Modeling mechanisms underlying differential inflammatory responses to COVID-19 in type 2 diabetes using a patient-derived microphysiological organ-on-a-chip system.

Background: COVID-19 pandemic has caused more than 6 million deaths worldwide. Co-morbid conditions such as Type 2 Diabetes (T2D) have increased mortality in COVID-19. With limited translatability of in vitro and small animal models to human disease, human organ-on-a-chip models are an attractive platform to model in vivo disease conditions and test potential therapeutics. Methods: T2D or non-diabetic patient-derived macrophages and human liver sinusoidal endothelial cells were seeded, along with normal hepatocytes and stellate cells in the liver-on-a-chip (LAMPS - liver acinus micro physiological system), perfused with media mimicking non-diabetic fasting or T2D (high levels of glucose, fatty acids, insulin, glucagon) states. The macrophages and endothelial cells were transduced to overexpress the SARS-CoV2-S (spike) protein with appropriate controls before their incorporation into LAMPS. Cytokine concentrations in the efflux served as a read-out of the effects of S-protein expression in the different experimental conditions (non-diabetic-LAMPS, T2D-LAMPS), including incubation with tocilizumab, an FDA-approved drug for severe COVID-19. Findings: S-protein expression in the non-diabetic LAMPS led to increased cytokines, but in the T2D-LAMPS, this was significantly amplified both in the number and magnitude of key pro-inflammatory cytokines (IL6, CCL3, IL1ß, IL2, TNFα, etc.) involved in cytokine storm syndrome (CSS), mimicking severe COVID-19 infection in T2D patients. Compared to vehicle control, tocilizumab (IL6-receptor antagonist) decreased the pro-inflammatory cytokine secretion in T2D-COVID-19-LAMPS but not in non-diabetic-COVID-19-LAMPS. Interpretation: macrophages and endothelial cells play a synergistic role in the pathophysiology of the hyper-inflammatory response seen with COVID-19 and T2D. The effect of Tocilizumab was consistent with large clinical trials that demonstrated Tocilizumab's efficacy only in critically ill patients with severe disease, providing confirmatory evidence that the T2D-COVID-19-LAMPS is a robust platform to model human in vivo pathophysiology of COVID-19 in T2D and for screening potential therapeutics.

Light-Dark Patterns Mirroring Shift Work Accelerate Atherosclerosis and Promote Vulnerable Lesion Phenotypes.

Background Despite compelling epidemiological evidence that circadian disruption inherent to long-term shift work enhances atherosclerosis progression and vascular events, the underlying mechanisms remain poorly understood. A challenge to the use of mouse models for mechanistic and interventional studies involving light-dark patterns is that the spectral and absolute sensitivities of the murine and human circadian systems are very different, and light stimuli in nocturnal mice should be scaled to represent the sensitivities of the human circadian system. Methods and Results We used calibrated devices to deliver to low-density lipoprotein receptor knockout mice light-dark patterns representative of that experienced by humans working day shifts or rotating shift schedules. Mice under day shifts were maintained under regular 12 hours of light and 12 hours of dark cycles. Mice under rotating shift schedules were subjected for 11 weeks to reversed light-dark patterns 4 days in a row per week, followed by 3 days of regular light-dark patterns. In both protocols the light phases consisted of monochromatic green light at an irradiance of 4 µW/cm2. We found that the shift work paradigm disrupts the foam cell's molecular clock and increases endoplasmic reticulum stress and apoptosis. Lesions of mice under rotating shift schedules were larger and contained less prostabilizing fibrillar collagen and significantly increased areas of necrosis. Conclusions Low-density lipoprotein receptor knockout mice under light-dark patterns analogous to that experienced by rotating shift workers develop larger and more vulnerable plaques and may represent a valuable model for further mechanistic and/or interventional studies against the deleterious vascular effects of rotating shift work.

Differentiation of extrahepatic bile duct cholangiocarcinoma from benign stricture: findings at MRCP versus ERCP.

PURPOSE: To retrospectively evaluate criteria for differentiating extrahepatic bile duct cholangiocarcinoma from benign cause of stricture at magnetic resonance cholangiopancreatography (MRCP) and to compare diagnostic accuracy with this modality versus endoscopic retrograde cholangiopancreatography (ERCP). MATERIALS AND METHODS: MRCP and ERCP images in 50 patients (27 with cholangiocarcinoma [18 men, nine women; mean age, 58 years] and 23 with benign cause of stricture [13 men, 10 women; mean age, 60 years]) were retrospectively reviewed to assess the appearance of bile duct strictures. Final diagnosis was based on surgical or biopsy findings. Strictures were described according to their imaging appearance (irregular or smooth margins, asymmetric or symmetric narrowing, abrupt narrowing or gradual tapering, and presence or absence of double-duct sign). Sensitivity, specificity, and accuracy of MRCP and ERCP were calculated by using ratings of confidence in image-based diagnosis. Lengths of stricture were electronically measured and compared by using the Student t test. RESULTS: Among cholangiographic criteria for malignant biliary stricture, irregular margins and asymmetric narrowing were more common in cholangiocarcinomas (24 [89%] of 27 patients) than in benign strictures (six [26%] and eight [35%] of 23 patients, respectively). Sensitivity, specificity, and accuracy of the two methods for differentiation of malignant from benign causes of biliary stricture were 81% (22 of 27), 70% (16 of 23), and 76% (38 of 50), respectively, for MRCP and 74% (20 of 27), 70% (16 of 23), and 72% (36 of 50), respectively, for ERCP. Mean length (+/- standard deviation) of cholangiocarcinomas was 30.0 mm +/- 8.5, and that of benign strictures was 13.6 mm +/- 9.1 (P <.001). CONCLUSION: Accuracy of MRCP is comparable with that of ERCP. Regardless of modality, a lengthy segment of extrahepatic bile duct stricture with irregular margin and asymmetric narrowing suggests cholangiocarcinoma, and a short segment with regular margin and symmetric narrowing suggests benign cause.

Scirrhous gastric carcinoma: endoscopy versus upper gastrointestinal radiography.

PURPOSE: To compare the accuracy of upper gastrointestinal (UGI) series and endoscopic examination in the diagnosis and localization of scirrhous gastric carcinoma. MATERIALS AND METHODS: Seventy-two patients with pathologically proved scirrhous gastric carcinoma in surgical specimens were included. Preoperative reports at UGI series and endoscopic examination, which included impressions on the location and extent of the tumor, were compared with pathology reports, and the accuracy of the preoperative reports was calculated. Two gastrointestinal radiologists retrospectively reviewed the appearance of mucosa at UGI series. RESULTS: Preoperative diagnoses at endoscopy were Borrmann type IV carcinoma in 28 patients (39%), type III carcinoma in 29 (40%), early gastric carcinoma in seven (10%), lymphoma in six (8%), atrophic gastritis in one (3%), and type II carcinoma in one (3%). Preoperative diagnoses at UGI series were type IV carcinoma in 44 patients (61%), type III carcinoma in 25 (35%), lymphoma in two (3%), and early gastric carcinoma in one (1%). Pathology reports were compared with the preoperative reports, and tumor location and extent were correct in the endoscopic examination reports of 24 patients (33%) and the UGI series reports of 49 patients (68%). In 68 patients, UGI series revealed thickened and irregular folds in 62 (91%), ulceration in 42 (62%), and nodularity in 22 (32%) at consensus review. Endoscopic biopsy samples were positive for malignancy in 66 patients (93%). CONCLUSION: UGI series is superior to endoscopic examination in the diagnosis and localization of scirrhous gastric carcinoma.