FA), silica fume (SF), ground granulated blast furnace slag (GGBS), metakaolin (MK), and rice husk ash (RHA). Comparison of normal and high-strength concrete in which cement has been partially supplemented by mineral admixture has been considered. It has been concluded that mineral admixtures may be categorized into two groups: chemically active mineral admixtures and microfiller mineral admixtures. Chemically active mineral admixtures decrease workability and setting time of concrete but increase the heat of hydration and reactivity. On the other hand, microfiller mineral admixtures increase workability and setting time of concrete but decrease the heat of hydration and reactivity. In general, small particle size and higher specific surface area of mineral admixture are favourable to produce highly dense and impermeable concrete; however, they cause low workability and demand more water which may be offset by adding effective superplasticizer.This paper presents a review of the properties of fresh concrete including workability, heat of hydration, setting time, bleeding, and reactivity by using mineral admixtures fly ash ( FA), silica fume (SF), ground granulated blast furnace slag (GGBS), metakaolin (MK), and rice husk ash (RHA) which possess certain characteristics through which they influence the properties of concrete differently. The reported benefits of mineral admixtures are often associated with the harden properties of concrete; however, mineral admixtures may also influence the properties of wet concrete between the time of mixing and hardening in one or more of the following ways such as they may affect water demand, heat of hydration, setting time, bleeding, and reactivity. In the authors' view, there is no literature that summarizes the effect of these mineral admixtures on the properties of fresh concrete. Moreover, effect of mineral admixtures on the durability and on the mechanical properties of concrete remained a focus of interest. Nevertheless, effect of mineral admixtures on the properties of fresh concrete is very important as these properties may affect the durability and mechanical properties of concrete. Comparative studies have been done such as the effect of blast furnace slag and fly ash on the hydration of fresh cement paste [SF), metakaolin (MK), fly ash (FA), and ground granulated blast furnace slag (GGBS) on the setting times of high-strength concrete [FA), silica fume (SF), ground granulated blast furnace slag (GGBS), metakaolin (MK), and rice husk ash (RHA) on the fresh concrete has been reviewed.Admixtures are added in concrete to improve the quality of concrete. Mineral admixtures include fly ash  from coal is a pozzolan, which results in low-permeable and more durable concrete, which is able to resist the ingress of deleterious chemicals. This pozzolan was first reported as a controller of damaging alkali-silica reaction (ASR) in 1949 by Blanks [FA concrete are usually higher than the concrete without FA which confirms that FA plays a preventive role against ASR that is an added advantage of FA [FA concrete. These structures were inspected in 1991 after about 35–40 years of their construction. The conditional survey, petrographic examination, and damage rating index (DRI) showed no symptoms of ASR [FA [McMillan and powers were the first who used coal nditions . FA or Py Blanks . Alkaliss of ASR . In 2010GGBS was first discovered in Germany by Emil Langen; however, commercial production of lime-activated GGBS was started in 1865 in Germany [GGBS has been named in the literature as slag concrete, which is being used successfully in many countries due to accepted benefits of this material and the use of slag concrete has been recommended in their national standards [GGBS was first used with Portland cement (PC). Since then it has been used extensively in many European countries. In the UK, the first British standard for Portland blast furnace cement (PBFC) was introduced in 1923 [In 1862, SF was first obtained in Norway, during filtration of the exhaust gases from furnaces as fumes. The large portion of these fumes contained very fine powder of high percentage of silicon dioxide. Since the 1970s, filtration of gases has started at large scale and, in 1976, first standard NS 3050 was granted to use SF in factory-produced cement, extensive literature is available on SF and SF concrete [SF of 8–10% by weight of cement is added in concrete, then its effect is between 50,000 and 100,000 microspheres percent particle; that is, concrete mix will be denser and cohesive due to fine particles of SF [In 1947, MK is a processed amorphous silica material and it is obtained from calcination of kaolin to a temperature between 600 and 850°C (1112 to 1562°F) [MK in concrete has no drawbacks; however, in 1995, Martin [MK in concrete increases the compressive strength up to 110 MPa (16 ksi) with superplasticizer to overcome the higher water requirement in MK concrete. In 1996, Wild et al. [MK is 20% by weight with superplasticizer of 2.4% of binder weight. Recently, Duan et al., 2013 [GGBS, SF, and MK and it has been found that MK has positive effects on pore structure and ITZ enhancement of concrete higher than SF and GGBS [ 1562°F) ; Larbi,  1562°F) ; Halliwe 1562°F)  reported, Martin  reportedd et al.  found thRHA has high pozzolanic activity due to noncrystalline silica and high specific surface area. RHA has been used in lime pozzolan mixes for partly replacement of Portland cement [Rice husk is the outer skin of the rice grain with a high concentration of silica, generally more than 80–85% . It occud cement –27.FA is produced when coal is burnt during power generation about 1600°C (2912°F) [2), alumina (Al2O3), iron oxide (Fe2O3), and other minor constituents. According to ASTM C 618-05, there are two classes of FA based on the types of coal from which it originates. The Class F is produced by burning anthracites which is mainly a siliceous and possesses pozzolanic characteristics. The Class C contains lime and higher MgO content and it is produced by burning lignite and subbituminous coal. Class C fly ash is lighter in colour in comparison with other ashes and may cause expansion and their strength behaviour at high temperature is not apparent [2912°F) . This bu12°F . Thapparent .GGBS contains silicates and alumino-silicates of calcium and is a by-product of iron manufactured in a blast furnace. There are two techniques, granulation and pelletization, through which GGBS is produced. In granulation technique, molten slag is forced over a weir into high pressure water jets which rapidly cools the slag as granules of 5 mm (0.197 in.) diameter; however, in pelletization technique, molten slag is poured into cold water rotating drum. The fins inside the rotating drum throw molten slag in the air towards the walls of the drum where water is sprayed to cool it rapidly. This process produces particles of size 100 mm (3.94 in.) to dust, out of which particle size of less than 6 mm (0.236 in.) in size is used to manufacture the GGBS. The materials produced from these two techniques can be used as raw material for glass structured GGBS [SF is a by-product obtained after reducing high-purity quartz with coal in electric arc furnace by heating up to 2000°C (3632°F) during the production of silicon. By oxidation and condensation of exhaust gas SiO, very fine spherical particles of SF are obtained which are highly reactive with the Ca(OH)2 produced during hydration of cement [SF consists of extremely fine particles of average diameters 0.1 μm (0.00000394 in.), having pozzolanic characteristics and tendency to develop plastic shrinkage cracks [f cement . SF conse cracks , 30.MK when it is heated to the temperature between 600 and 850°C (1112 to 1562°F) [MK is a very reactive pozzolan, but its physical and chemical characteristics greatly depend on the raw material used, the temperature during calcination and finishing process  [MK with a highly disorganized structure has been produced by normal cooling as well [Kaolin is converted into  as well , 11.RHA is produced by slow burning of rice husk at a temperature between 500 and 700°C (932 to 1292°F) [RHA may produce about 20% by weight of rice husk after incineration [Rice husk is produced in millions of tons per year as a waste material in agricultural and industrial processes.  1292°F) . RHA mayneration .FA reaction with ordinary Portland cement (OPC) is a two-stage process. In the first stage, during the early curing, the primary reaction is with CaOH2; however, the reaction rate depends on the curing temperature. At room temperature, the slower CaOH2 activation minimizes the reaction rate. The effectiveness of the use of fly ash in concrete depends on many factors including the following:FA and OPC;the chemical and phase composition of 2 concentration of the reaction system;Ca(OH)FA particles;the morphology of FA and OPC;the fineness of the development of heat during the early age of hydration process;FA.the reduction in mixing water requirements with FA affect early stage properties and the rheology of concrete [FA through trial mixes by considering workability, strength development, and durability [FA and produces additional hydration products of a fine pore structure.Variations in chemical composition and reactivity of concrete . It is arability . In the The pozzolanic reaction may be represented asFA concrete as compared to ordinary Portland cement concrete; however, despite this reduction, there is sufficient lime to maintain a high pH. It is important to mention that the resulting products due to the addition of FA are different from the resultant products formed in OPC concrete. FA produces very much finer pore structure with time presuming there is reached to water to maintain the hydration process [FA [Cabrera and Plowman (1987) showed that calcium hydroxide (lime) depletes with time and its reaction affects the long-term gain of strength in  process . A pH mocess [FA .FA in which Si–O–Si link has to break to make it reactive with lime, GGBS requires to be activated to react with lime. GGBS due to its glassy structure reacts very slowly with water in the presence of activators. Commonly, sulphates and/or alkalis act as activators, reacting chemically with the GGBS. These activators disturb the glassy structure and react to increase the pH of the system up to critical. In contrast to FA, GGBS only needs a pH level less than 12 and activators. In concrete, due to hydration of cement, Ca(OH)2 is produced and acts as an activator [Unlike ctivator .SF as a pozzolan reacts with Ca(OH)2 and about 25% of SF can consume most of the Ca(OH)2 at 28 days. This is very important as the Ca(OH)2 crystals are relatively weak, brittle, and not cementitious and cracks can easily propagate through regions concentrated with Ca(OH)2 crystals, that is, the aggregate cement paste matrix interface [nterface .MK [2·2Al2O3·2H2O into SiO2·2Al2O3 [MK of amorphous phase content less than 20% can be considered as inert materials in pozzolanic activity viewpoint [MK have also influenced the activity strength index ; however, there is no increase in activity strength index by the increase of amorphous phase index above 55% [The calcination process (dehydroxylation) of kaolin is actually a transformation from crystalline to amorphous phase. The amount and type of amorphous phase influence the activity of the additives . There aMK . Dehydro2·2Al2O3 . It is a2·2Al2O3 . MK of aiewpoint . AmorphoRHA and Ca(OH)2 solution yields C–S–H gel. The morphology of C–S–H gel is like congregate, having a large specific area due to the higher porous structure. This C–S–H gel and its large specific area are the main reason improving the concrete properties with the addition of RHA [Yu et al., 1999 , found tn of RHA , 35.The physical and chemical properties of mineral admixtures and OPC have been presented in RHA and SF showing their reaction capability with the primary hydrate of cement to produce calcium silicate hydrate (CSH) which is strengthening gel in concrete; however, the content of alumina oxide is lesser in SF and RHA. On the other hand, MK has substantial contents of silica and alumina oxide showing its capability to produce calcium silicate hydrate (CSH) and calcium aluminate hydrate (CAH) which has also bonding characteristics in the concrete.The most important constituents for any mineral admixture are silica and alumina oxides. In comparison with OPC, the mineral admixtures presented in RHA, SF, and MK have the smaller particle size and greater surface area showing their capability to react more effectively with Ca(OH)2 in the concrete.The particle size and surface area are the utmost important for mineral admixtures. As reported in the literature, smaller particle size with greater surface area is favourable within concrete to be more reactive with the alkaline environment . In TablFA, GGBS, SF, MK, and RHA through field emission scanning electron microscope (FESEM) has been shown in Figures GGBS and RHA have most uneven particle size distribution even though these FESEM images are taken after grinding. Hence, it may infer that grinding procedure and duration are very important in case of GGBS and RHA. GGBS, MK, and RHA have particles of irregular shape having multiple layer structure. Whereas, the FA and SF have spherical shape particles showing a possible increase in flow ability by packing of material once used in the concrete.The microstructure of FA can be related to water demand and fineness. For a specific workability, water demand can be reduced by varying particle shape and using finer fractions of FA [FA and found that the higher strength of FA concrete depends on reducing the water content and pozzolanic performance of the cement/FA combination. It is generally believed that finer FA significantly improves strength with time [FA of 45 μm (0.0018 in.) residue > 12 percent, the water requirements are greatly reduced [FA.According to Owens, 1979 , strengtns of FA . Dewar, ns of FA , correlaith time . Monk, 1 reduced . The fluFA, less water content is required when GGBS is added. Generally, 25% to 70% of cement is replaced with GGBS in the concrete [GGBS requires about 3% lesser water content in comparison to OPC for the equal slump requirement. This is due to the smooth surface texture of the slag particles that delay the chemical reaction and increase the setting time [Similar to concrete . GGBS reSF as cement replacing by mass increased the cohesiveness of concrete and required higher water content to maintain workability; however, through vibration or pumping, SF due to spherical particles gives concrete greater flow ability as compared to ordinary concrete [SF concrete without increasing the water/binder ratio, as it is important to use SF without strength loss. As a thumb rule, one part of SF can replace 3 to 4 parts of the cement by mass without loss of strength, provided the water content remains constant [concrete . The cohconstant .MK increases the cohesiveness of concrete [SF and MK is that both mineral admixtures have high reactivity and consume water very early.concrete , 43. Theconcrete . The reaRHA has to be used; this is also due to the high specific area of finer RHA which increases the water demand. The superplasticizer content has to increase more than 2.0% for RHA having a particle size of 11.5 μm (0.00045 in.) and specific surface of 30.4 m2/g (148533.6 ft2/lb.) [The superplasticizer content needs to be increased if finer and higher percentage of ft2/lb.) .FA, SF, MK, and RHA with different water/binder ratios. Zhang and Malhotra, 1996 [RHA and SF as cement replacing material. From their experimental results, the slumps of concrete having water/binder ratio 0.4, maximum binder content 386 kg/m3 (24 lb/ft3), and 10% cement replacement have been taken [RHA of different particle size (fineness) as cement replacing material. From their experimental results, the slumps of concrete having water/binder ratio 0.53, maximum binder content 391 kg/m3 (24.4 lb/ft3), and 20% cement replacement have been used [FA and SF as cement replacing material. From their experimental results, the slumps of concrete having water/binder ratio 0.56, maximum binder content 360 kg/m3 (22.45 lb/ft3), and 10, 15, 20, 30, and 40% cement replacement with the FA or a combination of FA and SF have been plotted [SF and MK as cement replacing material. From their experimental results, the slumps of concrete having water/binder ratio 0.23, 0.3 and 0.33; maximum binder content 500 kg/m3 (31.18 lb/ft3); and 5, 10, and 15% cement replacement with SF and MK have been shown [SF and MK as cement replacing material with fixed water/binder ratio of 0.35. From their experimental results, the slumps of concrete having maximum binder content 462 kg/m3 (28.82 lb/ft3) and 5, 10, and 15% cement replacement with SF and MK have been used [In ra, 1996 , investi86 kg/m3  lb/ft3, en taken , investieen used . Nochaiyeen used , investikg/m3 22. lb/ft3,  plotted , investikg/m3 22. lb/ft3, en shown , also inkg/m3 22. lb/ft3, SF and MK is 15% maximum, whereas FA and RHA have 30% and 20% maximum replacement levels, respectively. With respect to water/binder ratio, at 5% replacement level, the maximum slump is with MK concrete having water/binder ratio 0.33 (Wong and Razak series). Similarly, the maximum slump of SF concrete is with the water/binder ratio of 0.3 (Wong and Razak series). Further increase in water/binder ratio caused reduction in slump. Moreover, MK concrete has better slump as compared to SF concrete as shown in MK concrete but with water/binder ratio 0.3 (Wong and Razak series). The maximum slump of SF concrete at this replacement level is with water/binder ratio of 0.4 . At this replacement level, FA and RHA concretes have the same slump having water/binder ratio of 0.4  and 0.56 , respectively. At 15% replacement level the maximum slump is again with MK concrete but with water/binder ratio 0.3 (Wong and Razak series). The slump of SF concrete at this replacement level is extraordinarily reduced. It is also evident from MK concrete is less affected; however, SF concrete is very susceptible with the increase in replacement level specifically from 10% to 15%. The slump of concrete having combination of FA and SF as cement replacement has also been influenced by the addition of SF and the slump is further reduced with the increase in SF content. Though increase in FA content increase the slump as shown in SF on the slump are not fully canceled by FA. From RHA caused reduction in slump (Habeeb and Fayyadh series) [SF and MK require more water content due to small particle sizes, higher specific surface area and high reactivity.From  series) . In geneFA is effective in reducing heat of hydration. The exothermic reaction during the hydration of OPC liberates energy of 500 J/g (10455.8 lb. ft./oz). Use of FA by replacing some content of cement influences temperature rise during hydration process and increases pozzolanic reaction with increasing temperature; however, the peak temperature in FA concrete is significantly lower than equivalent ordinary Portland cement concrete.According to Woolley and Conlin, 1989 , and KecFA, as the proportion of GGBS is increased, the heat of hydration is reduced. This is beneficial in large concrete pouring that enable reduced temperature rise which will reduce the probability of thermal cracking [SF by using conduction calorimetry. By increasing the content of SF, higher heat of hydration has been observed. The rate of heat liberation, expressed on a cement basis, is increased as the amount of SF increases, but the total heat liberated, expressed on a total solid basis in the mixture, somewhat is decreased as SF is substituted for cement [Similar to cracking . Cheng-ycracking , have str cement .MK mortars relative to the plain OPC mortar showing an accelerating effect of MK on OPC hydration. Note that the maximum observed temperature rise occurs at 10% replacement of OPC by MK. This temperature rise has also been confirmed by Zhang and Malhotra, 1995 [MK with Ca(OH)2 is the cause of increase in temperature.Ambroise et al., 1994 , pointedra, 1995 . The higFA and GGBS, partial replacement of cement with RHA in concrete slows down the rate of hydration in comparison with ordinary concrete. The rate of hydration remains slow during the initial three days and consequently affects the strength after 150 days [2 supply to early heat evolution, by accelerating the hydration of OPC [2 [Similar to 150 days . In genen of OPC , 57 and f OPC [2 .FA depends on ambient temperature, cement content, fineness, water content, dosages of chemical admixtures, content of FA, fineness, and chemical composition of FA [FA from 10 different sources have been conducted to estimate water requirement, setting time, and bleeding [FA. No steady water reduction was observed in Class F FA concrete. Minor increase in setting time was observed in FA concretes [The setting time of concrete in the context of on of FA . Tests obleeding . The watoncretes .GGBS slowly reacts with water as compared to Portland cement, therefore stiffening/setting time of concrete is high. The setting time will be greater at high replacement levels above 50% and at lower temperatures (below 10°C (50°F)) [SF, having a greater surface area and higher silicon dioxide content, is found to be more reactive than Pulverized fly ash (PFA) or GGBS. The high reactivity increases hydration rate of the C3S fraction of the cement in the first instance and thus decreases the setting time [Since MK are increased and are extended further with the increase in replacement level; however, by increasing the replacement level up to 15%, minor drop in initial setting time particularly is observed in comparison with 10% MK concrete [MK at the higher replacement level. With higher MK content, concrete due to higher binder phase becomes denser and results in accelerating the setting [Initial and final setting times of concrete containing 10% concrete . This mi setting ; however setting .SF concrete, the initial and final setting time of RHA concrete has been observed and initial and final setting time of RHA concrete is about 29 and 60 minutes longer than SF concrete, respectively [SF and MK concrete as compared to plain concrete. This may be due to the fact that SF and MK accelerate the hydration of cement after initial set [In comparison to ectively . Hence, tial set . In genetial set . Thus, otial set .FA showed less bleeding than plain concrete. Also, concrete with Class C FA showed lesser bleeding than concretes with Class F FA. The reduction in bleeding is due to greater surface area of particles of fly ash and lower water content with fly ash for a given workability [SF or RHA produces no bleeding water [MK as a partial replacement of cement in suitably designed concrete mixes improves cohesion and reduces bleeding of fresh concrete [Gebler and Klieger, 1986 , reportekability . On the ng water . Similarconcrete .MK is higher than the other pozzolan. The authors conducted the same test on SF and MK as shown in MK in a cement matrix eliminated the calcium hydroxide completely. Nevertheless, MK reduces the calcium hydroxide level in concrete but pH remains stable above 12.5 [Reactivity of pozzolan may be compared with the help of Chapelle test. This test is performed by the reaction of calcium hydroxide with the dilute slurry of the pozzolan at a temperature of 95°C (203°F) for 18 hours. After reaction, the consumed calcium hydroxide is determined. Largent, 1978 , reporteove 12.5 .SF and MK are chemically active mineral admixtures, whereas FA, GGBS, and RHA are microfiller mineral admixtures. Based on these two groups, the following generalized conclusions can be drawn on the properties of fresh concrete.Chemically active mineral admixtures (highly reactive pozzolan) increase the cohesiveness of concrete and require more water to maintain workability; however, the requirement of water may be offset by adding plasticizer. The water demand depends on the particle size, specific surface area, particle shape, replacement level, and reactivity of particular mineral admixture.The workability of concrete with microfiller mineral admixtures (low to moderate reactive pozzolan) greatly depends on the particle size, specific surface area, particle shape, and replacement level. In general, smaller the particle size and higher the specific surface of mineral admixture increases the water demands of concrete. The low to moderate reactivity and filler effect help to maintain the workability and sometimes increase the workability.Heat of hydration increases with the use of chemically active mineral admixtures and decreases with the use of microfiller mineral admixtures.Initial and final setting time of concrete depends on water content, initial and curing temperature, dosage of superplasticizer, and the reactivity of mineral admixture. Thus, on similar conditions, initial and final setting times of concrete decrease with the use of chemically active mineral admixtures and increase with the use of microfiller mineral admixtures.In general, setting time of concrete decreases with the increase in replacement levels of chemically active mineral admixtures and increases with the increase of microfiller mineral admixtures.All mineral admixtures studied reduce bleeding in concrete with correct proportion of all ingredients.MK > SF > FA > GGBS.Based on Chapelle test, the reactivity of mineral admixtures is of the order: Smaller particle size and higher specific surface area of mineral admixtures are favourable to produce highly dense and impermeable concrete; however, they cause low workability and more water demand which may be offset by adding effective superplasticizer.Based on the review, it is quite clear that mineral admixtures may be categorized into two groups, namely, chemically active mineral admixtures (highly reactive pozzolan) and microfiller mineral admixtures (low to moderate reactive pozzolan).


Racetrack surface is a risk factor for racehorse injuries and fatalities. Current research indicates that race surface mechanical properties may be influenced by material composition, moisture content, temperature, and maintenance. Race surface mechanical testing in a controlled laboratory setting would allow for objective evaluation of dynamic properties of surface and factors that affect surface behavior.To develop a method for reconstruction of race surfaces in the laboratory and validate the method by comparison with racetrack measurements of dynamic surface properties.in situ (racetrack) and using laboratory reconstructions of harvested surface materials. Clegg Hammer in situ measurements were used to guide surface reconstruction in the laboratory. Dynamic surface properties were compared between in situ and laboratory settings. Relationships between racetrack TTD and Clegg Hammer measurements were analyzed using stepwise multiple linear regression.Track-testing device (TTD) impact tests were conducted to simulate equine hoof impact on dirt and synthetic race surfaces; tests were performed both Most dynamic surface property setting differences (racetrack-laboratory) were small relative to surface material type differences (dirt-synthetic). Clegg Hammer measurements were more strongly correlated with TTD measurements on the synthetic surface than the dirt surface. On the dirt surface, Clegg Hammer decelerations were negatively correlated with TTD forces.in situ values. The negative correlation between TTD and Clegg Hammer measurements confirms the importance of instrument mass when drawing conclusions from testing results. Lighter impact devices may be less appropriate for assessing dynamic surface properties compared to testing equipment designed to simulate hoof impact (TTD).Laboratory reconstruction of racetrack surfaces guided by Clegg Hammer measurements yielded TTD impact measurements similar to Dynamic impact properties of race surfaces can be evaluated in a laboratory setting, allowing for further study of factors affecting surface behavior under controlled conditions. Racetrack surface  is considered a risk factor for racehorse injuries and fatalities, but epidemiologic evidence, likely confounded by simultaneous risk factors, is contradictory providing no clear direction for desirable racetrack surface composition Initially, research examining race surfaces used subject-based (racehorse) methods, including observational studies in situ (racetrack) measurements is needed to establish the value of laboratory measurements. Validation would allow for future studies of surface behavior and associated environmental factors in a controlled, laboratory setting.Equine race surface dynamic properties have been evaluated in controlled laboratory settings in situ surface dynamic properties.The objectives of this study were to develop a method for laboratory reconstruction of equine race surfaces and to validate this method by comparison with in situ  and after reconstruction of harvested surface materials in a laboratory box (3 experiments per surface). Clegg Hammer deceleration measurements at the racetracks guided laboratory surface reconstruction. Surface horizontal properties were assessed using a shear vane tester. Laboratory and racetrack measurements were compared, as well as, the relationships between Clegg Hammer and TTD measurements.A track-testing device (TTD) The TTD g). Residual deformation, the resting deformation relative to the pre-impact surface height  was measured using a ruler (0.32 cm (0.125 in) resolution) attached to the Clegg Hammer handle.A 4.5 kg, 5 cm diameter cylindrical Clegg Hammer  was used to assess surface compaction. This device was portable, standardized 2). Force and torque data were collected at 10 Hz. Force and torque data were converted to normal stress (σ) and shear stress (τ) by dividing by the surface area of soil affected by the shear interface.A custom shear vane tester  of 83% sand, 10% silt, and 7% clay. The synthetic surface material was a proprietary wax-coated blend of sand (approximately 80% of composition), rubber, synthetic fibers and 15.7% organic matter. Surface material (∼380 L) was harvested in 4 layers by depth (2 inches each) at a single location from each racetrack for surface reconstruction in the laboratory.TTD and Clegg Hammer tests were conducted at 3–5 locations/day for 4 days on each racetrack, starting on the homestretch (near the finish line), then 2–4 backstretch locations, and when possible, ending at a new location on the homestretch. At each location, 2 TTD impacts  were performed at 6 spots spread over 7–10 m. The first 3 spots were vertical impacts with 20.3, 30.5, and 40.6 cm TTD drop distances, respectively; the next 3 spots were angled  impacts.Clegg measurements (5 consecutive 45 cm drops) were taken at 4 depths  per location at individual spots within 1–3 m of a TTD spot.Surface material temperature was measured with an infrared pen thermometer  at each test location, at each depth (4 layers). Moisture content surface material samples (100–250 g) were obtained from the uppermost layer at all locations, and from depths 5, 10, and 15 cm below the uppermost layer from at least one location per day. Moisture content was determined using a modification of an ASTM standard Shear vane tests were performed at 1–2 locations/day. For each test, the shear vane was placed at a new harrowed location, a weight was applied, and the shear vane was rotated approximately 100° in 30–60 seconds. The 3 weights  were each tested twice in random order at each location.width×length) wood laboratory box with a base layer designed for a synthetic surface Race surfaces were reconstructed in the laboratory track-in-a-box, a 101.6 cm×101.6 cm , a manual impact-driven custom compaction device , a 95 cmnd drop impact deceleration (CIV2) was within 10–20% of the racetrack average for each layer. CIV2 was chosen as the target value because the 1st drop (CIV1) was often a seating drop, and CIV1 was often immeasurable at the racetracks (4.5 kg Clegg Hammer cannot read CIV<2.6). Racetrack data regression analyses showed upper layers were more correlated to TTD values, compared to lower layers , the softer cushion layer was added to bring the surface height to 20.3 cm. Harrow methods were derived from knowledge of racetrack methods and equipment, and from preliminary testing results. Surfaces were harrowed by spring tines  with depThe laboratory portion of the study was designed as a split-plot ANOVA, where alpha = 0.05, SD of 0.05, and detectable contrast of 0.1 (twice as much as the SD). A t-test was used to compare laboratory and track differences. For each of the 3 experiments (reconstructions) for the dirt and synthetic surfaces, 2 sets of 6 TTD impacts  were performed in the same randomized order as performed at each racetrack. The upper portion of the surface was harrowed after every impact. A single 40.6 cm vertical impact was performed between sets to check the stability of the values.Shear vane tests were performed during the third experiment for each surface. For each test, the shear vane was placed on a harrowed location in the laboratory box and rotated approximately 110° in 90–150 seconds. The 3 weights  were each tested 4 times in random order for each surface reconstruction.Moisture content was measured for surface material samples from each layer  at the end of each experiment. For the dirt surface, moisture content was measured at 0 and 5 cm below the top of the surface at the beginning and middle of the experiments to verify that moisture content was maintained as moisture was added periodically to replace moisture loss to the environment.Maxima, minima, and other attributes of force, moment, and displacement were extracted from data using custom software . Zero force was established as average force over 0.1 s during free fall. A threshold vertical force of 40 N was used to determine times of impact initiation, termination, and duration. Impact was defined as the period of time when vertical force exceeded 40 N. Extracted data include impulse , time to maximum force (time from impact initiation to peak force), average load rate (maximum force divided by time to maximum force), average rebound rate (maximum force divided by the time from peak force to impact termination), average stiffness (maximum force divided by maximum deformation), maximum load rate , maximum rebound rate (minimum slope of force versus time) and maximum stiffness (maximum slope of force versus displacement).Zero deformation was established as the position at impact initiation Velocity was calculated from the time derivative of a quadratic fit of displacement versus time during free fall, and after impact. Impact and rebound velocities were the velocities at impact initiation and termination, respectively.mv2/2) and potential energy (mgh) at respective time points. The potential energy datum was defined as the height at maximum deformation. Energy returned to the TTD (%) was defined as energy at impact termination divided by energy at impact initiation.Total mechanical energy at impact initiation and termination were calculated as the sum of kinetic (max) and corresponding normal stress were fitted linearly to solve for cohesion (c) and angle of internal friction (φ) in the Mohr-Coulomb equation (τmax = c+σ tan φ) Force and torque data were converted to normal stress (σ) and shear stress (τ) by dividing by the surface area that was compressed and sheared, respectively, by the shear interface. Values of maximum shear stress  were used in the regression analysis. The Clegg Hammer cannot record impact values less than 2.6. Because of this, values were not recorded for some tests. For each surface, the first 2 Clegg Hammer impact decelerations without missing values were used in the analysis . BecauseThe stepwise procedure was first used to select possible Clegg Hammer predictor variables  related to individual TTD variables . Subsequst, 2nd), and all interactions significantly changed the magnitude of the setting difference. Finally, laboratory measurements were analyzed independently from in situ measurements using repeated measures mixed model ANOVA to assess the effect of surface, impact velocity, impact angle, set, and all interactions on TTD dynamic surface properties. Tests of the assumptions inherent in the statistical analyses were performed and met in the current study. For example, assessment of normality of the residuals from ANOVAs were examined. Those variables not exhibiting normality by the Shapiro-Wilk test were further assessed by histogram plots, which were determined to be normal.Laboratory and racetrack TTD measurements were compared using a one-sample t-test on the differences between the laboratory value and racetrack average for each surface in situ (racetrack) measurements.Following layered hardness surface material layer reconstruction, laboratory dynamic properties closely approximated Clegg Hammer-TTD correlations were stronger for the synthetic racetrack surface than for the dirt racetrack surface . Clegg HLaboratory dirt surface material moisture content measurements were slightly less (≤0.5%) than those at the racetrack except for the uppermost layer (0 cm), where the laboratory moisture content was 1.9% greater than the racetrack moisture content . LaboratLaboratory ambient and surface material temperatures were approximately 21.5°C. Ambient temperatures were 13.3±4.0°C at the dirt racetrack and 18.1±6.1°C at the synthetic racetrack. Racetrack surface material temperatures were warmer than the average ambient temperature, with upper layers warmer than lower layers .nd drop impact decelerations (CIV2) were 91–120% of racetrack averages  for the 3 drop distances  were 1.85±0.05, 2.25±0.04, and 2.58±0.06 m/s at the racetracks, and 1.82±0.05, 2.22±0.05 and 2.53±0.05 m/s in the laboratory. On average, laboratory impact velocity was 0.03 m/s (1.3%) less and significantly different than racetrack impact velocity (P<0.001). Angle impact velocity was always less than the corresponding vertical impact velocity, 2% less at the racetracks and 4% less in the laboratory.in-situ surface type differences (dirt-synthetic) (ΔSurfT), the setting differences were less than 14% for these same key values (T) was very small for time-related and energy-related variables . Consequently, the setting difference was a larger proportion of the racetrack surface type difference for these variables.Most TTD setting differences  were 10% or less of the respective racetrack averages, and the differences were less than 6% for key values like Z-force maximum, average load rate, and average stiffness . When coy values . The laby values . The diry values . The synSetting differences  were significantly affected, but to a relatively small degree, by surface type, impact velocity, and impact angle for 3, 10, and 19 parameters, respectively , synthetic shear failure stress was 5% greater at the laboratory than at the racetrack, whereas dirt surface shear failure stress was 27% greater. At a normal stress of 400 kN/m2, the laboratory dirt surface shear failure stress was 2% greater than the laboratory synthetic surface value.Shear stresses ranged from 10–29 kN/mboratory , and froin situ (racetrack) and in the laboratory. Laboratory surface reconstruction was guided by racetrack Clegg Hammer deceleration measurements. Relevance of observed differences in the dynamic behavior of race surfaces between settings was determined by comparison with observed differences between the dirt and the synthetic surfaces. Differences between racetrack and laboratory measurements were small relative to differences observed between dirt and synthetic surfaces . Thus, laboratory reconstruction of racetrack surfaces resulted in acceptable replication of measured dynamic impact properties of race surfaces for the desired application. However, due to the limited number of surface materials and maintenance procedures evaluated in the present protocol, findings in this study cannot be extended to all dirt and synthetic surfaces and their various maintenance conditions.The validity of measuring dynamic impact properties of racetrack surfaces in the laboratory was examined by comparing measured properties in situ surface type differences. Laboratory tests were developed to assess surface type differences in a controlled setting. The racetrack surface reconstruction protocol reproduced measurements sufficient for the intended application.Small differences were observed between laboratory and racetrack measurements for dynamic properties that may be related to racehorse injury in situ (racetrack).The effects of surface, impact velocity, and impact angle on dynamic surface properties were very similar in the laboratory to those observed at the racetrack Multiple impact tests can be performed on the same constructed surface in the laboratory with appropriate surface preparation. In the present study, measurements from two sets of impact tests (interspersed with harrowing) on each reconstructed laboratory surface were not significantly different between sets. This result coincides with previous studies Although Clegg Hammer impact deceleration measurements appeared useful for layered reconstruction of race surfaces in the laboratory, correlations between TTD and Clegg Hammer measurements at the racetracks were moderate to good for the synthetic surface and weak for the dirt surface. The better observed correlation for the synthetic surface may be related to lower variability across a range of impact velocities and two different impact masses . These findings are similar to TTD results from a previous study Upper layer hardness is affected by lower layers. Clegg Hammer measurements from more superficial layers  of the racetrack surface were more highly correlated with TTD dynamic surface properties than those for deeper layers. For this reason, the deepest layer at the laboratory (15 cm) was often made harder than the racetrack average to ensure that upper layers achieved the desired hardness. Note that the Clegg Hammer is purported to analyze up to about 15 cm in depth Laboratory TTD measurements more closely approximated dirt racetrack measurements, compared to synthetic. Both surfaces exhibited more plastic behavior than elastic behavior Moisture content discrepancies between laboratory and racetrack settings likely contributed to some differences in dynamic surface properties between settings. Dirt surface dynamic properties are affected by moisture content in situ.Setting differences in synthetic surface dynamic properties may be attributed to temperature differences. Synthetic surface properties are affected by temperature because their waxes undergo melting and softening at operating temperatures common to racetracks Several reconstruction factors likely affected replication of racetrack dynamic surface properties in the laboratory. During reconstruction, it was more difficult to achieve compaction of the synthetic surface because of its composition. The additives and composition of the synthetic surface resisted the settling of particles during vibration, and thus impeded compaction efforts. Overall, laboratory reconstruction and harrowing were simplified as surfaces were constructed to be level, not graded like the racetracks, and the speed and force of harrowing did not fully replicate harrowing performed by tractors. Furthermore, laboratory surfaces were built in one day. Racetrack surface behavior is the result of different combinations of surface harrowing, water application, environmental factors, and horse traffic that occur over much longer periods of time. These differences could have affected surface cushion hardness and density. The surface cushion in the laboratory was mostly harder than that at the racetracks as shown by upper layer (0 cm) Clegg measurements . More specifically, cushion hardness could have affected time of detection of the 40 N force threshold for determination of impact initiation, and thus the velocity at impact initiation. Initiation would have been detected earlier in free fall for a harder cushion, which would explain lower impact velocities observed in the laboratory. Better replication of the surface cushion layer could decrease differences in deformation and time-related measurements.Quasi-static shear vane measurements were not well replicated for the dirt surface. Simple racetrack measurements that capture the rate-dependent and horizontal surface properties in addition to the Clegg Hammer deceleration values may enhance the laboratory surface reconstruction guide used in the present study. Future studies should further explore horizontal properties of equine race surfaces.Findings in the current study are limited to the specific racetrack surfaces, materials and environmental conditions examined. The behaviors of materials and reconstructed surfaces are expected to be non-linear and highly affected by a multitude of conditions . The current study did not address reproducibility of other surfaces or alternative reconstruction techniques. However, the study did provide the first objective data on racetrack surface mechanical behavior within a scaled, controlled laboratory setting, as well as evidence for the potential usefulness of this approach in the evaluation of racetrack surfaces. This approach is particularly valuable when considering the $8–10 million cost of replacing a single racetrack surface in the US. Changing racetrack surfaces in order to empirically evaluate conditions is not economical or practical. Therefore, this study is considered an initial step in the development of methodology for evaluating the properties of racetrack surfaces as a structure composed of surface material layers.Important relationships between TTD and instrumented horse measurements have not yet been quantified. When horse hoof acceleration and TTD  measurements from a dirt surface were fitted with polynomial equations, nonlinear relationships were found that were dependent on the horse gallop speed in situ. The laboratory method used in the present study was useful for the objective evaluation of dynamic properties of racetrack surfaces in a controlled environment. Impact measurements derived from light, portable instruments may lead to different conclusions, compared to measurements from heavier devices designed to simulate a racehorse’s hoof impacting at the gallop.In summary, equine racetrack surfaces were successfully reconstructed in the laboratory by replicating racetrack Clegg Hammer deceleration measurements throughout the depth of the surface reconstruction. While exact replication of all properties was not achieved, setting differences observed between laboratory and racetrack values for dynamic surface properties central to equine injury prevention were small, particularly relative to differences between dirt and synthetic surfaces


Methods. We conducted a case-control study of 93 patients with recurrent pregnancy loss (cases) and 206 healthy multiparous women (controls) in a Colombian subpopulation. Three single nucleotide polymorphisms (SNPs) markers of the inherited thrombophilias factor V Leiden, prothrombin G20210A, and methylenetetrahydrofolate reductase C677T were genotyped by PCR-RFLP. Activated protein C resistance and plasma levels of antithrombin, protein C, and protein S were also measured. Results. The frequency of thrombophilia-associated SNPs, activated protein C resistance, and anticoagulant protein deficiencies, was low overall, except for the methylenetetrahydrofolate reductase C677T SNP. The differences between patients and controls had no statistical significance. Conclusion. Our study confirms the low prevalence of inherited thrombophilias in non-Caucasian populations and it is unlikely that the tested thrombophilias play a role in the pathogenesis of recurrent pregnancy loss in this Colombian population.Studies have shown an association between recurrent pregnancy loss and inherited thrombophilia in Caucasian populations, but there is insufficient knowledge concerning triethnic populations such as the Colombian. The aim of this study was to evaluate whether inherited thrombophilia is associated with recurrent pregnancy loss.  Pregnancy is an acquired hypercoagulable state and women with a prior tendency to thrombosis may develop clinical symptoms of placental vascular complications such as preeclampsia, intrauterine growth restriction, and fetal death for unknown causes, that impact the maternal-fetal morbidity and mortality –4. RecurMost of the studies on thrombophilia and recurrent pregnancy loss have been conducted in Caucasian populations , 22. TheAn unmatched case-control study was conducted on 93 patients and 206 healthy women from a province in Northwest Colombia (Antioquia) chosen for convenience. The inclusion criteria for the patient group were a history for recurrent pregnancy loss (three or more miscarriages) and any of the following vascular pregnancy manifestations: one or more second-trimester or later losses, severe or recurrent preeclampsia, intrauterine growth restriction, placental abruption, or otherwise unexplained intrauterine death. The women with recurrent pregnancy loss were selected among more than 3000 patients who had consulted, over the last 20 years, to the Reproduction Group at Universidad de Antioquia in Medellín, Colombia. The control group included 206 healthy women with a history of two or more normal pregnancies, no more than one miscarriage, and no history of any of the criteria for inclusion in the patient group. The sample size estimated for a type I error of 5% and a power of 80% was 300 subjects (2 : 1 ratio between controls and cases). This estimation was based on the following average ratios of SNPs in healthy European populations and their odds ratio for recurrent pregnancy loss: 4.3% of heterozygous FV Leiden with an odds ratio (OR) of 4 , 26; 3% Blood samples were drawn from women who were not taking any anticoagulant drugs and who were at least 3 months past the last obstetric event to prevent changes in the APC resistance status and plasma levels of coagulation proteins.a/b was lower than 2 being (a) in the presence, and (b) in the absence of activated PC. The activities of AT, PC, and PS were determined with commercial kits . These proteins were considered deficient when their activity was <50% of their normal functionality.Thrombophilia-associated SNPs were genotyped using the methods described by Ridker et al.  (FV Leidtamaño de la muestra (versión 1.1) [The sample size was calculated using the software ión 1.1) . The allión 1.1) . The ORsión 1.1) .Demographics of the women from the case and the control groups are shown in  The present study is the first to report the frequency of inherited thrombophilia  in a group of Colombian patients with recurrent pregnancy loss. Thrombophilia-associated SNPs such FV Leiden and FII G20210A were very low in both patients and controls, and although the frequency of MTHFR C677T SNPs was high in the patients, differences were not significant. Other inherited thrombophilias such as PC and AT deficiency are not commonly found in patients who had already experienced recurrent pregnancy loss. Only three patients with PC deficiency and no patients with AT deficiency were found among 93 cases. Overall, we found a low frequency of inherited thrombophilia in our patient population unlike Caucasian populations; therefore, it is unlikely that the thrombophilic alterations studied play a role in the pathogenesis of recurrent pregnancy loss in this Latin American population. n = 114) was evaluated with the following results in patients versus controls: 10% versus 0.9% were heterozygous for FV Leiden, 3% versus 0% were heterozygous for FII G20210A (one patient was homozygous), 24% versus 19.3% were homozygous for MTHFR 677T, and 25.2% versus 5.3% had APC resistance  [The low frequency of FV Leiden in our study agrees with two previous reports on the Colombian population. Camacho Vanegas et al. tested 150 healthy individuals from Bogota (30–50 years old) using PCR-RFLP and found no evidence of FV Leiden ; howeverrphisms) . During rphisms) . The FV Leiden, as genetic factor involved in the etiology of thrombosis, has motivated research of this abnormality as a risk factor for recurrent pregnancy loss, as reviewed by Krabbendam et al. ; howeverOur results are in contrast with previous recurrent pregnancy loss studies in Latin American populations. Brazilian women carrying FV Leiden had a 4.9 times higher risk of recurrent pregnancy loss than noncarriers . TherefoIn the current study, the wide confidence intervals suggest that the estimated sample size of 300 individuals was insufficient to detect differences between patients with recurrent pregnancy loss and control women for whom the power of the study decreased from an estimated 80% to 30% for detections of an OR of 2.0 with a frequency of 0.5% for FV Leiden in the controls. Nevertheless, our results are valuable because they confirm that the frequency of these inherited thrombophilias in our populations is low; therefore, we believe that there is no justification to recommend testing inherited thrombophilias in the initial evaluation of patients with recurrent pregnancy loss. To detect an association between thrombophilias and recurrent pregnancy loss, it would be necessary to increase considerably the sample size but with unlikely clinical significance.


Doxorubicin (DOX) is associated with premature cardiovascular events including myocardial infarction. This study was performed to determine if the weekly administration of DOX influenced coronary arteriolar medial and/or adventitial wall thickening.Thirty-two male Sprague-Dawley rats aged 25.1± 2.4 weeks were randomly divided into three groups and received weekly intraperitoneal injections of normal saline , or low  or high  doses of DOX. The animals were treated for 2–12 weeks, and euthanized at pre-specified intervals  to obtain histopathologic assessments of coronary arteriolar lumen diameter, medial wall thickness, adventitial wall thickness, and total wall thickness .p = 0.22). In comparison to animals receiving weekly saline, animals receiving weekly injections of 2.5 mg/kg of DOX experienced an increase in medial , and total wall thickness , respectively. These increases, as well as adventitial thickening became more prominent after normalizing for lumen diameter (p<0.05 to p<0.001) and after adjusting for age, weight, and total cumulative DOX dose (p = 0.02 to p = 0.01). Animals receiving low dose DOX trended toward increases in adventitial and total wall thickness after normalization to lumen diameter and accounting for age, weight, and total cumulative DOX dose .Lumen diameter was similar across all groups (saline: 315±34 µm, low DOX: 286±24 µm, high DOX: 242±27 µm; In conclusion, these data demonstrate that weekly treatment of rats with higher doses of DOX increases coronary arteriolar medial, adventitial, and total wall thickness. Future studies are warranted to determine if DOX related coronary arteriolar effects are reversible or preventable, exacerbate the known cardiomyopathic effects of DOX, influence altered resting or stress-induced myocardial perfusion, or contribute to the occurrence of myocardial infarction. Cardiovascular (CV) events including myocardial infarction (MI), stroke, and congestive heart failure (CHF) are the second leading cause of premature morbidity and mortality for those surviving beyond 5 years of their initial diagnosis for breast cancer or a hematologic malignancy such as Hodgkin's disease or Non-Hodgkin's lymphoma To this end, we investigated the association between the administration of DOX and histopathologic changes in coronary arteriolar microcirculatory arterial segments. To address this issue, we performed a series of studies to determine the relationship between DOX administration and measures of coronary arteriolar wall thickening. In this study, we evaluated 2 doses of DOX in a rat model without coronary arterial atherosclerosis. These doses were selected to mimic doses commonly administered to patients receiving DOX for treatment of breast cancer or lymphoma. We measured the thickening of the media as well as the adventitia of the coronary microcirculatory arteriolar segments.This study was performed at the Wake Forest University School of Medicine as part of a protocol approved by the Institutional Animal Care and Use Committee (Assurance #A04-137) of Wake Forest University, and was funded by the National Institutes of Health (study identifier R21CA109224). A total of thirty-two male Sprague-Dawley rats , gross examination of the rats was performed. The hearts were excised, the aorta was cannulated, and then the heart was mounted on a Langendorff apparatus, pressurized to 52 cm H2O, and perfused with paraformaldehyde for 20 minutes. The heart was then transferred to chilled paraformaldehyde for 24 hours followed by a solution of 70% ethanol. Appropriate sections were trimmed to obtain the middle short axis view. As shown in After animal euthanasia   =  LC/πAverage Lumen Radius (LR)  =  LD/2Medial calculations:Average Medial Diameter (MD)  =  MC/πAverage Medial Radius (MR)  =  MD/2Average Medial Thickness (MT)  =  MR - LRAdventitial calculations:Average Adventitial Diameter (AD)  =  AC/πAverage Adventitial Radius (AR)  =  AD/2Average Adventitial Wall Thickness (AT)  =  AR – MRFull Thickness (FT):FT  =  MT+ATLumen diameter (LD), medial wall thickness, adventitial thickness, and full thickness  were calculated and presented in the absence  and presp<0.05 was considered significant. For all analyses, SAS version 9.1 was used.Animals within each group were assessed at pre-specified time intervals. All analyses that compared groups at a fixed point in time were performed first using analysis of variance (ANOVA), then using analysis of covariance (ANCOVA) to adjust for age, weight, and the total cumulative dose of DOX. In addition, separate analyses were performed to account for the potential influence of time or duration of treatment on our results. Only when there were overall significant differences in appearance among the 3 groups would pair-wise comparisons of least means squares be tested with Student's t-tests between the saline and the low- and high-DOX groups. Comparisons were made to determine whether the thickness of the adventitia, thickness of the media, and the total thickness of the media-adventitia differed between DOX and saline treatment groups. All values are reported as mean ± the standard error of the mean (SEM), and unless stated otherwise, a value of p<0.0001; th week of study , and total thickness (p = 0.02) compared to animals receiving saline  followed by the total wall thickness (p<0.04 for comparisons). In the low DOX and saline groups, the effect of duration of treatment was not significant for any thickness measure throughout the duration of the study.To examine the potential influence of treatment duration on wall thickness values, we categorized duration of exposure into 3 intervals . Because of the relatively few numbers of animals available for study of the 2 week interval, the animals within the 2 and 4 week groups were combined. For the high DOX animals, those with the shortest duration of exposure (2 or 4 weeks) had the highest levels of adventitial, medial and total wall thickness relative to those treated for 7 or 10 weeks. This observation was strongest for the adventitial measures (The results of this study indicate: a) the administration of 2.5 mg/kg/week of DOX increases the thickness of the walls of coronary arterioles of Sprague-Dawley rats, driven primarily by an increase in medial thickening ; b) thesAnthracycline-based chemotherapeutic regimens (including those that incorporate DOX) are among the most widely used regimens to treat malignancies. Today, more than 60,000 patients receive anthracyclines each year to treat leukemia and lymphoma, and breast, uterine, ovarian and lung cancer p<0.001), and stroke  relative to women without cancer or the prior receipt of an anthracycline In addition to CHF, however, anthracycline exposure has been found to increase the incidence of vascular events including MI and stroke As demonstrated in p<0.06 to 0.09) in increases in adventitial and total wall thickness in animals receiving lower doses of DOX after accounting for age, weight and cumulative DOX dose.Since the animals in the study exhibited differences in age, weight, and total cumulative DOX dose, we also performed analyses to account for these differences. As shown in There are several mechanisms which may explain the coronary arteriolar wall thickening observed in this study after DOX. Adventitial hypertrophy has been linked with oxidative inflammation, most notably NADPH oxidase In addition to a primary effect of DOX on the media of the arterioles, other comorbidities may be exacerbated by DOX that indirectly promote increased arteriolar wall thickening. For example, increased medial thickness may result chronically from pressure overload such as elevated blood pressure which may result from increased arterial stiffness The results of this study do not provide mechanisms to account for the increased risk of subsequent MI after anthracycline exposure. Other study results, however, have demonstrated associations between anthracycline exposure and vascular dysfunction in non-coronary vascular beds There are several limitations to this study. First, this may be a phenomenon isolated to rats. As such, subsequent studies will need to be employed in other animal models and human participants to document the occurrence. Second, we are uncertain if this response would occur using other anthracycline preparations. Further studies will need to be performed to see if these changes persist with encapsulated DOX, which is used for human treatment. Third, this study focused on the anatomy of the involved coronary arterioles. Future studies need to also describe the physiologic function of coronary arteries as well as the smaller micro vessels exposed to anthracyclines.In conclusion, early after exposure to anthracycline-based chemotherapy with DOX, microcirculatory coronary arteriolar wall segments develop increased thickening of the medial and adventitial layers leading to an overall increase in microcirculatory arteriolar wall thickness. Further studies are indicated to determine the etiology of this occurrence and whether this finding is permanent and promotes myocardial ischemia in the setting of vasodilator or exercise induced stress.


Enterobacter cloacae BBRC 10061 was used in a method of gradual addition of mazut into medium and its results were compared with simple addition method. To investigate degradation of mazut by BBRC 10061, influence of increase of mazut concentration was assayed based on gradual addition method. Also different kinetic models were used to evaluate kinetics of the process. Results showed that gradual addition method has been a beneficial technique for improvement of mazut degradation because bacterial induction to produce biosurfactant and essential enzymes for cracking mazut was higher during process. Although addition of more mazut increased the rate of biodegradation but percentage of degradation decreased. pH of medium decreased during biodegradation period while electric potential increased. Also the biodegradation kinetics was not fitted with the biokinetic models; therefore kinetics of biodegradation of mazut has to be studied by new models.Mazut as a source content of various hydrocarbons is hard to be degraded and its cracking could turn mazut into useful materials. Nevertheless degradation of mazut by routine methods is too expensive but application of indigenous microorganisms as biocatalysts could be effective and important to lower the costs and expand its consumption. Mazut biodegradation can be improved using various strategies; Therefore in this study newly isolated strain  Pseudomonas aeruginosa, Pseudomonas stutzeri, Provedenica stuartii, Acinetobacter sp, Bacillus Cereus, Micrococcus sp, Enterobacter cloacae and Serratia sp isolated from different locations in Iran which were contaminated by oil compounds, such as soil and water sources, were employed to degrade various hydrocarbons but not mazut [End product of crude oil distillation is a residual compound named mazut that the presence of heavy hydrocarbon compounds such as asphaltene, resins, long chain alkanes, cyclic alkanes and multi cyclic aromatics in it turn mazut into a high viscous and hard degradable product; On the other hand, sulphuric compounds and heavy metals in mazut make use of it more difficult. Mazut has various types based on hydrocarbon compounds which usually depended on geographical area and storage sources which crude oil is prepared from . Bioprocot mazut -15. Biokot mazut ,17. Therot mazut . Each ofot mazut ,20.Enterobacter cloacae BBRC 10061 isolated from oil contaminated soil in Mashhad (Iran), and modulation of its biodegradation kinetics.Objectives of this study were investigation of effect of gradual addition of mazut on biodegradation of hydrocarbon derivative in mazut using an indigenous bacterium E. cloacae BBRC 10061, named NO4 in this study, is indigenous bacteria strain isolated from oil contaminated soil where buses were fixed and fuelled in Mashhad, Iran. It was procured from the biochemical and bioenvironmental research center (BBRC) that was a local culture collection in Sharif University of Technology; Tehran, Iran. The microorganism was maintained in glycerol stock at −20°C for further use.4.7H2O 0.1 g, CaCl2 0.01 g, FeSO4.7H2O 0.01, (NH4)2SO4 5 g, KH2PO4 2.5 g, K2HPO4 2.5 g, sea salt 0.5 g, glucose 4 g and tween80 1 g. The medium was autoclaved at 121°C for 20 min and after cooling, sterile medium was distributed in 50 ml sterile centrifuge tubes which were inoculated with NO4 bacteria amounted to 0.032 OD600.The medium culture had the following composition per liter: MgSOTwo types of culture were used, simple and gradual. To prepare simple culture, 2000 ppm (0.1 g per 50 mL) mazut was added into 50 ml medium culture in centrifuge tube. Gradual culture was prepared at concentrations of 2000 ppm, 10000 ppm and 50000 ppm of mazut. Every 2 day, 400 ppm, 2000 ppm and 10000 ppm mazut were added in centrifuge tubes containing 50 ml medium cultures to prepare gradual cultures in concentrations of 2000 ppm, 10000 ppm and 50000 ppm.All cultivations were then incubated for 10 days at 33°C and initial pH 6.8 with a speed of 160 rpm.For estimation of mazut concentration, spectrophotometer (S2000 UV/VIS) was used. To extract mazut from medium, chloroform was added into medium proportionately with initial concentration of mazut  and centrifuge tube was shaken and blended till all mazut was dissolved in chloroform. Then organic phase content of mazut and chloroform was under and mineral phase was up. To extract organic phase, mineral part voided of mazut and another organic components dissolved in chloroform was decanted. After that, organic phase was sampled and diluted by chloroform solvent. Now absorbance of mazut in medium was measured at 450 nm by spectrophotometer. Sample concentration was measured based on absorbance-concentration curve.600).To estimate bacteria amount in medium, after sampling from medium bacterial density was measured at 600 nm by spectrophotometer and recorded as optical density .s is rate of mazut degradation (mg/ L /day), rs max is maximum of mazut degradation rate (mg/ L/ day), S is mazut concentration (ppm), x is microbial optical density (dimensionless), Ks is half-saturation constant of mazut (mg/ L), Ki is inhibition constant (mg/ L), K1 is inhibition constant (mg/ L), K2 is inhibition constant (mg/ L), λ is constant of Moser model (dimensionless), β is constant of Contois model (mg/ L), A (mg/ L /day), B (day), C (mg /L/ day) and D (mg/ L /day) are constants of linear models. Experimental data for the degraded mazut and the bacteria were used as the experimental data to validate the models. The mathematical models were solved numerically in Microsoft Office Excel 2007 using the SOLVER tool. The kinetic parameters and constants for each timepoint were calculated based on the mazut concentration. A linear regression between the experimental data and the predicted data were computed for all the assays.Results of degraded mazut in this study were fitted on various biokinetic models which are presented in Table Degradation commenced after adding 2000 ppm mazut into medium and during the process NO4 bacteria degraded mazut about 1400 ppm Figure . Biodegr600 (Figure 600. In first days of incubation, pH reduced from 6.8 to 6 relating to increase in microbial growth which made medium acidy (Figure For assessment of effectiveness of gradual method, influence of increase in mazut concentration on biodegradation rate was studied by this method. With increasing additional concentration of about 2000 ppm in which during ten days 10000 ppm was in medium, mazut degradation percentage attained to %30. At this time residual mazut got 7000 ppm and biomass with less fluctuated growth gained about 3 OD0 Figure . Gradual0 Figure . Maximumy Figure , F. ElecBiodegradation of mazut at different concentrations using simple and gradual medium was fitted on the biokinetic models which their results are presented in Table E. cloacae in this study was capable to biodegrade mazut efficiently and was of first bacterial strain cracking mazut. More researches on mazut biodegradation have already been carried out in soil medium whereas this study was completely investigated in controlled specific water condition. Bioremediation of mazut-polluted soil by fungi species reduced 300 mg of initial concentration which was 5000 mg per kg soil during ten days and further incubation up to140 days reduced mazut concentration down to 1500 mg per kg soil [Trichoderma harzianum had highest ability for degradation of mazut [There are just a few studies on biodegradation of mazut compared with other materials such as crude oil and other petroleum compounds. Therefore this project has been a new research and investigation in the field of petroleum biotechnology. New isolated strain of bacterium  kg soil . Among fof mazut . Resultsof mazut . Using gChanges in pH of medium were appropriate for bacterial growth and activity. Simultaneously with higher rate of microbial growth, reduction in value of pH was faster and medium became more acidified. This phenomenon acted as an inhibitor factor for bacterial growth and caused stopping growth. After fast growth of bacteria at starting days, no growth was observed when pH unchanged and became constant. In this study, most amount of reduction of pH was about 1 unit in all of the assays. During the biodegradation of mazut, materials in medium were chemically changed as conductivity of medium was affected by it. Biodegradation of mazut resulted increase of medium’s electric potential that was maximum at growth time. Usually in degradation of hydrocarbons, increase of microbial activities can cause quick reduction of pH which is a reason for reduction of microbial growth; as at following process pH was constant -27. ElecE. cloacae at 50000 ppm, 10000 ppm and 2500 ppm concentrations were %10, %40 and %53 respectively. At high concentrations of heavy crude oil degradation percentage did not change and increasing concentration decreased microbial growth and degradation percentage [Increase of mazut concentration induced bacteria to consume more mazut. Results showed that addition of mazut into medium decreased percentage of biodegradation but more amount of mazut was degraded. Addition of every concentration of mazut could increase degradation but only up to certain amount in which increase in concentration would not raise degradation capacity. After 21 days degradation of heavy crude oil by rcentage . Usuallyrcentage ,28-31; nrcentage ,25 or inIn general, comparing with other studies we could able more clearly show the effective mazut degradation by gradual additional method because there is not any report published about kinetics of biodegradation of mazut. This study could be first in this type of investigation and more studies can be carried out in future. The biokinetic models could be most powerful models, although were not so much correlated with biodegradation of mazut; hence, there is demand for new more powerful model to predict mazut biodegradation process.Mazut were efficiently degraded by newly isolated bacteria. Mazut biodegradation was improved with gradually addition of substrate. Substrate addition allowed bacteria to gradually adapt to a heavy hydrocarbon medium. Gradually adaptation raised biodegradation rate, increasing biomass. Increasing mazut concentration reduced degradation percentage because bacterial growth and consequently, degrading enzymes production were limited in a batch medium. Changing in pH and electric potential of medium were fluctuated during biodegradation. In the early day, decrease of pH and increase of electric potential were occurred; then, their oscillations were reduced. Electric potential change could be evidence for pH alternation, which can help to tune pH in the medium without measuring pH. Investigation of mathematical models on mazut biodegradation stated complexity of mazut biodegradation to mathematically modeling. Therefore, modeling mazut degradation, and using substrate addition technique can be studied to improve other biodegradation processes.The authors declare that they have no competing interests.ACK, carried out this study as part of his M.Sc. project, planed the original research of the project, and wrote the manuscript. MM, supervised the microbial part of this research, and revised the manuscript. SY, supervised the mathematical part of the project, and read the manuscript. All authors read and approved the final manuscript.


The incidence and severity of allergies caused by cedar pollen, house dust, egg protein, and many others are increasing and have recently become a serious social problem. We have previously developed an original reaction . In thisin vitro IgE-inducing system was established using lymphocytes and plasma from donors which were not naturally immunized with allergens. The original system contained ERDF supplemented with fetal bovine serum  and contained human plasma (10%) as an essential component. Human peripheral blood lymphocytes and plasma were obtained by density-gradient centrifugation at 400 × g for 30 min with cell separation medium, Ficoll-Paque™ Plus. This system also included allergen (100 ng/ml), interleukins (IL-) 2, 4, and 6 (10 ng/ml each) and muramyl dipeptide , as described previously [6 cells/ml in the medium and incubated in a CO2 incubator at 37°C for 10 days. During the 10 days, IgE was specifically secreted into the medium.The original The necessity for inclusions of human plasma and interleukins was shown, when human lymphocytes and plasma from donors which were not naturally immunized with allergens were used. For the induction of IgE, human lymphocytes and plasma obtained from the same donor were required . AdditioIn this study, the lymphocytes and plasma of donors naturally immunized with various allergens were used. Therefore, the IgE level of the control was high, i.e., more than 300 ng/ml, as shown in Table In summary, elimination of IL-2 from the IgE-inducing system medium increased the IgE induction level when human lymphocytes and plasma obtained from donors naturally immunized with allergens were used. The level of about 1 μg/ml IgE reported to be secreted in this study may be the highest compared with those reported elsewhere. The original and improved systems for human IgE production are considered to be of profound use for studying allergy mechanisms and surveying allergy-alleviating products, respectively.


The trend of closely related taxa to retain similar environmental preferences mediated by inherited traits suggests that several patterns observed at the community scale originate from longer evolutionary processes. While the effects of phylogenetic relatedness have been previously studied within a single genus or family, lineage-specific effects on the ecological processes governing community assembly have rarely been studied for entire communities or flora. Here, we measured how community phylogenetic structure varies across a wide elevation gradient for plant lineages represented by 35 families, using a co-occurrence index and net relatedness index (NRI). We propose a framework that analyses each lineage separately and reveals the trend of ecological assembly at tree nodes. We found prevailing phylogenetic clustering for more ancient nodes and overdispersion in more recent tree nodes. Closely related species may thus rapidly evolve new environmental tolerances to radiate into distinct communities, while older lineages likely retain inherent environmental tolerances to occupy communities in similar environments, either through efficient dispersal mechanisms or the exclusion of older lineages with more divergent environmental tolerances. Our study illustrates the importance of disentangling the patterns of community assembly among lineages to better interpret the ecological role of traits. It also sheds light on studies reporting absence of phylogenetic signal, and opens new perspectives on the analysis of niche and trait conservatism across lineages. Two main ecological processes are widely recognized to govern the assembly of communities from a regional species pool. First, neutral processes, which encompass demographic stochasticity coupled with dispersal limitations and community drift, create random patterns of species coexistence Hubbell . Second,Indeed, the recent, large interest in investigating phylogenetic niche conservatism . The elevation of the area ranges from 375 m to 3200 m above sea level. Mean annual temperature and precipitation vary, respectively, from 8°C and 1200 mm at 600 m to −5°C and 2600 mm at 3000 m  was amplified by polymerase chain reaction (PCR) using standard forward and reverse primers for angiosperm species , and 1.0 μL of aqueous dilution of DNA. PCR amplification was carried out on an Applied Biosystems GeneAmp 2700 thermal cycler  using an initial denaturation of 2 min at 94°C followed by 34 cycles of 60 sec at 94°C, 60 sec at 50°C of annealing time, 2 min at 72°C of extension, and a final extension of 7 min at 72°C. Resultant PCR products were run on 1.2% agarose gels and stained with ethidium bromide before viewing in GeneSnap . They were purified using a Qiaquick PCR purification kit (Qiagen) before the sequencing reaction. Cycle sequencing of the purified PCR products was performed with the forward and reverse primers in both directions, and an additional primer 20R (5′-TGCATTGC [A/G] CGGTG [A/G] ATGTG-3′) was designed to capture most of the internal part of the rbcL gene. Reactions were performed on ice in 10 μL volumes, each contained 5.0 μL of sterile water, 2.0 μL of sequence terminator ABI Big Dye version 3.1 , 1.0 μL of each primer, and 2.0 μL of cleaned PCR product. Sequencing reaction was carried out on an initial denaturation of 3 min at 96°C, 30 cycles of 15 sec at 96°C, 15 sec at 50°C, and 90 sec at 60°C. Cycle sequencing products were visualized on an ABI 3100 DNA sequencer (Applied Biosystems). Sequences were first checked for identity by BLAST search for highly similar sequences using the NCBI online blast facility. Forward and reverse contigs were edited and assembled with DNA Baser version 3.x  before exported for alignment. We augmented the data to 231 species sequences by downloading 73 rbcL and 123 matK sequences from published sources available in GenBank . CTAB protocols Doyle  were use GenBank . All seqrbcL and matK sequences within each plant family was performed with Clustal W algorithm in Mega  H. Karst were included as outgroup species. The best model of DNA substitution was tested using jModelTest , that no overall trend will be detected between co-occurrence and phylogenetic distances, we assessed the global phylogenetic structure of the regional pool by matrix correlations between pairwise phylogenetic distances and species co-occurrences, and assessed the significance using Spearman's correlations and 9999 randomizations. In order to account for taxonomic and spatial scale in this analysis, we measured phylogenetic structure within two separate groups consisting of monocots and eudicots, as well as in each vegetation zone  along elevation.To test our first hypothesis  is the number of times species S1 and S2 co-occur, while Min  is the occurrence frequency of the rarer of the two species , that communities are more clustered at older nodes, we computed the NRI for each of the 693 communities using the package Picante , that a higher decrease of phylogenetic patterns among angiosperm lineages occurs with elevation, we assessed the relationship between NRI and elevation for each phylogenetic tree node using linear regressions. The t-statistic of the slope coefficient of the regression analyses was then projected on each node of the phylogenetic tree. The significance of this relationship was assessed directly from the regression summary. Linear regressions were used because they provide a simple metric and contain the data output of interest for example estimates of coefficients and 95% prediction interval bands. In addition, the uniformity in sampling efforts across our study area rendered this analysis appropriate for our questions.To test our third hypothesis  Scop. and Cirsium oleraceum (L.) Scop. within the Asteraceae family, and some nodes within the Cyperaceae and Lamiaceae family.The phylogenetic reconstructions, including divergence time estimations, produced well-supported phylogenetic trees with nodes congruent to taxonomic groups defined by the APG III Group  classifir = −0.0014, P = 0.47; Fig. 1) that considering different angiosperm lineages together could result in the absence of a detectable community structure. Similar results were obtained within the monocot  and eudicot  lineages , montane , subalpine , and alpine  along elevation , lineage-specific community assembly revealed both patterns of phylogenetic clustering and overdispersion, as well as no observable trend for various lineages  and 220 , whereas the two most overdispersed nodes were nodes 23  and 228 , the relationship between NRI and elevation showed no strict pattern when considering all angiosperms together  and 154 , whereas the two nodes with the most significant increase with elevation were nodes 198  and 199  can help unravel the ecological structure of communities at fine taxonomic scales  are relatively resistant to low temperatures , we found prevailing patterns of phylogenetic overdispersion in the Apiaceae and Lamiaceae, indicating that closely related species diversified into occupy different communities in contrasting environmental conditions, or close relatives co-occur less often than expected. The observed assembly patterns of these groups may in part be attributed to their life-history traits. For instance, Apiaceae species are characterized by heavy diaspores Körner , which lThe strength of the observed patterns at phylogenetic nodes may depend on the age of species divergence in the various angiosperm families Fig. . Nodes tQuercus species in Florida  and at low elevation . For most plant lineages, we did observe a trend toward decreasing NRI with elevation Fig. , but it Despite the advantage of a multiscale analysis of variation in communities, the lineage-specific approach has some important limitations that deserve mention. First, phylogenetic relationships may not reflect the ecological differences among groups of species that diverged under entirely different ecological conditions Losos . This maThe node-by-node examination of phylogenetic patterns across mountain plant communities proved more informative than a treewide global analysis. Above all, the detailed analysis of community assembly patterns at phylogenetic nodes revealed a rather weak relationship between the phylogenetic relatedness and ecological similarity of species at several nodes. However, it did show that older phylogenetic lineages tended to be clustered in distinct communities under broad environmental conditions, while more recent nodes may have retained some level of ecological diversification into contrasted conditions.Detecting these trends required a lineage-specific approach (Wiens and Graham


Previous studies have shown that cysteine-rich secretory protein containing LCCL domain 2 (CRISPLD2) is a novel lipopolysaccharide (LPS)-binding protein, and the upregulation of CRISPLD2 expression protects mice against LPS-induced lethality. The aim of this study was to examine the expression of CRISPLD2 in patients with sepsis and characterize the association of this protein with procalcitonin.The expression of CRISPLD2 was determined in100 healthy volunteers and 119 septic patients. According to the definition of sepsis, patients were divided into three groups sepsis, severe sepsis, and septic shock. The relationship between CRISPLD2 levels and procalcitonin was also examined and statistically analyzed.The CRISPLD2 levels in healthy individuals were 219.3±69.1 µg/ml. Patients with sepsis exhibited higher CRISPLD2 levels than observed in healthy individuals (p = 0.001), but CRISPLD2 expression was not upregulated in patients with septic shock. No significant differences were observed between the levels of CRISPLD2 in surviving and non-surviving spesis patients. CRISPLD2 levels were negatively correlated with procalcitonin levels.The present study is the first to demonstrate the decreased expression of CRISPLD2 in septic shock and its association with PCT in sepsis. Further studies are needed to clarify the potential association between CRISPLD2 expression and clinical outcomes to determine if it could be used as a novel sepsis biomarker. Sepsis is a major cause of death for critically ill patients in China, the USA and many European countries Our previous work showed that the cysteine-rich secretory protein containing LCCL domain 2 (CRISPLD2/Crispld2) also functions as a LPS-binding protein with significant LPS-binding affinity Thus, the objective of this study was to examine whether CRISPLD2 serum levels are associated with PCT concentration in septic patients and disease severity. If this is correct, then CRISPLD2 in serum levels could be used as a biomarker to predict the clinical outcome of septic patients.This study involved the prospective observation of a cohort of patients admitted to the emergency and intensive care units (15 ICU beds) of an urban university teaching hospital . The ethics committee of Changzheng Hospital in Shanghai, China approved this study. Written informed consent was obtained from the patients or a designated surrogate prior to enrollment in the study.Trained research assistants screened the patients using the hospital’s electronic medical record system. All patients who had been diagnosed with sepsis, severe sepsis, and septic shock were considered for enrollment. Sepsis, severe sepsis, and septic shock were defined according to internationally accepted criteria Healthy individuals from the Health examination center affiliated with Changzheng hospital in Shanghai were enrolled and used in the control group, with an age criteria of >18 years and the patient must not have any acute medical or surgical conditions, that could potentially require admission to a hospital facility. The individuals in the control group who had been diagnosed with or showed evidence of any significant comorbid conditions were automatically excluded.Pertinent patient demographics including age, gender, comorbidities, site of infection, microbial isolates and major laboratory test results were recorded at baseline. All of the participants’ vital signs were measured and recorded at the time of enrollment. The disease severity in each patient was assessed at admission using two different scores: the acute physiology and chronic health evaluation (APACHE) II score We collected blood serum samples within 24 hours after admission. The serum was transferred to plastic tubes and stored at −80°C until further use. The procalcitonin levels were measured using an immunoluminometric assay . The chemiluminescence was measured using a luminometer . CRISPLD2 in serum was measured by competitive ELISA as described in a previous study Data analysis was conducted using SPSS 18 software. The data are presented as the means±SD, median and interquartile ranges  or numbers and percentages. The categorical variables were compared using Chi-square or Fisher’s exact tests, where appropriate.We calculated the differences in parametric data between different strata, using Student’s t test and ANOVA with post hoc LSD test for the two groups. For comparisons of the nonparametric data, the Mann-Whitney test was used for the two groups, and the Kruskal-Wallis test with post hoc Mann-Whitney test was performed for multiple comparisons. Spearman’s rank correlation or Pearson’s tests were performed to evaluate the association between the two groups, where appropriate. We analyzed the data using the receiver operating characteristic (ROC) curve for serum CRISPLD2, PCT levels at admission, and for the 28-day mortality prediction. In all analyses, p<0.05 was considered significant.Patients admitted to the ICU were screened for this study. A total of 380 patients were screened, and 160 did not meet inclusion criteria, and were excluded (without appropriate samples (n = 32) and declining to participate (n = 69)). Subsequently, 119 patients were analyzed in the present study.versus (vs) 46.0 (17.3) years, p = 0.001). Males represented 76.4% of the cohort of patients and 58% ofthe healthy volunteers (p = 0.001). The primary sources for sepsis infection were pneumonia, and abdominal infections, with a predominance of gram-negative bacterial infections. The mortality rate was 26.1% for all patients with a higher mortality for severe sepsis and septic shock.A summary of the patient demographics and clinical parameters of the study population arelisted in The scores generated using APACHEII, SOFA, CCI, the platlet counts, and the creatine and PCT levels differed significantly among the three groups of sepsis patients. The white blood cell count, HCT percent, and total serum bilirubin levels were not significantly different.12/L) were also determined. The CRISPLD2 level was not significantly associated with PCT, WBC, age or sex in healthy population.In the 100 healthy controls, the average CRISPLD2 level was 219.3±61.0 µg/ml; the minimum value was 55.9 µg/mland the maximum was 437.2 µg/ml. The distribution of serum CRISPLD2 is presented in At admission, the PCT concentration differed significantly among four groups: healthy, sepsis, severe sepsis, and septic shock. The CRISPLD2 level was significantly higher in sepsis patients compared with that in the healthy population (p = 0.019) and significantly lower in patients with septic shock compared with that in healthy, sepsis, and severe sepsis . The CRISPLD2 concentration was not significantly different between severe sepsis and healthy (p = 0.161), and no significant difference was observed between severe sepsis and sepsis patients (p = 0.445). .The age, sex, mechanical ventilation, and comorbidity (indicated by CCI) had no effect on PCT and CRISPLD2 concentrations in patients. Patients with abdominal infections had higher PCT and lower CRISPLD2 levels than patients with respiratory infections. Notably, more patients experienced severe sepsis and septic shock through abdominal infections. The PCT concentration was higher in patients with gram-negative isolates than in other patients. The CRISPLD2 level was slightly lower in patients with gram-negative isolates, but there was no statistical significance. .There was a significant negative correlation between the CRISPLD2 serum concentration and the PCT level , SOFA scThe PCT concentration was significantly higher in non-survivors than in survivors, but the CRISPLD2 serum level did not significantly differ between the two groups . The areTo our knowledge, this is the first study concerning the levels of CRISPLD2 in human subjects with sepsis. Here, we conducted a prospective observational study, and found a considerable fluctuation in the CRISPLD2 concentrations of healthy populations compared with sepsis patients. Patients with mild sepsis had slightly higher levels of CRISPLD2 than healthyindividuals. Patients with septic shock exhibited lower levels of CRISPLD2 than healthy, sepsis, and severe sepsis patients. Furthermore, we also observed a statistically significant inverse relationship between serum CRISPLD2 and PCT levels, but serum CRISPLD2 levels between survivors and non-survivors did not significantly differ.CRISPLD2 plays a role in the development of rat lung and mouse kidney and is also involved in the development of nonsyndromic cleft lip with or without cleft palate The data obtained in the present study showed that patients infected with gram-negative bacterial isolates showed a lower level of CRISPLD2 than those with gram-positive bacterial infections. Indeed, CRISPLD2 is not only a LPS-binding protein, but also a single-stranded nucleic acids (ssDNA)-binding protein. Recently, the RAAIH (R3H)-domain, which binds ssDNA, was identified in many mammalian cysteine-rich secretory proteins The results obtained in the present study are consistent with those of previous work Interestingly, low CRISPLD2 concentrations were observed in septic shock. The lower level of CRISPLD2 expression during septic shock might indicate increased utilization or reduced production of this protein. However, the observational nature of the present study makes it difficult to determine which factor plays a major role in sepsis development. Patients with septic shock could consume more CRISPLD2 as a result of persistent bacterial antigen stimulation. A limited ability to produce CRISPLD2 might also contribute to the low protein level observed during septic shock. Many studies have shown that immunosuppressed patients exhibit severe sepsis and septic shock The results obtained in the present study did not show significant differences in the CRISPLD2 levels between survivors and non-survivors. A previous study PCT is a reliable marker to diagnosis and differentiate sepsis, severe sepsis, and septic shock. Within populations of patients affected with sepsis, an elevation of the PCT levels has been associated with poor outcomes in vivo is different compared with LBP. LBP is constitutively expressed in human plasma at low concentrations (3–15 mg/ml) and can increases up to 200 mg/ml during the acute phase response LBP is a major LPS-binding protein in the mammalian biological system, which facilitates the transfer of LPS to the membrane-bound CD14 receptor and then induces inflammation at local sites of infection Glucocorticoids trigger the upregulation of CRISPLD2 expression, as glucocorticoid receptor-binding elements are present in the promoter region of CRISPLD2 Conclusions obtained from research presented here are restricted by the observational nature of this study and limited knowledge of CRISPLD2. First, the current study was not designed to determine which factors might directly or indirectly affect CRISPLD2 concentration in patients. Second, the dynamic observation of CRISPLD2 levels in patients might offer more informations concerning severity and mortality than inferred by the simple determination of CRISPLD2 levels detected on admission in the present study. Third, we did not determine the serum levels of other endotoxin-binding proteins, endotoxins, or other inflammatory markers. Fourth, the LUMI test (Chemiluminescence) was used in this study to examine the PCT levels, which has a lower sensitivity compared with the current assays The decreased expression of CRISPLD2 in septic shock and its association with PCT suggests CRISPLD2 as a potential biomarker in sepsis. Within the limitations of the study, the CRISPLD2 expression was not correlated with the outcome of patients. More research is needed to explore the function of CRISPLD2, the factors affecting the expression of this protein, and its clinical relevance.