An immunohistochemical study of p53 protein expression in classical Kaposi's sarcoma.

The present study was performed to determine the frequency of p53 protein immunoreactivity in classical Kaposi's sarcoma (KS) as a whole and in relation to the histological subtypes which are considered to correspond to the developmental stages of the tumor. The accumulation of p53 protein was studied immunohistochemically using monoclonal antibody BP53-12 on formalin-fixed paraffin-embedded sections of 36 KS lesions, of which 14 were classified histologically as early type and 22 as spindle-cell or mixed type. No positive immunoreactivity was detected in any of the 14 early-type lesions. Among the 22 spindle-cell and mixed variants, positive staining was detected in 5-10% of the tumor cells in one lesion, 1-5% of the cells in six lesions, and in < 1% of the cells in two lesions. These very small percentages of positively stained cells in less than half of the cases of the spindle-cell and mixed variants do not support a significant role for p53 in tumor progression and evolution in KS.

The pattern of HMB-45 antibody staining in compound Spitz nevi.

We studied the staining pattern of HMB-45 antibody in 29 compound Spitz nevi (SNs) of the epithelioid cell variety, 17 of which showed extension of nevus cells into the reticular dermis (i.e., "deep"); 20 ordinary compound nevi (CNs), all with a deep dermal component; and 22 primary cutaneous invasive malignant melanomas (MMs) (excluding the desmoplastic and spindle cell types), 12 of which extended into Clark level IV or V. Of the 29 SNs, eight (28%) stained negatively; five (17%), including two deep SNs, stained in the epidermal component only; and 16 (55%), including 10 deep SNs, stained in both the epidermal and dermal components. Of the latter 10 deep SNs, eight stained in the upper dermis only, and in the remaining two lesions, a smaller number of positively stained nevus cells were detectable in the lower dermis as well; these two SNs were not atypical histologically. Of the 20 CNs, four (20%) stained negatively, two (10%) stained in the epidermal component only, and 14 (70%) stained in the epidermal component and the upper dermis only. Of the 22 MMs, one stained negatively, and 21 (95%) stained positively in both the epidermal and dermal components. The pattern was variable in frequency of both staining and distribution, but showed no stratification. We conclude that the majority of our positively stained deep compound SNs showed a stratified pattern of HMB-45 staining, similar to ordinary CNs and different from MMs, and that this pattern might be used as an adjunct in the histopathologic differential diagnosis of compound SN and MM, in the proper clinicopathological context.

Immunohistochemical study of p53 protein expression in Spitz nevus as compared with other melanocytic lesions.

The accumulation of p53 protein was studied immunohistochemically on paraffin-embedded sections of 26 Spitz nevi (SNs), 26 primary invasive cutaneous malignant melanomas (MMs), 20 metastases of MM, and 17 ordinary compound nevi (CNs), using monoclonal antibody BP53-12. Positive reactivity was detected in some of the tumor cells in seven (35%) metastatic MMs, all exhibiting strong nuclear staining; eight (31%) primary MMs, of which seven showed strong nuclear staining; two (7%) SNs, of which only one showed strong nuclear staining; and none of the CNs. The frequencies of the positively stained lesions in general, and the strongly positively stained lesions in particular, in the MM and metastatic MM groups were each statistically significantly higher than the respective frequencies in the SN and CN groups. We believe that the immunohistochemical detection of p53 protein with the use of monoclonal antibodies such as BP53-12 on paraffin sections, especially when strong nuclear reactivity is demonstrated, may prove to be an adjunctive tool in the histopathologic differentiation of MM from SN.

Histopathologic findings in the clinically uninvolved skin of patients with mycosis fungoides.

Several of the methods of treating plaque-stage mycosis fungoides (MF) advocate treatment of the whole skin, although relatively little information exists on the histologic appearance of clinically uninvolved MF skin. Our study was performed to elucidate the histologic appearance of the clinically uninvolved skin of plaque-stage MF patients. Biopsies were taken from the clinically uninvolved skin of 18 untreated patients with plaque-stage MF, 1 cm (near) and > or = 10 cm (far) from the MF plaques, and from 23 normal, healthy controls. Eight to 12 serial sections from each specimen were examined. The most common histologic finding, which was observed in six (33%) biopsies of near skin and four (22%) biopsies of far skin compared with only one (4%) biopsy of normal control (p = 0.02 and 0.1, respectively), consisted of mononuclear-cell infiltrate around a blood vessel in the papillary dermis, with extension of some of these cells into the overlying epidermis. More diffuse and epidermotropic mononuclear cell infiltrates were seen in an additional three (17%) biopsies of near skin, whereas mild nonepidermotropic superficial perivascular or perifollicular mononuclear cell infiltrates were observed in an additional four (17%) biopsies of the normal controls. In conclusion, the most common histologic finding in our study, which was observed predominantly in the MF group, might represent an earlier stage and clinically undetectable involvement of the normal-looking skin in MF patients.

An immunohistochemical study of estrogen and progesterone receptors in adenocarcinoma of the endometrium and in the adjacent mucosa.

The immunoperoxidase stain for estrogen and progesterone receptor content in endometrial adenocarcinoma was correlated with the grade and stage, level of myometrial invasion, age and survival of the patients. Anti-estrogen and anti-progesteone receptor monoclonal antibodies were applied to paraffin-embedded tissue from hysterectomy specimens of 100 patients with adenocarcinoma of the endometrium. In 34 of the cases the receptors were studied in the endometrium adjacent to the tumor and compared to the nuclear receptor content in the carcinoma. There was a high inverse correlation between the estrogen receptor status and the grade of tumor (R = - 0.45, P = 0.006). The estrogen receptor measured in the endometrium near the tumor showed a negative correlation with the grade of the tumor (R = -0.42, P = 0.013). The estrogen, but not the progesterone, receptor content, was positively related to the age of the patient (P < 0.05). No significant correlation of the receptor status with the depth of myometrial invasion was found, despite the obvious interdependence between the grade and myometrial invasion. The progesterone receptor staining index appeared to be a distinct independent prognostic factor in endometrial cancer. The immunohistochemical analysis of the steroid hormone status in endometrial cancer therefore offers an alternative to the quantitative ligand-binding assay.

Mast cells in colorectal neoplasias and premalignant disorders.

PURPOSE: Mast cells have recently been found to be well correlated with the prognosis of patients with rectal cancer. This work aimed to characterize the role of mast cells in colonic premalignant conditions. METHODS: Mast cells were quantified in various colonic disorders, particularly those with premalignant potential. Possible avenues of mast cell action were investigated using these tissue samples, by measuring basement membrane and collagen layer thickening. RESULTS: The mean number of mast cells in carcinoma sections was 0.967/0.9 mm2, in various colorectal neoplasias and related conditions it ranged from 1.36-3/0.9 mm2, and in normal histologic specimens it was 11.90/0.9 mm2. These data established statistically significant differences in mast cell numbers in the colonic disorders studied. The number of mast cells is greatest in the lamina propria level of the colon, a site often not examined because of the limited depth of samples obtained from endoscopic biopsies. CONCLUSIONS: Mast cell numbers were found to be correlated to the development from premalignancy to colonic malignancy. Mast cells may be useful as markers of colorectal neoplasia.

A comparative study of radioligand (DCC) and modified immunoperoxidase anti-estrogen receptor techniques in breast carcinoma.

This study deals with the pattern of estrogen receptors in 52 cases of invasive breast carcinoma by comparing the ligand-binding assays with dextran-coated charcoal (DCC) to an immunoperoxidase assay (IPE) technique in paraffin-embedded material from the same tumor. A modification of the IPE by adding cobalt chloride in the final reaction and counter-staining with eosin instead of hematoxylin was introduced. A significantly high correlation was found between the two compared methods (80.8% positivity by the DCC method and 80% by the IPE method). The correlation was very significant in cases where the immunohistochemical grading was higher than 4 (according to our semi-quantitative evaluation scale of 0-8), and for these cases we suggest the IPE method as highly specific and precise.

Epithelial markers in malignant melanoma. A study of primary lesions and their metastases.

In order to determine epithelial markers in malignant melanoma in routinely processed paraffin sections and to compare the staining of primary (cutaneous) malignant melanomas and their metastases, we stained formalin-fixed paraffin sections of 13 primary and 18 metastatic malignant melanomas using the streptavidin-biotin peroxidase method by antibodies to S-100, vimentin, HMB-45, polyclonal carcinoembryonic antigen (CEA), monoclonal CEA, cytokeratins (CAM 5.2 and broad-spectrum CKKES), and epithelial membrane antigen (EMA). All primary and most metastatic malignant melanomas showed positive staining with anti-S-100, HMB-45, and anti-vimentin. Reactivity with polyclonal CEA was observed in 15 (48%) of the 31 lesions; 14 of them were metastatic. No lesion was reactive with monoclonal CEA. Significant cytokeratin (CK) staining was evident in only three (9.7%) lesions (all metastatic), which also stained specifically with anti-CK 18. EMA was observed only focally in two (6.5%) lesions. There was no correlation between epithelial markers staining of the primary tumours and their metastases. All lesions with CK or EMA staining showed concomitant extensive staining for S-100, HMB-45, and vimentin. We conclude that (a) polyclonal CEA staining in malignant melanoma is not rare and is probably due to CEA-related molecules; (b) significant CK reactivity is rare and related to simple CK, such as CK 18; (c) epithelial marker reactivity is more common in metastases of malignant melanomas and is not correlated to the reactivity in their primary tumors. Considering our results and reports of positive S-100, vimentin, and HMB-45 in epithelial tumors, a wide panel of antibodies is recommended for the study of undifferentiated tumors.

Classic and iatrogenic Kaposi's sarcoma. Histopathological patterns as related to clinical course.

The histological findings in the first diagnostic biopsy of 50 patients with Kaposi's sarcoma (KS) were classified and were related to the clinical course. Forty-one patients had the "classic KS," of whom 31 had a benign course and 10 an aggressive disease. Nine patients had iatrogenically induced KS. Histologically the following subtypes were found: mixed type in 18 patients, spindle type in 16, early type in 14, and hemangiomatous and lymphangiomatous types in one each. We did not find any statistical relationship between the histological subtypes, degree of nuclear atypia, number of mitoses, and density of the inflammatory cell infiltrates in the KS lesions, and the clinical types and course of the disease. The results of this study do not support the possibility of a prognostic importance regarding the histological features of the early lesions in patients with classic or iatrogenic Kaposi's sarcoma.

Localized granuloma annulare. Histopathological and direct immunofluorescence study of early lesions, and the adjacent normal-looking skin of actively spreading lesions.

We studied 10 biopsies of early (< 1 week) localized granuloma annulare (LGA) lesions and 10 specimens of the normal-looking skin adjacent to actively spreading LGA lesions for signs of vasculitis, using histological and direct immunofluorescent (DIF) techniques. In the early LGA lesions, the collagen fibers showed various forms of alterations, with hyalinization and fragmentation being the most common; in half of these lesions, neutrophils and nuclear fragments in various numbers and densities were found among the altered collagen fibers. Some of the small blood vessels in the areas of granulomatous inflammation showed endothelial cell hypertrophy in four cases, and in one case also endothelial cell proliferation and luminal occlusion. In only one case, a single small blood vessel, which was situated in the center of a palisading granuloma, showed fibrinoid necrosis of its walls. The DIF study of all 20 specimens and the histological study of the 10 specimens of normal-looking adjacent skin did not reveal immune deposits in the vessels' walls, or histological evidence of vasculitis, respectively. We believe that these findings do not support a role of vasculitis in the formation of LGA lesions, but suggest that neutrophils might play a primary or secondary role in their pathogenesis.

Antibiotic treatment of experimental Pseudomonas aeruginosa pneumonia in guinea pigs: comparison of aerosol and systemic administration.

Pseudomonas aeruginosa pneumonia is associated with high mortality despite appropriate systemic antibacterial therapy. To examine the effects of aerosolized tobramycin on experimental P. aeruginosa pneumonia, guinea pigs were assigned to two main groups: a study group, in which P. aeruginosa was instilled into the tracheal lumen, and a control group, in which normal saline was instilled. Each group was then subdivided according to treatment: normal saline or tobramycin by intramuscular injection, normal saline or tobramycin by aerosol, or normal saline or tobramycin via both aerosol and intramuscular injection. Compared with either intramuscular or aerosolized tobramycin, the combination of aerosol plus intramuscular tobramycin achieved slightly higher survival, total eradication of P. aeruginosa from lungs, and a significant further reduction of lung inflammatory changes. Adding aerosolized tobramycin to systemic tobramycin in the treatment of experimental P. aeruginosa pneumonia enhanced eradication of P. aeruginosa and mitigated pulmonary inflammatory changes.

Muscle involvement in Walker-Warburg syndrome. Clinicopathologic features of four cases.

Walker-Warburg syndrome is a lethal autosomal recessive disorder characterized by anomalies of the central nervous system and the eye. Recently, elevation of muscle enzymes and congenital muscular dystrophy have been found to contribute to the diagnosis of this syndrome. The authors studied two pairs of siblings with Walker-Warburg syndrome, offspring of consanguineous parents from two unrelated families. The patients had hydrocephalus, congenital glaucoma, elevated muscle enzymes, and hypotonicity. The histologic and ultrastructural muscular changes were consistent with congenital muscular dystrophy.

CEA reactivity in amelanotic malignant melanoma of the anal canal.

Amelanotic malignant melanoma can present a diagnostic problem in histopathology, especially when it presents in an extracutaneous location. In such cases electron microscopy and/or immunohistochemistry are invaluable for diagnosis. A 63-year-old women with rectal bleeding was found to have an amelanotic malignant melanoma of the anal canal, with compatible clinical and gross pathology and light and electron microscopic findings. Tumor cells showed positive staining with antibodies to vimentin, S 100 protein, and HMB-45 (melanoma-specific antibody), but also with polyclonal antibody to carcinoembryonic antigen (CEA). Tumor cells failed to stain with monoclonal antibody to CEA. The nature and significance of CEA reactivity in malignant melanoma are discussed.

Dermatofibrosarcoma protuberans appearing during pregnancy.

Dermatofibrosarcoma protuberans is an uncommon skin tumor with a high potential for local recurrence if not adequately excised. There are only two reports in the literature describing dermatofibrosarcoma protuberans that enlarged considerably during pregnancy. On the other hand, the new development or enlargement of dermatofibromas in pregnancy is well documented. We present 2 additional patients in whom a dermatofibrosarcoma protuberans appeared and grew rapidly during pregnancy. Immunohistochemical studies were negative for estrogen and progesterone receptors. The possible pathophysiology and surgical management are presented and discussed.

Müllerian adenosarcoma presenting as cervical polyps: a report of seven cases and review of the literature.

OBJECTIVE: To emphasize the importance of early diagnosis in cases of müllerian adenosarcoma that appeared as benign-looking cervical polyps. METHODS: We examined seven cases of müllerian adenosarcoma of the uterus in patients 14-63 years of age (median 39 years). Tissue protruding from the external os and an initial diagnosis of a cervical polyp were common findings for all patients. On repeated examination, all lesions were interpreted as müllerian adenosarcomas. RESULTS: Histologic examination demonstrated benign glands with a sarcomatous stroma, which typically formed periglandular cuffs of increased cellularity. The sarcomatous stroma was homologous in four cases and contained heterologous elements such as striated muscle, lipoblast, and cartilage in three cases; one patient had a sarcomatous overgrowth of stromal elements. The question of a müllerian adenofibroma versus adenosarcoma was raised in three cases with the general appearance of slit-like glands surrounded by a stroma with fibrosis and a low mitotic rate. Using the criteria of stromal cellularity--marked stromal atypia and a mitotic index of two figures per ten high-power fields--the cases were classified as adenosarcomas. The sarcomatous overgrowth, the presence of heterologous elements, and a high mitotic rate seem to be important prognostically. CONCLUSION: Gynecologists and pathologists should be aware of the difficulties and delay in the diagnosis of müllerian adenosarcoma when the tumor presents as a benign-looking cervical polyp.

Immunohistochemical staining techniques of intraocular tumors.

The peroxidase-antiperoxidase staining technique for the antigens S-100, neuron-specific enolase, and alpha 1-antichymotrypsin (alpha 1ACT) was applied to 57 intraocular tumors: 46 malignant melanomas of the uvea, seven retinoblastomas, and four tumors metastasizing to the eye. The staining characteristics of the different intraocular tumors were compared. Staining for S-100 in a fine-needle aspiration biopsy sample taken from a malignant melanoma of the choroid before enucleation of the globe was attempted. The positive staining of a few cells thus obtained suggested that this technique may be helpful in the diagnosis of melanomas. The alpha 1ACT stain used in this study has not been used previously in ophthalmology to our knowledge. We found 60% of malignant melanomas of the choroid stained positively. Another finding was the staining of the retinal pigment epithelium with alpha 1ACT in 30% of eyes with malignant melanoma of the uvea.

Porokeratotic eccrine ostial and dermal duct nevus. An abnormally keratinizing epidermal invagination or a dilated, porokeratotically plugged acrosyringium and dermal duct?

Porokeratotic eccrine ostial and dermal duct nevus (PEODDN) has been said to represent a widely dilated, keratin-plugged acrosyringium and dermal duct. We have observed in a case of congenital PEODDN a normal-appearing, acrosyringium-like duct that traverses vertically the entire length of the parakeratotic column. Also, in its lower course, it stained positively for carcinoembryonic antigen, while the inner borders of the invagination from which the parakeratotic column arose stained negatively. This leads us to suggest that the epithelial structure in PEODDN is an abnormally keratinizing epidermal invagination through which an acrosyringium-like duct traverses, rather than an abnormally dilated, parakeratotically plugged acrosyringium and dermal duct.

Signet-ring cell carcinoma of the prostate mimicking primary gastric carcinoma.

A post mortem examination of a 70 year old man, who died three years after a poorly differentiated adenocarcinoma of prostate had been diagnosed, showed widespread signet-ring cell carcinoma, with an associated linitis plastica. The signet-ring cells stained positively with prostatic specific antigen and with prostatic specific acid phosphatase, but failed to react with mucopolysaccharide staining. The electron microscopic appearance of the signet-ring cell tumour was due to the presence of large cytoplasmic vacuoles. This case emphasises the possibility that cases of metastatic signet-ring cell carcinoma may be prostatic in origin. This can be confirmed by specific immunohistochemical studies.