Understanding the relative contribution of abiotic and biotic factors to the formation of ecosystem functioning across scales is vital to evaluate ecosystem services. Here, we elucidate the effects of abiotic site conditions  and plant functional traits on variations of stand aboveground carbon (AGC) stock in an old-growth tropical montane rain forest. The response-effect framework in functional ecology is adopted in examining how plant functional traits respond to environmental changes and affect ecosystem functioning. We measured specific leaf area and wood density of 270 woody plant species and estimated stand AGC stocks in a 30-ha forest plot. The relationships among environmental factors (ENVIRONMENT), community-weighted means of functional traits (TRAITS) and stand AGC stocks across nested spatial scales were disentangled by structural equation modeling. The results showed that the stands composed of ‘acquisitive’ species (high specific leaf area and low wood density) had low AGC, whereas stands composed of ‘conservative’ species (low specific leaf area and high wood density) had high AGC. TRAITS responded to ENVIRONMENT and affected AGC directly. ENVIRONMENT had an indirect effect on AGC through its direct effect on TRAITS. TRAITS were more important than ENVIRONMENT in driving variations of AGC. The effects of TRAITS on AGC increased, while the effects of ENVIRONMENT on AGC decreased with the increase of spatial scales in the tropical montane rain forest. Our study suggests that plant functional traits are the mediators in regulating effects of abiotic site conditions on ecosystem functions. A variety of global changes are driving an increase in rates of species extinctions. Biodiversity loss in the 21st century has been one of the major drivers of ecosystem change . Over 60Functional traits are defined as morpho-physio-phenological traits that impact the fitness of individual species via their effects on growth, reproduction and survival, the three components of individual performance . MoreoveThere is a growing consensus that community functional composition determines ecosystem functioning and related services . There aGlobal climatic change as a result of rising level of carbon dioxide in the atmosphere has greatly stimulated the interest in biological carbon sequestration in natural ecosystems, such as forests. Connecting with a series of ecosystem processes, forest carbon stock is considered as one of the paramount indicators of forest ecosystem functioning. An effective climate control by terrestrial ecosystems not only depends on the rate of carbon uptake by primary producers, but also on the rate of carbon release from the biota . Carbon Recent ecosystem research tried to incorporate functional traits into a response-effect framework, which examines the response of plants to environmental change by the “response traits” and plant effect on ecosystem functions by the “effect traits” . StructuMost of the studies have just focused on the relationships between environmental conditions and plant functional traits or on the relations between functional traits and ecosystem functions. A few studies have integrated environment, traits and ecosystem functions into one system to explore the interactions among these three aspects . For an ′–18°50′N, 108°36′–109°05′E) on Hainan Island in southern China. The Jianfengling Nature Reserve is located at the northern edge of the Asian tropical forest, covering ca 640 km2, with an elevation range of ca 0–1,412 m a.s.l. The mean annual temperature is 19.7°C and annual precipitation is 2,651 mm, with a distinct wet season from May to October and a dry season from November to April of the following year. The soil type is latosol, which is broadly distributed in tropical regions of south China (This study was conducted in a 30-ha (500 m × 600 m) forest plot of old-growth  tropical montane rainforest in Jianfengling Nature Reserve  of the randomly selected additional points were located at 2 m away from their associated base points, one third (1/3) were located at 5 m away, and one third (1/3) were located at 15 m away. A soil core with 4 cm diameter and 10 cm depth was collected in each sampling point in 2012 and a total of 585 soil samples were sampled in this 30-ha plot. Soil organic matter (SOM), pH, total and available nitrogen, phosphorus, and potassium, exchangeable potassium, exchangeable sodium, exchangeable calcium, exchangeable magnesium, and exchangeable base (EB) concentrations were determined according to Forest Soil Analysis Methods .2 g-1) was calculated for each lamina as the ratio of leaf surface area to leaf mass. Wood density was determined from increment cores and trunks. The cores were taken from bark to pith of trees (DBH ≥ 5 cm) with an increment borer. Core lengths were then measured with a Mitutoyo digital micrometer and core volumes computed for cylinders of the measured length and inner diameter of the borer. For some shrub species, we had to sample trunk for individuals (DBH ≤ 5 cm) nearby the plot and removed the phloem and bark, measured fresh volume by water displacement. Each sample was quickly wrapped in a sealed plastic bag after sampling. These samples were drying in an oven at 103°C for over 72 h until they became constant weight. Wood density  was computed as the ratio of oven dry mass to fresh volume and averaged for each species.A total of 270 tree and shrub species (belonging to 145 genera and 71 families) was found in the 30-ha plot. The leaf traits were measured for all these species in at least 10 well-growth individuals per species  and wood density was measured for at least five individuals per species  in the plot or nearby the plot from July to October (growing season) in 2011, using standardized protocols for plant functional trait measurements . For eacWe partitioned the 30-ha plot into three spatial scales: 40 m × 40 m (180 subplots), 60 m × 60 m (80 subplots), 80 m × 80 m (42 subplots) and measured the abundance weighted community mean trait for specific leaf area (CWM_SLA) and wood density (CWM_WD) in each scale. CWM trait values were calculated as the mean across species of their trait value weighted by the species relative abundance  using th2H, where ρ is the wood density for each species (g cm-3), DBH is the diameter at breast height (1.30 m in cm) and H is tree height (m). We converted plant woody biomass to carbon stock multiplying it by a factor of 0.5, since carbon stock roughly represents 50% of dry woody biomass . The direct effects were showed in the final SEMs, meanwhile the indirect effects occur if two variables are connected through paths to and from a third variable.To meet model assumptions, some predictors  were a log-transformed. To assess how community weighted means of functional traits affect AGC, we examined the relationships between CWM_SLA/CWM_WD and AGC in each spatial scale by linear regression. Based on the response-and-effect framework . However, wood density at community level (CWM_WD) was positively associated with AGC and the effect of CWM_WD on AGC increased with increasing spatial scale (R2 from 0.28 to 0.36).Results of linear regressions Figure  indicateχ2 = 14.859, df = 8, n = 180, P = 0.062, CMIN/df = 1.857, CFI = 0.986, RMSEA = 0.069), at 60 m × 60 m  and at 80 m × 80 m , respectively. Comparison with the hypothesized model, the non-significant pathway (P > 0.05), the direct effect of ENVIRONMENT on AGC (Environment → AGC) was removed. These final models explained 78%, 67%, and 59% variation of TRAITS for at the 40 m × 40 m, 60 m × 60 m, and 80 m × 80 m scales, respectively, and the explained variations of AGC increased from 29% through 33% to 47%, respectively  among ENVIRONMENT at each scale Figures –D. The dOverall, TC played the most important role in affecting TRAITS and AGC among environmental variables, meanwhile SOM and EB became more and more important on driving variations of TRAITS and AGC with increasing spatial scale. ENVIRONMENT always had a strong effect on TRAITS, though this effect decreased gradually with the increase of spatial scale. TRAITS also had a large effect on AGC and its effect increased with increasing spatial scale. Therefore, TRAITS seemed to act as the mediators in regulating the effects of environmental change on AGC. Moreover, TRAITS always play a larger role in affecting AGC than environmental factors at each spatial scale.Environmental factors can be considered as filters constraining which individuals bearing specific attributes of ‘response traits’ are able to be sorted and persist in a community . DiffereThe response of ‘conservative’ strategy to high SOM was in line with the report of Cation uptake and supply may explain the positive effect of EB on functional traits. Atmospheric inputs of Ca, K, and Mg are usually less than plant uptake, which is typically supplied through mineral weathering, mineralization, and leaching from plant biomass . In our In our SEM, ENVIRONMENT was composed of TC, SOM, and EB. TRAITS composing of high specific leaf area and low wood density were corresponded to ‘acquisitive’ plants with fast nutrient acquisition and turnover. Thus, the negative relation between ENVIRONMENT and TRAITS implies large TC, high SOM and low EB will make community tend to ‘conservative’ strategy (low specific leaf area and high wood density). The tight correlation between variation in habitat preference and plant economics spectrum suggests that the topographically and edaphically associated environmental filter mediated through morphological and physiological traits of species is an important mechanism in shaping spatial patterns of tropical forest community .Carbon storage in vegetation and soil underpins climate regulation through carbon sequestration. Given continuous plant growth, functional traits that optimize acquisition and efficient use of solar radiation and nutrients throughout the forest complex canopy are very important for sustained ecosystem carbon input . In our The role of functional traits at community level has been shown for aboveground net primary productivity  and carbFocusing on plant functional traits rather than on species identities, allows generalizing complex community dynamics and predicting effects of ongoing environmental changes . The fraWe found that although environmental factors always had a strong effect on community functional traits, the effect decreased gradually with the increase of scale size. This could be due to the homogenizing effect of topographic variables at the larger scales (see Supplementary Table We found that the effect of community functional traits on AGC increased with increasing scale size. On one hand, the sampling unit of 40 m × 40 m may be too small to well reflect the local woody species pool of a site in the species rich tropical rainforest. Thus, random effects introduce noise and impede the detection of ecological patterns, which depend on the spatial resolution . On the Functional traits have a potential to describe how the functional composition of communities responds to environmental gradients and how they influence ecosystem processes and services delivery. Furthermore, the relative effects of environmental changes on ecosystem functioning in a rapidly changing world still needs to be evaluated . To diseRZ, YL, and YD designed the study. ML, JW, and CZ collected the field samples and performed the plant traits and carbon stock measurements. WB, HX, and JH analyzed the data and performed the statistical analyses. WB and JH wrote the first version of the manuscript, which was intensively discussed and revised by all authors.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Silica–epoxy nanocomposite models were established to investigate the influence of silane coupling agent on the structure and thermomechanical properties of the nanocomposites through molecular dynamics simulation. Results revealed that incorporating silica nanoparticles into a polymer matrix could improve thermomechanical properties of the composites and increase their glass transition temperature and thermal conductivity. Their thermomechanical properties were further enhanced through silane coupling agent modification on the surface of fillers. Compared with that of pure epoxy, the glass transition temperatures of the silica–epoxy composites with grafting ratios of 5% and 10% increased by 17 and 28 K, respectively. The thermal conductivities of the two models at room temperature respectively increased by 60.0% and 67.1%. At higher temperature 450 K, thermal conductivity of the nanocomposite model with a high grafting ratio of 10% demonstrated a considerable increase of approximately 50% over the pure epoxy resin (EP) model. The elastic and shear modulus of the nanocomposite models decreased at temperatures below their glass transition temperatures. These observations were further addressed in the interpretation from three aspects: segmental mobility capability, radial distribution function, and free volume fraction. Our computational results are largely consistent with existing experimental data, and our simulation model got fully validated. Epoxy resin (EP) has been widely used in power transmission and transformation equipment because of its numerous advantages, such as good adhesiveness, chemical stability, good mechanical properties, flexible processing, low shrinkage, and low cost. With the construction of high-voltage power grids, electrical equipment has been developed in terms of environmentally friendliness, high efficiency, and smaller occupied area. Hence, new dielectric materials with an excellent insulation performance, good aging resistance and adequate mechanical properties have been proposed . HoweverMathematic models and evaluation methodology have been established to assess the comprehensive properties of nanocomposites for practical applications. Electrical, thermal, and mechanical properties can be enhanced through incorporating nanoparticles into polymer hosts, and their effects have been elucidated on the basis of multiple perspectives, such as particle size, shape, amount, and interface interaction characteristics; among these parameters, size and interface interaction possibly play a predominant role ,13,14,15Tg), coefficient of thermal expansion (CTE), elastic modulus, and Poisson’s ratio as a function of different crosslinking degrees, is calculated through molecular dynamics simulations, and computational results are largely consistent with existing principle models [Tg and CTE and obtained results that match well with those from a finite element method [Tg decreases with temperature, whereas the mechanical properties increase with fullerene doping amount [Epoxy resin nanocomposites have been widely simulated through molecular dynamics. A series of thermal and mechanical parameters, such as volumetric shrinkage, glass transition temperature , and free volume fraction calculation, were adopted to explain the predicted data. Finally, simulation models were then validated.Silica, as a common filler of epoxy-hosted dielectric composites, has been rarely examined through computational approaches, although experimental studies have indicated that silica can highly enhance the thermomechanical properties of composites ,31. Sili2/EP), (3) silica–epoxy composite unit cell with 5% grafting ratio of silane coupling agent (SiO2–5%/EP); and (4) silica–epoxy composite unit cell with 10% grafting ratio of silane coupling agent (SiO2–10%/EP). Bisphenol A epoxy resin (DGEBA) was selected as a polymer host, and an aliphatic amine curing agent named 593 is composed of adducts of diethylene triamine and butyl glycidyl ether. The average diameter of the spherical silica particles was 10 nm. The following molecular modeling was conducted:3.Cell construction: single molecular models of DGEBA and the curing agent were manually designed a, and th3, and a time step of 1 fs should be used. In an NPT (constant pressure and temperature) ensemble, Andersen and Berendsen were chosen to regulate temperature and pressure, respectively. The last equilibrated unit cell was utilized for the subsequent crosslinking step.For pre-equilibration, a slow stress relaxation procedure was performed to obtain an equilibrated structure. The geometric optimization of 5000 iterations was conducted to acquire a slightly deformed unit cell with a lower total energy than the initial value. Molecular dynamics simulations were subsequently conducted in an NVT (constant volume and temperature) ensemble for 100 ps at 298 K. In the NVT ensemble, forcefield COMPASS was selected and Andersen was used to control temperature. Afterwards, the unit cell was equilibrated twice under the NPT ensemble at 100 ps and 1 atm to obtain the final density of above 1 g/cm2 group were manually prepared. Then, 16 DGEBA molecules and eight curing agent molecules with active reaction groups formed periodic unit cells and underwent a series of new bond formation processes under the following controlled parameters: initial cutoff distance of 3.5 Å, maximum cutoff distance of 7.0 Å, and conversion degree of 85%. After a specific crosslink PERL language was run, the final model was obtained were used for the molecular dynamics simulations performed in this study. Four kinds of periodic unit cells were prepared to compare the surface of silica particles before and after they were modified using a silane coupling agent. The four unit cells were described as follows: (1) pure epoxy unit cell, (2) silica–epoxy composite unit cell  and temperature. The inflection point can be obtained through the linear fitting of specific volume as a function of temperature, below and above the approximate Tg range, and the corresponding temperature of the intersection point is Tg of the epoxy crosslinking system [V0 and P are the initial volume and pressure of the system, respectively. The linear fitting curve of the specific volume to temperature indicates that the slopes of two straight lines below and above Tg directly represent CTE. The specific volumes of polymers and polymer-based composites change rapidly when their temperature increase as they pass through the glass transition region. Thus, polymeric materials have two different CTEs, one for the glass state (250–400 K) and the other for the rubbery state (425–650 K).g system . CTE refThe established molecular models were used as the initial configuration of our system. After the equilibration at 298 K, the temperature of each unit cell of four kinds of models was increased to 650 K, and the cells were equilibrated again at that temperature at 1 atm for 200 ps. Then, a gradual cooling-down simulation was implemented at a constant cooling rate of 25 K/250 ps until the temperature of each unit cell reached 250 K. The details are as follows:The models were initially equilibrated by an isothermal and isochoric (NVT ensemble) molecular dynamics simulation at 50 ps and 600 K and then by an isothermal and isobaric (NPT ensemble) molecular dynamics simulation at 200 ps and atmospheric pressure. These models were subsequently cooled at a rate of 25 K/250 ps from 650 to 250 K in nine stages, that is, 50 ps—long NVT simulations were performed for each model and 200 ps long NPT simulations were carried out at all temperatures. Thus, an NPT simulation was additionally applied to smaller systems for 200 ps after the cooling-down simulation finished. Finally, the specific volume—temperature plots were obtained after the curve was linearly fitted.To investigate the effects of grafting modification of particles on the thermal and mechanical properties, the diameter of nanoparticles was fixed 10 Å and volume fraction was 7.2 vol % for nanocomposites models. Side length of unit cell for pure epoxy resin model was about 30 Å and slightly increased with temperature, whereas, the side lengths of cubic unit cells for the other three nanocomposites models varied in the range of 39 to 41 Å, reflecting the stability of energy. Correspondingly, densities of four models have good agreement with computational and experimental results ,37.Thermal conductivity is an important parameter to represent the thermophysical property of EP materials. This parameter should be improved to enhance the heat dissipation capability of EP materials in order to maintain a relatively lower operating temperature of equipment.V is the total volume of the system, kB is the Boltzmann constant, T is the temperature unit, and J is the microscopic heat flux of the system. However, this method is characterized by several disadvantages, such as slow computation speed and poor convergence. As such, an innovative NEMD method has been introduced to solve thermal conductivity through Fourier’s law under the condition of a fixed temperature difference and heat flux.Thermal conductivity has been investigated in relation to molecules or atoms. On the basis of the analog computation of molecular dynamics, we can summarize thermal conductivity investigations as equilibrium molecular dynamics (EMD) and nonequilibrium molecular dynamics (NEMD) methods referring to equilibrium and nonequilibrium states, respectively . AccordiRNEMD aims to determine heat flux by exchanging the velocity vector of atoms inside a system. In this study, the thermal conductivity of EP composite models could be simulated through this method because the total energy and momentum of the system remained constant.xy, xz, and yz are respectively subjected to small shear deformation along the x-, y-, and z-axis directions to produce 12 groups of strains with the extent of strain maintained within 0.01 in accordance with Hooke’s law, as shown in Equation (3):Mechanical properties can be determined by calculating small deformations at different static pressures in given directions. The planes of E), shear modulus (G), bulk modulus (K), and Poisson’s ratio (υ), of the EP composite system can be expressed as follows:Within the error range, EP crosslinking can be simulated and calculated as an isotropic material. Therefore, stiffness matrix is simplified as follows:Tg was marked as the corresponding temperature at the inflection point of the slopes of specific volume–temperature linear curves. Tg and the calculated CTE of the four models are summarized in Achieving a complete equilibrium state is difficult because of a large number of atoms in a molecular system. Therefore, molecular dynamics calculation can be applied to reach a balanced state when the density of a particular system fluctuates in a small range around the target temperature. In general, 200 ps is set to obtain a well-equilibrated system after NPT simulation is performed. Tg was evident when SiO2 particles were added to the epoxy host, especially when Tg of SiO2–epoxy model with 10% grafting ratio of silane coupling agent increased by 7% compared with the pure epoxy resin model. Tg of SiO2/EP composite matched well with experimental data reporting Tg of 141.2 °C (414.35 K) in Ref. [Tg also increased possibly because of the high conversion degree. By contrast, CTE declined below and above Tg of the silica–epoxy composite model. Surface modification with the silane coupling agent could obviously reduce CTE and could directly demonstrate the formation of a tight 3D cross-linked structure with a small freedom space in the system.An increasing tendency of 2 °C 414.5 K in ReTg on the cooling rate as shown in Equation (9) [Tg values from simulation and experiment (ΔTg) to the relative cooling rate:To further interpret the existing difference among the computational results, the Willliams–Landel–Ferry (WLF) equation can be used to quantitatively account for the dependence of tion (9) . The WLFTg between experimental measurements  and experimental value range of (363–420 K) [In Equation (9), 6.77 K/s ) and our3–420 K) .The common mechanical and electrical properties of epoxy resin must be maintained during long-term operations. However, the temperature of devices with epoxy resin materials inevitably increases. As such, a feasible design should be proposed to produce dielectrics with enhanced thermal properties. Usually, the temperature of epoxy resin materials must be kept below 450 K during long-term operations. Therefore, the thermal conductivity of EP composites between 250 and 450 K were investigated through molecular dynamic simulation. In 2 is 7.2%. Maxwell’s equation (Equation (10)) revealed a calculated thermal conductivity of 0.261 W/(m·K), which is slightly smaller than the simulated result (0.284 W/(m·K)) based on the molecular dynamics method, but this error is also within the allowable range:For the silica–epoxy composite model without grafting, the volume fraction of SiO2 nanoparticles can improve the thermal conductivity of epoxy resin. As temperature increases, without grafting procedure, thermal conductivity increases from 33.04% at 300 K to 44.75% at 450 K compared with that of the pure EP model.The four models show a linear increase in thermal conductivity from 300 to 450 K. Incorporation of SiO2 nanoparticle surface can considerably enhance the thermal conductivity of composites. A high grafting ratio corresponds to a considerable increase in the thermal conductivity of composite models. At 300 K, the thermal conductivity of SiO2–10%/EP model increases by 67.07%, which is higher than that of the pure EP model.Grafting silane coupling agents on SiOWe can obtain the following conclusions for the molecular dynamic simulation of thermal conductivity:14, C15, C16, C24, C25, C26, C34, C35, C36, C45, C46, and C56, whereas the silica–epoxy models are isotropic in terms of the similar values of C11, C22, C33, C12, C13, C23, C44, C55, and C66. Lamé constants can be determined on the basis of the presumed isotropic characteristics of the silica–epoxy model in Equations (11) and (12):The mechanical properties of the four models at different temperatures were simulated in the Forcite Package of Materials Studio. On the basis of the calculated stiffness matrix, we observed that the pure EP model exhibits anisotropic characteristics in terms of nearly zero values of CTg is improved after SiO2 is incorporated. The calculated results are consistent with those reported in previous studies [2 nanoparticles with silane coupling agents can improve the mechanical properties of epoxy resin.According to Equations (4)–(8), the elastic and shear modulus of the four models as a function of temperature are shown in  studies ,31,32. TEtotal represents total energy of the composite models, EEP is the total energy of the pure EP model, and ESiO2 is the total energy of SiO2 nanoparticles. The combination between polymer hosts and nanoparticles forms new chemical or hydrogen bonds that play important roles in establishing a 3D cross-linked structure. The binding energy of composite models can be calculated using Equation (13) to elucidate the connection mechanism after grafting procedure:In 2 particles can increase the interaction energy between SiO2 and EP and thus enhance the mechanical properties of silica–epoxy composites [Therefore, grafting silane coupling agent on the surface of SiOmposites ,31.2/EP, SiO2–5%/EP, and SiO2–10%/EP) were analyzed on the basis of the kinematic capacity of the model molecular chain to determine the effects of temperature on the thermodynamic properties of these models. The total simulation time was 50 ps, and one frame was the output for every 0.5 ps. The MSD curves of the four models at different temperatures are shown in The MSDs of the four models increase with time until a steady state is reached. At a high temperature, especially above 450 K, the value increases faster, which can be attributed to the resultant larger kinetic energy of the molecular chains when the models attain the glassy state. A strong kinetic energy of a molecular chain can lead to a rapid decline of mechanical properties. These results are consistent with the calculated mechanical properties.2 nanoparticles without surface modification into the epoxy matrix may increase the MSD compared with that of the pure epoxy resin model. However, the silica–epoxy model with 10% grafting ratio of the silane coupling agent can reduce the MSD shown in Incorporation of SiOThe plots of the mean square displacement (MSD) as a function of time at different temperatures in the four models , and this value was higher than that of the pure epoxy resin model at 0.214 W/(m·K). The binding energy between SiO2 and epoxy resin also increased by up to 38%, demonstrating a strong interaction between polymer hosts and particles after grafting.Incorporating SiO2 particles increased. The model of SiO2–10%/EP also shows a lower value of free volume fraction than that of the model of SiO2/EP. Overall, grafting procedure proves necessary to control and maintain the improved mechanical and thermal properties.The analysis of mean square displacement, radial distribution function, and free volume fraction further elucidates the improved mechanism. Temperature drastically influences the three values. The silica–epoxy model with 10% grafting ratio of silane coupling agent could create a tight 3D cross-linked structure and maintain a relatively low MSD value at high temperatures. In the radial distribution function analysis, the peak representing the hydroxyl group at 0.96 Å became stronger than that of the pure epoxy resin model, verifying that the amount of –OH on the modified surface of SiOIn this study, the mechanical and thermal properties of silica–epoxy composite models were investigated through molecular dynamics simulation. Our results confirmed that surface modification by grafting a silane coupling agent onto a nanoparticle surface is an efficient method to improve the mechanical and thermal properties of composites. The following findings can be obtained through simulation calculation:


Phaseolus vulgaris).Rhizobia are alpha-proteobacteria commonly found in soil and root nodules of legumes. It was recently reported that nitrogen-fixing rhizobia also inhabit legume seeds. In this study, we examined whole-genome sequences of seven strains of rhizobia isolated from seeds of common bean ; this represents the first GTA reported for this genus.Rhizobial strains included in this study belonged to three different species, including Seeds represent a niche for bacteria; their access by rhizobia possibly triggered the infection of phages, recombination, loss or gain of plasmids, and loss of symbiosis genes. This process probably represents ongoing evolution that will eventually convert these strains into obligate endophytes.The online version of this article (10.1186/s12864-018-5023-0) contains supplementary material, which is available to authorized users. Triticum aestivum [Zea mays [Oryza sativa [Arabidopsis thaliana [Populus euphratica [Rhizobia belong to the group of alpha-proteobacteria and thrive in diverse environments, either as free-living bacteria in soil or as symbionts within the root nodules of legumes, where they fix atmospheric nitrogen . Rhizobiaestivum , Zea mayZea mays , Oryza sa sativa , Arabidothaliana , and Popphratica .Phaseolus vulgaris) [R. phaseoli, R. leguminosarum, R. grahamii, and Sinorhizobium americanum. Some of these strains form pairs with other strains based on the plasmid profiles; however, only one of the members comprising the pair is capable of fixing nitrogen [Recently, we showed that nitrogen-fixing rhizobia also occur within the seeds of common bean (ulgaris) ; this exnitrogen . We perfnitrogen .Agrobacterium pusense isolated from root nodules of un-inoculated bean plants; the source of these bacteria is thought to be seeds [Agrobacterium strains are closely related to the strain IRBG74, which is currently the only Agrobacterium isolate with a symbiotic plasmid, capable of forming nitrogen-fixing root nodules in Sesbania cannabina [High similarity has also been shown among strains of be seeds . These Aannabina , 12. Notannabina .It is important to determine the genomic sequences of these rhizobial strains and to identify the sequence variation resulting from the loss of symbiotic ability in some of them, as this information would help elucidate the process of evolution and functional consequences of changes in gene content. In this study, we report the genomic sequences of seven rhizobial strains isolated from the interior of bean seeds. We analyzed these sequences to understand the modifications in chromosomes, plasmids, orthologous gene sequences, and gene functions that possibly emerged as a consequence of the arrival of these strains in the seeds.R. phaseoli , R. grahamii (CCGM3), and related to R. leguminosarum . The R. phaseoli strain CCGM1 was isolated in the same experiments with the rest of strains and was included in this study for comparison with CCGM2. Both strains form a highly related pair [We analyzed the genomes of seven rhizobial strains isolated from nodules of un-inoculated common bean plants. Characteristics of these strains and of those used for comparisons are summarized in Table rpoB  in the case of CCGM3 and CCGM6. The sequences retrieved from PCR products were 100% identical to the corresponding segment in the sequenced strains. The strains CCGM4 and CCGM8 did not present symbiotic plasmids; and thus, plants inoculated with them will not yield seeds. Strain CCGM5 was not detected in seeds. Strain CCGM9 was not included in the greenhouse assay.Of the strains analyzed here, four were detected by PCR in seeds harvested from plants inoculated with these strains; these included CCGM1, CCGM2, CCGM3, and CCGM6  was present only in CCGM9 . Both these strains shared the same plasmid profile, with the exception of a 240 kb plasmid, which was present only in CCGM4  , with the exception of some segments that were absent from all seed strains  and pCCGE502b (1.3 Mb). However, the seed strain CCGM3 contained three plasmids, two of which were similar to pCCGE502a and pCCGE502b, whereas the third plasmid appeared to share homology with a 1 Mb chromosomal segment of CCGE502  were orthologous among the strains  showed ANI genomic values of 92% in comparison to the nodule strain; this value was below the threshold level (95%) accepted for assigning a strain to a given species.To assess the overall sequence homology among strains analyzed here, we calculated the average nucleotide identity (ANI) and genome coverage  in seed strains belonging to different groups. For example, transcription (class K), intracellular trafficking (U), carbohydrate metabolism (G), amino acid metabolism (E), and synthesis of cofactors (H) were enriched in R. phaseoli; intracellular trafficking (U) and synthesis of cofactors (H) in strains related to R. leguminosarum; and prophages and transposases (X) in R. grahamii. By contrast, the nodule rhizobia were enriched in genes of unassigned function (not in COGs) and in R. grahamii CCGE502 also in defense (V).To identify genes that were potentially important for the survival of seed rhizobia, functions of genes present only in seed strains were compared with those present only in nodule strains . The prophage Pseudo_JBD showed high sequence similarity with one prophage of strains related to R. leguminosarum, CCGM4 and CCGM5, suggesting that this prophage was transferred to bean seeds at the same time as the rhizobial seed strains. The strain CCGM2 shared three prophages with the strain CCGM1, all of which are commonly found in rhizobia and are related to Brucel_Pr. Strain CCGM2 also had four small genomic fragments related to prophages found in Gordonia and Pseudomonas. Genomes of strains CCGM8 and CCGM9 carried five identical prophages, and these were related to some of the prophages found in Synechocystis, Bacillus, Staphylococcus, and Gordonia  was 92% identical to CCGM1 prophage Pseudo_JBD. The Rhodov_vB prophage was inserted at the same chromosomal position, i.e., in the tRNA-Leu gene, in the strains CCGM4, CCGM5 and CCGM1. The other prophage was almost identical between CCGM4 and CCGM5, and was similar to Sinorh_phiM9. The strain CCGM6 harbored very different prophages, with only one related to Rhizobiales (Rhizob_RRA1) and the others related to prophages found in lla Fig. .R. grahamii strain CCGM3 harbored two probable prophages. One of these prophages was predicted to be complete (Paraco_vB_pMaS_IMEP1) and belonged to a new class of prophages, referred to as gene transfer agents (GTAs) [The genome of s (GTAs) , 15. MorGenomic sequences of seven new seed rhizobial strains were obtained and analyzed in this study. In addition to the genomes of four strains analyzed previously , 10, seqThe bioinformatics approach used here was very helpful in detecting the overall relatedness between seed and nodule strains Fig. . These aAgrobacterium strains (CCGM10 and CCGM11) has been published previously [A. pusense; only one strain of this species, IRBG74, is capable of forming root nodules in legumes to fix nitrogen [Agrobacterium strains, such as fixK or fixNOQP.Of the strains examined in this study, six formed three pairs of highly related strains Fig. . A similnitrogen . StrainsSome of the seed strains are ineffective in forming root nodules and fixing nitrogen; this raises the question as to how these strains accessed the nodules. We propose that effective rhizobial strains or their clones originally formed these nodules, and the ineffective strains co-inhabited the nodules. The pair of rhizobial strains comprising CCGM1 and CCGM2 possibly represents a special case. Although both these strains were effective in nodulation and nitrogen fixation and showed almost identical genomes, they differed by one plasmid.Plants can sanction ineffective rhizobial species or their strains with inferior performance . The symR. leguminosarum, based on their 16S rRNA or rpoB sequences [R. leguminosarum, R. l. viciae, R. l. phaseoli, and R. etli strains. The highest ANI value obtained for these strains was 92%. In a strict taxonomic classification, strains CCGM4, CCGM5, and CCGM6 should be renamed as Rhizobium sp.The determination of ANI was essential for estimating the sequence similarity between strains and for identifying highly related strains  and nitrogen fixation  are arranged in clusters in the symbiotic plasmid, whereas other genes, such as the symbiotic cytochrome oxidase Cbb3 and transcriptional regulator fixL are reiterated in other plasmids [Plasmids are important for bacteria to monitor and respond to environmental fluctuations . Rhizobiplasmids . Our funObligate symbionts undergo drastic genome reduction because some of their housekeeping functions are fulfilled by the host or because some defense mechanisms are no longer needed –30. On tAbout half of the genomes in bacterial databases contains genes of prophages , and theR. grahamii strain CCGE502, 1 Mb of chromosomal DNA was homologous with a plasmid of CCGM3 , infection of phages, loss of plasmids, and rearrangement of replicons. The overall outcome of this process was the successful establishment of strains in this new niche. However, we detected ineffective sister strains due to the loss of genomic segments harboring genes necessary for nodulation and nitrogen fixation. Further research on similar microorganisms is needed to understand their dynamics with the plant and to explore their utility as seeds carrying their own inoculum.2).Rhizobial strains were isolated from root nodules of un-inoculated plants as described previously . Strains540) of 0.1. Plants were maintained in the greenhouse at a temperature ranging from 22 °C to 29 °C, 60% humidity, and natural photoperiod. Plants were irrigated alternately with sterile water and Fahraeus medium [Seeds of common bean cultivar Negro Jamapa were surface sterilized with 20% sodium hypochloride for 5 min and rinsed extensively with sterile water. Seeds were then immersed in sterile water for 1 min and rinsed several times. For germination, seeds were arranged in a tray lined with moist paper and covered with aluminum foil. The tray was incubated in the dark for 3 days at 29 °C. Seedlings were transferred to sterile pots filled with vermiculite, and each plant was inoculated with 1 ml of freshly prepared bacterial suspension with an optical density at 540 nm . PCR products were sequenced at the Sequencing Unit, Institute of Biotechnology-UNAM, Cuernavaca, México. Reaction conditions and primers used for PCRs are listed in Additional file The high molecular weight plasmids were visualized using the Eckhardt technique, as modified by Hynes and McGregor ., et al., 2015).Genomic DNA was extracted from the rhizobial strains, following standard protocols , and useR. phaseoli CIAT652, R. leguminosarum WSM2304, and R. grahamii CCGE502. The genome sequence of CCGM1 strain, which we previously characterized [R. leguminosarum strain WSM1689  was also included in this analysis. Graph of prophages was generated using Circos [Pairwise comparisons were made using ProteinOrtho v5.15  between cterized , was inccterized . The geng Circos .The genomic sequences of each strain were deposited in the GenBank database under the provisional accession numbers shown in Table Statistical differences were tested with the Fisher’s exact test.Additional file 1:Detection of rhizobial strains in bean seeds via PCR amplifications. (PPTX 148 kb)Additional file 2:List of genes present in strain CCGM5 and absent in CCGM4. (XLSX 189 kb)Additional file 3:Average nucleotide identity (ANIm) and genome coverage of seed and nodule rhizobial strains. (PPTX 181 kb)Additional file 4:Comparison of gene functions covered by seeds strains compared with nodule strains. (PPTX 206 kb)Additional file 5:Shared and strain-specific genes in the analyzed rhizobial strains. (XLSX 455 kb)Additional file 6:Genes with repeated function (by name). (XLSX 10 kb)Additional file 7:Complementary data on the detection of prophages in the genomes of rhizobial strains. (XLSX 25 kb)Additional file 8:Sequence of primers and PCR conditions. (DOCX 14 kb)


Symptomatic epidural hematoma (SEH) after anterior cervical spine surgery is very rare, but it has disastrous consequences for the patients. Timely diagnosis and evaluation can effectively reduce the sequelae of neurological deficit in SEH. The purpose of this study was to retrospectively analyze a subset of clinical data of SEH after anterior cervical spine surgery, and to investigate the risk factors and treatment experience of this serious complication.Neurological deterioration after anterior cervical spine surgery was detected in six patients. Epidural hematoma was confirmed by emergency cervical magnetic resonance imaging (MRI). The patients included five males and one female, with an average age of 56.7 ± 13.1 years (range 42–76 years). Three patients had a history of drinking and/or smoking. All of the patients were treated with nonsteroidal anti-inflammatory drugs (NSAIDs) preoperatively, but without anticoagulant drugs or pre-spinal surgery. The coagulation function was normal in all patients. Except for one patient, who had lower blood pressure (BP) during the operation and higher BP after the operation, the other patients had a normal level of BP during the pre-, intra-, and post-operation periods. The average time was 9.9 ± 6.7 hours  from the postoperative period to the initial neurological deficit and 6.3 ± 6.0 hours  from the initial deterioration to evacuation. Five patients underwent emergency evacuation, and one patient underwent conservative treatment. Four patients who underwent evacuation and one patient who received conservative treatment achieved neurological function recovery with an American Spinal Injury Association (ASIA) grade 2.4 ± 0.9  score at the last follow-up. One patient with confirmed arterial epidural hemorrhage during the evaluation showed no neurological function recovery at the last follow-up.Wide exposure of the epidural space and BP level during the perioperative period play an important role in the formation of SEH after anterior cervical spine surgery. Arterial epidural hematoma has serious consequences; therefore, early diagnosis and evaluation play an important role in the recovery from paralysis. SEH after spinal surgery is rare, and its reported incidence is 0.1% to 0.41% in the literature.,6–8 However, the complications are very serious and often lead to a neurological deficit, especially for cervical surgery, which can lead to paralysis and even death.,11Since Jackson–14 An analysis of the results shows some limitations,–11 such as low incidence and few cases. This study reviewed six cases with clinical data of SEH following cervical diseases  and cervical spondylotic myelopathy (CSM)) in the past 10 years, and it aimed to provide more possibilities for the etiology of SEH, more options for the treatment of such complications, and more options for the prevention of such diseases.In a few studies the presence of SEH after anterior cervical surgery was reported, and the influencing factors and treatment strategies were analyzed.2All medical history data collected in this article were obtained from the patients and confirmed by them. This study was approved by the Ethics Committee of the Affiliated Hospital of Southwest Medical University (KY2019147). From January 2009 to December 2018, we treated 6890 patients with CSR and CSM. Neurological deterioration was observed after the operation, and epidural hematomas were confirmed by cervical MRI in 6 (0.09%) cases, including 5 males and 1 female, with an average age of 56.7 ± 13.1 years . We retrospectively analyzed the history, drug treatment, BP during the pre-, intra-, and postoperative periods, hematoma evacuation surgery, complications, and neurological function of these 6 patients. Neurological function was assessed by the ASIA grade.3Table Table 44.1Patient 2, male, 42-year-old, was admitted to the hospital for radiation pain and numbness in the right upper extremity since the last 2 years. After obtaining the medical history, and performing comprehensive physical and imaging examinations, shown in Figure At 8.7 hours after surgery, the patient's sensation disappeared below the level of C6 and perianal and saddle areas, muscle strength was 0 grade, ASIA grade was grade A, tendon reflexes disappeared, and pathological signs were negative. The drainage tube was unobstructed, and it drained 10 ml of bloody fluid. Mannitol and prednisolone were used, and cervical MRI examination was planned immediately. Epidural hematoma was detected at the C5-thoracic 2 (T2) level behind the vertebrae on the MRI image  surgery under general anesthesia. The mean BP level was about 73/55 mmHg, and no bleeding was detected during the operation. After the patient recovered from anesthesia, limb numbness was relieved significantly when compared with the preoperative level. The muscle strength of the limbs was 4/5 and the ASIA grade was grade D. The patient returned safely to the ward. Her BP was 160/88 mmHg post-operation.Two hours after the operation, the patient again complained of numbness and weakness in her limbs. Physical examination showed sensory disturbance of the extremities, key muscle strength of limbs 3/4, and ASIA grade D. Limb tendon reflexes were weak and the pathological signs were negative. The drainage tube was in its normal position and unobstructed, and it drained 20 ml of bloody fluid. Considering the possibility of hysteria, mannitol and prednisolone were given immediately. Seven hours after surgery, paralysis was not relieved but it had aggravated, and ASIA grade decreased to grade A. an emergency cervical MRI showed an epidural hematoma at the C5−C7 spinal canal level , and all patients with epidural hematoma had received more than one disc-segment surgery, which was similar to that suggested by other authors.,15 Extensive anterior or posterior segmental cervical surgery increased the damage to the epidural venous plexus, which may be the main cause of intraspinal venous plexus injury.,13 The wall of the intraspinal venous plexus is weak, and there is no venous valve in the lumen. Bleeding after injury does not stop spontaneously. Henderson showed that the pressure of the intraspinal venous plexus is lower than that of the cerebrospinal fluid. When the pressure of the hematoma, which was caused by intraspinal venous plexus bleeding, approaches or reaches the same pressure as that of the cerebrospinal fluid, it can compress the wall of the intraspinal venous plexus to achieve hemostasis. In our medical records, two patients with anterior cervical single-segment ACDF developed paralysis postoperatively and MRI confirmed the presence of an epidural hematoma. Paralysis of these two single-segment ACDF patients had recovered, one with evaluation and the other without evaluation. The reasons for this occurrence may be that single-segment ACDF surgery causes less trauma, small exposure, and milder intraspinal venous plexus injury. The four patients treated with ACCF all had large exposure of the epidural space and extensive intraspinal venous plexus injury, which were one of the most important reasons for postoperative hematoma. As the pressure exerted by the hematoma increases up to the level of cerebrospinal fluid pressure, the venous plexus hemorrhage stops automatically.Amir believe that compression of the spinal cord is mainly caused by arterial bleeding in the spinal canal. Guodong believes that arterial hematoma in the spinal canal can be relatively low, because arterial bleeding can be easily detected during surgery. According to our medical records, none of our 6 patients had a history of hypertension before surgery. In patient 4, arterial bleeding was found in the epidural space. No active arterial or venous bleeding was found during the initial operation. We speculated and analyzed the following possible causes: the BP was low in the initial intraoperative period (mean 73/55 mmHg); therefore, small arterial hemorrhage in the epidural space was not detected and bipolar coagulation was not adequate. When the patient recovered from anesthesia, BP (160/88 mmHg) increased and the small arteries that do not achieve adequate hemostasis started to bleed again, resulting in the formation of epidural hematoma. Therefore, we conclude that a hematoma caused by epidural arterial hemorrhage will lead to severe cord injury, and the recovery from paralysis after evacuation will also be very poor, and a hematoma caused by epidural venous plexus hemorrhage will also lead to cord injury, but paralysis may recover by at least grade 1 (ASIA score) after the evaluation.Hypertension is an important risk factor for postoperative hematoma, but none of reported studies has elaborated the possible mechanism by which BP causes epidural hematoma. Some authorsThis study is based on retrospective analysis of medical records. There is a lack of adequate evidence and literature reports to confirm the correctness of our speculation, but we still recommend that the BP should be strictly controlled during the intra- and post-operative periods for ACDF or ACCF. During the in-operation period, the systolic blood pressure (SBP) should not be lower than 90 mmHg and the diastolic blood pressure (DBP) should not be below 60 mmHg to accurately observe the presence of active bleeding in the decompression area. Postoperative BP control  also reduced the risk of plexus bleeding.,11,15,19 In this study, average patient age (56.7 ± 13.1 years) and mean blood loss (163.3 ± 117.8 ml) were less than those in the literature reports. All of the six patients were irregularly treated with NSAIDs, but they were not treated with anticoagulation before the operation. Therefore, we do not have sufficient evidence to prove that SEH is related to age, coagulopathy, NSAIDs, anticoagulation, and excessive bleeding. Three patients (50%) in this group had a history of smoking. Two of them had lower levels of smoking, and 1 patient had quit smoking 3 years before the operation. Three patients (50%) had a history of alcohol consumption, and only one patient fulfilled Amir criterion of 10 units of alcohol consumption per week. Although we do not have enough evidence to confirm the relationship between SEH and smoking or drinking, we still need to pay attention to smoking and drinking, which may be one of the risk factors for SEH after cervical surgery in the literature.The other etiologies, such as age more than 60 years, NSAIDs, anticoagulant drugs, excessive bleeding (>1000 ml), drinking (1 week > 10 units), and smoking rate, may be the risk factors for SEH. especially for epidural arterial hemorrhage. Patients who underwent evaluation within 24 hours showed significant recovery from paralysis when compared with the preoperative level, and there was no chance of recovery from paralysis if the evaluation was performed after more than 24 hours.,20,21 In our group, 5 patients underwent evaluation within 12 hours, and 4 patients showed recovery from paralysis of at least two grades at the last follow-up. The results were satisfactory. In patient 4, paralysis lasted up to the last follow-up. The main reason for this occurrence was that hysteria was initially considered when the patient showed neurological symptoms. Cervical MRI examination was not arranged in a timely manner, and all of these methods failed to diagnose the epidural hematoma earlier and accurately. Therefore, we believe that timely cervical MRI is necessary when there is re-deterioration of neurological function or rapid progress of paralysis after anterior cervical surgery. The hematoma caused by epidural arterial hemorrhage develops rapidly and causes serious damage to the spinal cord. Earlier diagnosis and evaluation play an important role in the recovery from paralysis.Timely diagnosis and treatment can significantly increase the chances of recovery from paralysis,www.letpub.com) for its linguistic assistance during the preparation of this manuscript.We thank LetPub (Data curation: Yehui Liao, Yang Tian.Formal analysis: Yehui Liao.Investigation: Chao Tang, Qiang Tang, Rupei Ye.Methodology: Dejun Zhong, Fei Ma.Project administration: Dejun Zhong, Yehui Liao.Resources: Sheng Yang, Hongchun He.Software: Chao Tang.Supervision: Dejun Zhong.Validation: Dejun Zhong, Yehui Liao.Visualization: Sheng Yang, Chao Tang.Writing – original draft: Yehui Liao, Yang Tian.Writing – review & editing: Yehui Liao, Dejun Zhong.


Understanding the drivers of the structure of coral reef fish assemblages is vital for their future conservation. Quantifying the separate roles of natural drivers from the increasing influence of anthropogenic factors, such as fishing and climate change, is a key component of this understanding. It follows that the intrinsic role of historical biogeographical and geomorphological factors must be accounted for when trying to understand the effects of contemporary disturbances such as fishing.Comoros, Madagascar, Mozambique and Tanzania, Western Indian Ocean (WIO).We modeled patterns in the density and biomass of an assemblage of reef‐associated fish species from 11 families, and their association with 16 biophysical variables.a), and reef exposure or slope. Where spatial effects were not significant between mainland continental locations, fishing effects became evident explaining 6% of the variation in data. No correlation with live coral was detected. Only 37 species, predominantly lower trophic level taxa, were significant in explaining differences in assemblages between sites.Canonical analysis of principal coordinates revealed strong country affiliations of reef fish assemblages and distance‐based linear modeling confirmed geographic location and reef geomorphology were the most significant correlates, explaining 32% of the observed variation in fish assemblage structure. Another 6%–8% of variation was explained by productivity gradients (chl_Spatial and geomorphological histories remain a major influence on the structure of reef fish assemblages in the WIO. Reef geomorphology was closely linked to standing biomass, with “ocean‐exposed” fringing reefs supporting high average biomass of ~1,000 kg/ha, while “lagoon‐exposed fringing” reefs and “inner seas patch complex” reefs yielded substantially less at ~500kg/ha. Further, the results indicate the influence of benthic communities on fish assemblages is scale dependent. Such insights will be pivotal for managers seeking to balance long‐term sustainability of artisanal reef fisheries with conservation of coral reef systems. Occasionally, only three replicate transects were collected when only one dive was possible. All individual fish were identified to species level, with the exception of the Balistidae which were categorized as benthic or planktivorous; the Pomacanthidae which were recorded as invertivores (Pomacanthus spp. + Pygoplites diacanthus and Apolemichthys spp.) or grazer‐detritivores (Centropyge spp.)  Table . Trophic.) Table . Small is et al. .2.5The complete fish dataset consisted of density and biomass for 156 species across 53 sites. Preliminary explorations identified two extreme outlier sites , which were therefore removed. To further reduce variance, any species occurring <3 times across all sites were removed. The final dataset for analyses consisted of 45 sites  for 123 coral reef‐associated fish species/taxa  transformed biomass data. One‐way PERMANOVAs confirmed the significance of country differences for both abundance and biomass at (P(Perm) < 0.001). Sites were therefore coded by country a priori for the CAP. We identified which fish species were most influential in describing differences between countries by superimposing vectors representing Pearson correlations (|>0.45|) of individual species with the CAP axes  were run to investigate potential drivers of the observed reef fish assemblage patterns : Centropyge spp. and Pomacanthus spp.; and invertivorous balistids. While the first three species of this group were highly abundant across several locations, densities of the other species varied considerably  and illustrated that sites in Comoros and Madagascar strongly separated from those in the adjacent mainland countries of Mozambique and Tanzania  which were found in most sites throughout Tanzania and Mozambique, and were highly abundant in some of these sites  and biomass data  and the CAP analysis identified clear separation of fish assemblages between countries  being more abundant in Mozambique, whereas large C. sordidus  were more abundant in Comoros  in Shomoni, Grande Comore. This species was highly significant in defining the fish community in all countries based on the SIMPER analysis of density data  were also significant, but at a reduced level. The direction of these variables on the species’ abundance matrix is shown as vectors in the CAP ordination  and reef geomorphology were the most significant, explaining 12.8% and 18.9% of the variation in the dataset, respectively. Reef exposure (4.1%) and chlorophyll_a explained 5.0% of the variation. However, reef slope rather than exposure was significant, explaining 3.3% of the variation in the data. In summary, the CAP analyses and DistLM showed that space (latitude/longitude), reef geomorphology, chlorophyll_a, exposure, and slope were valid predictors of the structure of the fish assemblages across the region. Notably, none of the benthic variables, including live coral cover, were significant.The best‐fit DistLM identified similar clear separation of fish assemblages between countries based on biomass data Figure b, explai3.3At the smaller spatial scale of Tanzania and Mozambique, the DistLM found geomorphology explained the highest variation in the density data at 23.3%, with exposure and fishing pressure also significant explanatory variables, at 6.5% and 6.0%, respectively. The same variables were also significant correlates of the biomass data with geomorphology, exposure, and fishing pressure explaining 22.7%, 6.5%, and 6.5% of the variance, respectively. Thus, at this smaller spatial scale, fishing pressure became a significant variable for both fish density and biomass. However, again, none of the benthic variables were significant.3.4Total fish biomass, based on all 12 families, was low to moderate in all sites in the Comoros at <558 kg/ha and highly variable within the other three countries depending on the location of the survey site Figure . For exa44.1a; reef exposure and reef slope. Since the WIO represents a biogeographic province of similarity in fish species distributions  is not a key factor , and was notably significant for planktivores and piscivores , reef location, isolation, and connectivity Mora, , may helor Mora, , our res4.3Fishing has been shown to precipitate top‐down and bottom‐up trophic cascades on coral reefs , Pomancanthus spp. (invertivores) and Centropyge spp. (grazer‐detritivores), Sufflamen spp. (invertivores), and Acanthurus leucosternon and A. nigrofuscus (grazers), A. tennenti (grazer‐detritivores), Naso elegans (browser), and Ctenochaetus striatus (detritivore). The functional role of these species within the reef ecosystem is likely driven by trophic pathways , and Tekamaji, Mozambique , suggesting these sites may represent reefs with crystalline waters. Some species were only seen on the east African mainland such as the small piscivorous grouper, Cephalopholis argus, and the planktivorous Caesionidae which are an important food source for piscivores (Hobson, a levels on the mainland. The snapper, Lutjanus fulviflamma was widely distributed and highly abundant at some sites Tanzania and Mozambique but not observed in Comoros. It is likely that few or no mangroves near most sites in Comoros may be the reason for the lack of L. fulviflamma since their juvenile phase is almost entirely in mangroves (Kimirei, Nagelkerken, Mgaya, & Huijbers, C. argus at sites in Comoros and Madagascar is not easily explained since both taxa are abundant in the Chagos Archipelago (Samoilys et al., A. dussumieri, A. blochii, and A. xanthopterus and the grouper Plectropomus punctatus. Again, the influence of detritus and algae is suggested by these acanthurids. Plectropomus punctatus is a widespread grouper endemic to the Indian Ocean that is susceptible to fishing pressure, suggesting this may be the reason that it was uncommon or unsighted in Comoros, Tanzania, and Mozambique. The distribution pattern of the size and hence trophic group of the widely distributed excavator, Chlorurus sordidus, may also reflect fishing pressure. Larger individuals were more abundant in Comoros and smaller individuals more abundant in Mozambique, possibly because parrotfishes are not targeted in Comorian artisanal fisheries (Freed & Granek, Other species significant in delineating differences in assemblage structure are highlighted here because they may serve as useful indicators for reef type and health. These included the detritivorous  Hobson, . High deChaetodon meyeri and C. lineolatus, were among the significant species in delineating the differences in assemblages between countries. It is therefore surprising that none of the six benthic variables in our study were significant in explaining patterns in fish assemblages, even at the smaller scale of mainland Africa. In contrast, significant associations between live coral cover, recently dead coral, and rugosity and the structure of fish assemblages were found in the Chagos Archipelago (Samoilys et al., Climate change‐induced coral bleaching is unquestionably one of the primary impacts occurring on coral reefs today (Hughes et al., 5The observed differences in the structure of fish assemblages across the WIO support the growing understanding that comparing reef fish assemblages across large spatial scales has to first take into account reef geomorphology, other reef structural attributes, and larger scale environmental drivers such as nutrient levels (Heenan et al., None declared.MAS conceived the ideas. MAS collected the data. AH led the data analysis. MAS led the writing. AH contributed to the writing. KO contributed ideas, data analysis, and writing. Click here for additional data file. Click here for additional data file. Click here for additional data file. Click here for additional data file. Click here for additional data file. Click here for additional data file.


Many children with neurofibromatosis type 1 (NF1) have focal abnormal signal intensities (FASI) on brain MRI, whose full clinical impact and natural history have not been studied systematically. Our aims are to describe the clinical and neuroradiological features in children with NF1 and cerebellar FASI, and report on the natural history of FASI that display atypical features such as enhancement and mass effect.A retrospective review of the hospital charts and brain MRIs was performed on children from Manitoba diagnosed between 1999 and 2008 with NF1, who also had cerebellar FASI on MRI.Fifty patients  were identified. Mean duration of follow up was 10.1y. Developmental delay, learning disabilities, tumors, and visual signs occurred commonly. Cerebellar signs were not reported. Mean age of the patients at baseline MRI was 7.8 (SD: 4.5) years. FASI occurred in several brain locations and were rarely confined to the cerebellum. FASI displayed mass effect and enhancement infrequently but were associated with malignancy only once. The number of FASI at baseline MRI was significantly less in patients with attention deficient hyperactivity disorder and more if a first degree relative had NF1 or if they had decreased visual acuity.Patients with NF1 and cerebellar FASI do not have motor or consistent non-motor  cerebellar features. The number of FASI may correlate with some clinical features. FASI may display enhancement and mass effect but they rarely become malignant. Neurofibromatosis type 1 (NF1) is an autosomal dominant neurocutaneous disorder with an estimated incidence of 1/3500 live births . This, cPatients with NF1 are at increased risk of developing both benign and malignant tumors compared to those in the general population . The mosPatients with NF1 may have behavioral impairment and cognitive dysfunction. While many studies have been done on the subject, results are often conflicting and no clear NF1 cognitive profile has emerged. Visuospatial and fine motor deficits have long been considered hallmark neurological features of NF1 , 10. MulMany attempts have been made to correlate these cognitive and neurological deficits with radiologic findings. The most common abnormalities found on brain MRI are regions of increased signal visible on T2-weighted images i.e., focal abnormal signal intensities (FASI). These are believed to be present in 60–80% of pediatric patients with NF1 , though The cerebellum plays an important role in the processing of non-motor in addition to motor tasks. Many studies support non-motor roles for the cerebellum in cognition . ChildreThe aims of this study are to: Describe the clinical features in children with NF1 and cerebellar FASI on MRI, correlate the clinical features, especially developmental delay, learning disabilities, and examination findings, with baseline neuroradiological abnormalities, and describe the natural history of FASI that display atypical features including mass effect and enhancement.Electronic MRI neuroimaging reports stored in the radiology information system at our Health Sciences Centre (HSC) for the 10-year period starting January 1, 1999 were read and flagged for the occurrence of posterior fossa abnormalities. Patients with NF1 who had cerebellar radiological abnormalities mentioned on any brain MRI report when they were less than 17 years old between 1999 and 2008, were selected. All patients were seen in Winnipeg Children’s Hospital for a clinical evaluation. Patients from neighboring provinces who attended Winnipeg Children’s Hospital were also included. Ethical approval for the study was given by the Research Ethics Board of the University of Manitoba.Inclusion criteria for the study were: (1) Patients were less than 17 years old during the study period, (2) patients had NF1, and (3) patients had cerebellar abnormalities on MRI. Exclusion criteria were: (1) Patients with solid central nervous system tumors unrelated to NF1, (2) trauma involving the posterior fossa, (3) significant brain malformations and anomalies unrelated to NF1, (4) patients who had brain radiotherapy or chemotherapy before their first brain MRI, and (5) patients with other unrelated brain disease e.g. demyelinating disorders.The hospital chart, pediatric neurology clinic letters, and genetic clinic letters were reviewed in each of these patients. The relevant data available until 2013 were extracted and entered into a purpose-specific database. The data consisted of demographic information, duration of follow-up period, pregnancy, birth and perinatal history, family history of NF1, presenting symptoms, presence of features seen in NF1 including café-au-lait spots, axillary freckling, neurofibromas, bone dysplasia, NF1 related tumors; other diagnoses, developmental milestones, learning disabilities, and physical exam findings especially ocular and central nervous system examination abnormalities.Brain MRI was acquired on 1.5- or 3-Tesla MRI scanner (GE) using standardized protocol with sagittal T1-weighted, axial/ coronal T2-weighted, and axial/ coronal fluid-attenuated inversion recovery (FLAIR) images. Supplemental imaging sequences were performed as needed including T2*, DWI, ADC (apparent diffusion coefficients) maps, fast spoiled gradient echo (FSPGR) images, and MRA. Contrast with Gadolinium was given at the discretion of the radiologist. All initial and subsequent brain MRI images available that were completed by June 2014 were reviewed independently by two pediatric radiologists with expertise in neuroimaging. Disagreements were resolved by consensus.Age at the time of the MRI scan, structural and signal abnormalities in the cerebellar vermis, cerebellar hemispheres, brainstem, and supratentorial structures including the cortex and white matter, basal ganglia, thalami, hypothalamus, and optic nerves/ chiasm were recorded, as well as the presence of cerebellar hypoplasia  or cerebellar atrophy (shrunken size with prominence of the cerebellar folia). Optic nerve(s) and/ or chiasm gliomas (optic pathways gliomas) were diagnosed when these structures were enlarged/ thickened on MRI. Detailed information was collected on FASI on FLAIR images including their number, locations, diameter (defined as a straight line passing from side to side through their center and thus representing their maximum length), signal characteristics, uptake of contrast on T1-weighted images, and mass effect on adjacent structures. Brain locations were divided as follows: cerebellum, brainstem , thalamus/ hypothalamus, basal ganglia/ internal capsule  , and cerThe data were converted to numerical variables using a numerical coding scheme to render the data suitable for statistical analysis and modelling. Mean and median were used to describe the normally distributed and skewed data, respectively.Fifty patients fulfilling the inclusion criteria were identified. Their mean age at the end of the study period was 16.1 years . There were 27 males and 23 females. Mean duration of follow up was 10.1 years. Table N = 50, SD: 4.5 years, median: 7.3 years, minimum: 1.2, maximum: 18.2 years), where SD is the standard deviation. Table A few brain MRI scans during the early part of the study period were not available for review. Fifty baseline (or the first available) brain MRI scans were reviewed. In addition, if any FASI displayed atypical features then all subsequent available MRI scans were reviewed. Mean age of the patients on the first MRI was 7.8 years . The youngest was 14 months old. In those four patients FASI was present in one or more other brain locations  in addition to the cerebellum.A single patient (#54) had widespread numerous FASI in the cortex and subcortical white matter of both cerebral hemispheres. To minimize extreme skewness of the data, we excluded this patient from the analysis involving cerebral FASI counts.N = 49, minimum: 0.8, maximum: 5.6 cm). In one patient the image was degraded by motion artifact hence no measurement was done. FASI with the longest diameter were mostly located in the cerebellum on the first MRI (in 22 of 49 patients). In several instances FASI with the longest diameter, especially those > 2.5 cm, was caused by several confluent FASI that could not each be measured individually and separately.The longest diameter of FASI irrespective of its brain location among our patients in the first scan had a median length of 1.6 cm . On the other hand, there was a trend for the hypothalamus to be involved in the youngest patients (age 0–0.9 year) in comparison with older patients i.e., 1–1.9, 2–4.9, and 5 years or older at their first clinic visit (p = 0.055).There was a significant relationship between the occurrence of FASI in the basal ganglia and age of the patients at symptoms onset. FASI in the basal ganglia occurred at a significantly later age (2 years or older) in comparison with the younger age groups (0–0.9 and 1–1.9 years) at symptom onset (p = 0.033). A similar trend was also found for developmental delay and thalamic FASI (p = 0.08). On the other hand, a trend between developmental delay and the presence of FASI in the hypothalamus was present (p = 0.075). Otherwise, no other relationship between FASI location and other clinical symptoms was seen. In addition, none of the clinical symptoms showed any significant relationship with various combinations of FASI in different brain locations.Learning disabilities were significantly less likely in the presence of FASI in the thalamus (p = 0.009). In addition, abnormal visual fields were significantly less likely with thalamic FASI (p = 0.044), while abnormal visual fields and fundi exam were significantly more likely with cerebral FASI . Multiple logistic regression of cerebral involvement with symptoms and signs showed that the odds of cerebral involvement in children with abnormal fundi was 7.46  times higher than normal fundi after adjusting all other symptoms and signs variables.As for clinical signs, univariate analysis revealed that strabismus was significantly more likely to be associated with basal ganglia FASI (p = 0.079), while abnormal fundi were significantly associated with widespread involvement of FASI in all brain regions (p = 0.009). However, on multivariate analysis where funduscopy and visual fields were paired, the latter association was no longer significant.When various combinations of brain locations of FASI were correlated with clinical signs, there was a trend for abnormal visual fields not to be associated with the presence of FASI in all of the following locations: cerebellum, thalamus or basal ganglia, and brainstem (p = 0.01) and in patients with impaired visual acuity (p = 0.019) after adjusting for all other clinical features. The expected number of FASI at baseline scan for patients with ADHD was 39.3% lower than for those without ADHD after adjusting for all other variables . In addition, the expected number of FASI at baseline scan for patients with impaired visual acuity was 60.1% higher than for those with normal visual acuity after adjusting for all other variables .When the clinical features were correlated with the total number of FASI on baseline MRI, significant associations were found in patients with attention deficient hyperactivity disorder (ADHD) (p = 0.076) for a positive history of NF1 in any family member to be associated with the total number of FASI on baseline MRI. However, this association became significant if a first degree relative rather than a more distant relative had NF1 (p = 0.029). The expected number of FASI at baseline scan for patients with a first degree relative with NF1 was 56.1% higher than for those with no family history of NF1 after adjusting for all other variables.There was only an overall trend .Of the 50 patients, 18 had optic pathways gliomas on their initial MRI scan. Abnormalities in the visual acuity, visual fields, and funduscopy exam were significantly associated with optic pathways gliomas on baseline brain MRI (p = 0.019). Furthermore, the mean number of FASI in the supratentorial region and in the cerebrum (but not in other brain locations) were each significantly higher in patients with optic pathways gliomas (N = 18), than in patients without optic pathways gliomas (N = 31) on baseline MRI .There was a significant association between the presence of optic pathways gliomas on baseline MRI and the number of patients with FASI in the cerebrum but not in other brain locations. Cerebral FASI was present significantly more commonly in patients with optic pathways gliomas, than in patients without optic pathways gliomas on baseline MRI . With every one-year increase in maternal age at conception, the expected number of infratentorial FASI at baseline MRI was 2.75% lower. Similarly, there was a significant relationship between maternal age at conception and the number of brainstem FASI on baseline MRI (p = 0.029). With every one-year increase in maternal age at conception, the expected number of brainstem FASI at baseline MRI was 3.98% lower. There was only a similar trend between maternal age at conception and the number of basal ganglia/ internal capsule FASI on baseline MRI (p = 0.067).There was a significant relationship between maternal age at conception (available in 41 patients) and the number of infratentorial FASI on baseline MRI (p = 0.065) and ADHD (p = 0.081) in children with NF1.Two trends were found for associations between decreasing maternal age at conception and the increased presence of learning disabilities , and similar to other studies  in our cohort in comparison with several studies 4–25%) , 25, 27.% , 25, 2On the other hand, cerebellar motor signs were absent in our cohort, consistent with a normal cerebellar motor function despite the presence of cerebellar FASI. While clumsiness, lower manual dexterity score, and impaired fine motor skills or coordination has been reported in NF1 patients, the occurrence of frank cerebellar ataxia has not , 28, 29,The nature of FASI is unclear but may be related to increased fluid within the myelin associated with hyperplastic or dysplastic glial proliferation as suggested in a study using newer MRI techniques, e.g. multi-exponential T2 relaxation and diffusion MRI including diffusion tensor imaging and diffusion kurtosis imaging . DevelopStudies documenting the neuroimaging-pathological correlation of FASI are rare. In one study, three pediatric patients with NF1 had autopsy which showed that the pathological correlation of FASI were areas of fluid-filled vacuolar or spongiotic change . There wBrain MRI scans in our children with cerebellar FASI showed that FASI rarely occur in isolation and most commonly occur in multiple brain regions in addition to the cerebellum, especially the brainstem and basal ganglia as noted before , 19, 22,There was a suggestion that age at symptom onset or first clinic visit may correlate with where FASI develops, since FASI that developed in the basal ganglia occurred in patients who were relatively older at age of symptom onset. There was a tendency to develop hypothalamic FASI in patients who were relatively younger at the age of their first clinic visit.FASI developed below the age of 2 years in different brain locations in a few of our patients, which is not considered to be typical by some authors , and espPrior studies have shown that cognitive deficits were more common in patients with NF1 and thalamic FASI , 12, espAs for clinical signs, visual fields and especially funduscopic abnormalities were associated more commonly with patients who had cerebral FASI. This can be explained through confounding with the presence of optic pathways gliomas on baseline MRI, since cerebral FASI was seen significantly more commonly in patients whose baseline MRI also showed optic pathways gliomas. Strabismus was significantly associated with the presence of FASI in the basal ganglia. We found no studies that directly link strabismus with disorders of the basal ganglia. In one study of 213 children with cerebral palsy, strabismus was reported in 3 of 15 children with dyskinetic cerebral palsy. Abnormalities in the basal ganglia are implicated in dyskinetic cerebral palsy . Our finThe total number of FASI at baseline MRI was significantly less in patients with ADHD and more if a first degree relative had NF1 and also more in patients with decreased visual acuity. This latter association may have arisen through the occurrence of optic pathways gliomas, which themselves were associated with higher mean numbers of FASI in the supratentorial regions and more specifically in the cerebrum. The former two associations i.e. ADHD and family history of NF1, may be caused by genetic factors/ predisposition but this is speculative. FASI was not associated with the presence of ADHD in 76 NF1 patients, who were preselected and did not have epilepsy or optic nerve glioma .The relationships between decreasing maternal age at conception and the increasing number of FASI at baseline  or the increasing presence of learning disabilities and ADHD  are intriguing; albeit, small in magnitude. They raise the question on whether decreasing maternal age at conception per se or through confounding factors such as socioeconomic status or educational achievement , somehowA few FASI showed contrast enhancement, mass effect, or both. Such findings are reported to occur rarely , 41, 42,Limitations of our study include incomplete ascertainment of patients, missing data, variable durations of follow up, variable ages at which the MRI scans were done, non-uniform MRI magnetic strength and protocols used in these patients, difficulties associated with measuring the diameter of ill-defined FASI, and parental report of learning disabilities were not formally verified through comprehensive cognitive assessment.Patients with NF1 and cerebellar FASI do not have cerebellar motor symptoms or signs. If ataxia develops then neuroimaging is indicated. Patients with NF1 and cerebellar FASI may or may not have learning disabilities or developmental delay and therefore, cerebellar FASI cannot be implicated directly as a causative factor for non-motor cerebellar symptoms. FASI occurred in several brain locations and were rarely confined to the cerebellum in our preselected patients. FASI has the potential, though uncommonly, to show mass effect and/ or contrast enhancement. Rarely, such features herald a malignant change.The total number of FASI at baseline MRI was significantly less in patients with ADHD and more if a first degree relative had NF1 and also more in patients with decreased visual acuity. The latter association is likely caused by the presence of optic pathways gliomas in our cohort. The influence of maternal age of conception on the development of FASI or its association with a few clinical features requires further study.


The present study aimed to assess the effectiveness of clam extract in combination with atorvastatin against experimentally hyperlipidemia in rats. Forty male rats were divided into 5 groups (8 rats /group): control, high fat diet (HFD), atorvastatin (AROR), clam extract (CE), and ATOR + CE. The treatments with ATOR and /or CE significantly reduced the body weight gain, AST, ALT, ALP, TL, TC, TG, LDL-C, urea, creatinine, and uric acid levels while they increased total proteins, albumin, and HDL-C. The treatment with ATOR only did not cause any significant change in CK and MDA along with antioxidant system, while the treatment with CE alone or with ATOR significantly decreased CK and MDA accompanied by improving the antioxidant system. Combination of CE extract with atorvastatin improved the hyperlipidemic efficacy and reduced undesirable side effects especially on muscle. Obesity is one of the highest public health epidemics of the 21st century with about 2 billion adults worldwide currently classified as being overweight or obese [There is significant evidence that elevated oxidative stress mediates the progression of many of the comorbidities associated with obesity . EpidemiStatins consider the standard drug in the treatment of hypercholesterolemia and the management of patients with increased cardiovascular disease risk . StatinsCoelatura aegyptiaca) is a Molluscan Bivalve belonging to Unionidae common in the Egyptian Nile River [ Coelatura aegyptiaca extract in combination with atorvastatin against experimentally hyperlipidemia in rats.Many natural products have hypolipidemic effects and can be used in treatment of obesity as alkaloid extract of the cyanobacterium Spirulina platensis , echinocle River . Many inle River , hepatople River , and chole River , 19. TheAtorvastatin powder and Carboxymethyl cellulose (CMC) were purchased from Sigma-Aldrich . Kits of biochemical parameter were purchased from Biodiagnostic Company .Coelatura aegyptiaca) were collected from the River Nile at Giza Governorate, Egypt. Whole bodies away from the shells were stored at -20°C until needed. After thawing, CE was prepared as follows: whole bodies (1 kg) were coarsely chopped and homogenized with a mixer. The homogenate was extracted in a boiler with 1 liter of distilled water for 3 hrs and allowed to cool to room temperature. After filtration, the filtrate obtained was then concentrated and dried using a lyophilizer , the dose chosen according to Soliman [Freshwater clams ( Soliman .Rattus norvegicus) weighing 140±10 g were used in this study. The rats were obtained from the National Research Center . Rats were housed in a temperature and humidity controlled environment and given food and water ad libitum.Male albino Wistar rats  (Egypt) (CUFS/F/33/15). All the experimental procedures were carried out in accordance with international guidelines for the care and use of laboratory animals.The rats were fed a high fat diet (HFD) with energy of 5.3 kcal/g, comprising 40% calories from fat, 17% from protein, and 43% from carbohydrate for 4 weeks according to a modification of the protocols of Reed et al. . The dieAfter one week of acclimatization, forty rats were divided into five groups (8 rats /group).Control Group. After 4 weeks of normal diets feeding, the rats were given orally 1 ml of 0.5% CMC, daily for 16 days.HFD Group. The rats were fed HFD for 4 weeks and then were given orally 1 ml of 0.5% CMC, daily for 16 days.Atorvastatin Group (ATOR). After 4 weeks of HFD feeding, the rats were given 1 ml of ATOR  daily for 16 days [ 16 days .Clam Extract (CE) Group. After 4 weeks of HFD feeding, the rats were given 1 ml of CE  daily for 16 days [ 16 days .ATOR + CE Group. After 4 weeks of HFD feeding, the rats were given 1 ml of ATOR  and after 1 hour were given 1 ml CE  daily for 16 days.Body weight was measured weekly and body weight change was measured from difference between final body weight and initial body weight. Liver and kidney weighed and their indices were calculated (Tissue  index = (tissue  weight/body  weight) × 100 ).After the end of all experiments, the rats were fully anesthetized with 3% sodium pentobarbital, and the chest was opened. A needle was inserted through the diaphragm and into the heart. Negative pressure was gently applied once the heart had been punctured, and the needle was repositioned as required until blood flowed into the syringe. The blood collected from the rats was separated by centrifugation  to obtain sera which were stored at −80°C for the biochemical measurements.Liver, kidney, and skeletal muscles were removed and were immediately blotted using a filter paper to remove traces of blood. Parts of the tissues were stored at -80°C for biochemical analyses and other parts were suspended in 10% formal saline for fixation preparatory to histopathological examination.Liver, kidney, and skeletal muscles tissues were homogenized (10% w/v) in ice-cold 0.1 M Tris–HCl buffers (pH 7.4). The homogenates were centrifuged at 3000 rpm for 15 min at 4°C and the resultant supernatant was used for the oxidative stress analyses.μm and stained with hematoxylin and eosin (H&E) according to Bancrof and Stevens [The liver, kidney, and skeletal muscles tissues were sectioned at a thickness of 4-5  Stevens , as rout Stevens . The hisThe serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were estimated by the method of Reitman and Frankel ; serum aMDA level is an index of lipid peroxidation and it was estimated by Ohkawa et al. , glutathValues were expressed as means ± SE. The comparisons within groups were evaluated utilizing one-way analysis of variance (ANOVA) with Duncan post hoc test used to compare the group means and p < 0.05 considered statistically significant. SPSS, for Windows (version 15.0), was used for the statistical analysis.The body weight gain, liver weight, kidney weight, and their indices increased significantly (p< 0.05) in HFD group as compared with control group, while the treatments with ATOR and / or CE significantly (p< 0.05) reduced theses parameters as compared to HFD group .Our data shown in Data recorded in The liver of control rats is formed of the classic hepatic lobules. Blood sinusoids were seen separating the cords of the liver cells and lined by flattened endothelial cells and von Kupffer cells . The hisMicroscopic examination of kidney sections of control groups showed normal appearance of the tissue where glomeruli appear as dense tufts of capillaries enclosed in the outer layer of Bowman capsules. Numerous renal tubules were observed . Kidney Microscopic examination of extensor digitorum longus muscle of control group revealed the normal structure of skeletal muscle . Muscle Obesity is the current medical as well social problem, leading to increasing morbidity and mortality. The main risk factors of hyperlipidemia are associated with atherosclerosis, which predisposes ischemic heart disease and cerebrovascular disease . The preOur results revealed that the body weight gains of the rats in the HFD group were significantly higher than that in the control group, and the weight gains in the ATOR, CE, and ATOR+ CE groups were normalized to or lower than that in the HFD group. The consumption of a HFD facilitates the development of a positive energy balance and leads to an increase in visceral fat deposition . HoweverLiver biomarkers enzymes “AST, ALT, ALP” are monitoring the liver structural integrity and damage . In the In the current study the administration of HDF resulted in dyslipidemic changes as illustrated by increasing TL, TG, TC, LDL-C, and low level of HDL-C. Dyslipidemic changes occurring in obesity may be due increased influx of excess nonesterified fatty acids into the liver . While tAbnormalities in lipid metabolism appear to play a pathogenic role in development of renal diseases , 50. In Obesity is related to skeletal muscle loss and dysfunction and the development of muscle atrophy . Also, fOxidative stress is produced through increased reactive oxygen species (ROS) and/or decreased antioxidant mechanisms . High faCombination of CE extract with atorvastatin improved the hyperlipidemic efficacy and reduced undesirable side effects especially on muscle.