Raspberry (Rubus spp.) is an economically important crop with a restricted growing season and very limited fruit shelf-life due to its extreme tenderness. In order to prolong its shelf life, an aqueous composition containing hexanal as the key active ingredient (HC) was applied as a preharvest spray during fruit development. The effects of HC were assessed using physiological, biochemical and anatomical parameters on the treated fruits and compared with the effects of mock inoculation which lacked hexanal. Sugars and acidity did not show a significant change in response to HC treatment, while the pulling force (the tension required to detach the berry from the receptacle) significantly improved in the HC-treated fruits, compared to control. Scanning electron microscope (SEM) analysis revealed a high correlation between the presence of rigid epidermal hairs and a stronger degree of attachment between berries and their receptacle in the HC treated fruits. Further, electron micrographs also showed abnormal crystalline depositions on the epidermal drupelets of the treated berries. Energy Dispersive X-ray Spectroscopy (EDS) analysis showed those crystals to be largely composed of calcium. HC treatment also resulted in the reduction of transcript level of three phospholipase D genes, as well as altered expression pattern of five members of the annexin gene family, and four calmodulin-binding transcription activators. Quantification of PLD activity showed that hexanal inhibited PLD activity in treated berries. The potential crosstalk between hexanal, phospholipase D activity and calcium and this crosstalk’s role in delaying fruit softening and in prolonging storage life of fruits shelf life is discussed. Rubus ideaus L.) is a tender fruit with an extremely short shelf-life. It is a rich source of health-promoting compounds such as anthocyanins. Rapid decline in fruit quality typically occurs due to decay and physiological breakdown, which is associated with the loss of membrane integrity leading to the progression of senescence and stress.1 Phospholipase D (PLD), a phospholipid-degrading enzyme is the key enzyme involved in initiating a cascade of catabolic events that leads to the eventual deterioration of the membrane, and is highly active in the fruits of other berry species such as strawberry.3Raspberry -β-D-glucanases, β-xylosidases, xyloglucanases, endotransglucosidases, endo-mannanases.7–11 However, efforts to suppress expression of cell wall-degrading enzymes by genetic engineering did not provide satisfactory results to efficiently prevent softening of fruits.13Suitable technologies and methods to enhance the postharvest shelf life of tender fruits are in high demand. The application of hexanal as a formulation, either pre- or postharvest, has shown promising results in enhancing the shelf-life of several fruits, vegetables and flowers.14 and tomato fruit,11 it was not as distinct in systems such as carnation flower petals.16 Thus, increased phospholipid degradation that occurs during ripening/senescence was linked to the activation of PLD by factors such as increase in cytosolic calcium and a decrease in pH, membrane binding promoted by C2 domain, and fatty acid retailoring that increases the availability of preferred PLD substrates.17 Altering the levels of exogenous calcium application affects parameters for senescence such as protein and chlorophyll content, respiration rates, and cell membrane fluidity.18 Other studies also reported that the addition of calcium rigidifies cell wall and obstructs enzymes such as polygalacturonase from reaching active sites.18 Calcium plays a special role in maintaining the cell wall structure in fruits and other storage organs by interacting with pectic acid in the cell walls to form calcium pectate. Thus, fruits treated with calcium are generally firmer. The irreplaceable nature of the calcium ion (Ca2+) as a signal transduction agent, and in cell wall polysaccharide interactions is undisputed; it is through these processes that calcium is central to stress responses, cell wall structure and remodeling, and to plant tissue development.20 It has been suggested that calcium rigidifies the cell wall by crosslinking pectic acid residues21 and stabilizing cell membranes.22 Nevertheless, accumulating evidence indicate that calcium is a universal second messenger, and plays an important role in plant growth and development by mediating response to a variety of environmental and hormonal signals.24 Signal triggered increase in cytosolic calcium are perceived by calcium sensors. Calmodulin is a ubiquitous calcium sensor, and present in all plant cell types examined thus far.26 It can modulate the actions of diverse target proteins involved in almost all aspects of cell activity including cell division, cell elongation, ion transport, secondary metabolism, plant defense, etc.26It has been suggested that inhibition of PLD activity by hexanal application could eventually lead to enhanced membrane stability, and thereby increased longevity of horticultural produce. Previous studies have attempted to correlate increased membrane deterioration that occurs during ripening and senescence to increased phospholipase D activity. Although such an increase in PLD activity was noticeable in some senescing systems such as broccoli florets27 Genome sequencing revealed that annexins in plants comprise a small multigene family with several members, eight in Arabidopsis thaliana28 and nine in Oryza sativa.29 The increased expression of certain annexins uunder specific developmental or environmental challenges has linked them with Ca2+ signalling during nodulation,30 ABA responses32 or cold acclimation.33 The possibility that annexins form Ca2+ channels, coupled with their presence in root hairs and the root elongation zone, suggests a role in generating the elevated cytoplasmic calcium concentration ([Ca2+]cyt) required for tip or extension growth. Plant annexins have been found to have in vitro ATPase and GTPase activity.34 AtANN1 has been identified in vitro as an ATP-binding protein.35Among the calcium-binding proteins, annexins form a family of structurally related proteins that exhibit calcium-dependent binding to phospholipids. Annexins are soluble proteins capable of Ca2+-dependent or independent association with membrane phospholipids.36 In this study, we report on understanding the mechanism of action of hexanal in delaying membrane deterioration and prolonging shelf life using multidisciplinary approaches including physiological, histological and biochemical characterization. The expression patterns of three phospholipase D genes, five annexin gene members, and four calmodulin-binding transcription activators, were quantified by qRT-PCR during fruit development and ripening in response to hexanal applications. Scanning electron microscopy as well as Energy Dispersive X-ray Spectrometry were conducted to analyze the structural and compositional details of treated and untreated fruits. In addition, phospholipase D activity was assayed during fruit development and six days postharvest after hexanal applications.Our previous study analyzed the effect of hexanal on the relative transcript abundance levels of a set of genes putatively associated with cell wall degrading enzymes in strawberry.Rubus idaeus (European red raspberry) grown under field conditions . For the molecular studies, fruit were selected at three developmental stages  and used as control. The study was performed for two consecutive growing seasons 2015 and 2016.2O was titrated with 0.1 M NaOH to pH 8.2 and expressed as percentage of citric acid (%). To evaluate the retention force needed to remove the torus from the fruit, we adapted a fruit hardness tester  with a 5 mm Penetrometer Tip. Fruit was attached to the instrument with a clamp and the force needed to remove its receptacle was recorded daily for twenty five fruits. Fruit weight was recorded daily for 25 fruits and the percent weight loss was calculated by taking the difference between initial weight and final weight divided by initial weight. After assessing physiological characteristics, all samples  were frozen in liquid N2 and stored at −80 °C for further analysis.To determine shelf life characteristics, fruits of similar size and developmental stage were harvested at commercial maturity. Several physical and chemical fruit quality attributes, such as pulling force (the required force required to detach the fruit from the receptacle), fruit weight, total soluble solids (TSS) and titratable acidity (TA) were evaluated daily through six days postharvest. TSS (%) of the raspberry juice was measured with a digital refractometer . For titratable acidity evaluation, 2 ml of fruit juice in 50 ml dH38 Quantitative reverse transcription PCR (qPCR) was conducted for 12 genes chosen to represent a variety of biological functions during the ripening and different developmental stages  for 30-minute durations. After the third change, samples remained in HMDS until all of the solution evaporated. Samples were sputter-coated with carbon using a Nanotek SEMprep 2 sputter coater. Imaging and quantification analyses were conducted using a Zeiss EVO LS15 EP-SEM. The Zeiss EVO SEM with LaB6 electron source has a resolution of ~100 nm and equipped with a Bruker energy dispersive X-ray spectroscopy (EDS) system with a silicon drift detector with a resolution of 123 eV and a 10 mm2 window area.Sample preparation was performed as described in Bray λex=530/λem=585 nm) product, proportional to the PLD activity in the sample. Three technical and three biological replicates were used in this assay, initial and final absorbance were measured at 570 nm using Epoch 2 Microplate . The slope of the calibrator curve was used to calculate the phospholipase D activity of the sample using the following equation:t=enzyme reaction time (20 min in standard assay) and n=dilution factor. Unit definition: one unit of PLD catalyzes the formation of 1 mmole choline per min under the assay conditions (pH 7.4).Phospholipase D Assay Kit was used following the manufacture’s recommended protocol . In this assay, PLD hydrolyzes phosphatidylcholine to choline, which is determined using choline oxidase resulting in a colorimetric (570 nm) /fluorometric. Significant differences were determined by Tukey’s HSD test with overall α = 0.05.Data from the two consecutive growing seasons (2015 and 2016) were used in this analysis. Statistical analyses were performed in SAS v9.4 . To meet assumptions of normality and heterogeneity of variance, data were logThe effect of hexanal on fruit quality, and physiological parameters that determine shelf life characteristics of raspberry fruits are provided in Fruit removal force, the pulling force required to remove a berry from its receptacle, was measured and used as an indicator of fruit softening. Hexanal application significantly increased the pulling force on the treated berry fruits compared to the control . All fruScanning electron microscope (SEM) analysis was conducted in order to better understand the mechanisms underlying the correlation between hexanal application and the enhancement of fruit retention. This analysis showed an abundant accumulation of epidermal hairs on the tip of the receptacles of the treated fruits while no epidermal hairs were observed on the tip of the untreated . EpidermThe expression patterns of three phospholipase D genes, five annexin genes and four calmodulin-binding transcription activator were quantified during fruit developmental and ripening in response to hexanal application. The five members of annexin group showed a constant increase in their transcript levels throughout the fruit development stages. . All annThe transcript levels of PLDs increased during fruit development, and attained their maximum transcript level at the mid-stage . The higA similar analysis of gene expression was conducted of these 12 genes throughout 6 days postharvest for hexanal treated and untreated fruits . The staIn order to strengthen our understanding of the effect of hexanal in altering fruit structure, photomicrographs, of the external (berry) and internal (receptacle) raspberry tissue samples were assessed during the different development stages using SEM. Hexanal applications at early or late stage of fruit development did not alter the external fruit structure . HoweverEnergy Dispersive X-ray Spectrometer (EDS) analysis was conducted in order to identify the nature of these crystal depositions. This analytical technique allowed us to conduct elemental analysis and to compare between two areas on the same sample. Na, Mg, Si, K and Ca were quantified in the external drupelets of the control samples and in the areas with or without the crystalline depositions present in the hexanal-treated samples. When these elements were quantified, Ca was the most abundant and distinguishable element in the crystalline structures appearing on the drupelets of the treated fruits . EDS anaIn conclusion, the effect of hexanal on altering fruit structure was development stage-dependent and exclusive on the exocarp and epidermal hairs.To evaluate the potential role of hexanal in the inhibition of phospholipase D and the extension of shelf life, PLD activity was assayed at various stages of development in raspberry fruits. In this colorimetric assay, phosphatidylcholine is converted to choline by PLD, where choline is then oxidized by choline oxidase. The intensity of the colored reaction product is directly proportional to the activity of PLD in the sample.One unit of PLD catalyzes the formation of 1 mmol choline per minute under the assay conditions (pH 7.4). PLD activity was analyzed by monitoring the liberation of choline by unit/l . The ear41 This is particularly true for raspberry, which is very tender by nature and has an extremely short shelf life. Biochemical and genetic studies of fruit softening indicated that cell-wall processing is the result of coordinated expression of several families of genes encoding cell-wall metabolism related proteins.7–11 However, many efforts to suppress expression of cell wall-degrading enzymes have not provided the insight needed to genetically engineer fruits whose softening can be adequately controlled.13 Phospholipase D (PLD), is the key enzyme involved in initiating a cascade of catabolic events that leads to the eventual deterioration of the membrane phospholipids.3 In this study, we tested the hypothesis that hexanal application could eventually suppress the PLD activity and thus lead to enhanced membrane stability, and thereby increased longevity of raspberry fruit. To achieve this end, we have used multidisciplinary approaches including gene expression, physiological and histological characterization and enzymatic activity in order to strengthen the understanding the effect of hexanal in enhancing raspberry fruit shelf life. Since the action of hexanal is so specific, there was little effect on the TSS and TA, while it had a striking effect on fruit retention. This is in in agreement with Anusuya et al.42 who indicated that pre-harvest spray of hexanal assisted in retention of mango fruits in the orchards for about 2–3 weeks. Extending the fruit retention time will significantly reduce the postharvest losses, resulting in a significant economic benefit. Moreover, this might be used as an indicator of enhanced fruit firmness especially since the growth regulators that target fruit retention may also be used to improve firmness, as both abscission and ripening are ethylene-mediated events in fruit development.43 In raspberry fruit there is some controversy as to whether the fruit are truly climacteric. Although there is a large increase in the rate of ethylene production, there is no accompanying burst in the rate of respiration.44 However, there are many features of fruit ripening that are common to both climacteric and non-climacteric species indicating that there is overlap in the molecular mechanisms underlying the ripening process in both types, despite the differences in respiration and ethylene production.The control of fruit softening is an important target for the improvement of fruit quality as fruit firmness appears to affect susceptibility to fungal pathogens.Part of the explanation of this increase in fruit retention was revealed by SEM analysis showing an abundant amount of the epidermal hairs on the tips of the treated fruit receptacles enhancing the attachment between fruits and their receptacles, thus increasing force required to remove the fruit receptacle.46 Calcium is recognized also as a critical element for rigidifying cell walls by cross-linking with pectins.47 However, to our knowledge, this is the first report suggesting a potential crosstalk between hexanal application and calcium accumulation. Gene expression data also supports this hypothesis as hexanal treatment increased the transcript levels of four annexins and three calmodulin-binding transcription activators during raspberry postharvest storage. Previous studies suggested the potential roles of annexins in fruit development and ripening. For instance, there was a positive correlation between the stage of fruit development in strawberry and transcript levels of two annexins genes.48 Data analysis from a proteome survey of strawberry fruits during ripening suggested a role of annexins in fruit ripening,49 however, an independent regulation of individual annexin was suggested probably due to the large protein family with multi-functions.Moreover, histological observations by SEM and EDS confirmed that there is calcium deposition on the epidermis of the drupelets of the hexanal treated fruit. Calcium has received considerable attention in recent years because of its desirable effects in controlling fruit ripening and quality by delaying ripening and maintaining firmness. For instance, increasing the calcium concentration of fruit through pre-harvest sprays and postharvest calcium treatments maintains firmness and prevents decay in both climacteric and non-climacteric fruits.51 One of the important findings in the study of Yang et al.51 was that the suppression of calmodulin genes could be critical for initiating ethylene-independent ripening process. In this study, the expression of three calmodulin-binding transcription activators  was differentially altered in response to hexanal application which reinforces the clues for a potential effect of hexanal on calcium binding proteins that function as signal sensors.Calcium-sensing proteins called calmodulins have been found to regulate both enlargement and ripening of tomatoes.1 These results conclusively showed that PLD expression was reduced during fruit ripening by the introduction of antisense PLD alpha cDNA in ‘Celebrity’ tomatoes. Hexanal treatment caused a clear PLD activity inhibition in raspberry fruits in a similar way that the antisense PLD tomato lines showed a reduced level of PLD activity. In the same report, microsomal PLD activity in strawberry fruit membranes was found to be stimulated by calcium. These results, together with our show a clear correlation between hexanal application, cellular calcium stimulation and PLD activity inhibition. However, the complete sequence of signal transduction events involved in this link requires further investigation.Our studies also demonstrate a clear correlation between PLD activity, and PLDs gene expression at various stages of raspberry fruit development as well as during the storage period. A similar analysis was conducted using normal and antisense PLD in ‘Celebrity’ tomatoes.et al.39 also indicated that the stage of fruit application is crucial since PLD inhibition has very little effect once the membrane starts to deteriorate and lipid catabolic cascades are initiated. For example, tomato fruits are treated at the mature green stage to obtain the best results in terms of colour and flavour development. Whereas berry fruits such as cherry should be treated about 15 days before harvest to maximize their shelf life to 30 days at 4 ° C.5Despite the fact that raspberries appear to be a very responsive system to hexanal application, the stage of application of pre-harvest hexanal treatment seems to be very important in order to modulate the PLD inhibition and to achieve optimum effect. For instance, the histological observations reveal that the calcium depositions will occur exclusively if the fruits are treated in their mid-development stage. Misran 52 These two characteristics should alleviate any fears consumers might have on its use on edible produce such as raspberry. It would be also interesting to determine if the calcium accumulation due to hexanal application will affect the edible quality and taste of fruits. Further studies could be used to develop an understanding of the biochemical mechanism of interaction between hexanal and the intercellular calcium in the overall regulation of PLD in extending fruit shelf life.In conclusion, this work clearly demonstrated the important role hexanal can play in reducing PLD transcript levels and activity as well as altering some of the calcium binding proteins that function as signal sensors. Thus, hexanal application might be a desirable and suitable technology to enhance the postharvest shelf life of raspberry in light of its extremely perishable nature. Hexanal, in spite of its chemically inclined name, is a natural compound produced by all plants as a volatile defense mechanism. It is also a GRAS  compound approved by the FDA as a food adjunct.


In this paper, the relationship between the static and dynamic elastic modulus of concrete and the relationship between the static elastic modulus and compressive strength of concrete have been formulated. These relationships are based on investigations of different types of concrete and take into account the type and amount of aggregate and binder used. The dynamic elastic modulus of concrete was tested using impulse excitation of vibration and the modal analysis method. This method could be used as a non-destructive way of estimating the compressive strength of concrete. The compressive strength and modulus of elasticity are the most important properties of concrete from the viewpoint of structural design. Commonly, these parameters are determined by the uniaxial compression of cylindrical or cube-shaped specimens, according to standard procedures, and so values obtained that way are considered to be the reference values. However, it is not always possible to use these methods in practice because they are destructive and require the collection of numerous test samples during concreting operations. In addition, properly conducted modulus of elasticity testing using cylindrical specimens is a relatively time-consuming process. Therefore, we seek non-destructive methods in order to estimate these parameters for hardened concrete, such as: the sclerometric method, the ultra-sound pulse velocity method, and the impulse excitation and modal analysis method.Technological developments and better access to more appropriate apparatus has resulted in significant progress in the dynamic testing methods used in structural health monitoring. These methods could also be used in concrete material parameter testing  or to monitor its increase over time. A significant disadvantage of non-destructive methods is the fact that the values are achieved indirectly, i.e., the results of each test have to be converted to a certain parameter  using a previously assumed relationship between these values. The natural frequencies obtained using impulse excitation testing and modal analysis methods are very good indicators of the state of concrete elements because the fundamental resonant frequency of vibration decreases with an increasing degree of material degradation . This alThe most commonly used equations were presented by Neville (2000) . SelecteThe equations mentioned above are convenient to use when calculating the compressive strength for a known modulus of elasticity because the compressive strength and concrete density are the only parameters required however, their accuracy in predicting the modulus of elasticity value is not sufficient for all kinds of concrete. This indicates that the relationship between elastic modulus and compressive strength is influenced by a number of factors, such as: the type of aggregate, humidity, age of the concrete, and the type of binder used. As preliminary research shows , the equThe effect of aggregate type on the elastic properties of concrete is widely discussed in the literature e.g., ,13. ThesA comprehensive analysis, based on the results of more than 3000 concrete samples, is presented in paper . Those aAn interesting model is presented in . The M5′The values calculated using the equations proposed in the literature usually provide satisfactory results for specific kinds of concrete only. On the rare occasion that an attempt has been made to provide relationships that take into account the influence of binder and aggregate type, the results have been significantly different from the ones determined experimentally. Additionally, most of the equations proposed in the literature refer to ‘normal’ strength concrete, although high strength concrete is also investigated .An additional problem when calculating the compressive strength of concrete using the impulse excitation method is the difference between the static and dynamic modulus of elasticity of concrete. The studies clearly show that the dynamic modulus is significantly greater than the static one. It could be the result of large differences in the strain rates associated with the static and dynamic testing methods  or it coThe literature survey revealed equations that describe the relationship between the static and dynamic modulus of elasticity. The most common are equations proposed by Lyndon and Balendran , PopovicThe relationship between static and dynamic modulus was investigated e.g., in ,24,25. TThe comprehensive study of the relationship between dynamic elastic modulus and compressive strength of concrete is presented in . The autThe method of dynamic elastic modulus testing preferred in this paper is based on excitation of a concrete specimen using a hammer and then analysis of its dynamic response. Recommendations for concrete testing methods using impulse excitation methods and modal analysis can be found in  and the It is most convenient to test beam-shaped samples of concrete. Commonly, these specimens are supported in vibration nodes to form a so-called ‘free-free’ configuration . In the Non-destructive methods of concrete testing are commonly based on experimentally derived relationships. The accuracy of the methods mentioned is highly dependent on the assumptions made. The literature survey, as well as experimental tests, demonstrated that widely used relationships are not appropriate to all types of concrete, especially in the case of concrete containing mineral additives.In case of concrete at early-age, values of compressive strength and elastic modulus increase significantly. In practice, it is of interest to know when the framework can be removed or when a construction element can be loaded without excessive deflections or cracking. Therefore, non-destructive methods are highly demanded in the situ testing. The accuracy of the estimated values in this way depends on the relationship between dynamic elastic modulus (obtained from the non-destructive testing), static elastic modulus, and compressive strength. In this paper, the investigation of different concrete compositions and properties, including different types of cement matrix, is presented. Based on static and dynamic elastic modulus and compressive strength testing results, the relationship between these parameters was formulated, taking into account the age of the concrete and the type and volume content of each ingredient . The impulse excitation and modal analysis method were used as a non-destructive method to determine the dynamic elastic modulus of concrete. It should be noted that values of initial and stabilized elastic modulus (introduced recently in EN 12390-13:2014 standard) were included to the proposed relationship between elastic modulus and compressive strength of concrete, which is the novelty in this field.In order to formulate the relationship between dynamic elastic modulus and compressive strength, different types of concrete were investigated. The tested concrete compositions varied in terms of the amount and type of each aggregate and also the type of binder. The chemical composition of the cements and metakaolin used is presented in In concrete mixes M1–M3, the same aggregate type  and the same volume content of aggregate were maintained. These mixes varied in the type of binder used. In order to increase the effect of cement type on the elastic properties of the concrete, an increased binder content was applied. This was carried out on self-compacting concrete (M4 and M5) and high strength concrete (M6).The concrete mixes M4 and M5 varied only by the aggregate type. The same volume content of basalt aggregate (M4) and natural aggregate (M5) was applied. The composition of concrete M1–M6 is presented in The dynamic elastic modulus was determined using procedures based on EN ISO 12680-1:2008. In this method, the vibration of the concrete beam is induced by a random force hitting the beam. The recorded signal is subjected to fast Fourier transform (FFT) and the fundamental resonant frequency was then determined.The dimensions of the concrete samples were: 500 mm length, 100 mm width, and 100 mm height. Each specimen was supported by steel tubes, positioned in such a way so as to coincide with the theoretical occurrence of fundamental vibration mode nodes. An illustration of the apparatus used in the dynamic elastic modulus testing is presented in 4, and The values of dynamic elastic modulus were calculated using the analytical relationship between the rigidity of the test sample, its geometry and mass and the fundamental resonant frequency, using the equationrding to .Compressive strength was determined using the method described in European standard EN 12390-3.CE,0 and CES,, respectively). The parallelism of the top and bottom surfaces was obtained using the steel caps and fine sand layer method, in accordance with Appendix ‘A’ of PN-EN 12390-3.The static elastic modulus was determined using cylindrical specimens—150 mm diameter and 300 mm high—in accordance to EN 12390-13:2014. Standard method ‘A’ was applied and then two kinds of static elastic modulus were determined: initial and stabilized  in days and the coefficient depends on the binder type . The largest differences between the calculated and experimental values are approximately 30%. This indicates that the relationship between static elastic modulus and compressive strength is influenced by the type of binder used. Thus, the parameter related to binder type and volume content was introduced to Equation (7)Taking into account the coefficients that are dependent on the type of aggregate and binder , the comThe values of the  results  and they2 = 0.950. The comparison of calculated compressive strength (according to ASTM 318-95 and using the relationship proposed in this paper) with the experimentally obtained values is presented in Based on the investigations carried out, it can be stated that the proposed relationship allows the compressive strength of concrete to be calculated with better accuracy. The differences between calculated values and those obtained experimentally a indicatBased on the results of our investigation, the relationship between the static and dynamic elastic modulus and the relationship between the static elastic modulus and compressive strength of concrete were formulated.The proposed relationship between dynamic elastic modulus and compressive strength takes into account the type and amount of aggregate and binder type. It was demonstrated that consideration of these factors allows compressive strength to be calculated with better accuracy.The investigation results showed that the stabilized elastic modulus is proportional to the initial elastic modulus and that the proportionality coefficient between them is dependent on the type of aggregate used.The proposed relationship between dynamic elastic modulus and compressive strength could allow the use of impulse excitation and modal analysis method as a non-destructive method of compressive strength testing for concrete. However, the relationship should be developed through further experimental analysis of the results achieved for different types of concrete.


Since the construction sector uses 50% of the Earth’s raw materials and produces 50% of its waste, the development of more durable and sustainable building materials is crucial. Today, Construction and Demolition Waste (CDW) is mainly used in low level applications, namely as unbound material for foundations, e.g., in road construction. Mineral demolition waste can be recycled as crushed aggregates for concrete, but these reduce the compressive strength and affect the workability due to higher values of water absorption. To advance the use of concrete rubble, Completely Recyclable Concrete (CRC) is designed for reincarnation within the cement production, following the Cradle-to-Cradle (C2C) principle. By the design, CRC becomes a resource for cement production because the chemical composition of CRC will be similar to that of cement raw materials. If CRC is used on a regular basis, a closed concrete-cement-concrete material cycle will arise, which is completely different from the current life cycle of traditional concrete. Within the research towards this CRC it is important to quantify the benefit for the environment and Life Cycle Assessment (LCA) needs to be performed, of which the results are presented in a this paper. It was observed that CRC could significantly reduce the global warming potential of concrete. For CRC2, T(0.50) and T(0.45) service times of at least 100 years were obtained for all calculations. Regarding chloride induced corrosion, the service life of 100 years was found valid for all four mixtures.Nonetheless durability tests were performed to verify the potential service life for concrete exposed to carbonation or chlorides, whereby rebar corrosion can be initiated. The results of this durability assessment can be found in De Schepper . Based oWith the obtained service times the functional units were calculated. Within the calculations the service time was given a maximum value of 100 years since this was the designed minimum life span. The results are presented in Most of the data used in this study were taken from the ecoinvent 2.0 database , which w2 emissions related to the chemical decarbonation of limestone.Within the ecoinvent 2.0 database , the cemThe process “Concrete, normal, at plant/CH U” from the ecoinvent 2.0 database  was adapThe life cycle inventory considered for the fly ash is based on . Fly ashThe replacement of 20%–30% of natural concrete aggregates by recycled concrete aggregates will have no significant impact on the durability performance of concrete ,19. For 3 concrete was considered. The energy needed for placing and compaction of the concrete was neglected according to De Herde and Evrard [et al. [The concrete produced at the concrete mixing plant is subsequently transported to the construction site. According to De Herde and Evrard , the aved Evrard . It shou [et al.  estimateThe environmental impact of concrete within the use phase ) is highly depending on the type and application of the built concrete structure or component. These issues are of particular interest in case different construction materials are studied, in example for a specific case study. Since this paper focuses on the material of concrete itself, the environmental impact of the use phase was not considered since a comparable impact is expected for the different concrete types.The data for demolition of the construction and sorting of the waste was taken from the ecoinvent 2.0 database , namely For CRC, all concrete waste is recycled within the cement production process. Since the geographical spread of concrete mixing plants and sorting plants was found comparable, the average distance between the construction site and a sorting plant was also assumed to be 50 km. The average minimum distance from a Flemish sorting plant to a cement plant was calculated to be 90 km.2 emission is expected within the regeneration process of cement from CRC instead of a traditional cement raw meal. Thermogravimetric analysis of CRC raw meal showed a total mass loss of about 35 wt%. The weight loss of 30 wt% between 600 and 900 °C was considered to be CO2. The raw material CO2 emissions from CRC are, thus, expected to be 0.46 g/g clinker, which is indeed about 15% lower than 0.54 g/g clinker in case of the traditional clinker production process. In practice, part of the CRC will be carbonated, and higher CO2 emissions can be expected. However, since this CO2 was captured from the atmosphere during the life cycle of concrete, this additional CO2 release was considered neutral in the life cycle assessment. Finally about 1.54 kg CRC will be needed for the production of 1 kg clinker, since the total mass loss measured by thermogravimetric analysis was about 35 wt%.Due to the presence of non-carbonate CaO, mainly from the cement, a lower raw material COThe waste scenario of traditional concrete was analysed according to two alternative routes, namely recycling as concrete aggregate or final disposal, e.g., as (sub)base material, for embankment constructions and in earth construction works. For the final disposal of concrete waste (including transport), the data were taken from the ecoinvent 2.0 database , namely 2 from the atmosphere, which can be seen as a benefit when performing a Life Cycle Assessment. In case of a built concrete structure, the ability for capturing CO2 is negligible, however in case of demolished concrete, the impact is more significant. Collins [2 emissions can be overestimated by 13%–48% depending on the type of concrete and application of the recycled concrete aggregate (RCA) during the secondary life. Since this impact is not easily quantified and is highly depending the type of concrete and application of the RCA, this CO2 capture from the atmosphere was not considered within the traditional recycling scenario.As already mentioned, concrete is able to capture CO Collins  estimateFor the impact assessment the LCA software SimaPro was used. This software is developed by PRé Consultants (Product Ecology Consultants) in the Netherlands and it contains the complete ecoinvent 2.0 database . The assThe results of the life cycle impact assessment of the cement production process for the different impact categories are presented in Taking a first look at the results, it is seen how the environmental impact of the cement production process can be reduced by incorporating supplementary cementitious materials, such as Blast Furnace Slag. Depending on the considered impact category, the environmental impact of the cement production can be reduced with 22%–47% by making use of BFSC instead of OPC. When replacing part of the clinker by BFS, only some heat (from fuels) and additional energy for grinding (in operation of the cement plant) is required for its pre-treatment. The environmental impact of this treatment is of course significantly lower compared to the whole clinkering process, explaining the reduced environmental impact of BFSC.2 emissions, related primarily to the clinkering process. Looking at 2 emissions for clinker production are related to the decarbonation process of the raw materials ”). Additionally 36% is related to the burning of fuels; 33% from alternative fuels of which the CO2 contribution is added to the clinkering process ” in The cement production process is known for its high COFor the other impact categories, it is seen that the environmental impact is dominated by the use of fuels and the air emissions related to the clinkering process. Furthermore the required electricity  and the consumables  necessary for the operation of the cement plant contribute significantly to the human toxicity and aquatic ecotoxicity, and, to a smaller extent, also to the terrestrial ecotoxicity and eutrophication of the environment.3 at a construction site are shown. The Functional Unit is thus not considered yet, but the transport from the concrete plant to the construction site is incorporated into the calculations.In It is seen that depending on the considered impact category, both CRC and traditional concrete mixtures can have the lowest environmental impact. The main advantage of using CRC2 is related to the global warming potential, which is 30%–34% lower compared to the traditional concrete mixtures. The good performance of CRC2 regarding the global warming potential is due to its reduced clinker content by using both fly ash and Blast Furnace Slag Cement. Considering global warming, the environmental impact of CRC1 and the traditional concrete are comparable.Regarding the Ozone layer depletion, CRC2 has a slightly better performance compared to the reference concrete mixtures, with a reduction of 6%–10%. Again, the result for CRC1 is comparable with the traditional concrete mixtures. While the global warming potential is strongly related to the cement content of the concrete, the ozone layer depletion is not only related to the cement content, but also to the required transport of the materials.The environmental impact of the concrete mixtures regarding the other impact categories is mainly related to their cement and fly ash content and the required transportation. To a smaller extent also the sand and aggregates, the superplasticizer, if needed, and the operation of the concrete mixing plant contribute to these impact categories. In case of abiotic depletion, acidification, photo-oxidant formation and the terrestrial ecotoxicity, the environmental impact of CRC2, T(0.45) and T(0.50) are comparable, while the impact of CRC1 is 8%–26% higher. The environmental impact considering eutrophication, human toxicity and aquatic ecotoxicity of both CRC mixtures is worse compared to the traditional concrete mixtures. The impacts are 25%–47%, 18%–25%, and 53%–72% higher, respectively.The results of the impact assessment of the end-of-life scenarios for both CRC and traditional concrete mixtures are presented in 2,eq emissions are indeed several times higher when CRC is recycled in the clinkering process, the release of a significant amount of CO2 is also avoided. Indeed, for each 1 kg CO2,eq released upon the recycling of CRC in a clinker kiln, the emission of 1.08 CO2,eq related to the burning of a traditional cement raw meal is avoided. The latter will reduce the impact on global warming of the CRC concrete mixtures significantly.The results regarding global warming definitely stand out for the CRC recycling opportunity. Although the CONonetheless, for the other impact categories, the recycling of the concrete waste seems not always better compared to its final disposal. The main contributor to the environmental impact of the recycling of both CRC and traditional concrete is the transport of the materials. In total, 140 and 50 km of road transport was considered for the recycling of CRC and traditional concrete, respectively, while the ecoinvent 2.0 data used for the final disposal of concrete takes into account a transport distance of only 15 km. Indeed, concrete rubble is often disposed in the neighbourhood as (sub)base material, for embankment constructions and in earth construction works. Depending on the considered impact category, the transport contributes for 57%–88% and 24%–66% to the environmental impact of the recycling of CRC and traditional concrete respectively, while it only amounts to 10%–25% in case of the mere final disposal of the concrete waste. It will, thus, be beneficial for both recycling of CRC and traditional concrete to reduce the environmental impact of the transport by minimizing road transport and maximizing rail and barge transport. As can be expected, the main impact of the final disposal of concrete is related to its disposal to an inert material landfill (47%–69%).The impact of the demolition process is identical for all end-of-life scenarios and amounts to 8%–27%, 5%–32%, and 16%–41% in case of CRC recycling, recycling of traditional concrete as aggregates, or final disposal of concrete waste, respectively. The additional processing needed to sort the waste materials for both recycling options is rather limited, except for the impact categories eutrophication, human toxicity, aquatic ecotoxicity and terrestrial ecotoxicity, wherein contributions of 5%–38% and 9%–45% were calculated for CRC and traditional concrete recycling, respectively. The additional processing that should be considered for the aggregate production in case of the recycling of traditional concrete varies between 8% and 41%, depending on the considered impact category.In the previous part of this chapter it was seen that most impact categories of the concrete production process are dominated by the cement manufacturing process and the required transportation. In case of abiotic depletion, acidification, eutrophication, photo-oxidant formation, human toxicity and aquatic toxicity, the environmental impact of fly ash might not be underestimated. The environmental impact of the sand and aggregates used in concrete is limited.2 emissions. The main goal for the development of a sustainable concrete is thus minimizing its clinker content , while still obtaining a high performance .The underlying cause of the huge environmental impact of the cement production is the clinkering manufacturing. Indeed, high amounts of raw materials and fuels are needed for this process, which goes along with high COAdditionally, the recycling possibilities of concrete will affect its sustainability. It was seen that additional transport and processing costs are needed for the recycling of concrete, nonetheless, natural resources will be saved and waste disposal is avoided. Comparing the recycling of CRC and traditional concrete, it was observed that although the global warming potential is lower, mainly the longer transport distances in case of CRC increase its environmental impact.The question, thus, remains whether the whole CRC concept is indeed sustainable, when looking at its whole life cycle. The life cycle impact assessments of the different stages in the life cycle of concrete described in Looking at the results for traditional concrete, it is seen that the environmental impact in case of both recycling as aggregate or the disposal of the concrete is comparable regarding most impact categories . The impact of concrete disposal is higher when looking at the ozone layer depletion, while the impact on the ecotoxicity is higher for the recycling as concrete aggregate.2 emission of CRC raw material compared to traditional cement raw meals due to the presence of CO2 free lime in a CRC cement raw meal. The clinker content and strength performance of CRC1 are, however, not low or high enough, respectively, to obtain a lower environmental impact compared to the traditional concretes. Depending on the obtained service life, the environmental impact will be 5%–88% (100 years), 31%–134% (81 years), or 68%–200% (63 years) higher compared to the disposal of concrete and 1%–64% (100 years), 26%–104% (81 years), or 61%–161% (63 years) in comparison with recycling concrete as aggregate. Only regarding the global warming potential, the environmental impact might be 7%–35% lower if a service life between 81 and 100 years can be obtained. With a service life of 63 years, the global warming potential is 4%–20% higher compared to T(0.50) and T(0.45).Both the low clinker content and the good strength performance of CRC2 compensate for the additional environmental costs related to its recycling as cement raw meal for most impact categories . While in most cases, the impact of CRC2 is comparable to the traditional concrete mixtures, the benefits regarding the global warming potential are significant and a reduction of 66%–70% was calculated. The lower global warming potential for the CRC mixtures is the result of the lower COCompletely Recyclable Concrete (CRC) is designed to be recycled within cement production, reducing the environmental impact of concrete and cement production. In order to prove the benefits for the environment of this CRC a life cycle assessment was conducted.2 free CaO. Thus making use of CRC instead of a traditional cement raw meal for the manufacturing of clinker will avoid the emission of a certain amount of CO2.The environmental impact of the concrete production process was found to be strongly related to its binder content  and the required transport. Also the recycling opportunities of concrete are strongly affected by the required transport, which is in case of CRC expected to be higher, as transport to the cement plant is expected to be longer compared to the distance to a concrete recycling plant. The main environmental benefit of CRC recycling is related to its global warming potential. Compared to a traditional cement raw meal, CRC will contain a certain amount of COLooking at the complete life cycle of CRC and traditional concrete it was found that regarding the global warming potential a reduction of 66%–70% is possible when a high strength CRC with a low clinker content is designed. In case of a normal strength CRC with a higher clinker content, the reductions can be up to 7%–35% if a sufficient service life can be obtained. For most other impact categories  only the performance of a high strength CRC with low clinker content could compensate for the additional transport required in the recycling process.


Objectives To examine the impact of use of rapid diagnostic tests for malaria on prescribing of antimicrobials, specifically antibiotics, for acute febrile illness in Africa and Asia.Design Analysisof nine preselected linked and codesigned observational and randomised studies .Setting Public and private healthcare settings, 2007-13, in Afghanistan, Cameroon, Ghana, Nigeria, Tanzania, and Uganda.Participants 522 480 children and adults with acute febrile illness.Interventions Rapid diagnostic tests for malaria.Main outcome measures Proportions of patients for whom an antibiotic was prescribed in trial groups who had undergone rapid diagnostic testing compared with controls and in patients with negative test results compared with patients with positive results. A secondary aim compared classes of antibiotics prescribed in different settings.Results Antibiotics were prescribed to 127 052/238 797 (53%) patients in control groups and 167 714/283 683 (59%) patients in intervention groups. Antibiotics were prescribed to 40% (35 505/89 719) of patients with a positive test result for malaria and to 69% (39 400/57 080) of those with a negative result. All but one study showed a trend toward more antibiotic prescribing in groups who underwent rapid diagnostic tests. Random effects meta-analysis of the trials showed that the overall risk of antibiotic prescription was 21% higher  in intervention settings. In most intervention settings, patients with negative test results received more antibiotic prescriptions than patients with positive results for all the most commonly used classes: penicillins, trimethoprim-sulfamethoxazole (one exception), tetracyclines, and metronidazole.Conclusions Introduction of rapid diagnostic tests for malaria to reduce unnecessary use of antimalarials—a beneficial public health outcome—could drive up untargeted use of antibiotics. That 69% of patients were prescribed antibiotics when test results were negative probably represents overprescription.This included antibiotics from several classes, including those like metronidazole that are seldom appropriate for febrile illness, across varied clinical, health system, and epidemiological settings. It is often assumed that better disease specific diagnostics will reduce antimicrobial overuse, but they might simply shift it from one antimicrobial class to another. Current global implementation of malaria testing might increase untargeted antibiotic use and must be examined. There is wide recognition that overuse of antimicrobials drives resistance in micro-organisms.8Across tropical and subtropical zones in Africa and Asia, acute febrile illness is one of the most common reasons for people to seek treatment from health services.15We hypothesised that improved malaria diagnosis to reduce use of antimalarials, a widely used antimicrobial class, could paradoxically drive an increase in untargeted use of other antimicrobials, such as antibiotics, particularly when test results for malaria are negative.35www.actconsortium.org) included several studies that evaluated the potential of rapid diagnostic tests for malaria to improve case management for patients with undifferentiated fever in malaria endemic areas. Data from these studies, conducted in multiple geographical, epidemiological, and health system settings, provide the largest and most varied sample to date to assess whether changes in antimalarial prescribing behaviour are associated with shifts in antibiotic prescribing. To inform policy for treatment of febrile illness, we compared settings where tests were and were not implemented, examined the differences in antibiotic prescribing overall and by test result, and identified the antibiotic classes used in different settings.The ACT Consortium  and with different approaches to implementation. To avoid selection bias, our combined analysis includes all studies in the ACT Consortium that were designed a priori to test the effect of introduction of rapid diagnostic tests for malaria on prescribing of antimalarial drugs, where providers could prescribe antibiotics, and did not include any other studies post hoc. Detailed descriptions of the individual studies are available in open access publications.43We included in the analysis studies that met the following criteria: evaluated an intervention to implement rapid diagnostic tests for malaria in settings where participating providers could prescribe both antimalarials and antibiotics, compared sites with and without the intervention, documented prescriber behaviour as a primary outcome, and collected individual patient data on diagnostic test results and treatments prescribed including antibiotics. Tables 1 and 2The studies were conducted in 2007-13 in Afghanistan, Cameroon, Ghana, Nigeria, Tanzania, and Uganda in a mix of rural and urban settings. Rapid diagnostic tests were introduced among government sponsored community health workers ), in public health facilities only Afgh-pub,, in privMicroscopy services were not present or were limited in five study settings.Prescribing data were collected through patient exit interviews or records of treatments administered (“registers”) completed by the provider, both of these methods, and both registers and follow-up interviews for a subset of patients (Uga-priv).41The main outcome of interest was the proportion of patients in each setting who were prescribed at least one systemic  antibiotic. Other outcomes included the type of antibiotic prescribed.The development of the primary research studies, from which these data are drawn, was informed by formative research among health workers, community members, and other stakeholders in the study settings; details for individual studies are available in open access publications.We performed three analyses to represent different policy and clinical perspectives. The main outcome for the first two analyses is the risk ratio of being prescribed at least one systemic  antibiotic. The first analysis compared overall antibiotic prescribing for each study, in settings with and without rapid diagnostic test interventions; this represents the overall policy effect of introducing diagnostic tests in a given context. The second, restricted to patients in intervention settings, looked at those with positive test result compared with those with negative results. This analysis shows the effect of test result on antibiotic prescribing. In the third analysis, we categorised prescribed antibiotics by drug class to explore the range of antibiotic classes used in different settings. We also examined the impact of rapid diagnostic tests on prescription of the most commonly used antibiotic classes, defined as those that were prescribed to at least 5% of patients in at least one site, to see if there was a differential effect of test result by antibiotic class.To allow comparison of the impact of introduction of rapid diagnostic tests for malaria and test results across studies, we calculated the risk ratios with their 95% confidence intervals for each study using binomial regression with a log link. The Huber-White robust standard error was used to account for correlation within the highest level of clustering .Table 1Of patients in intervention settings for whom a malaria test result (slide or rapid diagnostic test) was available, 61 324/157 345 (39.0%) tested negative for malaria, ranging from 30.5% (35 711/117 070 in southeast Uganda) to 99.8% (462/463 in northern Afghanistan). Parasite prevalence acts as a proxy for local endemicity.Table 32=65%).Antibiotics were prescribed to 127 052/238 797 (53%) patients in control groups and 167 714/283 683 (59%) in intervention groups. The proportion of patients prescribed at least one systemic  antibiotic ranged from 18.2% (110/605) to 72.8% (287/394) in control settings and from 15.2% (752/4946) to 78.5% (1130/1439) in settings with a rapid test intervention. Relative to control settings, the proportion of patients receiving an antibiotic prescription was higher or had a trend towards being higher where rapid diagnostic tests were introduced in all but one of the studies (in Nigeria), with risk ratios ranging from 0.65 to 2.98  was higher for patients with negative test results across most sites  in any of the study sites that might have affected the results.It is not possible to know whether antibiotic prescription was appropriate at the individual patient level in this analysis because the studies did not collect full clinical data or samples for further laboratory investigation. In most similar settings where bacterial diagnosis has been undertaken, however, few patients have documented bacterial infections; fewer than 2%  of patients with uncomplicated febrile illness have positive results on blood cultures.Case management guidelines for limited resource settings, such as the WHO’s Integrated Management of Childhood Illness (IMCI) and Integrated Management of Adult and Adolescent Illness, do not recommend empirical use of antibiotics for non-severe febrile illness of unclear aetiology.57Haemophilus influenzae type B, and the resulting reduction in bacteraemia and bacterial infection, have further reduced the risk that non-specific, non-severe febrile illness is caused by a potentially serious bacterial pathogen in many African and Asian countries.60With declining incidence of malaria in many settings, the proportion of fevers attributable to illnesses other than malaria stands to increase. Currently rapid diagnostic tests are more useful to rule out malaria than to rule it in, at least in Africa, but this is likely to change as the incidence of malaria drops. As this transition occurs and the proportion of negative test results increases, the risk of inappropriate antibiotic treatment of patients with negative results is likely to increase as well. Widespread childhood vaccination for pneumococcus, meningococcus, and Even when treatment with antibiotics is warranted, patients might not receive the most appropriate antibiotic for their illness, particularly in settings with inadequate healthcare infrastructure.Streptococcus pneumoniae isolates are resistance to penicillin, while 0% to 35% isolates of non-typhoidal salmonella are resistant to fluoroquinolones.65Within each study site, different antibiotics predominated, which could reflect to varying degrees local availability including stockouts  of antibiotics because of intermittent supply,Clinical case management typically follows an expected pathway that culminates in the prescription or purchase of medicines.41This analysis suggests that while introduction of rapid diagnostic tests for malaria can reduce untargeted excessive use of antimalarials—a highly beneficial public health outcome across malaria endemic regions—it can also have the unintended consequence of driving up untargeted and probably excessive use of antibiotics. In this analysis, the shift included antibiotics from several classes and was consistent across nearly all the varied clinical and epidemiological settings studied, increasing the generalisability of the findings. Therefore when rapid diagnostic tests are introduced, policymakers and clinicians should avoid a switch to overuse of antibiotics, a concern that increases the challenges of changing prescribing practice. This awareness is important in the design of programmes for provider training and community education in Africa and Asia, where antibiotic use is already relatively unregulated and unrestricted.This analysis shows quantitatively that introduction of rapid diagnostic tests for malaria can lead to an increase in antibiotic prescribing in many settings. Two major gaps in evidence need to be filled to inform policy and clinical practice guidelines to deal with this problem. The first is to identify the current drivers of this prescribing behaviour  to inform efforts toward behaviour change. The second, especially important for the rational revision of diagnostic algorithms such as IMCI, is to identify the treatable or preventable causes of non-malaria febrile illness .Antimalarial drugs are widely overprescribed: introduction of rapid diagnostic tests for malaria reduces overprescription and helps target antimalarial drugs to those who need themAntibiotics are also widely overprescribed, and antimicrobial resistance poses a fundamental threat to human health, development, and securityAt the same time as reducing overuse of antimalarial drugs, introduction of rapid diagnostic tests for malaria is associated with markedly increased levels of antibiotic prescribing especially for patients with negative test resultsThis effect is seen across multiple settings in Africa and Asia and, in large part, probably represents increased overprescribing of antibiotics


In this paper, concrete specimens are immersed in ultrapure water, to study the evolutions of surface permeability, pore structure and paste microstructure following the prolonging of immersion period. According to the results, after 30-day immersion, the surface permeability of concrete becomes higher as compared with the value before immersion. However, further immersion makes the surface permeability decrease, so that the value measured after 150-day immersion is only half that measured after 30-day immersion. The early increase in surface permeability should be mainly attributed to the leaching of calcium hydroxide, while the later decrease to the refinement of pore structure due to hydration. The two effects work simultaneously and compete throughout the immersion period. The proposed mechanisms get support from microscopic measurements and observations. This will undoubtedly increase the porosity [Concrete is one of the most widely used construction materials in civil engineering. It is also the main material in hydraulic engineering, due to its excellent resistance to water permeation. When concrete contacts with water, the calcium hydroxide in hydrated cement paste (the binder phase in concrete) will be leached out ,2. Leachporosity , and lowet al. [et al. [4NO3 solution after 19 days is 4–5 times higher than that in pure water after 114 days, and this is valid under both room temperature and high temperature (85 °C). Beside external surroundings, the leaching process is also affected by the microstructure of concrete. According to Maltais et al. [w/c) plays the most important role in controlling leaching. Smaller w/c results in denser microstructure, and improves the leaching resistance of concrete. When w/c is kept constant, the higher the aggregate volume fraction is, the lower the leaching rate will be.Leaching is the dissolution and diffusion of calcium ions in cement paste in nature. It has been considered as one of the reasons that induce durability problems, and investigated extensively. As an intrinsic problem of concrete when immersed in water, leaching has been studied by a number of researchers –7. Effecet al. , who fou [et al.  revealedperature 5 °C. BesAnother process simultaneous with leaching is the hydration of cement in concrete, which makes the microstructure of concrete denser and improves the impermeability of concrete . Under iEnvironmental loads always affect the surface layer of concrete first to form a micro-environment of a single kind of corrosive ions or multiple corrosive substances, then these substances can transport into the interior of concrete ,12. The 2.2.1.2, Al2O3, Fe2O3, MgO, SO3 and K2O take mass percentages of 64.67%, 18.59%, 4.62%, 4.17%, 2.35%, 3.32% and 0.92%, respectively. The loss on ignition of the cement is 1.03%. Other materials involved in the experiments include ultrapure water with the electrical resistivity of 18.2 MΩ·cm at 25 °C, river sand with the fineness modulus of 2.61 and the apparent density of 2632 kg/m3 as fine aggregate, and gravel with the particle size of 5–20 mm and the apparent density of 2700 kg/m3 as coarse aggregate.A CEM I Portland cement supplied by Shenzhen Haixing Onoda Cement Co. Ltd.  was used as cementitious materials in the preparation of concrete mixture. In the cement, CaO, SiO2.2.3 of fresh concrete are 175, 461, 720 and 1079 kg, respectively. After being mixed, the mixture was casted in steel moulds with the dimensions of 100 × 100 × 100 mm3, and covered with plastic sheets. After 24 h, all specimens were demoulded and cured in a moisture room where the temperature and relative humidity were 20 ± 2 °C and >95% respectively. At the age of 30 days, a part of the specimens were taken out to test for permeability, while the other specimens were immersed in ultrapure water, with the water surface 13 mm higher than the upper surface of the specimens. The volume ratio of liquid to solid specimens is approximately 3 for the immersions, and no refreshment of the liquid was made during the immersion period. Permeability values were also tested after the immersion durations of 30, 60, 90, 120 and 150 days.Fresh concrete mixture was mixed according to a mix proportion in which the water-to-cement ratio equals 0.38, and the masses of water, cement, sand and gravel in 1 mversus time. The Autoclam test has been widely used to assess the permeability and other properties of normal concrete and surface treated concrete [−7 m3/min0.5, and three parallel samples were tested and averaged to get one surface permeability index.Surface permeability was evaluated by a surface permeability index obtained from the Autoclam permeation testing equipment developed in Queen’s University Belfast, Belfast, UK. This test basically measures the volume of water penetrated in the concrete surface layer of a few millimeters, under pressure. The permeability index can be obtained from the curve of penetrated water concrete –17. At tt-plot, the famous BJH method [MIP (mercury intrusion porosimetry) and nitrogen adsorption were employed to characterize the changing of the pore structure of concrete following increasing immersion ages. For MIP tests, the surface layer (within 3 mm from the surface) of concrete specimens, both before and after immersion, were sawed off and crushed, and after the elimination of large aggregate particles, small pieces were taken as samples. The samples for MIP test were dried following the solvent (ethanol) replacement drying procedure suggested by Aligizaki . Based oH method  was empl3.3.1.−7 m3/min0.5. After immersed in ultrapure water, values of surface permeability index reach 1.296 ×10−7, 1.075 × 10−7, 1.019 × 10−7, 0.951 × 10−7 and 0.710 × 10−7 m3/min0.5 at the immersion ages of 30, 60, 90, 120 and 150 days, respectively. The determined surface permeability indices are plotted versus age (and immersion age) in Water seepage amounts recorded within the 15 min Autoclam tests on concrete at different immersion age are listed in 3.2.The pore size distribution (or cumulative porosity) curves of the concrete at the immersion ages of 0, 30 and 150 days, as measured by MIP, are shown in i.e., <100 nm, 100–200 nm and >200 nm, and named less effective pores, effective pores and very effective pores as referring to permeability. Pore size distributions of concrete specimens at different immersion ages expressed in this way are shown in Diamond, who introduced MIP into concrete technology in 1970s, has pointed out that the MIP pore size distribution curve is far from realistic, and MIP results are valuable only to provide intrudable porosities and threshold pore diameters which can serve as porosity index and connectivity index, respectively . In cont3.3.XRD patterns of concrete specimens at the immersion ages of 0 (when concrete is moisture-cured to the age of 30 days) and 30 days are shown in 4.In the present study, the most interesting finding is that, as shown in et al. [Since the ultrapure water was not refreshed during the immersion period, the concentration of calcium in the ultrapure water increased and the leaching rate decreased after a certain immersion period. According to Haga et al. , under tdc) can be used to assess the pore structure, i.e., smaller dc means finer pore structure. dc can be related to permeability through the well-known Katz-Thompson equation [Generally, in MIP results, a parameter named critical pore diameter After being immersed in water, calcium hydroxide will be leached from concrete, even though the concrete has been well cured.(2)After being immersed, surface permeability of concrete increases in the first month, and then drops down from a maximum value.(3)Leaching leads to higher porosity and coarser pore structure, while hydration makes the microstructure denser.(4)The evolution of the surface permeability of concrete is governed by the competition of leaching and cement hydration. When leaching is dominant in the first month of immersion, the surface permeability of concrete increases. Thereafter, when cement hydration rules, the surface permeability decreases.


Osteonecrosis of the jaw (MRONJ) linked to bisphosphonate treatment has specific characteristics that render its therapeutic management challenging for clinicians. Poor response to standard treatment makes it essential to take special precautions when treating this type of disease; therefore, antibiotic prophylaxis and/or antibiotic therapy have been proposed as effective and helpful tools in these situations.This article seeks to assess published evidence in order to evaluate the different protocols used for antibiotic prophylaxis and/or antibiotic therapy in the general context of patients treated with bisphosphonates.A literature review of the last 10 years was carried out in PubMed using the following keywords: “antibiotic prophylaxis and osteonecrosis,” “bisphosphonates AND osteonecrosis AND dental management,” “bisphosphonate AND osteonecrosis AND antibiotic prophylaxis AND oral surgery.” A total of 188 articles were obtained, of which 18 were ultimately selected.In patients treated with oral and intravenous bisphosphonates without chemotherapy-associated osteonecrosis of the jaw, antibiotic prophylaxis prior to oral surgery is an important tool to avoid osteonecrosis and promote healing of the affected area. If the patient previously exhibited chemotherapy-associated osteonecrosis after tooth extraction, antibiotic prophylaxis is indicated to prevent recurrent osteonecrosis and promote healing of the extraction site. If chemotherapy-associated osteonecrosis is already present, antibiotic therapy is a vital part of conservative management to reduce the symptomatology of MRONJ and keep it from worsening. Finally, a lack of clinical data and randomized controlled trials makes it difficult to choose the most appropriate protocol for the various clinical situations studied. Key words:Bisphosphonates, antibiotic prophylaxis, maxillary osteonecrosis, antibiotic treatment. Antibiotic prophylaxis is used in dentistry to prevent infections in high-risk cases, such as during surgical procedures that enable pathogens to enter the body, or in patients whose general health characteristics or specific medical condition make them more susceptible to contracting new infections -5.Consequently, prophylaxis is used to promote appropriate bioavailability of an antibiotic that can effectively tackle microorganisms and therefore prevent their proliferation and any subsequent infections. This concept is the opposite of antibiotic therapy, which is prescribed in cases of already established infection and is aimed at treating symptoms rather than preventing them .Antibiotic prophylaxis and/or antibiotic therapy protocols have been established as an effective therapeutic tool in the prevention or conservative management of certain diseases. One such disease is chemotherapy-associated osteonecrosis of the jaw caused by bisphosphonates (or other drugs), described by Marx in 2003  as an exA distinction should be made between oral and intravenous routes of administration of bisphosphonates ,12. IntrThe onset of bisphosphonate-related MRONJ has also been correlated with different local factors  and systemic factors  ,14.In view of these risk factors, patient medical history, and the oral-systemic regions affected by bisphosphonates , there are many reasons to use antibiotic prophylaxis in patients treated with oral or intravenous bisphosphonates.On the other hand, patients treated with oral or intravenous bisphosphonates who have already developed chemothera-py-associated osteonecrosis of the jaw may also benefit from antibiotic therapies to avoid potentially serious infection that can worsen symptoms; thus, antibiotic therapy can help improve overall clinical condition -17.The present article seeks to identify the different published protocols of antibiotic prophylaxis and/or antibiotic therapy in patients treated with oral or intravenous bisphosphonates. To this end, a systematic review of the literature was carried out focusing on three clinical situations: a) patients treated with oral or intravenous bisphosphonates and without MRONJ who will undergo a dental extraction, b) patients treated with oral or intravenous bisphosphonates and with previous incidence of MRONJ who will undergo a dental extraction; and c) patients treated with oral or intravenous bisphosphonates and with MRONJ, as part of their conservative management.A review of the literature published in PubMed over the last 10 years was carried out using the following keywords: “antibiotic prophylaxis and osteonecrosis,” “bisphosphonates AND osteonecrosis AND dental management,” “bisphosphonate AND osteonecrosis AND antibiotic prophylaxis AND oral surgery.” The results returned 29, 129, and 13 articles, respectively. The inclusion and exclusion criteria were then applied in order to fulfill the two proposed objectives.The inclusion criteria for the first proposed objective (antibiotic prophylaxis protocols) were: academic publications written in English that involved reviews of the literature, clinical trials, case control studies, cohorts studies and case series centered on antibiotic prophylaxis  in patients treated with oral or intra-venous bisphosphonates, with or without antecedents of developing chemotherapy-associated osteonecrosis of the jaw, who were undergoing oral surgery.The inclusion criteria for the second proposed objective (antibiotic treatment guidelines) were: academic publications written in English that involved reviews of the literature, clinical trials, or case series that specify the type of antibiotic used, doses and length of treatment, and guidelines used for antibiotic therapy in patients treated with oral and intravenous bisphosphonates with MRONJ, as part of their conservative management .The exclusion criteria for both proposed objectives were: articles that did not meet the eligibility requirements for both proposed objectives, articles unrelated to the research topic, articles that did not have an abstract or with an anonymous author, letters to the editor, and expert opinions.The selection process was finished by manually searching through all the bibliographic references of the selected articles.The initial search in PubMed yielded 171 results, with an additional 17 articles identified during the manual search of the collected articles’ bibliographic references. The flow chart seen in figure Results taken from the different articles can be seen in When antibiotic prophylaxis is used in patients without previous MRONJ who are receiving treatment with oral bisphosphonates , as wellIn the same way, the most widely used antibiotics for the treatment of MRONJ in patients taking oral bisphosphonates are penicillin, amoxicillin, amoxicillin/clavulanic acid, metronidazole, and/or a combination thereof.With regard to the length of prophylactic antibiotic treatment prior to and following tooth extraction, there is no uniform approach applied in all patients receiving treatment with oral or intravenous bisphosphonates. Despite this, most authors agree that post-extraction treatment regimens in patients receiving oral and intravenous bisphosphonates should be continued until the surgical site has completely healed.Antibiotic prophylaxis can be beneficial in avoiding the onset of MRONJ in patients who are set to undergo oral surgery (extraction) and are currently being treated with oral and intravenous bisphosphonates. If MRONJ has already developed and is under control, antibiotic treatment prophylaxis can prevent its recurrence. Moreover, antibiotic prophylaxis can help reduce the symptoms of osteonecrosis of the jaw, aiding in a conservative management regimen. However, despite the many studies found in the literature, there is no consensus on which is the most used antibiotic and its dosage .The latest consensus of the American Association of Oral and Maxillofacial Surgeons (AAOMS)  refers tAntibiotics used in patients without previous MRONJ who are receiving treatment with oral bisphosphonates, as well as in patients treated with intravenous bisphosphonates , have alFor patients receiving treatment with intravenous bisphosphonates, the articles’ recommendations for when to begin pre-extraction prescription of antibiotics range from one, ,19 two, For the above reasons, the total length of treatment time can vary greatly. Patients treated with oral bisphosphonates can receive prophylaxis for anywhere from seven  to twentRegarding the use of antibiotics for the treatment of MRONJ, the consensus of the AAOMS  identifiLong-term antibiotic treatment appears to yield better results in regard to promoting proper healing of the alveolar sockets ,20 and aThere is no consensus on total treatment time. Authors’ recommendations for conservative management of MRONJ using antibiotics include one week,  ten daysIn patients treated with intravenous bisphosphonates, the most commonly used antibiotics are penicillin, amoxicillin, amoxicillin/clavulanic acid, metronidazole, and/or a combination thereof. Some studies also prescribed penicillin G + IV metronidazole,  levofloxNo consensus exists on total treatment times, with the approach being similar to treatments described for patients undergoing treatment with oral bisphosphonates.In any case, it is always best to carry out antibiogram before prescribing any antibiotics;  however,Sparse clinical data and a lack of randomized controlled trials make it impossible to definitively identify the most appropriate protocol for each of the different clinical situations studied ,34.In conclusion, it is clear that in patients being treated with oral and intravenous bisphosphonates who have not had prior chemotherapy-associated osteonecrosis of the jaw, the use of antibiotic prophylaxis prior to oral surgery is an important tool in avoiding osteonecrosis and in promoting proper healing of the affected area. If a patient previously had chemotherapy-associated osteonecrosis after a tooth extraction, antibiotic prophylaxis will be indicated to prevent the recurrence of osteonecrosis and to promote healing of the extraction site. If chemotherapy-associated osteonecrosis is already present, antibiotic therapy is a vital part of conservative management to reduce the symptomatology of MRONJ and keep it from worsening.


The antibiotic name amoxicillin appears incorrectly throughout the article. The correct antibiotic name is amoxicillin-clavulanate.The antibiotic name amoxicillin appears incorrectly in Tables The antibiotic name amoxicillin appears incorrectly in


Control variables were: age, skin color, economic level, maternal education, physical activity and sexual maturation. Multinomial logistic regression was used for associations between the dependent and independent variables. Binary logistic regression was performed to identify the association between adolescents with all anthropometric indicators in excess and independent variables.The study included 1,132 adolescents (aged 14–19 years) enrolled in public schools of São José, Santa Catarina, Brazil, in the 2014 academic year. The dependent variable was the cluster of anthropometric indicators  of excess body fat. The independent variable was maximum oxygen uptake (VO2max unit, the odds of adolescents having three, four, five or more anthropometric indicators of excess body fat decreased by 0.92, 0.85 and 0.73 times, respectively. In the binary regression, this fact was reconfirmed, demonstrating that with each increase of one VO2max unit, the odds of adolescents having simultaneously the eight anthropometric indicators of excess body fat decreased by 0.55.One in ten adolescents presented all anthropometric indicators of excess body fat. Multinomial regression showed that with each increase of one VO2max unit, adolescents decreased the odds of simultaneously presenting three or more anthropometric indicators of excess body fat, regardless of biological, economic and lifestyle factors. In addition, the present study identified that one in ten adolescents had all anthropometric indicators of excess body fat.It was concluded that with each increase of one VO Anthropometry is one of the most widely used ways to evaluate body composition in epidemiological studies, since it is considered to be easy to apply, low cost and with good indexes of validity when compared to more precise methods . In this2max) and the greater amount of muscle mass is associated with higher VO2max values [2max levels is unclear. From the analysis of the relationship between body fat distribution and VO2max, it is possible to infer how fat localization interferes with performance in aerobic fitness tests, helping in the elaboration of strategies for maintenance and increase of VO2max levels [In addition to body composition being a strong indicator of health in adolescence, aerobic fitness is another component with good predictive health capacity in all life cycles . The litx values ,4. Howev2max, which justifies this study.Previous studies have analyzed clusters of some anthropometric indicators  . However2max values, increases the chances of adolescents presenting insulin resistance, metabolic syndrome and consequently higher morbidity rates and early mortality [2max are considered modifiable factors, which allow the development of actions related to increases in VO2max values, considering that VO2max is able to significantly reduce fat tissue [The grouping of anthropometric indicators demonstrates the simultaneity of health risk conditions related to the excessive amount of fat in different parts of the body, and this fact, added to the lower VOortality . Moreovet tissue .2max levels in adolescents.Therefore, the aim of the present study was to evaluate different clusters of anthropometric indicators  associated with VOThis cross-sectional study was approved by the Human Research Ethics Committee of the Federal University of Santa Catarina (UFSC) under CAAE protocol: 33210414.3.0000.0121.The study population consisted of 5,182 high school students aged 14–19 years enrolled in public high schools of the city of São José, southern Brazil, distributed in 11 eligible schools and 170 high school classes.The sampling process was determined in two stages: 1) stratified by public high schools (n = 11); 2) clusters of classes considering shift and school year (n = 170). In the second stage, all high school students who were in the classroom on the days of data collection were invited to participate in the research. The probabilistic sample consisted of 1,132 adolescents. Data on estimates for the calculation of the sampling process  can be found in literature . CollectThe dependent variable was the simultaneity of anthropometric indicators of excess body fat. Eight anthropometric indicators of body fat were analyzed: BMI, WC, WHtR, TR SF, SE SF, SI SF, ΣTR + SE and ΣTR + SE + SI.Anthropometric data on body mass, height, WC, and skinfolds were measured according to International Society for the Advancement of Kinanthropometry (ISAK) procedures by a single level 1 ISAK-certified assessor. For the measurements of skinfolds, Cescorf adipometer  with precision of 0.1mm was used.WC was measured with an inelastic Sanny metallic anthropometric tape with 0.1 mm resolution , measured at the lowest perimeter between the last rib and the superior border of the iliac crest. WHtR was evaluated by the relationship between WC (cm) and height (cm). BMI was calculated by the relationship between body mass (kilograms) and height in squared meters.th percentile of the sample distribution, considering that values above the 75th percentile are sensitive in the body fat diagnosis in children and adolescents [th percentile (P≥75) of anthropometric indicators of body fat were considered high because they represented the quartile with the highest percentage and for the cutoff points of all the anthropometric indicators to be equated. Score ranging from 0 (no anthropometric indicators of excess) to 8 (simultaneous presence of eight anthropometric indicators of excess) was used to classify individuals as for the simultaneity of anthropometric indicators.In the present study, all anthropometric indicators were classified as having as cutoff value the 75Aerobic fitness was estimated using the modified Canadian Aerobic Fitness test—mCAFT , validat2max of adolescents [Oxygen expenditure during exercise performed by the adolescent and reference values for the determination of the beneficial health zone for aerobic fitness are determined by the Canadian battery . The equlescents , which wSociodemographic variables were collected through a self-administered questionnaire. Skin color was self-reported according to the Brazilian Institute of Geography and Statistics  and dichPhysical activity was analyzed by frequency of practice in the last seven days, for at least 60 minutes. Adolescents who practiced less than five times a week were considered little physically active. This parameter was used from evidence that demonstrated that it is necessary to practice 60 minutes of physical activity in at least five times a week for health maintenance in adolescence .Sexual maturation was evaluated according to criteria proposed by Tanner , validatIn the descriptive analysis of variables, means, standard deviations and frequency distribution were used. Data normality of continuous variables was verified by analyzing asymmetry and kurtosis. Considering that all kurtosis and asymmetry values were close to zero, data were considered normal ,16. The The chi-square test of heterogeneity and linear tendency was used to evaluate differences between groups for each anthropometric indicator of body fat. The combinations among anthropometric indicators were presented and the ratio between observed and expected prevalence (O / E) was calculated . The obsThe associations between the dependent variable "simultaneity of anthropometric indicators of excess body fat" and independent variables were analyzed by multinomial logistic regression, with estimates of odds ratio (OR) and confidence intervals (95% CI). The reference category was "zero anthropometric indicator of excess body fat". Adolescents who presented six (3.8%), seven (5.1%) and eight (12.0%) anthropometric indicators of excess body fat were classified in the category "five or more anthropometric indicators of excess body fat" due to the low frequency.Binary logistic regression was performed with estimation OR and respective 95% CI to identify the association between adolescents who presented "eight anthropometric indicators of excess body fat" and independent variables. The reference category was "zero anthropometric indicator of excess body fat". The adjusted analysis was controlled by all independent variables. For all statistical tests, p <0.05 was used and the SPSS 21.0 software was used.2max of 38.80 ± 5.83mL / kg / min were analyzed , prevalence of approximately 267 times greater than expected was observed. When WHtR, TR SF, SE SF, SI SF, ΣTR + SE and ΣTR + SE + SI indicators were combined, the observed prevalence was 31 times higher than expected (0.032%). In relation to five anthropometric indicators , the prevalence observed was 22 times higher than expected. For the four anthropometric indicators, the prevalence of the combination of TR SF, SE SF, ΣTR + SE and ΣTR + SE + SI presented a higher O / E ratio (5.263%). For three anthropometric indicators , the observed prevalence was 2.827 times greater than expected. The simultaneity between BMI and WHtR had higher observed prevalence (0.861 times greater than expected). The highest observed prevalence for one anthropometric indicator in excess (WHtR) was 0.9% higher than expected .2max, the odds of adolescents having three, four, five or more anthropometric indicators of excess body fat decreased by 0.92, 0.85, and 0.73 times, respectively. Females were less likely of having three (OR = 0.39), four (OR = 0.17) and five or more (OR = 0.15) anthropometric indicators of excess body fat. In addition, adolescents with black / brown / yellow / indigenous skin color were twice as likely (OR = 2.28) of having simultaneity of four anthropometric indicators of excess body fat. Post-pubertal adolescents were more likely of having two (OR = 2.07) and five or more (OR = 1.70) simultaneous anthropometric indicators of excess body fat compared to the reference category (no anthropometric indicator of excess body fat index)  . Informat index) .2max increase, the odds of adolescents having the eight anthropometric indicators in excess were reduced by 0.55 times  anthropometric indicators of excess body fat. Other studies have shown that VO2max is inversely associated with body fat  and other cardiovascular risk factors in children and adolescents [2max on the inflammatory process caused by excess body fat.This study showed that the higher the VOlescents ,20. ThisExcess body fat is important inducer of systemic inflammation and this fact contributes to cardiovascular diseases linked to obesity . IncreasAs a consequence, CRP levels are closely linked to aerobic fitness levels . A study2max and body fat, investigated by clusters of anthropometric indicators, was maintained. This is in agreement with literature, which shows that regardless of any factors, low aerobic fitness is a risk factor for increased body fat levels and other health risk factors [This study used biological , economic  and lifestyle aspects  as control variables. Even with these variables, the inverse relationship between VO factors .As a complementary result of this study, it was found that 10.1% of adolescents had only one anthropometric indicator above normal, 14% had two to four indicators above normal and 24.6% had five or more indicators above normal . A study carried out with 627 Brazilian adolescents aged 14–17 years found a prevalence of 24.1% for the simultaneity of four anthropometric indicators at high levels  ,20,28. I2max), it is possible to elaborate VO2max improvement actions, predicting the changes that will occur in the different patterns of body fat distribution [Studies that use the grouping of anthropometric indicators can help in the control of errors due to the low sensitivity of the method (such as BMI), and consequently attenuate the underestimation of these results . More spribution . These dribution .2max was estimated by means of a submaximal test may be a study limitation, considering that the use of submaximal protocols to estimate VO2max has less precision than maximum protocols. However, submaximal tests are more practical to apply in samples with greater number of individuals [2max in adolescents with low physical fitness or who do not support maximal effort tests [The fact that VOividuals . In addirt tests .2max unit of adolescents increased, the chances of simultaneously presenting three or more anthropometric indicators of excess body fat decreased, regardless of biological, economic and lifestyle factors. In addition, the present study identified that one in ten adolescents had all anthropometric indicators of excess body fat.It could be concluded that as one VOS1 Tablea Reference category: zero anthropometric indicator of excess body fat. b Adjusted for all independent variables.OR, Odds Ratio; CI, Confidence Interval; M: mean; SD: standard deviation. (DOCX)Click here for additional data file.S2 TableOR, Odds Ratio; CI, Confidence Interval.(DOCX)Click here for additional data file.S1 FileORDER: Order Of StudentsID_QUIZ: Number of identification question in quizID_SCHOOL: Number of identification schoolSEX: female (number 2); male (number 3)AGE: yearsAGE_CATEGORIZED: 17–19 years (number 2); 14–16 years (number 3)SKIN COLOR: brown / black / yellow / indigenous (number 2); White (number 3)SCHOOL OF MOTHER: GREATER THAN 8 YEARS OF STUDY (number 2); LESS THAN 8 YEARS OF STUDY (number 3)ECONOMIC STATUS: low class (number 2); high class (number 3)ACTIVITY PHYSICAL: low active physically (number 2); physically active (number 3)SEXUAL MATURATION: post puberal/puberal (number 2); pre puberal (number 3)SUM 2: sum of triceps and subscapular foldsSUM 3: Sum of triceps and subscapular and suprailiaca foldsBODY MASS INDEX: overweight (number 1); normal weight (number 0)WAIST: normal (number 0); excesso (number 1)TRICEPS: normal (number 0); excesso (number 1)SUBESCAPULAR: normal (number 0); excesso (number 1)SUPRAILIACA: normal (number 0); excesso (number 1)SUM_2: normal (number 0); excesso (number 1)SUM_3: normal (number 0); excesso (number 1)SIMULTANEOUS BEHAVIOUR: number of concurrent behaviorsSTANDARD OF SIMULTANEITY: NUMBER OF EACH SIMULTANEOUS BEHAVIOURBEHAVIOUR FIVE OR MORE: ADOLESCENTS: 0 excess anthropometric indicator (number 0); 1 excess anthropometric indicator (number 1); 2 excess anthropometric indicators (number 2); 3 excess anthropometric indicators (number 3); 4 excess anthropometric indicators (number 4); 5 or more excess anthropometric indicators (number 5)BINARY REGRESSION: 0 to 7 anthropometric indicators in excess (number 0); 8 anthropometric indicators in excess (number 1).(XLSX)Click here for additional data file.