Objective-Tissue factor pathway inhibitor (TFPI) is an endothelial-associated inhibitor of blood coagulation. Because the mechanism for attachment of TFPI to endothelium is not clear, we investigated its association with human placenta. Methods and Results-Western blots demonstrate that treatment with phosphatidylinositol-specific phospholipase C (PIPLC) removes more placental TFPI than either PBS or heparin, a finding confirmed by immunohistochemistry. The amounts of heparin-releasable and PIPLC-releasable TFPI activity on placental endothelium were measured in placentas from 5 individuals. PIPLC removes Ͼ10-fold more TFPI activity from the placental fragments than 10 U/mL heparin and Ͼ100-fold more than 1 U/mL heparin. Pretreatment of the placental fragments with PIPLC increases the amount of heparin-releasable TFPI by Ϸ3-fold. An antibody specific for the C-terminal region of TFPI recognizes PIPLC-releasable TFPI in Western blots. Conclusions-GPI-anchored TFPI is the predominant form on placental endothelium. Heparin-releasable TFPI likely represents only a small portion of the total TFPI on endothelium that remains attached to cell-surface glycosaminoglycans after cleavage of the GPI anchor by endogenous enzymes. The predominance of GPI-anchored TFPI suggests that heparin infusion does not significantly redistribute TFPI within the vasculature. The intact C-terminus in GPI-anchored TFPI indicates it is not directly attached to a GPI anchor. Rather, it most likely associates with endothelium by binding to a GPI-anchored protein.

The distribution of the 52 kDa keratin 13 was evaluated immunohistochemically, using the AE8 monoclonal antibody. Various squamous and transitional cell epithelial lesions and representative control tissues were studied. This antibody performed adequately in formalin-fixed and paraffin-embedded tissue, but like keratin immunohistochemistry in general, required protease pretreatment. Keratin 13 was found consistently in the suprabasal layers of squamous epithelia of oral cavity, tonsils, larynx, esophagus, lower female genital tract, and transitional urothelium, but it was absent in the epidermis. Generally, various forms of squamous metaplasia were AE8-positive. In dysplasia, AE8 reactivity was considerably decreased or even absent despite the presence of apparent suprabasal maturation. In differentiated squamous cell carcinomas, AE8 immunoreactivity was usually limited to a few cells in the center of the keratinized foci. However, in 10% of squamous cell carcinomas, a significant number of tumor cells was positive. Only well-differentiated urothelial carcinomas showed AE8 immunoreactivity, while poorly differentiated tumors were negative. Interestingly, a Brenner's tumor showed a high number of AE8-positive epithelial cells. Our results show that the expression of keratin 13, as immunohistochemically determined by AE8 antibody, is significantly down-regulated in squamous cell malignancies. Its possible value as an adjunct to diagnosis of dysplasia should be investigated further.

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Inducible nitric oxide synthase is expressed in inflammatory macrophages in areas of chronic inflammation in the bladder mucosa of spinal cord injured patients with a chronic indwelling bladder catheter. The expression of inducible nitric oxide synthase may potentially lead to the sustained production of nitric oxide and its oxidative products, the nitrosation of urinary amines and the formation of potentially carcinogenic nitrosamines in the bladder.

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