CesA gene family is limited. We have identified 22 CesA genes from bread wheat, which include homoeologs from each of the three genomes, and named them as TaCesAXA, TaCesAXB or TaCesAXD, where X denotes the gene number and the last suffix stands for the respective genome. Sequence analyses of the CESA proteins from wheat and their orthologs from barley, maize, rice, and several dicot species  revealed motifs unique to monocots  or dicots. Novel structural motifs CQIC and SVICEXWFA were identified, which distinguished the CESAs involved in the formation of primary and secondary cell wall (PCW and SCW) in all the species. We also identified several new motifs specific to monocots or dicots. The conserved motifs identified in this study possibly play functional roles specific to PCW or SCW formation. The new insights from this study advance our knowledge about the structure, function and evolution of the CesA family in plants in general and wheat in particular. This information will be useful in improving culm strength to reduce lodging or alter wall composition to improve biofuel production.Cellulose is the primary determinant of mechanical strength in plant tissues. Late-season lodging is inversely related to the amount of cellulose in a unit length of the stem. Wheat is the most widely grown of all the crops globally, yet information on its  Maize and rice possess 13 and 11 CesA genes, respectively [HvCesA1, HvCesA2, and HvCesA6 make PCW, and HvCesA4, HvCesA7, and HvCesA8 form SCW [HvCesA3, 5 and 9 are different from both the groups because of their unique tissue-specific transcript levels [CesA gene family were spread across the genome although some genes were clustered together [Cellulose is the primary determinant of mechanical strength in plants , 2. It i subunit . Arabidox groups –10. Mutaormation , 12. The AtCesA6 , 14. AtCectively , 16. Barform SCW . HvCesA3t levels . Mappingtogether .Plant CESAs belong to family 2 of glycosyl transferases (GT2), which catalyze beta linkage between the glycosyl residues. CESAs are intrinsic plasma membrane proteins with their catalytic domains extending into the cytoplasm . Each ofA CESA protein of higher plants possesses eight transmembrane domains (TMDs), which are believed to form a pore in the plasma membrane to allow extrusion of the newly synthesised glucan chain. Two zinc-finger domains (ZnF), which are highly homologous to the RING-finger motif, are present on the cytoplasmic face close to the amino terminus . The cenRhodobacter sphaeroides demonstrated the involvement of a single catalytic site in the formation of beta-1,4-glycosidic bond of the glucan chain [Motifs are the conserved groups of residues in proteins, which can be associated with structural and functional variability across species. A highly conserved motif, CXXC, which is located within the ZnF, distnguishes CESAs from the CSL (cellulose synthase-like) proteins , 10. Cryan chain . Computaan chain , 26–28. CesA gene family has not yet been compiled from wheat, the most widely grown crop in global agriculture. Functional classification of the CesA genes in cereal crops has proved helpful in associating various genes with culm or stalk strength [CesA gene family from wheat. To understand the involvement of the different CesAs in primary or secondary wall formation in grasses or dicot plants, we have identified unique sequence motifs. Sequence comparisons of the PCW and SCW TaCesA genes were performed at both the DNA and protein levels. Phases of intron evolution were predicted and compared between the groups of the TaCesA genes involved in the formation of PCW or SCW. Unique motifs were identified among the representative monocot and dicot species. RNA-seq expression profiling of the TaCesA genes revealed unique, homoeolog-specific expression patterns in different tissues.The strength , 17. In http://plants.ensembl.org/Triticum_aestivum/Info/Index) generated by the International Wheat Genome Sequencing Consortium (IWGSC) [http://webblast.ipk-gatersleben.de/barley/) made it possible to isolate full-length barley CesA sequences [Triticum urartu and Aegilops tauschii, A and D genome progenitors of wheat, respectively, were also explored to identify full-length CesA genes for the sequences missing in hexaploid wheat. The homoeologs were first identified from Ensembl Plant database followed by amino acid sequence alignment for the presence of conserved motifs and domains. Highly variable class-specific regions (C-SRs) present in different CesAs were used to differentiate the homoeologous genes from each other (The conserved cellulose synthase domains from barley CESA proteins was used as query to perform tBLASTn search with Chromosome Survey Sequence (CSS) ( (IWGSC) . Availabequences . Genome ch other .CesA genes were identified through alignment of the wheat CesAs with those from Arabidopsis, barley, rice and maize. The ortholog of each gene was selected based on the sequence identity and query coverage, presence of all domains and motifs similar to the query sequence, Amino acid content/size and distance among various new motifs identified in this study relative to the query sequence. Arabidopsis, rice and maize CesA sequences were retrieved from Phytozome v9.1: Home (http://www.phytozome.net/) [Orthologs of various me.net/) .TaCesA genes, comparative studies for gene structure were performed only for the genes that were specific for PCW and SCW cellulose synthesis. Based on analysis of the orthologs, TaCesA4, 7 and 8 were characterized as one-to-one orthologs of SCW-specific, TaCesA1, 2, and 6 as PCW-specific, and TaCesA3, 5 and 9 as partially redundant to the PCW CesAs. The homoeologous copies of each gene shared 95–99% sequence identity in addition to all the motifs and domains. Therefore only one copy among the three homoeologs was used for comparative analysis. Intron-exon boundaries and translation start and stop sites were predicted through alignments of full-length genomic copies of TaCesA genes with their corresponding cDNA sequences. The introns and exons were drawn to scale for all the genes as indicated by the cDNA-genomic sequence comparisons. Phases of intron evolution were predicted using Plant Intron Exon Comparison and Evolution database (PIECE) (http://wheat.pw.usda.gov/piece/) [Although in this study we identified 22 /piece/) .http://www.genome.jp/tools/clustalw/). Color Align Conservation tool  was used to differentiate the conserved patterns of aligned sequences. Conserved domains and motifs were identified by manual search in the aligned sequences.Amino acid sequence similarity of TaCESA protein sequences was determined by multiple sequence alignment , Beta vulgaris (BvCESA), Eucalyptus grandis (EgCESA), Glycine max (GmCESA), Gossypium hirsutum (GhCESA), Hordeum vulgare (HvCESA), Oryza sativa (OsCESA), Populus trichocarpa (PtCESA), Solanum tuberosum (StCESA), Zea mays (ZmCESA) were retrieved from NCBI (http://www.ncbi.nlm.nih.gov) [22 newly identified wheat CESA proteins were used to deduce their phylogenetic relationships. Protein sequences for nih.gov) . An unronih.gov) , 34. Evonih.gov) . All pos-42g29300; OsCESA11, LOC_OS06g39970; HvCESA1, AY483150; HvCESA2, AY483152; HvCESA3, AY483151; HvCESA4, AY483154; HvCESA5/7, AY483153; HvCESA6, AY483155; HvCESA8, AY483156; HvCESA9, AK367031; PtCESA6, XP_002319002; EgCESA5, XP_010063196; StCESA3, XP_006354075; GmCESA2, XP_003531396; GhCESA5, AFB18634 and BvCESA2, XP_010678670.GenBank accession numbers for CESA amino acid sequences used to generate the phylogenetic tree are: AtCESA1, AF027172; AtCESA2, AF027173; AtCESA3, AF027174; AtCESA4, AB006703; AtCESA5, AB016893; AtCESA6, AF062485; AtCESA7, AF088917; AtCESA8, AL035526; AtCESA9, AC007019; AtCESA10, At2G25540; ZmCESA1, AF200525; ZmCESA2, AF200526; ZmCESA3, NP_001292792.1; ZmCESA4, AF200528; ZmCESA5, AF200529; ZmCESA6, AF200530; ZmCESA7, AF200531; ZmCESA8, AF200532; ZmCESA9, AF200533; ZmCESA10, AY372244; ZmCESA11, AY372245; ZmCESA12, AY372246; ZmCESA13, KJ874174; OsCESA1, AF030052; OsCESA2, D48636, OsCESA3, BAD30574; OsCESA4, AK100475; OsCESA5, BAD30574; OsCESA6, XM_477282; OsCESA7, XM_477282; OsCESA8, XM_477093; OsCESA9, XM_477093; OsCESA10, LOC_OCesA genes was performed using publicly available RNA-seq data from two different databases (http://wheat-urgi.versailles.inra.fr/Seq-Repository/RNA-Seq) at McGill University and Genome Quebec Innovation Center. First dataset was a non-oriented library with five wheat organs analysed in duplicates at three development stages for each of the organs. The five organs taken into consideration with respect to developmental stages were root , leaf , stem , spike  and grain . The second dataset was the oriented library with five wheat organs  with five conditions pooled for 4 lines per organ [Gene expression profiling for 21 of the wheat er organ .The abundance of transcripts from RNA-Seq data was reported using the estimated counts quantified by a programme Kallisto (v0.42.1) . Counts-http://plants.ensembl.org/Triticum_ aestivum/Info/Index) generated by the International Wheat Genome Sequencing Consortium to identify the orthologs of various CesA genes from bread wheat corresponding to the barley CesA sequences [TaCesA genes were isolated, six of which were partial  that differentiated them from each other. The wheat orthologs of the CESA proteins of other species exhibited a similarity of 70–80% at the amino acid level with Arabidopsis and 90–95% with rice and barley. The TaCesA genes ranged from 4044 to 5251 bp in length and contained 9–13 introns. The ensembl IDs of all the newly identified wheat CesA genes are given in We queried the Chromosome Survey Sequence (CSS) . As expected the chromosomal locations of different CesA genes followed the trend reported earlier in the syntenic species barley [TaCesA4A, B, and D mapped in respective genomes to chromosome 1; TaCesA7B and D to chromosome 3; and TaCesA8B and D to chromosome 5. Similarly, the homoeologs of TaCesA1, 2, 3, 5, and 6 mapped to chromosomes 2, 5, 5, 1 and 6 of the respective genomes. However, TaCesa9B mapped to chromosome 2B, unlike its ortholog from barley, which is located on chromosome 6. The approximate location of TaCesA genes and their homoeologs is presented in The newly identified wheat s barley . TaCesA4TaCesA6, was 5251 base pairs (bp) and the shortest, TaCesA4, was 3923 bp in length. The size variations among different CesA genes arose mainly from the number and length of introns  were similar in length to those of the SCW forming genes (2.9–3.2kb). Exon-intron boundaries were random in all the genes studied, which was in contrast to the conserved boundaries reported in other species [The introns in PCW  species . The PCWTaCesA genes, all of the introns had identical phase distributions: introns 1, 3, 7, 8, 9, 10, 12, and 13 occurred in 0 phase, introns 2, 4, and 11 were in phase 1, and introns 5 and 6 occurred in phase 2. In contract, SCW TaCesA genes exhibited variable patterns of intron phase distribution. Introns 2, 5, 6, and 7 in CesA4 had 0 phase distribution, introns 1 and 3 had 1, and intron 4 had a phase distribution of 2. TaCesA7 also had introns with all three types of phase distribution; introns 2, 6, 7, 8, 9, 11, 12 were in phase 0, introns 1, 3, and 10 in phase 1, and introns 4 and 5 in phase 2. CesA8 similarly had introns 1, 4, 5, 6, 8, and 9 in phase 0, introns 2 and 7 in phase 1, and intron 3 in phase 2 , respectively. To identify group-specific changes in primary and secondary cell wall CESA proteins, AA sequences from all TaCESAs were aligned. All the complete CESA proteins possessed the already known, specific CESA domains, such as a ZnF (CX2-CX14-ACX2-CX4-CX2-CX7-GX3-CX2-C) near the N-terminus of the derived amino acid sequence . All theMajor differences among TaCESAs resulted from the deletion of up to 45 AAs in hypervariable regions. The N-terminii of the PCW TaCESAs possessed more highly conserved motifs and fewer deletions in comparison to the SCW TaCESAs. ZnF consisted of 46 AAs in the predicted TaCESAs, with the exception of an 8 AAs deletion in TaCESA7 and its homoeologs, resulting in the following domain: CX2-CX6-ACX2-CX4-CX2-CX7-GX3-CX2-C as compared to the known domain (CX2-CX12-FXACX2-CX2PXCX2-CXEX5-GX3-CX2C), where X is any amino acid . Four ofA new motif distinguishing the PCW CESAs from the SCW CESAs was found within the ZnF. The motif, CQIC, was identified within the small motif, CXXC, reported earlier for differentiating CESAs from the CSL genes . This moThe two motifs, CQIC and SVICEXWFA, distinguished the PCW from the SCW CESAs . That thMotif analysis was performed by aligning CESA proteins from Arabidopsis, barley, maize, rice and wheat. Arabidopsis CESA4 and its orthologs from wheat, barley, maize, and rice exhibited 73–74% sequence similarity. In the case of SCW, nine motifs ranging from 2–15 amino acid residues in length provided ortholog-specific identity to the SCW CESAs from different species . These mTwo PCW CESAs, AtCESA1, 3 and their orthologs from other species differed from AtCESA6 and its orthologs in structural features. AtCESA1 and 3 were highly similar (77–79%) to the corresponding orthologs from barley, maize, rice and wheat. Four motifs in TaCESA6 and three in TaCESA1 orthologs differentiated them from each other and all other CESAs .Arabidopsis thaliana (AtCESA6), Beta vulgaris (BvCESA2), Eucalyptus grandis (EgCESA5), Glycine max (GmCESA2), Gossypium hirsutum (GhCESA5), Populus trichocarpa (PtCESA6) and Solanum tuberosum (StCESA3). The CESA2 and its orthologs from grasses were compared with its orthologs from dicot species. For this particular gene nine motifs were highly conserved in the orthologs from grasses  but not in Arabidopsis. To confirm the specificity of these motifs for grasses, we retrieved the sequences of TaCESA2 orthologs from seven dicot species:  grasses . But in CesA genes had occurred prior to the separation of monocots and dicots.The evolutionary history of the CESAs was inferred from the analysis involving 70 CESA protein sequences from different species. An unrooted phylogenetic tree revealed that the orthologs from Arabidopsis, barley, beet, cotton, maize, poplar, potato, rice, rose gum, soybean and wheat were grouped together. Branch lengths, which are indicative of the evolutionary distances were used to interpret the phylogenetic tree . The parTaCesA genes was studied in five organs at three development stages. We left out the TaCesA9 gene because it was represented by a highly truncated cDNA. A heat map displaying transcript abundance of the CesA genes from different wheat tissues is shown in Gene expression of 21 of the 22 TaCesA4A, B, D, TaCesA7B, D and TaCesA8B, D genes, all involved in SCW synthesis. These genes were highly expressed in the mature tissues, for example, stem collected soon after anthesis, and at very low levels in the PCW formation , whereas TaCesA1D was expressed at higher level. The transcript abundance of TaCesA1A was highest in the grain tissues at 2DAAs whereas TaCesA6B homoeolog was moderately expressed. The expression level of TaCesA1B was moderate in the root and grain tissues.Group II comprised the PCW ng cells . For exaCesA is a multigene family consisting of more than eight members in higher plants [CesA genes in wheat remains undocumented. Most studies about structural and functional characterization of CesAs have been performed in Arabidopsis [CesA gene family consists of nine genes , Chlorophytes , and pteridophyte (Selaginella moellendorffii).A similar trend of intron phase distribution and motif conservation was observed when we compared CESA1 of We also identified new, highly conserved motifs among the CESA orthologs of five species . Despite the variable protein sequence of each member of the CESA family among the orthologs from various species, the organization of the motifs remained conserved.TaCesA4, 7, 8) and their homoeologs were co-expressed in the mature stem tissues  and their homoeologs from the A, B and D genomes of wheat constituted a second group involved in PCW synthesis.RNA-seq expression profiling revealed that the three SCW genes  formation in higher plants. Further characterization of the motifs would be needed, however, to establish their respective biological roles. Several new motifs identified in this study would be useful as signatures for the identification of orthologs of the CesA genes from various species. The compilation of the CesA gene family in bread wheat along with the expression patterns and genomic map positions of individual members will be helpful in improving culm strength for reduced lodging as well as improving the straw for biofuels.We have identified 22 S1 Text(DOCX)Click here for additional data file.S2 Text(DOCX)Click here for additional data file.


S-(2-succino)cysteine (2SC). Previous studies have demonstrated that succination of proteins including glyceraldehyde 3-phosphate dehydrogenase (GAPDH), kelch-like ECH-associated protein 1 (KEAP1) and mitochondrial aconitase (ACO2) can have profound effects on cellular metabolism. Furthermore, immunostaining for 2SC is a sensitive and specific biomarker for HLRCC tumours. Here, we performed a proteomic screen on an FH-mutant tumour and two HLRCC-derived cancer cell lines and identified 60 proteins where one or more cysteine residues were succinated; 10 of which were succinated at cysteine residues either predicted, or experimentally proven, to be functionally significant. Bioinformatic enrichment analyses identified most succinated targets to be involved in redox signaling. To our knowledge, this is the first proteomic-based succination screen performed in human tumours and cancer-derived cells and has identified novel 2SC targets that may be relevant to the pathogenesis of HLRCC.Inherited mutations in the Krebs cycle enzyme fumarate hydratase (FH) predispose to hereditary leiomyomatosis and renal cell cancer (HLRCC). Loss of FH activity in HLRCC tumours causes accumulation of the Krebs cycle intermediate fumarate to high levels, which may act as an oncometabolite through various, but not necessarily mutually exclusive, mechanisms. One such mechanism, succination, is an irreversible non-enzymatic modification of cysteine residues by fumarate, to form  Mutatioectively ,5,6,7,8.patients ,10,11,12patients , althougpatients ,14,15,16In order to verify the presence of succinated proteins in the FH-mutant tumour-derived cell lines UOK262  and NCC-As previously reported, 2SC modification is very specific to cells with elevated fumarate ,21,22 an2SC targets identified an HLRCC tumour and tumour-derived cell lines. Succinated proteins are listed alphabetically by gene symbol, with succinated cysteine residues indicated. Targets highlighted in red indicate cysteine residues with either predicted or experimentally proven roles in protein function . T = HLRS-nitrosylating activity of thioredoxin [S-transferase 3 (MGST3) is succinated at Cys-150 and Cys-151, both of which are also targets of S-palmitoylation [Hierarchical clustering and gene ontology analyses suggest that most of the succination targets identified are involved in redox homeostasis or subject to ROS regulation . This isoredoxin . Isoformoredoxin . Succinaoredoxin . Furtheroylation . Site-diCell lines were cultured as previously described . The UOKTumour and normal kidney tissue and cells were homogenized and sonicated in Urea-SDS buffer and protein extracts separated by SDS-PAGE and processed for trypsin digestion and MS/MS analyses as previously described . DatabasAnalyses of tumours and cell lines by IHC and IB were performed as previously described ,46. The SNCAsynuclein, alpha (non A4 component of amyloid precursor)ARandrogen receptorPARK2Parkinson disease  2, parkinSOD1superoxide dismutase 1, solublePIAS2protein inhibitor of activated STAT, 2PTENphosphatase and tensin homolog; Tumour suppressorMTA1metastasis associated 1AKT1v-akt murine thymoma viral oncogene homolog 1MAP3K5mitogen-activated protein kinase kinase kinase 5NDUFA8NADH dehydrogenase (ubiquinone) 1 alpha subcomplex, 8Functional classification of succinated proteins was performed using DAVID Bioinformatics Resources v6.7 , NIH, Frederick, MD, USA) ,42. ProtRecent technological advances in mass spectrometry have facilitated high throughput screening of post-translational modifications in proteins. Here, we analysed cell lines and a tumour derived from HLRCC patients to try to identify the scope of the succinated proteome under FH-deficient settings. Succination of cysteine residues is a candidate mechanism for FH-associated oncogenesis, which is corroborated by genetic and biochemical evidence generated from the analyses of Fh1 knockout mice and gene expression profiling and exome sequencing of type 2 RCC ,26,61. I


Lysinibacillus sphaericus CBAM5, was isolated from subsurface soil of oil well explorations in the Easter Planes of Colombia. This strain has potential in bioremediation of heavy-metal polluted environments and biological control of Culex quinquefasciatus. According to the phylogenetic analysis of 16S rRNA gene sequences, the strain CBAM5 was assigned to the Lysinibacillus sphaericus taxonomic group 1 that comprises mosquito pathogenic strains. After a combination assembly-integration, alignment and gap-filling steps, we propose a 4,610,292 bp chromosomal scaffold. The whole genome  revealed strong functional and syntenial similarities to the L. sphaericus C3-41 genome. Mosquitocidal (Mtx), binary (Bin) toxins, cereolysin O, and heavy metal resistance clusters from nik, ars, czc, mnt, ter, cop, cad, and znu operons were identified. Lysinibacillus sphaericus is one of the bacteria used as a bio-insecticide as part of vector control programs against tropical diseases, such as malaria, filariasis, yellow fever, dengue fever and West Nile virus  analysis. In the contig 19 (EWH31640:EWH31645) we found a typical cluster consisting of dnaA, dnaN, recF, gyrA and gyrB boxes. The predicted genes dnaB, dnaI, dnaG, dnaE, holA, holB, priAB, polA and recA were also found spread in the chromosomal and extrachosmomal sequences. The replication termination site of the chromosomal scaffold is believed to be localized near 2.92 Mbp in the contig 14. According to GC skew analysis, the coding bias for the two strands of the chromosome is for the majority of CDSs to be on the outer strand from 0 to ~2.92 Mbp, and on the inner strand from ~2.92 Mbp to the end of the chromosomal scaffold  and parE (EWH32538), which encode the subunits of the chromosome-partitioning enzyme topoisomerase IV. Similar to previous reports [rtp (replication terminator protein-encoding gene) in the chromosomal assembly of CBAM5.The origin of replication of the chromosome of  reports ,51, we dLysinibacillus sphaericus CBAM5 displays 28 CDSs annotated as transposases, including three allocated in the extrachromosomal sequences. The most frequent families are IS1182, IS3 and IS4. In addition, four incomplete prophage regions were identified as follows: Thermus phage φOH2 (contig 12), Burkholderia phage ST79 (contig 14), and two regions comprising the Clostridium phage φSM101 (contigs 14 and 28). Prophage regions φOH2 and ST79 included putative encoding sequences for tail, lysis and baseplate proteins. None of the reported phages has been described in the Colombian strain L. sphaericus OT4b.31 [ OT4b.31 .L. sphaericus CBAM5 shows a wide repertoire of potential encoding sequences in terms of mosquitocidal toxins. In the contig 11, we found Mtx1 (EWH35097) and Mtx2 (EWH35034) CDSs located in an identical cluster as Hu et al. [L. sphaericus C3-41. This cluster includes two insertion sequences, one of them consisting of a disrupted transposase between the mtx1 and mtx2, as well. One Mtx3 CDSs (EWH32377) was found in contig 14. Upstream of this sequence, we could identify some IS3 family mobile elements and putative DeoR family transcriptional regulators. In addition, we found one hypothetical toxin from the family Mtx2 (PFam PF03318) in contig 11 (EWH35106) and a putative cereolysin O CDS (EWH31995) being described to be active against the German cockroach Blattela germanica[The genome of u et al.  describegermanica in contibinA (EWH32662) and binB (EWH32663) of L. sphaericus CBAM5, which are the main source of its larvicidal activity [L. sphaericus C3-41 , two CDSs for phage integrases in the 5’ start of the 35-kb fragment. L. sphaericus CBAM5 also share a germination gene cluster equivalent to the B. anthracis plasmid pXO1 and the BinA/BinB cluster of L. sphaericus C3-41, having a GerXB-KA-XC gene cluster upstream of a transposase [binA-binB genes across the sequences of L. sphaericus CBAM5, C3-41 and 2297, we found an equivalent homology of putative transposases with different length and disruption points. The strain CBAM5 has two mobile elements of 459 and 312 bp in length, which is similar to strains 2297 and CBAM5 showing a probable transposase pseudogene with 1,110 bp and 591 bp in length, respectively  being the remnant of a potential phage infection. Even though we cannot confirm the presence of additional BinA-B CDS sequences in the genome of L. sphaericus CBAM5, we suggest further research to confirm the participation of phage infections on the evolution of larvicidal toxins in the strain CBAM5.The binary toxin genes activity , were fonsposase ,53. CompL. sphaericus CBAM5 exhibits 21 CDSs described as surface (S) layer proteins or S-layer homologs in its genome. The fragment covered from EWH35069 to EWH35072 includes four CDSs encoding for a variable protein, a putative S-layer associated protein, a P60 invasion-associated protein and a N-acetylmuramoyl-L-alanine amidase. Probably the genes located in this fragment may participate in the larvicidal activity of the strain CBAM5, given that the same genes have been described as differentially expressed in virulent infections of Lysteria monocytogenes[slpC gene previously described in native strains [cytogenes. A totalarsRBCDA, arsRBC, cadCA, mntABCD, nikABCDE, terD-terD-terD, zurR-znuBC, and czrA-czcD-csoR-copZA. We could identify various genes probably involved in metal(loid) resistances spread across the genome  [A total of 64 CDSs corresponded to encoding sequences involved in responses against toxic metal(oid)s. Among those coding sequences, we found the following operons: t Cr(VI) . Howevert Cr(VI) ,56.L. sphaericus CBAM5 in polluted environments, and supported by the identification of genes in Additional file L. sphaericus CBAM5 may have acquired the arsC gene through recent events of horizontal gene transfer as a possible adaptation to polluted environments. However, we found highly similar homologues of heavy metal resistance proteins of the CBAM5 strain in microorganisms isolated from non-polluted environments . FurtheLysinibacillus sphaericus CBAM5 was isolated from drilling mineral base oil samples at the subsurface soil level. By comparing the chromosomal sequences between L. sphaericus strains CBAM5 and C3-41, we identified distinctive similarities of the DNA homology group IIA. The evidence of the binary toxins allocated in a conserved cluster delimited by mobile elements, resembles a probable phage invasion in the DNA subgroup IIA of the Lysinibacillus sphaericus species. Along with the biological control potential given by the Mtx, Bin and cerolysin toxins, L. sphaericus CBAM5 displays encoding sequences for S-layer proteins and heavy-metal efflux pumps, which may confer resistance to As, Sb, Ni, Zn, Cu, Cd, Te, Cr, Mn and Co in polluted environments.The authors declare that they have no competing interests.LL performed the DNA and sequencing experiments. TDP performed antibiotics, microscopy and bioinformatics analysis. All authors drafted, read and approved the final manuscript.Associated record according to the MIGS recommendations.Click here for fileLysinibacillus sphaericus CBAM5 growth in acetate broth. (A) Gram staining of vegetative cells after 6 hours of growth. (B) Schaeffer-Fulton staining of sporulating culture after 24 hours of growth.Light microscopy of Click here for fileLysinibacillus sphaericus CBAM5. The micrograph was obtained on a JEOL JSM-5800LV (Japan) scanning electron microscope at an operating voltage of 20 kV and 10000× magnifications.Scanning electron micrograph of Click here for fileLysinibacillus sphaericus CBAM5.Genes possibly involved in metal(loid) resistances identified in the genome sequence of Click here for file


Auxin is a good candidate for one of these Ca2+-activating signals. However, the role of auxin-induced Ca2+ signaling is poorly understood. Here, we will provide an overview of possible developmental and physiological roles, as well as mechanisms underlying the interconnection of Ca2+ and auxin signaling.Due to their sessile lifestyles, plants need to deal with the limitations and stresses imposed by the changing environment. Plants cope with these by a remarkable developmental flexibility, which is embedded in their strategy to survive. Plants can adjust their size, shape and number of organs, bend according to gravity and light, and regenerate tissues that were damaged, utilizing a coordinating, intercellular signal, the plant hormone, auxin. Another versatile signal is the cation, Ca How do you survive when you are sessile? The answer is that you need to be extremely flexible and even change your body plan according to actual limitations imposed by the environment. Herein lays the key to the success of plants. They can develop highly complex and elaborate body plans under optimal conditions, while less favorable environments results in much smaller plants with reduced body plan complexity. The ability to adjust the number and size of organs, regenerate damaged or destroyed organs and to (re)orient growth according to light and gravity are a few examples of the mechanisms that illustrate plants’ flexible, adaptive growth. etc.) impact on auxin’s action, thereby optimizing the plant’s growth and development [At the very core of this developmental flexibility is the plant hormone auxin ,2,3. Sevelopment . 2+, is one of the most universal second messengers; a signal that relays a primary signal, such as derived from an activated receptor, from the surface of the cell to intracellular targets. It can be found in ancient bacteria and throughout eukaryotic lineages, where it is essential to support life [2+ in cellular signaling processes is roughly summarized in the famous quote from Otto Loewi in 1959: “Ja Kalzium, das ist alles!” . Furthermore, in plants, Ca2+ is a fundamentally important second messenger, as demonstrated by its involvement in a multitude of essential cellular process, ranging from cell division, cell growth/shrinkage, secretion, transcriptional regulation, cellular polarity, etc., by which it impacts on stomatal aperture regulation, responses to light, responses to biotic and abiotic stresses, immunity a and responses to multiple plant hormones, including response to auxin [2+ signals can be regulated by so many different cues and have such a broad impact on cellular processes, it is not unlikely that Ca2+ acts to integrate multiple cues in a single output. Here, we will focus on the effects of auxin on Ca2+ and vice versa. The divalent cation, Caort life ,46. The to auxin ,48,49. B2+ sensitive dyes and, recently, also genetically encoded Ca2+ sensors, strong correlations could be observed between Ca2+ and auxin signaling. This is most apparent after exogenous application of synthetic and natural auxins, which induce a rapid, transient increase in cytosolic Ca2+ concentration in wheat leaf protoplasts [Arabidopsis roots [2+ concentration was observed to occur within minutes after auxin application, demonstrating that auxin is a potent inducer of Ca2+ signals. One of the important questions that remain to be answered is how these auxin-induced Ca2+ signals are generated. Generally, rapid and local Ca2+ signals generated in the cytosol depend on a Ca2+ current that is driven by a steep concentration gradient between the cytosol (nanomolar range) and its adjacent organelles or extracellular space (millimolar range). This allows the bringing about of a rapid and strong Ca2+ rise in the cytosol required for immediate activation of an appropriate response by the simple opening of a few Ca2+ channels [Via Catoplasts ,51, maiztoplasts ,53,54, pis roots  and closis roots . In nearchannels ,46,56.2+. The apoplast is an important source for auxin-induced Ca2+ signals. These could be prevented by application of membrane impermeable Ca2+ chelators and Ca2+ channel inhibitors [2+ changes in leaf wheat protoplasts and root hairs depend strongly on the Ca2+ concentration in the extracellular medium [+ and Ca2+. From these measurements, it was inferred that auxin activates cation channels in the plasma membrane that facilitate Ca2+ influx and K+ efflux. Consistently, both auxin-induced currents could be inhibited by nifedipine and verapamil [2+ channel blockers that were reported to inhibit outward rectifying K+ channels [2+ channels to generate auxin-induced Ca2+ signals. Importantly, a large portion of wheat leaf protoplasts showed a LiCl-sensitive, biphasic Ca2+ signal after auxin treatment, suggesting the involvement of intracellular Ca2+ stores [2+ stores contribute to auxin-induced Ca2+ signals. Unfortunately, most of the used dyes and genetically encoded Ca2+ sensors did not yield the needed resolution to detect intracellular sources for auxin-induced Ca2+. Recently, the genetically encoded Ca2+ sensor, aequorin, was targeted to the Golgi apparatus and revealed that application of the synthetic auxin analogue, 2,4-D , caused a steady/slow decrease of its Ca2+ content, suggestive of a passive Ca2+ loss [2+ fluxes, one would require using a broad range of subcellularly targeted Ca2+ sensors that were recently developed [In plants, the apoplast, the vacuole, the endoplasmic reticulum and all smaller organelles could serve as sources for auxin-induced Cahibitors ,57, and r medium ,51,58. Mr medium ,60. In terapamil ,60, two channels . Togethea2+ loss . To be aeveloped ,64,65,66TIR1/AFB E3-ligase, Aux/indole-3-acetic acids (Aux/IAAs) and auxin response factors (ARFs)  . The Aux cytosol ,54,67,68bidopsis . Interes mutants , suggestor, La3+ . This to2+ changes. An alternative auxin signaling pathway based on the stabilized interaction between F-box protein SKP2A and cell cycle transcription factor DPB was recently proposed to explain the effects of auxin on cell cycle progression . However+ and Cl− across the plasma membrane [2+ signaling. However, available data are indirect and inconclusive as exemplified in auxin-induced stomatal opening. Firstly, it is well established that auxin induces stomatal opening [2+ signals and cytosolic acidification in guard cells [2+ via ethylene glycol tetraacetic acid (EGTA) prevented auxin-induced stomatal opening [2+ is required for auxin-induced stomatal opening. Thirdly, activation of apoplastic ABP1 via exogenous application of a specific antibody induced stomatal opening, while exogenous application of polyclonal antibodies could interfere with auxin-induced stomatal opening [2+ during auxin-induced stomatal opening. However, in mutants defective in AUX1-mediated IAA uptake, IAA could no longer counteract abscisic acid (ABA)-induced stomatal closure [2+-dependent portion of auxin-induced protoplast swelling was suggested to be independent of apoplastic ABP1 [A second main auxin receptor is defined by Auxin Binding Protein 1 (ABP1) ,70,71,72membrane ,77 and rmembrane ,78,79,80 opening , which hrd cells . Secondl opening  indicati opening . Togethe closure , arguingtic ABP1 . TIR1/AFB signaling cascades are interconnected: in conditional knockdown lines for ABP1, SCFTIR1/AFB-regulated transcripts were less auxin-sensitive [2+ acts as a coordinating signal between both signaling cascades  is the phosphorylation-dependent interaction with a 14-3-3 protein. This interaction, and, thus, proton extrusion, depends mainly on phosphorylation of the penultimate Thr in its C-terminus [C-terminus by the protein kinase PKS5/CIPK11, which acts in concert with the Ca2+ binding protein, ScaBP1/CBL2 [2+-dependent root growth inhibition in high auxin levels [2+-dependent inhibition could be part of a feedback mechanism that keeps apoplast acidification in check, as arabinogalactan glycoproteins (AGBs) are proposed to act as pH-sensitive Ca2+ sources in the periplasm [2+ concentration that can contribute to cytosolic Ca2+ signals that inhibit AHA activity  family, together with CBL interacting kinases (CIPK) or Ca2+-dependent kinases (CPKs), mediate Ca2+-dependent phosphorylation of K+ channels to fine-tune their activities. Furthermore, during auxin’s effect on osmoregulation, Ca2+ could be an important signal, as high extracellular Ca2+ impairs acid-induced growth, which is associated with reduced inward K+ currents [+ transporters, TRH1/AtKT3/AtKUP4 [+ transporter AKT2 [2+ on K+ channels could be related to regulating osmotic pressure for both cell elongation and for auxin transport. Besides a possible modulation of Hin roots ,112,113.currents . Moreove3/AtKUP4 ,115 and 3/AtKUP4 , and CIP3/AtKUP4  (which cter AKT2 ) have be2+ availability [2+ in auxin transport. As early as the nineteenth century, Charles Darwin already recognized the existence of a mobile signal that moves between the site of light perception and the site of elongation growth during phototropic bending . Ever silability ,124,125.The minimal mechanistic constituents of polar auxin transport have been delineated in the chemiosmotic polar diffusion hypothesis ,128,129 +/IAA− symporters [2+ was found to have a profound, but indirect, impact on AUX1-mediated H+/IAA− symport, via effects on plasma membrane H+ ATPase activity [2+/H+ exchangers, CAX1 and CAX3, were insensitive to IAA’s inhibitory effect on ABA-induced stomatal closure. This defect in IAA sensitivity could be fully rescued by the lowering of apoplastic pH in the mutants. This illustrates how vacuolar Ca2+ homeostasis processes can impact on auxin uptake. Similarly, cax1 mutants show reduced lateral root densities and show IAA-resistant root growth [aux1 mutant phenotypes [Since the formulation of this model, most of the underlying molecular machinery has been identified and characterized in detail. Besides passive auxin entry, auxin was found to be actively taken up into the cell . This ismporters . They armporters ,135, latmporters  and phylmporters . Interesactivity   and TOUCH 3 (TCH3), respectively [2+ can have a positive, as well as a negative, impact on PID activity and, thus, on ABCB-mediated auxin transport. Conversely, as D6PKs are alsoAGC-type kinases, it is tempting to speculate that these kinases could also be regulated by Ca2+-binding proteins to regulate PIN-mediated auxin transport activity. The other rate-limiting aspect of the polar auxin transport is auxin efflux c. This pectively . This su2+ could impact on auxin transport rates via direct effects on the auxin transport machinery. However, there are probably even more mechanisms by which Ca2+ can impact on polar auxin transport. One example of this is the recent finding that overexpression of SAUR19, a member of an early auxin responsive protein family that binds with high affinity to calmodulin [These examples demonstrate how Calmodulin , promotelmodulin .2+ impacts on the speed of auxin transport, it was already proposed in 1984 that reduced polar auxin transport under low Ca2+ conditions was the result of both a lower velocity and a lower capacity for auxin transport [2+ controls not only the activity, but also the relative abundance of auxin transporters at the plasma membrane. While the above examples demonstrate how Caransport . This imtrans-Golgi Network (TGN) to the plasma membrane. Via clathrin-mediated endocytosis, PIN proteins are removed from the plasma membrane to early endosomes/TGN from which they can be targeted to the vacuole for degradation or recycled for exocytosis at the PM [2+ on PIN abundance at the plasma membrane is determined by the balance between exocytosis and endocytosis.Newly biosynthesized PINs are trafficked from the endoplasmic reticulum, via the Golgi apparatus and t the PM . Thus, t2+ on membrane trafficking in animals is the activation of exocytosis during neurotransmission [2+ is also intimately connected to regulated exocytosis, as exemplified in gibberellic acid-induced alpha-amylase secretion [2+ on exocytosis could also be directly observed by Ca2+-induced increases of membrane capacitance in protoplasts of barley aleurone cells [2+ might also increase secretion by stimulating de novo synthesis of secretory cargoes [2+ could be involved in PIN secretion would be a plausible assumption; however, without supporting experimental evidence, it remains equally plausible that PINs are constitutively secreted, in a Ca2+-independent manner. One of the most famous effects of Casmission  and hormsmission . In planecretion ,161, perecretion ,163 and ecretion ,166,167.ne cells ,169, maine cells ,171, maine cells  and tobane cells . Additio cargoes ,174. Tha2+ as a coordinator of PIN trafficking derive from the spatial separation of clathrin-mediated endocytosis and exocytosis of polarized PINs [2+ gradient coordinates secretion, endocytosis and actin dynamics [2+ induces F-actin depolymerization via activation of ABP29 in lily pollen [2+ concentrations, respectively, suggesting that Ca2+ not only stimulates exocytosis, but simultaneously inhibits clathrin-mediated endocytosis. Indications for the involvement of Cazed PINs , which idynamics ,176,177.dynamics . The tipy pollen , therebyy pollen . Therefoy pollen ,184,185.2+ could simultaneously induce secretion and inhibit clathrin-mediated endocytosis of PINs, thereby increasing the local auxin transport capacity, resulting locally in higher auxin concentrations, which, in turn, activate Ca2+ signaling. Such a positive feedback mechanism is consistent with our current models of auxin-regulated PIN polarization [2+ was placed upstream of ROP activity in pavement cells, as Rho GDI1 activity depends on phosphorylation by Ca2+-dependent kinase, CPK3 [2+, as the pollen-specific ROP interactor, RIC3, controls the tip-focused Ca2+ gradient in pollen tubes [Therefore, auxin-induced Carization ,186. Thurization . Upon perization ,78,79,80se, CPK3 . Howeveren tubes .2+ and ROPs are potential mechanisms by which signals, such as auxin [etc., could control the turn-over of PINs to change auxin transport.Such interplays between Caas auxin ,38, cytoas auxin , strigolas auxin , gibbereas auxin ,192, salas auxin , etc., cThe plasma membrane-localized PIN proteins show typical asymmetric distribution patterns ,194 that2+ binding proteins [2+ signals could control PIN polarization via effects on PID activity. Indeed, important Ca2+ signals roughly coincide with PIN polarity changes during phototropism and gravitropism [2+ levels were associated with shifts in PIN polarity [Via misexpression studies, it was found that PIN polarities are not only determined by the cell type, but also by cues embedded in the structure of the PIN itself . These pproteins  suggestsitropism ,205,206.polarity . 2+ is particularly apparent. Gravistimulation induces transient Ca2+ signals in maize coleoptiles [Arabidopsis seedlings [Arabidopsis leaf petioles and hypocotyls [Arabidopsis roots [2+ changes was demonstrated by genetic [2+ sensor, Yellow Cameleon 3.60, a wave of Ca2+ was visualized that spread across the lower side of the root within minutes after gravistimulus [2+ showed dynamics that correlate with auxin transport, but also auxin transport-dependent directional movement of Ca2+ across the gravistimulated tissues could be detected [2+ signals are particularly relevant, as gravitropic bending is severely impaired upon chelation of apoplastic Ca2+, inhibition of calmodulin or Ca2+ channels [2+ in gravitropic bending. However, the underlying molecular mechanism remains poorly understood and, in some cases, even controversial [Gravitropism is an excellent example in which the interplay between auxin and Caeoptiles , whole Aeedlings , Arabidopocotyls  and Arabis roots . The aux genetic  and phar genetic ,210. Viastimulus , correlastimulus . Not onldetected ,213. Thechannels ,218,219.oversial . 2+ signals is the plant’s ability to redirect auxin transport in response to the gravistimulus. Within minutes after graviperception, PIN3 and PIN7 root columella cells repolarize towards the direction of the gravitational pull [+/IAA- symport [Arabidopsis roots reach about the mid-point of bending, the asymmetry in auxin distribution is rapidly lost [A potential target of gravistimulus-induced Canal pull ,221, the symport ,135, to  symport ,222,223. symport ,38. Simu symport ,37. Thesdly lost  and is a2+ and can be summarized in the following hypothetical model: Calcium signaling during graviperception activates PIN3 and PIN7 relocation in the columella to redirect auxin flow to the lower side of the gravistimulated root. Here, auxin induces Ca2+ signals that enhance auxin transport by impacting on trafficking, polarity and, even, the activity of the auxin efflux machinery (+ ATPase activity, resulting in differential growth that is associated with root bending.Nearly every step of this summary of auxin-dependent root gravitropism has been highlighted in the paragraphs before as potentially regulated by Caachinery . MoreoveTIR1/AFB-based auxin signal transduction. Numerous aspects of plant development can now be explained via this pathway and are fully justifying the efforts invested. Yet, it is clear that our understanding of auxin signaling will never be complete by only studying auxin-regulated transcription. Recent work on leaf pavement cell morphogenesis and feedback regulation of auxin transport highlighted the non-transcriptional effects of auxin as important aspects of general auxin physiology. Therefore, it will be of interest to revisit some of these physiological experiments in the context of more recent models of auxin action, armed with a new array of cell biological, genetic and molecular tools to gain more holistic insight into the mechanism of auxin-regulated plant growth and development. Following an era of physiological approaches, the auxin field shifted its attention towards elucidating the mechanisms of auxin-regulated transcription. This move has proven to be very successful with the genetic and molecular characterization of SCF


Bladder Cancer (BC) represents a current clinical and social challenge. The recent studies aimed to describe the genomic landscape of BC have underscored the relevance of epigenetic alterations in the pathogenesis of these tumors. Among the epigenetic alterations, histone modifications occupied a central role not only in cancer, but also in normal organism homeostasis and development. EZH2 (Enhancer of Zeste Homolog 2) belongs to the Polycomb repressive complex 2 as its catalytic subunit, which through the trimethylation of H3 (Histone 3) on K27 (Lysine 27), produces gene silencing. EZH2 is frequently overexpressed in multiple tumor types, including BC, and plays multiple roles besides the well-recognized histone mark generation. In this review, we summarize the present knowledge on the oncogenic roles of EZH2 and its potential use as a therapeutic target, with special emphasis on BC pathogenesis and management. Bladder cancer (BC) displays one of the highest incidences among different tumors in Western Europe. It also has a high prevalence due to the relatively low mortality. BC incidence is 3 times higher among men than women, being the 4th and the 11th most common cancer, respectively . The infin situ), Ta (Non-invasive papillary carcinoma), T1 , T2 (Tumor invades muscularis propria), T3  or T4 (tumor has spread beyond the fatty tissue and nearby organs or structures)), which reveals the location and invasiveness of the tumor. The grade (G1–3) evaluates the differentiation state of the tumor (from highly to poorly differentiated). This classification dates from 1973 and it is the most referenced worldwide [BC is classified depending on the stage (CIS (carcinoma orldwide . This neorldwide .etc.), it is followed by local instillation with chemotherapeutics  [The Non-Muscle Invasive Bladder Cancer (NMIBC) (Ta-T1 stage) is the most frequent form of BC, representing up to 70% of newly diagnosed cases . The othitabine) ,12 or Baitabine) ,14. Thesitabine) ,16. Due itabine) ,18. RemaPI3KCA  and FGFR3 (Fibroblast growth factor receptor 3) [TP53 (Tumor Protein P53) and RB1 (Retinoblastoma 1) genes [Molecular portrait of NMIBC and MIBC has led to the consideration that these two types of tumors arise as a consequence of distinct molecular alterations. In general, NMIBC has been associated with alterations in oncogenes such as eptor 3) ,20,21, w1) genes . These o1) genes ,24. Alth1) genes . The pro1) genes ,26,27 di1) genes .In addition, the genomic characterization of BC samples has allowed the identification of novel pathways. Besides the “usual suspects”, BC is also characterized by the frequent alterations in DNA repair and in chromatin remodeling genes . These oRegarding the chromatin-remodeler genes, the involvement of Polycomb Repression Complex (PRC) has gained relevance in the last years, due to its implication in multiple malignancies ,30,31,32The Polycomb Repressor Complex 1 (PRC1) is primarily responsible of the monoubiquitynation of H2AK119 and of chromatin compaction, and recognizes the H3K27me3 mark left by the PRC2 complex. The core is always formed by the RING1A/B protein. It can bind BMI1 (B lymphoma Mo-MLV insertion region 1 homolog), MEL18 (Melanoma Nuclear Protein 18) or NSPC1 (Nervous System Polycomb-1), and associates with CBX (Chromobox homolog) and HPH (Human PolyHomeotic) proteins. It has also been reported that the complex (RING finger protein 1A/B-BMI1) RING1A/B-BMI1 can form other PRC1-like complexes, whose function remains unclear . As in tThe implication of EZH2 in tumorigenesis has been extensively documented in various types of tumors, including breast and prostate. Of note, its roles include not only epigenetic silencing through histone methylation, but also through gene expression activator of different pathways, and as a modulator of other cell proteins .The EZH2 catalytic activity requires its SET domain . NonetheEZH2 may also cooperate with other epigenetic modifiers, such as DNA methyltransferases (DNMTs)  to promoInterestingly, a novel EZH2 isoform, generated by alternative splicing and named EZH2β, has recently been characterized . This isThe capacity of EZH2 to silence tumor suppressor genes or microRNAs justifies its consideration as an oncogenic factor in multiple cancers ,50. In tet al. (2012) demonstrated that EZH2 phosphorylation in Serine 21 by AKT can switch its functions from transcriptional repression to gene activation [Interestingly, various studies have demonstrated that EZH2 not only acts as epigenetic silencer, but also it can act as a gene activator, independently on PRC2, through methylation of non-histone proteins favoring its oncogenic activity. For instance, in castration-resistant prostate cancer (CRPC), Xu tivation . This motivation . Similartivation . Remarkativation .Other oncogenic activities associated with EZH2-mediated methylation of other non-histone proteins independently of PRC2 include the methylation-dependent nuclear receptor RORα degradation . RORα haEZH2 can also modulate gene expression independently of its histone methyltransferase activity. For instance in breast cancer cells, EZH2 can activate the transcription of different genes in two different manners, depending on the presence or absence of estrogen receptors (ERα). In ERα-positive luminal-like breast cancer cells, EZH2 can form transcription complexes with ERα or its co-activators, facilitating the TCF/β-CATENIN-mediated gene transcription . On the The importance of non-canonical function of EZH2 in BC still needs to be clarified. For instance, there are studies showing the STAT3 implication in MIBC ,69,70, bAll these processes reinforce the role of EZH2 as an oncogenic factor, in particular associated with aggressiveness, progression and worse clinical outcome ,72, as wEZH2 expression can be regulated at multiple levels: it can be transcriptionally induced by multiple factors, such as by E2F family and C-MYC activation ,75,76, oRegarding the regulation of EZH2 expression and/or activity by non-coding RNAs, special interest has been focused on miRNAs and lncRNAs. The miRNAs are small non-coding transcripts that participates in fundamental biological processes including development, apoptosis, differentiation, and proliferation ,88. AccoThe lncRNAs play a central role both in biological processes and pathological events ,92,93. TSeveral miRNAs have been demonstrated to be involved in EZH2 regulation. In et al. (2014) [EZH2 is not only regulated by, but it also regulates a wide variety of miRNAs through epigenetic repression. These miRNAs may act as tumor suppressors, modulating tumor growth, a cancer stem cell phenotype, and cancer cell invasiveness. This EZH2-mediated repression has been demonstrated for miR-31, miR-200a-b-c, miR-181, miR-203, and Let 7b-c ,113,114.. (2014)  reported. (2014) ,117. Int. (2014) ,119. In . (2014) . A simil. (2014)  and in Bet al. (2008) [et al. (2014) [et al. (2009) [Of note, among the miRNAs that regulate EZH2, miR-26a, miR-101 and miR-214 have been proven to be repressed by EZH2. Varambally . (2008)  reported. (2014)  describe. (2014) . In this. (2014) . Similar. (2014) . In the . (2009)  reportedSome miRNAs frequently associated to EZH2 regulation in several tumor types, such as bladder and gastric cancer, hepatocellular carcinoma and neuronal differentiation, include miR-214, miR-124 and miR-137 ,122,123.Long non-coding RNAs (lncRNAs) functions include to induce local gene silencing through binding to chromatin modification complexes, to recruit chromatin modifiers, and to act as scaffolds for chromatin modifying factors changing histone marks thereby modifying gene expression ,127. TheProstate Cancer Associated ncRNA Transcript-1 (PCAT-1) is markedly increased in high-grade and metastatic prostate cancer. Its demonstrated interplay with PRC2 suggests that it could have an important role in prostate cancer progression [Several lncRNAs have been shown to interact with PRC2, facilitating its recruitment to the promoter of some target genes. For instance, the gression . H19. Luo et al. (2013) [H19, already described as an enhancer of tumorigenic potential of carcinoma cells in vivo, promoted BC metastasis inhibiting E-CADHERIN expression and by associating with EZH2 .Another example is the lncRNA . (2013)  demonstrUp-regulated in Bladder Cancer 1 (linc-UBC1) also can be physically associated with PRC2 complex and appeared frequently upregulated in BC. It promotes increased cell proliferation, migration, invasion, metastatic potential, and its increased expression correlates with poor clinical outcome [The lncRNA  outcome .Antisense Non-coding RNA in the INK4 (INhibitors of CDK4) Locus (ANRIL) has been located within the p15/CDKN2B (Cyclin-dependent kinase inhibitor 2B)-p16/CDKN2A (Cyclin-dependent kinase inhibitor 2A)-p14/ARF (ADP Ribosylation Factor) gene cluster. It interacts with EZH2, SUZ12, and the PRC1 subunit CBX7 (ADP Ribosylation Factors) mediating the silencing of p16INK4a, p15INK4b and p14ARF tumor suppressors through the recruitment of PRC2 and PRC1 [EZH2 occupancy levels near the INK4a and ARF genes promoters [The lncRNA and PRC1 ,133,134.romoters . Howeverromoters ,135.EZH2-lncRNA functional interaction is the Homeobox Antisense Intergenic RNA (HOTAIR). It is transcribed in antisense direction from the HOXC (Homeobox C) gene cluster and binds to EZH2 recruiting PRC2 complex to specific target genes genome-wide. Kaneko et al. (2010) [et al. (2010) [HOTAIR to regulate gene expression. HOTAIR is frequently up-regulated in different cancer tissues, in particular in aggressive forms, and associated with metastasis development and poor clinical outcome [HOTAIR expression has also a prognostic value for BC recurrence, progression, and patient survival [HOTAIR in modulating the cancer epigenome have not been completely elucidated. For example, in HOTAIR-deficient mice, the absence of this lncRNA causes H3K4me3 gain, whereas the effect on H3K27me3 is less evident [HOTAIR also interacts with the LSD1 (Lysine-Specific Demethylase 1) H3K4-specific demethylase, these findings may hamper the potential relevance of the HOTAIR-EZH2 interaction in vivo. Whether these effects observed during normal mouse development could also similarly affect carcinogenesis, or if there are species-specific differences attributable to the disparity between mouse and human HOTAIR [The most documented example of . (2010)  and Tsai. (2010)  demonstr outcome ,138,139.survival ,140. How evident . Since Hn HOTAIR , remainsde novo repression but might significantly cause a reduction in the epigenetic plasticity, leading to a permanent repression of key regulatory genes and contributing to tumor development and/or progression. Although PRC2 functions predominantly repressing target genes via the H3K27me3 repressive mark, other epigenetic mechanisms seem to be also involved in this gene silencing mediated by EZH2. Indeed, PRC2 components interact with DNMTs (DNA methyltransferases) and HDACs . The DNMIn this regard, the concerted action of DNMT1 and EZH2 mediates the repression of the miR-200a,b/429 locus contributing to the progression of gastric cancer and glioblastoma . InteresBesides the H3K27me3, other well-characterized histone methylation event is the H3K9me2/3, a hallmark of constitutive heterochromatin. This methylation is catalyzed by specific histone methyltransferases (HMTs), including G9a (EHMT2), GLP (EHMT1) and SUV39H (Suppressor of Variegation 3–9 Homolog). Interestingly, although H3K9me2/3 and H3K27me2/3 marks were first thought to have distinct functions and to be mutually exclusive in genome, recent studies show the co-occupancy of these repressive marks at several gene loci ,146, andOn histone tails, lysine residues can also undergo acetylation, resulting in a more open chromatin configuration . HistoneUTX/KDM6A (Lysine (K)-Specific Demethylase 6A) and JMJD3/KDM6B (Lysine (K)-Specific Demethylase 6B), can particularly remove di- and trimethyl marks from H3K27 and acting against the histone modification mediated by EZH2 [JMJD3/KDM6B in BC. In this regard, opposite functions of KDM6B and KDM6A have been demonstrated in the T-cell acute lymphoblastic leukaemia (T-ALL) [On the other hand, lysine-specific demethylases (KDMs) counteract different histone methyltransferases including EZH2 ,157. Sev by EZH2 ,162,163. by EZH2 , and mos by EZH2 . Importa by EZH2 ,167 and  (T-ALL) , being KAs described above, EZH2 represents an oncogenic signal in several types of cancer, as prostate, liver, breast, colon, skin, lymphoma, endometrial, lung, myeloma, gastric, and BC ,169,170.S-adenosyl-l-methionine (SAM), to the lysine side chains of the acceptor protein. This generates S-adenosyl-l-homocysteine (SAH), which is further processed by the S-adenosyl-l-homocysteine hydrolase (SAH Hydrolase) [The EZH2 protein contains different domains: H1 and H2 domains conform the binding region for PHF1 and SUZ12 respectively; cysteine rich domain, SANT domain to interact with histones; and C-terminal SET domain . The catdrolase) .One of the best characterized EZH2 inhibitors is 3-dezaneplanocin-A (DZNep), an equivalent of 3-deazaadenosine. It blocks the activity of SAH Hydrolase ,189, proin vivo data, D9 is becoming an interesting drug candidate thanks to its anti-cancer effects.This lack of specificity has led to the development of DZNep analogs with improved characteristics. One of them, named compound D9 has shown effectiveness in acute myeloid leukemia cells through the inhibition of various oncogenic signaling pathways . Moreoveet al. (2015) have published other molecule, EPZ-011989, whose difference with EPZ-6438 is a replacement of the second benzene ring and pyridine residue to increase the range of EZH2 inhibition, improving the pharmacokinetics and pharmacodynamics qualities, at least in a mouse xenograft model of human B cell lymphoma [Various inhibitors have been developed to achieve the competitive inhibition of the SAM binding to the SET domain of EZH2 . These ilymphoma .Other SAM-competitive inhibitors are EI1 from Novartis , and GSKA dual inhibitor of EZH2 and EZH1 (UNC1999) has also been developed . It is sConstellation Pharmaceuticals have developed various compounds with potential EZH2 inhibitory activity. After a general screening in search for potential inhibitors inducing genome-wide histone H3K27me3 reduction with activity at nanomolar concentration, they identified the lead compound CPI360 . CPI-360in vivo. Indeed, while they are easily internalized by the cells in vitro, an in vivo investigation is still needed. In addition, as mentioned above, some EZH2 functions are independent on the binding to other PRC2 components. Consistently, these peptidomimetics approach will not impair these activities.The ability of SAH to inhibit the catalytic activity of EZH2 has been exploited by various groups trying to avoid the general effects attributable to overall SAH accumulation. One of them consists on the generation of stabilized peptides mimicking the α-helix of EZH2. This domain recognizes the corresponding domain of EED, therefore promoting the dissociation of the EZH2/EED complex, and impairing the function of PRC2 . DetaileA series of compounds, also based on the structure of SAH, have been recently developed by Pfizer researchers . Importain vivo [As mentioned above, EZH2 is also regulated by proteasome-mediated proteolysis ,79,199. in vivo . NonetheFinally, another potential approach is to induce the expression of miRNAs targeting EZH2. Such approach has been reported for the natural product derivatives Gambogic Acid (GA) and Methyl Jasmonate (MJ). They are both considered promising therapeutic agents against various cancer types including prostate, pancreas, gastric, breast, and lung cancer, and their combination causes synergistic inhibitory activity. Interestingly, in a recent detailed study focused on BC cells, it has been shown that MJ significantly sensitized BC cells to GA-induced growth inhibition and apoptosis . The plaThe role of EZH2 in chemotherapy resistance is to be determined in BC, although recent studies showed that acquired cisplatin resistance in ovarian cancer appeared linked to overexpression of EZH2 ,203, in in vivo using genetic technologies [EZH2 plays a main role on the maintenance of cellular epigenetic integrity, being highly relevant on human BC, among other tumors such as prostate, breast, ovarian, renal carcinoma, lung, liver, brain, gastric, esophageal, pancreatic, or melanoma. As a main player within cancer epigenetics, it can become a promising tool for detection, diagnosis, prognosis, and prediction of response to possible therapies. Nevertheless, and in spite of the wide knowledge on EZH2 in normal development and in pathological conditions, there are still a large number of unanswered questions, which might be particularly relevant in BC and other cancers, where EZH2 upregulation appears to dictate the malignant progression. For instance, it is mandatory the identification of EZH2 target genes that are directly responsible for such oncogenic progression. In addition, it is interesting to determine whether EZH2 functions independent on histone methyltransferase are of relevance in these tumors. Similarly, whether the inactivation of KDM6A is equivalent to EZH2 overexpression is still unknown. There is still paucity in studies of EZH2 inhibitors in solid tumors, including BC. In these studies, the reported differences between long term EZH2 depletion as antitumoral approach nologies  or shortnologies . AnotherIn summary, although our knowledge on the molecular mechanisms of EZH2 in relation with malignant transformation has been greatly improved in the recent years, there is still a lot of missing information necessary for full establishment of clinical settings of using this molecule as a target for cancer patients, in particular in BC.


Individuals with systolic heart failure are at risk of ventricular arrhythmias and all-cause mortality. Little is known regarding the mechanisms underlying these events. We sought to better understand if oxylipins, a diverse class of lipid metabolites derived from the oxidation of polyunsaturated fatty acids, were associated with these outcomes in recipients of primary prevention implantable cardioverter defibrillators (ICDs).Among 479 individuals from the PROSE-ICD study, baseline serum were analyzed and quantitatively profiled for 35 known biologically relevant oxylipin metabolites. Associations with ICD shocks for ventricular arrhythmias and all-cause mortality were evaluated using Cox proportional hazards models.1α  were associated with the risk of appropriate ICD shock after multivariate adjustment for clinical factors. Additionally, 4 oxylipin-to-precursor ratios, 15S-HEPE / FA (20:5-ω3), 17,18-DiHETE / FA (20:5-ω3), 19,20-DiHDPA / FA (20:5-ω3), and 5S-HEPE / FA (20:5-ω3) were positively associated with the risk of all-cause mortality.Six oxylipins, 17,18-DiHETE , 19,20-DiHDPA , 5,6-DiHETrE , 8,9-DiHETrE , 9,10-DiHOME , and PGFIn a prospective cohort of patients with primary prevention ICDs, we identified several novel oxylipin markers that were associated with appropriate shock and mortality using metabolic profiling techniques. These findings may provide new insight into the potential biologic pathways leading to adverse events in this patient population. In the US, approximately 3 million people have heart failure with reduced ejection fraction and are potentially eligible for implantation of a defibrillator (ICD) for primary prevention of sudden cardiac death (SCD) –2. HowevThe metabolic milieu of cardiomyocytes, both from normal and diseased hearts, contributes to the generation of arrhythmias associated with SCD –11. ReceOxylipins, a diverse class of lipid metabolites derived from the oxidation of polyunsaturated fatty acids, are potent endogenous signaling molecules involved in the regulation of various metabolic processes such as inflammation, thrombosis, lipid management, blood pressure regulation, and hemostasis –19. In aThe Prospective Observational Study of Implantable Cardioverter-Defibrillators (PROSE-ICD) is a multicenter study of systolic heart failure patients receiving a primary prevention ICD. Enrollment was conducted at four United States clinical centers from 2003 to 2013 . Among tAt enrollment, all participants underwent a comprehensive medical history, cardiovascular examination, and fasting blood collection . PeripheAfter enrollment, patients were evaluated (either in person or by phone) every 6 months and after any patient-perceived ICD shock. For this analysis, patients were followed for events through June 1, 2015. The primary endpoint was appropriate ICD shock defined as a shock delivered for rapid ventricular tachyarrhythmias. The secondary endpoint was all-cause mortality, which was obtained by next-of-kin phone interviews and National Death Index queries. Arrhythmic events were adjudicated by two cardiac electrophysiologists blinded to patient information. Disagreements were reconciled by a third electrophysiologist.Methodologies for oxylipin analysis have been previously described . For oxyMetabolites with levels below the lower limits of detection were imputed with a value of the lower limit of detection (for the given compound) divided by 2. Metabolites with a significant amount of missing values (>25%) were excluded from the analysis. Due to the skewed distribution of the oxylipins, values were log-transformed to approximate a normal distribution and subsequently standardized to have a mean of 0 and standard deviation of 1. For each oxylipin, we also calculated the oxylipin-to-precursor ratio as the ratio may provide information above and beyond their individual components . Similar to oxylipins, all oxylipin-to-precursor ratios were also log-transformed and standardized to have a mean of 0 and standard deviation of 1.To identify associations of each oxylipin and oxylipin-precursor ratio with the study endpoints, we used the Cox proportional hazards regression to calculate the hazard ratios (HR) associated with every standard deviation (SD) change in the log-transformed oxylipin level. For all analyses, we used two models with covariates added sequentially. The first model was adjusted for age, sex, race, and enrollment center. The second model was further adjusted for ejection fraction, NYHA class, cardiomyopathy etiology, atrial fibrillation, diabetes, hypertension, and CKD. We also performed sensitivity analyses further adjusting for ECG markers  and medications , and found no significant differences in the results (data not shown). Since the nature of this analysis is largely exploratory, nominal p-values from the Cox regression models were reported. STATA version 12  was used for all analyses.The average age of participants at baseline was 60.1 ± 12.8 years. Men comprised of 72.9% of the study population and 62.6% were Caucasians . After a1α  were associated with the risk of appropriate ICD shock after adjusting for age, sex, race, enrollment center, smoking status, body mass index, ejection fraction, NYHA class, atrial fibrillation, diabetes, hypertension, and chronic kidney disease , 19,20-DiHDPA , 5,6-DiHETrE , 8,9-DiHETrE , 9,10-DiHOME , and PGF disease . None ofWe found similar patterns of associations between oxylipin-precursor ratios and the risk of appropriate shocks for the oxylipins mentioned above . Additio1α) that were associated with the risk for appropriate ICD shocks in a prospective cohort of primary prevention ICD patients, suggesting a role as novel markers of ventricular arrhythmias. Additionally, the ratios of 4 oxylipins to their precursors were positively associated with all-cause mortality. These associations remained true even after adjustment for several demographic variables.Using metabolic profiling, we identified 6 oxylipins , linoleic acid (LA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and dihomo-γ-linolenic acid (DGLA) . DiffereAlthough the role of many oxylipins in cardiovascular disease is still uncertain, accumulating evidence suggests that they play an important role in the progression of cardiovascular risk factors, inflammation, and thrombosis , 19, 22.2+ overload [2+ sensitivity in human pulmonary arteries [Oxylipins 9,10-DiHOME, 17,18-DiHETE and 19,20-DiHDPA are stable metabolites of LA, EPA and DHA, respectively, that are produced through the CYP450 pathways . The actoverload . Additioarteries . Our stu1α is a biosynthesis product of DGLA via the COX pathway [1α could modulate the contraction and relaxation of vascular smooth muscle from the arterial strips [1α intravertebrally caused an increase of blood pressure and tachycardia, whereas intravenous or intracarotid infusion of the same dose of PGF1α had no effect. These findings suggest that PGF1α has little direct effect on heart rate but may affect heart rate indirectly through a reflex stimulation or inhibition on the sympathetic and vagal nerves [1α and appropriate shock is unknown.PGF pathway . In our l strips  and incrl strips . In an al nerves –41. BeyoAdditionally, we found positive associations between the ratios of 5S-HEPE and 15S-HEPE to their respective precursors and the risk of mortality. Both 5S-HEPE and 15S-HEPE were metabolites of EPA via the LOX pathway . These LSeveral limitations of our study need to be considered. Since our study was by nature observational, we could only identify associations but not establish causal links between oxylipins and outcomes. Although our study included a large cohort of systolic hear failure patients with primary prevention ICDs, it may be still underpowered to detect associations with outcomes as relatively few patients experienced these events. Given the nature of our cohort, findings from this analysis may not be applicable to patients at high risk for sudden death but with preserved left ventricular function.In a prospective cohort of patients with primary prevention ICDs, we identified several novel oxylipin markers that were associated with appropriate shock and all-cause mortality using metabolic profiling technique. Additional studies are required to confirm these findings in other patient populations and to better understand the exact mechanisms underlying the associations between oxylipins and SCD.


Ochrobactrum sp. SJY1 uses nicotine as the sources of carbon, nitrogen, and energy. The genome of SJY1 was sequenced in order to provide insights into its mechanism of nicotine degradation. Physiological characteristics and genome analysis indicate that strain SJY1 might have a different nicotine degradation pathway from the pyridine or pyrrolidine pathway. Pseudomonas putida S16 with the pyrrolidine degradation pathway, Arthrobacter sp. M2012083 with the pyridine pathway, and Pseudomonas geniculata N1. Recently, a Gram-negative strain, Ochrobactrum sp. SJY1, which uses nicotine as the sole source of carbon, nitrogen, and energy, was newly isolated from water. Strain SJY1 was deposited at the China Center for Type Culture Collection under accession number CCTCC AB 2014146. The previously isolated Ochrobactrum bacterium, O. intermedium DN2, was reported to have the capability to degrade nicotine  using Velvet 1.2.10 software (The genome of strain SJY1 was sequenced by an Illumina HiSeq-2000 sequencer (2- × 101-bp paired end). The total amount of read data comprises 1,374,835,836 bases. The reads were assembled moaACDE, mobAB, modABC, moeAB, and mog) for the synthesis of molybdenum cofactors and the genes from downstream of the pyrrolidine pathway  were found in the genome of strain SJY1 (Pseudomonas sp. HZN6 (Thirty-six CDSs for denitrification were annotated, indicating strain SJY1 may come from anaerobic conditions. Several genes (ain SJY1 . Howeversp. HZN6 , 11, 12.AZRT00000000. The version described in this paper is version AZRT01000000.This whole-genome shotgun project has been deposited at DDBJ/EMBL/GenBank under the accession number


Inositol hexakisphosphate is known to be the phosphorous reserve in plants particularly in the seeds. Though ithas been known for its antinutrient properties for many years, recent research shed light to reveal it as a novel anticanceragent. Hence the present study investigates the drug-likeness of phytic acid and its analogues through bioinformaticsmethods. Two potential cancer drug targets such as mitogen activated kinase and inositol 1,4,5-triphosphate receptor areincluded in the study. Out of 50 selected analogues of phytic acid, 42 structures interact well with the chosen drug targets.The best interacting structures are 1-diphosinositol pentakisphosphate and 2,3,4,5,6-pentaphosphonooxycyclohexyldihydrogen phosphate. For both of these structures, the negative binding energy obtained was -49.5 KJ/mol; this affirmsthe stability of the complex. ADME properties are also predicted to assess the drug-like properties of the compounds. Thestructure activity relationship model is generated for 12 compounds with experimental IC50 values.  At present there is no published research work on the structure-activity relationship (SAR) of phytic acid and its analogues. Elucidating SAR can help in screening out novel inhibitors that can bind with the druggable targets. Hence the present study makes an attempt to bring forth the drug-like properties of phytic acid and its analogues through computational approach.Inositol hexakisphosphate InsP6), also known as phytic acid is the foremost inositol phosphate discovered . It is tal salts  and thei, also knal salts , 8. The al salts . Besidesfighter’ . However22.1The 2D structures of phytic acid and its analogues were retrieved from the Pubchem repository of NCBI . LigprepCID477, CID890, CID107758, CID125004, CID127297, CID178749, CID439456, CID443266, CID2524165, CID4200706, CID4487899, CID10251645, CID10747577, CID14375662, CID16752671, CID16752673, CID18365880, CID21099914, CID44274820, CID44332437, CID45479488, CID46173206, CID46173281, CID46173316, CID46173429, CID46173525, CID46905360, CID52949527, CID53379838, CID53380009, CID53380097, CID53380098, CID53380099, CID53380100, CID53380197, CID53380199, CID53380200, CID53380300, CID53380301, CID53380721, CID53380834, CID53380835, CID53380836, CID53380837, CID53462026, CID53477671, CID54002314, CID54207372, CID57503246 and CID57773931.2.2Identification of a protein target is a crucial step in interaction studies. In the present work one such target was selected using PASS online server . The ser2.3Grid generation was done using the Glide module of Schrödinger Release 2013-1 . The lig2.4The ligand binding energies were calculated using molecular mechanics with generalized Born and surface area solvation (Prime MM-GBSA) . Combinaenergy, mmgbsa energy, mmgbsa energy of the free ligand, mmgbsa energy of the uncomplexed receptor, mmgbsa energy of the ligand in the complex, mmgbsa ligand strain energy and mmgbsa free energy of binding excluding ligand strain.2.5Absorption, distribution, metabolism, and excretion (ADME) properties of phytic acid and its derivatives were predicted by QuikProp . QuikPro2.650 values were available in Pubchem database. Based on the training set, IC50 values were predicted using quantitative neighbourhood of atoms (QNA model) and multilevel neighbourhood of atoms (MNA model) [Quantitative structure activity relationship of phytic acid and its derivatives were predicted by using GUSAR software. It helps to create QSAR models on the basis of the appropriate training sets . It usedA model) . On the 31. It is instinctive as well as evident that phytic acid is an inositol 1,4,5-triphosphate receptor 1 antagonist. This is in concord with Sakakura et al., they suggested inositol 1,4,5-triphosphate receptor as a molecular target for gastric cancers [3 is available in PDB (1N4K). Inositol 1,4,5-triphosphate receptor is involved in calcium signalling process. Calcium signaling is a vibrant signalling pathway which regulates numerous cellular processes, including fertilization, cell growth, transformation, secretion, smooth muscle contraction, sensory perception and neuronal signaling [proliferation, gene expression, differentiation, mitosis, cell survival, and apoptosis [The structure of phytic acid was very likely to be active against 46 targets, Table  cancers . The cryignaling . Bindingpoptosis . There apoptosis , 32. 3.12 and Table 3 show the top ten docked poses. Out of fifty compounds, for 1PMQ forty two gave docking scores less than -10 and for 1N4K forty scored less than -10 which shows both the protein targets have good binding affinity towards the selected compounds. All the ligands which scored less than -10 have hydrogen bond donor atom count ranging from 8 to 14, hydrogen bond acceptor atom count ranging from 12 to 30 and heavy atom count ranging from 24 to 44 according to Pubchem repository [Docking was carried out using phytic acid and its analogues using the targets, 1N4K and 1PMQ. The results show that all the selected compounds effectively bind with the selected targets. 1-diphosphoinositol pentakisphosphate (46173525) gave the best docking score of -14.29 for 1PMQ and 2,3,4,5,6-pentaphosphonooxycyclohexyl dihydrogen phosphate (178749) gave the best docking score of -14.02 for 1N4K. Table pository . These fpository . Chiral pository . This ex1).The amino acids involved in the binding site of 1N4K include tyrosine 567, arginine 511, arginine 265, lysine 508, arginine 269, glycine 268, lysine 569, arginine 568 and threonine 267. The amino acids involved in the binding site of 1PMQ include lysine 93, tyrosine 223, lysine 191, valine 225, glutamine 255, alanine 74, aspartate 189, valine 224, arginine 107, threonine 226, serine 193, serine 72, glycine 71, valine 78 and aspargine 194. The amino acids in the binding pocket of both the proteins have some residues in common, like tyrosine, arginine, threonine, lysine and glycine. Glide scoring function is given by the equation XP GlideScore = Ecoul + EvdW + Ebind + Epenalty , 36 wher3.24). The free energy of binding was least for -2,3,4,5,6-pentakis (phosphonooxy) cyclohexyl tetrahydrogen triphosphate (53477671). This ligand has 8 phosphate groups available for binding when compared with others. It also pocess maximum numbers of hydrogen bond donor atom count, acceptor atom count, heavy atom count and complexity values of 14, 30, 44 and 1340 respectively [-1), van-der-waal’s energy , co-valent energy , coulombic binding energy  and solvation binding energy  were also significantly low. The presence of aromatic group and hydroxyl group bearing tyrosine, phosphate anion binder arginine and highly reactive amino group bearing lysine in the binding pocket of both the targets also account for the binding efficacy and binding energy minima [-1. This compound is otherwise known as technetium-99m. Technetium-99m is a radioisotope most widely used in medicine employed in over half of all nuclear medicine procedures [The result of Prime MMGBSA distinguishes strong and weak binders , hydrophobic and hydrophilic component and volume were obtained. The Table le bonds . 3.46. For QNA model and Combinatorial model the predicted IC50 values were very less deviated from the experimental IC50. QNA model gave r2 (goodness of fit) of 0.943 and Combinatorial model gave the best result with r2 value 0.960. For MNA model the r2 value is 0.660.QSARs are mathematical models that attempt to relate the structure-derived features of a compound to its biological or physicochemical activity. QSAR works on the assumption that structurally similar compounds have similar activities . In the 2, q2, F and low value of SD and the equation indicates that the model is statistically significant, could be used for analogue screening. The regression equations for the MNA model, QNA model and combinatorial model are 1.33x-7.298, 0.899x + 0.419 and 0.836x – 0.303 respectively indicates a linear relationship between experimental IC50 value and predicted IC50 value. Therefore phytic acid and the selected analogues for GUSAR studies have similar activities. The statistical characteristics of the model with high value of rThe present work identified mitogen activated kinase and inositol 1,4,5-triphosphate receptor as the protein targets for phytic acid. The interaction (docking scores) are -14.29 and -14.02, respectively. The protein ligand complexes had a minimum binding energy of -49.5kJ/mol. The parent compound, phytic acid and its similar compounds resemble similar mode of binding due to chemical similarity. The amino acid pattern involved in the binding interaction is almost same for all the compounds. The analyzed compounds are predicted to be drug-like. The structure activity relationship model correlated the atom level contributions to the activity. This would be made beneficial for designing novel drug compounds. Furthermore animal trials and clinical studies and can reveal more insights into the drug-likeness of phytic acid.


AbstractCobitistakenoisp. n., is described based on the holotype and ten paratypes collected from Tango District, Honshu Island, Japan. The new species is distinguished by a combination of the following character states: 1) the lamina circularis at the base of the pectoral fin in adult male having a simple roundish plate form; 2) a narrowing of the upper segments of the first branched ray of the pectoral fin; 3) a short maxillary barbel whose length equals diameter of the eye; 4) 14 prepelvic myotomes, and 5) L3 and L5 well developed, forming longitudinal obvious stripes in males during the spawning season.A new species of spined loach,  Cobitis Linnaeus, 1758 (Cypriniformes: Cobitidae) includes small, slender-bodied benthic freshwater fishes. The genus is characterised by the following features: the suborbital spine is erectile; the mouth is small and inferior with three pairs of barbels; body pigmentation is organised in one dorsal and four lateral longitudinal lines or rows of blotches; and the presence of the lamina circularis at the base of the pectoral fin in adult males  Cobitisbiwae Jordan & Snyder, 1901, 2) Cobitisstriata Ikeda, 1936, 3) Cobitismatsubarae Okada & Ikeda, 1939, 4) Cobitistakatsuensis Mizuno, 1970, 5) Cobitisshikokuensis Suzawa, 2006, 6) Cobitismagnostriata Nakajima, 2012, 7) Cobitisminamorii Nakajima, 2012, 8) Cobitiskaibarai Nakajima, 2012 and 9) Cobitissakahoko Nakajima & Suzawa, 2015, and six subspecies, namely 1) Cobitisminamoriitokaiensis Nakajima, 2012, 2) Cobitisminamoriioumieneis Nakajima, 2012, 3) Cobitisminamoriiyodoensis Nakajima, 2012, 4) Cobitisminamoriisaninensis Nakajima, 2012, 5) Cobitisstriatafuchigamii Nakajima, 2012, and 6) Cobitisstriatahakataensis Nakajima, 2012 have been described in Japan  was defined as the number of muscle segments between the base of the pectoral fin and the origin of the pelvic fin  were observed and sketched using a stereomicroscope. The dorsal and lateral colour patterns were organised in five longitudinal lines of pigmentation, which were abbreviated as lines L1 to L5 according to the scheme of I examined 11 specimens collected from a small river in Tango District, Kyoto Prefecture, Honshu Island, Japan Figs . There ilvic fin . The rigPageBreakPageBreak – the Kanagawa Prefectural Museum of Natural History, Odawara, Kanagawa, JapanKPM;  – the Tokushima Prefectural Museum, Tokushima, JapanTKPM;  – the Faculty of Agriculture, Kinki University, Nara, JapanKUN and  – private collection of the author, JapanJNC.The type series were deposited in the following collections: Taxon classificationAnimaliaCypriniformesCobitidaehttp://zoobank.org/13895F3E-53D6-4C24-BE3B-E6DBDF7BEF07CobitistakenoiCobitisstriata: Cobitis sp. 5: Cobitis sp.: Cobitis sp.:  ‘Tango tetraploid form’ of KPM-NI 31994, 60.4 mm standard length (SL), male, Tango District, Kyoto Prefecture, Honshu Island, Japan; collected by K. Tominaga on 17 Apr. 2010. Paratypes: 10 specimens, all from same locality as the holotype: TKPM-P 7363, 7364, 53.2–67.5 mm SL, male and female, same data as holotype; KPM-NI 31995–31999, 49.4–70.5 mm SL, 3 males and 2 females, collected by J. Nakajima on 12 Nov. 2010; KUN-P 45133, 57.7 mm SL, male, collected by K. Tominaga on 5 Jul. 2014; JNC 188, 189, 58.6–60.6 mm SL, 2 males, same data.Holotype: USP narrow; PMN 14; line L5 organised in 11–17 oblong or ovoid blotches out of spawning season, and lines L3 and L5 in adult male well-developed longitudinal obvious stripes during spawning season; upper and lower spot at caudal base not connected; tetraploid.Maxillary barbel short, more of the same eye diameter; lamina circularis in adult males simple and roundish; PMN 14. Very small cycloid scales on the trunk. Suborbital spine two-pronged and incurved; length of the outer spine one-third of that of the inner spine ; caudal vertebrae 20 (19–21); total vertebrae 42 (40–44)  . Largestn female .PageBreakof the head covered with amorphous spots; opercle and snout covered with amorphous patterns. Caudal and dorsal fins with 3–4 arcuate bars. Anal fin pigmented along fin rays. Upper spot at caudal base jet-black, size comparable to the eye diameter, lower spot at the caudal base relatively inconspicuous and small; upper and lower spots at the caudal base not connected. Male out of spawning season  by a combination of the following character states: a short maxillary barbel equaling in length the eye diameter ; a simple roundish lamina circularis ; a narrow USP ; PMN 14 ; a L5 formed of blotches out of spawning PageBreakseason (vs. stripe-like in and out of spawning season in Cobitistakatsuensis and Cobitismagnostriata); both spots at caudal base obvious (vs. lower spot inconspicuous in Cobitisstriata and Cobitiskaibarai); and ploidy tetraploid . These comparative data were summarised from This new species is distinguished from nine species of Cobitistakenoi has only been found in one small river system, and the habitat is under threat from river improvement. In addition, some threatened freshwater fishes are captured and sold illegally in Japan  – as Cobitis sp. – in the Japanese Red List (Till date, Red List . The dis