Cutaneous Keratocyst With D2-40 Immunoreactivity in Basal Cell Nevus Syndrome.

ABSTRACT: Although not a diagnostic criterion for basal cell nevus syndrome (BCNS, OMIM#109400), cutaneous cysts, particularly epidermoid cysts, are common in this condition. Cutaneous keratocysts, on the other hand, are extremely rare in general and have been identified in only 5 patients with BCNS. Here, we describe a BCNS patient with a cutaneous keratocyst that demonstrated D2-40 (podoplanin) immunoreactivity, which has been detected in odontogenic keratocysts but not cutaneous keratocysts. This finding suggests that cutaneous keratocysts may be developmentally homologous to odontogenic keratocysts and may behave similarly in terms of invasion and growth pattern.

Malignant Glomus Tumor Arising in a Long Standing Precursor Lesion.

Malignant glomus tumor is an exceedingly rare vascular malignancy of unclear etiology, which is believed to arise de novo or from malignant transformation of a benign glomus tumor. The authors present a case of an elderly woman evaluated for a painful 2.5-cm purple-brown tumor on the right forearm that grew rapidly within a long-standing, seemingly benign lesion. An excisional biopsy revealed a multilobular dermal tumor composed of areas of benign-appearing glomus tumor cells interspersed with areas of dense, pleomorphic cells arranged in sheets and fascicles with an elevated mitotic rate. The diagnosis of malignant glomus tumor was made on the basis of the lesion's nuclear atypia and elevated mitotic rate. This case demonstrates malignant transformation of a presumably benign glomus tumor precursor lesion highlighting this rarely reported phenomenon. The authors present this case to raise awareness of this vascular malignancy that carries a significant risk of metastasis.

Amorphous basophilic deposit in the superficial dermis of the lip in an 80 year old.

An 80-year-old woman presented with a 3 mm pearly, translucent papule in her left upper cutaneous lip of 2 months duration which was biopsied to rule out basal cell carcinoma. Histopathologic examination revealed acellular, basophilic material in the superficial dermis, extending to the base of the biopsy. There was neither an epithelial component nor an inflammatory reaction associated with it. The amorphous, nonpolarizable material stained with Alcian Blue (pH 2.5) and with colloidal iron, but was negative with a Periodic acid-Schiff stain, indicative of an acidic mucin, such as weakly sulfated mucin of salivary gland origin or a dermal-origin mucin. The material was digested with hyaluronidase, consistent with the mesenchymal-origin mucin hyaluronic acid (HA). Additional clinical history was obtained; the patient had previous HA (Restylane) injections at another institution. We report a case of superficially applied HA and consider the histopathologic differential diagnosis of endogenous and injected mucin in the dermis.

Value of glypican 3 immunostaining in the diagnosis of hepatocellular carcinoma on needle biopsy.

The diagnostic value of glypican 3 (GPC3) immunostaining on needle biopsy specimens has not been well assessed. In this study, 120 liver needle biopsy specimens, including 46 from cirrhotic livers and 74 hepatocellular carcinomas (HCCs), were immunohistochemically examined for expression of GPC3. The results showed strong cytoplasmic and membranous staining in 36 HCCs (49%), among which 20 cases (56%) showed diffuse immunoreactivity. None of the 46 cirrhotic livers exhibited positive GPC3 immunostaining. The nonneoplastic liver tissues (cirrhotic or noncirrhotic) that were present in the majority of the HCC cases were also completely negative for GPC3 expression. These data demonstrate that GPC3 is a reliable immunohistochemical marker for the diagnosis of HCC on needle biopsy specimens when positive. However, the detection rate in our series seems lower than that reported in studies using resection specimens as the study materials. Our findings emphasize that GPC3 immunoreactivity can be focal and that negative staining should not be viewed as evidence to exclude the diagnosis of HCC in challenging needle biopsy specimens.

Photosensitizer delivery to vulnerable atherosclerotic plaque: comparison of macrophage-targeted conjugate versus free chlorin(e6).

We have previously shown that a conjugate (MA-ce6) between maleylated serum albumin and the photosensitizer chlorin(e6) (ce6) is targeted in vitro to macrophages via class A scavenger receptors. We now report on the ability of this conjugate to localize in macrophage-rich atherosclerotic plaques in vivo. Both the conjugate and the free photosensitizer ce6 are studied after injection into New Zealand White rabbits that are rendered atherosclerotic by a combination of aortic endothelial injury and cholesterol feeding into normal rabbits. Rabbits are sacrificed at 6 and 24 h after injection and intravascular fluorescence spectroscopy is carried out by fiber-based fluorimetry in intact blood-filled arteries. Surface spectrofluorimetry of numbered excised aortic segments together with injured and normal iliac arteries is carried out, and quantified ce6 content by subsequent extraction and quantitative fluorescence determination of the arterial segments and also of nontarget organs. There is good agreement between the various techniques for quantifying ce6 localization, and high contrast between arteries from atherosclerotic and normal rabbits is obtained. Fluorescence correlates with the highest burden of plaque in the aorta and the injured iliac artery. The highest accumulation in plaques is obtained using MA-ce6 at 24 h. Free ce6 gives better accumulation at 6 h compared to 24 h. The liver, spleen, lung, and gall bladder have the highest uptake in nontarget organs. Macrophage-targeted photosensitizer conjugates may have applications in both detecting and treating inflamed vulnerable plaque.

Primary peritoneal epithelioid hemangioendothelioma.

Epithelioid hemangioendothelioma is an uncommon malignant vascular tumor of intermediate grade that occurs in a variety of organs and soft tissues. Diagnosis of epithelioid hemangioendothelioma is often complicated by the rarity of the tumor, and because the tumor shares many morphologic features with other peritoneal neoplasms. This report presents 3 cases of epithelioid hemangioendotheliomas arising as primary tumors of the peritoneum and reviews 7 previously reported cases to establish the demographic, clinical, morphologic, immunohistochemical, and ultrastructural features of this neoplasm. These cases demonstrate that the light microscopic features are very similar to epithelioid hemangioendothelioma arising at more conventional sites, such as the liver, that immunohistochemical analysis provides a reliable approach for confirming or establishing the diagnosis, and that at least one endothelial marker (either CD31, CD34, or factor VIII) should be positive for a definitive diagnosis. This series identifies the characteristics of peritoneal epithelioid hemangioendothelioma that can be reliably used for diagnosis.

Glypican-3 as a useful diagnostic marker that distinguishes hepatocellular carcinoma from benign hepatocellular mass lesions.

CONTEXT: Histopathologic distinction between hepatocellular carcinoma (HCC) and benign hepatocellular mass lesions, particularly hepatocellular adenoma, can sometimes be challenging. The currently available ancillary tools are suboptimal in terms of sensitivity and specificity. OBJECTIVE: To further characterize the diagnostic value of glypican-3 (GPC3), a cell surface proteoglycan that has recently been shown to be overexpressed in HCC, in the distinction between HCC and benign hepatocellular mass lesions. DESIGN: A total of 221 surgically resected liver specimens were subjected to immunohistochemical staining using a monoclonal antibody specific for GPC3. These included 111 HCCs, 48 hepatocellular adenomas, 30 focal nodular hyperplasias, and 32 large regenerative nodules in the background of cirrhosis. RESULTS: Cytoplasmic, membranous, and canalicular staining for GPC3 was detected in 84 (75.7%) of the 111 HCCs, among which, 61 (72.6%) of the 84 cases exhibited diffuse immunoreactivity. In contrast, none of the 110 cases of hepatocellular adenoma, focal nodular hyperplasia, and large regenerative nodule showed detectable GPC3 staining. Focal GPC3 immunoreactivity was detected in cirrhotic nodules in 11 (16.4%) of 67 HCC cases with a cirrhotic background, but no background staining was observed in the remaining 44 HCCs without cirrhosis. GPC3 expression in HCCs did not correlate with the size, differentiation, or stage of the tumors; the presence or absence of cirrhotic background; or the underlying etiologies. CONCLUSIONS: GPC3 is a specific immunomarker for HCC that can be used to distinguish HCC from benign hepatocellular mass lesions, particularly hepatocellular adenoma. However, the diagnosis of HCC should not rely entirely on positive GPC3 immunostaining because focal immunoreactivity can be detected in a small subset of cirrhotic nodules. In addition, GPC3 expression in HCC can also be focal, and thus, the lack of GPC3 staining does not exclude the diagnosis of HCC.

Intravascular detection of inflamed atherosclerotic plaques using a fluorescent photosensitizer targeted to the scavenger receptor.

Inflammation plays an important role in the pathophysiology of atherosclerotic disease. We have previously shown that the targeted photosensitizer chlorin (e(6)) conjugated with maleylated albumin (MA-ce6) is taken up by macrophages via the scavenger receptor with high selectivity. In a rabbit model of inflamed plaque in New Zealand white rabbits via balloon injury of the aorto-iliac arteries and high cholesterol diet we showed that the targeted conjugate showed specificity towards plaques compared to free ce6. We now show that an intravascular fiber-based spectrofluorimeter advanced along the -iliac vessel through blood detects 24-fold higher fluorescence in atherosclerotic vessels compared to control rabbits (p < 0.001 ANOVA). Within the same animals, signal derived from the injured iliac artery was 16-fold higher than the contralateral uninjured iliac (p < 0.001). Arteries were removed and selective accumulation of MA-ce6 in plaques was confirmed using: (1) surface spectrofluorimetry, (2) fluorescence extraction of ce6 from aortic segments, and (3) confocal microscopy. Immunohistochemical analysis of the specimens showed a significant correlation between MA-ce6 uptake and RAM-11 macrophage staining (R = 0.83, p < 0.001) and an inverse correlation between MA-ce6 uptake and smooth muscle cell staining (R = -0.74, p < 0.001). MA-ce6 may function as a molecular imaging agent to detect and/or photodynamically treat inflamed plaques.

Macrophage-targeted photosensitizer conjugate delivered by intratumoral injection.

A conjugate between maleylated albumin and a photosensitizer (PS) shows cell type specific targeting to macrophages via the scavenger receptor. Administration of this conjugate to a tumor-bearing mouse followed by illumination may allow selective destruction of macrophages within tumors. There is accumulating evidence that tumor-associated macrophages contribute to tumor growth, invasiveness, metastasis, and immune suppression. We tested the intravenous (IV) injection of a conjugate between maleylated albumin and chlorin(e6) to BALB/c mice bearing three tumor types with differing proportions of tumor-associated macrophages. The accumulation of PS within the tumors after IV injection and 24 h incubation time was disappointing, and we therefore investigated intratumoral (IT) injection. This gave 20-50 times greater concentrations of PS within the tumor compared to IV injection as determined by tissue extraction. Furthermore the amounts of PS in each tumor type correlated well with the numbers of macrophages both as determined by extraction from bulk tumor and fluorescence quantification and as determined by tissue dissociation to a single cell suspension and two-color flow cytometry with macrophage-specific antibodies. IT injection of nonconjugated PS gave lower tumor accumulation that did not correlate with macrophage content. IT injection of targeted macromolecular delivery systems is an underexplored area and worthy of further study.

Healing of perforating rat corneal incisions closed with photodynamic laser-activated tissue glue.

BACKGROUND AND OBJECTIVES: Laser-activated photodynamic biologic tissue glues may be useful for closing incisions in ophthalmology. We report on the use of two such preparations to close perforating corneal incisions in living rats. STUDY DESIGN/MATERIALS AND METHODS: A previously described preparation containing a covalent albumin-chlorin e6 (ce6) conjugate (bovine serum albumin (BSA)-ce6), and a novel mixture of albumin and Janus Green (BSA/JG), both activated with a 665-nm diode laser were used to glue mouse skin ex vivo. The optimized glues were then used to seal incisions in rat corneas and results were compared to control incisions. Rats were sacrificed at day 1, 7, and 14 and eyes tested for leaking pressure and examined histopathologically. RESULTS: One day after treatment eyes closed with BSA-ce6 had a leaking pressure (in mmHg) of 357 compared to 193 for control incisions (P<0.01); closure with BSA/JG gave a leaking pressure of 430 (P<0.05 compared to BSA-ce6, and P<0.001 compared to control). Histological examination showed eyes sealed with BSA/JG have less inflammation present than untreated eyes at 7 days. CONCLUSIONS: These data demonstrate that photodynamic laser activated tissue glues can be used to effectively seal corneal incisions in living animals without thermal damage or undue inflammation.