Abstract Candida auris is a multidrug resistant pathogen that causes healthcare associated invasive infections and outbreaks with high mortality rates, often involving intensive care units (ICU). High rates of C. auris colonization among patients in ICUs are associated with increased risk of subsequent C. auris hospital-acquired infections. In this study, microbiological characteristics, and antifungal susceptibility patterns of C. auris isolates collected between January and December 2023 were investigated. Whole-genome sequencing was used to examine the genetic variation and molecular epidemiological characteristics. During the study period, a total of 20 patients were found to be colonized with C. auris by three site skin swab and eight of these patients subsequently developed invasive infection with C. auris. Among patients with both colonization and infection, all invasive isolates were closely related genetically to colonizing isolates and belonged to clade I. Candidemia was the most common infection identified in 4/8 (50%) patients. All isolates were resistant to fluconazole and thirteen isolates from six patients were resistant to amphotericin B. Paired isolates from patients with both colonization and invasive infection showed the same antifungal resistance patterns. All isolates had the ERG11 p.K143R mutation known to cause fluconazole resistance. Three isolates from one patient with a FKS1 hotspot region 3 p.M690I mutation were resistant to caspofungin and two isolates from one patient with a FKS1 hotspot region 2 p.R1354H mutation were resistant to caspofungin, anidulafungin and micafungin. Our study demonstrates that colonization with C. auris is associated with increased risk of infection by the same strain amongst critical ill patients. C. auris screening and implementation of adequate infection control measures are urgently needed to prevent the dissemination of C. auris infections.

PurposeKnee joint distraction is a surgical procedure with cartilage-regenerating properties. The composition of joint distraction-regenerated cartilage in human patients is poorly documented. In this case-study, provided a unique opportunity to biomolecularly characterize the regenerated tissue from a patient who underwent bilateral distraction and later knee replacements.MethodsKnee joint distraction was conducted using an external fixation frame and total knee arthroplasty was performed several years later. Radiographic imaging was performed to assess the status of the knee joint prior, during and after clinical interventions. Following total knee replacement, cartilage biopsies were collected and processed for tissue sectioning and histochemical staining. Tandem mass-spectrometry proteomics analysis was used to characterize and compare the proteomic composition.ResultsBoth knee joints showed joint-space improvement pre- and post-knee joint distraction. Regenerated cartilage was white with an irregular surface, while native (lateral) cartilage had a yellow appearance and smooth surface. Histochemical staining showed higher Safranin-O positivity in native cartilage compared to regenerated cartilage, and differences in collagen structure. Proteomic analysis did not reveal major differences in cartilage extracellular matrix protein abundance. Bioinformatic analyses revealed enrichment in ribosomal proteins (regenerated cartilage) and RNA Polymerase II Transcription Termination (native cartilage).ConclusionHistologically, knee joint distraction-regenerated cartilage showed less glycosaminoglycans and disorganized collagen compared to native cartilage. However, mass-spectrometry has no major differences in extracellular matrix protein abundance, with proteomic clues suggesting protein translation regulation as a potential mechanism for regeneration.

Purpose (the aim of the study): Characterization of osteoarthritis (OA) patient-specific molecular processes is essential for OA endotyping. Several studies stratified OA in a number of molecular endotypes, based on transcriptomic profiles of joint tissues. While joint tissue sampling provides insight into the underlying disease mechanisms, clinical implementation of such an approach is challenging due to invasiveness and potential damage to joint tissues. Alternatively, synovial fluid (SF) aspiration is considered a less invasive approach and more suitable for diagnostic purposes. Considering the compositional complexity of synovial fluid, we aimed to stratify OA based on SF-induced differential cellular signaling patterns.

Abstract Study question Will the transcriptional profile of SSEA-1+ endometrial epithelial cells (EECs) explain their functional role, which can be explored in an in vitro endometrial organoid model? Summary answer SSEA-1+ EECs demonstrate differentially expressed genes (DEGs) and associated functional pathways expected from a basal stem/progenitor cell (SPC) population, agreeing with the in vitro experiments. What is known already Endometrial SPCs are postulated to reside in the basalis and to be responsible for the full phenotypic and functional restoration of the endometrial functionalis layer. SSEA-1+ EECs assume the postulated basalis SPC niche. Previous studies demonstrated isolated SSEA-1+ EECs to have a higher capacity to generate organoids in 3D matrix and display growth like that observed after endometrial denudation. They have lower steroid hormone expression and higher telomerase activity with longer telomere lengths, all suggesting a SPC phenotype. SSEA-1+ EECs co-express nuclear SRY-box transcription factor 9 (nSOX9) and nuclear b-catenin, suggesting an activated Wnt pathway. Study design, size, duration Endometrial samples were collected from eight pre-menopausal women attending the Liverpool Women’s Hospital for benign gynaecological procedures, aged 30-47 years. These patients were having regular menstrual cycles and had not been on any hormonal therapy for at least three months prior to enrolment. Three further endometrial samples were obtained for 3D EEC organoid culture studies. Participants/materials, setting, methods Magnetic-activated cell sorting was used to isolate SSEA-1 enriched/depleted EECs from freshly harvested endometrial samples. Microarray analysis was performed using Agilent human arrays. DEGs and pathway enrichment analysis were explored using R-studio and Ingenuity Pathway Analysis (IPA). Internal and external validation used RT-qPCR, dual-immunofluorescence, and comparison against publicly available endometrial single cell spatial transcriptomic data. Human EEC organoids models were employed to assess hormonal regulation of SSEA-1 expression using immunohistochemistry. Main results and the role of chance SSEA-1+ EECs have a distinct transcriptional profile, with 1,059 DEGs compared with SSEA-1- EECs. Pathway analysis highlighted their role in endometrial regeneration, adhesion, remodelling, and neovascularisation, enriching for pathways such as ‘extracellular matrix structural constituent’, ‘epithelial to mesenchymal transition’, ‘angiogenesis’, ‘TNFα signalling via NFKB’ and ‘Notch signalling’. IPA’s biological functions demonstrated their involvement in tissue homeostasis, tumour suppression and their more quiescent SPC phenotype, with activation of ‘organismal death’ and inhibition of ‘cell movement/migration’ and ‘cell movement/migration of tumour cell lines’. The activation of canonical pathways such as ‘PTEN signalling’ and ‘endocannabinoid cancer inhibition’ demonstrated their fine equilibrium to drive proliferation during regeneration, whilst providing protection from hyperplastic/carcinogenic transformation. Ten selected DEGs were internally validated using RT-qPCR, demonstrating concordance. Dual-immunofluorescence of gene products of DEGs MMP7, MMP26, C11orf52 and CD47 confirmed co-localisation alongside SSEA-1 in an external biological cohort of endometrial samples. External in silico validation using the Garcia-Alonso et al endometrial single-cell transcriptomics dataset, mapped significantly upregulated DEGs MMP7 and MMP26 to mainly SOX9+ and SOX9+/LGR5+ EECs. EEC organoids exposed to hormones (including oestradiol) demonstrated an induction of proliferation and higher proportion of organoids expressing SSEA-1 (46.8% vs 67%) simulating endometrial glandular regeneration process. Limitations, reasons for caution Freshly isolated/sorted EECs were obtained from women at different menstrual cycle phases. Our organoid model system only contained EECs without the stromal component, thus, was not suitable to assess progestogenic response of EECs that occur in vivo. Wider implications of the findings This study provides novel information on human SSEA-1+ EECs, suggesting their pivotal role in endometrial regeneration. Since endometrial epithelial SPCs make a vital contribution to endometrial repair and pregnancy establishment, characterising of these cells in health will allow the development of targeted novel therapies on aberrant SPCs within gynaecological disease. Trial registration number Not applicable

Purpose: Characterization of OA patient-specific molecular processes is essential for future OA precision medicine. Several studies stratified OA in a number of molecular endotypes, based on transcriptomic profiles of joint tissues (e.g. cartilage, synovium and bone). While joint tissue sampling provides insight into the underlying disease mechanisms, clinical implementation of such an approach is challenging due to invasiveness and damage to joint tissues. Alternatively, synovial fluid aspiration is considered a less invasive approach and more suitable for diagnostic purposes. Considering the compositional complexity of synovial fluid, we aimed to stratify OA based on cell-integrated synovial fluid-induced signaling patterns rather than exploring single factors.

OBJECTIVE:Proliferative verrucous leukoplakia (PVL) is a rare form of oral leukoplakia with a relatively high transformation rate resulting in oral squamous cell carcinoma (OSCC). Molecular analysis of PVL at the genome level is limited and has only identified molecular similarities between PVL and OSCC. However, the clinical profile of PVL suggests that molecular differences may be more important.STUDY DESIGN:Whole exome sequencing of 5 PVL-associated OSCC (PVL-OSCC) and paired blood samples was used to identify somatic mutations common to the tumors. Whole methylome analysis of samples from 4 PVL-associated OSCC and 3 OSCC of non-PVL origin samples was conducted to explore differential methylation.RESULTS:In contrast to conventional OSCC, PVL-associated OSCC showed infrequent TP53 mutation and altered spectra of PIK3CA and NOTCH1 mutations. Unsupervised hierarchical clustering identified 63 probes that discriminated between PVL-associated OSCC and OSCC of non-PVL origin. Differences in methylation were most significant for divalent metal ion transport, particularly calcium movement.CONCLUSIONS:Specific differences in mutation and methylation profiles between PVL-derived OSCC and OSCC of non-PVL origin suggest differences in their transformation pathways. Further studies of early PVL lesions may identify markers of transformation that are also applicable to more common oral premalignant disorders such as oral epithelial dysplasia.

BACKGROUND:Ageing, disease and malignant transformation of the skin are associated with changes in DNA methylation. So far, mostly invasive methodologies such as biopsies have been applied in collecting DNA methylation signatures. Tape stripping offers a noninvasive option for skin diagnostics. It enables the easy but robust capture of biologic material in large numbers of participants without the need for specialized medical personnel. OBJECTIVES:To design and validate a methodology for noninvasive skin sample collection using tape stripping for subsequent DNA -methylation analysis. METHODS:A total of 175 participants were recruited and provided tape-stripping samples from a sun-exposed area; 92 provided matched tape-stripping samples from a sun-protected area, and an additional 5 provided matched skin-shave biopsies from the same area. Using -enzymatic conversion and whole-genome Illumina sequencing, we generated genome-wide DNA methylation profiles that were used to evaluate the feasibility of noninvasive data acquisition, to compare with established sampling approaches and to investigate biomarker identification for age and ultraviolet (UV) exposure. RESULTS:We found that tape-stripping samples showed strong concordance in their global DNA methylation landscapes to those of conventional invasive biopsies. Moreover, we showed sample reproducibility and consistent global methylation profiles in skin tape-stripping samples collected from different areas of the body. Using matched samples from sun-protected and sun-exposed areas of the body we were able to validate the capacity of our method to capture the effects of environmental changes and ageing in a cohort covering various ages, ethnicities and skin types. We found DNA methylation changes on the skin resulting from UV exposure and identified significant age-related hypermethylation of CpG islands, with a pronounced peak effect at 50-55 years of age, including methylation changes in well-described markers of ageing. CONCLUSIONS:These data demonstrate the feasibility of using tape stripping combined with whole-genome sequencing as a noninvasive approach to measuring DNA methylation changes in the skin. In addition, they outline a viable experimental framework for the use of skin tape stripping, particularly when it is performed in large cohorts of patients to identify biomarkers of skin ageing, UV damage and, possibly, to track treatment response to therapeutic interventions.

Extracellular vesicles (EVs) contribute to osteoarthritis pathogenesis through their release into joint tissues and synovial fluid. Synovial fluid-derived EVs have the potential to be direct biomarkers in the causal pathway of disease but also enable understanding of their role in disease progression. Utilizing a temporal model of osteoarthritis, we defined the changes in matched synovial fluid and plasma-derived EV small non-coding RNA and protein cargo using sequencing and mass spectrometry. Data exploration included time series clustering, factor analysis and gene enrichment interrogation. Chondrocyte signalling was analysed using luciferase-based transcription factor activity assays. EV protein cargo appears to be more important during osteoarthritis progression than small non-coding RNAs. Cluster analysis revealed plasma-EVs represented a time-dependent response to osteoarthritis induction associated with supramolecular complexes. Clusters for synovial fluid-derived EVs were associated with initial osteoarthritis response and represented immune/inflammatory pathways. Factor analysis for plasma-derived EVs correlated with day post-induction and were primarily composed of proteins modulating lipid metabolism. Synovial fluid-derived EVs factors represented intermediate filament and supramolecular complexes reflecting tissue repair. There was a significant interaction between time and osteoarthritis for CRE, NFkB, SRE, SRF with a trend for osteoarthritis synovial fluid-derived EVs at later time points to have a more pronounced effect.

DNA methylation in the human genome is established and maintained by DNA methyltransferases (DNMTs). DNMT isoforms show differential expression by cell lineage and during development, but much remains to be elucidated about their shared and unique genomic targets. We examined changes in the epigenome following overexpression of 13 DNMT isoforms in HEK293T cells. We observed increased methylation (Δβ > 0.2) at 43,405 CpG sites, with expression of DNMT3A2, DNMTΔ3B4 and DNMTΔ3B2 associated with the greatest impact. De novo methylation occurred primarily within open sea regions and at loci with intermediate methylation levels (β: 0.2–0.6). 53% of differentially methylated loci showed specificity towards a single DNMT subfamily, primarily DNMTΔ3B and DNMT3A and 39% towards a single isoform. These loci were significantly enriched for pathways related to neuronal development (DNMTΔ3B4), calcium homeostasis (DNMTΔ3B3) and ion transport (DNMT3L). Repetitive elements did not display differential sensitivity to overexpressed DNMTs, but hypermethylation of Alu elements was associated with their evolutionary age following overexpression of DNMT3A2, DNMT3B1, DNMT3B2 and DNMT3L. Differential methylation (Δβ > 0.1) was observed at 121 of the 353 loci associated with the Horvath ‘epigenetic clock’ model of ageing, with 51 showing isoform specificity, and was associated with reduction of epigenetic age by 5–15 years following overexpression of seven isoforms. Finally, we demonstrate the potential for dietary constituents to modify epigenetic marks through isoform-specific inhibition of methylation activity. Our results provide insight into regions of the genome methylated uniquely by specific DNMT isoforms and demonstrate the potential for dietary intervention to modify the epigenome.

DNA methylation age (DNAm age) estimation is a powerful biomarker of human ageing. To date, epigenetic clocks have not been evaluated in age-related macular degeneration (AMD). Here, we perform genome-wide DNA methylation analyses in blood of AMD patients with a documented smoking history (14 AMD, 16 Normal), identifying loci of differential methylation (DML) with a relaxed p-value criterion (p ≤ 10−4). We conduct DNAm age analyses using the Horvath-multi tissue, Hannum and Skin & Blood epigenetic clocks in both blood and retinal pigment epithelium (RPE). We perform Ingenuity Pathway Analysis Causal Network Analysis (IPA CNA) on the topmost significantly differentially methylated CpG probes in blood and RPE. Results show poor performance of epigenetic clocks in RPE. Epigenetic age acceleration (EAA) was not observed in AMD. However, we observe positive EAA in blood of smokers, and in smokers with AMD. DML analysis revealed hypomethylation at cg04953735 within RPTOR (p = 6.51 × 10−5; Δβ = −11.95%). IPA CNA in the RPE also identified RPTOR as the putative master regulator, predicted to be inhibited in AMD. In conclusion, this is the first study evaluating an association of epigenetic ageing in AMD. We posit a role for RPTOR as a common master regulator of methylation changes in the RPE in AMD.

ObjectivesThe ‘hypervirulent’ variant of Klebsiella pneumoniae (hvKp) is a predominant cause of community-acquired pyogenic liver abscess in Asia, and is an emerging pathogen in Western countries. hvKp infections have demonstrated ‘metastatic’ dissemination in immunocompetent hosts, an unusual mode of infection associated with severe complications. Two cases alerted us to the possible presence of hvKp at our hospital, both involving elderly Hispanic males who presented with recurrent fever, bacteraemia, epigastric pain and liver abscesses/phlegmon, thus prompting an assessment of hvKp prevalence.MethodsA surveillance of K. pneumoniae blood, body fluid and wound isolates was conducted using real-time PCR to detect virulence-associated genes (uni-rmpA, iucA and peg344). Positive isolates were further characterized by wzi gene sequencing to determine capsular types (K-type) and by multilocus sequence typing and pulsed-field gel electrophoresis to determine strain relatedness.ResultsFour-hundred and sixty-three K. pneumoniae isolates, derived from 412 blood, 21 body fluids and 30 abdominal wound specimens, were screened over a 3-year period. Isolates included 98 multidrug-resistant strains. Eighteen isolates from 17 patients, including two from the index patient, screened positive for all three virulence genes. Sixteen of 18 positive isolates had K-types associated with hvKp, and isolates from different patients were unrelated strains, indicating likely community acquisition. Of 13 patients with significant morbidity, five died; eight patients had co-existing hepatobiliary disease, and six had diabetes mellitus.ConclusionsMultiple strains of hvKp are emerging in New York City and are associated with high mortality relative to multidrug-resistant and classical Klebsiella infections. Co-existing hepatobiliary disease appears to be a potential risk factor for these infections.

Introduction: Pulmonary hypertension (PH) is a common consequence of myxomatous mitral valve disease (MMVD). Cavalier King Charles spaniels (CKCS) are frequently affected with MMVD and appear to have different disease progression compared to other dogs. This study aimed to determine if CKCS are more likely to develop PH as a result of MMVD than dogs of other breeds. A secondary aim was to explore whether breed or PH impacted survival. Animals: A total of 187 dogs diagnosed with MMVD, 94 CKCS and 93 non-CKCS, were included in this study. Methods: This is a retrospective review of dogs with MMVD. Data were analyzed for presence of PH, congestive heart failure (CHF) and echocardiographic variables including the ratio between mitral E wave velocity (E vel) and isovolumic relaxation time (E/IVRT) and were compared between CKCS/non-CKCS and dogs with/without PH. Survival analysis was also performed. Results: American College of Veterinary Internal Medicine (ACVIM) stage (p < 0.001), CKCS (p = 0.005), left atrium-to-aortic ratio (LA/Ao) (p < 0.001), E vel (p < 0.001) and log(10)(E/IVRT) (p < 0.001) were significant at the univariate level for PH development. At the multivariate level, only ACVIM stage remained significant (p = 0.044), suggesting that worsening MMVD was the predominant determinant of PH development in this study. Pulmonary hypertension was associated with greater likelihood of CHF (p < 0.001) and death (both cardiac [p < 0.001] and all-cause mortality [p = 0.011]). Cavalier King Charles spaniels were more likely to experience cardiac death than non-CKCS (p = 0.004). Conclusions: In this study, development of PH was associated with worse MMVD, according to ACVIM stage. (C) 2019 Elsevier B.V. All rights reserved.

Background There are limited data on the effect of endoscopic mucosal resection (EMR) on changes of histopathologic diagnosis for Barrett’s esophagus (BE) patients undergoing endoscopic eradication therapy (EET); especially those without visible lesions. Aim To compare the frequency of changes of diagnosis by EMR compared with pre-EMR biopsy diagnosis for patients with and without visible lesions. Methods In this multicenter outcomes project, patients with Barrett’s-related neoplasia undergoing EET at three tertiary-care centers were included. Patients undergoing biopsies followed by EMR within six months were included. The main outcome measures were frequency of overall change of histopathologic diagnosis, change based on pre-EMR biopsy diagnosis, and change based on the presence of visible lesions. Results One-hundred and thirty-eight BE patients (low-grade dysplasia (LGD) 15 (10.9 %), high-grade dysplasia (HGD) 87 (63 %), esophageal adenocarcinoma (EAC) 36 (26.1 %)) were included; 114 (82.6 %) patients had visible lesions. EMR resulted in a change of diagnosis for 43 (31.1 %) patients (upgrade 14 (10.1 %); downgrade 29 (21 %)). For HGD patients, EMR downstaged dysplasia grade for 17 (19.5 %) cases and upstaged it to EAC for nine (10.3 %) cases. There was a change of diagnosis for 26 (29.9 %) HGD patients, irrespective of the presence or absence of visible lesions ( p = 0.76). For EAC patients, EMR downstaged dysplasia grade in 10 (27.8 %) cases. There was a change of diagnosis for 10 (27.8 %) EAC patients, irrespective of the presence or absence of endoscopically visible lesions ( p = 0.48). Conclusions EMR results in a change of diagnosis for approximately 30 % of BE patients with early neoplasia (with and without visible lesions) referred for EET.

Epigenetic changes, such as DNA methylation, make a major contribution to cancer initiation and progression. They may precede genetic alterations and may be identified in adjacent normal tissue. Global loss of methylation may occur especially early in neoplasia and in certain tumors may be influenced by available methyl donor status (assessed by folate or homocysteine levels). We evaluated the role of global loss of methylation using LINE-1 pyrosequencing and methyl donor status in the progression of Barrett's esophagus (BE) and asked whether these changes characterize the Barrett's tissue, adjacent normal tissue (cardia and squamous epithelium) or both.

BACKGROUND: Radiofrequency ablation (RFA) is an endoscopic ablation modality used to treat Barrett's esophagus (BE) with the goal of eliminating dysplasia and metaplasia.Factors associated with stricture formation or incomplete eradication of intestinal metaplasia (EIM) are poorly understood.AIM: To determine the factors associated with stricture formation or incomplete EIM.METHODS: This was a retrospective study of all patients treated with RFA for BE at a tertiary care referral center between June 2006 and November 2010.Pertinent information was extracted from medical records, including: demographics, history of BE (pre-ablation histology, duration of pre-treatment dysplasia), medication and substance use, indicators of GERD activity (symptoms, presence of erosive esophagitis), upper endoscopy findings (Prague criteria, hiatus hernia), ablation outcomes (elimination of metaplasia and dysplasia), and complications (perforation, stricture, bleeding, and hospitalization).Outcomes related to RFA were described for all patients as well as stratified by pre-ablation histology.Comparative analysis of patients with and without stricture and complete and incomplete elimination of dysplasia were performed with non-parametric tests (Fisher's exact test for categorical data, Wilcoxon rank-sum test for continuous data) to determine associated factors.RESULTS: Among 113 patients who received RFA for BE (22 low-grade dysplasia, 77 high-grade dysplasia, 14 intramucosal carcinoma), 83 (73.5%) completed treatment with 95.2% complete elimination of dysplasia and 85.5% complete EIM.Of the remaining 30 subjects, 22 had ongoing treatment, 6 were lost to follow up, one had treatment delay for antireflux surgery and 1 opted for esophagectomy.Nine patients (8.0%) experienced a treatment-related complication, including8 strictures and 1 post-procedure hemorrhage.Stricture formation was associated with receiving endoscopic mucosal resection (75.0% vs. 36.2%,p=0.05) and number of EMR sessions (mean 1.4 vs. 0.4, p=0.007).A trend toward stricture formation existed with number of focal RFA treatments (mean 3.3 vs. 2.2, p=0.09), active NSAID use (75.0% vs. 44.8%,p=0.14) and prior peptic stricture (25.0% vs. 7.6%, p=0.15).Incomplete EIM was associated with ongoing GERD symptoms (75.0% vs. 33.8%,p=0.01) while increased Prague M length had a trend toward association (mean 6.6 vs. 4.5cm, p=0.11).CONCLUSIONS: RFA at a tertiary referral center is both safe (8.0% with complications, mostly benign strictures) and efficacious (95.2% elimination of dysplasia, 85.5% elimination of intestinal metaplasia).Previous EMR is associated with stricture formation while ongoing GERD symptoms are associated with incomplete elimination of intestinal metaplasia.These findings should be considered in planning treatment protocols for patients with BE.

The diagnostic utility of EUS in patients undergoing endoscopic eradication therapy (EET) is not clear. In addition, there is limited data on whether the diagnostic characteristics of EUS differ in BE neoplasia with and without visible lesions. In a multi-center cohort of BE patients with HGD/early EAC receiving EET:- to determine the diagnostic accuracy of EUS for determining the depth of invasion (T-stage)- to compare accuracy of EUS for T-staging in patients with and without endoscopically visible lesions This is a multicenter outcomes project of a large cohort of BE patients with HGD/early EAC being treated with EET at 3 tertiary referral centers. For this study, only biopsy proven patients with HGD/early EAC undergoing EUS followed by EMR within a 3-month time frame were included. EUS was performed by five experienced endosonographers by radial and /or miniprobe (20 MHz) endosonography. The staging of the specimen obtained by EMR was considered as the gold standard for comparison. The presence of endoscopically visible lesions in the Barrett's segment was recorded. Diagnostic characteristics including sensitivity, specificity and accuracy with 95% CI of EUS compared to EMR pathology staging (gold standard) were calculated. 105 BE patients [mean age 66.9 years, 97% Caucasian, 89% males, mean BE length 4.7 cm (SD 0.3)] were included. Baseline histology on biopsies was: HGD in 70 and EAC in 35 patients; visible lesions noted in 85 (81%) patients [HGD 57/70 (81%), EAC 28/35 (80%)]. Overall staging at EUS was: Tis 66 (62%), T1a 23 (22%), T1b 14 (13%), T2 2 (3%). In patients with HGD, staging was: Tis 52 (74%), T1a 9 (13%), T1b 9 (13%) whereas for patients with EAC, staging was: Tis 14 (40%), T1a 14 (40%), T1b 5 (14%), and T2 2 (6%). Using EMR pathology staging as the gold standard, diagnostic characteristics of EUS in the entire cohort were: sensitivity 66% (47-81), specificity 75% (64-85), and accuracy 72% (63-81). Results based on biopsy pathology and presence of visible lesion has been highlighted in Table 1, Table 2. In HGD patients, EMR resulted in a change in diagnosis in 27/70 (38%) cases: upgrading to EAC in 12/70 (17%) and downgrading in 15/70 (21%). Similarly, in EAC patients, EMR resulted in a change in diagnosis in 15/35 (43%) cases: downgrading to HGD in 12/35 (34%) and LGD/IM in 3/35 (9%).Table 1Diagnostic characteristics of EUS based on baseline histologyBiopsy HGDBiopsy EACp valueSensitivity (95% CI)33% (10-65)85% (62-97)0.006Specificity (95% CI)76% (63-86)73% (45-92)1Accuracy (95% CI)69% (56-79)80% (63-91)0.25 Open table in a new tab Table 2Diagnostic characteristics of EUS based on endoscopically visible lesionsVisible lesion presentVisible lesion absentp valueSensitivity (95% CI)65% (46-82)67% (10-99)1Specificity (95% CI)71% (58-83)87% (62-98)0.32Accuracy (95% CI)69% (58-79)84% (60-97)0.26 Open table in a new tab Accuracy of EUS for T-staging of BE patients with HGD/early EAC is limited. Initial biopsy pathology and presence of visible lesions did not affect the overall accuracy of EUS. The role of EUS in BE with early neoplasia should be reevaluated and its overall impact on patient management should be assessed in future prospective trials.