A new class of soluble basic protein precursors of the cornified envelope of mammalian epidermis.

The cornified envelope has been shown to be formed beneath the plasma membrane as a result of the cross-linking of soluble and membrane-associated precursor proteins by transglutaminase. We have obtained a monoclonal antibody which reacts with the periphery of cells in the upper layers of human epidermis by indirect immunofluorescence (IIF) following immunization of mice with cornified envelopes of cultured human keratinocytes. The antibody also stained the cell peripheries of bovine, rat and mouse epidermis as well as stratified epithelium. Neutral buffer extracts of human cultured keratinocytes and epidermis examined under denaturing conditions contained polypeptides of molecular weight 14,900 and 16,800 which reacted with the antibody, and an additional component of molecular weight 24,800 was found in cultured cells. The polypeptides were shown to have a pI of about 9.0. Under non-denaturing conditions the two lower-molecular-weight polypeptides had an apparent molecular weight of 30,000, while the 24,800 protein had one of 60,000. Incubation of the polypeptides under conditions that activate transglutaminase resulted in a disappearance of the polypeptides or the formation of cross-linked products. Basic polypeptides with somewhat different pI values and molecular weights were identified in neutral buffer extracts of bovine and rat epidermis. The HCE-2 antibody appears to identify a new class of basic protein precursors of mammalian cornified envelope.

Recurrent hepatitis B, hepatitis C, and combined hepatitis B and C in liver allografts: a comparative pathological study.

Although recurrence of viral hepatitis in liver transplants is common, data comparing recurrent hepatitis B (HB), hepatitis C (HC), and co-existing dual hepatitis B and C (HB&C) are sparse. Posttransplantation liver biopsies, along with molecular, serological, immunohistochemical, and clinical data from 27 patients with pretransplantation diagnosis of chronic viral hepatitis, were reviewed. The patients were placed into 4 groups: Group I, with pretransplantation HB (n = 8); group II, with pretransplantation HC (n = 10); group III with pretransplantation HC and anti-HB surface or core antibody (n = 4); and group IV, with pretransplantation HB&C (n = 5). The histopathologic findings and patient outcome were compared in the 4 groups. A high rate of recurrence of viral hepatitis was seen for all 4 groups: Group I = 100%, group II = 90%, Group III = 100%, and group IV = 80%, with the mean (median) recurrence time of 308 (224), 82 (52), 61 (64), and 125 (70) days, respectively. The number of deaths (their median survival times) were: group I = 4 (374 days), group II = 4 (794 days), group III = 1 (1,143 days), and group IV = 5 (448 days). The earliest histological findings of lobular injury was the presence of acidophil bodies and Kupffer cell hyperplasia, the latter being more prominent in recurrent HC cases. Recurrent HB presented in 2 forms: early (before 150 days) with poor survival and with either severe necroinflammatory histology or with features of fibrosing cholestatic hepatitis, and delayed (after 150 days), with mild necro-inflammatory activity and prolonged survival. HC with or without anti-HB antibodies had early recurrence, but the course was slowly progressive. Patients with HB&C had recurrence of both viruses; however, the course was dictated by HB virus.

Juvenile xanthogranuloma. Clinical and pathologic characterization.

The changing name of the juvenile xanthogranuloma bears witness to the evolution of knowledge and experience of its varied clinical and histologic presentations. This study characterizes the clinical, microscopic, and some immunohistochemical features of 34 cases. The salient clinical findings include a bimodal age distribution inclusive of adults, a male:female ratio of 4:1, occasional multiplicity of lesions (20%), and common presentation in the cephalad area. Histologic findings include varied architectural patterns, cellular participation in various proportions by foamy histiocytes, epithelioid monocytes, lymphocytes, plasma cells, eosinophils, Touton giant cells, and spindle cells of two forms (dendritic and fusiform). S100-positive dendritic cells comprised a minor, but important, component at expansion zones. The significance of these findings is described.

IgG-mediated intravascular hemolysis and nonoliguric acute renal failure complicating discontinuous rifampicin administration.

Acute renal failure following intermittent or discontinuous rifampicin therapy is a relatively infrequent clinical observation. Many pathogenetic mechanisms for the renal failure have been proposed, including intravascular hemolysis with hemoglobinuria and its consequent nephrotoxicity. We report the case of a patient who used rifampicin in a discontinuous fashion and developed hemolysis with nonoliguric acute renal failure. Most reported cases of antirifampicin antibodies are of the IgM class; thus, the development of an IgG antirifampicin antibody is of interest, especially because of its strong in vivo hemolytic properties. In addition, this patient developed a nonoliguric uremic course that did not require dialysis, both of which are distinctly unusual for this clinical setting.

Association between endothelial cell proliferation and pathologic changes in experimental alcoholic liver disease.

We evaluated the role of endothelial cell (EC) proliferation in experimental alcoholic liver disease (ALD) using different dietary models of ALD. Rats were divided into treatment groups to receive ethanol with either corn oil (CO + E), fish oil (FO + E), saturated fat (SF + E), and corn oil with a novel quinone derivative (E3330) and cimetidine. All ethanol-fed rats in the different groups were pair-fed with dextrose replacing the ethanol-derived calories. All ethanol-fed groups and their controls were fed for periods ranging from 1 to 4 weeks. For each animal, immunocytochemical staining for PCNA was performed on paraffin-embedded tissue sections and the number of endothelial cells staining for PCNA per 10 high-power fields (HPF) was determined. Rats fed CO + E and FO + E developed pathological changes, whereas none of the histologic features of ALD were seen in SF + E rats. The severity of injury was reduced in the quinone and cimetidine-treated groups. A higher rate of EC proliferation (6-30x) was seen in the SF + E group (no liver injury) than in the CO + E and FO + E groups (pathologic changes present). The difference was evident by 1 week, which is well before pathologic changes can be recognized (usually 4 weeks). An increase in EC proliferation was seen in the E3330 and cimetidine-treated groups. Our study indicates that the proliferative response of EC in ethanol-fed rats may be a factor in the progression to liver injury. Suppression of ethanol-induced EC proliferation in CO + E and FO + E groups occurred prior to development of liver injury; a lack of suppression of EC proliferation is associated with absence (SF + E and CO + E + cimetidine) or reduction in severity (CO + E + E3330) of liver injury.

Angiogenesis in invasive squamous cell carcinoma of the lip: tumor vascularity is not an indicator of metastatic risk.

BACKGROUND: Recent studies have shown that tumor growth beyond a certain size requires angiogenesis. Microvessel density has, moreover, correlated with metastatic risk in some tumors. Invasive squamous carcinoma (SCC) can develop in the epithelium of the lip and metastasize even when relatively small. This study investigates neovascularization and its relationship to metastatic risk in this tumor. METHODS: All 41 primary SCCs of the lip diagnosed at our institution from 1960 to 1991 were immunostained for factor VIII. Microvessel grade (Mv) from 1+ to 4+ and the average number of vessel profiles (TMvD) in the highest density 200 x (0.785 mm2) and 400 x (0.196 mm2) microscopic fields were determined. TMvDs were compared with those of adjacent non-tumor tissue (NTMvD). Normalized counts (TMvDns) were calculated as TMvD/NTMvD. TMvDs and TMvDns of metastasizing (N = 10) and non-metastasizing (N = 31) tumors were compared (student t-test). RESULTS: In all SCCs TMvDs exceeded NTMvDs (50 vs. 35 at 200 x, P = 0.0014, and 26 vs. 14 at 400 x, P < 0.0001). Metastasizing and non-metastasizing tumors did not, however, differ in Mv, TMvD, or TMvDn. CONCLUSIONS: Angiogenesis develops, but is not quantitatively related to metastatic risk, in primary invasive squamous cell carcinoma of the lip.

Histologic correlates of metastasis in primary invasive squamous cell carcinoma of the lip.

Clinical and histologic features of 44 invasive squamous cell carcinomas of the lip treated by surgical excision were correlated with metastases and survival. Age, diameter, ulceration, grade, depth of invasion, muscle invasion, mitotic rate, lymphatic-vascular invasion, and tumor infiltrating lymphocytes were evaluated. Two (4.5%) patients had local recurrence, 10 (23%) had metastases to cervical lymph nodes, and 2 (4.5%) died from complications of extensive spread in the neck. One patient with local recurrence had regional node metastases. Regional metastases were discovered at the time of (3 patients) or up to 21 (median 8) months after initial diagnosis, correlating with ulceration (P = 0.003), depth of invasion (P = 0.0001), and mitotic rate (P = 0.05) of the primary tumor. Depth exceeding 2.0 mm. was associated with metastasis (P = 0.028), and all carcinomas deeper than 6.0 mm. metastasized. Eight of the 10 patients with regional metastases had no further disease after treatment through the follow-up period (median 64 months). Both fatalities were older (median 81 years) men. These results underscore the metastatic potential of invasive squamous cell carcinoma of the lip. Histologic analysis of the primary lesion can be used to identify individuals at risk.

S100+ cell response to squamous cell carcinoma of the lip: inverse correlation with metastasis.

Previous work has suggested a key role of dendritic cells in antineoplastic immunity. The course of mycosis fungoides and cancers of the lung, colon, thyroid and stomach has been associated with dendritic cell response to the primary tumor. However, this has not been reported for cutaneous or mucosal squamous cell carcinoma (SCC). Thirty-six cases of primary SCC of the lip mucosa or vermillion border, including nine cases with regional metastasis, were studied to investigate the relationship of dendritic cell density with age, tumor grade, mitotic rate, diameter, ulceration, depth of invasion, muscle invasion, tumor-infiltrating lymphocytes (TILs) and metastasis. Dendritic cells were identified using S100 immunohistochemistry, and their peritumor and intratumor density (peri-S100D and intra-S100D) were determined. The mean peri-S100D was 314 +/- 50/mm2. High peri-S100D was associated with lower rate of metastasis (P = 0.03), and no case with peri-S100D > 311/mm2 metastasized. Peri-S100D inversely correlated with depth of invasion (P = 0.04) and ulceration (P = 0.02), and positively associated with TILs (P = 0.02). The mean intra-S100D was 317 +/- 42/mm2. Intra-S100D did not quantitatively correlate with metastasis; however, no metastasis occurred when intra-S100D exceeded 515/mm2. Intra-S100D correlated with brisk TILs (P = 0.04). These results suggest a functional role of dendritic cells in the immune response to SCC. Peri-S100D may be a prognostic indicator.

Microscopic localization of calcifications in and around breast carcinoma: a cautionary note for needle core biopsies.

OBJECTIVE: To detail the microanatomic localization of microcalcifications (Ca++) occurring in association with breast carcinoma and thereby to determine their reliability as a marker of breast carcinoma in small tissue core biopsies. SUMMARY BACKGROUND DATA: Identification of the pathology associated with Ca++ in mammograms has acquired increasing importance in the early detection of breast carcinoma. With recent advances enabling computer-guided stereoscopic needle biopsy of calcified foci, histopathologic diagnosis is rendered on increasingly small tissue samples, raising the risk of misdiagnosis. Knowledge of the microanatomic distribution of Ca++ in relation to diagnostic epithelial elements is essential for assessing their significance in small tissue biopsies. METHODS: All 32 carcinomas with Ca++ within 1 cm of carcinoma diagnosed by open biopsy at the New England Deaconess Hospital from January 1994 to January 1995 were studied. Ca++ were classified as being within ductal or lobular carcinoma in situ, invasive carcinoma, carcinoma-associated stroma, benign stroma >1 mm from carcinoma, or benign ducts or terminal duct-lobular units. If Ca++ were peritumoral, their distance from the tumor was measured. RESULTS: Ca++ were present only in malignant components in 31%, only in benign components in 34%, and in both in 34% of cases. The most common locations of Ca++ were benign peritumoral ducts (62%) and ductal carcinoma in situ (54%). The microanatomic distribution of benign peritumoral Ca++ in relation to the mass is detailed. CONCLUSIONS: In carcinomas with Ca++ in the area of tumor, Ca++ may not be localized to malignant tissue. Caution should be used when interpreting the finding of Ca++ in benign components of small tissue samples of breast masses.

Cutaneous hamartoma of adnexa and mesenchyme. A variant of folliculosebaceous cystic hamartoma with vascular-mesenchymal overgrowth.

We describe a hamartoma with folliculosebaceous and mesenchymal components. Striking vascular-mesenchymal elements distinguish this entity from previously described hamartomas and other adnexal proliferations. It is also distinct from the recently described folliculosebaceous cystic hamartoma but likely represents a variant with mesenchymal overgrowth.

Chronic varicella zoster infection mimicking a basal cell carcinoma in an AIDS patient.

Chronic herpesvirus infections are common in patients infected with HIV. Atypical skin lesions secondary to long-standing varicella-zoster virus (VZV) infection have been reported. We present a case of an AIDS patient with a chronic VZV infection that simulated a basal cell carcinoma. Histologic examination and immunohistochemistry confirmed the presence of the virus in the follicular epithelium. In the immunocompromised patient, biopsies should be performed on all suspicious lesions because medically-treatable infections may take on the appearance of malignancy.

Stromal tumors of the stomach: prognostic value of the PCNA index.

Reliable indicators of behavior in stromal tumors of the gastrointestinal tract have yet to be elucidated. Aggressive behavior has been associated with large size, hypercellularity, tumor necrosis, nuclear atypia, and high mitotic rate. Recently, new methods of measuring proliferation have been developed that exploit the phenomenon of cell cycle specific protein synthesis. In this study the expression of the S-phase specific nuclear protein proliferating cell nuclear antigen (PCNA) is tested as an indicator of malignancy. Sixteen stromal tumors of the stomach were reviewed for tumor size, cellularity, nuclear atypia, mitotic rate, necrosis, vascular invasion, and predominant cell type. Local recurrence and/or mortality were ascertained with a minimum follow-up of 5 yr. An immunohistochemical assay for PCNA was performed on a paraffin section of tumor and the percentage of positively stained cells ("PCNA index") was determined. Among the nine men and seven women, age 37 to 80 (median 66) yr, two had local recurrences that were treated surgically with no metastases. Only positive resection margin correlated with local recurrence. Two other patients developed distant metastases at 8 and 15 mo and died. Mortality did not correlate with age, sex, size, cellularity, necrosis, cell type, and vascular invasion. High grade nuclear atypia (2/2 versus 1/14, P = 0.02) and high mitotic rate (20.5 versus 5.5 per 40 hpfs, P = 0.01) did correlate with mortality. PCNA index did not correlate with local recurrence, but was sharply higher in fatal cases (6.4 versus 1.2, P = 0.001). Both fatal tumors had PCNA values above 6.0, and all others had values of 4.0 or less. The PCNA index is a proliferative marker that may have prognostic value in gastric stromal tumors.

Nucleolar organizer regions (AgNOR) and Ki-67 immunoreactivity in cutaneous melanocytic lesions.

A stepwise schema for the developmental biology of malignant melanomas has been proposed. In this study, two properties associated with malignant transformation--AgNOR production and Ki-67 immunoreactivity--were tested on a series of lesions representative of this model. Fourteen ordinary nevi (N), seven dysplastic nevi (DN), 12 radial growth phase melanomas (RGPM), and eight vertical growth phase melanomas (VGPM) were stained for AgNORs; Ki-67 immunoreactivity was determined in paraffin sections after antigen retrieval. AgNORs averaged 2.1 in N, 3.8 in DN, 5.4 in RGPM, and 7.3 in VGPM (p < 0.001 between all groups). Ki-67 immunoreactivity averaged < 1 in nevi, 1.6 in dysplastic nevi, 23.3 in RGPM, and 20.1 in VGPM. No difference was observed between N and DN, and RGPM/VGPM had higher Ki-67 indices than N/DN (p < 0.01). Stepwise increases in cellular AgNORs parallel the melanocytic progression model and thereby corroborate the intermediate nature of the dysplastic nevus. The order of magnitude increase in Ki-67 immunoreactivity observed in RGP and VGP melanomas compared with N and DN likely reflects the onset of loss of cell cycle control in these lesions.

Paget's disease of the nipple and angiosarcoma of the breast following excision and radiation therapy for carcinoma of the breast.

We report a case of bilateral angiosarcomas developing in breasts after radiation therapy. The angiosarcomas developed 7 years after the first dose and 3 years after the last dose of radiation. In addition, Paget's disease of the nipple was diagnosed in the right breast. c-neu overexpression was noted in the adenocarcinoma but not in the angiosarcoma. Neither tumor was immunoreactive for c-k-ras. The oncogenic expression of the radiation-induced angiosarcoma was different from that of the radiation-resistant adenocarcinoma. The simultaneous occurrence of angiosarcoma and Paget's disease of the nipple has not been reported previously. The importance of recognizing Paget's disease in post-irradiated breasts and complications of breast conservation therapy are stressed.

Protothecosis: an unusual cause of chronic subcutaneous and soft tissue infection.

Protothecosis of subcutaneous and soft tissues is a rare occurrence in humans. We present two patients with chronic subcutaneous protothecosis affecting the elbow and foot respectively. Both patients had been treated with local corticosteroid injections and had recent exposure to water. The diagnosis was made histopathologically in both cases and confirmed by culture in one case. Histopathology showed typical Protothecal sporangia with surrounding mixed inflammatory infiltrate including necrotizing granulomas. Organisms stained positively with periodic acid-schiff, Gomori's methenamine silver, and Gridley fungus stains. In one case, intravenous chemotherapy was required to eliminate the pathogens. Histopathologic identification of the organisms is vital to ensure adequate therapy and avoid chronic smoldering infection.

Atypical angiomatosis in polyarteritis nodosa.

Biopsies from an area of livedo reticularis and adjacent to a leg ulcer in a woman with polyarteritis nodosa showed florid angioendothelial proliferation simulating angiosarcoma. This angioproliferative reaction has not been described previously in polyarteritis nodosa. Its microscopic differentiation from angiosarcoma is important.

Neuromuscular and vascular hamartoma of the small bowel.

Neurovascular and muscular hamartoma is an unusual benign neoplasma of the small intestine. The clinical and pathological features of this lesion, which we recently encountered in a 91-year-old male, are the subject of this report and are discussed in the context of previously described cases.