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H Mullink

Publications and source records attributed to H Mullink.

At least 37 records · Page 2Linked to original sources
PubMed1988

Expression of intermediate filament proteins in adrenal cortex and related tumours.

The intermediate filament profile of adrenal cortex and its related tumours has been evaluated. Most adrenocortical cells contained cytokeratin 8 and 18 as demonstrated by monoclonal antibodies CAM 5.2, M20, M9 and RGE53. Cytokeratin immunoreactivity was not confined to a functional zone of the adrenal cortex. Only a small number of the adrenocortical cells showed vimentin immunoreactivity. From normal adrenal cortex through adenomas, to carcinomas, there is a progressive decrease or even loss of cytokeratin immunoreactivity and an increase in vimentin immunoreactivity. Aberrant cytokeratin expression was not found in adrenocortical adenomas and carcinomas with the antibodies used. Awareness of the possible absence of cytokeratin immunoreactivity in adrenocortical carcinomas is important whenever antibodies to cytokeratins and vimentin are used for diagnostic purposes in poorly differentiated neoplasms.

Adenoma↗
PubMed1988

The expression of intermediate filaments and mam-6 antigen in relation to the degree of morphologic differentiation of carcinoma of the head and neck: diagnostic implications.

In this study, the immunoreactivity of several cytokeratin antibodies; 115 D8, a monoclonal antibody against MAM-6, an epithelial membrane antigen; and two vimentin antibodies, is examined in relation to the degree of morphologic differentiation in carcinomas of the head and neck. The results indicate that a relationship exists between the degree of morphologic differentiation and the expression of cytokeratin, MAM-6, and vimentin, as detected by polyclonal antikeratin, 115 D8 and anti-vimentin. Expression of cytokeratin and MAM-6 is reversely related to vimentin. Polyclonal anti-keratin; CAM 5.2, a monoclonal antibody against cytokeratin 8, 18 and 19; and 115 D8, used in combination, were still able to identify the epithelial nature of undifferentiated/spindle cells. Since these immunohistochemical markers precede light microscopic detectable signs of epithelial differentiation, they can be used for the identification of the epithelial nature of undifferentiated/spindle tumors of the head and neck.

Antigens, Differentiation↗
PubMed1988

Sensitivity of in situ detection with biotinylated probes of human papilloma virus type 16 DNA in frozen tissue sections of squamous cell carcinomas of the cervix.

The sensitivity of human papilloma virus type 16 (HPV-16) DNA detection by DNA in situ hybridization using biotinylated probes (bio-DISH) was estimated by performing this technique on snap-frozen tissue sections of 10 cervical squamous cell carcinomas containing increasing amounts of HPV-16 as determined by Southern blot hybridization. A protocol using serial sections for bio-DISH and DNA extraction was used. The number of positively stained cells and the detection limit were strongly dependent on the treatment of the sections with proteinase K prior to hybridization. At low proteinase K concentration (0.1 micrograms/ml), the detection limit appeared to be 30-40 HPV-16 DNA copies per carcinoma cell, whereas morphology was preserved. A high proteinase K concentration (1-5 micrograms/ml) often resulted in an increase in the number of positively stained cells but also in a poor morphology. The detection limit was improved to at least 20 HPV-16 DNA copies per carcinoma cell.

Biotin↗
PubMed1987

A rapid and simple hapten conjugation method for monoclonal antibodies to be used in immunoenzyme single and double staining procedures.

A rapid and simple method was developed for the haptenization of monoclonal antibodies (MAbs), to be used in immunoenzyme single and double staining techniques. Using this method minute amounts of MAbs can be haptenized without purification of the antibody or removal of the excess hapten. The haptenized MAb can be ready for use with 2.5 h. The method consists of a direct incubation of the antibody with the haptenizing agent and subsequent addition of an amino acid solution to stop the reaction. Commonly available reagents were tested, of which dinitrofluorobenzene, trinitrobenzene sulfonic acid and an oxazolone derivative gave the best results. The procedure was evaluated by using MAbs directed against lymphoid cell surface membrane antigens in an indirect immunoenzyme staining on frozen sections using peroxidase or alkaline phosphatase-conjugated anti-hapten antibodies as second step antibody. It was found that those MAbs, which show good staining results in conventional indirect immunoenzyme procedures, can also be used successfully after haptenization in single as well as double staining procedures. Combination of haptenized and biotynilated MAbs gave good results when weakly reactive MAbs had to be included in immunoenzyme double staining.

Antibodies, Monoclonal↗
PubMed1987

Expression of cytokeratins and vimentin in epithelial cells of normal and pathologic thyroid tissue.

The presence of intermediate filament proteins of the cytokeratin and vimentin type was evaluated in normal and pathologically changed thyroid tissue specimens. Using the indirect immunoperoxidase technique with 4 different cytokeratin monoclonal antibodies: RCK114 (broad spectred), K2080 (broad spectred), RGE53 (directed against component 18, present in simple epithelium) and RKSE60 (directed against component 10, associated with keratinization). Co-expression of cytokeratin and vimentin was evaluated with a double immunoenzyme staining technique. The results indicate that normal and transformed cells express cytokeratins of the non-epidermal type. Cytokeratins of the epidermal type are sometimes present in carcinomas. They do not differentiate in tumour type (i.e. papillary, follicular, anaplastic or medullary carcinoma). The co-expression of cytokeratins and vimentin is not restricted to carcinomas: in a small percentage of cases it is also present in normal epithelial cells of the thyroid gland. Moreover, the distribution pattern of cytokeratins and vimentin within the cell is changed in malignant transformed epithelial cells of the gland and seems to be inversely related to the degree of differentiation of these cells. The implications of our findings for the possible use of cytokeratins and vimentin in diagnostic pathology are discussed.

Antibodies, Monoclonal↗
PubMed1987

Classification of routinely processed anaplastic large cell tumours with a small panel of antibodies. An immunohistochemical study with clinical follow-up.

A proportion of anaplastic large cell tumours is difficult to classify on sections of routinely processed, paraffin-embedded tissue. Differentiation into large cell lymphoma, carcinoma, melanoma or sarcoma is important in order to assess prognosis and proper treatment. Although the use of immunohistochemistry has been reported in the differentiation between some of these types of neoplasms, no antibody panel, which can directly differentiate all of them, has been described. In the present study we evaluated the value of a panel of 5 antibodies for the classification of 29 anaplastic large cell tumours, which could not be classified by experienced pathologists using conventional histological and histochemical techniques. The panel, which can be used on routinely fixed paraffin-embedded tissue, consisted of 5 different antibodies directed against keratin, vimentin, the human milk-fat globule membrane antigen MAM-6, a melanoma associated antigen and common leucocyte antigen. The use of this panel directly resulted in a definite diagnosis in 95% of the cases and provided valuable information for the diagnosis in the remaining cases. The diagnosis was confirmed by additional marker studies and electron microscopy. Moreover, clinical follow-up, including treatment data, was in accordance with the diagnosis based on the panel.

Aged↗
PubMed1986

Simultaneous immunoenzyme staining of vimentin and cytokeratins with monoclonal antibodies as an aid in the differential diagnosis of malignant mesothelioma from pulmonary adenocarcinoma.

The occurrence and coexpression of the cytoskeletal proteins vimentin and cytokeratins were studied in malignant mesotheliomas and pulmonary carcinomas. For this purpose a double immunoenzyme staining with monoclonal antibodies was developed which made it possible to visualize vimentin and cytokeratins simultaneously within the same cell. A clear distinction between stromal cells (vimentin only) and tumour cells was also obtained. A total of 12 mesotheliomas (six mixed type and six epithelioid type) and 13 carcinomas (eight adenocarcinomas and five large cell undifferentiated carcinomas) were studied. The results revealed a clear difference between mesotheliomas and adenocarcinomas: 11 of 12 mesotheliomas showed coexpression of vimentin and cytokeratins in at least 50% of the tumour cells, while in seven of the eight adenocarcinomas none or only a few cells could be seen with this coexpression. In the undifferentiated large cell carcinomas three of five expressed both components, but in less than 25% of the cells. It is concluded that a reliable double immunoenzyme staining of vimentin and cytokeratins can be used as an additional means to distinguish malignant mesothelioma from pulmonary adenocarcinoma.

Adenocarcinoma↗
PubMed1986

Distribution of T-lymphocyte subsets in different portions of sarcoid granulomas: immunohistologic analysis with monoclonal antibodies.

The numbers and the distribution of T-lymphocyte subpopulations in lymph node granulomas from 11 patients with sarcoidosis were studied in cryostat sections by an immunoperoxidase technique. Greater numbers of helper T lymphocytes (Leu-3+) were found at the periphery than in the central portion of the same granuloma. Most of the suppressor T lymphocytes (OKT8+) were present at the periphery of the granulomas. In addition, the Leu-3a/OKT8 ratio varied from 0.7 to 1.8 in the outer compartment, while in the central portion of the granuloma much higher values (3 to 20) were found. These different distribution patterns of T-lymphocyte subsets provide evidence for two different compartments with different immune reactions in sarcoid granulomas.

Adult↗
PubMed1986

Radionuclides in detecting active granuloma formation. Gallium-67 scintigraphy and histopathology with autoradiographic findings.

Granuloma formation studies were performed on lungs of guinea pigs sensitized with FCA over 2 to 17 months. Prolonged time of sensitization revealed more granulomatous pulmonary tissue. An intravenous booster of FCA in the animals that had been sensitized for 3 months yielded enhanced granuloma formation within 5 days. The histopathology of these lungs was comparable with that seen in lungs of animals after 17 months of sensitization without booster. Enhanced local proliferation of macrophages, measured by [3H]thymidine incorporation and autoradiography, was seen in the lungs of the animals that had received boosters. Moreover, 67Ga scintigraphy was strongly positive in these animals. Scintigraphy of cell suspensions of pulmonary tissue from these animals showed that 67Ga was predominantly taken up (quantitatively as well as qualitatively) by the alveolar macrophages. Cell suspensions of sarcoidosis patients, prepared in the same way, showed only a low level of 67Ga uptake, one comparable to that of the pulmonary cell suspensions of the sensitized animals that had not received boosters. It is suggested that a negative scintigraphy in patients with chronic pulmonary granulomatous disorders could be (partly) explained by the absence of activated macrophages.

Animals↗
PubMed1985

The involvement of dendritic cells in the handling of the immune stimulant C. parvum. A morphological investigation using immunoperoxidase techniques.

In earlier experiments we showed that locally administered Corynebacterium parvum (C. parvum) stimulated the T-cell system and had a beneficial effect on the recurrence rate of surgically resected cancers of the uterine cervix. In this paper we report the use of an immunoperoxidase technique to trace C. parvum antigen in the draining lymph nodes. In a guinea pig model the popliteal lymph node was studied after the injection of 70 micrograms of C. parvum in the hind footpad. At 6 h, intact bacteria were detected in sinus histiocytes. A transient granuloma formation was apparent between days 2 and 6, originating in the subcapsular and interfollicular areas. Three antigen-positive cell types were observed in these granulomas: a) cytophagocytic macrophages which were weakly positive, the antigen being distributed in clumps; b) dendritic cells with a strong, fine-granular positivity and c) some epithelioid cells with a small positive cytoplasmic rim. The majority of epithelioid cells was negative. Antigen-positive dendritic cells were also observed just beyond the granulomas in the T-dependent paracortical area. These cells are known as interdigitating cells (IDC) and present antigen to T-cells. Ten days following C. parvum injection the lymph node follicles became positive and the antigen could be detected in the long cellular protrusions of the dendritic reticulum cells (DRC). DRC probably play a part in immunological memory by trapping antigen in the form of immune complexes.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1985

Influence of fixation and decalcification on the immunohistochemical staining of cell-specific markers in paraffin-embedded human bone biopsies.

A number of fixation and decalcification procedures were evaluated to determine their suitability for immunohistochemistry on trephine samples of bone marrow after paraffin embedding. In particular, the immunoreactivity of antigens characteristic for various hematopoietic cell lines (immunoglobulin heavy and light chains for plasmacytoid cells; elastase for neutrophil myeloid cells; lysozyme, alpha-1-antitrypsin and alpha-1-antichymotrypsin for hystiocytic cells; leukocyte common antigen for lymphocytes; hemoglobin and glycophorin A for erythroid cells; Factor VIII-related antigen for thrombocytoid cells) as well as some antigens specific for epithelial tumors (CEA, 115D8, and keratin) were investigated. Fixation in a mercuric chloride-formaldehyde mixture followed by decalcification in acetic acid-formaldehyde-saline proved to be the best procedure for antigen preservation and retention of morphologic detail. Moreover, there is no need of trypsinization when using this procedure. The only exception was Factor VIII-related antigen in megakaryocytes, which was best demonstrated in trypsin-digested sections of formalin-fixed and acetic acid-decalcified biopsies.

Antigens↗
PubMed1985

Veiled cells in the gastrointestinal tract.

Lately it has become known that for the initiation of an immune response an antigen presenting cell is needed. In the skin the antigen presenting cell system has been studied and is extensively supposed to consist of Langerhans' cells in the epidermis and dermis, veiled cells in the skin lymph, and interdigitating cells in the skin lymph nodes. These cells are considered to belong to the mononuclear phagocyte system which is in principle composed of cells stemming from the promonocyte/monocyte. To detect whether an antigen presenting cell system is present in the gut, we studied cell suspensions of gut tissues. Monoclonal cells with long actively moving cytoplasmic veils were found in cell suspensions from small intestinal villi of guinea pig and pig, but not of rat. These veiled cells appeared to be strongly Ia positive, as are the cells from the skin antigen presenting system. They share several ultrastructural and enzyme cytochemical characteristics with these skin cells. Gut veiled cells localize within the lympho-epithelium of Peyer's patches of the rat, just underneath this lympho-epithelium and in the T-dependent interfollicular area, according to Ia stainings on frozen tissue sections and to ultrastructural observations. From stainings with anti-IgA, -IgE, -IgG, -IgM and anti-T lymphocyte sera it is concluded that these cells do not belong to the lymphocyte series. It is suggested that these gut veiled cells belong to the monocyte/macrophage family and form an antigen presenting cell system in mucosa-associated lymphoid tissue, similar to the antigen presenting cell system in the skin.

Animals↗
PubMed1985

Immunocytochemical localization of pepsinogen I and II in the human stomach.

An immunoperoxidase method using antisera specific for PG I and PG II was developed to assess PG-containing cells in the human stomach. Fundus-chief cells contained abundant PG I and less PG II. Mucus neck cells stained more frequently with anti-PG I, while antral gland cells stained only with anti-PG II. In adenocarcinoma of the corpus, isolated glands faintly staining with anti-PG I or anti-PG II were observed. In dense tumorous tissue no anti-PG I activity could be observed, while PG II sometimes remained present in residual antral glands, even in areas of heavy tumor infiltration. Focal PG II activity was also clearly demonstrated in some antral tumor cells. The immunoperoxidase technique described is suitable for further detailed studies of normal and pathological conditions of the gastric mucosa.

Adenocarcinoma↗
PubMed1984

Protection against paracetamol-induced hepatotoxicity by acetylsalicylic acid in rats.

Acetylsalicylic acid (ASA) given simultaneously with paracetamol decreased paracetamol-induced hepatotoxicity (measured by plasma transaminase activities as well as histology) without any effect on glutathione depletion, indicating that ASA prevents a process (or processes) subsequent to the metabolic activation of paracetamol. Delayed treatment with ASA also reduced paracetamol-induced liver toxicity, suggesting that reduction of the absorption rate of paracetamol does not contribute essentially to the protection by ASA. Combinations of paracetamol and ASA may have potential use in the development of safer analgesic combinations containing paracetamol (or ASA).

Acetaminophen↗
PubMed1983

A quantitative histomorphologic analysis of lymph node granulomas in Sarcoidosis in relation to radiological stage I and II.

In a detailed study of granulomas in lymph nodes of 19 patients with Sarcoidosis we assessed and correlated a number of features, with size, development and radiographic staging of the disease. Except with the epithelioid cells the number of constituent cells (lymphocytes, giant cells and polymorphs), size, confluency and degree of necrosis of the granulomas were found to be very variable within a single gland. This indicates that the granulomas are in different stages of activity at a given time. All the features studied showed little or no correlation with the radiographic staging, which suggests that the latter is not a good index of the extent of the disease.

Adult↗
PubMed1983

Large mononuclear Ia-positive veiled cells in Peyer's patches. I. Isolation and characterization in rat, guinea-pig and pig.

The antigen presenting cell system in the skin is extensively studied, and is supposed to consist of Langerhans cells in epidermis and dermis, veiled cells in skin lymph and interdigitating cells in skin lymph nodes. In order to detect whether a similar cell system is present in the gut, we studied Peyer's patch cell suspensions. Mononuclear cells with long actively moving cytoplasmic veils were found in cell suspensions from Peyer's patches of rat, guinea-pig, and pig, and in cell suspensions from small intestinal villi of guinea-pig and pig, but not of rat. These veiled cells are strongly Ia-positive. Because of their Ia positivity, their enzyme cytochemical staining pattern, and their ultrastructure, these cells resemble the antigen presenting cells of the skin, skin lymph and lymph node.

Animals↗
PubMed1982

Histologic distinction between malignant mesothelioma, benign pleural lesion and carcinoma metastasis. Evaluation of the application of morphometry combined with histochemistry and immunostaining.

Thirty men and 7 women with malignant mesothelioma seen at the Free University Hospital from 1st January 1960 until 1st July 1981 were reviewed. The histological, histochemical and morphometrical findings are reported. These findings are compared with 25 cases of pleural metastatic carcinoma and 25 cases of reactive pleural lesions. Fourty-nine percent of malignant mesotheliomas produced hyaluronic acid, however all cases of pleural metastatic carcinomas failed to produce this substance. All cases of malignant mesothelioma were D-PAS negative while 15 cases of pleural metastatic carcinoma showed reactivity to D-PAS. All cases of malignant mesothelioma and 9 cases of metastases were CEA negative. To distinguish malignant mesothelioma from metastases it is advisable to perform the D-PAS staining first. If it is negative mesothelioma can be confirmed by showing hyaluronic acid activity. A positive CEA staining rules out mesothelioma. In our study it was shown that with these methods 18 of 37 mesotheliomas could be identified with certainty, and 22 of the 25 carcinoma metastases. Morphometrically the malignant mesotheliomas could not be distinguished from the metastases, however the reactive pleural lesions had smaller nuclei than the malignant cells with mean values below 30 mu2. In the malignant cases these values had a range from 36 to 101 mu2. In distinguishing between reactive pleural lesions and malignant mesothelioma the production of hyaluronic acid points to the malignant character of the lesion. Thus histochemistry and immunostaining are important in the distinction of malignant mesothelioma from metastases, while the value of morphometry lies mainly in the seperation of reactive lesions from malignant mesothelioma.

Aged↗