Drosophila melanogaster and Drosophilasimulans. An important feature of the analysis is classification ofTFBS mutations according to the direction of their predicted effect on bindingaffinity, which allows gains and losses to be evaluated independently along thetwo phylogenetic lineages. The observed patterns of polymorphism and divergenceare not compatible with neutral evolution for either class of mutations.Instead, multiple lines of evidence are consistent with contributions ofpositive selection to TFBS gain and loss as well as purifying selection in itsmaintenance. In discussion, we propose a model to reconcile the finding ofselection driving TFBS turnover with constrained CRM function over longevolutionary time.Transcription factor binding site(s) (TFBS) gain and loss  is awell-documented feature of cis-regulatory module (CRM) evolution, yet littleattention has been paid to the evolutionary force(s) driving this turnoverprocess. The predominant view, motivated by its widespread occurrence,emphasizes the importance of compensatory mutation and genetic drift. Positiveselection, in contrast, although it has been invoked in specific instances ofadaptive gene expression evolution, has not been considered as a generalalternative to neutral compensatory evolution. In this study we evaluate the twohypotheses by analyzing patterns of single nucleotide polymorphism in the TFBSof well-characterized CRM in two closely related Drosophila species, Drosophila species. We found the patterns ofdivergence and polymorphism for two types of mutations—those inferred toincrease or decrease the binding affinity respectively—are not compatiblewith a neutral hypothesis. Instead, multiple lines of evidence suggested thatpositive selection has contributed to gain as well as loss of TFBS in the twolineages, with purifying selection maintaining existing TFBS in the population.Spacer sequences also showed signatures of negative and positive selection. Weproposed a model of CRM evolution to reconcile the finding of frequent adaptivechanges with constraints on long-term evolution.Transcription factor binding sites (TFBS) turnover (i.e. lineage-specific gainand loss) is a well-documented phenomenon in eukaryote cis-regulatory modules(CRM). The wide spread of the phenomenon and the appearance of conservedexpression patterns for diverged orthologous CRM led to the standing view thatthe observed gain and loss of TFBS were functionally and selectively neutral. Tothe contrary, genome-wide population genetics analyses have unequivocallyidentified signatures of positive selection acting in noncoding regions ingeneral, and particularly in 5′ and 3′ untranscribed regions ofgenes. To specifically test the neutral versus selection hypotheses for the TFBSturnover process, we analyzed natural variation patterns within and between twoclosely related  Gene expression in eukaryotes is generally controlled by transcriptional enhancers,also called cis-regulatory modules (CRM), which are short regions in the genomeconsisting of a cluster of transcription factor binding sites (TFBS) spaced byintervening sequences (spacers). Individual TFBS have been shown repeatedly to berequired for CRM function, yet surprisingly they evolve rapidly and are frequentlygained and lost in evolution, attributes that have been demonstrated for a largenumber of CRM and transcription factors Drosophila CRM evolution (as exemplified byeve stripe 2 enhancer), whereas a model that assumes the doublemutant to be more fit than the wildtype does.The gain or loss of a TFBS is unlikely to be functionally irrelevant, as repeatedlyshown in TFBS knockout experiments This theoretical finding raises the prospects for positive selection being animportant driving force of TFBS gain and loss. Instances of directional selectionhave been documented in cases where a novel regulatory regime is favored Bicoid-dependent hunchback enhancer and theeven-skipped stripe 2 enhancer. This test can also be used todetect positive selection, although its power is limited due to the mixed signalwith purifying selection, which is expected to be dominant in most cases.Several different approaches have been designed to detect and quantify selection inthe system. One of them has been to consider the genome-wide ensemble of TFBS asevolving at mutation-selection balance, with the fitness of each instance of TFBSbeing strictly determined by its binding energy Drosophila species, D.melanogaster (mel) and D. simulans(sim). The short evolutionary distance between the two speciesensures unambiguous alignment for noncoding sequences and also allows one to capturethe potentially rapid dynamics of TFBS gain and loss. A notable challenge instudying TFBS turnover is assembling a high quality set of TFBS that are preciselydefined and contain few false positives. Large numbers of potential TFBS can beidentified by methods involving genome-wide scans, such as computational predictionor ChIP, but these approaches generally include a large fraction of false positives,thus reducing their attractiveness for investigating the mechanisms of binding siteturnover  both toidentify TFBS within footprints and to predict the magnitude of binding energydifferences among variant alleles. Our bioinformatic and experimental validationsshowed that the PWM used in this study provide reliable and unbiased estimates forthe direction of binding affinity change in both mel andsim  and divergence in 645experimentally identified TFBS for 30 transcription factors in 118 autosomal CRM, all ann andsim .mel andsim were polarized with outgroup sequences from D.sechellia, D. yakuba and D. erectausing PAML  but lost insim. A set of empirical cutoffs were determined for each TFbased on the range of PWM scores among its footprint sites, which producedsimilar results . This is not unexpected, given that allfootprint TFBS were identified as being present in mel and thedataset doesn't include sim-specific TFBS. We predictedthat identification of TFBS by computational methods would produce a more evenpattern of gains and losses in both lineages. We tested this prediction forthree TF  using a stringent cutoff procedure and for each TF we founda similar total number of predicted binding sites in the two lineages  test frameworkmel impacts theanalysis in two ways. First, gains of TFBS can be observed inmel but not losses, while the reverse is true insim. Therefore, even though similar processes are mostlikely operating in both species, our evolutionary analysis of binding site gainwill focus on changes in the mel lineage, whereas losses willbe restricted to changes in the sim lineage.The fact that all footprints were identified in mel. Thisarises because mutations in TFBS were sampled conditioned on the TFBS beingdetected in mel and affinity-changing mutations inmel, in turn, have the potential to affect thedetectability of the TFBS. Depending on whether the derived mutation isaffinity-increasing or affinity-decreasing, two distinct biases are introducedin the expected neutral frequency spectrum , where the loss of a TFBS is observable.To delineate the effect of ascertainment from that of selection for theaffinity-increasing and affinity-decreasing mutations, we compared the observed  values .For affent bias . This seFor affinity-increasing mutations no amount of ascertainment under our model canaccount for the observed relative excess of substitutions  red. We affinity . These rTo further investigate evolutionary forces acting on the segregating mutations inTFBS in the population, we utilized the site frequency spectrum, for which wegenerated the neutral expectations for affinity-increasing andaffinity-decreasing mutations separately under ascertainment, withutations . Howeverutations . Thus weutations . Thus, wsim are notinfluenced by the ascertainment because the identification of TFBS inmel is independent of the effect of mutations fixed orsegregating in sim. However, the inclusion of binding sitesgained in mel may confound the analysis as their orthologoussequences in sim may have evolved under less or different kindsof selective constraints. We thus restricted the analysis to footprint TFBSpredicted to be present in the mel-sim common ancestor, wherewe found a significant excess of substitutions for the affinity-decreasingmutations compared to the synonymous No-Change class  and thedifficulty in validating the biological functionality of individual binding sites.While many genome-wide datasets for TFBS are becoming available, several propertiesof the Drosophila, but not the biological orfunctional basis for this selection. In this study, we distinguished mutations inthe footprint sites by their functional impact – either increase or decreasethe binding affinity of the corresponding TF – and observed different patternsof polymorphism and divergence between the two classes. For example, we found thataffinity-decreasing mutations are on average more deleterious among new mutationsthan affinity-increasing ones, as revealed by a comparison of the ratio between thetwo classes in polymorphism with the expectation from mutational input. Suchdistinctions were not observed when mutations were grouped in other ways irrelevantto the function of TFBS . For these reasons we think the evidence supports ourspecific model of selection acting on binding site gain and loss as opposed to anunidentified functionality in noncoding sequences in general. The mechanism ofselection we described here for well-annotated TFBS could in principle be actingmore broadly across noncoding regions inasmuch as noncoding DNA is often associatedwith proteins binding.Genome-wide studies have identified signals of both positive and negative selectionin noncoding sequences in hunchback andbcd binding sites. To our knowledge, this is the first timethat accurate measurements have been made on population-level variants in TFBS. Wefound that PWM scores correctly predicted the measured direction of affinity changefor 21/25 mutations investigated. Of the four mutations that PWM predicted the wrongdirection, three have effect sizes predicted to be close to zero. The PWM-basedprocedure, therefore, may not be accurate for small predicted differences in bindingaffinity. Taking these results into consideration, we employed a binaryclassification of mutations with PWM differences exceeding a threshold requirementrather than using quantitative predictions of all PWM score differences as a basisfor our analysis.Our ability to correctly categorize mutations into affinity-increasing oraffinity-decreasing categories hinges on the accuracy of PWM predicted affinitydifferences. To investigate this issue, we employed a state-of-the-art microfluidicstechnique, MITOMI mel derived PWM to scoresim TFBS binding affinity. Transcription factor proteinevolution between the two species, if it occurred, could lead to underestimation ofbinding affinity in sim, although the effect should be similarlyapplied to both substitutions and polymorphism and thus is not expected to cause arelative excess of the former as observed in the sim data.Nevertheless, we show two lines of arguments that suggest this is not the case inour study: first, for the 30 TF whose binding sites we investigated, the DNAbindings domains and other functionally annotated domains are completely conservedexcept for one biochemically conservative amino acid difference (Asp/Glu) inDorsals RHD domain . To further support this argument, we derived PWM using MEME from themel footprint sites as well as their aligned sequences insim. As shown in mel PWM or thesim PWM, contrary to what would be expected if TF sequencespecificity had evolved between the two species. Therefore we consider it veryunlikely for the 30 TF included in this study to have undergone significantevolution in their sequence specificity. In addition, because the SELEX derived PWMproduce consistent results with the footprint derived ones . We can think of only three mechanisms by whichTFBS loss can occur at an appreciable rate: (1) there is loss of constraint; (2) apair of tightly linked compensatory mutations creates an effectively neutral allele;or 3) positive selection drives the loss of TFBS. Our second finding – asignificant excess of substitutions compared to the neutral class foraffinity-decreasing mutations in sim – is consistent onlywith positive selection for TFBS loss. Occasional adaptive loss of a TFBS is notinconsistent with more ubiquitous selection to maintain binding sites Our population genetics analysis identified three major forces in TFBS evolution.First, we found functional TFBS were selectively maintained in the population bypurifying selection, as revealed by a frequency spectrum skewed towards rarevariants for affinity-decreasing polymorphism in mel. Collectively, the three findings indicate that naturalselection is extensively involved in the maintenance, gain, and loss of TFBS. Thisconclusion challenges the prevailing view of a neutral TFBS turnover process Our third finding is positive selection contributing to the gain of TFBS, as revealedby a significant excess of substitutions for affinity-increasing mutations inDrosophilaeven-skipped stripe 2 enhancer, it demonstrated clear functionaldifferences between CRM that were previously believed to have the same spatialpattern of expression We think that a selectionist interpretation of the turnover process is plausible forseveral reasons. First, the assumption of CRM functional stasis, which is the mainargument for the neutral  view, is not well supportedexperimentally. Reporter transgene assays, in particular, are limited in theirquantitative resolution, and yet even in these studies, repeatable differences werefound between orthologous CRM Drosophila TFBS turnover rate, theparameterization of the model requires that the double mutant allele have higherfitness than the non-mutant allele, making it a directional selection model. Thissupercompensatory scenario could produce signatures of positive selection both forbinding site gain and loss, the latter occurring because the fixation of adeleterious mutation in an existing TFBS will have the appearance of beingpositively selected as it hitchhikes to fixation on the selectively favored allele.However, this scenario is biologically unrealistic, as it requires the secondmutation (the gain of a TFBS) to be positively selected only on the background ofthe first mutation.Second, compensatory neutral evolution cannot account for the patterns of variationobserved in this study. According to this model, affinity-decreasing mutationsshould in general be deleterious but occasionally become “effectively”neutral when a second compensatory mutation occurs in the CRM of the mutant allele.A mixture of deleterious and compensatory mutations, even if the latter is common,may bring patterns of polymorphism and divergence close to a neutral scenario, butcannot produce a signature of positive selection as observed for both classes ofmutations in our analysis. In addition, analytical modeling of the compensatoryevolution of TFBS finds that the waiting time for a turnover event is long ifcomplete neutrality of the compensating mutations is assumed Drosophilaspecies have been studied as potentially adaptive traits; if egg length evolves,genes such as eve whose expression pattern need to scale with theembryo may need to change its CRM to adapt to the new context Drosophila, with changes in the sign of selection coefficientoccurring at nearly the rate of neutral evolution As an alternative, consider the following model of positive selection on CRMstructure/function. We propose that for CRM with large numbers of interactingpartners, the network of cis- and trans-factors will inevitably be constantlyevolving – due to both direct selective pressures imposed on the CRM orindirect effects caused by adaptations in other components of the network. Forexample, egg length variations between and within Drosophila eve stripe2 enhancer D. pseudoobscura rescues a mel evestripe 2 enhancer deficiency almost as well as the native melenhancer and substantially better than ones from much more closely related species.As a result, a total of 645 TFBS for these 30 TF were included for analysis.REDfly D. melanogasterreference genome listed in D. melanogaster(mel), D. simulans (sim),D. sechellia (sec), and two outgroupspecies, D. yakuba (yak) and D.erecta (ere). D. sechellia is asister species to D. simulans and is included to compensate forthe low sequence completeness in the reference sim genome. Wethen used the baseml module in PAML 4.4c mel andsim determined by the most probable ancestral state  at each position, or summing over the posterior probabilities of all fourpossible states (full Bayesian approach). The two methods produced essentiallythe same results and therefore we only presented results using the most probableancestral state. A maximum parsimony method was also investigated and was foundto produce consistent results.For each of the 118 CRM (coordinates in dm3 of D. melanogaster lines (http://www.hgsc.bcm.tmc.edu/projects/dgrp/) and six D.simulans lines (http://www.dpgp.org/). We alsocompiled the genome sequences of 150 coding regions corresponding to the targetgenes of the CRM listed in REDfly, for the purpose of compiling synonymous andnonsynonymous changes. For these data, we used codeml module in PAML 4.4c toreconstruct the ancestral sequence states following otherwise the same procedureas described above for CRM regions.For polymorphism analysis, alignments for the same 118 CRM regions were obtainedof a population sample of 162 et al. identified a single best motif for the REDflyfootprint sites. For comparison, we also constructed five PWM  from SELEX  data(kindly provided by Mark Biggin). We ran MEME PWM for 30 TF  were obtained from Consider a mutation at the et al.mel and sim. Data were analyzed in GenePix6.0, R, and Prism 5.0. We found that the PWM we used correctly predicted thedirection of change in 21/25 cases  of mutations in the TFBS wereconsistently classified after excluding nine mutations with small predictedeffects by either PWM , using patser v3e . To determine whether a sequence is a binding site ornot, we established two sets of cutoffs for PWM scores. First, we used PWM scoreTo examine the extent of binding sites gain and loss between the two species, wecalculated PWM scores  results .mel-derived PWM might be over-optimized sothat they would favor mel over sim sequencesindependent of the binding affinity differences, we ran MEME on bothmel footprint sites for three TF  and theirsim orthologous sequences with the same parameters. The twoset of ÒorthologousÓ PWM were then applied to score the observedvariations in the TFBS of the three TF for comparison : Next-generation sequencingdata produce variable coverage. To estimate the site frequency spectrum, foreach variable site  with a coverage greater than orequal to 150 (maximum is 162) we randomly chose 150 and combined the counts foreach frequency class (from 1/150 to 149/150).Site frequency spectrum , (C) and (E), Proportions of predicted matchesto Hunchback (hb), Bicoid (bcd) or Krpple (Kr) PWM that aremel-specific, sim-specific or sharedin both species in each BCD or KR regulated enhancer region (defined asregions that contain at least one mel footprint site for the TF). Numbers inthe white bar indicate the number of shared predicted sites. (B), (D) and(F) are similar to (A),(C),(E) except that they include 200 bp flankingsequences on each side of an enhancer.(PDF)Click here for additional data file.Figure S2hb PWM compared toin-vitro direct measurement by MITOMI. MITOMI experiments were performed asdescribed in the methods. Each mutation was measured in two oligonucleotidescarrying the original and mutant nucleotide respectively. The two dashedlines indicate the cutoff we applied in the study.Binding affinity change predicted by (PNG)Click here for additional data file.Figure S3PWM based on mel footprints and SELEX PWM produce consistent results. Eachpoint represents one substitution and its x, y values are the estimates ofits effect on binding affinity using the footprint PWM or the SELEX PWM,respectively. 33/34 strong-effect substitutions are consistently assigned bythe two sets of PWM into either affinity-increasing or affinity-decreasingcategories.(PDF)Click here for additional data file.Figure S4mel. Each box represents a TFBS, where orange indicatesrelatively strong binding affinity while greens indicates weak affinity.Each column is an alignment of a sample of six mel alleleswith the inferred ancestral allele. In the first column, a fixedaffinity-decreasing mutation in mel with a relatively largeeffect makes the TFBS not detectable as a footprint. In column 2 and 3, theaffinity-decreasing mutations are not fixed but segregating, therefore theprobability of not detecting the TFBS is proportional to the derived allelefrequency . Column 3–6 illustrate the situation foraffinity-increasing mutations, where the substitutions are always detectablebut the segregating mutations are detected with higher probability when thederived allele frequency is low. The last two columns represent cases whereboth alleles are detectable. To incorporate the uncertainty in thedetectability of the low-affinity allele, we define a parameterf for the probability that the weak allele is notdetectable.Impact of ascertainment on the detectability of a mutation in(PDF)Click here for additional data file.Figure S5Site frequency spectra for different classes compared to the neutralexpectation (A) Non-synonymous; (B) Synonymous No-Change (C)Preferred-to-Unpreferred; (D) Unpreferred-to-Preferred; (E) Spacers in CRM.Black: neutral expectation; Gray: observed site frequency spectrum.(PDF)Click here for additional data file.Figure S6mel. The proportion of lowfrequency class(es) for affinity-decreasing mutations compared to thetheoretical neutral expectation, the observed synonymous sites, or theexpected proportion for synonymous sites under ascertainment assumingRelative excess of rare variants suggests purifying selection on affinitydecreasing mutations in (PDF)Click here for additional data file.Figure S7mel footprints (PWMmel) or their alignedsequences in sim (PWMsim) produce consistent results underour classification method. On the scatter plot each point represents asingle nucleotide mutation with its x, y values being the estimates of itseffect on binding affinity using either the mel PWM or thesim PWM, respectively. Green and red triangles aremutations occurring on mel or sim lineages. From thefigure, the PWM have very little biases with respect to scoring mutationsfrom the species where it is derived or the other species.PWM derived from (PDF)Click here for additional data file.Figure S8D.sechellia as an outgroup (bold and blue). The referencesequencing strain  is obviouslyinter-mingled with the other 162 lines. A similar procedure on 3 different10 kb sequences sampled from the genome produced similar shaped trees withthe lab line embedded among the 162 lines, although the exact orders ofbranches are not the same, reflecting different geneaologies between regionsin the genome.A geneaology tree based on 10 kb CRM sequences for 162 lines from DGRP andthe Berkeley reference sequencing strain. The tree is built in MEGA usingmaximum likelihood method, based on 10 kb sequence alignments. It is rootedwith one sequence from a closely related species (PDF)Click here for additional data file.Table S1CRM studied in this study.(PDF)Click here for additional data file.Table S2Percentage of gain and loss of TFBS predicted by two set of cutoffs.(PDF)Click here for additional data file.Table S3The pattern of excess for aff-dec mutations in sim is robust to choices ofcutoff.(PDF)Click here for additional data file.Table S4mel and sim.TF protein sequence divergence in different functional regions between(PDF)Click here for additional data file.Table S5PWM derived from SELEX sequences using MEME.(PDF)Click here for additional data file.Text S1Computational prediction of TFBS in CRM.(DOC)Click here for additional data file.Text S2Neutral expectations for the MK test and the site frequency spectrum underascertainment.(DOC)Click here for additional data file.Text S3Validity of transferring the footprint sites identified in lab strains to thenatural populations.(DOC)Click here for additional data file.


The fibromatoses are a group of benign fibroblastic proliferations that vary from benign to intermediate in biological behavior. This article will discuss imaging characteristics and patient demographics of the adult type superficial  and deep (musculoaponeurotic) fibromatoses. The imaging appearance of these lesions can be characteristic (particularly when using magnetic resonance imaging). Palmar fibromatosis demonstrates multiple nodular or band-like soft tissue masses arising from the proximal palmar aponeurosis and extending along the subcutaneous tissues of the finger in parallel to the flexor tendons. T1 and T2-weighted signal intensity can vary from low (higher collagen) to intermediate (higher cellularity), similar to the other fibromatoses. Plantar fibromatosis manifests as superficial lesions along the deep plantar aponeurosis, which typically blend with the adjacent plantar musculature. Linear tails of extension  along the aponeurosis are frequent. Extraabdominal and abdominal wall fibromatosis often appear as a heterogeneous lesion with low signal intensity bands on all pulse sequences and linear fascial extensions  with MR imaging. Mesenteric fibromatosis usually demonstrates a soft tissue density on CT with radiating strands projecting into the adjacent mesenteric fat. When imaging is combined with patient demographics, a diagnosis can frequently be obtained. The fibromatoses are a broad group of fibroblastic proliferations with a similar histologic appearance containing spindle-shaped myofibroblastic cells, dense deposits of intercellular collagen fibers, variable amounts of extracellular myxoid matrix, and compressed and elongated vessels . They va The superficial  fibromatoses arise from fascia or aponeuroses at palmar, plantar, penile (Peyronie disease), and knuckle pad locations. Of the superficial fibromatoses, palmar fibromatosis is the most common followed by plantar fibromatosis , 4. Palmar fibromatosis (Dupuytren disease) is the most common of the superficial fibromatosis, affecting 1%-2% of the general population and approximately 4% of the United States population –7. It wa The etiology of palmar fibromatosis is believed to be multifactorial, including components of trauma, microvascular injury, immunologic processes, and genetic factors. Patients are typically over 65 years of age and the process is rarely seen in children. Males are affected 3-4 times more often than females and the disease is more severe in men . Clinically, patients present with painless, subcutaneous nodules involving the palmar aspects of the fingers, usually the fourth and fifth digits , 11. The Surgical intervention remains the treatment of choice, typically a selective fasciotomy. The decision to undergo surgical excision is determined by both patient symptoms and the presence of flexion contracture greater than 20 degrees at the metacarpophalangeal (MCP) joint or greater than 30 degrees at the proximal interphalangeal (PIP) joint . A simpl Radiographs may be normal or demonstrate flexion (Dupuytren) contractures of the MCP and PIP joints . On ultrasonography, the subcutaneous nodules of palmar fibromatosis are generally hypoechoic, hypervascular, and superficial to the flexor tendons , 17. The Computed Tomography (CT) demonstrates isoattenuated to slightly hyperattenuated nodular areas of subcutaneous thickening. MR imaging (MR) demonstrates multiple nodular or band-like soft tissue masses arising from the proximal palmar aponeurosis to the flexor tendons . The nod Plantar fibromatosis (Ledderhose disease) (Morbus Ledderhose) occurs less frequently than the palmar lesion, with an incidence of 0.23% . In our  Although the lesion can occur in children , 22, inc Patients present with one or more subcutaneous nodules, which most frequently arise in the medial aspect of the plantar arch (78%) and can extend to the skin or deep structures of the foot. Nodules may be multiple in 33% of cases . Palmar  The treatment of plantar fibromatosis is often conservative and consists of footwear modifications aimed at relieving symptoms . Surgica Radiographs are frequently normal in patients with plantar fibromatosis. Lesion evaluation is most commonly performed with ultrasound and MRI. Sonographic imaging demonstrates a well-defined (64%) or poorly defined 36%) fusiform mass in the soft tissues adjacent to the plantar aponeurosis . Plantar6% fusifo CT images demonstrate a nonspecific soft tissue mass in the characteristic location with attenuation equal or higher than skeletal muscle . MR imaging may demonstrate well-defined or ill-defined superficial lesions along the deep plantar aponeurosis, which typically blend with the adjacent plantar musculature. With its superior soft tissue contrast, MR is the best modality to determine infiltration of the lesion into the surrounding tissues and therefore it is most helpful for preoperative planning. Lesions typically show heterogeneous signal (92%), which is isointense to hypointense to skeletal muscle on T1W (100%) and T2W (78%) sequences. If the lesion has increased cellularity and less collagen, the T2 signal is increased (22%) . The deg The deep fibromatoses are fibroblastic proliferations that arise within the deep soft tissues and are traditionally divided into extraabdominal, abdominal wall, and intraabdominal types. They demonstrate infiltrative growth and local recurrence but do not metastasize. Their biological behavior may be considered intermediate due to frequent local recurrence. Involvement of adjacent vital structures may lead to patient demise, particularly with neck and chest wall lesions. Recurrent extremity lesions may eventually require amputation for local control , 6. The  The overall incidence of desmoid type fibromatosis is two to four individuals per million each year , 32. Rel Synonyms for extraabdominal fibromatosis include musculoaponeurotic fibromatosis, extraabdominal desmoid, desmoid tumor, well-differentiated nonmetastasizing fibrosarcoma, and aggressive fibromatosis , 6. Extraabdominal fibromatosis is most common in patients between puberty and 40 years of age, with a peak incidence noted between the ages of 25 and 30 years. Less than 5% of patients are younger than 10 years of age , 35. While these lesions can occur almost anywhere in the body, they have a predilection for the upper torso including the upper arm (28%), chest wall/paraspinal 17%), and head/neck 10% to 23%). Other less common locations include the thigh (12%), knee (7%), buttock/hip 6%), and forearms (4%). Lesions in the head and neck often behave more aggressively and may surround the axillary vessels, trachea, and brachial plexus, limiting the extent of surgical resection [%. Other %, and fo7%, and h Although these tumors are usually solitary, synchronous multicentric lesions  are note Extraabdominal fibromatoses typically present as a slow growing, painless mass which can limit range of motion of a nearby joint and invade adjacent neurovascular structures. Decreased range of motion, neurologic symptoms, and pain are reported but are unusual at presentation. Lesions are typically between 5 and 10 cm in size . Extraab The treatment of extraabdominal desmoid is usually a wide-local excision as it has a high tendency to locally recur. Traditionally, positive surgical tumor margins upon resection have been reported to be associated with a higher local recurrence rate. However, more recent studies have suggested that positive versus negative microscopic margins do not make a difference in the overall local recurrence rate , 39. Adj The rate of local recurrence varies from 19% to 77% (average 40%) and is usually within two years of resection , 6.Radiographs usually appear normal, although there may be signs of a soft tissue mass such as the failure to visualize the radiolucent cleavage plane between the soft tissue and bone, structural changes of the adjacent bone such as scalloping, or rarely soft tissue calcification or ossification may be seen . Bone inScintigraphy with Technetium-99m pertechnetate (Tc-99 m) has been used as a tumor scanning agent for follow-up of extraabdominal fibromatosis. Focal tracer accumulation is noted on blood pool and delayed static images .Sonography of desmoid-type fibromatosis reveals a hypoechoic lesion –51. The On CT, extraabdominal fibromatosis appears as a nonspecific soft tissue mass. Unless outlined by fat, the margins of the lesion are poorly defined. The attenuation of these tumors is variable, and has been described as lower than, similar to, or higher than skeletal muscle. Lesions with a higher attenuation are noted to have significant collagen components. One author suggests that a high attenuation lesion which is hypoechoic on ultrasound is suggestive of a fibrous lesion . The lowMR is frequently the modality of choice to evaluate and stage extraabdominal fibromatosis. Extraabdominal desmoid is usually centered in an intermuscular location, often along the deep fascia; therefore, a thin rim of surrounding fat may be noted (split-fat sign). Lesions may be well defined (49% to 54%) or have irregular infiltrative margins 46% to 51%) . T1-weig% to 51% Three histopathological stages of desmoid tumor have been described. In the first stage, lesions are more cellular , with laThe morphology of these low-signal areas as prominent band-like regions Figures  is more Signal intensity on long TR sequences may have an implication on tumor recurrence, with a higher recurrence rate in lesions with high T2 signal . LesionsIn lesions undergoing radiation or drug therapy, MR surveillance has been used to assess response to treatment  with a pAlthough not routinely performed in most institutions, it has been reported that diffusion-weighted imaging may help differentiate desmoid tumors from malignant soft tissue tumors, with fibromatosis demonstrating a higher mean apparent diffusion coefficient (ADC) than malignant soft tissue tumors .Areas of low T2-weighted signal are not specific to fibromatosis and may be seen with other lesions. A differential diagnosis for soft tissue lesions with prominent areas of low-signal intensity on T1- and T2-weighted sequences includes desmoid type fibromatosis, densely calcified mass, pigmented villonodular synovitis (PVNS)/giant cell tumor of tendon sheath (GCTTS), elastofibroma, granular cell tumor, desmoplastic fibroblastoma, and malignant fibrous histiocytoma (MFH)/fibrosarcoma .FDG PET has been utilized to evaluated decreased metabolic activity of all types of deep fibromatosis during chemotherapy , 66. Des Abdominal wall fibromatosis is indistinguishable both grossly and histologically from extraabdominal fibromatosis and the relative frequency is similar. It is discussed separately because of its characteristic location and the tendency to occur in women of childbearing age  during or more frequently within the first year following a pregnancy and in women who use oral contraceptives . It is t Clinical presentation is typically palpation of a firm, slowly growing painless soft tissue mass. Endocrine factors are most highly implicated in this form of deep fibromatosis by the frequent occurrence of these tumors during or in the year following pregnancy. Estrogen receptors have been reported in 79% of these lesions  and they These lesions frequently recur locally. The rate of local recurrence is reported to be 15% to 30%, less frequent than extraabdominal desmoid (35% to 65%) , 77. SimThe radiologic features of abdominal wall desmoid, with all imaging modalities, are essentially identical to those of desmoid type fibromatosis in other locations. Involvement of the rectus abdominis muscle is most common Figures  and 10.  Intraabdominal fibromatosis  is a rare group of closely related deep fibromatoses that occur in the pelvis, mesentery, and retroperitoneum . Despite Pelvic fibromatosis occurs in the iliac fossa and lower pelvis. It presents as a slowly growing palpable mass asymptomatic or causing slight pain. It is often mistaken for an ovarian neoplasm. The lesion occurs most frequently in women 20 to 35 years of age . The small bowel mesentery Figures  is the m Pelvic fibromatosis shows a female predilection, whereas mesenteric desmoids demonstrate a slight male predilection (55%) . The age Intraabdominal fibromatosis demonstrates sporadic occurrence, but the incidence is increased in patients with familial adenomatous polyposis (FAP), trauma or hyperestrogenic states , 52, 83. The presenting clinical signs and symptoms of mesenteric fibromatosis are often related to the small bowel. Patients may complain of abdominal pain or a palpable abdominal mass or come to clinical attention because of complications such as gastrointestinal bleeding, small bowel obstruction, fistula formation, bowel perforation, or hydronephrosis , 85, 86. Treatment options include surgical excision, hormone therapy, nonsteroidal anti-inflammatory drugs, and cytotoxic chemotherapy, although the optimal treatment regimen is controversial . While sIntraabdominal fibromatosis may result in local mass effect with displacement of adjacent bowel loops on radiographs . SerosalMesenteric masses may be discovered incidentally when patients are being evaluated for complaints of abdominal pain or discomfort. The sonographic appearance of mesenteric fibromatosis is a solid, well-circumscribed mass of variable echotexture and homogeneity , 80, 83.Mesenteric fibromatosis usually demonstrates a soft tissue density with radiating strands projecting into the adjacent mesenteric fat Figures  and atteMost intraabdominal lesions are low or intermediate signal intensity on T1-weighted images and have heterogeneous low-, intermediate- or high-signal intensity on T2-weighted images. The relative amount of hyperintensity on T2-weighted images reflects the degree of high cellularity areas and myxoid stroma within the lesion. The intravenous contrast enhancement pattern of mesenteric fibromatosis with MR is variable. Lesions that do not significantly enhance with iodinated contrast material on CT scans have been shown to enhance with intravenous gadolinium on MR imaging , 86. In Mesenteric fibromatosis has been reported as a false positive during FDG PET for detection of metastatic disease , 92. Gardner syndrome is a variant of familial adenomatous polyposis (FAP) containing osteomas, thyroid cancer, epidermoid cysts, fibromas, sebaceous cysts, and desmoid tumors in addition to the colorectal adenomatous polyps . Gardner syndrome is usually diagnosed in adults 25 to 30 years of age  and is m Patients with Gardner syndrome may demonstrate intraabdominal, abdominal wall, or musculoaponeurotic fibromatosis . Of the  Although similar in imaging characteristics to the isolated lesions, intraabdominal fibromatosis and other desmoid tumors associated with Gardner syndrome tend to be smaller and multiple and occur in a younger patient population. The smaller size and younger patient population may be secondary to more frequent imaging and follow-up examinations performed in patients with Gardner syndrome. Complications secondary to desmoid tumor are the most common cause of death in patients with FAP who have undergone prophylactic colectomy (30.6%) . In summary, the deep and superficial fibromatoses vary from low- (higher collagen) to-intermediate (higher cellularity) T1 and T2-weighted signal intensity depending on lesion cellularity. Palmar fibromatosis may result in flexion (Dupuytren) contracture and plantar fibromatosis typically involves the medial cord of the plantar fascia and blends with the adjacent plantar musculature. A heterogeneous lesion (well-defined or ill-defined) with nonenhancing low-signal-intensity bands on all pulse sequences and linear fascial extensions  are highly suggestive of abdominal wall or extraabdominal desmoid . Mesente


To develop a customized short LOS (<6 days) prediction model for geriatric patients receiving cardiac surgery, using local data and a computational feature selection algorithm.Utilization of a machine learning algorithm in a prospectively collected STS database consisting of patients who received cardiac surgery between January 2002 and June 2011.Urban tertiary-care center.Geriatric patients aged 70 years or older at the time of cardiac surgery.None.Predefined morbidity and mortality events were collected from the STS database. 23 clinically relevant predictors were investigated for short LOS prediction with a genetic algorithm (GenAlg) in 1426 patients. Due to the absence of an STS model for their particular surgery type, STS risk scores were unavailable for 771 patients. STS prediction achieved an AUC of 0.629 while the GenAlg achieved AUCs of 0.573 (in those with STS scores) and 0.691 (in those without STS scores). Among the patients with STS scores, the GenAlg features significantly associated with shorter LOS were absence of congestive heart failure (CHF) , aortic valve procedure , and shorter cross clamp time . In those without STS prediction, short LOS was significantly correlated with younger age , absence of CHF , no preoperative use of beta blockers , and shorter cross clamp time .While the GenAlg-based models did not outperform STS prediction for patients with STS risk scores, our local-data-driven approach reliably predicted short LOS for cardiac surgery types that do not allow STS risk calculation. We advocate that each institution with sufficient observational data should build their own cardiac surgery risk models. With aging population, more and more geriatric patients with complex co-morbid conditions at increased risk for morbidity and mortality present for cardiac surgery . Risk scMorbidity is more common than mortality and usually leads to increased postoperative intensive care unit (ICU) length of stay (LOS) ,8. IncreTherefore, there is a need for a patient-level risk prediction model for all patients undergoing cardiac surgery, especially for those at a high risk such as geriatric patients. We hypothesized that a local, custom built model derived from a more homogeneous subset of patients would more accurately predict morbidity risk in geriatric populations, thus enabling proactive decision making. In the present study, we employed a machine learning technique called the genetic algorithm (GenAlg) to develop a customized model for short post-surgery LOS prediction and compared its performance with that of the STS score.We performed a retrospective medical records review of geriatric patients who were aged 70 or older, and underwent elective cardiac surgery from January 2002 to June 2011 at an urban tertiary-care center. We obtained approval from the Institutional Review Board of our institution. We obtained the records from the Society of Thoracic Surgeons (STS) database maintained by trained personnel at our institution since 2001. Two trained cardiac surgical nurses report outcomes to the STS database in a quarterly fashion. They have periodic internal checks and also receive reports from the Department of Public Health liaison on a yearly basis commenting on data quality and necessary corrections. The database contains all preoperative , intraoperative , and postoperative adverse outcomes .The STS predefined morbid events include :Operative mortality: death during the same hospitalization as surgery regardless of timing, or within 30 days of surgery regardless of venuePermanent stroke (cerebrovascular accident): a central neurologic deficit persisting longer than 72 hoursRenal failure: a new requirement for dialysis or an increase in serum creatinine to greater than 2.0 mg/dL and double the most recent preoperative creatinine levelProlonged ventilation: ventilation for more than 24 hoursDeep sternal wound infectionReoperation for any reasonMajor morbidity or mortality: a composite defined as the occurrence of any of the above end pointsProlonged postoperative LOS: LOS greater than 14 days Short postoperative LOS: LOS less than 6 days and patient discharged aliveThe following 22 preoperative features (predictors) were investigated for their predictive power for short LOS (defined by STS as LOS < 6 days): age, gender, race, family history of coronary artery disease (CAD), diabetes mellitus (DM), hypertension (HTN), chronic lung disease, cerebrovascular disease (CVD), peripheral vascular disease (PVD), myocardial infarction (MI), congestive heart failure (CHF), preoperative use of the following medications: beta blockers, angiotensin converting enzyme inhibitors (ACEI), intravenous (IV) nitrates, anticoagulants, inotropes, steroids, and aspirin; the type of cardiac surgery: coronary artery bypass graft placement (CABG), aortic valve procedure, mitral valve procedure, and last creatinine measurement prior to the surgery. In addition, one intraoperative variable was added to the feature pool: cross clamp time. Hence, a total of 23 features were investigated.We applied a GenAlg  to compuThe GenAlg was executed separately for varying numbers of selected features: 5, 10, 15, and 20. All features, 23 of them, were also evaluated without the GenAlg. Since GenAlg performance depends on the initial population of randomly selected feature sets, the GenAlg was repeated 5 times for each number of features. Among the 5 attempts, the feature combination corresponding to the highest AUC based on the validation data was selected for the final evaluation on the test data. Each GenAlg run was terminated after 50 generations.STS prediction for short LOS was not available for all patients in our cohort. This is due to the fact that STS does not have a prediction model for every type of cardiac surgery. Therefore, the GenAlg was conducted independently on two sub-cohorts: the patients with an STS short LOS prediction and those without. STS prediction served as the gold standard in the sub-cohort where STS prediction was available, and STS AUC was computed on the same test data on which the GenAlgselected features were evaluated.Lastly, we built an LR model on all patients  using the selected features in order to compute odds ratios (ORs) and the corresponding 95% confidence intervals (CI). However, this analysis was still conducted separately for the two sub-cohorts with and without STS prediction. Statistical significance was reached when p-value was less than 0.05.All feature selection and evaluation were conducted in MATLAB version R2010b .1426 patients met the inclusion criteria and were analyzed. Our objective was to assess whether a custom model would more accurately predict morbidity in comparison with the established STS model in our local geriatric population. We utilized a GenAlg, an evolutionary algorithm that can perform automated feature selection to maximize predictive performance. In addition to the preoperative variables already utilized in the STS score, we also used intraoperative variables as these variables also influence prognosis . In the Accurate prediction of prolonged post-cardiac-surgery LOS can be a crucial piece of information for health care cost savings. The earlier clinicians and hospital administrators are informed of potential excessive consumption of hospital resources, the better they can allocate the limited resources they have. Moreover, keeping LOS at minimum is important with respect to managing risk for hospital-acquired infections.Building customized models rather than the traditional one-model-fits-all approach has been shown to be meritorious in mortality prediction . Such taet al. [In a systematic review and validation of prediction of prolonged LOS following cardiac surgery, Ettema et al.  found thet al.   at other institutions for external validity. Ultimately, an impact study will have to be conducted to gauge the benefits of having accurate LOS prediction for cardiac patients with respect to cost savings and reduction of hospital-acquired infections.Our GenAlg-based models did not outperform STS prediction for patients with STS risk scores. However, our customized approach based on local data reliably predicted short LOS for cardiac surgery types that do not allow STS risk calculation. The primary strength of our proposed risk stratification is its utilization of the most relevant data from a local data repository rather than one-size-fits-all models. We advocate that each institution with sufficient observational data should build their own risk models.


Dengue virus (DV) infections cause undisputedly the most important arthropod-borne viral disease in terms of worldwide prevalence, human suffering, and cost. Worldwide DV infection prevalence in 2010 was between 284 to 528 million cases While up to 90% of dengue cases are either asymptomatic or cause an underreported, self-limited, flu-like illness, symptomatic DV infection manifests as two main clinical forms: 1) dengue fever (DF) and 2) severe dengue (SD). DF symptoms include high-degree fever, headache, myalgia with arthralgia, retro-orbital pain, and rash. SD evolves clinically as a life-threatening complication from DF. SD is characterized by plasma leakage, hemoconcentration, hemorrhagic shock, and organ failure, which may lead to patient death. Any of the four described DV serotypes can be responsible for either DF or SD, although the precise mechanisms leading to SD are still unclear. Evidence suggests factors such as viral virulence, host gene background, and secondary DV infections contribute to the exacerbation of the disease. In consequence, preventive strategies through vaccination must consider tetravalent formulations.Flaviridae family, of which DV is a member, consists of enveloped, positive-stranded RNA viruses. The flavivirus genome encodes a polyprotein, which is processed by cellular and viral-encoded proteases into ten proteins  and the attenuated strain 14-14-2 against Japanese encephalitis virus (JEV), both obtained by serial passage in cell culture. Importantly, for both vaccines, a neutralization titer of at least 1∶10 is protective. Thus, the most useful and documented correlate of protection for flaviviruses is the presence of neutralizing antibodies assayed by the plaque reduction neutralization assay (PRNA).Why then has the development of a DV vaccine proven so challenging? Natural DV infection triggers a robust, neutralizing immunity that provides an apparently life-long protection against the infecting DV serotype and a short-lived (months) cross-protection against heterologous DV serotypes. Interestingly, the humoral response to DV not only mediates protection though viral neutralization, but also seems to play a major role in the development of more severe forms of dengue disease. Dengue hemorrhagic fever and dengue shock syndrome (DHF/DSS) cases are often associated with secondary DV infections with a heterologous DV serotype. Subneutralizing, heterotypic antibodies induced during a primary infection with one serotype of DV form complexes with a different serotype of DV during secondary infection, enhancing uptake of infectious particles by Fc-receptor–bearing permissive cells. This phenomenon, called antibody-dependent enhancement (ADE), results in an increased number of infected cells and a higher viremia that, combined with a pro-inflammatory cytokine response from an enhanced infection of macrophage and dendritic cells, have been related with SD Attenuating DV is a quest that has lasted more than 60 years As an alternative to attenuation by serial passages, DV can also be attenuated by targeted mutagenesis. Deletion of 30 nucleotides (Δ30) in the 3’ end of the DV1 and DV4 RNA genome results in a reduction of viral replication fitness while maintaining immunogenicity A major breakthrough in DV vaccine research came with the demonstration that mosquito-borne flaviviruses may replicate as chimeric viruses in which the envelope genes M and E of an attenuated vector genome are replaced with the cognate genes of the target virus. This strategy, depicted in Using the replication machinery of the YFV 17D vaccine strain, Chimerivax displays each of the four DV serotype glycoproteins in a separate functional viral particle. It constitutes the most advanced vaccine candidate to be tested in humans so far. This vaccine showed promising results in phase I clinical trials in endemic and nonendemic countries. However a phase IIb clinical trial in 2,669 Thai schoolchildren with a tetravalent vaccine showed poor efficacy  of protection against febrile DV infections ClinicalTrial.gov identifier NCT01224639).The other chimeric construction approach uses the replication machinery of the aforementioned attenuated DV2 PDK53 vaccine candidate. Again, the relevant DV envelope glycoproteins are displayed on the surface of the chimeric particle. The tetravalent formulation (DENvax) is currently in a dose-escalating, phase I clinical trial  candidate have been documented ClinicalTrial.gov identifier NCT00936429).Subunit vaccines have also been explored. Immunization with an insect-expressed E protein ectodomain from DV2 is capable of inducing protection in macaques In summary, the strength and longevity of neutralizing immune responses elicited by attenuated vaccines outperforms responses observed with replication-defective approaches. Still, safety and the possibility of combining vaccine components without the possibility of viral interference adds enthusiasm for the latter approach. Interestingly, the potential for a prime-boost schedule was assessed in primate model bona fide correlate of protection because of ambiguous results pertaining to the protective titer. Testing the protection efficiency of tetravalent vaccine candidates in volunteers may answer both questions. Results from the first human DV challenge experiments have been recently published and demonstrate the viability of this approach There are two main windows of opportunity to build upon toward realizing efficient vaccination for DV. First, a clinically relevant animal model for dengue infection and vaccine development is lacking. Rhesus monkeys do not show clinical signs of infection after a wild-type DV challenge; instead the intensity and length of viremia serves as a proxy to infer protection. Second, rigorous correlates of protection have not been established for DV. The best available indicator of immunogenicity is the titration of neutralizing antibodies, however titration of plaque reduction neutralization antibodies has not been promoted to a


Hemodynamic mismatch responses can be elicited by deviant stimuli in a sequence of standard stimuli even during cognitive demanding tasks. Emotional context is known to modulate lateralized processing. Right-hemispheric negative emotion processing may bias attention to the right and enhance processing of right-ear stimuli. The present study examined the influence of induced mood on lateralized pre-attentive auditory processing of dichotic stimuli using functional magnetic resonance imaging (fMRI). Faces expressing emotions  were presented in a blocked design while a dichotic oddball sequence with consonant-vowel (CV) syllables in an event-related design was simultaneously administered. Twenty healthy participants were instructed to feel the emotion perceived on the images and to ignore the syllables. Deviant sounds reliably activated bilateral auditory cortices and confirmed attention effects by modulation of visual activity. Sad mood induction activated visual, limbic and right prefrontal areas. A lateralization effect of emotion-attention interaction was reflected in a stronger response to right-ear deviants in the right auditory cortex during sad mood. This imbalance of resources may be a neurophysiological correlate of laterality in sad mood and depression. Conceivably, the compensatory right-hemispheric enhancement of resources elicits increased ipsilateral processing. Laterality effects can emerge as a function of emotional state Laterality effects have been observed in affective disorders and related to the processing of emotion lateralized auditory stimuli in the healthy brain, however, are largely unknown. We hypothesized that ongoing emotion processing in frontal cortices can elicit an imbalance of processing resources with right auditory cortex showing reduced activation, resulting in enhanced processing of deviant sounds at the right ear.Several studies have investigated the effects of emotional context on pre-attentive processing of auditory stimuli with electrophysiological measures The present study investigated the influence of induced mood on laterality in the processing of neutral language stimuli. Using a design that elicited the hemodynamic analogue of the mismatch negativity (MMN), the influence of different mood states on the processing of unattended dichotically presented acoustic stimuli at auditory cortices was examined. Specifically, mood induction in healthy volunteers was achieved by showing emotional facial expressions  Twenty healthy volunteers (age 20–32 years) participated in the study. All subjects were right-handed, as indicated by the laterality quotient (mean 88.2±13.5) of the Edinburgh Inventory Auditory stimuli were consonant-vowel syllables /ba/, /da/, /ga/, /ka/, /pa/, and /ta/, each recorded twice to allow for a stereo effect when composing the dichotic stimuli, i.e., even the stimuli with the same syllable presented to each ear were not perceived as ‘inside the head’ For mood induction, 72 color photographs of actors expressing sadness, happiness or neutral emotion from a standardized stimulus set were selected A dichotic listening pretest for determining laterality was conducted outside the fMRI scanner. Every dichotic combination of the six different syllables /ba/, /da/, /ga/, /ka/, /pa/, and /ta/ (6×6 = 36) was presented ten times. Of these 360 stimuli, 300 items represented dichotic pairs composed of two different syllables. Subjects were asked to identify the most salient percept of each dichotic pair and indicated their answer in written form by choosing one of the six syllables in a 6-alternatives forced-choice task.Mood induction was carried out in a blocked fMRI design . Sad mooTwo types of mood ratings were applied After the last rating, a visuospatial attention task was presented. A small circle was randomly presented to the left or right visual field with a stimulus duration of 100 ms, a fixation duration of 800 ms, and a variable inter-trial interval of 2,500–4,000 ms. Subjects were instructed to fixate on the cross in the middle of the screen and to press a button as fast as possible when the circle appeared. Due to technical limitations, button responses were not recorded. The task was administered in order to involve bilateral attention resources and, as such, to ‘wash-out’ mood state and attention shifts prior to the next run. For fMRI, this time period was modeled as a nuisance variable.A task-irrelevant oddball sequence with dichotic stimuli was administered simultaneously to the mood induction run, with a stimulus onset asynchrony of 667 ms . The auditory oddball sequence consisted of frequent CV syllable /ba/ (different types of /ba/-recordings at the left and right channels: /baL/-/baR/). In the 10% deviants , the contralateral /ba/ items were identical within the respective channel of the frequent stimuli . Subjects were instructed to ignore the sounds and only pay attention to the mood induction procedure. This experimental design allowed examining the allocation of bottom-up controlled spatial attention without interfering with the mood induction task. Three different oddball sequences were prepared in advance and one assigned to each of the three mood conditions. This assignment was kept for the entire experiment. With the fourth image acquisition after three dummy scans, the oddball sequence started and lasted until the last image acquisition of the run.2, voxel size = 3×3 mm2). Slices covered the entire cerebral cortex and were positioned oblique-transversally to achieve maximal brain coverage. Two hundred and thirty volumes were collected per session. The first three volumes of each session were excluded to remove the influence of T1 saturation effects. Head movement was minimized with the use of foam wedges to securely hold the head in the 12-channel head coil. Structural images were obtained using a high-resolution T1-weighted 3-D sequence .Scanning was performed on a 3 T Magnetom Trio MR scanner  in the department of Psychiatry, Psychotherapy and Psychosomatics at the Medical School of the RWTH Aachen University. Functional images were collected with echo planar imaging (EPI) sensitive to blood oxygenation level dependent (BOLD) contrast . Earplugs further reduced scanner noise. The volume of the auditory stimuli was individually adjusted to a comfortable listening level and good audibility during scanner noise.A laterality index was computed by subtracting left-ear decisions from right-ear decisions in the 300 pairs of lexically different syllables . Laterality indices larger than zero indicated a right-ear advantage (REA). A group mean of right-ear minus left-ear decisions was computed.SAM ratings were analyzed for arousal and valence separately, with repeated-measures ANOVAs conducted with the factors mood and timepoint of rating (3×4). Significance level was set at p<.05 and then Bonferroni-corrected in pairwise comparisons. Ratings of the ESR were analyzed on a descriptive level www.fil.ion.ucl.ac.uk) implemented in MATLAB . After discarding the first three volumes, 227 volumes from each participant were spatially realigned to the mean image to correct for head movement. The next step was normalization into the stereotaxic anatomical MNI  space with 2 mm isotropic voxels. The normalized data were spatially smoothed with an 8 mm isotropic Gaussian kernel to account for inter-subject variability in brain anatomy and to increase signal-to-noise ratio.fMRI data analyses were calculated using Statistical Parametric Mapping software  and its temporal derivative for a more differentiated modeling of the time course. The design comprised six sessions  with three mood induction blocks per session and the deviant syllables as events. The following regressors were modeled for each session: instructions, ratings, mood induction blocks, visuospatial task, deviant syllable /ga/ presented to the left ear, deviant syllable /pa/ presented to the left ear, deviant syllable /ga/ presented to the right ear, deviant syllable /pa/ presented to the right ear.Contrast images at the individual level were computed comparing the three mood induction blocks in each session to the baseline based on the hrf (t-contrast). The three contrast images of the three mood induction conditions were each analyzed with a one-sample t-test at the group level. Only clusters above the cluster-level threshold according to FWE-corrected p<.05  were reported.At the individual level, contrast images were computed according to the factors mood , presentation side of deviant syllable  and BOLD response , resulting in twelve images per subject. To measure the effects of induced mood and side of deviant on brain activity, a repeated-measures model with condition as the fixed factor and subject as the random factor was applied and the twelve contrast images were implemented as conditions. Inference statistics were based on the effects of interest contrast (F-contrast) at the threshold of FWE-correction (p<.05). Only clusters with a minimal volume of 120 µl (15 voxels) were considered.mood  and presentation side of deviant  was conducted. Post-hoc testing disentangled the effects in pair-wise comparisons. To control for sex effects, an ANOVA was computed with the same design but including the intersubject factor gender. Significance level was set at p<.05 for hypothesis testing.For the hypothesis-driven region of interest (ROI) analyses, contrast estimates were extracted from the activation peaks in bilateral auditory cortices. To adjust the hemodynamic response, the generic model function and its derivative were weighted according to the best fit across all deviants. A repeated-measures ANOVA with the factors All subjects except one showed a higher right-ear than left-ear score, demonstrating an REA . Left hemisphere dominance for phonetic processing and language can therefore be assumed in this group.Mood induction was confirmed by significant mood effects on the arousal and valence ratings . The repValence ratings revealed a significant effect of mood  and a significant mood x time interaction . Pairwise comparisons (mean difference±SE) showed a significant effect of all pairs of mood conditions: sad and neutral , happy and neutral , as well as sad and happy  and the 2-level factor presentation side (left-ear vs. right-ear deviant) assessed the response amplitudes at the activation peaks at the left and right auditory cortices. At the left auditory cortex, a significant effect emerged only for presentation side  but not for mood . The interaction just failed significance . Post-hoc t-tests confirmed larger responses to right-ear deviant syllables in sad and happy mood  but not in the neutral condition  and presentation side of deviant . Importantly, a significant interaction of mood and presentation side emerged . As concerns the post-hoc t-tests for presentation side, the same pattern emerged as at the left hemisphere  reflecting a subtle modulation of variance by the introduced covariate. No significant main effect or interaction with the other predictors emerged .The present study examined the influence of induced mood on lateralized processing of acoustic stimuli in the auditory cortex in a group of left-hemisphere dominant healthy volunteers. During task-irrelevant dichotic stimulation, mood induction with emotional facial expressions yielded behavioral effects and activation in brain areas known to be involved in emotion processing, such as amygdala, hippocampus and prefrontal cortex. The phonetic deviants elicited hemodynamic mismatch responses in auditory and visual cortices. A strong modulation of lateralized processing by induced mood was observed. Both emotion conditions (sad and happy mood) yielded a relative preponderance of activation to right-ear deviants in left auditory cortex, whereas in right auditory cortex right-ear deviants elicited higher activation during sad mood, reflecting the interaction of negative emotion processing in the right hemisphere and lateralized auditory processing. Overall, deviant events presented to the right ear elicited strongest activation during sad mood.Consistent with previous studies, the present results indicate the success of mood induction Deviant events triggered activation in bilateral auditory and visual cortices. Activation of the auditory cortex to non-attended changes in the auditory stream is well established using electroencephalography EEG: , magnetoThe activated voxels in the visual cortex survived the rather conservative FWE-correction – though exhibiting a smaller effect size than auditory cortex. The mood induction task in the present study has an explicit visual component that requires participants to direct their attention towards the stimuli. Even though more standard MMN paradigms involve reading, watching a movie or even attending to a visual task e.g. , the preThe present study investigated how lateralized processing of deviant events is modulated by mood induction. Auditory responses to right-ear deviants were enhanced during sad mood as compared to neutral mood, reflecting mood-dependent modulation of mismatch responses to consonant-vowel syllables. Previous studies investigating the effect of emotional context on the processing of neutral auditory stimuli reported enhanced acoustic novelty processing for negative valence The mood induction procedure in the present study yielded similar arousal ratings for happy and sad mood, which were significantly higher than during the neutral condition  – in contask-irrelevant dichotic stimuli indicated increased processing of right-ear stimuli on the neural level as well. Liotti and Mayberg activation of prefrontal cortex instead of deactivation in the right hemisphere in sad mood as well as increased activation of right auditory cortex to ipsilateral deviants. This right-hemisphere overactivation may serve as a compensatory mechanism to reduce functional impairment of the right hemisphere in depression (for a review, see An increased REA, which is an explicit measure of laterality, was found in depressive patients Differences in the procedure may account for some discrepancy to a study that yielded a decreased REA after negative mood induction. Gadea et al. Subjects were not asked to report strategies for reaching the mood state shown in the emotional facial expressions. Subjects were only instructed to try to feel the mood seen on the pictures. One might argue that watching facial expressions of emotions only elicits perceptual processing of emotion rather than feeling the emotion. Mood ratings, however, indicated that arousal and valence changed according to the emotion presented and thus successful mood induction can be assumed. Ideally, an independent measure of mood would have been optimal to rule out any social desirability effect Whereas functional magnetic resonance imaging provides high spatial resolution, the temporal resolution is very poor compared to electrophysiological measures. As such, we cannot draw conclusions about latency and duration effects of the auditory cortex responses. On the other hand, without the higher spatial resolution in fMRI, we would not have been able to reveal the visual cortex activation to deviant events. Given that the laterality of mismatch responses to auditory stimuli follows a time course Gender effects in emotion processing (see for example The present study demonstrated an influence of induced mood states on auditory cortex activation to dichotically presented deviant syllables. Prefrontal top-down influences on auditory processing may be the underlying mechanism leading to enhanced processing of right-ear deviants. Moreover, compensatory resource allocation to the right hemisphere in the context of negative valence was reflected in higher excitability of the right auditory cortex to ipsilateral deviants. Present findings emphasize the role of cognitive and emotion lateralization by indicating a strong mood-dependent modulation of lateralized auditory processing, even in the absence of voluntarily directed attention to spatially presented stimuli. Ipsilateral processing may account for the enhanced right-hemispheric response to right-ear deviants during mood changes.


The Fanconi Anemia (FA) pathway consists of proteins involved in repairing DNA damage, including interstrand cross-links (ICLs). The pathway contains an upstream multiprotein core complex that mediates the monoubiquitylation of the FANCD2 and FANCI heterodimer, and a downstream pathway that converges with a larger network of proteins with roles in homologous recombination and other DNA repair pathways. Selective killing of cancer cells with an intact FA pathway but deficient in certain other DNA repair pathways is an emerging approach to tailored cancer therapy. Inhibiting the FA pathway becomes selectively lethal when certain repair genes are defective, such as the checkpoint kinase ATM. Inhibiting the FA pathway in ATM deficient cells can be achieved with small molecule inhibitors, suggesting that new cancer therapeutics could be developed by identifying FA pathway inhibitors to treat cancers that contain defects that are synthetic lethal with FA. Fanconi anemia is a rare genetic disease featuring characteristic developmental abnormalities, a progressive pancytopenia, genomic instability, and predisposition to cancer , 2. The  Although FA is rare, understanding the functional role of the FA proteins in context with other DNA damage response pathways will provide broader opportunities for new cancer therapeutics. Two general strategies could accomplish this, as illustrated in A defining characteristic of FA cells is hypersensitivity to cross-linking agents, such as the chemotherapeutic agent cisplatin , 5. CispIn addition to sensitization, inhibiting the FA pathway may be an effective strategy to exploit synthetic lethal interactions aimed at improving targeted killing of tumor cells. Current approaches in cancer treatment are generally not selective, affecting both cancer cells and normal cells. However, inactivation of DNA repair pathways, an event that occurs frequently during tumor development , can makDefects in HR repair can result in an overreliance on the protein PARP1, which is responsible for repair of DNA single strand breaks by the base excision repair pathway. Unrepaired single-strand breaks are converted to double-strand breaks during replication and must be repaired by HR –27. Thusin vitro and to increase radiosensitivity of other ATM-proficient cell lines, including nonsmall-cell lung cancer, medulloblastoma, ependymoma, and high-grade gliomas [PARP1 Inhibition has also been shown to be selectively toxic to ATM-defective tumor cell lines  gliomas –35. In a gliomas , 37.PTEN (phosphatase and tensin homolog) is a tumor-suppressor gene and one of the most commonly mutated genes in human tumor cells , 39 see. PTEN deSynthetic lethal interactions with the FA pathway have been explored. An siRNA-based screen of cells deficient in the Fanconi core complex protein, FANCG, showed that ATM, PARP1, NBS1, and PLK1 were among the genes with a synthetic lethal interaction  see Tab. The FA-Xenopus egg extracts to screen small molecules for stronger and more specific inhibitors of FANCD2 monubiquitylation. Unlike cell-based screening assays for small molecules capable of inhibiting the FA pathway, the cell-free method uncouples FANCD2 monoubiquitylation from DNA replication, thus focusing more specifically on the key biochemical steps in a soluble context enriched for nuclear proteins and capable of full genomic replication [Inhibition of the FA pathway could occur at any point in the multistep FA protein network, but a key predictive readout for FA function and resistance to ICLs is the monoubiquitylation of FANCD2 , 55. Sevlication . Screenilication , 57. A slication , 59. Thelication , 57. Thelication , 52. Understanding how the Fanconi anemia pathway functions in concert with other DNA damage response networks is essential for understanding genomic stability and for exploiting synthetic lethality for new cancer treatments. New chemotherapeutic agents could be developed by identifying potent and specific inhibitors of the FA pathway, for example, by screening for compounds that inhibit key FA pathway steps . While a long-term defect in the function of the FA pathway would result in genomic instability, short-term inhibition could provide a treatment strategy for tumors with deficiencies in certain other DNA repair pathways. Stringent identification of additional genes with synthetic lethal relationships with the FA pathway, and identification of malignancies with deficiencies or mutations in genes that are synthetic lethal with FA will be required for these tailored therapeutic approaches.