Faculty Bibliography

Isorhamnetin is an O-methylated flavonol present in fruit and vegetables. We recently reported the identification of isorhamnetin as an activator of the human pregnane X receptor (PXR), a known target for abrogating inflammation in inflammatory bowel disease (IBD). The current study investigated the role of isorhamnetin as a putative mouse PXR activator in ameliorating chemically induced IBD. Using two different models (ulcerative colitis like and Crohn's disease like) of experimental IBD in mice, we demonstrated that isorhamnetin abrogated inflammation through inhibiting the activity of myeloperoxidase, the levels of TNF-alpha and IL-6, the mRNA expression of proinflammatory mediators (iNOS, ICAM-1, COX2, TNF-alpha, IL-2 and IL-6) and the phosphorylation of IkappaBalpha and NF-kappaB p65. PXR gene overexpression inhibited NF-kappaB luciferase activity, and the inhibition was potentiated by isorhamnetin treatment. PXR knockdown by siRNA demonstrated the necessity for PXR in isorhamnetin-mediated up-regulation of xenobiotic metabolism genes. Ligand pocket-filling mutants (S247W/C284W and S247W/C284W/S208W) of human PXR weakened the effect of isorhamnetin on PXR activation. Molecular docking studies and time-resolved fluorescence resonance energy transfer competitive binding assays confirmed the ligand (isorhamnetin)-binding affinity. These results clearly demonstrated the ameliorating effect of isorhamnetin on experimental IBD via PXR-mediated up-regulation of xenobiotic metabolism and down-regulation of NF-kappaB signaling. The novel findings may contribute to the effective utilization of isorhamnetin or its derivatives as a PXR ligand in the treatment of human IBD.

Intestinal microbial metabolites are conjectured to affect mucosal integrity through an incompletely characterized mechanism. Here we showed that microbial-specific indoles regulated intestinal barrier function through the xenobiotic sensor, pregnane X receptor (PXR). Indole 3-propionic acid (IPA), in the context of indole, is a ligand for PXR in vivo, and IPA downregulated enterocyte TNF-alpha while it upregulated junctional protein-coding mRNAs. PXR-deficient (Nr1i2(-/-)) mice showed a distinctly "leaky" gut physiology coupled with upregulation of the Toll-like receptor (TLR) signaling pathway. These defects in the epithelial barrier were corrected in Nr1i2(-/-)Tlr4(-/-) mice. Our results demonstrate that a direct chemical communication between the intestinal symbionts and PXR regulates mucosal integrity through a pathway that involves luminal sensing and signaling by TLR4.

Primary hepatic signet ring cell neuroendocrine tumor is extremely rare and is characterized by distinct intracytoplasmic hyaline vacuoles that are mucin negative and cytokeratin positive. The unique histological features may cause difficulty in diagnosis and delay patient care. Here the authors report a 49-year-old man with an incidental finding of a 2.7 cm liver mass in the absence of chronic liver disease. The resected tumor was grossly unencapsulated but well demarcated with friable tissue texture. Microscopically, the entire tumor consisted of sheets of monotonous cells separated by delicate microvasculature. The tumor cells had granular chromatin, inconspicuous nucleoli, and eosinophilic cytoplasm. Many of the tumor cells had eccentric, pale intracytoplasmic vacuoles resembling signet ring cells in adenocarcinoma. Immunohistochemical studies showed that the tumor cells were positive for neuroendocrine markers and that the intracytoplasmic vacuoles were negative for mucin but strongly positive for cytokeratins. Careful systemic search including OctreoScan scintigraphy (Mallinckrodt Medical, Inc., St. Louis, MO) and capsule endoscopy failed to reveal any other tumors. A diagnosis of primary hepatic signet ring cell neuroendocrine tumor was established. Ten months after surgery, the patient is well without any other detectable tumor on radiology. Serological neuroendocrine markers are also within normal limits.

We subjected 23 formalin-fixed, paraffin-embedded tissue blocks (11 cases of thymic hyperplasia and 12 thymomas [3 encapsulated, 8 with capsular invasion, and 1 atypical]) to incubation with monoclonal anti-X-linked inhibitor of apoptosis protein (XIAP) (BD Biosciences, San Jose, CA) and monoclonal anti-p63 (4A4, Santa Cruz, Santa Cruz, CA). Granular or heterogeneous cytoplasmic XIAP staining and nuclear p63 staining were considered positive. We compared thymic hyperplasia with thymoma and capsulated thymoma with thymoma with capsular invasion or atypia. p63 was positive in virtually all thymic epithelial cells in hyperplasia and thymoma. XIAP was negative in all hyperplasia cases except one. Of 12 thymomas, 9 were XIAP+ with focal/weak to diffuse/strong positivity: 2 of 3 encapsulated and 7 of 8 thymomas with capsular invasion were XIAP+. One atypical thymoma was XIAP-. XIAP expression differed significantly between hyperplasia and thymoma (P = .0007) but not between capsulated and invasive thymomas (P = .3797). p63 is consistently positive in nonneoplastic and neoplastic thymic epithelium. XIAP expression in thymoma suggests a possible role in the pathogenesis of thymoma and may be helpful in differentiating thymic hyperplasia from thymoma, especially in small biopsy specimens. However, the level of expression does not correlate with capsular invasion or atypia.

CONTEXT: Clear cell carcinoma of the urinary bladder/ urethra is a rare tumor histologically resembling the neoplasms in the female genital tract. Adequate characterization of this tumor has been hampered by its rarity. alpha-Methylacyl-CoA racemase (AMACR)/P504S has been reported to be positive in prostatic adenocarcinoma, papillary renal cell carcinoma, and gastrointestinal neoplasmas; however, it has never been studied in clear cell carcinoma of the lower urinary tract. OBJECTIVE: To investigate the immunohistochemical staining profile in 4 primary clear cell carcinomas of the urinary tract, including P504S, which has not been previously evaluated in these tumors. DESIGN: Four cases of clear cell adenocarcinoma were retrieved from our archives: 2 cases from the urinary bladder (one each from a man and a woman) and 2 cases from the urethra (both from women, 1 in a diverticulum). Immunohistochemistry performed on the cases were P504S, K903, cytokeratin (CK) 7, CK20, CA 125, and p63. RESULTS: We found that clear cell carcinomas had a distinct immunoreactive profile: strongly positive for P504S, K903, and CK7, and negative for p63. Two cases were also positive for CA 125 and CK20. CONCLUSION: The immunohistochemical profile of clear cell carcinomas shares some similarity to conventional urothelial carcinoma; however, it deviates from those tumors in being positive for P504S and negative for p63. This staining profile may suggest a nonurothelial origin for these tumors, may serve as a useful tool in the differential diagnosis of this tumor, and may reflect its etiology. Because similar expression of P504S is also seen in nephrogenic adenomas, this marker should not be used to differentiate nephrogenic adenomas from clear cell adenocarcinomas.