Ultra-high-performance liquid chromatography coupled with Q-Orbitrap mass spectrometry (UHPLC-Q-Orbitrap-MS), performing targeted and non-targeted approaches , were used to analyze the samples. The dissipation of flutriafol was fitted to a biphasic kinetic model, with a persistence, expressed as half-life (t1/2), lower than 17 days. During suspect screening, three metabolites  were tentatively identified. Unknown analysis led to the identification of four additional metabolites . The results revealed that the proposed methodology is reliable for the determination of flutriafol and its metabolites in courgette and tomato, and seven metabolites could be detected at low concentration levels. The highest concentration of metabolites was found in the laboratory conditions at 34.5 µg/kg . The toxicity of flutriafol metabolites was also evaluated, and some of them could be more toxic than the parent compound.Flutriafol is a systemic triazole fungicide that is used to control diseases in various crops. A study was developed to evaluate the metabolism and dissipation of flutriafol in two different scenarios: laboratory and greenhouse conditions. Courgette and tomato samples treated with a commercial product (IMPACT Spain has over 50,365 ha of greenhouse used for horticultural production, and most of them are located in the Southeast region. The province of Almeria is known for having the largest land coverage of greenhouses in the world, reaching 48,389 ha in 2018 [To control crops, plant protection products (PPPs) are widely used in agriculture to combat various diseases . They cot1/2), of flutriafol in soil is 1177.3 and 1587 days in field and laboratory conditions, respectively [The persistence, expressed as half-life : triazole alanine (TA), triazole lactic acid (TLA), triazole acetic acid (TAA) and 1,2,4-triazole  . The degThe only investigation that monitored TDMs from flutriafol and the parent compound in field conditions was performed on apple samples, detecting TA and TAA at trace levels (<0.001 mg eq/kg), while 1,2,4-T and TLA were not detected . Even thDue to the lack of a comprehensive understanding of the behavior and dissipation kinetics of flutriafol and its degradation into its known and unknown metabolites, it was necessary to study flutriafol in two different scenarios. For that purpose, trials were carried out under laboratory (over 43 days) and greenhouse (over 55 days) conditions in courgette and tomato samples, respectively. The monitoring period in the greenhouse study was longer than in the laboratory because in the laboratory trials, courgette samples were treated with the PPP after they were collected, while in the greenhouse experiment, the tomato samples were treated with the PPP while still on the plant.Courgette was selected for the laboratory trials for two reasons. First, to the best of our knowledge, the dissipation of flutriafol in this vegetable has not been reported yet, and, second, courgette has a different shelf-life than tomato. Tomato was chosen for the greenhouse scenario because there is a significant demand for this crop in Europe and it is the most important crop in Almeria, Spain . TherefoThis study is the first one to evaluate the metabolism and dissipation of flutriafol in vegetables in two diverse scenarios, providing kinetics parameters of this fungicide and detecting known and unknown metabolites, taking advantage of the use of HRMS. The findings of this study are relevant for understanding the overall behavior of flutriafol in vegetables.Flutriafol  was supplied by Dr. Ehrenstorfer . The triazole acetic acid compound was purchased from Supelco  (CAS registry No. 28711-29-7) with a purity of ≥98%. An individual stock standard solution (1000 mg/L) was prepared by dissolving 10 mg of the pure compound in methanol (10 mL). Intermediate solutions (10 mg/L and 1 mg/L) were prepared in methanol.® EVO (12.5% flutriafol (w/v), SC, suspension concentrate) was purchased from FMC Agricultural Solutions, S.A.U. , and it was manufactured by Cheminova A/S .Methanol and acetonitrile, both LC-MS grade (99.9% of purity), were purchased from Honeywell Riedel-de-Haёn . Formic acid (>98% of purity) was obtained from Panreac AppliChem  and water, LC-MS grade, was acquired from J.T. Baker . A mixture of acetic acid, caffeine, Met-Arg-Phe-Ala-acetate salt and Ultramark 1621 (ProteoMass LTQ/FT-hybrid electrospray ionization (ESI) positive) from Thermo Fisher Scientific  were used for the accurate mass calibration of the Q-Orbitrap analyzer. The commercial pesticide product IMPACTThermo Fisher Scientific Vanquish Flex Quaternary LC  was used for chromatographic analysis.A Hypersil GOLD™ aQ column , supplied by Thermo Fisher Scientific, was employed. The flow rate was set at 0.2 mL/min, the injection volume was set at 10 μL and the column temperature was set at 30 °C. The mobile phase was composed of a water solution of 0.1% formic acid (eluent A) and methanol (eluent B) . The elu2, 95%), 35 (arbitrary units); auxiliary gas , 10 (arbitrary units); S-lens RF level, 50 (arbitrary units); capillary temperature, 300 °C; and heater temperature, 305 °C. The mass spectra were acquired using four alternating acquisition functions: (1) full MS, ESI+, without fragmentation  collision cell was switched off), mass resolving power = 70,000 full width at half maximum (FWHM), AGC target = 1 × 106; (2) full MS, ESI−, without fragmentation  collision cell was switched off), mass resolving power = 70,000 FWHM, AGC target = 1 × 106; (3) data independent mass spectrometry fragmentation (DIA-MS/MS), ESI+ , mass resolving power = 35,000 FWHM, AGC target = 2 × 105; (4) DIA-MS/MS, ESI− , mass resolving power = 35,000 FWHM; AGC target = 2 × 105. The mass range in the full-scan experiments was set to m/z 60–900 [The LC system was coupled with a hybrid mass spectrometer, Q-Exactive Orbitrap Thermo Fisher Scientific , using the electrospray interface (ESI)  in positive and negative modes. Different ESI parameters were set: spray voltage, 4 kV; sheath gas  to confirm the fragment ions by in silico approach. Furthermore, MassChemSite version 3.1  was used to identify unknown metabolites.Fresh courgettes were purchased from a local market in Almeria (Spain). The study began immediately upon arrival at the laboratory. Dissipation experiments were performed from November to December 2021 under normal sunlight (at least 8 h sun exposure) in laboratory conditions.® EVO commercial product was 0.09%.To simulate normal crop conditions, courgettes were treated with the PPP at the recommended dose. Additionally, a higher application dose was evaluated to study the effect of the dose on the dissipation of the compound as well as to detect metabolites that may be present at low concentrations. Therefore, two concentration levels were tested: single dose (SD) and double dose (DD), which was two times the recommended dose. The single rate for the IMPACTSamples were stored at room temperature (15 ± 5 °C) and were analyzed in triplicate with an approximate size of 500 g at 2 and 6 h, and at 1, 2, 5, 10, 15, 22, 29, 36 and 43 days. Some courgettes were established as blank, and they were not treated with the commercial product.Furthermore, water loss was controlled by checking the weight of two courgettes , and they were considered in the concentration calculation of the dissipation process.2 in area) was divided into four blocks or plant lines (39 plants for each block). Three of them were sprayed with a 12.5% flutriafol emulsion at the recommended dose, and the other one, which was used as control, was not irrigated with the commercial product. Each block was divided into three plots, resulting in three replicates. The greenhouse study was carried out in an agricultural greenhouse, using a hydroponic system located in the province of Almeria (Spain) over a period of 55 days. The field had not been treated with flutriafol or other fungicides with similar structures to triazole fungicides in the past five years. During the sampling period, from January to March of 2022, only this triazole fungicide was applied. The working field . They were then analyzed using the solid-liquid extraction (SLE) procedure previously published . This meC(t)) and the half-life time (t1/2) were calculated using the biphasic kinetic model (Equations (1) and (2)). In these equations, C0 and C1 represent the concentration (µg/kg) in sample (courgette or tomato) at time t (day), C0 represents initial concentration of flutriafol (µg/kg) and k1 and k2 are the dissipation kinetic rate constants (days−1).The dissipation of flutriafol was evaluated by plotting residue concentration of this fungicide against time. The residual concentration , version 5.1.2,  was empl® EVO commercial product) were evaluated. Flutriafol was monitored and its concentrations were measured at each sampling time and some differences were observed. It was noted that the general behavior of this fungicide during the study was an increase in concentration until it reached a maximum level and then decreased, as shown in The natural behavior and dissipation profile of flutriafol . Different kinetic models  were tested, and the biphasic model provided the best fit  was almost half the value of DD (C0 = 689.1 µg/kg). However, as shown in k1 and k2 values) were reached: 0.05 and 0.06 days−1, respectively, for the SD, and 0.04 days and 0.07 days−1, respectively, for DD, with similar correlation coefficient values ), indicating similar behavior in both scenarios.The lowest concentration of flutriafol in laboratory trials was observed six hours after treatment at both application doses. In this context, a concentration value of 407.9 µg/kg was observed in the case of the SD and 685.9 µg/kg when the DD was tested. In terms of initial concentration values, these revealed different information. As expected, the initial concentration of the SD (t1/2) was similar for the two doses. For the SD, the t1/2 was 14.8 days (k1) and 14.0 days (k2), whereas for the DD, the t1/2 was 16.3 days (k1) and 14.9 days (k2)  , indicatBased on these results, in which the dissipation process exhibited a similar trend, few differences were observed between both concentration levels under laboratory studies. Therefore, the applied dose of PPP did not affect the dissipation of flutriafol.2 = 0.966), the following values were obtained: the initial concentration (C0) of flutriafol was 315.6 µg/kg, the t1/2 was 8.9 days, and the degradation rates were 0.08 and 0.09 days−1, for k1 and k2, respectively, demonstrating few differences at the beginning and the end of the dissipation process (In relation to the greenhouse study (R process . Flutria2 ≥ 0.9708) was fitted to field tomato and cucumber samples treated with 25% flutriafol SC at DD [2 > 0.9385) in grape samples treated with 25% flutriafol SC [−1 in tomato and cucumber samples [−1 in grape samples [−1, which is a similar value to that obtained in the first study, while a slightly higher value was obtained in the second one, although the matrix was also different. Therefore, the degradation of flutriafol in grape was faster than in tomato samples. In terms of half-lives, 9.2–10.2 days [It is worth noting that the dissipation process of flutriafol in vegetables did not conform to the few previous publications on greenhouse studies. In one of these studies, first-order kinetics , the laboratory samples were sprayed with a double dose (DD) to thoroughly monitor the metabolic pathway of flutriafol and evaluate any differences between the two concentration levels using a ‘suspect analysis mode’. The main reported metabolites of flutriafol were TDMs: TA, TLA, TAA and 1,2,4-T, and their chemical structures are shown in m/z 70.03997, which corresponds to the compound 1,2,4-T, was observed as the common fragment ion of two of these metabolites; TA and TAA, with −1.42 and 0.25 ppm of mass error, respectively, indicating adequate values. This ion is characteristic of triazole fungicides, and it is the common part of the structure of all of them [For each metabolite, at least one fragment ion was monitored using Qual Browser and compared with the information provided by Mass Frontier™ software. The fragments were sorted according to the following criteria: the most abundant ion; retention time, which must be equal to the corresponding precursor ion; and mass error (lower than 5 ppm). Suitable mass errors for the fragment ions, with values between −1.96 and 2.28 ppm were achieved . It shou of them .The results obtained in the laboratory trials showed that the three TDMs  were identified at different times during the study: TAA was the first to be detected, as it was found the first day after the application of the commercial product; TA was detected 2 days after treatment; and, finally, TLA was detected on the 15th day. There were significant differences between the results of the SD and DD experiments, as most of the compounds could only be detected when the DD of commercial product was applied, indicating a low concentration of the majority of the metabolites in the studied samples.In relation to the greenhouse experiment, TA was the first compound found, and it was detected 2 days after application, followed by TAA (day 7) and TLA, which was only identified 24 days after application.Since a commercial analytical standard of TAA was available, the compound was successfully confirmed  by compaOnce the known metabolites were identified in the samples, their concentration was estimated. Therefore, a semi-quantitative approach was performed using the parent compound (flutriafol) for all metabolites, except for TAA, whose analytical standard b was avaIn the laboratory trials, the concentration of the TDMs was found to be between 2.9 µg/kg for TAA (24 h after application) and 34.6 µg/kg for TA (at 48 h). TA, which showed the highest value of all TDMs, was detected at the beginning of the study . TAA was found one day after the treatment with the PPP at very low levels (2.0 µg/kg) and its concentration decreased over time. Regarding TLA, this metabolite showed concentration levels lower than the limit of quantification of flutriafol (LOQf), which was set at 2 µg/kg, both for the SD and the DD (on days 15 and 43 of the study). Therefore, it was noted that the behavior of TDMs decreased as the study progressed, and most of the compounds could only be identified (<LOQf) but not quantified.The concentration of TDMs in greenhouse trials was 3.6 µg/kg for TAA on day 7. The other two metabolites (TA and TLA) could only be identified on one day of the study  and were not quantified due to the levels being below LOQf.In conclusion, TDMs were found at trace levels in both the laboratory and in greenhouse scenarios, except for TAA, which was detected and quantified at concentrations between 2.0 and 3.6 µg/kg in both experiments. TA was also identified at levels > 30 µg/kg in laboratory conditions.According to a study carried out by EFSA on TDMs in apple samples, in which 1,2,4-T and TLA were not identified and in which TA and TAA were detected at trace levels (<0.001 mg eq/kg) , consideIn terms of TDMs toxicity, the most toxic compounds detected in this study, were TA and TLA, whose ARfD value = 0.3 mg/kg bw, although they are less toxic than the parent compound: ARfD = 0.05 mg/kg bw .m/z 302.10994) and its corresponding peak and retention time (7.42 min), the tool proposed different compounds with their own retention times, structures and exact masses. The following criteria was used to select unknown metabolites: appropriate chemical structures in comparison with flutriafol. As a result, a total of four compounds were identified as new metabolites of flutriafol: C16H11F2N3 , C16H14F2N4 , C19H17F2N5O2  and C22H23F2N3O6 , as shown in 2). MF3 may have been formed through acetylation of a urea binding, and MF4 through glycosylation of the benzene ring, as shown in An unknown screening was performed to identify unknown flutriafol metabolites that have not been reported by EFSA or in the literature. For that purpose, the raw files obtained for each experiment were processed using MassChemSite . This soThe most relevant characteristic parameters of these metabolites are shown in 19H17F2N5O2 (m/z 386.14231) and C22H23F2N3O6 (m/z 464.16170), respectively, were identified in both experiments.It should be highlighted that MF1, MF3 and MF4 were detected under laboratory conditions, and that MF2, MF3 and MF4 were detected in the greenhouse experiment. This may be due to the different matrix used in each case or the differences in agronomic factors. Therefore, MF3 and MF4, which correspond to CAfter these metabolites were tentatively identified, a semi-quantitative estimation was carried out, using the parent compound as the standard, to determine the concentration of these compounds.In both laboratory trials (SD and DD), the unknown metabolites were identified at the end of the study, revealing that MF3 and MF4 were only detected at levels below LOQf. Regarding MF1, this compound increased its concentration from 2.7 (day 29) to 2.9 µg/kg (day 43), as the study progressed for the SD , and froThe detected metabolites in the greenhouse scenario are shown in 50) in oral rats. For MF1 and MF3, these values were 313.8 and 944.2 mg/kg, respectively. No values were set for MF2 and MF4. Based on these values and the LD50 of flutriafol being set at >1140 mg/kg [Finally, in order to evaluate the possible impact of flutriafol metabolism, the toxicity of the metabolites was evaluated. As the information for flutriafol and TDMs has been reported in literature, as described above, the toxicity of the unknown metabolites was estimated by predicting their lethal dose 50%  of flutriafol was less than 17 days, and it was observed that the dissipation process was not significantly affected by the application doses used in this study.For the first time, the dissipation of flutriafol was investigated in two vegetable samples under two different scenarios: laboratory (courgette samples) and greenhouse (tomato samples) conditions. The persistence (19H17F2N5O2 and C22H23F2N3O6) being detected in both scenarios, highlighting the role of HRMS in these types of studies. Therefore, this study has successfully provided valuable information about the metabolism and dissipation of flutriafol in vegetables, describing an overall picture of this fungicide in two matrices under two different conditions, increasing the knowledge about triazole fungicides and related metabolites.The results confirmed that the proposed analytical methodology is effective and reliable for the detection of flutriafol and seven metabolites: three known compounds (TDMs) and four unknown compounds. All TDMs could be suitably identified in the samples except for 1,2,4-T. These metabolites were present at trace levels, with concentrations below LOQf in most cases. The unknown metabolites were described for the first time, with two of them (C


Due to technological advancements, concrete can be currentlyproducedwith varying strengths and durability based on its intended use. However,in many applications, concrete still needs to be improved in termsof its mechanical and physical properties. The addition of fibersto concrete is one of the most widely used methods for improving itsmechanical and physical properties. The study focuses on the effectsof the high aspect ratios and reinforcing indexes on the mechanicalproperties of the hybrid and non-hybrid chopped glass fiber reinforcedconcrete (CGFRC). In this study, the glass chopped fibers (GCFs)  with four different volume fractions and four different lengths  were mixed into the concrete considering the aspect ratios between200 and 2800 and the reinforcing indexes between 1 and 42. A totalof 51 samples were prepared for the study that included 3 control,36 non-hybrid, and 12 hybrid samples. Then, the flexural strengthand compressive strength tests were conducted on the CGFRC samples.To obtain detailed information about fiber pullout, fiber breakage,debonding, or cracking in the matrix, digital microscopy and scanningelectron microscopy examinations were performed. The flexural strengthof the hybrid samples increased with the higher aspect ratios andreinforcing index values, whereas the flexural strength of non-hybridsamples decreased with the higher aspect ratios and reinforcing indexvalues in the CGFRCs. Moreover, all non-hybrid and hybrid CGFRC sampleshad lower compressive strengths than the control samples in termsof compressive strength. With an increase in the fiber volume fraction,the mixing and workability of the samples considerably decrease, andthe increase of the fiber volume fraction caused brittle fracturesin concretes to be transformed into ductile fractures. Because steel rebars areheavy construction materials, their mass increases the forces on thestructures. Hence, researchers have recently been experimenting anddeveloping new strengthening materials for concrete structures byusing different engineering materials such as chopped steel fibers,fiber-reinforced polymers, steel plates, and ferrocement.5 Today, adding additives to concrete or reinforcing concrete withdifferent materials is one of the most preferred methods to improvethe mechanical properties of concrete due to its significant advantagessuch as easy application and minimum change in the overall size. Especially,improving the mechanical and physical properties of concrete by addingchopped fiber to concrete has been a method that has been appliedfor many years. While the use of fiber in concrete was made with goathair, horse mane, straw, and even human hair in the past, the useof chopped fibers such as steel, carbon, glass, aramid, or basalthas currently become widespread.10 The improvement of concrete performance by using fibers dependson many factors such as fiber type and size, aspect ratio, reinforcingindex, and fiber–matrix interaction.11 Besides, the compatibility of fibers with components of the concreteis very significant for the structural performance of the concrete.Despite its high compressivestrength, concrete is a brittle materialwith low tensile strength. Therefore, cracks and possible failuresof concrete structures occur when tensile stress exceeds the correspondinglow strength.12 Therefore, most previousstudies have concentrated on the benefits of chopped fibers on themechanical performance of concrete. Yuan and Jia13 conducted some experiments to study the effect of polypropylenefibers and glass fibers on the mechanical and microstructural propertiesof the concretes. To prepare the concrete samples, different ratiosof water/binder (0.30 and 0.35) and different fiber contents  were used. The study concluded that thewater/binder ratio affected the glass fiber ratio. Furthermore, theglass fibers were more effective at absorbing water in concrete thanpolypropylene fibers. Cakir14 investigatedthe effects of two types of chopped fibers which were CGFs and choppedbasalt fibers on the physical and mechanical properties of polymermortars (PMs). In the study, three different mixtures were preparedand mechanical tests were conducted on the samples based on differentcuring times: 7, 14, 21, and 28 days. According to the experimentalstudies and evaluations, chopped fibers influence the mechanical propertiesand failure modes of PMs, and CBFs are better additives than the CGFs.Moghadam and Izadifard15 studied the effectsof steel and glass fiber addition on the behavior of normal concreteat high temperatures. In the study, the fiber volume fraction was0.25% and experimental temperatures ranged from 28 to 800 °C.At high temperatures, steel fibers improved compressive, tensile,and shear strengths in a range of 9–27, 8–198, and 1–22%,respectively. In specimens containing the glass fiber, the compressionand tensile strength were both improved by 1–18 and 19–213%,respectively. Ganta et al.16 conducteda study to determine how fiber type and aggregate content affect thehardened and durability properties of self-consolidating concrete.In the present experimental program, steel, glass, and steel/glasshybrid fibers were tested. Based on the results, the hybrid reinforcedself-consolidating concrete shows good mechanical and durability performancecompared to other mixtures. Ali and Qureshi17 examined the effects of glass fibers on concrete with recycled coarseaggregates (RCAs) concerning mechanical performance and durability.The results of testing show that 50% RCA concrete outperforms theplain natural coarse aggregate (NCA) concrete at 0.5% GF in overallmechanical performance .Test results indicated the 50% RCA concrete outperforms the NCA at0.5% GF in terms of overall mechanical strengths which were compressive,split tensile, and flexural strength. Riad et al.18 assessed the behavior of reinforced concrete beams underdifferent fire and cooling conditions by adding discrete glass fibers.In this study, a series of 18 concrete beams with different compressivestrengths were tested to analyze the behavior of RC beams containingdiscrete glass fibers under various cooling and fire conditions. Finally,the study recommended that the percentage of discrete glass fibersin concrete should not exceed 0.5%. Kizilkanat et al.19 studied the effect of basalt and glass fibers on high-strengthconcrete. Several experiments were conducted using concrete samplesproduced with different fiber volumes .The fibers reduced the workability of the concrete but did not haveany significant effect on its modulus of elasticity. The compressivestrength of the samples also increased slightly with fiber additions.In addition, the study pointed out that fiber additions enhanced flexuralstrength. Tassew and Lubel12 investigatedthe effects of the CGFs on the mechanical and rheological propertiesof the ceramic concrete binder with phosphate cement. It was shownthat by increasing fiber content in the mortar mixture, hardness,bending, and shear strength increase, but compressive strength andelasticity are not affected. In addition, the fiber reinforcementdecreased the workability of the mortar mixture.Chopped glass fiber (CGF) is one of the most preferred choppedfibers today. In comparison to other chopped fibers, the CGF has gainedsubstantial attention from the engineering community as a result ofits low cost, high tensile strength, high chemical resistance, andinsulating properties for large and small concrete structures alike.Vf = 0, 0.5, 0.75, and 1%) and four differentlengths  weremixed into the concrete considering the aspect ratios (Lf/ϕ) between 200 and 2800 and the reinforcing indexesbetween 1 and 42.Based on theprevious literature, it is clear that many studieshave been conducted on the use of fibers in concrete to enhance itsphysical and mechanical properties. However, the studies conductedare still insufficient and there are still unresolved or inconclusiveissues concerning the improvement of fiber materials for concrete.Therefore, the study focuses on the effect of the high aspect ratiosand reinforcing indexes on the flexural and compressive strengthsof the CGF reinforced concrete (CGFRC). For this purpose, 51 CGFRCsamples, having 3 control, 36 non-hybrid, and 12 hybrid samples, wereconstructed, and then, three-point bending strength and uniaxial compressivestrength tests were conducted on the samples. In this study, the GCFs with four different volume fractions. The resultsof the experiments are presented in Because silicasand has a high hardness value, resistance to abrasion and weatherconditions, and chemical stability,2.1.2For the preparation of thesamples, Portland cement was used as a binder material. The cementused in this study was CEM I/42.5R cement from Akçansa CementCompany. The chemical and physical characteristics of the cement arepresented in 2.1.3Lf) was preferred as 3, 6, 12, and 24 mm. Technical properties of theCGFs are given in The glassfiber produced by Dost Kimya was used as the chopped fiber to focuson the effect of the CGFs on the mechanical properties of concretesamples. In the study, the fiber length (2.2Lf/ϕ), fiber volume fractions (Vf), and reinforcing indexes (RIs). The RI is consideredas a key factor when determining how fiber content and aspect ratioaffect composite properties.25 The CECS3826 defines a RI as the characteristic value of fiber, as followsVf, Lf, and ϕ represent the volume fractionof fiber, fiber length, and fiber diameter, respectively. The useof 23 and Cao et al.24 reported a new formula for reinforcing indexwhich is as follows.Several factorsinfluence the structural behavior of the CGFRCs, including the geometricaland mechanical properties of the CGFs, aspect ratios  ratio was preferredas 0.50% in all samples. The GCFs  with four different volume fractions  and four different lengths  were mixed into concrete withaspect ratios (Lf/ϕ) of 200, 400,800, and 1600. According to previous studies, the use of 0.25% glassfiber did not have a significant effect on the bending and compressivestrengths19 and the bending and compressivestrengths decreased when the glass fiber content exceeded 1.00%.27 In this study, a range of values that offerbetter results has been selected to assess the mechanical behaviorof the fiber producing the additive. To achieve uniform distributionand prevent damage from over-mixing, the components were mixed bya mechanical stirrer for 90 s, with 60 s at low speed and 30 s athigh speed .gh speed 1.For the flexural and compression tests, the freshGCFRC mixtureswere placed into the 40 × 40 × 160 mm prism molds. Eachmold was then vibrated on a shaking table in order to compact thesamples. The prepared samples were placed in a curing cabinet for24 h in molds. After removal from the molds, the samples were placedin a curing cabinet at 20 °C and 90% relative humidity for 28days. A total of 17 sets (51 pieces) of 40 × 40 × 160 mmsized rectangular prism samples were prepared for the experimentalstudies. 325 The experimental studiesstarted with determining the densities of each sample, followed bythree-point bending tests. According to the Test Procedure (Item 9)section of TS EN 196-1,25 each part ofthe sample divided into two parts was subjected to uniaxial compressivestrength tests to determine the compressive strength of each part.ach part4. All ex4Scanning electronmicroscopy (SEM) and digital microscopy (DM)examinations of the samples were conducted to better understand thefracture process, failure mode, bonding mechanism, and debonding orcrack morphology. 1600× DM (Bushman 1600×) and Sigma 300SEM (Zeiss) were used for visual inspections. Visual inspections wereconducted on the specimens that failed the mechanical testing by cuttingthem through the cross-section of the failed surface. During SEM inspection,samples were coated with gold to prevent electrons from charging them.To obtain the desired magnification, a 5 kV accelerating voltage wasapplied.53. It was determined that WPUV values showeda general decreasing trend with the addition of fiber but did notshow a significant change  withfour different volume fractions  and four differentlengths  were mixed into the concrete consideringthe aspect ratios between 200 and 2800, and the reinforcing indexesbetween 1 and 42. A total of 51 samples were prepared for the studythat included 3 control, 36 non-hybrid, and 12 hybrid samples. Then,the flexural strength and compressive strength tests were conductedon the CGFRC samples.Experimental studies have demonstratedthat fiber reinforcementhas both positive and negative effects on the mechanical propertiesof concrete. Specifically, CGF reinforcement has a positive effecton the flexural strength of concrete, but a negative effect on itscompressive strength. In hybrid samples, the flexural strength increasedwith higher aspect ratios and reinforcing index values, whereas non-hybridsamples did not experience this increase. In terms of compressivestrength, all CGFRC samples have lower compressive strengths comparedto the control sample. Additionally, it is determined that the mixingand workability of the samples considerably decrease with an increasein the fiber volume fraction, and the increase of the fiber volumefraction causes brittle fractures in concretes to be transformed intoductile fractures. In most samples, interface cracks, fiber breakage,fiber and aggregate pull-outs, fiber agglomerations, and air voidsare common problems. These problems, however, become more intenseand negative as fiber ratios increase.Although it is expectedthat the study will contribute significantlyto the literature, it is essential to conduct similar studies on sampleswith low fiber ratios. Moreover, the study considers only CGF andcertain fiber sizes and fiber ratios. Due to these reasons, it isrecommended to conduct additional studies with different types offibers, different fiber lengths, and different fiber ratios.


Geopolymer (GP) concrete is a novel construction material that can be used in place of traditional Portland cement (PC) concrete to reduce greenhouse gas emissions and effectively manage industrial waste. Fly ash (FA) has long been utilized as a key constituent in GPs, and GP technology provides an environmentally benign alternative to FA utilization. As a result, a thorough examination of GP concrete manufactured using FA as a precursor (FA-GP concrete) and employed as a replacement for conventional concrete has become crucial. According to the findings of current investigations, FA-GP concrete has equal or superior mechanical and physical characteristics compared to PC concrete. This article reviews the clean production, mix design, compressive strength (CS), and microstructure (Ms) analyses of the FA-GP concrete to collect and publish the most recent information and data on FA-GP concrete. In addition, this paper shall attempt to develop a comprehensive database based on the previous research study that expounds on the impact of substantial aspects such as physio-chemical characteristics of precursors, mixes, curing, additives, and chemical activation on the CS of FA-GP concrete. The purpose of this work is to give viewers a greater knowledge of the consequences and uses of using FA as a precursor to making effective GP concrete. CO2 emissions estimated between 85 to 105 gigatons could be emitted into the atmosphere over the next 33 years because of the production of PC [2 emissions ensuing from PC production would hinder the world from achieving its sustainability goals and result in more detrimental impacts on the environment. Thus, this study aims to explore alternative sustainable materials such as GP concrete that can be used in place of PC concrete [The most utilized construction material is concrete because of its performance, cost, ability to form different shapes, and availability of raw materials locally ,2,3. Howproduced ,7,8,9. Ion of PC ,10,11. Ion of PC ,11. Figuconcrete .2 emissions [2 emission by about 22–72% [GP concrete is deemed the next generation of concrete because of its sustainability benefits as it offers an innovative way to totally eliminate PC as a binder in the manufacture of concrete, thereby resulting in a substantial reduction in COmissions ,14,15. Gmissions . The alumissions . Figure missions ,19,20, tmissions . Studiest 22–72% ,24,25,26t 22–72% , high-tet 22–72% , and acit 22–72% ,30,31,322O⋅(n)SiO2-type silicates or both. M in the MOH indicates the alkali-metal such as K, Na, and Ca in the activator, which is generally sodium hydroxide (NaOH), sodium carbonate (NaCO3), potassium hydroxide (KOH), or sodium sulfate (Na2SO4) [GPs are composed of the alumino-silicate precursor which is composed of amorphous silica and alumina alongside an activator that acts as the dissolving agent. Geopolymerization is the process by which zeolitic-like materials are converted into a three-dimensional (3D) alumino-silicate gel ,36,37,38(Na2SO4) . In addi(Na2SO4) . Figure (Na2SO4) ,42,43,44This study aims to analyze and answer a lot of worries concerning the viability of FA-GP concrete as the next-generation sustainable concrete. Despite these numerous advantages, the impact of FA on the long-term characteristics of GP concrete remains a source of worry that must be fully understood before FA-GP concrete can be considered a sustainable alternative. In a summary, the purpose of this work is to give viewers a greater knowledge of the consequences and uses of using FA as a precursor to making effective GP concrete ,46,47.3 of FA is utilized for every ton of FA-GP concrete [Depending on their local availability, use, and production cost, various types of precursors have been used to produce GPs ,36,48. Tconcrete . Becauseconcrete ,50. As tconcrete . This reThis study aims to analyze and answer a lot of worries concerning the viability of FA-GP concrete as the next-generation sustainable concrete. Numerous investigators that presented evaluations on FA-GP concretes discovered that the strength characteristics of the FA-GP concrete might be equal, if not considerably higher, compared to those of PC concrete ,55. FurtGeopolymerization is a chemical reaction between alumino-silicate precursors and an alkaline component that results in a hardened product at room temperature or occasionally at an increased temperature . Alkali-In Equation (1), M is a symbol of the alkali-metal cation, n is a symbol of the gradation of polycondensation, w is a symbol of the quantity of chemically attached water molecules, and z is a symbol of the silicon to an aluminum ratio. The GP structures are classified into three varieties based on the value of z: -Si-O-Al-O- , -Si-O-Al-O-Si-O- , and -Si-O-Al-O-Si-O-Si-O-  ,62,63. FIn comparison, five distinct chemical processes exist for the GP, as illustrated in 2, Al2O3, and CaO [FA is an abbreviation for “pulverized fuel ash” generated after the collection of boiler flue gas from coal power plants, as shown in  and CaO , and the and CaO ,75,76,77 and CaO . SEM-EDS and CaO  as shown and CaO ,81,82,834+ and Al3+ molecules occurs because of FA–alkali reactions, followed by more intricate nucleation, oligomerization, and polymerization, culminating in a novel alumino-silicate-based polymer with a 3D network structure. In experiments or applications, the produced FA-GP concrete is placed in a mold and dried in an oven at a specific temperature or at room temperature for a specified period to form the structure indicated in The formation of FA-GP concrete is based on the breakdown of alumino-silicate in FA, which is accelerated by alkali, followed by polycondensation. Manufacturing is supposed to be more energy- and resource-efficient since reactions can occur at low temperatures. The actual responses that occur during the process, on the other hand, are exceedingly complicated and remain a mystery. Condensation between the resulting Si3+ and Si4+ species, which combine to form nuclei and alumino-silicate oligomers composed of SiO4 and AlO4 tetrahedra. Based on the S/A ratio, the networks in alumino-silicate oligomers can be polyciliate -Al-O-Si-chain, polyciliate siloxo -Al-O-Si-Si-chain, or polyciliate disiloxo -Al-O-Si-Si-Si-. Al3+ partly replaces Si4+ in alumino-silicate monomers, and the resulting negative charge in the alumino-silicate bonds is neutralized by alkali cations such as K+ and Na+ [Alkali activation of FA is expected to be critical for geopolymerization formation: in an alkaline sol, the silica, alumina, or alumino-silicates in FA hydrolyze, and the -Si-O-Si- or -Si-O-Al- bonds of alumino-silicate break, releasing active Al and Na+ ,86, as i and Na+ . It has  and Na+ ,89. TablVarious parameters such as particle size, S/A ratio, amorphous composition, and phase composition affect the structure and characteristics of GPs . Precurs2/Al2O3, temperature curing, supplementary water, the molarity of Na2O/Al2O3, and NaOH ratio have a critical influence on the CS of FA-GP concrete [2SO3/NaOH ratio has a substantial impact on GP concrete strength growth. Other studies [GP concrete’s CS is heavily influenced by its content and mix design. As demonstrated in concrete ,115,116.concrete  observedconcrete ,120,121.concrete  and Songconcrete  demonstr studies ,132,133 The creation of a standard for GP concrete is currently in its early stages. June , Ferdous2O3 percentage is around 10 to 20% by weight, while the CaO percentage is less than 5% [The low-calcium FA was used in all experimental trials indicated in  than 5% ,112,138. than 5% .2SiO3 sol. Aluminum oxides and silicon contribute to over 80% of the total weight, with a silicon-to-aluminum ratio of almost 2 for low-calcium FA. The binder is the primary distinction between GP concrete and PC concrete. The GP paste is made by reacting Al2O3 and SiO2 within FA with an alkaline activator sol, which bonds unreacted components to the GP concrete [As in PC concrete, fine and coarse aggregates can be used as aggregates in GP concrete ,127. Theconcrete . Phoo-ngconcrete  proposed2SiO3 sol to NaOH sol by mass results in higher CS of GP concrete and that increasing the workability of GP concrete by incorporating superplasticizer, up to 4% FA by mass, improves the workability of GP concrete; however, this may hurt the CS of GP concrete. It was also discovered that as GP concrete’s water content increases, its workability increases.The amounts and characteristics of the basic ingredients used in the creation of GP concrete influence its workability and CS. According to Pimraksa, Chindaprasirt, Rungchet, Sagoe-Crentsil, and Sato , increasFA-GP concrete CS is affected by S/A ratios, alkali sols, calcium content, additives, and curing conditions.4+ and Al3+ from FA during geopolymerization. A dosage of alkaline sol is typically advantageous for achieving high CS [The dosage and type of alkaline sol affect the generation of Si high CS ,164,165; high CS . For ins high CS  looked i2SiO3 sol is generally combined with NaOH [2SiO3 with a high viscosity can aid in the production of GP gels, resulting in a compact final FA-GP concrete Ms. Furthermore, the activation technique affects the CS of FA-GP concrete. For instance, Rattanasak and Chindaprasirt [4+ and Al3+ species, followed by Na2SiO3 for one minute to assist in generating a homogeneous GP paste. The strength of FA-GP concrete increased as a result of the independent activation. To boost CS, Naith NaOH . This isaprasirt  first in3+ and Si4+. Furthermore, the efficient S/A ratios have a substantial impact on the dissolving, hydrolysis, and condensation reactions of GPs. Condensation action in a high S/A system leads to the predominance of the silicate species itself, generating oligomeric silicates, which condense with Al(OH4)4 and create GP structures of poly (Si–O–Al–O–Si–O–Si–O–) and poly (Si–O–Al–O–Si–O) as shown in 3+ and Si4+ [The FA-GP concrete structure is mostly determined by the S/A ratio generated by the dissolving of Aland Si4+ ,173,174.2. The use of nano-alumina and nano-silica as additives in FA-GP has been found to improve its mechanical characteristics. Phoo-ngernkham, Chindaprasirt, Sata, Hanjitsuwan, and Hatanaka [2+, Na+, and K+) are present, reacting phases including Al and Si have disorganized structures and can generate acidic -Si-OH (silanol) groups with water. The dissolution behavior of vitreous aluminum silicate-containing network modifiers is depicted in The function of additives is to create a crucial equilibrium in oxide dosages to the required level using various add-ons ,177,178.Hatanaka  created Hatanaka  and YangHatanaka  found th2SiO3/NaOH (SS/SH) ratio dosage in the FA precursor.Calcium has been shown to interact with silica and alumina gelation during the geopolymerization reaction, altering the Ms of FA-GP concrete and hence the CS ,193. TheIt is well established that typical GPs require thermal treatment to produce CSs equivalent to or greater than those of PC concrete ,197,198.The type and fineness of the GP raw material are instrumental in developing the strength, durability, and Ms of the resulting GP matrices. The Ms characteristics of FA-GP concrete rely on the FA characteristics, type, and dosage of activators and the curing conditions ,96,98. FThe influence of several SS to SH ratios on Ms studies has been investigated . As showKarthik, Sudalaimani, Vijayakumar, and Saravanakumar  included4.56Si1.44O9.72, quartz SiO2, magnetite Fe3O4, and calcium silicate Ca3SiO5 were recognized as crystalline phases. The inclusion of calcium silicate in the quantitative measurement of fly ash indicates slight cement contamination in Melbourne ash. The binder’s XRD phases are entirely connected to the fly ash source material, and no novel crystalline phases are generated in this binder. Furthermore, the amount of the calcium silicate phase does not decrease with time, indicating that the C3S phase in fly ash does not participate much in the reaction, and the sodium aluminosilicate hydrate (N-A-S-H) gel is still the predominant binding matrix in these specimens. As previously discussed, crystalline phases were revealed after adding an alkaline sol.In reality, Lloyd, Provis, and van Deventer  chroniclAs shown in The study by Aligizaki  showed t+3 for Si+4 in bridging locations, the C-A-S-H matrix chains in the GP system were shown to be longer than the C-S-H gel chains in the PC system [As demonstrated in C system . BecauseC system . The gelTo accomplish the industrialization and commercialization of geopolymers, hurdles must be overcome in three areas: technology, economy, and administration. This also necessitates collaboration between the government, businesses, and the general people for co-governance, which is both an objective necessity for enhancing the quality of the ecological environment and a practical prerequisite for constructing an ecological civilization. Geopolymers derived from solid wastes are expected to become more frequently employed when nonrenewable resources are depleted. Basic research should be prioritized in order to minimize the cost and increase the workability of geopolymers. Based on fundamental research, embarking on solid wastes to explore the preparation of varied characteristics of geopolymers in order to produce high-value-added application domains should be one of the primary paths of future geopolymer research development ,269,270.2 emissions. The use of GP concrete would also result in an effective way to manage industrial wastes such as FA, thereby promoting the sustainable development of society. Discussions presented in this review paper showed that FA-GP concrete could be an alternative to PCC. Nevertheless, this will only happen if both an efficient raw material supply chain and a product supply chain network are in situ. From this perspective, the recent market is optimistic, but it will take time to establish GP concrete as a commercial product worldwide. Desirable performance for diverse engineering applications may be achieved with correct selection and adjustment of GP concrete composition. Regardless of the differences in the precursors’ characteristics, the production and formation and the characteristics of the FA-GP concrete products are highly influenced by the physical and chemical characteristics of FA, AAs, additives, and curing conditions. Based on the discussion in this paper, the following conclusions can be made:The recommended S/A ratio for producing GPs of appropriate strength varies depending on the nature and content of the raw material. S/A ratios of 2–2.5 (class F FA) and 2.5–3.5 (class C FA) were determined to be optimal. The influence of Al and Si on setting and hardening characteristics in high-Ca FA is substantial.2SiO3, are very good and economically viable alkali activating sols. Microstructural investigations revealed that GP concrete prepared with a lower NaOH to Na2SiO3 ratio reacts more efficiently because of the large surface area available for reaction and binding without being clogged by excess water. However, the hydroxyl groups tend to condense when specimens are heated to high temperatures.The conventional AAs, a combination of NaOH and NaFA-ITZ GP concrete consists of gel, pores, fissures, and unreacted FA particles. N-A-S-H gel can improve the strength of the ITZ by promoting the combination of the aggregate interface and the GP matrix. As a result, FA-GP concrete has nearly no obvious weak ITZ near the aggregate.The existence of C-S-H gel, as well as GP gel, improves the mechanical and Ms characteristics of precursors either having high Ca or mixed with Ca components.2 and CaO content should be between 15% and 20%.When it comes to GP synthesis, the criticality of precursor oxide dosage cannot be overstated. In general, the Si oxide content should be between 45% and 55%, the Al dosage should be between 22% and 28%, and the FeCOThe experiment using MIP analysis reveals that extending the curing duration of GP specimens is useful in minimizing pores. The increased geopolymerization limits the connection between the pores, leading to a denser pattern. Furthermore, FA particles and gel may be used to refine the pores in FA-GP concrete. As a result, FA-GP concrete gel pores and capillary pores are concentrated in the tiny pore-size region. However, it is worth noting that the silicon content of the AA plays an essential role in the refining of the FA-GP concrete pore structure.It was found that increasing the fineness leads to an increase in the reaction rate and therefore requires a minus time of heating to achieve strength, as it was found that more than 60 MPa can achieve the strength of GP concrete by increasing the fineness of the precursor material.FA-GP concrete with a compact and denser structure shows high mechanical strength and good efflorescence that make it an ideal choice for the construction industry and has been effectively used in precast industries. Moreover, there is an enormous possibility of utilizing high-Ca GP as a restorative material.With the main alumino-silicate precursors used in the production of GP concrete being industrial wastes, such as FA, the utilization of GP concrete in place of the conventional PCC can be used to reduce COThough FA-GP concrete is a relatively new, innovative, and sustainable engineered material combination with numerous advantages, some difficulties remain, and some are recommended below for further investigation:Currently, the findings of relevant investigations on the Ms of FA-GP concrete are discordant in several respects. For example, the conclusions on forming a new crystalline phase prior to and during the AA reaction of FA are inconsistent. As a result, additional systematic and in-depth research is required to prove this substance’s usefulness with greater certainty. Additionally, advanced analytical techniques such as micro- or nano-analysis and nuclear magnetic resonance must be employed.Suitable guidelines for the selection of aggregate contents in FA-GP concrete should be developed. It is also critical to develop design procedures for each substantial predecessor. Furthermore, the usage of dune sands, fibers, marble dust, date ash, and other materials as precursors should be investigated.Detailed investigations, such as those relating to the derivation and modeling of reaction kinetics under different treatment and production circumstances of the developing class of GP raw material, such as blended GPs and biomass ash GPs, are needed.GP binders need a high pH and heat curing. As a result, efforts are needed to create a room-temperature-cured one-component GP system that uses solid activators rather than alkaline sols in order to gain widespread acceptability in the field.FA-GP concrete should be given more functionality instead of using it as an alternative cement only. FA-GP concrete with biomass has the potential for development as a new class of lightweight fireproof composites.Lastly, more research is needed to evaluate the cost of GP concrete compared to that of standard concrete. Furthermore, contradictory findings regarding the embodied energy and carbon footprint of GPs in comparison to traditional Portland cement must be addressed.


Immunoglobulin G4-related disease (IgG4-RD) is a newly defined disease entity, with great heterogeneity among IgG4-RD subgroups with different organ involvement patterns. Identification of the proteomic characteristics of IgG4-RD subgroups will be critical for the understanding of the pathogenic mechanisms of IgG4-RD.In this study, we performed proteomic analysis using Tandem Mass Tags (TMT) technology with “high field” mass analyzer with improved resolution and sequencing speed to investigate the proteomic profile of saliva and plasma samples from ten untreated IgG4-RD patients and five healthy controls (HCs). Differentially expressed proteins (DEPs) were identified by “t test” function in R package. Functional enrichment analysis was used to investigate pathways enriched in IgG4-RD samples.Most salivary DEPs identified in IgG4-RD patients compared with HCs were mainly enriched in neutrophil mediated GO bioprocess. Within the comparisons between four IgG4-RD subgroups, more DEPs were identified in the comparison of Mikulicz group and Head and neck group. Among four subgroups of IgG4-RD, Head and neck group showed the most distinctive proteomic expression pattern when compared with HCs. Moreover, “Neutrophil mediated process” related GO bioprocess was commonly identified between comparisons of Mikulicz group and Head and neck group, Head and neck group and Retroperitoneal aorta group, Head and neck group and HCs, IgG4-RD patients with saliva gland involvement and those without saliva gland involvement. Key DEPs that involved in this GO bioprocess were identified. Besides, we performed proteomic analysis for plasma samples between ten IgG4-RD and five HCs and there were several DEPs identified overlapped in saliva and plasma.We identified multiple processes/factors and several signaling pathways in saliva that may be involved in the IgG4-RD pathogenesis. All IgG4-RD patients who fulfilled the 2011 comprehensive IgG4-RD diagnostic criteria  were recSaliva was collected between 8–10 a.m. All enrolled IgG4-RD patients and HCs were refrained from eating, drinking, and oral hygiene procedures for at least 1 h before saliva collection. Before collection, a 1 min oral rinse with distilled water was performed and ~5 mL of saliva was then collected in the next 5 min as described . The colIn this study, the Tandem Mass Tags (TMT) technology was applied to investigate the proteasome of saliva and plasma samples . For saliva samples, fraction supernatants were loaded on an UHPLC system (Thermo Scientific™ UltiMate™ 3000) equipped with a trap and an analytical column, and peptides separated from nanoHPLC were subjected into the tandem mass spectrometry Orbitrap Exploris 480  for DDA detection . Scans wFor plasma samples, the ProteoMiner Protein Enrichment Kit  was applied to deplete the high abundance proteins. Then, trypsin or Glu-C digestion was performed on two parallel batches of the samples. The resulting peptides were separated and detected using a Q-Exactive tandem mass spectrometer equipped with an Orbitrap analyzer (Thermo Fisher Scientific) . In brievia the iProX partner repository .Protein identification and quantification were realized by software MaxQuant . The maspository  with thehttps://david.ncifcrf.gov/). Pathways and GO terms with p ≤ 0.05 based on Fisher’s Exact Test were considered as statistically significant. Protein expression patterns were indicated by heatmaps, which were performed using R packages such as “gglot2” and “Complex Heatmap”. The immunologically relevant list of genes curated with functions (https://www.immport.org/shared/genelists). The STRING database (https://string-db.org/) was used to analyze the protein-protein interaction (PPI) network.To identify the differentially expressed proteins (DEPs) between the IgG4-RD subgroups with different organ involvement pattern, we used ‘t test’ function in R package (version 3.6). The thresholds for differential analysis of DEGs were FDR ≤ 0.05 and |Ratio| >1.2. Functional enrichment annotations for GO and KEGG pathways were analyzed using the Database for Annotation, Visualization and Integrated Discovery online tool  DEPs in saliva . By usinWe performed differential protein analysis among different IgG4-RD subgroups: (A) Mikulicz group, (B) Hepatobiliary group, (C) Head and neck group and (D) Retroperitoneal aorta group, and we obtained a list of DEPs from each comparison. In general, there were much more DEPs identified in the comparisons of Mikulicz group or Retroperitoneal aorta group versus Head and neck group than in other comparisons Table 3.Knowledge of IgG4-RD in head and neck regions is important, since the salivary gland is one of the most frequently involved sites . In thisAiming to explore proteomic pattern of IgG4-RD saliva, we performed differential protein analysis between IgG4-RD subgroup and HC samples and obtained a list of DEPs from each comparison in saliva Table S3Saliva gland was reported as one of the most commonly involved sites of IgG4-RD , so we nIn this study, we also collected plasma samples of ten IgG4-RD and five HC for analysis.There are more DEPs detected in plasma than those in saliva (295 up-regulated and 188 down-regulated) Table S4Common upregulated and downregulated DEPs with same expression tendencies identified in the saliva and plasma of four subgroups of IgG4-RD patients were listed in At present, the high heterogeneity of IgG4-RD presents a considerable challenge in etiological understanding, in part because of their heterogeneous manifestations. Despite progresses in the diagnosis and treatment of IgG4-RD, the exact pathogenic mechanism of IgG4-RD with four distinctive phenotypes remains unclear . SeveralIn this paper, most saliva DEPs in IgG4-RD compared with HC belong to the antimicrobials category Table S1We found that “Neutrophil mediated process” related GO BP term that includes neutrophil degranulation (GO:0043312), neutrophil activation involved in immune response (GO:0002283), neutrophil activation (GO:0042119), and neutrophil mediated immunity (GO:0002446), was identified in the comparisons of Mikulicz group with Head and neck group (A vs. C), and Head and neck group with Retroperitoneal aorta group (C vs. D) Table S2We found three upregulated DEPs  and one downregulated DEP (VCP) with expression tendencies overlapped in the saliva and plasma Table 6.Considering SS is another common autoimmune disease that affects salivary and lacrimal glands, thus we performed differential gene expression analysis and functional enrichment analysis using SS and HC data from GSE40568 dataset . As showIn this paper, we compared the DEPs identified in our previously published proteomic plasma data  with theTo our knowledge, this is the first proteomic analysis of saliva samples from four subgroups of IgG4-RD patients. There are also limitations in our study. Firstly, the sample size of our analysis was relatively small; secondly, other types of omics  and validation with laboratory experiments were not carried out in this study and will be investigated in future studies.via the iProX partner repository with the dataset identifier PXD036552 . Datasets of GSE66465 and GSE40568 were downloaded from Gene Expression Omnibus Dataset (GEO Dataset: http://www.ncbi.nlm.nih.gov/geo/).In this study, our proteomics data of saliva and plasma samples from ten IgG4-RD patients and five healthy controls (HCs) were deposited to the ProteomeXchange Consortium The studies involving human participants were reviewed and approved by Tongji Hospital, Tongji Medical College, Huazhong University of Science, and Technology Institutional Review Board. The patients/participants provided their written informed consent to participate in this study.S-YL analyzed the data and wrote the manuscript. TZ made the sample collection. SC and Z-WH provided specific knowledge on IgG4-RD. JZ and LD designed the study and revised the manuscript. All authors contributed to the article and approved the submitted version.This study was supported by grants from the National Natural Science Foundation of China (NSFC Nos. 81771754), and Tongji Hospital Clinical Research Flagship Program (2019CR206).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Concrete is the most widely used construction material. However, it cannot sustain the harsh environment and can easily deteriorate. It results in repair and reworks that amount to a considerable loss of money and time. The life span of concrete reduces if exposed to external attacks, for instance, sulfate attacks, alkali-silica reactions, corrosion, and drying shrinkage. These ubiquitous attacks cause a reduction in service life and raise the need for early repair and maintenance, resulting in higher life cycle costs and structural failures. To resolve these issues, the potential of styrene-butadiene-rubber (SBR) ultrafine powder as cement replacement polymeric admixture at 0%, 3%, 5%, 7%, and 10% have been evaluated. The effect of SBR-powder on concrete is investigated by conducting an alkali-silica reactivity test (ASR), rapid-chloride-permeability test (RCPT), drying shrinkage, and sulfate resistivity tests. Workability, compressive and flexural strength tests are also conducted. For ASR and drying shrinkage, mortar bar samples were cast, exposed to respective environments, and the percentage change in length was measured. For mechanical tests and RCPT, prisms, cylinders and cubes were cast and tested at 28 days. The SBR-powder modification reduces concrete’s permeability, drying shrinkage, and expansions due to ASR and sulfate attacks. SBR powder increased workability by 90%, compressive strength by 23%, and flexural strength by 9.4% in concrete when used at 10% cement replacement by weight. The SBR-powder (10%) modification reduced the RCPT value by up to one-third (67%), drying shrinkage by 53%, ASR by 57%, and sulfate reaction by 73%. Consequently, SBR powder usage can adequately improve the workability, mechanical properties, and durability of the concrete and lead to advanced sustainable concrete with low repair requirements. Concrete is the most widely used construction material, generally composed of cement, crush, and sand. Its benefits and usability usually overshadow its negative impact on the environment. Environmental scientists are continuously struggling to abate the negative impact of cement and concrete production. This research explores expensive carbon capture technologies/sequestration and injecting liquid carbon dioxide to achieve a low carbon footprint.2 emissions and reduce energy consumption, leading to high sustainability and energy efficiency.Sustainable and durable concrete for long-term performance and improved mechanical properties is a dire need of the present and future ,2. ProduThe durability of concrete allows it to resist weathering action, chemical attacks, and physical deterioration without compromising its engineering properties. The durability and longer life span of concrete depend mainly on permeability, i.e., the interconnectivity of concrete pores ,4,5. ItsDurability and service life design have become key factors, especially in extreme environments ,9. ConcrAdmixtures (such as polymers) that can reduce permeability should be explored in extreme environments to resolve durability issues  and incrUltrafine SBR powder is used in the study because it can prove beneficial in reducing permeability. The properties of SBR depend on the styrene and butadiene ratio . It has The presented research aims to evaluate the performance of concrete against environmental issues such as alkali-silica reactivity, drying shrinkage, and sulfate resistivity to ensure long life. Ultrafine SBR powder at 0%, 3%, 5%, 7%, and 10% cement replacement is used. The slump value and compressive and flexural strengths are also determined. SBR powder was expected to improve the durability and physical properties of concrete and mortar. This study implies the solution to the early deterioration of concrete and environmental issues. The production of concrete with low repair and reconstruction demand leads to reduced cement usage, lower carbon footprint, better resources conservation, and higher energy efficiency.The study devises a modified concrete composition, which will not be significantly affected in harsh/ aggressive environments. Due to its intrinsic properties, the selected polymer compensates for the deficiencies in concrete. Thus, the modified concrete can withstand environmental conditions well without repeated repair and reworks and loss of time and money. Otherwise, repairing or replacing existing components due to exposure to an aggressive environment is very expensive and technically difficult .2/g blain fineness was used as a hydraulic binder to produce mortar and concrete samples. Ordinary Portland cement Type-1 conforming to ASTM C-150 with a 3078 cmThe properties of ultrafine styrene-butadiene-rubber (SBR) powder are shown in The mix proportions of concrete for compressive strength, flexural strength, and rapid chloride permeability test samples are given in The mix proportions were selected for all durability tests, as described by corresponding relevant ASTM standards.The concrete samples prepared were 4″ × 4″ × 20″ (100 mm × 100 mm × 10 mm) prisms for flexural strength testing, 2.75″ × 2.75″ × 2.75″ (70 mm × 70 mm × 70 mm) cubes for compressive strength tests, and 4″ × 2″ (100 mm × 50 mm) cylinders for rapid chloride permeability test. The mortar bars for sulfate resistivity, alkali-silica reactivity, and drying shrinkage were 1″ × 1″ × 11¼″ (25 mm × 25 mm × 285 mm) in size. Mortar cubes of size 2″ × 2″ × 2″ (50 mm × 50 mm × 50 mm) were also prepared for compressive strength tests related to sulfate attacks and for water absorption. The compressive strength and flexural strength tests were conducted in accordance with ASTMC109 and ASTMC78. Mechanical strength tests and RCPT tests were conducted after curing samples for 28 days ,27. HoweA slump test was performed to assess the workability of concrete in conformance to the ASTMC143-78 standard. A conical steel mold was used to measure the slump value of concrete . The conFlexural strength test:Higher flexural strength implies better cracks resistance, and it is directly related to the initiation of cracks. Flexural strength tests were carried out on concrete prisms conforming to ASTM C78 . Four prCompressive strength test:Compressive strength tests on four concrete samples for each mix composition were performed with the help of a universal testing machine. Four samples were cast and tested for compressive strength after curing for 28 days. The compressive strength test was conducted in accordance with ASTMC109 .Rapid chloride permeability test (RCPT) and Water absorption test:Permeability is the interconnectivity of concrete pores, which allows the water and other reagents to ingress concrete. The ingression of water and external reagents/ions in concrete causes various reactions and expansions that generate tensile stresses. As concrete has a low tensile strength, it cracks. Cracks, in turn, allow further ingression of ions and lead to higher deterioration . Thus, dPermeability is a crucial factor for durability in a corrosive environment . RCPT teAlkali-silica reactivity test:This test method provides a means of detecting the potential of alkali-silica reactions that causes potentially deleterious internal expansion. The favorable condition for alkali-silica reactions was provided by crushing with reactive silica while preparing the bar samples. The bars were dipped in alkali solution for 14 days at an elevated temperature of 80 °C. The high temperature provided accelerated alkali-silica reaction. The test determined how this alkali-silica reactivity was affected by SBR powder shown in . The tesDrying shrinkage test:The drying shrinkage is affected by temperature, relative humidity, and rate of evaporation. This test method determines the shrinkage/reduction in length of mortar bars that are demolded after 24 h, kept for the next 48 h in a lime water bath, dried, and air stored. The test was conducted in hot, dry weather at 33 °C (91.4 °F) and 57% Relative Humidity (room atmosphere in June). The change in the lengths of mortar bars was measured at 7, 14, 21, and 28 days of their production see . The tesSulfate resistivity test:This test method determines the expansion in length of mortar bars due to sulfate attacks. For sulfate resistivity test, mortar sample of mix ratio as given in SEM was used to study the microstructure of modified concrete on the 28th day of casting. The microstructure of the sample was analyzed by SEM-JEOL-JSM-4680LV at a working voltage of 15 KV. Sample was mounted on a stub of aluminum with carbon tape and was carbon-coated. The sample was carefully examined.The experimental results are shown in the section below.The slump value of concrete increased with the increase in styrene-butadiene-rubber (SBR) percentage, as shown in Flexural Strength:Increasing the SBR powder percentage increased the values of flexural strengths, as shown in Compressive strength:In concrete samples with 0%, 3%, 5%, 7%, and 10% SBR modification, the compressive strength increased gradually with a relative increase in the percentage of SBR powder. The maximum value of percentage increase in compressive strength was 23%, as shown in A previous detailed study conducted by the author revealed that the compressive strength of mortar with SBR liquid latex reduces with an increase in SBR percentage after 5% cement replacement. The reason was the thick layer of polymer stopping further hydration and easy slip-ability between layers of polymers. This study is about SBR powder instead of SBR latex; the compressive strength does not decrease and keeps on increasing with the increasing percentage of SBR powder. The compressive strength kept increasing but is optimum for 5 to 10% utilization.Sulfate attacks and alkali-silica reactions both cause expansion and thus cracking in concrete ,42. In cRapid chloride permeability test (RCPT):Alkali-silica reactivity test (ASR):The most favorable environment for ASR reactions was provided using NaOH alkali solution and aggregates containing reactive silica naturally. Hence, even a slight reduction in mortar bar expansion caused by ASR means that admixture provides better resistance against alkali-silica-reactivity. When alkali from the surrounding, e.g., cement, comes in contact with an aggregate with reactive silica, it forms an alkali-silica gel. So, expansion takes place. Due to the lower tensile strength of concrete, this expansion causes cracking. The mortar bars with 5% and higher cement replacement percentages with SBR powder showed the expansion within the limiting range, i.e., less than 0.2% expansion at 28 days (as suggested by ASTM standard). Overall, 5% of SBR reduced ASR expansion by 32% at 28 days.Alkali silica gel expansions decreased significantly with the addition of SBR powder. Samples with a higher percentage of SBR powder expanded in a limiting range of 0.10% at 14 days and 0.2% at 28 days after casting. The alkali-silica reactivity (expansion) of mortar bars was gradually reduced to an acceptable limit after adding SBR powder; SBR powder positively affects concrete’s durability. The reason for controlled alkali-silica reactions might be the improved microstructure of SBR-powder-modified concrete and decreased permeability, which means SBR modification does not allow much interaction of reactive silica of aggregate with free alkali in cement. SBR powder on hydration blocks nearby alkali from coming in contact with the reactive aggregate. Additionally, SBR may block moisture ingress to the interfacial transition zone (ITZ); water presence is necessary to form a gel and its expansion in ITZ. The ASR expansion of the bar is reduced by 57% at 10% SBR modification. Usually, very expensive lithium salts are used to control ASR. However, ultrafine SBR provides a more sustainable and cost-effective solution against the expansion and cracking caused by ASR.Drying shrinkage:Drying shrinkage is the contraction of the hardened concrete or mortar mixture due to capillary water loss caused by evaporation. It is a major issue in hot and dry weather. The test was conducted at almost 33 °C (91.4 °F) and 57% relative humidity, so shrinkages were relatively higher than at 20 °C (68 °F). SBR reduces the permeability and further loss of water by evaporation. The excellent water retention quality of SBR due to the polymer nature and the blocking of capillaries by producing SBR agglomerates  reduces the evaporation process. Thus, a lesser amount of evaporation helps reduce the shrinkage of bars. Drying shrinkage is reduced by almost 33% and 53% at 5% and 10% cement replacement by SBR powder, respectively.Additionally, the higher tensile capacity  may also cause lesser drying shrinkage cracking and a lower number and size of cracks. In other words, SBR-modified bars might have better resistance against tensile stresses along with a better water-retention property, whereas in control samples, these tensile stresses increase due to shrinkage, leading to cracking, internal wrapping, and external deflection. The results obtained are similar to Moodi’s (2018) findings of SBR’s potential against shrinkage  and dimeSulfate resistivity:Sulfate ingression results in the cracking of the concrete due to ettringite formation in hardened concrete. External sulfate ions in the presence of monosulfates and tricalcium aluminate produce hard needles of ettringite. The hardened concrete is unable to accommodate this expansion of ettringite needles and is cracked. SBR powder reduces the permeability by filling the pores (and reduces sulfate ion ingression in concrete), resulting in remarkably reduced sulfate attacks. Hence, the ettringite formation in hardened concrete due to external sulfate ingression is reduced. Sulfate action-induced expansion is reduced by 33% and 73% using 5% and 10% cement replacement by SBR powder. A similar study found that SBR latex provides excellent resistance against chloride ingress .Water absorption:The water absorption test shown in  shows a Interrelation between Permeability and Durability Issues:Microstructure and pore characteristics due to additives  in concrete greatly influence the mechanical properties, permeability, and durability . Due to The permeability is directly or indirectly related to various properties of concrete, especially durability issues. Following relations are found:The following relations are obtained for the SBR-modified concrete.3A is a major cause of sulfate reactions. Localized SBR powder agglomerates also blocked the capillaries, resulting in reduced permeability. The binding nature (adhesiveness) of SBR powder increased the strength. The non-formation of slippery membranes (unlike latex modification) prevented a reduction in strength that could be caused due to loss of friction. Meanwhile, smaller size hydrates that might be modified and strengthened by SBR powder are a possible cause of higher performance.The workability of concrete increases with the increase in the percentage of SBR powder. SBR (10%) increased slump value by 90%.The compressive strength of SBR-powder-modified concrete increases with increases in SBR percentage. SBR (10%) increased compressive strength by 23%. Moreover, flexural strengths of SBR-modified concrete increase slightly (9.4% for SBR10%) with an increase in the percentage of SBR.SBR powder reduces chloride permeability and provides protection against corrosion caused by chloride ingress. SBR (10%) decreased the charge passed and hence chloride permeability by one-third as compared to the control sample.SBR provides excellent volume stability by giving outstanding performance against dying shrinkage and expansions due to alkali-silica reactions and sulfate attacks. SBR (10%) reduced expansion in mortar bars due to sulfate attacks and ASR by 73% and 57%, respectively, and reduced shrinkage due to ASR by 52%.By increasing SBR powder from 5% to 10%, the durability issues are abated noticeably. The interrelation is developed for permeability and durability issues for SBR-modified concrete.Hence, it is inferred that concrete durability issues are substantially reduced by replacing cement with ultrafine SBR powder from 5 to 10% . A highly sustainable and better-performance concrete is produced that is particularly suitable for harsh environments.This study evaluates the effect of ultrafine styrene-butadiene-rubber (SBR) powder (up to 10%) with a particle size smaller than 85 microns as a partial cement replacement material on concrete issues and attacks in harsh environments. The results corroborate that the fresh and hardened properties of cementitious materials are enhanced significantly by incorporating SBR powder.


Pseudomonas aeruginosa (P. aeruginosa) is a widespread, gram-negative, pathogenic bacterium that causes serious internal and external infections in humans and other animals. The increasing antibiotic resistance has complicated bacterial infection treatment, and current antibiotic therapies cannot cure all infections. Owing to this, bacteriophages (phages) have regained attention as potential therapeutics for bacterial infections. In this study, the phage “PaVOA” was isolated from hospital sewage and characterized. Next, a New Zealand rabbit skin infection model was used to determine the therapeutic efficacy of PaVOA as compared to a phage cocktail or the cephalosporin antibiotic ceftriaxone. Characterization results demonstrated that phage PaVOA belongs to the Myoviridae family, has a double-stranded DNA genome, is resistant to low temperatures (−20°C), is most optimal at 40°C, has good acid–base tolerance, and remains stable for 30 min under 20 W ultraviolet (UV) intensity. The optimal multiplicity of infection of PaVOA was 0.1, and a one-step growth curve showed a short latency period (10 min), thus demonstrating its ability to rapidly kill bacteria. Furthermore, the addition of calcium (Ca) and magnesium (Mg) ions significantly increased the PaVOA titer. An in vivo phage kinetic curve showed that PaVOA was rapidly inactivated within the blood of New Zealand rabbits (undetectable after 12 h), and no animals died due to phage treatment. Wound healing studies showed that the phage cocktail induced a high healing rate and an acceleration of the skin remodeling process, and was more efficacious than ceftriaxone. Therefore, phage cocktail therapy represents a novel therapeutic approach in the treatment of traumatic skin infections caused by multi-drug resistant P. aeruginosa. Pseudomonas aeruginosa (P. aeruginosa), a gram-negative bacterium, is a common opportunistic pathogen in hospitals that can cause serious infections such as pneumonia, urinary tract infections, burn infections, and bacteremia  are viruses that are small in size, do not have a complete cellular structure, and contain only a single nucleic acid. Phages have an estimated population of over 10on Earth  and can on Earth . Importaon Earth . The apphe study , thus suPseudomonas aeruginosa is one of the most prevalent causative pathogens of wound infections in hospitals  and cultured on a shaking table at 37°C for 12 h. Bacterial DNA was isolated using the TIANamp Bacteria DNA Kit . All strains were identified as P. aeruginosa by PCR experiments.Twenty-eight clinically isolated P. aeruginosa drug susceptibility testing provided by the National Committee for Clinical Laboratory Standards (NCCLS) for drug selection. A total of 22 antibiotics , including ciprofloxacin , amikacin , ceftriaxone , meropenem , gentamicin , doxycycline , imipenem , amoxicillin , cefepime , ofloxacin , compound sulfamethoxazole , enrofloxacin , ampicillin , azithromycin , chloramphenicol , tetracycline , nincomycin , norfloxacin , ceftazidime , mezlocillin , cefoperazone , and tobramycin  were used for the antibacterial sensitivity testing of 29 P. aeruginosa strains using the paper diffusion method. The diameter of inhibition circles was used to evaluate the sensitivity of P. aeruginosa to all antibiotics tested.We referred to the recommendations for P. aeruginosa. The double-layer agar plate method was then used to separate phages. Individual phage plaques were chosen for purification once their size and morphology were consistent (purified single phages). One of the lytic phages with transparent plaques was selected, and P. aeruginosa (PA/18) was used as the host bacteria to further study the biological characteristics of the selected phages. Moreover, the host profile of the phage was assessed using the spotting method.Two different bacteriophages were isolated from the sewage of Nanfang Hospital of the Southern Medical University of China. For water sampling, a 0.22 μM filter membrane  was used to remove residual bacteria, followed by centrifugation of the filtered filtrate at 12,000 rpm for 15 min at 25°C, with the resulting supernatant enriched with 11 PFU/mL were loaded onto a copper grid for 1 min, followed by negative staining with uranyl acetate (2% w/v) for 1 min and subsequent aspiration of excess liquid. After drying, the Talos™ F200S TEM  apparatus was used to observe the morphology of phages and to capture images.Purified phage particles with a titer of 10Purified phage concentrates were prepared using the PEG precipitation extraction method . Phage gPseudomonas aeruginosa was cultured in LB broth for 16–18 h. Overnight cultures were then diluted with LB broth to a concentration of 108 CFU/ml. Phages were added at MOIs of 1 × 10−4–1 × 102 and incubated for 4 h at 37°C in a warm oven after shaking and mixing. Cocktails were centrifuged at 12,000 rpm for 10 min followed by isolation of the supernatant to determine phage titers using the double-layer agar plate method. The experiment was repeated thrice.Pseudomonas aeruginosa was cultured to the logarithmic growth phase, at which point the PaVOA phage was added according to the optimal MOI ratio determined previously. Cultures were placed in a 37°C incubator for 15 min prior to centrifugation at 8,000 rpm for 5 min. After discarding the supernatant, sediment was resuspended with preheated sterile LB broth. The suspension cocktail was cultured on a shaking table at 37°C, 200 rpm. Samples were taken at intervals of 10 min in the first 30 min and then every 15 min. Phage titers of samples were determined using the double-layer agar plate method. The experiment was repeated thrice, and one-step growth curves were plotted using GraphPad Prism 8  software. The burst size of the phage is equal to the amount of phage at the end of lysis divided by the initial number of bacterial cells at the time of infection  for 1 h. Three parallel controls were set up for each temperature, and titers were measured using the double-layer agar plate method. Additionally, the PaVOA phage was exposed to sterile Petri dishes at a distance of 40 cm from UV light (20 W) for 1 h, and samples were taken at 5 and 10 min intervals to determine phage stability. The titers of PaVOA phages were determined using the double-layer agar plate method at a pH range from 1 to 14, with pH conditions controlled using SM buffer. Experiments were repeated thrice.Host bacteria (PA/18) were inoculated in LB, LB-Ca, LB-Mg, and LB-Ca-Mg liquid medium  and then cultured in a shaking table at 37°C, 200 rpm until the OD600 was approximately 0.3. PaVOA phages were added at the optimal MOI as previously determined and cultured for 3 h. The resulting supernatants were collected by centrifugation at 12,000 rpm for 15 min at 4°C, with phage titers being determined by the double-layer agar plate method.9 PFU/mL with normal saline. New Zealand rabbits underwent shaving and disinfection of their ears, followed by slow injections of 1 ml diluted phage preparations through the ear marginal vein. Volumes of 500 μl whole blood were collected into anticoagulation tubes at different points  throughout the experiment. Whole blood samples were centrifuged at 12,000 rpm for 10 min at 4°C with the resulting supernatants being diluted in PBS. Phage titers at each time point were determined by the double-layer agar plate method to evaluate phage survival rate.New Zealand rabbits (3 months old) were purchased from Guangdong Provincial Medical Laboratory Animal Center . Purified PaVOA phage samples were diluted to a concentration of 10P. aeruginosa. Twenty-five New Zealand rabbits (3 months old) weighing approximately 1.9–2.1 kg were housed in sterilized cages and provided with adequate water and rabbit food. After 7 days of adaptive feeding, New Zealand rabbits were depilated at approximately 2 cm on the left side of the spine of their back, followed by local anesthesia using 0.5% lidocaine. Surgical scissors were then used to create square allograft wounds with a side length of approximately 2 cm  by syringe, after which wounds were covered with sterile dressings and adhesive bandages. Successful replication of the previously described model should result in purulent secretion on the wound surface, mild tissue erythema and edema, and slightly elevated skin temperature.The efficacy of phages in the treatment of traumatic skin bacterial infection was evaluated using phages and antibiotics to treat full-thickness skin excision wounds of New Zealand rabbits inoculated with ely 2 cm . Wound i8 PFU/mL), bacteriophage cocktail , and ceftriaxone groups (0.002%) received 2 ml of relevant solutions onto their sterile gauzes. Sterile adhesive dressings were then used to cover and fix all gauzes, followed by 4 days of treatment. Wound healing effects were evaluated at fixed points  using a digital camera and Vernier Calipers. Wound healing (%) was calculated using the following equation:Following the successful replication of the wound infection model, New Zealand rabbits were randomly divided into five groups: negative control (infection-free), positive control (infection), and three treatment groups , with five animals in each group. The administration methods for each group were as follows: negative and positive control groups were treated with 0.9% NaCl dropped onto their sterile gauzes, whilst single bacteriophage .All experimental data were collected and organized using Microsoft Excel and then plotted and analyzed using GraphPad Prism 8. Data are presented as mean ± standard deviation (SD).P. aeruginosa showed that all strains were highly resistant to cotrimoxazole, amoxicillin, and lincomycin. Additionally, more than 75% of the strains were highly sensitive to amikacin, ceftriaxone, cefepime, meropenem, ofloxacin, gentamicin, imipenem, azithromycin, tetracycline, ceftazidime, mezlocillin, cefoperazone, and tobramycin. The remaining 25% showed no obvious regularity concerning antibiotic resistance. Additional data showed that the host strain PA/18 was resistant to six antibiotics and was an MDR strain.The drug sensitivity results of the 22 antibiotics tested on 29 strains of Myoviridae family. Additionally, the host spectrum of PaVOA was determined by the spotting method, and the cleavage rate reached 8/29, indicating that the host spectrum of PaVOA was narrow.TEM showed that each PaVOA virus particle had an icosahedral head with a diameter of 66 ± 1 nm and a contracted tail with a length of 120 ± 7 nm , which iThe results of nuclease-mediated digestion of the phage genome revealed no bands in the DNase I treatment group, whilst the size and position of the bands in the negative control, RNase A, and the Mung Bean Nuclease treatment groups were effectively identical, indicating that the PaVOA genome is only sensitive to DNase I, thus proving that PaVOA possesses a double-stranded DNA (dsDNA) genome.An MOI of 0.1 resulted in the largest phage lysate titer . The resThe results of stability tests of PaVOA showed that the phage was able to tolerate low temperatures, and the survival rate did not change significantly between −20°C and 4°C. The highest survival rate was observed at 40°C. Decreasing survival was observed at temperatures above 40°C, and most notably at 70°C . The phap < 0.05). The addition of magnesium ions alone could improve the titer (p < 0.05), whilst the addition of calcium ions alone would reduce the titer (p < 0.05).The simultaneous addition of calcium and magnesium ions to growing PaVOA had the greatest effect on phage activity , which cAfter the phage was injected into rabbits, phage activity decreased rapidly within 12 h. As shown in The wound healing of the treatment and control groups on days 0, 3, 7, 12, 17, and 22 are shown in As shown in P. aeruginosa strains and produce plaques with a diameter of 2–3 mm. According to plaque morphology and TEM image identification, it is a lytic phage belonging to the Myoviridae family.Bacterial infections are a global public health issue , and treP. aeruginosa strains, whereas PaVOA only infected 27.6% of the tests strains. Additionally, phage PPaMa1/18 induced an 85.7% lysis of P. aeruginosa clinical isolates  that is resistant to six antibiotics to establish a New Zealand rabbit skin infection model for the comparison of phage (single or combined) and antibiotic (ceftriaxone) therapies for MDR wound infections. Overall, rabbits treated with ceftriaxone exhibited poorer wound healing than those treated with phages. Notably, treatment with a combined phage cocktail resulted in the greatest degree of wound healing. A previous study confirmed that phage cocktail treatment is effective against antibiotic-resistant Staphylococcus aureus diabetic foot ulcer infections  and Guangdong Province Modern Agricultural Industry Technology System, China (2019kj127).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.