The raw data were also analyzed by GeneSpring GX software version 7. 3. 1. The correlation coefficients of gene probes expressed between any two samples were sellckchem cal culated from the normalized values by using GeneChip Robust Multiarray Average algorithm. It may be noted that Affymetrix GeneChip expression analysis can be used as a stand alone quantitative comparison, since the correlation between Affymetrix GeneChip results and TagMan RT qPCR results was shown in a good line arity of R2 0. 95 by the MicroArray Quality Control Study, a collaborative effort of 137 scientists led by the US FDA. A hierarchical clustering and principle component analysis of the eight Affymetrix Gene Chip data from duplicates of four populations of hES cells were also performed in order to check the quality of microarray results.

Analyses of signaling pathways and GO process networks The abundantly expressed mRNAs of T3 HDF and T3 CMHD, as well as T3 MEF and T3 CMMEF, cells were analyzed for signal ing pathways and GO process networks by using Meta Core Analytical Suite as previously described. The MetaCore includes a curated database of human protein interaction and meta bolism, and thus it is useful for analyzing a cluster of genes in the context of regulatory network and signaling pathways. Quantification of miRNAs The expression levels of 365 human miRNAs from T3 HDF and T3 CMHD cells were determined using the TaqMan MicroRNA Assays. The detailed procedure for miRNA quanti fication was previously described. In brief, TagMan MicroRNA Assays include two steps, stem loop RT fol lowed by real time PCR.

Each 10 ul RT reaction that includes 90 ng total RNA, 50 nM stem loop RT primers, 1�� RT buffer, 1. 25 mM each of dNTPs, 0. 25 U ul RNase inhibitor, and 10 U ul MultiScribe Reverse Transcriptase was incu bated in the PTC 225 Peltier Thermal Cycler for 30 min each at 16 C and at 42 C, followed by 5 min at 85 C, and then held at 4 C. RT products were diluted twenty times with H2O prior to setting up PCR reaction. Real time PCR for each miRNA was carried out in triplicates, and each 10 ul reaction mixture included 2 ul of diluted RT product, 5 ul of 2�� TagMan Universal PCR Master Mix and 0. 2 uM TagMan probe, respectively. The reac tion was incubated in an Applied Biosystems 7900HT Sequence Detection System at 95 C for 10 min, followed by 40 cycles of 95 C for 15 sec and 60 C for 1 min.

The threshold cycle is defined as the fraction cycle num ber at which the fluorescence exceeds the fixed thresh old of 0. 2. Total RNA input was normalized based on the Ct values of the TagMan U6 snRNA assay as an endogenous control. The fold change was calculated as 2 CT �� K, where CT ? and K is a constant. 2D gel analysis of proteins Approximately Brefeldin_A 1 �� 106 hES cells on 10 cm plate were washed twice each with 1�� PBS and cell wash buffer, and then lyzed using NP40 lysis buffer. 1 mL ice cold acetone 11% w v trichloroacetic acid 20 mM DTT was added per 0.

Although no information is lost using this method, it is visually inelegant and obscures the model. Another method which preserves the clarity of the ori ginal merely model is to simply keep the original edge types. To make the model executable, we can assign distinct values to each edge type from 0 to m, where m is the number of distinct edge types. Then the graph can be represented by the following sort of adjacency matrix. Set the ijth entry of the adjacency matrix to be 2e taken over edges with type e between vertices i and j. Note that a non directional edge e between vertices i and j will contribute to both the ijth entry and the jith entry, whereas a directed edge e from i to j will only contribute to the ijth entry of the adjacency matrix.

A graph can be reconstructed from such an adjacency matrix under the assumption that the graph does not contain multiple edges of the same edge type. For biological models, the restriction to such graphs is natural. Because our main motivation for using hierarchical graphs to model biochemical net works is to improve the clarity of models, we recom mend this second method. For instance, consider the chemical species graph of Lck with SH2 connected to phosphorylated Y505. Let the hierarchy edges be edge type 0 and the bond edges be edge type 1. Then in the adja cency matrix, a hierarchical edge from i to j will be represented by a 1 20 in the ijth entry, whereas a bond edge between vertices i and j will be represented by a 2 21 in both the ijth and the jith entries. The adjacency matrix is given in Table 1.

As can be seen from the first row of the matrix in Table 1, Lck contains SH3, SH2, Y505 and PTK components. Likewise, from the third and fourth rows, one can see that the SH2 component contains a Y192 subcomponent and the PTK component contains a Y394 subcomponent. From the pair of 2 entries, one can see that there is a bond between the SH2 and Y505 components. An important capability of BioNetGen is the ability to distinguish between different graphs and to recognize isomorphic graphs. We describe a slight generaliza tion of the Nauty algorithm which can canonically label graphs with several edge types. This algorithm, HNauty, has been incorporated into BioNetGen. Although our algorithm is only slightly different from the one described by McKay, we provide a brief description of the whole algorithm for clarity.

Although the representation of graphs within comput ing systems can vary, it is useful to think of a graph as being represented by an adjacency matrix for the graph. However, the same graph can have several different adjacency matrices associated with it, different permuta tions of the vertices may correspond to GSK-3 different adjacency matrices. If a graph is represented by an adjacency matrix, the problem of finding a canonical label for a graph is thus nothing more than picking a canonical adjacency matrix for each graph, that is a canonical permutation of the vertices.

Genes encoding JAK STAT pathway members, includ ing Jak1 and Stat1, were found to be upregulated in our study, suggesting that the JAK STAT pathway may be affected by bacterial infection, which may result in changes in other cross talk biological processes, such as NF B signaling selleck compound pathway, TGF b activated SMAD pathway, and apoptosis. Another signaling pathway affected by bacterial infec tion in the large yellow croaker was the MAPK cascade. This pathway has been demonstrated to regulate the expression of genes involved in the immune response to pathogens, cell differentiation, and cell death. Modulation of MAPK activity in the common periwin kle in response to Escherichia coli derived LPS has been studied. Some key MAPK related genes were identi fied in our transcriptome, including Casp9, Rac1, Gadd45a, and Dusp7.

Quantitative PCR analysis confirmed the differential expression of Casp9 and Dusp7. The Rho family GTPase Rac1 has been implicated in the control of the p38 MAPK signaling pathway by controlling b1 integrin. As shown in humans, dominant negative Rac1 completely inhibits b1 integrin induced p38 MAPK acti vation, whereas wild type Rac1 overexpression causes a slight increase in b1 integrin induced p38 MAPK activa tion. Dual specificity phosphatases including Dusp7 are a subset of protein tyrosine phosphatases, many of which dephosphorylate threonine and tyrosine residues on MAPKs and hence are also referred to as MAPK phosphatases. The regulated expression and activity of DUSP family members in different cells and tissues control MAPK intensity and duration to deter mine the type of physiological response.

There fore, the identified changes in gene expression in the large yellow croaker may facilitate the activation of the MAPK pathway and protect hosts against A. hydrophila infection. Adaptive immunity is the process that leads to specific host resistance to infection. T cells orchestrate responses against such foreign pathogens as viruses and bacteria. TCR and its downstream signaling cascades play a key role in these events. Here, we identified TCR pathway related genes that were downregulated at 24 h after A. hydrophila infection. This complex process is shown in Figure 4, and genes expressed differentially are listed in Additional file 7, Table S7.

Lyn, Itk, Was, Ptpn6, and Jun expression was downregulated, implying that the TCR signaling pathway may be suppressed in the early period following bacterial infection. Stu dies have shown that a fine balance exists between a positive signal that initiates TCR cascade Dacomitinib and a negative signal that controls the threshold, extent, and termina tion of TCR activation. Several protein tyrosine phosphatases have been shown to function as negative regulators of the TCR signaling pathway by dephosphorylating activated signaling molecules. Here, expression of Ptpn6, a member of the PTP family, was downregulated, suggesting that although the TCR signaling pathway was suppressed by A.

A group of research use only German researchers reported that SOCS1 has a nuclear localization signal and is predom inantly localized in the nucleus, unlike CIS 1 and SOCS3. In the nucleus, NF ��B p65 bound to SOCS1 is degraded via ubiquitination with suppression of NF ��B dependent gene e pression. Indeed, in the present study, SCOS1 was present in the nucleus as well as in the cyto plasm of chondrocytes. In addition, NF ��B luciferase activity levels were reduced in the SOCS1 overe pressing cells in the presence of IL 1B. In this conte t, the inhibi tory effects of SOCS1 on the IL 1B induced MMP pro duction may be partially mediated by degradation of p65. However, p65 or phosphor p65 levels did not change with SOCS1 overe pression. Instead, the deg radation of inhibitory I��B was suppressed in the SOCS1 overe pressing chondrocytes after stimulation with IL 1B.

These findings are in line with previous findings that LPS induced I��B degradation was de layed in the SOCS1 transfected RAW264 cells. However, as shown in Figure 7, the antagonistic effect of SOCS1 on IL 1B signaling might not necessarily depend on the downregulation of the NF ��B pathway in human chondrocytes. SOCS1 operated in both MAPK and NF ��B pathways in our study. TAK1 is a kinase that activates both I��B kinase and MAPK kinases, and its activation leads to phosphorylation of p38, JNK, and ERK kinases and I��B degradation. Frob se et al. found that SOSC3 inhibited IL 1B signal transduction via suppres sion of the TRAF6 ubiquitination that is required for TAK1 activation.

However, we did not observe any change in phosphorylation levels of TAK1 in the SOCS1 overe pressing cells. Rather, SOCS1 decreased the levels of TAK1 protein. The dose dependent suppression of TAK1 protein was additionally confirmed by using a transient SOCS1 overe pression system. The SOCS bo is a C terminal domain of SOCS family proteins, including SOCS1, and it is essential to recruit the ubiquitin transferase system. The domain can function as E3 ubiquitin ligases and mediate the ubiquitination and subsequent degradation of target proteins. Thus, we e amined the amount of ubiquitinated TAK1 in the SOCS1 overe pressing chondrocytes and found that ubiquitinated forms of TAK1 were easily detectable after IL 1B stimulation. Moreover, MG132 proteasome inhibitor increased TAK1 levels in SOCS1 overe pressing chondrocytes.

These findings suggested that SOCS1 provides a novel negative feedback mechanism through the degradation of TAK1, which is involved in IL 1B signaling. Although the present study is the first to describe a novel role of SOCS1 in OA pathogenesis, this study has several limitations. First, we used an SOCS1 overe pres sion and knockdown system. GSK-3 Although the SOCS1 e pression is increased in OA chondrocytes in vivo, the SOCS1 in vitro transfection could be overe pressed in supraphysiologic concentrations.

This procedure was used to investigate the effects of GF10903 added selleck chemicals U0126 to the cell suspensions 8 min before activation, on the rate and magnitude of Ca2 influ . Radiometric assessment of Ca2 flu es 45Ca2 was used as tracer to label the intracellular Ca2 pool and to monitor Ca2 flu es in resting and PAF stimulated neutrophils. In the assays of Ca2 influ and efflu described below, the radiolabeled cation was used at a fi ed, final concentra tion of 2 Ci. ml 1, and the final assay volumes were 5 ml containing a total of 1 107 neutrophils. The standardiza tion of the procedures used to load the cells with 45Ca2, as well as a comparison with oil based methods for the separation of labeled neutrophils from unbound isotope, have been described previously.

Efflu of 45Ca2 from neutrophils Neutrophils were loaded with 45Ca2 for 30 min at 37 C in HBSS which was free of unlabeled Ca2. The cells were then pelleted by centrifuga tion, washed once with, and resuspended in ice cold Ca2 replete HBSS and held on ice until use, which was always within 10 min of completion of loading with 45Ca2. The 45Ca2 loaded neutrophils were then prein cubated for 10 min at 37 C in Ca2 replete HBSS, in the presence and absence of GF10903 , followed by addition of PAF and measurement of the efflu of 45Ca2 over 5 min. The reactions were terminated by the addition of 10 ml ice cold, Ca2 replete HBSS to the tubes which were then transferred to an ice bath. The cells were then pelleted by centrifugation at 400 g for 5 min fol lowed by washing with 15 ml ice cold, Ca2 replete HBSS and the cell pellets finally dissolved in 0.

5 ml of 0. 5% tri ton 100 0. 1 M NaOH and the radioactivity assessed in a liquid scintillation spectrometer. Control, cell free sys tems were included for each e periment and these values were subtracted from the rel evant neutrophil containing systems. These results are presented as the percentage of cell associated radiolabeled cation e truded from the cells. Influ of 45Ca2 into PAF activated neutrophils To measure the net influ of 45Ca2 into PAF activated neutrophils, uncomplicated by concomitant efflu of the radiolabeled cation, the cells were loaded with cold, Ca2 replete HBSS for 30 min at 37 C, after which the cells were pelleted by centrifugation, then washed once with, and resuspended in ice cold Ca2 free HBSS and held on ice until used. Pre loading with cold Ca2 was undertaken to minimize spontaneous uptake of 45Ca2 in Brefeldin_A the influ assay. The Ca2 loaded neu trophils, were then incubated for 10 min in the presence or absence of GF10903 at 37 C in HBSS containing 25 M cold carrier Ca2, fol lowed by simultaneous addition of PAF and 45Ca2 or 45Ca2 only to control, unstimu lated systems.

The 3 tagging plas mids were generated by inserting the PCR product into PstI and BamHI sites of the pCAM BSD hemagglutinin. Transfections were carried out by electroporation of ring stage 3D7 parasites with 75 100 ug selleck chemicals of plasmid DNA, according to Sidhu et al. To select trans formed parasites, 48 h after transfection, Blasticidin was added to a final concentration 2. 5 ug ml. Resistant parasites appeared after 3 4 weeks and were maintained under drug selection. Genotype and phenotype analysis of P. falciparum transfectants To check the presence of correct constructs in transfected parasites, plasmid rescue e periments were carried out. Genomic DNA e tracted from wild or transfected parasites were used to transform E. coli DH5 cells. Plasmid DNA was then purified from bacterial clones and digested with PstI and BamHI.

Genotypes of PfI2 knock out parasites were analyzed by PCR on genomic DNA using standard procedures with the primers Pr 27 and Pr26 specific for the pCAM BSD vector. Genotypes of PfI2 knock in were analyzed using the primer Pr19 and Pr 28. Assays for PfPP1 and effect of PfI2 The activity of PfPP1 with p nitro phenylphosphate was assayed as previously described. To investigate the role of PfI2 recombinant proteins or PfI2 PfI3 derived pep tides on His6 PfPP1 activity, different amounts of proteins were added to 1 ug of PfPP1 recombinant protein and preincubated for 30 min at 37 C before testing the PfPP1 phosphatase activity. Okadaic acid was used as control. Results are presented as mean of increase or de crease of phosphatase activity in comparison to His6 PfPP1 incubated in the reaction buffer.

Yeast two hybrid assays The full length PfPP1 was cloned into the pGBKT7 vector containing the DNA binding domain of gal4 and wild type, deleted or mutated PfI2, PfI2, PfI2W16A, PfI2Y103A into pGADT7 containing the gal4 activation domain. The pGBKT7 Gal4 BD PfPP1 construct was used to transform Y187 strain and maintained on SD media without tryp tophan. The pGADT7 Gal4 AD PfI2 constructs were used to transform AH 109 strain and maintained on SD media lacking leucine. Mating these two haploid strains results in the formation of diploid strain, which is viable on SD media lacking leucine and trypto phan. Interaction of PfPP1 with the different versions of PfI2 proteins were evaluated by their capacity to grow on selective media SD medium lacking leucine, tryptophan and histidine and SD medium lacking leucine, tryptophan, histidine and adenine for 4 days.

Yeasts transformed with empty vector or with pGBKT7 laminine were used as controls. Induction of enopus oocytes germinal vesicle breakdown and co immunoprecipitation Preparation of enopus oocytes and microinjection e periments were performed as previously described. Briefly, AV-951 in each assay, 20 oocytes removed from at least two or three different animals were microinjected with His6 PfI2 recombinant proteins or PfI2 PfI3 derived peptides.

Conclusion In summary, the present results demonstrate that caveo lin 1 e pression is induced after bromocriptine treatment in rat pituitary adenoma cells. Moreover, e ogenous over e pression of caveolin 1 increases apoptosis of pituitary adenoma cells and enhances bromocriptine induced cell apoptosis. Interestingly, sellekchem caveolin 1 was phosphorylated at Tyr14 when GH3 cells responded to bromocriptine treat ment. Phosphorylation of caveolin 1 Tyr14 is predicted to be associated with cellular apoptosis. These data suggest that bromocriptine induced pituitary adenoma cell apop tosis may result from enhanced e pression and activation of caveolin 1 via increased caveolin 1 phosphorylation. Our result e plains the therapeutic effect of bromocriptine in curing pituitary adenoma.

Materials and methods Materials and reagents Dulbeccos modified Eagle medium, penicillin, strepto mycin, L glutamine, fetal bovine serum and horse serum were purchased from Life Technologies. Rabbit anti caveolin 1 and rabbit anti phosphor ylated caveolin 1 antibody were bought from Chemicon Internal Inc. Mouse anti c Myc antibody was obtained from Santa Cruz Bio technology Inc. Te as Red and FITC conjugated secondary antibodies and normal goat serum were purchased from Jackson ImmunoResearch Laborato ries. Plasmid construction and semi quantitated RT PCR Total RNA was e tracted from adult C57Bl 6 mouse brain with TRIzol reagent according to the manufacturers instructions. Caveolin 1 cDNA was amplified from total RNA by RT PCR and was subcloned into pGEM T easy vector by TA cloning.

The DNA sequence of caveolin 1 was confirmed by auto sequencing using an ABI 3730 autosequenser. Caveolin 1 in the pGEM T easy vector was digested with ho I and ba I restriction enzymes and then subcloned into pcDNA4 mammalian e pression vector. this plasmid was termed pcDNA4 caveolin 1. pcDNA4 EGFP was cloned as follows the clone pEGFP N1 plasmid containing enhanced green fluorescent protein, obtained from Clontech, was digested with Pst I and Not I restriction enzymes, then subcloned into pcDNA4 vector. this plasmid was designated pcDNA4 EGFP. pDsRed N1 containing red fluorescent protein was purchased from Clontech Laboratories. For quantifying the level of caveolin 1 cDNA e pressed in GH3 cells modulated after bromocriptine treatment, the cells were treated with either bromocriptine at different concentrations of 5, 10, 20 M or with vehicle for 24 hours, when total RNA was e tracted and the tran scripts quantified by RT PCR, as described above.

Cell culture Dacomitinib The rat GH3 pituitary adenoma and human A431 epithe lial cell lines were obtained from the American Type Cell Collection. The GH3 cells were propagated in F12K nutri ent mi medium supple mented with 2. 5% fetal bovine serum, 15% horse serum, 2 mM L glutamine, 100 units ml penicillin, and 100 units ml streptomycin.

Changes in the zein profile in A69Yo7 seeds were less evident. The o7 mutation decreased the amount of both 22 kDa and 19 kDa a zeins as com pared to wt. However, unlike previously reported, though we did not find clear evidence of a specific polypeptide sup pression mediated by the o7 mutation. Finally, the zein pattern in the o2o7 background was strongly affected, the 22 kDa zein profile was nearly identical to the one observed in the o2, whereas polypeptides of the 19 kDa zein class were decreased both in amount and number. Taken together these data confirm an additive effect of the o2 and o7 mutations in reducing zein accu mulation during endosperm development. Table 1 provides data concerning the percentage con tribution of the main N constituents present in the mature endosperm of the lines considered.

With the exception of non protein N, all N traits measured dif fered significantly, both in amount and composition, between wild type and opaque mutants. The mutant alleles all reduced accumulation of total protein, although to varying extents, the effect being most marked in o7. From these results it was possible to assess the importance of lysine rich non zeins with accuracy, because of the quantification of non protein N and the exhaustive extractions of zeins. Thus, the ratio of non zein content of the endosperm mutants compared with that of the wild type varied from 1. 1 to 2. 2 for the single mutants, whereas for A69Yo2o7, a ratio of 2. 0 was calcu lated. It was also evident that the effect of the o2 is more pronounced in reducing zein accumulation and increas ing the other components than is o7.

This behaviour is also evident in the o2o7 mutant, in which zein synthesis was most reduced, with a concomitant increase in albu min globulins and glutelins, suggesting that in the double t both alleles are active in reducing zein synthesis additively. The overall amino acid compositions of the single mutants o2 and o7, and of the double mutant combina tion o2o7, exhibited a rather similar pattern, although variation was observed of amino acid content in com parison to the wild type endosperms. Each of the single mutants had a high Lys content, whereas o2o7 had more that 3. 5 times the amount present in the wild type. A similar shift, although less pronounced, was observed for Asx, and the other essential amino acids derived from the Asp pathway as well as for Gly, Val, His, and Arg.

AV-951 Among the amino acids reduced in the opaque endosperm mutants were Glx and Leu, the most abundant amino acids found in zein proteins. The reduction of these amino acids gener ally was inversely related to the increase in Lys, with the trend being more evident in the double o2o7 endosperm mutant. Microarray construction Microarray slides were assembled using clones obtained from 20 part normalized cDNA libraries representing the major events in endosperm development.

This is made pos sible by a unique population of adult somatic stem cells called neoblasts. During regeneration and constant homeostatic cell turnover, neoblasts differentiate into all cell types, including germ cells in sexual species. In recent years, several studies have begun to unravel the mechanisms by which regeneration biological activity is regulated at the molecular level. For example, different genes have been shown to play pivotal roles in axon guidance and neuro genesis, the regulation of neoblast proliferation and differentiation, and the re establishment and main tenance of the anteroposterior and dorsoventral body axes. Schmidtea mediterranea and Duge sia japonica are the two planarian species most often used in regeneration studies. There are about 78,000 ESTs for S.

mediterranea in NCBI generated in different projects. Those sequences were clustered to produce a set of 10,000 putative mRNAs which are available from the NCBI Unigene database. The S. mediterranea genome has also been sequenced and assembled at the Genome Sequencing Center at Washington University in St. Louis after approval of a white paper. However, because of this genomes internal com plexity and the lack of a BAC library, its completeness and assembly still needs improvement. A step towards this end was taken when the S. mediter ranea genome and EST information were integrated and approximately 30,000 genes were predicted using an annotation pipeline called MAKER. Those gene models, together with 9,000 mRNAs generated using next generation sequencing technology, were mapped on the planarian genome and used to improve the assembly.

The current assembly contains 43,673 contigs. These are accessible, together with the MAKER annotation data, in the S. mediterranea genome data base. In order to expand our knowledge of the planarian transcriptome and to provide a new tool that can be used to improve the S. mediterranea genome annota tion, we generated a new transcriptome dataset using 454 pyrosequencing technology. The Smed454 dataset can be freely accessed via a website, and the complete sequence data can be downloaded by anyone from there. Mapping of the Smed454 ESTs onto the genome scaffolds shows that the Smed454 dataset con tains more than 3 million nucleotides sequenced de novo. In addition, this mapping extends and connects currently fragmented genomic contigs. Finally, GO annotation of the Smed454 dataset assigns candidate functions to those sequences and facilitates their group ing into distinct gene families. In this way, whole gene families can be analyzed for putative roles Brefeldin_A in planarian regeneration. Thus we are confident that the Smed454 dataset will improve our understanding of how planarian regeneration works at the molecular level.

Tipifarnib clinical Plants have developed various mechanisms to defend themselves against herbivorous insects. In addition to nonspecific, constitutively expressed physical and chemical barriers, plants employ specific induced defenses in re sponse to insect feeding or even egg laying. In contrast to feeding, insect egg laying causes min imal damage to plants, dependent on the egg laying be havior of herbivorous insects, which can be quite distinct in different species. Direct defenses against insect eggs have been reported for crop and herbaceous species including the production of ovicidal substances, growth of neoplasms, development of necrotic zones. Indirect defense against insect egg laying includes induced changes of plant volatile emissions or modifications of the plant surface chemis try attracting or arresting egg parasitoids, which in turn kill the eggs of the herbivores.

The first study demonstrating indirect defense against insect eggs was a study of the field elm, where eggs of the elm leaf beetle induced volatiles which attract the egg parasitoid Oomyzus gallerucae, a tiny eulophid wasp specialized on elm leaf beetle eggs. Elm leaf beetles often feed and lay eggs on the same plant and are known to remove the leaf epidermis prior to egg laying by scratching the leaf surface with their mouthparts. Ex perimental simulation of this egg laying sequence by transferring eggs or oviduct secretion on scratched elm leaves or treatment with jasmonic acid or methyl jasmonate also elicited indirect defense responses in field elms.

A recent study further showed that terpenoids present in the odor of egg induced elm leaves are rele vant for attraction of the egg parasitoids. Induction of attractive plant volatiles by insect egg laying has been shown in one other tree species and two herbaceous crops. The natural range of the European field elm Ulmus minor extends predominantly within South ern Europe. However, through cultivation it occurs throughout the temperate world. Elms are greatly valued for their timber qualities and prior to the Dutch elm dis ease outbreaks, elms were also frequently planted within urban areas because of their environmental tolerance. Many insects including moths, gall mites, and beetles feed on field elms. The elm leaf beetle X. luteola can defoliate entire trees and is recognized as a major urban and forest pest in the USA and Australia. AV-951 The recently published EST sequences for U. americana is to our knowledge, the only other gene expression study of any Ulmus species, where 535 ESTs were identified after trees were exposed to the fungal pathogen Ophios toma novo ulmi, which is the causative agent of Dutch elm disease. Knowledge on how plants are able to respond at the molecular level towards egg laying is scarce.