Ber-EP4 immunoreactivity depends on the germ layer origin and maturity of the squamous epithelium.

AIM: To map the expression of Ber-EP4 in well-differentiated squamous epithelia, metaplastic squamous epithelia and dysplastic squamous epithelia of different origins. METHODS AND RESULTS: Squamous epithelium of different origin was stained using a standard immunohistochemistry method applied to paraffin sections. We found that normal squamous epithelium of the oral cavity, oesophagus, uterine cervix, vagina, anal canal, and branchial cysts are Ber-Ep4-negative, as are the mature squamous metaplasia of bronchial mucosa, urinary bladder mucosa and uterine cervical mucosa. In contrast, immature squamous metaplasia of bronchial mucosa, or uterine cervical mucosa, and squamous dysplasia of oral mucosa of endodermal origin, or uterine cervical mucosa in most cases expressed Ber-EP4. CONCLUSION: Squamous epithelia of ectodermal origin never express Ber-EP4, whether normal, hyperplastic, dysplastic or neoplastic. In contrast, squamous epithelium of endodermal origin sometimes contains the target glycoproteins of Ber-EP4 when immature, metaplastic, dysplastic or neoplastic. The results indicate that the differences in expression of Ber-EP4 in squamous epithelium depend primarily on germ layer origin, and on the maturity of the epithelium.

Erythrokeratolysis hiemalis (keratolytic winter erythema): a case report from Denmark.

Erythrokeratolysis hiemalis is described as a rare autosomal dominant genodermatosis as first reported from the Oudtshoorn district of Cape Provence in South Africa, among European immigrant families. Sporadic cases have since been identified in other countries, often with a familial link to the Oudtshoorn cluster. We describe the first case observed in Denmark, presumably the result of a spontaneous mutation.

The terminal complement complex is generated in chronic leg ulcers in the absence of protectin (CD59).

Loss of membrane complement regulators accompanied by complement activation is suggested to be involved in the pathophysiological processes leading to tissue damage in myocardial ischaemia. In the present study we have investigated whether the same phenomenon may occur in ischaemic and/or venous hypertension leg ulcers. The deposition of complement, plasma complement regulators and expression of membrane regulators were detected by immunohistochemical methods, including immunofluorescence with antibodies against C3d, the terminal complement complex (TCC), vitronectin, clusterin, decay-accelerating factor (CD55) and protectin (CD59). Eleven frozen biopsies from ischaemic leg ulcers, 10 biopsies from venous hypertension leg ulcers, and 10 biopsies from normal skin were studied. In 9 of 11 ischaemic and in 5 of 10 venous hypertension leg ulcers, marked staining for TCC was found around the capillaries, most often at the ulcer margin. No TCC staining was found in normal skin. Staining for TCC was always accompanied by staining for clusterin and vitronectin and C3d. In normal skin, CD59 was found on the elastic fibers in the dermis, on the muscle coat, the Schwann sheath and acinar cells. Semiquantitative measurement of CD59 showed marked increased staining intensity in the endothelium in venous hypertension ulcers and diminished intensity in ischaemic ulcers compared to normal skin. No such difference could be observed for CD55. When TCC was positive in the capillary walls, weak or no staining for CD59 was found. A significantly higher ratio of TCC/CD59 was found in the ischaemic compared to venous ulcers (p = 0.018). This was due to a marked difference between the ulcer margins (p = 0.013). Localized areas in the venous ulcers had the same pattern as that seen in the ischaemic ulcers. Our results suggest that loss of CD59 may enhance deposition of TCC and that complement-dependent inflammation may be an important factor in the tissue-damaging processes seen in chronic leg ulcers.

Ulcer bed infection. Report of a case of enlarging venous leg ulcer colonized by Pseudomonas aeruginosa.

We report a case of ulcer bed infection in an enlarging venous leg ulcer without clinical signs of cellulitis in the surrounding tissues. Signs of infection in the leg ulcer were: 1) cocci-like structures and bacteria-like rods around vessel walls in the viable ulcer bed, 2) vasculitis-like inflammation of deeply situated vessels of the viable tissue, 3) Pseudomonas aeruginosa-specific antibodies in the serum (other than against exotoxin A), 4) extensive epidermolysis of normal human skin by the wound exudate in vitro, and 5) P. aeruginosa exotoxin A in the wound exudate (23 ng/ml). In an in vitro cell assay, the wound exudate was cytotoxic and rabbit antibodies to exotoxin A, but not a serine proteinase inhibitor, inhibited this cytotoxicity. P. aeruginosa exotoxin A might contribute to the pathogenesis of the ulcer enlargement. The ulcer improved after the third skin graft, probably mainly due to effective treatment with a long-stretch compression bandage.

Patterns in expression of insulin-like growth factor-II and of proliferative activity in the normal human first and third trimester placenta demonstrated by non-isotopic in situ hybridization and immunohistochemical staining for MIB-1.

The expression of insulin-like growth factor-II (IGF-II) in normal human first and third trimester placental tissue was investigated by non-isotopic in situ hybridization (ISH). This is the first ISH study on IGF-II expression in placenta using an alkaline phosphatase-labelled probe. The expression was correlated with the proliferative activity of the cells using the proliferative marker MIB-1. In first trimester tissue, IGF-II was expressed in the cytotrophoblast, the extravillous trophoblast, the fetal endothelial cells and the mesenchymal fetal cells in the villi. In third trimester tissue, IGF-II expression was found in the amnion, the extravillous trophoblast and the mesenchymal fetal cells especially in the endothelial cells and the outer contractile sheet in the stem villi. In areas with perivillous fibrin deposits, strong expression of IGF-II was found in the cytotrophoblasts invading the fibrin. In first trimester tissue, the proliferative activity of the villous cytotrophoblast correlated well with the degree of IGF-II expression whereas in third trimester tissue, there was a discrepancy between MIB-1 positivity and the IGF-II expression. Expression of IGF-II does not seem to be correlated exclusively to the mitogenic activity of cells.

Epidemiology of classic Kaposi's sarcoma in Denmark between 1970 and 1992.

BACKGROUND: Studies have suggested that the incidence of classic Kaposi's sarcoma (KS) varies considerably within Europe. The epidemiology of classic KS in Denmark was described for the period between 1970 and 1992 and association with marital status and country of origin were evaluated. METHODS: A descriptive epidemiology study was carried out using data from the Danish Cancer Registry and from all Danish pathology departments. The effects of marital status and of immigration were evaluated in a case-control study with sex- and age-matched controls for 50 cases of classic KS. RESULTS: A total of 59 patients with classic KS, 34 men, 25 women, were identified between 1970 and 1992, yielding world standardized incidence rates of 0.40 and 0.22 per 1,000,000 population among men and women, respectively. Among men, a larger proportion of the patients with classic KS were of non-Danish origin [odds ratio (OR) = 3.8, 95% confidence interval (CI) = 1.5-9.6]. No excess of foreign born persons was observed among women with KS. Never-married men were at a markedly increased risk for classic KS (OR = 2.8, 95% CI = 1.2-6.7). This risk was, however, restricted to men younger than 60 years (OR = 18.8, 95% CI 3.4-104.8), whereas no increased risk was observed in men age 60 years or older nor was there an increased risk in women. CONCLUSIONS: Classic KS is a rare disease in Denmark. However, men, immigrants and never-married men age 60 years or younger experienced an excess risk. Even though the latter finding might reflect an association with male homosexuality, the observation should be interpreted with caution due to the limited number of observations.

Histological changes in the nasal mucosa after hot-water irrigation. An animal experimental study.

Many years ago the treatment of posterior epistaxis was irrigation with hot water through the bleeding nose cavity, and the treatment was successful in many cases. The aim of this study is to explain how "hot-water irrigation" can cause haemostasis. Twenty-four rabbits were divided into 12 groups and their noses were irrigated for 5 min with hot water at temperatures ranging from 40-60 degrees C. After irrigation, the nose was fixed, sliced, stained, and evaluated blindly by a pathologist. The morphological changes-narrowing of intranasal lumen, vasodilation and stasis, extravasation of erythrocytes, and epithelial necrosis-were recorded. No changes were recorded after irrigation with water of 40-44 degrees C. Only light changes were present in the 46 degrees C group. Vasodilation occurred at a temperature of 48 degrees C or higher. From 48 degrees C, oedema of the mucosa and subsequent narrowing of the intranasal lumen was seen. Severe changes including epithelial necrosis, were found only in the groups treated with 52 degrees C or higher. The results of the study indicate that the haemostatic effect of hot water treatment for epistaxis may be caused by: (1) oedema and narrowing of the intranasal lumen, (2) vasodilation of the mucosal vessels, and (3) cleaning of the nose from blood coagulates.

Ignored trichomonal infestation diagnosed by Papanicolaou smear.

OBJECTIVE: To compare the occurrence of Trichomonas vaginalis as demonstrated by culture and by Papanicolaou (PAP) smears in a sexually transmitted disease (STD) clinic. SETTING: The largest out-patient venereological clinic in Denmark. SUBJECT AND METHODS: As the prevalence of trichomonal infestation has decreased considerable in recent years direct microscopy of vaginal wet mounts is no longer performed routinely. Instead the screening diagnostic procedure is based on culture. We have retrospectively collected data on culture-negative women with Trichomonas vaginalis organisms present in cervical smears, taken on a routine basis to exclude atypical cells, and compared with the clinical findings. RESULTS: Since 1992 a total of 17 women were found to harbour Trichomonas vaginalis in cervical smear. A vaginal discharge was described in 10 women, six of whom had concomittant unspecific vaginitis. However, four women had unexplained vaginal discharge that could have been related to infestation with Trichomonas vaginalis. In addition one asymptomatic woman had a male partner with persistent urethritis. CONCLUSION: The prevalence of trichomoniasis is underestimated in women attending the clinic if the diagnosis is based on culture alone. PAP smears may be helpful in demonstrating characteristic trichomonal organisms. In general we do not recommend the PAP smear be used to diagnose STDs. However the finding of trichomanal organisms in smears should prompt a repeated culture and direct microscopy of vaginal wet mount.

The delayed-type hypersensitivity reaction is dependent on IL-8. Inhibition of a tuberculin skin reaction by an anti-IL-8 monoclonal antibody.

Cell-mediated immune reactions are essential to our immune response toward foreign organisms such as microorganisms, or in the response toward foreign tissue Ags, as seen in the rejection of allogeneic transplanted organs. Similar reactions form the basis for the development and the progression of delayed-type hypersensitivity (DTH) reactions. We found that the alpha-chemokine IL-8 plays an important pathophysiologic role for the development of a DTH reaction because infusion of a neutralizing anti-IL-8 mAb (WS-4) was able to suppress the development of a tuberculin skin reaction in rabbits, as judged by histologic, biochemical, and clinical examinations. Thus, the number of neutrophil granulocytes and lymphocytes at the site of tuberculin injection was decreased considerably, and the clinical signs of inflammation were suppressed almost completely at 24 h after intracutaneous injection of tuberculin, as judged by the size of the infiltrates. In contrast, we did not see any effect on the visible erythema of the skin. We found that the tissue content of myeloperoxidase (MPO), reflecting the number of infiltrating neutrophils, was lowered significantly. Furthermore, immunohistochemical analysis confirmed that IL-8 immunoreactivity is actually enhanced in the skin of positive tuberculin reactions. The results indicate that IL-8 plays an important role for the early accumulation of leukocytes in the skin and for the clinical signs of a DTH reaction.

Beta 2-microglobulin expression of AIDS-related and classical Kaposi's sarcoma.

The expression of beta 2-microglobulin, the invariable light chain of HLA class I molecules, of Kaposi's sarcoma from 11 AIDS patients and from 11 patients without known immunodeficiency was studied by immunohistochemistry using a polyclonal antibody to beta 2-microglobulin. The staining intensity of spindle cells in these lesions was scored in a semiquantitative system. We found that the spindle cells of Kaposi's sarcomas from AIDS patients showed significantly increased staining intensity for beta 2-microglobulin compared to those of Kaposi's sarcomas from non-AIDS patients. The results may indicate that Kaposi's sarcomas developing in immunocompromised individuals, such as AIDS patients, are not subject to immune selection by T cells eliminating HLA class I high-expressing tumor cells, while this may be the case in non-AIDS patients. Alternatively, the results may be caused by differences in the activity of cytokines, which upregulate the expression of HLA class I molecules on the cell surface.

[Dermatoscopy. An investigative method for the diagnosis of pigmented skin tumors]. / Dermatoskopi. En undersøgelsesmetode til diagnosticering af pigmenterede hudtumorer.

Dermatoscopy is a non-invasive investigative technique which makes it possible to evaluate the pigmented structures of the epidermis, the dermo-epidermal junction and the papillary dermal layer. The melanin pigmentation of the epidermal basal cell layers, the so-called pigment network is the primary target of dermatoscopy. A number of diagnostic criteria and variables for the dermatoscopy of pigmented skin lesions have been developed. Cutaneous malignant melanoma is still increasing in incidence and mortality. An early diagnosis is decisive for the prognosis. Therefore, it is urgent to develop practical investigative methods which can increase the diagnostic sensitivity. Dermatoscopy is such a method, but it demands training and experience. Lack of suspicious findings by dermatoscopy does not exclude malignancy. A safe diagnosis can only be obtained by a histological examination.

[Malignant lentigo melanoma]. / Lentigo maligna melanom.

Malignant melanoma (MM) arises either from a pre-existing, pigmented nevus, from clinically normal skin, or in rare cases from the non-nevoid lesion named lentigo maligna (LM). We present two cases of LM, one in a 75-year old woman and one in an 89-year old woman, which progressed to MM over ten and two years respectively.

The occurrence of pericapillary fibrin in venous hypertension and ischaemic leg ulcers: a histopathological study.

The presence of pericapillary fibrin and complement C3c in the ulcers of 19 patients with venous hypertension and 14 patients with ischaemic leg ulcers was investigated using histochemical and immunohistochemical techniques. There was deposition of fibrin around the capillaries in the central part of the ischaemic ulcers, and the venous hypertension ulcers, and in the non-ulcerated skin around one of the venous hypertension ulcers and two of the ischaemic leg ulcers. The deposition of fibrin is a secondary phenomenon that occurs in the area of ulcerated skin and does not play a major causal role in the formation of chronic leg ulcers.

Evaluation of the reproducibility of the revised 1988 International Federation of Gynecology and Obstetrics grading system of endometrial cancers with special emphasis on nuclear grading.

An intraobserver and interobserver analysis of the reproducibility of the revised 1988 International Federation of Gynecology and Obstetrics (FIGO) grading system of endometrial cancers was performed in 47 endometrial cancers (94 biopsy specimens). In the revised FIGO grading system the growth pattern still serves as the basic parameter, but nuclear atypia inappropriate for the architectural grade, raises the grade by 1. Ninety-four slides were graded twice by two pathologists. Analysis by the kappa statistics (corrected for agreement by chance) showed an acceptable result for interobserver reproducibility: kappa value 0.65 (95% confidence interval, 0.480 to 0.818). Further study showed this to be a reflection of the reproducibility of the architectural grade: kappa 0.70 (95% confidence interval, 0.565 to 0.850), whereas the reproducibility of the nuclear grade was poor: kappa value 0.55 (95% confidence interval, 0.383 to 0.713). Review of the literature showed great variability in the definition of nuclear grading, indicating that the revised 1988 FIGO grading system is not applicable as standard of reference for studies of endometrial cancers, before agreeing to the definition of inappropriate nuclear atypia.

Diagnosis of electrical skin injuries. A review and a description of a case.

This paper presents a brief review of the results of research in the area of electrical skin injuries. It also includes a case report of a 5-year-old girl noted on her admission to the hospital to have injuries to the skin of her chest and left arm. Histological examinations demonstrated that the skin lesions were segmental and showed necrosis and inflammation. Deposits of calcium salts distinctly located to collagen fibers were observed below the regenerated epidermis at the periphery of two skin lesions of the chest wall, in the lower part of dermis at the periphery of a skin lesion of the left arm, and within connective tissue adjacent to elastic arteries and peripheral nerves from the thoracic cavity. The pattern of calcification localized to collagen fibers and situated both superficially and deeply in the skin in a zone of viable tissue close to necrotic tissue is characteristic of electrically induced lesions. Although there have been reports of deposition of calcium salts on collagen fibers after application of calcium salts in high concentration at the skin surface, the collagen calcification in the pattern observed in this patient is probably diagnostic of electrical injury.

Reproducibility of histopathological evaluation in epithelial ovarian carcinoma. Clinical implications.

An interobserver analysis of the reproducibility of malignancy grade and of histological type classified according to World Health Organization criteria was performed in 261 ovarian tumours, stages IIB to IV. All available slides per patient including metastatic tumours (16%) were reviewed independently by two pathologists. Overall observed agreement regarding type was 68%. The chance-corrected agreement expressed by the kappa coefficient was 0.55 (95% confidence interval 0.49-0.61). Overall observed agreement regarding grade was 41% compared with a kappa coefficient of 0.19 (95% confidence interval 0.12-0.26). Kappa coefficients, ranging from 0.18 to -1.00 for associations between type categories and from 0.20 to -0.32 for grade categories, indicated that any suggested pattern of association could be caused by chance occurrences. A more objective approach to histopathology is needed in ovarian carcinoma before its prognostic impact can be assessed.