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  • 1990-1994  (8,592)
  • 1930-1934  (1,172)
  • 1920-1924  (688)
  • Cell & Developmental Biology  (9,837)
  • Immunohistochemistry  (654)
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  • 1
    ISSN: 1432-2307
    Keywords: Malignant fibrous histiocytoma ; Soft tissue sarcomas ; Immunohistochemistry ; Monoclonal antibody
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract An immunohistochemical study on frozen sections was carried out on 51 malignant tumours of soft tissue and bone using the FU-3 monoclonal antibody. This antibody is claimed to be specific for malignant fibrous histiocytoma (MFH) and liposarcoma and for normal and tumour cells located in perivascular fields. The results show a lack of specificity in MFH staining: several malignant tumours such as synovial sarcoma, fibrosarcoma, rhabdomyosarcoma, osteogenic sarcoma, and including an anaplastic malignant melanoma, presented positive staining somewhat similar to that found in MFH. The value of this antibody in the differential diagnosis of MFH is doubtful. It might be useful to recognize a common pathway of terminal differentiation expressed by several pleomorphic sarcomatous neoplasms.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Virchows Archiv 424 (1994), S. 267-271 
    ISSN: 1432-2307
    Keywords: Keratin ; Immunohistochemistry ; MNNG ; Oesophageal carcinoma ; Shrew
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The distribution of keratins in N-methyl-N'-nitro-N-nitrosoguanidine-induced oesophageal carcinomas in shrews was tested immunohistochemically, using a panel of seven different monoclonal antibodies. The studies were done on methacarn-fixed paraffin-embedded tissue, using the labelled streptavidin biotin method, and the relationship between morphological characteristics and keratin reaction patterns in carcinomas was analysed and compared with that in adjacent “normal” oesophageal epithelium. In the normal oesophageal epithelia, KL1, AE1, AE3, CK8.12 and CK4.62 stained suprabasal cells, 312C8-1 reacted to basal cells, and KS-1A3 labelled all epithelial cells. In squamous cell carcinomas, almost all the cancer cells were labelled strongly by 312C8-1 and weakly by KS-1A3, while a few cells in the centres of the keratinized foci were stained by KL1, AE1, AE3, CK8.12, and CK4.62. Like human oesophageal carcinomas, shrew oesophageal carcinomas maintain expression of human keratin 14, as determined by 312C8-1. The expression of human keratin 13, as determined by KS-1A3, was down-regulated.
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  • 3
    ISSN: 1432-2307
    Keywords: Cytomegalic inclusion disease ; Viral replication ; Viral regulatory proteins ; Immunohistochemistry ; In situ hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Postmortem tissues from infants with congenital and postnatally acquired human cytomegalovirus (HCMV) infection were examined by routine histology, immunohistochemistry (IHC) and in situ hybridization (ISH) to determine the dynamics of viral replication in vivo. Histologically, infants in both groups showed characteristic inclusion-bearing cells most commonly in lung, kidney, liver and pancreas. IHC for late proteins using a rabbit polyclonal antibody and ISH for viral genomes detected most of the infected cells as nuclear and/or cytoplasmic signals. However, immunostaining with a monoclonal antibody against viral immediate early (IE) proteins was variable depending on the stage of viral replication within an individual infected cell. In tissues of infants with postnatal HCMV infection, many cells harboured IE antigens, while in tissues from congenital cases most of the affected cells lacked IE antigens and only a few showed cytoplasmic staining. The difference was not caused by the antigenic diversity among viral strains as confirmed by in vitro study. Our findings suggested that congenital infections exhibited uniformly late stage proteins with inactive viral replication at death, while acquired ones remained active. The different viral activity may reflect the immune status of congenital and acquired HCMV infections.
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  • 4
    ISSN: 1432-2307
    Keywords: L-pyruvate kinase ; M2-pyruvate kinase ; Kidney neoplasms ; Carbohydrate metabolism ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Using immunohistochemical and enzyme biochemical methods we investigated the expression of L- and M2-pyruvate kinase (PK) in normal renal tissue, renal cell carcinomas (RCCs; of clear cell, chromophilic cell and mixed cell type) and RCC metastases. L-PK was expressed in the proximal tubules of normal renal tissue and, to a variable extent, in 23/25 primary RCCs, in 1 RCC recurrence and in 10 RCC metastases. Staining intensity and percentage of stained tissue did not correlate with tumour grade. One renal oncocytoma and all extrarenal malignancies examined lacked L-PK immunoreactivity. M2-PK was mainly expressed in the distal tubules of the normal kidney and was found in all renal tumours as well as extrarenal malignancies. Quantitative biochemical investigations yielded a two- to seventeen-fold increase in PK activity in RCCs compared to the normal renal cortex taken from the same patient, whereas fructose-1,6-bisphosphatase and cytosolic glycerol-3-phosphate dehydrogenase activity was dramatically lower in RCCs. Otherwise, the activity of all other enzymes investigated (glucose-6-phosphate dehydrogenase, enolase and lactate dehydrogenase) was not significantly changed in the RCCs. The immunocytochemical results suggest that L-PK is a useful marker for RCC and its metastases, if acetone-fixed tissue is available. The quantitative changes of the concentration of PK and other enzymes in RCCs when compared with normal renal tissue probably reflect metabolic alterations related to tumour growth.
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  • 5
    ISSN: 1432-2307
    Keywords: Pulmonary artery ; Neoplasm ; Sarcoma ; Immunohistochemistry ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Herein we report the clinicopathological features of four cases of pulmonary artery sarcoma that appeared at our institution during a period of 30 years. The patients, 2 males and 2 females, were 50–62 years old. Tumour was found in the pulmonary trunk and right pulmonary artery in all cases, in the pulmonary valve and left pulmonary artery in three of the four cases, and in the right ventricular outflow tract in one case. There was direct extension or metastases to the lungs in two cases, the heart in one case, mediastinum or lymph nodes in two cases and the pleura in one case. Ultrastructural examination in one case revealed cells with features of smooth muscle cells and myofibroblasts. Immunohistochemical examination of three cases gave the following results: vimentin and smooth muscle specific actin was positive in all three cases, desmin in one case and cytokeratin in one case. No positivity was found for Factor VIII. This and other studies indicate that histologically most pulmonary artery sarcomas are leiomyosarcomas or “undifferentiated spindle cell sarcomas”. Immunohistochemical and ultrastructural examinations favour an origin from myofibroblasts, probably derived from multipotent (undifferentiated) cells in the wall of the vessel. Most lesions show extensive intrathoracic growth although they rarely metastasize outside the thoracic cavity. They have a poor prognosis although some cases are currently being diagnosed during life.
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  • 6
    ISSN: 1432-2307
    Keywords: Extracellular matrix ; Immunohistochemistry ; Squamous cell carcinoma ; Invasiveness ; Metastasis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The expression of extracellular matrices (ECMs) laminin (LN), type IV collagen (IV C), heparansulphate proteoglycan (HS-PG), fibronectin (FN), tenascin (TN), decorin and vitronectin (VN) was examined immunohistochemically in 112 primary tumours and 29 metastatic cervical lymph nodes in oral squamous cell carcinoma (OSCC). In highly invasive primary tumours, the expression of LN, IV C and HS-PG in the basement membrane along the tumour-stroma borderline and the expression of decorin and VN in the tumour stroma at the invasive site were all significantly decreased. The expression of FN and TN in the tumour stroma at the same site was markedly increased. In peritumour stroma in metastatic lymph nodes, LN, IV C, HS-PG, decorin and VN were weakly expressed, while FN and TN were strongly expressed. Thus, the staining pattern of the ECMs in the metastatic lymph nodes was similar to that in highly invasive primary tumours. Furthermore, in primary tumours of metastatic cases, the expression of LN, IV C, HS-PG, decorin and VN obviously decreased, while the expression of FN and TN increased when compared with those of the non-metastatic cases. The investigation of ECMs in OSCC was valuable in predicting tumour behaviour.
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  • 7
    ISSN: 1432-2307
    Keywords: Cathepsin D ; Cathepsin E ; Rosai-Dorfman disease ; Langerhans' cell histiocytosis ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Nosological classification of sinus histiocytosis with massive lymphadenopathy (SHML; Rosai-Dorfman disease) is difficult, and the normal cellular counterpart of Rosai-Dorfman (RD) cells is uncharacterised. The peculiar S-100+ phenotype of RD cells suggests a relationship with the dendritic cell family. Recent investigations have revealed cathepsin E to be selectively concentrated in antigen-presenting cells, whereas cathepsin D was found to be expressed in cells of macrophage lineage. Cathepsin D and E distribution was investigated by immunohistochemistry in a series of SHML biopsies and in two types of dendritic cell proliferative lesions: dermatopathic lymphadenitis (DL) and Langerhans' cell histiocytosis (LCH). In SHML biopsies, RD cells and monocyte-related elements of the sinuses and pulp coexpressed cathepsin D and E. LCH cells also stained for both these aspartic proteinases. Conversely, in DL cathepsin E and D were localised to separate cells that resembled Langerhans' cells (LC) or macrophages, respectively, in morphology and distribution. Our data outline the peculiar immunophenotype of RD and LCH cells and suggest that caution should be exercised in the identification of their normal cellular counterpart. The common expression of cathepsin D and E and of S-100 protein suggests some phenotypic overlap between SHML and LCH cells, despite their striking morphological divergence.
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  • 8
    ISSN: 1432-2307
    Keywords: Bcl-2 protein ; Mesothelioma ; Pleura ; Immunohistochemistry ; Prognosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Immunohistochemical study of bcl-2 protein immunoreactivity in human non-neoplastic mesothelium (44 cases) and in malignant mesothelioma (62 cases) using a murine monoclonal antibody (clone 124) showed cytoplasmic immunoreactivity for bcl-2 protein in five cases of malignant mesothelioma. Non-neoplastic mesothelium was not immunoreactive. Immunoreactivity for bcl-2 protein does not add useful prognostic information in malignant mesothelioma since survival times of bcl-2 positive and bcl-2 negative cases did not differ. Nevertheless, the detection of bcl-2 protein in malignant mesothelioma might be useful for the differentiation from reactive mesothelium.
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  • 9
    ISSN: 1432-2307
    Keywords: Bcl-2 protein ; Mesothelioma ; Pleura ; Immunohistochemistry ; Prognosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Immunohistochemical study of bcl-2 protein immunoreactivity in human non-neoplastic mesothelium (44 cases) and in malignant mesothelioma (62 cases) using a murine monoclonal antibody (clone 124) showed cytoplasmic immunoreactivity for bcl-2 protein in five cases of malignant mesothelioma. Non-neoplastic mesothelium was not immunoreactive. Immunoreactivity for bcl-2 protein does not add useful prognostic information in malignant mesothelioma since survival times of bcl-2 positive and bcl-2 negative cases did not differ. Nevertheless, the detection of bcl-2 protein in malignant mesothelioma might be useful for the differentiation from reactive mesothelium.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1432-2307
    Keywords: Cathepsin D ; Cathepsin E ; Rosai-Dorfman disease ; Langerhans' cell histiocytosis ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Nosological classification of sinus histiocytosis with massive lymphadenopathy (SHML; Rosai-Dorfman disease) is difficult, and the normal cellular counterpart of Rosai-Dorfman (RD) cells is uncharacterised. The peculiar S-100+ phenotype of RD cells suggests a relationship with the dendritic cell family. Recent investigations have revealed cathepsin E to be selectively concentrated in antigen-presenting cells, whereas cathepsin D was found to be expressed in cells of macrophage lineage. Cathepsin D and E distribution was investigated by immunohistochemistry in a series of SHML biopsies and in two types of dendritic cell proliferative lesions: dermatopathic lymphadenitis (DL) and Langerhans' cell histiocytosis (LCH). In SHML biopsies, RD cells and monocyte-related elements of the sinuses and pulp coexpressed cathepsin D and E. LCH cells also stained for both these aspartic proteinases. Conversely, in DL cathepsin E and D were localised to separate cells that resembled Langerhans' cells (LC) or macrophages, respectively, in morphology and distribution. Our data outline the peculiar immunophenotype of RD and LCH cells and suggest that caution should be exercised in the identification of their normal cellular counterpart. The common expression of cathepsin D and E and of S-100 protein suggests some phenotypic overlap between SHML and LCH cells, despite their striking morphological divergence.
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  • 11
    ISSN: 1432-2307
    Keywords: Endothelin-1 ; Adrenal gland ; Adrenal tumour ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Endothelin (ET)-1 is a 21-amino acid peptide with potent vasopressor and vasocontrictive properties. Biochemical studies suggest that this peptide occurs in adrenal glands, where it influences steroid hormone production. However, we have found no report of the topographical distribution of this peptide. The localization of ET-1 immunoreactivity in non-neoplastic (37 cases) and neoplastic adrenal glands (48 cases) was investigated with a sensitive immunohistochemical technique applied to routinely processed tissue specimens. ET-1 immunoreactivity was regularly seen in the cortex, especially in the zona fasciculata and to a varying extent also in the other two zones, but not in the medulla. The immunoreactive material appeared in the cytoplasm mostly in the form of vacuolar structures but also as grains. Focally, the cell membrane also showed immunoreactive staining. In the zona reticularis the immunoreactivity appeared mainly as cytoplasmic grains. Most cortical adenomas displayed numerous immunoreactive cells. The immunoreactivity in the tumour tissue appeared in the same forms as in normal cortex, but the reactive products were generally fewer in number. No obvious differences in immunostaining were seen between the aldosterone- and cortisol-producing adenomas or the non-functioning ones. Three of the ten carcinomas contained immunoreactive cells, but they were few, appearing focally and the ET-1 immunoreactive structures were seen as ‘dust-like’ material. The difference in immunoreactivity between the benign and the malignant cortical neoplasms may be of diagnostic value. Functionally our results support a relationship between ET-1 and steroid regulation in non-neoplastic cortical tissue.
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  • 12
    Electronic Resource
    Electronic Resource
    Springer
    Virchows Archiv 425 (1994), S. 315-320 
    ISSN: 1432-2307
    Keywords: Giant cell reparative granuloma ; Solid aneurysmal bone cyst ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Giant cell reparative granuloma (GCRG) is a reactive bone lesion that most often involves the jaws. However, occasional cases of GCRG in the distal extremities have been reported, to which we add five cases. All the patients were young to middle-aged adults and had sharply bordered, lytic lesions. Histologically, all the lesions had areas of osteoclast-like giant cells and osteoblast mantled osteoid. Two of the cases had foci of osteoclast-like giant cells lining vascular spaces. In extragnathic locations, GCRG may simulate other osteolytic giant cells lesions such as giant cell tumour of bone and aneurysmal bone cyst (AnBC). Immunohistochemically, all cases showed positive staining of the stromal spindle cells for vimentin and actin, and of the osteoclast-like giant cells for CD68, vimentin and leucocyte common antigen. GCRG is a benign lesion and conservative therapy is curative. As GCRG may have histological features which resemble AnBC it may be considered to be the solid variant of AnBC.
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  • 13
    ISSN: 1432-2307
    Keywords: Breast neoplasms ; Apocrine glands ; Immunohistochemistry ; Hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The frequency and the significance of apocrine differentiation in carcinomas of the breast are uncertain, because of the lack of reliable and reproducible criteria for morphological diagnosis. The 15 kDa glycoprotein of cystic breast disease (GCDFP-15) is regarded as a specific functional marker of apocrine cells. Expression of the prolactin-inducible protein (PIP)/GCDFP-15 gene was investigated by Northern blot analysis and in situ hybridization in breast cancer cell lines and in an unselected series (33 cases) of primary carcinomas of the breast. On the same cases, histological assessment of apocrine differentiation and immunocytochemical detection of GCDFP-15 were also performed and correlated with follow-up data. The presence of PIP/GCDFP-15 mRNA was a feature of a relatively high number of cases, but was incompletely correlated with histological and immunocytochemical evidences of apocrine differentiation. Expression of the PIP/GCDFP-15 gene was significantly associated with relapse-free survival, and may represent a novel variable of functional and prognostic relevance.
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  • 14
    ISSN: 1432-2307
    Keywords: Primary retroperitoneal mucinous cystoadenocarcinoma ; Immunohistochemistry ; K-ras mutation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Special immunohistochemical stains for the identification of gastroenteropancreatic antigens in two cases of primary retroperitoneal mucinous cystoadenocarcinomas (PRMC) show that these tumours have patterns similar to ovarian mucinous tumours. Markers of pyloric type gastric mucosa differentiation (M1, cathepsin E, concavavalin A, pepsinogen II) are mostly positive in benign and borderline areas with endocervical type differentiation, while immunoreactivity for intestinal cell markers (M3SI and CAR-5) and for DU-PAN-2 is present mainly in frankly malignant areas, regardless of differentiation type. DNA analysis shows a point mutation of K-ras oncogene at codon 12 (GGT to CGT) in one case. The immunohistochemical and genotypic similarity of PRMC and ovarian mucinous tumours may indicate similar mechanisms in their histogenesis.
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  • 15
    Electronic Resource
    Electronic Resource
    Springer
    Virchows Archiv 424 (1994), S. 39-46 
    ISSN: 1432-2307
    Keywords: Proliferating cell nuclear antigen ; Immunohistochemistry ; Breast carcinomas
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract 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.
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  • 16
    ISSN: 1432-2307
    Keywords: v-src oncogene ; Rous sarcoma virus ; Fibrohistiocytic tumour ; Immunohistochemistry ; Southern blotting
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The observation that v-src-induced tumors contain tumor cells of differing morphology, notably fibroblastoid or polygonal, raised the question as to whether the tumor cells are also heterogeneous with respect to expression of markers of cellular differentiation. Of the markers tested here, consistent reactivity for tumor tissue was noted only for antibody probes reactive to muscle actin (HHF35, αsm-1) or to procollagen type I (SP1. D8); for any given tumor, whether induced by v-src DNA or by Rous sarcoma virus, each of these markers was found only in a subpopulation of tumor cells. The observation of marker heterogeneity in the one v-src DNA-induced tumor examined here that typed as monoclonal suggests that v-src-induced transformation is consonant with a degree of plasticity in the phenotypes of the clonal progeny of a single transformant.
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  • 17
    ISSN: 1432-2307
    Keywords: Gastric cancer ; Alpha-catenin ; Immunohistochemistry ; E-cadherin ; Cancer invasion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract E-cadherin (E-cad) plays a major role in the maintenance of cell-cell adhesion in epithelial tissues, and impaired E-cad expression correlates with tumour invasion and metastasis. Alpha-catenin (α-cat), an undercoat protein of adherens junctions, binds to the cytoplasmic domain of E-cad and is essential for linking E-cad to actin-based cytoskeleton. We investigated E-cad and α-cat expression in 60 human gastric cancers immunohistochemically. The 60 gastric cancers were classified into 18 (30%) in which α-cat expression was preserved, and 42 (70%) reduced cases. The reduction of α-cat expression was significantly related to dedifferentiation, depth of invasion, infiltrative growth and lymph node metastasis. We also examined the co-expression of α-cat and E-cad. Seventeen (28%) tumours preserved both molecules [α-cat(+)/E-cad(+)] and 33 (55%) tumours reduced both [α-cat(−)/E-cad(−)], whereas 9 (15%) tumours exhibited α-cat(−)/E-cad(+). The frequency of lymph node metastasis in α-cat(−)/E-cad(+) tumour (67%) was significantly higher than that in α-cat(+)/E-cad(+) tumours (24%) and was close to that in α-cat(−)/E-cad(−) tumours (82%). The frequency of haematogenous liver metastasis in α-cat(−)/E-cad(+) tumours (44%) was significantly higher than that in α-cat(+)/E-cad(+) tumours (6%) or α-cat(−)/E-cad(−) tumours (9%). Thus, in all E-cad(+) tumours, the frequency of lymph node and liver metastasis was higher in α-cat(−) tumours than in α-cat(+) tumours. α-Cat expression is apparently better at predicting tumour invasion and metastasis than E-cad expression.
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  • 18
    ISSN: 1432-2307
    Keywords: Cytokeratin Acute lymphoblastic leukaemia ; lymphoma ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract This report presents a case of common acute lymphoblastic leukaemia-lymphoma expressing low molecular weight cytokeratin but no leukocyte common antigen (CD45) in a 57-year-old man. The unusual morphology and clinical course together with the aberrant immunohistochemical results suggested a diagnosis of undifferentiated carcinoma. A detailed immunohistochemistry study on frozen and paraffin sections and molecular analysis prevented a diagnostic mistake.
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  • 19
    Electronic Resource
    Electronic Resource
    Springer
    Virchows Archiv 425 (1994), S. 69-72 
    ISSN: 1432-2307
    Keywords: GLUT-1 Glucose transporter protein ; Immunohistochemistry ; Microvessel endothelium ; Hippocampus ; Alzheimer's disease
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Changes in cerebral microvessel ultrastructure have been reported to occur in Alzheimer's disease (AD). In order to investigate whether these changes are associated with compromised blood-brain transport mechanisms, hippocampal formation sections from AD and age-matched normal brains were immunolabelled with an antibody to the GLUT-1 glucose transporter protein. GLUT-1 immunolabelling of microvessel endothelium was significantly reduced in the AD compared to normal hippocampal formation. Thus, AD is associated with a reduction in cerebral microvessel endothelium glucose transporter content, which may result in decreased glucose availability to the brain.
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  • 20
    ISSN: 1432-2307
    Keywords: Tenascin ; Stomach ; Hyperplasia Carcinoma ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Wie studied the expression of tenascin (Tn) in human stomach. In the normal mucosa of the antrum and body, Tn reaction was only seen in the muscularis mucosae, in the region of the pyloric sphincter and in the duodenum, a Tn-immunoreactive rim was seen underlying surface epithelial cells. Antral gastritis, irrespective of the degree of inflammation, showed a rim-like Tn expression under the surface epithelial cells but no Tn reaction was seen in mild chronic gastritis of the body. In some moderate and severe examples of chronic gastritis a delicate Tn-reactive line was seen to underline the surface epithelium focally and the neck regions of gastric pits. Discontinuous Tn immunoreactivity was sometimes seen beneath hyperplastic epithelium in both parts of the stomach. A Tn-immunoreactive line was seldom seen surrounding glands showing intestinal metaplasia. In both benign and malignant ulcers prominent Tn immunoreaction was seen at the base of ulcers extending deep into the underlying muscularis. Only severely dysplastic lesions displayed Tn in the lamina propria, in close association with capillaries. In early forms of diffuse gastric cancer (DGCA) raggedly increased Tn staining was seen in the lamina propria underlying affected surface epithelial cells. In advanced forms of DGCA consistent Tn expression was seen in the tumour stroma. A distinct Tn reaction was seen surrounding invasive tumour cell nests of intestinal type gastric cancer (IGCA) in the submucosa, whereas in early forms of the tumour enhanced Tn reaction was noted predominantly in the upper part of the lamina propria in the vicinity of dysplastic elements. Notably, while most invading DGCA tumour cell nests showed no Tn in the submucosa and muscle cell layer, invading IGCA islets showed prominent expression of Tn. The most conspicuous Tn enhancement in the stomach is seen in invasive tumours and in ulcers suggesting that Tn is an important stromal component in malignant growth and in lesions undergoing active repair and remodelling.
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  • 21
    ISSN: 1432-2307
    Keywords: Adenocarcinoma ; Immunohistochemistry ; Tumour suppressor gene ; Ethnicity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Gastric cancer is more than twice as common in Hispanics as in Anglos in Texas, while colorectal cancer is almost twice as common in Anglos as Hispanics. To test the hypothesis that mutations in the p53 tumour suppressor gene are involved in these differences, we examined 131 gastric and 138 colorectal cancers from Hispanic and Anglo patients from South Texas and Mexico using immunohistochemistry (IHC) as a screening assay for p53 mutations. The fraction of p53 positive cases was not significantly different in gastric cancers from Hispanics compared to Anglos (43% versus 61%, respectively, p=0.13) or in colorectal cancer (57% versus 58%, respectively, p=1.0), suggesting that p53 mutations are not involved in causing the different incidences of these cancers in these populations. In addition, the types of p53 mutations arising in gastric tumours from Hispanic patients were consistent with those reported in gastric tumours in other populations. Sequencing of mutations in five gastric cancers revealed two G: C to A: T transitions, two A: T to G: C transitions and one complex deletion. In contrast with findings in studies in other tumour types, neither stage nor survival was associated with p53 positive staining by IHC in either gastric or colorectal tumours in this study. Positive p53 immunostaining was associated with the diffuse histological subtype in gastric carcinoma (p=0.05) and high histological grade in colorectal carcinoma (p=0.04).
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  • 22
    ISSN: 1432-2307
    Keywords: C-kit product ; Immunohistochemistry ; Human normal tissue ; Small cell lung carcinoma ; Seminoma/dysgerminoma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Eighteen hundred and eighty-four cases of human solid tumours and 833 samples of normal human tissues, formalin-fixed and paraffin-embedded, were examined immunohistochemically for expression of c-kit oncogene product using polyclonal antibody against synthesized c-kit peptide. Seminoma/dysgerminoma and small cell lung carcinoma (SCLC) show preferential c-kit expression at 92% and 36% frequency, respectively, whereas only sporadic cases of cervical carcinoma and non-SCLC lung carcinoma show c-kit positivity. A normal tissue counterpart positive for c-kit product is detected in the testis (spermatocyte) and ovary (oocyte) but not in the lung or the cervix. In contrast, normal epithelial cells of the breast, skin basal cells and tissue mast cells harbour c-kit product, but transformed cells of the former two are largely deficient in the c-kit protein. One hundred and thirty-nine neuroendocrine tumours and 39 non-pulmonary small cell carcinomas were all negative, except for two cases of neuroblastoma. This indicates a distinct character for SCLC in c-kit expression. The c-kit product may be a useful marker in diagnostic pathology of seminoma/dysgermona and SCLC among human solid tumours, and in distinction of SCLC from non-pulmonary small cell carcinoma.
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  • 23
    ISSN: 1432-2307
    Keywords: p53 ; Immunohistochemistry ; Breast cancer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract To clarify whether p53 protein expression is involved in multistep carcinogenesis or the progression of mammary ductal carcinoma, we investigated p53 protein expression in 83 invasive ductal carcinomas (IDC), 10 IDC with a predominant intraductal component, 13 non-invasive ductal carcinoma (NIDC), 16 atypical ductal hyperplasia (ADH) and 39 benign epithelial hyperplasia (EH), using immunohistochemistry. Expression of p53 protein was detected in 24 (28.9%) cases of IDC, 5 (50%) cases of IDC with a predominant intraductal component and 1 (7.6%) case of NIDC. No expression was observed in either ADH or EH. In IDC, including cases with a predominant intraductal component, p53 protein expression was associated with a higher histological grade (P〈0.0001) or mitotic index (P〈0.0005). Although overexpression of c-erbB-2 protein has also shown a similar association with these prognostic indicators, expression of p53 protein correlated regardless of the status of c-erbB-2 overexpression. Completely coordinated expression of p53 protein was seen in both intraductal and invasive components. The intraductal component in IDC including cases with a predominant intraductal component which expresses p53 protein had significantly higher histological grade (P〈0.0005) or more comedo-subtypes (P〈0.0001). These results suggested that p53 protein expression occurs at a stage of NIDC with high histological grade or in comedo-subtypes. Its expression is maintained throughout invasion.
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  • 24
    ISSN: 1432-2307
    Keywords: Mesenchymoma ; Osteoclast ; Giant cell Malignant fibrous histiocytoma ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Three cases of malignant mesenchymoma with numerous osteoclast-like giant cells, arising in deep soft tissue, and which mimicked the so-called giant cell variant of “malignant fibrous histiocytoma” have been studied. All three neoplasms arose in adults; two patients were male and one was female. Two tumours arose in the thigh, and one in the right shoulder. Two patients died within 2 years of the primary excision while the third is alive and well at 2.5 years. Histologically, one case showed leiomyosarcoma plus liposarcoma, one leiomyosarcoma plus osteosarcoma, and one tumour consisted of liposarcoma plus osteosarcoma; all components were assessed morphologically as high-grade malignant. All three cases showed prominent osteoclast-like giant cells in the leiomyosarcomatous or osteosarcomatous areas, thereby closely mimicking the phenotype of so-called giant cell variant of “malignant fibrous histiocytoma”. We discuss briefly differences in soft tissue sarcomas demonstrating this distinctive osteoclast-rich phenotype.
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  • 25
    ISSN: 1432-2307
    Keywords: Thyroid ; Follicular tumour ; Oxyphilic cell tumour ; PCNA ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The expression of proliferative cell nuclear antigen (PCNA) in follicular tumours of the thyroid was examined by immunohistochemistry. Both usual nonoxyphilic cell follicular tumours (non-OCT) and oxyphilic cell tumours (OCT) were subdivided into benign, indeterminate, encapsulated carcinoma, and widely invasive carcinoma types. Among non-OCT the percentages of PCNA-positive cells in benign tumours, encapsulated carcinomas, and widely invasive carcinomas was 2.5%–8.6%, 11.8%–39.1%, and 18.6%–20.0%, respectively. There was a statistically significant difference between benign tumours and encapsulated or widely invasive carcinomas, as in previous studies. A value of 10% was appropriate to distinguish benign from malignant lesions. PCNA-positive cells in indeterminate-type non-OCT were not significantly different from those in benign tumours, ranging from 4.3%–19.6%, and occurring at more than 10% in three of six tuours. Among OCT the positivity was less than 10% in benign tumours (4.5%–7.8%) and more than 10% in malignant tumours (14.1%–35.9%) and all the eight indeterminate tumours (12.5%–27.3%), with a statistically significant differences between the benign tumour and each of the latter types. These results indicate that the examination of PCNA is valuable in diagnosis of thyroid follicular tumours and that the use of similar diagnostic criteria may be warranted in both non-OCT and OCT.
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  • 26
    ISSN: 1432-2307
    Keywords: Breast ; Fibrous histiocytoma ; Giant cells ; Flow cytometry ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We describe a benign mammary mesenchymal tumour with atypical stromal giant cells in the contralateral breast of a 66-year-old woman with infiltrating ductal carcinoma. The clinical, morphological and immunohistochemical features of this tumour suggest a pleomorphic variant of fibrous histiocytoma. This benign lesion represents a possible pitfall in breast pathology when interpreting a frozen section or fine needle aspiration biopsy.
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  • 27
    ISSN: 1432-2307
    Keywords: Intermediate filament proteins ; Cervix ; Neoplasia ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We investigated the expression of keratin subtypes 7, 8, 10, 13, 14, 17, 18 and 19 in the normal cervix, in cervical intraepithelial neoplasia (CIN) lesions and in cervical carcinomas, using a selected panel of monoclonal keratin antibodies, reactive with routinely processed, formalin fixed paraffin embedded tissue fragments. The reaction patterns derived for each keratin antibody were compared with known expression patterns of the various epithelia, previously examined in frozen tissues. Although the reactivity of the antibodies was generally acceptable, considerable modifications to the manufacturers' staining instructions were often necessary. For some antibodies, which were previously thought to be reactive with fresh frozen tissue only, we developed staining protocols rendering them reactive with routinely processed material. As with previous findings in frozen sections we observed increasing expression of keratins 7, 8, 17, 18 and 19 with increasing grade of CIN. In cervical carcinomas the differences in keratin detectability between the main categories were more pronounced than in frozen sections, probably due to fixation and processing. For routine pathology, keratin phenotyping of cervical lesions may be of value in classification. The fact that keratin 7 was detected for the first time in reserve cells, and that this keratin was also found to be expressed in a considerable number of CIN lesions and cervical carcinomas supports the suggestion that reserve cells are a common progenitor cell for these lesions.
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  • 28
    ISSN: 1432-2307
    Keywords: Metallothionein ; Immunohistochemistry ; Colorectal cancer ; Prognosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Metallothioneins (MTs), a set of ubiquitous low-molecular-weight proteins essential for the protection of cells against heavy metal ion toxicity, were demonstrated immunohistochemically using a monoclonal antibody (E9) against a conserved epitope of I and II isoforms in a series of 109 colorectal adenocarcinomas. In a semiquantitative analysis strong MT expression in the majority of tumour cells was observed in 34 (31%) cases, 24 (22%) tumours showed a focal MT positivity, and 51 (47%) almost completely lacked MT expression. These differences in MT expression were statistically significantly (P〈0.05) associated with the tumour stage (Dukes classification) and the lymph node involvement at the time of operation (pN stages). However, in contrast to previous findings obtained on a variety of tumours, MT positivity was associated with a favourable clinical outcome in colonic carcinoma, which may indicate their different biological behaviour. Survival curves of cases with MT-positive and MT-negative status differed from each other in a univariate analysis (Mantel-Haenszel: 8.9, P〈0.05) but lost significance when a multivariate analysis was carried out by means of the Cox proportional regression model with Dukes' stages as a stratification factor. It is concluded that immunohistochemically demonstrated MT expression is significantly associated with tumour stages but does not represent an independent prognostic variable in colorectal cancer. However, it may provide important information about some of the biological mechanisms underlying progression in colorectal cancer.
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  • 29
    ISSN: 1432-1920
    Keywords: Pituitary adenoma ; MRI ; Immunohistochemistry ; Pituitary hormones
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Our aim was to elucidate the factors which determine the MRI signal intensities of pituitary adenomas. We examined 51 patients with surgically-confirmed pituitary adenomas. Using a spin-echo pulse sequence (SE 500/15), coronal and sagittal images (3 mm slices) were obtained. Signal intensities on T1-weighted images were measured in the parenchyma of the adenoma and in normal grey matter. The relative intensity of the adenoma was assessed by calculating the ratio of its signal intensity to that of the normal grey matter of the same patient. Parafin-embedded sections were used for haematoxylin and eosin staining. The number of cells in a prescribed area was counted, and the mean of five such counts was taken as the cell density. Immunohistochemically stained sections using antibodies for various pituitary hormones were similarly examined; the ratio of the total number of hormone-positive cells to the overall total number of adenoma cells was calculated. Four independent variables were used in the analysis: the age of the patient, the maximum diameter of the adenoma, the cell density and the proportion of hormone-positive cells in the adenoma and, with the signal intensity ratio as the dependent variable, a multiple regression analysis was performed. This revealed that the the greatest influence upon the signal intensities on T1-weighted images was the proportion of hormone positive cells.
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  • 30
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    Der Pathologe 15 (1994), S. 181-186 
    ISSN: 1432-1963
    Keywords: Schlüsselwörter Angiosarkom ; Nebenniere ; Immunhistochemie ; Intermediärfilamente ; Zytokeratine ; CD 31 ; Key words Angiosarcoma ; Adrenal gland ; Immunohistochemistry ; Intermediate filaments ; Cytokeratins ; CD 31
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Cytological, biopsy and autopsy findings in a patient suffering from massively metastasizing adrenal angiosarcoma are reported. Histogenetic typing of the tumour initially manifestating itself by osseous and liver metastases was problematic with regard to its partially epithelioid structure and its positivity upon cytokeratin immunostaining. Of relevance for the correct typing was the finding that the tumour cells in addition exhibited positivity for vascular markers. This case confirms literature data according to which cytokeratin expression not infrequently may be encountered in endothelial neoplasms and which by no means should lead to exclude such a tentative diagnosis.
    Notes: Zusammenfassung Berichtet wird über zytologische, bioptische und autoptische Befunde bei einem Patienten mit fortgeschritten metastasiertem adrenalen Angiosarkom. Die korrekte histogenetische Klassifikation der sich zunächst durch ossäre und Leberabsiedlungen manifestierenden teilweise epitheloiden Neoplasie war konventionell-morphologisch und wegen des immunhistologisch positiven Zytokeratin-Nachweises problematisch. Bedeutung für die Diagnosesicherung hatte die zusätzlich nachgewiesene Positivität der Tumorzellen für vaskuläre Marker. Der beschriebene Fall bestätigt Literaturbefunde, nach denen eine Zytokeratin-Expression unter Angiosarkomen nicht selten angetroffen wird und keinesfalls zum Ausschluß einer entsprechenden Verdachtsdiagnose führen darf.
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  • 31
    ISSN: 1432-1963
    Keywords: Schlüsselwörter Autoklavieren ; Mikrowelle ; Immunhistochemie ; Antigen-Demaskierung ; Key words Wet autoclaving ; Microwave pretreatment ; Immunohistochemistry ; Antigen retrieval
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Wet autoclaving is a simple, reliable and time-effective method for antigen retrieval in routinely processed archival material. Both routine diagnostic (e. g., oestrogen and progesterone receptors, cytoskeletal proteins) and research antibodies (e. g. various p53 antibodies, mdm-2, bcl-2, MIB-1) are reported to demonstrate its application. Wet autoclaving may allow successful application of antibodies in paraffin-embedded tissues designed for use on frozen sections. The technique has the poten-tial to reliably handle up to 200 sections at a time, without evidence of any significant damage to the sections or nuclear morphology.
    Notes: Zusammenfassung Mit der Technik des feuchten Autoklavierens wird eine einfache, verläßliche und zeitsparende Methode zur Antigen-Demaskierung an Formalin-fixiertem und Paraffin-eingebettetem Gewebe vorgestellt. Anhand einer Reihe von Antikörpern (Östrogen- und Progesteronrezeptoren, Zytoskelettproteine, verschiedene p53-Antikörper, mdm-2, bcl-2, MIB-1 u. a.) verwendeter Antikörper werden die Vorteile dieser Methode beschrieben. Das feuchte Autoklavieren ermöglicht bei einigen sonst nur am Gefrierschnitt einsetzbaren Antikörpern auch deren Anwendung am Paraffinschnitt. Für den Routinepathologen ist die leichte Handhabung sowie die hohe Reproduzierbarkeit von Vorteil.
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  • 32
    ISSN: 1432-1963
    Keywords: Schlüsselwörter bcl-2 ; Osteosarkom ; Apoptose ; programmierter Zelltod ; Immunhistologie ; Proliferation ; Key words bcl-2 ; Osteosarcoma ; Apoptosis ; programmed cell death ; Immunohistochemistry ; Proliferation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary The relationsship between the growth of tumors and the expression of the protooncogen Bcl-2 could be shown in epithelial tumors. A bcl-2 expression leads to a prolonged cell survival due to an inhibition of apoptosis. The potential meaning of bcl-2 expression in mesenchymal tumors remains still unknown. The fact, that the heterogenous group of osteosarkoma is not sufficiently characterized at present, suggested to investigate the bcl-2 expression in osteosarcoma. Thus, immunohistochemistry was used to analyze 47 specimens of different osteosarcomas of 36 patients. Sixteen cases (46 %) showed a strong expression of bcl-2 and 13 cases (35 %) were moderately positiv for bcl-2. Seven cases (19 %) were negative for bcl-2. The heterogenous, negative up to strong expression of bcl-2 yield clues, that the Bcl-2 controlled regulation of programmed cell death could be an important factor of cellular kinetics. Additionally the cellular proliferationrate was determined with the monoklonal antibody MIB 1, directed against the Ki-67 epitop. The data of bcl-2 expression and cellular proliferationrate lead to a classification correlating with the histological classification. To verify the importance of apoptosis in the genesis of mesenchymal tumors and whether Bcl-2 may play an important role as a predictive factor for the prognosis of osteosarcoma, further investigations will be needed.
    Notes: Zusammenfassung Bei zahlreichen epithelialen Geweben konnte ein Zusammenhang zwischen Tumorwachstum und der Expression des Protoonkogens Bcl-2 nachgewiesen werden. Eine bcl-2-Expression ist verbunden mit verlängertem Zellüberleben infolge einer Apoptoseinhibition. Hingegen ist über die bcl-2-Expression und deren mögliche Bedeutung in mesenchymalen Tumoren wenig bekannt. Da die heterogene Gruppe der Osteosarkome mit den derzeitigen methodischen Mitteln nicht hinreichend charakterisierbar ist, wurde die bcl-2-Expression untersucht. Immunhistologisch wurden 47 Osteosarkompräparate von 36 Patienten unterschiedlicher Subtypen analysiert. Von den 36 Fällen zeigten in der Biopsie 16 Fälle (46 %) eine stark positive und 13 Fälle (35 %) eine mittelgradig positive bcl-2 Expression. Sieben Fälle (19 %) waren bcl-2-negativ. Die heterogene, fehlende bis starke bcl-2-Expression deutet darauf hin, daß in Osteosarkomen die Bcl-2-gesteuerte Regulation des programmierten Zelltodes einen Faktor in der zellulären Wachstumskinetik darstellt. Zusätzlich wurde die Proliferationsrate, anhand des gegen das Ki-67-Antigen gerichteten monoklonalen Antikörper MIB-1 bestimmt. Aus den Daten zur bcl-2-Expression und Proliferationsrate ergibt sich eine Einteilung, die eine Übereinstimmung mit der histologischen Klassifikation aufweist. Welche Bedeutung die Apoptose in der Genese mesenchymaler Tumoren hat und ob die bcl-2-Expression einen prädiktiven Wert für die Prognose von Osteosarkomen besitzt, bedarf weiterer Untersuchungen.
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  • 33
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    Der Pathologe 15 (1994), S. 358-360 
    ISSN: 1432-1963
    Keywords: Schlüsselwörter Tonsillentumor ; Primär extrakranielles Meningeom ; Atypisches Meningeom ; Immunhistologie ; Key words Tumour of the palatine tonsil ; Primary extracranial meningioma ; Atypical meningioma ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary To our knowledge this is the first reported case of a primary extracranial meningioma located in the palatine tonsil. Immunohistochemical investigation of the tumour showed coexpression of vimentin and neuron-specific enolase (NSE). No staining was found with antibodies against cytokeratins KL 1, 13/10, 8 and 18, epithelial membrane antigen EMA and melanoma protein (HMB-45). It seems justifiable to classify this tumour as an atypical meningioma because of the local increased mitotic activity.
    Notes: Zusammenfassung Dieses ist nach unseren Kenntnissen der 1. Fallbericht eines primär extrakraniellen, in der Tonsille lokalisierten Meningeoms. Immunhistologisch wies der Tumor eine Koexpression von Vimentin und neuronspezifischer Enolase (NSE) und eine negative Reaktion mit Antikörpern gegen die Zytokeratine KL 1, 13/10, 8, 18, das epitheliale Membranantigen EMA und Melanom Protein (HMB 45) auf. Aufgrund der lokal gesteigerten Mitoserate scheint es gerechtfertigt, den vorliegenden Tumor als atypisches Meningeom zu klassifizieren.
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  • 34
    ISSN: 1432-0827
    Keywords: Decorin ; Proteoglycan-100 ; Heterotopic ossification ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Heterotopic ossification is a metabolically active process which shares several properties of orthotopic bone formation and, therefore, represents an excellent model for studying bone matrix components. Immunohistochemical methods were used to investigate the distribution pattern of the small proteoglycans decorin and proteoglycan-100 during different stages of heterotopic ossification of pressure sores of paraplegic patients. Decorin and proteoglycan-100 exhibited a substantially divergent distribution pattern. Decorin was detectable in the perivascular matrix of granulation tissue as well as in the stroma of heterotopic ossification. The ossification zone was stained most strongly. In contrast, proteoglycan-100 was predominantly detectable in fibroblasts and preosteoblasts in early areas of osteogenesis. In more mature forms of heterotopic ossification immunostaining was markedly reduced in osteoblasts and osteocytes and even absent in so-called bone-lining cells. However, at least some osteoclasts were strongly positive. These results suggest indicate that decorin and proteoglycan-100 are important components during the formal pathogenesis of heterotopic ossification. The expression of the small proteoglycans, especially of proteoglycan-100, correlates with different phases during heterotopic ossification, showing a maximum for proteoglycan-100 in matrix-forming cells in early phases of bone formation, but in osteoclasts in mature bone.
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  • 35
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    Calcified tissue international 55 (1994), S. 38-45 
    ISSN: 1432-0827
    Keywords: Vacuolar-type H+-ATPase ; Carbonic anhydrase II ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract The localization of vacuolar-type H+-ATPase and carbonic anhydrase II (CA II) in rat incisor enamel organs at maturation was examined by light and electron microscopy. The immunoreactivity for both vacuolar-type H+-ATPase and CA II was intense on the ruffled border of ruffle-ended ameloblasts (RA), but moderate at the distal end of smooth-ended ameloblasts (SA). Immuno-gold particles indicated that CA II was not confined to the ruffled border of RA alone, but also distributed in the cytoplasm of RA and SA. These findings suggest that RA may secrete protons produced by CA II via the ruffled border into enamel by active transport of vacuolar-type H+-ATPase. Secreted protons may activate hydrolytic enzymes to degrade the organic components of enamel matrix. Vacuolar-type H+-ATPase on vesicles of SA suggests that a specific configuration of ruffled borders in RA may be formed by the fusion of vesicle membranes in the distal end of cytoplasm of SA.
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  • 36
    ISSN: 1432-0738
    Keywords: Key words Nasal cavity ; Immunohistochemistry ; Glutathione S-transferases
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  Many xenobiotics induce lesions within the nasal cavity of experimental animals which are site specific. This site selectivity may be due to regional deposition within the nasal cavity and/or the localisation of biotransformation enzymes. We have developed methodology which allows immunohistochemical localisation of xenobiotic biotransformation enzymes in transverse sections of the rat nasal cavity identical to those normally taken for pathological examination. We report the application of this methodology to six isoenzymes of the glutathione S-transferases (GSTs). All six isoenzymes were predominantly located within olfactory epithelium covering the ethmoturbinates (levels III and IV) and extending forwards into the dorsal meatus (level II). Squamous and transitional epithelia showed little or no staining while respiratory epithelium was weakly stained. Within the respiratory epithelium only the ciliated columnar cells and, to a lesser extent, some of the seromucous glands contained GSTs. Within olfactory epithelium the sustentacular cells, basal cells and subepithelial glands all stained positive for GSTs. The different cell types of olfactory epithelium preferentially express different GST isoenzymes: 1-1 and 2-2 were predominantly located in the subepithelial glands; 3-3, 4-4 and 8-8 in sustentacular and basal cells; 7-7 in basal cells.
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  • 37
    ISSN: 1432-0533
    Keywords: Managanese superoxide dismutase ; Amyotrophic lateral sclerosis ; Free radical ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Neuronal expression of manganese superoxide dismutase (MnSOD) in sporadic amyotrophic lateral sclerosis (sALS) was investigated by an immunohistochemical method. The brains and spinal cords from 11 patients with sALS and 20 normal controls (NCs) were used, and the following four nuclei (three motor nuclei and one autonomic nucleus) were examined: the oculomotor nucleus; the hypoglossal nucleus; the cervical motor nucleus; and Onuf's nucleus. Serial sections were stained by the Klüver-Barrera (KB) method and with human-MnSOD-specific antibodies. We counted the total number of neurons visible after KB staining and the total number of positive neurons after immunostaining. The average total number of neurons after KB staining was similar in sALS patients and NCs in both the oculomotor nucleus and Onuf's nucleus, but the number in the hypoglossal and cervical motor nuclei was significantly lower in sALS. The ratio of MnSOD-positive neurons to total neurons visible after KB staining, calculated as an index of the expression of MnSOD, was significantly higher in the oculomotor nucleus and Onuf's nucleus, and lower in the hypoglossal nucleus in sALS patients than in NCs. In the cervical motor nucleus, the ratio in sALS patients did not differ from that in NCs. These results suggest that production of toxic superoxide radicals might be increased in sALS, and that neurons that successfully induce the expression of sufficient MnSOD can survive the disease process, while those failing to activate adequate expression of the enzyme succumb to the toxic effects of the radicals and die.
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  • 38
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    Acta neuropathologica 87 (1994), S. 398-404 
    ISSN: 1432-0533
    Keywords: Astrocyte cultures ; Brain stem infection ; Herpes simplex virus type 1 ; Immunohistochemistry ; Rat model
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Infection of the CNS by herpes simplex type 1 (HSV-1) via the trigeminal route to the brain stem was elucidated in a rat model. In contrast to the earlier described cortical and hippocampal infection after intracranial injection, the CNS showed a profound resistance to HSV-1 infection when the virus was administred by nose inoculation, as judged by histopathology and immunohistochemistry. In contrast, when the distribution of HSV-1 in the brain was investigated after nose inoculation by polymerase chain reaction, viral DNA was detected at all levels from the ganglia to the cortex. When replication of HSV-1 was assayed in primary cell cultures of rat astrocytes derived from brain stem, striatum, hippocampus and cerebral cortex, significantly lower virus yields were obtained in brain stem-derived astrocytes cultures as compared with in cortex-derived astrocytes. This finding was independent of whether HSV-1 strains used originated from brains of patients suffering from herpes simplex encephalitis or from patients with oral cutaneous lesions and lacking neurological symptoms. Also, by immunocytochemistry of cultures after HSV-1 infection, a lower number of plaques were seen in brain stem-derived astrocytes as compared with cortex-derived astrocytes. The observed relative resistance of brain stem-derived astrocytes to replicate HSV-1 might contribute to the ability of the brain stem to withstand infection during reactivation of this virus in the trigeminal neurons.
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  • 39
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    Acta neuropathologica 88 (1994), S. 454-458 
    ISSN: 1432-0533
    Keywords: Immunohistochemistry ; Meningioma ; Neurofibroma ; Schwannoma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract CD34 is a sialylated transmembrane glyco-protein of unknown function that is present in myeloid progenitor cells, endothelial cells, and some fibroblastrelated mesenchymal cells. However, its tissue distribution is still incompletely characterized. In this study we evaluated the distribution of CD34 antigen in tumors of the central and peripheral nervous system. For comparison the tumors were also stained for CD31, also known as platelet-endothelium cell adhesion molecule (PECAM-1), a transmembrane glycoprotein so far considered to be endothelium specific beyond its reactivity with certain hematopoietic cells. Neurofibromas showed consistently high numbers of CD34-positive spindle cells, whereas peripheral and acoustic schwannomas were negative. A subset of meningiomas (15%) showed CD34-positive tumor cells, and some were also weakly positive for CD31. Gliomas were negative. Meningeal hemangiopericytomas were consistently CD34 positive, but CD31 negative. These results indicate a moderately widespread distribution of the CD34 antigen in nervous system tumors, and necessitate caution in making conclusions regarding endothelial cell differentiation of nervous system tumors on the basis of CD34 immunoreactivity.
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  • 40
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    Anatomy and embryology 189 (1994), S. 237-242 
    ISSN: 1432-0568
    Keywords: Thyroid cartilage ; Immunohistochemistry ; Mineralization ; Ossification ; Collagens
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Thyroid cartilages of various ages were investigated by immunofluorescence staining for localization of the fibrillar collagen types I and II in order to understand the tissue remodeling occurring during the mineralization and ossification of thyroid cartilage. In fetal and juvenile thyroid cartilages, type I collagen was restricted to the inner and outer perichondrium, while type II collagen was localized in the matrix of hyaline cartilage. However, in advanced ages, type I collagen was also localized in the pericellular and in the interterritorial matrix of intermediate and central chondrocytes of thyroid cartilage. The matrix of peripheral chondrocytes was negative for type I collagen. This suggests that some chondrocytes in thyroid cartilage undergo a differentiation to type I collagen-producing chondrocytes. At the beginning of ossification, bone-related type I collagen was chiefly detected in the central cartilage layer, but was never deposited first from the perichondrium in the direction to the subperichondrial cartilage. This observation confirmed previous findings showing that osteogenesis mainly follows an endochondral ossification pattern. Interterritorial matrix failed to react with the type II collagen antibody in men from the beginning of the third decade, and later still in women, even after treatment with hyaluronidase. These observations indicate that major matrix changes occur faster in male than in female thyroid cartilage.
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  • 41
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    Archives of dermatological research 286 (1994), S. 62-68 
    ISSN: 1432-069X
    Keywords: Metallothionein ; Immunohistochemistry ; Monoclonal antibodies
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The expression and distribution of metallothionein (MT) in frozen sections of normal and pathological human skin was studied using the monoclonal antibody L2E3 directed against MT derived from human fetal liver. Immunohistochemical staining of normal fetal and adult skin revealed strong reactivity in basal keratinocytes of epidermis and outer hair root sheath, hair matrix cells and the secretory coil, but not the exocrine portion of eccrine glands; myoepithelial cells around apocrine sweat glands were similarly stained. In epidermal hyperplasia, variable numbers of suprabasal keratinocytes were stained, whereas in interface dermatitis, interrupted staining was found in the basal layer. Weak or scattered staining was observed in squamous tumours, whereas basal cell carcinomas did not show consistent staining. The distribution of MT in normal skin was in line with the germinative role of basal keratinocytes and hair matrix cells, whereas its distribution in hyperplastic epidermis was in line with experimental animal data, and reflected the increase in the germinative pool in these conditions. It is concluded that monoclonal antibody L2E3 may serve as a valuable immunohistochemical marker in diagnostic cutaneous pathology since it labels basal keratinocytes selectively, and since it discriminates between eccrine and apocrine sweat glands.
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  • 42
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    Archives of dermatological research 287 (1994), S. 28-35 
    ISSN: 1432-069X
    Keywords: Melanoma pathology ; Antigen CD analysis ; Tumor-infiltrating lymphocytes ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Various clinical and experimental observations point to the existence of an immunological host defense in cutaneous malignant melanoma. To identify the major effector mechanisms mediating the specific anti-tumor immune response, we examined 23 benign and neoplastic melanocytic lesions (3 nevi, 14 primary melanomas, and 3 cutaneous and 3 systemic metastases) by quantitative immunohistology, and correlated these results with the histopathological and clinical subtypes of malignant melanoma. Our analyses indicate that CD3+ T-cell receptor α/Β-expressing lymphocytes are the prevailing leukocyte subset in primary as well as secondary malignant melanoma. We further observed that in early lesions (〈0.75 mm) of superficial spreading melanoma the vast majority of tumor-infiltrating lymphocytes (TIL) belong to the CD4+ subset and frequently express CD45RA antigens. In more advanced tumors, the contribution of CD8+ TIL gradually increases, indicating that the quality of the anti-tumor immune response changes during the course of the disease. Finally, we found that a varying percentage of cutaneous TIL express the cutaneous leukocyte antigen which is defined by the monoclonal antibody HECA 452 and preferentially expressed by skin-seeking memory T cells. In contrast, extracutaneous melanoma metastases (liver, brain, ovary) were completely devoid of HECA 452-reactive lymphocytes. These findings suggest that lymphocytes infiltrating cutaneous melanomas belong to a memory/effector T-cell subset functionally associated with the skin.
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  • 43
    ISSN: 1432-069X
    Keywords: Mixed tumour of skin ; Bone morphogenetic protein ; Chondrogenesis ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The distribution of immunoreactivity of bone morphogenetic protein (BMP), the glycosaminoglycans chondroitin 4-sulphate (C4SPG), chondroitin 6-sulphate (C6SPG), dermatan sulphate (DSPG) and keratan sulphate proteoglycans (KSPG), cytokeratin (K8.12), vimentin, glial fibrillary acidic protein (GFAP), actin, desmin, S-100 protein and neuron-specific enolase (NSE) in mixed tumour of the skin was investigated using immunohistochemical methods using monoclonal (MoAb) and polyclonal antibodies (PoAb). A strong BMP immunoreactivity was found characteristically in outer tumour cells of tubuloductal structures and modified myoepithelial cells. Modified myoepithelial cells and chondroidally changed cells showed positive immunoreactivity for C4SPG, C6SPG and DSPG; and KSPG was more pronounced in the modified myoepithelial cells. Vimentin, S-100 protein, GFAP and NSE, but not actin and desmin, were distribute in the outer tumour cells and modified myoepithelial cells in chondroidally changed tissue. Two factors show that chondrogenesis in mixed tumour of the skin is associated with the modified myoepithelial cells through the activity of BMP and biosynthesis of glycosaminoglycans as matrix substance. First, outer or basal tumour cells in mixed tumour of the skin is characterized by the presence of positive immunoreactivity for BMP, KSPG, vimentin, cytokeratin K8.12, S-100 protein, GFAP and NSE, and second, there is a matrix of chondroidally changed tissue containing the reaction products of C4SPG, C6SPG, DSPF and KSPG.
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  • 44
    ISSN: 1432-0533
    Keywords: Key words: Manganese superoxide dismutase ; Amyotrophic lateral sclerosis ; Free radical ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Neuronal expression of manganese superoxide dismutase (MnSOD) in sporadic amyotrophic lateral sclerosis (sALS) was investigated by an immunohistochemical method. The brains and spinal cords from 11 patients with sALS and 20 normal controls (NCs) were used, and the following four nuclei (three motor nuclei and one autonomic nucleus) were examined: the oculomotor nucleus; the hypoglossal nucleus; the cervical motor nucleus ; and Onuf's nucleus. Serial sections were stained by the Klüver-Barrera (KB) method and with human-MnSOD-specific antibodies. We counted the total number of neurons visible after KB staining and the total number of positive neurons after immunostaining. The average total number of neurons after KB staining was similar in sALS patients and NCs in both the oculomotor nucleus and Onuf's nucleus, but the number in the hypoglossal and cervical motor nuclei was significantly lower in sALS. The ratio of MnSOD-positive neurons to total neurons visible after KB staining, calculated as an index of the expression of MnSOD, was significantly higher in the oculomotor nucleus and Onuf's nucleus, and lower in the hypoglossal nucleus in sALS patients than in NCs. In the cervical motor nucleus, the ratio in sALS patients did not differ from that in NCs. These results suggest that production of toxic superoxide radicals might be increased in sALS, and that neurons that successfully induce the expression of sufficient MnSOD can survive the disease process, while those failing to activate adequate expression of the enzyme succumb to the toxic effects of the radicals and die.
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  • 45
    ISSN: 1432-0533
    Keywords: Key words: Perineurial cells ; Nerve regeneration ; Immunohistochemistry ; Epithelial membrane antigen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Perineurial cells are specialized connective tissue cells that form a barrier between endoneurium and epineurium in normal nerves. In the present study, the formation of the perineurium after transection of rat sciatic nerves was investigated. The cord bridging the gap between proximal and distal stumps through silicone tubes was studied 3, 7, 12, 18, and 21 days after surgery using electron microscopy and antibodies against epithelial membrane antigen (EMA), a marker for perineurial cells that has thus far not been applied to the study of differentiating cells in nerve tubulation systems. Initially, a thin cord consisting of fibrin bridged the gap between the stumps. At 7 days, longitudinal cells had migrated from both stumps toward the center of the tubes on the surface of the fibrin cord. These cells were immunoreactive with anti-EMA. At 12 days, ultrastructural features of perineurial cells (desmosomes, tight junctions, actin filaments with dense bodies, tonofilaments) were prominent in these cells. Subsequently, the gap was bridged through the perineurial tube by endothelial cells, pericytes, fibroblasts, Schwann cells, and axons. At 21 days, a single large nerve fascicle ensheathed by a mature perineurium was found between the stumps. Thus, the first cells to connect proximal and distal stumps in the investigated nerve regeneration silicon chamber system are perineurial cells. Through the tube formed by these cells, blood vessels and nerve fibers bridge the gap. Therefore, establishment of a perineurial connection between nerve stumps appears to be important in the sequence of events during nerve regeneration.
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  • 46
    ISSN: 1432-0533
    Keywords: Round granulated body ; Eosinophilic hyaline droplets ; Astrocytic tumors ; Immunohistochemistry ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Round granulated body (RGB) and eosinophilic hyaline droplets (EHDs) have been described as cytoplasmic inclusions of certain astrocytic tumors. In the previous literature, however, these inclusions have been described using various terms or regarded as nosologically the same entity. Light microscopically, RGB apeared as a round discrete body filled with fine uniform granules, while EHDs demonstrated a cluster of bright eosinophilic, round objects of various size. They could be clearly distinguished even by conventional histochemical staining such as the Masson trichrome stain and the phosphotungstic acid hematoxylin preparation. Both RGB and EHDs expressed positive immunoreactions for glial fibrillary acidic protein, several lysosomal markers, and some stress-response proteins. The ultrastructural appearances of these inclusions were distinct, however, one common feature was that they consisted of aggregations of numerous membrane-bound electron-dense bodies. Thus, both inclusions appear to be produced by neoplastic astrocytes and are possibly related to the lysosomal system. We examined the presence of RGB and EHDs in 138 astrocytic tumors. Both inclusions occurred most frequently in pleomorphic xanthoastrocytomas, followed by gangliogliomas and pilocytic astrocytomas. Subependymal giant cell astrocytomas exhibited only RGBs. RGBs and EHDs were not seen in any abundance in glioblastomas, gliosarcomas, fibrillary astrocytomas, protoplasmic astrocytomas, or oligo-astrocytomas. Some glioblastomas, however, showed only EHDs in small numbers. Several anaplastic astrocytomas were associated with a large number of RGBs and/or EHDs, and they revealed only rare mitosis despite marked cellular pleomorphism. Although RGB and EHDs have different morphological features, the presence of these inclusions in abundance may represent either a degenerative change, a long-standing lesion, or an indolent growth of the astrocytic tumors.
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  • 47
    ISSN: 1432-0533
    Keywords: Key words Round granulated body ; Eosinophilic ; hyaline droplets ; Astrocytic tumors ; Immunohistochemistry ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Round granulated body (RGB) and eosinophilic hyaline droplets (EHDs) have been described as cytoplasmic inclusions of certain astrocytic tumors. In the previous literature, however, these inclusions have been described using various terms or regarded as nosologically the same entity. Light microscopically, RGB appeared as a round discrete body filled with fine uniform granules, while EHDs demonstrated a cluster of bright eosinophilic, round objects of various size. They could be clearly distinguished even by conventional histochemical staining such as the Masson trichrome stain and the phosphotungstic acid hematoxylin preparation. Both RGB and EHDs expressed positive immunoreactions for glial fibrillary acidic protein, several lysosomal markers, and some stress-response proteins. The ultrastructural appearances of these inclusions were distinct, however, one common feature was that they consisted of aggregations of numerous membrane-bound electron-dense bodies. Thus, both inclusions appear to be produced by neoplastic astrocytes and are possibly related to the lysosomal system. We examined the presence of RGB and EHDs in 138 astrocytic tumors. Both inclusions occurred most frequently in pleomorphic xanthoastrocytomas, followed by gangliogliomas and pilocytic astrocytomas. Subependymal giant cell astrocytomas exhibited only RGBs. RGBs and EHDs were not seen in any abundance in glioblastomas, gliosarcomas, fibrillary astrocytomas, protoplasmic astrocytomas, or oligo-astrocytomas. Some glioblastomas, however, showed only EHDs in small numbers. Several anaplastic astrocytomas were associated with a large number of RGBs and/or EHDs, and they revealed only rare mitosis despite marked cellular pleomorphism. Although RGB and EHDs have different morphological features, the presence of these inclusions in abundance may represent either a degenerative change, a long-standing lesion, or an indolent growth of the astrocytic tumors.
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  • 48
    ISSN: 1432-0533
    Keywords: Immunohistochemistry ; Medulla oblongata ; Prion protein (PrP) ; Bovine spongiform encephalopathy ; Formalin-fixed tissue
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The efficacy of three pretreatment techniques for the detection of prion protein (PrP) in formalin-fixed, paraffin-embedded bovine spongiform encephalopathy (BSE)-affected brain tissue were compared using automated image analysis. The most abundant immunostaining was in the form of particulate expression observed in sections pretreated with hydrated autoclaving for 30 min. Considerably less immunostaining occurred in sections pretreated with formic acid and no specific particulate immunostaining was detected in sections pretreated with hydrolytic autoclaving. Hydrated autoclaving pretreatment of sections prior to PrP immunolabelling gives visualisation of widespread sites of abnormal PrP deposition in the brain, allowing detailed study of the form and distribution of the protein in routinely fixed bovine central nervous system affected with BSE.
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  • 49
    ISSN: 1432-0533
    Keywords: MyoD1 ; Myopathy ; Immunohistochemistry ; Neurogenic atrophy ; Werdnig-Hoffmann's disease
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The expression of the myogenic determination gene MyoD1 plays a primary role in the commitment of primitive mesenchymal cells to a striated muscle lineage and is down-regulated during later stages of differentiation. To determine the potential role of this gene in myopathic conditions, we examined its expression by means of immunohistochemical analysis, using a series of muscle biopsies from 14 patients with a variety of primary myopathies and neurogenic disorders. Utilizing the avidin-biotin-complex technique, cryostat sections were stained with monoclonal antibody 5.8 A, which we have previously described as having a high level of specificity for tumors with rhabdomyoblastic differentiation. Of special interest was the observation in 4 of 8 cases of neurogenic atrophy of varying levels of cytoplasmic positivity of muscle fibers, appearing to correlate with their degree of atrophy, in addition to weak nuclear staining. Muscle biopsies from 2 patients with Duchenne's muscular dystrophy and 2 patients with autoimmune inflammatory myopathies demonstrated various levels of nuclear positivity in scattered foci that appeared to correlate with areas of regeneration. A biopsy from a single case of neurogenic atrophy secondary to infantile spinal muscular atrophy (Werdnig-Hoffmann's disease) demonstrated diffuse but relatively weak staining of myofiber nuclei, in contrast to sections of normal striated muscle and muscle biopsies from patients with unexplained myoglobinuria, which exhibited only minimal amounts of staining. These data are compatible with observations that MyoD1 expression is related to electrical activity and muscle regeneration.
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  • 50
    ISSN: 1432-0533
    Keywords: Inclusion body disease ; Electron microscopy ; Immunohistochemistry ; Viral infection ; Primary metabolic disorder
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A Caucasian female who was noted to be mildly microcephalic at birth was diagnosed as having cerebral palsy at the age of 1 year. Her development was delayed and she never walked or talked. She appeared relatively stable neurologically until the age of 17 years when she had an illness with fever thought to be due to a virus. She was noted to deteriorate from this time on until her death at the age of 19 years. Autopsy revealed intranuclear and cytoplasmic inclusions widespread throughout the brain and visceral organs. There was no evidence of inflammation. Immunohistochemistry revealed strong immunoreactivity for tau protein and neurofilament protein. Electron microscopy revealed the inclusions to be composed of homogeneous finely granular material. Scattered with the granular material in the cytoplasmic bodies were crystalline structures with a honeycomb appearance. The possibility of these changes representing an old viral infection or a primary metabolic disorder are discussed.
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  • 51
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 190 (1994), S. 55-63 
    ISSN: 1432-0568
    Keywords: Neuropeptide Y ; Corpus callosum ; Development ; Transitory axons ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Many immunocytochemical studies have identified different types of neurotransmitters localized in the corpus callosum (CC) axons in the adult mammal. Few studies have looked at the development of different neurochemically identified CC systems. Previous studies on the development of cat CC axons have indicated that a large number of transitory CC axons project to the cortex during early postnatal development. The present study focuses on the development of one neurochemically identified group of CC axons in the cat, labeled with an antibody against neuropeptide Y (NPY), to determine if this group participates in transitory CC axonal growth. Cats at specified ages from birth to adulthood were studied with a routine method of immunocytochemistry for antiserum to NPY. NPY-immunoreactive (ir) CC axons were detected at all stages examined, from newborn to adult; the peak density occurred during postnatal weeks (PNW) 3–4. During PNW 1–2, the denisty of NPY-ir CC axons increased gradually; some NPY-ir axons at this age had growth cones located within the CC bundle between the cerebral hemispheres. The density of the NPY-ir CC axons decreased gradually during PNW 5–7, and from PNW 8 to maturity only a few NPY-ir CC axons were observed. These results indicate that at least two types of NPY-ir CC axons (i.e., transitory and permanent) exist during development, and that most of these axons are eliminated or only express NPY-ir for a short period during development. The results also indicate that neurochemical subsets of CC axons participate in the extensive transitory growth observed by means of the membrane tracer DiI but they may follow unique developmental timetables.
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  • 52
    ISSN: 1432-0533
    Keywords: Mitochondrial myopathy ; Ragged red fibers ; In situ hybridization ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In situ hybridization combined with immunohistochemical techniques has been applied to study patients affected by mitochondrial myopathies with large mitochondrial (mt)DNA deletions. All patients' muscle biopsies showed ragged red fibers (RRFs) and cytochrome oxidase (COX) deficiency. Two digoxygenin-labeled, polymerase chain reaction (PCR)-amplifed DNAs were used as probes. One probe was designed to hybridize only with wild-type mtDNAs, while the other recognized both wild-type and deleted mtDNAs. Concomitant immunocytochemical analysis using antibodies against subunits II, III, (encoded by mtDNA) and IV (encoded by nuclear DNA) of COX was carried out. In our patients deleted mtDNAs are overexpressed in COX-negative RRFs, while wild-type mtDNAs are decreased in the same fibers. Immunohistochemistry studies show that COX IV is overexpressed in RRFs and that COX II and COX III subunits are still present. Deleted mtDNAs are spatially segregated in muscle fibers, where they interfere with the local population of normal mitochondrial genomes, causing a regional deficiency of the mitochondrial respiratory activity.
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  • 53
    ISSN: 1432-0533
    Keywords: Key words Amyloid angiopathy ; Enzymehistochemistry ; Hereditary cerebral hemorrhage with amyloidosis (Dutch) ; Immunohistochemistry ; Senile plaques
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Plaque-like lesions and amyloid angiopathy were investigated in the frontal cerebral cortex of four patients with hereditary cerebral hemorrhage with amyloidosis (Dutch) (HCHWA-D), using immunohistochemical [antibodies to β amyloid protein (Aβ), β protein precursor (βPP), synaptophysin, ubiquitin (UBQ), cathepsin D, paired helical filaments (PHF) and glial fibrillary acidic protein (GFAP)], enzymehistochemical (acid phosphatase) and silver [methenamine silver (MS) and Palmgren] staining methods. Whereas Aβ- and MS-positive diffuse plaques were found in all patients, only the three older patients showed neuritic or congophilic plaques, which were acid phosphatase and cathepsin D positive and contained βPP-, synaptophysin- and UBQ-positive, but PHF-negative neurites. These plaques were surrounded by reactive astrocytes. Similar immuno- and enzymereactivity was found around congophilic blood vessels. Thus, apart from neuronal degeneration in a subset of plaque-like lesions and around blood vessels, this study shows an age-related morphology of the plaques in HCHWA-D, corresponding to that in Down's syndrome (DS), with the difference that neurofibrillary (NF) pathology is absent in HCHWA-D in contrast to DS. HCHWA-D may be considered as a model for congophilic plaque formation not associated with NF pathology.
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  • 54
    ISSN: 1432-0533
    Keywords: Perineurial cells ; Nerve regeneration ; Immunohistochemistry ; Epithelial membrane antigen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Perineurial cells are specialized connective tissue cells that form a barrier between endoneurium and epineurium in normal nerves. In the present study, the formation of the perineurium after transection of rat sciatic nerves was investigated. The cord bridging the gap between proximal and distal stumps through silicone tubes was studied 3, 7, 12, 18, and 21 days after surgery using electron microscopy and antibodies against epithelial membrane antigen (EMA), a marker for perineurial cells that has thus far not been applied to the study of differentiating cells in nerve tubulation systems. Initially, a thin cord consisting of fibrin bridged the gap between the stumps. At 7 days, longitudinal cells had migrated from both stumps toward the center of the tubes on the surface of the fibrin cord. These cells were immunoreactive with anti-EMA. At 12 days, ultrastructural features of perineurial cells (desmosomes, tight junctions, actin filaments with dense bodies, tonofilaments) were prominent in these cells. Subsequently, the gap was bridged through the perineurial tube by endothelial cells, pericytes, fibroblasts, Schwann cells, and axons. At 21 days, a single large nerve fascicle ensheathed by a mature perineurium was found between the stumps. Thus, the first cells to connect proximal and distal stumps in the investigated nerve regeneration silicon chamber system are perineurial cells. Through the tube formed by these cells, blood vessels and nerve fibers bridge the gap. Therefore, establishment of a perineurial connection between nerve stumps appears to be important in the sequence of events during nerve regeneration.
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  • 55
    ISSN: 1432-0533
    Keywords: Amyloid angiopathy ; Enzymehistochemistry ; Hereditary cerebral hemorrhage with amyloidosis (Dutch) ; Immunohistochemistry ; Senile plaques
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Plaque-like lesions and amyloid angiopathy were investigated in the frontal cerebral cortex of four patients with hereditary cerebral hemorrhage with amyloidosis (Dutch) (HCHWA-D), using immunohistochemical [antibodies to β amyloid protein (Aβ), β protein precursor (βPP), synaptophysin, ubiquitin (UBQ), cathepsin D, paired helical filaments (PHF) and glial fibrillary acidic protein (GFAP)], enzymehistochemical (acid phosphatase) and silver [methenamine silver (MS) and Palmgren] staining methods. Whereas Aβ-and MS-positive diffuse plaques were found in all patients, only the three older patients showed neuritic or congophilic plaques, which were acid phosphatase and cathepsin D positive and contained βPP-, synaptophysin-and UBQ-positive, but PHF-negative neurites. These plaques were surrounded by reactive astrocytes. Similar immuno-and enzymereactivity was found around congophilic blood vessels. Thus, apart from neuronal degeneration in a subset of plaque-like lesions and around blood vessels, this study shows an age-related morphology of the plaques in HCHWA-D, corresponding to that in Down's syndrome (DS), with the difference that neurofibrillary (NF) pathology is absent in HCHWA-D in contrast to DS. HCHWA-D may be considered as a model for congophilic plaque formation not associated with NF pathology.
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  • 56
    ISSN: 1432-0568
    Keywords: Cerebellum ; Neurological mutant ; Ganglioside ; Immunohistochemistry ; Neuron migration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The cerebellum is organized into a series of parasagittally aligned bands that may be revealed histologically in the adult mouse by largely complementary immunostaining of Purkinje cell sets with the monoclonal antibodies Zebrin II (ZII; antigen:aldolase C) and P-path (PP; antigen:90-acetyl glycolipids). We compared the normal staining pattern using these markers and an antibody to calbindin with that found in the reeler mutants (rl/rl), in which most Purkinje cell migration is halted beneath the cerebellar white matter. The results revealed that Purkinje cells in reeler mutants, despite their ectopic location in large subcortical masses, show a clear tendency to distribute into alternating zones that either stain for Zebrin II or for P-path, with variable transition zones of mixed labeling. However, the estimated number of zones was fewer than in the normal adult cortex: roughly 7–9 zones are revealed per side in the mutant compared with 14 major divisions in wild type mice. These results raise the possibility that neurons destined to express these markers are segregated during their migration and that the final phase of migration into the cortex might involve further splitting or interdigitation between cell sets expressing the two antigens.
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  • 57
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 190 (1994), S. 251-261 
    ISSN: 1432-0568
    Keywords: C-CAM ; Immunohistochemistry ; In situ hybridization ; Palate formation ; Retinoic acid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract C-CAM is a cell surface glycoprotein that is involved in cell adhesion and may play a role in histogenesis and organogenesis. It is a member of the carcinoembryonic antigen (CEA) gene family, which is a subfamily of the immunoglobulin gene superfamily. We have analyzed the expression of C-CAM during normal and disturbed craniofacial development in the mouse by immunohistochemistry and in situ hybridization. Developmental disturbances were induced by retinoic acid (RA) treatment of pregnant mice. Normal and malformed fetuses were examined on days 14, 15, 16, 17 and 18 of gestation. The expression of C-CAM was detected first at day 16. With age, the signal became gradually stronger. C-CAM was detected in the epithelia of both ectodermal and mesodermal origin, including oral and respiratory epithelia, epithelia of the developing vessels, glands and their ducts. In the RA-treated fetuses, the expression of C-CAM was higher in the epithelium of the oral cavity than in that of the nasal cavity, with a distinct borderline between differentiating nasal and oral epithelium of the palatal shelves. However, the submucosal nasal glands and ducts showed higher expression than oral glands in both normal and RA-treated mice. The expression of C-CAM did not differ significantly between control and RA-treated animals. The presence of C-CAM in all proliferating craniofacial epithelia indicates that this molecule may play an important role in development.
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  • 58
    Electronic Resource
    Electronic Resource
    Springer
    Archives of gynecology and obstetrics 255 (1994), S. 181-187 
    ISSN: 1432-0711
    Keywords: Endometriosis ; Estrogen- and progesterone receptors ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract 30 formalin-fixed and paraffin embedded and 20 fresh frozen samples of endometriotic tissue were analysed immunohistochemically for the concentration of estrogen and progesterone receptors. In the formalin-fixed and paraffin embedded group only 37% of the samples were estrogen receptor positive whereas 63% were receptor negative. In contrast, we found that 67% of the samples had a positive progesterone receptor status. In the fresh frozen group 60% of endometriotic tissues were estrogen receptor positive and 75% of the tissues had a positive progesterone receptor status. We could not find any correlation between the site or severity of the endometriosis or the hormonal receptor status. We were able to demonstrate that the immunohistochemical detection of hormonal receptors in endometriotic tissues is possible and that better results were obtained if fresh frozen rather than formalin-fixed and paraffin embedded tissues were analyzed.
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  • 59
    ISSN: 1432-0738
    Keywords: Key words Cell proliferation ; Pancreas ; 5-Bromo-2′-deoxyuridine (BrdU) ; Immunohistochemistry ; Hormones ; Insulin ; Glucagon ; Somatostatin ; Phenobarbitone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  A previous study demonstrated that administration of phenobarbitone to male AP Wistar rats for up to 7 days caused alterations in labelling indices (LIs) in several different tissues (including a reduction of the endocrine pancreas population LI) as determined by immunohistochemical visualisation of 5-bromo-2′-deoxyuridine (BrdU) incorporation into S-phase nuclei. The primary objective of this study was to determine whether treatment with phenobarbitone influenced the replicative states of specific cohorts of the islet (of Langerhans) cell population or generated a uniform depression of LI. Quantitation of the LIs of individual islet cell cohorts was achieved by utilisation of a dual immunohistochemical staining method for BrdU and islet hormones (insulin, glucagon and somatostatin) using a sequential peroxidase anti-peroxidase (PAP)/alkaline phosphatase anti-alkaline phosphatase (APAAP) method employing diaminobenzidine and New Fuchsin chromogens, respectively. We observed reductions, increases and no change in LIs of insulin-, glucagon- and somatostatin-positive cells, respectively. We conclude that the decreased LI of the insulin-positive cohort was not countered entirely by the LI increase in the glucagon-positive cohort due to the larger size of the former. Furthermore, the effects of phenobarbitone treatment are not manifested generally in the islet cell population but in the insulin- and glucagon-positive cohorts only. The causation of these effects is unknown but is likely to be due to enhanced carbohydrate and hormone metabolism. We believe that the visualisation and quantitation of replicating cells in specific hormone-positive cohorts of the islet cell population provide opportunities for understanding the influence of xenobiotics and disease processes on pancreatic function.
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  • 60
    ISSN: 1432-1106
    Keywords: Immunohistochemistry ; Brain proteins ; ChAT ; GFAP ; Memory ; Astrocytes ; “Cholinergicrich” transplants ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In adult, lesion-impaired rat brain receiving embryonic day 15 (E15) fetal transplants, the level of expression of glial fibrillary acidic protein (GFAP) correlates positively with choline acetyltransferase (ChAT) levels and also with measurements of successful behavioural recovery. These results suggest that glial cells may play a pivotal role in the cognitive success of socalled cholinergic-rich transplants.The objective of this study was to investigate the association between GFAP-and ChAT-staining antigens in or around cholinergicrich fetal grafts transplanted in adult cortex. An immunohistochemical fluorescent double-labelling technique was used to simultaneously identify GFAP- and ChAT-staining cells to assess whether there was a different type or distribution of cells present in these successful transplants. On brain sections of transplant area, GFAP-staining glial cells did not co-label with ChAT-staining cells. The transplant area, therefore, did not reveal a different type of cell from those seen in comparable normal cortical brain but rather a greater concentration of both GFAP- and ChAT-positive staining cells.
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  • 61
    ISSN: 1432-0711
    Keywords: Filaggrin ; Immunohistochemistry ; Squamous differentiation ; Uterine Cervix
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Epithelial distribution of filaggrin, a histidine-rich protein related to squamous terminal differentiation, was investigated in 87 cervical biopsies using an avidin-biotin-peroxidase technique with a monoclonal anti-human filaggrin antibody (AKH1). Normal squamous cervical epithelium exhibited a positive homogeneous immunoperoxidase stain in the upper parabasal, intermediate and superficial cell layers. Similar findings were obtained in cervical condylomas, although full-thickness staining was observed in 35.7% of the cases (P〈0.001). Filaggrin expression in CIN was inversely related to the severity of the lesion (P〈0.001). An irregular staining pattern was present in most high-grade CIN. Filaggrin expression was closely connected to the degree of tumour differentiation (P〈0.05) in squamous cell carcinomas of the cervix. Abnormal filaggrin stainings identified a premalignant/malignant cervical squamous lesion with a positive predictive value of 92.3%. Non-squamous epithelia showed lack of filaggrin expression. Filaggrin may therefore be considered a marker of squamous differentiation in both the normal and pathological human uterine cervix.
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  • 62
    ISSN: 1432-069X
    Keywords: Psoriasis ; Adhesion receptors ; CLA ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Adhesion receptors and their ligands play a vital role in the immune system. We studied the expression of different adhesion receptors, using single- and double-staining immunohistochemical techniques, in both lesional and non-lesional skin specimens from seven psoriasis patients and in skin biopsy specimens from eight normal healthy controls. Our results showed an overall increased expression of several adhesion receptors in both lesional and non-lesional psoriatic skin. We consistently found an increased expression in particular of ICAM-1 and E-selectin on endothelial cells, and ICAM-1 on T cells and Langerhans cells. In contrast, a weak expression of VCAM-1 was found on endothelial cells and mononuclear cells in lesional psoriatic skin specimens alone. Interestingly, LFA-1 was also expressed on Langerhans cells, with a greater frequency in skin from lesional than from non-lesional sites, but was never expressed in skin from normal healthy individuals. Furthermore, significantly increased numbers of Langerhans cells and T cells with a positive reactivity for MAb HECA-452 were found in both lesional and non-lesional psoriatic skin. We hypothesize that the enhanced expression of adhesion receptors on migrating immunocompetent cells and endothelial cells of psoriatic skin in general facilitates the increased influx of activated T lymphocytes and other immunocomponent cells into the skin, and thus underscores the generalized character of the disease.
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    Amino acids 6 (1994), S. 1-13 
    ISSN: 1438-2199
    Keywords: Amino acids ; β-Thymosins ; Phylogenetic distribution ; Actin sequestration ; Immunoassays ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary β-Thymosins, a group of highly homologous peptides consisting of about 40 amino acid residues, were found to be distributed from mammals up to echinoderms. Althogh they have first been isolated from mammalian thymus tissue preparations, their occurrance is not organ-specific and they are present even in different types of cells. For thymosinβ 4 several biological activities have been reported, stating that this peptide acts as a thymus peptide hormone and is also involved in the neuroendocrine and immune system. However, it was recently demonstrated that thymosinβ 4 has actin-sequestering properties and therefore might play an important role in the regulation of the microfilament system. This fact gives a new outlook on the real biological function ofβ-thymosins.
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    International journal of legal medicine 107 (1994), S. 132-140 
    ISSN: 1437-1596
    Keywords: Time of death ; Sweat glands ; Immunohistochemistry ; Electron microscopy ; Todeszeit ; Schweißdrüsen Immunhistochemie ; Elektronenmikroskopie
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Law
    Description / Table of Contents: Zusammenfassung Diese Untersuchung zeigt postmortale autolytische Veränderungen in der Haut auf zellulärer und subzellulärer Ebene und identifiziert Parameter, welche helfen können, die Zeit des Todes in den ersten Stunden postmortem zu bestimmen. Hautproben von der Beugeseite des Arms wurden, 3, 6, 9 und 12. Stunden nach dem Tode von insgesamt 29 Leichen entnommen (verschiedene Altersklassen, keine Zeichen für Hauterkrankungen, verschiedene Todesursachen). Drei Arten der Untersuchungen wurden durchgeführt: zytochemisch (Hematoxylin-Eosin and Alcian-PAS), immunhistochemisch (S-100, CEA, Cytokeratin, ASM) und ultrastrukturell (Elektronenmikroskopie). Die Elektronenmikroskopie erwies sich als nützlich für die Identifizierung von Transformationen die für jeden chronologischen Schritt spezifisch waren: Reduktion des intrazellulären Glykogens in hellen Zellen und Reduktion der sekretorischen Granula in dunklen Zellen sind typische Zeichen für die erste Phase (3 Stunden) nach dem Tode; mitochondriale Dilatation und Rarifizierung der Cristae in hellen und dunklen Zellen sind typisch für die 2. Phase (6 Stunden); Rarifizierung der Microvilli in dunklen und hellen Zellen sind typisch für die 3. Phase (9 Stunden) und Kernpyknose von dunklen und hellen Zellen ist ein Zeichen der letzten Phase (12 Stunden). Zytochemie und Immunhistochemie sorgen für eine nützliche Information — dies gilt nicht für alle chronologischen Stadien, welche hier einbezogen wurden, aber für individuelle Phasen (3 Stunden für Hematoxylin-Eosin und 6 Stunden für Alcian-PAS). Es ist jedoch besonders wichtig, die Resultate von allen solchen Techniken simultan einzubeziehen, so daß die Frage der exakten Todeszeit innerhalb der ersten 12 Stunden postmortem genauer beantwortet werden kann.
    Notes: Abstract This study demonstrates post-mortem autolytic alterations in the skin at cellular and subcellular levels and identifies parameters which may assist in determining the time of death in the first few hours post-mortem. Serial skin samples from the ventral surface of the arm were taken at intervals of 3, 6, 9 and 12 h after death in 29 subjects of various ages, with no signs of skin disease; causes of death were various. Three types of tests were performed: cytochemical (hematoxylin-eosin and alcian-PAS), immunohistochemical (S-100, CEA, Cytokeratin, ASM) and ultrastructural (electron microscopy). Electron microscopy proved useful for identifying transformations which were found to be specific for each chronological step considered: reduction of intracellular glycogen in clear cells and reduction of secretory granules in dark cells are typcial signs of the first stage (3 h) after death; mitochondrial dilatation and rarefaction of cristae in clear and dark cells are typical of the second stage (6 h); rarefaction of microvilli in dark and clear cells is a sign of the last stage (12 h). Cytochemistry and immunohistochemistry supply useful information — not for all the chronological stage considered here, but for individual phases (3 h for hematoxylin-eosin and 6 h for alcian-PAS). However, it is particularly important to use the results from all such techniques simultaneously, so that the question of the exact time of death within the first 12 h post-mortem may be more accurately answered.
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    Rheumatology international 13 (1994), S. 181-186 
    ISSN: 1437-160X
    Keywords: Rheumatoid arthritis ; Immunohistochemistry ; Enzyme histochemistry ; Histopathology ; Chronic synovitis ; Macrophages
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Rheumatoid arthritis (RA) is an inflammatory disease of the synovial membrane, which results in the destruction of joints by inflammatory pannus. The synovial membrane shows proliferation and cellular infiltrates on microscopy with signs of chronic and acute inflammation. Macrophages are thought to play a central role in the pathogenesis of RA. We examined synovial membrane specimens of 21 RA patients using morphological, immunohistological and enzyme histochemical methods for number and distribution of macrophages. We were able to identify 41.5±8.8% of lining cells as macrophages, depending on the method used. In abundant diffuse lymphocellular infiltrates, 23.4±11.1% of mononuclear cells were macrophages. In addition, most cells in the region of tumorlike proliferation and a stromal population of fibroblastlike cells were detected by macrophage markers. Although cell number in synovial membrane increases drastically, we did not find correlations between the relative amount of macrophages in these regions and basic activity. Basic activity includes proliferative reaction as well as lymphoplasmacellular and mononuclear infiltration-both signs of an immunopathological process. In contrast, using enzymes or activation markers, there was a clear correlation. We consider that a constant high percentage of macrophages in RA synovial membrane is present regardless of any actual in flammatory process.
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    Journal of cancer research and clinical oncology 120 (1994), S. 672-677 
    ISSN: 1432-1335
    Keywords: Ovary ; Undifferentiated carcinoma ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Immunohistochemical characteristics of undifferentiated carcinomas of the ovary were examined using formalin-fixed, paraffin-embedded tissues with an avidinbiotin staining approach. Eight cases were collected from the pathology files of our Institute from a total of 214 recorded malignant ovarian tumors. For immunostaining, antibodies reacting with epithelial membrane antigen (EMA), pankeratin, vimentin, CA 125, CA 19-9, carcinoembryonic antigen (CEA), α-fetoprotein (AFP), α-l-antitrypsin (AT), epidermal growth factor receptor (EGFR), c-erbB-2,bcl-2 and p53 proteins were used. All the cases examined were positive for EMA and pankeratin, specific markers for epithelial tumors, negative for the non-epithelial tumor marker, vimentin, and also positive for EGFR. Interestingly, only one case was positive for CA 125, despite it being one of the commonest reported indicators of ovarian cancer. CA 19-9 was positive in 7 cases, CEA in 5, AFP in 2, AT in 6 and c-erbB-2 protein in 4. Two cases were positive for p53 protein, and in 1 of these positive staining forbcl-2 was also observed. These results indicate that the epithelial nature is well preserved in undifferentiated ovarian carcinomas, although consistently positive reactions were not observed within the cases for some antigens. They further celarly show that a negative signal for CA 125 can not be considered to exclude the possibility of a primary ovarian tumor.
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    Knee surgery, sports traumatology, arthroscopy 2 (1994), S. 118-122 
    ISSN: 1433-7347
    Keywords: Anterior cruciate ligament ; Allograft transplantation ; Nerve supply ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Sports Science
    Notes: Abstract We investigated the nerve supply of anterior cruciate ligaments ((ACLs) and of cryopreserved bone-ACL-bone allografts in a rabbit model with immunohistochemical methods to establish the distribution pattern of the nervous tissues and to determine the reinnervation rate of ACL allografts. The ACL is innervated by three different classes of nerve fibre: (1) fibres of large diameter, characterized by neurofilament immunoreactivity, which are fast-conducting mechanoreceptive sensory afferents; (2) fibres of small diameter, characterized by substance Pimmunoreactivity, which are slow-conducting nociceptive sensory afferents; and (3) sympathetic efferent vasomotor fibres, characterized by their immunoreactivity to the ratelimiting enzyme of noradrenaline synthesis, tyrosine hydroxylase. The ACLs showed numerous fibres of all three nerve classes; as specialised sensory nerve endings only Ruffini corpuscles were observed. All nerve fibres were located subsynovially, none within the collagen core of the ligament itself. No nerve fibres were detected in the ACL allografts at 3 and 6 weeks. Sparse fibres were detected at 12 weeks, while the 24-, 36-and 52-week specimens showed plenty of all three fibre types. No mechanoreceptors were found in the ACL allografts. To our knowledge, this method for the first time allows a differentiation of the nerve fibres of ACLs and ACL allografts into three different nerve fibre classes with known neurophysiological functions.
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    Documenta ophthalmologica 88 (1994), S. 105-112 
    ISSN: 1573-2622
    Keywords: Immunohistochemistry ; Pathogenesis ; Proliferative vitreous-retinopathy ; Retinal detachment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Proliferative vitreo-retinopathy (PVR) and subretinal membrane proliferation are the most common complication and cause of failure in retinal-detachment (RD) surgery. In this study, material withdrawn from 21 patients was observed. The vitreal taps of 16 bulbs affected by PVR and which had undergone vitrectomy, along with 5 bulbs obtained by enucleation, were stained with Hematoxylin Eosin and studied immunohistochemically. The cells involved in this proliferative tissue include macrophages, cellular elements of pigmented epithelium origin, fibroblasts and myofibroblasts. From the examination of enucleated bulbs, we can easily recognize that the cellular components of the membrane are represented by fibroblasts, capillaries, and occasional macrophages; meanwhile, PE cells remain at the base of the newly formed tissue.
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  • 69
    ISSN: 1434-0879
    Keywords: Urological diseases ; Epidermal growth factor ; Epidermal growth factor receptor ; Transitional cell carcinoma ; Nephrectomy ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract To examine the excretion of urinary epidermal growth factor (EGF) in urological diseases and the relationship of EGF urine levels with transitional cell carcinoma (TCC), we measured the concentration of EGF by radioimmunoassay. The series comprised patients with active TCC (n=50), others in tumor-free status (n=29) and with non-neoplastic inflammatory diseases (n=43), and normal controls (n=50). Urinary EGF values were lower in patients with urological diseases of different etiologies than in normal controls (P〈0.005). Mean EGF levels of patients who had previous bladder tumor resection (n=21) were not statistically different from normal controls (P=0.2). For patients with active TCC, EGF urine levels showed a significant inverse relationship to increasing tumor grade (P=0.02). In addition, subjects who had received nephrectomy for pelvic carcinoma (n=8) showed significantly lower mean EGF values than those with intact kidneys (n=21), irrespective of sex (P〈0.05). Immunostaining of EGF on non-neoplastic kidney (n=9) revealed reactivity in the distal convoluted tubules and thick ascending limbs of Henle. Our results suggest that the kidney is the major source of urinary EGF. Its excretion in urine is decreased in both inflammatory and neoplastic diseases of the urinary tract. EGF may play an important part in the biological activity of TCC. Further study is indicated to investigate the monitoring of EGF urine levels as a marker of recurrence for EGF receptor-positive TCC.
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    European archives of oto-rhino-laryngology and head & neck 251 (1994), S. 137-142 
    ISSN: 1434-4726
    Keywords: Otitis media with effusion ; Ventilation tubes Middle ear collagen ; Immunohistochemistry ; Rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Morphometric quantitation of the area fractions of collagen types I, II, IV and V was determined in the normal rabbit middle ear mucosa and in relation to otitis media with effusion (OME) using a three-layered peroxidase-antiperoxidase technique. The effects of substituting normal low-oxygen middle ear gas (non-ventilated) with atmospheric air (ventilated) were studied in both healthy ears and ears with OME. Based upon previous histological examinations in rabbits, only ears with OME for more than 8 weeks were included to ensure the presence of chronic inflammation (COME). Atmospheric air was introduced into the middle ears by insertion of ventilation tubes or by an enlarged myringotomy. Collagen type I was predominant in all groups studied. The area fractions of collagen types I, II and IV were increased significantly in COME, with collagen type II elevated in particular. Ventilation of the normal ears resulted in a significantly increased area fraction of cells, while the area fractions and distributions of the collagen types were unaffected. None of the ventilated ears in COME improved or healed spontaneously. The total fraction of collagen in COME was not changed significantly by the introduction of atmospheric air. However, the individual distribution of the collagen types was altered, with significantly larger area fractions of types II and V found in ventilated ears with COME. Possible explanations for the differences found are discussed, including the role of oxygen-derived free radicals.
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    Cell Motility and the Cytoskeleton 27 (1994), S. 133-149 
    ISSN: 0886-1544
    Keywords: MAP4 depletion ; antibody blocking ; detyrosination ; midbody ; asymmetric cell processes ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Previous immunolocalization studies using many primate cultured cell lines demonstrated that a microtubule-associated protein of Mr ∼210,000 which is now called MAP4, is present along the length of microtubules in interphase and mitotic cells [Bulinski and Borisy (1980) J. Cell Biol. 87:802-808; DeBrabander et al. (1981) J. Cell Biol. 91:438-455]. Since MAP4 has been implicated as a microtubule stabilizer, we asked whether all classes of microtubules possess an equal complement of MAP4. We have reexamined the cellular distribution of MAP4, using both conventional double-label immunofluorescence and an antibody blocking technique [Schulze and Kirschner (1987) J. Cell Biol. 104:277-288] to highlight microtubules lacking, or depleted in, MAP4. These techniques have revealed that thin processes extending from monkey kidney cells (TC-7), and those made by human neuroblastoma cells (IMR-32) in response to retinoic acid, are often deficient in MAP4 immunoreactivity. Since both types of cellular processes contain stable microtubules, which are enriched in detyrosinated (Glu) tubulin, we tested the ability of MAP4 to bind to microtubules made from pure Glu and pure tyrosinated (Tyr) tubulin in vitro. MAP4 bound to both types of microtubules, and the similar saturation level of MAP4 binding to Glu and Tyr microtubules suggested that differential binding to these forms of tubulin does not contribute directly to a mechanism for segregation of MAP4 on microtubules in vivo. In TC-7 cells, we also observed MAP4-depletion on single microtubules, distal regions of broad cytoplasmic extensions, and midbodies of dividing cells. MAP4 depletion may reflect recent, rapid growth of microtubules to which MAP4 has not yet bound, or the presence of other MAPs that may compete with MAP4 for binding sites on the MT. We suggest that different levels of MAP4 on microtubules may directly modulate microtubule dynamics within single cells, as well as other microtubule functions such as those involving microtubule motor activity. © 1994 Wiley-Liss, Inc.
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    Cell Motility and the Cytoskeleton 27 (1994), S. 193-205 
    ISSN: 0886-1544
    Keywords: amoeboid motility ; fluorescence ratio imaging ; BCECF ; nematodes ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The development and locomotion of the amoeboid sperm of the nematode, Ascaris suum, depend on precise control of the assembly of their unique major sperm protein (MSP) filament system. We used fluorescence ratio imaging of cells loaded with BCECF to show that intracellular pH (pHi) is involved in controlling MSP polymerization in vivo. Spermatogenesis is marked by a cycle of MSP assembly-disassembly-reassembly that coincides with changes in pHi. In spermatocytes, which contain MSP in paracrystalline fibrous bodies, pHi was 6.8, 0.6 units higher than in spermatids, which disassemble the fibrous bodies and contain no assemblies of MSP filaments. Activation of spermatids to complete development resulted in rapid increase in pHi to 6.4 and reappearance of filaments. Treatment of spermatocytes with weak acids caused the fibrous bodies to disassemble whereas incubation of spermatids in weak bases induced MSP assembly. The MSP filaments in spermatozoa are organized into fiber complexes that flow continuously rearward from the leading edge of the pseudopod. These cells established a pseudopodial pH gradient with pHi 0.15 units higher at the leading edge, where fiber complexes assemble, than at the base of the pseudopod, where disassembly occurs. Acidification of these cells caused the MSP cytoskeleton to disassemble and abolished the pH gradient. Acid removal resulted in reassembly of the cytoskeleton, re-establishment of the pH gradient, and re-initiation of motility. MSP assembly in sperm undergoing normal development and motility and in cells responding to chemical manipulation of pHi occurs preferentially at membranes. Thus, we propose that filament assembly in sperm is controlled by pH-sensitive MSP-membrane interaction. © 1994 Wiley-Liss, Inc.
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    Cell Motility and the Cytoskeleton 27 (1994), S. 248-261 
    ISSN: 0886-1544
    Keywords: myogenesis ; protein isoforms ; muscle types ; Z-disc ; phosphorylation ; chicken muscle ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The morphogenesis of functional myofibrils in chick skeletal and cardiac muscle occurs in greatly different time spans, in about 7 and 2 days, respectively. In chick skeletal myogenic cells, one isoform of the 250 kD actin-binding protein (ABP) filamin is associated with stress fiber-like structures of myoblasts and early myotubes, then disappears for approximately 4 days, whereupon a second filamin isoform reappears at the Z-disc periphery. We sought to determine if cardiac myogenesis involves this sequence of appearance, disappearance, and reappearance of a new filamin isoform in a compressed time scale. It was known that in mature heart, filamin is localized at the Z-disc periphery as in mature (fast) skeletal muscle, and is also associated with intercalated discs. We find that myocardial filamin has an apparent molecular weight similar to that of adult skeletal muscle filamin and lower than that of smooth muscle filamin, and that both skeletal and cardiac muscle contain roughly 200 filamin monomers per sarcomere. Two-dimensional peptide mapping shows that myocardial filamin is very similar to skeletal muscle filamin. Myocardial, slow skeletal, and fast skeletal muscle filamins are all phosphorylated, as previously shown for filamin of non-striated muscle. Using immunofluorescence, we found that filamin could not be detected in the developing heart until the 14-somite stage, when functional myofibrils exist and the heart has been beating for 3 to 4 hours. We conclude that in cardiac and skeletal myogenesis, different sequences of filamin gene expression result in myofibrils with similar filamin distributions and isoforms. © 1994 Wiley-Liss, Inc.
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    Cell Motility and the Cytoskeleton 27 (1994), S. 272-283 
    ISSN: 0886-1544
    Keywords: cell cycle ; transcription ; mRNA decay ; autoregulation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The single alpha-tubulin gene of Tetrahymena thermophila was isolated from a genomic library and shown to encode a single protein. Comparisons of the rates of evolution of this gene with other alpha-tubulin sequences revealed that it belongs to a group of more evolutionarily constrained alpha-tubulin proteins in animals, plants, and protozoans versus the group of more rapidly evolving fungal and variant animal alpha-tubulins. The single alpha-tubulin of Tetrahymena must be used in a variety of microtubule structures, and we suggest that equivalently conserved alpha-tubulins in other organisms are evolutionarily constrained because they, too, are multifunctional. Reduced constraints on fungal tubulins are consistent with their simpler microtubule systems. The animal variant alpha-tubulins may also have diverged because of fewer functional requirements or they could be examples of specialized tubulins. To analyze the role of tubulin gene expression in regulation of the complex microtubule system of Tetrahymena, alpha-tubulin mRNA amounts were examined in a number of cell states. Message levels increased in growing versus starved cells and also during early stages of conjugation. These changes were correlated with increases in transcription rates. Additionally, alpha-tubulin mRNA levels oscillate in a cell cycle dependent fashion caused by changes in both transcription and decay rates. Therefore, as in other organisms, Tetrahymena adjusts alpha-tubulin message amounts via message decay. However the complex control of alpha-tubulin mRNA during the Tetrahymena life cycle involves regulation of both decay and transcription rates. © 1994 Wiley-Liss, Inc.
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    Cell Motility and the Cytoskeleton 28 (1994), S. 45-58 
    ISSN: 0886-1544
    Keywords: MTOC ; cytoplasmic microtubule complex ; antitubulin ; Immunofluorescence ; ultrastructure ; immunogold ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We investigated the microtubule (MT) cytoskeleton and microtubule centers (MTC) in undifferentiated amoebae by indirect immunofluorescence with six monoclonal antitubulin antibodies, and by transmission electron microscopy and immunogold ultracytochemistry. Interphase amoebae of both species contain a distinct cytoplasmic complex of MTs, which is more elaborate in Protostelium mycophaga. In Acytostelium leptosomum amoebae a single MTC is attached to each interphase nucleus at its pointed end, as in the other dictyostelid cellular slime molds Dictyostelium discoideum and Polysphondylium violaceum. Ultrastructurally, MTCs of A. leptosomum also resemble those of these two species: They consist of an electron-opaque core shaped like a stout rod, which is embedded, together with nodules, in a fuzzy matrix. The nodules are the points of origin of the MTs. In most amoebae of P. mycophaga there are two MTCs on opposite sides of and close to the nucleus, but many amoebae also contain a variable number of MTCs that are remote from the nucleus. Nucleus-associated and “remote” MTCs are structurally identical. They consist of a ring-shaped core with inner and outer diameters of ca. 130 nm and 340 nm. A plug sits in the ring, and satellites are connected to the core by fine fibrils. The satellites are the points of origin of MTs. New MTCs are apparently formed during mitosis, the parent MTC probably serving as a template for the genesis of a new ring. The results support the notion that phylogenetically related organisms have similarly constructed MTCs and that these are dissimilar in less closely related organisms. © 1994 Wiley-Liss, Inc.
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    Cell Motility and the Cytoskeleton 28 (1994), S. 135-142 
    ISSN: 0886-1544
    Keywords: bidirectional swimming ; flagellar movement ; helical bends ; 9+0 axoneme ; planar bends ; viscosity ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Spermatozoa of the small myzostomid worm Myzostomum cirriferum usually swim with the flagellum foremost but occasionally stop and then swim with the head foremost. The spermatozoa have axoneme of the 9+0 type; thus each lacks the central pair microtubules. The flagellum emerges in the anterior end of the cell body and attaches to it with junctions. To understand the mechanism regulating the swimming direction of the spermatozoa, we recorded the sperm and their flagellar movements using a video camera with a high-speed shutter. The effects of calcium and viscosity on these movements were also examined.The cell body with the flagellum attached to it formed a curved plate during beating, while the free portion of the flagellum beats with small helical bends. Motive force to propel a spermatozoon was mainly due to the bends in the cell body. The spermatozoa reversed the direction of their swimming as a result of a change in the direction of bend propagation. The direction of bend propagation was regulated by calcium; the bends in the cell body propagated from the end of the head toward the free portion of the flagellum at low concentrations of Ca2+, whereas the direction of bend propagation was reversed at high concentrations of this ion. High viscosity of the medium stimulated a change in the direction of bend propagation. © 1994 Wiley-Liss, Inc.
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    Cell Motility and the Cytoskeleton 28 (1994), S. 155-164 
    ISSN: 0886-1544
    Keywords: microfilamentous cytoskeleton ; actin binding proteins ; formyl peptides ; ionic extraction ; immunoblots ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: F-actin is a major component of the neutrophil (PMN) cytoskeleton. In basal PMNs, F-actin exists in two structurally and functionally distinct pools: Triton insoluble F-actin (TIF)-cold insensitive, not depolymerizable by dilution, and distributed in pseudopods and submembranous locations; and Triton soluble F-actin (TSF)-unstable in cold, diffusely distributed, and gelsolin enriched. The element(s) conferring these unique properties to the Triton insoluble F-actin pool are unknown, but logically include distinct actin regulatory proteins. To study the morphologic and functional determinants of the Triton insoluble F-actin pool, the distribution and quantity of three candidate regulatory proteins, α-actinin, tropomyosin (TM), and actin binding protein (ABP-280), were compared in F-actin (Triton insoluble and Triton soluble) and G-actin pools isolated from basal and chemotactic factor activated human PMNs in suspension, using immunoblots and ionic extraction. F-actin content was measured by NBDphallacidin binding and gel scans. The results show that: (1) α-actinin, actin binding protein 280, and tropomyosin are localized to TIF and excluded from TSF; (2) TM, α-actinin, and ABP 280 are required to stabilize fractions of Triton insoluble F-actin in PMNs; and (3) chemotactic factor activation results in release of a fraction of TM from the Triton insoluble F-actin pool in temporal association with F-actin polymerization in the Triton insoluble F-actin pool. Shifts in ABP 280 or α-actinin do not occur. The results suggest that TM, α-actinin, and ABP 280 provide structure to TIF and that TM release from TIF is involved in chemotactic factor induced actin polymerization in PMNs. © 1994 Wiley-Liss, Inc.
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    Cell Motility and the Cytoskeleton 28 (1994), S. 165-178 
    ISSN: 0886-1544
    Keywords: WISH ; Keratin ; 3-D reconstruction ; mitosis ; intermediate filaments ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Three dimensional (3-D) reconstruction of four mitotic WISH cells from ultrathin sections gave an informative representation of the spatial distribution of keratin densities in these cells. The correspondence between the densities as studied by transmission electron microscopy (TEM) and the Keratin bodies initially revealed by immunoflourescent colabeling of cultures, was confirmed by immunoelectron-microscopy. The smaller, and sometimes more elongated densities, were relatively abundant just beneath the subplasmalemmal microfilament band; and at certain levels of the mitotic cell they were observed to be connected to neighboring densities by intact intermediate filaments (IFs). The larger and more spherical densities appeared to be somewhat more discrete and randomly distributed. Other observed associations of the keratin densities included the telophase contractile ring of microfilaments, chromosomes, the reformed telophase nucleus, and desmosomal junctions with neighboring interphase cells. Cytochalasin D (CD) treatment of cells displaced the peripheral keratin densities toward the cell membrane. The density volume constituted 0.52% to 1.57% of the total cell volume, and the proportional density size was decreased in the cells that had progressed into anaphase and telophase. The observed formation and subsequent dissolution of keratin densities during mitosis may represent a dynamic mechanism of restructuring the keratin cytoskeleton in an unpolymerized form in order to allow for rapid reformation of interphase cell junctions. The physical associations observed between intact IFs and the keratin densities may provide support at certain depths of the mitotic cell, and the juxtaposition of densities with nuclear components suggests a possible source of and role for keratin IFs during nuclear events. © 1994 Wiley-Liss, Inc.
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  • 79
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    Cell Motility and the Cytoskeleton 28 (1994), S. 199-204 
    ISSN: 0886-1544
    Keywords: axoneme ; cilia ; flagella ; microtubule ; motility ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Observations that were interpreted to provide evidence for equivalent functions of all axonemal dyneins should be reinterpreted, and models based on this assumption should be abandoned. In the future, attempts to understand the mechanisms for flagellar bending, oscillation, and bend propagation should start from the assumption that each type of axonemal dynein may have a specific function. At least three distinct functions can now be identified: bend initiation, maintenance of the angle of propagating bends, and generation of power to overcome viscous resistances. Only the last of these three functions is an outer arm dynein function. © 1994 Wiley-Liss, Inc.
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  • 80
    ISSN: 0886-1544
    Keywords: cytoskeleton ; actin binding ; transgelin sequence ; gelation ; gene family ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We have used degenerate oligonucleotides, derived from the amino acid sequence of transgelin peptides [Shapland et al., 1993: J. Cell Biol. 121:1065-1073], to isolate and sequence overlapping cDNA clones encoding this actin gelling protein. Primers with 5′ restriction enzyme sites directed against the N and C terminal amino acids present in these clones were then used to amplify and clone the entire transgelin coding region from reverse transcribed rat small intestine cDNA (RT-PCR). These studies have shown that transgelin is the product of a single gene which is conserved between yeast, Drosophila, molluscs, and humans. Transgelin is expressed as a single message that is regulated at the level of transcription in SV40 transformed 3T3 cells. Our data have shown that transgelin and several other proteins of unknown function, SM22α [Pearlstone et al., 1987: J. Biol. Chem. 262:5985-5991], mouse p27 [Almendral et al., 1989: Exp. Cell Res. 181:518-530], and human WS3-10 [Thweatt et al., 1992: Biochem. Biophys. Res. Commun. 187:1-7], share extensive homology. More limited regions of homology shared between transgelin and other proteins such as rat NP25 (unpublished), chicken calponins α and β [Takahashi and Nadal-Ginard, 1991: J. Biol. Chem. 266:13284-13288], and Drosophila mp20 [Ayme-Southgate et al., 1989: J. Cell Biol. 108:521-531] suggest that all of these proteins may be classified as members of a new transgelin multigene family. © 1994 Wiley-Liss, Inc.
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  • 81
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    Cell Motility and the Cytoskeleton 28 (1994), S. 279-284 
    ISSN: 0886-1544
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 82
    ISSN: 0886-1544
    Keywords: cytoskeleton ; cell culture ; gene expression ; Northern blot ; serum-induction ; rat ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Cytochalasin D and dBcAMP cause cultured astrocytes to change from flat cells to retrated process-bearing cells. F-actin was present throughout cells stimulated with dBcAMP for 16 h, whereas cytochalasin D caused F-actin to form massive aggregates at the tips of the cell processes. The two drugs differently regulated the expression of both β-actin and tropomyosin genes in astrocytes cultured in the presence or absence of serum: dBcAMP caused down-regulation and cytochalasin D caused up-regulation. Northern blot analyses indicated that: (1) serum deprivation halved the concentration of all tropomyosin transcripts (TM-1, TM-2, TM-4, TMBr-1, TMBr-2). Serum induced TM-4 via transcriptional activation, independent of protein synthesis, (2) dBcAMP induced down-regulation of β-actin (-50%) and tropomyosin transcripts (-35 to 52%) even in the presence of serum. The concentration of profilin mRNA decreased in dBcAMP-reactive astrocytes (-46%). The decrease in β-actin mRNA concentration was not blocked by cycloheximide, whereas down-regulation of tropomyosin transcripts was completely reversed when protein synthesis was inhibited, and (3) cytochalasin D induced an increase in the concentration of tropomyosin transcripts (+ 69 to 185%) which was cumulative with serum stimulation. Cytochalasin D induction of both β-actin and TM-4 operated through transcriptional activation, independent of protein synthesis.The production of all tropomyosin transcripts examined here were strictly coordinated with β-actin expression in serum-, dBcAMP- and cytochalasin D-treated astrocytes. This indicates that the differential expression of tropomyosin isoforms occurring during astrocyte maturation is due to more complex regulation than that involved in serum- or cAMP-stimulated astrocytes. © 1994 Wiley-Liss, Inc.
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  • 83
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    Cell Motility and the Cytoskeleton 28 (1994), S. 333-345 
    ISSN: 0886-1544
    Keywords: ciliary beat frequency ; metachronal wave ; ciliary coupling ; extracellular ATP ; acetylcholine ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: In the present work we measured in real time the metachronism and degree of correlation between beating cilia from cultured mucociliary epithelium. The method is based on simultaneous measurement of ciliary beat frequency, phase shifts, and correlation factors in two directions: parallel and perpendicular to the effective stroke direction (ESD). From the phase shifts the lengths of wave components, and consequently the metachronal wavelength and direction, were evaluated.On active ciliary areas of cultured frog esophagus under normal conditions, a relatively high degree of correlation is observed, but cilia are more correlated in direction parallel to ESD which is also the direction of the mucus propulsion. The length of the wave component parallel to ESD is more than twice as large as that of the perpendicular component. The metachronal wavelength was found to be in the range of 5-9 μm, and the direction of the wave propagation was in the range of 90°-125° clockwise to the ESD.When ciliary beat frequency was rapidly increased by extracellular ATP or acetylcholine, only minor effects were observed on the degree of correlation between beating cilia. The length of the wave component parallel to ESD showed the most dramatic effect increasing up to tenfold. The perpendicular to ESD component was not affected by the stimulation. Consequently, the metachronism became more laeoplectic with the angle between the ESD and the wave directions decreasing by 10°-30°, and the metachronal wavelength remained unaltered. © 1994 Wiley-Liss, Inc.
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  • 84
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    Cell Motility and the Cytoskeleton 29 (1994), S. 57-71 
    ISSN: 0886-1544
    Keywords: microtubule bundling ; cytoskeleton ; tau ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Microtubule protein extracted from dogfish erythrocyte cytoskeletons by disassembly of marginal bands at low temperature formed linear microtubule (MT) bundles upon reassembly at 22°C. The bundles, which were readily visible by video-enhanced phase contrast or DIC microscopy, increased in length and thickness with time. At steady state after 1 hour, most bundles were 6-11 μm in length and 2-5 MTs in thickness. No inter-MT cross-bridges were visible by negative staining. The bundles exhibited mechanical stability in flow as well as flexibility, in this respect resembling native marginal bands. As analyzed by SDS-PAGE and immunoblotting, our standard extraction conditions yielded MT protein preparations and bundles containing tau protein but not high molecular weight MAPs such as MAP-2 or syncolin. In addition, late fractions of MT protein obtained by gel filtration were devoid of high molecular weight proteins but still produced MT bundles. The marginal band tau was salt-extractable and heat-stable, bound antibodies to mammalian brain tau, and formed aggregates upon desalting. Antibodies to tau blocked MT assembly, but both assembly and bundling occurred in the presence of antibodies to actin or syncolin. The MTs were “unbundled” by subtilisin or by high salt (0.5-1 M KCl or NaCl), consistent with tau involvement in bundling. High salt extracts retained bundling activity, and salt-induced unbundling was reversible with desalting. However, reversibility was observed only after salt-induced MT disassembly had occurred. Reconstitution experiments showed that addition of marginal band tau to preassembled MTs did not produce bundles, whereas tau presence during MT reassembly did yield bundles. Thus, in this system, tau appears to play a role in both MT assembly and bundling, serving in the latter function as a coassembly factor. © 1994 Wiley-Liss, Inc.
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  • 85
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    Cell Motility and the Cytoskeleton 29 (1994), S. 82-93 
    ISSN: 0886-1544
    Keywords: trichomonads ; cytoskeleton ; antibodies ; electrophoresis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The production of monoclonal antibodies and the use of biochemical techniques revealed that B-type costa proteins in trichomonads are composed of several major polypeptides with molecular weight detected between 100 and 135 kDa similar to those found in the A-type costae. Although differences were observed between the two types in their fine structure, we tested whether proteins composing the two costa types belong to the same protein family. A polyclonal antibody produced against the 118 kDa costa protein of Trichomonas vaginalis also recognized a 118 kDa costa protein in all other trichomonad genera studied so far whether they have A- or B-type costae. Moreover biochemical characteristics of costa proteins indicated that these proteins might represent a novel class of striated root-forming proteins in addition to centrin, giardin, and assemblin. © 1994 Wiley-Liss, Inc.
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  • 86
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    Cell Motility and the Cytoskeleton 29 (1994), S. 110-116 
    ISSN: 0886-1544
    Keywords: high-molecular weight MAPs ; microfilaments ; microtubules ; low-shear viscometry ; taxol ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: High molecular weight microtubule-associated proteins MAP1A and MAP2 form thin projections from microtubule surfaces and have been implicated in crosslinking microtubules and other cytoskeletal components. We have purified native MAP1A from bovine brain and have studied its interaction with G- and F-actin. Using a solid-phase immunoassay we show that MAP1A binds in a dose-dependent manner to both G-actin and F-actin. Addition of MAP1A to F-actin causes gelation of F-actin and SDS-PAGE analysis shows that MAP1A co-sediments with the gelled network, under conditions where F-actin alone does not pellet. The low apparent viscosity of F-actin is markedly increased in the presence of MAP1A, suggesting that MAP1A can crosslink F-actin. Co-incubation experiments indicate that MAP1A and MAP2 may bind to common or overlapping sites on the actin molecule. The widespread distribution of MAP1A and its interaction with microtubules, actin, and intermediate filaments suggests that it may constitute an important determinant of neuronal and non-neuronal cellular morphology. © 1994 Wiley-Liss, Inc.
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  • 87
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    Cell Motility and the Cytoskeleton 29 (1994), S. 141-154 
    ISSN: 0886-1544
    Keywords: dynein arms ; nexin links ; radial spokes ; relaxation oscillator ; doublet microtubules ; biological oscillators ; computer model ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Ciliary and flagellar motion is driven by the dynein-tubulin interaction between adjacent doublets of the axoneme, and the resulting sliding displacements are converted into axonemal bends that are propagated. When the axoneme is bent in the normal beating plane, force develops across the axoneme in the plane of the bend. This transverse force (t-force) has maximal effect on the interdoublet spacing of outer doublets 2-4 on one side of the axoneme and doublets 7-9 on the opposite side. Episodes of sliding originates as the t-force brings these doublets into closer proximity (allowing dynein bridges to form) and are terminated when these doublets are separated from each other by the t-force. A second factor, the adhesive force of the dynein-tubulin attachments (bridges), also acts to pull neighboring doublets closer together. This force resists termination of a sliding episode once initiated, and acts locally to give the population of dynein bridges a type of excitability. In other words, as bridges form, the probability of nearby bridges attaching is increased by a positive feedback exerted through the interdoublet spacing. A conceptual working hypothesis explaining the behavior of cilia and flagella is proposed based on the above concepts. Additionally, the feasibility of this proposed mechanism is demonstrated using a computer simulation. The simulation uses a Monte Carlo-type algorithm for dynein attachment and adhesive force, together with a geometric evaluation of the t-force on the key microtubule pairs. This model successfully develops spontaneous oscillations from any starting configuration (including a straight position). It is compatible with the physical dimensions, mechanical properties and bridge forces measured in real cilia and flagella. In operation, it exhibits many of the observed actions of cilia and flagella, most notably wave propagation and the ability to produce both cilia-like and flagella-like waveforms. © 1994 Wiley-Liss, Inc.
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  • 88
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    Cell Motility and the Cytoskeleton 29 (1994), S. 195-203 
    ISSN: 0886-1544
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
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  • 89
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    Cell Motility and the Cytoskeleton 29 (1994), S. 339-344 
    ISSN: 0886-1544
    Keywords: microfilament ; phalloidin ; immunoblotting ; immunocytochemistry ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Anti-actin monoclonal antibodies were prepared using phalloidin-stabilized actin that was purified from pea roots by DNase I affinity chromatography. One monoclonal antibody, designated mAb3H11, bound plant actin in preliminary screenings and was further analyzed. Immunoblot analysis showed that this antibody had a high affinity for plant actin in crude and purified preparations but a low affinity for rabbit muscle actin. In immunoblots of plant extracts separated on two-dimensional gels it appeared to bind all actin isoforms recognized by the JLA20 anti-chicken actin antibody. Using immunofluorescent cytochemistry, the antibody was used to observe actin filaments in aldehyde-fixed and methanol-treated tobacco protoplasts. These results indicate that mAb3H11 should be a useful reagent for the study of plant actins. © 1994 Wiley-Liss, Inc.
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  • 90
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    Cell Motility and the Cytoskeleton 29 (1994), S. 366-374 
    ISSN: 0886-1544
    Keywords: cytoskeleton ; microfilaments ; cytochalasin ; gravity ; amiprophos-methyl ; tip-growth ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Apical cells of protonemata of the moss Ceratodon purpureus are unusual among plant cells with sedimentation in that only some amyloplasts sediment and these do not fall completely to the bottom of vertical cells. To determine whether the cytoskeleton restricts plastid sedimentation, the effects of amiprophos-methyl (APM) and cytochalasin D (CD) on plastid position were quantified. APM treatments of 30-60 min increased the plastid sedimentation that is normally seen along the length of untreated or control cells. Longer APM treatments often resulted in more dramatic plastid sedimentation, and in some cases almost all plastids sedimented to the lowermost point in the cell. In contrast, the microfilament inhibitor CD did not affect longitudinal plastid sedimentation compared to untreated cells, although it did disturb or eliminate plastid zonation in the tip. These data suggest that microtubules restrict the sedimentation of plastids along the length of the cell and that microtubules are load-bearing for all the plastids in the apical cell. This demonstrates the importance of the cytoskeleton in maintaining organelle position and cell organization against the force of gravity. © 1994 Wiley-Liss, Inc.
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  • 91
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    Cell Motility and the Cytoskeleton 27 (1994), S. 26-40 
    ISSN: 0886-1544
    Keywords: cleavage furrows ; cytokinesis ; actin ; phalloidin ; myosin ; filamin ; talin ; attachment plaques ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: PtK2 cells of exceptionally large size were microinjected with fluorescently labeled probes for actin, myosin, filamin, and talin in order to follow the assembly of the contractile proteins into the cleavage furrows. Whereas in cells of normal size, there is usually a diffuse pattern of localization of proteins in the cleavage furrow, in these large, flat cells the labeled proteins localized in fibers in the cleavage furrow. Often, the fibers were striated in a pattern comparable to that measured in the stress fibers of the same cell type. The presence of talin in discrete plaques along fibers in the cleavage furrows of the large cells suggests a further similarity between cleavage furrow and stress fiber structure. The presence of filamin in the cleavage furrows also suggests the possibility of an overlapping mechanism in addition to that of a talin mediated mechanism for the attachment of actin filaments to the cell surfaces in the cleavage furrow. A model is presented that emphasizes the interrelationships between stress fibers, myofibrils, and cleavage furrows. © 1994 Wiley-Liss, Inc.
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  • 92
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    Cell Motility and the Cytoskeleton 27 (1994), S. 69-78 
    ISSN: 0886-1544
    Keywords: kinesin ; dynein ; MAP-motor interactions ; microtubule arrays ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Bundles of native microtubules isolated from the ovarioles of hemipteran insects are seen to shimmer when observed using dark-field microscopy. This novel form of microtubule motility becomes even more obvious when the isolated bundles are detergent-extracted and reactivated. We have studied the nucleotide-specificity and the drug-sensitivity of microtubule shimmering in order to obtain information regarding the nature of the motor protein responsible, and to compare its properties with those of previously characterised microtubule motors. The involvement of structural MAPs in the shimmering and in maintenance of microtubule bundles in this system has also been investigated. © 1994 Wiley-Liss, Inc.
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  • 93
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    Cell Motility and the Cytoskeleton 27 (1994), S. 88-96 
    ISSN: 0886-1544
    Keywords: cell movement ; speed ; persistence time ; colcemid ; alveolar macrophage ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The role of microtubules in random cell migration was investigated using time-lapse videomicroscopy to record in vitro the shape and motile behavior of guinea pig alveolar macrophages before and after disrupting microtubules with colcemid. Cell migration was quantified in terms of directional persistence time and speed. Motility was also correlated with morphological polarity: cells having a single lamellipodal region (monopolar cells) migrated, whereas those lacking a lamellipod (apolar cells) or with opposing lamellipodal regions (bipolar cells) did not migrate. Within 2 hours, colcemid caused a shift in polarity from 80% monopolar cells to 40% monopolar and 40% bipolar cells and a corresponding decrease from 80% to 40% in the fraction of migrating cells. Mean persistence time and speed decreased only slightly (approximately 20%) for those cells (still monopolar) which continued to migrate in the presence of colcemid. Persistence time and speed actually increased for many individual cells, indicating that random migration did not require intact microtubules. We conclude that colcemid treatment destabilizes monopolarity, leading to the gradual loss of monopolarity and consequent inhibition of migration. While a cell remains monopolar, it will continue to migrate even in the absence of intact microtubules, but microtubules are required for the long-term maintenance of cellular monopolarity and, thus, for continued motility. © 1994 Wiley-Liss, Inc.
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  • 94
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    Cell Motility and the Cytoskeleton 27 (1994) 
    ISSN: 0886-1544
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 95
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    Cell Motility and the Cytoskeleton 27 (1994), S. 262-271 
    ISSN: 0886-1544
    Keywords: 3T3 cells ; cell motility ; infrared ; phototaxis ; centrosome ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Previous experiments have suggested that 3T3 cells were able to extend pseudopodia toward latex particles up to 60 μm away from the cell body if the particles were irradiated by an infrared beam in the range of 700-900 nm [Albrecht-Buehler, 1991: J. Cell Biol. 114:493-502]. The present article reports that this response of cells to infrared light can be inhibited if the cell center is simultaneously irradiated with a beam of the same light. In marked contrast, the cells responded normally to the presence of infrared light scattering particles if the second beam irradiated other parts of the cell body. The results imply that the cellular mechanism of infrared detection is located at the cell center. The infrared sensing mechanism remains intact in enucleated cells and in cells which were incubated in monensin to vesiculate their Golgi apparatus and inhibit their Golgi functions. Accordingly, it is proposed that the centrosome which contains the centrioles is the only remaining candidate in the cell center for a cellular detection device for the direction of infrared signal sources. The results support an earlier suggestion that centrioles may be such detection devices [Albrecht-Buehler, 1981: Cell Motil. Cytoskeleton 1:237-245]. © 1994 Wiley-Liss, Inc.
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  • 96
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    Cell Motility and the Cytoskeleton 27 (1994), S. 206-218 
    ISSN: 0886-1544
    Keywords: sperm motility ; sperm maturation ; flagella ; protein kinases ; protein kinase inhibitor ; cGMP ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Ejaculated ram sperm were demembranated with Triton X-100, separated from the detergent-soluble matrix, and reactivated [San Agustin and Witman (1993): Cell Motil. Cytoskeleton 24:264-273]. The percent motility of models prepared from freshly washed sperm was comparable to that of the washed sample before demembranation, regardless of whether cAMP was included in the reactivation medium. However, demembranated models derived from aging or metabolically inhibited sperm exhibited a lower percent reactivation and required cAMP to attain the level of motility of freshly washed sperm. Cyclic AMP was ∼100 times more effective than cGMP. The requirement for cAMP could be bypassed by addition of porcine heart cAMP-dependent protein kinase (PKA) catalytic subunit to the reactivation medium, demonstrating that cAMP was acting via PKA. The cAMP stimulation of reactivation was not affected by inclusion of the PKA inhibitor PKI(5-24) in the reactivation medium, but was decreased when the models were preincubated with PKI(5-24) prior to reactivation. The cytosol-free models retained 〉90% of the sperm PKA activity; therefore, the PKA appears to be anchored to internal sperm structures. This PKA could not be extracted by cAMP or Triton X-100 alone, but only by cAMP and Triton X-100 in combination. We conclude that cAMP-dependent protein phosphorylation is critical for sperm motility, but that the essential protein phosphate sites turn over slowly under our reactivation conditions, so that the cAMP requirement is apparent only in models prepared from sperm having a low internal ATP or cAMP content. Interestingly, reactivation was rapidly blocked by the peptide arg-lys-arg-ala-arg-lys-glu, which has been reported to be a selective inhibitor of cGMP-dependent protein kinase. © 1994 Wiley-Liss, Inc.
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  • 97
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    Cell Motility and the Cytoskeleton 27 (1994), S. 286-286 
    ISSN: 0886-1544
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 98
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    Cell Motility and the Cytoskeleton 27 (1994), S. 287-298 
    ISSN: 0886-1544
    Keywords: cilium ; flagellum ; motility ; microtubules ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A physical model developed to explain microtubule sliding patterns in the trypsintreated ciliary axoneme has been extended to investigate the generation of bending moments by microtubules sliding in an axoneme in which the dublets are anchored at one end. With sliding restricted, a bending moment is developed by the polarized shearing interaction between neighbouring doublets, effected by the activity of dynein arms on doublet N pushing N + 1 in a tipward ( + ) direction. In arrested axonemes in which arms on several contiguous doublets are active, the bending moment causes splitting of the 9 + 2 microtubule array into two or more sets of doublets. In the absence of special constraints, splitting depends only on breaking the circumferential interdoublet links most distorted by the bending moment. The analysis, which permits assignment of arm activity to specific microtubules in each of the observed patterns of splitting, indicates that the axoneme will split between doublet N and N + 1 if arms on doublet N are inactive and arms on either N + 1 or N-1 are active. To produce the observed major splits, dynein arms on the microtubules of roughly one-half of the axoneme are predicted to be active, in a manner consistent with the switch-point hypothesis of ciliary motion. Electron microscopic examination indicates that virtually every set of doublets in the split axonemes retains its cylindrical form. Maintenance of cylindrical symmetry can be ascribed to the mechanical properties of the unbroken links, which may resist both tensile and compressive stress, and to active dynein arms. © 1994 Wiley-Liss, Inc.
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    Cell Motility and the Cytoskeleton 27 (1994), S. 361-372 
    ISSN: 0886-1544
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
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    Cell Motility and the Cytoskeleton 28 (1994) 
    ISSN: 0886-1544
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Type of Medium: Electronic Resource
    Library Location Call Number Volume/Issue/Year Availability
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