SUMMARYThe unsteady incompressible Navier-Stokes equations are formulated in terms of vorticity and streamfunction in generalized curvilinear orthogonal co-ordinates to facilitate analysis of flow configurations with general geometries. The numerical method developed solves the conservative form of the vorticity transport equation using the alternating direction implicit method, whereas the streamfunction equation is solved by direct block Gaussian elimination. The method is applied to a model problem of flow over a backstep in a doubly infinite channel, using clustered conformal co-ordinates. One-dimensional stretching functions, dependent on the Reynolds number and the asymptotic behaviour of the flow, are used to provide suitable grid distribution in the separation and reattachment regions, as well as in the inflow and outflow regions. The optimum grid distribution selected attempts to honour the multiple length scales of the separated flow model problem. The asymptotic behaviour of the finite differenced transport equation near infinity is examined and the numerical method is carefully developed so as to lead to spatially second-order-accurate wiggle-free solutions, i.e. with minimum dispersive error. Results have been obtained in the entire laminar range for the backstep channel and are in good agreement with the available experimental data for this flow problem, prior to the onset of three-dimensionality in the experiment.

Background: Eosinophilic gastritis (EG) is a clinicopathologic disorder with marked gastric eosinophilia and clinical symptoms. There is an unmet need among patients with EG for more precise diagnostic tools. Objective: We aimed to develop tissue-and blood-based diagnostic platforms for EG. Methods: Patients with EG and control subjects without EG were enrolled across 9 Consortium of Eosinophilic Gastrointestinal Disease Researchers-associated sites. An EG Diagnostic Panel (EGDP; gastric transcript subset) and EG blood biomarker panel (protein multiplex array) were analyzed. EGDP 18 scores were derived from the expression of 18 highly dysregulated genes, and blood EG scores were derived from dysregulated cytokine/chemokine levels. Results: Gastric biopsy specimens and blood samples from 185 subjects (patients with EG, n 5 74; control subjects without EG, n 5 111) were analyzed. The EGDP (1) identified patients with active EG (P < .0001, area under the curve > _ 0.95), (2) effectively monitored disease activity in longitudinal samples (P 5 .0078), (3) highly correlated in same-patient samples (antrum vs body, r 5 0.85, P < .0001), and (4) inversely correlated with gastric peak eosinophil levels (r 5 20.83, P < .0001), periglandular circumferential collars (r 5 20.73, P < .0001), and endoscopic nodularity (r 5 20.45, P < .0001). For blood-based platforms, eotaxin-3, thymus and activation-regulated chemokine, IL-5, and thymic stromal lymphopoietin levels were significantly increased. Blood EG scores (1) distinguished patients with EG from control subjects without EG (P < .0001, area under the curve > _ 0.91), (2) correlated with gastric eosinophil levels

Eosinophilic esophagitis (EoE) is a chronic, food antigen–driven, inflammatory disease of the esophagus and is associated with impaired barrier function. Evidence is emerging that loss of esophageal expression of the serine peptidase inhibitor, kazal type 7 (SPINK7), is an upstream event in EoE pathogenesis. Here, we provide evidence that loss of SPINK7 mediates its pro-EoE effects via kallikrein 5 (KLK5) and its substrate, protease-activated receptor 2 (PAR2). Overexpression of KLK5 in differentiated esophageal epithelial cells recapitulated the effect of SPINK7 gene silencing, including barrier impairment and loss of desmoglein-1 expression. Conversely, KLK5 deficiency attenuated allergen-induced esophageal protease activity, modified commensal microbiome composition, and attenuated eosinophilia in a murine model of EoE. Inhibition of PAR2 blunted the cytokine production associated with loss of SPINK7 in epithelial cells and attenuated the allergen-induced esophageal eosinophilia in vivo. Clinical samples substantiated dysregulated PAR2 expression in the esophagus of patients with EoE, and delivery of the clinically approved drug α1 antitrypsin (A1AT, a protease inhibitor) inhibited experimental EoE. These findings demonstrate a role for the balance between KLK5 and protease inhibitors in the esophagus and highlight EoE as a protease-mediated disease. We suggest that antagonizing KLK5 and/or PAR2 has potential to be therapeutic for EoE.