Acquisition of antigen-presenting functions by neutrophils isolated from mice with chronic colitis.

Active episodes of the inflammatory bowel diseases are associated with the infiltration of large numbers of myeloid cells including neutrophils, monocytes, and macrophages. The objective of this study was to systematically characterize and define the different populations of myeloid cells generated in a mouse model of chronic gut inflammation. Using the T cell transfer model of chronic colitis, we found that induction of disease was associated with enhanced production of myelopoietic cytokines (IL-17 and G-CSF), increased production of neutrophils and monocytes, and infiltration of large numbers of myeloid cells into the mesenteric lymph nodes (MLNs) and colon. Detailed characterization of these myeloid cells revealed three major populations including Mac-1(+)Ly6C(high)Gr-1(low/neg) cells (monocytes), Mac-1(+)Ly6C(int)Gr-1(+) cells (neutrophils), and Mac-1(+)Ly6C(low/neg)Gr-1(low/neg) leukocytes (macrophages, dendritic cells, and eosinophils). In addition, we observed enhanced surface expression of MHC class II and CD86 on neutrophils isolated from the inflamed colon when compared with neutrophils obtained from the blood, the MLNs, and the spleen of colitic mice. Furthermore, we found that colonic neutrophils had acquired APC function that enabled these granulocytes to induce proliferation of OVA-specific CD4(+) T cells in an Ag- and MHC class II-dependent manner. Finally, we observed a synergistic increase in proinflammatory cytokine and chemokine production following coculture of T cells with neutrophils in vitro. Taken together, our data suggest that extravasated neutrophils acquire APC function within the inflamed bowel where they may perpetuate chronic gut inflammation by inducing T cell activation and proliferation as well as by enhancing production of proinflammatory mediators.

Capturing fine-grained details for video-based automation of suturing skills assessment.

OBJECTIVES: Manually-collected suturing technical skill scores are strong predictors of continence recovery after robotic radical prostatectomy. Herein, we automate suturing technical skill scoring through computer vision (CV) methods as a scalable method to provide feedback. METHODS: Twenty-two surgeons completed a suturing exercise three times on the Mimic™ Flex VR simulator. Instrument kinematic data (XYZ coordinates of each instrument and pose) were captured at 30 Hz. After standardized training, three human raters manually video segmented suturing task into four sub-stitch phases (Needle handling, Needle targeting, Needle driving, Needle withdrawal) and labeled the corresponding technical skill domains (Needle positioning, Needle entry, Needle driving, and Needle withdrawal). The CV framework extracted RGB features and optical flow frames using a pre-trained AlexNet. Additional CV strategies including auxiliary supervision (using kinematic data during training only) and attention mechanisms were implemented to improve performance. RESULTS: This study included data from 15 expert surgeons (median caseload 300 [IQR 165-750]) and 7 training surgeons (0 [IQR 0-8]). In all, 226 virtual sutures were captured. Automated assessments for Needle positioning performed best with the simplest approach (1 s video; AUC 0.749). Remaining skill domains exhibited improvements with the implementation of auxiliary supervision and attention mechanisms when deployed separately (AUC 0.604-0.794). All techniques combined produced the best performance, particularly for Needle driving and Needle withdrawal (AUC 0.959 and 0.879, respectively). CONCLUSIONS: This study demonstrated the best performance of automated suturing technical skills assessment to date using advanced CV techniques. Future work will determine if a "human in the loop" is necessary to verify surgeon evaluations.

Papillary oncocytic cystadenoma of the parotid glands: a report of 2 cases with varied cytologic features.

BACKGROUND: Papillary cystadenoma is a rare salivary gland neoplasm, and oncocytic change can be focal or marked. Papillary oncocytic cystadenoma has been reported mainly in the minor salivary glands and occasionally in the parotid glands. The cytologic features vary. CASES: A 26-year-old female presented with a 2.4-cm, cystic right parotid gland mass present for 4 months. Fine needle aspiration (FNA) showed cellular smears with sheets of oncocytic cells and some with micropapillary architecture. The diagnosis ofa neoplasm was rendered and excisional biopsy recommended. Oncocytic neoplasm was diagnosed during intraoperative consultation, and final surgical histology showed a unilocular, cystic, oncocytic neoplasm with variable papillary projections. A 52-year-old male presented with a 1.5-cm, cystic left parotid mass enlarging for the past few months. FNA showed less cellular smears with extensive necrotic debris and a few large sheets of epithelial cells with vague papillary architecture. Oncocytic neoplasm was diagnosed during intraoperative consultation, and final surgical histology showed a unilocular cystic lesion with multiple papillary fronds lined with oncocytic cells and focal metaplastic squamous cells. CONCLUSION: Papillary cystadenoma is rare in the parotid glands, and cytologic features may vary. Warthin's tumor, oncocytoma, intraductal papilloma and acinic cell carcinoma may arise in the differential diagnosis. In cases with extensive necrotic debris and metaplastic squamous cells, branchial cyst and cystic metastatic squamous carcinoma may also need to be considered.

Fine needle aspiration of salivary glands: 5-year experience from a single academic center.

OBJECTIVE: To investigate 5 years' experience with fine needle aspiration (FNA) of salivary glands at a single academic center. STUDY DESIGN: A total of 191 salivary gland FNAs were performed at Louisiana State University Health Science Center from 2003 to 2007, and all were done on major salivary glands except for 1 case. RESULTS: The cytologic diagnoses included 17 malignancies, 6 atypia, 73 neoplasms, 87 negative and 18 nondiagnostic. Eighty-six cases had histologic follow-up (45.0%). There were 5 false negatives: 2 adenoid cystic carcinomas, 1 acinic cell carcinoma, 1 polymorphous low grade adenocarcinoma and 1 metastatic basaloid squamous cell carcinoma. The only false positive was a pleomorphic adenoma misdiagnosed as adenoid cystic carcinoma. Four reactive processes were diagnosed as benign neoplasms, including 2 granulomatous inflammation and 2 chronic sialadenitis. Five benign neoplasms were interpreted as reactive processes, including 2 Warthin's tumors, 2 sebaceous lymphoadenomas and 1 pleomorphic adenoma. The overall accuracy in distinguishing benign from malignant lesions was 79.1%, and the sensitivity for salivary neoplasia was 89.4%. CONCLUSION: Our results are consistent with the literature that salivary gland FNA has good sensitivity, specificity and accuracy in the diagnosis of salivary neoplasms. FNA can play a significant role in triaging patients with onsite cytologic interpretation and can reduce many unnecessary surgeries.

Roles of T cell-associated L-selectin and ß7 integrins during induction and regulation of chronic colitis.

BACKGROUND: L-selectin (CD62L) and ß(7) integrins are important for trafficking of naive T cells under steady-state conditions. The objectives of this study were to dissect the requirements for T cell-associated CD62L and ß(7) integrins during initiation, progression, and regulation of chronic colitis. METHODS: Using the T-cell transfer model, we compared colitogenic potential between T cells lacking one or both of these molecules with wild-type T cells. To assess trafficking of cells to the secondary lymphoid tissue and the gut, we performed co-homing experiments. RESULTS: Adoptive transfer of wild-type, CD62L(-/-) or ß(7)(-/-) single-deficient T cells induced moderate to severe disease with slightly different kinetics. However, transfer of CD62L(-/-) ß(7)(-/-) double-deficient (DKO) T cells produced significantly attenuated gut inflammation, which correlated with fewer T cells and reduced levels of proinflammatory cytokines in the colon lamina propria. Our subsequent experiments established that lack of colitogenic potential of these cells was due to inability of DKO T cells to home to the secondary lymphoid tissue. Furthermore, homing of in vitro-generated effector DKO T cells to the inflamed intestine was significantly impaired. Lastly, DKO regulatory T cells were ineffective at suppressing colitis induced by wild-type T cells. CONCLUSIONS: We established that T cells can use either CD62L(-/-) or ß(7)(-/-) integrins to induce chronic colitis, but lack of both abrogates their colitogenic potential. Effector T cells critically rely on ß(7) integrin during their recruitment to the inflamed intestinal mucosa. Finally, regulation of intestinal inflammation by regulatory T cells requires one or both of these adhesion molecules.