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PURPOSE: This study uses deep neural network-generated rim-to-disc area ratio (RADAR) measurements and the disc damage likelihood scale (DDLS) to measure the rate of optic disc rim loss in a large cohort of glaucoma patients. METHODS: A deep neural network was used to calculate RADAR and DDLS for each optic disc photograph (ODP). Patient demographics, diagnosis, intraocular pressure (IOP), and mean deviation (MD) from perimetry were analyzed as risk factors for faster progression of RADAR. Receiver operating characteristic (ROC) curves were used to compare RADAR and DDLS in their utility to distinguish glaucoma from glaucoma suspect (GS) and for detecting glaucoma progression. RESULTS: A total of 13,679 ODPs with evidence of glaucomatous optic nerve damage from 4106 eyes of 2407 patients with glaucoma or GS were included. Of these eyes, 3264 (79.5%) had a diagnosis of glaucoma, and 842 (20.5%) eyes were GS. Mean ± SD baseline RADAR of GS and glaucoma were 0.67 ± 0.13 and 0.57 ± 0.18, respectively (P < 0.001). Older age, greater IOP fluctuation, baseline MD, right eye, and diagnosis of secondary open-angle glaucoma were associated with slope of RADAR. The mean baseline DDLS of GS and glaucoma were 3.78 and 4.39, respectively. Both RADAR and DDLS showed a less steep slope in advanced glaucoma. In glaucoma, the change of RADAR and DDLS correlated with the corresponding change in MD. RADAR and DDLS had a similar ability to discriminate glaucoma from GS and detect disease progression. Area under the ROC curve of RADAR and DDLS was 0.658 and 0.648. CONCLUSIONS: Automated calculation of RADAR and DDLS with a neural network can be used to evaluate the extent and long-term rate of optic disc rim loss and is further evidence of long-term nerve fiber loss in treated patients with glaucoma. TRANSLATIONAL RELEVANCE: Our study provides a large clinic-based experience for RADAR and DDLS measurements in GS and glaucoma with a neural network.

AIM: We tested the hypothesis that visual field (VF) progression can be predicted with a deep learning model based on longitudinal pairs of optic disc photographs (ODP) acquired at earlier time points during follow-up. METHODS: 3919 eyes (2259 patients) with ≥2 ODPs at least 2 years apart, and ≥5 24-2 VF exams spanning ≥3 years of follow-up were included. Serial VF mean deviation (MD) rates of change were estimated starting at the fifth visit and subsequently by adding visits until final visit. VF progression was defined as a statistically significant negative slope at two consecutive visits and final visit. We built a twin-neural network with ResNet50-backbone. A pair of ODPs acquired up to a year before the VF progression date or the last VF in non-progressing eyes were included as input. Primary outcome measures were area under the receiver operating characteristic curve (AUC) and model accuracy. RESULTS: The average (SD) follow-up time and baseline VF MD were 8.1 (4.8) years and -3.3 (4.9) dB, respectively. VF progression was identified in 761 eyes (19%). The median (IQR) time to progression in progressing eyes was 7.3 (4.5-11.1) years. The AUC and accuracy for predicting VF progression were 0.862 (0.812-0.913) and 80.0% (73.9%-84.6%). When only fast-progressing eyes were considered (MD rate < -1.0 dB/year), AUC increased to 0.926 (0.857-0.994). CONCLUSIONS: A deep learning model can predict subsequent glaucoma progression from longitudinal ODPs with clinically relevant accuracy. This model may be implemented, after validation, for predicting glaucoma progression in the clinical setting.

PurposeTo develop a patient-reported outcome measure to assess the impact of glaucoma and treatment, including minimally invasive glaucoma surgery (MIGS).DesignObservational study before and after concomitant cataract and Food and Drug Administration-approved implantable MIGS device surgery.SettingSurvey administration was on a computer, iPad, or similar device.Patient population184 adults completed the baseline survey, 124 a survey 3 months after surgery, and 106 the 1-month test-retest reliability survey. The age range was 37 to 89 (average age = 72). Most were female (57%), non-Hispanic White (81%), and had a college degree (56%).Main outcome measuresThe Glaucoma Outcomes Survey (GOS) assesses functional limitations (27 items), vision-related symptoms (7 items), psychosocial issues (7 items), and satisfaction with microinvasive glaucoma surgery (1 item). These multiple-item scales were scored on a 0 to 100 range, with a higher score indicating worse health.ResultsInternal consistency reliability estimates ranged from 0.75 to 0.93, and 1-month test-retest intraclass correlations ranged from 0.83 to 0.92 for the GOS scales. Product-moment correlations among the scales ranged from 0.56 to 0.60. Improvement in visual acuity in the study eye from baseline to the 3-month follow-up was significantly related to improvements in GOS functional limitations (r = 0.18, P = .0485), vision-related symptoms (r = 0.19, P = .0386), and psychosocial concerns (r = 0.18, P = .0503). Responders to treatment ranged from 17% for vision-related symptoms to 48% for functional limitations.ConclusionsThis study supports using the GOS for ophthalmic procedures such as MIGS. Further evaluation of the GOS in different patient subgroups and clinical settings is needed.

Qualitative secondary analysis has generated heated debate regarding the epistemology of qualitative research. We argue that shifting to an abductive approach provides a fruitful avenue for qualitative secondary analysts who are oriented towards theory-building. However, the concrete implementation of abduction remains underdeveloped--especially for coding. We address this key gap by outlining a set of tactics for abductive analysis that can be applied for qualitative analysis. Our approach applies Timmermans and Tavory's (Timmermans and Tavory 2012; Tavory and Timmermans 2014) three stages of abduction in three steps for qualitative (secondary) analysis: Generating an Abductive Codebook, Abductive Data Reduction through Code Equations, and In-Depth Abductive Qualitative Analysis. A key contribution of our article is the development of "code equations"--defined as the combination of codes to operationalize phenomena that span individual codes. Code equations are an important resource for abduction and other qualitative approaches that leverage qualitative data to build theory.

Case summary Ectopic thyroid tissue is rarely reported in dogs and cats in its prediaphragmatic location and has never been described in the liver. A 15-year-old spayed female domestic shorthair cat was diagnosed by ultrasound with a heterogeneous hypoechoic nodular area in the liver at the periphery of the quadrate lobe. A generic diagnosis of carcinoma was made after ultrasound-guided fine-needle aspiration and cytological examination. The patient underwent staging by CT scan and subsequently underwent hepatic lobectomy. Histologically, a diagnosis of thyroid adenocarcinoma was made, confirmed immunohistochemically using positive thyroglobulin staining; the tumour was suspected to be of metastatic origin. CT scans excluded primary thyroid involvement; in addition, lesions at other sites were not detected. Therefore, a final diagnosis of thyroid adenocarcinoma arising from ectopic thyroid tissue in the liver was made. The cat recovered uneventfully from surgery. Relevance and novel information This report describes an unusual case of an adenocarcinoma originating from presumed thyroid ectopic tissue within the liver of a cat. Ectopic thyroid tissue has been rarely reported in both dogs and cats and, to the authors’ knowledge, it has never been described in the liver of a cat.

PURPOSE: Demonstrate that a novel Bayesian hierarchical spatial longitudinal (HSL) model improves estimation of local macular ganglion cell complex (GCC) rates of change compared to simple linear regression (SLR) and a conditional autoregressive (CAR) model. METHODS: We analyzed GCC thickness measurements within 49 macular superpixels in 111 eyes (111 patients) with four or more macular optical coherence tomography scans and two or more years of follow-up. We compared superpixel-patient-specific estimates and their posterior variances derived from the latest version of a recently developed Bayesian HSL model, CAR, and SLR. We performed a simulation study to compare the accuracy of intercept and slope estimates in individual superpixels. RESULTS: HSL identified a significantly higher proportion of significant negative slopes in 13/49 superpixels and a significantly lower proportion of significant positive slopes in 21/49 superpixels than SLR. In the simulation study, the median (tenth, ninetieth percentile) ratio of mean squared error of SLR [CAR] over HSL for intercepts and slopes were 1.91 (1.23, 2.75) [1.51 (1.05, 2.20)] and 3.25 (1.40, 10.14) [2.36 (1.17, 5.56)], respectively. CONCLUSIONS: A novel Bayesian HSL model improves estimation accuracy of patient-specific local GCC rates of change. The proposed model is more than twice as efficient as SLR for estimating superpixel-patient slopes and identifies a higher proportion of deteriorating superpixels than SLR while minimizing false-positive detection rates. TRANSLATIONAL RELEVANCE: The proposed HSL model can be used to model macular structural measurements to detect individual glaucoma progression earlier and more efficiently in clinical and research settings.

PURPOSE: To evaluate and compare the effectiveness of nearest neighbor (NN)- and variational autoencoder (VAE)-smoothing algorithms to reduce variability and enhance the performance of glaucoma visual field (VF) progression models. DESIGN: Longitudinal cohort study. SUBJECTS: 7150 eyes (4232 patients), with ≥ 5 years of follow-up and ≥ 6 visits. METHODS: Vsual field thresholds were smoothed with the NN and VAE algorithms. The mean total deviation (mTD) and VF index rates, pointwise linear regression (PLR), permutation of PLR (PoPLR), and the glaucoma rate index were applied to the unsmoothed and smoothed data. MAIN OUTCOME MEASURES: The proportion of progressing eyes and the conversion to progression were compared between the smoothed and unsmoothed data. A simulation series of noiseless VFs with various patterns of glaucoma damage was used to evaluate the specificity of the smoothing models. RESULTS: The mean values of age and follow-up time were 62.8 (standard deviation: 12.6) years and 10.4 (standard deviation: 4.7) years, respectively. The proportion of progression was significantly higher for the NN and VAE smoothed data compared with the unsmoothed data. VF progression occurred significantly earlier with both smoothed data compared with unsmoothed data based on mTD rates, PLR, and PoPLR methods. The ability to detect the progressing eyes was similar for the unsmoothed and smoothed data in the simulation data. CONCLUSIONS: Smoothing VF data with NN and VAE algorithms improves the signal-to-noise ratio for detection of change, results in earlier detection of VF progression, and could help monitor glaucoma progression more effectively in the clinical setting. FINANCIAL DISCLOSURES: Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article.

Isocitrate Dehydrogenase-1 (IDH1) is commonly mutated in lower-grade diffuse gliomas. The IDH1R132H mutation is an important diagnostic tool for tumor diagnosis and prognosis; however, its role in glioma development, and its impact on response to therapy, is not fully understood. We developed a murine model of proneural IDH1R132H-mutated glioma that shows elevated production of 2-hydroxyglutarate (2-HG) and increased trimethylation of lysine residue K27 on histone H3 (H3K27me3) compared to IDH1 wild-type tumors. We found that using Tazemetostat to inhibit the methyltransferase for H3K27, Enhancer of Zeste 2 (EZH2), reduced H3K27me3 levels and increased acetylation on H3K27. We also found that, although the histone deacetylase inhibitor (HDACi) Panobinostat was less cytotoxic in IDH1R132H-mutated cells (either isolated from murine glioma or oligodendrocyte progenitor cells infected in vitro with a retrovirus expressing IDH1R132H) compared to IDH1-wild-type cells, combination treatment with Tazemetostat is synergistic in both mutant and wild-type models. These findings indicate a novel therapeutic strategy for IDH1-mutated gliomas that targets the specific epigenetic alteration in these tumors.

BACKGROUND: Which phenotypes are we able to recognize in the optic nerve of patients with primary open angle glaucoma? METHODS: Retrospective interventional case series. 885 eyes from 885 patients at an outpatient tertiary care centre who met specified criteria for POAG were included. Disc photographs were classified by three glaucoma specialists into the following phenotypes according to their predominant characteristics: (1) concentric rim thinning, (2) focal rim thinning, (3) acquired pit of the optic nerve (APON), (4) tilted, (5) extensive peripapillary atrophy (PPA), and (6) broad rim thinning. Demographic, medical, and ocular data were collected. Kruskal-Wallis was used as a non-parametric test and pairwise comparison was performed by using Wilcoxon rank sum test corrected. RESULTS: Phenotypic distribution was as follows: 398(45%) focal thinning, 153(18%) concentric thinning, 153(17%) broad thinning, 109(12%) tilted, 47(5%) extensive PPA and 25(3%) APON. Phenotypic traits of interest included a higher proportion of female patients with the focal thinning phenotype (p = 0.015); myopia (p = 0.000), Asian race (OR: 8.8, p = 0.000), and younger age (p = 0.000) were associated with the tilted phenotype; the concentric thinning patients had thicker RNFL (p = 0.000), higher MD (p = 0.008) and lower PSD (p = 0.043) than broad thinning, despite no difference in disc sizes (p = 0.849). The focal thinning group had a localized VF pattern with high PSD compared to concentric thinning (p = 0.005). CONCLUSION: We report six phenotypic classifications of POAG patients with demographic and ocular differences between phenotypes. Future refinement of phenotypes should allow enhanced identification of genetic associations and improved individualization of patient care.

PURPOSE: Predict central 10° global and local visual field (VF) measurements from macular optical coherence tomography (OCT) volume scans with deep learning (DL). METHODS: This study included 1121 OCT volume scans and 10-2 VFs from 289 eyes (257 patients). Macular scans were used to estimate 10-2 VF mean deviation (MD), threshold sensitivities (TS), and total deviation (TD) values at 68 locations. A three-dimensional (3D) convolutional neural network based on the 3D DenseNet121 architecture was used for prediction. We compared DL predictions to those from baseline linear models. We carried out 10-fold stratified cross-validation to optimize generalizability. The performance of the DL and baseline models was compared based on correlations between ground truth and predicted VF measures and mean absolute error (MAE; ground truth - predicted values). RESULTS: Average (SD) MD was -9.3 (7.7) dB. Average (SD) correlations between predicted and ground truth MD and MD MAE were 0.74 (0.09) and 3.5 (0.4) dB, respectively. Estimation accuracy deteriorated with worsening MD. Average (SD) Pearson correlations between predicted and ground truth TS and MAEs for DL and baseline model were 0.71 (0.05) and 0.52 (0.05) (P < 0.001) and 6.5 (0.6) and 7.5 (0.5) dB (P < 0.001), respectively. For TD, correlation (SD) and MAE (SD) for DL and baseline models were 0.69 (0.02) and 0.48 (0.05) (P < 0.001) and 6.1 (0.5) and 7.8 (0.5) dB (P < 0.001), respectively. CONCLUSIONS: Macular OCT volume scans can be used to predict global central VF parameters with clinically relevant accuracy. TRANSLATIONAL RELEVANCE: Macular OCT imaging may be used to confirm and supplement central VF findings using deep learning.

The objective of this study was to identify the structural guiding elements for the implementation of a telecommuting model in the post-pandemic scenario. The theoretical framework was based on specialized literature on telecommuting and organizational change, particularly drawing from the design structure proposed in Galbraith's Star Model. The research strategy is a case study type. The data analysis technique used was content analysis of interviews conducted with 24 leaders and employees of the cooperative related to the topic, generating 409 citations, 58 subcategories, and 5 analytical categories, which were systematized with the help of the software Atlas.ti 8.0. After data analysis, it was evident that the main structural (synthetic) elements for the implementation of the work model are consistent with the dimensions of the chosen theoretical framework: strategy, structure, processes and lateral capability, reward system, and people system. However, it was also possible to note that there are specific and significant structural (analytical) elements that need to be considered by organizations that aim to migrate to or implement a telecommuting model, considering the peculiarities of the specific context of the cooperative organization in this study.

PURPOSE: To compare rates of change (RoC) of peripapillary retinal nerve fiber layer (RNFL) and Bruch membrane opening-based minimum rim width (BMO-MRW) thickness in moderate-to-advanced glaucoma. DESIGN: Prospective cohort study. METHODS: Longitudinal optical coherence tomography (OCT) optic nerve head volume scans of 113 eyes of 113 glaucoma patients with moderate-to-advanced or central damage were exported. This study estimated and compared global and sectoral RoC with linear mixed effects models and simple linear regression (SLR) of RNFL and BMO-MRW thickness. Permutation analyses were used to test significance of RoC in the SLR model. It also compared longitudinal signal-to-noise ratios (LSNR) defined as RoC divided by residual standard deviation (SD) between the two groups. RESULTS: Mean (SD) follow-up and median (IQR) OCT scan sessions were 5.2 (1.3) years and 10 (8-11), respectively. Baseline average (SD) visual field mean deviation was -9.2 (5.8) dB. Based on SLR, a higher proportion of significant negative RNFL RoC was observed compared to BMO-MRW in the inferotemporal (35% vs 20%; P = .015) and inferonasal (42% vs 17%; P < .001) sectors. Permutation analyses also demonstrated a higher proportion of worsening RNFL RoC than BMO-MRW in the inferotemporal (P = .026) and inferonasal (P < .001) sectors along with overall lower positive RoC. Longitudinal signal-to-noise ratios for RNFL were significantly more negative than for BMO-MRW globally, and in the inferotemporal, inferonasal, and superonasal sectors (P ≤ .01). CONCLUSIONS: Longitudinal RNFL OCT measurements are more likely to detect structural change and demonstrate better LSNR compared with BMO-MRW in eyes with central or moderate-to-advanced glaucoma damage at baseline.

Sustainability in the fruit industry and protection of local cultivars are nowadays important tasks for consumers and authorities. Phenolic compounds are interesting to explore because they play an important role in fruit-related health benefits. The phenolic profile of the regional and commercial Spanish apple cultivars was performed by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). For a deeper understanding of their anti-inflammatory and antioxidant potential, the 5-lipoxigenase inhibition and NO− scavenging were evaluated. Moreover, cellular viability was determined to perform MTT assay in HepG2 and HeLa cell lines to discard cytotoxicity. The results showed that procyanidin B2 (PB2), 3-caffeoylquinic acid, epicatechin and 4-hydroxy benzoic acid were the most abundant phenolic compounds. Autochthonous apple cultivars showed better 5-LOX/NO− inhibition and nontoxic effects in the tested cell lines with an interesting profile of phenolic compounds. Local autochthonous apples can be considered a better source of bioactive compounds than the commercial samples.

Abstract Celtis australis L. (Family: Cannabaceae) is commonly used to treat many diseases like gastrointestinal problems, menstrual bleeding and amenorrhea. The present study was designed to investigate the chemical constituents, antioxidant, enzyme inhibitory and cytotoxic properties of different extracts from twigs, fruits and leaves of C. australis. EtOAc, EtOH, 70% EtOH and aqueous extracts were prepared by maceration. Results showed that the EtOH extract of the leaves had the highest total phenolic content and possessed remarkable antiradical, ion reducing and total antioxidant activities. Additionally, the leaves (EtOH or EtOAc extracts) exerted the best enzyme inhibition properties. The polar extracts of the leaves had significant cytotoxic effect against the human colorectal adenocarcinoma (HT‐29) and human prostate cancer (DU‐145) cell lines while the EtOAc of the twigs was effective against the former cell line. Phytochemically, the twigs and fruits accumulated high content of vanillic acid, 4‐hydroxy benzoic acid and syringic acid. Through a combination of in vitro and in silico approaches, we identified key phytochemicals exhibited significant inhibitory effects on several cancer‐related proteins, through in vitro and in silico approaches that show significant inhibition of cancer‐related proteins. In conclusion, these findings indicated that C. australis could be a promising source of bioactive molecules for food, pharmaceutical and cosmetic industries.

BackgroundThe phosphodiesterase 4 (PDE4) inhibitor apremilast downregulates the production of IL-23 and other pro-inflammatory cytokines involved in the pathogenesis of psoriatic arthritis (PsA).AimTo investigate the effects of apremilast on the production of cytokines by peripheral blood monocyte-derived macrophages, innate-like lymphocyte cells (ILCs), mucosal-associated invariant T (MAIT) cells, γδ T cells, natural killer (NK) cells, and NKT-like cells from patients with PsA manifesting different clinical responses to the treatment.MethodsPeripheral blood samples were obtained from patients with PsA at baseline and after 1 and 4 months of apremilast therapy (n = 23) and 20 controls with osteoarthritis. Cytokine expression in peripheral blood monocyte-derived macrophages and ILCs/MAIT/γδT/NK/NKT-like cells was tested by RT-PCR and FACS analyses, respectively; cytokine levels in culture supernatants and sera were analyzed by ELISA.ResultsPsA monocyte-derived macrophages exhibited higher expressions of IL-23, IL-1β, and TNF-α, compared with OA controls, more profoundly in patients responding to apremilast. There were 17/23 (74%) PsA patients who were classified as responders to apremilast at 4 months, and a baseline serum IL-23 >1.4 pg/mL was associated with the responder status (AUCROC 0.79; sensitivity 100%, specificity 68%). Of note, apremilast led to a significantly reduced expression of IL-23 in peripheral blood monocyte-derived macrophages; IL-17 in ILC1 and in T cells of responder patients; IFN-γ in γδ T lymphocytes.ConclusionAn enhanced myeloid inflammatory signature characterizes PsA monocyte-derived macrophages, and serum IL-23 levels represent candidate biomarkers for PsA response to apremilast.

Colorectal cancer (CRC) is a multifaceted disease whose development and progression varies depending on tumor location, age of patients, infiltration of immune cells within cancer lesions, and the tumor microenvironment. These pathophysiological characteristics are additionally influenced by sex-related differences. The gut microbiome plays a role in initiation and progression of CRC, and shapes anti-tumor immune responses but how responsiveness of the immune system to the intestinal microbiota may contribute to sexual dimorphism of CRC is largely unknown. We studied survival, tumor-infiltrating immune cell populations and tumor-associated microbiome of a cohort of n = 184 male and female CRC patients through high-dimensional single-cell flow cytometry and 16S rRNA gene sequencing. We functionally tested the immune system-microbiome interactions in in-vivo and in-vitro models of the disease. High-dimensional single-cell flow cytometry showed that female patients are enriched by tumor-infiltrating invariant Natural Killer T (iNKT) cells but depleted by cytotoxic T lymphocytes. The enrichment of oral pathobionts and a reduction of β-glucuronidase activity are distinctive traits characterizing the gut microbiome of female patients affected by CRC. Functional assays using a collection of human primary iNKT cell lines demonstrated that the gut microbiota of female patients functionally impairs iNKT cell anti-tumor functions interfering with the granzyme-perforin cytotoxic pathway. Our results highlight a sex-dependent functional relationship between the gut microbiome, estrogen metabolism, and the decline of cytotoxic T cell responses, contributing to the sexual dimorphism observed in CRC patients with relevant implications for precision medicine and the design of targeted therapeutic approaches addressing sex bias in cancer.

The prickly pear (Opuntia ficus-indica L., Cactaceae) seeds can be used as an alternative food product by the torrefaction process. Torrefaction is a pre-treatment process of biomass into solid fuel within a temperature range of 60–200 °C. This study aimed to investigate the effect of torrefaction parameters on the antioxidant potential of prickly pear seeds, which is considered a food by-product, by using the Central Composite Design approach (CCD).The best torrefaction parameters found to have the optimal phytochemical contents were: 200 °C and 50 min with 104.86 ± 1.94 GAE/g extract for Total Phenolic Content (TPC), 200 °C and 50 min with 81.23 ± 0.90 mg QE/g extract for Total Flavonoid Content (TFC), 200 °C and 50 min with IC50 = 90.66 ± 2.09 μg/ml for free radical scavenging activity (DPPH assay), 200 °C and 50 min with IC50 = 124.9 ± 4 μg/ml for radical cation total antioxidant capacity (ABTS assay). Our results demonstrated an increase of antioxidant activities with the increase of torrefaction parameters. Therefore, torrefaction has proven to be a value-added process to exploit prickly pear seed biomass and use it as a source of antioxidant food additives.

The interaction between the gut microbiota and invariant Natural Killer T (iNKT) cells plays a pivotal role in colorectal cancer (CRC). The pathobiont Fusobacterium nucleatum influences the anti-tumor functions of CRC-infiltrating iNKT cells. However, the impact of other bacteria associated with CRC, like Porphyromonas gingivalis, on their activation status remains unexplored. In this study, we demonstrate that mucosa-associated P. gingivalis induces a protumour phenotype in iNKT cells, subsequently influencing the composition of mononuclear-phagocyte cells within the tumor microenvironment. Mechanistically, in vivo and in vitro experiments showed that P. gingivalis reduces the cytotoxic functions of iNKT cells, hampering the iNKT cell lytic machinery through increased expression of chitinase 3-like-1 protein (CHI3L1). Neutralization of CHI3L1 effectively restores iNKT cell cytotoxic functions suggesting a therapeutic potential to reactivate iNKT cell-mediated antitumour immunity. In conclusion, our data demonstrate how P. gingivalis accelerates CRC progression by inducing the upregulation of CHI3L1 in iNKT cells, thus impairing their cytotoxic functions and promoting host tumor immune evasion.

Mucosal enrichment of the Adherent-Invasive E. coli (AIEC) pathotype and the expansion of pathogenic IFNγ-producing Th17 (pTh17) cells have been linked to Crohn’s Disease (CD) pathogenesis. However, the molecular pathways underlying the AIEC-dependent pTh17 cell transdifferentiation in CD patients remain elusive. To this aim, we created and functionally screened a transposon AIEC mutant library of 10.058 mutants to identify the virulence determinants directly implicated in triggering IL-23 production and pTh17 cell generation. pTh17 cell transdifferentiation was assessed in functional assays by co-culturing AIEC-infected human dendritic cells (DCs) with autologous conventional Th17 (cTh17) cells isolated from blood of Healthy Donors (HD) or CD patients. AIEC triggered IL-23 hypersecretion and transdifferentiation of cTh17 into pTh17 cells selectively through the interaction with CD-derived DCs. Moreover, the chronic release of IL-23 by AIEC-colonized DCs required a continuous IL-23 neutralization to significantly reduce the AIEC-dependent pTh17 cell differentiation. The multi-step screenings of the AIEC mutant’s library revealed that deletion of ybaT or rfaP efficiently hinder the IL-23 hypersecretion and hampered the AIEC-dependent skewing of protective cTh17 into pathogenic IFNγ-producing pTh17 cells. Overall, our findings indicate that ybaT (inner membrane transport protein) and rfaP (LPS-core heptose kinase) represent novel and attractive candidate targets to prevent chronic intestinal inflammation in CD.

Hepatitis E virus (HEV) is recognized as an emerging zoonosis. Pigs and wild boars are considered the main reservoirs of zoonotic HEV-3 and HEV-4 genotypes. In Europe, autochthonous human cases of hepatitis E, mainly associated with HEV-3 and consumption of raw or undercooked pig and wild boar liver/meat, have increased over the last decades. From 2016 to 2024, during several hunting seasons, we conducted a molecular and serological longitudinal survey on the circulation of HEV in Maremman wild boar (Italian subspecies/ecotype, Sus scrofa majori) and fallow deer (Dama dama) populations in a protected area in Central Italy. During the study period, 346 livers (256 from wild boar, 90 from fallow deer), 161 serum (127 from wild boar, 34 from fallow deer), and 23 meat juice (11 from wild boar, 12 from fallow deer) samples were collected. Serum and meat juice samples were tested using a commercial ELISA test for the detection of total anti-HEV antibodies. An estimated serological prevalence of 28.3% (39/138) in wild boar and 21.7% (10/46) in fallow deer was found. The 346 liver samples were tested using a HEV Real-Time RT-PCR for the detection of HEV-RNA. Thirty-one wild boar (12%) and four fallow deer (4.4%) livers were found positive. Phylogenetic analysis of 11 partial ORF2 sequences from wild boar confirmed the HEV3 heterogeneity in this species, revealing different strains (3f, 3c) circulating over the years. The detected subtypes are among the most commonly detected in Italy and our strains showed a high correlation with human and wild boar Italian strains. Although the studied area is a fenced natural reserve, the presence of different strains over time suggests the probable virus introduction from the external. Our results confirm fallow deer susceptibility to the infection, and that wild boar could be considered the main wild HEV reservoir. This is also the first study demonstrating the infection in the so-called Italian subspecies/ecotype Maremman wild boar. Moreover, our results corroborate that the consumption of undercooked or raw liver from both wild boar and fallow deer, or the direct contact with these animals, could represent a zoonotic risk.

K2 performance depends on different kinematic and kinetic variables. Due to the lack of related studies in this area, we have tried to explain these features to better understand the best positioning of paddlers and how their synchronization affects performance. This study uses the DAQ system comprising two instrumented paddles—an IMU and a GPS (“E-kayak” system)—to investigate paddle synchronization and the specific positioning of paddlers’ in preferred and inverted configurations. In this study, 10 sub-elite paddlers participated, divided into five crews. The test included two trials of 500 m performed in preferred and inverted seating positions. The synchronization analysis highlighted that the rear paddler contributed efficiently to the propulsion of the boat while performing 30–40 ms earlier than the front paddler during the entry and exit phases. Despite the time results for 500 m, there is no evidence indicating a dominant indication of the preferred or inverted position among the athletes. The results show a significant correlation (p < 0.05) between the force of the front paddler (r = −0.88), the stroke frequency of the crew (r = −0.66), and the total force applied by the crew with the time for 500 m and between stroke frequency and the force of the front paddler (r = 0.64). Based on these indications, for only those crews who completed the 500 m test in the shortest time, the equation determining the time over 500 m was calculated using multiple regression analysis, considering the stroke frequency and the force of the front and rear paddler. The data showed a good estimation with CV% = 0.22, ICC = 0.99, and ES = −0.005. In conclusion, these findings can serve as a beneficial tool for assessing or monitoring K2 crew performance in sub-elite paddlers.