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  • 2000-2004
  • 1990-1994  (140)
  • 1985-1989  (112)
  • 1992  (140)
  • 1988  (112)
  • Immunohistochemistry  (252)
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  • 2000-2004
  • 1990-1994  (140)
  • 1985-1989  (112)
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  • 101
    ISSN: 1432-0878
    Keywords: In situ hybridization ; Immunohistochemistry ; Corticotropin-releasing factor ; Messenger RNA ; Preoptic nucleus ; Catostomus commersoni (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In situ hybridization procedure with a 32P-labelled synthetic oligonucleotide probe was used to detect corticotropin-releasing factor-encoding messenger RNA (CRF mRNA) in the hypothalamus of the white sucker, Catostomus commersoni. Adjacent sections were immunostained by a sucker CRF-specific antiserum. CRF mRNA-containing neurons were identified by autoradiography in the magnocellular and parvocellular subdivisions of the preoptic nucleus (PON). Many of these neurons were also immunostained by sucker antiserum, showing the same distribution patterns. These results confirm the presence of CRF mRNA and CRF peptide in the white sucker hypothalamus and support the view that the magnocellular and parvocellular neurons of the PON may be involved in the control of adrenocorticotropic hormone secretion from the pituitary in the white sucker.
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  • 102
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 270 (1992), S. 87-93 
    ISSN: 1432-0878
    Keywords: Ovarian nerves ; Development ; Folliculogenesis ; Tyrosine hydroxylase ; Immunohistochemistry ; Electron microscopy ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Sympathetic neurotransmitters have been shown to be present in the ovary of the rat during early postnatal development and to affect steroidogenesis before the ovary becomes responsive to gonadotropins, and before the first primordial follicles are formed. This study was undertaken to determine if development of the ovarian innervation is an event that antedates the initiation of folliculogenesis in the rat, Rattus norvegicus. Serial sections of postnatal ovaries revealed a negligible frequency of follicles 24 h after birth (about 1 primordial follicle per ovary). Twelve hours later there were about 500 follicles per ovary, a number that more than doubled to about 1300 during the subsequent 12 h, indicating that an explosive period of follicular differentiation occurs between the end of postnatal days 1 and 2. Electron microscopy demonstrated that before birth the ovaries are already innervated by fibers containing clear and dense-core vesicles. Immunohistochemistry performed on either fetal (day 19) or newborn (less than 15h after birth) ovaries showed the presence of catecholaminergic nerves, identified by their content of immunoreactive tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine biosynthesis. While some of these fibers innervate blood vessels, others are associated with primordial ovarian cells, thereby suggesting their participation in non-vascular functions. Since prefollicular ovaries are insensitive to gonadotropins, the results suggest that the developing ovary becomes subjected to direct neurogenic influences before it acquires responsiveness to gonadotropins.
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  • 103
    ISSN: 1432-0878
    Keywords: Human cycling endometrium ; Type-2 chain ABO antigens ; Immunohistochemistry ; Genetic and hormonal regulation ; Genetic regulation ; Hormonal regulation ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The blood group ABO(H) determinants are major allogenic antigens in both erythrocytes and tissue of man. These antigens and related carbohydrates are markers of cellular maturation and differentiation in many epithelial tissues and have recently attracted great interest as tumor-associated antigens. Previous studies of endometrial tissues have indicated that glycosylation in this tissue may be related to hormonal stimulation. We have investigated the immunohistochemical distribution of type-2 chain histo-blood group-related carbohydrates in specimens of normal, cycling endometria obtained from hysterectomies on women with known ABO/Lewis erythrocyte type and saliva secretor status. N-acetyllactosamine and Lex were demonstrated to be uninfluenced by the genetic background. A and Aley antigens were exclusively demonstrated in endometria from blood group A individuals, while Ley was expressed in endometria from blood group 0 individuals mainly. The precursor N-acetyllactosamine as well as the terminal H, A, and ALey antigens were shown in only a few cells. In contrast, N-acetyllactosamine substituted by sialic acid and/or fucose residues (Lex, sialosyl-Lex, Ley) were demonstrated in epithelial cells of normal, cycling endometrium, but with both quantitative and qualitative differences in staining relating to the menstrual cycle, indicating that type-2 chain antigens are expressed under both genetic and hormonal influence in human cycling endometrium.
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  • 104
    ISSN: 1432-0878
    Keywords: Cervix ; Uterus ; Eosinophils ; Major basic protein, eosinophil ; Immunohistochemistry ; Parturition ; Rat (Wistar, IFFA-Credo)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Distribution of the eosinophil major basic protein (MBP) was studied in the rat uterus horn and cervix by means of immunohistochemistry using an antiserum raised against rat MBP. Various hormonal contexts were investigated: pre- and post-parturition, the estrous cycle, and ovariectomy followed by hormonal treatment or without treatment. MBP was detectable in the cervix as early as 12 h post-partum, appearing in the stroma close to the myometrium. The MBP had spread throughout the stroma toward the luminal epithelium after a few days. In contrast, no MBP was seen in sections of the corresponding pre- and post-partum uteri and in the pre-partum cervix. In cycling rats, MBP was distributed equally in the cervix and uterus and was more abundant during proestrus and estrus. In ovariectomized rats and in ovariectomized rats subsequently treated with progesterone, no MBP was detected in the cervix or uterus. In the cervix of ovariectomized rats treated with estradiol, MBP first appeared in the muscle layer situated between the two cervical lumina and then reached the stroma; within a few days only the stroma was stained. Inversely, in the uterus MBP-staining first appeared in the stroma. In conclusion, analysis of the distribution of MBP in rat uterus revealed a marked difference in the response of the cervix and horn to a hormonal environment.
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  • 105
    ISSN: 1432-0878
    Keywords: Xenopsin ; Xenopsin precursor fragment ; Immunohistochemistry ; Skin ; Gastrointestinal tract ; Xenopus laevis (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Xenopsin (Xp) and xenopsin precursor fragment (XPF) are bioactive peptides derived from a single precursor molecule; both were isolated previously from extracts of Xenopus laevis skin. The present immunohistochemical study was undertaken to determine the specific cellular localization of these two peptides in the skin and also in the gastrointestinal tract of adult Xenopus. We report here that Xp-like and XPF-like immuno-reactivities co-exist in the granular glands of the skin and specific granular cells in the lower esophagus and stomach. However, only Xp-like immunoreactivity, not XPF-like immunoreactivity, was detected in tall, thin cells of the duodenum and in club-shaped cells of the large intestine. The immunochemical co-localization of the two peptides in specific cells of the skin, lower esophagus and stomach suggests that the same gene is expressed in each of these cells, and that the precursor molecule undergoes similar post-translational processing. In contrast, the observation that certain cells of the duodenum and large intestine display only one peptide immunoreactivity suggests an alternative phenomenon, possibly involving selective peptide accumulation or expression of a different gene.
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  • 106
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 270 (1992), S. 273-279 
    ISSN: 1432-0878
    Keywords: Macrophages ; Small intestine ; Large intestine ; External muscle layer ; Immunohistochemistry ; Histochemistry ; Electron microscopy ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In the external layers of human small and large intestine macrophage-like cells were characterized by immunohistochemical, histochemical and electronmicroscopical methods. Using immunohistochemistry and a number of monoclonal antibodies, the presence and distribution of phenotypic subpopulations of macrophages were evaluated. In all locations macrophage-like cells were identified with antibody EBM11, which recognizes CD68 antigen, C3bi which recognizes CD11b, and partly with an antibody which recognizes protein 150,95 (CD11c). Macrophage-like cells in the external muscle layer were HLA-DR-positive (expressing the MHC class-II antigen), in contrast to macrophage-like cells in the subserosa and submucosa. Macrophage-like cells in the external muscle layer were mostly acid phosphatase-negative, and at the electron-microscopic level they were found to have features of macrophages: primary lysosomes, coated vesicles and pits. However, very few secondary lysosomes were present. Birbeck granules were not observed. It is concluded that in the external muscle layer of human small and large intestine numerous macrophages of a special type are present. It is discussed whether this cell type plays a role in gastrointestinal motility and/or has an immunological function.
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  • 107
    ISSN: 1432-0878
    Keywords: Brain, vertebrate ; Catecholamines ; Tyrosine hydroxylase ; Immunohistochemistry ; Anolis carolinensis (Lacertilia)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using traditional as well as whole-mount immunohistochemistry, we described the location of tyrosine hydroxylase-and dopamine beta hydroxylase-positive cells and fibers in the brain of the lizard Anolis carolinensis. Major catecholaminergic cell groups were in the ependyma in certain ventricular regions, alous coeruleus, anterior hypothalamic and lateral hypothalamic areas, and in the mesencephalic tegmental region, locus coeruleus, nucleus of the solitary tract, vagal motor nucleus, and rhombencephalic reticular formation. Major catecholaminergic fibers, tracts and varicosities included tuberohypophysial, mesolimbic, nigrostriatal, isthmocortical, medullohypothalamic, and coeruleospinal systems. Although the catecholaminergic systems in A. carolinensis are similar to those in the brains of other lizards studied, there are a few species differences. Our information about A. carolinensis will be used to help localize the hypothalamic asymmetry in catecholamine metabolism previously described in this lizard.
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  • 108
    ISSN: 1432-0878
    Keywords: Sensilla ; Immunohistochemistry ; Cryofixation ; Freeze-substitution ; Thermoreceptors ; Hygroreceptors ; Sensory transduction ; Ion pumps ; Antheraea pernyi (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In an attempt to identify and localize the components of voltage sources involved in sensory transduction in insect sensilla, the thermo-/hygrosensitive sensilla of the moth Antheraea pernyi were probed with a polyclonal antiserum against Na+,K+-ATPase in cryofixed and freeze-substituted preparations. The antiserum recognized epitopes on the cytoplasmic membranes of the dendritic inner segments and somata of the sensory cells and also on the cytoplasmic membranes of glial cells surrounding the initial axon segments. The findings support the current concept that ion pumps in the cytoplasmic membranes of the dendritic inner segments and somata of the sensory cells contribute to the maintenance of the resting potential of the sensory cells and to the driving forces generating the receptor currents in response to stimulation of the sensillum. Morphological features and immunohistochemical characteristics of the region of the initial axon segment are also discussed with respect to the initiation of action potentials in these sensilla.
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  • 109
    ISSN: 1432-0878
    Keywords: Degeneration ; Histochemistry ; Immunohistochemistry ; Morphometry ; Muscle ; Stimulation, chronic ; Rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The purpose of this study was to examine the contention that stimulation-induced damage, resulting in degeneration with subsequent regeneration, plays a major role in the transformation of fibre type brought about by chronic electrical stimulation. Data from histological and histochemical sections of 9-day-stimulated rabbit fast-twitch muscles were analysed with multivariate statistical techniques. Fibre degeneration and regeneration varied non-systematically between sample areas at any given cross-sectional level. In the extensor digitorum longus muscle, but not in the tibialis anterior, there was more degeneration in proximal than in distal portions of the muscle. The extensor digitorum longus muscle consistently showed more degeneration than the tibialis anterior muscle. Degeneration was less extensive for an intermittent pattern of stimulation that delivered half the aggregate number of impulses of continuous stimulation. Degeneration and regeneration varied markedly between individual rabbits in each of the groups. Sections that revealed the most degeneration and regeneration also had more fibres that reacted positively with an anti-neonatal antibody. Rigorous analysis of different sources of variation has helped to explain apparent conflicts in the literature. The incidence of muscle fibre damage in the stimulated tibialis anterior muscle is low, showing that the contribution of degenerative-regenerative phenomena to fibre type conversion in this muscle is insignificant.
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  • 110
    ISSN: 1432-0878
    Keywords: Calcitonin gene-related peptide ; Renal pelvis ; Ureter ; Whole-mount preparation ; Immunohistochemistry ; Ureteral ligature ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of calcitonin gene-related peptide-immunoreactive nerve fibers in the renal pelvis and ureter was examined by immunohistochemistry using whole-mount preparations and cryostat sections. The patterns of innervation were contrasted between the pelvis and ureter; the immunoreactive nerve fibers in the pelvis ran parallel to the long axis of each of the circular and longitudinal muscle layers, causing a lattice-like appearance of the nerve fibers. In the ureter, the immunoreactive fibers were accumulated in the subepithelial region and the longitudinal muscle. In both the pelvis and ureter, a portion of the nerve fibers of smaller caliber showed a swollen or beaded structure; they were located in the musculature and beneath the epithelium extending for considerable distances. Ligation of the ureter caused a marked decrease in the immunoreactive nerves in the pelvis and the proximal portion of the ureter, suggesting that the axonal flow in the calcitonin gene-related peptide-containing neurons of the ureter runs towards the pelvis.
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  • 111
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 267 (1992), S. 11-16 
    ISSN: 1432-0878
    Keywords: Kidney ; Distal tubule ; Tamm-Horsfall protein ; Cytokeratin ; Immunohistochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Various antibodies and lectins were used in a histological study of the human renal tubule, particularly of the distal end of the thick limb of the loop of Henle. The thick limb, identified by antibody to Tamm-Horsfall protein, ended abruptly, either at the macula densa or at a variable distance after it. At this point there was an abrupt change in cell size. Confocal microscopy and other techniques showed that this point marked an abrupt beginning of tubular staining by the cytokeratin antibody PKK 2 and the lectin UEA 1, with an abrupt end of staining by the lectin DBA. Distal from this point, there were gradual changes in staining of the tubule by various reagents including other antibodies to cytokeratins. These structural findings suggest that there is a fundamental change in the tubule at the end of the thick limb. The abrupt end to the thick limb in man resembles that seen in the rat and the rabbit.
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  • 112
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 267 (1992), S. 185-192 
    ISSN: 1432-0878
    Keywords: Retina ; Proliferative vitreoretinopathy ; Epiretinal membrane ; Fibronectin ; In situ hybridisation ; Immunohistochemistry ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of fibronectin mRNA and fibronectin in adult human retina and epiretinal membranes was investigated by in situ hybridisation and immunohistochemical techniques. The cells in normal adult retina contained little or no fibronectin mRNA and the retina only showed fibronectin immunoreactivity in retinal vessels. The cells in detached neuroretina did not contain fibronectin message but the vitreoretinal interface of the detached retina exhibited variable fibronectin immunoreactivity. Retinal glia, retinal pigment epithelium and fibroblast-like cells in membranes at the vitreoretinal juncture (epiretinal membranes) showed variable labelling with the fibronectin mRNA probe and all the membranes immunostained for fibronectin. No difference could be detected between membrane cell types in the intensity of labelling with the mRNA probe or for fibronectin immunoreactivity. The results indicate that cells in situ in attached and detached adult human retina do not produce fibronectin. Although fibronectin at the vitreoretinal juncture in retinal detachment is probably partly derived from plasma fibronectin resulting from breakdown of the blood-retinal barrier, ectopic retinal cells produce fibronectin and contribute to the glycoprotein in epiretinal membranes.
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  • 113
    ISSN: 1432-0878
    Keywords: Atrial natriuretic peptides ; mRNA ; Diabetes, type I ; Immunohistochemistry ; Morphometry ; Mouse (NOD)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Atrial natriuretic peptide (ANP) levels in cardiocytes and plasma were examined by using immunohistochemistry, electron microscopy, and radioimmunoassay in non-obese diabetic mice (NOD). Cardiocyte ANP mRNA expression was measured by the polymerase chain reaction method. ANP immunoreactivity in the auricular cardiocytes was more prominent in hyperglycemic mice (NOD-h) than in normoglycemic mice (NOD-n). Ultrastructural examination showed that auricular cardiocytes of the NOD-h group contained more cytoplasmic granules than cells of the NOD-n group. Ultrastructural morphometry indicated that the number of granules per auricular cardiocyte was significantly larger in the NOD-h group than in the NOD-n group. (P〈0.01), whereas the granule diameter was significantly smaller in the NOD-h group (P〈0.01). Radioimmunoassay showed that ANP levels in the NOD-h auricular cardiocytes were significantly higher than those in the NOD-n cardiocytes (P〈0.01); the opposite was true in plasma. Cardiocyte ANP mRNA expression was lower in the NOD-h group than in the NOD-n group.
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  • 114
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 267 (1992), S. 301-306 
    ISSN: 1432-0878
    Keywords: Luteinizing hormone beta-messenger RNA ; In situ hybridization ; Immunohistochemistry ; Pars tuberalis ; Sheep
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The localization of luteinizing hormone beta (LHβ)-mRNA was studied by in situ hybridization in the pars tuberalis of sheep using a homologous sheep double-stranded 32P-or 35S-cDNA. The labelled cDNA probe detected one mRNA sequence in the pars tuberalis by Northern blot analysis; this sequence was similar to that detected in the pituitary. In situ, the labelling of LHβ-mRNA in the horizontal and sagittal tissue sections was found throughout the pars tuberalis. This labelling was prevented by adding an excess of cold probe or treating the sections by ribonuclease before in situ hybridization. Controls showed a labelling in the pars distalis, but not in the median eminence, hypothalamus, cerebral cortex and liver sections. Double labelling by using a specific LHβ-antiserum indicated that the labelling of LHβ-mRNA appeared more intense in LH-containing cells that were found only in the ventral part of the pars tuberalis. These results suggest that the entire pars tuberalis is able to produce the LHβ subunit, but that the level of translation greatly varies according to the location of the cells.
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  • 115
    ISSN: 1432-0878
    Keywords: Mammary gland ; Extracellular matrix ; Menstrual cycle ; Breast cancer ; Immunohistochemistry ; Epithelial cell behaviour ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The normal human mammary gland undergoes a well defined sequence of histological changes in both epithelial and stromal compartments during the menstrual cycle. Studies in vitro have suggested that the extracellular matrix surrounding the individual cells plays a central role in modulating a wide variety of cellular events, including proliferation, differentiation and gene expression. We therefore investigated the distribution of a number of extracellular matrix molecules in the normal breast during the menstrual cycle. By use of indirect immunofluorescence, with specific antibodies, we demonstrated that laminin, heparan sulphate proteoglycan, type IV collagen, type V collagen, chondroitin sulphate and fibronectin undergo changes in distribution during the menstrual cycle, whereas collagen types I, III, VI and VII remain unchanged. These changes were most marked in the basement membrane, sub-basement membrane zone and delimiting layer of fibroblasts surrounding the ductules where basement membrane markers such as laminin, heparan sulphate proteoglycan, and type IV and V collagens appear greatly reduced during the mid-cycle period (days 8 to 22). These results suggest that some extracellular matrix molecules may act as medittors in the hormonal control of the mammary gland, whereas others may have a predominantly structural role.
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  • 116
    ISSN: 1432-0878
    Keywords: Vitamin A deficiency ; Cytokeratins ; Epithelial cells ; Immunohistochemistry ; Rat (BN/BiRij)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Normal epithelial cell differentiation is charactezied by the production of distinct cytokeratin proteins. It is well known that epithelia of several organs show squamous metaplasia in a vitamin A-deficient status. It is not yet known whether these histological changes are concomitant with a change in cytokeratin expression. Therefore, 3-week-old female rats (BN/BiRij) were fed a vitamin A-deficient diet for 8 weeks. The cytokcratin expression in epithelia of various organs was monitored immunohistochemically during the induction of vitamin A deficiency. Therefore, monoclonal antibodies specific for human cytokeratin 4, 5, 5+8, 7, 10, 14, 18 and 19 were used. In a normal vitamin A status, the distributional pattern for the different cytokeratins in rats was similar to that reported for human tissue. No change in cytokeratin expression was seen in trachea, skin, liver and colon at any time point studied. Squamous metaplasia in urinary bladder and salivary glands was observed after six weeks on the vitamin A-deficient diet. This was concomitant with a substitution of cytokeratins 4, 5+8, 7, 18 and 19 by cytokeratin 10. The latter cytokeratin is specific for keratinzed squamous epithelium. A change in cytokeratin expression was observed in bladder, ureter, kidney, salivary glands, uterus and conjunctiva before histological alterations appeared. In conclusion, the changes in cytokeratin expression observed under vitamin A deficiency in epithelia in vivo are in agreement with those described in other studies for epithelial cells in vitro. The changes in cytokeratin expression and the subsequent differentiation into squamous cells occurs in basal cells of the bladder but not in transitional cells. Furthermore, histological alterations are preceded by changes in cytokeratin expression indicating that vitamin A status controls cytokeratin expression in vivo.
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  • 117
    ISSN: 1432-0878
    Keywords: Spleen ; Transplantation ; Immunohistochemistry ; Lymphocytes ; Macrophages ; Stimulation ; Cytokines ; Rat (Lewis) ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Inbred animals (Lewis rats) were used to investigate the regeneration of autologously implanted splenic tissue at intra-omental and subcutaneous sites. Quantitative immunohistology with monoclonal antibodies against lymphocytes and macrophages was performed to analyse the cell density of red pulp (RP), periarteriolar lymphoid sheath (PALS), marginal zone (MZ) and follicle, 7–180 days after transplantation. Antigenic, allogeneic and mitogenic stimulation and Northern blotting were also performed. Transplant groups differed from spleen only in the reduced size of PALS; however, quantitative analysis demonstrated subtle differences between spleen and transplants. The cell density of B-cells and ED-1+ macrophages was reduced in the RP, Tsupp/cyt-cells were decreased and B-cells increased in PALS, and B-cells and Thelper-cells reduced in the MZ. No differences could be detected between the transplant groups. Flow-cytometric analysis of cell suspensions from spleen and transplants revealed a reduction of T-cells (OX-19+), MHC-I and transferrin-receptor-bearing cells in both transplant groups, and a decrease in the number of Thelper-cells and ED-3+ macrophages in subcutaneous transplants. Both transplant groups were defective regarding the allogeneic and pokeweed mitogen response. Aberration of the lipopolysaccharide response was restricted to subcutaneous transplants, which additionally showed abnormal expression of interferon-gamma, interleukin-5 and interleukin-6 mRNA. Thus, subtle alterations of the newly developed microenvironment and/or lymphocyte-homing may influence the regeneration of splenic tissue; the implantation site may represent an important parameter in functional reorganisation.
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  • 118
    ISSN: 1432-0878
    Keywords: Carotid labyrinth ; Ontogeny ; Substance P ; CGRP ; VIP ; Immunohistochemistry ; Rana catesbeiana (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ontogeny of substance P, CGRP (calcitonin gene-related peptide), and VIP (vasoactive intestinal polypeptide) containing nerve fibers in the carotid labyrinth of the bullfrog, Rana catesbeiana, was examined by the peroxidase-antiperoxidase method. The time of appearance of these three peptides was different for each. First, CGRP fibers appeared in the wall of the carotid arch and external carotid arteries, and in a thin septum between these two arteries at an early stage of larval development (stage III). At stage V, substance P immunoreactive fibers appeared, and VIP fibers were detected at the early metamorphic stage (stage XXII). Up to the completion of metamorphosis, the number of these fibers remained low. From 1 to 5 weeks after metamorphosis, substance P, CGRP, and VIP fibers increased in number to varying degrees. By 8 weeks after metamorphosis, the distribution and abundance of these fibers closely resembled those of the adults. Some CGRP and VIP immunoreactive glomus cells were found at the stages immediately before and after the completion of metamorphosis. These findings suggest that substance P, CGRP, and VIP fibers during larval development and metamorphosis may be nonfunctional, and start to participate in vascular regulation only after metamorphosis. The transient CGRP and VIP in some glomus cells may be important for the development of the labyrinth, or may take part in vascular regulation through the close apposition of the glomus and smooth muscle cells (g-s connection).
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  • 119
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 269 (1992), S. 151-158 
    ISSN: 1432-0878
    Keywords: Atrial natriuretic peptide ; Brain natriuretic peptide ; C-type natriuretic peptide ; Heart ; Brain vertebrate ; Immunohistochemistry ; Opsanus beta (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of natriuretic peptide immunoreactivity was determined in the heart and brain of the gulf toadfish Opsanus beta using the avidin-biotin peroxidase technique. Four antisera were used: the first raised against porcine brain natriuretic peptide which cross-reacts with atrial natriuretic and C-type natriuretic peptides (termed natriuretic peptide-like immunoreactivity); the second raised against porcine brain natriuretic peptide which cross-reacts with C-type natriuretic peptide but not with atrial natriuretic peptide (termed porcine brain natriuretic peptide-like immunoreactivity); the third raised against rat atrial natriuretic peptide; and the fourth raised against eel atrial natriuretic peptide. Natriuretic peptide- and porcine brain natriuretic peptide-like immunoreactivity was observed in all cardiac muscle cells of the atrium. In the ventricle, natriuretic peptide-like immunoreactivity was found in all cardiac muscle cells, however porcine brain natriuretic peptidelike immunoreactivity was confined to muscle cells adjacent to the epicardium. There was no discernible difference in the distribution of natriuretic peptide-like immunoreactivity and porcine brain natriuretic peptide-like immunoreactivity in the brain. Immunoreactive perikarya were observed only in the preoptic region of the diencephalon, and many immunoreactive fibres were found in the telencephalon, preoptic area, and rostral hypothalamus, lateral to the thalamic region. There was no immunoreactivity in any region of the hypophysis. A pair of distinct immunoreactive fibre tracts ran caudally from the preoptic area to the thalamic region, from which fibres extended to the posterior commissure, area praetectalis, dorsolateral regions of the midbrain tegmentum, and tectum. Many immunoreactive fibres were present in the rostral regions of the inferior lobes of the hypothalamus and in the dorsolateral and ventrolateral aspects of the rhombencephalon. No immunoreactivity was observed in the heart and brain using rat atrial natriuretic and eel natriuretic peptide antisera. Although the chemical structure of natriuretic peptides in the heart and brain of toadfish is unknown, these observations show that a component of the natriuretic peptide complement is similar to porcine brain natriuretic and/or porcine C-type natriuretic peptides. The presence of natriuretic peptides in the brain suggests that they could be important neuromodulators and/or neurotransmitters.
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  • 120
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 270 (1992), S. 1-6 
    ISSN: 1432-0878
    Keywords: Estradiol receptor ; Endometrium ; Ovariectomy ; Immunohistochemistry ; Ultrastructure ; Pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Serial sections of resting porcine endometrium were analyzed with the monoclonal antibody 13H2 using goat antimouse IgG/5 nm gold as secondary reagent or with either polyclonal antibodies from goat #402 or the rat monoclonal antibody H222, both in combination with protein G/12 nm gold. A modestly higher labelling of nuclei than of cytoplasm was seen only with the monoclonal antibody H222. Polyclonal #402 and monoclonal 13H2 showed fewer attachments over nuclear than over cytoplasmic areas. The highest densities of attachment and of predominantly cytoplasmic labelling were obtained with the monoclonal antibody 13H2. The results confirm the earlier assumption of a restricted accessiblity of estradiol receptor in the cytoplasm of resting cells for immunoreagents.
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  • 121
    ISSN: 1432-0878
    Keywords: Invertebrate immunity ; Coelomocytes ; Encapsulation ; Melanin ; Cytochemistry ; Immunohistochemistry ; Nereis diversicolor (Annelida)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary We attempted to identify the nature and origin of the pigment produced by the marine worm Nereis diversicolor in order to isolate, in inert brown capsules, foreign objects introduced into its body cavity. This brown pigment, characterized by cytochemical techniques, could be a melanin. The activity of the enzyme phenoloxidase responsible for melanin biosynthesis was detected by enzyme cytochemistry techniques in vacuoles and the Golgi apparatus of coelomocytes activated by the presence of foreign bodies. Morphological techniques combined with a monoclonal immunological probe enabled us to establish that the “G2” granulocytes contain both the precursor of the pigment in dense bodies and the capacity for phenoloxidase synthesis when activated to encapsulate foreign bodies. The “G2” granulocyte may therefore be compared to a melanocyte in which melanin is not stored as in mammals, but immediately extruded following synthesis in the form of a thick fluid.
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  • 122
    ISSN: 1437-1596
    Keywords: Fibronectin ; Immunohistochemistry ; Wound age
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Law
    Description / Table of Contents: Zusammenfassung Es wurden 53 vitale Hautwunden mit einem Wundalter von wenigen Sekunden/Minuten
    Notes: Summary We analyzed the distribution of fibronectin in routinely embedded tissue specimens from 53 skin wounds and 6 postmortem wounds. In postmortem wounds a faint but focal positive staining was exclusively found at the margin of the specimens which dit not extend into the adjacent stroma. Vital wounds were classified into 3 groups. The first comprising lesions with wound ages ranging from a few seconds to 30 min, the second comprising those with wound ages upt to 3 weeks, and the third group with lesions more than 3 weeks old. Ten out of 17 lesions with a wound age up to 30 min showed a clear positive reaction within the wound area. Three specimens in this group were completely negative, while in 4 additional cases the result was not significantly different from postmortem lesions. These 7 cases were characterized by acute death with extremely short survival times (only seconds). In wounds up to 3 weeks old fibronectin formed a distinct network containing an increasing number of inflammatory cells corresponding to the wound age. In 2 cases with a survival time of 17 days and in all wounds older than 3 weeks fibronectin was restricted to the surface of fibroblasts and to parallel arranged fibers in the granulation tissue without any network structures. We present evidence that fibronectin is a useful marker for vital wounds with a survival time of more than a few minutes. Fibronectin appears before neutrophilic granulocytes migrate into the wound area. Since a faint positive fibronectin staining is seen in postmortem lesions and bleedings, we propose that only those wounds which show strong positive fibronectin staining also extending into the adjacent stroma should be regarded as vital.
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  • 123
    ISSN: 1437-1596
    Keywords: Immunohistochemistry ; ABH-related antigens ; Human male genital tract ; Immunohistochemistry ; Antigene des ABH-Komplexes ; Männlicher Genitaltrakt
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Law
    Description / Table of Contents: Zusammenfassung Die Lokalisation (and Verteilung) der Antigene des ABH-Komplexes im Bereich der inneren Geschlechtsorgane des Mannes wurde mittels monoklonaler Antikörper unter Benutzung einer Avidin-Biotin Technik untersucht. Dabei konnten positive Reaktionen im Hoden und im Ductus epididymidis lediglich an Erythrozyten und Endothelzellen beobachtet werden. Die Expression von ABH-Antigenen in den Ductuli efferentes testis, im Ductus epididymidis, in den Samenbläschen und der Prostata wird offensichtlich komplex durch H-, Se-, Le- und X-Gene kodiert. Die Resultate der vorliegenden Untersuchungen zeigen, daß die ABH-Antigene der Spermienoberfläche offensichtlich aus der Samenflüssigkeit stammen und die ABO-, H-, Se-, Le-und X-Gene gewebsabhängig unterschiedlich exprimiert werden.
    Notes: Summary The localization of ABH related antigens in human male reproductive tract was examined using monoclonal antibodies and an avidin biotin complex method. No positive reaction with blood group antibodies on spermatozoa was observed in testis and ductus epididymidis apart from erythrocytes and endothelial cells. The expression of ABH and ABH related antigens in ductuli efferentes testis, ductus epididymidis, seminal vesicle and prostate was complexly coded by a combination of H, Se, Le and X genes. The results obtained in this study indicate that the ABH antigens detected on spermatozoa of seminal stains are coating antigens and not inherent to the cell membrane, and the ABO, H, Se, Le and X genes are subjected to a tissue-dependent differential expression.
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  • 124
    ISSN: 1437-1596
    Keywords: Immunohistochemistry ; Collagen IV and VII ; Basement Membrane ; Wound Age ; Immunhistochemie ; Kollagen IV ; Kollagen VII ; Basalmembran ; Wundalter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Law
    Description / Table of Contents: Zusammenfasssung Es wurden 62 menschliche Hautwunden (Operationsnähte, chirurgisch versorgte Stich-und Riß-Quetsch-Wunden) untersucht. Neben Kollagen IV wurde in 27 Fällen zusätzlich Kollagen VII immunhistochemisch dargestellt. Es zeigte sich hierbei eine weitgehende Co-Verteilung von Kollagen IV und VII im Wundgebiet ohne daß relevante wundaltersabhängige Unterschiede bezüglich der Lokalisation im Bereich des Epithel-Defektes feststellbar waren. Basalmembran-Fragmente traten erstmals in 4 Tage alten Hautwunden auf. Frühestens 8 Tage nach Verletzung fanden wir eine komplette epidermale Basalmembran. Dies war in allen Präparaten mit einem Wundalter über 21 Tagen der Fall. Der Zeitraum zwischen dem B. und 21. Tag nach Wundsetzung war charakterisiert durch eine erhebliche Variabilität der Befunde mit teils kompletter, teils fragmentiert vorliegender, teils auch noch vollständig fehlender Basalmembran im Defekt-Bereich.
    Notes: Summary In 62 human skin wounds (surgical wounds, stab wounds and lacerations after surgical treatment) we analyzed the immunohistochemical localization of collagen IV in the epithelial basement membrane. In 27 of these wounds the distribution of collagen VII, which represents a specific component of the basement membrane of stratified epithelia, was also analyzed. We were able to demonstrate a virtually identical co-distribution of both collagen IV and VII in the wound area with no significant time-dependent differences in the appearance of both collagen types. Fragments of the epithelial basement membrane could be detected in the wound area from as early as 4 days after wounding and after 8 days a complete restitution of the epithelial basement membrane was observed. In all cases with a wound age of more than 21 days the basement membrane was completely reformed over the former lesional area. The period between 8 and 21 days after wounding was characterized by a wide variability ranging from complete restitution to deposition of basement membrane fragments or total lack of the epidermal basement membrane.
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  • 125
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    International journal of legal medicine 105 (1992), S. 99-103 
    ISSN: 1437-1596
    Keywords: Myofibroblasts ; Alpha-smooth muscle actin ; Desmin ; Immunohistochemistry ; Wound age ; Myofibroblasten ; Alpha-Aktin ; Desmin ; Immunhistochemie ; Wundalter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Law
    Description / Table of Contents: Zusammenfassung Es wurden 66 menschliche Hautwunden mit einem Wundalter zwischen 20 Stunden und 7 Monaten sowie komplikationsloser Wundheilung ausgewertet. Nach immunhistochemischer Darstellung von alpha-Aktin und Desmin wurde das zeitabhängige Auftreten positiv reagierender Myofibroblasten im Wundgebiet untersucht. Es zeigte sich hierbei, daß in Hautwunden mit einem Wundalter unter 5 Tagen keine positiv anfärbbaren Zellen zu beobachten waren. In 57% (25 von 44 Fällen) der Hautverletzungen, die zwischen 5 und 31 Tagen überlebt worden waren, fanden sich im Granulationsgewebe alpha-Aktin haltige Myofibroblasten. Besonders zahlreiche, positiv reagierende Zellen traten zwischen ca. 16 bis 31 Tagen nach Wundsetzung auf, konnten jedoch auch bereits in Hautwunden jüngeren Alters beobachtet werden. In 2 von 7 Fällen mit einem Wundalter zwischen 1 und 7 Monaten (29%) liesen sich ebenfalls alpha-Aktin positive Myofibroblasten im Wundgebiet nachweisen. Desmin-haltige Myofibroblasten konnten nicht beobachtet werden. Die Ergebnisse zeigen, daß alpha-Aktin positive Myofibroblasten bereits mit Ausbildung typischen Granulationsgewebes ab ca. dem 5. Tag nach Verletzung im Wundgebiet auftreten. Der Nachweis positiv reagierender Zellen im Wundgebiet läßt jedoch aufgrund der Variabilität der Befunde keine weitere Differenzierung des Wundalters zu. Da alpha-Aktin-positive Myofibroblasten im Untersuchungsgut auch noch in einer Hautwunde mit einem Alter von 2 Monaten und 13 Tagen beobachtet werden konnten, ist die im Tierexperiment gefundene maximale Nachweisbarkeitsdauer von 30 Tagen auf das Granulationsgewebe menschlicher Hautwunden nicht übertragbar.
    Notes: Summary Human skin wounds (66) inflicted between 20 h and 7 months prior to biopsy were studied. In order to identify the type of cellular differentiation of the fibroblastic cells in the granulation tissue, alpha-smooth muscle actin and desmin were immunohistochemically localized. The value of any presumed time-dependent appearance and/or disappearance of positively stained cells was tested for the estimation of wound age. In skin specimens with a wound age less than 5 days (n =15) no typical granulation tissue had developed and no alpha-actin-positive myofibroblasts could be detected. The first appearance of positively reacting myofibroblasts was noted in a 5-day-old wound. In 57% of the lesions with a wound age between 5 and 31 days (25 out of 44 cases) typical granulation tissue formation was present and myofibroblasts with positive reaction for alpha-smooth muscle actin could be identified. Numerous positively reacting cells could generally be found in wounds aged between 16 and 31 days, but also in wounds less than 16 days old. In 29% of the cases with a wound age of more than 31 days (2 out of 7 cases) alpha-sma-positive myofibroblasts also occured. Fibroblastic cells positive for desmin could not be seen at all in our series. Our results demonstrate the appearance of alpha-sma-positive myofibroblasts with the initial formation of typical granulation tissue in human skin lesions as early as approximately 5 days after wounding. In contrast to recent experimental results these cells remained detectable in wounds aged more than 2 months in some cases. The immunohistochemical detection of actin-positive cells, therefore, demonstrates whether an unknown skin wound is aged approximately 5 days or more. Even though a time-dependent decrease of myofibroblasts in human granulation tissue after 31 days in human wounds seems probable, the extended presence (up to about 2 months) of these cells allows no further exact age determination of older wounds.
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  • 126
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    European archives of oto-rhino-laryngology and head & neck 249 (1992), S. 24-27 
    ISSN: 1434-4726
    Keywords: Immune-mediated otitis media ; T-cell subsets ; Immunohistochemistry ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary To examine the role of T-cell subsets in immune-mediated otitis media with effusion induced by keyhole limpet hemocyanin (KLH), we used immuno-histochemical methods to investigate the kinetics of immunocytes of the middle ear (ME) and eustachian tube (ET) in healthy BALB/c mice. Antibodies against murine macrophages and granulocytes (anti-Mac-1), helper T cells (anti-Lyt-1), suppressor T cells (anti-Lyt-2), immunoglobulins (anti-IgG, -IgM, -IgA), secretory component (SC) and KLH were used. The ME exhibited a substantial immune response, whereas the response of the ET was minor and was associated with a secondary ME immune response. After KLH challenge, an effusion with an extensive infiltration of inflammatory cells (Mac-1, IgG+ and IgM+ cells) was observed at days 1 and 3 in the ME cavity and rapidly disappeared by day 7. Within the ME mucosa, a large number of cells was observed at days 1 and 3, peaking on day 7 when a submucosal lymphoid infiltration was detected. In the immune response of the ME mucosa, Mac-1 cells were the predominant cell type followed by helper T cells, IgG+ cells, IgA+ cells and then IgM+ cells. Suppressor T cells were rarely detected after KLH challenge. SC was present within ME epithelial cells from days 1 to 14. From these findings, we conclude (1) that the majority of infiltrating cells in the ME cavity originate from circulating blood; (2) that the ME mucosa has an excellent capacity to mount a strong immune response, including mucosal immunity, through the accumulation of immunocytes for antigen processing and antibody production; (3) that elimination of antigen appears to be the most important factor for returning the immune response to a quiescent state.
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  • 127
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    European archives of oto-rhino-laryngology and head & neck 249 (1992), S. 40-43 
    ISSN: 1434-4726
    Keywords: Immunohistochemistry ; Tyrosine hydroxylase ; Sympathetic nerve ; Larynx-Dog
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The sympathetic innervation of the canine larynx was investigated using tyrosine hydroxylase (TH) immunohistochemistry. Many tyrosine hydroxylase immunoreactive (TH-IR) nerve fibers were observed around arteries and arterioles in the laryngeal mucosa and intrinsic laryngeal muscles. In the glandular region, TH-IR fibers were also found, with some of these fibers terminating around the basement membranes of the glandular cells. The quantity of TH-IR fibers in the mucosa differed among regions of the larynx. Many of these fibers could be found in the laryngeal surface of the epiglottis as well as the posterior glottis. These findings suggest that TH-IR fibers may directly innervate muscles in the intrinsic larynx.
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  • 128
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    European archives of oto-rhino-laryngology and head & neck 249 (1992), S. 52-55 
    ISSN: 1434-4726
    Keywords: Calcitonin gene-related peptide ; Canine larynx ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Immunohistochemistry was used to investigate the distribution pattern of calcitonin gene-related peptide (CGRP) nerve fibers in the laryngeal mucosa, glands and intrinsic muscles of the dog. CGRP immunoreactive nerve fibers were found more frequently than substance P immunoreative nerve fibers in every region of the larynx. In the epithelia, CGRP nerve fibers were mainly found in the epiglottis, arytenoid region and subglottis. Many taste buds were observed in the arytenoid region and were densely innervated by the CGRP nerve fibers. In the lamina propria, the plexus of CGRP nerve fibers was present, with some of these fibers associated with blood vessels. Laryngeal glands were also innervated by a few CGRP nerve fibers. In the intrinsic laryngeal muscles, abundant immunoreactivity was observed and many motor end-plate-like structures were found with CGRP immunoreactivity. These findings strongly suggest that CGRP plays an important role in all of the sensory, motor and autonomic nervous systems of the larynx.
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  • 129
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    European archives of oto-rhino-laryngology and head & neck 249 (1992), S. 393-399 
    ISSN: 1434-4726
    Keywords: Olfactory epithelia ; Olfactory disorder ; Immunohistochemistry ; Classification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary We have previously demonstrated that human olfactory epithelia can be classified into five grades according to the degree of degeneration present in patients with various kinds of olfactory disorders. In practice, however, the occurrence of additional types of cell changes in other kinds of olfactory disorders and findings with immunohistochemical techniques have led us to re-evaluate our previous classification. In the present study, changes in olfactory epithelia from ten patients with various kinds of olfactory disorders are discussed and a revised classification is proposed. Microvillar and differentiating cells were also evaluated in the epithelium studied.
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  • 130
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    European archives of oto-rhino-laryngology and head & neck 249 (1992), S. 87-90 
    ISSN: 1434-4726
    Keywords: Cholesteatomas ; Macrophages ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Since a heavy cellular infiltrate is seen in the stroma of most aural cholesteatomas, we attempted to characterize this cell population in more detail using monocyte/macrophage-specific monoclonal antibodies. KiM1+ (specific for CD 11c antigen, the 150kDa α-chain of a leukocyte integrin), and KiM6+ phagocytes were present in two- or fourfold higher numbers in the stroma of the six excised cholesteatomas than in the control tissues. Since the stroma of the cholesteatoma is devoid of microvessles, the typical perivascular localization of dermal macrophages was not seen in the cholesteatomas studied. The density of the macrophages in the normal ear skin was much higher in the upper dermis than in the lower dermis. In the cholesteatomatous specimens, the phagocytes were evenly scattered within the connective tissue and the cellular infiltrate. In contrast to diseased skin, no Mac 387+ macrophages were detected in the cholesteatomas. A great number of phagocytic cells closely resembling dermal macrophages was found in the stroma of the cholesteatomas and probably contributes to an active autoimmune process.
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  • 131
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    European archives of oto-rhino-laryngology and head & neck 249 (1992), S. 243-247 
    ISSN: 1434-4726
    Keywords: Epidermal growth factor receptor expression ; Normal oral mucosa ; Dysplastic epithelia ; Squamous cell carcinomas ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The expression of the receptor for epidermal growth factor (EGF) has been determined on oral squamous cell carcinomas. Immunoreactive receptor was localized using a monoclonal anti-EGF-receptor antibody which reacts with sequences in the external domain of the receptor. Frozen sections were studied from 40 patients with squamous cell carcinomas. In 16 sections from the patients with the squamous cell carcinomas, normal differentiated oral mucosa was included and in 7 of these the patients had received preoperative radiotherapy. Sections from 6 other patients with squamous cell carcinoma contained dysplastic epithelia. EGF-receptor-positive cells were present in the basal cell layer on normal differentiated oral mucosa. In sections from patients receiving preoperative radiotherapy the EGF-receptor-positive cells were also found in the spinous cells. In dysplastic epithelia nearly all cells stained for the receptor. The distribution and staining intensity of the EGF receptor varied in the oral squamous cell carcinomas, 36 were positive. The staining pattern in the carcinomas obtained from patients receiving preoperative radiotherapy was not altered qualitatively. Nearly all poorly differentiated cells were stained, but when the tumor was moderately to well differentiated a reduction in the extent of staining in certain areas was seen, paralleling the findings observed in the differentiated upper layers of the normal oral mucosa. This was most pronounced for the epithelial pearls, where the EGF-receptor-positive cells were localized to the undifferentiated cells in the periphery. The results of the present investigation confirm the presence of the EGF receptor on undifferentiated cells, with the extent of the staining reaction on oral squamous cell carcinomas varying inversely with cellular differentiation.
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  • 132
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    European archives of oto-rhino-laryngology and head & neck 249 (1992), S. 385-388 
    ISSN: 1434-4726
    Keywords: Regeneration ; Recurrent laryngeal nerve ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The recurrent laryngeal nerve (RLN) consists of various motor, sensory and autonomic nerve fibers, although it has not been established whether different neuronal types exhibit a similar ability to regenerate. To address this question, freezing was used to injure the cat RLN fibers and the presence or absence of immunoreactivity for neuropeptides or transmitter-synthesizing enzymes was then examined as a marker to classify the fibers. In the control RLN, calcitonin gene-related peptide-immunoreactive (CGRP-IR) fibers were the highest in number and were distributed throughout the nerve fascicles. The number of substance P-immunoreactive (SP-IR) fibers was about 40% that of CGRP-IR fibers, while a portion of CGRP-IR fibers was found to contain SP immunoreactivity. Relatively low numbers of tyrosine hydroxylase-immunoreactive (TH-IR) and neuropeptide Y (NPY-IR) nerve fibers were seen which tended to form clusters. The distribution pattern of NPY-IR fibers was very similar to that of TH-IR fibers. In the regenerating RLN 1 week after the freezing injury, the fastest growing axons were CGRP-IR, while the regenerating rates of SP-IR, TH-IR and NPY-IR fibers were slower than that of CGRP-IR fibers. These results suggest that the ability for neurite regeneration varies among neuron types and that CGRP-IR fibers possess the most rapid ability to regenerate.
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  • 133
    ISSN: 1432-136X
    Keywords: Stanniocalcin purification ; Amino acid sequencing ; Immunohistochemistry ; Intestinal Ca2+ influx ; Atlantic cod, Gadus morhua
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Chum salmon (Oncorhynchus keta) stanniocalcin was purified, partially identified and tested for bioactivity in an assay on the intestinal calcium uptake in a marine teleost (Gadus morhua). Basic ethanol extraction, ion exchange chromatography, gel filtration and reverse-phase high-performance liquid chromatography resulted in the isolation of a homogenous glycoprotein that appears as a 46-kDa product under non-reducing conditions and as a 23-kDa product under reducing conditions after sodium dodecylsulphate-polyacrylamide gel electrophoresis. The glycoprotein is likely to be a homodimer composed of two subunits of 23 kDa each. Further characterization indicates homology to Australian eel, sockeye salmon, coho salmon and rainbow trout stanniocalcin, and the glycoprotein is thus concluded to be stanniocalcin. Stanniocalcin-like immunoreactivity was demonstrated in the corpuscles of Stannius of the Atlantic cod, with a specific antiserum raised against purified chum salmon stanniocalcin. The physiological importance and the biological activity of chum salmon stanniocalcin was tested by evaluating its effect on intestinal calcium uptake by the Atlantic cod in vitro. The intestine was perfused, both vascularly and through the intestinal lumen, and the calcium mucosa-to-serosa flux was measured using 45Ca2+ as a tracer. Stanniocalcin decreased the intestinal calcium uptake in a dose-related manner by 13.5% and 22.4% at doses of 2.2 and 10.9 nM stanniocalcin, respectively. The results establish the intestine as a target organ for stanniocalcin in marine teleosts.
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  • 134
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    The @Anatomical Record 234 (1992), S. 443-451 
    ISSN: 0003-276X
    Keywords: p21ras ; ras ; Immunohistochemistry ; ABC ; Endoderm ; Rat embryo ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Expression of ras cellular oncogenes during the early postimplantation period in the rat was investigated using immunohistochemistry to p21ras was used in an indirect avidin-biotin-peroxidase (ABC) technique. Positive staining indicating the presence of p21ras was found in embryos from 6.5 to 12 days embryonic age. In early egg cylinders (6.5 days), positive staining for p21ras was observed on the ectoplacental cone, primitive ectoderm and trophectoderm, while primitive endoderm and parietal endoderm appeared paler. In later egg cylinder stages (7.5 days), strong positive staining was observed in the primitive embryonic ectoderm and ectoplacental cone, but parietal and visceral endoderm still appeared to be devoid of positive staining. As development proceeded during primitive streak stages, the visceral and parietal endoderm became positively stained. By 10 days, all tissues appeared to be positive for p21ras, with strong staining appearing in the heart and neural elements. Therefore, p21ras does not appear to be ubiquitous in the rat conceptus prior to gastrulation, but shows differential distribution, appearing later in endodermal derivatives. Possibly p21ras is involved in determination of the ectodermal and endodermal lineages.© Willey-Liss, Inc.
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  • 135
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    The @Anatomical Record 234 (1992), S. 549-554 
    ISSN: 0003-276X
    Keywords: Melatonin ; Harderian gland ; Immunohistochemistry ; Mink (Mustela vison) ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: In the Harderian gland of the female mink, either intact or killed after a bilateral ablation of the cervical superior ganglion, almost all of the cells of the alveoli were immunolabelled with anti-melatonin antiserum. Animals were killed during the day or during the night. The immunolabelling was observed only in the cytoplasm, while the nucleus remained unstained. Using successive dilutions of the antiserum on serial sections of the Harderian gland to qualitate the melatonin content, a circadian rhythm of melatonin immunoreactivity was observed. The intensity of immunofluorescence labelling was higher in intact animals killed during the day than in those killed during the night. These results could be explained by the inhibitory or stimulatory influence of pineal melatonin released during the night on melatonin synthesis or release in the Harderian gland, respectively. In the Harderian gland of ganglionectomized animals, the intensity of melatonin immunofluorescence was lower than in intact animals killed during the day. It is concluded that the Harderian gland might be involved in the perception of the day/night cycle and that melatonin synthesis/secretion was likely controlled by the cervical superior ganglion in this organ.© Willey-Liss, Inc.
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  • 136
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    The @Anatomical Record 234 (1992), S. 587-592 
    ISSN: 0003-276X
    Keywords: Dental pulp ; Intermediate filaments ; Actin filaments ; Immunohistochemistry ; Development ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The localization of different cytoskeletal proteins (keratin, vimentin, desmin, actin, and α-smooth muscle actin) was examined by immunohistochemistry in normal human adult dental pulp and compared with dental papilla of tooth germs. Keratin and actin were localized in enamel organ. Vimentin and actin were observed in the dental papilla and in the adult dental pulp. Desmin and α-smooth muscle actin were present only in the vessel walls. These data are discussed paying particular attention to the origin and the peculiar functional characters of the dental papilla and pulp.© Willey-Liss, Inc.
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  • 137
    ISSN: 0003-276X
    Keywords: Dogs ; Immunohistochemistry ; Anal region ; Intermediate filaments ; Keratin ; Vimentin ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The intermediate filament labeling pattern of the epithelial structures of the canine anal region was studied with different polypeptide specific keratin monoclonal antibodies (MoAbs) and with a monoclonal and polyclonal vimentin antibody. The epithelial structures in this region could be discriminated and characterized by differences in their keratin staining pattern. The basal cells in the different epithelial structures showed a similar staining pattern characterized by reactivity with MoAbs staining keratins 5, 8, 14, and 17. Columnar epithelial cells showed a completely different phenotype mostly characterized by reactivity with MoAbs staining keratins 7, 5, 8, 18, and 19. A restricted number of differentiated perianal gland cells showed perinuclear vimentin staining. Myoepithelial cells did not stain for vimentin, but, as other basal cells, were positive for MoAbs staining keratins 5, 8, 14, and 17.© Willey-Liss, Inc.
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  • 138
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    Molecular Reproduction and Development 33 (1992), S. 165-171 
    ISSN: 1040-452X
    Keywords: Boar spermatozoa ; Immunohistochemistry ; Monoclonal antibody ; Seminal plasma ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Sperm coating proteins of 16, 17, and 19 kDa have been purified from boar seminal plasma. The 17 kDa protein has been identified as an antigen recognized by monoclonal antibody ACR.3 and is thus identical to low molecular mass zona pellucida binding protein from boar spermatozoa (Moos et al., 1990). The 17 and 19 kDa proteins are glycosylated and tend to form hetero-complexes. The 17 kDa ACR.3 antigen is sequentially released from the sperm cell surface during capacitation and, after induction of the acrosome reaction, the 16 kDa form was also observed. Immunocytochemical studies on boar reproductive tissues have suggested that the seminal vesicle epithelium may be the source of these proteins. © 1992 Wiley-Liss, Inc.
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    Molecular Reproduction and Development 32 (1992), S. 62-66 
    ISSN: 1040-452X
    Keywords: Mos ; Immunohistochemistry ; Western blot ; Basement membrane ; Stereocilia ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: An anti-Mos protein monoclonal antibody, 4A6, was used to investigate the distribution of the antigen in the epididymis, in which the c-mos gene is reportedly expressed. The 4A6-reactive antigen was found on the basement membrane and luminal surface of the epithelial cells in the caput epididymis of BALB/c male mice as well as in the proximal corpus epididymis, the cauda epididymis, and the vas deferens. The 4A6 antigen was also found on the luminal surface of the epithelial cells in the epididymis of male germ cell-deficient C57BL/6J-Wv/Wv mice. This confirmed that the 4A6 antigen does not derive entirely from the testicular c-Mos protein but is synthesized in the epididymis. Western blot analysis revealed that the molecular weight of the epididymal 4A6 antigen was 50 kDa, which is unusually high for the c-Mos protein. With its specific distribution in the epididymis, the protein should play a specific role in functions of the epididymis. © 1992 Wiley-Liss, Inc.
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  • 140
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    American Journal of Anatomy 193 (1992), S. 300-313 
    ISSN: 0002-9106
    Keywords: Shell-less chick embryo culture ; In situ hybridization ; Immunohistochemistry ; TGF-β ; Extracellular matrix ; Mineralization ; Bone development ; Intramembranous ossification ; Chondrogenesis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The developmental process of intramembranous ossification involves bone formation directly from mesenchymal differentiation without a cartilage intermediate. We have previously observed that systemic calcium deficiency in the developing chick embryo, produced by long-term shell-less culture, results in the appearance of chondrocyte-like cells in the calvarium, a parietal bone which normally develops via intramembranous ossification. This investigation aims to analyze the mechanism underlying this calcium deficiency-related, aberrant appearance of cartilage phenotype in the chick embryonic calvarium. In view of the reported involvement of transforming growth factor β (TGF-β) in osteogenesis and chondrogenesis, we have examined and compared here the expression of TGF-β in the chick embryonic calvaria of normal (in ovo development, NL), shell-less (SL), and calcium-supplemented SL (SL + Ca) embryos. TGF-β expression was analyzed at the mRNA level by blot and in situ cDNA hybridization, and at the protein level by immunohistochemistry and immunoblotting. The results presented here indicate that: (1) TGF-β is expressed in the chick embryonic calvarium by both periosteal cells and osteocytes, as revealed by in situ hybridization and immunohistochemistry; (2) TGF-β expression is significantly increased in SL calvarium compared to NL calvarium, at both protein and mRNA levels; (3) the number of TGF-β expressing cells increases in the SL calvarium, particularly along the central, subcambial core region of the bone; and (4) exogenous calcium repletion to the SL embryo affects the expression of TGF-β such that the pattern approaches that in the NL embryo. Taken together, these results indicate that altered TGF-β expression accompanies the aberrant appearance of cartilage phenotype caused by systemic calcium deficiency. We postulate that normal cellular differentiation along the osteogenic pathway during embryonic intramembranous ossification is crucially dependent on regulated TGF-β expression. © 1992 Wiley-Liss, Inc.
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  • 141
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    Surgical and radiologic anatomy 10 (1988), S. 137-144 
    ISSN: 1279-8517
    Keywords: Retina ; Dopamine ; Immunohistochemistry ; Parkinson's disease
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Résumé Les nouvelles techniques morpho-fonctionnelles d'étude des cellules nerveuses, telles que l'injection intra-cellulaire et la mise en évidence immunohistochimique des neurotransmetteurs, permettent une nouvelle approche de l'architecture fonctionnelle des circuits rétiniens. Deux types de cellules dopaminergiques sont décrits: les cellules amacrines et les cellules interplexiformes. Ces dernières, qui possèdent des prolongements à la fois dans les couches plexiformes interne et externe, forment un circuit récurrent capable d'agir sur le couplage des cellules horizontales. Deux substances localisées dans de telles cellules, la dopamine et le GABA, ont une action antagoniste sur le couplage des cellules horizontales et contrôlent la taille de leurs champs récepteurs qui intervient dans le codage des contrastes. Des modifications de l'ERG, des PEVs et de la sensibilité au contraste sont enregistrés chez les Parkinsoniens. Ces modifications étant identiques à celles observées chez les modèles animaux dont le système dopaminergique rétinien a été détruit, suggèrent une dégénérescence de ce système dans la maladie de Parkinson. L'étude des neurones dopaminergiques, mis en évidence par immunohistochimie de la tyrosine hydroxylase, dans la rétine de 5 malades, a permis d'observer une diminution de l'innervation dopaminergique dans la rétine centrale des Parkinsoniens.
    Notes: Summary Recent morphologic and functional techniques for the study of nerve cells, such as intracellular injection and neurotransmitter immunohistochemistry, allow a new approach to the functional architecture of the retinal circuitry. Two types of dopaminergic cells are described: amacrine cells and interplexiform cells. These latter cells, which send processes to both the inner and outer plexiform layers, form a feedback loop acting the level of horizontal cell coupling. Two molecules localized in such cells, dopamine and GABA, have antagonistic effects on horizontal cell coupling and regulate the diameter of their receptive fields which code for contrast. Changes in the ERG, VEPs and contrast sensitivity occur in Parkinsonian patients and are identical to those observed in animal models whose dopaminergic retinal system has been destroyed, thus suggesting a degenerative process of this system in Parkinson's disease. The observation of dopamine neurons, labelled by their tyrosine hydroxylase immunoreactivity, in the retina of 5 patients, led to the observation of reduced dopamine innervation in the central retina of Parkinsonian patients.
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  • 142
    ISSN: 1432-1440
    Keywords: Thyroid lymphoma ; B-CLL ; Centroblastic lymphoma ; Immunohistochemistry ; Gene rearrangement analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary A 67-year-old woman suffering since 5 years from a so far nontreated B-CLL underwent hemithyroidectomy for a rapidly enlarging tumor. Histologically, the coincidence of a centroblastic lymphoma and thyroidal infiltration by the CLL was diagnosed. Immunohistology revealed typical immunoprofils for both, B-CLL and centroblastic lymphoma on the background of B cell differentiation antigens. The bitypical immunoglobulin light chain expression — λ on the B-CLL cells and κ on the centroblasts — suggested biclonality. This was confirmed by gene rearrangement analysis of peripheral leukemia cells and tumor tissue. Thus, the final diagnosis of a primary thyroidal lymphoma of the centroblastic type (stage IE) arising independently from a preexisting B-CLL was achieved. Consequently, the patient received local radiotherapy. In our opinion, the designation “Richter's Syndrome”, readily applied in the literature, is inappropriate for this tumor constellation.
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  • 143
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    Archives of gynecology and obstetrics 244 (1988), S. 47-52 
    ISSN: 1432-0711
    Keywords: CA19-9 ; CA125 ; CEA ; Endometrial carcinoma ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Samples of 40 endometrial carcinomas were examined by immunohistochemical methods for CA19-9, CA125 and CEA. CA19-9 was detected in 93%, CA125 in 65% and CEA in 58%. CA19-9 was detected in more than 50% of tumor cells in 14 cases and the same was true for CA125 in six cases. In no tumor was CEA found in more than half the cells. The distribution of CA125 and CEA was markedly more heterogenous than that of CA19-9. There was no statistically significant correlation between immunohistochemical markers on the one hand, and estrogen and progesterone receptor content on the other. A correlation between histological grading and marker detection was only found for CA19-9. CA19-9 was detected in almost all endometrial carcinoma samples, and was the most homogenously distributed. This makes CA19-9 a possibly useful tumor marker for endometrial carcinoma.
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  • 144
    ISSN: 1432-2307
    Keywords: Estrogen ; Progesterone receptor ; Immunohistochemistry ; Prostate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Estrogen (ER) and Progesterone receptors (PR) were demonstrated immunohistochemically on frozen sections from 11 prostatectomy and 7 cystoprostatectomy specimens in the nuclei of various cell types. The periglandular fibrocytes and smooth muscle cells were extensively positive, the interglandular stromal cells were only partly so. Normal basal cells stained focally positive, hyperplastic basal cells stained extensively. The glandular secretory epithelium and atrophic glands were negative. The same findings were obtained in hyperplastic nodules. Both ER and PR also occurred in the urothelium of central prostatic ducts and of the prostatic urethra. The fibrous stroma around the ejaculatory ducts and seminal vesicles was extensively positive while the epithelium was negative. The smooth musculature of the seminal vesicles was only partly positive. On large field sections, the ER as well as the PR were numerically equally distributed throughout the inner zone of the prostate and the prostate proper. 12 prostatic carcinomas (G I–G III) were ER- and PR-negative. Estrogens may contribute to nodular hyperplasia by triggering a stromal proliferation with a secondary inductive epithelial growth. Obviously they do not act directly on prostatic carcinoma but inhibit growth via the hypophyseal-testicualr axis. The biological significance of the PR in the prostate is unknown.
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  • 145
    ISSN: 1432-2307
    Keywords: Human epidermal growth factor ; Foetal submandibular gland ; Pleomorphic adenoma ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The phenotypic expression of the human epidermal growth factor (EGF) was investigated immunohistochemically in human foetal submandibular glands from the 5th to 10th month of gestation, adult normal submandibular glands and 48 cases of pleomorphic adenomas. In foetal submandibular glands, both the terminal buds and primary ducts at the intermediate stage of gestation were positive for EGF, and in particular, the outer layer cells of primary ducts showed strong EGF-immunoreactivity. EGF-positive cells decreased as the gestational stage advanced and only ductal cells were weakly positive for EGF at the terminal stage of gestation. In the adult normal submandibular gland, weak immunoreactivity for EGF was restricted to ductal cells. However, 41 (86%) of the 48 pleomorphic adenomas had EGF-positive cells which were distributed among the ductal, chondroid and myxoid portion. No EGF-immunoreactivity was detected in the solid portion of pleomorphic adenomas. These results suggest that EGF may play an important role in the growth and differentiation of foetal cells as well as the proliferation of tumour cells in pleomorphic adenomas.
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  • 146
    ISSN: 1432-2307
    Keywords: Pituitary Gland ; Amyloid ; Amyloid P Component ; Immunohistochemistry ; Aging
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary To identify amyloid deposits in the anterior pituitary gland, we have immunohistochemical, histochemical and alkaline Congo red staining. The anti-human P component reacted positively with these amyloid deposits, while antisera against prealbumin, AA type amyloid fibril protein and various anterior pituitary hormones were negative. A combination of Congo red and anti-human P component staining was most sensitive and reliable for detection of amyloid in the anterior pituitary glands of 300 randomly autopsied patients. Amyloid deposits increased in parallel with the age of the patients, however, they appeared earlier and more frequently than heretofore reported. Deposition of amyloid was seen initially in the 3rd decade and the positivity rate of amyloid deposits was 73% in the 5th decade. The histochemical characteristics of these pituitary amyloid deposits differed from those of cerebral and systemic deposits, particularly those found in the amyloid of senile systemic amyloidosis.
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  • 147
    ISSN: 1432-2307
    Keywords: Breast ; Carcinoma ; Actin ; Type IV collagen ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Distributions of actin and type IV collagen were investigated immunohistochemically as markers for myoepithelial cells and basement membranes. Carnoy's and Methacarn-fixed, paraffin-embedded tissues from 103 human breast lesions from 103 patients were examined; 65 with carcinomas, 27 with mastopathies, 9 with fibroadenomas and 2 with phyllodes tumours. Fifty-five samples of the normal mammary gland tissue adjacent to tumours were also included for comparison. In normal breast and benign breast diseases, type IV collagen was identified around the mammary glandular cells and actin-positive cells were demonstrated to attach to basement membranes. In noninvasive carcinomas, type IV collagen was found as a continuous lining around a cell nest, while actin-positive cells were usually absent in ductal but quite numerous in lobular carcinomas. In invasive carcinomas, type IV collagen was fragmented or absent and actin-positive cells were very uncommon around the fragmentary basement membranes. These results suggest that the different distributions of myoepithelial cells and basement membrane material is useful in the differential diagnosis of surgical pathology of the breast.
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  • 148
    ISSN: 1432-2307
    Keywords: Collagenous colitis ; Myofibroblasts ; Myoid cells ; Immunohistochemistry ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary We examined 129 colonic biopsies from 21 patients with collagenous colitis, most of whom presented with diarrhoea. Morphometric measurements gave a mean thickness of the subepithelial collagen deposit of 19.5 µ ± 5.1. The trapped fusiform and/or stellate cells within the deposits were identified immunohistochemically as myoid cells, being positive with antibody against smooth muscle cell alpha-actin. Ultrastructurally, these cells have all the characteristic features of myofibroblasts. Similar cells are also present along the crypts, where they were formerly referred to as pericryptal fibroblasts. Although there is still much debate as to the pathogenesis of this condition, we would like to suggest that collagenous colitis is a disease of pericryptal myofibroblasts. During their migration and maturation into the subepithelial region they may synthesize an excess of collagen, under some yet unknown or undefined stimulus/stimuli.
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  • 149
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    Virchows Archiv 413 (1988), S. 341-348 
    ISSN: 1432-2307
    Keywords: Adenomatoid odontogenic tumour ; Keratin ; Vimentin ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Immunohistochemical observations of intermediate sized proteins in five cases of adenomatoid odontogenic tumour (AOT) are described. The immunohistochemical detections of keratins were made with polyclonal antiserum (TK, 41–65 kDa) and three monoclonal keratin antibodies (KL1: 55–57 kDa; PKK1: 40, 45, and 52.5 kDa and nos. 19, 18, 8; K8.12: nos. 16, 13) and vimentin and desmin monoclonal antibodies. Histologically, the tumour epithelia could be divided into two types: type A cells were a spindle or columnar shape and formed solid, ductal, tubular or whorled structures. Type B cells were small and compact cells at the periphery of the A cell-containing focus. Immunohistochemically, the type A cells showed very slight reaction with all antibodies to keratins, whereas the type B cells indicated slight-to-moderate expression of keratin and vimentin, and showed coexpression. Both types of cell showed a negative reaction for desmin. Only one case was associated with cystic lesions, and the cyst-lining was composed of thin squamous epithelium. Keratin expression in this epithelium was strong. In the histogenesis of AOT it was postulated that the tumour cells may have originated from undifferentiated odontogenic epithelium or stratum intermedium cells.
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  • 150
    ISSN: 1432-2307
    Keywords: Lipid rich carcinoma ; Breast ; Immunohistochemistry ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Five cases of lipid-rich carcinomas of the breast were investigated ultrastructurally and immunohistochemically for alpha-lactalbumin (ALA), lactoferrin (Lfr) and human milk fat globule membrane antigen (HMFG-2). Staining for ALA and Lfr showed intensive reaction on nearly all of the tumour cells whereas immunoreaction for HMFG-2 revealed positivity in single cells. All tumours were negative for steroid receptor content. Ultrastructurally the tumour cells showed numerous intracytoplasmic non-membrane bound lipid droplets which were often found within autophagocytic vacuoles. Neither rough endoplasmic reticulum nor Golgi complexes showed any sign of lipid synthesis. Extrusion of lipid droplets and extracellular lipid deposition was not observed. In conclusion, our findings do not justify the consideration of lipid-rich carcinoma of the breast as a clearly defined group of tumours with specific secretory activity. Therefore, the term lipid-rich carcinoma should be used in preference to lipid-secreting, unless there is evidence of active lipid secretion.
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  • 151
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    Virchows Archiv 413 (1988), S. 387-398 
    ISSN: 1432-2307
    Keywords: Islet cell tumours ; Immunohistochemistry ; Neuroendocrine markers ; Human chorionic gonadotropin, alpha subunit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary A comparative immunocytochemical investigation was performed on a series of 59 pancreatic endocrine tumours using a panel of seven markers for neuroendocrine neoplasms: neurone specific enolase (NSE), PGP 9.5, chromogranin A (CgA), PHE5, prealbumin (Pa), HISL-19, and alpha-subunit of human chorionic gonadotropin (α-HCG). Most markers can be separated into two groups characterized by an identical immunoreactive cellular compartment and substantial overlapping in the immunohistochemical results. The first group comprises soluble cytoplasmic proteins such as NSE and PGP 9.5 and is characterized by a diffuse, homogeneous staining of the cell cytoplasm that is not related to the type of hormone produced or the degree of cell differentiation. The second group includes antigens located in the cell secretory granules such as CgA, PHE5, Pa and HISL-19 and is characterized by a heterogenous, often polarized cell staining. The latter markers strongly react with benign glucagonomas and PP-omas and, in contrast with those of the former group, are stricly neuroendocrine-specific. However, they often are less effective in staining insulinomas and malignant tumours. An additional, distinctive and useful characteristic of the HISL-19 antibody was its ability to label the Golgi complex also in tumours with absent granular staining. Finally, α-HCG was found in 9 of 16 malignant tumours (mostly glucagonomas and insulinomas) and in 4 of 43 benign neoplasms (all insulinomas). The latter finding is not in accordance with the reputed specificity of the α-HCG expression by pancreatic endocrine tumours as a marker for tumour malignancy.
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  • 152
    ISSN: 1432-2307
    Keywords: Endothelial cells ; Macrophage ; Lung ; Immunohistochemistry ; Monoclonal antibodies
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The existence of a subpopulation of human vascular endothelial cells (EC) has been demonstrated in the liver with the aid of immunohistochemical techniques. In this study, we investigated the antigenic and functional properties of the vascular EC in human lung. Alveolar capillary EC shared antigens with a peripheral blood monocyte/ macrophage subset capable of presenting soluble antigens and triggering autologous mixed lymphocyte reactions. That is to say that the alveolar capillary EC were HLA-DR+, OKM1−, and OKM5+. In addition, these EC frequently expressed interleukin-1. These facts suggest that alveolar capillary EC may play an important role in immunological responses in the lung. The antigens were, however, absent or only faintly visible on the vascular EC of medium and small vessels. In contrast, Factor VIII/von Willebrand factor antigen (FVIIIRAg), which is produced in vascular EC was heavily stained in the EC of medium and small vessels, but only weakly stained in the alveolar capillary EC. These immunohistochemical findings suggest that in different anatomical compartments in the lung vascular EC express phenotypic properties heterogeneously. They may play differing biological roles or serve different immunological functions in normal and pathological states in the lung.
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  • 153
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    Virchows Archiv 412 (1988), S. 499-513 
    ISSN: 1432-2307
    Keywords: Alveolar soft part sarcoma ; Cytology ; Electron microscopy ; Immunohistochemistry ; DNA analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The type, differentiation and histogenesis of the tumor cells of alveolar soft part sarcoma (ASPS) have been analyzed in a series of ten cases by a light-microscopic, ultrastructural, immunohistochemical and cytologic investigation and quantitative DNA analysis. Four tumors deviated from ordinary ASPS: three were wholly or partly of the so-called pleomorphic variant of ASPS and a fourth tumor showed calcifications of the psammoma body type. The ultrastructural findings and immunohistochemical demonstration of desmin supported the hypothesis of a rhabdomyomatous differentiation and gave no support to epithelial (negative immunoreactions for cytokeratins, epithelial membrane antigen, HMFG-1 and -2, tissue polypeptide antigen (TPA)) or neuroectodermal (negative for S-100 protein, glial fibrillary acidic protein, neurofilaments) differentiation. The negative immunoreactions for vimentin and myoglobin and the positive reaction for neuron specific enolase (NSE) do not exclude a rhabdomyomatous differentiation since in rhabdomyosarcomas the undifferentiated rhabdomyoblasts generally contain vimentin and the differentiated tumor cells contain myoglobin and rhabdomyosarcoma has previously been reported as being positive for NSE. The production of external lamina material peripherally in the tumor cell nests and around vessels in the vascular septa was demonstrated both ultrastructurally and by immunohistochemistry using antibodies against collagen IV and laminin. The cytologic appearance in smears obtained by fine-needle aspiration from a case of the pleomorphic variant showed some resemblance to that of a carcinoma. The seven tumors with an ordinary cell appearance were found to show a diploid DNA-distribution at a quantitative analysis performed on paraffin sections, while the three tumors wholly or partly of the pleomorphic type showed an additional tetraploid peak.
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  • 154
    ISSN: 1432-2307
    Keywords: Breast carcinoma ; Adenocarcinoma ; Tumour cell heterogeneity ; Monoclonal antibody b-12 ; Tumour marker ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary A mouse monoclonal antibody, MAb b-12, has been described previously (Stähli et al. 1985) which reacts with a Mr 350 kD glycoprotein with mucin-like characteristics (Stähli et al. 1987) expressed in cytoplasm and on the surface of human breast carcinoma cell lines (MCF-7 and ZR-75-1). In the present report the immunohistochemical reactivity of this MAb with normal and malignant human tissues is analyzed. Pre-experiments showed that the epitope b-12 is resistant to formalin treatment allowing the use of tissue processed by standard paraffin embedding methods. 167 normal and 408 neoplastic tissues were tested by indirect immunofluorescence or the avidin-biotin complex method. MAb b-12 stained the apical cytoplasm of secretory epithelia and their secretions including the acinar and ductular epithelia of the breast. It reacted with all breast carcinomas independent of their histological type or stage, frequently with all but in some cases with a fraction of the tumour cells. Some other carcinomas, primarily those of adenomatous differentiation, were also reactive. In these, however, the fraction of positive tumour cells was usually lower. The b-12 epitope is thus a marker for normal and neoplastic epithelia with secretory functions, particularly for breast carcinomas of all histological types and stages, and perhaps a differentiation marker for abortive adenomatous differentiation in solid carcinomas of the gastro-intestinal, uro-genital or respiratory tract.
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  • 155
    ISSN: 1432-2307
    Keywords: Immunohistochemistry ; Parathyroid hormone ; Chromogranin A ; Secondary hyperparathyroidism ; Haemodialysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Thirty-seven nodular hyperplastic parathyroid glands obtained by subtotal parathyroidectomy from 11 haemodialysed patients with secondary hyperparathyroidism were examined both pathologically and immunohistochemically. Four consecutive sections of the largest section-surface of each gland were subject to 4 different stains (haematoxyline-eosin, Grimelius, and the immunohistochemical stains for parathyroid hormone and chromogranin A) for comparison of each nodule. It was found that the major part of each nodule consisted of a single cell type with a single pattern of cells. These reacted uniformly to each stain. The mechanism involved in the storage and secretion of the secretory granule appeared to be regulated at the nodule and not at the cell level. The results suggest that the nodules may come from a monoclonal proliferation of a single parathyroid cell. Our present light microscopic immunohistochemical study, failed to demonstrate completely identical immunoreactive positivity of each nodule or each parathyroid cell to PTH. Chromogranin A or secretory protein-I did not indicate the coexistence of PTH and SP-I in the same secretory granule, which was in good agreement with the electron microscopic immunocytochemical study of Arps using bovine parathyroid glands. Our present study, however, provides good evidence that chromogranin A positivity is demonstrable in the human parathyroid gland outside the adrenal medulla and sympathetic nerves.
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  • 156
    ISSN: 1432-2307
    Keywords: Intermediate filaments ; Immunohistochemistry ; Salivary gland neoplasms
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The expression and distribution of cytokeratins and vimentin in fifteen malignant salivary neoplasms were examined by immunocytochemical techniques using, five monoclonal antibodies (mAbs) against different epitopes of Cytokeratins (CKs) (mAbs PKK1, PKK2, and PKK3, identifying CKs 8, 18 and 19, CKs 7, 17 and 19, and CK 18, respectively) and Vimentin (mAbs V9 and V24). Antibody PKK1 gave strong reactions in all neoplasms showing the similarity of these tumours to other digestive system adenocarcinomas. Three general staining patterns of the neoplasms were recognized with respect to the reactivity of mAbs PKK2, PKK3, and V9. Mucoepidermoid cancer, salivary duct carcinoma and a clear cell carcinoma had a higher relative content of CKs 7, 17 and 19 than of CK 18. Adenoid cystic carcinoma showed the same CK pattern but in the periphery of the tumour cords vimentin was readily detected. In two acinic cell carcinomas, the relative content of CK 18 was higher than that of CKs 7, 17 and 19. Furthermore vimentin was expressed in the tumour cells. However, one mucoepidermoid carcinoma showed vimentin expression and two acinic cell carcinomas were vimentin negative and more reative for PKK2 than PKK3. Pecularities in CK expression were seen: squamous areas of mucoepidermoid carcinomas were stained by mAb PKK3 although CK 18 is not present in normal squamous epithelia or in squamous cell carcinomas of tongue and skin. In conclusion, the different salivary neoplasms can be distinguished on basis of IFP content. Such a differentiation fits with current theories of histogenesis, i.e. vimentin is seen in tumours presumed to arise from intercalated duct reserve cells, whilst the vimentin negative neoplasms would be expected to arise in excretory duct reserve cells.
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  • 157
    ISSN: 1432-2307
    Keywords: Gallbladder cancer ; Pyloric gland metaplasia ; Pepsinogen ; Immunohistochemistry ; Paradoxical concanavalin A staining
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Gastric phenotypic expression indicated by paradoxical concanavalin A (Con A) staining for class III mucins and the immunoperoxidase method for pepsinogen (Pg) I and Pg II was found in pyloric gland metaplasia of gallbladder epithelium. Using the same methods, the features of gallbladder cancers and their relationship to pyloric gland metaplasia in the human gallbladder epithelium were studied. Histologically, 57 gallbladder cancers were classified into 5 papillary adenocarcinomas, 29 tubular adenocarcinomas, 8 poorly differentiated adenocarcinomas, 6 signet-ring cell carcinomas, 4 mucinous adenocarcinomas, and 5 squamous cell carcinomas. In papillary and tubular adenocarcinomas, Pg I and/or Pg II staining was detected in 80% and 75.9% of cancers, respectively. Pg II staining was significantly more frequent than Pg I staining. One signetring cell carcinoma also had Pg II activity. Pyloric gland metaplasias all contained class III mucins and were further classified into complete type and incomplete type on the basis of presence or absence Pg I and/ or Pg II activities. A few cancer cells with class III mucins were negative for Pg staining; conversely, a few cells with Pg I and/or Pg II had no class III mucins. Phenotypic diversity in both class III mucin reactivity and Pg activities was observed in gallbladder cancer cells with the pyloric gland cell type. By comparison, pyloric gland metaplasia varied only in Pg activities. A few Pg-positive cancers were found in the gallbladder with Pg-negative pyloric gland metaplasia. The present results clearly indicate the appearance of gastric phenotypic expression in both gallbladder epithelium and gallbladder cancers and suggest the independent induction of pyloric gland metaplasia and cancer with gastric phenotypic expression.
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  • 158
    ISSN: 1432-2307
    Keywords: Cytokeratins ; Middle ear mucosa ; Aural cholesteatoma ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Cytokeratin expression was studied in human middle ear cholesteatoma lesions, using a variety of immunohistological techniques and a wide range of polyclonal antisera and monoclonal antibodies against cytokeratin (CK) subgroups or individual CK polypeptides. The expression of the other cytoskeletal proteins, vimentin and desmin, was also investigated. Middle ear mucosa and epidermal tissues were used as reference tissues. Our investigations also included epithelial structures present in the cholesteatoma perimatrix and in dermal tissues. The results indicate that, compared with epidermal tissues, the expression profile of CKs in cholesteatoma matrix is representative of a hyperproliferative disease. Evaluating the presence of a marker of terminal keratinization - the 56.5 kD acidic CK no 10 - we found supportive evidence of a pronounced retardation of its expression, which did not parallel histological differentiation. In epidermal tissues, the first prickle cell layers are CK10 positive whereas in many cholesteatomas this finding was observed near the stratum granulosum only. Probing the early stages of keratinization -the 58 kD basic CK no 5 and the 50 kD acidic CK no 14 - we regularly observed an extended staining area in the cholesteatoma matrix. In epidermal reference tissues, only the basal and nearest suprabasal layers were convincingly labeled. As a rule, non-epidermal CKs did not belong to the cholesteatoma CK set. However, exceptions to that rule were noticed as a focal or more extended expression of one or more non-epidermal CKs in about half of the cases. Together with the extended CK5 topography, this is further evidence that CK expression is seriously affected by the diseased state. CK expression in the perimatrix is limited to mucous glands, either normal, atrophic or hyperplastic. CKs no 4, 5, 7, 14, 18 and 19, also displayed by middle ear mucosa, were consistently observed. Where ductal arrangements were present, CK10 was also detected, in analogy with the CK10 registration in ductal portions of mucous glands in the external ear canal skin. The absence of CK8 in mucous glands of the perimatrix, however, strongly differentiates these structures from the mucous gland acini and ducti in the external ear canal, where CK8 is systematically expressed. Vimentin staining was restricted to dendritic cells of the matrix (Langerhans cells) and to perimatrix fibroblasts, blood cells and vascular endothelium. Coexpression of CK and vimentin was not observed.
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  • 159
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    Virchows Archiv 414 (1988), S. 69-76 
    ISSN: 1432-2307
    Keywords: Epithelioid cells ; Granuloma ; Immunohistochemistry ; Macrophages ; Spleen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary In six patients with systemic symptoms, four of which had lymphoplasmacytic infiltrates in the bone marrow splenectomy was performed because of suspected malignant lymphoma, with resolution of clinical symptoms. The spleen showed epithelioid granulomas, and neither splenic tissue nor clinical follow-up revealed evidence of malignant lymphoma. Immunohistochemical analysis documented an identical phenotype in epithelioid and giant cells as well as in large numbers of CD4+ lymphocytes and S100+ interdigitating reticulum cells within the granulomas. These cases are interpreted to represent epithelioid granulomatosis with primary and/or predominant manifestation in the spleen.
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  • 160
    ISSN: 1432-2307
    Keywords: Chronic neutrophilic leukemia ; Lymph node ; Immunohistochemistry ; Immature neutrophil
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary An immunohistochemical study was performed on autopsy material from a patient with chronic neutrophilic leukaemia (CNL) using antibodies against various cell lineage-related antigens. Proliferation of immature neutrophils with occasional clusters of erythroblasts and megakaryocytes were noted in the retroperiotoneal lymph nodes, spleen, and kidneys as well as in the bone marrow. Predominance of immature neutrophils in the lymph nodes suggested the emergence of a blast crisis, although there was no increase of blasts in the peripheral blood. Since immature myeloid cells are difficult to distinguish from malignant lymphoid cells on tissue sections, we suggest that immunohistochemical identification of cell lineage-related molecules on these cells is necessary for the more accurate interpretation of lymph node lesions in myeloid neoplasms.
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  • 161
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    Virchows Archiv 412 (1988), S. 247-253 
    ISSN: 1432-2307
    Keywords: Pancreatic osteoclast-type giant cell tumour ; Differentiation ; Histogenesis ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Two cases of osteoclast-type giant cell tumour of the pancreas (OGTP) are presented and compared with similar tumours of other locations and pancreatic carcinomas. One of the tumours was analyzed by immunohistochemical methods. The mononuclear stromal cells and osteoclast-like giant cells, which characterize this very rare neoplasm, reacted with an antibody against vimentin, but were not decorated by antibodies against lysozyme, alpha-1-ACHT, alpha-1-AT. Pleomorphic mononuclear cells in osteoid additionally contained osteonectin and could thus be identified as osteoblasts. Only the tumour glands stained positively with panepithelial keratin antibodies and antibodies against the keratin polypeptides 7, 18, 19. These results demonstrate for the first time the mesenchymal differentiation of the OGTP, which in some cases is also able to form epithelial structures. The immunohistochemical reactions and the characteristic morphology of the tumour show the OGTP to be an entity which must be differentiated from pancreatic carcinoma, especially from its giant cellular subtype.
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  • 162
    ISSN: 1432-2307
    Keywords: Malignant melanoma ; Anti-human-melanoma monoclonal antibody ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The distribution of a recently produced melanoma specific antibody (HMB-45) has been evaluated histochemically on paraffin sections in a large panel of melanocytic and non melanocytic tumours. Results have been compared with the presence of S-100 protein. HMB-45 was shown to be a highly specific antibody being present only in melanomas, junctional melanocytes and histogenetically related neoplasms such as melanocytic neuroectodermal tumour of infancy and, at low levels, on a proportion of peripheral nerve sheath tumours. The high specificity of HMB-45 antibody, coupled with the greater sensitivity of S-100, makes the combined use of these markers practical in the differential diagnosis of skin tumours and of metastatic lesions of uncertain primary site.
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  • 163
    ISSN: 1432-2307
    Keywords: Chromogranin ; Pheochromocytoma ; Medullary thyroid carcinoma ; Carcinoid tumour ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Neuroendocrine cells have histologically common features represented by argyrophilic cytoplasm containing neuroendocrine granules. Neuroendocrine granules are composed of various kinds of peptide hormones, amines, carrier proteins and ATP. Although various kinds of peptide hormones have been detected in neuroendocrine tumours, a peptide hormone has not been required as a standard marker for these tumours. Chromogranin is a purified protein which binds catecholamines specifically and is recognized as a carrier protein. We carried out an immunohistochemical study of chromogranin immunoreactivity in 100 neuroendocrine tumours including pheochromocytomas, carotid body tumours, medullary thyroid carcinomas and carcinoid tumours. Marked immunoreactivity was observed in 85% of carcinoid tumours and 100% of the other tumour types. A non-functioning paraganglioma and a malignant carcinoid tumour without any other detectable marker also showed strong immunoreactivity to chromogranin. Chromogranin immunoreactivity is a useful tool for neuroendocrine tumours.
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  • 164
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    Virchows Archiv 413 (1988), S. 61-68 
    ISSN: 1432-2307
    Keywords: Pituitary adenoma ; Immunohistochemistry ; Follicle stimulating hormone (FSH) ; α-subunit ; Thyroid stimulating hormone (TSH)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Ten FSH producing pituitary adenomas were studied immunohistochemically. 9 cases were in males, and 7 showed elevated serum FSH levels. Immunohistochemically, all cases showed the presence of α-subunit and FSH-β subunits in many tumour cells. These two subunits were frequently colocalized in the same cells. However, the expression of LH-β subunit was extremely low (1 of 10 cases exhibiting occasional LH-β positive tumour cells), although it has been reported that FSH-β and LH-β subunits are colocalized in the same cells of the normal adult pituitary gland. Immunoelectron microscopically, α-subunits and FSH-β were present in the secretory granules and suggested the co-release of subunits or secretion of combined form of FSH. In 7 cases, TSH-β was positive, and in some cases, TSH-β was colocalized in the same tumour cells which contained α-subunit and FSH-β subunit. A few cases also demonstrated immunoreactivity for PRL and ACTH. Our immunohistochemical studies suggest that FSH adenomas are multihormonal and that there is abnormal gene expression in FSH cells with loss of LH-β appearance and co-expression of TSH-β.
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  • 165
    ISSN: 1432-2307
    Keywords: Estrogen receptor ; Gastric carcinoma ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Estrogen receptors (ER) in human gastric carcinomas were examined immunohistochemically using a specific monoclonal antibody to human ER. ER-immunoreactivity (ER-IR) was positive in 30 (27.8%) of the 108 gastric carcinomas examined. ER-IR was located in the nucleus of cancer cells. The incidence of ER-IR positive gastric carcinoma was not significantly different between male and female cases. However, the positive tumour cells were observed in 28 (39.4%) out of the 71 poorly differentiated adenocarcinoma, the incidence being significantly higher than that in well differentiated adenocarcinoma (p〈0.01). There was no significant difference in the incidence of ER-IR between scirrhous carcinoma and non-scirrhous poorly differentiated adenocarcinoma. Synchronous expression of ER and epidermal growth factor receptor was found in 8 of the 26 scirrhous carcinomas (30.8%). Patients with ER-IR positive scirrhous gastric carcinomas showed a much worse prognosis than those with ER-IR negative scirrhous carcinomas.
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  • 166
    ISSN: 1432-2307
    Keywords: Neuroendocrine differentiation ; Immunohistochemistry ; Electron microscopy ; Small cell undifferentiated bladder carcinoma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Small cell carcinoma with the histological appearance of pulmonary small cell carcinoma is a rare tumour in the urinary bladder. In previous case reports the neuroendocrine nature of small cell bladder carcinoma has been accepted, but on review the evidence for true neuroendocrine differentiation appears unsatisfactory. In this study the histological, immunohistochemical and ultrastructural characteristics of three cases of small cell carcinoma of the urinary bladder are described. Ultrastructurally, the cytoplasm of all three tumours contained neurosecretory-type granules and each of the tumours demonstrated positive immunoreaction for two or more neuroendocrine markers, from a panel including neuron-specific enolase, chromogranin A, Leu-7, bombesin and synaptophysin. Although the combination of ultrastructural and immunohistochemical examination obviously offers the strongest evidence in establishing neuroendocrine differentiation, it is argued that immunohistochemistry alone may also yield important information in demonstrating a neuroendocrine nature, provided that at least neuron-specific enolase and synaptophysin are included as markers. The clinical relevance of identifying neuroendocrine differentiation in small cell bladder carcinoma is suggested by the favourable response to combination chemotherapy in two of our cases.
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  • 167
    ISSN: 1432-0568
    Keywords: Atrial specific granule ; Atrial natriuretic polypeptide ; Impulse conducting system ; Immunohistochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The present study was performed to clarify the distribution of ANP-containing cells in the adult rat heart by immunostaining for ANP using antiserum against α-human ANP. ANP-immunoreactive cells were generally present in the atrial walls except for the sinoatrial node. In the ventricular walls, they were distributed in the impulse conducting system, particularly the left bundle branch, Purkinje fibers on the left side of the interventricular septum, and those in the false tendons in the left ventricle, while they were sporadically seen in the atrioventricular node and bundle of His. The immunoreactive cells contained specific granules that were positive for ANP. These findings demonstrate that ANP-containing cells are present in the atrial and ventricular walls.
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  • 168
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    Acta neuropathologica 76 (1988), S. 522-527 
    ISSN: 1432-0533
    Keywords: Esthesioneuroblastoma ; Tyrosine hydroxylase (TH) ; Immunohistochemistry ; Ultrastructure ; Catecholamine-producing tumor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary An esthesioneuroblastoma in a 16-year-old male was studied ultrastructurally and immunohistochemically, using antiserum against tyrosine hydroxylase (TH), a rate-limiting enzyme in the catecholamine-synthesizing pathway. Tumor cells were fairly uniform in appearance, showing scantly cosinophilic cytoplasm and round to oval hyperchromatic nuclei, and were arranged in nests and cords of various sizes. Ultrastructurally, individual tumor cells had well-developed cell organelles including polyribosomes, microtubules, intermediate filaments, centrioles, Golgi apparatus and mitochondria. Secretory-like granules were occasionally found, predominantly in the cell processes. Immunohistochemically, many tumor cells were shown to be immunoreactive for TH. This finding strongly suggested that the present tumor was capable of producing catecholamines and that it might be derived from certain sympathetic neuronal cell nests in the superior nasal cavity.
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  • 169
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    Acta neuropathologica 76 (1988), S. 458-464 
    ISSN: 1432-0533
    Keywords: Immunohistochemistry ; Folliculo-stellate cells ; Pituitary adenomas ; Intermediate filaments
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Folliculo-stellate (FS) cells were studied in 102 human pituitary adenomas by immunohistochemical techniques using antibodies to S-100 protein and intermediate filaments protein. In most pituitary adenomas there were few S-100-positive cells, in contrast, numerous FS cells were found in four of the 54 cases of non-functioning adenomas. Among glial fibrillary acidic protein (GFAP), keratin and vimentin, FS cells showed greatest affinity to vimentin. Stains for desmin or neurofilaments were always negative. Counterstains with GFAP and keratin could demonstrate a small number of double-labelled cells, but mainly disclosed two types of FS cells positive for either GFAP or keratin. Accordingly, FS cells were grossly subdivided into two types: GFAP-positive cells which might be neuroectodermal or glial in origin and keratin-positive cells which might be oral ectodermal or derive from the Rathke's pouch.
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  • 170
    ISSN: 1432-0533
    Keywords: Germ cell tumor ; Suprasellar ; Canine ; Comparative neuropathology ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Suprasellar germ cell tumors were identified in five young adult to middle-aged dogs. Clinical signs included depression, mydriasis, ptosis and visual deficit. At necropsy large extramedullary masses were found on the ventral surface of the rostral brain stem. Histologically four were characterized by sheets and nests of moderately pleomorphic round cells resembling seminoma, admixed with larger vacuolated cells, glandular formations with secretory material, and occasional foci of squamous differentiation. The fifth case was more homogenous, with germinomatous elements predominating. Immunohistochemical examination of three tumors revealed positive staining for alpha fetoprotein. Although only one case showed intracranial metastasis, these tumors were considered malignant due to the marked local invasion and destruction. The veterinary literature is reviewed, and we propose that two tumors previously reported as craniopharyngioma be reclassified as germ cell tumors. One other tumor previously reported as an unclassified suprasellar tumor was identified, which also demonstrated features of a germ cell tumor.
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  • 171
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    Acta neuropathologica 75 (1988), S. 557-565 
    ISSN: 1432-0533
    Keywords: Hypertension ; Blood-brain barrier ; Immunohistochemistry ; Light microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary A transient increase in blood pressure was induced in 15 male Sprague Dawley rats by clamping the upper abdominal aorta for 8–10 min. Three rats served as controls. The brains were fixed by perfusion 2 h or 7 days later. Evan's blue-albumin (EBA) was used for macroscopic evaluation of the blood-brain barrier (BBB) integrity. Extravasated plasma albumin, fibrinogen and fibronectin were demonstrated by immunohistochemistry on paraffin sections. Glial fibrillary acidic protein (GFAP) was visualized in the same way. Parallel sections were analyzed for possible parenchymal changes associated with the BBB breakdown. Multiple focal areas of BBB opening were seen in the brains of the three rats killed 2 h after the hypertensive episode. The plasma proteins were present in the vascular wall, extracellular space and within certain neurons. Shrunken acid fuchsin positive neurons were seen in some areas of extravasation. After 7 days, in 5 out of 12 rats a few local lesions with EBA leakage and positive immunostaining for plasma proteins were seen. Structurally these lesions were characterized by shrinkage, fuchsinophilia and disintegration of neurons and proliferation of astrocytes. Thus, a transient opening of the BBB by acute hypertension may lead to permanent tissue damage.
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  • 172
    ISSN: 1432-0533
    Keywords: Prostaglandin F2-alpha ; Immunohistochemistry ; Transient increase ; Hippocampus ; Purkinje cell
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The changes in prostaglandin F2-alpha (PG F2α) staining over 3 days of recirculation in both fore-and hindbrains were studied. Five minutes of global ischemia was produced in 24 rats by Pulsinelli's method with hypotension around 50 mm Hg of mean arterial blood pressure. Eight rats (including three pretreated with indomethacin) were recirculated for 5 min, three for 1 h, five for 2 h and five for 3 days. Five normal rats without occlusion of vessels served as controls. The brains were snap frozen. Ten-micrometer cryosections were stained for PG F2α by the indirect immunofluorescence method after fixation in carbodiimide and in Zamboni's solution. Positive staining for PG F2α was noted in pial vessels in all normal and ischemic rats. Recirculated rats revealed the strongest reaction at 5 min after recirculation in blood vessels and in neuronal cytoplasm (especially in hippocampi and in Purkinje cells). The intensity of staining was markedly reduced after 1 h. Rats pretreated with indomethacin showed less increase in staining. The above results indicate that recirculation after ischemia produces a transient increase in PG F2α in blood vessels and neurons of both fore- and hindbrains.
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  • 173
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    Acta neuropathologica 76 (1988), S. 128-134 
    ISSN: 1432-0533
    Keywords: Early human fetal brain ; Immunohistochemistry ; Vimentin ; Neurofilament protein (NFP) ; Glial fibrillary acidic protein (GFAP)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary To assess the cytogenesis of the central nervous system we studied the spinal cord and the cerebrum in 11 human embryos and fetuses of gestation age 7–25 weeks immunohistochemically using anti-vimentin, anti-neurofilament protein (NFP), anti-neuron-specific enolase (NSE), anti-glial fibrillary acidic protein (GFAP), anti-S-100 protein, anti-Leu 7 and anti-myelin basic protein (MBP) antibodies. Vimentin was demonstrated in ventricular cells at 7 weeks and older. NFP-68-kDa and-160-kDa components were observed in neuroblastic cells of the neural tube at 7 weeks. NFP (68 and 160 kDa) was mainly located in the marginal zone of the spinal cord and the cerebrum at 8–9 weeks. NSE was not found in the neural tube at 7 weeks, although NSE was demonstrable at 9 weeks both in the spinal cord and in the cerebrum. GFAP-positive cells started to appear at 9 weeks in the spinal cord and at 15 weeks in the cerebrum, respectively. S-100 immunoreactivity was almost coincident with GFAP. S-100, however, was observed in more numerous glioblastic cells. Leu 7 was detected at 7 weeks and located in the neuropil of the central nervous tissue. MBP was not demonstrable in this study. Our study indicates that neuronal differentiation occurs much earlier than glial differentiation in the human brain and that neuronal and glial cell classes do not coexist in the ventricular zone of the early human fetal brain.
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  • 174
    ISSN: 1432-0533
    Keywords: Glial fibrillary acidic (GFA) protein ; S-100 protein ; Immunohistochemistry ; Hirschsprung's disease
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The supportive cells of the enteric nervous system were examined in gut tissues from 15 patients with Hirschsprung's disease by means of immunohistochemistry, utilizing antisera to glial fibrillary acidic (GFA) protein and S-100 protein. In the normoganglionic segment, GFA protein immunoreactivity was predominantly found in association with the myenteric plexus and to a lesser extent in the submucous plexus. On the other hand, the extrinsic, hypertrophic nerve fasciculi were selectively immunostained with GFA protein antiserum throughout the entire length of the aganglionic intestinal walls from all children studied. The large fasciculi were numerous in the distal aganglionic segment and commonly appeared in the intermuscular zone and submucosal connective tissue. Both small-and mediumsized nerve fasciculi with GFA protein immunoreactivity were also encountered within the circular muscle layer of the proximal aganglionic segment. A subpopulation of supportive cells within the hypertrophic nerve fasciculi showed immunoreactivity for GFA protein, while all supportive elements of these fasciculi were stained for S-100 protein. The intrinsic nerve fibers within the circular muscle layer of normoganglionic segments were stained for S-100 protein, but not for GFA protein. The present study supports our previous findings that two types of supportive cells can be differentiated by immunohistochemistry in the enteric nervous system, utilizing antisera to GFA protein and S-100 protein. It is also concluded that the demonstration of GFA protein by immunohistochemical methods favors the diagnosis of aganglionic colons with Hirschsprung's disease, since GFA protein immunoreactivity is confined to the extrinsic, hypertrophic nerve fasciculi characteristic of aganglionic bowels.
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  • 175
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    Acta neuropathologica 76 (1988), S. 245-252 
    ISSN: 1432-0533
    Keywords: Pleomorphic xanthoastrocytoma ; Fibrous xanthoma of the meninges ; Fibrous histocytoma ; Glial fibrillary acidic protein ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary A case of a fibrous xanthomatous tumor of the meninges is reported. This is a rare tumor of childhood in which the characteristic pleomorphic histology contrasts with the good clinical prognosis [26]. These tumors were reclassified as pleomorphic xanthoastrocytomas (PXA) due to their glial fibrillary acidic protein (GFAP) positivity [27]. In the present tumor, GFAP was absent from nearly all cell bodies in most of the leptomeningeal regions of the tumor but could be detected with greater frequency at the cortical-leptomeningeal border zones and in the areas in which the tumor had infiltrated the cortex. All the tumor cells expressed vimentin and in, addition, most expressed α-1-antitrypsin, α-1-antichymotrypsin, tartrate-resistant acid phosphatase, common leukocyte antigen, and OKM1. This spectrum of marker staining corresponded not only to the pattern observed in two cutaneous fibrous histiocytomas and one malignant fibrous histiocytoma, but also to the results previously published in the literature with regard to fibrohistiocytic tumors. By contrast, this spectrum of monocytic-histiocytic marker staining was not seen in gliomas. We, therefore, regard the PXA as a mesenchymal tumor of the meninges, identical to benign fibrous histiocytomas elsewhere in the body. The possible reasons why this mesenchymal tumor can show GFAP positivity in the leptomeningeal border zone are discussed.
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  • 176
    ISSN: 1432-0568
    Keywords: Atrial natriuretic peptide ; Fetal heart ; Spontaneously hypertensive rat ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The development of immunoreactive atrial natriuretic peptide (ANP) was studied in fetal hearts of spontaneously hypertensive (SHR) and compared to normotensive Wistar-Kyoto (WKY) rats. While SHR fetal hearts were noticeably less developed than those of WKY at 10 and 11 days gestation, both strains showed ANP immunoreactive cells in some but not all primitive heart tubes. At 12 days additional ANP immunoreactive cells appeared in formative trabeculae of the ventricle and atrium. ANP cells were also observed in the myogenic layer of the truncus and bulbus arteriosus and their derivatives from 11 through 16 days, but not at 18 days. In both strains, there were more ANP cells in the left ventricle than in right beginning at day 13. There were no obvious strain differences in the developmental pattern and timing of ANP producing cells. However, on the day of birth, staining was reduced in hearts from some WKY newborn pups compared with hearts from SHR newborns and ventricular staining was reduced in both strains when compared to fetal hearts. These observations indicate that ANP is one of the earliest peptide hormones produced and that the predisposition to genetic hypertension does not appear to influence the development of ANP.
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  • 177
    ISSN: 1432-0568
    Keywords: Pancreatic islet ; Insulin ; Glucagon ; Ontogeny ; Immunohistochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Ontogenetic appearances of glucagon, insulin and tyrosine hydroxylase (TH) were immunohistochemically investigated on developing pancreatic islets of rats. Glucagon immunoreactivity appeared first in some epithelial cells (g-cells) of the dorsal anlage of the pancreas on day 11.5 of gestation. On day 12.5, g-cells increased in number manufacturing the primitive islets, in which some cells appeared to be immunoreactive for insulin (i-cells) and about 40% of g-cells indicated also a slight immunoreactivity for insulin (g/i-cells). Afterwards, all the islet cells, especially g-cells, increased in number, and almost half of g-cells were g/i-cells. After day 16.5 of gestation, numerical increase of the cells with insulin immunoreactivity exceeded that of the cells with glucagon immunoreactivity, and about one fifth of g-cells were g/i-cells. After 20.5 days, however, no g/i cells were found. On day 16.5 of gestation, the immunoreactivity for TH appeared in occasional cells of the islets, but the cells did not show immunoreactivity for glucagon or insulin. It is concluded that the progenitor cells of the pancreatic islets appear to synthesize both glucagon and insulin by day 20.5 of gestation, but differentiate giving rise to mature A and B cells of adult isoets afterward.
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  • 178
    ISSN: 1432-0568
    Keywords: Salivary gland ; Differentiation ; Immunohistochemistry ; Cytokeratins ; Vimentin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Adult and developing salivary glands were investigated using five monoclonal antibodies against cytokeratins (CKs) and vimentin. Acinar cells displayed mainly CK 18 whereas CKs 7, 17 and 19 were only detected in duct and myoepithelial cells. All epithelial and myoepithelial cells were unreactive for one vimentin antibody (Vim 9) whereas with the other (Vim 24), myoepithelial cells and basal cells of excretory ducts were stained. Fetal cells showed the CK pattern of duct cells. At gestational week 18, a reaction for both vimentin antibodies could be found in basal cells of terminal tubules. Although vim 9 reactivity has been shown for a number of salivary neoplasms, it has not been detected in any adult epithelial salivary tissue. The finding of this reactivity in the fetal gland indicates that the expression of this intermediate filament protein in certain salivary neoplasms may be a sign of dedifferentiation resulting in the expression of a filament pattern found in an earlier stage of gland development.
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  • 179
    ISSN: 1432-0568
    Keywords: Fish ; Muscle development ; Myosin ; Immunohistochemistry ; Histochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary InDicentrarchus labrax (the sea bass) the differentiation of lateral muscle fibres occurs at different stages and in different ways in the superficial (red), intermediate (pink) and deep (white) regions of the myotome. At hatching the myotomes are composed of presumptive white and red fibres, the latter forming a superficial monolayer present only near the transverse septum. At this stage, differences between the fibre types are mainly ultrastructural. From their different reactions with isoform-specific antibodies to mullet myosin, and the appearance of histochemical mATPase activity, it appears that in both red and white muscle fibres there is a transition in myosin composition from an early larval form (L1R and L1W respectively) to a late larval form (L2R and L2W) and then to the isoforms typical of adult red and white muscle. The transition from L1W to L2W in the deep muscle occurs very rapidly and early in larval life (between 10 and 28 days), whereas the equivalent transition in the superficial muscle (from L1R to L2R) is a gradual process beginning in fibres near the trasverse septum and spreading hypo- and epi-axially as this layer grows around the deep muscle. The definitive adult forms (AR and AW), distinguishable by the appearance of characteristic histochemical myosin ATPase activity, are present in the superficial red muscle by 80 days, but later in the deep white muscle (by 20 months), respectively. Compared to the superficial red and deep white muscle, the intermediate (pink) muscle layer first appears relatively late (80 days), but then acquires the histo- and immunohistochemical profile characteristic of the adult form much more rapidly. The mosaic appearance of the deep white muscle is first seen during larval life. However, at this stage the smaller fibres of the mosaic have a different histo- and immunohistochemical profile from that seen in the adult small white fibres, which may indicate different mechanisms of histogenesis of new fibres in the deep muscle layer in the early and adult stages.
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  • 180
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    Anatomy and embryology 178 (1988), S. 297-307 
    ISSN: 1432-0568
    Keywords: Rabbits ; Islands of Langerhans ; Electron microscopy ; Immunohistochemistry ; Regulation of islands
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary In addition to “external” signals conveyed by the circulation or the nervous system, the pancreatic islets obviously are regulated also by “internal” (intra-islet) signals, e.g. by the islet hormones: insulin (B-), glucagon (A-), and somatostatin (D-) cells are able to affect the secretion of the heterologous cell types. It is, however, unclear whether this functional cooperation between islet cells occurs by an intercellular route (paracrinia sensu strictore), by intraislet “portal” vessels, or by the systemic circulation. These likely interactions are limited by islet anatomy. To identify the anatomical basis for the mutual functional relationships between the islet cells, islets of Langerhans in the rabbit pancreas were completely analyzed in immunostained serial semithin (0.5 μm) sections. The islets were found to be largely heterogenous. They were classified in three basic types: a) polycellular islets, composed of all established endocrine cells, and including two subtypes of islets, b) bicellular islets, containing only B- and A-cells or B- and D-cells, and c) monocellular islets, exclusively made up of B-cells. Concerning the modes of paracrine regulation of islet cells, the findings suggest primarely an endocrinous route of transport of the islet peptides to heterologous endocrine cells. The corresponding functional cooperation between islet cells probably is mediated rather by the systemic circulation than by intra-islet portal vessels.
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  • 181
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    Anatomy and embryology 178 (1988), S. 327-336 
    ISSN: 1432-0568
    Keywords: Pituitary gland ; Immunohistochemistry ; Three-dimensional reconstruction ; Rathke's pouch
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary In order to clarify the environmental factors which are involved in the development of the primordium of the pituitary gland such as cell-cell interactions, a three-dimensional reconstruction of this organ and its surrounding tissues was carried out. Pituitary material was obtained from human fetuses mainly during the period of organogenesis. Rathke's diverticulum was found to stretch rostrally from the stomodeal epithelium to the middle of the mesoderm, and already by the 5th week of fetal growth, it was clearly seen to be involved with the diencephalon. The area of contact between Rathke's pouch and the diencephalon gradually moved from the rostral to caudal regions and, after 13 weeks of development, had a position similar to that found in the newborn infant. Among the cells forming Rathke's pouch, it was found that the closer their relationship was to the diencephalon, the greater were their epithelial characteristics. When the relationship of such cells to the diencephalon was weaker, their differentiation to endocrine cells occurred earlier. Immunohistochemically, that portion of the pituitary primordium which has a close relationship with the diencephalon, later to become the pars intermedia, showed an adrenocorticotropic hormone (ACTH) immunoreactivity later than that of the pars anterior. On the other hand, in the 21st fetal week, nearly all of the cells of the pars intermedia were found to be ACTH-positive. This finding is thought to indicate a close connection between the physical contact between the brai (diencephalon) and the pituitary primordium and the development of the pars intermedia; the differentiation of ACTH cells. The surface of the epithelium of Rathke's cavity continues to increase at least until the 21st fetal week, so the growth of the epithelium of Rathke's pouch is thought to be heavily involved in the growth of the primordium of the pituitary gland in the early stages of development.
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  • 182
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    Anatomy and embryology 178 (1988), S. 429-440 
    ISSN: 1432-0568
    Keywords: Ontogeny ; Sheep brain ; Serotonin ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Using immunohistochemistry with specific antiserotonin anti-sera, the ontogeny of serotonergic neurons was studied in the foetal sheep brain. Serotonergic-immunoreactive perikarya first appeared rostrally on day 25 of pregnancy, in the medio-ventral part of the mesencephalic flexure, and caudally, on day 28, in the medio-ventral part of the cervical flexure. The development of this system is very rapid, because on day 40 of gestation, all serotonergic nuclei present in the adult were visible. Compared with other species such as rodents or primates, serotonin appears early in the sheep nervous system, and the development of the serotonergic system is even more rapid. Serotonergic immunoreactivity was seen in some cell bodies in the growing adenohypophysis between days 40 and 50. This phenomenon has not been observed in other species. Because serotonin appears very early and is present in growing areas of the nervous system, it could play a trophic role in the development and maturation of the sheep central nervous system, as has been described previously in other species.
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  • 183
    ISSN: 1432-0568
    Keywords: Immunohistochemistry ; Choline acetyltransferase ; Olfactory bulb ; Modified glomerular complex ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The main and accessory olfactory bulbs (MOB and AOB) of the rat were immunohistochemically stained with a monoclonal antibody against choline acetyltransferase (ChAT) in order to know the difference in the distribution patterns of cholinergic fibers between these two structures. A few ChAT-immunoreactive cell bodies were found in the superficial and middle parts of the external plexiform layer (EPL) of the MOB, in the granule cell layer (GCL) of the MOB, and in the GCL of the AOB. The frequency in appearance of these cells was 0.9 cells/section in the MOB and 0.3 cells/section in the AOB. While the glomerular layer (GL) and the superficial part of the EPL were most densely innervated in the MOB, the internal plexiform layer received the richest innervation in the AOB. There were no immunoreactive structures in the olfactory nerve layer of the MOB and in the vomeronasal nerve layer and glomerular layer of the AOB. In addition to a relatively homogenous distribution of cholinergic fibers in the MOB and AOB, there were several foci of very dense network of immunoreactive fibers at the posterior level of the OB. These foci formed a part of the modified glomerular complex that was recently identified using 2-deoxyglucose method and was presumed to be related to suckling behaviour in the neonatal rat.
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  • 184
    ISSN: 1432-0568
    Keywords: 5-Hydroxytryptamine ; Immunohistochemistry ; Motor neuron pool ; Lateral motor column ; Posture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The pattern of distribution of serotonin positive fibers in the motor nuclei of the chick spinal cord was examined immunohistochemically by using an antiserum against serotonin. A dense aggregation of serotoninergic fibers was located around anterior horn cells in the cervical spinal cord. In the brachial spinal cord, serotoninergic fibers were densely aggregated in the medial motor column and in the parts of the lateral motor column. There were two regions of serotonin immunoreactivity in the lateral motor column of the brachial spinal cord; one located in the ventromedial regions where a dense aggregation of serotoninergic fibers was found, and the reminder of the lateral motor column where only a few serotoninergic fibers were observed. The region containing a dense cluster of serotoninergic fibres around profiles of motoneuron somata and proximal dendrites appears to correspond to motor neuron pools of flexor muscles. In the thoracic spinal cord a high density of serotoninergic fibers was found in the motor nucleus. In the lumbosacral spinal cord (segments LS1–LS8) serotoninergic fibers were not observed in the medial motor column. However, there were five regions in the lateral motor column, where a high density of serotoninergic fibers was found. These very likely correspond to motor neuron pools of muscles which extend the hip joint.
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  • 185
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    Archives of dermatological research 280 (1988), S. 93-96 
    ISSN: 1432-069X
    Keywords: α2-Macroglobulin ; Monoclonal antibodies ; Immunohistochemistry ; ELISA ; Blister fluid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Monoclonal antibodies recognizing the antiproteolytic compound α2-macroglobulin (MG) were used for immunohistological studies on normal human skin. MG-specific immunoreactivity was found to be localized to the papillary dermis and to be concentrated in the region of the epidermodermal junction. In view of these findings and the possible involvement of proteolytic enzymes and their inhibitors in blister formation, we asked whether MG occurs in the fluid of experimentally induced blisters. MG was identified (by western-blotting) and quantified (by a monoclonal antibody based enzyme immunoassay) in the fluid of experimentally induced suction blisters. Taken together, MG is present in such blister fluid in concentrations 6 times lower than in serum, but still in an antiproteolytic range. These findings allow suggestion of a possible role for the antiproteolytic compound MG in blister formation.
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  • 186
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    Acta neuropathologica 76 (1988), S. 564-573 
    ISSN: 1432-0533
    Keywords: Polyradiculoneuritis ; Lymphocytic infiltrates ; Immunohistochemistry ; Vascular endothelium ; Blood-nerve barrier
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Human sural nerve biopsies of eight cases with acute, subacute and chronic polyradiculoneuritis were studied by means of immunohistochemistry to characterize the inflammatory infiltrates. In addition, the structural changes of the endoneurial blood vessels were examined by electron microscopy, since both factors are likely to contribute to disturbances of the blood-nerve barrier. By use of six monoclonal antibodies, it was shown that the inflammatory infiltrates in cases with more acute polyradiculoneuritis are predominantly recruited by Leu 3a- and Leu 4-positive T lymphocytes. In more chronic polyradiculoneuritis beside of few Leu 3a-positive and Leu 4-positive T lymphocytes also B cells occurred. Leu M3-positive macrophages were detected in all cases with floride myelin degeneration. Since immunoreactivity for antigens of the HLA-D-locus (Leu-HAL-DR and Leu 10) were present on the infiltrating mononuclear cells, it can be postulated that they represent active and immunocompetent cells. Ultrastructurally, the amount of pinocytotic vesicles in the endothelial cells of the endoneurial blood vessels was increased. Moreover, a prominent folding of the luminal and abluminal surface of vascular endothelial cells and diminution of the intercellular tight junctions were observed. These findings appear suitable to explain the increased leakage of serum proteins across the blood-nerve barrier in polyradiculoneuritis sharing general features of cell-mediated immunity.
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  • 187
    ISSN: 1432-0533
    Keywords: Blood-brain barrier ; Vasogenic brain edema ; Plasma proteins ; Immunohistochemistry ; Hyperosmolar solutions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary An immunohistochemical study was carried out on rat brain to determine if a transient opening of the blood-brain barrier (BBB), leading to extravasation of serum albumin, is also associated with exsudation and cellular uptake of fibronectin and fibrinogen. Both of them might exert important biological effects provided that they pass the BBB and come into contact with cells of the brain parenchyma. Hyperosmolar solutions of urea or mannitol were infused in the carotid artery for 30 s to open the BBB and the animals were killed at various time intervals thereafter. Formaldehyde-fixed, paraffin-embedded material was used for immunohistochemical demonstration of extravasated proteins by an avidin-biotin peroxidase technique. Multifocal, often confluent areas of widely different sizes with signs of albumin extravasation were observed both in the grey and the white matter of the cerebral hemispheres exposed to the hyperosmolar solutions. Much less pronounced changes were observed in rats given an intracarotid saline infusion alone. Immunoreactive material indicating extravasation of fibronectin and fibrinogen was present in the infused cerebral hemispheres but albumin immunoreactivity was much more widespread. Reaction product was observed in vascular walls, presumably in extracellular spaces and in nerve cells. Immunoreactivity in the perikaryon of neurons formed different patterns in various cells. Agranular type most probably represents accumulation of the proteins in lysosomal organelles after pinocytotic uptake into the neuron. The second so-calleddiffuse variety is presumably the result of a severe nerve cell injury with an uncontrolled leakage of proteins into the cytoplasm. Our results indicate that vascular walls, extracellular spaces, glial cells and neurons will be exposed to extravasated fibronectin and fibrinogen as well as to albumin and that antigenic sites in such compounds remain for a long period after the BBB opening. In addition, there are indications that carotid infusions of hyperosmolar solutions may cause nerve cell injuries in regions with BBB opening. These findings have obvious clinical and experimental significance.
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  • 188
    ISSN: 1432-0533
    Keywords: Neoplastic angioendotheliosis ; Malignant lymphoma ; B cell lymphoma ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Frozen cerebral and renal tissue sections of an autopsied “neoplastic angioendotheliosis (NAE)” case were investigated immunohistochemically using monoclonal and heterologous antibodies to lymphocyte, monocyte, endothelial, epithelial and histiocytic antigens. In both tissues, positive stainings for surface immunoglobulin (sIg) μ and ϰ, but not λ, were observed in most of the neoplastic cells. These cells were also positive for other B cell markers (BA-1, Leu-12 and HLA-DR). No distinct staining was observed in the neoplastic cells with antibodies to T lymphocyte (OKT-11 and Leu-1) or monocyte (OKM-1) markers. Posive stainings were observed only in some small round lymphoid cells which were distributed sporadically in and around blood vessels and were considered to be reactive. No positive staining was observed in the neoplastic cells with antibodies to endothelial (factor VIII), epithelial (cytokeratin) or histocytic (lysozyme) antigens. Thus, our NAE case was shown to be of monoclonal B cell lymphoma in nature.
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  • 189
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    Acta neuropathologica 75 (1988), S. 337-344 
    ISSN: 1432-0533
    Keywords: Experimental allergic encephalomyelitis ; Serum thymic factor ; Suppressor T cell ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Acute experimental allergic encephalomyelitis (EAE) was induced in Hartley guinea pigs and Lewis rats, which were then treated with synthetic serum thymic factor (FTS). When a dose of 30 μg/100 g body weight of FTS was subcutaneously administered to the animals on days — 1 (before inoculation), 4, 9 and 15 intermittently, clinical symptoms of acute EAE were suppressed. Histopathological evaluation showed that the severity of EAE in FTS-treated guinea pigs was less than in unteated guinea pigs. Immunohistochemical examination showed that the numbers of OX6+, W3/25+, W3/13+ and OX19+ cells in FTS-treated rats were less than in untreated rats and that the number of OX8+ cells in FTS-treated rats was greater than in untreated rats. These findings suggest that FTS induced OX8+ cells in inflammatory lesions and suppressed inflammation in acute EAE.
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  • 190
    ISSN: 1432-0533
    Keywords: Gaucher disease ; Brain ; Immunohistochemistry ; Macrophages ; Astrocytes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Splenectomy in children with the Norrbottnian type of Gaucher disease is followed by increased blood levels of glucosylceramide and imparied neurological and mental status. High blood levels are associated with an increased accumulation of glucosylceramide in perivascular Gaucher cells in the brain compared to non-splenectomised cases. Surrounding the Gaucher cell infiltrates there is loss of neurons and slight demyelinaton in the brain parenchyma. The brains of four cases with the Norrbottnian type of Gaucher disease were examined by immunohistochemical stains in an attempt to further characterize the perivascular Gaucher cells and to examine the reactions of the vessel walls and brain parenchyma to the accumulation of Gaucher cells. The perivascular storage cells showed granular staining with antibodies to muramidase and α1-antichymotrypsin confirming that they are blood-derived macrophages belonging to the monocyte-macrophage system. The Gaucher cells contained material positive for antisera to plasma proteins strongly suggesting that large molecules (including glucosylceramide) can escape from the blood and be taken up by the macrophages in Gaucher disease. The storage cells were surrounded by a reticulin network stained by antisera to collagen type III, type IV and laminin. The infiltrates were bounded from the brain parenchyma by a membrane strongly positive with antiserum for the basal lamina protein collagen type IV and laminin. The formation of a basal lamina around the Gaucher cell cuffs probably constitutes a protective phenomenon governing the brain parenchyma against the foreign cells. A focal loss of neurons but only minor loss of axons could be demonstrated with the antiserum to neurofilament. The brain parenchyma surrounding the Gaucher cell infiltrates showed marked astrogliosis in the anti-glial fibrillary acidic protein stain. In the two cases previously shown to have higher blood levels of glucosylceramide there were astrocytes positive for plasma proteins indicating passage of plasma proteins into the brain, this was not seen in the non-splenectomised cases. The additive effect of low-grade tissue damage in the vicinity of the Gaucher cell infiltrates is probably enough to explain the increased neurological symptoms and mental retardation following splenectomy in the Norrbottnian type of Gaucher disease.
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  • 191
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    Acta neuropathologica 75 (1988), S. 535-537 
    ISSN: 1432-0533
    Keywords: Astrocytosis ; Hyperplasia ; Quantitative analysis ; Immunohistochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The number of astrocytes in an atrophic human brain was counted with the aid of immunohistochemistry for glial fibrillary acidic protein. Microscopically, astrocytosis was remarkable in the cerebral neocortex and white matter. Taking the cortical atrophy into consideration, however, the total number of astrocytes in the cortical layers II – VI was not increased. The number of astrocytes in the white matter was not increased either. It is indicated that astrocytosis does not always mean hyperplasia (net increase of total number) of astrocytes.
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  • 192
    ISSN: 1432-0533
    Keywords: Serotonin ; Mouse ; Neostriatum ; Seizure ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Changes occurring in serotonin neurons during hyperthermia-induced convulsions were examined by means of a modified immunohistochemical method. All mice (8–12 weeks of age) exposed to the temperature of 50°C had convulsions, showing a generalized tonic and/or clonic pattern. Immediately after the convulsions, the animals were perfused transcardially with a fixative. A significant reduction in serotonin immunoreactivity was observed in the neostratum (caudate-putamen complex) of the mice which had hyperthermia-induced seizures, while the serotonin immunoreactivity remained unchanged in the neocortex and paleostriatum. These results suggest that serotonin may be an important mediator in the mechanism of hyperthermia-induced convulsions or that the susceptibility of serotonin neurons to a convulsive state is greatest in the neostriatum.
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  • 193
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    Acta neuropathologica 77 (1988), S. 33-38 
    ISSN: 1432-0533
    Keywords: Rathke's cleft cyst ; Immunohistochemistry ; Pituitary hormones ; Intermediate filament ; Squamous metaplasia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary An immunohistochemical study was made of ten cases of asymptomatic and three cases of symptomatic Rathke's cleft cyst. The cysts in the asymptomatic cases had monolayer columnar or cuboidal epithelium. Within the epithelium, cells which were positive for at least one of the pituitary hormones were found. The rate of positive reactions for these six pituitary hormones was between 70% and 100%. In contrast, the cysts in the symptomatic cases had an epithelium which was partly stratified squamous and partly squamous epithelium, and none of the pituitary hormones were found in them. Therefore, when a Rathke's cleft cyst enlarges to the extent that clinical symptoms are produced, we consider that changes have already occurred in structure and function of the cyst epithelium. In addition, we believe there is a tendency for monolayer epithelia to undergo squamous metaplasia and for cells which are positive for pituitary hormones to change into non-granulated cells.
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  • 194
    ISSN: 1432-0533
    Keywords: K virus ; Murine papovavirus ; Central nervous system ; Endothelial cells ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The murine papovavirus K causes fatal pneumonia in infant mice, but an asymptomatic infection in older mice. In order to establish whether the virus affects the central nervous system in the course of systemic infection, we carried out morphological and immunohistochemical studies on the experimentally infected mice. BALB/c mice, less than 4 days of age, were inoculated with K virus either intraperitoneally or intracerebrally. When the animals were moribund, usually 10 days or so, after inoculation, their brains were removed and examined. Acutely infected mice showed only minor changes: intranuclear eosinophilic inclusions in very rare capillary endothelial cells of the brain. However, immunoperoxidase studies, using specific antibody to K virus, revealed that a number of brain cells had positive nuclear staining. These nuclei were distributed throughout the brain, without an apparent site of predilection. Double-immunostaining showed that virtually all cells whose nuclei were positive for viral antigen were endothelial, because their cytoplasm was positive for factor-VIII or vimentin. There were no nuclei positive for viral antigen in astrocytes, as determined by positive staining for glial fibrillary acidic protein or glutamine synthetase. By electron microscopy, clusters of K virus particles were found only in the nuclei of brain capillary endothelial cells. Although these endothelial cells showed degeneration of varying degree, their basement membranes remained relatively intact and there was no disorganization in the endfeet of contiguous astrocytes. Neurons and glial cells had normal ultrastructures. Therefore, this study has demonstrated that there is involvement of central nervous system during systemic K virus infection and that the infection involves predominantly brain capillary endothelial cells.
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  • 195
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    Journal of cancer research and clinical oncology 114 (1988), S. 291-296 
    ISSN: 1432-1335
    Keywords: Leu-M1 antigen ; Immunohistochemistry ; Thyroid carcinoma ; Medullary carcinoma ; Prognosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Leu-M1 antigen is a monocyte/granulocyte-related marker known to be consistently expressed in the Reed-Sternberg cells of patients with Hodgkin's disease and to be present in tumour cells of a variety of non-haematopoietic neoplasms, most of them adenocarcinomas. The biological significance of this aberrant reaction has not yet been clarified. Recently, however, we have demonstrated that marked epithelial Leu-M1 immunoreactivity significantly correlated with an unfavourable clinical course in papillary carcinomas of the thyroid gland. The findings of the present study obtained from surgical speciments of 39 tumours suggest that Leu-M1 immunostaining also provides significant prognostic information in patients with medullary carcinoma (MC) of this organ. Irrespective of other morphological and clinical features, local recurrences occurred 2.9 times (P〈0.005) and death resulting from tumour occurred 4.3 times (P〈0.03) more frequently among MCs with marked Leu-M1 positivity (〉15% tumour cells positively stained) in comparison to tumours with only slight or absent immunoreactivity. A significantly higher recurrence rate of intense Leu-M1-positive MCs was even evident when comparing only tumours of stage pT1-3N0M0 (P〈0.005). Our findings infer that Leu-M1 immunostaining might be of clinical relevance to the selection of different aggressive adjuvant therapeutic procedures to be used in MCs with high or low malignant potential.
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  • 196
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    Experimental brain research 71 (1988), S. 388-398 
    ISSN: 1432-1106
    Keywords: γ-Aminobutyric acid (GABA) ; Glutamic acid decarboxylase (GAD) ; Somatostatin ; Coexistence ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The distribution of somatostatin-like immunoreactive (SS-LI) material and its colocalization with glutamic acid decarboxylase (GAD)-like immunoreactivity were studied in the rat hippocampus and dentate gyrus neurons using immunohistochemistry. In the dentate gyrus and CA1 region, SS-LI perikarya were concentrated in the hilus and in the stratum oriens, respectively, whereas immunoreactive cell bodies were rarely seen in other layers. Approximately half of the SS-LI neurons of the CA3 region were situated in the stratum oriens, the other half being scattered in strata pyramidale, lucidum and radiatum. About 90% of SS-LI neurons were also GAD-like immunoreactive, whereas about 14% of GAD-like immunoreactive (GAD-LI) neurons were SS-like immunoreactive. The percentage of GAD-LI neurons which were also immunoreactive for SS varied from one layer to the other. This percentage was about 30% in the hilus of the dentate gyrus and in the stratum oriens of the CA1 and CA3 regions; it was 5–10% in the strata pyramidale, lucidum and radiatum of the CA3 region and reached only 2% in the granule cell layer and molecular layer of the dentate gyrus and in the stratum pyramidale and stratum radiatum in the CA1 region. These observations indicate that the majority of SS-LI neurons in the rat hippocampal formation are a subpopulation of GABAergic neurons.
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  • 197
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    Experimental brain research 72 (1988), S. 523-534 
    ISSN: 1432-1106
    Keywords: Neuropeptides ; Tyrosine hydroxylase ; Colocalization ; Coexistence ; Hypothalamus ; Periaqueductal grey matter ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Double-labeling combined with elution-restaining immunofluorescence techniques were used to analyze the extent of coexistence among the peptides cholecystokinin (CCK), peptide histidine-isoleucine (PHI)/vasoactive intestinal polypeptide (VIP), substance P and the catecholamine-synthesizing enzyme tyrosine hydroxylase in neurons of the supramammillary region and mesencephalon of the rat. Approximately 50% of the PHI/VIP-containing perikarya and about 25% of the CCK-positive cell bodies in the supramammillary region exhibited coexistence of both peptides. Only a very minor portion of these double-labeled neurons were also found to contain immunostaining for tyrosine hydroxylase (indicative of dopamine in these cells). A low percentage of the neurons contained the enzyme plus either CCK- or PHI/VIP-like immunoreactivity. A low proportion of the tyrosine hydroxylase-positive neurons in this region contained substance P-like immunoreactivity and vice versa. In other areas, small numbers of neurons in periventricular and periaqueductal regions were found to be immuno-stained for CCK, PHI/VIP and tyrosine hydroxylase. Single examples of triple-labeled (CCK-PHI/VIP-TH) somata were infrequently observed in the ventral tegmental area. These data provide further evidence of peptide/peptide and peptide/monoamine coexistence in the central nervous system. The demonstration of CCK-PHI/VIP colocalization (possibly including a minor dopaminergic component) and of substance P and tyrosine hydroxylase coexistence within neurons of the supramammillary region, which has widespread projections to many areas of the forebrain, suggests that these neuropeptides may coexist in some of these pathways and perhaps be co-released in several different regions of the brain.
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  • 198
    ISSN: 1432-1106
    Keywords: GABA ; ACh ; Coexistence ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The coexistence of immunoreactivities for choline acetyltransferase (ChAT) and glutamic acid decarboxylase (GAD) and/or gamma-aminobutyric acid (GABA) was revealed in some brain regions of the rat, using the peroxidase-antiperoxidase method. Consecutive 40 μm thick vibratome sections were incubated in different antisera and those cells which were bisected by the plane of sectioning so as to be included at the paired surfaces of two adjacent sections were identified. The coexistence of the immunoreactivities for ChAT and GAD or GABA in the same cell could thus be determined by observing the immunoreactivity of the two halves of the cell incubated in two different antisera. In the retina, cerebral cortex, basal forebrain and spinal cord, colocalization of ChAT-like and GAD-like or GABA-like immunoreactivities was observed in some cell types, whereas no such colocalization was observed in cells in the striatum or brainstem. In the retina, the majority of ChAT-like immunoreactive (ChAT-LI) amacrine cells contained GABA-like or GAD-like immunoreactivity. About half of the ChAT-LI neurons in the cerebral cortex showed GABA-like immunoreactivity. In the basal forebrain only a small proportion of ChAT-LI neurons (0.6%) contained GAD-like immunoreactivity. In the spinal cord, about one-third of ChAT-LI central canal cluster cells and about half of ChAT-LI dorsal horn cells showed GAD-like and/or GABA-like immunoreactivities. These observations indicate the possible coexistence of two classical transmitters, GABA and acetylcholine, in various brain regions and spinal cord of the rat.
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  • 199
    ISSN: 1432-1106
    Keywords: Neurotransmitter ; Immunohistochemistry ; 3-D reconstruction ; Confocal microscopy ; Rat ; Lamprey
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary A new method for three-dimensional reconstruction of transmitter-identified neurons is presented which involves “en bloc” immunofluorescence histochemistry and confocal scanning microscopy. The technique was applied to different types of neurons in the rat brain and lamprey spinal cord. Thick sections or tissue “blocs” (50–200 μm thick) were incubated with antisera against neuropeptides or monoaminergic markers, followed by fluorescent secondary antibodies. Three-dimensional reconstructions were obtained by scanning the preparations in sequential focal planes with a thin laser beam, while sampling the emitted light in each focal plane. The method is convenient and can be applied to a wide variety of neuron types. The reconstructions obtained are accurate since the “optical serial sections” of the specimen are perfectly aligned, and optic disturbances such as “halo” phenomena do not occur.
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  • 200
    ISSN: 1432-1106
    Keywords: S-100 protein ; Calcineurin ; Mammalian brain ; Avian brain ; Vertebrate brain ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The cellular and topographic localization of calcineurin and S-100 protein was examined immunohistochemically in the mammalian and avian brain. Calcineurin immunoreactivity in both the avian and mammalian brain was located only in neuronal cells. S-100 protein was localized mainly in the glial and Schwann cells within the mammalian brain. However, in the avian brain, neuronal cells in certain regions such as the paleostriatum primitivum and the cerebellum, as well as other non-neuronal cells, exhibited S-100 protein immunoreactivity. A distinct difference was demonstrated in the macroscopic topographic distribution patterns of S-100 protein immunoreactivity between the mammalian and avian brains, while the patterns of calcineurin distribution were essentially identical. In addition, we provided calcineurin- and S-100 protein-immunocytochemical results for the turtle, frog and fish brain.
    Type of Medium: Electronic Resource
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