It is demonstrated that the MRT boost in a 3 × 11 Gy radiation treatment has to be delivered as the last fraction to be more efficient for brain tumor control. p < 0.0001). The most efficient radiation regimen was obtained when the MRT-boost was applied as the last fraction, following two conventional clinical exposures.Synchrotron microbeam radiation therapy (MRT) is based on the spatial fractionation of the incident synchrotron beam into arrays of parallel microbeams, typically a few tens of micrometres wide and depositing several hundred Gray. This high dose, high dose rate, spatially fractionated radiotherapy has a high therapeutic impact on tumors, especially in intracranial locations. MRT leads to better control of incurable high-grade glioma than from homogeneous radiotherapy. The schedule of MRT within a conventional irradiation protocol (three fractions of 11 Gy) of brain tumors was evaluated on the 9L glioma model in rats. MRT delivered as a first fraction increased the median survival time of the animals by four days compared with conventional radiotherapy, while the last MRT fraction improved the lifespan by 148% (+15.5 days compared with conventional radiotherapy,  Our results highlight that one particular schedule is significantly favorable for improving the MRT-boost efficiency.2.All operative procedures relating to animal care strictly conformed to the guidelines of the French Government  and were conducted at European Synchrotron Radiation Facility (ESRF) in France.−1) for maintenance during tumor implantation and irradiations.The anesthetic procedure used in this study was isoflurane (5% in air) for induction and an intraperitoneal injection of xylazine/ketamine  of 8–10 weeks were implanted with 104 9L gliosarcoma cells in the right caudate nucleus as described previously  at a depth of 5.5 mm from the dura. Among rats bearing high-grade orthotopic tumors, animal groups were chosen on the basis of MRI T2W  in order to distribute the tumors homogeneously size-wise within the following groups of irradiation: 1 – group of untreated animals (n = 6); 2 – ConvRT only (three fractions of 11 Gy) (n = 10); 3 – ConvRT/ConvRT/MRT: 2 ConvRT fractions of 11 Gy + 1 MRT fraction (n = 10); 4 – MRT/ConvRT/ConvRT: 1 fraction of MRT + 2 fractions of 11 Gy ConvRT (n = 10); and 5 – ConvRT/MRT/ConvRT: 1 fraction of 11 Gy in ConvRT + 1 fraction of MRT + 1 fraction of 11 Gy in ConvRT (n = 9).Fisher 344 rats  from day 10 after implantation, either by ConvRT or MRT according to the group sorting.4.1.−1 was measured using an ionization chamber  placed in solid water plates  at 1 cm depth  and at room temperature. The ionization chamber and solid water plates were fixed at the same position and distance from the irradiation source as the rat head. A dose of 11 Gy was delivered to treated rats from day 10 after tumor implantation, depending on the animal group.Anesthetized rats were irradiated individually using orthovoltage photons performed on a Philips X-ray generator operated at 200 kVp, located on the ID17 beamline of the ESRF. Their sagittal suture was positioned on the left side edge of a 1 cm × 1 cm field for dorso-ventral irradiation of the entire rat’s right brain hemisphere (tumor volume/treatment volume ratio from 0.03 to 0.06). A dose rate of ∼1.4 Gy min4.2.−1. The beam was shaped into an array of thin, quasi-parallel microbeams using a multislit collimator . The wiggler produces a wide spectrum of photons  at a dose rate of ∼16000 Gy s al. 2012]. Fifty 5.GraphPad Prism version 9.0.1 .The survival curves were established using six rats for control, ten for ConvRT, ten for MRT/ConvRT/ConvRT, nine for ConvRT/MRT/ConvRT and ten for ConvRT/ConvRT/MRT groups. The time between the first fraction irradiation and death was recorded as the survival time; one day was added in cases of euthanasia. Kaplan Meier survival data and median survival time (MST) were compared using a log rank test in T/C × 100) – 100, where T and C are the mean survival days of treated and control groups of rats, respectively.The average increase in life span (% ILS) was calculated using the following formula: % ILS = .Kaplan Meier survival curves are shown in Fig. 26.2.p < 0.0001). MRT-boost as the last fraction appeared as the most efficient irradiation schedule used in this study ’. Fractions of MRT applied after several clinical irradiations (ConvRT) led to an increase in lifespan of glioblastoma-bearing rats (F98 tumor) compared with a total treatment with clinical irradiation . Five daily fractions of 5 Gy of the 67% isodose (maximum dose in the planning target volume of 37.5 Gy) yielded a significantly longer progression free survival for recurrent glioblastomas (from four months without boost to nine months with boost)  induced side effects and neurotoxicity. It will be followed by an MRT-boost trial to demonstrate MRT relevance in glioblastoma management. In early 2021, the Grenoble University Hospital gathered a ‘local task force’ which launched the procedures necessary for the first human clinical trial of MRT for glioblastomas. The benefit of dose escalation in the treatment of glioblastoma remains controversial but recent meta-analysis highlights a survival gain in patients (Singh  al. 2020 improved al. 2020 and Serd al. 2009, needs t al. 2021 demonstrTo conclude, this straightforward study demonstrates that MRT-boost improves the overall efficacy of ConvRT at controlling malignant gliomas and should be prescribed as a sequential boost following conventional fractions. To improve the MRT boost efficiency, we recommend to deliver the total radiation dose through a maximum number of ports, and the relevance of temporal fractionation in between ports needs to be further studied and demonstrated.


Vibrio parahaemolyticus is a foodborne pathogen diffusely distributed in the marine environment and often isolated from raw seafood belonging to different species, mostly shellfish. Ingestion of under- or uncooked seafood contaminated by V. parahaemolyticus can cause severe gastrointestinal symptoms in humans. Due to its ability to withstand low temperatures, Vibrio spp. could survive in frozen seafoods for long periods by entering the viable but nonculturable state (VBNC) and may constitute an unrecognized source of food contamination and infection. In the present study, seventy-seven frozen bivalve molluscs  were subjected to the detection and enumeration of viable V. parahaemolyticus using standard culture methods. VBNC forms were detected and quantified by applying an optimized protocol based on Propidium Monoazide (PMA) and Quantitative PCR (qPCR). All samples were negative for both the detection and enumeration of V. parahaemolyticus by the standard culture methods. VBNC forms were detected in 11.7% of the samples (9/77), with values ranging from 1.67 to 2.29 Log CFU/g. Only clam samples were positive for the detection of VBNC forms. The results of this study highlighted that VBNC V. parahaemolyticus may be present in frozen bivalve molluscs. Further data on the prevalence of VBNC V. parahaemolyticus in frozen seafood are needed in order to perform a robust risk assessment. Vibrio parahaemolyticus is a Gram-negative foodborne pathogen naturally present in marine and estuarine environments throughout the world. As a typical pathogen in seafood, it is one of the safety issues concerning these products, and food poisoning outbreaks caused by Vibrio spp. have been reported worldwide  and three fishing areas . Mussels included n. 32/35 (91.4%) shelled, n. 2/35 (5.7%) half-shell, and n. 1/35 (2.9%) whole-shell products all belonging to one species (Mytilus chilensis) and from the same fishing area (FAO zone 87). Samples were transferred to the laboratory under cold conditions and analysed within 24 h. One hundred grams of each sample were first thawed (<5 °C for 4 h) to avoid heat shock, then treated in a water bath at <20 °C for 15–20 min, homogenized, and divided into aliquots of 25 g. Each aliquot was diluted with sterile alkaline saline peptone water (ASPW)  in a ratio of 1:9 (w/v) and homogenized through a stomacher  for 2 min. Samples thus prepared were processed to determine V. parahaemolyticus via standard culture methods and PMA q-PCR to discriminate VBNC cells.Clam samples included n. 35/42 (83.3%) shelled and n. 7/42 (16.7%) whole-shell products belonging to five species  and two fishing areas .The nine VBNC positive samples belonged to the category of shelled clams and included three species [V. parahaemolyticus cells employing PMA associated with several molecular methods [V. parahaemolyticus, including tdh, trh, toxR, 16s rDNA, pR72H, gyrB, and vp1332 [tlh) encoding a phospholipase A2 is considered a species-specific marker for V. parahaemolyticus and is often utilised to identify this species [tlh gene was selected as the genetic marker in this study. The sensitivity of the PMA-qPCR assay was tested using serial dilutions of V. parahaemolyticus genomic DNA template. The LOD of PMA-qPCR was calculated to be 1.30 log CFU/mL, with the benefit of rapidity and specificity versus the standard culture method. Other qPCR assays developed in previous studies reported LODs for V. parahemolyticus of 1.2 × 102 CFU/mL [1 CFU/g [2O-labeled Raman spectroscopy, matrix-assisted-laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), liquid chromatography–mass spectrometry, autoradiography, ATP generation, and DNase I protection assay [In recent years, researchers have focused on the study of VBNC forms due to the wide variety of foodborne pathogens that can enter this state. VBNC cells, in fact, could pose a serious threat to food safety, given their ability to escape from traditional detection and enumeration methods, to withstand harsh situations , and to resuscitate and potentially lead to disease in consumers ,40,46. T methods ,65,66,67d vp1332 ,85,86,87 species ,88. Ther2 CFU/mL , 5 × 101[1 CFU/g , 12 CFU/[1 CFU/g . Previou[1 CFU/g . As repo[1 CFU/g ,76,89,90[1 CFU/g ,64. As r[1 CFU/g , the tim[1 CFU/g . In our on assay ,44. HoweV. parahaemolyticus was frequently responsible for causing disease, becoming a big threat for public safety [V. parahaemolyticus to tolerate low temperatures, and its detection in frozen products, extends the range of dangerous foods for consumers including also frozen seafood [V. parahaemolyticus during freezing and thawing, such as the extracellular concentration of solutes in the food, growth phase of the microorganism, presence of natural cryoprotectants , pH, temperature, freezing rate, exposure to chemical stresses before freezing, amount of nutrients in the environment, cooling or freezing medium composition, rate of cooling, dissolution, thawing speed, etc. [Vibrio is able to adapt to low temperatures through modifications of the composition of the fatty acids of the cell membrane and protein synthesis [Vibrio spp. can enter the VBNC state during freezing. A recent study [V. parahaemolyticus persisted in sea bass stored at 4 °C, −18 °C and −45 °C for 14 days; the fastest decrease in culturability, and a higher level of transition to the VBNC state were observed in samples stored at sub-zero temperatures. This is worrying, considering that studies have demonstrated how VBNC forms of Vibrio can become active again after cold storage and freezing due to temperature upshifts [Vibrio in products during thawing and, if the foods are not subsequently cooked adequately, cells that have regained their viable state could persist and cause disease. During the last decades, diseases triggered by foodborne pathogens have become a significant public health issue worldwide. Among these, c safety . In part seafood ,92. Seveed, etc. . Studiesed, etc. ,24,25 shnt study  showed tupshifts ,95. ThisV. parahaemolyticus with culture-dependent methods, while PMA-qPCR detected and quantified VBNC V. parahaemolyticus in 11.7% of culture-negative samples. Only clam samples were positive for VBNC V. parahaemolyticus. While there are numerous studies on the use of PMA-qPCR on artificially contaminated seafood samples, including shrimp, crab, fish, and shellfish [V. parahaemolyticus in marketed seafood. Yu et al. [In this study, all samples were negative for viable hellfish ,64,66,96V. parahaemolyticus in raw shrimp (Penaeus vannamei) and clams (Ruditapes philippinarum) purchased from various regions of Shanghai, did not find V. parahaemolyticus in any sample. In a study conducted by Zhu et al. [tdh-positive viable cells of V. parahaemolyticus in raw seafood, including oyster, scallop, shrimp, and crab, viable V. parahaemolyticus were isolated from three (10%) oyster samples, two (6.7%) shrimp samples, two (6.7%) crab samples, and one (3.3%) scallop sample. Recently, some studies on a new DNA-intercalating agent, PMAxx, produced on the basis of PMA, would have shown greater activity and better ability to distinguish between live and dead bacteria [Niu et al. , in a stu et al.  to invesbacteria . Howeverbacteria ,67,99. V. parahaemolyticus in frozen products. In fact, viable Vibrio spp., including V. parahaemolyticus, are commonly isolated from fresh bivalve shellfish worldwide [V. parahaemolyticus in 3.24% of samples . Tang et al. [V. parahaemolyticus were obtained by Lopatek et al. [Vibrio spp. in clams (Ruditapes decussatus) than in mussels  collected in Sardinia . According to these authors, the detection of V. parahaemolyticus only in clams in our study could be linked to the harvesting method. Clams, in fact, are usually harvested in muddy sand sediments while mussels are collected in ropes suspended in the water column. Since it has been demonstrated that loads of Vibrio spp. are higher in the sediments compared to the water column, the detection of VBNC V. parahaemolyticus only in clam samples in our study appears justifiable [V. parahaemolyticus were reduced to non-detectable levels in shucked oysters after 60 days (<3.8 log MPN/g) and to 0.38 log MPN/g in shell stock oysters after 75 days. These data suggested that frozen storage is an effective method for reducing V. parahaemolyticus, but also that Vibrio levels were higher in shelled than in shucked oysters. We can hypothesize that a preliminary handling of molluscs during the shelling process could result in higher initial bacteria loads and, consequently, more VBNC forms during freezing. Another factor to consider is seasonality. In this regard, a study [Vibrio were eliminated by oysters after depuration, while naturally present Vibrio persisted. This did not explain, however, why oysters that initially seemed characterized by low levels of Vibrio suddenly contained several logs more of such bacteria after exposure to a different bacterial population. This may be the consequence of a natural bacterial population already existing in a viable but non culturable state during cold months. In addition, it may be possible that the bacterium enters this form to survive the decreased temperatures. Several studies reported the absence of viable Vibrio in oysters during cold-water months, and their re-appearance when warmer waters return [Our results are substantially in accordance with other studies on viable orldwide ,101,102 orldwide , on 81 sg et al. , in frozk et al.   causing enterohemorrhagic and enteroaggregative diseases was discussed by Aurass et al. [Vibrio in frozen products are needed. Additionally, studies about the presence of VBNC Vibrio in seafood that are preserved differently, such as in increasingly popular ethnic seafood [Vibrio spp., including V. parahaemolyticus, were isolated from dried seafood [V. parahaemolyticus are resistant not only to cold temperatures but also to other stressors, such as heat, acidity, and low salinity [The results of this study showed that VBNC forms of in Japan . Asakurain Japan  have inss et al. . Even ifs et al. , 20% of  seafood , are req seafood ,30,115 aV. parahaemolyticus in commercial frozen bivalve molluscs. The positive results detected only for clams are probably linked to several factors, such as the harvesting method, initial contamination of the product during shelling operations, and seasonality. Our results show that PMA-qPCR can be considered as a fast and reliable method for the detection of VBNC pathogenic bacteria in food or during food processing, representing a useful tool for a more realistic risk assessment of V. parahaemolyticus in seafood products. Further studies are needed to acquire data about the prevalence of viable but non culturable V. parahaemolyticus in frozen bivalve molluscs and other seafood stored by cold or other preserving methods that could facilitate the development of VBNC forms.The detection of VBNC bacteria appears crucial to avoid false-positive or false-negative results at the industrial food production level. Underestimating the product safety profile can lead to serious consequences, since the occurrence of pathogenic species in food can result in serious outbreaks, and the potential re-growth of VBNC microorganisms could lead to a reduction in shelf life. Although the presence of VBNC bacteria is well reported, to our knowledge, this is the first study to report the presence of VBNC


Doxorubicin (DOX) is a frequent chemotherapeutic drug used to treat various malignant tumors. One of the key factors that diminish its therapeutic importance is DOX-induced nephrotoxicity. The first-line oral antidiabetic drug is metformin (Met), which also has antioxidant properties. The purpose of our study was to investigate the underlying molecular mechanisms for the potential protective effects of Met on DOX-triggered nephrotoxicity. Four animal groups were assigned as follows; animals received vehicle (control group), 200 mg/kg Met (Met group), DOX 15 mg/kg DOX (DOX group), and a combination of DOX and Met (DOX/Met group). Our results demonstrated that DOX administration caused marked histological alterations of widespread inflammation and tubular degeneration. Notably, the DOX-induced dramatic up-regulation of the nuclear factor-kappa B/P65 (NF-κB/P65), microtubule-associated protein light chain 3B (LC3B), neutrophil gelatinase-associated lipocalin (NGAL), interleukin-1beta (IL-1β), 8-hydroxy-2′ -deoxyguanosine (8-OHdG), and Beclin-1 in renal tissue. A marked increase in the malondialdehyde (MDA) tissue level and a decrease in the total antioxidant capacity (TAC) were also recorded in DOX-exposed animals. Interestingly, Met could minimize all histopathological changes as well as the disruptions caused by DOX in the aforementioned measures. Thus, Met provided a workable method for suppressing the nephrotoxicity that occurred during the DOX regimen via the deactivation of the Beclin-1/LC3B pathway. Chronic kidney disease (CKD) is a set of chronic progressive nephropathy associated with a steady loss of renal function. Recently, the prevalence of CKD has risen to reach 14%, and it coincided with an increased mortality rate . Renal iDoxorubicin (DOX) is one of the most competent anthracycline drugs against different cancer types. However, its limited efficacy was related to systemic toxicity, particularly kidney injury . It is aAutophagy is a lysosomal degradation mechanism for clearing and reusing cytoplasmic components . AutophaMetformin (Met), a metabolic regulator, is extensively used with type-II diabetics . Met effThus, this study was assigned to assess the possible protective effect of Met against nephrotoxicity inflicted during the DOX regimen and to unveil its underlying antioxidant, anti-inflammatory and anti-autophagy mechanisms involving Beclin-1 and LC3B. p < 0.001). Meanwhile, a significant decrease in weight was observed in the Met prophylactic group when compared to the DOX group (p < 0.001). The co-therapy group significantly reduced kidney weight by 23.07% . The combination of DOX and Met markedly reduced the renal NGAL content by 52.8% in comparison with the DOX group (p < 0.001).In The DOX group showed a marked increase in the MDA renal content by 58.38% in comparison with the control group. While diseased animals treated with Met resulted in a significant decrease in MDA renal content by 48.07% compared to the DOX group A. Followp < 0.001). The Met prophylactic effect reduced renal 8-OHdG content. The prophylactic group displayed a significant reduction in renal 8-OHdG by 42.41% compared to the DOX group  within the renal tubular lining epithelium C. The kiInterleukin (IL)-1β is known as a signaling proinflammatory biomarker. In comparison with the normal group, DOX treatment caused a considerable rise in renal IL-1β level of 80.11%. On the other hand, Met treatment caused a significant reduction in renal IL-1β concentration. When compared to the DOX group, prophylactic Met  administration showed a substantial decrease in renal IL-1β level by 60.54% F.The control and Met groups exhibited negative mild cytoplasmic immunostaining for renal LC3B C. HoweveBeclin-1 protein expression was significantly elevated by approximately 60%, 57.24% in the DOX-treated mice compared with the control and Met groups. Prophylactic treatment with Met reduced the renal Beclin-1 protein expression by 24.79%, compared with the DOX group F.The clustering heatmap depicted in Next, to determine the most influencing parameter in response to different treatments in the current study, VIP score was conducted. As depicted in DOX is one of the most potent anthracyclines for treating various cancers. However, the DOX regimen is frequently linked to nephrotoxicity . DespiteExpectedly, DOX could exert renal damage indicated by elevated levels of serum creatinine and urea. A dramatic increase in the NGAL was also noticed in DOX-treated animals. It is well known that NGAL excretion in the urine is a recognized indicator for tubular epithelial injury. Along with these, the current histopathological examination exhibited the existence of tubular degeneration and loss of tubular epithelium which was the reason that contributed to the elevated creatinine, urea, and NGAL levels.2•−), hydrogen peroxide (H2O2), hydroxyl radical (OH•), and nitric oxide (NO) into the cytosol can cause further damage to other cellular macromolecules including disintegration of lipid bilayer membranes, mitochondrial dysfunction, protein misfolding, endoplasmic reticulum (ER) stress, and DNA damage [• can distantly target the cellular molecules, specifically the lipid content of cell membranes, leading to lipid peroxidation and the subsequent generation of another damaging compound . Consistently, our data revealed a marked increase in MDA levels along with depletion in the TAC; thereby, the cell becomes more vulnerable to the damaging effect of ROS and MDA. Therefore, the noticed tubular degeneration and loss of tubular epithelium might be attributed to the generalized damage brought on by the buildup of the damaging radical (ROS and MDA). Moreover, oxidative species have been implicated in disrupting the nephron’s excretory function, which leads to an imbalance in homeostasis and the accumulation of metabolic waste products. It is widely known that renal cells include a large number of mitochondria, providing the adenosine triphosphate (ATP) needed for active transport and the removal of metabolic byproducts. In the meantime, those harmful radicals inhibit the mitochondrial oxidative phosphorylation required for ATP synthesis. We therefore strongly suspect that the observed renal impairment, as indicated by increased levels of creatinine, urea, and NGAL, was caused by oxidative damage brought on by DOX.It has been hypothesized that the primary process driving the nephrotoxic impact of DOX is oxidative stress and its downstream events such as free radical overproduction, lipid peroxidation, and antioxidant enzymes depletion ,23. OxidIn the current investigation, DOX administration substantially enhanced the NF-κB/P65 and IL-1β expression levels in the renal tissues which is consistent with the previously published data . It is iIt is well known that the autophagy process, including the removal of protein aggregates and damaged organelles, improving cellular energy production, and reduction the endoplasmic reticulum stress, is required for maintaining cell physiology and survival. Thus, for the preservation of kidney function, basal autophagy appears to be crucial for the survival of podocytes, proximal tubular epithelial cells, glomerular mesangial cells, and glomerular endothelial cells . As prevFurthermore, Beclin-1 is also a critical determinant of whether cells act in autophagy . Beclin-Hence, Met has antioxidant and anti-inflammatory activities, and it could significantly restore kidney function in DOX-treated animals shown in the measured creatinine, urea, and NGAL. Met has also an additive advantage through reduction of the degree of insulin resistance, improvement of the renal proximal tubular glomerular filtration function, and lowering the blood glucose . Met treRemarkably, Met protects young C57BL/6 mice fed a high-fat diet from obesity-induced kidney damage . The undIn addition, DOX-exposed animals treated with Met showed a significant decrease in the levels of Beclin-1 and LC3B. These findings are consistent with the role of Met in the modulation of different signaling pathways, including the adenosine monophosphate-activated protein kinase (AMPK) pathway, the Hedgehog pathway, and microRNAs and autophagy block through miR-570-3p, miR-142-3p, and miR-3127-5p . The AMPThe clustering heatmap provides an intuitive visualization of the concentration values that could potentially discriminate the DOX-treated group from other treatments. Wherein, the DOX showed the occurrence of severe alterations was confirmed by the great differences shown in the concentration values of all variables when compared to the control and Met-treated animals. However, the supplementation of Met to the DOX-injured animals exhibited markable improvements exhibited by the intermediate intensity of the concentration value of all variables as depicted in ® vial, each ml contains 2 mg) was purchased from EIMC United Pharmaceuticals Company . Met powder was purchased from Sigma-Aldrich . DOX  were obtained from the laboratory animal house of the Modern Veterinary Office Faculty of Veterinary Medicine, Cairo University, Cairo, Egypt. Animals were kept in well-acclimatized environmental conditions of a 12 h light and dark cyclic conditions and at a temperature of 25 ± 1 °C. Throughout the study, both food and water were accessed ad-libitum. The research protocol was following the Canadian Council on Animal Care Guidelines and was approved by the committee of Research Ethics, Faculty of Veterinary Medicine, Kafrelsheikh University (KFS/12/21).2. The fourth group was administrated a combination of DOX and Met. Treatment with Met was started one week ahead of the DOX administration and continued for 14 days as described in Fourteen days later, all animals were randomly categorized into four groups (five mice per each group). The first group acted as a control group; received only saline. The second group received a daily dose of Met  . A singlTwenty-four h after the last drug administration, all animals were euthanized under deep anesthesia via thiopental sodium at a dose of 50 mg/kg. A blood sample was obtained from the abdominal aorta, allowed to be clotted for 10 min and then sera were separated after centrifugation for 10 min at 3000 rpm. After that, animals were dissected, kidneys were harvested and then one kidney was fixed in 10% paraformaldehyde solution for histopathological and immunohistochemical investigation. The remaining kidney was frozen at −80 °C for ELISA assays.According to the commercially available kits, serum creatinine and urea were assessed in accordance with their manufacturer’s instructions .Assay of NGAL was performed using the manufacturer’s instructions provided by the commercial available ELISA kit .The oxidative stress parameters including MDA , TAC , and 8-OHdG  contents were measured in the renal homogenate using the available commercial ELISA assay kits. Test procedures were carried out according to the provided manufacturer’s instructions.Beclin-1was determined using the commercially available ELISA assay kits . The renal tissue content of IL-1β was assessed using a Mouse IL-1β ELISA kit .The fixed tissue samples were rinsed down in tap water for 10 min, dehydrated in ascending grades of ethyl alcohol, and cleared in xylene. After that, they were immersed in molten paraffin at 58–62 °C. About 5 µm histological sections were made and stained with hematoxylin and eosin. The slides were ready to examine under a bright-field microscope. 2O2 solution for the deactivation of endogenous peroxides followed by protein blocking. Then, sections were incubated with an anti-LC3B antibody, an autophagosome marker , and anti-NF-ĸB/P65  at a dilution rate of 1:100. The slides were then incubated with secondary goat anti-rabbit antibody  for polyclonal antibodies for 30 min at room temperature following phosphate buffered saline rinse. Slides were then covered with a drop of DAB kit solution and eventually counterstained with Mayer’s hematoxylin. The staining labeling indices of the antibodies were presented as the mean of the percent of positive area/mm2.The immunohistochemical staining technique was performed by Khalil et al. . Sectiont-test and Mann–Whitney U test for two-group statistical comparison of parametric and nonparametric data, respectively. p < 0.001 was accepted as being statistically significant. Data were plotted by the OriginPro software (version 2019b). RStudio (R version 4.0.2) was used to create the clustering heatmap and the variable importance in projection (VIP) score. The “pheatmap, dendextend, gplots, colorspace, vip” packages were used to generate those analyses.The numerical data were expressed as mean ± SEM. Statistical analysis was conducted using Student’s The present research demonstrated the possible renal protection against DOX-inflicted injury through oral Met treatment. Marked improvements in kidney function by Met administration via reduction of oxidative stress, suppression of pro-inflammatory cytokines, and reduction of DOX-induced autophagy were documented. The therapeutic potential of Met was possibly attributed mainly to its antioxidant and anti-inflammatory properties.


The growing demand for watertight concrete structures is conducive to the development of research in this area, but their results are rarely published. In order to partially fill this gap, the authors of the publication present the results of research into the effect of fly ash addition on the watertightness of concrete. Prior to the tests, a recipe for a concrete mix with the addition of a sealing admixture modified with fly ash was developed. The following properties were analyzed: consistency of the concrete mix, air content in the concrete mix, compressive strength of concrete, depth of penetration of water under pressure, and frost resistance of concrete for F150 level. The work meets the expectations of the construction industry with respect to the production of concrete structures resistant not only to the penetration of water into concrete but also resistant to aggressive substances dissolved in water that accelerate the destruction of concrete and corrosion of reinforcement bars. Based on the test results, it was found that the addition of fly ash to the concrete mix enhances the positive impact of the applied sealing admixture, increasing the tightness of the concrete. It reduces the depth of penetration of water under pressure and therefore increases the frost resistance of concrete. Although concrete is one of the oldest building materials, it is still subjected to various modifications in order to improve its physical and mechanical properties and durability in an aggressive environment. Currently, one of the basic components of concrete is modern chemical admixtures, thanks to which it is possible to optimize its properties. The variety of available chemical admixtures allows for their selection in terms of improving both the properties of the concrete mix and the durability of concrete . The rapid development of chemistry also enables the use of various additives, which are materials usually used in quantities exceeding 5% of the mass of cement. They are divided into two types; one is inert additives in the form of filler of aggregates or pigments, and the other type has pozzolanic properties  or hydraulic properties (granulated blast furnace slag). The use of admixtures and additives allows modification of the concrete mix and/or hardened concrete. Concern for the environment, in accordance with the principles of sustainable development, makes it necessary to use ecological construction products that allow reducing the consumption of cement, the production of which has a negative impact on the environment, and natural construction aggregates, whose easy to extract resources, are slowly running out. In Jura et al. , the posIn recent years, the demand for building structures made of watertight concrete has been increasing rapidly. The limited availability of building lots in large cities means that many newly erected buildings have underground parking floors, which, due to the environment and harmful external conditions, need complete protection against water penetration into their structure. In addition, the intensive development of civilization causes the formation of “urban heat islands” with a much higher temperature compared to their outskirts. In order to counteract this, more and more buildings are erected with roofs covered with vegetation, which act as natural heat accumulators . AccordiMousavi et al.  showed tHowever, it should be remembered that the “green roof” technology requires 100% protection of the ceiling against moisture penetration. The residents of the famous Villa Savoye, designed in 1928 by the French architect Le Corbusier, learned painfully about the consequences of not meeting this condition. After a few months after its erection, they had to move out of the building, as it turned out that water leaked through the roof, and the repair process turned out to be unprofitable and difficult to carry out . CertainThe impact of aggressive groundwater causes corrosion and damage to concrete structures as a result of corrosion of steel reinforcement and concrete cracks. The durability of concrete structures largely depends on the resistance to water penetration and chlorine, sulfur, or carbon dioxide compounds dissolved in it. According to , the proCurrently, we are dealing with increasingly larger and deeper underground structures that are subject to destruction to a much greater extent than before. Unfortunately, classic concrete does not meet the waterproofing criteria. The thermal effects of cement hydration cause thermal stresses, which in turn cause the formation of a network of microscopic channels through which water easily migrates into the concrete. This is especially dangerous in the winter, when the freezing water increases in volume and bursts the concrete. Water leakage in concrete elements poses a significant issue as it shortens the structure’s lifespan and necessitates continuous maintenance. According to , factors2-CH3 nanoparticles dispersed in vegetable oil, used to facilitate formwork demoulding, despite the small penetration depth, improves the water resistance of concrete without affecting its compressive strength. The performance of the moisture barrier system can be significantly enhanced by the use of waterproofing/dampproofing admixtures in concrete [One of the possibilities to improve the durability and reliability of concrete structures and to increase the comfort of their use is the application of modern materials and construction technologies. Until recently, the main method of extending the durability of concrete structures was to use external insulation in the form of coatings, foils, membranes, or bituminous felts ,23,24,25concrete . They caconcrete , it was 2 sol and silane emulsion. They showed that the hydrophobic effect was obtained due to the morphological changes of the concrete (C40), mainly the improvement of the concrete surface microstructure. Muwashee et al. [At work , the impe et al.  used bene et al. , an inexe et al. ,37, see e et al.  showed ae et al. , in ordeStudies by Matar and Barhoun  on the iWhile watertight concretes, enhanced with specialized admixtures and additives, have been increasingly used in recent years and become impermeable to water after cur-ing, Chew and Silva  emphasizWaterproof concrete structures, the so-called “White Box Concept,” are designed and built to prevent water leakage from or into the structure. B. H. Cho et al.  conducteThe literature review indicates that, at the moment, the best solution to ensure the tightness of concrete structures is the use of watertight concrete in combination with an appropriate system for sealing expansion joints and all elements included in the structure. Despite the growing demand for waterproof concretes, there are few works on their design in the available technical literature, especially in open access. Companies protect their formulas and rarely share their knowledge in this area. Therefore, the authors of this work decided to present the design of the concrete mix as well as the results of tests of the concretes intended for structures exposed to groundwater pressure, such as garages, basements, or other types of underground technical rooms. The main goal of the work is an analysis of the effect of flay ash addition on the watertightness of concrete. Our test results showed that fly ash enhanced the effect of the sealing admixture, additionally limiting the depth of penetration of water under pressure, which is extremely important not only in the case of concrete used for underground structures but also for the construction of green roofs, which is a kind of novelty on the market. In the absence of detailed guidelines for the design of watertight concretes the test results can be used by technologists and concrete designers as guidelines in the selection of the composition of watertight concrete. Experiments allow for reducing mistakes in the future.-V1: reference concrete mix with an admixture of plasticizer and superplasticizer,-V2: concrete mix of variant 1 modified with the addition of a sealing admixture,-V3: concrete mix of variant 1 modified with the addition of fly ash used as a replacement for a part of the aggregate,-V4: concrete mix of variant 3 modified with the addition of a sealing admixture.In order to study the effect of flay ash on the watertightness of concrete., the concrete mixes with the addition of sealing admixture, which was modified with fly ash, were developed. As a part of the work, the following 4 variants of concrete mixes for making watertight concrete were designed:The adopted designations of the mixes correspond to the order of design and execution of the tested series of concretes. First, a reference series (V1) was designed experimentally, and then, the reference series was modified with the addition of a sealing admixture and marked as V2. In the next stage, the reference series was redesigned by introducing fly ash (V3), and then a sealing admixture was added to the V3 series, and thus the V4 series was created. When designing concrete mixes, it was taken into account that the technology of water-tight concrete is effective only when we are able to eliminate possible cracks that arise not only as a result of construction errors but also improper technology of concrete production. Therefore, special attention was paid to the selection of concrete mix components, especially the selection of admixtures and additives to improve the plasticity and tightness of concrete and to ensure the proper ratio of water to cement and the optimal distribution of aggregate grains.-consistency of the concrete mix,-air content in the concrete mix,-concrete compressive strength,-depth of penetration of water under pressure,-frost resistance of concrete for level F150.Experimental studies of concrete mixes and hardened concrete included testing:Portland slag cement CEM II/B-S 42.5N NA  was usedBV3M plasticizer, ViscoCrete 5 superplasticizer, and WT-200P sealing admixture were used as concrete admixtures. Silica fly ash with pozzolanic characteristics meeting the requirements of the standard PN-EN 450-1:2012 [When developing the recipe for concrete mixes, the following parameters were sought: water penetration depth: <30 mm, degree of frost resistance: minimum F150, consistency class after 5 min from the first contact of water with cement: S2, air content in the concrete mix after 5 min from the first contact of water with cement: 2 ÷ 3%.As part of the design of the composition of the concrete mix of variant V1, gradation tests were carried out, and then the mixing proportions were experimentally determined in order to obtain the optimal graining of the aggregate by examining its bulk density and voids. The grading curve of the aggregate mix, together with the limit curves according to PN-B-06265:2022-08 , is showA sealing admixture in the amount of 1% of the cement mass was introduced into the designed composition of the concrete mix of variant V1 (variant V2). In variant V3 of the concrete mix, silica fly ash was introduced into the composition as a substitute for part of the aggregate without using a sealing admixture. As part of further modifications in variant V4, fly ash and a sealing admixture in the amount of 1% of the cement mass were used at the same time. The recipes of the tested concrete mixes are presented in Fly ash in V3 and V4 concrete mixes was introduced into the composition as a substitute for part of the aggregate. All designed concrete mixes meet the requirements of concrete durability in a chemically aggressive environment.In order to check whether the designed concrete mixes meet the design assumptions, qualifying the concrete for use in the construction of facilities exposed to groundwater, standard tests of the obtained concretes carried out, such as consistency of the concrete mix, air content in the concrete mix, concrete compressive strength and depth of water penetration under pressure. Due to the fact that concretes are also used in cold regions exposed to excessive freezing and thawing during the year, an additional frost resistance test of concrete was carried out. The purpose of such a test is to check the resistance of concrete to the destructive effects of changing temperatures. All tests were carried out in accordance with applicable standards.The test of the consistency of the concrete mix was carried out using the concrete slump test in accordance with the standard PN-EN 12350-2:2019-07 . The conIn addition to affecting the strength of concrete, the air content can also affect the migration of harmful compounds and the durability of concrete in various environmental conditions. It can also impact the finish and cosmetic appearance of concrete. Therefore, air content testing is required for concretes used for structures that may be exposed to the aggressive effects of the surrounding environment.The air content in the fresh concrete was tested using the pressure method . in accoThe compressive strength was tested on a hydraulic strength press  in accorThe concrete mix was placed in molds in two layers, each of which was compacted mechanically on a vibrating table. The samples remained in the forms for 24 h. After demoulding, they were placed in water, where they were cured at the temperature of 20 ± 2 °C. In the case of cubic samples with an edge of 100 mm, the compressive strength was tested after 2, 7, 28, 56, and 90 days, and in the case of the samples with an edge of 150 mm, the tests were carried out after 7, 28 and 90 days.The depth of penetration of water under pressure into concrete specimens was tested in accordance with the standard PN-EN 12390-8:2019-08 . The tesThe study was conducted continuously for 72 ± 2 h. After this time, the samples were broken , and theIn countries such as Poland, where subzero temperatures are common in winter, it is necessary to test concrete for frost resistance because water in concrete pores freezes under the influence of negative temperatures, increasing its volume by about 10%. This causes significant stresses, leading to cracking of the concrete and loss of its tightness. Cyclic repetition of this phenomenon can lead to the complete destruction of the concrete structure. Therefore, freeze-thaw tests were carried out in accordance with the standard PN-B-06265. The tests were carried out on cubic samples with an edge of 150 mm after 56 days. The samples were subjected to 150 cycles of freezing in the air to a temperature of −18 ± 2 °C within 4 h and thawing in water at a temperature of 18 ± 2 °C for the next 4 h. After the last cycle the samples were weighed and tested for compressive strength. According to the PN-B-06265 standard, the frost-resistant level of mature concrete is achieved when, after the cycles of freezing and thawing: (1) none of the tested concrete samples shows cracks, (2) the average weight loss of the tested samples does not exceed 5%, (3) compressive strength of samples subjected to freeze-thaw cycles is not lower than 20% compared to the control samples.The results of the consistency test are presented in The results of testing the consistency of concrete mixes indicate that for all variants of concrete mixes, the assumed consistency class S2 was obtained after 5 min. An increased drop of the cone of the tested mixes in the time interval of up to 30 min can be seen for all tested variants. The effective action of the fluidizing admixtures was obtained after 30 min from the moment of combining the cement with water in the mixing process. A slight decrease in consistency after 30 min should be explained by of hydration of the cement binder. The concrete mix, according to variant V4, with the addition of fly ash and sealing admixture, showed the greatest liquefaction. Between 30 and 60 min, all concrete mixes retained their workability, and the cone slump was 90 mm.The results of air content in the fresh concrete are presented in All concrete mix recipes met the assumption of air content within the limit of 2 ÷ 3%. Concrete mixes containing silica fly ash (variants V3 and V4) are characterized by a slightly higher air content. It can be seen that the addition of a sealing admixture reduces the air content in the tested mixes, both in the case of its introduction to the reference concrete mix (variant V1) and after its introduction to the concrete mix of variant V3 (with the addition of fly ash). In the concrete mix of the V1 variant, compared to the V2 variant, the air content decreased from 2.3% to 2.2%, whereas in the concrete mix of the V3 variant, compared to V4, there was a decrease from 2.6% to 2.4%.The test results of compressive strength are presented in The tests showed that after 28 days of concrete curing, the compressive strength class C50/60 was obtained for all four recipes, and the strengths were similar to each other. The largest difference occurred between variants V2 and V4 and amounted to about 4%. The concrete of variant V4, with the addition of fly ash and sealing admixture, obtained the highest value of compressive strength, which was 71 MPa. This may be due to the synergistic effect of the fly ash with the sealing admixture, which has an impact on the elimination of micro-cracks in concrete.The addition of fly ash reduced the compressive strength in the first days of concrete maturation (variants V3 and V4), but in the later period, concretes with the addition of fly ash achieve a greater increase in strength, specifically the concrete according to variant V4 obtained the highest compressive strength after 90 days of maturation.After 2 days of concrete curing, the highest strength of 28 MPa was obtained in the reference series, while in the variants with the addition of fly ash, the strengths were the lowest (variants V3 and V4). Between the 2nd and 7th day of maturation, concretes with the addition of fly ash achieved a higher increase in compressive strength compared to the series without ash, but the strengths were still slightly lower. After 28 days, very similar results of compressive strength were obtained in all variants, and the greatest increase in strength was obtained for the concrete variant V4. In the further period of concrete curing between 56 and 90 days, a further increase in compressive strength was observed for variants V3 and V4 containing fly ash.The results of water penetration, i.e., the depth at which water penetrated the concrete sample, are shown in The depth of water penetration into the concrete of the reference series (V1) was 41 mm. For the remaining modified concrete mix recipes (variants V2 ÷ V3), the assumed water tightness of concrete was obtained, i.e., the depth of penetration of water under pressure did not exceed the assumed limit of 30 mm. The analysis shows that the sealing admixture (variant V2) and fly ash (variant V3) affect the reduction in the penetration depth. Modification of the reference series of concrete (variant V1) by introducing fly ash into the concrete mix (variant V3) resulted in a reduction in water penetration by 39%, while modification of variant V1 with sealing admixture (variant V2) resulted in the reduction in water penetration by 56%. In the case of variant V4, the addition of fly ash enhanced the effect of sealing admixture, reducing water penetration by 61%. The synergy of both components effectively seals the concrete structure, creating a barrier that is hardly permeable to water.Such concrete reduces not only the water penetration into the concrete but also prevents the ingress of aggressive substances dissolved in water, which in turn decreases the destruction of concrete and corrosion of reinforcing bars.In all tested variants, no loss of mass for samples subjected to freezing and thawing cycles was recorded. The results of the strength decrease tests are shown in The analysis of the freeze-thaw tests showed that the assumed F150 class was obtained for all four concrete mix recipes. None of the tested samples was cracked or scratched. Concrete, according to variant V1, shows the greatest decrease in compressive strength, i.e., 5.46%. The smallest decrease in strength, i.e., 1.45%, was observed for concrete according to variant V4. The reason for the better results in the case of variant V4 is a more airtight structure compared to other concretes due to the use of both a sealing admixture and fly ash, which reduces the access of water to the interior of the concrete.The applied sealing admixture had a positive effect on the tightness of the concrete and thus improved the frost resistance of the concrete. A 56% decrease in water penetration from 41 mm for the V1 series to 18 mm for the V2 series was observed.Fly ash enhanced the effect of the sealing admixture, additionally limiting the depth of water penetration under pressure. For the V4 series, the depth of water penetration was 16 mm, which is a reduction in this parameter by 61%.The addition of fly ash to the concrete mix improved its workability and increased the air content in the concrete mix. It reduced the compressive strength in the first stage of maturation but increased it later, i.e., after 90 days.The sealing admixture has a positive effect on the tightness of the concrete, which is reflected in the compressive strength. After 90 days of curing, it was found that in concrete without fly ash, the strength increased by 1.1% (series V1 and V2), while in the presence of fly ash by 1.2% (series V2 and V3).Analysing the properties of the tested concretes, it can be concluded that the most favorable results were obtained for the V4 series, which contained the addition of fly ash and a sealing admixture.Waterproof concrete is a modern building material that allows for replacing traditional solutions using external insulation in the form of coatings or membranes. Appropriate design of the concrete mix and execution of the concrete ensures the extension of the service life of the concrete structures. Based on the obtained results of the conducted tests, it can be stated that:In the absence of detailed guidelines for the design of watertight concretes the test results can be used by technologists and concrete designers as guidelines in the selection of the composition of watertight concrete.Future research will include an analysis of the possibility of using fly ash from municipal waste incineration in the composition of watertight concrete.


The administration of clinically indicated antibiotics prior to surgery in combination with other evidence-based practices reduces the risk of surgical site infection (SSI). A retrospective study of SSI at a community-based hospital that performs 5,700 procedures annually revealed broad variability in the timing of prophylactic antibiotic administration in relation to the time of incision. A Tableau dashboard was created to compare antibiotic administration to surgical incision timing and the impact variability may have on the incidence of SSI.A multidisciplinary team was formed with surgeons, anesthesia, nursing, antimicrobial and diagnostic stewardship, infection prevention, and analytics representatives. Antibiotics selected for inclusion were those most prescribed for surgical prophylaxis according to organizational practice guidelines. Using the dashboard, optimal administration times were identified based upon the pharmacokinetics of the antibiotic as well as the expected tissue concentration at the incision site when available. (See table) Surgical encounters that received an antibiotic were classified as “pass”, “fail”, or “other”. Antibiotics started within the determined timeframe passed, those started earlier or later than the timeframe failed, and the others received antibiotics not included in the dashboard logic.In a baseline review 35.5% of cases failed timing criteria. Failures occurred most frequently with cefazolin, ceftriaxone and clindamycin. A retrospective analysis of SSI (n = 34) classified using the National Healthcare Safety Network (NHSN) definitions correlated with the baseline data. The outliers were comprised of antibiotics started too close to or after incision (14.7%) and antibiotics started much earlier than the dosing timeframe and incision (17.6%).The surgical prophylaxis timing dashboard helped to identify gaps in administration practices and highlighted the need for process improvement initiatives. Ongoing interventions to improve adherence to national guidelines will be evaluated to determine the impact on SSI rates.All Authors: No reported disclosures


However, understanding its tension stiffening and crack spacing and width is of critical importance in fulfilling serviceability requirements. Therefore, this research aims to evaluate the tension stiffening and cracking performance of alkali-activated (AA) concrete. The variables considered in this study were compressive strength (fc) and concrete cover-to-bar diameter (Cc/db) ratios. After casting the specimen, they were cured before testing at ambient curing conditions for 180 days to reduce the effects of concrete shrinkage and obtain more realistic cracking results. The results showed that both AA and OPC concrete prisms develop slightly similar axial cracking force and corresponding cracking strain, but OPC concrete prisms exhibited a brittle behavior, resulting in a sudden drop in the load–strain curves at the crack location. In contrast, AA concrete prisms developed more than one crack simultaneously, suggesting a more uniform tensile strength compared to OPC specimens. The tension-stiffening factor (β) of AA concrete exhibited better ductile behavior than OPC concrete due to the strain compatibility between concrete and steel even after crack ignition. It was also observed that increasing the confinement (Cc/db ratio) around the steel bar delays internal crack formation and enhances tension stiffening in AAC. Comparing the experimental crack spacing and width with the values predicted using OPC codes of practice, such as EC2 and ACI 224R, revealed that EC2 tends to underestimate the maximum crack width, while ACI 224R provided better predictions. Thus, models to predict crack spacing and width have been proposed accordingly.Alkali-activated concrete is an eco-friendly construction material that is used to preserve natural resources and promote sustainability in the construction industry. This emerging concrete consists of fine and coarse aggregates and fly ash that constitute the binder when mixed with alkaline activators, such as sodium hydroxide (NaOH) and sodium silicate (Na Concrete made with Ordinary Portland Cement (OPC) has been commonly used as a construction material for various types of structures for almost 200 years ,2. Nowad2SiO3), in a process called geopolymerization [Recently, there has been an increased motivation among concrete practitioners and researchers to explore the potential of using AA concrete as an eco-friendly alternative to OPC concrete. In addition to the fine and coarse aggregates, FA is a primary component in AA concrete that is produced through mixing all previous components with an alkaline activator, such as sodium hydroxide (NaOH) and sodium silicate  ratio on tension stiffening, crack spacing, and crack width and propose models for crack spacing and width based on the experimental results.The outcomes of the present study on tension stiffening and cracking performance are expected to help better understand the serviceability limit state behavior of AA concrete. This is because the tension-stiffening behavior plays a significant role in the width and spacing of cracks, stiffness, and deformation of the structural member; thus, it is often the governing design criterion for longer-span reinforced concrete members. Understanding tension stiffening also enables engineers to perform accurate deflection calculations for serviceability limit state analysis. This is also significant when dealing with deformability analysis, particularly when cracking is involved, as it is a major source of nonlinearity. Furthermore, relying on the existing models of OPC concrete to estimate the crack spacing and crack width of AA concrete could lead to an unsafe or suboptimal design of structural members; the developed crack spacing and crack width models in this study could provide a safer design basis for structural engineers. Therefore, this study aims to investigate the effects of compressive strength and concrete cover-to-bar diameter  and are presented in STM C618 . The OPC2SiO3) and 14M sodium hydroxide (NaOH). The percentage of Na2SiO3 solution to the 14M NaOH was maintained at 1.5 by weight for all mixes, as suggested by Abdulrahman et al. [2SiO3 and 12.72% NaOH by weight. It is worth mentioning that the three optimum concrete admixtures developed by Abdulrahman et al. [The alkaline activator solution employed in this study was a combination of premixed sodium silicate  was applied to the protruded part of the steel bar until its yielding limit. Although a standard test setup for concrete elements under tension does not exist, the direct tensile test is the most widely used experimental layout . The loaThe results of the scanning electron microscopic (SEM) analysis of the FA and OPC powder are given in 2), mullite (Al6Si2O13), magnetite (Fe3O4), and hematite (Fe2O3). The figure also shows that increasing the intensity occurs at angles between 27 and 44, where the components are high.The results of the X-ray diffraction (XRD) analysis of FA are given in cr) and the corresponding cracking strain (ɛcr) of all prisms, together with the tension force  and strain  of reinforcement, as well as the cracking force (Pcr) and stress (Fcr) of concrete. The characteristics compressive strengths (fc) of AA concrete vary between 24 and 43 MPa, while the characteristics compressive strengths of OPC concrete were 28 MPa and 35 MPa, respectively. Despite the slight difference in concrete strength for both AA and OPC specimens, the obtained results were used to compare the mechanisms of tension stiffening and crack response for both concretes.The experimental results of tension stiffening for AA prisms along with the OPC control prisms are summarized in cr), the corresponding cracking strain (ɛcr) of OPC prisms was slightly smaller, as shown in cr). This enabled the OPC concrete to exhibit higher tensile stresses (Pcr) before cracking, which suddenly deteriorated after the initial crack form. However, the tensile stress (Pcr) in AA concrete was slightly lower at the initial crack, but it continued to carry more tensile stresses after cracking, resulting in a better tension stiffening than OPC concrete. These premature cracks observed in the current study are similar to those reported in the literature for AA concrete [Although both AA and OPC prisms developed slightly similar axial cracking force . This stage occurred when crack patterns in at least two consecutive load stages remained constant, and only the crack width increased. Thus, the response of the cracked prisms started to approach that of the bare bar, and the maximum applied load was limited by the yield strength of the reinforcement. This pattern agrees well with that in the literature for OPC and AA concrete ,49,51. TOne unusual observation can be made from Meanwhile, most of the AA prisms, observed in cr), as shown in Equation (4). This normalized tensile stress is known as the tension-stiffening factor (β), where the post-cracking phase of this factor represents the tension-stiffening contribution [m is the average member strain, Pcr is the concrete cracking stress, and β is the tension-stiffening bond factor.Tension stiffening is commonly analyzed using the normalized tensile stress in concrete that was obtained using the average strain–stress approach suggested in the literature ,53,54,55ribution . This faribution ,49.(1)NThe diagrams reported in cr). This trend agrees well with that reported for LCFA prisms in the literature [An important observation from terature ,57. Anotterature . HoweverThe tension-stiffening factor (β) starts to decay at the end of the stabilized cracking stage, as shown in By referring to In contrast, the tension-stiffening factor of OPC  ratio on the tension stiffening was evaluated by employing the reinforcement of either 10 mm, 12 mm, or 16 mm in diameter. This corresponds to a Cc/db ratio of 3.25, 2.63, and 1.84, respectively. Referring to c/db ratio slightly improves the elongation resistance of the prisms by adding more stiffness to the bare bar response. This can be explained by the fact that increasing the confinement (Cc/db ratio) around the steel bar improves the tensile capacity of the surrounding concrete, which delays the formation of internal cracks [c/db ratio enhances the tension stiffening as increasing the confinement means increasing the effective concrete area that carries tensile stresses [In addition to the compressive strength, the effect of concrete cover to bar diameter  of the prism. Since many building codes formulas evaluate the cracking in terms of average crack spacing , the evolution of cracks in the current study followed a similar approach. Thus, the average crack spacing was plotted as a function of the average axial strain, as shown in A significant aspect of the investigation herein concerns crack spacing and its evolution to provide a clear understanding of concrete cracking. Thus, crack spacing results that include the maximum, average, and minimum crack spacing for all specimens were monitored during the test and reported in −6) and 1750 (10−6). However, Comparing the response of AA specimens with OPC counterparts in To better capture the influence of reinforcement ratio (ρ%) on the average crack spacing, the average crack spacing was plotted with respect to the reinforcement ratio (p%) in By referring to Many building codes included a 𝜙/p parameter in estimating the crack spacing of OPC concrete. Thus, it is worth mentioning that The crack patterns of all tested specimens are given in The crack width monitoring began upon crack initiation and continued whenever a new crack appeared on the concrete surface. After the crack pattern stabilized, the monitoring process was carried out at different loading intervals during the test with an increment that varied between 10 and 15 kN. An example of crack width monitoring is given in By referring to As given in The crack width growth results shown in This section thoroughly discussed the verification of the experimental crack spacing and width through comparisons with that computed using the OPC codes formula. Regressed models were also proposed, and their predictions were statically compared with OPC codes.One of the main goals of the current work is to check the applicability of OPC concrete approaches in various codes’ provisions by comparing the estimated values of the crack spacing with the experimental results of the present study. The design codes considered were EC2  and MC901 = 0.8). In contrast, the predicted values of the average and maximum crack spacing by CEB-FIP [By referring to  CEB-FIP  provisioc, bond factors, k1, and loading factor, k2, in the EC2 [The discrepancy in the predicted values between both design codes can be attributed to considering the concrete cover, C the EC2  formulat the EC2  formula max) based on the maximum crack spacing. In contrast, the ACI 224R [The expressions of EC2  and CEB-ACI 224R  approachAn overall comparison between the experimental and predicted maximum crack widths is summarized in max,exp) rather than using the theoretical maximum crack spacing . An improvement in code predictions has been obtained, and therefore, the crack width code formulas of EC2 [s of EC2  have beeComparing the experimental and the prediction values of the maximum crack width by the ACI 224R  provisioc/db ratios. Following this, the applicability of OPC concrete approaches in various codes of practice to predict the crack spacing and width of this concrete has been examined. Based on the results of the present work, these conclusions were drawn.Alkali-activated concrete is a sustainable construction material, and understanding its tension stiffening and crack spacing and width is of critical importance. These properties are essential indicators of the concrete suitability for structural applications, particularly at service limit states. Therefore, the current work aimed to investigate the tension stiffening and cracking performance of AA concrete by considering a wide range of compressive strengths and Ccr). Both concrete types also have a global response consisting of three stages: elastic, cracking, and stabilized. However, the OPC prisms experienced a brittle cracking mechanism, resulting in a sudden drop in the load–strain curves at the crack location.Both AA and OPC concrete prisms developed slightly similar axial cracking force  of AA concrete exhibited better ductile behavior than OPC concrete due to the strain compatibility between concrete and steel even after the crack ignition. In contrast, OPC concrete experienced a gradual reduction in the tension-stiffening factor (β) after the crack formation.c/db ratio) around the steel bar improves the tensile capacity of the surrounding concrete, which delays the formation of internal cracks and, consequently, enhances the tension stiffening of AA concrete.Compressive strength was generally found to be an influencing parameter in the global response of tested prisms because it improves the cracking resistance and bond between concrete and reinforcement. In addition, increasing the confinement  and the high bond behavior considered. In contrast, CEB-FIP predictions of crack spacing are mostly on the top side of the equality line. In addition, code provisions tend to underestimate the maximum crack width, especially EC2 , and thi


Temporomandibular disorders (TMD) are associated with musculoskeletal diseases (MSD), mental and behavioural disorders (MBD), and patients with TMD have been shown to have 2–3 times more days of sick leave (SL) and disability pension (DP) than the general population. MSD and MBD are two of the most common causes for SL and DP, and the association between TMD and the influence of comorbidities on the need for SL and DP among TMD patients need further clarification. This study investigates the impact of MSD and MBD comorbidity on SL and DP among TMD patients diagnosed in a hospital setting and/or surgically treated.All incident TMD patients diagnosed or treated in a hospital setting between 1998 and 2016 and aged 23–59 were included. A non-exposed comparison cohort was collected from the general population. The cohorts were grouped based on the presence of comorbidity: No comorbidity (Group I); MSD comorbidity (Group II); MBD comorbidity (Group III); and combined MSD and MBD comorbidity (Group IV). Main outcomes were mean annual days of SL and DP, and statistical analysis was conducted using generalized estimated equations.TMD subjects with no comorbidities (Group I) and with MSD/MBD comorbidity (Group II and III) were 2–3 times more often on SL and DP than the corresponding groups from the general population. However, in the group with both MSD and MBD comorbidity (Group IV), the difference between the TMD subjects and the general population was diminishing, suggesting an additive effect.TMD patients are more dependent on SL and DP benefits compared to general population and the difference remains even after considering MSD and MBD comorbidity. In individuals with combined MSD and MBD comorbidity, concurrent TMD has less impact on the need for social insurance benefits. The results accentuate the impact TMD has on the patients’ impaired ability to return to work and why TMD should be recognized as having a substantial impact on individual and economic suffering as well as on societal costs, with emphasis on the influence of comorbidities on patient suffering. Temporomandibular disorder (TMD) is associated with orofacial pain, dysfunction, and impaired quality of life and has an incidence rate of 4% in the general population , 2. TMD Globally, the leading causes of years lived with disability (YLD) are MBD, MSD, diabetes, chronic obstructive pulmonary disease and endocrine diseases . Low bacAn earlier study from SWEREG-TMD has shown that TMD has a great impact the patients’ need for both SL and DP, with mean annual days of SL and DP two to three times higher than the general population . The cloaThe National Patient Registry (NPR) was used to collect the data on incident TMD or surgical treatment from 1998–2016. The NPR was also used to collect MSD and MBD comorbidity from 1964 to the time of inclusion. The NPR is administered by the NBHW and includes 100% of all in-patient care and 80–100% of all out-patient visits, with information on both primary diagnosis and procedure codes. The positive predictive value of both in- and outpatient care is 85–95% .bThe Total Population Registry (TPR) was used to collect the unexposed comparison cohort . The TPRNational Board of Health and Welfare (NBHW)aThe Longitudinal Integrated Database for Health Insurance and Labour Market Studies (LISA) was used to collect information on SL, DP, and sociodemographic covariates. SL and DP is prospectively registered on an annual basis .Statistics Sweden (SCB)The details of this registry-based cohort study have been described earlier in previous publications from the SWEREG-TMD , 5, 12. Linking individuals between the registries was made possible through the use of the Personal Identification Number (PIN) . NBHW waMain exposure was collected from the NPR and defined as first registered TMD diagnosis (K07.6) and/or surgical treatment for TMD in a hospital setting between 1998 and 2016. First time of diagnosis or surgical treatment was considered as T0, meaning T0 could be any year during 1998–2016. Surgical treatment codes used as proxy for TMD were TMJ disc surgical procedure (EGB10), TMJ arthroscopy (EGA00), TMJ condylectomy (EGB00), TMJ prosthesis surgical procedure (EGC30), other surgical reconstruction of TMJ (EGC99), TMJ synovectomy (EGB20), TMJ biopsy (EGA20), injection of diagnostic or therapeutic substance in the TMJ (TEG10), TMJ condylotomy (EDC00), open reposition of TMJ luxation (EGC00), TMJ plastic surgery (EGC10), and TMJ plastic surgery with bone graft or other type of transplant (EGC20). The exposed cohort was divided into two sub-cohorts: non-surgical (NS)  and surgical (S) .The non-exposed (NE) comparison cohort was collected from the TPR and matched 1:10 for age, sex, and living area with the requirement of being alive at the time of inclusion. As working-age in Sweden typically spans from 18–65, only exposed and non-exposed subjects aged 23–59 at the time of inclusion were included in this study, to allow subjects to be monitored during the entire 10-year follow-up time.Comorbidities were previous diagnoses of MSD and/or MBD recorded in the NPR before the time of inclusion. ICD-10 codes were used to identify both MSD (M00–M99) and MBD (F00–F99) comorbidity. Data on comorbidity were collected from 1964 up to T0, i.e., all comorbidities were collected before first TMD-diagnosis or surgical treatment. ICD-10 was introduced in Sweden in 1997, and older versions of ICD-codes – i.e., ICD-7 (1958–1968), ICD-8 (1969–1986), and ICD-9 (1987–1996) – were translated to ICD-10 codes in accordance with instructions and a codebook provided by the NBHW.th chapter of ICD-10 (M00–M99), and MBD was defined as diagnoses within the 5th chapter of ICD-10 (F00-F99). The division into diagnostic blocks are described in Table MSD comorbidity was defined as diagnoses within the 13Group I: No comorbidity present,Group II: MSD comorbidity present, but no MBD comorbidity,Group III: MBD comorbidity present, but no MSD comorbidity, andGroup IV: Both MSD and MBD comorbidity present.To stratify on comorbidity, the following four groups of comorbidity presence were constructed:Outcome variables were SL and DP, collected from LISA between 1994 and 2017. SL is available for all Swedish citizens from the age of 16, and the first 14 days are reimbursed by the employer. DP is available for individuals aged 19–64 with lifelong incapacity of work. Both SL and DP are awarded in levels of 25, 50, 75 or 100% For both outcomes, net days were used – i.e., partial reimbursements recounted into full annual days. Only SL spells lasting more than 14 days were included since only those are included in the LISA registry. In their current forms, SL and DP are available in the LISA-registry from 1994.Covariates adjusted for in the model were: sex, level of education , country of birth , living area according to Eurostat’s Degree of Urbanisation (DEGURBA) , and age . Calendar year was also adjusted for, due to fluctuations in the policies of SL and DP insurance.The period of inclusion was 1998–2016, and the year of inclusion was defined as T0. Follow-up time for the outcomes SL and DP was five years before T0 (-T5) up to five years after T0 (T5). The subjects’ individual follow-up time of 10 years was included in the regression model. The follow-up time, the period of inclusion and the collection of comorbidities and outcome is depicted in Fig. All analyses were conducted using Stata v.16.1 software . Generalized estimating equations (GEE) was used to analyse number of net days on SL and DP during -T5 to T5 . SL and Figure The sociodemographic baseline data of the study sample as well as the exposure of comorbidities are presented in Table Table Figure Results from the GEE regression model adjusted for age, sex, educational level, country of birth, DEGURBA, and year of inclusion for SL are shown in Table For DP, the results from the GEE regression model adjusted for the same covariates are shown in Table In this study, we show that the higher dependence on SL and DP benefits among TMD patients compared to the general population remains even when considering MSD and MBD comorbidities. Irrespective of comorbidity, the TMD subjects had more mean annual days of SL and DP than the general population. There were some exceptions for certain MBD diagnoses such as F00–F09 , F20–F29 , and F70–F79 . That is, the non-exposed cohort had more annual days of DP than the exposed cohorts. The findings are important as they emphasize how work incapacity of TMD patients is strongly influenced by comorbidities and that the higher dependence on SL and DP benefits compared to the general population in Group I is most likely attributed to TMD. Therefore, the increased need for social insurance benefits among TMD patients cannot be solely explained by comorbidities.A previous SWEREG-TMD study showed that TMD patients are 2–3 times more reliant on SL and DP benefits than the general population, in particular patients with TMD that requires multiple surgical interventions . TMD relIn Sweden and the other Nordic countries, the healthcare system is mainly tax financed, so patients pay a very small fee to receive treatment. However, as the dental care system is tax financed to a considerably lesser extent, dental patients have higher out-of-pocket expenses . PatientIn a study focused on SL among patients with MSD, a large heterogeneity was seen between the different types of diagnoses, with the lowest number of SL days during a two-year period (2009–2010) for low back pain and myalgia and the highest in disc disorders and rheumatoid arthritis . The autIn this study, more comorbidities among TMD patients resulted in more days of SL and DP. This finding is not surprising and multiple comorbidities have been shown to affect the trajectory and need of social insurance benefits for other conditions as well –31. Our There is an intricate nexus of causal exposures that impact the need for SL and DP among any patient groups, including TMD patients. The impact of comorbidities, such as psychiatric-somatic diagnoses, on SL and DP and the ability to return to work have been established for many diseases –38. PsycThe most substantial strength of this study is the prospective nature of registry-collected data with no loss to follow-up, which the results are based on. The Swedish national registries contain high quality data that have high validity. As the number of subjects included in this study is unique, this may be the first study to investigate the impact of comorbidities on the use of social insurance benefits among TMD patients.The largest limitation of this study is that TMD represents a heterogenic patient group that in Sweden is primarily treated within the dental care system and therefore not available in the registries used in this study. Therefore, the non-surgical cohort represents a subgroup of all TMD patients who do not require surgical treatment for their condition, and the conclusions drawn about this subgroup may not be applicable to all TMD patients. Therefore, future studies should investigate how patients who are conservatively treated within the dental care system differ from the TMD patients diagnosed but not treated within the health care system. Furthermore, it is not possible to rule out that patients within the non-surgical cohort need surgical treatment later, causing a possible misclassification bias, specifically for subjects included late in the study. This study does not address whether earlier intervention, treatment, or prevention of severe TMD increases the patient’s ability to work, so studies on the matter are warranted. It is also important to address that the results might be less relevant in countries where private health insurances are used to a larger extent; nevertheless, the results indicate the burden of disease the TMD patients carry, specifically patients with several comorbidities.TMD patients were more dependent on SL and DP benefits compared to general population and the difference remained even after stratifying MSD and MBD comorbidity. In individuals with combined MSD and MBD comorbidity, concurrent TMD had less impact on the need for social insurance benefits. The results accentuate TMD patients’ impaired ability to return to work, specifically in the light of common comorbidities, and emphasize why TMD should be recognized as a disorder with a potential to substantially impact individual and economic suffering and societal costs.