Three-dimensional evaluation of subclinical extension of extramammary Paget disease: visualization of the histological border and its comparison to the clinical border.

BACKGROUND: In extramammary Paget disease (EMPD), Paget cells are sometimes detected outside the clinical border (subclinical extension). However, the spreading pattern of Paget cells in subclinical extension remains unclear. In addition, the macroscopic appearances of lesions accompanied by subclinical extension are totally unknown. OBJECTIVES: To characterize the spreading pattern of Paget cells as well as the macroscopic appearance of lesions of EMPD with subclinical extension. METHODS: Nineteen patients with primary anogenital EMPD underwent mapping biopsies and excisional surgeries; biopsy samples were then taken at the periphery of well-demarcated lesions. Samples were transparentized and subjected to whole-mount immunostaining with anticytokeratin 7 antibody to label Paget cells. The histological border was evaluated in three dimensions by two-photon microscopy. The shape and location of the histological border were compared with those of the clinical border. RESULTS: In 21 samples taken at the lesion where subclinical extension was not shown by mapping biopsy, the shape and location of the histological border were almost identical to those of the clinical border. However, two samples exhibited small foci of Paget cells outside the clinical border, showing subclinically extended satellite lesions. In the two samples taken at the lesions where subclinical extension was shown by mapping biopsy, a continuous arrangement of Paget cells extending beyond the clinical border was identified. Subclinically extended Paget cells were detected solely outside hypopigmented patches with erythema. CONCLUSIONS: In EMPD, at least two patterns of subclinical extension exist: continuous and satellite lesions. Subclinical extension might exist preferentially outside hypopigmented patches with erythema.

Perturbations of both nonregulatory and regulatory FOXP3+ T cells in patients with malignant melanoma.

BACKGROUND: 'FOXP3+ regulatory T cells' (Tregs) are reported to be increased in tumour-bearing hosts including patients with melanoma, leading to tumour immune suppression. However, this idea is challenged by recent evidence that the 'FOXP3+ Treg' fraction in fact contains activated 'nonregulatory' T cells. Also, FOXP3+ T cells are reported to have functionally and kinetically distinct subsets. OBJECTIVES: To investigate whether either or both of regulatory and 'nonregulatory' FOXP3+ T cells are perturbed in patients with melanoma. METHODS: FOXP3+ T cells were classified into three subsets, namely CD45RO+FOXP3(low) nonregulatory T cells, CD45RO+FOXP3(high) effector Tregs, and CD45RO-FOXP3(low) naïve Tregs, according to their expression levels of FOXP3 and CD45RO. The percentage and cytokine production of these FOXP3+ T-cell subsets were assessed by flow cytometry. RESULTS: Both regulatory and nonregulatory T cells were increased in patients with melanoma. Moreover, we found three unexpected perturbations in FOXP3+ T-cell subsets: (i) patients with melanoma showed higher frequencies of FOXP3(low) nonregulatory T cells, which decreased and normalized after tumour removal; (ii) FOXP3(low) naïve Tregs containing higher frequencies of interferon-γ+ cells increased with tumour progression; and (iii) CD45RO+FOXP3(high) effector Tregs were pronouncedly infiltrated around tumour tissues. CONCLUSIONS: These findings demonstrate that patients with melanoma have distinct and differential perturbation of both regulatory and nonregulatory FOXP3+ T cells. The degree of perturbation is associated with tumour burden and progression, suggesting that the perturbation reflects fundamental pathophysiological processes in patients with melanoma. The presented analysis provides a practical approach to investigate the immunological environment of cancer patients.

Churg-Strauss syndrome with renal involvement: a case report.

We report a case of Churg-Strauss syndrome with renal involvement. A 51-year-old woman was admitted to our hospital in April of 1999 complaining of abdominal pain and severe diarrhea followed by multiple purpura on both legs. A diagnosis of Churg-Strauss syndrome was made on the basis of the presence of preceding asthma, eosinophilia, increased IgE, and the histological findings of a biopsy taken from the purpura. Creatinine clearance level at admission was low (43.3 ml/min) and urinalysis revealed proteinuria and microscopic hematuria. These abnormalities were improved with systemic steroid therapy. Renal involvement seems to be more common than previously reported in Churg-Strauss syndrome.

[Infratentorial oligodendroglioma: report of a case].

Oligodendroglioma occurs primarily in the cerebral hemisphere in adult. A rare case of oligodendroglioma in the cerebellum is presented, and previous reported cases were reviewed. A thirty-one year old female was admitted to our service only with headache. CT scan demonstrated a low density mass in the vermis of the cerebellum which was not enhanced. Bilateral vertebral angiography showed an avascular mass in the vermis. The preoperative diagnosis was astrocytoma or ependymoma, and a suboccipital craniotomy was performed. The tumor which was soft, yellowish gray and well-circumscribed, was developed from the vermis and extended into the cisterna magna. The tumor which size was approximately 3 X 4 X 5 cm. Microscopically the specimen showed round, darkly stained nuclei and clear perinuclear cytoplasmic halos. Moreover, the immunoperoxidase method testified the absence of glial fibrillary acidic protein in the tumor cells. The pathological diagnosis was oligodendroglioma. Post-operatively the patient was doing well without any complications. There was no clinical nor CT evidence of tumor recurrence forty months after resection. The 11 reported cases of infratentorial oligodendroglioma including ours were analyzed and the following conclusion was obtained. Infratentorial oligodendroglioma occurs in the younger age group. The tumor has special tendency to form cyst. Frequency of calcification is low. Prognosis is good if the tumor is resected in early stage. Pre-operatively, however, it was difficult to differentiate oligodendroglioma from astrocytoma.

[A case of malignant melanoma with orbital metastasis which caused the first symptoms].

A thirty-four-year-old man was admitted to our hospital because of the disturbed visual acuity and pain on the eye movement of the right eye. He had prominent right eye and CT-scan and MRI of the brain disclosed a tumor which could be obviously distinguished from the extraocular muscles, optic nerve and the bulb of eye in the retrobulbar region. On operation we identified dark-red solid tumor which was 3.0cm in diameter, and diagnosed it malignant melanoma pathologically. Because postoperative study detected amelanotic melanoma in the white patch on the right upper extremity, this right orbital tumor was considered to be the metastasis of it from the right upper extremity. Metastatic malignant melanoma of the skin to the orbit is very rare, while most of the eye-associated malignant melanoma originates from uveal tract, special choroid, and conjunctiva. This case was the 26th case of these in the world and the first case in Japan, furthermore the 4th case in the world whose first symptoms were caused by the orbital metastasis.

Isolation and partial characterization of eosinophil granule proteins in rats--eosinophil cationic protein and major basic protein.

Peritoneal eosinophilia was induced in rats using Ascaris suum extract as an antigen, and characteristics of granule proteins of eosinophils collected from the peritoneal cavity were investigated. Peritoneal eosinophilia was induced by injection of the antigen solution into the peritoneal cavity of the immunized rats that had been orally administered with cyclophosphamide. Peritoneal cells were collected 48 h after injection of the antigen solution, incubated in plastic dishes, and nonadherent cells were collected as an eosinophil-rich fraction, from which granule proteins were extracted. Granule proteins were then purified by cation exchange chromatography, gel filtration, copper chelate affinity chromatography, and reverse-phase HPLC. Two distinct basic proteins of which molecular weights are 18- and 17-kD were obtained. Partial N-terminal amino acid sequence analysis revealed that the 18-kD protein and the 17-kD protein were homologous to the human eosinophil cationic protein (ECP) and the human and guinea pig major basic protein (MBP), respectively. Both the two proteins showed strong bactericidal activity against Escherichia coli and Staphylococcus aureus. These results indicate that rat eosinophils also possess ECP and MBP in their granules as well as human and guinea pig eosinophils.

Increase of eosinophilic cell population in bone marrow of rats by immunization with Ascaris suum antigen.

Immunization of rats with the antigen, Ascaris suum extract, increased the number of peripheral eosinophils. Analysis by Western blot and reverse transcription-polymerase chain reaction revealed that the levels of major basic protein and its mRNA in the bone marrow were also increased, suggesting that eosinophilic cell population in the bone marrow is increased by the immunization. These findings indicate that immunization with this antigen stimulates differentiation of progenitor cells to eosinophils in the bone marrow, and induces blood eosinophilia.