropenem- or penicillin G-spiked plasma. After ultrafiltration, the protein-bound and unbound forms of the antibiotic are concentrated in the concentration and the filtrate, respectively. The meropenem and penicillin G were reported as having a low activity (2%) and a high activity (65%) to bind plasma protein, respectively . As showClinically, sufficient PK data points are required for TDM of β-lactam antibiotics for the purpose of optimizing dosage . TherefoIn order to cover a broad range of antibiotics applied in therapeutics, a versatile β-lactamase that is capable of hydrolysis of most classes of β-lactam antibiotics was employed in the thermometric biosensor assay. As compared with our previous penicillinase-based biosensor assay for detection of limited types of the penicillin-class antibiotics in blood , NDM-1 wf T>MIC) determines microbicidal efficacy [f T>MIC for the given pathogen [f T>4–5 × MIC) were chosen for patients with severe infection or continuous renal replacement therapy [Cmax) of almost all β-lactam antibiotics in plasma are below 200 mg/L after antibiotic administration, 200 mg/L was set up as the upper concentration of the calibration curve of the tested antibiotics in this study. Since the lower limits of quantification (LLOQs) of the NDM-1 biosensor were 6.25 mg/L for detecting cefaclor, cefoxitin, meropenem and imipenem and 12.5 mg/L for detecting ceftazidime and cefepime in the spiked plasma  antibiotic concentration is greater than the minimal inhibitory concentration (MIC) , which are classified as the β-lactam inhibitors such as sulbactam and tazobactam, the diazabicyclooctane (DBO) inhibitors such as avibactam  and the Meropenem (MEM) and ceftazidime (CAZ) were obtained from USP . Penicillin G (PEN), cefoxitin (FOX), cefaclor (CEC), ceftriaxone (CTRX), cefepime (FEP) and tazobactam (TAZ) were from Solarbio . Avibactam (AVI), vaborbactam, fosfomycin (FOS), amikacin (AMK) and imipenem (IMI) were purchased from MedChemExpress . The purity of all antibiotics used in this study was higher than 95%. The propylamino-derivatized controlled pore glass (CPG) beads, with a diameter of 125–140 µm and pore size of 50 nm, were obtained from Steinachglas . HEPES was purchased from Sangon Biotech . All other chemical reagents were purchased from Sigma-Aldrich  unless stated otherwise. Molecular biology enzymes were bought from Takara Bio , and the primers were purchased from Sangon Biotech .2+. The unit activity of rNDM-1 to hydrolyze meropenem was 108 IU/mg.The coding sequence of the full-length NDM-1 gene (GenBank Accession No. JN616388) was synthesized . Subsequently, 28 amino acids were removed from the N-terminus and an N-terminal His-6-tag was added to the NDM-1 gene using PCR, and the product was inserted into the expression vector pET-28a (Novagen). The expression and purification of recombinant NDM-1(rNDM-1) were performed as previously described except that the His-6 tag was not removed from the N-terminus of the recombinant enzyme . The purThe NDM-1 biosensor consisted of an immobilized NDM-1 enzyme column, a peristaltic pump  connecting with an injection valve with a 350 µL sample volume and the Enzyme Thermistor instrument . The working temperature was set to 30 °C, and the operational flow rate was 10.The NDM-1 column was loaded with 400 µL CPG beads with immobilized rNDM-1 enzyme. The NDM-1-immobilized CPG was prepared using 400 µL CPG beads and 100 units of purified rNDM-1, and the remaining active sites were blocked with 1% BSA prepared with 0.01M PBS for the purpose of reducing background signal from plasma metrics. The penicillinase column was prepared as previously described .β-Lactam antibiotics used in this study were dissolved with deionized (18.2 MΩ) water to prepare antibiotic stock that was further diluted to the working solutions with 50 mM/L HEPES  at pH 7.5. The HEPES buffer was also used as the running buffer for the biosensor system throughout the study.Blood samples were collected from healthy volunteers using either heparin sodium tubes or EDTA tubes. Within two hours after collection of the blood, plasma samples were obtained by centrifuge and immediately stored in a −80 °C freezer until experiment.In the biosensor determination of antibiotic samples, the antibiotic stocks were serially diluted with the HEPES buffer to the defined concentrations and a volume of 450 µL for each sample to be tested. For determination of antibiotic-spiked plasma samples, the antibiotic stocks were firstly serially diluted with the HEPES buffer, followed by mixing with plasma to a series of defined concentrations of the testing antibiotic samples, including a HEPES buffer-spiked plasma sample with blank antibiotic as a negative control. The net value of the measured antibiotic concentration was recalibrated by deduction of the control signal from the total measurement signal.g to obtain the filtrate fraction in the lower chamber and the concentrate fraction in the upper chamber. To determine antibiotic distribution in the filtrate fraction and the concentrate fraction, the filtrate fraction was directly analyzed by the NDM-1 biosensor, and the concentrate fraction was reconstituted (diluted) with the running buffer to 450 μL before analysis by the NDM-1 biosensor. The net activity response (Δmv) of the NDM-1 biosensor to the spiked antibiotic in the concentrate fraction was calculated as (the activity response to the reconstituted concentration fraction from the antibiotic-spiked plasma — the activity response of the reconstituted concentration fraction from the buffered plasma) × dilution factor.Four hundred fifty microliters of plasma samples spiked with antibiotics or buffer was loaded into the upper chamber of an Amicon Ultra-0.5 mL 30 KD cut-off centrifugal filter device  for centrifugation for 10 min at 14,000× t-test was used to compare differences throughout the study. All tests were considered statistically significant at p < 0.05.Statistical analysis was performed with Prism 8.0 . Unpaired Student’s In this study, we have developed a novel thermometric NDM-1 biosensing assay for rapid and quantitative measurements of total cephalosporin and carbapenem β-lactam antibiotics in blood without the need for any pretreatment of samples in order to reduce the measurement cycle period to within 2–3 h. A broad detection range to cover therapeutic windows of most β-lactam antibiotics for the treatment of critically ill patients was achieved using the NDM-1 as catalysis for thermometric biosensor, which allows for a routine TDM of β-lactam antibiotics in the clinical setting for critically ill patients. In addition, a potential application in TDM of original cephalosporins and carbapenems, as well as BLIs coformulated with β-lactam antibiotics, in blood plasma was evaluated. The data demonstrated that the NDM-1 biosensor is suitable for the rapid detection of plasma totals of multiple classes of β-lactam antibiotics with high sensitivity, speed and efficiency.


In this paper, properties of concretes incorporating recycling waste and corrosion susceptibility of reinforcing steel bars were studied. It was established that fineness of ground granulated blast furnace slag (GGBFS) and fly ash (FA) and their simultaneous combination have an influence on the kinetics of strength development of Portland cements and concretes. The compressive strength of concrete containing 10% by mass of GGBFS and 10% by mass of FA even exceeds the compressive strength of control concrete by 6.5% and concrete containing 20% by mass of GGBFS by 8.8% after 56 days of hardening. The formation of the extra amount of ettringite, calcium hydrosilicates as well as hydroaluminosilicates causes tightening of a cement matrix of concrete, reducing its water absorption, and improving its resistance to freezing and thawing damage. Environmental problems are becoming more and more serious from year to year. Despite the remarkable development of scientific and technological progress, some industrial processes result in the significant production of waste, which cannot be landfilled completely and present a big problem for both the producer and the environment . The wasThe cementing materials and products on their basis have played a very important role in the development of construction for many years. The Portland cements and concretes are the most used materials in the construction industry ,3. The tGGBFS is rather valuable waste and can be recycled in environment processes, agriculture, and the construction industry. Ground granulated blast furnace slag has begun to be used widely since the discovery of its latent hydraulic properties. Many researchers have been conducting lots of experiments to determine the properties of cements and concretes containing GGBFS and fly ash ,19,20,21FA is a by-product of electricity generating plants. It is used both in the technology of cement and concrete production ,27. It hSanytsky et al.  and Li eFernández et al.  also poiThe use of GGBFS and FA may also have an influence on the initial pH of the cementitious system and the protective passive layer, which is formed on the reinforcing steel bars due to the alkaline environment in concrete . In the According to Yeau and Kim , the corThis is why the aim of this research was to establish the effect of fineness of GGBFS and unactivated fly ash on the strength of Portland cements and properties of concretes as well as the level of Portland cement replacement without the reduction of compressive strength in comparison with the control cement and concrete without SCMs and to study the behavior of reinforcing steel bars in concretes incorporating such recycling waste.2/kg, respectively. The commercially available lignosulfonate based plasticizer  with a specific gravity of 1.18 and solid content of 35% was used in the research. The properties of Portland cement are presented in Portland cement CEM I 42.5 , ground granulated blast-furnace slag , and unactivated fly ash (Class F) derived from the Burshtyn power plant  were used in this study as supplementary cementitious materials. The specific surfaces of GGBFS and unactivated FA were 310, 380, 510 and 310, 510 mThe Portland cements of the second type CEM II, especially CEM II/A-S, is very popular in the construction market. The content of additions can range between 6–20% by mass in this cement according to EN 197-1. This is why the maximum amount of Portland cement (20% by mass) was replaced with GGBFS in the first stage of the research.The properties of Portland cements and aggregates were determined according to Ukrainian standards ,45,46,47The specific surface of Portland cement, GGBFS and FA was determined using the Tovarov device . The Vicat apparatus  was used to determine the setting time. The specific surfaces of GGBFS and fly ashes and residues in sieve 008 (the mesh opening size was 0.08 mm) are shown in The chemical composition of Portland cement, ground granulated blast-furnace slag and fly ash, which was determined by x-ray spectrometer ARL 9800 XP , is shown in Fly ash in comparison with Portland cement and GGBFS is characterized by a higher amount of silica, alumina, and iron oxides, which provide its pozzolanic activity.The morphology and chemical composition of the main mineral components of the studied materials were determined using scanning electron microscope SEM FEI Quanta 250 FEG, equipped with EDS .3 were molded according to 51], respTwelve specimens (six basic and six control specimens) of each concrete composition were molded and cured 28 days according to the Ukrainian standard (at the temperature of 20 ± 3 °C and the relative humidity of 95 ± 5 °C) to determine the freeze–thaw resistance of concretes. Then, the accelerated method was used according to  to deter3, S = 800 kg/m3, G (5–10 mm) = 1150 kg/m3, W = 175 kg/m3, plasticizer . The consistency class of fine-grained concrete mix was S1. Twenty percent by mass of Portland cement was replaced with 10% by mass of GGBFS and 10% by mass of FA in a reference test fine-grained concrete. Nine similar specimens (prisms of 40 × 40 × 160 mm3) of each mixture were molded. A smooth steel bar (working electrode) with the diameter of 6 mm and length of 120 mm was placed in the center of the mold and secured to prevent movement during filling and compaction. Before making the specimens, steel bars were cleaned using emery paper and then degreased with acetone. All specimens were cured 28 days at the temperature of 20 ± 3 °C and relative humidity of 95 ± 5 °C. Three specimens were tested after 28 days of hardening in the above-mentioned conditions. The other six specimens of each mix-proportion were kept in conditions of daily alternating wetting and drying . Three specimens were tested after three and six months. Before measurement, specimens were saturated with drinking water by boiling them for 3 h. Before the beginning of the test, concrete was chipped from one end of the beam specimen, exposing the reinforcing bar by 20 mm ± 10 mm. Then the working, reference, and counter electrodes were connected to the polarization circuit. Measurements of the magnitude of the current in microamperes were carried out in concrete specimens in (60 ± 5) min after turning on the potentiostat. The output of the potentiostat was raised, and the current recorded when passing the range from the steady-state potential to plus 1000 mV for 60 min. For each test specimen, the surface area of the working electrode, which was in direct contact with the concrete, was calculated. Using the calculated area of the rebar, the current density was determined.The effect of Portland cement replacement with 10% by mass of GGBFS and 10% by mass of FA on the corrosion susceptibility of reinforcing steel in concrete was studied by the potentiostatic electro-chemical test according to . ConcretThe strength of building materials is an important characteristic, which defines their quality. This study focused mainly on the influence of SCMs on the compressive strength of Portland cements and properties of concretes as well as the corrosion susceptibility of reinforcing steel bars. It is a well-known fact that there are two schemes that show how GGBFS can be added to Portland cement. It can be mixed with constituents of cement by intergrinding or after previous separate grinding to the appropriate specific surface. The second way was used as it is more effective. It allows for the achievement of a more complete realization of potential hydraulic properties of GGBFS, because the size of the slag particles remained rather coarse after intergrinding of clinker and GGBFS due to the poor grindability of the GGBFS. Therefore, the Portland cement and GGBFS, ground to the appropriate specific surface previously, were mixed in the proportions that are shown in 2/kg was 34, 21 and 16% lower than the control mortar (C) after 2, 7, and 28 days of hardening, respectively. The increase of fineness of GGBFS caused gradual growth of strength at both early and later ages in comparison with CS310. The compressive strength of mortar CS380 was reduced by 6% after 28 days of hardening, while CS500 almost reached the compressive strength of the control mortar. However, GGBFS with a specific surface of 380 m2/kg was used for further research, because the process of slag grinding up to 500 m2/kg would be a rather expensive and simultaneously time consuming process. Moreover, GGBRS is becoming more and more expensive on the market year by year, and scientists are looking for SCMs to replace it.As can be seen from 2/kg and 510 m2/kg was used in the research. The Portland cement CEM I 42.5 was replaced with 5% (CF5(310), CF5(510)), 10% (CF10(310), CF10(510)) and 15% (CF15(310), CF15(510)) by mass of FA. The obtained results are presented in Widespread by-products such as fly ash, which needs to be utilized, were used to improve the properties of cements. The specific surface of fly ash can be different and range widely at the thermal power plant. The unactivated fly ash with specific surfaces of 310 m2/kg was rather coarse and 5% by mass of Portland cement could only be replaced without the compressive strength reduction. At the same time, the usage of fly ash with a higher specific surface (510 m2/kg) made it possible to increase the replacement level of Portland cement without the reduction of strength after 28 days. The results in 2/kg was used, but the replacement level could be increased to 10% by mass.A reduction in compressive strength was observed in mortars CF10(310) and CF15(310). Nicoara et al.  observed2/kg (CS10(380)F10(510)) was studied. The mortar incorporating 20% by mass of GGBFS (CS20(380)) was used for comparison. The above-mentioned mortars containing the same amount of SCMs (20% by mass) was studied and compared with control mortar C. The results in The literature review and analysis of the obtained results show the necessity of combining GGBFS and FA. Compressive strength of mortars containing 10 and 15% by mass of GGBFS as well as 5 and 10% by mass of FA with specific surfaces of 310 (CS15(380)F5(310)) and 510 mHowever, the kinetics of the strength gain of mortars in which Portland cement was replaced with both GGBFS and fly ash was lower for up to two days, but due to the synergetic effect of the combination of these two by-products, accelerated, leveled CS10(380)F10(510), and even exceeded CS15(380)F5(310) in the compressive strength of control mortar at later ages (28 days). The existence of this effect was also confirmed by many researchers ,34,55.2O3 and SiO2. The above-mentioned hydration products appear in the vacant space of cement matrix and also on the fly ash surface F10(510) to freezing and thawing damage is obtained due to the improvement of the pore structure. The higher amount of secondary CSH gel obtained due to the pozzolanic reaction results in a decrease in the capillary porosity of concrete incorporating GGBFS and FA, which confirms the efficiency of the simultaneous combination of these SCMs.The compressive strength reduction of designed concretes is reported in 2 at 1 V for RCC and RTC, respectively. The content of calcium hydroxide in RCC was lower because 20% by mass of Portland cement was replaced with SCMs and the pozzolanic reaction also consumed some amount of Ca(OH)2, reducing its content in hardened concrete, and likely a thinner protective passive layer formed on the rebars. Over time, the current density deference of compared concretes was reduced to 7.1 µA/cm2 after 90 days and even almost levels after 180 days at a potential of 1 V due to the formation of the tighter structure of concretes incorporating GGBFS and FA. Polder [The replacement of cement with SCMs in concretes also results in the reduction of the clinker and as a result, calcium hydroxide, which is produced during Portland cement hydration and is responsible for creating a protective passive layer on the rebars due to the alkaline environment. This is why the corrosion susceptibility of reinforcing steel bars in such concretes was studied. As can be seen from . Polder  also obs2/kg resulted in the growth of Portland cement strength by 47 and 18% at 2 and 28 days of hardening, respectively.The increase of fineness of GGBFS from 310 to 500 mThe kinetics of strength development of Portland cements and concretes incorporating GGBFS and FA is lower at the early age of structure formation, but accelerates at later ages due to the pozzolanic reaction that takes place.The synergetic effect of the compressive strength development was revealed in Portland cement and concrete containing granulated blast-furnace slag and fly ash.The replacement level of clinker in cements and Portland cement in concretes with GGBFS and FA depends on their specific surface.The obtained results show the efficiency of unactivated fly ash usage for the cement and concrete production and the necessity of permanent control of fly ash fineness to determine the replacement level of the clinker in a cement and a cement in a concrete.Simultaneous use of supplementary cementitious materials of different nature activity such as GGBFS (latent hydraulic) and FA (pozzolanic) allows for the improvement in the microstructure of concretes due to the formation of an extra amount of ettringite and tobermorite-like low-basic calcium hydrosilicates as well as hydroaluminosilicates in the non-clinker part of cement paste. Unactivated fly ash also allows the particle size distribution to be optimized in cements and concretes to archive the prolong time of cement hydration, which results in obtaining a more refined and compact structure over time due to its role both as a mineral addition with pozzolanic properties and as a microfiller. As a result, the durability and the stability of rebars in such concretes increase, which conforms to the strategy of sustainable development in the construction.Incorporation of cement replacement materials such as GGBFS and FA in concretes results in the reduction of water absorption from 5.5% to 4.7% and, as a result, in the increase of resistance to freezing and thawing damage, improving its durability. The decrease in compressive strength reduction was observed from 30.2 for concrete C to 24.1% for concrete CS10(380)F10(510) after 100 freeze–thaw cycles.Concretes containing 20% by mass of SCMs will have a better corrosion resistance in comparison with the concrete on the basis of Portland cement CEM I. Steel reinforcement protection does not worsen in concretes incorporating 10% by mass of GGBFS and 10% by mass of FA due to the low Portland cement substitution rate and their tighter microstructure. The obtained results concerning the corrosion susceptibility of reinforcing steel bars in concrete incorporating GGBFS and FA will be taken into account in the future developments of this study in the real reinforced concrete elements.The experimental work was done to evaluate the influence of a granulated blast-furnace slag and fly ash on the properties of Portland cements and concretes as well as the corrosion susceptibility of the reinforcing steel bars. The above-mentioned recycling waste was used for a partial replacement of binding materials in this article. The following conclusions can be drawn from the study:


IgG4-related disease is an immune-mediated systemic inflammatory condition characterized by tissue infiltration of IgG4-positive plasma cells and elevated serum IgG4 concentrations. Peripheral neuropathy is an atypical manifestation of this disease. We describe an unusual case of vasculitic neuropathy in a patient with IgG4-related kidney disease. A 55-year-old woman presented with right leg weakness progressing to bilateral leg weakness, pain and numbness of the legs, and impaired gait. She was previously evaluated for weight loss and anemia with a CT scan of the abdomen due to concern for malignancy. Abnormal enhancement of the kidneys was seen, and laboratory work-up and kidney biopsy were consistent with IgG4-related disease. Myeloperoxidase-antineutrophil cytoplasmic antibodies were also positive. In combination with the patient’s asymmetric leg weakness and painful neuropathy, this raised concern for vasculitis. Sural nerve biopsy confirmed vasculitic neuropathy. Recent studies have demonstrated an overlap in the clinical characteristics of IgG4-related disease and the anti-neutrophil cytoplasmic antibody-associated vasculitides, which are known to cause vasculitic neuropathy. Clinicians should recognize this association, and IgG4-related disease should be considered in the differential diagnosis in patients with peripheral neuropathy in the right clinical context.  IgG4-related disease (IgG4-RD) is an immune-mediated systemic inflammatory condition characterized by fibrosis and IgG4-positive plasma cell infiltration of affected tissues and is often associated with elevated serum IgG4 concentrations. First described in patients with autoimmune pancreatitis, it has since been shown to affect multiple organs including kidneys, aorta, lacrimal glands, and salivary glands . NeuroloA 55-year-old woman with a history of allergic rhinitis and asthma presented with right leg weakness, which then progressed to bilateral leg weakness. She reported 14 kg of unintentional weight loss over the past year, and laboratory evaluation revealed iron deficiency anemia. This was concerning for malignancy and prompted CT scan of the abdomen that demonstrated bilateral enlargement of the kidneys with heterogeneous contrast enhancement and loss of normal corticomedullary differentiation. Kidney biopsy showed storiform fibrosis and plasma cell-rich interstitial inflammation with IgG4 immunostaining showing clusters of IgG4-positive plasma cells , suggestThe patient was subsequently seen in neurology clinic prior to initiation of immunosuppressive therapy. By this time, her leg weakness had been present for 8 months, and she also reported 6 months of burning pain and numbness of her lower extremities. Examination was notable for bilateral weakness of ankle dorsiflexion and plantarflexion, loss of pinprick and vibration sense distal to the ankles, and a tentative gait. Electrodiagnostic testing showed a length-dependent sensorimotor axonal polyneuropathy. Given the initially asymmetric pattern of leg weakness, the severity of her pain, and positive MPO-ANCA, there was concern for vasculitic neuropathy. Biopsy of the left sural nerve was performed, which showed severe myelinated and unmyelinated fiber loss in all fascicles, a recanalized epineurial blood vessel, and dense perineurial mononuclear cell infiltrates, consistent with vasculitic neuropathy. IgG4 immunohistochemical staining demonstrated a few scattered IgG4-positive plasma cells . BaselinThe patient was treated with prednisone 40 mg daily and had resolution of neuropathic pain and improvement in gait within 4 weeks. Prednisone was tapered off over a 6-month period. She also received IV immunoglobulin 0.5 g/kg/day for 4 doses then 0.4 g/kg weekly for 8 weeks, followed by rituximab 1 g every 6 months. Subsequent examination demonstrated improvement in lower extremity strength and normal gait. At 6 months of therapy, serum IgG4 level normalized to 57.1 mg/dL, IgE level decreased to 283 IU/mL, and ESR normalized to 25 mm/h. Her kidney function remained normal. The patient’s combination of histopathologic findings, radiographic findings, and elevated serum IgG4 level meet criteria for definite IgG4-RD based on proposed comprehensive diagnostic criteria as well as organ-specific diagnostic criteria for IgG4-related kidney disease , 4. VascANCA-associated vasculitides (AAV), which include microscopic polyangiitis (MPA), eosinophilic granulomatosis with polyangiitis (EGPA), and granulomatosis with polyangiitis (GPA), are systemic vasculitides commonly associated with neuropathy that can have clinical characteristics similar to IgG4-RD, and should be considered in the differential diagnosis of IgG4-RD . Recent In one study, researchers examined sural nerve biopsy specimens of 149 patients with neuropathy who had clusters of inflammatory cells on histopathology. 29 patients had IgG4-positive cell infiltration on histopathology, of which 20 also had elevated serum IgG4 levels. 23 of the 29 patients were previously diagnosed with a primary vasculitic neuropathy such as MPA or EGPA . Another case report described a patient who was diagnosed with EGPA in the setting of multiple mononeuropathies and purpura, with skin biopsy showing leukocytoclastic vasculitis. The patient also had enlarged lacrimal and submandibular glands and was concomitantly diagnosed with Mikulicz’s disease, a manifestation of IgG4-RD . Other sThe relationship between IgG4-RD and vasculitic neuropathy has not yet been fully delineated. The patient in this case met criteria for definite IgG4-RD, and also had a vasculitic neuropathy with positive MPO-ANCA. Clinicians should recognize the possibility of this co-occurrence and understand that positive ANCA does not exclude a diagnosis of IgG4-RD, and likewise, an elevated serum IgG4 level does not exclude a diagnosis of AAV. In addition, IgG4-RD should be considered in the differential diagnosis in patients with peripheral neuropathy and evidence of systemic inflammatory disease. None. All authors have nothing to declare.


We also demonstrated the presence of c-Met+ CTLs in mouse tumors [+ CTL populations arise only under conditions caused by a pathological microenvironment. Hence, this particular c-Met+ population is barely detectable in tumor-free [+ T cells are able to expand only after activation or in pathological settings. Based on these findings, we wondered whether c-Met expression can be induced on CD4+ T lymphocytes from human peripheral blood mononuclear cells (PBMCs) upon T cell receptor (TCR) triggering.The receptor tyrosine kinase c-Met is essential for embryonic development and tissue regeneration, as it promotes cell survival, proliferation, migration, and angiogenesis , 2. The e tumors  and cente tumors . Interesmor-free  and naïvmor-free , suggest+ T cells among freshly isolated PBMCs from healthy donors (HDs). Although Th17 and Th17.1 populations exhibited increased mRNA expression of c-Met, no or very weak c-Met protein levels were detected in the different CD4+ Th subsets as assessed by flow cytometry  and together with IFNγ  than did the CD4+c-Met- subset , LPAM (α4β7), and LFA-1 (αLβ2), three major integrins expressed by T cells, after anti-CD3/CD28 activation of PBMCs from four HDs. Flow cytometry showed increased expression of the Itgα4 subunit (CD49d) by the CD4+c-Met+ subset compared to the corresponding c-Met− counterparts. However, no differences were observed in the expression of the other VLA4 subunit, β1 (CD29), the LPAM subunit (β7), or LFA-1 subunits (ItgαL and Itgβ2)  Fig. . Finallyets Fig. .+c-Met+ T cells results in an increased adhesion capacity. We blocked the Itgα4-mediated leukocyte-endothelial cell interaction with a monoclonal antibody, anti-Itgα4 , which is known to inhibit the trafficking of lymphocytes from the blood into the CNS [+c-Met− and CD4+c-Met+ T cells was performed 72 h after activation, and then the selected cells were seeded on a monolayer of HUVECs (in the presence or absence of TNFα) for 1 h. As shown in Fig. +c-Met+ and CD4+c-Met− T cells were able to adhere to TNFα-activated HUVECs. Notably, after anti-Itgα4 blockade, 70% of CD4+c-Met+ T cells (672 ± 75 to 197 ± 25 cells) but only 35% of CD4+c-Met− T cells (535 ± 59 to 337 ± 109 cells) lost their adhesion capacities . The transmigration capacity of CD4+c-Met+ cells was significantly higher than that of CD4+c-Met− T cells  than for their c-Met− counterparts ; Fig. +c-Met+ T cells have a greater ability to adhere and transmigrate through endothelial cells, at least partially due to the overexpression of Itgα4 on their surface.To investigate the effect of the Itgα4 subunit on CD4lls Fig. . Again, + T cells express c-Met in patients with inflammatory diseases, such as multiple sclerosis (MS) or cancer . We collected PBMCs from HDs, MS patients, and MM patients and stained for CD4+c-Met+ T cells. Immunofluorescence of PBMCs subjected to a Cytospin protocol identified CD4+c-Met+ T cells in MS and MM patients but barely detectable cell counts in HDs  and higher levels of Itgα4 compared to the levels in CD4+c-Met- T cells. Furthermore, anti-Itgα4 treatment directly restrained the adhesion and transmigration capacity of CD4+c-Met+ T cells in vitro. CD4+c-Met+ T cells could be detected directly ex vivo from PBMCs from patients with inflammatory conditions, including MS and MM. Our study offers a perspective of previously unsuspected c-Met signaling in activated CD4+ T cells, which requires further exploration to better understand the functionality and potential clinical applications of targeting these cells in patients with inflammatory conditions.In conclusion, our study uncovered the presence of human c-Met-expressing CD4Methodes and legend Figure S1-3Table S1Figure S1Figure S2Figure S3


Polymer composite reinforcement (PCR) and its use to produce high-quality concrete with the right design and technological and formulation solutions can demonstrate the results obtained with the steel rebars. This article discusses the synergistic effect from the combined reinforcement of concrete with traditional polymer rods and dispersed fiber, which, as a result, lead to an increase in strength and deformation characteristics and an improvement in the performance of compressed and bent structural elements. The synergistic effect of the joint work of polymer rods and dispersed reinforcement is considered in the context of relative indicators , showing the relationship between strength characteristics and concrete density. The behavior of glass fiber in a cement matrix and the nature of its deformation during fracture were studied by scanning electron microscopy. It is shown that the use of PCR and dispersed reinforcement makes it possible to increase the strength characteristics of concrete in bending. In quantitative terms, the achieved results demonstrated that the CSQ values of a beam reinforced with a PCR frame with the addition of glass fiber were 3.4 times higher compared to the CSQ of a beam reinforced with steel reinforcement frames. In addition, for a beam reinforced with a PCR frame with no fiber addition, the CSQ values were three times higher. Consequently, the requirements for materials and structures of buildings being erected have significantly increased ,2,3. Newmissions , the neemissions , to incrmissions , and themissions . The leaIn recent years, considering the development of multi-story construction, the percentage of the use of the so-called polymer composite reinforcement has increased , which wSteel reinforcement does not fully implement its strength potential and is limited by the mechanical characteristics of concrete ,10. It sThus, polymer composite reinforcement is an efficient solution. Many scientists dealing with issues and problems of construction are in search of optimal design solutions, that is, the number, size, and cross-section of rods located in the concrete body ,12,13,14In this regard, an urgent topic for research is the development of new structural, theoretical, and technological solutions for obtaining effective polymer composite reinforced concrete structures for new types of buildings and structures.Significant strengthening of concrete structures ,16,17,183, namely, 3 and 5 kg/m3, reduced the level of damage to the lining structure after seismic impact.Polymer composites have excellent corrosion resistance and strength properties. However, their long-term reliability can be reduced by creep and fatigue . NeverthThe aim of  was to iTo improve flexural strength and impact characteristics, in , thin paBonded reinforcement is commonly used in concrete in civil engineering to increase the load-bearing capacity of a structure or to minimize the negative effects of long-term operation and possible defects. The quality of the adhesive bond between the reinforced structure and steel or composite elements is essential for effective reinforcement. In , a methoReinforced concrete beams made of a fine-grained fiber composite with the addition of steel fiber in the amount of 1.2% of the composite volume were studied in . The resIn , the empThe authors of  investigIn , a compaIn the study , experimAn overview of classical and modern developments from fiberglass in strengthening and restoring civil engineering objects was provided in . The resFrom the review above, it can be seen that only pure reinforcement types were compared, without any mixing or combination in the structural aspect. That is, the reinforced elements were compared only with steel rods or with dispersed fiber. Therefore, in connection with the development and formulation of a working hypothesis, we assume the so-called synergistic effect arising as a result of combined reinforcement with rods and fibers, which, as a result, will lead to an increase in many characteristics and an improvement in the operation of the obtained compressed and bent elements. Furthermore, in the above works, the authors investigated the absolute indicators: the strength of concrete under various types of stress–strain states, deformability of concrete in a stress–strain state. However, the issue of relative indicators, that is, the ratio of strength and density characteristics of the obtained concrete or reinforced concrete elements, has not been studied.Thus, from the literature review and analysis, at least two scientific research deficiencies devoted to concrete enchainment issues with polymer composite reinforcement can be seen. Deficiencies are expressed in constructive and technological aspects.In the constructive aspect, only certain types of reinforcement were previously compared without combining with others. In this sense, the combination of polymer reinforcement bars with fiber seems to be an urgent scientific task.In the technological aspect, the absolute indicators were previously investigated: the strength and deformability of concrete under various types of stress–strain states. However, relative indicators have not been studied, that is, the ratio of strength and density characteristics of the resulting concrete or reinforced concrete elements. Based on this, the study’s goal was to obtain new theoretical knowledge about the work at the microlevel of the cement matrix and fibers and develop ideas about the operation of compressed and stretched elements with an increased coefficient of structural quality made of concrete, combined with polymer composite reinforcement and dispersed fiber. The study’s objectives will be to investigate using SEM analysis the nature of the contact zone of the cement matrix and fibers and directly fibers during destruction to determine the quantitative characteristics of the joint work of a reinforced concrete element at the micro and macro levels, their theoretical justification, and experimental confirmation.The rational composition of concrete, the scheme, percentage of reinforcement with rods, and the rate of fiber reinforcement were adopted following ,29,30,31To manufacture the columns, Portland cement of the PC 500 D0 brand  was used as a binder. Natural crushed stone from quartzite rocks  was used as a large, dense aggregate. The physical and mechanical characteristics of the coarse aggregate are presented in Quartz sand was used as a fine aggregate . The physical and mechanical characteristics of the fine aggregate are presented in For the manufacture of reinforcing cages, polymer composite reinforcement PCR  and steel reinforcement class A400 with a diameter of 6 mm  were used. Comparative characteristics of metal and composite reinforcement are presented in Glass fiber  treated with surfactant was used as dispersed reinforcement; the physical and mechanical characteristics are presented in As a control composition, heavy concrete was designed on dense aggregates of class B30 with the required workability grade P1 (cone draft 1–4 cm). The parameters of the composition of the concrete mixture obtained as a result of calculations are reflected in In total, three series of prototypes were made. Each series included two samples with dimensions of 600 mm × 150 mm × 150 mm. In addition, to control the strength of concrete, cubes were made in parallel with each series of pieces with a size of 10 mm × 10 mm × 10 mm (3 pcs.), and prism samples with a length of 150 mm × 150 mm × 150 mm (3 pcs.) were made to determine the ultimate deformations during axial compression and tension. The first series of pieces included two samples reinforced with Ø6 A400 metal reinforcement. The second series included two samples reinforced with Ø6 polymer composite reinforcement. The third series also included two samples reinforced with Ø6 polymer composite reinforcement with additional dispersed glass fiber reinforcement. Each first sample in the series was tested for short-term central compression, and every second sample in the series was tested for bending.The preparation of the concrete mixture for the prototypes was carried out following the previously established procedure ,33. For -Technological equipment: laboratory concrete mixer BL-10 ; laboratory vibrating platform SMZh-539-220A ;-Testing equipment: hydraulic press PGI-500  with a measurement error of ±1%; tensile testing machine R-50  with a measurement error of ±1%;-Measuring instruments : measuring metal ruler 1000 mm with a measurement error of ±0.5 mm; laboratory scales with a measurement error of ±0.05%; device for measuring deviations from the plane NPL-1; device for measuring deviations from perpendicularity NPR-1.In this study, we used:For the manufacture of prototypes, standard forms of the FP-150 brand  were used. Compaction of the fiber-reinforced concrete mixture during the formation of the samples was carried out on a laboratory vibrating platform SMZH-539-220A with mechanical fastening, the vibration time averaged 60–90 s. The next day after molding, the samples were stripped and placed in a standard hardening chamber for 28 days until the design strength was achieved.A diagram of the reinforcement of the prototypes is shown in The prototypes were tested at the age of 35 days from the date of manufacture. The prototypes were tested on a PGI-500 press. When testing prototype columns for compression, the load was applied step by step in 10–12 stages, with the same increments of longitudinal deformations that made it possible to follow the work of prototypes when the load increased to maximum, and then it decreased on the descending branch until destruction. After each stage of loading, the element was held for 10 min. At each stage, the readings of the devices were taken twice: immediately after increasing the load and after holding before the next stage of loading. Compression tests of specimens are shown in To determine the actual strength characteristics of concrete, simultaneously with the testing of each series of samples, tests of reference sample cubes were carried out. Compression tests of cubes were carried out in accordance with the requirements of GOST 10,180 “Concretes. Methods for strength determination using reference specimens”   MPa/s.In tensile testing, the prism samples were loaded to failure at a constant rate of increase in the load (0.05 ± 0.01) MPa/s.Tests of prisms for axial compression and axial tension were carried out at a constant rate of deformation to obtain the strength and deformation characteristics of concrete and its full deformation diagrams “σ–ε” with descending branches. The measurements of the concrete deformations of the test prisms were carried out by a chain of strain gauges with a side length of 50 mm and dial indicators with a graduation value of 0.001 mm. Axial compression and axial tension tests were carried out following the requirements of GOST 24,452 “Concretes. Methods of prismatic, compressive strength, modulus of elasticity and Poisson’s ratio determination” (interstate standard) .When determining the prismatic strength of concrete, loading the specimen to a load level of 40 ± 5% was performed in steps equal to 10% of the expected breaking load, keeping within each step the loading rate 0.6 ± 0.2 MPa/s. At each stage, the load was held for 4–5 min, and readings were recorded by the instruments at the beginning and at the end of the load stage. At a load level equal to 40 ± 5%, the devices were removed from the prism samples, after which further loading of the sample should be performed continuously at a constant load rate 0.6 ± 0.2 MPa/s.The study of the microstructure was carried out on a VEGA II LMU scanning electron microscope  at an accelerating voltage of 20 kV.Images were obtained using SE and BSE detectors. The SE (secondary electron) detector provides information on the surface morphology of the sample. The BSE detector (reflected or backscattered electrons) provides information on the phase and chemical inhomogeneity of the material (phases and areas with a higher average atomic weight are colored in lighter shades).The surface of the samples was sputtered with metal using an Emitech sputtering device.Based on the results of the experimental studies, the bearing capacity, mass and structural quality factor (CSQ)  of the p-N is the bearing capacity of the column specimens, kN; For sample columns:-M is the bearing capacity of the sample beam, kN × m; For sample beams:The CSQ was calculated using the following formulas:In the analysis performed, the indices of the column specimen reinforced with a steel reinforcement frame were taken as the starting point of reference.The influence of the type of reinforcement used on the mass of the prototype columns and beams. According to the experimental data obtained, the samples reinforced with a frame made of steel reinforcement (up to 5% more mass of concrete samples with PCR) had the highest mass. As for the mass of samples reinforced with a frame made of polymer composite reinforcement without adding fiber and with the addition of glass fiber in the amount of 4% of the cement mass, their masses differed slightly, which is logical and is explained by the content of dispersed fiber.The influence of the type of reinforcement used on the bearing capacity of the prototype columns. According to the experimental data obtained, the highest bearing capacity was possessed by a column reinforced with a frame made of steel reinforcement, and the value of its bearing capacity was 10% higher than that of a column reinforced with polymer composite reinforcement. As for the column reinforced with a frame made of polymer composite reinforcement with the addition of fiber, its bearing capacity increased by 5% in comparison with a column reinforced with a frame made of polymer composite reinforcement without adding fiber.The influence of the type of reinforcement used on the bearing capacity of the prototype beams. According to the experimental data obtained, the highest bearing capacity was possessed by a beam reinforced with a frame made of polymer composite reinforcement with the addition of fiber. The value of its bearing capacity was 3.4 times higher than that of a beam reinforced with a frame made of steel reinforcement. As for the beam reinforced with a frame made of polymer composite reinforcement without the addition of fiber, its bearing capacity was three times higher in comparison with a beam reinforced with a frame made of steel reinforcement.Influence of the type of reinforcement used on the structural quality factor of the prototype columns. According to the obtained experimental data, the column reinforced with a frame made of polymer composite reinforcement without the addition of fiber had the lowest coefficient of structural quality. As for the coefficient of structural quality of a column reinforced with a frame made of steel reinforcement and a column reinforced with a frame of polymer composite reinforcement with the addition of fiber, the values of their structural quality coefficients had insignificant differences.Influence of the type of reinforcement used on the structural quality factor of the prototype beams. According to the experimental data obtained, the highest coefficient of structural quality was possessed by the beam reinforced with a frame made of polymer composite reinforcement with the addition of fiber. The values of its structural quality coefficient were 3.4 times higher in comparison with the coefficient of the structural quality of a beam reinforced with frames made of steel reinforcement. As for a beam reinforced with a polymer composite reinforcement frame without adding fiber, the values of its structural quality factor were three times higher.Analysis of the deformation diagrams obtained from the experimental data revealed the following characteristic features: in fiber-reinforced concrete, in comparison with concrete, the top of the diagrams fits up and to the right, which is explained by an increase in mechanical strength and giving the material a more viscous character of destruction and an increase in the tested ultimate deformations until the destruction of the sample due to the dispersed micro-reinforcement with glass fiber.For polymer composite reinforcement, the relative elongation is directly proportional to the tensile load up to failure, in contrast to steel, which has a zone of elastic work, a yield point, a zone of self-strengthening, and rupture. This means that, when operating under load, the deflections of structures reinforced with polymer composite reinforcement will increase uniformly, up to destruction, in proportion to the increase in external load, in contrast to structures reinforced with steel reinforcement. A feature of the operation of steel reinforcement is the presence of a stage when deflections grow without increasing the external load at the moment of reaching stresses corresponding to the yield point, which is not observed in polymer composite reinforcement ,37,38.The image in This image makes it possible to establish that the fiber undergoes deformation when the sample is broken and pulled out of the body of the cement stone, which is noticeable in the image and is expressed in the change in the thickness of the fiber and characteristic bends visible in the micrograph. Thus, it is obvious that when the sample is deformed, the fiber, in turn, undergoes its own deformation, which, however, is not so significant, and when fractured, it is the fiber that creates the so-called damping effect and imparts a viscous character of fracture to the cement matrix. Therefore, using these fibers makes it possible to smooth out the “explosive” nature of the destruction of a rather brittle material—concrete, especially high-strength concrete.This SEM image is used to support the hypothesis that the fibers tend to break rather than pull out (which is often a problem with inappropriately used fibers). Observing the ongoing fiber deformation, it can be concluded that the applied fiber was well anchored in the body of the cement matrix and, thus, the selected recipe approach is correct.This can explain and substantiate the need for additional fiber reinforcement in a rational range to give high-strength concrete with high-strength reinforcement a more viscous nature of destruction. Thus, at the micro-level, with the help of a microscope, our working hypothesis is confirmed, and the results of experimental studies of the mechanical properties of the obtained sample are supplemented.The photo  shows thThe fiber in this case is a filler and the cement stone is a matrix. The photograph clearly shows the contact zone between them. At the micro-level, the fiber is also exposed to the adhesion of particles of the hydration products of the cement stone, which suggests a good degree of adhesion between the fiber and the cement stone. Thus, the fiber, which has both the function of a damper and imparting a soft tough character of destruction, is also a fairly integrated component that is not rejected by the cement stone, despite its foreign character in the concrete body, and it confirms a high level of adhesion as well as anchoring in the concrete body.In combination with rod reinforcement with polymer composite reinforcement, the nature of the adhesion of which with a cement stone can be seen with the naked eye due to the fact of its large size, this image  confirmsComparing our study with studies previously conducted by other authors, it should be noted that there are differences in the methodological approach.In the classical approach, authors working with compressed and bending reinforced concrete elements phenomenologically set the reinforcement parameters, usually taking into account only traditional types of steel, differing in diameter and number of rods.If we are talking about the study of fiber-reinforced concrete, then, as a rule, in this case, the authors phenomenologically set the classical parameters of dispersed reinforcement: the percentage of reinforcement, the length of the fibers, and the choice of material is determined by the accumulated experience.Our phenomenological approach was based on the hypothesis that high-strength reinforcement, among other things, does not use its resource to the full due to the fact that concrete takes on a large share of the bearing capacity in reinforced concrete elements. Therefore, our goal in the formation of our phenomenological approach, in contrast to other authors, was the idea to increase the bearing capacity while reducing the weight and, at the same time, allowing the structure of the concrete stone to obtain the required useful deformability due to the dispersed reinforcement and hardening of the reinforced concrete element at the micro level.Thus, we are considering our phenomenological model, which is based on two assumptions.It is possible to simultaneously reinforce concrete elements both with rods, that is, at the macro level and with dispersed fiber, that is, at the micro-level.In this case, an important factor is the material for the manufacture of fibers and reinforcing bars.Thus, our research is a pivotal basis for the subsequent development of research in the direction of numerical modeling, the operation of rods from various materials in combination with dispersed reinforcing fibers, also made from various materials.-Comparison by type of reinforcing elements;-Comparison by type of stress–strain state.As part of a comparative analysis of our research with research by other authors, two main criteria can be identified:None of the analyzed works ,29,30,31Thus, the scientific novelty of the study is the application of combined reinforcement of heavy concrete with polymer composite reinforcement in combination with fiber. Such studies have not previously been conducted. It resulted that the technology of combined concrete reinforcement with new lightweight types of reinforcing components provides a synergistic effect, significantly bringing the characteristics of such an element to the characteristics of a traditional reinforced concrete analogue. At the same time, the structure’s weight was reduced considerably, opening up the prospects for the use of such lightweight and durable elements in the construction of new high-rise and large-span buildings and structures.Several factors can explain the resulting synergistic effect.Firstly, the main reason for the increase in the coefficient of constructive quality (up to 240%) of the combined reinforced element is a reduction in its mass (up to 5% with the adopted reinforcement scheme) compared to the reinforced concrete analogue. The mass of the element reinforced only with polymer composite reinforcement was not that much inferior to the mass of the combined reinforced element (up to 0.7%).Secondly, the strength characteristics of the concrete itself, that is, the element that receives compressive loads, increased significantly. Thus, concrete reinforced with dispersed fiber acquired greater structural strength under central compression, and the fragile nature of its destruction was prevented.Thirdly, the additional fiber reinforcement of the bending element allowed the concrete to unload to a greater extent directly on the polymer composite reinforcement, which alone carries this function for simple types of reinforcement, only polymer composite or steel reinforcement.Fourthly, due to the proposed recipe constructive solutions, an increase in the bearing capacity of the combined reinforced element was achieved up to 13% in relation to concrete reinforced only with PCR rods.Additional evidence confirming the hypotheses about the synergistic effect was the SEM analysis, the plotted stress–strain diagrams for the compression and tension of concrete, the physical experiments with the determination of absolute indicators and the calculations of the relative indicators.All this ultimately leads to a sharp increase in the coefficient of the structural quality of the reinforced element.Based on the performed theoretical review and experimental research results, the following conclusions can be drawn.Polymer composite reinforcement and concrete based on it, with the correct design and technological and recipe solutions, can approach the same performance to those obtained when using steel reinforcement and, at the same time, according to the studied indicators of the mass of the structure, its bearing capacity and the coefficient of structural quality. It was revealed that polymer composite reinforcement is still inferior to the used steel reinforcement in terms of bearing capacity in its pure form. Still, it significantly wins in terms of the indicator “weight constructions”. In quantitative terms, this is expressed as follows: the CSQ values of a beam reinforced with a PCR frame with the addition of glass fiber were 3.4 times higher in comparison with the CSQ of a beam reinforced with steel reinforcement frames, and the CSQ values of a beam reinforced with a PCR frame without adding fiber three times higher.In this regard, in modern conditions of dense urban development and engineering-geological conditions in which they are forced to erect high-rise and large-span buildings and structures, this characteristic will be a great advantage. However, also according to the results of a theoretical review and experimental studies, it was revealed that an additional recipe, technological or constructive solutions are needed when reinforcing elements made of concrete. This solution turned out to be the combined reinforcement of heavy concrete with polymer composite reinforcement and dispersed fiber. Thus, in a qualitative aspect, the combined reinforcement had a significant effect, and when applying such a solution, it is possible to approximate the characteristics of polymer composite reinforcement and concrete based on it to those of traditional reinforced concrete.In quantitative terms, this is expressed as follows: reinforcement with polymer composite reinforcement according to the previously presented scheme and the percentage of fiber reinforcement was 4% of the cement mass. Under this condition, the coefficient of the constructive quality indicator was almost identical to the traditional reinforced concrete.The SEM analysis performed while observing the fiber deformation revealed that the applied fiber was well anchored in the body of the cement matrix and, thus, the selected recipe approach is correct.Therefore, the resulting developments can be recommended in the practice of design and technology and construction of buildings and structures.


Development of sustainable concrete as an alternative to conventional concrete helps in reducing carbon dioxide footprint associated with the use of cement and disposal of waste materials in landfill. One way to achieve that is the use of fly ash (FA) as an alternative to ordinary Portland cement (OPC) because FA is a pozzolanic material and has a high amount of alumina and silica content. Because of its excellent mechanical properties, several studies have been conducted to investigate the use of alkali-activated FA-based concrete as an alternative to conventional concrete. FA, as an industrial by-product, occupies land, thereby causing environmental pollution and health problems. FA-based concrete has numerous advantages, such as it has early strength gaining, it uses low natural resources, and it can be configurated into different structural elements. This study initially presents a review of the classifications, sources, chemical composition, curing regimes and clean production of FA. Then, physical, fresh, and mechanical properties of FA-based concretes are studied. This review helps in better understanding of the behavior of FA-based concrete as a sustainable and eco-friendly material used in construction and building industries. It was also reported that annually OPC production releases about 1.5 billion tons of CO2 globally, which is corresponding to about 9% of the worldwide total CO2 emission globally . In. In70]. On the other hand, in a cyclone furnace, up to 70–80% of the total ash that is retained as boiler slag from the crushed coal is utilized as fuel , while m2O3, SiO2 , and CaO; and these core minerals are constituents in coal-bearing rock strata under different guidance standards used for FA quality assurance , CaO (1–2.5%), and C4A3S (1–2.5%) than in low-calcium FAs . The. The5]. 2 ratio of the C-S-H produced in mixes containing FA was less than those in mixes with cement [Furthermore, FA particles exhibit a similar phenomenon to the glass. It was illustrated that glass particles were shielded with fibrous hydrates layer, signifying C-S-H’s propensity to nucleate on glass surfaces. In addition, the C-S-H formed from hydrating FA cement pastes is comparable to the cement substitution by metakaolin and slag, with a high surface area and a foil-like morphology . The CaOh cement .The total heat produced by hydration of OPC and mixed pastes with FA in 2 days is shown in It was also reported that the decrease in the total heat evolution with incorporating FA can be because of the relatively low specific surface area and low solubility of the aluminosilicate present in FA .The construction industry is one of the world’s fastest-growing sectors. According to statistics, approximately 260 billion tons of cement is needed to meet various global construction needs . Over thResearchers have identified key applications for FA in the future based on the yearly time series data by recognizing a pattern of development using regression analysis ,275 and 2 emissions in the cement production. FA-based concrete exhibit high pozzolanic activity, opposed to their mineralogical characteristics and fineness properties. Most of the existing studies did not consider the effect of main influential parameters, particularly particle size distribution, particle packing effect, and alkaline activator solutions, on the strength development of FA-based concrete. FA is potentially utilized to substitute a significant volume of OPC (up to 50%) without influencing concrete durability. However, using 50% FA as a replacement by the cement weight may negatively affect the concrete strength. FA’s fineness has a significant role in concrete because it affects the workability and the rate of pozzolanic activity in concrete, thereby contributing to the enhancement of the properties of concrete. FA-based concrete demonstrated a superior performance to that of OPC-based concrete in resisting sulfate attack, acid attack, and carbonation. To approve the favorable influences of FA on concrete characteristics and durability problems, the following research directions are recommended for future studies.–The manufacturing and improvement of the performance of FA-based AAM must be controlled, and the reaction aspects of the material should be studied in detail. To this end, several facets, for instance, kinetics, thermodynamics, sympathies of intermediates and perceptions into their systems, and the grades to which the Si-O-Al are polymerized and oligomerized, should be studied. These will develop progressively improved performance of the concrete when the extra additives or components are involved. However, further research is needed to confirm that the manufacturing–structure–behaviors correspondence is accurate.–The majority of FA-based AAMs are stiff and susceptible to cracking. This performance obliges restrictions in applications and influences the long-term durability of AAMs. Therefore, innovations in the preparation must be applied to produce improved FA-based AAM composites.–Currently, FA-based AAMs are only formed at the research laboratory scale with empirical formulations. Thus, several studies on FA-based AAM production are required and must endeavor to adopt FA-based AAMs on a large scale.–2 remained unsatisfactory. However, shifting the guidelines for preparation is worthy of further investigation.The performance of FA-based AAMs for immobilization, toxic metal adsorption and the sealing of CO–As an alternative material to conventional concrete, FA-based AAM may be endowed with unique properties or additional functionalities. Therefore, novel applications of FA-based AAMs are worth discovering. For example, FA-based AAMs with biomass can be approved as new light-weight and incombustible materials.–The potential use of FA in producing high-strength and self-consolidating concretes must be studied.–Fibers must be used to increase the strength and longevity of FA in the concrete hardened state.–The use of FA in the design of eco-friendly buildings and cities should be highlighted.The use of FA as an SCM in concrete could resolve the disposal and health issues induced by the generation of ash due to coal combustion in industrial boilers or electric utility. The use of FA in concrete could also help lower the pollution induced by the cement factories by decreasing the CO