Praziquantel represents the frontline chemotherapy used to treat schistosomiasis, a neglected tropical disease (NTD) caused by infection with macro-parasitic blood fluke schistosomes. While this drug is safe, its inability to kill all schistosome lifecycle stages within the human host often requires repeat treatments. This limitation, amongst others, has led to the search for novel anti-schistosome replacement or combinatorial chemotherapies. Here, we describe a repositioning strategy to assess the anthelmintic activity of epigenetic probes/inhibitors obtained from the Structural Genomics Consortium.Schistosoma mansoni schistosomula using the high-throughput Roboworm platform. At 10 μM, 14 of these 37 compounds (38%) negatively affected schistosomula motility and phenotype after 72 hours of continuous co-incubation. Subsequent dose-response titrations against schistosomula and adult worms revealed epigenetic probes targeting one reader (NVS-CECR2-1), one writer (LLY-507 and BAY-598) and one eraser (GSK-J4) to be particularly active. As LLY-507/BAY-598 (SMYD2 histone methyltransferase inhibitors) and GSK-J4 (a JMJD3 histone demethylase inhibitor) regulate an epigenetic process (protein methylation) known to be critical for schistosome development, further characterisation of these compounds/putative targets was performed. RNA interference (RNAi) of one putative LLY-507/BAY-598 S. mansoni target (Smp_000700) in adult worms replicated the compound-mediated motility and egg production defects. Furthermore, H3K36me2, a known product catalysed by SMYD2 activity, was also reduced by LLY-507 (25%), BAY-598 (23%) and siSmp_000700 (15%) treatment of adult worms. Oviposition and packaging of vitelline cells into in vitro laid eggs was also significantly affected by GSK-J4 (putative cell permeable prodrug inhibitor of Smp_034000), but not by the related structural analogue GSK-J1 (cell impermeable inhibitor).Thirty-seven epigenetic probes/inhibitors targeting histone readers, writers and erasers were initially screened against Collectively, these results provide further support for the development of next-generation drugs targeting schistosome epigenetic pathway components. In particular, the progression of histone methylation/demethylation modulators presents a tractable strategy for anti-schistosomal control. Schistosoma mansoni epigenetic processes, we performed anthelmintic screening of 37 epigenetic probes/epigenetic inhibitors obtained from the Structural Genomics Consortium (SGC). The results of these studies highlighted that schistosome protein methylation/demethylation processes are acutely vulnerable. In particular, compounds affecting schistosome SMYD  or JMJD (GSK-J4) homologues are especially active on schistosomula and adult worms during in vitro phenotypic drug screens. The active epigenetic probes identified here as well as their corresponding S. mansoni protein targets offers new starting points for the development of next-generation anti-schistosomals.Human schistosomiasis is caused by infection with parasitic blood fluke worms. Global control of this NTD is currently facilitated by administration of a single drug, praziquantel (PZQ). This mono-chemotherapeutic strategy of schistosomiasis control presents challenges as PZQ is not active against all human-dwelling schistosome lifecycle stages and the evolution of PZQ resistant parasites remains a threat. Therefore, new drugs to be used in combination with or in replacement of PZQ are urgently needed. Here, continuing our studies on  Amongst human infectious diseases caused by macro-parasitic organisms, schistosomiasis is the most significant in terms of its negative impact on both individual health and population-driven socio-economic outputs –3. The cOne recent approach applied to schistosome drug discovery is based on the concept of compound repositioning or repurposing, where new indications for existing drugs are sought . Two benS. mansoni) of thirty-four SGC-supplied EPs  and three EIs (compounds that do not display these specific epigenetic probe traits)  authors used such a small incubation time (20h) with cytotoxicity assays on the human cell line for calculating selectivity indexes. It's an issue since parasite assays were all performed with 72h incubation time. The authors should generate new data with 72h incubation or make a strong case justifying their choice for 20h since keeping the later should overestimated the selectivity/safety of compounds"Reviewer's Responses to QuestionsKey Review Criteria Required for Acceptance?As you describe the new analyses required for acceptance, please consider the following:Methods-Are the objectives of the study clearly articulated with a clear testable hypothesis stated?-Is the study design appropriate to address the stated objectives?-Is the population clearly described and appropriate for the hypothesis being tested?-Is the sample size sufficient to ensure adequate power to address the hypothesis being tested?-Were correct statistical analysis used to support conclusions?-Are there concerns about ethical or regulatory requirements being met?Reviewer #1: -Are the objectives of the study clearly articulated with a clear testable hypothesis stated?Yes-Is the study design appropriate to address the stated objectives?Yes-Is the population clearly described and appropriate for the hypothesis being tested?NA-Is the sample size sufficient to ensure adequate power to address the hypothesis being tested?NA-Were correct statistical analysis used to support conclusions?Yes-Are there concerns about ethical or regulatory requirements being met?NAReviewer #2: Several minor details in the methods text could be clarified. - Line 149. What strain of Mus musculus?- Line 188. Schistosomules were cultured with compound at 37 degrees – were they also imaged at 37 or at room temperature?- Line 189. Reference 50  is mentioned regarding phenotype and motility. However, some details on this package could be clarified. Is this package publically available? What was the rational for the hit thresholds (dotted line figure 1A and 1B)?- For RNAi, what concetration of siRNA duplexes were used?- Line 263. How many worms sample input are used in this assay?Reviewer #3: Methods are appropriate for the study--------------------Results-Does the analysis presented match the analysis plan?-Are the results clearly and completely presented?-Are the figures  of sufficient quality for clarity?Reviewer #1: -Does the analysis presented match the analysis plan?Yes-Are the results clearly and completely presented?Mostly.-Are the figures  of sufficient quality for clarity?Yes.Reviewer #2: The results are clearly presented. As mentioned above in the methods, a description of the hit threshold scores shown in Figure 1A and 1B could be elaborated upon. Either on the legend, or in the text around line 361-364 when these cutoffs are mentioned.For table 2, add in either the table or the legend a description of the assay used to generate the IC50 data for adult males and females. Also, in the text around line 409 where these results are discussed.The structures of the chemicals shown in table 2 should be included. It may not be convenient to include them in the actual table 2, but perhaps a supplemental table or in text form . While this is not essential, I am interested in whether pharmacological or genetic inhibition of histone methylation effects mitotic activity of somatic neoblasts or germ tissues ? Is this the mechanism by which egg laying is arrested?Reviewer #3: Results match the analysis plan.--------------------Conclusions-Are the conclusions supported by the data presented?-Are the limitations of analysis clearly described?-Do the authors discuss how these data can be helpful to advance our understanding of the topic under study?-Is public health relevance addressed?Reviewer #1: -Are the conclusions supported by the data presented?Generally, yes.-Are the limitations of analysis clearly described?Mostly.-Do the authors discuss how these data can be helpful to advance our understanding of the topic under study?Yes.-Is public health relevance addressed?Yes.Reviewer #2: The conclusions are well supported by the data. Some discussion could elaborate the repurposing of epigenetic probes / inhibitor compounds. As mentioned in the introduction, praziquantel is not effective against juvenile life stages . Granted, ex vivo assays on these worms are more challenging – but is anything known about the expression of targets such as Smp_007000 across the schistosome life cycle? Given the effects on egg laying, it would be interesting whether these are differentially expressed in males and females or in the gonads (PMID: 27499125).Reviewer #3: Conclusions are appropriate.--------------------Editorial and Data Presentation Modifications?Use this section for editorial suggestions as well as relatively minor modifications of existing data that would enhance clarity. If the only modifications needed are minor and/or editorial, you may wish to recommend “Minor Revision” or “Accept”. Reviewer #1: (No Response)Reviewer #2: (No Response)Reviewer #3: minor revision--------------------Summary and General CommentsUse this section to provide overall comments, discuss strengths/weaknesses of the study, novelty, significance, general execution and scholarship. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. If requesting major revision, please articulate the new experiments that are needed.Reviewer #1: The authors describe the anti-schistosomal activities of compounds that are known modulators/inhibitors of human molecular targets involved in epigenetic processes. The deck of 37 compounds was obtained from the Structural Genomics Consortium and tested against schistosomula and adult schistosome lifecycle stages. The study combines several approaches to support/validate the main hits from the initial screenings: in silico modelling, RNAi-mediated functional genomics, enzyme assays and confocal microscopy. This is an important work with interesting findings to schistosomal drug discovery targeting epigenetic processes. Hence, it should be highly appreciated by the journal readership. I list below several minor points to be corrected but the only major point to be addressed is why authors used such a small incubation time (20h) with cytotoxicity assays on the human cell line for calculating selectivity indexes. It's an issue since parasite assays were all performed with 72h incubation time. The authors should generate new data with 72h incubation or make a strong case justifying their choice for 20h since keeping the later should overestimated the selectivity/safety of compounds.Minor points:- Methods:1- Compound acquisition, storage and handling: Please indicate where appropriate that compounds structures can be found at Table S12- Homology modelling and epi-drug docking: 2a - Provide  evidence for model validation and quality, having special attention to any issues involved in the proposed docking site.2b - Are all the EP/Eis know to be competitive with the substrates? if so, authors should state it earlier.- Results:Lines 311-313: Kd, Ki or IC50? the term potency may be misleading since it's usually expressed as an inverse proportional term. Then, higher potency may actually mean < 100nm.="" lines="" _348-3493a_="" figure="" 1="" _legend3a_="" please="" indicate="" if="" dashes="" crossing="" vertical="" connecting="" each="" replicate="" mean="" averages="" or="" medians.="" line="" _3853a_="" _please2c_="" complement="" s1="" by="" including="" chemical="" structures="" for="" the="" compounds.="" _386-3873a_="" authors="" contend="" fact="" that="" none="" of="" 386="" these="" chemotype="" controls="" demonstrated="" anti-schistosomula="" activity="" suggests="" specificity="" hits="" s.="" mansoni="" homologue="" is="" similar="" to="" found="" original="" human="" target="" but="" other="" interpretation="" would="" be="" they="" lack="" proper="" sar="" targets.="" it="" should="" discussed.="" table="" _23a_="" all="" ec50="" and="" cc50="" accompanied="" standard="" errors="" _9525_="" confidence="" intervals.="" also="" incubation="" time="" below="" headings.="" in="" _footnote2c_="" define="" _22_accurate22_.="" _413-4153a_="" are="" there="" any="" discussed="" compounds="" lly-507="" _bay-5983f_="" _4443a_="" _22_strong22_="" _22_close22_="" _463-4783a_="" make="" their="" conclusions="" lighter="" using="" expressions="" like="" _22_is="" predicted="" _bind...22_="" _22_predicted="" interaction="" may="" _prevent...22_="" _5233a_="" clarify="" why="" gsk-j4="" a="" prodrug.="" how="" drug="" _processed3f_="" _typos3a_="" _713a_="" theat="" threat="" _1273a_="" _22_cultured="" _this22_="" with="" _4523a_="" _22_ceb2_="" _stands22_="" _strands22_="" reviewer="" _23_23a_="" this="" study="" screened="" panel="" known="" epigenetic="" against="" schistosomules="" then="" proceeded="" active="" more="" detail="" on="" adult="" parasites="" _e28093_="" profiling="" phenotypic="" outcomes="" such="" as="" motility="" egg="" _laying2c_="" well="" assaying="" histone="" methylation.="" pharmacological="" experiments="" were="" controlled="" negative="" further="" validated="" rnai="" data="" showing="" knockdown="" putative="" methyltransferase="" phenocopies="" treatment.="" convincing="" controlled.="" laying="" phenotype="" particular="" _promising2c_="" since="" pathology="" associated="" disease="" driven="" parasite="" eggs.="" several="" suggestions="" clarification="" methods="" results="" have="" been="" outlined.="" _23_33a_="" manuscript="" written="" will="" interest="" pntd="" readers.="" minor="" revisions="" suggested.="" abstract="" author="" summary="" _numerous2c_="" undefined="" abbreviations.="" distracting="" _confusing3a_="" _482c_="" _532c_="" 76.="" _general2c_="" has="" large="" number="" _abbreviations2c_="" making="" tedious="" _read2c_="" do="" not="" addressed.="" 54="" _1525_="" something="" indicated.="" define.="" at="" places="" _e2809c_cell="" permeable="" _prodruge2809d_="" described.="" _however2c_="" _e2809c_worm="" _permeablee2809d_="" another="" matter="" general="" poorly="" understood.="" from="" overstated="" absence="" direct="" enzymatic="" evidence="" inhibition.="" was="" done="" indirectly="" h3k36me2="" levels="" _studies2c_="" which="" better="" support="" inhibitor="" _link2f_mechanism="" action.="" 311="" 313="" /> signs should be reversed.A general comment: the reality of therapy for schistosomiasis is that compounds need to be active after a single dose, where active means worm killing. Blocking motility or egg production after extended exposure to a compound will not be adequate for novel therapies. Did the compounds tested actually _kill_ the worms?--------------------PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.If you choose “no”, your identity will remain anonymous but your review may still be made public.Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.Reviewer #1: Yes: Floriano Paes Silva-JrReviewer #2: NoReviewer #3: No 28 Oct 2019AttachmentResponse to reviewers_Whatley and Padalino.docxSubmitted filename: Click here for additional data file. 30 Oct 2019Dear Prof. Hoffmann,We are pleased to inform you that your manuscript, "The repositioning of epigenetic probes/inhibitors identifies new anti-schistosomal lead compounds and chemotherapeutic targets", has been editorially accepted for publication at PLOS Neglected Tropical Diseases.Before your manuscript can be formally accepted and sent to production you will need to complete our formatting changes, which you will receive in a follow up email. 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If there is anything that the journal should know, please get in touch.* If you or your institution will be preparing press materials for this manuscript, please inform our press team in advance at http://www.editorialmanager.com/pntd, log in, and click on the "Update My Information" link at the top of the page. Please update your user information to ensure an efficient production and billing process.*Now that your manuscript has been provisionally accepted, please log into EM and update your profile. Go to http://www.plosntds.org/static/latexGuidelines.action] in the meantime.*Note to LaTeX users only - Our staff will ask you to upload a TEX file in addition to the PDF before the paper can be sent to typesetting, so please carefully review our Latex Guidelines [Thank you again for supporting open-access publishing; we are looking forward to publishing your work in PLOS Neglected Tropical Diseases.Best regards,Philip T. LoVerdeGuest EditorPLOS Neglected Tropical DiseasesJennifer KeiserDeputy EditorPLOS Neglected Tropical Diseases***********************************************************The authors have satisfactorily addressed the concerns of the reviewers. 7 Nov 2019Dear Prof. Hoffmann,We are delighted to inform you that your manuscript, "The repositioning of epigenetic probes/inhibitors identifies new anti-schistosomal lead compounds and chemotherapeutic targets," has been formally accepted for publication in PLOS Neglected Tropical Diseases.We have now passed your article onto the PLOS Production Department who will complete the rest of the publication process. All authors will receive a confirmation email upon publication.The corresponding author will soon be receiving a typeset proof for review, to ensure errors have not been introduced during production. Please review the PDF proof of your manuscript carefully, as this is the last chance to correct any scientific or type-setting errors. 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Coronary artery aneurysm is a rare condition which constitutes a small proportion of coronary artery disease. Such condition mimicking an intra-cardiac mass is extremely rare and poorly understood.We present an unusual case of a 53-year-old female with decreased exercise tolerance and lower extremity edema for 3 months. The echocardiography showed moderate tricuspid regurgitation and a right intra-ventricular mass below the tricuspid valve. No ventricular wall akinesia or ST segment change was found on echocardiography or electrocardiogram. Coronary computed tomographic angiography confirmed the diagnosis of intra-ventricular mass with feeding vessel originated from the right coronary artery. The patient was scheduled for tumor resection, and the mass turned out to be a thrombosed giant right coronary artery aneurysm. The patient received successful aneurysm resection and had an uneventful postoperative recovery. Unfortunately, a fistula between right coronary artery and right ventricle was detected on follow-up three months later by echocardiography.Coronary artery aneurysms presenting as intra-cardiac masses are extremely rare. Comprehensive preoperative evaluation is highly recommended because the surgical strategies for tumor and aneurysm are completely different. Aneurysm resection with bypass surgery is preferred rather than aneurysm repair. To our best knowledge, coronary artery aneurysms presenting as intra-ventricular masses have seldom been reported. Coronary artery aneurysm is a rare condition of coronary artery disease. It is defined as segmental coronary dilatation that exceeds the diameter of normal adjacent artery or the diameter of the largest artery by 1.5 times , with co9/L) and percentage (14.4%). Tumor markers were negative for malignancy. The transthoracic echocardiography (TTE) showed a cystic mass (3.5 × 2.7 cm) with hyperechoic external capsule locating below the tricuspid annulus  are of great necessity and importance for differential diagnosis between aneurysm and tumor. MRI is the most sensitive modality for characterizing the tumor tissue due to its excellent soft tissue contrast and variable manipulable parameters. In addition, coronary angiography is valuable to confirm diagnosis of coronary artery aneurysm and myocardial infarction, which is helpful for the surgical strategy.The management of coronary artery aneurysm includes medical intervention, stent insertion and surgical resection . IncidenWe experienced a rare case with a giant right coronary artery aneurysm mimicking a right intra-ventricular mass. MRI and coronary angiography are of great importance and value for preoperative diagnosis. We emphasize the necessity of comprehensive evaluation and accurate diagnosis before further treatment. For symptomatic patients with giant coronary artery aneurysms, aneurysm resection with bypass surgery is highly recommended.


The regulation of ubiquitination status in the cell is controlled by ubiquitin ligases acting in tandem with deubiquitinating enzymes. Ubiquitination controls many key processes in the cell from division to death making its tight regulation key to optimal cell function. Activity based protein profiling has emerged as a powerful technique to study these important enzymes. With around 100 deubiquitinating enzymes and 600 ubiquitin ligases in the human genome targeting a subclass of these enzymes or even a single enzyme is a compelling strategy to unpick this complex system. In this review we will discuss different approaches adopted, including activity-based probes centered around ubiquitin-protein, ubiquitin-peptide and mutated ubiquitin scaffolds. We examine challenges faced and opportunities presented to increase specificity in activity-based protein profiling of the ubiquitin conjugation/deconjugation machinery. Also, the presence of a native isopeptide bond in the scissile position may affect probe stability in complex systems.A further example of this probe type was developed by the Chatrerjee laboratory in 2016 using a selenocystine ligation  to sequentially functionalize thiols at a single carbon center for profiling E1 enzyme activity (Stanley et al., 73-thioester was reacted with azidoaminoethane to afford Ub-azide. Alkyne functionalized acrylate and acrylamide were used to prepare two TDAE functionalized ubiquitin monomers. A single cysteine mutant of His-UBE2L3 was reacted with the monomers to form the E2-Ub conjugate probes (Elaborated TDAE probes aimed to specifically target the E3 ligase Parkin by incorporating ubiquitin into the probe (Pao et al., e probes . The trie probes  later sh75-NH2NH2 using native chemical ligation followed by Dha formation. The probes were tested against catalytic domains of NEDD4 and UBE3C, labeling both active sites. Probing HeLa cells saw enrichment of several E3 enzymes, with strong enrichment of NEDD4. This work demonstrates an alternative route to E2-Ub probes however the linker is three atoms longer than the native and the electrophilic trap is presented two and five atoms away from Ub and UBE2D2, respectively.Shi and co-workers (Xu et al., Several approaches have been taken to confer selectivity to activity-based probes for ubiquitin conjugation/deconjugation machinery . MutatioCertain conjugate probes are limited due to their hydrolysable linkers and despite several well-designed solutions, many of the probes also do not perfectly mimic the linker length or trap position of the wild-type substrate. There is therefore scope for optimisation of the probe design and implementation of new chemistry for the synthesis of novel probes. Furthermore, these probes are currently limited by their lack of cell permeability. Recent work demonstrated that incorporation of cleavable cell-penetrating peptides can help deliver monoubiquitin probes into a cell (Gui et al., JM and NT drafted and edited the manuscript.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.


Posterior reversible encephalopathy syndrome (PRES) is a rare neurological disorder usually associated with specific medical conditions that cause a disturbance of the CNS homeostasis. It has seldom been reported to be a consequence of an iatrogenic intervention causing intracranial hypotension.We report the case of an individual 69-year-old male presenting with headache and blurred vision following cerebrospinal fluid (CSF) leak from resection of a sellar mass. The patient developed the condition following removal of the lumbar drain post-operatively. Magnetic Resonance Imaging showed bilateral occipital, parieto-occipital, and cerebellar T2 FLAIR hyper-intensities, suggesting a radiological diagnosis of posterior reversible encephalopathy syndrome (PRES). The patient’s symptoms started to improve shortly afterwards and had completely resolved at 3 months follow-up.The absence of severe hypertension and presence of an intraoperative CSF leak requiring placement of the lumbar drain suggests that decreased CSF volume and associated reactive hyperemia could have a role in the pathophysiology of the disease. Posterior reversible encephalopathy syndrome (PRES) originally described by Hinchey includes neurological abnormalities and associated characteristic findings on neuroimaging . PRES isOur patient is a 69-year-old Caucasian male with known hypertension, hyperlipidemia, gout and sialolithiasis presenting for endoscopic endonasal resection of a recurrent intra/suprasellar mass with cavernous sinus extension with pathological specimen consistent with Rathke’s cleft cyst. Cerebrospinal fluid (CSF) leak occurred intra-operatively requiring placement of a lumbar drain at a rate of 10 cc/h. CSF pressure was not elevated during the initial placement. Drainage continued intermittently at the same rate for 48 h. The drain was then clamped for 24 h prior to removal on post-operative day three without adverse event. The patient did not complain of any headaches following the surgical intervention but developed a severe positional headache associated with nausea, vomiting, and significantly blurred vision. The headaches were worsened upon sitting upright and partially relieved in the recumbent position. The ophthalmological examination demonstrated decreased visual acuity at 20/30 bilaterally. The optic disc retained spontaneous venous pulsations, suggestive of low or normal intracranial pressure.CT of the head revealed subcortical hypodensities at the posterior tip of the occipital lobes bilaterally. MRI demonstrated patchy areas of T2 and T2 FLAIR hyper-intensity in the bilateral posterior occipital lobes as well as the bilateral parieto-occipital junction and posterior cerebellum Fig. .Fig. 1AThe patient’s systolic blood pressure is charted in Fig. Posterior reversible encephalopathy syndrome is a rare disorder, and its mechanism remains controversial. It has been previously described occurring following lumbar CSF drainage –10. One Another potential mechanism of action of PRES involves endothelial dysfunction leading to an increase in membrane permeability and vasogenic edema as seen on imaging , 15. EveRecently, intracranial hypotension has also been associated with PRES . DemonstPRES is a potential consideration in post-operative patients with CSF leak with sudden neurological changes, specifically deficits in vision or seizures. The mechanism of PRES remains controversial and further investigations should be conducted whenever it does occur to advance our understanding of its etiology.


Pseudomonas syringae (Ps) are recognized as the most damaging factors in fruit trees with a significant economic and sanitary impact on crops. Among them, bacterial canker of apricot is exceedingly difficult to control due to a lack of efficient prophylactic measures. Several sources of partial resistance have been identified among genetic resources but the underlying genetic pattern has not been elucidated thus far. In this study, we phenotyped bacterial canker susceptibility in an apricot core-collection of 73 accessions over 4 years by measuring canker and superficial browning lengths issued from artificial inoculations in the orchard.Diseases caused by G) and genetic x year (G × Y) interaction effects extracted from linear mixed models.In order to investigate the genetic architecture of partial resistance, we performed a genome-wide association study using best linear unbiased predictors on genetic  between these two markers revealing an inter-chromosomal LD block linking the two underlying genes. This result supports the hypothesis of a co-adaptation effect due to selection through population demography. Candidate genes annotations suggest a functional pathway involving abscisic acid, a hormone mainly known for mediating abiotic stress responses but also reported as a potential factor in plant-pathogen interactions.We detected 11 significant associations over 7 candidate loci linked to disease resistance under the two most severe years. Colocalizations between Rosaceae species. It provided tools for fruit tree breeding by identifying progenitors with favorable haplotypes and by providing major-effect markers for a marker-assisted selection strategy.Our study contributed to the first detailed characterization of the genetic determinants of partial resistance to bacterial canker in a The online version of this article (10.1186/s12870-019-1631-3) contains supplementary material, which is available to authorized users. For decades, breeding programs have notably focused on the introgression of major resistant genes into crop cultivars. However multiple episodes of resistance breakdown have led research efforts to target polygenic  resistances . AlthougPrunus genus, apricot (Prunus armeniaca L.) is one of the most popular and typical crops of the Mediterranean Basin which provides 49% of the total production to the world market _[physical position in bp]. (XLSX 17724 kb)


Exploring virus–host interactions is key to understand mechanisms regulating the viral replicative cycle and any pathological outcomes associated with infection. Whereas interactions at the protein level are well explored, RNA interactions are less so. Novel sequencing methodologies have helped uncover the importance of RNA–protein and RNA–RNA interactions during infection. In addition to messenger RNAs (mRNAs), mammalian cells express a great number of regulatory non-coding RNAs, some of which are crucial for regulation of the immune system whereas others are utilized by viruses. It is thus becoming increasingly clear that RNA interactions play important roles for both sides in the arms race between virus and host. With the emerging field of RNA therapeutics, such interactions are promising antiviral targets. In this review, we discuss direct and indirect RNA interactions occurring between RNA viruses or retroviruses and host non-coding transcripts upon infection. In addition, we review RNA virus derived non-coding RNAs affecting immunological and metabolic pathways of the host cell typically to provide an advantage to the virus. The relatively few known examples of virus–host RNA interactions suggest that many more await discovery. Viruses are likely the most abundant biological entity and the known global virosphere is greatly expanding ,2. ViralHerpesviridae) or single-stranded  genomes; RNA viruses have double-stranded , or single-stranded positive sense  or negative sense  genomes, where orientation is defined for coding mRNA sequence to be of positive polarity. In addition, Hepadnaviridae are DNA viruses with RNA intermediates, and Retroviridae are (+)RNA viruses with DNA intermediates . U. U87]. U365 . . 164]. ICulex pipiens accumulate in midgut cells and display RNAi suppressor activity by counteracting Dicer. This process modulates transmission of the virus to the salivary glands and thereby its transmission to the next host [-/--IRF7-/- or IFNAR-/- mice with sfRNA defective WNV strains demonstrated that sfRNA production is critical to sustain infection in animals with intact interferon response [The biological functions of sfRNAs vary between organisms infected B. WNV deext host ,173. In ext host ,167,174.ext host . In addiresponse . An alteWhereas viral lncRNAs are well described for large DNA viruses like the PAN RNA expressed by Kaposi’s sarcoma-associated herpesvirus (KSHV) , lncRNAsIn recent years, there has been an increasing interest in developing tools to systematically dissect the biological significance of RNA interactions governing cellular processes as well as pathogen–host interactions. These novel methodologies are expanding our knowledge on gene expression regulation mediated by ncRNAs. It is also becoming clear that such mechanisms play central roles in the regulation of the cellular response to viral infection. Direct binding to cellular ncRNAs is further exploited by RNA viruses to their own advantage, and this could provide new modalities for therapy. Thus, virus–host RNA interactions are becoming a novel focus area to fully understand the complexity of the virus life cycle, but more systematic investigation is required. The role of cellular and viral-derived small RNA is further along compared to lncRNAs and other RNA species. Despite such progress, much remains to be investigated to understand which virus–host RNA interactions are the result of the cell attempting to clear the infection and which represent a viral strategy to create a favorable host cell environment. Lastly, the often narrow focus on pathogenic viral species suggests that a broader investigation of the RNA virosphere will reveal exciting discoveries in this emerging field.Unanswered questionsAre there undiscovered cases of RNA viruses benefiting from miRNA-mediated repression of replication in certain tissues?What undiscovered examples of direct viral RNA-miRNA interactions exist and what are their functions?Does virus-mediated RNA sequestration have functional consequences for pathology or immune responses?Is miRNA manipulation, even for direct use in genome replication, a feature of persistent viruses?Does non-coding RNA in post-transcriptional regulation play unappreciated roles during chronic infection?What other types of cellular RNA interact directly with viral RNA?Do other mammalian RNA viruses accumulate functional non-coding RNA species akin to sfRNAs?


SF3B1) and two splicing regulators bound to 3’ splice sites (U2AF1) or exonic splicing enhancers (SRSF2) are particularly prevalent and mutually exclusive [Recent genomic characterization of cancers has revealed recurrent somatic mutations in genes encoding RNA splicing factors. As many as 119 splicing factors were implicated in a recent pan-cancer analysis [e sites UAF1 or exe sites UAF1 or exGrowing evidence suggests that defective RNA processing can induce accumulation of R-loops, transcription intermediates consisting of RNA:DNA hybrids and displaced single-stranded DNA (ssDNA) . We prevIt is not yet known how the increase of R-loops impacts the cancer genome. R-loops are known to interfere with DNA replication and induce replication stress . FurtherU2AF1 mutations [Emerging evidence has also suggested that the normal function of ATR may be altered in cells with spliceosome mutations. First, alternatively spliced ATR transcripts were found in AML patients harboring utations . An indeutations . Interesutations . How theBased on these pre-clinical findings, we recently initiated a single-arm phase Ib trial to assess safety and seek preliminary evidence of efficacy for the ATR inhibitor, AZD6738, in patients with MDS and chronic myelomonocytic leukemia who have failed first line therapy (NCT03770429). We will test the hypothesis that patients with spliceosome mutations are more likely to respond to ATR inhibition and will explore potential genetic and epigenetic mechanisms of resistance. Future goals will be to identify rational combination strategies incorporating ATR inhibition in MDS treatment. Ongoing basic and translational research should provide mechanistic insights into R-loops and the ATR response, as well as additional targetable vulnerabilities and biomarkers that can be used for patient selection and to monitor responses. Furthermore, it will be important to address whether aberrant R-loop accumulation is common in other cancers harboring spliceosome mutations. If aberrant R-loop accumulation is a common vulnerability in cancers, ATR inhibitors may have a broad therapeutic potential in cancer therapy.


Transcription factors bind regulatory DNA sequences in a combinatorial manner to modulate gene expression. Deep neural networks (DNNs) can learn the cis-regulatory grammars encoded in regulatory DNA sequences associated with transcription factor binding and chromatin accessibility. Several feature attribution methods have been developed for estimating the predictive importance of individual features (nucleotides or motifs) in any input DNA sequence to its associated output prediction from a DNN model. However, these methods do not reveal higher-order feature interactions encoded by the models.in vivo TF binding models. DFIM reveals epistatic interactions involving nucleotides flanking the core motif of the Cbf1 TF in yeast from in vitro TF binding models. We also apply DFIM to regulatory sequence models of in vivo chromatin accessibility to reveal interactions between regulatory genetic variants and proximal motifs of target TFs as validated by TF binding quantitative trait loci. Our approach makes significant strides in improving the interpretability of deep learning models for genomics.We present a new method called Deep Feature Interaction Maps (DFIM) to efficiently estimate interactions between all pairs of features in any input DNA sequence. DFIM accurately identifies ground truth motif interactions embedded in simulated regulatory DNA sequences. DFIM identifies synergistic interactions between GATA1 and TAL1 motifs from https://github.com/kundajelab/dfim.Code is available at: Bioinformatics online. Genome-wide biochemical profiling experiments have revealed millions of putative regulatory elements in diverse cell states. These massive datasets have spurred the development of deep neural network (DNN) models to predict cell-type specific or context-specific molecular phenotypes such as TF binding, chromatin accessibility and gene expression from DNA sequence . Beyond Advances in feature attribution methods for DNNs have enabled the identification of predictive cis-regulatory patterns in DNA sequences used as input to the models. Feature attribution methods estimate the contribution (or importance) of features, such as individual nucleotides or contiguous subsequences (e.g. motifs), in an input DNA sequence to a model’s output prediction. A perturbation-based, forward-propagation approach known as in-silico mutagenesis (ISM) quantifies the importance of a nucleotide in an input DNA sequence as the maximal change in the output prediction from the DNN model when the observed nucleotide (e.g. a G) at that position is mutated to any of the alternative bases . ISM has been used to score the potential molecular impact of genetic variants in regulatory DNA sequences . HoweverCurrent feature attribution methods only provide the importance of individual features. They do not highlight predictive, higher-order feature interactions that are learned by the DNN model. Perturbation-based approaches such as ISM cannot scale to comprehensively score all pairwise and higher-order interactions between nucleotides or subsequence features. Recently, an efficient algorithm was proposed to calculate SHAP-based pairwise feature interaction scores  to estimate pairwise interactions between features (nucleotides or subsequences) in an input DNA sequence mapped to an associated regulatory phenotype by a neural network. We define a novel Feature Interaction Score (FIS) between any pair of features (source feature and target feature) in an input DNA sequence as the change in the importance score of the target feature when the source feature is perturbed, while keeping all the other features in the sequence intact. By leveraging efficient backpropagation-based feature attribution methods, we can efficiently compute FIS between all pairs of nucleotides or predictive motifs across large sets of input DNA sequence. Aggregate summary statistics of the pairwise Feature Interaction Score across multiple sequences provide insights into common, shared patterns of feature interactions.in vivo TF binding models. We apply DFIM to reveal epistatic interactions involving nucleotides flanking the core motif of the Cbf1 TF in yeast from in vitro TF binding models. We also apply DFIM to regulatory sequence models of in vivo chromatin accessibility to reveal interactions between regulatory genetic variants and proximal motifs of target TFs as validated by TF binding quantitative trait loci.We benchmark DFIM in controlled simulations that explicitly encode motif interactions. We use DFIM to reveal synergistic interactions between GATA1 and TAL1 motifs from X of length L to a categorical (binary or multiclass classification) or continuous (regression) output O. Let Y refer to the scalar predicted output O from the neural network for regression tasks. For classification tasks, let Y refer to the scalar input to the final output sigmoid (i.e. logit) of the neural network.We assume that we have trained a deep neural network to accurately map one-hot encoded DNA sequences We are given a one-hot encoded input DNA sequence on 1≤p≤L .X0, the output Y0 can be approximated by a first-order Taylor expansion w0 is the partial derivative (gradient) of Y with respect to the input sequence variable X evaluated at X0 i.e. w0 can be computed efficiently in one backpropagation pass. We then perform a point-wise multiplication of the gradient matrix w0 with the one-hot encoded observed input sequence X0 to obtain the importance scores for the observed nucleotides b at each position p i.e. X0 to those obtained when using neutral ‘reference’ sequences  and the observed nucleotide at some other position (target feature) in the sequence. Let X0 where we switch the observed nucleotide α at source position s to a different mutant nucleotide X0  of the target feature with the source feature as the maximal FIS marginalized over all possible values of the mutant nucleotide γ at the source feature We define the maximal Feature Interaction Score )  Step 5.maxFIS scores for all pairs of source and target features in an input DNA sequence (A nucleotide-resolution Deep Feature Interaction Map (DFIM) summarizes the sequence  Step 6.FIS between a source position s and target position t across a collection of input sequences iX, we first identify the subset of sequences FIS or absolute FIS corresponding to each S.  of all positions GCf i.e. the mutant sequence FIS of the target motif with the source motif is once again the difference of the sum of importance scores across all target nucleotides X0 and the mutated sequence To compute the FIS, across a collection of input sequences, we define statistically significant interactions based on an empirical null distribution of scores from dinucleotide shuffled versions of the input sequences. For each dinucleotide shuffled input sequence, we compute FIS for all nucleotide pairs. We fit a Gaussian distribution to this null empirical distribution of FIS scores. FIS values passing a P-value of 0.05 with respect to this null distribution are considered statistically significant. We use the Benjamini-Hochberg procedure for multiple hypothesis correction. Given a continuous distribution of F features (nucleotides/motifs), SHAP interaction scores scale at least quadratically to compute all pairwise interactions giving a complexity ranging from F source features, we need one more backpropagation pass to obtain FIS of that source with all target features. Thus, DFIM exhibits a complexity of O(F) scaling linearly in the number of features. Our proposed FIS is essentially an efficient approximation of SHAP interaction scores. Further, in contrast to SHAP interaction scores and pairwise ISM interaction scores which are necessarily symmetric over the source and target, FIS is directional and can produce asymmetric interaction scores.For an input sequence with FIS, we simulated 60 K random DNA sequences (0.46 G/C frequency) of length 200 bp. We divided these into 3 sets of 20 K sequences. We randomly embedded 1 or 2 instances of the ELF1 motif [using the highest affinity sequence from Position Weight Matrix  for all sequences in the positive class (i.e. Set 3). We used DeepLIFT with a fixed GC reference for computing importance scores since the underlying sequences were generated using a fixed GC background. Only pairs of SIX5 and ELF1 motifs (positive control) showed strong FIS (P < 0.05) P-values, even after multiple hypothesis correction. None of the other motif pairs show statistically significant interactions  in erythroid K562 cells from all other chromatin accessible DNase-seq peaks in K562 (negative class) (FIS (using DeepLIFT with shuffled reference as importance scores) for all pairs of GATA1, TAL1 motif instances across all sequences using GATA1 as the source motif. We observed several instances with strong FIS between proximal GATA1 and TAL1 motifs which corroborates their experimentally validated co-binding interactions  showed statistically significant higher interaction scores than all three other bins   . The CNNe class) , achievee class) . We thenractions A. To undarisons) A. Howevearisons) B, an obson GATA1 .FIS to investigate an orthogonal question—What proximal sequence features are affected by (interact with) regulatory genetic variants? Tehranchi et al. developed a pooling-based approach to identify thousands of SNVs that have allelic effects on TF binding (as measured by ChIP-seq) across a large collection of genotyped lymphoblastoid human cell-lines  CNN model to map 1kbp length DNA sequences to binary chromatin accessibility profiles across 16 primary hematopoietic cell types (with ATAC-seq data) . The modFIS to estimate the effect of mutating the reference allele to all alternate alleles at the source QTL on every target nucleotide ±15 bp around the QTL. First, we observed strong positive  but not the A allele  will destroy the importance of the entire JUND motif.To understand proximal interactions, for each bQTL, we used positive A and negpositive B interacpositive A. The reFIS predicts that the C allele (weaker binding) destroys the importance scores of the core GGAA element  and alternate allele C. The bQTL is in the flanking nucleotides of a low affinity SPI1 site where only the core GGAA matches the canonical motif.  element A. Tehran element A. E.g. a element B. SPI1 Q element .FIS of a matched number of conservative control SNVs from the FIS (maxAbsFIS) over all target nucleotides ±15 bp around the SNV. For all the TFs, we found that the bQTLs exhibit significantly (Mann Whitney test) stronger maxAbsFIS than control SNVs  and 5 variable flanking nucleotides on either side. They trained a feed forward neural network to predict relative binding affinity (ΔΔG) for each of the TFs from the 10 bp flanking sequences (using a flattened one-hot encoding) in the library (P = 0.25) with a final dense classification layer having a linear activation. They used a distillation approach to interpret the NN model by fitting a linear model with all mononucleotide features across all positions and all dinucleotide features across all pairs of positions to the output predictions of the NN. They found that dinucleotide features were critical for the linear model to have a good fit  that affect binding of transcription factors. Our maps link binding QTLs to nearby sequence features including high and low affinity matches to the canonical binding site of the TF whose binding is disrupted. We also find bQTLs interacting with motifs of multiple co-binding TFs. These epistatic interactions seem to capture both cooperation and competition. While our primary focus in this manuscript is on interpreting feature interactions in DNA sequence inputs, DFIM can easily be generalized to other data modalities.We present an efficient method called Deep Feature Interaction Maps (DFIM) to identify epistatic interactions between all pairs of nucleotides or motif features in any DNA sequence input to a deep learning model for regulatory genomics. Our method accurately recovers ground truth interacting motifs in simulated regulatory DNA sequences. When applied to deep learning models of Partial dependence plots are commonly used to understand the sensitivity of a prediction to a one or more features . DFIM seThere are some caveats to interpreting feature interactions derived from DFIM. Feature importance scores from any feature attribution method are only meaningful for examples that are predicted correctly. Since feature interaction scores from DFIM are based on feature importance scores, the validity of DFIM is also restricted to examples that are correctly predicted by high performance models. Further, vulnerabilities of the feature attribution method used in DFIM transfer over to the interaction scores. Hence, we recommend using multiple feature attribution methods to obtain robust estimates of interactions. Changes in model architecture can also change the interactions encoded by the model and thus the interactions learned with DFIM. Despite these mentioned caveats, the case studies we present here showcase the utility of DFIM to provide a nuanced view into the combinatorial code of regulatory DNA sequences through the lens of predictive neural network models.Supplementary DataClick here for additional data file.


A similar but less dramatic effect was observed with alpha-band (8–12 Hz) correlation changes . DISCUSSION/SIGNIFICANCE OF IMPACT: The negative correlation between inter-electrode power envelope correlations in the beta frequency band and motor recovery indicates that activity in the motor cortex on each hemisphere may become more independent during recovery. The role of the unaffected hemisphere in stroke recovery is currently under debate; there is conflicting evidence regarding whether it supports or inhibits the lesioned hemisphere. These findings may support the notion of interhemispheric inhibition, as we observe less in common between activity in the 2 hemispheres in patients successfully achieving recovery. Future neuroimaging studies with greater spatial resolution than available with EEG will shed further light on changes in interhemispheric communication that occur during stroke rehabilitation.OBJECTIVES/SPECIFIC AIMS: The objective of this study is to determine the degree to which the use of a contralesionally-controlled brain-computer interface for stroke rehabilitation drives change in interhemispheric motor cortical activity. METHODS/STUDY POPULATION: Ten chronic stroke patients were trained in the use of a brain-computer interface device for stroke recovery. Patients perform motor imagery to control the opening and closing of a motorized hand orthosis. This device was sent home with patients for 12 weeks, and patients were asked to use the device 1 hour per day, 5 days per week. The Action Research Arm Test (ARAT) was performed at 2-week intervals to assess motor function improvement. Before the active motor imagery task, patients were asked to quietly rest for 90 seconds before the task to calibrate recording equipment. EEG signals were acquired from 2 electrodes—one each centered over left and right primary motor cortex. Signals were preprocessed with a 60 Hz notch filter for environmental noise and referenced to the common average. Power envelopes for 1 Hz frequency bands (1–30 Hz) were calculated through Gabor wavelet convolution. Correlations between electrodes were then calculated for each frequency envelope on the first and last 5 runs, thus generating one correlation value per subject, per run. The chosen runs approximately correspond to the first and last week of device usage. These correlations were Fisher Z-transformed for comparison. The first and last 5 run correlations were averaged separately to estimate baseline and final correlation values. A difference was then calculated between these averages to determine correlation change for each frequency. The relationship between beta-band correlation changes (13–30 Hz) and the change in ARAT score was determined by calculating a Pearson correlation. RESULTS/ANTICIPATED RESULTS: Beta-band inter-electrode correlations tended to decrease more in patients achieving greater motor recovery (Pearson’s


Individuals who received anterior cruciate ligament (ACL) reconstruction surgeries demonstrated lower extremity movement asymmetries. The purpose of this study was to determine if psychological impairment was a contributor to lower extremity movement asymmetries in walking for individuals who received ACL reconstruction surgeries. Three-dimensional videographic and force plate data were collected for 25 males after unilateral ACL reconstruction performing walking without (single-task condition) and with the concurrent cognitive task . Both uninjured and injured legs had significantly smaller peak knee flexion angle and peak knee extension moment during loading response and mid-stance phases in dual-task condition compared to single task condition (P ≤ 0.012). Walking condition and leg had significant interaction effects on peak hip adduction angle during mid-stance phase (P = 0.042) and peak hip abduction moment during loading response phase (P = 0.048). The inter-leg difference of peak hip adduction angle during mid-stance (P = 0.038) and terminal stance (P = 0.036) phases, and peak hip abduction moment during loading response phase (P = 0.024) were significantly decreased in dual-task condition compared to single-task condition. Psychological factors have significant effects on post-operative movements of both injured and uninjured knees of individuals who received ACL reconstruction surgery. Although physical factors may be primary contributors to the post-operative lower extremity movement asymmetries, psychological factors also contribute to the post-operative hip movement asymmetries. Approximately 175000 ACL reconstruction surgeries were performed in the United States annually2. Although ACL reconstruction surgeries restored knee stability, studies repeatedly reported high ACL re-injury rates and early development of knee osteoarthritis (OA) after ACL reconstruction. Literatures showed that up 12% of patients with reconstructed ACLs had ACL re-injury4 and the prevalence of radiographic knee OA of patients with combined ACL and meniscal injuries in 10 to 15 years follow-up after ACL reconstruction were 80%5.Anterior cruciate ligament (ACL) reconstruction is a commonly surgical procedure to restore knee functions after an ACL injury8. Literature showed that athletes with multiplane biomechanical asymmetries at hip and knee at the time of return to sport were at least three times more likely to have ACL re-injury within one year compared to those without these asymmetries7. Literature also showed that asymmetrical lower extremity loading altered chondrocyte synthesis and catabolic activities, which could lead to structural damage of articular cartilage and may accelerate the development of knee OA11.Lower extremity movement asymmetries were found to be predictive of ACL re-injury and knee OA after ACL reconstruction13. Patients with quadriceps strength asymmetry demonstrated increased asymmetry in lower extremity kinetics in functional activities compared to patients with quadriceps strength symmetry15. These studies appear to indicate impaired quadriceps strength was the contributor to lower extremity movement asymmetries after ACL reconstruction. A study by Roewer et al., however, showed that asymmetric knee angles, knee moments, and hip power still existed despite subjects achieving symmetric quadriceps strength after ACL reconstruction16. Lentz et al. also reported that despite quadriceps strength was improved from 6 months to 1 year, functional limitations remained unchanged17. These studies indicated that quadriceps strength deficit was not the only contributor to lower extremity movement asymmetries.Weak quadriceps strength was identified as a primary contributor to lower extremity movement asymmetries after ACL reconstruction surgery. Studies showed that impaired quadriceps strength was correlated to altered movement patterns of injured leg after ACL reconstruction surgery18. A meta-analysis of the state of play revealed that fear for re-injury was the most common reason for post-operative reduction in or cessation of sports participation19. A study on psychological changes of patients who received ACL reconstruction surgery found that patients who returned to the competitive sport at 12 months after the surgery had more positive psychological responses to sports participation at 6 and 12 months after the surgery compared to patients who had not returned to competitive sport20. Although these studies indicated that psychological impairments may contribute to patients’ movement asymmetries after ACL reconstruction surgery, the effect of psychological impairment on lower extremity movement asymmetries has not been confirmed yet.In addition to physical impairments, psychological impairments could also contribute to lower extremity movement asymmetries after ACL reconstruction. Movement asymmetries after ACL reconstruction surgery may be due to patients’ preference of using their uninjured leg, which could put the contralateral leg at high risk for injuryThe purpose of this study was to determine if psychological impairment was a contributor to lower extremity movement asymmetries in walking for individuals who received ACL reconstruction surgeries. We hypothesized that there would be a quadriceps strength asymmetry after ACL reconstruction surgery. We also hypothesized that there would be asymmetries in hip and knee angles and moments in walking after ACL reconstruction surgery. We further hypothesized that asymmetries of hip and knee angles and moments would be decreased when walking with a cognitive task compared to walking without the cognitive task.Twenty-five males who received unilateral ACL reconstruction using hamstring tendon autograft within 10 months before the study  volunteered to participate in this study. All participants had isolated ACL injury without other ligament and meniscus injuries that needed to be repaired with a resection or suture, or moderate or severe articular cartilage damage to the patellofemoral and tibiofemoral joint, or other current orthopaedic injuries or disorders that were still affecting lower extremity movements. All participants went through similar post-operative home-based rehabilitation programs and had normal knee range of motion at the time of participating in the study. The mean International Knee Documentation Committee (IKDC) score of the participants was 67.4 ± 11.4. The use of human subjects in this study was approved by the Institutional Review Board of Peking University Third Hospital. All participants read and signed a written informed consent before data collection.Each participant was asked to wear a pair of black spandex shorts. Passive reflective markers were placed bilaterally at the anterior superior iliac spine, posterior superior iliac spine, lateral thigh, lateral femoral condyle, medial femoral condyle, anterior superior shank, anterior inferior shank, lateral malleoli, medial malleoli, heel, and first and fifth metatarsophalangeal. The participant was instructed to walk with barefoot in two conditions: (1) without cognitive task (single-task condition), and (2) with concurrent cognitive task . In single-task condition, the participant walked along a 10-m walkway at self-selected walking speed. In dual-task condition, the participant walked along the walkway while backward counting numbers with an increment of seven starting from a randomly given number between 125 and 250. In dual-task condition, participants immediately initiated the walking task at their preferred speed once they heard the number read by the investigator and kept counting out loudly as required without repeating the given number. Participants were asked to count as fast as possible while walking task. A 5 min rest was requested between single-task and dual-task test. The order of conditions was randomized for the participant. Quadriceps strength was evaluated for the participant after walking test.Three-dimensional (3-D) trajectories of the reflective markers were collected using an 8-camera motion capture system  at a sample rate of 100 Hz. Ground-reaction force signals were collected using two embedded force plates  at a sample rate of 1000 Hz. Each participant was asked to have three successful trials for each condition. A successful trial was defined as a trial in which the participant performed the task as required, and all kinematic and kinetic data were collected.Quadriceps isometric strength was quantified with an isokinetic dynamometer  during a maximum voluntary isometric contraction (MVIC). The participant was seated with a hip flexed at 90° and knee flexed at 60°. The lateral femoral condyle was aligned with the dynamometer axis, and the dynamometer resistance pad was secured to the anterior aspect of the distal shank. After correcting for leg weight, the participant was asked to perform submaximal practice to familiarize themselves with the testing apparatus. After familiarization of testing apparatus, the participant was asked to have three recorded maximum-effort trials  for each leg with uninjured leg tested first.21. Kinematics and kinetics variables were calculated using Visual 3D software . Hip and knee joint angles were calculated as Cardan angles of distal segment reference frame relative to the proximal segment reference frame in an order of (1) flexion-extension, (2) abduction-adduction, and (3) internal-external rotation. Joint moments were calculated through an inverse dynamic approach and transferred into the distal segment reference frame. Joint moments and quadriceps strength were normalized to the production of body weight and standing height.The raw 3-D trajectories of the reflective markers were filtered through a Butterworth low-pass digital filter at an estimated optional cut-off frequency of 10 HzLstride length was the forward distance of heel marker between successive heel contacts of the same foot, Tcycle time was the time between the first contacts of 2 consecutive footfalls of the same foot.Three phases, loading response phase, mid-stance phase, and terminal stance phase, during walking were analyzed Fig. . Knee ra22Yuninjured leg and Yinjured leg were magnitudes of a given joint angle or moment of uninjured leg and injured leg.The inter-leg difference (ILD) between injured and uninjured legs was also calculated for joint angles and moments as described in literaturePaired t-test was performed to compare the walking speed between single-task and dual-task conditions and isometric quadriceps strength between injured and uninjured legs. Two-way analyses of variance (ANOVA) with repeated measures were performed to compare the kinematic and kinetic variables of interest between conditions  and legs (injured leg and uninjured leg). In case a significant interaction effect of condition and leg was detected, paired t-tests were performed to compare the dependent variables between legs in a given condition and between conditions on a given leg. Paired t-tests were performed to compare the ILD between single-task and dual-task conditions. All statistical analyses were performed using SPSS computer program package, version 16.0 . A type I error rate less than or equal to 0.05 was considered as the indication of statistical significance.Walking speed in dual task condition (1.15 ± 0.13 m/s) was significantly lower than that in single task condition (1.24 ± 0.14 m/s) (P = 0.003). The quadriceps strength of injured leg (0.096 ± 0.028 BW × BH) was significantly smaller than that of uninjured leg (0.129 ± 0.028 BW × BH) (P = 0.001).No significant interaction effect of condition and leg on knee angular kinematics were detected P ≥ 0.162). Injured leg had smaller peak knee flexion angle during loading response and mid-stance phases compared to uninjured leg  (Table 62. InjurNo significant interaction effects of condition and leg on knee moments were detected (P ≥ 0.193). Injured leg had smaller peak knee flexion moment during terminal stance phase compared to uninjured leg (P = 0.001) Table . Both unA significant interaction effect of condition and leg on peak hip adduction angle during mid-stance phase was detected (P = 0.042). Injured leg had significant smaller peak hip adduction angle during mid-stance phase compared to uninjured leg in single-task condition (P = 0.050) while no significant difference was found in this variable between legs in dual-task condition (P = 0.256) Table . Both unNo significant interaction effects of condition and leg on other hip angular kinematics were detected (P ≥ 0.055). Injured leg had smaller minimum hip flexion angle during loading response phase compared to uninjured leg (P = 0.004) Table . InjuredA significant interaction effect of condition and leg on peak hip abduction moment during loading response phase was detected P = 0.048). Injured leg had significantly smaller peak hip abduction moment during loading response phase compared to uninjured leg in both single task and dual task condition  (Table 8. InjureNo significant interaction effects of condition and leg on other hip moments were detected P ≥ 0.214). Injured leg had smaller peak hip abduction moment during mid-stance phase compared to uninjured leg (P = 0.023) (Table . Injured24. These results of this study combined with the literatures indicated that participants in this study indeed had quadriceps asymmetries as defined in literatures. A previous study demonstrated that increased quadriceps strength was associated with increased knee cartilage and greater cartilage cross-sectional area25, indicating quadriceps weakness as a possible contributor to the development of knee osteoarthritis of individuals received ACL reconstruction surgeries. Extended rehabilitation programs, therefore, are needed to correct quadriceps strength asymmetry after ACL reconstruction surgery to minimize the risk of early knee osteoarthritis.The results of this study support our hypothesis that there would be a quadriceps strength asymmetry after ACL reconstruction surgery. The decreased quadriceps strength of injured leg in this study was consistent with the quadriceps strength asymmetries reported in literatures28. These results suggested that individuals who received ACL reconstruction surgeries tended to walk with increased leg stiffness29. The results of this study further showed that injured leg had smaller peak knee flexion moment during terminal stance phase while having no significant difference in peak knee abduction moments during mid-stance and terminal stance phases between injured and uninjured legs, which is also consisitent with literature31. Literatures show that knee moment in the sagittal plane is positively correlated with medial femoral cartilage thickness in healthy individuals32, and that approximately 85% of the mechanical work during gait is done in the sagittal plane33. The sagittal-plane moments appears to be more revelant to the knee joint degeneration after ACL reconstruction surgery than peak knee abduction moment. The results of current study also showed that injured leg had smaller peak hip abduction moment during the mid-stance phase compared to uninjured leg while having no significant difference in peak hip abduction moment during terminal stance phase. The decreased peak hip abduction moment in the mid-stance phase in this study was likely due to weaker hip abductors. Hip abductors assist in stabilizing pelvis in the frontal plane. Ipsilateral hip abductor weakness may cause the pelvis to drop towards the contralateral swing leg34. A recent study showed that patients who develop medial compartment knee OA within 5 years after ACL reconstruction surgery walked with asymmetrically lower peak hip abduction moments compared to those who did not develop knee OA at 5 years35. These results combined together suggested that altered hip movement patterns in the frontal plane may be a vital predictor to initiate knee degeneration and may need to be addressed during rehabilitation.The results of this study support our hypothesis that there would be asymmetries in hip and knee motion patterns in walking after ACL reconstruction surgery. The decreased peak knee flexion angle during the loading response phase of injured leg observed in this study is consistent with literatureset al. reported that patients after ACL reconstruction surgery increased brain activation in attentional and sensory areas during force reproduction task compared to the healthy control group, which reflect an increased focused attention and therefore an increased neurocognitive involvement36. According to the limited capacity theory of attention, when an attention-demanding cognitive task imposed on an attention-demanding task, concurrent execution of both tasks may require more attentional capacity than the existing capacity and therefore one or both tasks could not have sufficient attention37. Walking while performing a cognitive task decreased attention distribution to walking in the dual task condition compared to single task condition in this study. In this situation, the decreased ILD should be considered as a result of decreased psychological effects on asymmetries of walking. These results, therefore, indicate that psychological as well as physical factors contribute to movement asymmetries post ACL reconstruction surgeries.The results of this study partially support our hypothesis that asymmetries of hip and knee angular kinematics and moments would be decreased when walking with a cognitive task compared to walking without the cognitive task. Interaction effects of condition and leg on hip adduction angles and abduction moments and decreased corresponding ILDs observed this study suggested that asymmetries of hip movements in the frontal plane was decreased while walking in dual task condition. No similar results were obtained for knee angles and moments. Baumeister 38. Considering these results, although we don’t know the cost of the performance of the cognitive task when participants walking in dual task condition in current study, the results of current study indicated that performing cognitive task while walking was indeed at the cost of the performance of gait for individuals who received ACL reconstruction surgery, which is similar to that of patients with Parkinson’s disease, but different from that of healthy individuals. This means that psychological factors still affect post-operative gait for individuals who received ACL reconstruction surgeries, but this effect may be similar for both injured and uninjured legs.Although the asymmetries of knee angular kinematics and moments were not decreased while walking with a cognitive task, post-operative knee movements of patients who received ACL reconstruction surgery were still affected by psychological factors. The results of this study showed that participants significantly decreased their knee angles and moments while walking with a cognitive task. A study showed that healthy individuals prioritized gait control at the cost of performance of the cognitive task in dual task condition, while patients with Parkinson’s disease prioritized performance of the cognitive task at the cost of performance of gaitet al. suggested that postural stability in the frontal plane was controlled primarily by activation of the hip abductors and adductors designed to alternately load and unload the hips, thereby controlling the movement of center of mass39. Abnormal hip movements could have a deleterious effect on the tibiofemoral and patellofemoral mechanics in multiple planes40. This means that the effects of psychological factors on lower extremity movements could be mainly through their effects on the movements of hip joints. Interventions which address proximal movement impairments may be beneficial for patients who had various knee movement impairments. As participants had lower quadriceps strength of injured leg compared to uninjured leg in this study, their knee movements might be mainly affected by physical factors while hip movements affected by both physical as well as psychological factors.Psychological factors might have different influence on post-operative hip and knee movement patterns in walking for individuals who received ACL reconstruction surgeries. The results of this study showed that ILDs of hip angular kinematics and moments in the frontal plane were decreased in dual task condition compared to single task condition, while ILDs of knee angular kinematics and moments were essentially the same between conditions. These results indicated that effects of psychological factors on asymmetries of hip and knee movements are different. The hip joint is the most proximal joint in the lower extremity kinematic chain and shares the femur with the knee joint. Winter 41, likely due to either poor movement strategies or lingering impairments that were not addressed by surgical reconstruction and post-surgery rehabilitation. Movement strategies are modifiable and represent important targets for successful clinical interventions. The results of this study indicated that psychological factors are contributors to the post-operative lower extremity movement asymmetries of individuals who received ACL reconstruction surgeries. As psychological differences regarding returning to sports among athletes who received surgeries treating injuries occur as early as six months postoperatively20. Clinicians should address psychological factors early to help patients modify the abnormal movement patterns and facilitate their returns to competitive sport. Though the results of this study indicated that the immediate effect of the secondary cognitive task did not decrease the asymmetry of knee movements, the asymmetry of hip movements in frontal plane could be modified in dual-task condition. Dual-task training could be used as an intervention to improve movement performance and motor control in rehabilitation programs42.Our findings have important implications for post ACL reconstruction rehabilitation programs. The risk of ACL re-injury remains higher than that of the initial ACL injury15, the effect of the concurrent cognitive task on walking may be different between the group with symmetrical quadriceps strength and the group with asymmetrical quadriceps strength. Future studies may be needed to assess the effects of physical factors on the effects psychological factors on lower extremity movement asymmetries. In addition, since gender differences in lower extremity movement patterns exist43, psychological factors may affect lower extremity movement asymmetries differently between genders. Future studies should include both females and males to investigate the gender differences in psychology.Several limitations of this study may need to be addressed in the future studies. Since the asymmetry of quadriceps strength could affect the lower extremity movement symmetryThere were significant post-operative asymmetries in quadriceps strength, and hip and knee movements of individuals who received ACL reconstruction surgery.Psychological factors have significant effects on post-operative movements of both injured and uninjured knees of individuals who received ACL reconstruction surgery.Although physical factors may be primary contributors to the post-operative lower extremity movement asymmetries, psychological factors also contribute to the post-operative hip movement asymmetries.The results of this study appear to warrant the following conclusions:


The high-order structure of metaphase chromosomes remains still under investigation, especially the 30-nm structure that is still controversial. Advanced 3D imaging has provided useful information for our understanding of this detailed structure. It is evident that new technologies together with improved sample preparations and image analyses should be adequately combined. This mini review highlights 3D imaging used for chromosome analysis so far with future imaging directions also highlighted. The DNA in chromosomes holds the genomic information of all eukaryotes. Throughout the cell cycle progression, chromosomes condense into the higher-order compact structures until they reach mitosis. The biological implications of this chromosome higher-order structure is for packaging the long DNA molecules into chromatin fibers within chromosomes facilitating upon cell division, the separation of the chromatids for transportation of DNA fibers evenly to two new daughter cells without damage Sumner . The higThe 30-nm chromatin fibers have frequently been observed by scanning electron microscopy (SEM)  has Angstrom resolution, it is not a suitable microscopy for studying intact mitotic chromosomes because chromosomes are too thick for the electrons to penetrate through biological samples over 1 micron thick Ris . Whole hScanning electron microscopy (SEM) images the sample surface by scanning it with a focused electron beam and collection of backscattered electrons. The major limitation has been due to sample preparation as MAA chromosomes show an artificial surface “skin” layer over the chromosomes following air-drying   allows 3D information and is similar to FIB-SEM but uses a diamond knife instead of focused GIn contrast to TEM, X-rays have the ability to penetrate through whole intact chromosomes. Imaging intact chromosomes is advantageous and simplifies the procedure saving time for sample preparation. Coherent diffraction imaging (CDI) using X-ray diffraction was achieved on intact mitotic chromosomes in both 2D and 3D  may be another strong tool to visualize 3D chromosome structure. AFM gives us images which are similar to those obtained using SEM by scanning the solid sample surface with a sharp probe tip and detecting the interaction force between the tip and the sample to get height information. From this principle, AFM does not require metal coating and the observation can be performed either in vacuum, air, or liquid condition , a super-resolution technique, has been performed on mitotic chromosomes Carlton . SIM allDespite the impressive nature of imaging methods mentioned above, the detailed higher-order structure is still under debate. Optical super-resolution microscopy techniques are under development and present an opportunity to study the structure of the chromosomes below the diffraction limit. Such methods should allow the study of live cells that has not been possible with any previous high-resolution method and with development in 3D. Optical super-resolution techniques are already being applied to study chromatin contacts  after labelling with H2AvD-EGFP, a histone H2A variant in the presence of background (Matsuda et al. 3D studies done to date have tended to require harsh chemical fixation, dehydration, and drying steps that may not represent the true nature of the sample under investigation. Cryo-electron microscopy enables us to observe biological samples close to the native state (Eltsov et al.


Linkage disequilibrium (LD)—the non-random association of alleles at different loci—defines population-specific haplotypes which vary by genomic ancestry. Assessment of allelic frequencies and LD patterns from a variety of ancestral populations enables researchers to better understand population histories as well as improve genetic understanding of diseases in which risk varies by ethnicity.We created an interactive web module which allows for quick geographic visualization of linkage disequilibrium (LD) patterns between two user-specified germline variants across geographic populations included in the 1000 Genomes Project. Interactive maps and a downloadable, sortable summary table allow researchers to easily compute and compare allele frequencies and LD statistics of dbSNP catalogued variants. The geographic mapping of each SNP’s allele frequencies by population as well as visualization of LD statistics allows the user to easily trace geographic allelic correlation patterns and examine population-specific differences.https://ldlink.nci.nih.gov/?tab=ldpopLDpop is a free and publicly available cross-platform web tool which can be accessed online at  Linkage disequilibrium (LD)—the non-random association of alleles at different loci—defines population-specific haplotypes which vary by genomic ancestry . Assessm2) for each included population. The reference genetic data is from the 1000 Genomes Project Phase 3, which includes sequencing data for 2504 individuals in 26 ancestral populations which are divided into 5 super populations  [ftp://ftp.1000genomes.ebi.ac.uk/vol1/ftp/release/20130502/).LDpop is an online module designed to allow researchers to query LD patterns of two variants across ancestral populations of interest. LDpop estimates allele frequencies and measures of LD D′ and R for eachhttps://github.com/CBIIT/nci-webtools-dceg-linkage/.LDpop is written in Python 2.7.15) and runs on a web-accessible virtual machine with UNIX operating system. The genomic coordinates are retrieved for each query variant from an indexed MongoDb database of dbSNP version 151 and subsequently extracted from the phased 1000 Genomes Project variant call format (VCF) file using Tabix (0.2.5). LDpop uses the Google Maps API to produce the interactive geographic mapping for each population using latitude and longitudinal coordinates for each 1000 Genomes Project ancestral population. The LDpop web-accessible page is programed in HTML5 for cross-browser and cross-platform compatibility and is part of the larger LDlink collection of LD web tools  and runs. All cod2) to report for the geographic mapping. LDpop supports queried dbSNP variants which are biallelic including both single nucleotide polymorphism (SNP) and small insertion and deletion (indel) queries. LDpop allows the user to specify any subset of populations from the subpopulations, super populations, and all populations, they are interested in examining for the analysis.LDpop takes as input two dbSNP reference SNP numbers (rsIDs), a selection of desired populations from the 1000 Genomes Project, and a choice of which LD statistic (D′ or R2) for the two query variants for every population selected. The mapped data point is colored in proportion to the gradient shown in the legend, with darker red signifying a higher degree of LD. All geographic mapping utilizes the Google Maps API for smooth and rapid performance. The interactive summary table at the bottom of the page has a row for each selected 1000 Genomes Project population and displays data in the number of samples in each population, allele frequencies for each variant, and calculated LD values (D′ and R2). This table is sortable by column and has a search bar to quickly navigate through it. The table is also downloadable as a text file for local storage and future data integration and analysis.LDpop produces three geographic maps and one sortable, filterable table as output Fig. . For eachttps://ldlink.nci.nih.gov/?tab=ldpop.LDpop is an online module designed to allow researchers to query LD patterns of two variants across ancestral populations of interest. It is designed to allow users to easily calculate and geographically visualize these LD patterns and changes in allele frequency across ancestral populations. This web tool is freely available and can be accessed at


Nitrogen (N) loss from agricultural systems raises concerns about the potential impact of farming practices on environmental quality. N is a critical input to agricultural production. However, there is little understanding of the interactions among crop water use, N application rates, and soil types. This study was designed to quantify these interactions in corn (Zea mays L.) grown in production-size fields in central Iowa on the Clarion-Nicollet-Webster soil association. Seasonal water use varied by soil type and N application rate. Yield varied with N application rate, with the highest average yield obtained at 100 kg ha. N use efficiency (NUE) decreased with increasing N application rates, having values around 50%. Water use efficiency (WUE) decreased as N fertilizer rates increased. Analysis of plant growth patterns showed that in the low organic matter soils (lower water-holding capacities), potential yield was not achieved because of water deficits during the grain-filling period. Using precipitation data coupled with daily water use throughout the season, lower organic matter soils showed these soils began to drain earlier in the spring and continued to drain more water throughout the season. The low NUE in these soils together with increased drainage lead to greater N loss from these soils. Improved management decisions have shown that it is possible to couple water use patterns with N application to increase both WUE and NUE.


Hospitalization of older adults with cognitive impairment (CI) has been associated with higher risk for adverse outcomes. Acute Care for Elders (ACE) Units were developed to meet the unique hospital care needs of older adults and have been associated with reductions in functional decline and readmissions. The Virtual ACE intervention was developed to disseminate ACE principles across hospital units. Virtual ACE included training interprofessional providers to utilize screens and care protocols to optimize care for older adults on eight units at a large academic medical center. We conducted a preliminary analysis of mobility and patient outcomes before and after Virtual ACE among 192 older adults with CI on hospital admission. Chi-Square tests were used to examine the associations between Virtual ACE and patient outcomes. There were statistically significant pre vs. post improvements in patients’ mobility from bed to chair  and on the unit hallway . Although not statistically significant, there were also improvements in hospital room mobility  and documentation of activities of daily living (ADL) screens . There were non-significant reductions in pressure ulcer prevalence (26% vs. 22%) and restraint use (5% vs. 0%) during the hospital stay. Pain was similar before and after Virtual ACE. Virtual ACE was associated with increased mobility and slight reductions in adverse outcomes. As increased hospital mobility improves patient functioning post-discharge, Virtual ACE has the potential to maintain function and enhance outcomes in hospitalized older adults with CI.


OBJECTIVES/SPECIFIC AIMS: The goal of this study is to evaluate a potential strategy to overcome tamoxifen (tam) resistance by using tam in combination with high-intensity focused ultrasound (HIFU). Tam is the most commonly used anti-cancer therapeutic agent in estrogen receptor positive (ER+) breast cancer (BC) which accounts for ~70% of BC cases. Tam treatment decreases a woman’s risk of recurrence by 50%; however, BC that is initially responsive to tam often develops resistance. METHODS/STUDY POPULATION: HIFU deposits acoustic energy locally to a cancerous region, which induces strong vibrations of molecules inside and outside of the cells. The resulting absorption causes rapid heating and mechanical disruption. This clinically relevant, noninvasive, and nonionizing physical force modality, has been shown to synergistically enhance chemical anticancer therapies. RESULTS/ANTICIPATED RESULTS: In this study we found that treatment of MCF7 cells with HIFU and tam has additive antiproliferative effects and mediates increased cell death. Additionally, we used tam resistant (TR) MCF7 cells that had been exposed to low-dose tam over time until they acquired resistance. When MCF7 TR are treated with tam there is no change in viability; however, treatment with HIFU in combination with tam decreased viability of both MCF7 and MCF7 TR to 19% and the viability of the cell lines was indistinguishable. We next evaluated the effect on MCF7 Y537S mutant ESR1, where ER is mutated to be constitutively active. Treatment of MCF7 Y537S had no significant decrease in viability of combination therapy compared with viability after HIFU alone. Analysis of ERalpha gene expression showed that HIFU treatment increased ERalpha expression in MCF7 TR cells, thus resensitizing these cells to tam and allowing these therapies to work synergistically. Our team developed a system to evaluate the potential of this combination of therapies in a patient-derived xenografts (PDX) model. PDX have emerged as a novel translational tool for cancer research with the potential to more accurately recapitulate the molecular and behavioral aspects of cancer. The WHIM20 PDX is a tamoxifen resistant tumor where the patient developed the Y537S mutation in ESR1. Ex vivo experiments on PDX tumor pieces demonstrated that combination therapy of HIFU and tam work synergistically to increase cell death of these tumors. Further, cryogenic-scanning electron microscopy was utilized to directly demonstrate the physical disruption to both cellular and tumor microenvironment post exposure to combination treatment. DISCUSSION/SIGNIFICANCE OF IMPACT: These studies present a novel translational strategy to overcome tamoxifen resistance in ER+BC.


A 13-year-old child had left orbital trauma by wood. He consulted 2 week after for decreased visual acuity and eye pain. The ophthalmological examination showed the presence of a wound of autotight cornea a wooden foreign body incarcerated in the iris and the lens. Extraction of the foreign body associated with an adapted antibiotic treatment led to clinical improvement.


Killian-Jamieson diverticulum (KJD) is a rare entity resulting from the mucosal herniation through a lateral area of weakness known as the Killian-Jamieson space below the cricopharyngeal muscle. Ultrasonographic diagnosis is exceptional. Moreover, symptoms are found in only eleven percent of patients. KJD in its symptomatic form must be correctly identified because its therapeutic management is in some points different of Zenker’s diverticulum. A 73-year-old woman was suffering of mild dysphagia with occasional regurgitations and depressible swelling during swallowing on the left side of her neck which appeared two years earlier. Ultrasonographic examination showed a hypoechoic mass surrounded by a digestive wall measuring 45mm located just behind the left thyroid lobe Fig. . No concKJD is rare entity related to the more common Zenker’s diverticulum (ZD) with its classic posterior development. It results from the mucosal herniation through a lateral area of weakness known as the Killian-Jamieson space below the cricopharyngeal muscle . This spThe authors declare that they have no competing interests.


The characteristics of where older adults live have strong links with disability. Although lower income older adults experience disability at higher rates, less is known about the link between housing characteristics and functional health in this group. A within group comparison among this population is needed to understand how aspects of this vulnerable subgroup’s housing context are associated with health outcomes. The present study examines the association between housing and functional health among a U.S. nationally representative sample of independent living  lower income adults age 65+. Using data from round one of the National Health and Aging Trends Study, a sub-sample of N=2,865 lower income  older adults was selected for analysis. Regression analyses indicate that lower income older adults living in multiunit buildings reported better functional health compared to those in other housing contexts . This link also significantly varied by age and gender. Living in multiunit housing was associated with better functional health among those age 90+, not associated among those age 80-90, and was negatively associated among those age 65-79. In terms of gender, the link between multiunit housing and better functional health was only significant among women. Findings highlight variation in health across lower income older adults’ housing contexts. Potential explanatory mechanisms  will be discussed. Such information can inform senior housing policy regarding best approaches to providing housing for older adults that optimizes and promotes independent living.


Women diagnosed with Triple Negative Breast Cancer (TNBC) have worse overall survival rates than other forms of breast cancer partly due to the absence of novel and targeted treatment modalities in this aggressive subtype . The intAs an influential oncogene, MYC can potentially regulate ∼over 4,000 genes at any given moment. Although the relationship between MYC and DNA repair has been investigated extensively, therapeutic modalities that exploit MYC directed DNA repair vulnerability are absent. Our study cultivates the direct link between MYC and DNA repair to exploit an increased response to PARP inhibitors . EvaluatIn vitro and in vivo analysis of MYC downregulation via genomic and therapeutic inhibition induced PARP inhibitor sensitivity independent of BRCA status. The pan CDK  inhibitor dinaciclib demonstrated superior efficacy versus other MYC inhibitors in combination with PARP inhibition. The use of MYC as a driver of biomarker targeted therapy for TNBC provides hope for a patient population lacking in therapeutic treatment options while also addressing a validated association between MYC and TNBC, with approximately ∼50% of TNBC patients demonstrating upregulation of MYC in tumors [After reinforcing the relationship between MYC and the DNA repair pathway via RAD51, our study demonstrated a regulatory relationship between MYC and RAD51 gene expression . In vitrn tumors . Our anan CDK 1, , 5, 9 inThe relationship between CDK inhibition and MYC expression has illuminated a regulatory role of the cell cycle over MYC expression Figure . AdditioOur study also alludes to the evolution of PARP inhibitors in the clinical setting beyond BRCA status. Respected for their efficacious treatment responses in HR defect tumors with measurably decreased adverse events versus standard chemotherapy, PARP inhibitors offer a new horizon for cancer treatment . The shiMore recently, Sun et al. demonstrated that BRD4 inhibition induces PARP inhibitor sensitivity in ovarian cancers independent of BRCA status . Our stuin vivo. The down regulation of PD-L1 signals that checkpoint inhibitors may work in combination with dual dinaciclib + PARP inhibition. Two recent studies support the role that MYC plays in immune regulation; specifically MYC cooperates with KRAS to drive tumor growth via IL-23 and CCL9 immune suppression [To further advance the clinical application of PARP inhibitors, the investigations into novel immune-oncology agents are promising Figure . Studiespression . In anotpression . These sDinaciclib is a pan-CDK inhibitor that has not made it past Phase 2 clinical trial evaluation due to high toxicity . The reaAs we search for novel combination therapies that increase the efficacy and scope of the patient population eligible for PARP inhibitor treatment, it is paramount to remain focused on a clearly defined outcome. The use of efficacious biomarkers such as MYC that dictate response to therapy is essential to the evolution of targeted treatment option for TNBC patients Figure . We unde


A 50-year-old woman with widely metastatic melanoma presented to the emergency department with dyspnea. She was found to be tachypneic, hypoxic, tachycardic, and hypotensive. A non-rebreather oxygen mask was placed and her oxygen saturation improved mildly. We obtained a semi-erect chest radiograph (CXR) followed by chest computed tomography angiography (CTA) –3, due tThe diagnosis was a tension hydrothorax due to a massive, malignant pleural effusion causing hemodynamic compromise. The hemodynamic effects of a tension hydrothorax are analogous to those of a tension pneumothorax. Elevated intrathoracic pressure caused by the effusion impairs venous return and compresses the left ventricle, causing reduced stroke volume and subsequent hypotension.6A CXR will show a large pleural effusion.The emergency physician must be aware of this entity when a hemodynamically-compromised patient arrives with a pleural effusion, even if mediastinal shift is not evident on the CXR.What do we already know about this clinical entity?A tension hydrothorax due to a massive pleural effusion is a rare event that can cause hemodynamic instability due to compromised thoracic venous return.What is the major impact of the image(s)?The images demonstrate how increased intrathoracic pressure can cause mediastinal shift and vena cava compression.How might this improve emergency medicine practice?In a hemodynamically compromised patient with a large pleural effusion, emergent thoracentesis should be considered due to possible tension physiology.