The amount of boron waste increases year by year. There is an urgent demand to manage it in order to reduce the environmental impact. In this paper, boron waste was reused as an additive in road base material. Lime and cement were employed to stabilize the waste mixture. Mechanical performances of stabilized mixture were evaluated by experimental methods. A compaction test, an unconfined compressive test, an indirect tensile test, a modulus test, a drying shrinkage test, and a frost resistance test were carried out. Results indicated that mechanical strengths of lime-stabilized boron waste mixture (LSB) satisfy the requirements of road base when lime content is greater than 8%. LSB can only be applied in non-frozen regions as a result of its poor frost resistance. The lime–cement-stabilized mixture can be used in frozen regions when lime and cement contents are 8% and 5%, respectively. Aggregate reduces the drying shrinkage coefficient effectively. Thus, aggregate is suggested for mixture stabilization properly. This work provides a proposal for the management of boron waste. In China, its reserve has reached 35 million tons, and emission still grows at a rate of 1.6 million tons per year [Boron waste is discharged in the production of borax pentahydrate, borax decahydrate, anhydrous borax, per year . Its envIt can be seen from et al. [et al. [et al. [Ozdemir et al.  added bo [et al.  studied  [et al.  investiget al. [et al. [et al. [et al. [et al. [For the purposes of sustainable development and industrial ecology, boron waste has been gradually used in the concrete and brick. Topçu et al.  investig [et al.  prepared [et al.  also poi [et al.  demonstr [et al.  utilized [et al. .et al. [It can be seen that reuse of boron waste in brick and concrete increases the managing efficiency. For concrete, its content should be less than 15%. For brick and lightweight aggregate, its amount should be less than 30%. Remarkably, the strength of concrete and brick obviously decreases when the content exceeds the limited value. This means that boron waste cannot be massively applied in high strength material. Therefore, further research is needed to explore other reuse methods. Fortunately, Modarres et al.  proposedet al. [et al. [et al. [In order to find a good stabilized method, many studies have been performed in recent years. Qian et al.  investig [et al.  used cem [et al.  studied Based on the above analysis, we can find that boron waste has a significant impact on the environment. Its reuse is helpful for protecting the environment. In order to increase the reused amount as much as possible, it was reused in the road base material in this study. Lime and cement were used to stabilize this waste. The stabilized waste mixture was prepared in a laboratory. Specimens were then made according to the standards JTG E51-2009 , and theBoron waste, which was discharged in the production of borax, was used in this study. Its color is light brown, as shown in uC) and coefficient of curvature (cC) are also given in Basic properties of boron waste are given in The content of effective calcium oxide and magnesium oxide in hydrated lime is 60.4%. Ordinary Portland cement with a level of 42.5 MPa was used in this study. The properties of the cement are listed in 3. The properties of the soil were tested according to the Test Methods of Materials Stabilized with Inorganic Binders for Highway Engineering of China (JTG E51-2009) [Limestone aggregate and soil were selected for experiments. The apparent density of aggregate was 2.677 g/cm51-2009) , as giveHydrated lime and cement have been widely used to stabilize soil and granular material ,17,18. TUnconfined compressive strength (UCS) is an important index of geo-materials ,21,22. AP is the ultimate load, N; H is the height of specimen, mm; and d is the diameter of specimen, mm. a is the width of loading strip, mm. α is the center angle corresponding to half width of the strip, rad. E is resilient modulus, MPa.An indirect tensile strength (ITS) can be used to evaluate the anti-cracking performance of geo-materials . Therefoet al. [l1 was recorded when the specimen was loaded for 1 min. Then, the pressure was unloaded. The value l2 of the gauge was recorded when the specimen was unloaded for 0.5 min. Then, the next pressure was subsequently applied. This process was repeated step by step until all stages were finished. The resilient modulus (E) can be computed by the following equations.P is the maximum pressure, MPa; H is the height of specimen, mm; and l, Baghini et al.  indicateet al. , a maximet al. [Pozo-Antonio  and Idiaet al.  indicateet al. . Two glaim, im−1 are the masses of specimen on the ith day and the (i − 1)th day, respectively, g; pm is the dry mass of specimen, g; δi is the drying shrinkage deformation on the ith day, mm; i,jX and iX−1j, are the values of the jth gauge on the ith day and (i − 1)th day, mm; εi is the drying shrinkage strain on the ith day, %; l is the initial length of specimen before test, mm; αdi is the shrinkage coefficient on the ith day, %.In this study, the drying shrinkage coefficient was employed to evaluate the drying shrinkage performance. It can be calculated by the following equations:et al. [et al. [FRI is the indicator that represents the frost resistance—a high value means a good frost resistance. FTR is the unconfined compressive strength of specimen after five freeze–thaw cycles, MPa; and cR is the unconfined compressive strength of control specimen, MPa.Kelestemur et al.  and Jafa [et al.  have ind [et al. . These set al. [Palomo et al.  indicateet al. [In order to save the construction cost, lime was firstly used to stabilize the boron waste alone. The feasibility of the lime-stabilized boron waste (LSB) for the road base was investigated. The optimum moisture content (OMC) and maximum dry density (MDD) for every proportion were determined. Kweon et al.  indicateet al. [It can be seen from et al. . The uncet al. . For theAs shown in et al. [et al. [et al. [Balen et al.  proposed [et al.  pointed  [et al.  proposed [et al. . The comet al. [et al. [et al. [et al. [et al. [et al. [As listed in et al.  and Moda [et al. . Edeh et [et al.  proposed [et al.  also tho [et al.  thought  [et al.  also ind [et al. . In otheUnconfined compressive strength (UCS) is a common mechanical property, which has been used to determine the proportion of stabilized mixture. Therefore, the UCS’s of the stabilized mixtures were investigated. Three curing periods at 7, 28, and 90 days were selected for the experiment. Results are shown in et al. [et al. [It can be seen from et al.  and Taha [et al.  and is a [et al. . Therefoet al. [As shown in et al.  pointed et al. [et al. [For a cementitious composite, Idiart et al.  indicate [et al.  proposedet al. [E) was obtained by a uniaxial compression test. The indirect tensile strength (ITS) and ultimate tensile strain (UTS) were calculated by Equations (1) and (2), respectively. Results are listed in As shown in et al.  indicateet al. . TherefoE) of LCBS increased with the increase in boron waste. For LCBA, the resilient modulus decreased with the increase in boron waste. Their trends are different. This is caused by the difference in the elastic properties of aggregate, boron waste, and soil. ITS and UTS decrease with the increase in boron waste. This means that boron waste has an adverse effect on the tensile properties. Besides, ITS of LCBA-1 is the highest among all of the mixtures. The ultimate tensile strain (UTS) of LCBS-1 is the highest. UTS of LCBA-1 decreases by 17.0% compared with that of LCBS-1. It seems that the decline in UTS is adverse to drying shrinkage, but the shrinkage coefficient of LCBA-1 is less than that of LCBS-1. Therefore, LCBA-1 may be a good proportion on the whole.It can be found from FTIR spectra of the lime-stabilized matrix and the lime–cement-stabilized matrix were recorded with a Nexus 6700 spectrometer  in order to reveal their chemical reactions. Results of the FTIR test are shown in −1 indicates the formation of MgCO3. The bonds around 1000 cm−1 correspond to Si–O and Al–O tension bonds. They are the characteristic bonds of the alkaline polymer. Bonds around 780 correspond to Si–O–Si bonds. Bonds around 3600 cm−1 indicate the presence of CaO or Ca(OH)2. SiO2 and Al2O3 in boron waste will be activated by CaO (or Ca(OH)2), and C–S–H and CaO (or Ca(OH)2) will form the C–S–H gel structure. The reaction equations can be written as follows:As shown in et al. [However, Palomo et al.  proposed1.The unconfined compressive strength (UCS) of lime-stabilized boron waste (LSB) meets the strength requirements of road base when lime content is greater than 8%. Its frost resistance is very poor. Thus, lime-stabilized boron waste can only be used in non-frozen regions.2.A lime–cement-stabilized boron waste mixture has higher compressive and tensile strengths than those of a lime-stabilized one. Drying shrinkage coefficients of lime–cement-stabilized boron waste mixtures are all less than those of lime-stabilized boron waste. Fillers such as soil and aggregate improve drying shrinkage. It is suggested that soil and aggregate are properly added in order to reduce the drying shrinkage coefficient. Frost resistance indexes of lime–cement-stabilized boron waste mixtures are all greater than 70%. Therefore, a lime–cement-stabilized boron waste mixture is suggested for frozen regions. According to the results of this study, the proportion of LCBA-1 is the most suitable for road base.3.2 and Al2O3 in boron waste is activated by CaO (or Ca(OH)2). Boron was not activated by lime and cement. Therefore, boron waste can be reused as filler in road base in order to solve its effect on the environment.The hydration process of lime and cement formed the strength of a stabilized mixture. SiOBoron waste was reused in road base in this study. The performances of lime and lime–cement-stabilized boron waste mixtures were investigated by various experimental methods. Some conclusions can be drawn based on the above analysis. They are as follows:In summary, boron waste can be used to construct the road base directly. Cement should be used to stabilize the mixture in order to enhance its mechanical strength and frost resistance. The use of boron waste in road base will effectively reduce its impact on the environment. In the future, a trial section of road base should be constructed to verify its serviceability for different regions.


Supracondylar fractures are common in children and are associated with significant morbidity.The purpose of our study was to assess and compare the clinical and radiological outcome of management of supracondylar fractures by both wire configurations, along with identifying factors that predispose to complications.We retrospectively reviewed all paediatric cases admitted with a supracondylar fracture over a five year period. We reviewed case notes, theatre records and radiographs to determine the age of the patient, classification of fracture, treatment method, delay to theatre, duration of surgery, wire configuration, Baumann´s angle, radiocapitellar alignment, anterior humeral alignment and complications.During the five year period we admitted 132 patients and complete notes were available for 123 patients for analyses. For all the patients managed with wire stabilisation 23% developed complications, including 13% with significant complications including nerve injuries and fracture displacements. All five nerve injuries had crossed wires, whereas all for fracture displacements had lateral wires. Baumann´s angle was 76.7 degrees in the group with no complication and 72.2 degrees in the significant complication group (p=0.02). Radiocapitellar line and anterior humeral line were not satisfactory in 5% and 15% of the group with no complications, and 17% and 33% of the group with significant complications.We found more complications in lateral pinning configurations, although all nerve injuries were in patients with crossed wire configurations. The factors we believe are associated with a higher likelihood of complications are inadequate post-operative radiological appearance. The iSupracondylar fractures are commonly classified based on the Gartland system of classification, where they are divided into three types; Type I being non-displaced, Type II being displaced but with an intact posterior cortex and Type III being displaced and without any cortical contact , althoug1).Along with a posterior fat pad sign in Type I fractures, three radiographic parameters used to evaluate a supracondylar fracture are the Baumann’s angle, anterior humeral line and radiocapitellar line . The Bau2). Both have advantages and disadvantages. The crossed wire configuration is biomechanically more stable, especially when resisting axial forces [et al. [et al. [Percutaneous K-wiring is the most widely advocated method to stabilise displaced supracondylar fractures after reduction. There is no clear consensus on the configuration of K-wiring. Commonly used configurations include a crossed configuration with a medial and a lateral K-wire, and lateral configuration with two lateral K-wires  l forces -20. Brau [et al.  conducte [et al. , 21. Bra [et al.  also rep [et al. .The purpose of our study was to assess and compare the clinical and radiological outcome of management of supracondylar fractures by both wire configurations, along with identifying factors that predispose to complications.2We retrospectively reviewed all supracondylar fractures in children between the ages of 2-15 years old that were admitted to our unit between November 2009 and November 2014. We carried out a review of case notes and theatre records to determine the age of the patient, time to operation theatre and duration of surgery. We carried out a review of patients’ radiographs to determine the type of Gartland fracture, Baumann’s angle, radiocapitellar alignment and anterior humeral line post-intervention/surgery. Radiographs were also used to determine the type of wire configuration used. We performed students t-test for statistical analyses and a p value of <0.05 was considered significant.3There were 132 patients with Gartland type II and type II fractures admitted to out unit over the five year period and complete notes were available for 123 patients. Out of 123 patients, 12 were managed nonoperatively, and 13 were managed with a manipulation under anaesthesia. None of these patients had any complications. All the remaining 98 patients were treated with K-wiring, either crossed or lateral. They had a mean age of 6.1 years (SD 2.6 years). These included 61 type II and 37 type III fractures. Fifty-nine patients were managed with crossed K-wires and 39 were managed with lateral K-wires. The ages and fracture types were not significantly different between the two wire configuration groups.Out of these patients managed with wire stabilisation 23% (22 patients) developed complications, including 13% (13 patients) with significant complications including nerve injuries (five patients) and fracture displacements (four patients). Out of the five nerve palsies, two were ulnar nerve palsies, two were radial nerve palsies, and one was a median nerve palsy. The mean age, classification, time to theatre and duration of surgery were not significantly different between the patients with and without complications (p > 0.05). The rate of complications was not different between the two groups; 33% in lateral wire configuration compared with 26% in those treated with crossed wires. Five of the significant complication patients had lateral wire configuration whereas the other eight had crossed wires. All five nerve injuries had crossed wire configuration, whereas all four fracture displacements had lateral wire confirmation. The mean Baumann’s angle was 76.7 degrees in the group with no complication and 72.2 degrees in the significant complication group (p=0.02). The radiocapitellar line and anterior humeral line were not satisfactory in 5% and 15% of the group with no complications, and 17% and 33% of the group with significant complications.4et al. [et al. [et al. [A systematic review  in 2012 et al.  patients [et al. . The aut [et al.  looking et al. [et al. [et al. made an incision over the medial epicondyle. Gatson et al.. treated four crossovers as intention-to-treat whereas Kocher et al. excluded crossovers. Both studies looked at ulnar nerve injuries and changes in Baumann’s angle and humerocapitellar angle. Only Gatson et al. looked at range of movement and loss of carrying angle. Neither study found a significant difference in the clinical or radiological parameters between the two wire configurations. Although the definition for loss of reduction varied between the two studies, neither identified a difference between the wire configurations in relation to loss of reduction. Although Kocher et al. did not report any nerve injuries, Gaston et al. reported two cases with the crossed configuration. They report one case of ‘tenting of the nerve’ with incomplete recovery at three months follow-up, and one case of ‘pin indenting the nerve’ at 90 degrees of elbow flexion with complete recovery at three months.Two further randomised controlled trials were performed by Gaston et al.  and Koch [et al.  looking Our results suggest that both wire configuration patters are valid but the complication profile varies. In our study fracture displacement was seen only with lateral wiring, and nerve injuries only seen with crossed wires. Our results support those of biomechanical studies , 21, 28 Our study has limitations. It is a retrospective study where the procedure as carried out by a number of different surgeons. We did not look at the three lateral wire configuration or K-wire sizes. There is some evidence that three lateral wires produce a more stable construct than two lateral wires , 28, andWe found more complications in lateral pinning configurations, although all nerve injuries were in patients with crossed wire configurations. The factors we believe are associated with a higher likelihood of complications are inadequate post-operative radiological appearance. We suggest that future randomised controlled trials are sufficiently powered with larger patient numbers to detect significant differences in clinical and radiological outcomes.


PLOS One reviewing process. The reality is that Pilling et al. (PLoS One 8:e77193, PLOS One and within the published EFSA Evaluation of Thiamethoxam.The published Commentary by Hoppe et al. (Environ Sci Eur 27–28,  PLOS One insisted that they be combined into a single manuscript. In the end, the final Pilling et al.’s [http://registerofquestions.efsa.europa.eu/roqFrontend/outputLoader?output=ON-3067). A detailed authors’ response to each of the points raised by Hoppe et al. [In response to the increasing call for industry to be more transparent, Syngenta took the decision to publish the pivotal honeybee field studies that supported the honey bee safety of thiamethoxam. These field studies included 12 separate pollen and nectar field residue trials and five long-term (4 consecutive years) field effects studies on honeybees carried out in four geographically widespread locations in France. The primary purpose of these field trials was to investigate and test the bee safety of thiamethoxam. As such these studies have to be carefully designed to avoid as far as is possible any other potential confounding factors, e.g. exposure to other insecticides. In response to the criticism of Hoppe et al. 2105) that the “design and adherence to the protocol was described inadequately” and that it was “doubtful whether the study was implemented in a traceable way”, it should be noted that these studies also have to comply with the strict, legally enforceable, quality control requirements of Good Laboratory Practice  to ensure a viable crop.This quoted figure of 70 % total colonies lost in Hoppe et al.  is complHoppe et al.  state thAs can be seen from Table Hoppe et al.  compare Hoppe et al.  criticisPLOS One’s decision to publish Pilling et al.’s [PLOS One carried out a second additional review of this paper, by a member of the PLOS One editorial board. Once again this paper was accepted as stands (see link to detailed comments from this second review http://www.plosone.org/annotation/listThread.action?root=82356). One of the comments made by the editor during this second review was “The effort was comprehensive and seems honestly described.”Hoppe et al.  questionet al.’s  paper, bet al.’s  paper waWe contend that the alleged deficiencies claimed by Hoppe et al.  to under


Phaseolus vulgaris L.) is a favored food legume with a small sequenced genome (514 Mb) and n = 11 chromosomes. The goal of this study was to describe R and LD in the common bean genome using a 768-marker array of single nucleotide polymorphisms (SNP) based on Trans-legume Orthologous Group (TOG) genes along with an advanced-generation Recombinant Inbred Line reference mapping population  and an internationally available diversity panel. A whole genome genetic map was created that covered all eleven linkage groups (LG). The LGs were linked to the physical map by sequence data of the TOGs compared to each chromosome sequence of common bean. The genetic map length in total was smaller than for previous maps reflecting the precision of allele calling and mapping with SNP technology as well as the use of gene-based markers. A total of 91.4% of TOG markers had singleton hits with annotated Pv genes and all mapped outside of regions of resistance gene clusters. LD levels were found to be stronger within the Mesoamerican genepool and decay more rapidly within the Andean genepool. The recombination rate across the genome was 2.13 cM / Mb but R was found to be highly repressed around centromeres and frequent outside peri-centromeric regions. These results have important implications for association and genetic mapping or crop improvement in common bean.Recombination (R) rate and linkage disequilibrium (LD) analyses are the basis for plant breeding. These vary by breeding system, by generation of inbreeding or outcrossing and by region in the chromosome. Common bean ( Common bean is an important food legume with interesting genetics that is also a good protein and micronutrient source for many consumers around the world . The croLinkage disequilibrium (LD) analysis is related to population genetics and has been the basis for genetic and association mapping done in common beans and other inbreeding crops, especially with advanced generation populations . LD is tLD is influenced, among other factors, by the rate of chromosomal recombination (R) within a species across multiple generations of inbreeding or cross-breeding and depends on the mating system of the plant and the location within the genome . Thus, rApart from these issues, epistatic interactions may create non-random associations among unlinked loci, and genomic differentiation between subspecies can limit LD decay . Since RSingle nucleotide polymorphism (SNP) markers have been developed over the past six years for common beans. The first SNPs to be developed for the crop were designed based on amplicon re-sequencing of mapping parents ,16 conseet al. [et al. [et al. [et al. [et al. [et al. [LD analysis and evaluations of R can be organized based on a single chromosomal region, various genomic regions or the entire genome, especially when working with SNP markers. For example, SNP markers have been useful in locus specific studies ,23–26 oret al.  were sho [et al. . However [et al.  or Song  [et al. , have a  [et al.  used a R [et al.  used an  [et al.  has not The goals of this study were 1) to provide the genetic and physical map locations for the gene-based SNP assays developed by Blair et al.  which aret al. [et al. [et al. [et al. [et al. [et al. [2 to a fine powder, which was mixed with extraction buffer and incubated at 65 C in a 15 mL Falcon tube. Protein removal was accomplished using two chloroform–isoamyl alchohol extractions at 1:1 ratio, which were shaken with the tissue homogenate and centrifuged at 10,000 rpm, removing the upper aqueous layer for DNA precipitation. The resulting purified DNA was quantified on a Hoefer DyNA Quant 2000 fluorometer and diluted to a standard concentration (200 ng/μl) for use in SNP marker evaluation.The plant material used in this study were 1) a recombinant inbred lines (RIL) of the inter-genepool, Andean x Mesoamerican genetic mapping population BAT93 x Jalo EEP558, described as a core mapping population by Freyre et al.  and used [et al.  and the  [et al.  and 2) a [et al. . These w [et al.  that rep [et al. . The pla [et al. , which iet al. [http://dnatech.genomecenter.ucdavis.edu/). SNP genotyping calls were made with Bead-studio software package .The SNP markers used for this study were from the legume trans-legume orthologous gene (TOG) series made for common bean from the sequencing of BAT93 and Jalo EEP558 parental genotypes as described in Blair et al. . The maret al. [et al. [11).Alleles for the population were then used for genetic mapping carried out in Mapdisto v. 2.0  assuminget al.  and simp [et al.  were plablast.ncbi.nlm.nih.gov) search using default parameters for significance. The query sequence included the 120 bp of flanking sequences, or 60 bp on both left and right sides, of each of the common bean SNP markers compared to the chromosomal sequences available for Phaseolus vulgaris [phytozome.jgi.doe.gov). The most homologous physical position of each SNP amplicon sequence was estimated based on the lowest E-value hit found by the similarity search and recorded in Mega base pairs (Mb). Multiple matches were not considered for the TOG markers. The physical positions of the SNP markers were used in the construction of a customized comparative map for all 11 chromosomes carried out with the software R  showing physical (Mb) and genetic (cM) distances. Chromosomal identity and the orientation per chromosome were based on the physical map. After collecting mapping information on both scales, scatter plots were created with R software to analyze the relationship between linkage map distance  and physical distance  for each chromosome. Polynomial line-fitting was used to determine the points of inflection and flattening in the curves fit to each of the chromosomal plots; with these indicating suppressed recombination typical of the centromeres and peri-centromeric (pCENR) regions as described in Bhakta et al. [et al. [The genetic map from the methodology described above was predominantly made of TOG markers and was aligned with the physical map through sequence comparisons through a nucleotide BLASTn   markers, which are similar to the COS (conserved orthologous sequence) markers that have been useful in Solanaceous plant species and advocated by Lee et al. [Phaseolus vulgaris G19833 genome assembly, Phytozome v1.0. The overlap (intersection) between markers and genes was calculated using the bedtools “intersect” function [As a core set of SNPs, the highly conserved markers from this study are useful because of their association with orthologous loci across Expressed Sequence Tags from the transcriptomes of e et al. . To furtfunction .a priori criteria of stratification using STRUCTURE 2.3.2 [et al. [Genepools based on population structure were determined with non RE 2.3.2 , a burn- [et al. . Polymor2) between all pairs of markers with the software package TASSEL 2.1 [2 capture different aspects of the gametic associations [2 estimate were obtained with a two-sided Fisher’s exact test as done in the same program.The overall LD was estimated by calculating the square value of correlation coefficient , in several contexts. The TOG-based map is viewable using CMap v. 1.01 [Phaseolus vulgaris v1.0 GBrowse genome viewer, at https://legumeinfo.org/genomes/gbrowse/Pv1.0. Underlying data files are at the LIS database: https://legumeinfo.org/data/public/Phaseolus_vulgaris/mixed.map1.7PMp/.The genotypes used are in  v. 1.01  with maret al. [et al. [https://legumeinfo.org/data/public/Phaseolus_vulgaris/mixed.map1.7PMp/.The SNP based BAT93 x JaloEEP558 genetic map was constructed with all the single copy TOG markers from Blair et al.  and was et al. , as wellet al. , resulti [et al. . Groups et al. [Of the 812 genetic markers in the final map, all 768 SNP markers were tested for physical mapping to the common bean genome sequence from Schmutz et al. . In totaThe cM / Mb scatter plots for all common bean chromosomes  and for Phaseolus vulgaris genome. Meanwhile 66 markers had possible multiple hits under the parameter of difference of e-10 from the best hit to next best hit and an E-value threshold of 1e-30. At the 1e-40 level, 497 TOG markers were exclusive hits to one gene; while in a total number of cases two TOG markers corresponded to the same gene. No difficulties were found searching for the BAT93/Jalo EEP558 derived TOG markers against the genome assembly for G19833, which was the genotype used in the sequencing project of Schmutz et al. [-30. The results of gene correspondences is given in Of the 768 new TOG markers, the majority, a total of 702 sequences had highly significant singleton hits to predicted or actual genes in the I on linkage group b02d, at the end of the short arm of chromosome Pv02 was flanked at 0.9 cM by three new SNP markers, namely TOG961744_119, TOG906764_834 and TOG906764_376 in this highly recombinogenic and evolutionarily active region that has been amply characterized for the understanding of the necrotic response to BCMNV strains of bean common mosaic virus [Since the TOG markers were well distributed, it was important to note that some of the new SNPs could be useful in substituting phenotypic or legacy molecular markers. In the case of phenotypically useful markers the virus resistance gene called dominant ic virus .Phs) which influences seed size and where TOG897715_56 and TOG897715_587 were genetically linked at 1.7 cM. Several examples of SNPs linked to isozymes mapped by Freyre et al. [Aco2), chalcone synthase (ChS), chitanase (Chi) and diaphorase (Diap); while the substitution of RFLP markers by SNPs is self-evident.Another well-characterized gene with new flanking SNP markers was the locus for phaseolin protein 2. Distinction of linkage groups by linkage disequilibrium blocks was only achievable in the analysis carried out within the Andean genepool. On the other hand, inter-chromosomal linkage disequilibrium was more prevalent within the Mesoamerican genepool likely due to its extensive race substructure. When genepool structure was not accounted for, genome-wide linkage disequilibrium was notoriously widespread and did not decay with genetic distance. To study the relationship of R and LD, respectively, with gene density (2 (%) and recombination rate (cM/bp) value in that window. The relationship was significant in both cases with P = 0.031 and P<0.0001, respectively. Therefore, we can conclude that where the gene density was higher, the recombination rate and r2 were higher. This indicated that LD decay was higher in windows that were gene-rich compared to those windows that were gene-poor.Overall LD measured as romosomes  and at eomosomes . As D′ i density , we condThe TOG based SNPs were useful for creating a saturated genetic map for the BAT93 x Jalo EEP558 population, which has been widely used in previous studies as a reference population . The SNPet al. [et al. [In both the work of Blair et al.  and Hyte [et al. , the dis [et al. , but the [et al. ,44,45. GPhaseolus vulgaris was useful for linkage group to chromosome identification and therefore marker orientation. For example the isozyme or phenotypic markers for BCMV resistance (I gene) and enzyme Chs mapped to the correct locations on Pv2 as did the loci for isozymes Aco2 and Diap on Pv5 and the locus for Chi and the seed protein phaseolin (Phs) on Pv7 according to original mapping [et al. [In our study, observations from the refined genetic map showed the TOG markers were well distributed across and within all the linkage groups with even genetic and physical distances between most markers with the exception of those at chromosome ends. Another observation was that the genetic map was smaller overall than previous maps. This could be explained by the fact that some of the SNP markers were grouped in blocks, making for a more condensed genetic map. Meanwhile, the full genome sequence of  mapping , for thi [et al.  for the Χ2 = P≥0.05). We analyzed the SNP distribution further in two ways: 1) by comparing linkage groups and 2) by comparing regions within the physical map for each chromosome.In the final genetic mapping the number of SNP markers varied between linkage groups. The number of total markers per linkage group ranged up to 145 for Pv2 (49.0 Mb) with more than 110 markers on Pv1 (52.2 Mb in length) and Pv3 (52.3 Mb). Meanwhile, linkage groups Pv4 (45.8 Mb), Pv5 (40.7 Mb) and Pv10 (42.2 Mb) were low in SNP marker saturation with 22, 40 and 27 TOGs, respectively. The remaining linkage groups had similar numbers of markers ranging from 61 on Pv11 (50.2 Mb) to 88 on Pv7 (51.7 Mb). A chi-square test (significance P≤0.05) showed that the distribution was not equal between linkage groups for the SNP markers. In contrast the RFLP and SSR markers were more evenly distributed as anchors across all linkage groups  was telocentric, as predicted by Bhakta et al.  due to tet al. [et al. [et al. [et al. [Our genetic mapping results compared favorably with those of the common bean genome sequencing and re-sequencing study of Schmutz et al.  or the n [et al. . In thos [et al.  was corr [et al.  but to a [et al. , perhaps [et al.  was modeet al. [et al. [I gene for virus resistance on Pv02.In summary regarding the linkage analysis, we presented a saturated genetic map with well characterized SNPs on a reference population for common bean, derived from the cross BAT93 x Jalo EEP558, and we used this map for estimating recombination rates and LD decay across the genome. This genetic map has been useful in determining recombination rates among many types of markers. As a core mapping population, the BAT93 x JaloEEP 558 set had many comparable markers with other highly saturated genetic maps such as those of Córdoba et al.  and Gale [et al. . As concet al. [LD estimates were made for the same SNP markers that were genetically mapped above and for both Andean and Mesoamerican genotypes from Blair et al. , which aet al. ,11. The et al. [et al. [A further result of our study was that LD was stronger and decayed slower within the Mesoamerican genepool, likely due to its more extensive race substructure. This is interesting because a bottleneck for the Andean beans, as has been speculated by arguing a Mesoamerican origin of the common bean , would iet al.  and by Ket al. . The Braet al.  found si [et al. . The abi [et al. , sub-spe [et al. . Domesti [et al. .A final point for our study was that the rate of LD decay was low when marker associations were studied within linkage groups and for each genepool separately. More focused studies of LD at certain loci have shown that LD decay was moderate and variable around drought tolerance genes –26 acrosWithin Andean LD analysis is likely to show higher recombination than within Mesoamerican genepool mapping due to less population structure and potentially higher polymorphism. LD rates have been shown to affect the potential for QTL and association mapping of traits in common bean. Given the higher within-Andean LD, initial successes with genome wide association have been for Andean studies ,53,54. MS1 Table(DOCX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file.S1 Fig(PDF)Click here for additional data file.S2 Fig(PDF)Click here for additional data file.S3 Fig(PDF)Click here for additional data file.S4 Fig(PDF)Click here for additional data file.S5 Fig(PDF)Click here for additional data file.S6 Fig(PDF)Click here for additional data file.S7 Fig(PDF)Click here for additional data file.


Although medication is generally avoided wherever possible during pregnancy, pharmacotherapy is required for the treatment of pregnancy associated hypertension, which remains a leading cause of maternal and fetal morbidity and mortality. The long-term effects to the child of in-utero exposure to antihypertensive agents remains largely unknown.The aim of this study was to systematically review published studies on adverse outcomes to the child associated with in-utero exposure to antihypertensive medications.OVID, Scopus, EBSCO Collections, the Cochrane Library, and Web of Science databases were searched for relevant publications published between January 1950 and October 2016 and a total of 688 potentially eligible studies were identified.Following review, 47 primary studies were eligible for inclusion. The Critical Appraisal Skills Programme checklist was used to assess study quality. Five studies were of excellent quality; the remainder were either mediocre or poor. Increased risk of low birth weight, low size for gestational age, preterm birth, and congenital defects following in-utero exposure to all antihypertensive agents were identified. Two studies reported an increased risk of attention deficit hyperactivity disorder following exposure to labetalol, and an increased risk of sleep disorders following exposure to methyldopa and clonidine.The current systematic review demonstrates a paucity of relevant published high-quality studies. A small number of studies suggest possible increased risk of adverse child health outcomes; however, most published studies have methodological weaknesses and/or lacked statistical power thus preventing any firm conclusions being drawn. Hypertensive disorders of pregnancy have been identified as the leading cause of maternal death in industrialized countries, complicating approximately 7% of pregnancies ,2.Prior to pregnancy, approximately 3% of women have an existing diagnosis of hypertension of whom a quarter will go on to develop preeclampsia ,3. A furConsequently, there has been a move to treat hypertensive women of childbearing age more aggressively in an attempt to normalize blood pressure (BP) prior to and during pregnancy and reduce the occurrence of preeclampsia . DespiteAlthough in-utero exposure to any medication during the first trimester is associated with the highest risk of teratogenic malformations, exposure during the second and third trimesters has also been linked to functional and behavioural abnormalities, which may not be immediately apparent –9.Although the fetogenic and teratogenic risks associated with angiotensin-converting enzyme inhibitors (ACEi) and AT1 blockers (ARB) use during pregnancy are well recognized  the potein utero to one or more antihypertensive medications compared with unexposed children.To determine whether in-utero exposure to antihypertensive medication is associated with adverse child outcomes, we reviewed primary literature reporting child outcomes following exposure A systematic review protocol was designed as per standard Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines . The datThe reference lists from eligible papers were scanned for other relevant studies. The search was limited to English and German language articles. Narrative and systematic reviews (with no synthesis of data), studies published only as abstracts, letters, or conference proceedings, discussion papers, animal studies, and editorials were excluded.Initial screening of titles was carried out to identify potentially relevant studies, followed by screening of abstracts and then by full paper review. All titles and abstracts were independently evaluated by two reviewers (M.S. and J.S.M.) for consistency of inclusion/exclusion.Quality assessments were conducted by two independent reviewers (C.F. and M.S.) using a modified version of the Critical Appraisal Skills Programme quality assessment tool for randomized controlled trials , case–cohttp://www.crd.york.ac.uk/prospero/[http://www.prisma-statement.org/.The current review was registered with the International Prospective Register of Systematic Reviews: prospero/. The PRIThe database searches retrieved 816 citations. Removal of duplicates followed by review of titles and abstracts yielded 47 relevant studies. The PRISMA flowchart  illustrates the number of titles, abstracts, and full papers excluded.In total, 22 studies were conducted in Europe; 16 in North America, three in Asia, one in central America, and one in Africa. Four of the 47 studies were multinational, three of which were across continents . Of thesN.Descriptive tables Tables –6 showinStudies were categorized according to study design and quality and bias assessed accordingly . No study achieved the maximum score for quality, five were graded as excellent ,31,33,51For clarity and ease of interpretation, the results of the eligible studies have been reported, according to the drug of exposure, as early (birth to 1 month) and long-term (>1 month) outcomes.Five case–control studies –28,30,31et al.[et al.[In the cohort study of 1418 women, rated as excellent, Lennestål et al. reportedl.[et al. also repet al.[In the case–control study rated as excellent, studying major congenital anomalies , and being small for gestational age , Nakhai-Pour et al. reportedet al.[P < 0.0001) and increased odds of low birth weight  [Banhidy et al. reported1.8–2.7) . However1.8–2.7) .et al.[Caton et al. reportedet al.[In a second case–control study rated as mediocre, by Caton et al., 5021 caet al.[et al.[Van Gelder et al. reportedl.[et al. reportedAny hypertensive summary. In summary, although seven studies reported increased odds of preterm birth and low birth weight in treated mothers, particularly those with chronic hypertension and four studies, a possible increase in the incidence of congenital malformations study designs did not allow assessment of the relative importance of hypertension vs. exposure to antihypertensive patients. The ORs for perinatal mortality, preterm birth, and congenital cardiovascular defects are reported as Forest plots in Supplementary Figs. 2–4.One case–control study , 16 cohoet al.[In a cohort study of 1063 238 pregnant women, rated as excellent, Lennestål et al. reportedet al. also repet al.[In a cohort study of 100 029 women, also rated as excellent, Orbach et al. reportedet al. also repHowever, there was also increased odds of preterm birth , being small for gestational age , and intrauterine growth restriction , for offspring exposed to untreated maternal hypertension in-utero compared with the normotensive untreated reference group. The authors  did not et al.[In a mediocre cohort of 911 685 women, Meidahl Petersen et al., reporteet al. also repet al.[Ray et al., in a stet al.[P < 0.001), low birth weight , and being small for gestational age  following atenolol exposure compared with an untreated hypertensive comparison group, with the largest risk of being small for gestational age following exposure to atenolol during early pregnancy .In a mediocre cohort study of 312 women, Lydakis et al. reportedet al.[In a cohort study of 1223 women rated as poor, Xie et al. reportedet al.[P = 0.25), being small for gestational age , or being born preterm , in 109 offspring, 55 of whom were exposed in utero to labetalol [P = 0.02) associated with labetalol. In a poor cohort study of 491 women, Bayliss et al.[P < 0.01), and lower ponderal index  following in-utero exposure to atenolol in the first trimester of pregnancy, compared with untreated hypertensive women.In a cohort study rated as poor, investigating the possible effects of labetalol for the treatment of severe preeclampsia, Heida et al. reportedabetalol . The autss et al. reportedet al.[P < 0.001) in a cohort study, rated as poor, of 398 women. The authors [3 × 104; P = 0.001).Lip et al. reportedet al.[et al.[Caton et al., in a mel.[et al. reportedet al.[et al.[P < 0.01). In a poor cohort study investigating the effects of labetalol on 129 offspring, Munshi et al.[P < 0.01); however, there was no significant difference in the incidence of intrauterine growth retardation or birth asphyxia.In a mediocre RCT of 100 women assessing fetal head circumference and placental weight, Fidler et al. observedl.[et al. reportedhi et al. reportedet al.[N = 11), compared with a comparison group of unexposed, normotensive women (N = 11). There was no difference in heart rate (HR), respiratory rate or palmar sweating between the two treatment groups; however, mean SBP in infants was significantly different at 2-h post birth (P < 0.05). This difference disappeared after 72 h.In a poor cohort study of 22 women, Macpherson et al. reportedet al.[et al.[P < 0.05).In an RCT of 176 women rated as mediocre, Plouin et al. reportedl.[et al. reportedet al.[P = 0.02).Lieberman et al. reportedet al.[et al.[P = 0.008) and hypotension . No differences in birthweight, intrauterine growth restriction, hypoglycemia, respiratory distress syndrome, 1 and 5 min Apgar scores, or NICU admissions were reported between the two treatment groups.In a mediocre RCT of 263 women with mild-to-moderate chronic hypertension, Sibai et al. reportedl.[et al. reportedet al.[In a poor cohort study to investigate functional development of the fetal central nervous system, involving 117 women, 21 of whom were treated with nifedipine or labetalol for pregnancy-induced hypertension, Gazzolo et al. reportedBeta-blocker summary: In summary, exposure to beta-blockers in the setting of hypertension, was associated with an increased risk of preterm birth, being small for gestational age, increased perinatal mortality and low birth weight; however, there was limited evidence to suggest that this increased risk was due to the medication rather than the underlying hypertensive disease, with only six of 13 studies reporting increased risk controlling for underlying hypertension. There was also some evidence of an increased risk of cardiovascular malformations; however, relevant studies did not include an untreated comparison group [on group ,29. Threon group ,44,53, wThree RCTS ,62,63, oet al.[In the one study rated as excellent, Bateman et al. reportedin utero to nifedipine and offspring without in-utero antihypertensive exposure.Gruppo , in an Ret al.[P = 0.003) compared with normotensive untreated women in a poor cohort study of 156 women. The authors [P = 0.08), or congenital malformations  among offspring with in-utero exposure to calcium channel blockers.Magee et al. reported authors  reportedet al.[P = 0.007), miscarriages  associated with in-utero exposure to calcium channel blockers compared with an untreated normotensive group. The authors [P = 0.10) with in-utero exposure to calcium channel blockers compared with an untreated normotensive group.Weber-Schoendorfer et al., in a co authors  reportedet al.[P = 0.03).Hall et al. reportedet al.[In a mediocre case–control study of 54 016 women, involving 22 865 cases taken from the Hungarian Congenital Abnormality Registry and 38 151 controls without malformations, Sorensen et al. reportedet al.[In an RCT of 49 hypertensive women rated as mediocre, Fenakel et al. reportedet al.[et al.[In a mediocre cohort study of 87 407 pregnant women, Davis et al. reportedl.[et al. also repCalcium channel blocker summary. In summary, the results of the calcium channel blocker studies are mixed, with evidence of increased perinatal mortality from two RCTs, increased odds of preterm birth and perinatal mortality from the cohort studies, and evidence of increased malformations from the case–control study. None of the reviewed studies assessed the effect of untreated hypertension on outcomes, thus limiting the interpretation of the data and the relative importance of in-utero exposure rather than underlying disease. The ORs for perinatal mortality, preterm birth, and congenital cardiovascular defects are reported as Forest plots in Supplementary Figs. 2–4.One case–control study  and seveet al.[In a cohort study involving 465 754 offspring, rated as excellent, Li et al. reportedet al. reportedet al.[In a cohort study involving 1063 238 women, rated as excellent, Lennestål et al. reportedet al.[P < 0.001), low birth weight , lower gestational age , and miscarriage  associated with in-utero exposure to an ACEi or ARB, in comparison with a nonteratogen-exposed group. Diav-Citrin et al.[P = 0.838) or congenital malformations .In contrast, in a cohort study rated as poor, comparing ACEi and/or ARB (252 women) therapy vs. other antihypertensive therapy (256 women) or nonteratogenic exposure (495 women), Diav-Citrin et al. reportedin et al. did not et al.[Tabacova et al. reportedet al. also repet al.[P < 0.001), lower birth weight (P < 0.001), and lower gestation ages (P < 0.001) in those exposed to ACEi or ARBs compared with those exposed to other antihypertensive medications and untreated normotensive mothers, in a cohort study of 388 women, rated as poor. The authors [P = 0.99).Moretti et al. reported authors  reportedet al.[et al.[et al.[Caton et al. reportedl.[et al. reportedl.[et al. reportedACEi and ARB summary. In summary, ACEi and ARBs were associated with an increased risk of preterm delivery and miscarriage. The results relating to congenital abnormalities were conflicting with four studies reporting no increased risk and five reporting increased risk. The results, as a whole, are further confounded by the lack of inclusion of an untreated hypertensive group. The ORs for congenital cardiovascular defects are reported as a Forest plot in Supplementary Fig. 4.Two cohort studies ,65 and tet al.[In the only study rated as excellent, Orbach et al. reportedet al.[et al.[P < 0.05). However, there was no difference in IQ or behavior.In a cohort study of 1223 women rated as poor, Xie et al. reportedl.[et al. reportedet al.[et al.[Fidler et al. observedl.[et al. reportedMethyldopa summary. In summary, methyldopa exposure was associated with an increased risk of preterm birth, perinatal mortality, and low birth weight. The RCTs reported no difference among methyldopa exposed offspring in other outcomes such as BP, IQ, behavior and placental weight. An increased risk of intrauterine growth restriction and low Apgar scores was reported by one cohort study. However, the study designs used did not permit any conclusions to be drawn as to the relative importance of in-utero methyldopa exposure vs. the underlying effects of hypertension on the offspring. The ORs for perinatal mortality and preterm birth are reported as Forest plots in Supplementary Figs. 2 and 3.One linkage cohort study  involvinin utero on later childhood development. The RCT [One RCT  and thre The RCT  and two  The RCT ,13 asses The RCT  assessed The RCT ,13,49 an The RCT  quality.et al.[Cockburn et al., in a meet al.[Chan et al., investiet al.[In a mediocre cohort study of 202 patients, Pasker-De Jong et al. investiget al.[in utero, suggesting a possible dose–response relationship.In a poor cohort study of 44 patients, Huisjes et al. reportedThe current study reviewed 47 published studies reporting the effects of in-utero exposure to antihypertensive medication on the fetus and child. Thirty-two of these studies were of poor or mediocre quality, with small study populations, and incomplete adjustment for confounding, and lack of quality.Although there is a widely held view that antihypertensive patients, such as beta-blockers, may be associated with a variety of detrimental fetal outcomes, such as low birth weight or congenital malformations, these beliefs are not based on robust data from appropriately designed and powered studies to conclusively confirm any associations. Furthermore, few studies have investigated the possible long-term outcomes following in-utero exposure to antihypertensive agents. The four studies which have done so have had small study populations, lacked statistical power and reported conflicting results. Although no IQ or developmental differences were reported for methyldopa and labetalol in two studies ,49, sleeThe five studies graded as excellent ,33,51,57et al.[One of the excellent studies assessing any antihypertensive medications by Lennestål et al. reportedThe results of their study seem to indicate a beta-blocker driven association for congenital abnormalities; however, the increased frequency of such abnormalities may have been due to the underlying chronic hypertension. The authors fail to mention the severity of hypertension present in patients, which would impact on treatments used and outcomes of pregnancy. Furthermore, as the authors used only one point of reference to establish drug exposure, it is possible that as pregnancy progressed women may have experienced altered exposure to different antihypertensive patients or other potentially teratogenic agents, possibly confounding the results further. Lastly, because BP was not reported in the first midwife visit at which medication use was established it is not possible to determine whether patients in the untreated comparison group were truly normotensive. Therefore, the reported increase in the prevalence of congenital abnormalities and birth outcomes  reported in this study may be related to underlying, or inadequately controlled, hypertension rather than antihypertensive therapy.et al.[The second excellent study to assess any antihypertensive exposure during pregnancy by Nakhai-Pour et al., reporteThe one study graded as excellent in quality assessing ACEi exposure during pregnancy, reported an increased risk of birth defects when compared with an untreated normotensive group, but no difference when compared with an untreated hypertensive control group . HoweverThe only study to assess calcium channel blocker exposure during the last 30 days of pregnancy rated as excellent, reported no increased risk of seizures compared with an unexposed normotensive comparison group . This stet al.[The fourth study deemed excellent was conducted by Orbach et al., who invTwelve of the 47 studies included in this review were RCTs; 11 of which were reported between 1982 and 2000. The gold standard for study design is an RCT, however, the need for randomization may conflict with the ethical treatment of hypertensive pregnant women and so introduce selection bias in the control groups.et al.[Women included in these RCTs were often hospitalized for severe preeclampsia as reported by Heida et al., which iThe study population size in the RCTs reviewed in this paper was between 25 and 300 patients. As the studies were small scale, they may have lacked statistical power.The majority of reviewed studies were case–control or cohort studies in design, which inherently introduces the issue of confounding from unknown or unmeasured confounders, underlying or poorly controlled hypertension, and the issue of medication adherence. Studies using data from poison centers or teratology centers ,24 may iIt is unclear whether the magnitude of the increased risks, such as birth defects ,57, repoThe results are further confounded by the lack of information describing the types of hypertension treated in women included in the exposed groups. It is likely that the risk profiles and outcomes associated with chronic hypertension, gestational hypertension, and preeclampsia are different; therefore, a knowledge of the underlying condition is important and should be considered and reported. Only 16 of 47 studies reported in this review provided such information, and the majority of these studies assessed beta-blocker exposure.Although it is recognized that the results of cohort studies should be reported as RR, ten of the cohort studies ,51,57,65Treatment of hypertension during pregnancy and assessing the potential risks to the offspring is further complicated by the underlying disease. Untreated hypertension results in preterm birth, low birth weight and increased mortality. It would therefore be reasonable to suggest that poorly controlled hypertension may carry the same risks as untreated hypertension. Case–control and cohort studies reviewed in this paper have failed to determine whether hypertension was appropriately treated and controlled, and failed to assess adherence. Several studies reviewed ,38,45,57Limitations to this review include exclusion of conference abstracts, unpublished studies, studies reported in any language other than English or German, which may have resulted in possible publication bias. However, when the literature search was performed, only two articles were excluded due to language. A further limitation was the wide date-range over which the included studies were published, the majority before 2000. As a result attempted contact with authors was either impractical or unsuccessful.In conclusion, adverse child outcomes such as preterm birth, perinatal mortality, low birth weight, risk of congenital abnormalities, or other detrimental outcomes, following in-utero antihypertensive exposure have been reported in the literature. However, most published studies have had methodological weaknesses and/or lacked statistical power thus preventing any firm conclusions being drawn.Further research in this area is required to ensure that health professionals have sufficient data to treat hypertension during pregnancy to firmly demonstrate a lack of detrimental outcomes.We acknowledge the support from the Farr Institute at Scotland. The Farr Institute at Scotland is supported by a 10-funder consortium: Arthritis Research UK, the British Heart Foundation, Cancer Research UK, the Economic and Social Research Council, the Engineering and Physical Sciences Research Council, the Medical Research Council, the National Institute of Health Research, the National Institute for Social Care and Health Research (Welsh Assembly Government), the Chief Scientist Office , the Wellcome Trust, (MRC Grant No: MR/K007017/1).There are no conflicts of interest.


To monitor the effect of nutrition and pregnancy on oxidative status of animals under the arid condition of South Sinai.Blood samples were taken from two groups of animals: The first group retained in farm and fed on concentrate (high diet) and another group grazing natural forage (low diet). Each group was subdivided into pregnant and non-pregnant animals. Blood samples were assayed for their content of malondialdehyde (MDA), total antioxidant capacity (TAC), catalase (CAT), and superoxide dismutase (SOD) enzymes.MDA level significantly increased in pregnant animals fed either concentrate or grazing low-quality forage and accompanied by a low level of TAC in pregnant grazing animals fed low-quality forage. The activity of CAT decreased in pregnant fed either concentrate or grazing and SOD significant decrease in the pregnant grazing group. These data suggested that the animals might have experienced some degree of oxidative stress and lipid peroxidation and indicating that redox homeostasis was impaired in those pregnant and specially fed on forage rations.Pregnancy constituted the most oxidative stress facing the grazing and concentrated diet feed sheep and goats under arid and saline conditions of Southern Sinai, Egypt. On the other hand, antioxidant enzyme SOD activities were determined in erythrocyte of non-pregnant and late pregnant animals.et al. [et al. [et al. [Lipid peroxidation product as MDA was assayed using the commercially available kit  according to Ohkawa et al.  and expr [et al. , Aebi [1 [et al. , and Nis [et al. , respectData of various parameters were analyzed using computerized software program SPSS version 17.0 using one-way analysis of variance. Comparison of means was carried out by the least square difference. Differences were considered to be significant at p<0.05.The lipid peroxidation, non-enzymatic antioxidant levels, and enzymatic antioxidant activities in pregnant and non-pregnant fed on concentrate or grazing low-quality natural forage are presented in In goats, MDA level significantly increases in both pregnant fed concentrate or grazing than that found in non-pregnant animals fed on concentrate (p<0.01) or grazing (p<0.05). TAC level was lower in pregnant goats fed grazing than the other groups and significantly lower (p<0.01) when comparing with non-pregnant fed concentrate animals. CAT activity significantly decreases in pregnant fed either concentrate or grazing than non-pregnant animals fed on concentrate. Furthermore, CAT activity in grazing pregnant was significant low when comparing with non-pregnant fed concentrate (p<0.001) and pregnant fed concentrate (p<0.05). SOD activity in grazing pregnant was significantly lower value than pregnant fed concentrate (p<0.05) and non-pregnant fed either concentrate (p<0.01) or grazing (p<0.05).In sheep, MDA level in pregnant sheep either fed on concentrate or grazing showed higher level than non-pregnant, but MDA level in grazing pregnant was significant higher (p<0.01) than that found in pregnant animals fed on concentrate and non-pregnant fed on either concentrate or grazing. TAC level in pregnant sheep fed grazing was the lowest level when comparing with other groups but significantly low with non-pregnant fed concentrate (p<0.01) and pregnant animals fed concentrate (p<0.05). CAT activity showed a significant decrease (p<0.05) in either pregnant fed concentrate or grazing than that found in non-pregnant animals fed on concentrate. SOD value in grazing pregnant was significant low than non-pregnant fed concentrate (p<0.01) and grazing non-pregnant (p<0.05).The present study recorded that pregnant goats and sheep were exposed to an increased risk of oxidative stress during pregnancy than non-pregnant groups, by the observed increase of MDA value and decrease of TAC level, CAT, and SOD activities, but the oxidative stress was more observed in pregnant grazing than pregnant fed concentrate sheep and goats. Although nutritional factor is one of the oxidative stress, pregnancy constituted the most oxidative stress facing the animals since oxidative stress index (MDA) of both pregnant groups was increased, but TAC level, CAT, and SOD activities were decreased.Pregnancy is known to be stressful on organisms, which accelerates the production of reactive oxygen species and oxidative stress . The reaIt is well known that high ROS concentrations may lead to oxidative stress and be the cause of many diseases in animals . HoweverLipid peroxidation produces a wide variety of aldehydes, which can be formed as secondary products such as MDA . MDA is Specific biomarkers of lipid peroxidation such as MDA and increased level of MDA are an indication of lipid peroxidation. In our study, MDA level significantly increased in a pregnant animal fed either concentrate or grazing. Similar result obtained in pregnant sheep fed on medium to low-quality forages  and pregOther studies suggested that MDA concentration increased during late pregnancy . On the et al. [The increase in placental progesterone (P4) is accompanied by an augment in blood circulating lipids and MDA, which is a marker of oxidative stress . Mohebbiet al.  reportedet al. , due to et al. [TAC level in our study revealed lower value in a pregnant grazing animal, and this result was in agreement with Amer et al. , who monet al. [et al. [et al. [Our results indicated a decrease in CAT activity in pregnant animals fed either concentrate or grazing, and this result was in agreement with Erisir et al. , who fou [et al.  reported [et al.  recordedet al. [et al. [SOD is well known to be a superoxide radicals’ scavenger which is an essential factor in the protection against free radical damage and is considered the first defense against pro-oxidants . In our et al. , who rec [et al.  found thOur results indicated that changes in the nutritional level of the diet had little effect on animals compared with a factor of pregnancy. In this respect, many findings are consistent with the claim that pregnancy is a state of oxidative stress in ruminants ,6,29-31.et al. [et al. [et al. [It is essential to consider the oxidative stress indicators are affected by nutrition, and modifications of metabolic substrate may affect their biosynthesis and turnover at the tissue level. It has been shown that feeding dairy cows with high levels of starch increases oxidative stress, possibly due to cellular changes related to oxidative phosphorylation . In sheeet al.  recorded [et al.  refer th [et al.  reportedPregnancy constituted oxidative stresses on sheep and goats fed concentrate or grazing. The nutritional level of the diet had very little effect on blood oxidant and antioxidant status.KGMM and MFN designed the experimental study, ARA and ASAS performed oxidant and antioxidants markers assay, whereas KGMM and ARA completed data analysis, revision, and writing of the article. All authors read and approved the final manuscript.


The udder configuration and teat/udder morphology were recorded before milking. Milk samples (100 ml/cow) were collected aseptically. Milk somatic cell counts (SCC) and milk differential leukocyte counts were performed microscopically. Milk leukocytes  were isolated from milk samples by density gradient centrifugation. Phagocytic index (PI) of milk neutrophils and macrophages were evaluated by colorimetric nitro blue tetrazolium assay. Lymphocytes proliferation response was estimated by MTT assay and expressed as stimulation index.In vitro PI of milk neutrophils was found to be significantly (p<0.01) lower in flat teat. In vitro PI of milk macrophages was found to be significantly (p<0.01) lower in the round and flat teats compared to pointed and cylindrical teats.There was a significant (p<0.01) positive correlation between milk SCC with mid teat diameter, teat base diameter and significant (p<0.05) negative correlation between milk SCC and the height of the teat from the ground. Milk SCC was found to be significantly (p<0.01) lower in bowl-shaped udder and higher (p<0.01) in pendulous type. Milk macrophage percentage was positively (p<0.01) correlated with udder circumference. PI of milk neutrophil was negatively (p<0.01) correlation between teat base diameter, and PI of milk macrophages was found to be positively (p<0.01) correlated with teat apex diameter. Both PI of milk neutrophils and macrophages was found to be significantly (p<0.01) lower in the animals having flat and round teat and pendulous type of udder. in vitro activity of milk leukocytes hence facilitates the incidence intramammary infections.Udder risk factors such as teat shape and size, teat to floor distance, udder shape, and size may decrease the  The cr health -5.Association of udder morphology with the occurrence of mastitis has already been established worldwide ,7. Howevin vitro milk leukocyte activity.Therefore, this study is undertaken to shed some light on the association between the udder morphology and The experiments on animals including all procedures of this study were approved by Institutional Animal Ethics Committee (Registration number: 763/03/a/CPCSEA).nd to 4th parity and mid lactations (150-200 days of lactation cycle) were selected from the herd of Eastern Regional Station of ICAR-National Dairy Research Institute, Kalyani, Nadia, and West Bengal, India. All the experimental animals were kept in loose housing system under the routine managemental practices followed in the institute’s herd. All the animals were screened weekly for clinical mastitis throughout the study.A total of 48 high yielding crossbred cows of 2et al. [et al. [The mammary glands of all the animals were analyzed for the presence of any gross lesions. The udder configuration and teat/udder morphology were recorded following Bhutto et al.  before t [et al. . The folTeat length (cm): Average of all four teats.Teat apex diameter (cm): Average of all four teats.Mid teat diameter (cm): Average of all four teats.Teat base diameter (cm): Average of all four teats.Teat shape: Pointed, cylindrical, round, flat.Udder shape: Cup, round, bowl.Udder position: Pendulous, non-pendulous.Composite milk samples (100 ml/cow) from all four quarters were collected into sterile tubes through hand milking for consecutive 3 times with 1 month apart. Teat dipping before the collection was done with an effective teat dip solution (0.5% iodine or 4% hypochlorite) for at least 20-30 s before milking. Then, the teats were carefully scrubbed with a cotton cloth or gauze pad moistened with 70-80% ethyl alcohol.SCC and DLC of milk samples were measured microscopically ,4.Isolation of milk leukocytes - viz., neutrophils, lymphocytes, and macrophages - was performed by density gradient centrifugation -5.In vitro PI of milk neutrophils and macrophages was evaluated by colorimetric nitro blue tetrazolium reductive assay [ve assay .Mitogen-induced milk lymphocyte proliferation response was measured by colorimetric MTT (tetrazolium) assay  and exprin vitro activity of milk leukocytes) was analyzed by Spearman rank order correlation. Effect of teat shape, udder shape, and udder position were analyzed by one-way ANOVA.All analysis was performed using SYSTAT software package. The correlation between udder morphological parameters  and udder immunological parameters  positive correlation between milk SCC with mid teat diameter, teat base diameter and significant (p<0.05) negative correlation between milk SCC and the height of the teat from the ground. There was also a positive correlation between milk SCC and teat length and negative correlation between milk SCC and teat apex diameter though it was nonsignificant.There was no significant correlation between milk DLC and teat morphology. However, milk macrophage percentage was positively (p<0.01) correlated with udder circumference.There was no significant correlation between SI of milk lymphocytes with teat morphology.PI of milk neutrophil was found to be positively correlated with teat apex diameter and height of teat from ground. However, there was a significant (p<0.01) negative correlation between teat base diameter and PI of neutrophils.in vitro PI of milk macrophages with teat apex diameter.PI of milk macrophages was nonsignificantly positively correlated with teat length, mid teat diameter, teat base diameter and height of teat from the ground and negatively correlated with udder circumference. However, there was a significant (p<0.01) positive correlation between in vitro milk leukocytic activity in high yielding crossbred cows have been presented in Effect of teat shape on milk SCC, DLC and In vitro PI of milk neutrophils was found to be significantly (p<0.01) lower in flat teat. In vitro PI of milk macrophages was found to be significantly (p<0.01) lower in the round and flat teats compared to pointed and cylindrical teats.Concanavalin A (Con A) induced milk lymphocyte blastogenic response was found to be unaltered with different teat shape. in vitro milk leukocytic activity in high yielding crossbred cows have been presented in Effect of udder shape on milk SCC, DLC and in vitro milk lymphocyte blastogenic responses were found to be unaltered with different udder shapes. In vitro PI of milk, neutrophil was found to be higher in round shaped udder compared to cup and bowl-shaped udder though the difference was nonsignificant. In vitro PI of milk macrophages was significantly (p<0.01) lower in bowl-shaped udder compared to round and cup-shaped udder.Con A induced in vitro milk leukocytic activity in high yielding crossbred cows have been presented in Effect of udder position on milk SCC, DLC and In vitro PI of milk neutrophil did not differ significantly between different udder positions. However, it was lower in the pendulous type udder. The in vitro PI of milk macrophages was significantly p<0.01 lower in the pendulous type of udder compared to non pendulous type of udder.There were no significant difference in ConA induced milk lymphocyte blastogenic response under different udder positions. et al. [et al. [et al. [et al. [The udder and teats are the first line of defense against intra-mammary infection, and the association between mastitis resistance and several udder type traits have been reviewed by many workers ,8,11. Inet al.  stated t [et al.  reported [et al.  and Sing [et al.  also repin vitro activity of milk leukocytes. Here, we found PI of milk neutrophil and macrophages was positively correlated with teat apex diameter and height of teat from ground which could be explained by the fact that higher activity of milk neutrophils and macrophages in the case of higher teat to ground distance makes the mammary gland more resistance to intramammary infections. Con A induced milk lymphocyte blastogenic response could not be compared as no literature available in this regard.To the best of our knowledge, this is the pioneer study correlating the udder morphology and in vitro milk lymphocyte blastogenic response and PI of milk macrophages were negatively correlated. The earlier reports on the relationship between teat length and mastitis were also contradictory. Some authors suggested longer teats were positively correlated with mastitis incidence [et al. [et al. [et al. [The probability of mastitis occurring varies considerably between different teat shapes, sizes, teat placement, and the morphology of the teat tip . In any ncidence , whereas [et al.  could no [et al.  reported [et al.  stated a [et al. . During  [et al. .in vitro activity of milk leukocytes under different teat shape. In vitro phagocytic activity of milk neutrophils and macrophages were lower in flat type teat compared to round and cylindrical teats. These findings were in accordance with the reports of Slettbakk et al. [et al. [These results suggest that the chances of mastitis are higher if the teat length is shorter and if the teat diameter is greater. There was a significant alteration in the k et al.  and Bhut [et al.  reported [et al. . However [et al.  have alset al. [The udder and teats are the first line of defense against intramammary infection. Udder morphology is very heritable and could serve as a marker trait for selection to reduce mastitis in dairy cattle ,22. Uddeet al.  stated tet al. [et al. [In this present investigation, we found higher activity of milk leukocytes and lower milk SCC in round shaped udder as reported by Saloniemi et al.  indicate [et al.  reportedin vitro activity of milk leukocytes hence facilitates the incidence intramammary infections.The study identified possible udder risk factors for incidence of mastitis such as teat shape and size, teat to floor distance, udder shape and size and found that these factors reduced the JM planned the study. TS recorded the data. PRG and DB provided technical support and helped in data analysis. PKD analyzed the data. JM drafted and revised the manuscript. All authors read and approved the final manuscript.


With the emergence of high-throughput “omics” data, network theory is being increasingly used to analyze biomolecular systems. In particular, proteins rarely act alone, which prompted the arrival of proteomics. “Network proteomics” is just defined as the research area that introduces network theory to investigate biological networks ranging from protein structure networks to protein-protein interaction networks. In addition, the application of network proteomics in biomedical fields has increased significantly. This special issue has collected contributions, not only focusing on the state of the art of methodology in “network proteomics” itself but also focusing on the current status and future direction of their applications in translational medical informatics. Aspergillus oryzae, which would enhance its metabolic network reconstruction. By using the molecular dynamics simulation and principle component analysis, H. Wan et al. investigated the interaction between the last half repeat in TAL effectors and its binding DNA. This work would give a deeper understanding of the recognition mechanism of protein-DNA interactions.This issue starts with discussing the role of protein structure that participates in interactions. N. Raethong et al. developed a strategy to annotateProtein complexes offer detailed structural characteristics of protein-protein interactions. H. Zhang et al. proposed a method by combining the high frequency modes of Gaussian network model and Gaussian Naive Bayes to identify hot spots, which are residues that contribute largely to protein-protein interaction energy. G. Hu et al. performed a comparative study of elastic network model and protein contact network for protein complexes in case of hemoglobin.Protein-protein interactions not only are limited to binary associations but also employ special topological structures to perform their biological functions. N. Bernabò et al. approached the comparison of two networks models obtained from two different text mining tools to suggest that actin dynamics affect spermatozoa postejaculatory life. J. Xu et al. also compared the topologies of network modes of drug-kinase interactions between established targets and those of clinical trial ones.Both protein structures and protein-protein interactions are involved in the subject of a growing number of pharmacological studies. F. Ye et al. identified that compounds DC_C11 and DC_C66 are two small inhibitors against protein-protein interactions between CARM1 and its substrates, while Y. Wang et al. identified that CID_70128824, CID_70127147, and CID_70126881 are three potential inhibitors for targeting the androgen receptor to treat prostate cancer. Finally, a review paper completes the issue. X. Li et al. introduced recent advances in the development of network models of identifying synergistic drug combinations.Altogether, we wish this issue has given a wider development of structural biology and systems biology with the advantage of biological network analysis as well as prospecting the future of this area towards translational bioinformatics and systems pharmacology.Guang HuLuisa Di PaolaFilippo PullaraZhongjie LiangIntawat Nookaew


Childhood adversity predicts adolescent suicidal ideation but there are few studiesexamining whether the risk of childhood adversity extends to suicidal ideation inmidlife. We hypothesized that childhood adversity predicts midlife suicidal ideation andthis is partially mediated by adolescent internalizing disorders, externalizingdisorders and adult exposure to life events and interpersonal difficulties.At 45 years, 9377 women and men from the UK 1958 British Birth Cohort Studyparticipated in a clinical survey. Childhood adversity was prospectively assessed at theages of 7, 11 and 16 years. Suicidal ideation at midlife was assessed by the depressiveideas subscale of the Revised Clinical Interview Schedule. Internalizing andexternalizing disorders were measured by the Rutter scales at 16 years. Life events,periods of unemployment, partnership separations and alcohol dependence were measuredthrough adulthood.Illness in the household, paternal absence, institutional care, parental divorce andretrospective reports of parental physical and sexual abuse predicted suicidal ideationat 45 years. Three or more childhood adversities were associated with suicidal ideationat 45 years .Psychological distress at 16 years partially mediated the associations of physical abuse, sexual abuse  with suicidalideation. Adult life events partially mediated the association of parental divorce  and physical  and sexual abuse with suicidal ideation at 45 years.Adversity in childhood predicts suicidal ideation in midlife, partially mediated byadolescent internalizing and externalizing disorders, adult life events andinterpersonal difficulties. Understanding the pathways from adversity to suicidalideation can inform suicide prevention and the targeting of preventiveinterventions. We hypothesized that: (1) childhoodadversity predicts suicidal ideation at midlife; and (2) the association of childhoodadversity and suicidal ideation is partially mediated through (n = 17 416). Analyses werebased on 9377 participants in a clinical survey at 45 years. The response rate for theclinical survey was 78% of those invited, representing 54% of the surviving population ;Sexual abuse by a parent .Childhood adversity was defined as exposure to traumatic events or chronic stressors  Study from 16 to 42 years wascalculated from the activity histories available from 1974 to 2009  .et al.n = 9377). A total of 30 datasets of the imputation were run andanalyses indicated that the measures were stable across the imputations. Parameterestimates from the 30 imputations were estimated using the MIM function in STATA.Imputed analyses are presented with unimputed prevalence figures. Multiple imputation, under the ‘missing at random’ assumption, was used to address theissue of missing data, using the ICE program in STATA. The measures described above wereincluded in the imputation equations. Employment status at 45 years, and social class at7 and 42 years were also included as they were significantly associated with attrition internalizing and externalizingdisorders at 16 years and (b) the adulthood life events andinterpersonal difficulties.Further logistic regression analyses assessed the associations of: (i) interpersonaldifficulties ; and (ii) adultdrinking problems (at 33 or 42 years) with midlife suicidal ideation, adjusting forgender, social class at 42 years and qualifications at 33 years. Finally, theassociations between childhood adversities and suicidal ideation were re-run,additionally adjusting for  internalizing disorders; (ii) externalizingdisorders; or (iii) interpersonal difficulties and problem drinking was investigatedusing the p < 0.05) and adjusted for gender where not significant(p ⩾ 0.05).All analyses tested for interactions between childhood adversity and gender: analyseswere stratified by gender if the interaction was significant.We carried out analyses to examine how much this adult adversity score might mediate theassociation between childhood adversity and suicidal ideation. The indirect percentage ofthe total mediation effect was between 3.28 and 8.63 for feeling that life was not worthliving and between 7.86 and 8.67 for depressive ideas. The percentage of mediationexplained was of little greater magnitude than the individual life events andinterpersonal difficulties (see Supplementary Table).The aims of our study were to assess whether the effects of childhood adversity on suicidalideation extended beyond adolescence to midlife and whether this was partially mediated byadolescent internalizing disorders, externalizing disorders and adult exposure to lifeevents and interpersonal difficulties. We confirmed that specific childhood adversitieswhich included illness in the household, paternal absence and divorce prospectively predictsuicidal ideation at 45 years even after adjustment for confounding and mediating factors.Retrospectively recalled parental sexual and physical abuse also show strong associationswith suicidal ideation at 45 years. Childhood adversity predicts adult life events,supporting continuity of exposure to adversity across the lifecourse. Adulthoodinterpersonal difficulties predicted suicidal ideation.et al.In terms of investigation of mediation, psychological ill-health in childhood partiallymediated the association of illness in the household, paternal absence and physical andsexual abuse. Adult life events partially mediated the association of paternal absence,divorce and physical and sexual abuse, while partnerships in adulthood and problem drinkingpartially mediated the association of sexual and physical abuse. We have confirmed ourhypotheses that some childhood adversities are associated with midlife suicidal ideation andthat this is partly mediated by subsequent life events, internalizing and externalizingdisorders, although the variance explained in mediation is small. Our findings supportKendler's aetiological framework for affective disorders (Kendler et al.et al.et al.et al.et al.et al.etal.et al.et al.Studies of youth suicide have demonstrated the importance of parent-related events in thefamily such as parental hospital admission with mental illness, parental divorce and maritaldisruption (Fergusson et al.et al.et al.et al.et al.et al.et al.et al.et al.et al.et al.et al.et al.In terms of other types of childhood adversity sexual (Fergusson et al.et al.et al. (et al.Lack of maternal and paternal care has been linked to adult suicidal ideation (Enns.et al.  state th. et al., althouget al.et al.et al.et al.The magnitude of the effects of the mediating factors of internalizing, externalizing andinterpersonal disorders may have been small due to methodological limitations: risk factorexposure misclassification, lack of key variables at different stages of the lifecourse andlack of coverage of all relevant risk factors within the three groups of mediators.Additionally we needed to dichotomize the mediating variables, rather than use continuousscores because of the skewed nature of their distribution. Thus we may have underestimatedthe effect of the mediators. Moreover, as the models are fully adjusted we only expected tosee small levels of mediation as the other covariates in the model also contribute to theoutcome. Looking at individual possible mediators on top of the effects of other covariatesmeans that individual mediators are unlikely to explain a large percentage of theassociations. However, in our study we may also be missing some key variables that areshaped by childhood adversity and transmit the risk of suicidal ideation across thelifecourse. One of these may be how the experience of adversity influences feelings ofself-worth, mastery and the ability to develop positive and trusting relationships. In turnthis may influence coping capacities and the ability to ask for help in a crisis (Gunnellet al.et al.et al.et al.et al.A common theme across studies of risk factors for suicidal ideation and suicide is lowsocial support and social isolation (Heikkinen et al.It is a strength that we had prospective measurements of childhood adversities andmediating factors at different life stages. To our knowledge this is the first study toexamine the role of mediators of prospectively measured childhood adversity and itsassociation with midlife suicidal ideation. It is a limitation that childhood sexual andphysical abuse were reported retrospectively (Colman et al.et al.et al.Suicidal ideation and completed suicide are not equivalent, and less than 1 in 200 of thosewith suicidal ideation proceed to suicide (Gunnell Some elements of childhood adversity have an impact on midlife suicidal ideation afteradjustment for mediating factors. This is in keeping with effects of adversity in criticalperiods in childhood having long-term consequences for mental health that may reflecteffects on the developing brain and neuroendocrine responses to stress.et al.Understanding the pathways from adversity to suicidal ideation can inform suicideprevention (Gunnell


The present special volume “Ecotoxicology in Tropical Regions” contains 19 research articles that were selected from presentations at two conference sessions at the SETAC Europe 24th and 25th meetings in Basel 2014 and in Barcelona 2015. The papers address major ecotoxicological issues in several tropical and sub-tropical countries including Brazil, Mexico, Costa Rica, Columbia, Nicaragua, Costa Rica, Ethiopia, Thailand, Vietnam, and Iran.In 2017, more than 16,000 scientists from 184 countries published a “Warning to humanity letter” . Similar results were reported in the paper by Sobrino-Figueroa Daphnia magna, commonly used to assess water quality. On the other hand, Khatikarn et al. In Moura et al. Macrobrachium and Cardina sp.) were more sensitive to pesticides than temperate crustaceans such as Daphnia magna. They discuss possible reasons for these differences and recommend using native species for first tier prospective ecological risk assessment (ERA).In the paper by Daam and Rico Crassostrea rhizophorae was evaluated for chemical biomonitoring. The authors found that this oyster, commonly associated with mangrove trees, is a suitable species for biomonitoring in Caribbean coastal systems. Six papers are dealing with pesticide environmental contamination in Costa Rica. Costa Rica has one of the richest biodiversity on Earth. It also has an extensive agriculture, with banana and pineapple as main export crops, and has the highest pesticide use per capita in Central America. Pesticide spray drift and runoff from plantations pose toxicity risks to aquatic organisms downstream these crops and to humans. The paper of Méndez et al. Caňo Negro wetland and watershed located in northern Costa Rica. Arias-Andrés et al. In the paper by Aguirre-Rubí et al. Three papers present studies from pesticide toxicity in Vietnam. Tam et al. A paper by Teklu et al. This Special Issue presents a unique collection of papers, describing various actual ecotoxicological issues in ten tropical and sub-tropical countries. As anthropogenic pressures and environmental problems that these and other tropical countries are currently facing will increase during the coming decades, it is essential that the scientific community carries out more research in tropical regions. We hope that the results and methodologies presented here will help policymakers to motivate stronger environmental protection measures to limit contaminant emissions and habitat destruction in tropical regions.Finally, we would like to thank all the reviewers who helped improve the quality of the research articles included in this Special Issue.