Escherichia coli from Franklin´s gulls (Leucophaeus pipixcan) at breeding sites in Canada and compared with similar data from the gulls' wintering grounds in Chile. Resistant E. coli phenotypes were common, most notably to ampicillin (30.1%) and cefadroxil (15.1%). Furthermore, 17.0% of the gulls in Canada carried ESBL producing bacteria, which is higher than reported from human datasets from the same country. However, compared to gulls sampled in Chile (30.1%) the prevalence of ESBL was much lower. The dominant ESBL variants in Canada were blaCTX-M-14 and blaCTX-M-15 and differed in proportions to the data from Chile. We hypothesize that the observed differences in ESBL variants are more likely linked to recent exposure to bacteria from anthropogenic sources, suggesting high local dissemination of resistant bacteria both at breeding and non-breeding times rather than a significant trans-hemispheric exchange through migrating birds.Migratory birds have been suggested to contribute to long-distance dispersal of antimicrobial resistant bacteria, but tests of this hypothesis are lacking. In this study we determined resistance profiles and genotypes of ESBL-producing bacteria in randomly selected  Enterobacteriacae with extended-spectrum β-lactamase (ESBL) have increased in frequency in both patients and domestic animals during the last decades, and are now considered as an escalating challenge for global health care . In. InE. coE. coli were isolated from 364 gull samples. These isolates were tested to a panel of 10 antibiotics and 72 (47.4%) isolates were resistant to ≥1 agent, and 16 (9.2%) of the isolates were resistant to ≥3 agents  or CTX-M 9 . Five SHV variants  and one TEM (blaTEM-52) were identified  in the 2009 CANWARD study [Larger differences were seen for the analyses of ESBL-producing bacteria, where the prevalence of ESBLs was lower in Canadian Franklin's gulls (17.0%) than in Chile (30.1%). The ESBL levels in Canadian gulls was, however, significantly higher than the prevalence reported from RD study . SeveralRD study , 26, 29;RD study .blaCTX-M-14 (40.0%) and blaCTX-M-15 (30.6%). In contrast, in the gull samples from Chile, there was a strong dominance of blaCTX-M-1. In the human dataset from Canada, CTX-M dominated among the ESBL and 70.9% of the isolated CTX-M belonged to blaCTX-M-15 [Among the Canadian gull samples CTX-M ESBLs dominated, and the most common genotypes were CTX-M-15 , 26.E. coli were sequence typed, the most common STs were ST 38 and ST 69 (five isolates each), plus six isolates that belonged to the ST10 complex. These STs are also commonly reported from human bacterial collections from Canada, but differed compared to the gull isolates from Chile. In Chile, ESBL isolates showed a greater genetic variation and 31 known STs were detected, including several genotypes that were novel.When E. coli STs do not provide any strong indications of long-distance dissemination between the continents. Generally, gull isolates from Canada had a lower ESBL prevalence rate, dissimilar ESBL genotypes and a more narrow set of E. coli sequence types. In Canada seven among the 11 different STs found in Canadian gulls, were also found in E. coli from Chilean gulls. Thus, the data presented here do not detect any clear signs of frequent long-distance dispersal of E. coli strains, although we acknowledge that sample sizes may have been too low and maybe the influence of the refuse dump in which the Canadian gulls were sampled could have had such a local impact so any signs of trans hemispheric exchange were diluted in this more epidemiological approach. Another plausible hypothesis is that gulls frequently acquire and loose bacteria from the environment which could explain the higher levels of ESBL in wintering gulls where there are a higher local antibiotic dissemination [The comparisons of ESBL genotypes and mination .If persistence of particular resistant strains are short, the population of bacteria sampled may therefore more reflect the local to regional dissemination of resistant bacteria, for example from food-production animals or human waste along the migratory flyways or close to the breeding grounds. Another explanation could be that the pattern seen in Canadian is a result of clonal expansion in the population, resulting in high levels of few genotypes.E. coli can persist in chicken microbiota for more than one month, also in lack of antibiotic pressure [E. coli strains between chickens was observed, suggesting that the population could act as a reservoir. The laboratory experiment was ended after approximately one and a half month, but the carriage pattern indicates that the retention times could be long. In the case of the Franklin's gulls there is no data on how long individual E. coli can colonize its host, and likely the frequent exposure to different isolates as part of the diet may impose competition and/or horizontal gene transfer between strains. The gulls leave their wintering grounds in March-April and reach breeding grounds in late May [In laboratory studies it has been shown that ESBL-producing pressure . Furtherlate May . Thus, aE. coli strains between human activities and gulls in Canada. In both Canada and Chile, gulls were found to carry E. coli ST131, a well-known human-associated strain and the most common ST-type in the human material from Canada [blaCTX-M-14, and not the blaCTX-M-15 which is usually the most widely spread genotype. ST131 with blaCTX-M-14 also occurs regularly in Canadian isolates from humans[The finding of similar STs between Canadian gulls and patients  further E. coli resistant to cefadroxil carried AmpC genes is highly interesting. This could indicate a high AmpC prevalence in the population that should be further studied.The results strengthen the notion that gulls can carry high levels of ESBL and other resistance phenotype and that they thereby could act as a sentinel for dissemination of resistance to the environment. The Canadian gull samples in our study likely received a great part of their bacterial isolates from the refuse dump where they were feeding which illustrates the importance of limiting the possibility for wildlife to be contaminated with human waste products. The levels of ESBL are alarming and that the gulls in this study carry such a high prevalence makes them a possible environmental reservoir of resistant bacterial strains. Although the scoop of this article was classic ESBL, the results showing that all randomly chosen More studies of the ecology, migratory pattern and feeding behavior of birds carrying ESBL-producing bacteria known to be associated with urban regions, are warranted to improve our understanding about whether wildlife play a role in the dissemination of certain antibiotic resistant bacteria.


MicroRNAs (miRNAs) are a class of small noncoding RNAs that can regulate gene expression by binding to target mRNAs and induce translation repression or RNA degradation.There have been many studies indicating that both miRNAs and mRNAs display aberrant expression in breast cancer.Previously, most researches into the molecular mechanism of breast cancer examined miRNA expression patterns and mRNA expression patterns separately.In this study, we systematically analysed miRNA-mRNA paired variations (MMPVs),which are miRNA-mRNA pairs whose pattern of regulation can vary in association with biopathological features, such as the oestrogen receptor (ER),TP53 and human epidermal growth factor receptor 2 (HER2) genes, survival time, and breast cancer subtypes.We demonstrated that the existence of MMPVs is general and widespread but that there is a general unbalance in the distribution of MMPVs among the different biopathological features.Furthermore, based on studying MMPVs that are related to multiple biopathological features, we propose a potential crosstalk mechanism between ER and HER2. MicroRNAs (miRNAs) are a class of naturally occurring small noncoding RNAs. Mature miRNAs are 19- to 25-nucleotide-long molecules that are cleaved from 70- to 100-nucleotide hairpin pre-miRNA precursors . AlthougIncreasing numbers of microRNAs and mRNAs have been found to be related to the development of breast cancer. In contrast to previous studies based only on miRNA or mRNA expression profiles, examining both miRNA and mRNA expression profiles enables us not only to study miRNA and mRNA expression profiles separately but also to examine miRNA-mRNA regulatory pairs together –12. NeveHere, we propose MMPV as a term that indicates miRNA-mRNA pairs whose pattern of regulation can vary in association with different statuses of biopathological features. We reveal that the distribution of MMPVs is widespread. Moreover, we find that the miRNAs of the MMPVs that are associated with a particular biopathological feature tend to display a significant regulatory effect on the target mRNAs related to a specific status of the biopathological feature and tend to display no significant regulatory effect on the target mRNAs related to different statuses. Furthermore, based on studying MMPVs associated with multiple biopathological features, we propose the existence of a potential crosstalk mechanism between ER and HER2. Importantly, this study demonstrates that the pattern of miRNA-mRNA regulation can be altered in the context of different statuses of biopathological features, and this discovery will benefit further research exploring the molecular mechanisms underlying breast cancer.Both miRNA and mRNA expression data were obtained from PMID: 21364938 . The datWe obtained experimentally validated miRNA-mRNA targeting pairs from Tarbase 6.0 . Among tTo examine the regulatory pattern of miRNA-mRNA pairs, which could vary with different statuses of biopathological features, we must quantify the regulatory patterns of the miRNA-mRNA pairs associated with a certain status of a biopathological pattern. For illustrating, here we calculate the regulatory pattern of each miRNA-mRNA pair in ER+ and ER− specimens first. Gene expression with samples in both ER+ and ER− was compiled first, and then the Pearson correlation coefficient (PCC) was adapted to measure the regulatory pattern of miRNA-mRNA pairs associated with a specific status of a biopathological feature. If the PCC is greater than zero, then a positive regulatory pattern corresponds to this PCC and vice versa.q < 0.05).Each miRNA-mRNA pair receives 2 PCCs, corresponding to ER+ and ER− statuses, representing its regulatory pattern in ER+ and ER− specimens, respectively. Based on the quantified results regarding the regulatory pattern of each miRNA-mRNA pair, we prefer those miRNA-mRNA pairs whose 2 PCCs showed opposite algebraic signs (sign change pairs) and those whose 2 PCCs showed the same algebraic sign but displayed the ratio of the PCC of each condition to the normal greater than 2 or smaller than 0.5 (fold change pairs). So that our results would have greater biological importance, we later removed miRNA-mRNA pairs whose 2 PCCs were both insignificant . We used the letters U and D to represent positive and negative regulations and ∗ to indicate the significance in the statistic. Thus, given that the regulatory pattern of experimentally validated miRNA-mRNA pairs downloaded from Tarbase 6.0 in the healthy population was known, we used U or D to represent the regulatory pattern of miRNA-mRNA pairs in breast cancer patients with an ER+ or ER− status and in the healthy population. For example, if the regulatory pattern of hsa-miR-1 and CA3 corresponds to negative regulation in the healthy population and to positive regulation in ER+ breast cancer patients and significant positive regulation in ER− breast cancer patients, then we can refer to this pair as D_U_U*. As another example, the regulation of has-mir-375 and FOLR1 is negative in the healthy population, while in ER+ specimens it is positive, whereas it is significantly negative in ER− specimens. Hence, the change in the pattern of regulation can be represented as D_U_D* to indicate the regulatory pattern of hsa-miR-1 and CA3 in the healthy population and in ER+ breast cancer patients and ER− breast cancer patients.P value threshold is set as 1.0E − 03. The background list comprised all of the genes for the miRNA-mRNA pairs that we obtained from Tarbase 6.0. We placed the mRNAs of each type of MMPV into a target set and obtained the results for the biological process cellular component.GO database was used to explore the biological function involved in MMPVs. We used Gorilla  to conduOur method examines miRNA and mRNA gene expression data to obtain MMVPs for five breast cancer-related biopathological features. The statistical summary is shown in q < 0.05) MMPVs whose miRNA and mRNA are both significantly differentially expressed between the two statuses of the biopathological features (DE-MMPV), and we check the published literature to confirm their relationship with the corresponding biopathological feature. Third, we analyse MMPVs that are shared by multiple types of biopathological features, and we propose the existence of potential crosstalk between ER and HER2. Fourth, mRNAs of each type of MMPV are analysed for GO enrichment through a hypergeometric gene set enrichment analysis. Finally, we map the mRNAs of the MMPVs to Human Protein Reference Database (HPRD) protein-protein interaction networks to explore the topological features of genes of MMPVs.First, we discuss the general unbalanced enrichment trend in the distribution of MMPVs associated with every type of biopathological feature. Second, we select  to evaluate the regulatory effect of miRNAs and discovered that, compared with ER+ patients, most miRNAs exhibit a higher RE score. In other words, they have a more significant regulatory effect on their target mRNAs in ER− patients . This diSuzuki et al. found that three types of missense mutation in the DNA-binding domain of p53 can lead to decreased processing of pri-miRNAs by Drosha . They thMost miRNAs can be regarded as antioncogenesis miRNAs given the fact that compared to the healthy population, most miRNAs are downregulated in cancer patients . MoreoveWe did not find any relevant studies that provide any clues about the unbalance in the distribution of the survival time-associated MMPVs. However, we noted that the regulatory patterns of MMPVs related to ER and HER2 status tended to be significant in ER− and HER2− patients. ER− breast cancer patients are usually resistant to Tamoxifen therapy and, thus, show poorer prognostic results –27. SimiTo explore the relationship between MMPVs and biopathological features, we conducted a significant analysis of microarray (SAM) analysis to detect MMPVs whose miRNAs and mRNAs are both significantly differentially expressed between the two statuses of a given biopathological feature (DE-MMPVs). The final results are shown in First, we analysed the differentially expressed genes (DEGs) among ER-associated DE-MMPVs, and we found that they shared the same miRNA: hsa-miR-375. Pedro de Souza Rocha Simonini reported that hsa-miR-375 is overexpressed in breast cancer tumours with an ER+ status and that decreasing the expression of hsa-miR-375 will decrease the activity of ER accordingly . This obSecond, we analysed the DEGs among TP53-associated DE-MMPVs. Adan Valladares showed that CCND1 and LAMC1 are overexpressed in breast cancer patients . We obseIt can be observed that the regulatory pattern of hsa-let-7b and CCND1 remains positive in breast cancer patients, regardless of the status of TP53, which is negative in the healthy population. Such D_U_U regulatory pattern variation was also found for hsa-miR-145 and MUC1. MUC1 encodes a mucoprotein that is overexpressed in many types of cancer, including breast cancer. Similar to CCND1, MUC1 is a potential biomarker for tumours, and MUC1 plays an important role in the invasion and metastasis of tumours. Specifically, MUC1 can interact with TP53 and inhibit cell apoptosis mediated by TP53, thus facilitating the proliferation of cancer cells . SimilarNguyen et al. reported that hsa-miR-29c negatively regulates DNMT3B, the overexpression of which can cause the hypermethylation of some tumour suppressor genes . Our resFinally, we analysed the DEGs among the subtype-associated DE-MMPVs. We observed that 2 MMPVs  and one mRNA (CDK6) are shared by the TP53-associated MMPVs and subtype-related MMPVs. Specifically, compared to patients with mutated TP53, the levels of MUC1, CBFB, and CDK6 expression are significantly decreased, significantly increased, and significantly decreased, respectively, in wild type TP53 patients. Interestingly, compared to basal-like breast cancer patients, the levels of MUC1, CBFB, and CDK6 expression are significantly decreased, significantly increased, and significantly decreased in nonbasal-like breast cancer patients, respectively. All of these findings indicate that these three genes could play similar roles in TP53 pathways and biological pathways that are related to basal-like breast cancer. Moreover, we found that, compared to basal-like breast cancer patients, the expression of ETS is significantly decreased in nonbasal-like cancer patients. This finding is supported by the work of Charafe-Jauffret et al., who reported that the expression of ETS1 is higher in basal-like breast cancer than in other breast cancer subtypes . FurtherWe found that many MMPVs are associated with multiple biopathological features. The distribution of these MMPVs is shown in To find genes that are closely associated with two different biopathological features, we further filtered the data that appear in β1 and, hence, enhance the growth of cancer cells [β1 can repress the expression of MYB, and ER+ status enhances the expression of MYB [There is a substantial amount of research examining crosstalk between ER and HER2. Isabel Pinhel claimed that the levels of HER2 and ER expression are positively correlated in non-HER2-overexpressing breast cancer tumours and are negatively correlated in HER2-overexpressing breast cancer tumours . HER2 over cells . Moreoven of MYB . Importan of MYB . An expen of MYB . Furthern of MYB . ConsideThis proposed potential crosstalk between MYB, overexpressed HER2, and ER not only will contribute to further studies addressing the molecular mechanisms underlying breast cancer but also serves as an important reference for potential joint therapy with tamoxifen and trastuzumab.http://dx.doi.org/10.1155/2014/291280.We conducted a GO enrichment analysis of the mRNA components of MMPVs. The results of the biological process (BP) enrichment analysis and cellular component (CC) enrichment analysis are shown in Supplementary Table 1 in Supplementary Material available online at It can be observed that the GO terms related to TP53 mainly include molecules that are involved with cell adhesion to the extracellular matrix. There have been many studies concentrating on the relationship between TP53 and tumour metastasis, which is closely associated with cell adhesion and the extracellular matrix. Specifically, Abramson et al. reported that compared to individuals with wild type TP53, the strength of cell adhesion is greatly increased in the population exhibiting mutant TP53 . AnagantThe GO terms related to the ER are mainly associated with DNA synthesis and the cell cycle. S F Doisneau-Sixou claimed that oestrogen independently regulates the expression and function of c-Myc and cyclin D1. Antioestrogen treatment of MCF-7 cells can greatly decrease the expression of c-Myc and cyclin D1, resulting in the arrest of the cell cycle and inhibition of DNA synthesis.t-test to compare the average degree of the MMPV genes with those of PPI network. In fact, there are biases which existed in current PPI databases. Here we use the commonly used database, HPRD. The results are shown in We explored the topological features of the genes encoding MMPVs by mapping the mRNAs of each type of MMPV to the HPRD protein-protein-interaction (PPI) network . SpecifiP values were calculated using a one-tailed t-test. The P value shown in bold indicates that the average degree of the corresponding biopathological feature is significantly greater than that in the HPRD-PPI network.In this study, we discovered that the regulatory pattern of miRNA-mRNA pairs can vary with different statuses of biopathological features. To further explore the molecular mechanisms underlying breast cancer, we studied five biopathological features  that are closely related to breast cancer. We observed a general unbalance in the distribution of MMPVs. Moreover, the differentially expressed MMPV genes suggest that there is a potential effect of these biopathological features on the development of breast cancer at the molecular level. Furthermore, we examined the topological features of genes encoding MMPVs in the HPRD PPI network, and we propose the existence of potential crosstalk between ER and HER2. The method developed in this paper can help detecting biologically important regulatory events mediated by miRNAs.Gene ontology (GO) enrichment analysis was done on cellular component and biological processes branches. GO terms related to TP53 mainly include molecule that are involved with cell adhesion to the extracellular matrix.Click here for additional data file.


MAP4 gene region and on chromosome 3 with blood pressure. To prioritize variants across the genome for testing, variants were first functionally assessed using prediction algorithms and expression quantitative trait loci (eQTLs) data. Four set-based tests in the family-based association tests (FBAT) framework--FBAT-v, FBAT-lmm, FBAT-m, and FBAT-l--were used to analyze 20 pedigrees, and 2 variance component tests, sequence kernel association test (SKAT) and genome-wide complex trait analysis (GCTA), were used with 142 unrelated individuals in the sample. Both set-based and variance-component-based tests had high power and an adequate type I error rate. Of the various FBATs, FBAT-l demonstrated superior performance, indicating the potential for it to be used in rare-variant analysis. The updated FBAT package is available at: http://www.hsph.harvard.edu/fbat/.The revolution in next-generation sequencing has made obtaining both common and rare high-quality sequence variants across the entire genome feasible. Because researchers are now faced with the analytical challenges of handling a massive amount of genetic variant information from sequencing studies, numerous methods have been developed to assess the impact of both common and rare variants on disease traits. In this report, whole genome sequencing data from Genetic Analysis Workshop 18 was used to compare the power of several methods, considering both family-based and population-based designs, to detect association with variants in the  Both existing and novel methods incorporating family-based and population-based designs were compared in this report. All the methods we compare use a single test for a set of multiple single-nucleotide polymorphisms (SNPs) in a region (gene in our setting). This approach avoids the problem of needing large samples for testing rare variants individually.family-based association tests (FBAT) refers to a suite of family-based association testing methods that rely on an extension of the transmission disequilibrium test. We used 2 newly developed rare-variant association tests in the framework of FBAT, FBAT-v, FBAT-lmm, and 2 previously existing multimarker FBAT tests, FBAT-m and FBAT-l. Although the Genetic Analysis Workshop 18 (GAW18) sample size is large, it is made up of a small number of pedigrees with a large number of individuals per pedigree. The FBAT approach treats all nuclear families in a pedigree as independent, unless a trait locus is known to be linked to the markers under test. The Q1 variable, which was not simulated to be directly associated with any causal gene, was very highly heritable  was used to predict nonsynonymous, splice, and stop variants. Nonsynonymous variants were further classified using polyphen2 [MAP4 [10 p value from eQTL analysis) was used for eQTL filtering.Variants were filtered based on their predicted function. For coding variants, SnpEff  has been proposed as a test for association between both common and rare genetic variants in a region using either continuous or dichotomous traits ,7 for poWe used the complete set of 200 replicates for assessing type I error and power, using an alpha of 0.05 to determine statistical significance. In our analyses, we focused on 2 continuous phenotypes: systolic blood pressure (SBP) and diastolic blood pressure (DBP). Heritability estimates for SBP and DBP were both in the range of 20% to 30% . In total, 1443 genes were analyzed for their association with average residual of blood pressure over three time points  overlap with the results from SKAT.A whole genome scan was performed using FBAT-l for both family and population-based methods after adjusting for first 10 principal components generated by EIGENSTRAT . Only chMAP4 gene and on chromosome 3. This approach incorporated the use of functional prediction information to filter variants as would traditionally be done in most applied studies. Both SKAT and GCTA had high power and an adequate type I error rate. Of the various FBAT tests, FBAT-l demonstrated superior performance, indicating the potential to be used in rare-variant analysis. The lack of population substructure and availability of potential phenotypes contribute to the high performance of FBAT-l. Absent these conditions, the performance degrades. The relatively poor performance of FBAT-lmm could be a result of small sample size and concordant direction of effect size across SNPs. However, FBAT-lmm shows promise for the case where effect sizes within a test region vary in signs of risk. It does not currently have the capability to analyze extended pedigrees.Both family-based and population-based analyses of whole genome sequencing data were evaluated for their power to detect associations with a simulated phenotype with variants in the We also note that when analyzing extended pedigree data and highly correlated traits between relatives, the empirical variance estimator (-e) should be used to achieve the correct type I error. However, its use decreases the effective sample size so that it is closer to the number of independent pedigrees. Finally, our analysis demonstrates that using the average phenotype over 3 time points gives higher power compared to single-time-point phenotype analysis. This suggests the combination of the phenotypes from different time points, or even the combination of SBP and DBP, may achieve higher power.MAP4 and this could be due to some causal variants of MAP4 within the variants for analysis being common. Our population-based tests comparison suggests that in the absence of population substructure, the population-based association tests are more powerful.In this paper, we compared various FBAT region based tests and compared family based tests with population based tests. Our results show that FBAT -l outperformed FBAT -v0 when testing The authors declare that they have no competing interests.JJZ developed FBAT-lmm, ran analyses, and wrote the paper. WKY codeveloped FBAT-v and FBAT-v -e, and assisted in computing. MHC developed the functional screening algorithm for SNP reduction. DQ assisted in data cleaning and formatting, and running of principal components. MLNM edited and revised the manuscript; NML performed the overall design and edited the manuscript. All authors read and approved the final manuscript.


Ulcerative colitis (UC) is a major form of inflammatory bowel disease (IBD) worldwide. Better understanding of the pathogenesis of UC has led to the development of novel therapeutic agents that target specific mediators of the inflammatory cascade. A number of biological agents have been approved by the U.S. Food and Drug Administration (FDA) for the treatment of UC and several more are currently in various phases of drug development. The commonly used agents include TNFα antagonists  and anti-integrin agents (vedolizumab). These biological agents have profoundly influenced the management of UC patients, especially those with refractory disease. This paper reviews the currently available knowledge and evidence for the use of various biological agents in the treatment of UC. Ulcerative colitis (UC) is a major form of inflammatory bowel disease (IBD), characterized by chronic inflammation involving the colon and rectum. It is a cause of significant morbidity worldwide and its incidence and prevalence appear to be increasing with time . PatientAmong the pro-inflammatory cytokines, the role of TNF-α has been the most extensively studied. Excessive production of TNF-α from activated macrophages and T-lymphocytes leads to further activation of macrophages and T-lymphocytes, expression of adhesion molecules on vascular endothelium and recruitment of neutrophils, resulting in a vicious cycle of increasing inflammation , 5, 6. EDespite promising results with the use of anti-TNF agents in the management of UC, some patients continue to require corticosteroids or develop primary or secondary non-response to anti-TNF therapy. Failure to respond to biological therapy is categorized into two types: primary and secondary failure. In patients with the primary failure, no improvement in clinical signs and symptoms is noted with initial induction therapy using the treatment agent. In contrast, patients with the secondary failure exhibit initial improvement with induction therapy and subsequent maintenance therapy but eventually lose the response. As a result, several other ‘key players' of inflammatory pathways have been investigated as therapeutic targets. These include adhesion molecules, which help in migration of leukocytes , other cytokines  and T-lymphocyte receptors (such as CD28 and CD25).This review provides updated information on various biological agents in the treatment of UC.The widespread use of anti-TNF-α agents has changed the treatment paradigm in the management of patients with UC, as well as in those with CD. Several strategies for targeting and inhibiting TNF-α have been developed and include neutralization of TNF-α with monoclonal antibodies and inhibition of its production or its receptors . Four agInfliximab was the first agent to receive FDA approval for induction and maintenance therapy in UC . It is aThe efficacy of infliximab in the treatment of UC has been demonstrated in both open-label and randomized, controlled trials . Two larThe efficacy of infliximab increases further when used in combination with azathioprine. In a recent double-blinded, placebo-controlled trial, 239 anti-TNF-α therapy-naïve patients with moderate-to-severe UC were randomized to receive either (i) intravenous infusions of infliximab (5 mg/kg dose) at weeks 0, 2, 6, and 14, plus daily oral placebo capsules or (ii) oral azathioprine (2.5 mg/kg daily) plus placebo infusions on the infliximab schedule or (iii) combination therapy with both drugs . SignifiA meta-analysis of five randomized, controlled trials, examining the efficacy of infliximab in the treatment of moderate-to-severe UC, showed that infliximab therapy was associated with a higher rate of induction of remission than placebo .Infliximab has also been used for the treatment of chronic pouchitis, which is a common complication after proctocolectomy with ileal pouch anal anastomosis surgery for UC; however, experience with the use of infliximab in this setting is currently limited to case reports and series . AlthougAdalimumab is a fully humanized monoclonal antibody against TNF-α which is administered subcutaneously. It was approved by the FDA in 2012 for the treatment of moderate-to-severe UC in adults after its efficacy was demonstrated in several randomized and non-randomized studies . BecauseTwo large, randomized, controlled trials demonstrated the efficacy of adalimumab for the induction and maintenance of remission in UC patients. These were the UC Long-Term Remission and Maintenance with Adalimumab 1 and -2 trials  , 38.vs. 9.2%; P = 0.031). There was no significant difference in remission rates between patients receiving adalimumab 80/40 and placebo . Simultaneously, patients receiving adalimumab 160/80 showed significantly greater improvements in nutrition and inflammatory markers than the placebo group.ULTRA 1 was an 8-week, multicenter, randomized, double-blinded, placebo-controlled trial investigating the use of adalimumab as induction therapy in patients with moderate-to-severe UC despite conventional therapy . In thisThis study was followed by ULTRA 2, which was a 52-week study assessing the efficacy of adalimumab as maintenance therapy in UC patients . This trAn open-label extension study to ULTRA 1 and 2 is currently under way, to assess the long-term efficacy of adalimumab in UC patients . PrelimiGolimumab is a fully humanized monoclonal antibody against TNF-α and is the latest anti-TNF agent to receive FDA approval for treatment of moderate-to-severe ulcerative colitis. Similarly to adalimumab, it is administered subcutaneously and can therefore be self-administered by patients at home. Data regarding the efficacy of golimumab in UC comes mainly from two large, double-blinded, randomized, controlled trials, the Program of Ulcerative Colitis Research Studies Utilizing an Investigational Treatment, which was divided into Subcutaneous and Maintenance phases  , 41. ThePURSUIT-SC was an integrated phase-II and phase-III trial, in which 1064 anti-TNF therapy-naïve patients with moderate-to-severe UC  despite conventional therapy were included . Of thesThe subsequent PURSUIT-M trial included the same participants as PURSUIT-SC, to study the efficacy of golimumab as maintenance therapy . In PURSin vitro [Certolizumab pegol is the first pegylated, humanized monoclonal antibody against TNF-α. In fact, it contains only the antigen binding fragment (Fab) of the monoclonal antibody which has been conjugated to polyethylene glycol and does not contain the crystallizable (Fc) or tail portion of the antibody . These uin vitro . Lack ofin vitro .Certolizumab pegol has been shown to be effective in the treatment of CD and was approved by the FDA for this indication , 47; howIntegrins are a group of transmembrane receptors, which are involved in cellular adhesion and signal transduction. Integrins play an important role in the inflammatory response by participating in the adhesion of leukocytes to vascular endothelium and allowing it to migrate across the blood vessels at the inflammatory site . BlockinVedolizumab is a humanized monoclonal antibody that selectively binds to α4β7 integrin, which is located exclusively on the surface of gut-homing leukocytes . By bindThe efficacy of vedolizumab in the treatment of UC has been demonstrated in the GEMINI 1 trial . This waFor the maintenance trial, patients from either above group who had a response to vedolizumab were randomized to receive either vedolizumab or placebo every 4 or 8 weeks. At the end of 52 weeks, patients receiving vedolizumab were significantly more likely to have clinical remission and clinical response than with placebo, including those who had previously failed anti-TNF therapy. Additionally, patients in the treatment group were less likely to require corticosteroids and had greater improvements in quality of life scores than those in the placebo group. There was no significant difference in efficacy between the two vedolizumab regimens. Also, the rate of adverse events was similar in the treatment and placebo groups and no cases of PML were observed during the 52 week follow-up.Vedolizumab was recently approved by the FDA for the treatment of adults with moderate-to-severe UC . An openDespite the recent advances, many patients fail to respond to medical therapy with currently available agents. Ongoing research on the pathogenesis of IBD has provided several more attractive therapeutic targets, for which new drugs are currently under development.PF-547659 is an investigational monoclonal antibody against MAdCAM-1 (addressin), which is the extracellular ligand for α4β7 integrin. It inhibits adhesion and trans-endothelial migration of leukocytes by suppressing the interaction of leukocytes with vascular endothelium, similarly to vedolizumab. Favorable results were observed in a preliminary study in UC patients and further trials are in progress .Alicaforsen is another anti-adhesion molecule that reduces lymphocyte migration. It is an anti-sense oligonucleotide, causing decreased synthesis of endothelial adhesion molecule ICAM-1. A recent meta-analysis of four phase II studies in UC suggests that alicaforsen is effective for the treatment of moderate-to-severe UC, especially in patients with distal disease . In an oAlthough the main focus of treatment of patients with UC has traditionally been the alleviation of symptoms by inducing and maintaining symptomatic remission, there is increasing evidence to suggest that achieving mucosal healing and reduction in endoscopic disease activity may be as critical as improvement in symptoms in optimizing long-term outcomes . Indeed,P < 0.009 for all comparisons). Similarly, during the ULTRA2 trial, which assessed the efficacy of adalimumab, mucosal healing rates were higher in the treatment group than those in the placebo group both at week 8 (P = 0.032) and week 52 (P = 0.009) [The efficacy of biological agents in reversing the tissue damage associated with inflammation has been well studied. The efficacy of infliximab in mucosal healing was assessed during the ACT1 and ACT2 trials . Mucosal= 0.009) . Higher = 0.009) , 41.Mucosal healing effects have also been demonstrated with the use of vedolizumab. In the GEMINI 1 trial, patients treated with vedolizumab experienced higher rates of mucosal healing than placebo at both week 6 and week 52 [The natural history of UC is that of a progressive disease leading to high rates of surgery and morbidity . TreatmeUsing data from the ACT1 and ACT2 trials, a subsequent study demonstrated that patients treated with infliximab had a significantly lower rate of colectomy by week 54 than those treated with placebo . AdditioIn another randomized, double-blinded, placebo-controlled trial of 45 hospitalized patients with intravenous steroid-refractory flare of moderate-to-severe UC, patients who received a single infusion of infliximab after administration of steroids required colectomy significantly less frequently at 3 months following the infusion, than those who received placebo . A folloSimilar results have been demonstrated with use of adalimumab. Using data from the ULTRA 2 trial, subsequent studies have indicate that adalimumab therapy is associated with significantly reduced risk of hospitalization and significantly greater improvement in health-related quality of life measures during 52 weeks than with placebo , 66.The randomized, controlled trials have demonstrated a generally favorable safety profile for biological agents; however, a few patients do experience side-effects and biological agents have been occasionally associated with severe—sometimes life threatening—adverse effects necessitating careful monitoring of therapy . Many ofPatients receiving biological therapies are at an increased risk of opportunistic infections and re-activation of latent infections, such as tuberculosis and hepatitis B virus. This risk is further increased by combination therapy with other immunomodulators . As mentBiological agents are also associated with an increased risk of malignancies, especially lymphoma . This isMany of the biological agents are immunogenic and patients frequently develop antibodies against these drugs, which can interfere with their therapeutic effects. This problem is more frequent with chimeric agents like infliximab, which are considered more immunogenic than with fully humanized agents like adalimumab . Also, pA number of biological agents are currently available for the treatment of patients with IBD . These aEd and Joey Story Endowed Chair.The research and education activity of Bo Shen is supported by the Conflict of interest statement: none declared.


Fear of eye gaze and avoidance of eye contact are core features of social anxiety disorders (SAD). To measure self-reported fear and avoidance of eye gaze, the Gaze Anxiety Rating Scale (GARS) has been developed and validated in recent years in its English version. The main objectives of the present study were to psychometrically evaluate the German translation of the GARS concerning its reliability, factorial structure, and validity.Three samples of participants were enrolled in the study. (1) A non-patient sample (n = 353) completed the GARS and a set of trait questionnaires to assess internal consistency, test-retest reliability, factorial structure, and concurrent and divergent validity. (2) A sample of patients with SAD (n = 33) was compared to a healthy control group (n = 30) regarding their scores on the GARS and the trait measures.The German GARS fear and avoidance scales exhibited excellent internal consistency and high stability over 2 and 4 months, as did the original version. The English version’s factorial structure was replicated, yielding two categories of situations: (1) everyday situations and (2) situations involving high evaluative threat. GARS fear and avoidance displayed convergent validity with trait measures of social anxiety and were markedly higher in patients with GSAD than in healthy controls. Fear and avoidance of eye contact in situations involving high levels of evaluative threat related more closely to social anxiety than to gaze anxiety in everyday situations.The German version of the GARS has demonstrated reliability and validity similar to the original version, and is thus well suited to capture fear and avoidance of eye contact in different social situations as a valid self-report measure of social anxiety and related disorders in the social domain for use in both clinical practice and research. The face, especially the eye region, provides a multitude of social information . Eye conFear and avoidance of eye-to-eye contact is a frequently reported clinical symptom of Social Anxiety Disorder (SAD). One explanation for reduced eye gaze in SAD relates to the need to avoid a possible source of threat, as the eyes convey socially evaluative information . From anIn using systematic observer ratings, some studies indeed reported inadequate eye contact in SAD compared to healthy controls –11. The Despite this growing evidence for reduced eye gaze in SAD relying on objective experimental data, less is known about the subjective experience of eye contact in SAD. Schneier et al.  recentlyWe aimed in this study to assess the psychometric properties of the German translation of the GARS , to replicate the factorial structure of the GARS, and to further assess self-reported fear and avoidance of eye gaze in a sample of socially anxious individuals and matched healthy controls.The first sample of participants for evaluating reliability and factorial structure of the GARS comprised n = 353 adults recruited via local on-campus advertising. This sample mainly comprised students , and employees from different professions  aged a mean 29.4 years . About two-thirds of the sample were women  and most had higher education . Psychology students were excluded, as were participants reporting a lifetime history of mental illness or current severe somatic illness. To determine the test-retest reliability, two non-overlapping subsamples of this group joined the second enrollment 2 months (n = 76) and 4 months (n = 73) after the first survey.In all, 1208 persons were contacted by e-mail, of whom 436 responded by starting the survey. Of these, 359 completed the first evaluation questionnaire yielding 353 valid data sets for reliability analysis, and 328 data sets for correlational analyses. From the initial sample, a random subsample of 156 persons was contacted by e-mail at the first retest interval (2 months), 105 responded, and 76 completed the second survey. For the second retest interval (4 months), we contacted another subsample of 156 persons via e-mail: 90 responded, 73 completed the second survey.To validate the GARS, we enrolled a group of n = 33 male participants with GSAD from an ongoing study unrelated to the present evaluation. The GSAD diagnosis was confirmed via the Structured Clinical Interview for DSM-IV (SKID ). GSAD pThe procedures in this study were approved by the local ethics committee of the University of Freiburg. All participants gave written informed consent before participation and received monetary compensation.All participants completed the GARS, and a battery of questionnaires comprising questionnaires on demographic variables, trait anxiety, social anxiety, and personality traits within approx. 30 minutes.12 = .99 [The GARS was originally developed from unstructured interviews with GSAD patients and comprises 17 situations involving enhanced fear or avoidance of eye contact reported by these patients . The oriIn addition to the GARS, participants completed the Social Interaction Anxiety Scale (SIAS ), a wideThe Brief Fear of Negative Evaluation—revised (BFNE-R) is a 12-item self-report questionnaire measuring fear and distress related to negative evaluation by others on 4-point Likert scales The revised brief version is based on the original Fear of Negative Evaluation Scale  and has General trait anxiety was assessed via the trait scale of the State-Trait Anxiety Inventory (STAI-t ). This wDepressive symptoms were assessed with the German version of the Center of Epidemiological Studies Depression Scale (CES-D ). The shFinally, a 30-item version of the Neo-Five-Factor-Inventory (NEOFFI ) was empA subsample of the initial group was re-assessed via the GARS two months after the first assessment, and another independent subsample completed the GARS four months a second time. All questionnaires were presented with EFS survey 10.6 using secure connection via the internet . Each participant was sent a single, unique keycode by e-mail to access the online survey and complete the questionnaires at home.Reliability was estimated by calculating internal consistencies (Cronbach’s α) for the subscales and total score of the GARS. In addition, item-total correlations were calculated and split-half-reliabilities using the odd-even-methods with Spearman-Brown correction. Test-retest reliabilities were estimated by calculating intraclass correlation coefficient (ICC) and Pearson-correlations for two retest intervals: at 2 and 4 months.The factorial structure was determined by calculating two independent principal component analysis (PCA) with a Varimax rotation for GARS-fear and GARS-avoidance ratings separately. A confirmatory approach with a two-factor solution was chosen to replicate the preliminary findings with the original English version ,18.Convergent and discriminant validity was investigated by calculating the bivariate correlation coefficient of the GARS subscales with the SIAS, BFNE, STAI-T, CES-D and the subscales of the NEOFFI-30. We also explored the predictive power of the two factors using correlation analyses.t-tests were calculated for the fear and avoidance subscales, the total score and the two factors revealed in the PCA.Finally, we tested criterion-related validity by comparing the GARS subscales in a group of participants with SAD to a healthy control group. For this purpose, p<.05. Correction for multiple testing was achieved by correcting the exact p-values according to Bonferroni.All analyses were conducted with SPSS for Windows (Version 22). Significance threshold was Descriptive data of the item-analysis are illustrated in Calculation of GARS-fear and GARS-avoidance sub-scores resulted in comparable mean values . The correlation between GARS-fear and GARS-avoidance sub-scores was r = .78 (p<.001). Age was not associated with the GARS-fear score , but there was a small yet significant negative correlation for the GARS-avoidance . Educational level was not associated with the GARS scales . Women revealed slightly higher scores on both subscales and the total score .Internal consistencies (Cronbach’s α) for the subscales and the total score were very high, ranging from .90 to .95. Accordingly, split-half reliabilities of the subscores and total scores was very high, ranging from r = .82 to r = .86 for uncorrected, and from r = .91 to r = .93 for Spearman-Brown corrected correlations. Finally, test-retest correlations over an interval of 2 and 4 months revealed high to medium stability of the test scores in two independent samples see .The confirmatory factor analysis for the GARS-fear ratings with Varimax rotation revealed two distinct factors with high loadings (r>.30) of 14 items. Three items showed substantial loadings on both factors. Explained variance of the rotated solution was 52.8 percent. In the GARS-avoidance ratings, 13 of the 17 items showed distinct loadings (r>.30) on one of the two factors. The two-factor solution explained 46.4 percent of the variance observed  everyday situations  with 6 items, (2) situations involving high levels social threat  with 8 items. Subscores calculated by summing up the items on each factor correlated positively for the fear ratings  and avoidance ratings .Exploring sex differences in fear of eye gaze in the different situational factors using a 2-way ANOVA, we noted a significant sex-by-situation interaction . Post-hoc tests revealed significantly higher levels of fear in women in conjunction with social-threat situations , but not in everyday situations . We observed the same pattern in the avoidance ratings .Correlation analyses revealed positive correlations with trait anxiety, depression, fear of negative evaluation, and social anxiety for both the fear and avoidance sub-scores of the GARS. Unsurprisingly, we observed the highest correlation for social anxiety, confirming the diagnostic relevance of reduced eye contact in social phobia see . Divergepart = .56. Using the situational factors for correlation analyses, we found higher correlations for situations of social threat than everyday situations. In particular, fear and avoidance of eye gaze in socially-threatening situations (factor 2) were the better predictors for social anxiety (r = .63 and r = .61) than fear and avoidance in everyday situations .The association between the total GARS score and social anxiety (SIAS) remained significant after controlling for trait anxiety (STAI-t): GARS-total/SIAS rGroup differences between GASD patients and matched healthy controls were calculated for the GARS subscales and the two situational factors see . OverallAs expected, the GSAD patients displayed markedly enhanced fear and avoidance in both everyday (factor 1) and social threat situations (factor 2). However, a three-way ANOVA with group (HC vs. GSAD), scale (fear vs. avoidance), and situation (everyday vs. threat) revealed a significant group x situation interaction , indicating a stronger tendency to report high levels of fear and avoidance of eye gaze in threatening situations in GSAD, as compared to healthy controls. Calculation of effect sizes (Cohen’s d) confirmed the greater difference in gaze fear and avoidance in GSAD patients in social threat situations (factor 2).Exploratory analysis of the descriptive data in the additional GARS items revealed consistently higher scores on the anxiety-associated attributions of gaze avoidance in patients with GSAD see .th birthday (77%). Regarding how their difficulties developed from childhood to adulthood (item 25: “My current anxiety and avoidance related to eye contact is… (0) worse than in my childhood … (3) a lot better than in my childhood”), 39% of the GSAD patients reported worsened, 26% unchanged, and 35% less gaze anxiety.Interestingly, GSAD were less convinced that making eye contact is important for their social interactions (item 23). Participants with GSAD reported a large range (4 to 45 years) concerning the age of onset of their problems making eye contact (item 24 “Earliest age when I felt anxious about eye contact or avoiding eye contact”), but most reported that they started having difficulty before their 18The present study investigated the psychometric properties of the German version of the GARS in a sample of non-patients and in patients with GSAD. Our data provides evidence of excellent internal consistency and high reliability over several months. The factorial structure of the original English version was replicated with the German version and reveals the two distinct “everyday situations” vs. “situations with evaluative threat”-factors. We noted a strong linear association between fear and avoidance of eye contact with social anxiety in non-patients. Accordingly, fear and avoidance of eye contact was higher in patients with GSAD than in healthy controls. Overall, the psychometric properties of the German version of the GARS verify the usefulness of this self-report questionnaire to measure the subjective experience of eye contact in various situations.Our assessment of internal consistency and test-retest reliability revealed that the German GARS is as reliable as the original version . The staThe factorial structure of the GARS in the present study concurs overall with previous reports . For botAs expected, GARS scores’ concurrent correlations with well-established measures of social anxiety (SIAS), general anxiety (STAI-t), fear of negative social evaluation (BFNE), and depression (CES-D) were all positive and lower to medium range. In contrast, high levels of fear and avoidance of eye contact predicted lower levels of extraversion and conscientiousness. The maximum explained variance was about 40% for social anxiety, and about 10% for extraversion, showing that self-reported fear and avoidance of eye gaze has a limited predictive power for complex personality traits. However, after controlling for general anxiety, total GARS scores still explained about 30% percent of the variance in social anxiety, suggesting a specific amount of common variance between difficulties in eye gaze and social anxiety. Notably, the association between fear of negative evaluation and fear and avoidance of eye gaze was modest, with 13% shared variance.Women reported slightly higher levels of fear and avoidance of eye gaze in this study, a finding consistent with the higher prevalence of social anxiety in women in the general population . InteresAs expected, group comparison between the GSAD patients and our age-matched control group revealed markedly higher levels of fear and avoidance in social anxiety. This effect was pronounced in conjunction with situations involving high levels of social threat (factor 2). The overall scores the GSAD group obtained in the present study were somewhat lower than those of the sample that Schneier et al. examined , which iIt should be noted that the GARS is based on self-reporting and thus behavioral validation; more objective measures of eye contact in social situations using appropriate techniques (e.g. eye-tracking) are still needed. Furthermore, since the present study investigated only male patients with GSAD, potential sex-specific differences in fear and avoidance of eye contact in GSAD should be specifically addressed in future studies. The specificity of gaze fear and avoidance needs to be investigated by comparing patients suffering from social anxiety with patients presenting other mental disorders involving severe social deficits, such as anxiety disorders, autism spectrum disorder, schizophrenia, or chronic depression. Finally, cultural background is a significant modulator in non-verbal communication behavior, exerting a particular influence on eye gaze behavior and the subjective response to direct eye contact during conversations . Given tThe German version of the GARS is a reliable and valid instrument with which to assess self-reported fear and avoidance of eye contact in various social situations. The two-factor structure of the original questionnaire has been replicated in the German version, indicating that fear and avoidance of eye contact can be differentiated in two categories of social situations: (1) everyday situations and (2) those involving high social threat. The GARS is well suited to assess the dispositional self-reported tendency to fear and avoid eye contact in social situations and might thus be useful as a sensitive measure for social anxiety in different clinical conditions such as SAD and other disorders associated with social impairments such as autism or schizophrenia.S1 Appendix(PDF)Click here for additional data file.


At the macroscopic level, similar effects can be induced in the human brain using repetitive stimulation with identical stimuli. High-frequency stimulation (HFS) can increase neuronal responses whereas low-frequency stimulation may produce the opposite effect. Optimal stimulation frequencies and characteristics for inducing stimulus-specific response modification (SRM) differ substantially from those applied to brain tissue slices but have been explored in recent studies. In contrast, the individual manifestation of this effect in terms of its spatial location and extent are unclear. Using functional magnetic resonance imaging (fMRI) in 18 subjects (mean age 25.3 years), we attempted to induce LTP-like effects by HFS with checkerboard flashes at 9 Hz for 120 s. As expected, flashes induced strong activation in primary and secondary visual cortices. Contrary to our expectations, we found clusters of decreased activations induced by pattern flashes after HFS in the primary and secondary visual cortices. On the level of the individual subject, some showed significantly increased activations in the post-HFS session while the majority showed significant decreases. The locations of areas showing altered activations before and after HFS were only partly overlapping. No association between location, extent and direction of the HFS-effect was observed. The findings are unexpected in the light of existing HFS-studies, but mirror the high inter-subject variability, concerning even the directionality of the induced effects shown for other indices of LTP-like plasticity in the human brain. As this variability is not observed in LTP at the cellular level, a better understanding of LTP-like mechanisms on the macroscopic level is essential for establishing tools to quantify individual synaptic plasticity  The brain's ability to adapt is referred to as neuronal plasticity. Neuronal plasticity is important in the context of learning and reorganization of the brain when facing acute  or continuous damage .In vitro, tetanic stimulation at relatively high frequencies (>30 Hz) induces LTP, whereas trains of stimulation at low frequencies (<10 Hz) induce LTD. For associative stimulation, the resulting effect depends on the temporal order of pre- and post-synaptic action potentials  is among the best understood molecular mechanisms underlying neuronal plasticity. It is characterized by a long-term increase of synaptic transmission following repetitive stimulation. LTP can be induced by tetanic stimulation at high frequencies, or by associative pre- and postsynaptic stimulation  by repeatedly presenting the identical visual or auditory stimulus and compared the brain's response before and after stimulation using electroencephalography (EEG)  from previous studies on response potentiation in the visual cortex as a starting point  participated in this study. Subjects had no history of psychiatric or neurological diseases and were not under medication at the time of the experiment. To avoid daytime specific effects, all experiments were performed in the afternoon. Written informed consent was obtained from all subjects prior to the experiments. The study protocol was approved by the ethics commission of the University Medical Center Freiburg , in agreement with the Declaration of Helsinki.As mentioned in the introduction, we adapted existing paradigms. In contrast to Clapp et al. , checkerFor the probe sessions, the design resulting from optimizing efficiency nearly matched a typical block design . Additionally, eight letters were presented as catch trials during the HFS period. Before the post-session, a pause of 2 min without visual stimulation was included to account for potential visual aftereffects . For fMRI, T2*-weighted gradient-echo echo-planar images (EPI) covering 192 × 192 × 81 mm were collected with the following parameters: TE: 32 ms, TR: 1920 ms, flip angle: 75°, matrix size: 80 × 80 × 28, voxel-size: 2.4 × 2.4 × 2.88 mm, gap: 20%, descending sequence, individually tilted to comprise the occipital cortex and the thalamus. Images were acquired continuously through probe and HFS periods. During reconstruction, scans were corrected for motion and distortion artifacts, based on a reference measurement  equipped with a 12-channel head coil. A T1-weighted whole-brain image was acquired as high-resolution structural reference for normalization and to exclude abnormalities (http://www.fil.ion.ucl.ac.uk). The volumes were spatially realigned and normalized to the Montreal Neurological Institute (MNI) reference brain, using the normalization parameters estimated during segmentation and normalization of the coregistered T1 anatomical scan , this removes a linear component from the variance explained by the main regressor . We focused on decreases during the pre- and post-HFS sessions, as this was reported in a previous EEG study . Significant clusters were characterized by their peak-coordinate in MNI space and their cluster extent. To illustrate spatial extent and heterogeneity of individual effects of potentiation, we combined all binarized clusters within the mask of the visual cortex, significant at the individual subject level in one figure.At the multi-subject level, the respective individual parameter maps  were entered into one-sample p et al. . To thisx, y, z: , MNI-coordinates).To illustrate signal changes during the HFS period and the probe sessions, we plotted the signal change in primary visual cortex, defined by a sphere (radius: 8 mm) around the peak voxel obtained from the main effect of flashes across both sessions in all subjects (SD: ±94 ms) post-HFS: 655 ms (SD: ±119 ms)].The behavioral data analyses revealed that the subjects correctly responded to 98.2% (range: 90–100%) of the catch trials . There was no difference between the response time of the pre- and the post-HFS session [mean response time pre-HFS: 651 ms . Contrary to that, the inverse contrast revealed a significantly stronger response to the flashes in the pre-HFS compared to the post-HFS session  compared to the first (pre-HFS) session at the multi-subject level  checkerboard flashes in order to induce an increased neuronal response. Contrary to expectations, activations related to pattern flashes, and also to the visual presentation of letters and to button presses were p < 0.01 at the voxel level from Clapp et al.  is coupled with the EEG signal during checkerboard reversals  spre- and post-sessions at the group level.Concerning the interval between flashes during the probe phase, studies using pattern flashes found that frequencies of 1 Hz (Teyler et al., The duration and stimulus frequency of the HFS period remained identical to related studies (Clapp et al., As we were initially interested in the inter-individual heterogeneity of SRM, we performed a single-subject analysis. Although activations during the HFS period itself indicated stable responses in all subjects (see Supplementary Figure post-hoc exclusion of subjects without a response potentiation (Stefan et al., Taken together, our results indicate a high heterogeneity of LTP-like effects in terms of the location but also their direction. This mirrors studies using SRM paradigms in the auditory system that produced incongruent results (Clapp et al., However, a recent study applied PAS-TMS, Theta burst stimulation and anodal dDCS and found very few subjects homogeneously responding to all potentially LTP-inducing interventions (López-Alonso et al., in-vivo.A better understanding of factors leading to a high variability of LTP-like effects, not observed in LTP at the cellular level, is essential to compare different LTP-like effects, to identify the underlying mechanisms and to establish tools to quantify individual synaptic plasticity The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The incidence of pancreatic cancer has been increasing while its 5-year survival rate has not changed in decades. In the era of personalized medicine, immunotherapy has emerged as a promising treatment modality in a variety of malignancies, including pancreatic cancer. This review will discuss the unique pancreatic tumor microenvironment, including the cells and receptors that transform the pancreas from its normal architecture into a complex mix of suppressor immune cells and dense extracellular matrix that allows for the unrestricted growth of cancer cells. Next, we will highlight the recently completed immunotherapy clinical trials in pancreatic cancer. Finally, we will explore the on-going immunotherapy clinical trials and future directions of this engaging and changing field. Despite intensive research efforts to better understand its tumor microenvironment, the prognosis of pancreatic cancer remains dismal , 2. The Immunotherapy is one of these novel strategies that has been under investigation in a variety of cancers. This review will focus on pancreatic cancer from an immune perspective, describing its immune microenvironment and the completed and ongoing clinical trials in this area.RAS oncogene drives an inflammatory program that establishes immune privilege in the pancreatic tumor microenvironment (PTME) [Pancreatic cancer is unique from an immunological perspective. First, intratumoral effector T-cells are rare, in contrast to many other solid tumors for which infiltration of effector T-cells is often prominent , 5. Secot (PTME) . Third, t (PTME) , 5. Fourt (PTME) . This det (PTME) , 8, 9. St (PTME) , 8, 9. Tt (PTME) . Accordit (PTME) .It is useful to examine the immune cells and receptors in pancreatic cancer based on their role in the development of an immune response and their correlation with prognosis. There are two immunological processes that determine the immune response against cancer cells: the effector process and the suppressor process. These cell types are summarized in Table An increased number of NK cells have been shown to be associated with a better prognosis in a small set of 13 patients with pancreatic cancer , presumaThe presence of TILs in pancreatic cancer has been well described and it may represent the most important element in PTME , 14, 5. The presence of DCs in the PTIM is essential in order to generate an anti-tumor immune response and, similar to TILs, is associated with a better prognosis in pancreatic cancer , 17, 18.Is a co-stimulatory molecule that is expressed on T-cells and binds to its ligand (CD40L) on APCs, leading to the activation of lymphocytes . In a reIs a member of the TNF receptor family found on T- cells and acts as a secondary co-stimulatory molecule as it requires other co-stimulatory molecules to be expressed first prior to its activation. The expression of OX-40 is found in high concentrations on activated T-cells. In pancreatic cancer increased levels of OX-40 was reported to correlate with better survival but this data needs to be validated on a larger scale .Is also a member of the TNF receptor family and found on T-cells and NK cells. Upon interaction with its ligand 4-1BBL, it promotes T cell activation, particularly CD8 lymphocyte. 4-1BB also acts as a secondary co-stimulatory molecule, similar to OX-40 , 21. HowThese cells express the program cell death ligand (PD-L1) which is involved in immune suppression and T-cell apoptosis as described below. In PTIM, the presence of TAMs and it is association with poor outcomes and more frequent metastases has been well described , 13.T-regs are other subsets of TILs known for their immunosuppressive activity through the release of cytokines including TGF-β and IL-10. Based on few retrospective analyses, pancreatic tumors with low numbers of T-regs were found to have a significantly better survival compared to tumors with high numbers , 22, 23.In pancreatic cancer, these cells were found in pre-malignant growths and increased in concentration as cancer cells grow suggesting a direct role in immune suppression and unrestricted cancer cell growth , 16, 24.Fibroblasts respond to a variety of molecules including CXCL12 (C-X-C motif chemokine 12) and produce VEGF  to stimulate angiogenesis in response to hypoxia or inflammation , 25. In The role of mast cells in cancer has not been well-defined , 28, 29.Is an inhibitory receptor that belongs to the B7-receptor family and interacts with its ligand PD-L1 (B7-H1) to down regulate signals by T-cells, leading to the induction of apoptosis in activated T-cells . PD-1 isIs a co-inhibitory molecule found on T- lymphocytes that deactivates these cells to induce apoptosis in response to interaction with APCs. This negative feedback loop is a key to normal immune function to prevent over-stimulation of T-cells and damage to healthy cells but is stimulated inappropriately in pancreatic cancer to create a microenvironment that promotes cancer growth rather than its recognition. CTLA-4 is overexpressed in pancreatic cancer cells and its overexpression was found to correlate negatively with survival in patients who underwent surgical resection .Is a chemokine that is found in high concentration in pancreatic cancer and is involved in fibroblast migration and proliferation. The increased concentration of CXCL12 in the cancer microenvironment creates a network of dense stroma restricting immune cells migration and recognition of cancer antigens. Feig et al., showed that the inhibition of this chemokine resulted in increased T-cell infiltration into pancreatic cancer in murine models .Is a transmembrane protein that is involved in the regulation of Th1 lymphocytes. The interaction between TIM-3 and CD4 Th1 cells has been studied in pancreatic cancer patients and found to correlate with tumor vascular invasion . HoweverIs an important T-cell regulator that interacts with MHC class II molecules expressed on APCs. This interaction promotes activation and maturation of DCs but negatively regulates effector T-cells. LAG-3 has been shown to be necessary for T-regulatory cell activity and represents a novel target for therapy in pancreatic cancer .Is an enzyme involved in cleaving tryptophan into kynurenine. Tryptophan is required for T-cell activation and kynurenine leads to T-regs differentiation and chemotaxis . IncreasAre immune modulating glycoproteins that are overexpressed in pancreatic cancer cells and thought to be involved in T-cell homeostasis. These glycoproteins have been shown to promote immune suppression in pancreatic cancer by promoting Th2 and T-reg transformation, restricting DC maturation and stimulating stellate cells , 36. TheIs a member of the B7 ligand family that can be induced in activated dendritic cells, monocytes, and T cells leading to decrease Th1 type response and cytokine production. B7-H3 expression was found to correlate with lymph node metastases and advanced pathologic stage in patients with pancreatic cancer . The inhCancer vaccines aim to stimulate the immune system against tumor cells by generating humoral and/or cellular immune responses. Many forms of cancer vaccines exist but generally they can be divided into synthetic and cellular-based vaccines.Synthetic vaccines are typically made from whole protein or peptides that match a pre-determined antigen to induce a T- cell response. Despite multiple large trials targeting MUC1 –43, VEGFCellular-based vaccines use cancer cells (either whole cells or cell lysates) as the source of the antigens, allowing the immune system to utilize multiple antigens rather than a single epitope. Overall the results of these trials are encouraging; however, each included a small number of patients making it difficult for meaningful interpretation. Cellular based vaccines can be divided into autologous or allogeneic vaccines based on the source of the cells.In this process, the patient’s own dendritic cells are isolated and pulsed with a specific antigen before being re-infused back to the patient. Multiple studies have used this vaccination approach targeting variety of antigens. MUC1 is one of the most targeted antigens given its overexpression in pancreatic cancer and its association with tumor invasion and metastasis. The clinical outcome of this vaccination method was not impressive; however, an interesting correlation with immune biomarkers was identified such as increased CD38 (a marker for activated lymphocytes)  and decrp = 0.01). Interestingly, patients who derived the most benefit of this combination were patients who received over 3 doses of the vaccine  and patients who received at least 2 previous chemotherapy regimens  [In this method of vaccination, a pancreatic cancer cell line is stimulated, usually with GM-CSF, in order to elicit an immune response when administered to patients with pancreatic cancer. A group at John Hopkins used this form of cancer vaccine (GVAX) in combination with 5-FU or chemoradiation in the adjuvant setting and showed an increase in PFS and OS in patients who developed an immune response against mesothelin , 61. The= 0.001) . This coex vivo then re-infused back to the patient. Based on the source and the method used for T-cell activation, adoptive T- cell transfer could be classified into: Tumor infiltrating lymphocytes (TILs), engineered T- cells that express a specific cancer T-cell receptor (TCR), and T- cells that express a chimeric antigen receptor (CAR). This methodology of immunotherapy had gained a lot of attention recently due to promising clinical outcomes in hematological malignancies. However, the efficacy of adoptive T- cell transfer remains to be determined in solid tumors including pancreatic cancer. Kawaoka et al., investigated MUC1-specific cytotoxic T lymphocytes (CTLs) in 28 patients and showed 19 % 3-year survival in patients with resectable disease along with increased effector lymphocytes and decreased T-regs [In this approach the patient’s T-cells are expanded and activated d T-regs . More red T-regs . The carImmune checkpoint inhibitors represent a paradigm shift in cancer treatment due to their promising clinical activities in melanoma and other malignancies , 67. HowDue to the lack of meaningful clinical benefits of cancer vaccines, the potential positive immunological effect of chemotherapy and radiation therapy, and the promising outcomes of immune checkpoint inhibitors, the focus has shifted towards combining these modalities. Gemcitabine, a standard chemotherapy that is used traditionally to treat pancreatic cancer, has been found to mediate immunological effects such as tumor associated antigen cross presentation by dendritic cells and the induction and expansion of cytotoxic T cells responses in addition to reduce the number of myeloid suppressor cells , 73. RadDespite the ongoing efforts outlined in this review, the prognosis of pancreatic cancer remains dismal. With the recent progress in cancer immunotherapy, there are glimmers of hope in new immune targets with more being identified each year. These advancements are moving from the bench to the bedside at a rapid pace, with the hope of translating into improvements in clinical outcomes. We believe that immunotherapy represents a promising modality in pancreatic cancer. However, there still remains much to be learned about the pancreatic immune microenvironment and its role in the immune escape of cancer cells. In order to develop an active strategy to enhance the immune response against pancreatic cancer that could be translated to a promising clinical outcome we must focus our efforts on increasing the density of the intratumoral effector T-cells; decreasing or inhibiting the immunosuppressive cells and receptors; and understanding the role of the stromal reaction and its interaction with pancreatic cancer immune microenvironment. Indeed, the recent encouraging data of patients with mismatch-repair deficient colorectal cancer responding to pembrolizumab requires further investigation, particularly as it may be relevant for a small number of patients with pancreatic cancer . Improve


Drosophila melanogaster, Caenorhabditis elegans, and Danio rerio. Moreover, a recent study examining the immune system of planarian will be discussed.The strategies evolved by pathogens to infect hosts and the mechanisms used by the host to eliminate intruders are highly complex. Because several biological pathways and processes are conserved across model organisms, these organisms have been used for many years to elucidate and understand the mechanisms of the host-pathogen relationship and particularly to unravel the molecular processes enacted by the host to kill pathogens. The emergence of RNA interference (RNAi) and the ability to apply it toward studies in model organisms have allowed a breakthrough in the elucidation of host-pathogen interactions. The aim of this mini-review is to highlight and describe recent breakthroughs in the field of host-pathogen interactions using RNAi screens of model organisms. We will focus specifically on the model organisms  Caenorhabditis elegans by Fire and Mello in 1998, the scientific community soon realized the potential of this tool for research  in their discoveries. During the Muller screen, 73% of the genes identified are positive regulators of the pathway (regulators that enhance the Drosophila JAK/STAT activity), whereas 75% of the genes identified during the Baeg screen are putative negative regulators (regulators that negatively regulate the Drosophila JAK/STAT pathway). Negative regulators were also discovered during in vivo genome-wide RNAi screens  pathway is known to play an important role in the control of a wide variety of biological processes. To investigate its mechanisms in the context of infection, two groups , transcriptional factors (DEAF1) involved in immune response, components required for phagocytosis of pathogens, components of IMD and NF-κB pathway, molecules required to activate Toll signaling pathway  and also the modulators of JAK/STAT pathway.Phagocytosis is a specific form of endocytosis involving the vesicular internalization of solids, such as bacteria. Utilizing this phagocytic capability of C. elegans exhibits a strong host defense response when challenged by different microorganisms  encoding proteins predicted to act in surface glycosylation, thereby influencing disease susceptibility  ligand. Using RNAi, they showed that nearly every gene in this network modulates the response to LPS in mouse macrophage cell lines.ms Table . For theC. elegans, only one Toll homolog (TOL-1) has been identified until now. In 2008, Tenor and Aballay  and a heat-shock protein (HSP-16.41) that is a member of the HSP family proteins needed for C. elegans immunity. Thus, for the first time, TOL-1 has been shown to play a direct role in the defense of C. elegans against pathogens.In C. elegans during S. enterica infection , which is encoded by the pmk-1 gene, is a prerequisite for activation of the Salmonella-induced programmed cell death (PCD). Inactivation of pmk-1 using RNAi completely attenuated Salmonella-elicited PCD. The same group confirmed the importance of the P38 MAP kinase pathway in the C. elegans immune response  pathway.” DAF-2 activity shortens life span through its inhibition of DAF-16, a forkhead transcription factor. Using microarrays, Kenyon's group  protein family. In the intestine of C. elegans was found to produce reactive oxygen species (ROS) as a powerful defense against infection. Through a combination of studies employing RNA interference and mutants to examine this ROS production . In zebrafish, four PGRPs have been identified, three of which have been cloned and named pglyrp-2 pglyrp-5, (pgrp-sc), and pglyrp-6  is not only linked to the immune response but also to apoptosis and developmental processes   that are present on the endosome  and Ifn-γ 2 (ifng1-2). Morpholino knockdown studies have shown a partially redundant function of ifng1-1 and ifng1-2 in mediating resistance of the zebrafish embryo to Escherichia coli and Yersinia ruckeri infections   knockdown disturbs ROS production, which increases the bacterial burden during Pseudomonas aeruginosa infection of zebrafish embryos  IFNs: Ifn-γ1  in restricting bacterial intracellular growth through non-canonical phagocytosis. This work established planarians as a suitable model organism for identifying anti-bacterial immune factors. Out of 18 antibacterial resistance genes identified they found the human orthologs for half of the genes including MORN2. In planarians, MORN2 protected against all strains of bacteria tested. Combining genetic and functional screen in planarians, researchers ascribed the function of human MORN2 to phagocytosis-mediated restriction of S. aureus, L. pneumophila and M. tuberculosis growth in macrophages. Further analyses supported the importance of MORN2 expression in promoting LC3-associated phagocytosis of M. tuberculosis and the targeting of bacteria to cathepsin-D-positive phagolysosomes. Complementary to these findings, MORN2 co-immunoprecipitated with LC3 and promoted the lipidation of LC3-I in cells challenged by autophagy-inducing stressors: LPS, IFN-γ and starvation.Quite recently, Abnave et al.  examinedGenetic screening in model organisms have proved to be a powerful and valuable tool in researcher's hand, and several remarkable studies on model organism (Tobin et al., Study of the immune response process has greatly benefited from the application of RNAi screens, as this approach is most successful when it incorporates a focused search for predicted pathway-regulatory proteins; moreover, the ability to adapt the RNAi method to several species, broadening the spectrum of meaningful data, has allowed researchers to deepen knowledge of the immune system in a way that was never possible before.The Guest Associate Editor, Philippe Soubeyran, declares that, despite having collaborated with author, Eric Ghigo, the review process was handled objectively. The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Ex-vivo resection and auto-transplantation avoids excessive bleeding and prevents ischemic related damage to the small intestine and other organs. Background: Tumors involving the root of the mesentery are generally regarded as “unresectable” with conventional surgical techniques. Resection with conventional surgery may end in life-threatening complications in these patients. ex-vivo resection of the tumors with involvement of small bowel mesentery followed by small bowel auto-transplantation.Objective: To share our experience of ex-vivo resection and auto-transplantation at our center were retrospectively analyzed. Methods: In this study, medical records of all the patients who underwent Results: The most common indication for the procedure in our series was locally advanced pancreatic carcinoma. Our survival rate was 50% with a mean±SD follow-up of 10.1±9.8 (range: 0–26) months. Causes of early in-hospital mortality were multi-organ failure, sepsis, and cerebrovascular accident. Recurrence of disease was noted in one patient while one patient developed hepatic metastasis after 20 months of surgery. Ex-vivo resection of the tumor and auto-transplantation is the surgical treatment of choice for the locally advanced abdominal tumors involving the root of the mesentery. Conclusion:  Before this, tumors involving the root of the mesentery were supposed to be “unresectable” by conventional surgical techniques without life-threatening complications [Ex-vivo resection of tumor and auto-transplantation not only prevent the development of short bowel syndrome but also provide better chance of obtaining tumor-free resection margin. The R0 resection is an independent factor for the long-term survival of the patient after surgery [Ex-vivo resection and auto-transplantation has long been in use for other organs such as heart, kidney and liver [The progress in the field of solid organ and multivisceral transplantation opens a window for resection of abdominal tumors involving the root of the mesentery by ications ,2. Conveications . Ex-vivo surgery ,5. Ex-vind liver ,7. ex-vivo resection of the tumors with involvement of small-bowel mesentery followed by small-bowel auto-transplantation.Therefore, in this study, we aimed to share our experience of ex-vivo excision of tumor and auto-transplantation at our center. Data of the patients were retrospectively analyzed from their medical records. All patients were thoroughly examined. Distant metastases were excluded by pre-operative radiological examination and during laparotomy in all but one patient in whom at the end of the procedure a single tiny metastatic lesion was discovered in the left lobe of the liver. Left lobe hepatectomy was done at the same operation to get a disease-free status. In another patient, the mass was found to be completely adhered to the left kidney so the left kidney was sacrificed to obtain tumor free resection margin. Two patients were referred from oncology clinic after receiving gemcitabine-based neo-adjuvant chemotherapy for surgical management of localized advanced carcinoma, but other patients did not receive this treatment. Patients who were discovered to have malignancy or positive lymph nodes on histopathology reports were referred to the Oncology Department for gemcitabine-based adjuvant chemotherapy.Between November 2010 and January 2013, 12 patients underwent Technical ConsiderationsComplete pancreatoduodenectomy with resection of the celiac and superior mesenteric artery was done in nine patients; in two patients with smaller tumors Whipple procedure was performed . In one The small-bowel resection, right extended hemicolectomy, splenectomy and retroperitoneal lymph node dissection (RPLND) were also performed in all patients; partial or complete gastrectomy was done in 11 patients. All tissue specimens were sent for histopathological examination.In our series, seven patients required interposition vascular grafts for the reconstruction of hepatic artery or portal vein. Allograft iliac vessels from deceased donors were used in the first two patients while in five other patients, autologus saphenous or external jugular veins were used.ex-vivo resection; liver and small-bowel were then auto-transplanted after tumor removal by bench surgery. This technique had an increased risk for developing complications of anhepatic phase, so we changed the surgical strategy. In other patients, the arterial inflow of the liver was replaced with a temporary interposition graft from infrarenal aorta to the proper hepatic artery using autologus saphenous vein before resection of the organs. Later, the proximal end of this bypass graft was repositioned and anastomosed to the stump of the resected celiac artery before small-bowel auto-transplantation. There were various techniques for approaching the common hepatic artery involvement. In the first two cases, the liver was also resected during ex-vivo resection of the tumor and auto-transplantation, males outnumbered to females in a ratio of 2:1. The mean±SD age of patients at the time of surgery was 45.7±14.6 (range: 17–73) years. Out of 12 patients who underwent Ten patients had pre-operative diagnosis of locally advanced pancreatic carcinoma, one had gastrointestinal stromal tumor (GIST), and another had retroperitoneal rhabdomyosarcoma . Two patOut of 12 patients, 10 had small bowel auto-transplantation; the remaining two had combined liver and small bowel auto-transplantation. After reperfusion, both liver and small bowel reimplanted grafts regain normal color and perfusion.The mean±SD total procedure time was 714±162 (range: 540–960) minutes. Total cold ischemia time varied between 60 and 210 (mean±SD: 160±68) min; the total warm ischemia time varied from 25 to 65 (mean±SD: 39±18) min. The mean hospital stay was 9.7 (range: 1–24) days. The two patients who received deceased donor vascular grafts were on immunosuppressive drugs. Both developed surgical complications and died of complications—one developed severe sepsis, probably due to over-immunosuppression; the other developed vascular graft thrombosis and small-bowel gangrene. After this event, we stopped using allograft interposition grafts.The survival rate was 50%. Six of our patients died during a mean±SD follow-up of 10.1±9.8 (range: 0–26) months. Three patients died during the initial hospital stay. Causes of in-hospital deaths were multi-organ failure, severe sepsis and cerebrovascular accident. Three patients died two months, three months, and six months after the initial surgery. Causes of late mortality were thrombosis of the allograft, late portal vein thrombosis and recurrence of disease, respectively. Nine patients with malignant disease survived more than six weeks. All of them were referred to oncologist for chemotherapy. They all received gemcitabine-based chemotherapy. One patient developed recurrence of disease within six months and died as mentioned above; one patient of pancreatic carcinoma developed multiple hepatic metastasis 20 months post-surgery. This patient had been referred to the oncologist for the systemic chemotherapy. Tumors involving the root of the mesentery are almost always supposed to be “unresectable” by the conventional techniques of surgery for several reasons. In an attempt to get tumor-free resection margins in these patients, either total or subtotal enterectomies have to be performed ending in short gut syndrome. TPN therapy is an option but chronic TPN therapy predisposes these patients to develop life-threatening complications such as line-related sepsis and liver failure ,9. ex-vivo resection and auto-transplantation in which the University of Wisconsin solution is used to prolong the cold ischemia time without any detrimental effect on the autologous grafts. These tumors are in close proximity to the major abdominal vessels supplying solid organs and hollow viscera. During resection of these retroperitoneal tumors, mesenteric vessels may need to be tied or clamped at least temporarily to control bleeding in the operating field. Of note, intestinal epithelium is very sensitive to warm ischemia and ischemic changes may appear just after an hour of clamping of the mesenteric vessels during warm ischemia . On the ex-vivo tumor resection and auto-transplantation. Dissection can be done in bloodless field in a timely manner, and thus with better chances of obtaining tumor-free resection margins. Also reconstruction of vascular pedicles can be done if needed and large enterectomies can be avoided with less risk of developing short gut syndrome. After exenteration of abdominal organs, abdominal cavity and retroperitoneal space become clearly visible. Any residual tumor or suspicious area can then easily be resected out. Furthermore, hemostasis can be achieved more effectively.There are several advantages for ex-vivo resection of tumors involving the root of the mesentery and auto-transplantation [To date, only few studies have been published in the medical literature on antation ,2,12-16.The underlying diseases for which this procedure has been done in these studies were mesenteric fibroma, vascular dysplasia of the root of the mesentery, desmoid tumor, mesenteric carcinoid tumor, mesenteric cavernous hemangioma and some cases of pancreatic carcinoma. In our series, the most common indication for the operation was pancreatic cancer involving the root of the mesentery. Obviously, malignant tumors need lymph node dissection and a more extended resection compared to benign lesions, which may increase the morbidity and mortality rates in patients with impaired immune system due to malignancy. This may explain the relative high rate of mortality in our patients. Use of cadaveric grafts to replace arteries or veins made us to use immunosuppressive drugs with all their side-effects. The two patients in whom allogenic vascular grafts were used, developed complications and died. So, we have not used this kind of vascular grafts since then.In conclusion, with refinement in surgical techniques as well as using neoadjuvant and adjuvant treatments, this procedure may lead to further improvement in the prognosis of malignant tumors involving the root of the mesentery. Furthermore, this approach would become the standard surgical treatment for otherwise unresectable benign lesions of the small bowel mesentery.