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  • Springer  (148)
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  • 1
    ISSN: 1435-604X
    Keywords: Photosensitizer ; Photodynamic therapy ; mTHPC ; Temoporfin ; Pharmacokinetics ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Physics , Technology
    Notes: Abstract A Phase I photodynamic therapy (PDT) clinical trial was carried out with Temoporfin (Foscan®, mTHPC) at the Departments of Otolaryngology at Orebro Medical Center (OMC) and Long Island Jewish Medical Center (LIJMC). A range of drug doses, consisting of 0.3, 0.15, 0.075 and 0.0375 mg kg−1, were utilized. Light treatment was performed on the sixth day after injection of the photosensitizer mTHPC. Photodynamic therapy was done on prostate cancer (six cases), bronchial cancer (one case), nasopharyngeal cancer (three cases), laryngeal cancer (eight cases), mesothelioma (one case), laryngeal papilloma (five cases) and basal cell nevus syndrome (one case). A number of patients were treated more than once. Plasma was collected and analysed at 1, 24, 48, 72, 96, 120 and 144 h and at 2 weeks post-injection, to follow the loading and clearance rate of the photosensitizer. Normal and malignant tissues were collected immediately prior to PDT, chemically extracted, and analysed for drug content spectrofluorometrically. Plasma drug levels were proportional to the dose. The half-life of the drug was 45.4 h across the entire dose range. The ratio of the drug in the tumour compared to normal adjacent mucosa was in the range of 2–3. There were no significant adverse effects. These data establish the basis for full clinical trials.
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  • 2
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    Calcified tissue international 16 (1974), S. 37-44 
    ISSN: 1432-0827
    Keywords: Human ; Dentine ; Glycosaminoglycan ; Chondroitin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract The glycosaminoglycans of root dentine from permanent human teeth were studied. The tissue was digested with papain and the glycosaminoglycans were precipitated with cetylpyridinium chloride and recovered by centrifugation. A small quantity of keratan sulphate was recovered from the supernatant after fractionation on cetylpyridinium chloride and Ecteola-cellulose. The cetylpyridinium chloride-precipitated glycosaminoglycans were studied by infra-red spectrophotometry, cetylpyridinium chloride cellulose chromatography and electrophoresis on cellulose acetate. Individual hexosamines were determined on an autoanalyzer. The results indicate that chondroitin-4-sulphate is the major glycosaminoglycan and that chondroitin-6-sulphate, hyaluronic acid, dermatan sulphate and a nonsulphated galactosaminoglycan are also present in minor quantities.
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  • 3
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    Journal of comparative physiology 185 (1999), S. 297-304 
    ISSN: 1432-1351
    Keywords: Key words Odor coding ; Learning ; Enhanced sensitivity ; Rabbit ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The olfactory system is faced with a particular problem – the high dimensionality and inherent unpredictability of the chemical world. Most natural odorants encountered in everyday life are complex mixtures of many different volatiles. This means that from the outset the olfactory system has to contend with a great and often unpredictable diversity of molecules, making it difficult for stable primary features of the chemical world to be mapped onto the sensory surface. One solution to such unpredictability is provided by learning. Learning confers flexibility, enabling individuals of a given species to acquire and make use of the most appropriate information in a particular environment. Two examples of this are presented: learning of maternal odors in neonatal rabbits, including evidence that the sensory surface itself may be influenced by environmental conditions so as to enhance sensitivity to molecules of particular ecological relevance, and cross-cultural human studies suggesting that experience with everyday odors influences not only the way these are evaluated, but also their perceived intensity. It is concluded that an adequate understanding of odor coding and olfactory function will not be possible without taking such experience-dependent factors into account.
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  • 4
    ISSN: 1420-9071
    Keywords: Human ; bone ; thrombospondin ; growth factors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Thrombospondin (TSP) is a multifunctional glycoprotein which is synthesised by several cell types including osteoblasts, and incorporated into the extracellular matrix (ECM) of these cells. The function and regulation of TSP in bone is not clear. In this study, using a long term culture model of human osteoblast-like cells, we examined the distribution of TSP in the ECM and its modulation by added estradiol. In this model the osteoblast-like cells form a regular multilayer which continues to increase in depth up to 50 days post confluence. In the ECM of these cultures and in 19-week fetal bone, the bone markers osteocalcin and alkaline phosphatase were diffusely distributed in the matrix. In contrast, labelling for TSP was concentrated, confined to the banded collagen and its immediately adjacent ECM. This pattern of labelling resembled that of the growth factors transforming growth factorβ-I (TGFβ), and insulin-like growth factor-I (IGF-I), with which TSP label co-localised. Labelling intensities were comparable between fetal bone and the in vitro material for TSP, TGFβ and IGF-I. TSP label was present by 10 days post confluence, reached a maximum by 20 days, and declined slowly thereafter, a time course which was similar to that of IGF-I. Incubation of osteoblast-like cell cultures with 17β estradiol resulted in an increase in multilayer depth and a maximal 3-fold increase in TSP labeling at 30 days as well as approximately 2-fold increases for TGFβ and IGF-I. The dose-response relationship for these responses to estradiol treatment was biphasic with maximal increases at 10−10 M–10−11 M of added estradiol. Treatment with 17α estradiol produced labelling intensities that were not significantly different from controls. Studies with other cell types have suggested that TSP may be involved in modulation of growth factor activity. The similarities between TSP, TGFβ and IGF-I, in terms of their distribution and regulation by 17β estradiol treatment, may indicate a role for TSP in modulating bone cell proliferation and function through interaction with local growth factors.
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  • 5
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    Calcified tissue international 28 (1979), S. 227-231 
    ISSN: 1432-0827
    Keywords: Enamel ; Human ; Proteins ; Analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Pure human mature enamel was prepared using a careful microdissection technique. After EDTA dissolution, the soluble proteins were recovered representing a concentration of 0.035% in the initial enamel. When the samples were analyzed with polyacrylamide gel electrophoresis, Coomassie Brilliant Blue staining revealed only one sharp fast migrating band, whereas o-toluidine blue, methylene blue, Amido Black 10B, and pyronine red G showed a thin double band at the same migration distance. Ultracentrifugation studies suggested that the proteins were of low molecular weight or of weak density. Absorption spectra showed a strong absorbance at 260 nm. After hydrolysis, amino acid analyses yielded a composition of 25% Gly, 13.5% Glu, 11% Ser, and 11% Pro. Cysteine measured as cysteic acid was present at 2%, and 2% hydroxyproline was found. A carbohydrate content of 15% was estimated by the anthrone method. Glucose, galactose, mannose, and fucose, identified through gas chromatography, were in a molar ratio of 9:4:3:1. Thus the organic matrix of adult human enamel consists of one or possibly two acidic glycoproteins.
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  • 6
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    Calcified tissue international 7 (1971), S. 175-190 
    ISSN: 1432-0827
    Keywords: Bone ; Human ; Age ; Glycosaminoglycans
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les glycosaminoglycanes de l'os humain compact du fémur sont identifiés et leur concentration et distribution sont déterminées à partir de cas d'autopsie, d'âges variables, allant du nouveau-né à des sujets de 94 ans. L'absence d'altérations post-mortem des glycosaminoglycanes osseux est déterminée au niveau de l'os fémoral de chien, au bout de 72 heures. Le chondroitine-4-sulfate constitue le glycosaminoglycane majeur de l'os humain: de plus, une fraction mineure semble être constituée par l'acide hyaluronique. Les proportions de glycosaminoglycanes varient inversement en fonction, de l'âge chez des sujets en pleine croissance: mais aucun changement n'est observé plus tardivement. La distribution des glycosaminoglycanes est identique chez les sujets adultes. Seules de faibles différences, concernant cette distribution, sont relevées. Chez les sujets jeunes elles sont, peut-être, liées à des changements de poids moléculaire peu élevés ou au sulfate chondroitine sulfaté, peu élevé et, chez un sujet vieil à une proportion plus grande d'acide hyaluronique.
    Abstract: Zusammenfassung Die Glykosaminoglykane aus menschlicher Compacta des Oberschenkels wurden charakterisiert und ihre Konzentrationen und Muster an Autopsien von Menschen verschiedenen Alters (Neugeborene bis 94jährige) festgelegt. Fällungen mit Cetylpyridin und Fraktionierungsmethoden wurden angewendet. Am Hundefemur konnte festgestellt werden, daß sich das Glykosaminoglykanmuster im Knochen bis zu 72 Std post mortem nicht veränderte. Als wichtigstes Glykosaminoglykan des menschlichen Knochens wurde das Chondroitin-4-sulfat identifiziert; dazu wurde ein kleiner Anteil versuchsweise als Hyaluronsäure eingestuft. Die Konzentrationen der Glykosaminoglykane standen in umgekehrtem Verhältnis zum Alter des im Wachstum stehenden Menschen, jedoch wurde nach dessen Abschluß keine Veränderung mehr beobachtet. Das Glykosaminoglykanmuster sieht für alle erwachsenen Menschen ähnlich aus. Nur kleine Abweichungen von diesem Muster konnten bei jungen Menschen festgestellt werden; möglicherweise sind sie auf eine leichte Änderung im Molekulargewicht oder auf eine schwache Sulfatierung des Chondroitinsulfates zurückzuführen. Dies gilt ebenso für einen alten Menschen, dessen Knochen einen höheren Anteil an Hyaluronsäure aufweisen.
    Notes: Abstract Glycosaminoglycans from human femoral compact bone were characterized and their concentrations and patterns were determined in autopsy cases of various ages, newborn to 94 years of age. Cetylpyridinium precipitation and fractionation methods were used. Absence of postmortem changes of bone glycosaminoglycan pattern for as long as 72 hours was checked on dog femoral bone. Chondroitin-4-sulphate was identified as the predominating glycosaminoglycan of human bone; in addition a minor part was tentatively identified as hyaluronic acid. Concentrations, of glycosaminoglycans were inversely related to age in growing subjects, but no change was noted later in life. The glycosaminoglycan pattern had a similar appearance for all of the adult subjects. Only small deviations from this pattern had a similar for young subjects, possibly due to low molecular weight change or low sulphated chondroitin sulphate; and for one old subjects to a higher proportion of hyaluronic acid.
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  • 7
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    Calcified tissue international 6 (1970), S. 260-264 
    ISSN: 1432-0827
    Keywords: Bone ; Human ; Tissue ; Histology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé De l'os spongieux humain est utilisé pour déterminer la viabilité de cellules osseuses après exposition, pendant un temps déterminé, à l'air et au sérum physiologique. La réaction de Feulgen, ainsi que la culture de tissus sont utilisées pour déterminer la viabilité des cellules osseuses. Il apparait qu'aucun ostéocyte ne survit après 30 minutes d'exposition à l'air et au sérum physiologique. Les cellules de la moëlle ne survivent pas à 2 heures d'exposition ö l'air, ou 6 heures d'exposition au sérum physiologique. Le nombre de cellules qui survivent après action de l'air et du sérum physiologique diminue progressivement en fonction de l'augmentation du temps d'exposition.
    Abstract: Zusammenfassung Poröser Knochen von Erwachsenen wurde zur Bestimmung der Überlebenszeit von Knochenzellen verwendet, nachdem er während bestimmter Zeiten der Luft oder physiologischer Kochsalzlösung ausgesetzt wurde. Um diese Überlebenszeit zu bestimmen, wurde die Feulgen-Reaktion gleichzeitig mit der Gewebekultur durchgeführt. Daraus läßt sich schließen, daß keine Osteocyten überleben, nachdem sie 30 min der Luft oder der Kochsalzlösung ausgesetzt waren. Markzellen überlebten nicht, wenn sie 2 Std der Luft oder 6 Std der Kochsalzlösung ausgesetzt waren. Die Anzahl überlebender Zellen nimmt ab, wenn die Dauer, während welcher sie der Luft oder der Kochsalzlösung ausgesetzt werden, zunimmt.
    Notes: Abstract Adult human cancellous bone was used to determine the viability of bone cells after timed exposure to air and saline. To determine the viability of the bone cells, the Feulgen reaction was used in conjunction with tissue culture. It is concluded that no osteocytes survived after 30 minutes exposure to air or saline. Marrow cells do not survive after 2 hours exposure to air, or 6 hours exposure to saline. The number of surviving cells after exposure to air or saline progressively diminishes as the time of exposure increases.
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  • 8
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    Immunogenetics 49 (1999), S. 438-445 
    ISSN: 1432-1211
    Keywords: Key words PA28 ; Proteasome ; Gene structure ; Evolution ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Two proteasome activators PA28α and β, which have been implicated in antigen processing for loading class I MHC molecules, are synthesized in response to Ifn-γ. The human genes encoding these activators (PSME1 and PSME2, respectively) were analyzed by sequencing. Each gene comprised 11 exons, consistent with gene duplication during vertebrate evolution. The intron/exon organization of both genes was highly conserved, the major difference being the absence of the exon encoding the lysine and glutamic acid-rich 'KEKE' motif in PA28β. Two other genes of relevance to the immune system were located close to those for PA28 at 14q11.2 including ISGF3G, a protein involved in transcription after IFNα signalling. These sequences were also characterized.
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  • 9
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    Immunogenetics 48 (1998), S. 40-46 
    ISSN: 1432-1211
    Keywords: Key words Immunoglobulin ; Genes ; Kappa light chain ; Human ; Antibody repertoire
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  We describe a hitherto unknown functional IGKV gene, VkLa, belonging to the IGKV1 subgroup with exon 2 having only 94% similarity to the closest known IGKV gene, 1–13/1D-13 (L4/L18a). Genomic DNA sequences spanning from 5’ of the decanucleotide box to 3’ of the heptamer (649 bp) were cloned and sequenced from four individuals. The new gene encodes the conserved amino acids in the exons and contains no apparent defects in known regulatory intron sequences such as pd-box, dc-box, TATA-box, CCCT-elements, splice-sequences, initiation codon, and heptamer sequence. VkLa is therefore potentially functional and, correspondingly, we found transcripts of properly rearranged VkLa with somatical hypermutations. VkLa was found in 12 of 57 (21%) healthy Caucasians by a nested polymerase chain reaction and subsequent sequencing of exon 2. This finding shows that there is more inter-individual variation in the available IGKV gene repertoire than was hitherto assumed. Finally, we describe a minor correction in the IGKV1D-43 (L23) gene sequence.
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  • 10
    ISSN: 1432-1211
    Keywords: Key words Antigen presenting cells ; Dendritic cells ; Cell surface molecule ; Antigen receptor ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Dendritic cells (DC) are specialist antigen presenting cells which capture antigens in the periphery, migrate centrally, and present the processed antigens in the context of major histocompatibility complex and appropriate co-stimulatory molecules to T lymphocytes for the initiation of an immune response. DEC-205 has been identified as a putative antigen-uptake receptor, which is expressed abundantly on mouse DC. The recently cloned mouse DEC-205 cDNA predicts a molecular structure which has a marked similarity to the macrophage mannose receptor. Using reverse transcriptase-polymerase chain reaction (RT-PCR) and cDNA library screening, we obtained the full coding region of human DEC-205 cDNA from the Hodgkin’s disease-derived L428 cell line. The predicted protein structure is a type I transmembrane protein of 1722 amino acids consisting of a signal peptide, cysteine-rich domain, fibronectin type II domain, ten carbohydrate recognition-like domains, transmembrane domain, and a cytoplasmic tail. Human DEC-205 is 77% identical to the mouse protein with completely conserved cysteines. The DEC-205 gene (LY75) was mapped to chromosome band 2q24 by somatic cell hybrid panel analysis and fluorescent in situ hybridization. Northern blot analysis detected 7.8 and 9.5 kilobase DEC-205 transcripts in myeloid, B lymphoid, and Hodgkin’s disease-derived cell lines. RT-PCR analysis indicated that immature blood DC contain a barely detectable amount of DEC-205 transcripts but these were markedly increased upon differentiation/activation.
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  • 11
    ISSN: 1432-1211
    Keywords: Key words NK cells ; Human ; Surface molecule ; Lectin superfamily ; NK gene complex
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Natural killer (NK) cells constitute the third major population of lymphocytes. They possess the inherent capacity to kill various tumor and virally infected cells and mediate the rejection of bone-marrow grafts in lethally irradiated animals. A large family of NK cell receptors belong to the C-type lectin superfamily and are localized to the NK gene complex on Chromosome (Chr) 6 in the mouse and Chr 12 in the human. Genes in the NK gene complex encode type II receptors and examples include the families of NKR-P1, Ly-49, and NKG2 receptors. Examples of other C-type lectin-like NK cell receptors that occur as individual genes are CD94, CD69, and AICL. Here we report the molecular characterization and chromosomal mapping of a human lectin-like transcript (LLT1) expressed on NK, T, and B cells and localized to the NK gene complex within 100 kilobases of CD69. The cDNA encodes a predicted protein of 191 amino acid residues with a transmembrane domain near the N-terminus and an extracellular domain of 132 amino acid residues with similarity to the carbohydrate recognition domain of C-type lectins. The predicted protein of LLT1 shows 59 and 56% similarity to AICL and CD69, respectively. The predicted protein does not contain any intracellular ITIM motifs, suggesting that LLT1 may be involved in mediating activation signals.
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  • 12
    ISSN: 1432-1211
    Keywords: Key words HLA ; Microsatellite loci ; Microsatellite typing ; Human ; MHC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The human genome contains a large number of interspersed microsatellite repeats which exhibit a high degree of polymorphism and are inherited in a Mendelian fashion, making them extremely useful genetic markers. Several microsatellites have been described in the HLA region, but allele nomenclature, a set of broadly distributed controls, and typing methods have not been standardized, which has resulted in discrepant microsatellite data between laboratories. In this report we present a detailed protocol for genotyping microsatellites using a semi-automated fluorescence-based method. Twelve microsatellites within or near the major histocompatibility complex (MHC) were typed in the 10th International Histocompatibility Workshop homozygous typing cell lines (HTCs) and alleles were designated based on size. All loci were sequenced in two HTCs providing some information on the level of complexity of the repeat sequence. A comparison of allele size obtained by genotyping versus that obtained by direct sequencing showed minor discrepancies in some cases, but these were not unexpected given the technical differences in the methodologies. Fluorescence-based typing of microsatellites in the MHC described herein is highly efficient, accurate, and reproducible, and will allow comparison of results between laboratories.
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  • 13
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    Immunogenetics 48 (1998), S. 312-323 
    ISSN: 1432-1211
    Keywords: Key words Recombination ; Gene conversion ; Allele ; MHC ; HLA ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Some alleles of the major histocompatibility complex (MHC) genes have a reticulate pattern of evolution, probably resulting from the exchange of segments by gene conversion or recombination. Here we compare the extent and patterns of reticulate evolution among the classical class I and class II loci of the human MHC using the recently developed compatibility and partition matrix methods. A complex pattern is revealed with substantial differences among loci in the extent and pattern of reticulation. Extremely high levels of reticulation are observed at HLA-B and HLA-DPB1, high levels at HLA-A and HLA-DRB1, moderate levels at HLA-C and HLA-DQB1, and low levels at HLA-DQA1. The reticulate events are concentrated in the exons encoding the highly variable, peptide-binding domains, suggesting that the sequence combinations produced by these events are maintained by natural selection.
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  • 14
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    European journal of clinical pharmacology 55 (1999), S. 349-352 
    ISSN: 1432-1041
    Keywords: Key words Ischaemic stroke ; Vinpocetine ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract Objectives: To determine whether vinpocetine decreases short- and long-term case fatality and proportion of dependent survivors if administered within 2 weeks of stroke onset. Methods: All published and unpublished trials were attempted to be identified using the standard search strategy of the Cochrane Collaboration Stroke Review Group, using MEDLINE searches performed with all known manufacturer code names and trade names of vinpocetine and by contacting manufacturers of vinpocetine to give information of all randomised controlled trials on vinpocetine in stroke. Researchers who participated in trials on vinpocetine in Hungary were asked for further information. Only truly randomised, unconfounded clinical trials that compared the effect of vinpocetine to either placebo or another reference treatment for acute stroke where treatment started no later than 14 days after stroke onset were eligible for inclusion. Data synthesis and analysis was performed using the Cochrane Review Manager software (RevMan version 3.0). Results: Among the identified studies on vinpocetine in stroke, only one fulfilled the selection criteria for inclusion in the review. No death occurred in the study groups and no statistically significant difference was found in dependency between the treatment and the placebo groups. No adverse effects were reported. Conclusions: Based on only one small randomised controlled unconfounded study, presently there is not enough evidence to decide whether the administration of vinpocetine does or does not decrease case fatality and dependency in acute stroke.
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  • 15
    ISSN: 1432-1041
    Keywords: Key words CYP2D6 ; Genetic polymorphism ; Metoprolol ; Pharmacokinetics ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract Aim: The aim of the present paper was to compare the pharmacokinetics of metoprolol in homozygous Caucasian volunteers for the wild-type CYP2D6 allele (CYP2D6*1/CYP2D6*1) and heterozygous (CYP2D6*1/CYP2D6*4) Caucasians. Methods: Thirty-six unrelated healthy male Caucasians were screened for two of the most frequently occurring mutant alleles (CYP2D6*3 and CYP2D6*4) using polymerase chain reaction (PCR). Twenty-four volunteers with a genotype suggesting a rapid hydroxylator phenotype were enrolled in a bioequivalence trial and each received in a randomized, cross-over fashion one of the three formulations compared. Each formulation contained 200 mg metoprolol tartrate/(tablet). In each of the three periods of the trial, one of the formulations was administered under fasting conditions in the morning on 4 consecutive days. Blood for quantification of metoprolol was drawn immediately before the last dose and in selected time intervals thereafter. A sensitive and specific high-performance liquid chromatography (HPLC) method with fluorescence detection was applied for the quantification of metoprolol. Pharmacokinetic parameters were determined for each subject and statistically compared in two groups of 16 homozygous (CYP2D6*1/CYP2D6*1) and six heterozygous (CYP2D6*1/CYP2D6*4) volunteers. Results: Significant differences between homozygous and heterozygous individuals were observed for all pharmacokinetic parameters. The AUC in the course of one those interval of 24 h (AUCτ), minium steady-state concentration (Cmin ss) and average steady-state concentration (Cav ss) values for heterozygous individuals were more than twice those of individuals. Significantly higher values for Css max , t1/2, half-value duration (HVD) and mean residence time (MRT) were also observed in heterozygous volunteers. The higher concentrations of metoprolol in heterozygous individuals also had pharmacodynamic consequences, namely, greater heart rate and blood pressure reduction.
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  • 16
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    Cell & tissue research 121 (1971), S. 358-376 
    ISSN: 1432-0878
    Keywords: Leukocyte ; Peripheral ; Human ; Ultrastructure ; Osmium-Zinc-Impregnation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung Leukozytenkonzentrate des peripheren Blutes von gesunden Versuchs-personen wurden in 2,5%iger Glutaraldehyd-Lösung (0,1 M Na-Cacodylat-Puffer, pH 7,3) vorfixiert und nach der Methode von Maillet (1959) mit OsO4 + ZnJ2 inkubiert. Dickschnitte wurden mit dem Mikroanalysator (Siemens) auf ihren Gehalt an Osmium, Zink und Jod analysiert und die Dünnschnitte mit dem Elektronenmikroskop untersucht. Bei der Mikroanalyse ließen sich Osmium und Zink, nicht aber Jod nachweisen. Die elektronenmikroskopischen Untersuchungen ergaben folgende Resultate: Beträgt der pH-Wert der Inkubationslösung 5,6, so findet sich eine starke Os/Zn-Imprägnation des Golgi-Apparates, des Kernspaltes und des Endoplasmatischen Retikulums bei sämtlichen Arten der weißen Blutzellen. Die Matrix der Mitochondrien dieser Zellen ist im allgemeinen weniger stark imprägniert. Auch in den Lysosomen der Monozyten und Plasmazellen sind geringfügige Os/Zn-Präzipitate nachweisbar. Es findet keine Reaktion in den Leukozytengranula und in den Zentriolen statt. Dagegen zeigen die Lipidtropfen der eosinophilen Granulozyten eine sehr starke, homogene Os/Zn-Imprägnation. Diese ist bereits nach 40 min Inkubation voll ausgeprägt. Bei einer Schnittkontrastierung mit Uranylacetat werden die Os/Zn-Präzipitate wieder herausgelöst, wenn die Kontrastierung länger als 2 min dauert. Eine Bleikontrastierung hat keinen Einfluß auf die Os/Zn-Präzipitate. Bei Erhöhung des pH-Wertes der Inkubationslösung auf 6,2 kommt es zur starken Imprägnation des Externums der eosinophilen Granula, während Kernspalt, Golgi-Apparat und Mitochondrienmatrix keine Reaktion zeigen. Die Imprägnationsdichte der Lipidtropfen der eosinophilen Granulozyten ist pH-unabhängig.
    Notes: Summary “Buffy coats” of normal human peripheral blood were fixed in 2.5% glutaraldehyde (buffered with 0.1 M Na-cacodylate, pH 7.3) and incubated with a mixture of osmium tetroxide and zinc iodide (pH 5.2) after the method of Maillet (1959). Utilizing an electron probe microanalyser (Siemens), the authors demonstrated the presence of osmium and zinc in the specimen, whereas the presence of iodide could not be proved with certainty. The ratio of osmium and zinc in the precipitates ranges from 3/2 to 4/1. Electron microscopic studies of the leukocytes have lead to the following results: the Golgi apparatus, the perinuclear cleft and the endoplasmic reticulum (ER) of all types of leukocytes are strongly impregnated with osmium/zinc, whereas the matrices of the mitochondria are less impregnated. Osmium/zinc precipitates have also been detected in the lysosomes of monocytes and plasma cells. No reaction was evident in the leukocytic granules or centrioles. The presence of lipid droplets in eosinophils has been demonstrated. These droplets show a dense homogeneous osmium/zinc impregnation which reaches its maximum after 40 minutes of incubation, compared with much longer incubation times (5–6 hours) for other cellular components. Setting the pH of the osmium tetroxide-zinc iodide solution to 6.2 resulted in a dense impregnation of the “externum” of the eosinophilic granules. At this same pH, the perinuclear cleft, Golgi apparatus and mitochondrial matrix did not show a definite reaction. The extent of impregnation of the lipid droplets of the eosinophils showed no dependency on pH. Care must be taken when a stain containing uranyl acetate is used since this substance dissolves the Os/Zn precipitates. Exposure of the precipitates to the uranyl acetate for longer than 2 minutes results in gradual dissolution within complete dissolution in 10 minutes.
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  • 17
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    Cell & tissue research 122 (1971), S. 145-159 
    ISSN: 1432-0878
    Keywords: Brain stem ; Human ; Nucleus reticularis lateralis ; Lipofuscin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung Die Gestalt des Nucleus reticularis lateralis, des Nucleus interfascicularis hypoglossi, des Kerns von Roller und einer bisher unbekannten Zellschicht (Repagulum cuneati) wird beschrieben. Dafür werden Serien von 800 μ dicken, mit Aldehydfuchsin gefärbten Schnitten herangezogen. Durch diese Methode werden die in den Nervenzellen enthaltenen Lipofuscinkörnchen elektiv dargestellt. Die stereomikroskopischeUntersuchung aufgehellter Schnitte kann zeitraubende Rekonstruktionen ersetzen. Der Nucleus reticularis lateralis wird in neun regelmäßig angelegte Areale unterteilt: Pars principalis, dorsalis, medialis, lateralis, perivagalis, marginalis disseminata und subtrigeminalis, sowie Promontorium und Repagulum cuneati. Zusammen mit dem Nucleus interfascicularis hypoglossi und dem Kern von Roller tragen die Unterkerne des Nucleus reticularis lateralis zahlreiche gemeinsame Züge, so daß sie als eine morphologisch und wahrscheinlich auch funktionell zusammengehörige Gruppe beschrieben werden können. Die Studie weicht in der Nomenklatur und den Abgrenzungen der einzelnen Areale in zahlreichen Einzelheiten von den Darstellungen anderer Autoren ab. Diese Unterschiede werden diskutiert.
    Notes: Summary The three dimensional shape of the nucleus reticularis lateralis, nucleus interfascicularis hypoglossi, the nucleus of Roller and a hitherto unknown cellular layer (Repagulum cuneati) is described on the basis of 800 μ sections, which are stained with aldehydefuchsin. By this method lipofuscin granules in the nerve cells are stained selectively. The stereomicroscopical investigation of the sections can replace time consuming reconstructions. The nucleus reticularis lateralis is divided into nine constantly occuring areas: pars principalis, dorsalis, medialis, lateralis, perivagalis, marginalis disseminata and subtrigeminalis, promontorium and repagulum cuneati. Together with the nucleus interfascicularis hypoglossi and the nucleus of Roller the subnuclei of the nucleus reticularis lateralis have many features in common indicating them to represent a morphological and presumably a functional unit. The present study is at variance in numerous details with the conceptions of other authors concerning the nomenclature and delineation of the nuclei. These differences are discussed.
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  • 18
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    Cell & tissue research 131 (1972), S. 235-254 
    ISSN: 1432-0878
    Keywords: Cortex cerebri ; Human ; Ammonsformation ; Subiculum ; Pigmentarehitecture ; Lipofuscin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung Mit Hilfe einer neu entwickelten Methode zur Darstellung von Neurolipofuscinen werden die am Aufbau des Subiculum beteiligten Zellschichten elektiv hervorgehoben. Da sich die Methode auf die Darstellung nur einer Cytoplasmakomponente beschränkt, werden Unterschiede zwischen den verschiedenen Nervenzelltypen stärker betont als im Nisslbild. Weiterhin lassen sich bis zu 800 μ dicke Schnitte verwenden die — unter dem Stereomikroskop betrachtet — die Verfolgung und Grenzbestimmung der einzelnen Zellschichten außerordentlich erleichtern. Im Pigmentbild stellt sich das Subiculum nicht als eine Übergangsrinde zwischen dem Cornu ammonis und den entorhinalen Feldern dar, sondern erscheint als eine eigene Region innerhalb der Ammonsformation mit kennzeichnenden Zellschichten, die in den benachbarten Feldern nicht vorkommen. Diese eigenständigen Schichten werden durch unmittelbar aufeinanderfolgende Lagen großer und mittelgroßer Pyramidenzellen gebildet (Lamina pyramidalis externa und interna subiculi). In den großen Subiculumneuronen des äußeren Blattes finden sich neben Pigmentanhäufungen im apikalen Cytoplasma vor allem bemerkenswerte Lipofuscinkonzentrationen in den mittleren Abschnitten der Spitzendendriten, ein Verteilungsmuster, das im Zentralnervensystem des Menschen nur selten angetroffen wird. Die Neurone der Lamina interna speichern demgegenüber das Lipofuscin nur im Zelleib. Die beiden eigenständigen Subiculumschichten werden durch das Eindringen fremder Zellschichten aus der Umgebung ergänzt. Zellwolken aus dem Praesubiculum schieben sich in der Lamina zonalis weit in den Subiculumbereich vor. Außerdem wird ein Randabschnitt des Subiculum von Abkömmlingen der inneren Hauptschicht aus der Regio entorhinalis unterlagert (Pri-β und Pri-γ). Vom Sektor CA I (h1) her schieben sich Ammonspyramiden über die Lamina pyramidalis externa subiculi. Alle Schichten verändern kontinuierlich ihre Breite, so daß die Zusammensetzung der Rinde örtlich großen Veränderungen unterworfen ist und kein Punkt innerhalb dieser Region einem anderen gleicht. Das Subiculum des Menschen besteht aus einem langgestreckten flügelförmigen Areal zwischen Cornu ammonis und Regio entorhinalis, bzw. praesubicularis, und einem schalenförmigen Abschnitt im Gyrus uncinatus. Beide sind an ihrem rostromedialen Pol miteinander verbunden. In diesem Areal grenzen wir mit Hilfe der Pigmentarchitektonik acht Felder gegeneinander ab. Im Bereich des sechsschichtigen s med.oral. werden die beiden eigenständigen Zellblätter von Zellwolken und -platten aus dem Praesubiculum und den Schichten Pri-β und Pri-γ aus der Regio entorhinalis zangenartig eingefaßt. Die Zahl der Schichten wird in s med.caud. durch das Verschwinden der entorhinalen Zellamellen reduziert. Das seitliche Feld ist dadurch gekennzeichnet, daß Neurone des Cornu ammonis in wechselnder Breite die Lamina pyramidalis externa subiculi überdecken. Da die äußere Schicht der großen Subiculumneurone nicht so weit nach lateral reicht wie die innere Schicht, läßt sich ein großflächiges vierschichtiges Zentralfeld (s lat.centr. ) von einem schmalen dreischichtigen Randstreifen (s lat.marg. ) unterscheiden. Im Gyrus uncinatus wiederholen sich die genannten Felder in leicht abgewandelter Form. Dementsprechend findet sich ein mediales Feld (s g.u.med. ) mit Praesubiculuminseln und Pri-γ und ein laterales Areal (s g.u.lat.centr.,marg. ) mit CA-I-Neuronen. In Richtung auf den Limbus Giacomini ist eine weitere weniger hoch differenzierte vierschichtige Randzone entwickelt (s g.u.dors. ), die aus Zellmaterial des Gyrus intralimbicus und des Subiculum besteht.
    Notes: Summary By means of a newly developed method demonstrating neurolipofuscines the cellular layers constituting the subiculum are stained selectively. Since this method is confined to the demonstration of one cytoplasmic component, differences between various types of neurons show up more clearly than in Nisslpreparations. In addition up to 800 μ thick sections can be used which—if viewed at under the stereomicroscope—facilitate the recognition and identification of borders of cortical cellular layers. Pigmentarchitecturally the subiculum does not appear as a transitory zone between ammonsformation and entorhinal areas, but represents an own region within the ammonsformation with characteristic cellular layers which are absent in neighbouring fields. These autochthonous layers are constituted by large and medium sized pyramidal cells (lamina pyramidalis externa and interna subiculi), which lie closely together. In the large subicular neurons of the outer lamina beside pigment accumulations in the apical cytoplasm, above all remarkable concentrations of lipofuscin in the middle parts of the apical dendrites are to be found. This pattern of distribution is only rarely to be encountered in the central nervous system of man. The neurons of the lamina interna on the contrary store their lipofuscin in the perikaryon. The two autochthonous subicular layers are completed by the invasion of foreign cellular layers from the surrounding fields. Cell islands from the praesubiculum penetrate far into the lamina zonalis of the subiculum. Further on a marginal part of the subiculum is underlain by derivatives of the inner principal layer of the regio entorhinalis (Pri-β and Pri-γ). Ammonspyramids from CA I (h1) overlie the lamina pyramidalis externa subiculi. All layers change continuously their width, so that the appearance of the cortex is submitted to extensive local alterations. The subiculum of man consists of an elongated wingshaped area between cornu ammonis and regio entorhinalis, or praesubiculum respectively, and a cupshaped part in the gyrus uncinatus. Both are connected at their rostromedial poles. In this region on the basis of the pigmentarchitecture eight areas are to be separated. In the six-layered s med.oral. the two autochthonous cellular layers are embraced by cell-clouds and -plates from the praesubiculum and the layers Pri-β and Pri-γ from the regio entorhinalis. The number of the layers in the s med.caud. is reduced by the disappearance of the entorhinal laminae. The lateral field is characterised by the fact that the lamina pyramidalis externa subiculi is covered in various extent by neurons of the cornu ammonis. Since the outer layer of the large subiculum neurons does not extend so far laterally as the inner layer, a large four-layered central field (s lat.centr. ) can be distinguished from a narrow three-layered marginal strip (s lat.marg. ). In the gyrus uncinatus the same fields can be found, however in a slightly modified form. Thus a medial field (s g.u.med. ) with presubicular islands and Pri-γ and a lateral area (s g.u.lat.centr.,marg. ) with CA I neurons are to be detected. In the direction of the limbus Giacomini a further less highly differentiated four-layered marginal area (s g.u.dors. ) is developed which consists of material of the gyrus intralimbicus and the subiculum.
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  • 19
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    Cell & tissue research 135 (1972), S. 415-438 
    ISSN: 1432-0878
    Keywords: Brain stem ; Human ; Dorsal glossopharyngeus vagus-complex ; Lipofuscin ; Light microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung Mit Hilfe einer neu entwickelten Methode zur Darstellung von Neurolipofuscinen wird das Pigmentbild des dorsalen Glossopharyngeus- und Vagusgebietes beschrieben. Da außer wenigen elastischen Fasern nur die Lipofuscingranula, welche in Nervenzellen gespeichert sind, angefärbt werden, können Schnitte bis zu einer Dicke von 800 μ verwendet werden. Sie lassen bei stereomikroskopischer Betrachtung die Grenzen von Kerngebieten meist deutlich erkennen, da durch die Übereinanderprojektion zahlreicher Neurone auch geringe Unterschiede in der Dichte der Zellen, sowie in der Art und dem Ausmaß ihrer Pigmentierung hervortreten. Das dorsale Glossopharyngeus- und Vagusgebiet wird in elf Kerne unterteilt. Seinen caudalen Pol bilden der Nucleus terminalis caudalis alae cinereae, der Nucleus commissuralis und parvocellularis. Im mittleren Bereich liegen zahlreiche Kerne von kennzeichnender Gestalt nahe beieinander. Der Nucleus pigmentosus nervi vagi hüllt das Gesamtgebiet ein und füllt die Räume zwischen den Kernen aus. Der Nucleus alaris nervi vagi durchzieht als breite Zellplatte den ganzen mittleren Teil. Seine großen Neurone speichern Lipofuscinkörnchen nicht nur in den Perikaryen sondern auch in Form langer Pigmentspindeln in den Dendriten. Im kleinzelligen Gebiet lateral vom alaren Kern werden ein Nucleus terminalis intermedius alae cinereae, der Kern der Area postrema, ein Nucleus tractus solitarii, ein Nucleus gelatinosus, sowie ein oraler Terminalkern der Ala cinerea voneinander unterschieden. In allen Terminalkernen findet sich reichlich Dendritenpigment neben den kräftig gefärbten Lipofuscinkegeln in den Zelleibern. An der Grenze zum oralen Abschnitt des Gesamtgebietes vermindert sich die Dichte der Pigmentierung. Hier liegt in der Fortsetzung des alaren Vaguskernes der Nucleus salivatorius inferior. Eine so weitgehende Untergliederung des Glossopharyngeus- und Vagusgebietes steht in Widerspruch zu den Darstellungen zahlreicher Autoren. Vor allem im kleinzelligen Bereich, der meist als ein einheitliches Areal aufgefaßt wird, können mit Hilfe der Pigmentarchitektonik die drei Terminalkerne der Ala cinerea eindeutig vom eigentlichen Kern des Solitärbündels und vom Nucleus gelatinosus unterschieden werden. Außerdem stellt sich heraus, daß die Gesamtheit der melaninhaltigen Neurone im Vagusgebiet, zusammen mit einem weiteren großen Zelltyp, der reichlich mit Lipofuscin beladen ist, ein eigenes kompliziert geformtes Kerngebiet bilden, das sicher von den übrigen Kernen, auch von dem gleichfalls großzelligen Nucleus alaris abgetrennt werden kann.
    Notes: Summary By means of a newly developed method for the demonstration of neurolipofuscines the distribution of pigments in the dorsal glossopharyngeus-vagus-complex is described. Since apart from few elastic fibers only the lipofuscin granules which are stored in neurons are stained, sections up to 800 μ can be investigated. Under the stereomicroscope they show in general clearly the borders of nuclei, since because of the superposition of numerous neurons also slight differences in respect of the density of the cells and type and extent of their pigmentation show up clearly. The dorsal glossopharyngeus-vagus-complex of the human is subdivided into eleven nuclei. Its caudal pole is formed by the nucleus terminalis caudalis alae cinereae, the nucleus commissuralis and parvocellularis. In the middle portion several nuclei of characteristic shape lie closely together. The nucleus pigmentosus nervi vagi envelopes the whole complex and fills the spaces between the nuclei. The nucleus alaris nervi vagi forms a broad plate of cells extending over the whole length of the middle portion. Its large neurons do not only store the lipofuscin granules in their perikarya but also in form of elongated pigment spindles in the dendrites. In the parvicellular parts lateral of the nucleus alaris a nucleus terminalis intermedius alae cinereae, a nucleus of the area postrema, a nucleus tractus solitarii, a nucleus gelatinosus, and a nucleus terminalis oralis alae cinereae are to be distinguished. In all terminal nuclei abundant pigment spindles of the dendrites occur beside the heavily stained lipofuscin pyramids in the perikarya. At the border of the oral part of the complex the density of pigmentation decreases. Here, in continuity of the nucleus alaris, the nucleus salivatorius inferior is situated. Such an extensive subdivision of the glossopharyngeus — vagus-complex of the human is in contrast to the description of numerous authors. Especially in the parvicellular part, which generally is described as a uniform area, by means of pigment architecture unequivocally the three terminal nuclei of the ala cinerea can be distinguished from the nucleus tractus solitarii proper and the nucleus gelatinosus. In addition it becomes apparent by the pigment architecture that the whole population of melanin containing neurons in the vagus-complex, together with a further large cell type, which stores vast amounts of lipofuscin granules, forms a distinctive nucleus which can be separated with certainty from the other nuclei, as well as the equally magnocellular nucleus alaris.
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  • 20
    ISSN: 1432-0878
    Keywords: Fourth ventricle ; Human ; Ependyma ; Scanning electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Scanning electron microscopy was used to assess the ultrastructural differences exhibited by the varigated ependymal lining of the near-term human fetal 4th ventricle. The central portion of the fourth ventricular floor, including the median sulcus is punctuated by numerous clumps of cilia. The density of cilia here is not as great as that described for other regions of the human cerebral ventricular system; accordingly, underlying substructure can be noted. There are distinct differences between ependymas that line the floor of the fourth ventricle with those of the adjacent area postrema. The latter region possesses not cilia, but instead exhibits a dense knap of microvilli. The ultra-architecture of the choroid plexus is relatively similar to that of other circumventricular organs with the exception that it possesses small isolated groups of cilia as well as microvilli. These findings are discussed with respect to the dynamics of local CSF movement and flow, ependymoabsorption and ependymosecretion
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  • 21
    ISSN: 1432-0878
    Keywords: Key words Apoptosis ; p53 ; Ischemia ; Enterocytes ; Proliferation ; Differentiation ; ISEL ; Glomeruli ; Mouse (Balb/c) ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Morphological changes associated with apoptosis are closely correlated with the expression of specific proteins. However, the cause-effect relationships between the expression of these proteins and DNA degradation are barely known. For studying expression of apoptosis-related proteins in relation to different degrees of DNA fragmentation, the small intestine with its spatially organized continuum of proliferation, differentiation and death is a very useful preparation. Enterocytes towards the apex of the villi become increasingly susceptible to apoptosis. Here, this ”apoptotic gradient” is used to demonstrate the presence of BAX and BCL-2 proteins in the cytoplasm of cells at the onset of apoptosis. In semithin serial sections of the small intestine, BAX, BCL-2 and DNA fragmentation were demonstrated. BAX and BCL-2 are always colocalized and only in cells with fragmented DNA. The gradient of BAX or BCL-2 staining is similar to the gradient of DNA fragmentation. Immunoreactivity for BCL-2 or BAX is most intense in cells that are prone to become apoptotic next in the course of cellular turnover but not in cells in an advanced apoptotic state, showing strongly condensed chromatin. When using the same technique on semithin sections of kidney biopsies, containing epithelia with low cellular turnover, we found DNA fragmentation mainly in the epithelial cells of the distal tubules. Similar to the situation in the enterocytes, BAX staining was confined to the cytoplasm of epithelial cells with a moderate degree of DNA fragmentation and reduced in epithelial cells with a high degree of DNA fragmentation. In contrast to the situation in the small intestine, very low levels of BCL-2 were found. The results suggest that expression of BCL-2 and BAX is related to cell damage as indicated by DNA fragmentation but not to advanced stages of cellular death, as indicated by chromatin condensation and cellular shrinkage.
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  • 22
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    Cell & tissue research 139 (1973), S. 333-350 
    ISSN: 1432-0878
    Keywords: Placenta ; Human ; Blood vessels ; Endothelium ; Vascular muscle, Myoendothelial junctions ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung Die Gefäße der Nabelschnur, Chorionplatte, Stammzotten und das Mikrozirkulationssystem des Zottenbaums der reifen menschlichen Plazenta wurden elektronenmikroskopisch untersucht. Die Endothel- und Muskelzellen besitzen auch am Ende der Gravidität alle morphologischen Kennzeichen voller Funktionsfähigkeit. Auffällig sind die zahlreichen myo-endothelialen Verzahnungen und die zahlreichen Kontakte der Muskelzellen untereinander. Sphinkterartige Einrichtungen werden nur an den Verzweigungen der Präkapillaren beobachtet. Gefäßnerven fehlen überall. Möglicherweise kann die Durchströmung der fetalen Placentagefäße durch autonome Kontraktion der Gefäßwände reguliert werden. — Alle Gefäße der Placenta sind elasticafrei. In den Nabelarterien sind elastische Systeme schwach ausgebildet. Eine Elastica interna wird nur in der Nabelvene gefunden. — Das Mikrozirkulationssystem in den Rami und Ramuli chorii sowie den Terminalzotten wird von allen bekannten Endstromeinheiten aufgebaut. Die englumigen Kapillarabschnitte befinden sich vor allem in unmittelbarer Nachbarschaft zum organellenreichen Syncytiophoblast, während die weitlumigen Abschnitte, die möglicherweise die venösen Kapillarstrecken sind, den Epithelplatten anliegen.
    Notes: Summary In the human placenta at term the blood vessels of the umbilical cord, chorionic plate and trunks and the microcirculation system of the placental lobes have been investigated. Also at the end of pregnancy the vascular endothelium and the smooth muscle cells exhibit all morphological criteria of full functional activity. The vessel wall is characterized by numerous myoendothelial junctions and many adjacent muscle cells being in close contact with one another. Sphincterlike structures can only be observed at the ramifications of the precapillaries. Vascular nerves are always absent. The blood circulation is perhaps regulated autonomously by means of the contractile vessel wall.—In all placental vessels a special tunica elastica never exists. In the umbilical arteries elastic systems are poorly developed. A genuine internal elastic layer only occurs in the corresponding vein.—In the region of the rami and ramuli chorii as well as in the single villi the microcirculation system consists of all types of terminal vessels. The capillaries with small diameter are preferentially situated close to the organellrich syncytiotrophoblast, whereas the distended segments, which may represent the venous capillaries, are in intimate contact with the epithelial plates.
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  • 23
    ISSN: 1432-0878
    Keywords: Key words Vesicle protein ; Physin ; Secretory carrier-associated membrane protein ; Vesicle-associated membrane protein ; Synaptobrevin ; Expression ; Liver ; Bovine ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The cell-type restricted expression of cytoplasmic microvesicle membrane protein isoforms may be a consequence of the functional adaptation of these vesicles to the execution of specialized processes in cells of different specialization. To characterize the expression of the vesicle protein pantophysin, an isoform of the synaptic vesicle proteins synaptophysin and synaptoporin, we have prepared and characterized antibodies useful for the immunological detection of pantophysin in vitro and in situ. Using these reagents, we show by immunoblot analyses that pantophysin expression is not homogeneous but differs significantly between various bovine tissues. Furthermore, these differences are not exactly paralleled by the expression of other vesicle proteins of the SCAMP (secretory carrier-associated membrane protein) and VAMP (vesicle-associated membrane protein) types that have previously been localized to pantophysin vesicles in cultured cells. By immunofluorescence microscopy, pantophysin expression is seen predominantly in non-neuroendocrine cells with pronounced membrane traffic. Pantophysin staining codistributes with SCAMP and VAMP immunoreactivities in most instances but differs in some. Remarkably, pantophysin staining in liver is restricted to cells surrounding sinusoids and is not detectable in hepatocytes, similar to that of the SCAMP and VAMP isoforms as detected by our reagents in tissue sections.
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  • 24
    ISSN: 1432-0878
    Keywords: Key words Mammary gland ; Differentiation ; Neoplasia ; Homeodomain ; Cancer ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  In the mouse mammary gland, homeobox gene expression patterns suggest roles in development and neoplasia. In the human breast, we now identify a family of Iroquois-class (IRX) homeobox genes. One gene, IRX-2, is expressed in discrete epithelial cell lineages being found in ductal and lobular epithelium, but not in myoepithelium. Expression is absent from associated mesenchymal adipose stroma. During gland development, expression is concentrated in terminal end buds and terminal lobules and is reduced in a subset of epithelial cells during lactation. In contrast to observations for many homeobox genes in the mouse mammary gland in which homeobox gene expression is lost on neoplastic progression, IRX-2 expression is maintained in human mammary neoplasias. Data suggest IRX-2 functions in epithelial cell differentiation and demonstrate regulated expression during ductal and lobular proliferation as well as lactation.
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  • 25
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    Cell & tissue research 163 (1975), S. 133-150 
    ISSN: 1432-0878
    Keywords: Cerebral capillaries ; Human ; Development in vitro
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The use of an organotypic-in the strictly literal meaning of the word, nervous tissue culture device has allowed the identification and ultrastructural study of various types of developing capillaries in human cerebellum and olfactory bulb in vitro. Most capillaries were similar to those already described by other authors or by us, in human or animal embryos and fetuses. Large Type I Capillaries Their luminal diameters were greater than 8 microns. The basement membranes were thin and discontinuous. Numerous interendothelial junctions were either plate-like attachments or contained pentalaminar zones. Type II Capillaries Their lumina were between 2 and 8 microns in diameter. The basement membranes were wider than those of type I capillaries and were sometimes continuous. The interendothelial junctional complexes of type II capillaries included pentalaminar portions. Many simple or complex vascular sprouts (type IV and V capillaries) had small or non-patent lumina. Their basement membranes were absent or very thin and discontinuous. Their interendothelial junctions were similar to those of type I capillaries. Some of the less frequently encountered capillary types seen in developing human nervous tissue were absent in culture. Some pathological features were seen—especially in long-term cultures—in type I and II capillaries containing degenerating blood cells or processes sometimes obviously related to histiocytic cells. They consisted mainly of an accumulation of microfilaments and modifications of the rough endoplasmic reticulum in the endothelial cells. These pathological changes did not modify the main characteristics of the capillaries. The origin of the vascular sprouts, the exact nature of the interendothelial junctions and the significance of the pathological changes are discussed. This model may prove useful for the study of cerebral vasculogenesis, the development of the blood-brain barrier and the physiological or pathological properties of the human brain capillaries in tissue culture.
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  • 26
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    Cell & tissue research 175 (1976), S. 1-15 
    ISSN: 1432-0878
    Keywords: Rete testis ; Human ; Epithelial cell types ; Smooth muscle cells ; Histophysiology ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructure of the normal human rete testis was analyzed. The rete testis cavities are irregularly shaped and contain virtually no spermatozoa. Smooth muscle cells often surround the cavities. In the epithelial lining, two cell types are distinguishable. Flat, dark cells exhibit numerous slender microvilli, and numerous apical and basal microvesicles. Prismatic, lighter cells have more cell organelles, mostly polarized towards a supranuclear position. Both cell types contain variable amounts of glycogen and fat, and an occasional cilium. All cells display intricate lateral cell surfaces that possess different cell-to-cell attachment devices. Intermediate cell types are frequently found. On a morphological basis, the epithelial cells seem to be involved in the release of substances into the lumen and probably also in transport towards the base. Connective tissue elements are found subjacent to the epithelium. Scattered among the fibrocytes are typical smooth muscle cells. Expansions of some smooth muscle cells are connected to the epithelial basement membrane by a network of microfibrillar material. The smooth muscle cells may be involved in changing the shape of the rete testis channels, thus promoting the flux of the rete testis fluid. Different types of nerve fibre bundles are distinguished in the connective tissue of the rete testis which may correspond to autonomic and sensory nerves or sensory receptors.
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  • 27
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    Cell & tissue research 168 (1976), S. 247-259 
    ISSN: 1432-0878
    Keywords: Tendons ; Connective tissue ; Collagen ; Human ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A variety of human tendons have been studied at the electron microscope level. The fibers of these tendons are composed of collagen fibrils that average 1,750 Å and 600 Å in diameter. A third population that measures 100 Å in diameter may represent immature collagen or filaments that are incorporated into tendon elastic fibers. The larger collagen fibrils vary in ratio with respect to one another, and are connected by interfibrillar bridges which in some cases appear to extend through the substance of the fibril. The collagen fibrils of the paratenon are less-well organized than those of the tendon proper and average 600 Å in diameter. Tendons that exhibit the property of lateral stretch (plantaris and palmaris) were compared at the ultrastructural level with tendons that do not have this property. No differences between the two tendon types could be determined in normal or spread preparations, indicating that the differences in physical characteristics are a result of fiber rather than fibril organization.
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  • 28
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    Cell & tissue research 109 (1970), S. 356-379 
    ISSN: 1432-0878
    Keywords: Gingivo-Dental junction ; Human ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Résumé L'étude ultrastructurale de la jonction gingivo-dentaire humaine a permis de montrer que les membranes cytoplasmiques des cellules épithéliales superficielles, revêtues de nombreux hémi-desmosomes, sont en rapport avec les cristaux d'apatite de l'émail par l'intermédiaire d'un espace intercellulaire, de 400 à 1200 Å d'épaisseur, rempli d'un matériel finement granulaire, soit directement soit par interposition d'une cuticule discontinue, d'épaisseur variable, qui est appliquée à la surface de l'émail uniquement sur des dents, ayant effectué leur éruption. Cette cuticule, dont l'origine salivaire peut être exclue, subit une apposition de la surface de l'émail vers la périphérie. L'attache épithéliale n'est pas kératinisée, mais des faisceaux de tonofibrilles sont visibles dans les cellules. Une certaine tendance à la kératinisation a été parfois notée au niveau du sillon gingivo-dentaire. De nombreux polynucléaires éosinophiles et neutrophiles et des lymphocytes migrent à travers la jonction et le sillon gingivo-dentaires cliniquement normaux. Un élargissement des espaces intercellulaires, des ruptures de la membrane basale et des desmosomes, une accumulation parfois importante de glycogène, des noyaux polylobés et des oedèmes intracellulaires y ont également été notés. Enfin une rupture en ≪atimbre poste≫ entre les cellules épithéliales de la jonction, ainsi qu'une rupture entre l'émail et les cellules épithéliales superficielles contribuent à l'approfondissement du sillon gingivo-dentaire.
    Notes: Summary The ultrastructural investigation of the human gingivo-dental junction shows that the plasmalemmata of the superficial epithelial cells, coated by numerous hemi-desmosomes, are related to the enamel apatite crystals through a finely granular intercellular space, 400 to 1200 Å in thickness, either directly or through a discontinuous dense cuticle, of irregular thickness, which is attached to the enamel surface of erupted teeth. This cuticle, of none salivary origin, is deposited from the enamel surface to the outside. The epithelial junction is not keratinized, but numerous tonofibrils bundles occur in the cells. A certain tendency toward keratinization was sometimes observed in the crevicular epithelium. Numerous eosinophilic and neutrophilic polymorphonuclear leukocytes, and lymphocytes migrate through the clinically normal crevicular and junctionnal epithelium. Intercellular space dilatations, basement membrane and desmosomes ruptures, important intracellular accumulations of glycogen, multilobulated nuclei and intracellular edemas are also sometimes observed. A serrated rupture between epithelial cells of the junction as well as a rupture between enamel and the most superficial epithelial cells contributes to the deepening of the gingival sulcus.
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    Cell & tissue research 109 (1970), S. 306-315 
    ISSN: 1432-0878
    Keywords: Eye ; Human ; Iris ; Senile Cataract ; Extracellular components
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung Die extrazellulären Komponenten in der Iris von vier menschlichen Augen mit Altersstar wurden untersucht. Folgende Befunde wurden erhoben: 1. Die hintere Irisfläche ist von einer teilweise mehrschichtigen Basalmembran bedeckt. 2. Im Irisstroma kommen einige Gruppen von ringförmig angeordneten Kollagenfibrillen vor. 3. Die Interzellulärräume im Spincter- und Dilatatormuskel werden von einer granulären Substanz ausgefüllt, die Filamente, Kollagenfibrillen und einige dichte Körperchen enthält. 4. Im Irisstroma liegt fibrilläres Material mit einem symmetrischen Querstreifungsmuster vor, dessen Perioden etwa 650 Å lang sind. 5. Darüberhinaus wurden im Irisstroma Gruppen von quergestreiften Fibrillen mit einem gleichmäßigen Durchmesser von etwa 130 Å beobachtet. 6. Einige Stromazellen sind teilweise von einer feingranulären Substanz umgeben.
    Notes: Summary The extracellular components in iris of four human eyes with senile cataract were studied. The following results were obtained: 1. The posterior iris surface is covered by a basement membrane, which sometimes appears multilayered. 2. Some circular arrangements of collagen fibrils occurs in the iris stoma. 3. The intercellular spaces in the sphincter and the dilatator muscles are filled with a granular substance containing filaments, collagen fibrils, and some dense bodies. 4. A fibrillar material is observed in the iris stroma showing a symmetric cross-band pattern with periods of about 650 Å in length. 5. Groups of cross-banded fibrils with an even thickness of about 130 Å appear in the iris stroma. 6. Stromal cells are sometimes partially surrounded by a finely granular substance.
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    Cell & tissue research 113 (1971), S. 322-343 
    ISSN: 1432-0878
    Keywords: Exocrine pancreas ; Human ; Release mechanism ; Cilia ; Junctional complex
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung 1. An der exokrinen Pankreaszelle des Menschen lassen sich eine Basalzone (ER, Zellkern), eine Golgi-Zone und ein apikaler Zellpol unterscheiden. Die Zymogengranula entstehen durch Membranabknospung aus dem Golgi-Apparat und werden am apikalen Zellpol gespeichert. Die Lumenseite der exokrinen Zelle ist von dichtstehenden Mikrovilli besetzt (200–300 pro Zelloberfläche), die eine netzig-fibrilläre Längsstruktur zeigen. 2. Als morphologisches Substrat für den Vorgang der Extrusion werden wenige elektronendichte Granula am apikalen Pol beschrieben, die durch Aneinanderlagerung und Verschmelzung der Hüllmembranen Doppelformen von Granula oder kurze Granulaketten bilden. Die Extrusion wird abgeschlossen durch das Verschmelzen der Hüllmembran der einzelnen oder komplexen Granula mit der Plasmamembran, durch Ausbildung einer Öffnung in der Plasmamembran und durch das Ausfließen des Granuluminhaltes. Die Zelloberfläche von exokrinen Zellen mit lebhafter Extrusion zeigt nur spärlichen Besatz mit Mikrovilli. 3. Die exokrinen Zellen innerhalb eines Azinus werden an den apikalen und seitlichen Kontaktflächen durch ein differenziertes Schlußleistennetz (Zonulae occludentes, Z. adhaerentes, Desmosomen) verbunden, Tonofibrillen verlaufen bogenförmig von einer Verschlußzone zur nächsten und strahlen radiär ins Zytoplasma der exokrinen Zellen aus. 4. Die Epithelzellen der Schaltstücke sind durch ähnlich gebaute Haftzonen mit den exokrinen Zellen und untereinander verbunden; ihr Zytoplasma enthält einen kleinen Golgi-Apparat und regelmäßig langgestreckte Zilien (9+2-Muster), die aus einer trichterförmigen Vertiefung der Zelloberfläche entspringen und weit in das Schaltstücklumen hineinragen.
    Notes: Summary 1. The cytoplasm of human exocrine pancreatic cells may be divided into 3 regions: basal (ER, nucleus), Golgi zone and apical pole. The zymogen granules are condensed in the Golgi region and are stored at the apical pole. From the surface of a single exocrine cell 200 to 300 microvilli project into the lumen of the acinus, they contain an internal fibrillar structure oriented in the long-axis of each microvillus. 2. Before being released into the lumen of the acinus the content of zymogen granules becomes electron translucent and the limiting membranes of adjacent granules fuse to form duplex or short interconnected series of granules. The release is terminated by coalescence of the limiting membrane of single or complex granules with the plasmalemma, formation of an opening at the cell surface and extrusion of secretory material. Cells with signs of active release have only few microvilli on their luminal surface. 3. In a single acinus the apical and lateral surfaces of exocrine cells are connected by elaborate junctional complexes (zonulae occludentes, z. adhaerentes, desmosomes), in the adjacent cytoplasm curved bundles of tonofilaments run from one junctional complex to the other and extend also over a long distance into the cytoplasm. 4. The epithelial cells of the intercalated ducts are connected with the exocrine cells by the same pattern of junctional complexes and tonofilaments. Their cytoplasm contains a small Golgi complex and always a single cilium, which extends through a funnelshaped indentation of the cell surface over a long distance (6 to 8 μ) into the lumen of the intercalated duct.
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    Cell & tissue research 115 (1971), S. 114-136 
    ISSN: 1432-0878
    Keywords: Germinal centers ; Tonsils ; Human ; Acid phosphatase ; Electronmicroscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung Mit der von Barka und Anderson (1963) angegebenen Methode wurde elektronenhistochemisch die saure Phosphatase (s. P.) in Keimzentren der menschlichen Tonsille untersucht. Nach mannigfaltigen Abwandlungen der Methode wurden diejenigen Versuchsbedingungen ermittelt, die zu optimalen Reaktionen bei noch genügender Strukturerhaltung führen. Zytoplasmakörper mit s. P.-Reaktion (Lysosomen) ließen sich in sämtlichen Zellarten des Keimzentrums nachweisen. Besonders starke Reaktion wiesen die Phagolysosomen der phagozytierenden Retikulumzellen (Makrophagen) auf. Eindeutige Reaktion war bei pH 5,5–6,0 auch im ER der Germinoblasten, basophilen Stammzellen, Plasmazellen, dendritischen (nichtphagozytierenden) Retikulumzellen und phagozytierenden Retikulumzellen zu finden. Bei pH 6,0 fand sich s. P.-Aktivität an der Außenfläche der Zytomembran von dendritischen Retikulumzellen. Diese Enzymreaktion fiel besonders stark in den Membraneinbuchtungen aus. Sie war auch in dem elektronendichten Material nachweisbar, das die Ausläufer der dendritischen Retikulumzellen häufig umgibt. Diese extrazelluläre Enzymaktivität dürfte in Zusammenhang mit der Anlagerung von Antigenen und Antikörpern an die Zelloberfläche der dendritischen Retikulumzellen stehen.
    Notes: Summary The activity of acid phosphatase has been localized with the aid of the electron microscope in germinal centers of human tonsils according to the method of Barka and Anderson (1963). In order to determine the optimal conditions for fixation and incubation with respect to the maintenance of structural integrity and degree of reaction, several modifications of the method were undertaken. Cytoplasmic bodies with reaction products (lysosomes) could be demonstrated within all cell types of germinal centers. The acid phosphatase reaction in the phagolysosomes of phagocytic reticulum cells (macrophages) was extremly strong. At pH 5.5–6.0, definite reactions were also found in the ER of germinoblasts, germinocytes, basophilic stem cells, plasma cells, dendritic (nonphagocytic) reticulum cells, and phagocytic reticulum cells (macrophages). At pH 6.0 acid phosphatase activity was deposited outside dendritic reticulum cells, i.e. on the surface of their cytomembranes. Within the infoldings of their cytomembrane reaction products were also prominent. The enzyme activity could also be shown in the electron dense material which usually surrounds the branches of the dendritic reticulum cells. The activity of acid phosphatase which appears in the extracellular spaces, may have relation to antigens and antibodies which are attached to the cell surface of the dendritic reticulum cells.
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    Cell & tissue research 115 (1971), S. 461-472 
    ISSN: 1432-0878
    Keywords: Seminiferous Epithelium ; Human ; Germinal cells ; Testis ; Male reproduction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Two variants of a method for determining the quantitative structure of the human seminiferous epithelium are described. Both versions (counting cells in whole tubules and in CA's) utilize specific criteria for the identification of cells, tubules and /or CA's. Both methods utilize the Sertoli cell as a constant with the cell types expressed as numbers of cells per Sertoli cell. One hundred biopsies were quantitated by the tubular method and 100 by the CA method. Counts were found to be reproducible and consistent. Comparisons were made between the two methods, different individuals doing the quantitation, quantitations of the right and left testes of the same subject and between biopsies from the same subject taken at different times. No difference was found between the right and left testes or biopsies from the same subject taken at different times. Quantitation of biopsies from one individual before and after testicular X-ray irradiation show how the method can be applied.
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    Cell & tissue research 123 (1972), S. 566-571 
    ISSN: 1432-0878
    Keywords: Amnion ; Human ; Uridine diphosphate glucose ; Dehydrogenase ; Cytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung Die Amnien von reifen menschlichen Plazenten wurden auf das Vorkommen der Uridindiphosphatglucose-Dehydrogenase (UDPG-DH), eines Enzyms der Glucuronsäurebildung, untersucht. Dabei konnte sowohl in den Zellen des Amnionepithels als auch in den Fibroblasten des Amnionmesenchyms eine hohe Aktivität festgestellt werden. Die Bedeutung dieses Befundes im Amnion wird im Zusammenhang mit der Stellung der Glucuronsäure im intermediären Stoffwechsel besprochen.
    Notes: Summary In order to trace uridine diphosphate glucose dehydrogenase, an enzyme for the synthesis of glucuronic acid, the amnions of mature human placentas were investigated. High activity could be found in the amniotic epithelium as well as in the fibroblasts of the amniotic mesenchyma. These findings are discussed in connection with the task of glucuronic acid in general biological pathways.
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    Cell & tissue research 122 (1971), S. 479-487 
    ISSN: 1432-0878
    Keywords: Corpus luteum ; Human ; Vacuoles ; Involution ; Autophagy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A description of the atretic human corpus luteum is given. Membrane-limited bodies of varying dimensions, some containing membrane-whorls, are found in the cytoplasm. These bodies are sometimes limited by a double membrane and reaction product for acid phosphatase is observed both between the double membrane and inside these bodies. The plasma membrane projections penetrating the cell at multiple sites are ATP-ase positive. Our results favour the hypothesis that the plasma membrane is involved in the formation of the limiting membrane around autophagic vacuoles. The process of autophagy is discussed in the light of the cellular energy balance and the possible origin of the hydrolytic enzymes involved in the formation of autophagic vacuoles is given.
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    Cell & tissue research 127 (1972), S. 407-438 
    ISSN: 1432-0878
    Keywords: Cortex cerebri ; Human ; Regio entorhinalis ; Lipofuscin ; Pigmentarchitecture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung Mit Hilfe einer neu entwickelten Methode zur Darstellung der Neurolipofuscine werden die am Aufbau der Regio entorhinalis beteiligten Zellschichten elektiv hervorgehoben. Bei einem solchen Vorgehen werden die Unterschiede zwischen den einzelnen Zellarten stärker betont als im Nisslbild, weil nur eine Cytoplasmakomponente dargestellt wird. Diese Beschränkung erlaubt zugleich die Verwendung sehr dicker Schnitte (bis zu 800 μ), die — aufgehellt — unter dem Stereomikroskop analysiert werden. Auf diese Weise lassen sich Verfugungen aneinandergrenzender Rindenregionen und Kantenbildungen einzelner Rindenschichten sicher erfassen. Die Schichten des Allocortex unterscheiden sich im Pigmentbild deutlich von denen des Isocortex. Sie gehen nicht kontinuierlich ineinander über. Die Rinde der Regio entorhinalis läßt sich in eine Lamina principalis externa (Pre) und eine Lamina principalis interna (Pri) gliedern. Die äußere und innere Hauptschicht sind meist durch einen zellarmen Faserstreifen (Lamina dissecans) voneinander getrennt. Beide Schichten lassen sich weiter unterteilen (Pre-α Pre-β, Pre-γ, Pri-α, Pri-β, Pri-γ). In der Regio entorhinalis des Menschen werden 16 Felder pigmentarchitektonisch voneinander unterschieden. Davon bestehen 11 Felder ausschließlich aus allocorticalen Schichten, während die restlichen Areae, welche den Übergang zum Isocortex bilden, aus einer wechselnden Zahl allo- und isocorticaler Zellschichten zusammengesetzt sind. Im Bereich des Gyrus parahippocampalis lassen sich 7 rein allocorticale Felder voneinander abgrenzen. Die Areae gruppieren sich ringartig mit stufenweise abnehmender Organisationshöhe um ein hoch differenziertes Zentrum, das im oralen und lateralen Bezirk der Regio entorhinalis liegt. Das kennzeichnende Merkmal für die zentralen Felder ist eine Aufspaltung von Pri-α in drei Schichten (Pri-αα, Pri-αβ, Pri-αγ). In dem Feld e centr. lat. sind alle drei Unterschichten der Lamina principalis externa enthalten, während in e centr. med. Pre-γ fehlt. Die angrenzenden Felder mit einheitlichem Pri-α lassen sich wieder in Arae mit Pre-γ (e interpol.lat. , e caud. ) und ein Gebiet ohne Pre-γ (e interpol. med. ) gliedern. In den rostralen und medialen Abschnitten verschmelzen Pri-α und Pri-γ zu einer einheitlichen Zellschicht und bilden damit das Feld e oral. . An der Grenze zum Gyrus ambiens in Nähe des Sulcus rhinencephali inferior findet sich ein schmaler Rindenstreifen, in dem die Schichten der Lamina principalis externa nur mangelhaft ausgebildet sind. Diese limitrophe Zone setzt sich nach caudal in das Grenzfeld zum Praesubiculum (e marg. caud. ) fort. Eine ähnliche areale Gradation wie im Gyrus parahippocampalis findet sich auch unter den vier Feldern des Gyrus ambiens. Das am höchsten organisierte Feld (ga centr. ) liegt im caudalen und medialen Abschnitt und ist durch eine dreischichtige Lamina principalis interna und eine deutliche Lamina cellularis profunda ausgezeichnet. Im angrenzenden Feld ga lat. ist Pri-α stark reduziert. In ga oral. findet sich nur eine einschichtige Lamina principalis interna. Der Grenzstreifen zum Mandelkernkomplex (e marg. oral. ) besteht nur aus Teilen der äußeren Hauptschicht. Der breite Übergangsbereich von den rein allocorticalen Feldern der Regio entorhinalis bis zum Isocortex wird in vier Areae unterteilt, in denen allo- und isocorticale Schichten fugenartig ineinandergreifen. Die Stufungen ergeben sich dadurch, daß die einzelnen Zellamellen unterschiedlich weit vordringen. Eine modifizierte äußere Körnerschicht reicht bis in das Feld e trans. med. ; zugleich wird Pre-α in tiefer gelegene Rindenschichten verlagert. An der Grenze zu e trans. intermed. endet Pre-β. Die Spindelzellschicht beteiligt sich als ein weiteres isocorticales Element am Aufbau des intermediären Übergangsfeldes. Die seitlichen Kanten von Pre-γ und Pri-α bilden die lineare Grenze zum lateralen Übergangsfeld, e trans. lat. , dessen Struktur durch das Hinzutreten einer äußeren und inneren Pyramidenschicht bereits weitgehend dem Isocortex gleicht. Im Feld e trans. caud. findet sich sowohl die Spindelzellschicht als auch Pre-β. Es bildet damit eine Stufung zwischen dem medialen und intermediären Übergangsfeld, die jedoch nur im caudalen Abschnitt der Regio entorhinalis am Übergang zum Praesubiculum vorhanden ist.
    Notes: Summary By means of a newly developed method demonstrating neurolipofuscines the cellular layers constituting the regio entorhinalis are stained selectively. The differences between the individual cell types show up more clearly than in ordinary Nissl-preparations since by the new technique only one cytoplasmic component is stained. This limitation allows at the same time to use rather thick sections (up to 800 μ), which — after clearing — are studied under the stereoscopic microscope. Thus indentations of neighbouring regions of the cortex and the edgelike formations of individual cortical layers can be demonstrated with certainty. The pigmentarchitecture of the allocortical layers differs clearly from that of the isocortex. The layers of the allocortex are not continuous with those of the isocortex. Within the regio entorhinalis the cortex can be divided into a lamina principalis externa (Pre) and a lamina principalis interna (Pri), which are separated by a narrow zone of fibers (lamina dissecans). The two main layers can be further subdivided (Pre-α, Pre-β, Pre-γ, Pri-α, Pri-β, Pri-γ). In the regio entorhinalis of man 16 areas can be distinguished by their pigmentarchitecture. 11 of these areas consist exclusively of allocortical layers, whereas the other areas which form the transitory part to the isocortex consist of various numbers of allo- and isocortical layers. In the region of the gyrus parahippocampalis 7 purely allocortical areas can be separated from each other. These areas are grouped in gradually decreasing levels of organisation round a highly differentiated center, which lies in oral and lateral parts of the regio entorhinalis. The characteristic feature of the central areas is a splitting of Pri-α into three layers (Pri-αα, Pri-αβ. Pri-αγ). The area: e centr. lat. contains all three sublayers of the lamina principalis externa, whereas in e centr. med. Pre-γ is lacking. The neighbouring areas with uniform (not subdivided) Pri-α can again be separated in areas with Pre-γ (e interpol. lat. , e caud. ) and a field without Pre-γ (e interpol. med. ). In the rostral and medial parts Pri-α and Pri-γ fuse forming an uniform cellular layer constituting the area: e oral. . At the border of the gyrus ambiens near the sulcus rhinencephali inferior a narrow strip of cortex is to be found, in which the layers of the lamina principalis externa are only poorly developed. This limitrophic zone continues caudally into the border area to the praesubiculum (e marg.caud. ). A similar areal gradation as in the gyrus parahippocampalis can be found in the four fields of the gyrus ambiens. The area with the highest organisation (ga centr. ) is situated in the caudal and medial part of the gyrus ambiens and is characterised by a three layered lamina principalis interna and a clearly recognisable lamina cellularis profunda. In the neighbouring field ga lat . Pri-α is considerably reduced. In ga oral. only an one layered lamina principalis interna is to be found. The border field to the amygdala (e marg.oral. ) consists only of parts of the lamina principalis externa. The broad transitory region from the exclusively allocortical fields of the regio entorhinalis to the isocortex can be subdivided into four areas, in which allo- and isocortical layers meet in a zone of mutual indentations. The subdivision of the area is based on the different distances of penetration of the individual cellular layers. A modified lamina granularis externa extends into the field e trans. med. ; at the same time Pre-α is translocated into deeper cortical regions. At the border to e trans. intermed. Pre-β terminates. The lamina multiformis (VI) takes part as a further isocortical element in the construction of the area: e trans. intermed. . The lateral edges of Pre-γ and Pri-α form the linear border to the lateral transitory area (e trans. lat. ), the structure of which resembles considerably that of the isocortex by additional appearance of a lamina pyramidalis externa and interna. In the area e trans. caud. a lamina multiformis as well as the cellular layer Pre-β is to be found, thus constituting a gradation between e trans. med. and e trans. intermed. , which, however, is present only in the caudal portions of the regio entorhinalis at the border to the praesubiculum.
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    Cell & tissue research 130 (1972), S. 318-337 
    ISSN: 1432-0878
    Keywords: Tooth ; Human ; Cementum ; Fibres ; Scanning electron microscope
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The morphology of the cementum surface of human teeth was studied directly by scanning electron microscopy of 50 freeze-dried and 170 anorganic specimens. Extrinsic (Sharpey) fibres were found to occupy almost 100% of the surface in acellular cement, about 40% of the surface in cement with intrinsic fibres but no cells, and 15–40% in cellular cement, and averaged 6 μm diameter. Some areas with no extrinsic fibres resembled adult lamellar bone. The mineral front of the intrinsic fibres generally was similar to forming and resting lamellar bone; that of the extrinsic fibres varied according to the activity of the surface, but was normal to the axis of the fibre, even where these entered the surface at a sharp angle. Resorption bays were either small and isolated, often seen in newly erupted teeth; or, more rarely, large and quite deep where excessive force must have occurred.
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    Cell & tissue research 134 (1972), S. 555-563 
    ISSN: 1432-0878
    Keywords: Endocervical epithelium ; Human ; Granulo-filamentary bodies ; Light- and electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung Elektronenmikroskopische Untersuchungen am schleimbildenden Epithel der Endocervix zeigen mit regelmäßiger Häufigkeit einen Umbau reifer Mucingranula zu granulo-filamentär organisierten Körpern. Diese finden sich im Zervixepithel von Feten ab 35 cm Länge sowie bei Frauen in der Geschlechtsreife und im Senium. Eine funktionelle Deutung dieser Umbauvorgänge ist bisher nicht möglich. Ein lichtmikroskopisches Korrelat konnte nicht mit Sicherheit nachgewiesen werden. Der granulo-filamentäre Umbau der Sekretgranula ist offenbar ein irreversibler Vorgang. Die dabei entstehenden Körper bleiben bei der Sekretabgabe im Zytoplasma der Zelle zurück.
    Notes: Summary By means of electron microscopical investigation of the endocervical epithelium a transformation of ripe mucous granules into granulo-filamentary bodies could be frequently shown. These bodies are found in the cervical epithelium of fetus of at least 35 cm length as well as in the cervical epithelium of younger and old women. We have not yet been able to give a functional explanation of this transformation process. It has not been possible to correlate these electron microscopical findings with light microscopical ones. The granulofilamentary transformation of the mucous granules is obviously an irreversible process. Thus produced bodies remain in the cytoplasm of the cell when mucous is extruded.
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    ISSN: 1432-0878
    Keywords: Mammary gland ; Human ; Milk ; Cell culture
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    Notes: Summary This study traced the origin of cells observed in human breast secretion samples obtained during lactation and describes the appearance of these cells following prolonged maintenance in vitro. Human milk contains a large number of single vacuolated foam cells and a small proportion of non-vacuolated epithelial cells in clusters. Foam cells are identified by their large size, the polarity of their cytoplasmic organelles, the variation in number and size of lipid vacuoles and the condensed chromatin of their eccentrically located nucleus. Both cell types originate by exfoliation from the mammary gland. This was established by comparing the structural characteristics of cells isolated from milk with those of the cuboidal cell linings of ducts and alveoli in lactating mammary tissue. Relatively pure populations of foam cells could be established from early lactation samples (3–7 days post/partum) while non-vacuolated epithelial cell clusters were more frequently cultured from late lactation specimens (1–10 days postweaning). Foam cells did not divide and lost cytoplasmic organization during prolonged culture. In contrast, non-vacuolated epithelium in clusters proliferated to form colonies of polygonal cells. These results, which imply that foam cells are an active form of the non-vacuolated mammary cells in clusters, call attention to one system for the study of the complex hormonal interactions necessary to induce and maintain lactation.
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    Cell & tissue research 174 (1976), S. 499-518 
    ISSN: 1432-0878
    Keywords: Fibroblast ; Human ; Transmission electron microscopy ; Scanning electron microscopy ; Aggregation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The different stages during aggregation of diploid human skin fibroblasts have been examined by transmission and scanning electron microscopy. As a result of aggregation, fibroblasts form a complex tissue configuration. Numerous intercellular junctions can be observed, while the cells remain polygonal and do not develop an organised intracellular cytoskeleton. Cell division occurs only rarely. After aggregation, signs of progressive auto-digestion develop. Adhesion to a substrate results in outgrowth of the cells and monolayer formation, even when extensive cell damage had occurred. The morphology of fibroblasts in aggregates and in the monolayers, from which they were derived, is compared and the contribution of the aggregate system to the study of fibroblast behavior is discussed.
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    Cell & tissue research 119 (1971), S. 434-450 
    ISSN: 1432-0878
    Keywords: Epidermis ; Human ; Nuclei ; Age dependent changes ; Cytophotometry ; Karyometry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung Die Bauchhaut von 23 männlichen Versuchspersonen verschiedener Altersgruppen wurde histologisch, histochemisch und karyometrisch untersucht. Mit drei voneinander unabhängigen Methoden (Gallocyaninchromalaun-, Feulgen- und UV-Methode) wurden die mittlere Nukleinsäuremenge und das Volumen der epidermalen Zellkerne bestimmt. Auf diese Weise wurden etwa 6000 Zellkerne photometrisch und karyometrisch gemessen und die Ergebnisse statistisch ausgewertet. Die ermittelten Werte stellen einen Beitrag zur biorheutischen Metamorphose der menschlichen Epidermis dar. Folgende Befunde wurden erhoben. 1. Bei männlichen Individuen über 64 Jahren kommt es zu einer Abnahme der Zellkernvolumina und des mittleren Nukleinsäuregehaltes um ca. 35%. Gleiche Verhältnisse ergeben sich für die Zellkerne von der letzten Reihe des Stratum spinosum im Vergleich zu Zellkernen des Stratum basale in jeder Altersgruppe. Auch hier nehmen die Nukleinsäuremenge und die Kernvolumina um 35% ab. Die Konzentration der Nukleinsäure bleibt im wesentlichen konstant. 2. Zwischen Nukleinsäuremenge und den Kernvolumina der Epidermiszellen besteht eine lineare Korrelation. Diese läßt sich in Form stochastischer Punktionen unter Angaben der Korrelationskoeffizienten ausdrücken. 3. Durch die Bestimmung von Normalwerten und deren biologischer wie auch altersabhängiger Variationen sind quantitative Abgrenzungen gegen pathologische Befunde möglich.
    Notes: Summary The abdominal epidermis of 23 male humans in different age groups was histologically, histochemically and karyometrically examined. This was done using Gallocyaninchromalum, Feulgen and UV methods in order to determine the average nucleic acid content and the volume of the epidermal cell nuclei. 6000 cell nuclei were examined photometrically and karyometrically and the results were statistically evaluated. The data obtained could contribute to the knowledge of age dependent changes. The results are as follows: 1. In male individuals over 65 years of age, there is a reduction of nuclear volume and of the average nucleic acid content of approximately 35%. There also exists a reduction in the nuclear volume and the nucleic acid content from the cells of the stratum basale to the cells of the stratum spinosum of approximately 35% in each age group. The concentration of nucleic acid is for all practical purposes constant. 2. There is a linear correlation between the amount of nucleic acid and the nuclear volume of epidermal cells. This correlation can be calculated in the form of a stochastic function. The latter is defined by calculating its coefficients. 3. Knowing the normal values and their biological and age-dependent variations it is possible to differentiate between normal and pathological findings.
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  • 41
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    Cell & tissue research 133 (1972), S. 47-57 
    ISSN: 1432-0878
    Keywords: Endometrium ; Human ; Granular stroma cells ; Relaxin ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Résumé L'ultrastructure générale des cellules K endométriales est semblable chez l'Homme, le Rat et le Singe. Mais les granulations des cellules K humaines sont plus variées que dans ces deux dernières espèces et ont certains aspects suggérant une fonction catabolique. Etant donné cet aspect catabolique et l'existence connue d'une activité phosphatasique acide de ces cellules contenant de la relaxine, les auteurs suggèrent que les cellules K pourraient être des cellules sécrétantes en involution et ques les cellules sécrétant activement la relaxine pourraient avoir un aspect morphologique quelque peu différent de celui des cellules K.
    Notes: Summary The general ultrastructure of endometrial granular stroma cells is similar in man, rat, and monkey. But the granulations of human granular stroma cells are more various than in these two last species and have some aspects suggesting a catabolic function. With respect to this catabolic aspect and the previously shown acid-phosphatase activity of these relaxin-containing cells the authors suggest that the granular stroma cells could be involutive secretory cells and that the active relaxin-secretory cells may have some different morphological aspect.
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  • 42
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    Cell & tissue research 132 (1972), S. 203-215 
    ISSN: 1432-0878
    Keywords: Third ventricle ; Human ; Ependymal surfaces ; Scanning microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A scanning electron microscopic analysis of the adult human third ventricular wall revealed ultra-architectural differences between dorsal and ventral portions. In the brains of thirteen and sixteen week old human fetuses regional differences in the surface organization of lining ependyma were more sharply defined than those of the adult. Alterations in the luminal surfaces of ependyma may reflect differences in the functional capacity of various ventricular areas. The potential role of certain ependyma (tanycytes) and their putative participation in neuroendocrine events is discussed.
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  • 43
    ISSN: 1432-0878
    Keywords: Key words Cytokine ; ELISA ; FACScan ; Lipids ; Macrophage ; Monocyte ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Diabetes (type I and type II) affects approximately 13 million people in the United States. Delayed and incomplete healing of wounds can be a major problem for diabetic patients. Macrophages are an important cell in the complex process of wound repair representing the major source of cytokines throughout the wound-healing process. Cytokines mediate many of the cellular responses critical to timely wound repair. It has been suggested that diabetes impairs wound healing through disruption of local cytokine production. Our previous in vivo studies in rats demonstrated that diabetes-induced and diet-induced hyperlipidemia cause changes in macrophage phenotype and function (Iacopino 1995; Doxey et al. 1998), suggesting that alterations in macrophage cytokine profiles represent the cellular/molecular mechanism responsible for delayed wound healing. The purpose of this study was to investigate how monocyte maturation/differentiation and cytokine production were altered by serum lipids in an in vitro system using human cells. Commercially prepared purified human monocytes were cultured and exposed to serum lipids. Phenotypic analysis of differentiated macrophages was then performed by flow cytometry and fluorescent microscopy using surface antigens specific for various macrophage subsets. Selected cytokines in conditioned medium were assayed using commercial human enzyme-linked immunosorbent assay (ELISA) kits. We demonstrate that serum lipids cause an increase in monocytic differentiation leading to an inflammatory macrophage phenotype rather than a reparative/proliferative phenotype. We also show that serum lipids cause a generalized decrease in macrophage cytokine production using interleukin-1 beta (IL-1β), tumor necrosis factor alpha (TNF-α), platelet-derived growth factor (PDGF), and transforming growth factor beta 1 (TGF-β1) as marker cytokines. Our present in vitro results using human cells confirm our previous in vivo studies in the rat and support the hypothesis that diabetes-induced hyperlipidemia alters the monocyte differentiation process resulting in changes of macrophage subsets and cytokine release at the wound site, ultimately impairing the wound-healing process.
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  • 44
    ISSN: 1432-0878
    Keywords: Key words Cultured epidermis ; Keratinocyte ; Epithelialization ; Wound healing ; Human ; Mouse (NMRI)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Sheets of cultured allogeneic human keratinocytes have been used for the treatment of burns and chronic leg ulcers but there has been no animal assay for the therapeutic action of these cultures. In order to analyze the effects of frozen cultures of human keratinocytes on wound healing, we have developed such an assay based on the rate of repair of full-thickness skin wounds in immunocompetent NMR1 mice. Reepithelialization of the control wounds, originating from the murine epithelium at the edge of the wound, occurred at a constant rate of advance of 150 µm/day. When frozen cultured human epidermal sheets were thawed at room temperature for 5–10 min and applied to the surface of the wound, the murine epithelium advanced at 267 µm/day. Most wounds treated with frozen cultures completely healed after 10 days, whereas most control wounds required 16 days. The accelerated reepithelialization did not depend on the presence of proliferative human keratinocytes in the frozen cultures. The cultures also promoted early formation of granulation tissue and laminin deposition over the surface of the wound bed. This simple assay should permit quantitative analysis of the effects on healing exerted not only by cultured cells, but also by proteins and small molecules.
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  • 45
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    Cell & tissue research 176 (1977), S. 235-242 
    ISSN: 1432-0878
    Keywords: Annulate lamellae ; Spermatogenesis ; Ultrastructure ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Electron microscopic examination of normal human testicular tissue revealed annulate lamellae (AL) in the cytoplasm of primary spermatocytes and spermatids. AL of primary spermatocytes are encountered in the perinuclear region, parallel to the nuclear envelope and form single or multiple membranous profiles containing numerous annuli (500–600 Å in diameter) frequently associated with a fibrillar electron dense material. Spermatids contain numerous layers of AL either continuous with the nuclear envelope and caudal to the acrosome or peripherally positioned in the cytoplasm. Individual lamellae possess terminal dilations and display continuities with the endoplasmic reticulum. The interlamellar space in spermatid AL is entirely filled with a fine granular electron dense material. Additionally, the break-down of AL in spermatozoan residual bodies is indicated by a dilation of AL cisternae to form vacuoles following the dissolution of pore complexes.
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  • 46
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    Cell & tissue research 189 (1978), S. 409-433 
    ISSN: 1432-0878
    Keywords: Rete testis ; Human ; Histophysiology ; Chordae retis ; Scanning electron microscopy ; Transmission electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The human rete testis was examined with regard to 1) the number and distribution of entrances of seminiferous tubules, 2) the light microscopic topography and 3) details of the passages as revealed by scanning and transmission electron microscopy. In a newborn 1474 entrances were counted, approximately 50 % entering from the right and 50 % from the left of the central long axis. Three major subdivisions of the rete were distinguished and described: a septal (or interlobular) part represented by tubuli recti, a tunical (or mediastinal) part which is a true network of channels, and an extratesticular part characterized by dilatations (up to 3 mm wide) which we have called bullae retis. In SEM, cylindrical strands running from wall to wall in the tunical and extratesticular rete spaces are a prominent feature. We have called these chordae retis. They are covered by epithelium and are 5–40 μm wide and 15 to more than 100 μm long. They contain a peculiar tissue consisting of central myoid cells in a fibroelastic matrix. The smaller chordae are avascular. In the light of these findings the rete is interpreted as a highly complex myoelastic sponge. Its function is discussed.
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  • 47
    ISSN: 1432-0878
    Keywords: Microvascular endothelial cells ; Cell growth ; Extracellular matrix ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We introduce two methods, both of which are based on cellular-extracellular matrix interaction, which will facilitate the study of human microvascular endothelial cells. One method describes the means to obtain a G1 population baseline in human microvasclular endothelial cells. Because of the contribution of the extracellular matrix in endothelial cell growth, synchronization in G1 was possible only after the incorporation of angiostatic levels of heparin and hydrocortisone into the extracellular matrix. In the second method, we demonstrate that selective perturbation of human microvascular endothelial cell-extracellular matrix interactions results in the induction of a transitional growth state, between proliferative and differentiated growth states, in human microvascular endothelial cells. In the functional, microtubule formation assays, transitional growth state endothelial cells display rates that are indermediate between those obtained from differentiated and proliferative endothelial cells. Our results demonstrate the importance of the human microvascular endothelial cell-extracellular matrix interaction in the determination of cellular growth state. Our findings also imply that responsiveness of microvascular endothelial cells to their cellular-extracellular matrix environs is highest during the differentiated growth state.
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  • 48
    ISSN: 1432-0878
    Keywords: Spleen ; Fetus ; Development ; Extracellular matrix ; Immuno-electron microscopy ; Transmission electron microscopy ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution of the extracellular matrix proteins types III pN-collagen and IV collagen, laminin and tenascin was investigated in fetal, infant, and adult human spleens by using immuno-electron microscopy. The presence of type III pN-collagen was assessed by using an antibody against the aminoterminal propeptide of type III procollagen. All the proteins other than type III pN-collagen were found in reticular fibers throughout development. In the white pulp of the fetus aged 16 gestational weeks, only an occasional type III pN-collagen-containing fibril was present, although type III pN-collagen was abundant in the reticular fibers of the red pulp. Conversely, in adults, most of the reticular fibers of the white pulp, but not of the red pulp, were immunoreactive for type III pN-collagen. Ring fibers, the basement membranes of venous sinuses, were well developed in both infant and adult spleens. The first signs of their formation could be seen as a discontinuous basement membrane, which was immunoreactive for type IV collagen, laminin, and tenascin in the fetus aged 20 gestational weeks. Intracytoplasmic immunoreactivity for all the proteins studied was visible in the mesenchymal cells of the fetus aged 16 gestational weeks and in the reticular cells of the older fetuses, which also showed labeling for type IV collagen and laminin in the endothelial cells. The results suggest that proteins of the extracellular matrix are produced by these stationary cells.
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  • 49
    ISSN: 1432-0878
    Keywords: Key words: Placenta ; Stem villi ; Actin isoforms ; Myofibroblasts ; Smooth muscle cells ; Immunohistochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The expression patterns of a variety of cytoskeletal antigens were studied in normal human tissues (placenta, umbilical cord, myometrium, colon, mammary gland, testis, skeletal muscle, myocardium) as well as in abnormal human tissues (palmar fibromatosis, fibrocystic disease of the mammary gland, mammary carcinoma). The immunohistochemical binding patterns of the monoclonal antibody GB 42 were compared to those of commercial antibodies directed against vimentin, desmin, smooth muscle myosin, pan actin, α-smooth muscle actin and γ-smooth muscle actin. Methods applied comprised immunohistochemistry on cryostat sections and paraffin sections. Immunogold immunocytochemistry was performed on Lowicryl sections. The patterns of GB 42-binding were confirmed biochemically by SDS-PAGE and Western-blotting, and quantitative amino acid analysis. Our data suggest that the monoclonal antibody GB 42 recognizes an actin isoform which is identical to, or closely related to, γ-smooth muscle actin. Unlike the commercially available antibody against γ-smooth muscle actin, GB 42 does not cross-react with α-skeletal or α-cardiac actins. The GB 42-antigen is expressed in smooth muscle cells, myoepithelial cells and in later stages of differentiation of myofibroblasts, in all the tissues investigated. Throughout the development of smooth muscle cells and myofibroblasts, the appearance of the GB 42-antigen occurs after the expression of vimentin, desmin and α-smooth muscle actin, but prior to the expression of smooth muscle myosin. GB 42 is a reliable marker for higher stages of differentiation of smooth muscle cells and myofibroblasts.
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  • 50
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    Cell & tissue research 286 (1996), S. 145-153 
    ISSN: 1432-0878
    Keywords: Key words: Cardiac fibroblasts ; Extracellular matrix proteins ; Angiotensin II ; Angiotensin receptor ; Ageing ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Fibrosis makes an important contribution to the pathophysiological events leading to the development of heart failure in ischemic and hypertensive heart disease. Since cardiac fibroblasts are mainly responsible for the synthesis and deposition of the extracellular matrix, we have established a method for isolating and cultivating human cardiac fibroblasts from explanted human hearts. The cell yield was 2.14±0.25×106 cells in five independent isolations and the cell purity was 95–97%, contaminating cells being vascular smooth muscle cells and pericytes. Cultured cells were studied with respect to growth properties, morphology and deposition of components of the extracellular matrix. Isolated cells displayed a differentiated phenotype, including the second passage in culture; they synthesised collagen I, III, IV, fibronectin, vitronectin, tenascin and chondroitin sulphate and expressed an atypical angiotensin receptor. This atypical angiotensin receptor internalised angiotensins II and III but not angiotensin IV in a time-dependent manner. Stimulation of the cells with angiotensins II and III but not with angiotensin IV resulted in a dose-dependent stimulation of DNA synthesis. Co-incubation with the subtype-specific receptor antagonists Losartan and PD 123317 did not prevent the stimulation of DNA synthesis. The further characterisation of this receptor should provide insights into the pathobiochemical events leading to heart failure in hypertension and ischemic heart disease.
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  • 51
    ISSN: 1432-0878
    Keywords: Placenta ; Stem villi ; Actin isoforms ; Myofibroblasts ; Smooth muscle cells ; Immunohisto-chemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The expression patterns of a variety of cytoskeletal antigens were studied in normal human tissues (placenta, umbilical cord, myometrium, colon, mammary gland, testis, skeletal muscle, myocardium) as well as in abnormal human tissues (palmar fibromatosis, fibrocystic disease of the mammary gland, mammary carcinoma). The immunohistochemical binding patterns of the monoclonal antibody GB 42 were compared to those of commerical antibodies directed against vimentin, desmin, smooth muscle myosin, pan actin, α-smooth muscle actin and γ-smooth muscle actin. Methods applied comprised immunohistochemistry on cryostat sections and paraffin sections. Immunogold immunocytochemistry was performed on Lowicryl sections. The patterns of GB 42-binding were confirmed biochemically by SDS-PAGE and Western-blotting, and quantitative amino acid analysis. Our data suggest that the monoclonal antibody GB 42 recognizes an actin isoform which is identical to, or closely related to, γ-smooth muscle actin. Unlike the commercially available antibody against γ-smooth muscle actin, GB 42 does not cross-react with α-skeletal or α-cardiac actins. The GB 42-antigen is expressed in smooth muscle cells, myoepithelial cells and in later stages of differentiation of myofibroblasts, in all the tissues investigated. Throughout the development of smooth muscle cells and myofibroblasts, the appearance of the GB 42-antigen occurs after the expression of vimentin, desmin and α-smooth muscle actin, but prior to the expression of smooth muscle myosin. GB 42 is a reliable marker for higher stages of differentiation of smooth muscle cells and myofibroblasts.
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  • 52
    ISSN: 1432-0878
    Keywords: Key words: Astrocytes ; GDNF ; Human cell cultures ; mRNA ; Neurotrophic factors ; Protein kinase C ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The expression and modulation of mRNA for glial-cell-line-derived neurotrophic factor (GDNF) in human glial cells was investigated. Astrocyte cell cultures were isolated from human fetal brains, characterized by immunocytochemistry and maintained in vitro in conditions of high purity; sister cultures were exposed to protein kinase C (PKC) inhibitors for 20 min. Total RNA was extracted from the cell pellets, reverse-transcribed into cDNA and amplified by the polymerase chain reaction (PCR) with primers specific for GDNF. A reverse-transcription/PCR procedure was also performed on mRNA extracted from human fibroblast and lymphocyte cell lines. Human astrocytes grown in the absence of neurons expressed detectable amounts of mRNA for GDNF but no amplification products were observed in fibroblasts and lymphocytes, thus confirming that GDNF production was cell-type specific. After exposure to PKC inhibitors, a dramatic down-regulation of GDNF mRNA was observed in astrocyte cell cultures. Thus, human astrocytes are constitutively capable of producing GDNF, such trophic activity is restricted to neural cells, and PKC plays key roles in signal pathways that regulate the gene activation and production of GDNF.
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  • 53
    ISSN: 1432-0878
    Keywords: Key words: Ageing ; Skin ; Proteinase ; Wound ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Despite the association of increasing age with chronic wound-healing disorders and an impaired rate of healing of acute cutaneous wounds, the role of matrix metalloproteinases (MMPs) is unknown. To determine the spatial and temporal patterns and activities of MMP-1, -2, -3 and -9, 132 healthy humans aged between 19 and 96 years underwent 4-mm punch biopsies followed by wound excision between day 1 and day 180 post-wounding. Wounds showed an age-related increase in MMP-2 and MMP-9 immunostaining from day 3; this was associated with degradation of gelatin as shown by zymograms and with increased proteinase activity as shown by azocoll assays. Distinct spatial localisations for each MMP were observed: MMP-2 was found in epidermal structures; MMP-9 was observed in inflammatory cells up to day 21; MMP-1 was localised to keratinocytes at the wound margin. Normal old skin showed pro-MMP-2 bands on zymography and increased MMP-2 immunostaining. These results indicate that: (1) intrinsic ageing is associated with the up-regulation of MMPs previously associated with chronic wound healing; (2) wound-tissue proteinases are essentially active up to day 21 postwounding; and (3) intrinsic ageing may predispose to tissue breakdown disorders because of MMP-2 up-regulation in normal skin.
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  • 54
    ISSN: 1432-0878
    Keywords: Key words: Fibroblast-specific antibody ; Fibroblast separation and elimination ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The unwelcome presence of fibroblasts in many cell cultures prevents the long term cultivation of various cell types and work with pure populations. Recently, we described a novel fibroblast-specific monoclonal antibody (MAb AS02) that recognises a membrane-bound antigen. We have now developed a method using the fibroblast-specific MAb AS02 immobilised on goat-anti-mouse-magnetic beads to separate contaminating fibroblasts. An endothelial cell line experimentally contaminated with 5%–50% fibroblasts was successfully purified. Additionally, an endothelial cell line with an initial fibroblast contamination of 1.5% was prepared. A proportion of each preparation was cultured with no separation step being performed, whereas the remainder was cultured after purification with MAb AS02 to exclude the presence of a minor number of fibroblasts (〈0.1%). The proportion of fibroblasts increased up to 38% in the fifth passage of culture without elimination of the low initial fibroblast contamination, whereas in the fraction with the separation step, no fibroblasts were detectable by flow cytometry, even after the fifth passage. We also used the antibody to detect the presence of naturally contaminating fibroblasts in thyrocyte cultures. After cultivation of thyrocyte cultures over five passages, the number of fibroblasts increased dramatically up to 50%–80% of the whole population. Subsequently, we successfully applied the method for complete elimination of naturally contaminating fibroblasts from freshly isolated thyrocyte cultures from enzymatically digested thyroid glands. Thus, MAb AS02 is a fibroblast-specific marker that is a useful tool for the detection and elimination of contaminating fibroblasts. The specificity of MAb AS02 permits the universal application of this antibody for human cell cultures of interest.
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  • 55
    ISSN: 1432-0878
    Keywords: Key words: Placenta ; Trophoblast ; Monoclonal antibodies ; HLA class I ; HLA ; G ; HLA ; C ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Expression of HLA class I molecules in trophoblast cells from various locations in normal human first trimester and term placenta was investigated by immunohistochemistry with a panel of monoclonal antibodies against the heavy chains or complete HLA class I molecules complexed with β2-microglobulin. These reagents were also employed to distinguish between the products of different HLA class I loci. In addition to previously characterized reagents, a novel monoclonal antibody against HLA-A molecules (TÜ155) was used. Various choriocarcinoma and transfected cell lines served as controls for the specificities of the monoclonal antibodies. Cells in close contact with maternal cells, such as invading trophoblast cells and cells of the basal plate, expressed β2-m micro globulin in association with HLA-G and HLA-C heavy chains. These class I heavy chains may also have been present as isolated molecules, although not in each of the cells. In contrast, cells of the chorion laeve exclusively expressed HLA-G, and not HLA-A, -B, or -C antigens. Our data support the often discussed immune protective function and the regulatory function of the HLA-G molecule, during invasion. In addition, by using monoclonal antibodies HCA2 (anti-HLA-A and -G), HC10 (anti-HLA-B and -C), TÜ149 (anti-HLA-B, -C, and some -A alleles), SFR8-B6 (anti-HLA-Bw6 and some -C), LA45 (some HLA-A and -B), TÜ48 (anti-HLA-Bw4 and some -A), and TÜ155 (anti-HLA-A), we show the presence of HLA-C molecules in all extravillous trophoblast cells of the cell columns and in the basal plate; the trophoblast cells of the chorion laeve lack this antigen. The function of this molecule is not clear, although a protective function against natural killer cell activity in the endometrium is postulated.
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  • 56
    ISSN: 1432-0878
    Keywords: Key words: Respiratory tract ; Airways ; Serous cells ; Mucous cells ; Mucin ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. We used nine lectins and three antibodies directed against ABO blood-group antigens in morphological and Western-blot experiments to investigate the source of secretory products of human large airways. In tissue sections, the lectins from Griffonia simplicifolia (type I B4), Dolichos biflorus, and Helix pomatia, and the antibodies to the A, B, and/or H-antigen bound to mucous gland cells and to goblet cells; the binding of these substances was dependent on secretor status and ABO blood group. The lectins from Arachis hypogaea, Lens tetragonolobus, Ulex europaeus (type I), Triticum vulgaris, and Sambucus nigra bound to these cell types, regardless of ABO blood group. Serous cells of the tracheal and bronchial glands were stained by the lectins from Canavalia ensiformis, T. vulgaris, Lens tetragonolobus, S. nigra, and U. europaeus (type I). On Western blots of bronchial proteins, the mucins in the high molecular weight region exhibited the same lectin and antibody binding as the mucous gland cells and the goblet cells in the histochemical preparations. The low molecular weight bands were characterized by similar lectin- and antibody-binding properties as the serous gland cells. Thus, mature mucins in the large airways are produced only in the mucous cells of the glands and in the goblet cells, whereas fully glycosylated low molecular weight glycoproteins originate only from the serous cells of the glands.
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  • 57
    ISSN: 1432-0878
    Keywords: Key words: Autoantibodies ; Human anti-Jo-1 antibodies ; Myositis ; Confocal microscopy ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The present study was designed to determine the subcellular localization of histidyl-tRNA synthetase (Jo-1) in human laryngeal epithelial carcinoma cell line (HEp-2 cells). Indirect immunofluorescence using commercial HEp-2 cells with human serum and human-affinity-purified anti-Jo-1 antibodies was performed using confocal microscopy. Anti-histidyl-tRNA-synthetase-positive sera showed distinct nuclear and cytoplasmic granular staining in HEp-2 cells. Affinity purified anti-Jo-1 produced an identical pattern to the whole serum, whereas the serum fraction that did not bind to the affinity column was negative by immunofluorescence on HEp-2 cells. Two commercial human anti-Jo-1-positive control sera and seven anti-Jo-1-positive sera from patients with myositis reproduced the nuclear and cytoplasmic granular pattern. We conclude that Jo-1 is present in cytoplasm and in intact nuclei from HEp-2 cells. The presence of tRNA synthetases in intact nuclei suggests that they have an unsuspected function in the nucleus.
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  • 58
    ISSN: 1432-0878
    Keywords: Key words: Endometrium ; Epithelial cells ; Cell culture ; Polarization ; Steroid hormones ; Hormone receptors ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Uterine epithelial cells from normal human endometrium were cultured as a primary cell culture in a dual-chambered system. The epithelial cells were isolated from endometrial tissue of the proliferative phase obtained by hysterectomy. The epithelial cells were seeded on Millicell CM filters coated with the extracellular matrix Matrigel. Depending on the culture conditions, the epithelial cells formed a polarized cell monolayer on Matrigel or gland-like structures in Matrigel. The epithelial cell polarity was maintained during culture, which could be proved by electron microscopy. The progesterone and estrogen receptors as typical marker molecules for physiologically intact endometrial epithelial cells could be detected immunohistochemically as well as by RT-PCR in vitro and were down-regulated by medroxyprogesterone acetate (MPA) used as progesterone analogue. As this cell culture system exhibits morphological and immunohistochemical characteristics, typical for the in vivo situation, and since it can be modulated by hormone treatment under the in vitro conditions described, it represents a valuable tool for investigating processes that are essential for endometrial differentiation and reproductive functions.
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  • 59
    ISSN: 1432-0878
    Keywords: Key words Salivary gland ; Secretion ; Granule docking ; Exocytosis-endocytosis coupling ; OsO4 maceration ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The luminal membrane of salivary acinar cells creates a specialized cell surface area that accepts exocytosis and undergoes dynamic changes during secretion. These changes were visualized three-dimensionally from both the inside and outside of the cell in human parotid and submandibular glands, by application of in vitro secretory stimulation and then of OsO4 maceration to remove cytoplasmic organelles by varying degrees. In control glands treated without secretagogues, the luminal surface of serous acinar cells bore well-developed microvilli with only an occasional incidence of exocytotic profiles. Following treatment with the β-adrenergic agonist, isoproterenol, considerable shortening and loss of microvilli occurred along the luminal membrane where, on its cytoplasmic side, many protuberances of sizes similar to or smaller than those of single secretory granules (∼1 μm in diameter) appeared. The cytoplasmic surface of these protuberances exhibited small vesicles (∼100–150 nm in diameter) that, by transmission electron microscopy, were shown to be coated pits or vesicles present on or around the exocytosed granule membranes. Treatment of tissues with the muscarinic agonist carbachol also caused a decrease of microvilli and the appearance of protrusions at the luminal membrane. However, unlike isoproterenol treatment, many of these protrusions were devoid of small pits or vesicles and were much larger than a single secretory granule. These results indicate that (1) secretory stimulation causes the dynamic transformation of microvilli at the luminal membrane, where granule docking and membrane fusion take place, and (2) after fusion, the exocytosed membranes are processed differently, by coated pit/vesicle mediated or non-mediated mechanisms, according to the autonomic receptor control.
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  • 60
    ISSN: 1432-0878
    Keywords: Key words: Neurofilaments ; Phosphorylation ; Differentiation ; Immunocytochemistry ; Brain storage ; Fixation ; Microwave ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Neurofilaments, which are exclusively found in nerve cells, are one of the earliest recognizable features of the maturing nervous system. The differential distribution of neurofilament proteins in varying degrees of phosphorylation within a neuron provides the possibility of selectively demonstrating either somata and dendrites or axons. Non-phosphorylated neurofilaments typical of somata and dendrites can be visualized with the aid of monoclonal antibody SMI 311, whereas antibody SMI 312 is directed against highly phosphorylated axonal epitopes of neurofilaments. The maturation of neuronal types, the development of area-specific axonal networks, and the gradients of maturation can thus be demonstrated. Optimal immunostaining with SMI 311 and SMI 312 is achieved when specimens are fixed in a mixture of paraformaldehyde and picric acid for up to 3 days and sections are incubated free-floating. Neurons, with their dendritic domains immunostained by SMI 311 in a Golgi-like manner, can be completely visualized in relatively thick sections. The limitations of Golgi-preparations, such as glia-labeling, artifacts, and the staining of only a small non-representative percentage of existing neurons, are not apparent in SMI preparations, which additionally provide the possibility of selectively staining axonal networks. The results achieved in normal fetal brain provide the basis for studies of developmental disturbances.
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  • 61
    ISSN: 1432-0878
    Keywords: Key words mRNA ; Cancerous epithelium ; Autocrine growth regulation ; In situ hybridization ; Immunohistochemistry ; Western blotting ; Benign prostate hyperplasia ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Insulin-like growth factors (IGFs) are potent mitogens for a variety of cancer cells in vitro. A paracrine/autocrine role of IGF-II in the growth of breast and prostate cancer cells has been suggested. Information on cell-type-specific IGF-II expression in vivo in the breast and prostate is, however, limited. Thus, cell types expressing IGF-II mRNA and protein in tumors were identified by in situ hybridization and immunohistochemistry. Of 36 prostate, 17 breast, and 10 bladder cancers, and 9 paraganglioma tissues examined, IGF-II was expressed in more than 50% of prostate, breast, and bladder tumors, and in 100% of paraganglioma tumors. Expression levels of IGF-II were highest in the paraganglioma and bladder followed by prostate and breast tumors. In all the tumors expressing IGF-II, both mRNA and protein were localized to malignant cells, expression in the stroma being minimal. Since previous studies had indicated that an incompletely processed form of 15-kDa IGF-II exhibited higher mitogenic potency than the completely processed 7.5-kDa IGF-II form, the quantity and size of IGF-II proteins expressed in these tumors were analyzed by Western immunoblotting. Greater expression of 15-kDa IGF-II relative to the 7.5-kDa IGF-II form was clearly demonstrated in all six prostate cancers and in half of the two breast and four bladder cancers examined. The results are consistent with the hypothesis that the 15-kDa form of IGF-II expressed in cancerous cells contributes to autocrine cancer cell growth in vivo.
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  • 62
    ISSN: 1432-0878
    Keywords: Key words Interleukin-18 ; Hyperosmotic conditions ; Epithelial cells ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Interleukin-18 is a novel multifunctional cytokine, which enhances natural killer cell activity and promotes the induction of cytokine production, including that of interferon-γ by T cells and antitumor effects. Interleukin-18 is produced by cells of several different tissues (e.g., macrophages, keratinocytes, osteoblasts, and intestinal epithelium); however, it is unclear what physiological conditions or stimuli induce interleukin-18 production. To determine physiological conditions for the production of interleukin-18, we have examined the effect of mannitol-induced hyperosmotic conditions on normal human umbilical vein endothelial cells (HUVEC) and eight established human epithelial-like cell lines (Intestine 407, Caco-2, A253, HeLa, SCC25, HT1197, ACHN, A549). Hyperosmotic conditions induced interleukin-18 immunoreactivity in all the human cell lines tested, as detected by immunocytochemistry. The enhanced interleukin-18 production was also observed when mannitol was replaced with NaCl as the inducer of hyperosmotic stress. Enzyme-linked immunosorbent assays revealed that interleukin-18 concentrations in cell extracts were significantly increased by hyperosmotic conditions. Reporter gene assays also revealed that hyperosmotic conditions stimulated transcriptional activity of the interleukin-18 promoter. These results show for the first time that hyperosmotic stress is a stimulator of interleukin-18 production in epithelial-like cells.
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  • 63
    ISSN: 1432-0878
    Keywords: Key words Alkaline phosphatase ; Caco-2 intestinal epithelial cells ; Differentiation ; Dipeptidyl dipeptidase ; Proliferation ; Tyrosine kinase ; Tyrosine phosphoproteins ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Intestinal epithelial cell differentiation is closely regulated during normal cell renewal, maturation, and malignant transformation. Since tyrosine phosphorylation influences differentiation in other cell types and has been reported to vary between crypt cells to differentiated villus tip cells, we investigated the influence of tyrosine phosphorylation in colonocyte differentiation, by using human colonic Caco-2 cells as a model and expression of the brush border enzymes alkaline phosphatase (AKP) and dipeptidyl peptidase (DPDD) as differentiation markers. We studied three tyrosine kinase inhibitors with different modes of action and specificities, viz., genistein, erbstatin analog (EA), and tyrphostin, and the tyrosine phosphatase inhibitor sodium orthovanadate. AKP- and DPDD-specific activities were assayed in protein-matched cell lysates by synthetic substrate digestion. We also correlated the effects of these agents on brush border enzyme activity with tyrosine phosphorylation of phosphoproteins by Western blotting. Genistein (5–75 mg/ml) dose-dependently stimulated AKP and DPDD with a maximal stimulation at 75 mg/ml by 158.6± 17.5% and 228.6±37.1% of control values, respectively (n=12, P〈0.001). The inactive analog genistin had no effect. Tyrphostin (25 mM) similarly stimulated AKP and DPDD by 138.6±6.6% and 131.8±1.5% of control values (n=12, P〈0.001). Unexpectedly, EA (0.1–10 mM) had the opposite effect, inhibiting AKP- and DPDD-specific activity significantly at 10 mM with a maximal 14.8±6.4% and 26.5±2.5% of control values (n=12, each P〈0.001). Sodium orthovanadate had a discordant effect on these two differentiation markers. Orthovanadate dose-dependently increased AKP to a maximal 188.5±16.1% of basal activity at 1.5 mM but decreased DPDD activity at 1.5 mM to 47.2±3.8% (n=9, P〈0.001 each). The effects of each agent were preserved when proliferation was blocked with mitomycin C, suggesting that the modulation of phenotype by these agents was independent of any effects of proliferation. The tyrosine phosphorylation of several phosphoprotein bands was affected differently by these agents. In particular, the tyrosine phosphorylation of one 70-kDa to 71-kDa band was increased by genistein and tyrophostin but deceased by EA. The different effects of these modulators of tyrosine kinase activity raise the possibility that at least two independent enzymes or pathways regulating tyrosine phosphorylation modulate intestinal epithelial differentiation. Furthermore, tyrosine phosphorylation of the 70-kDa to 71-kDa phosphoprotein may be important in the intracellular signaling by which intestinal epithelial cell differentiation is controlled.
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  • 64
    ISSN: 1432-0878
    Keywords: Key words: Teeth ; Dental pulp ; Immunohistology ; Extracellular matrix ; Collagen ; Aging ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The distribution of extracellular matrix molecules, especially collagen types I, III, V, and VI, in the extracellular matrix of the connective tissue of human dental pulp of various ages was studied by polarization and indirect immunofluorescence microscopy by using a conventional fluorescence microscope and a confocal laser scanning microscope. Polarization and immunofluorescence microscopy of paraffin sections showed thick fibers of collagen type I, which represented the main component of the connective tissue matrix of the dental pulp. By indirect immunofluorescence, thin fibers and small bundles of collagen type III were determined to be one of the main fibrillar elements present in the dental pulp matrix. Collagen type IV was detected by a clear intense staining of the basement membrane of blood vessels at all ages examined. Collagens type V and VI formed a dense meshwork of thin microfibrils throughout the stroma of the connective tissue of the dental pulp. These fibers were localized around blood vessels and appeared to be enriched in the subodontoblastic layer. Investigations by means of confocal laser scanning microscopy revealed fibers of collagen type VI spiralling between fully differentiated odontoblasts toward the predentin layer. With advancing age, the connective tissue matrix appeared to be condensed and aggregates of thick fiber bundles could be observed. Furthermore, the participation of various collagen types in the composition of pulp stones was shown. These calcifications and diffuse calcifications increased in frequency with advancing age in a statistically significant manner.
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  • 65
    ISSN: 1432-0878
    Keywords: Key words: Small proteoglycans ; Intervertebral disc ; Decorin ; Biglycan ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Immunohistochemistry was used to study the presence and distribution of the core proteins of the small proteoglycans decorin and biglycan in the various compartments of human intervertebral discs. Both proteoglycans could be found in the outer tendon-like parts of the annulus fibrosus, indicating their potential role in collagen network formation and biomechanical stress resistance. The loss of both proteoglycans in the annulus of individuals older than 50 years reflects a normal age-related change. In the nucleus pulposus, decorin could be found in fibrillar areas of the interterritorial matrix, thereby indicating co-localization of decorin with fibrils containing type II collagen. Biglycan was present in the extracellular matrix of the nucleus pulposus of adults. The pericellular immunoreactive rims observed around nucleus pulposus cells and giant chondrones indicated local biosynthetic activity for these small proteoglycans. The staining patterns in cartilage endplates resembled those found in human hyaline articular cartilage.
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  • 66
    ISSN: 1432-0878
    Keywords: Key words: Collagen ; Matrilysin (PUMP) ; Wound healing ; Tumors ; Fat-storing cells ; Peripheral nerve glial cells ; Fibrocytes ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Matrix metalloproteinases represent a family of zinc-dependent proteolytic enzymes thought to be involved in normal and disease-related tissue remodeling processes. Increasing information about these enzymes is becoming available concerning their primary sequences, regulation at the mRNA level, activation of proenzymes, and modulation of enzyme activity by tissue inhibitors. In contrast, their morphological distribution and biological functions in normal tissues are poorly understood. In the present report, the comparative distribution of five members (gelatinase-A, gelatinase-B, matrilysin, stromelysin-1, and stromelysin-3) of the matrix metalloproteinase family and of one inhibitor (TIMP-1) has been morphologically analyzed in human liver and skin with the aid of new monospecific antibodies. Because of their common designation as matrix proteinases, these enzymes might have been expected to be distributed throughout these tissues, or at least in the connective tissue. However, each member of the family produces a highly specific pattern, staining structures such as arteriolar smooth muscle cells, myoepithelial cells in secretory portions or the luminal lining in excretory ducts of dermal sweat glands, liver bile canaliculi, or structures surrounding peripheral nerve axons. No reactivity is detected in rat tissues.
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  • 67
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    Cell & tissue research 289 (1997), S. 207-210 
    ISSN: 1432-0878
    Keywords: Key words: Parkinson’s disease ; Programmed cell death ; Dopamine ; Growth factors ; Neurotransplantation ; Glia ; Human ; Bonnet monkey ; Macaca radiata (Primates)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Dopamine cell death is the primary problem limiting the value of neurotransplantation in human patients with Parkinson’s disease. To address this problem, we added glial cell line-derived neurotrophic factor (GDNF) to cultures of embryonic dopaminergic neurons obtained from human and from Bonnet monkey (Macaca radiata) in an effort to reduce apoptotic cell death and improve overall cell survival. Tissue from three human embryos, 7–8 weeks post-conception, and one 9-week post-conception monkey embryo were dissociated and cultured in F-12 media with 5% human placental serum. GDNF (10 ng/ml) in human cultures nearly doubled dopamine neuron survival and reduced the rate of apoptosis from 6% to 3%. In monkey cultures, GDNF also enhanced dopamine neuron survival and reduced the apoptotic rate. We conclude that GDNF improves the survival of primate embryonic dopamine neurons in culture by reducing apoptosis.
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  • 68
    ISSN: 1432-0878
    Keywords: Key words: Complement C1s ; Sandwich ELISA ; Chondrocytes ; Cell culture ; Differentiation ; Ascorbic acid ; Syrian hamster ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. In vitro synthesis of the first component of complement C1s was examined by using hamster epiphyseal chondrocytes (HAC) and human chondrosarcoma cell line HCS-2/8. Hamster and human C1s produced by the cells were quantified by immunoblotting and sandwich enzyme-linked immunosorbent assay (ELISA), respectively. It was possible to measure active and inactive C1s by sandwich ELISA, when we used anti-human C1s monoclonal antibodies, M241 recognizing only active C1s, and M365 and M81 recognizing both active and inactive C1s. Approximately 40% of C1s secreted from HCS-2/8 was found to be activated in the culture medium, whereas C1s from HAC was not. C1s production increased in accordance with chondrocyte differentiation induced by ascorbic acid. In contrast, transforming growth factor-beta1 and basic fibroblast growth factor, which inhibited differentiation, suppressed C1s production. These results confirmed our previous observation showing that C1s synthesis increased with differentiation into hypertrophic chondrocytes in vivo.
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  • 69
    ISSN: 1432-0878
    Keywords: Key words Vinculin ; Talin ; Integrin ; Dystrophin ; Spectrin ; T-tubule ; Costamere ; Basal membrane ; Cardiac muscle cell ; Dilated cardiomyopathy ; Human ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The transverse tubule system of the cardiomyocyte remains undeformed despite the extreme forces it undergoes during the contraction-relaxation cycle, but the morphological basis for its stability remains unclear. Therefore, we have investigated the architecture and subcellular protein scaffold of the cardiac T-tubules and compared it with that of the costameres and of the free sarcolemma. Tissue samples from normal rat and monkey hearts, and left ventricular tissue from normal and cardiomyopathic human hearts obtained at transplantation surgery were investigated using immunocytochemistry and confocal microscopy and by electron microscopy. In addition, we used a re-differentiation model of isolated, cultured adult rat cardiomyocytes. The cell membrane of the cardiac T-tubules was found to contain the cell-matrix focal adhesion molecules (FAMs) vinculin, talin, the α5β1 integrin and the membrane-associated proteins (MAPs) dystrophin and spectrin. FAMs and MAPs were localized in the T-tubular membrane in a similar pattern: in longitudinally oriented myocytes as transverse punctate lines at the Z-level; in transversally cut myocytes a radial tubular network was found to extend throughout the interior of the cell. Immunolabeling for basement membrane components including collagen IV, fibronectin and laminin showed a colocalization with FAMs and MAPs parallel to the transverse T-tubules. The costameres of the sarcolemma showed a protein composition resembling that of the T-tubules but the intervening segments of free sarcolemma showed absence of FAMs and presence of MAPs. For the first time, we demonstrate the existence and protein composition of the T-tubular scaffold in the human heart. Furthermore, we show that cardiomyocytes from human failing hearts have less abundant but more dilated T-tubules than do experimental animals. These results indicate that the cardiac T-tubular system contains a subcellular scaffold closely resembling that of the costameres. It consists of FAMs, MAPs and basal lamina proteins that confer structural integrity to the cardiac T-tubular membrane during contraction/relaxation cycles.
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  • 70
    ISSN: 1432-0878
    Keywords: Key words α1-Microglobulin ; Bikunin ; Inter-α-trypsin inhibitor ; Mast cells ; Trypstatin ; Urinary trypsin inhibitor ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  We recently reported that the rat mast cell proteinase inhibitor trypstatin is genetically identical with the second half of inter-α-trypsin inhibitor light chain (ITI-LC), also known as bikunin or urinary trypsin inhibitor (UTI). In this study, therefore, immunoreactivities of mast cells of various human tissues were examined with three antibodies, anti-human ITI-LC, anti-ITI, which recognizes mainly heavy chains or the sugar moiety of ITI, and anti-α 1-microglobulin (α1mG). ITI-LC immunoreactivity was strongly found in mast cells in the connective tissues of various organs except for those of the propria mucosae of small intestine. Neither anti-ITI antibody nor anti-α1mG antibody reacted with mast cells in various tissues. By reverse transcription-polymerase chain reaction (RT-PCR) analysis, α1mG/ITI-LC mRNA was not detected in the skin and tongue, and only weakly in small intestine, although ITI-LC immunoreactivity was strongly detected in these tissues. Furthermore, the mRNA was not expressed in cultured human mast cells. These results suggest that ITI-LC protein is stored in the granules of human connective tissue mast cells, though is not produced by them.
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  • 71
    ISSN: 1432-0878
    Keywords: Key words Osteoblasts ; Extracellular matrix ; Transforming growth factor-β1 ; Basic fibroblast growth factor ; Apert’s syndrome ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The phenotype of Apert osteoblasts differs from that of normal osteoblasts in the accumulation of macromolecules in the extracellular matrix. Apert osteoblasts increase type I collagen, fibronectin and glycosaminoglycans secretion compared with normal osteoblasts. Because the extracellular matrix macromolecule accumulation is greatly modulated by transforming growth factor-β1, we examined the ability of normal and Apert osteoblasts to secrete transforming growth factor-β1 by CCL-64 assay and to produce transforming growth factor-β1 by analysis of the mRNA expression of transforming growth factor-β1. Northern blot analysis revealed an increased amount of transforming growth factor-β1 mRNA expression in Apert osteoblasts compared with normal ones. Moreover, the level of the active transforming growth factor-β1 isoform was higher in Apert than in normal media. In pathologic cells, the increase in transforming growth factor-β1 gene expression was associated with a parallel increase in the factor secreted into the medium. The level of transforming growth factor-β1 was decreased by the addition of basic fibroblast growth factor. Transforming growth factor-β1 is controlled temporally and spatially during skeletal tissue development and produces complex stimulatory and inhibitory changes in osteoblast functions. We hypothesise that in vitro differences between normal and Apert osteoblasts may be correlated to different transforming growth factor-β1 cascade patterns, probably due to an altered balance between transforming growth factor-β1 and basic fibroblast growth factor.
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  • 72
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    Cell & tissue research 290 (1997), S. 39-42 
    ISSN: 1432-0878
    Keywords: Key words: Ciliogenesis ; Striated rootlets ; Oviduct ; Ciliated cells ; Ultrastructure ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Striated rootlets in ciliated cells are conical banded structures composed of longitudinally aligned filaments. The formation of striated rootlets during ciliognesis in the human oviduct epithelium was studied by electron microscopy. Primitive rootlets appeared at the proximal side of basal bodies before or at the same time as ciliary budding. After the formation of several striations, the tip of the rootlets extended deeply toward the interior of the cell and became differentiated into two distinct parts, viz., the proximal conical part connected to the basal body and the distal fibrillar part. The periodicity of the striations in the fibrillar part was 68.5±2.95 nm, about 5 nm longer than that of the conical part (63.9±2.25 nm). The dark band in the striation was thicker in the fibrillar part than in the conical part. Since the fibrillar part was not observed in the mature cilium, this part was considered as being either degraded or changed into the conical part during ciliogenesis.
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  • 73
    ISSN: 1432-0878
    Keywords: Key words Prion protein (PrPc) ; Electron microscopy ; Secretory granules ; Membrane ; Extracerebral tissues ; Hamster ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  In transmissible spongiform encephalopathies (TSE), such as scrapie in animals and Creutzfeldt-Jakob disease in humans, the central event is the conversion of a host-encoded amyloidogenic protein (PrPc) into an abnormal isoform (PrPsc) that accumulates as amyloid in TSE brain. PrPc is a membrane sialoglycoprotein synthesized in the central nervous system and elsewhere. We have examined the ultrastructural localization of PrPc in numerous hamster and some human extracerebral tissues, by means of a post-embedding electron-microscopic method combined with immunogold labeling. In stomach, intestine, lung, and kidney from hamsters, and in stomach, kidney, and spleen from humans, immunogold labeling specific for PrPc is observed on various cellular substructures related to secretory pathways: Golgi apparatus, secretory globules, and plasma membrane. In mucous epithelial cells of stomach and intestine, PrPc appears to be concentrated in secretory globules, suggesting a role for PrPc in the secretory function of the digestive tract. The secretory aspect of PrPc may be a key to understanding the physiopathological mechanisms underlying TSE.
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  • 74
    ISSN: 1432-0878
    Keywords: Key words Neurons ; Brain ; Chemotaxis ; Endothelial cells ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The formylpeptide receptor (FPR), previously found only on polymorphonuclear leukocytes and monocytes/macrophages, responds to both synthetic N-formyl oligopeptides and those produced by bacteria. The cDNA for human FPR has been cloned and a rabbit polyclonal antiserum directed against a synthetic 11-amino-acid peptide corresponding to the deduced carboxy-terminus has been produced. We have now extensively characterized and used the antibody to detect FPR on normal human tissues and cell types. The receptor antigen is present on some epithelial cells, especially those with a secretory function, and on some endocrine cells, e.g., follicular cells of the thyroid and cortical cells of the adrenal. Liver hepatocytes and Kupffer cells are positive. Smooth muscle and endothelial cells are also generally positive. In the brain and spinal cord, the neurons of the motor, sensory, and cerebellar systems, and those of the parasympathetic and sympathetic systems stain positively. These data suggest that the putative endogenous agonist for FPR or an antigenically similar receptor reacts with cellular targets in the neuromuscular, vascular, endocrine, and immune systems.
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  • 75
    ISSN: 1432-0878
    Keywords: Key words Organotypic cocultures ; Histodifferentiation ; Proliferation ; Basement membrane components ; Integrins ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Cell-matrix interactions and the ordered deposition of basement membrane (BM) components are of major importance for the maintenance of tissue homeostasis in complex epithelia. This aspect was studied in vitro in a coculture system designed as an oral mucosa model. As crucial epithelial features the kinetics of proliferation, expression of site-specific keratins as well as integrin patterns in correlation to synthesis of BM components were assessed by immunohistochemistry and in situ hybridization. Comparison with non-cornified gingiva as tissue of origin revealed different stages of epithelial development, eventually leading to complete reconstruction within a time frame of 1–3 weeks. First, the initial activated stage up to 1 week was characterized by (a) high keratinocyte proliferation, (b) extended expression of the basal cell-specific keratin K5 and (c) a patchy pattern of the differentiation-specific keratins K4 and K13. Second, after 2 weeks the improvement of histoarchitecture correlated to (a) predominant K5 expression in the basal and (b) extension of K4 and K13 within the suprabasal cell compartment, (c) high expression of integrins α3β1 and α6β4 including their ligand laminin-5 and (d) accumulating deposition of basement membrane components. Third, virtually complete tissue normalization at 3 weeks was indicated by (a) restriction of K5 to the basal cell area, (b) regular suprabasal localization of K4 and K13, (c) polarization of integrins to basal and parabasal cells and (d) linear codistribution of collagen IV, “classical” laminin (-1 or -10) and laminin-5 underneath the basal cells. Thus, these organotypic cocultures represent relevant equivalents for non-keratinized oral mucosa with typical gingival differentiation features and in addition suitable models for preclinical trials such as prospective dental material testing.
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  • 76
    ISSN: 1432-0878
    Keywords: Key words Endothelin ; Endothelin receptors ; Endothelin-converting enzyme ; Ovary ; In situ hybridization ; Human ; Cynomolgus monkey
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The endothelin system is composed of three endothelin isoforms (ET-1, ET-2, and ET-3), the endothelin receptors ETA and ETB, and the endothelin-converting enzyme (ECE). Besides having a major vasoactive role, endothelins have roles in different cell types at a local level. We investigated the presence of the different components of the endothelin system in primate ovaries. Human ovaries and gonadotropin-stimulated monkey ovaries were studied using immunohistochemistry for endothelin, and in situ hybridization with probes for ET-1, ET-2, ET-3, ETA and ETB receptors, and ECE. ET-1 and ETA receptors were detected in endothelial cells and vascular smooth muscle cells, respectively, in stromal vessels adjacent to follicles and corpora lutea. ETB receptors and ET-1 were found in the endothelial cells of capillaries of corpora lutea. ECE was present in internal theca cells of secondary, de Graaf, atretic follicles, and in luteinized granulosa cells of the corpora lutea. The endothelin system components are present in or around the follicles of human and monkey ovaries. Although the components are not expressed in the same cell types, they are synthesized, mainly in follicles, by cells that are in close proximity. Thus, the endothelin system could act in a paracrine manner. ECE expression in steroid-producing cells changes its compartmentalization during follicle maturation.
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  • 77
    ISSN: 1432-0878
    Keywords: Osteoblasts ; Preosteoclasts ; Cell differentiation ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Osteoblasts are involved in the bone resorption process by regulating osteoclast maturation and activity. In order to elucidate the mechanisms underlying osteoblast/preosteoclast cell interactions, we developed an in vitro model of co-cultured human clonal cell lines of osteoclast precursors (FLG 29.1) and osteoblastic cells (Saos-2), and evaluated the migratory, adhesive, cytochemical, morphological, and biochemical properties of the co-cultured cells. In Boyden chemotactic chambers, FLG 29.1 cells exhibited a marked migratory response toward the Saos-2 cells. Moreover, they preferentially adhered to the osteoblastic monolayer. Direct co-culture of the two cell types induced: (1) positive staining for tartrate-resistant acid phosphatase in FLG 29.1 cells; (2) a decrease of the alkaline phosphatase activity expressed by Saos-2 cells; (3) the appearance of typical ultrastructural features of mature osteoclasts in FLG 29.1 cells; (4) the release into the culture medium of granulocyte-macrophage colony stimulating factor. The addition of parathyroid hormone to the co-culture further potentiated the differentiation of the preosteoclasts, the cells tending to fuse into large multinucleated elements. These in vitro interactions between osteoblasts and osteoclast precursors offer a new model for studying the mechanisms that control osteoclastogenesis in bone tissue.
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  • 78
    ISSN: 1432-0878
    Keywords: Key words: Spleen ; Fetus ; Development ; Extracellular matrix ; Immuno-electron microscopy ; Transmission electron microscopy ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The distribution of the extracellular matrix proteins types III pN-collagen and IV collagen, laminin and tenascin was investigated in fetal, infant, and adult human spleens by using immuno-electron microscopy. The presence of type III pN-collagen was assessed by using an antibody against the aminoterminal propeptide of type III procollagen. All the proteins other than type III pN-collagen were found in reticular fibers throughout development. In the white pulp of the fetus aged 16 gestational weeks, only an occasional type III pN-collagen-containing fibril was present, although type III pN-collagen was abundant in the reticular fibers of the red pulp. Conversely, in adults, most of the reticular fibers of the white pulp, but not of the red pulp, were immunoreactive for type III pN-collagen. Ring fibers, the basement membranes of venous sinuses, were well developed in both infant and adult spleens. The first signs of their formation could be seen as a discontinuous basement membrane, which was immunoreactive for type IV collagen, laminin, and tenascin in the fetus aged 20 gestational weeks. Intracytoplasmic immunoreactivity for all the proteins studied was visible in the mesenchymal cells of the fetus aged 16 gestational weeks and in the reticular cells of the older fetuses, which also showed labeling for type IV collagen and laminin in the endothelial cells. The results suggest that proteins of the extracellular matrix are produced by these stationary cells.
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  • 79
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    Cell & tissue research 285 (1996), S. 171-176 
    ISSN: 1432-0878
    Keywords: Key words: Keratin ; Prostate ; Epithelium ; Differentiation ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Keratin 19 is found primarily in simple epithelia. In mammary epithelia, keratin 19 was localized to a subset of luminal cells, suggesting that keratin 19-negative cells may be the proliferative compartment of the secretory cell lineage. The structural and functional similarities of prostate and breast led us to examine keratin 19 expression in the prostate. Immunohistochemical studies revealed that keratin 19 expression was heterogeneous and frequently occurred in basal as well as in luminal cells of normal, dysplastic, and benign hyperplastic tissues. Keratin 19 was observed in cancer, but usually in a minority of cells. This was in dramatic contrast to invasive breast cancers, which are reportedly uniformly positive for keratin 19. Prostatic epithelial cells cultured from tissues of all histologies expressed keratin 19. In summary, keratin 19 does not obviously correlate with any epithelial cell lineage or phenotype in the prostate.
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  • 80
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    Cell & tissue research 280 (1995), S. 183-188 
    ISSN: 1432-0878
    Keywords: Blood platelets ; Oxidized LDL ; Cytochalasin D ; Reflection contrast microscopy ; Image analysis ; Electron microscopy ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The adhesion of human blood platelets is studied with an in vitro model using reflection contrast microscopy and an image analysis system. The adhesive feature is promoted by oxidatively modified low density lipoprotein, which also induces functional morphological changes of platelets. However, when washed platelets are pretreated with 0.05 mM cytochalasin D, oxidized low density lipoprotein (100 μg/ml) causes a slower increase of the adhesion area (11.6 μm2/min) compared to untreated platelets (15.7 μm2/min) or platelets treated by oxidized low density lipoprotein alone (20.5 μm2/min, P〈0.01). These results are supported by light transmission analysis and by transmission electron microscopy. Our experiments suggest that cytochalasin D inhibits the change of platelets in shape induced by oxidized low density lipoprotein, hinders the adhesion, but does not prevent the adhesion entirely.
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  • 81
    ISSN: 1432-0878
    Keywords: Key words: Galectin ; β-Galactoside-binding lectin ; Human ; Skin ; Immunocytochemistry ; Immunohistochemistry ; Hybridization ; in situ ; Langerhans cell ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The localization of an endogenous 14-kDa β-galactoside-binding lectin (galectin) and its pattern of gene expression were examined in normal human skin by light- and electron microscopy. Under the light microscope, immunostaining of 14-kDa galectin was observed in the cell membrane of cells in the basal and spinous layers of the epidermis. Galectin was also found in the Langerhans cells, as shown by double labeling using anti-14-kDa galectin and anti-CD1a antibodi es. In the dermis, immunostaining for the 14-kDa galectin was positive in the extracellular matrix and fibroblasts. At the electron-microscopic level of resolution, galectin was located primarily along the plasma membrane of keratinocytes, and in both the cytoplasm and nucleus of Langerhans cells in the epidermis, whereas in the dermis it was detected in the extracellular matrix and in both the nucleus and cytoplasm of fibroblasts. The gene expression of 14-kDa galectin was visualized by the HRP-staining me thod following in situ hybridization techniques. The expression was detected in the cytoplasm of cells in the basal and spinous layers of the epidermis; whereas, in the dermis, it was detected in the cytoplasm of fibroblasts. Moreover, SDS-polyacrylamide gel electrophoresis and lectin-blot analysis revealed that this galectin bound to glycoproteins of approximately 17, 62, and 72 kDa in the epidermis and to those of 29, 54, and 220 kDa in the dermis. The present study indicates that 1) normal human skin produces the β-galactoside-binding 14-kDa galectin, and 2) this galectin is located in both the epidermis, particularly in the keratinocytes and Langerhans cells, and in the dermis. These results suggest that galectin is important for cell-cell contact and/or adhesion in the epidermis and for cell-extracellular matrix interaction in the dermis.
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  • 82
    ISSN: 1432-0878
    Keywords: Galectin ; β-Galactoside-binding lectin ; Human ; Skin ; Immunocytochemistry ; Immunohistochemistry ; Hybridization, in situ ; Langerhans cell ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The localization of an endogenous 14-kDa β-galactoside-binding lectin (galectin) and its pattern of gene expression were examined in normal human skin by light- and electron microscopy. Under the light microscope, immunostaining of 14-kDa galectin was observed in the cell membrane of cells in the basal and spinous layers of the epidermis. Galectin was also found in the Langerhans cells, as shown by double labeling using anti-14-kDa galectin and anti-CD1a antibodies. In the dermis, immunostaining for the 14-kDa galectin was positive in the extracellular matrix and fibroblasts. At the electron-microscopic level of resolution, galectin was located primarily along the plasma membrane of keratinocytes, and in both the cytoplasm and nucleus of Langerhans cells in the epidermis, whereas in the dermis it was detected in the extracellular matrix and in both the nucleus and cytoplasm of fibroblasts. The gene expression of 14-kDa galectin was visualized by the HRP-staining method following in situ hybridization techniques. The expression was detected in the cytoplasm of cells in the basal and spinous layers of the epidermis; whereas, in the dermis, it was detected in the cytoplasm of fibroblasts. Moreover, SDS-polyacrylamide gel electrophoresis and lectin-blot analysis revealed that this galectin bound to glycoproteins of approximately 17, 62, and 72 kDa in the epidermis and to those of 29, 54, and 220 kDa in the dermis. The present study indicates that 1) normal human skin produces the β-galactoside-binding 14-kDa galectin, and 2) this galectin is located in both the epidermis, particularly in the keratinocytes and Langerhans cells, and in the dermis. These results suggest that galectin is important for cell-cell contact and/or adhesion in the epidermis and for cell-extracellular matrix interaction in the dermis.
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  • 83
    ISSN: 1432-0878
    Keywords: Key words: SS-B/La protein ; La cDNA ; Autoimmunity ; In situ hybridization ; Gene expression ; Endothelium ; Blood vessels ; Liver ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The gene for the nuclear autoantigen La/SS-B encodes two La mRNA isoforms. In order to study the function and expression of both La mRNA forms, an in situ hybridization procedure was developed allowing the selective identification of either exon 1 or exon 1′. For this purpose, digoxigenin-labeled exon-specific sense and anti-sense probes were prepared by in vitro transcription from plasmids that contained the respective exon sequence. Detection of the probes was carried out by using rhodamine-conjugated anti-digoxigenin antibody and confocal laser scanning microscopy. Both La mRNAs were found in the cytoplasm of endothelial cells but not in smooth muscle cells. In addition to the in situ technique, an assay system was established allowing the expression ratio of the two mRNA forms to be determined. The estimation was based on the amplification of exon 1 and 1′ La cDNAs in parallel by using a three primer polymerase chain reaction. The ratio of the exon 1 to exon 1′ La mRNA forms was determined to be about 5:1 in liver tissue and endothelial cells. The data support the conclusion that both La mRNA forms represent finally processed cytoplasmic mRNAs that are up- or downregulated in parallel.
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  • 84
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    Cell & tissue research 290 (1997), S. 623-631 
    ISSN: 1432-0878
    Keywords: Key words: PECAM-1 (platelet/endothelial cell adhesion molecule-1) ; Endothelium ; HUVEC (human umbilical vein endothelial cells) ; Myocardium ; Ultrastructure ; Human ; Rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The subcellular localization of PECAM-1 in endothelial cells was examined by using advanced morphological techniques, such as confocal scanning microscopy and immunolabeling procedures for electron microscopy. The localization of PECAM-1 was studied immunohistochemically with five specific monoclonal antibodies and one polyclonal antibody (all anti-human) in human and rabbit myocardium and in isolated endothelial cells. In vivo, PECAM-1 was localized uniformly on the plasma membrane of all vascular endothelial cells, predominantly on the luminal side of vessels. No specific increase in labeling was found at sites of cell-to-cell contact. In vitro, primary isolated cells (human umbilical vein endothelial cells) showed continuous labeling of the entire cell membrane. Cells of higher passages were labeled in a manner similar to freshly isolated cells. Our findings refute the commonly accepted hypothesis that PECAM-1 is localized only at cell-to-cell contacts. Further, we have not been able to confirm the hypothesis regarding the important mechanical role of PECAM-1 in stabilizing the endothelial monolayer. Since PECAM-1 is also expressed on platelets and is known to bind to itself, the way in which PECAM-1-positive endothelial cells are protected against binding of PECAM-1-positive platelets remains unclear. In view of these findings, the role of PECAM-1 in the leukocyte migration cascade needs to be re-evaluated.
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  • 85
    ISSN: 1432-0878
    Keywords: Key words: Quinolinic acid ; Interferon-γ ; Kynurenine ; Electron microscopy ; Immunocytochemistry ; Excitotoxicity ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract.  Quinolinate (QUIN), a tryptophan-derived excitotoxin, was localized ultrastructurally in human peripheral blood monocytes/macrophages (MØ) by immuno-electron microscopy. A combined carbodiimide/glutaraldehyde/paraformaldehyde-based fixation procedure was developed for optimal retention of QUIN in the cell as well as minimal loss of ultrastructure; a silver-enhanced colloidal gold detection system was used for electron-microscopic analysis. Gold particles representing QUIN immunoreactivity were associated with the inner side of the plasma membrane in normal MØ. The number of gold particles increased significantly when QUIN levels were elevated by treatment with its precursor kynurenine, but location of the gold particles remained essentially the same under this condition. Treatment with interferon-γ increased the number of Golgi bodies, vacuoles and pseudopodia, reflecting the activated state of the cell. Significantly increased numbers of gold particles representing QUIN were detectable in approximately the same location as in the case of kynurenine treatment. Combined treatment with kynurenine and interferon-γ maximally increased the number of gold particles at the periphery of the cell. The pseudopodia were intensely stained with gold particles, while they were not detectable in the inner part of the cytoplasm or in any other organelle even under this activated condition. The significance of the specific location of QUIN revealed in the present study and its relation to the release and subsequent actions of QUIN are discussed.
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  • 86
    ISSN: 1432-0878
    Keywords: Key words Placenta ; Extravillous trophoblast ; Matrix-type fibrinoid ; Extracellular matrix ; MMP ; TIMP ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The invasion of extravillous trophoblast cells into the maternal endometrium is one of the key events in human placentation. The ability of these cells to infiltrate the uterine wall and to anchor the placenta to it as well as their ability to infiltrate and to adjust utero-placental vessels to pregnancy depends, among other things, on their ability to secrete enzymes that degrade the extracellular matrix. Most of the latter enzymes belong to the family of matrix metalloproteinases. Their activity is regulated by the tissue inhibitors of matrix metalloproteinases. We have studied the distribution patterns of matrix metalloproteinases-1, -2, -3, and -9 and their inhibitors TIMP-1 and TIMP-2 as compared to the distribution of their substrates along the invasive pathway of extravillous trophoblast of 1st, 2nd, and 3rd trimester placentas by means of light microscopy on paraffin and cryostat sections as well as at the ultrastructural level (only 3rd trimester placenta). The comparison of different methods proved to be necessary, since the immunohistochemical distribution patterns of these soluble enzymes are considerably influenced by the pretreatment of tissues. All three methods revealed immunoreactivities of both, proteinases and their inhibitors, not only intracellularly in the extravillous trophoblast but also extracellularly in its surrounding matrix, the distribution patterns depending on the stage of pregnancy and on the degree of differentiation of trophoblast cells along their invasive pathway. Within the extracellular matrix, immunolocalization of matrix metalloproteinases as well as their inhibitors showed a specific relation to certain extracellular matrix molecules.
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  • 87
    ISSN: 1432-0878
    Keywords: Key words: NO/cGMP pathway ; Testis ; Leydig cells ; Immunocytochemistry ; RIA ; Cell culture ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. In this study we sought to determine whether the main components of the nitric oxide (NO) pathway are localized within the Leydig cells of the human testis and whether the soluble guanylyl cyclase (sGC), the enzyme that accounts for NO effects, is functionally active in these cells. Using an amplified immunocytochemical technique, immunoreactivity for nitric oxide synthase (NOS-I), sGC and cyclic guanosine monophosphate (cGMP) was detected within the cytoplasm of human Leydig cells. Distinct differences in staining intensity were found between individual Leydig cells, between cell groups and between Leydig cells of different patients. By means of a specific cGMP-RIA, a concentration-dependent increase in the quantity of cGMP was measured in primary cultures of human Leydig cells following exposure to the NO donor sodium nitroprusside. In addition, NOS-I immunoreactivity was seen in Sertoli cells, whereas cGMP and sGC immunoreactivity was found in Sertoli cells, some apically situated spermatids and residual bodies of seminiferous tubules. Dual-labelling studies and the staining of consecutive sections showed that there are several populations of Leydig cells in the human testis. Most cells were immunoreactive for NOS-I, sGC and cGMP, but smaller numbers of cells were unlabelled by any of the antibodies used, or labelled for NOS-I or cGMP alone, for sGC and cGMP, or for NOS-I and sGC. These results show that the Leydig cells possess both the enzyme by which NO is produced and the active enzyme which mediates the NO effects. There are different Leydig cell populations that probably reflect variations in their functional (steroidogenic) activity.
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  • 88
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    Cell & tissue research 295 (1999), S. 297-305 
    ISSN: 1432-0878
    Keywords: Key words Trophoblast ; Macrophages ; Placenta ; Cell culture ; Paracrine regulation ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract In view of the accumulating evidence for paracrine mechanisms regulating trophoblast function, we tested the hypothesis that placental macrophages affect trophoblast activity in a paracrine fashion. Trophoblast was isolated from 17 term placentas (–IP). One aliquot of cells was further immunopurified (+IP) using an HLA class I antibody. This increased the proportion of trophoblast (+IP 〉97%; –IP ∼70%) as identified by rigorous immunocytochemistry. Most (∼70%) non-trophoblast cells in –IP were macrophages. The cells were cultured for 5 days with a daily medium change. In addition, +IP cells from seven placentas were cultured with lipopolysaccharide (LPS)-stimulated or -unstimulated macrophage-conditioned media. The concentrations of lactate, trophoblast-specific hormones, human chorionic gonadotropin-β (hCG-β) and human placental lactogen (hPL), of several prostanoids and of endothelin-1 and angiotensin II were determined in the culture media. The accumulated amounts of substances released into the culture media, corrected for the greater proportion of trophoblast in +IP cultures, were on average two- to threefold higher (hCG-β: 18-fold) in +IP than in –IP, with the exception of endothelin-1,2 (no change), angiotensin II (–70%) and 6-keto-prostaglandin-F1α (–40%). [3H]leucine incorporation into the trichloroacetic acid (TCA)-precipitable pool measured on day 5 was twofold higher in +IP than in –IP. Addition of conditioned media reverted these changes. The data demonstrate that placental macrophages in culture affect trophoblast biosynthetic activity in a paracrine fashion. We conclude that macrophages are important regulators of trophoblast activity.
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  • 89
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    Cell & tissue research 287 (1997), S. 335-342 
    ISSN: 1432-0878
    Keywords: Key words: Lysosomal membrane antigen ; Immunohistochemistry ; Biosynthesis ; Prostate-membrane-specific antigen ; Apocrine secretion ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The tissue distribution, preferentially in the human male genital system, and the subcellular localization of the lysosome-associated membrane protein 2 (lamp 2) was studied immunohistochemically using a mouse monoclonal antibody, 2D5. Strong immunoreactivity was present in the tubular system of the kidney, in acinar cells of salivary glands and pancreas, prostate, mammary glands, placenta and in cutaneous sweat glands. Moderate immunoreactivity was observed in cerebral neuronal cells, epidermal cells, testis, epididymis, seminal vesicle and endometrium. Very low immunoreactivity was found in liver. In some of the tissues mentioned, the distribution pattern of immunoreactivity is smooth and homogeneous, while in others it is granular and concentrated in the supra- or perinuclear cytoplasm. The subcellular distribution was studied on ultracryosections and on pre-embedding-processed chopper sections of human prostate. In the latter gland, the protein is not restricted to epithelium, but is also present in stromal cells. Ultrastructurally, the immunoreactivity in secretory cells was localized in electron-translucent vacuoles and granules, including the secretory granules. A close association with cell membranes was not generally the case. Only part of the immunoreactive material was linked to the apical plasma membrane pointing to a biosynthesis independent from an association step with the apical plasma membrane. As shown by immunoelectron microscopy and Western blotting, a high amount of lamp 2 is secreted and is found in so-called prostasomes. The findings indicate that in the human prostate most of the membrane-bound lamp 2 is released from the secretory cells, presumably in an apocrine fashion.
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  • 90
    ISSN: 1432-0878
    Keywords: Key words: Urothelium ; Tartrate-resistant acid phosphatase ; Nitric oxide synthase I ; Superoxide dismutase ; Immunocytochemistry ; Free radicals ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Three enzymes, viz., tartrate-resistant acid phosphatase (TRAP), nitric oxide synthase I (NOS-I), and superoxide dismutase (SOD), involved in the production and metabolism of free radicals or radical equivalents, were demonstrated by immunocytochemistry in the urothelium of the ureters of six patients of various ages. Two of these enzymes (TRAP and NOS-I) were colocalized in the most apical and lateral border of the superficial cells of the urothelium. In contrast, SOD showed a patchy or granular distribution within the supranuclear region of these cells. Intra- and subepithelial macrophages exhibited a weak TRAP, but no NOS-I or SOD, immune reaction. On the basis of the immunocytochemical findings, arguments in favor of a cytotoxic function of the superficial cells of the human urothelium are presented.
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  • 91
    ISSN: 1432-0878
    Keywords: Key words Airways ; Mucins ; Secretion ; Mini-organ culture ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  To study the secretory products and the proliferation of cells of the human respiratory surface epithelium, we established a miniorgan-culture system of bronchial tissue. Biopsies of large airways were grown on agar-coated dishes immersed in a serum-enriched medium. As determined by light and transmission electron microscopy, between 1 and 3 weeks, the organ cultures were covered by a differentiated epithelium consisting of secretory, ciliated, and basal cells. Immunohistochemistry, using antibodies to mucin and lysozyme, and lectin histochemistry revealed both mucous and serous secretory cells in the epithelium. Cell proliferation was studied in situ using antibodies to proliferating cell nuclear antigen (PCNA) and Ki-67. Whereas at the time of explantation the proliferation was low (2.5±1.7% of the epithelial cells were PCNA-positive, 1.7±0.6 were Ki-67-positive), at 24 h of cultivation, 30.4±5.1% or 25.2±4.9% of the epithelial cells were labeled with antibodies to PCNA or Ki-67. After 7 days, the number of dividing cells was low again. The results show that the organ-culture system of human respiratory surface epithelium produces a differentiated epithelium that is useful in the study of secretory processes, differentiation, and proliferation.
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  • 92
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    Cell & tissue research 290 (1997), S. 31-37 
    ISSN: 1432-0878
    Keywords: Key words: Sural nerve ; Blood vessels ; Angiopathic neuropathy ; Vasculitis ; Peripheral neuropathy ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The number and dimensions of epineurial blood vessels in normal human sural nerves have, thus far, not been determined using systematic, reproducible morphometric methods, although this nerve is most frequently used for diagnostic biopsies. Quantitative changes in epineurial blood vessels appear to be major parameters for identifying angiopathy and angiopathic peripheral neuropathy. Therefore, we examined the epineurial blood-vessel number in relation to the age of the patients and to the number and size of the nerve fascicles in each of 51 human sural nerve biopsies. The data from a control group were compared with pathological cases. We found that the number of epineurial blood vessels (normal mean: 57.7) increased significantly (up to 196) in biopsies where there were signs of angiopathy (P≤0.01) or vasculitis (P≤0.05). The increase in the number of epineurial blood vessels usually resulted from a proliferation of capillaries. The fascicular cross-sectional area did not appear to be related to the number of epineurial blood vessels, although it increased significantly in cases with vasculitis (P≤0.05) or an axonal type of neuropathy (P≤0.05). Thus, this study shows that the number of epineurial blood vessels is a helpful parameter in verifying angiopathy and angiopathic peripheral neuropathy.
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  • 93
    ISSN: 1432-0878
    Keywords: Key words: Serum-free culture medium ; Organ culture ; Fetal digits ; Fetal skin ; Nails ; Hair follicles ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. We have optimized a suspension organ culture (SOC) system previously used to study human fetal skin in vitro. Eleven types of media were used to culture fetal digits and skin samples (ranging from 59–91 days of estimated gestational age) for periods of 3 or 4 weeks. The cultures were supplemented with 0%–10% bovine calf serum and exposed to 5%–45% oxygen atmospheres. Optimal conditions, which were selected based upon the morphologic and histologic properties of the cultured tissues, were: DMEM-F12 medium in a serum-free environment with a 45% oxygen atmosphere. The refined SOC system supports the maintenance and/or continued development of the epidermis and dermis of trunk, back, scalp, and digital skin, developing hair follicles, nails, and glands. Overall, development and differentiation mimicked the in vivo state more closely than that observed when using other in vitro culture systems. The effects of bovine calf serum and oxygen levels are presented, and the potential for manipulating the environment of this system for studying mechanisms of cutaneous development is also discussed.
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  • 94
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    Cell & tissue research 192 (1978), S. 359-361 
    ISSN: 1432-0878
    Keywords: Spermatids ; Human ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Osmiophilic granules with surrounding vesicles resembling flower-like structures occur transiently during the differentiation of human spermatids. These organelles are incorporated into the residual bodies when mature spermatids are released from the germinal epithelium.
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  • 95
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    Cell & tissue research 281 (1995), S. 69-76 
    ISSN: 1432-0878
    Keywords: Periodic lamellar granule ; Primary granule ; Chronic myeloproliferative disorder ; Neutrophils ; Electron microscopy ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Granules consisting of periodically arranged membranous lamellae and amorphous electron-opaque material, i.e., periodic lamellar granules, are present in human neutrophils. To date, no extensive ultrastructural studies have been carried out on these granules because of their infrequent presence in neutrophils. The bone marrow of 18 cases of chronic myeloproliferative disorders, including one case of chronic neutrophilic leukemia in which periodic lamellar granules were frequently seen in neutrophils, was investigated by electron microscopy. Periodic lamellar granules were seen in neutrophils in 12 of the 18 cases at varying frequencies. They were preferentially seen in immature neutrophils. The transverse profiles of these granules revealed concentric complete/incomplete rings or periodic parallel straight lines, i.e., various patterns of lamellar arrangement were present. Periodic lamellar granules were positive for myeloperoxidase and lysozyme at the electron-microscopic level. These results suggest that these granules represent a primary neutrophil granule subtype. However, their functional and pathologic significance remains unknown.
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  • 96
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    Cell & tissue research 280 (1995), S. 183-188 
    ISSN: 1432-0878
    Keywords: Key words: Blood platelets ; Oxidized LDL ; Cytochalasin D ; Reflection contrast microscopy ; Image analysis ; Electron microscopy ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The adhesion of human blood platelets is studied with an in vitro model using reflection contrast microscopy and an image analysis system. The adhesive feature is promoted by oxidatively modified low density lipoprotein, which also induces functional morphological changes of platelets. However, when washed platelets are pretreated with 0.05 mM cytochalasin D, oxidized low density lipoprotein (100 μg/ml) causes a slower increase of the adhesion area (11.6 μm2/min) compared to untreated platelets (15.7 μm2/min) or platelets treated by oxidized low density lipoprotein alone (20.5 μm2/min, P〈0.01). These results are supported by light transmission analysis and by transmission electron microscopy. Our experiments suggest that cytochalasin D inhibits the change of platelets in shape induced by oxidized low density lipoprotein, hinders the adhesion, but does not prevent the adhesion entirely.
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  • 97
    ISSN: 1432-0878
    Keywords: Key words: Respiratory epithelium ; Cuboidal metaplasia ; Cultured cells ; Explants ; Hexamethylene bisacetamide ; Dimethylsulfoxide ; Retinoic acid ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The alterations in the mucociliary unit in the course of chronic inflammation of the upper respiratory tract correspond to morphologic anomalies of the respiratory epithelium and induce cuboidal and squamous metaplasia. While the squamous pattern is most probably irreversible, it is still not clear whether it is possible to restore ciliogenesis in cuboidal metaplasia. In the present study, the action of different inductors of differentiation was evaluated in vitro in isolated cells and explants from human nasal metaplastic epithelium. Polar/apolar compounds induced secretory activity, whereas retinoic acid was able to induce ciliogenesis in some cases. Therefore, the cuboidal metaplastic condition appears to be reversible, and two distinct pathways of differentiation, secretory and ciliogenetic, are identifiable.
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  • 98
    ISSN: 1432-0878
    Keywords: Microgravity ; Collagen ; Bone ; Dermal fibroblasts ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Astronauts experiencing long periods of space flight suffer from severe loss of bone tissue, particularly in those bones that carry the body weight under normal gravity. It is assumed that the lack of mechanical load decreases connective tissue biosynthesis in bone-forming cells. To test this assumption, quantitative and qualitative aspects of collagen synthesis under microgravity, normal gravity, and hypergravity conditions were investigated by incubating human fibroblast cultures with [3H]-proline for 4, 7, 10, and 20 h during the Spacelab D2-mission in 1993. Quantitative analysis revealed an increase of collagen synthesis under microgravity conditions, being up to 143% higher than in 1 g controls. In contrast, hypergravity samples showed a decrease in collagen synthesis with increasing g, being at the 13% level at 10 g. The relative proportion of collagen in total synthesized protein showed a slight decrease with increasing g. The secretion of collagen by the cells, proline hydroxylation of individual collagen α-chains, and the relative proportions of synthesized collagens I, III, and V were not affected under any of the applied conditions.
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  • 99
    ISSN: 1432-0878
    Keywords: Key words: Elastic fibres ; Placental stem villi ; Extravascular smooth muscle cells ; Adhesion plaques ; Talin immunoreactivity ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The stroma of human placental stem villi is believed to consist only of reticular and collagen fibres. In the present study we were able to show for the first time by light (orcein staining) and electron microscopy large amounts of elastic fibres in the stem villous stroma. Electron microscopically, homogeneous elastin was found alone or in association with microfibrils. In addition, microfibrils were observed forming long bands. These three structures, generally known to form elastic connective tissue, were seen in close connection with placental extravascular smooth muscle cells, which belong to the perivascular contractile sheath (PVCS) of stem villi. Elastin was associated with these smooth muscle cells and connected to collagen fibres via microfibrils. Collagen fibres were additionally interconnected by spike-like structures. Extravascular smooth muscle cells revealed numerous adhesion plaques which occupied conspicuously long cytoplasmic faces of the plasma membrane. In cryostat sections, immunoreactivity of talin, an attachment protein of adhesion plaques linking intracellular α-actin filaments with extracellular fibronectin, was detected in extravascular and vascular (media) smooth muscle cells. The arrangement of placental extravascular smooth muscle cells, elastic and collagen fibres suggests a functional myofibroelastic unit within the PVCS, which surrounds the large foetal blood vessels possibly contributing to elasticity and supporting tensile and/or contracting forces within the stem villi.
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  • 100
    ISSN: 1432-0878
    Keywords: Key words: Cell adhesion molecules ; neuronal ; Stellate cells ; Liver ; Immunohistochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Neural cell adhesion molecule (N-CAM) is distributed in most nerve cells and some non-neural tissues. The present immunohistochemical study has revealed, for the first time, the expression of N-CAM in perisinusoidal stellate cells of the human liver. Liver specimens were stained with monoclonal antibody against human Leu19 (N-CAM) by a streptoavidin-biotin-peroxidase-complex method. Light- and electron-microscopic analyses have shown that N-CAM-positive nerve fibers are distributed in the periportal and intermediate zones of the liver lobule. Perisinusoidal stellate cells in these zones are also positive for N-CAM. N-CAM is expressed on the surface of the cell, including cytoplasmic projections. Close contact of N-CAM-positive nerve endings with N-CAM-positive stellate cells has been observed. On the other hand, stellate cells in the centrilobular zone exhibit weak or no reaction for N-CAM. Perivascular smooth muscle cells and fibroblasts in the portal area and myofibroblasts around the central veins are negative for N-CAM. The present results indicate that the perisinusoidal stellate cells in the periportal and intermediate zones of the liver lobule characteristically express N-CAM, unlike other related mesenchymal cells, and suggest that the intralobular heterogeneity of N-CAM expression by stellate cells is related to the different maturational stages of these cells.
    Type of Medium: Electronic Resource
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