Metallothionein expression in basaloid proliferations overlying dermatofibromas and in basal cell carcinomas.

Basaloid proliferations overlying dermatofibromas which morphologically resemble superficial basal cell carcinomas have been interpreted as both reactive/regressive and frankly malignant. Metallothioneins (MTs) are low-molecular-weight proteins with a selective binding affinity for heavy metal ions. MTs has been proposed to represent a biological marker of carcinogenesis and, in a variety of human tumours, a correlation between immunohistochemically overexpression of MT and aggressive clinical behaviour has been shown. In order to clarify the nature of basaloid proliferations overlying dermatofibromas, we examined, immunohistochemically, 10 dermatofibromas with overlying simple hyperplasia, 16 dermatofibromas with overlying basaloid proliferation, and 35 basal cell carcinomas, for expression of MT. In normal epidermis, the basal keratinocytes showed cytoplasmatic MT immunoreactivity. The staining intensity was stronger in the basal cells of the rete ridges, an observation which is in accordance with the high proportion of S-phase cells in this area. Simple hyperplasia showed the same MT expression pattern as normal epidermis. Basaloid proliferations stained like superficial and nodular basal cell carcinomas. Of nodular basal cell carcinomas, 92% (12 of 13) showed decreased/absent MT immunoreactivity, while 86% (six of seven) of infiltrating/morphoea-like basal cell carcinomas showed overexpression of MT (P = 0.001, Fisher's exact test). The results demonstrate that MT overexpression in basal cell carcinomas is correlated with infiltrative growth pattern. The similar expression of MT in basaloid proliferations and 'non-infiltrating' basal cell carcinomas suggests that these lesions share a common change in metabolism and/or differentiation.

Bcl-2 overexpression in basaloid proliferations overlying dermatofibromas and basal cell carcinomas.

Basaloid proliferations overlying dermatofibromas resembling superficial basal cell carcinomas have been interpreted both as reactive/regressive and frankly malignant. Basal cell carcinoma is a slow-growing tumour, which so far has been regarded as an actively proliferating lesion with a high apoptotic activity. We examined immunohistochemically 6,dermatofibromas with overlying simple hyperplasia, 12 dermatofibromas with overlying basaloid proliferations, and 24 basal cell carcinomas for expression of Ki-67 protein, and bcl-2 protein. The Ki-67 labelling index represents an estimate of proliferative activity. Bcl-2 protein suppresses apoptosis. The Ki-67 labelling indexes of basaloid proliferations, basal cell carcinomas, and normal epidermis were similar (11-15%, p < 0.05, Mann-Whitney test). Bcl-2 protein was expressed in all cells of basaloid proliferations, similar to the expression pattern in basal cell carcinomas. We suggest that basaloid proliferations overlying dermatofibromas might have achieved a phenotype that equals an early stage of BCC carcinogenesis.

Correlation of growth fraction by Ki-67 and proliferating cell nuclear antigen (PCNA) immunohistochemistry with histopathological parameters and prognosis in primary breast carcinomas.

The immunoreactivity for Ki-67 and PCNA was investigated in 487 patients with primary breast carcinomas using MIB-1 (Immunotech, France) and PC-10 (DAKO, Denmark) as primary antibodies. Formalin-fixed and paraffin-embedded tissue sections were used. The immunoreactivity for Ki-67 and PCNA was found to be independent of the length of fixation if the sections were pretreated in a microwave oven in citrate buffer and distilled water, respectively. The immunostaining was evaluated semiquantitatively. High Ki-67 score (more than 1% positive tumour cells) and PCNA over-expression (more than 25% positive tumour cells) were correlated with clinicopathological parameters such as large tumour size, high histological grade (poor differentiation), and absence of steroid hormone receptors, which are parameters of an aggressive phenotype of the tumour. In univariate analysis of survival data, both Ki-67 and PCNA were parameters of a poor overall survival in both lymph node-positive and -negative patients. In multivariate anlaysis using a Cox model stratified by nodal status, Ki-67 and PCNA failed to be of prognostic significance whereas classical histopathological parameters such as tumour size and histological grade turned out to be of independent prognostic significance in both lymph node-positive and -negative patients, while progesterone receptors were of independent prognostic significance only in lymph node-positive patients.

Metallothionein expression in placental tissue in Menkes' disease. An immunohistochemical study.

Menkes' disease is a recessive X-linked disturbance of copper metabolism, resulting in accumulation of copper in several extra-hepatic tissues including the placenta. Metallothionein (MT) is a low-molecular weight protein with a high affinity for group II metal ions, such as copper. Its synthesis is induced by the presence of the ions. The aim of this study was to investigate the pattern of the MT immunoreactivity in placental tissue obtained from women at-risk of Menkes' disease in order to examine whether the MT occurrence and distribution may reflect the copper content. Placental tissue from six women with a family history of Menkes' disease, from 4 women without a family history, and from 2 hydatiform moles was studied. Positive MT immunostaining was found to be independent of the length of fixation, whether the tissue samples were fixed in 4% buffered formaldehyde or Bouin's fixative. The avidin-biotin-complex (ABC)-technique was used. The copper content was measured by neutron activation analysis (NAA). In all placental tissue sections positive MT immunostaining appeared only in the trophoblast and only in proliferating cells. In placental tissue sections obtained from foetuses and children affected by Menkes' disease an additional MT immunostaining appeared in the Hofbauer cells of the chorionic villi. This staining was associated with an increased content of copper as measured by NAA. We conclude that the immunohistochemical demonstration of MT reflects the copper content and may be useful in pre- and postnatal diagnosis of Menkes' disease.

T cell-independent B cell response to self-monosialogangliosides: primary response monoclonal antibodies.

Purified GM1 and GM2 gangliosides incorporated into liposomes were injected subcutaneously in BALB/c mice every 3-4 days after pretreatment of the animals with low-dose cyclophosphamide. Serum samples were collected at different intervals and tests by ELISA for the presence of anti-ganglioside antibodies. Four doses (50 micrograms each) were sufficient to raise a measurable primary type of response to GM1, while nine doses were required to obtain measurable IgM antibody titers to GM2. Three monoclonal antibodies (MAbs) wer generated by fusing splenocytes with mouse myeloma cells. The specificity of MAbs was determined by ELISA and HPTLC-immunostaining using a panel of purified glycolipids. The MAb designated E1 showed a high degree of specificity because it reacted only with N-acetyl GM2. Monoclonal antibody A3 reacted predominantly with GM2 and GM1, but also reacted moderately with the GM3 ganglioside. The epitope recognized by this MAb is suggested to be the trisaccharide sequence GalNAc beta 1-4(NeuAc alpha 2-3)Gal. The third MAb (F6) reacted strongly with GM1 but a weak reactivity was also observed with GD1b as well as with asialo-GM1, indicating that the terminal tetrasaccharide Gal beta 1-3GalNAc beta 1-4(NeuAc alpha 2-3)Gal- structure is probably involved in antigenic recognition. Formalin-fixed and paraffin-embedded tissue sections were stained with the E1 and A3 MAbs, using the avidin-biotin complex (ABC) technique. Strong immunoreactivity for E1 appeared in the tumor cells of five primary lung carcinomas and in five malignant melanomas. No immunoreactivity was demonstrated in the parenchyma of a lung without malignancy, or in a metastasis from a colon carcinoma.

Immunohistochemical detection of p53 in epidermal proliferations overlying dermatofibromas.

Basaloid proliferations of epidermis overlying dermatofibromas and morphologically resembling superficial basal cell carcinomas might possess a malignant potential. In order to elucidate whether these basaloid proliferations share phenotypic characteristics with malignancy, we examined immunohistochemically 19 cases of dermatofibroma with overlying epidermal basaloid proliferations, 10 dermatofibromas with overlying simple epidermal proliferations, and 10 invasive basal cell carcinomas for expression of p53. Simple and basaloid proliferations showed sparse positive immunostaining for p53, as seen in normal epidermis. No differences in staining pattern or number of positive keratinocytes could be demonstrated between these conditions. The dermatofibromas were negative. The invasive basal cell carcinomas showed abundant p53 positivity. The lack of p53 immunoreactivity in the epidermal basaloid proliferations overlying dermatofibromas indicates that these lesions have not acquired a phenotype as seen in malignant conditions.

The prognostic significance of immunohistochemically detectable metallothionein in primary breast carcinomas.

Metallothionein (MT) is a low-molecular-weight protein with a high affinity for group II metal ions, especially zinc and copper. MT serves as an intracellular reservoir of these ions, but may also be involved in the detoxification of certain toxic metal ions such as cadmium. In addition, high MT contents might protect tumour cells from alkylating agents and irradiation. The aim of this study was to evaluate the prognostic significance of immunohistochemically detected MT overexpression in patients with primary breast carcinoma: 478 patients with primary breast carcinoma diagnosed during the period 1980-1985 were included. Formalin-fixed and paraffin-embedded tissue was used. Immunoreactivity for MT was found to be independent of the length of formalin fixation if the sections were microwave processed before incubation with the primary antibody. Patients were divided into two groups: those with MT overexpression (more than 10% positive tumour cells) and those with low expression (less than 10% positive tumour cells). MT overexpression was correlated with postmenopausal status, large tumour size, presence of lymph node metastases, high number of mitoses, severe nuclear pleomorphism, high histological grade (poor differentiation), and absence of PgR. In univariate analysis of survival data, MT overexpression was a predictor of poor overall survival in the entire group of patients. In multivariate analysis, MT overexpression failed to be of prognostic significance, whereas classical histopathological parameters such as tumour size, histological grade, and PgR were of independent prognostic significance.

An immunohistochemical study of p53 with correlations to histopathological parameters, c-erbB-2, proliferating cell nuclear antigen, and prognosis.

In an immunohistochemical study of 490 primary breast cancer patients with a follow-up period of more than 10 years, we found that p53 was not a prognostic factor for disease-free or overall survival among the whole cohort or among lymph node-positive or -negative patients. In a multiple logistic regression model classical histopathological parameters, such as lymph node status, number of mitoses, histological grade, and absence of progesterone receptors, were independent, poor prognostic predictors. In univeriate analysis p53 immunoreactivity was positively correlated with the absence of tubule formation, high histological grade (poor differentiation), absence of estrogen receptors (ER), and a high proliferating cell nuclear antigen (PCNA) score (ie, parameters indicative of an aggressive phenotype). The lack of prognostic significance may be attributable partly to the method used, because immunohistochemistry underdetects rather than overdetects p53 protein. No correlation between p53 and c-erbB-2-oncoprotein was demonstrated.

Immunohistochemical study of estrogen receptors in primary breast carcinomas and their lymph node metastases including comparison of two monoclonal antibodies.

Estrogen receptors (ER) status was investigated in 101 primary breast carcinomas and their axillary lymph node metastases to determine if the malignant cells retained or changed this phenotypic feature during the metastatic process. Immunohistochemistry with the ER-ICA kit (Abbott Laboratory) on formalin-fixed paraffin-embedded tissue was used (paraffin ER-ICA). The ER status in primary and secondary tumours was concordant in 80 patients (79%) and discordant in 21 (21%). Eighteen of these twenty-one patients had ER-positive primary tumours and ER-negative lymph node metastases. This discordance, which may be due to loss of ERs in the metastatic cells or tumour heterogeneity, could explain the well-known failure of endocrine treatment in some of the patients with ER-positive primary tumours. A new monoclonal antibody ID5 (DAKO) against ERs was applied on formalin-fixed paraffin-embedded tissue from 83 of these 101 primary carcinomas. These analyses and paraffin ER-ICA analyses were compared to prior analyses of the same 83 tumours using the ER-ICA kit on fresh frozen tissue ("gold standard", frozen ER-ICA). Kappa coefficient, sensitivity and specificity were 0.74, 0.96 and 0.75 for ID5 antibody, and 0.59, 0.72 and 0.96 for ER-ICA antibody on paraffin sections.

Proliferating cell nuclear antigen (pcna) and C-erbb-2 oncoprotein in breast-carcinoma with correlations to histopathological parameters and prognosis.

Overexpression of PCNA (more than 25% positive tumour cells) and positivity of c-erbB-2 oncoprotein were immunohistochemically demonstrated in 490 formalin-fixed and paraffin-embedded breast carcinomas. Overexpression of PCNA and c-erbB-2 correlated with large tumour size, presence of lymph node metastases, high histological grade (poor differentiation), and absence of steriod hormone receptors features indicating an aggressive phenotype. In univariate analysis overexpression of PCNA correlated with poor overall survival (p<0.05), whereas c-erbB-2 was of no prognostic significance. In multivariate analysis both PCNA and c-erbB-2 failed to be of independent prognostic significance. In order to identify women with different prognosis an index termed immunoscore, based upon the results of the immunoreactivity of both PCNA and c-erbB-2 was constructed. The immunoscore was correlated with tumour size, lymph node status, histological grade, and steroid hormone receptor status. In univariate analysis of survival data the immunoscore was a prognostic parameter of poor overall survival. In multivariate analysis the classical histopathological parameters such as tumour size, histological grade and progesterone receptor status turned out to be of independent prognostic significance. The immunoscore was associated with poor prognosis but did not reach independent statistical significance (p=0.08). Further studies including a larger number of patients must be carried out in order to determine the prognostic significance of the immunoscore in multivariate analysis.

Immunohistochemical detection of metallothionein in primary breast carcinomas and their axillary lymph node metastases.

Metallothioneins (MTs) are low-molecular-weight proteins with specific binding for group II metal ions. MTs are involved in the detoxification of metals, but can also play a role in protection of the cell against certain anticancer agents and from damage of irradiation. High expression of MTs in primary breast carcinomas has been found to be associated with poorer prognosis. Expression of MT (MT) was examined immunohistochemically in 160 breast carcinomas and their concomitant lymph node metastases. The immunoreactivity appeared to be independent of the length of fixation when the section was microwaved before incubation with the primary antibody, a monoclonal antibody E-9. The findings were correlated with various histopathological factors, disease-free survival and over-all survival. Patients were divided into two groups, those with MT over-expression (above 10% of positive tumour cells), and those with low MT expression (below 10% positive). MT over-expression was found to be correlated with postmenopausal status and inversely with positive progesterone receptor status (PgR). MT over-expression showed statistically significant correlation with poor over-all survival. No differences in survival were seen between pre- and postmenopausal patients. PgR was in univariate analysis a poor prognostic parameter. In one fourth of the patients, the lymph node metastases showed increased MT expression compared with the primary tumour. These patients had a poorer, but not statistically significant different survival. MT expression was not correlated to chemo- or radiation therapy.(ABSTRACT TRUNCATED AT 250 WORDS)

c-erbB-2 oncoprotein is not an independent prognostic parameter in primary breast carcinoma. An immunohistochemical study.

The prognostic significance of c-erbB-2 oncoprotein expression was studied using the monoclonal antibody, anti-c-erbB-2 (CB-11, BioGenex) and the avidin-biotin-complex (ABC) technique. Four hundred and ninety patients with primary breast carcinoma diagnosed at Gentofte Hospital in the period 1980-1985 were included. Information about treatment, relapse-free period and overall survival was obtained from the Danish Breast Cancer Co-operative Group (DBCG). The mean follow-up period was more than 10 years. Fifteen per cent of the tumours showed positive immunoreactivity for c-erbB-2. Oncoprotein expression was correlated with presence of lymph node metastases, type of tumour, high number of mitoses, severe nuclear pleomorphism, high histological grade (poor differentiation), and absence of steroid hormone receptors. By univariate analysis, expression of c-erbB-2 oncoprotein, was correlated with poorer overall survival and shorter disease-free period in the entire cohort and in patients with lymph metastases, but not in the group of patients without lymph node metastases. By multivariate analysis, c-erbB-2 failed to be an independent prognostic marker for either disease-free period or overall survival, whereas classical histopathological parameters such as presence of lymph node metastases, high number of mitoses, high histological grade (poor differentiation) and absence of progesterone receptors turned out to be of independent prognostic significance.

Microwave processing for immunohistochemical demonstration of proliferating cell nuclear antigen (PCNA) in formalin-fixed and paraffin-embedded tissue.

The effect of microwave irradiation on the immunoreactivity of proliferating cell nuclear antigen (PCNA) in five primary breast carcinomas, a tonsil with hyperplasia, and a seminoma of the testis was evaluated in formalin-fixed and paraffin-embedded sections using the monoclonal antibody PC-10. The tissues were fixed in 4% buffered formaldehyde for 6, 24, 28, 72 h, and 1 week. Before incubation with the primary antibody, sections were microwaved for x2 5 min in distilled water, in citrate buffer, or they were processed routinely without microwave irradiation. In sections microwaved in distilled water the immunoreactivity was found to be independent of the length of fixation. PCNA immunoreactivity in sections microwaved twice in citrate buffer was not satisfactory. The PCNA immunoreactivity in sections which were routinely processed without microwave irradiation was markedly decreased after 48 h of fixation, and was nearly absent in sections fixed for up to 1 week. In sections processed without microwave irradiation the optimal dilution of the primary antibody was 1:10, whereas in sections microwaved twice in distilled water the optimal dilution was 1:600. The morphology of the tissue was well preserved after microwave processing. We therefore recommend microwave processing in distilled water for x2 5 min for demonstration of PCNA in formalin-fixed and paraffin-embedded sections using the monoclonal antibody PC-10, since this technique yields optimal staining reactions independent of the length of fixation.

Erythema elevatum diutinum: case report showing an unusual distribution.

A case of erythema elevatum diutinum showing an unusual distribution is reported. Hard nodules were located on the patient's fingertips and pulps while the extensor surfaces were not involved. The disease was initially misinterpreted based on both clinical and histologic examination. Dapsone therapy was followed by a remarkable recovery. The clinical and histologic features of erythema elevatum diutinum are summarized.

Proliferating cell nuclear antigen in breast carcinomas. An immunohistochemical study with correlation to histopathological features and prognostic factors.

Proliferating cell nuclear antigen (PCNA), was examined by immunohistochemistry in 509 breast carcinomas. The immunoreactivity was found to be independent of the length of fixation when the tissue sections were microwaved before incubation with the primary antibody. The PCNA immunoreactivity was assessed by two semi-quantitative methods, which were correlated but not exchangeable. The comedo type of intraductal carcinomas and invasive ductal carcinomas had a higher PCNA score than other types. Lymph node metastases had a significantly higher PCNA score than primary carcinomas. High PCNA immunoreactivity was correlated with the presence of lymph node metastases, absence of tubule formation, numerous mitoses, severe nuclear pleomorphism, high histological grade and absence of progesterone receptors (PgR). PCNA in lymph node positive tumours was correlated with tumour type, especially with ductal carcinomas, absence of tubule formation, high histological grade and absence of PgR, whereas PCNA in lymph node negative tumours was correlated with large tumour size, numerous mitoses, severe nuclear pleomorphism and high histological grade. Number of mitoses and nuclear pleomorphism were the two most important factors in predicting the PCNA score; the absence of PgR and nuclear pleomorphism were important in lymph node negative and positive tumours, respectively. In a univariate analysis high PCNA score was found to be correlated with shorter relapse-free period and poorer over-all survival.

The prognostic-significance of cathepsin-d in breast carcinomas - an immunohistochemical study.

Cathepsin D, a lysosomal protease, was examined by immunohistochemistry in 481 breast carcinomas. The immunoreactivity for cathepsin D was registered in the tumour cells, in the surrounding stroma, and in the macrophages and correlated with clinicopathological parameters. In univariate analysis of survival data, less than 25% positive tumour cells, any positivity in the stroma, and moderate or strong positivity in the macrophages were poor prognostic parameters. In multivariate analysis, all three parameters were of independent prognostic significance; therefore a cathepsin D index was constructed. In multivariate analysis the cathepsin D index was also found to be of independent poor prognostic significance both in patients with and without lymph node metastases. Progesterone receptor status was an independent prognostic parameter in patients with lymph node metastases. From the present study we concluded that the cathepsin D index was a independent poor prognostic parameter.

Sudden onset of adverse effects due to low-dosage bleomycin indicates an idiosyncratic reaction.

We report a case of possible idiosyncratic reaction in a fifty-two-year-old man with centroblastic malignant lymphoma. Immediately after administration of 20 mg bleomycin intravenously for the first time, the patient developed stomatitis. When readministered a few weeks later the patient developed mucocutaneous manifestations such as stomatitis, symmetrical linear maculopapular exanthema on the neck, trunk, and both arms, and indurated nodules on the palms.

Expression of cytokeratin and erbB-2 oncoprotein in Paget's disease of the nipple. An immunohistochemical study.

The occurrence of epithelial membrane antigen (EMA), low molecular weight cytokeratin (CK) and erbB-2 oncoprotein was investigated in twenty-eight cases of Paget's disease of the nipple (PD) to determine their diagnostic usefulness. The ABC technique with monoclonal antibodies was used on formalin-fixed, paraffin-embedded tissue. The Paget's cells showed positive immunoreactivity for all three antigens investigated in a high percentage of PD. Immunoreactivity for CK and erbB-2 oncoprotein was restricted to the Paget's cells, whereas EMA in some cases also stained the adjacent keratinocytes. Since CK and/or erbB-2 oncoprotein occurred in 93% of the cases, we conclude that demonstration of both antigens is useful in the diagnosis of PD. ErbB-2 oncoprotein was also found to be expressed in a high percentage of the underlying intraductal and invasive carcinomas.

[Extracardiac rhabdomyomas]. / Ekstrakardiale rabdomyomer.

Two cases of extracardial rhabdomyomata are described. One case was discovered incidentally at autopsy while the other was primarily examined by needle aspiration biopsy. As far as the authors are aware, this is the first case to be reported.

[Naproxen versus indomethacin as night-time medication for patients with rheumatoid arthritis]. / Naproxen versus indometacin som natlig behandling til patienter med reumatoid arthritis.

The investigation consisted of a double-blind cross-over study of the effect of 75 mg indometacin, 500 mg naproxen or a placebo in 63 patients with rheumatoid arthritis accompanied by night pain and morning stiffness. All the patients received day treatment with 250 mg naproxen b.i.d. Only a few patients benefitted from the treatment at night and no differences in the effect of indometacin and naproxen were observed. Naproxen was better tolerated than indometacin. Day treatment with naproxen, which has a relatively long half life time decrease the need for supplementary treatment at night.