The intima: historic literature revisited.
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Anatomy↗
Biomedical subjects
A Draeger
Publications and source records attributed to A Draeger.
Recommendations for muscle research in space.
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Humans↗
[Comparison of the results from commercially available Brucella ELISA test kits for the investigation of bovine sera].
Bovine sera from a farm with bacteriologically confirmed brucellosis were taken for comparative serological studies using conventional methods (slow agglutination test--SAT, complement fixation test--CFT, Rose-Bengal-plate-agglutination-test--RBPT) and 5 different commercially available brucellosis ELISA testkits. These serum samples together with other infectious sera producing low level positive antibody titers in conventional tests did not react uniformly when tested by five different commercial ELISA testkits. The introduction of a standard control serum with a general cut-off resulted in a greater uniformity of ELISA results from 4 of 5 ELISA testkits and a closer approach to results of the CFT which is regarded as the confirmatory test for brucellosis. This control serum, on the other hand, was not suitable for the identification of the sensitivity of the various ELISA test kits. For this purpose, in SAT and CFT weakly positive reacting brucella sera from infected farms is needed.
Agglutination Tests↗
Human foetal lung (IMR-90) cells: myofibroblasts with smooth muscle-like contractile properties.
Single cells displaying many characteristics in common with smooth muscle are now being identified in many organs. Although their origin remains elusive, they are nonetheless known to play a major role in fibroproliferative diseases and stromal reactions by virtue of their contractile properties. We have investigated the contractile properties and expression of smooth muscle-associated proteins in such a cell line derived from human foetal lung (IMR-90). For almost two decades, these cells have served as model fibroblasts in a wide variety of studies. And yet, IMR-90 cells manifest many features characteristic of differentiated smooth muscle cells: they exhibit the same elongated, slender form and the strikingly regular, longitudinal alignment of their actin-attachment sites, which are indispensable for coordinated contraction. Moreover, these adhaerens junctions also contain metavinculin, the smooth muscle analogue of vinculin, the stable expression of which has not previously been demonstrated in cultured cells. Since sm 22, as well as the smooth muscle-associated alpha-actin isoform and calponin are also expressed, IMR-90 cells must now be classified as myofibroblasts. When cultivated on a rubbery silicon surface, IMR-90 cells respond to stimulation with a rate of contraction which is considerably faster than that exhibited by fibroblasts derived from other established lines. Taken together, the regular longitudinal orientation of the adhaerens junctions, the stable expression of metavinculin, and the rapid speed of shortening in IMR-90 cells suggest, by implication, that the periodicity of actin attachment sites is a fundamental determinant of contractile efficiency in smooth muscle cells; this spacing may be mediated by metavinculin.
3T3 Cells↗
Vascular smooth muscle cells of H-2Kb-tsA58 transgenic mice. Characterization of cell lines with distinct properties.
BACKGROUND: The vascular wall is composed of at least two different populations of smooth muscle cells that are distinct in their structure and protein composition. According to the developmental stage of tissue taken for culture, the ratio between cells of epithelioid phenotype and spindle-shaped cells is variable. In particular, the epithelioid cells display characteristic features associated with immaturity. Because their increased appearance can be observed in endothelial denudation, the represent a dedifferentiated, proliferative smooth muscle cell type with a repair function in vascular injury. METHODS AND RESULTS: To investigate this cellular heterogeneity, we established vascular smooth muscle cell lines from H-2Kb-tsA58 transgenic mice. Due to temperature-sensitive expression of the SV 40 large T-antigen in cells derived from this mouse strain, our smooth muscle lines were conditionally immortalized from the onset of their life in culture. Thus, we were able to clone cell lines representing the two different phenotypes described so far. Epithelioid cells derived from newborn animals are characterized by their expression of cytokeratins and the development of tight junctional complexes. Spindle-shaped cells, which could be isolated from newborn or adult animals, corresponded in phenotype and protein expression to smooth muscle cell lines established previously. CONCLUSIONS: The special properties of vascular smooth muscle cells of the epithelioid phenotype suggest an endothelial replacement function in the course of injury to the vascular wall.
Actins↗
Heterogeneity of smooth muscle-associated proteins in mammalian brain microvasculature.
In the brain, the microvascular system is composed of endothelial cells surrounded by a layer of pericytes. The lack of smooth muscle cells in this tissue suggests that any contractile function must be performed by one or both of these cell types. The present study was undertaken in order to identify cells in terminal blood vessels that contain smooth muscle-like contractile machinery. Endothelial cells were reactive with antibodies against smooth muscle myosin but showed no other smooth muscle-related features. In contrast, pericytes of intact microvessels showed a pattern of protein expression similar to that of smooth muscle cells. Pericytes also behaved in tissue culture like cultured smooth muscle cells, with regard to the changes in expression of smooth muscle-related proteins. These data confirm the close relationship between smooth muscle cells and pericytes, and point to their contractile function in the brain microvessels.
Animals↗
Smooth muscle differentiation in human myometrium and uterine leiomyoma.
Smooth muscle differentiation has been analysed in human myometrium and leiomyoma by Western blotting with antibodies to smooth muscle specific proteins. No differences in the expression of h-caldesmon, metavinculin, desmin, alpha-smooth muscle actin and calponin were observed. The technique of two-dimensional gel electrophoresis was used, therefore, to further analyse differences between normal smooth muscle cells and their neoplastic counterparts. By comparing the protein patterns of normal myometrium and leiomyoma, it was possible to identify a protein with a molecular weight of approximately 27 kD that is selectively expressed in normal uterine smooth muscle cells. This protein proved to be a low molecular weight variant of calponin, a smooth muscle specific protein of as yet unknown function. Its immediate downregulation in tissue culture of normal myometrium points to a possible role in the process of dedifferentiation.
Adult↗
Calponin. Developmental isoforms and a low molecular weight variant.
Two-dimensional gel analysis of basic proteins in developing human smooth muscle identifies calponin as a prominent marker of the differentiated phenotype. Adult tissue (human and mouse) typically expresses up to four calponin isoforms, three of which appear sequentially during fetal development: adult myometrial cells express the same three isoforms in primary culture in vitro and these are down-regulated, in reverse order, during the subsequent modulation of phenotype. Monospecific, polyclonal antibodies against calponin identify a lower molecular weight variant of calponin (L-calponin) that is strongly and specifically expressed in adult smooth muscles of the human urogenital tract. L-calponin is down-regulated in benign smooth muscle derived tumors (leiomyoma) and is not expressed in primary cultures of normal uterine tissue.
Animals↗
Human acardiac anomaly: a report of three cases.
We report on three cases of acardia, all of which had to be classified as acardius anceps. The degree of cardiac malformation ranged between a completely missing heart and a malrotation of a relatively well-developed heart. One fetus showed an intermediate form of cardiac malformation with a hypoplastic cor triloculare. Our findings suggest that the defect in cardiac development may take place at different times in early embryogenesis, resulting in differing degrees of cardiac malformation.
Abnormalities, Multiple↗
Cytokeratins, smooth muscle actin and vimentin in human normal salivary gland and pleomorphic adenomas. Immunohistochemical studies with particular reference to myoepithelial and basal cells.
The distribution of immunostaining in normal major salivary gland and in 12 pleomorphic adenomas was studied using monospecific monoclonal antibodies to a number of cytokeratins, including cytokeratin 14, to smooth muscle actin and vimentin. A number of these antibodies enabled a distinction to be made between structural components of the normal gland, and to relate this to the different structures of pleomorphic adenomas. In the normal gland, the luminal duct cells expressed cytokeratins 7, 8, 18 and 19. Three antibodies were of particular value for the characterization of normal myoepithelial and basal cells; while the antibody to smooth muscle actin and the cytokeratin antibody Ks8.12 mutually exclusively stained the myoepithelial (basket) cells and the basal duct (light) cells, respectively, the recently established monospecific antibodies to cytokeratin 14 showed specific immunostaining with both cell types. These three antibodies left luminal cells virtually unstained. Ck 13 was found occasionally in single luminal excretory duct cells. Antibodies to cytokeratins 1/2, 10 and 10/11 did not show any staining in the normal gland. In the pleomorphic adenomas, the staining pattern of the two-layered tubular formation resembled that of the normal gland ducts: tumour luminal cells showed the characteristic, although more irregular, expression of cytokeratins 7, 8, 18 and 19; the outer cells resembled normal ductal basal cells with their anti-cytokeratin 14/Ks8.12-epitope staining and in that they virtually lacked staining for smooth muscle actin. Trabecular formations and cells in myxoid areas were reactive with Ks8.12 and for cytokeratin 14, occasionally also for cytokeratins 7, 18 and 19. Epidermoid cell islets expressed mainly cytokeratin 14 and inconsistently the squamous epithelial cytokeratin 13 and the epidermal cytokeratin 10/11. Vimentin was found in cells of myxoid areas. The results support the postulate that some of the normal duct basal cells act as reserve cells and can give rise to tumour formation with a primitive myxoid or trabecular pattern and a more differentiated tubular or epidermoid configuration.
Actins↗
Supercontracted state of vertebrate smooth muscle cell fragments reveals myofilament lengths.
Isolated cell preparations from chicken gizzard smooth muscle typically contain a mixture of cell fragments and whole cells. Both species are spontaneously permeable and may be preloaded with externally applied phalloidin and antibodies and then induced to contract with Mg ATP. Labeling with antibodies revealed that the cell fragments specifically lacked certain cytoskeletal proteins (vinculin, filamin) and were depleted to various degrees in others (desmin, alpha-actinin). The cell fragments showed a unique mode of supercontraction that involved the protrusion of actin filaments through the cell surface during the terminal phase of shortening. In the presence of dextran, to minimize protein loss, the supercontracted products were star-like in form, comprising long actin bundles radiating in all directions from a central core containing myosin, desmin, and alpha-actinin. It is concluded that supercontraction is facilitated by an effective uncoupling of the contractile apparatus from the cytoskeleton, due to partial degradation of the latter, which allows unhindered sliding of actin over myosin. Homogenization of the cell fragments before or after supercontraction produced linear bipolar dimer structures composed of two oppositely polarized bundles of actin flanking a central bundle of myosin filaments. Actin filaments were shown to extend the whole length of the bundles and their length averaged integral to 4.5 microns. Myosin filaments in the supercontracted dimers averaged 1.6 microns in length. The results, showing for the first time the high actin to myosin filament length ratio in smooth muscle are readily consistent with the slow speed of shortening of this tissue. Other implications of the results are also discussed.
Actin Cytoskeleton↗
The cytoskeletal and contractile apparatus of smooth muscle: contraction bands and segmentation of the contractile elements.
Confocal laser scanning microscopy of isolated and antibody-labeled avian gizzard smooth muscle cells has revealed the global organization of the contractile and cytoskeletal elements. The cytoskeleton, marked by antibodies to desmin and filamin is composed of a mainly longitudinal, meandering and branched system of fibrils that contrasts with the plait-like, interdigitating arrangement of linear fibrils of the contractile apparatus, labeled with antibodies to myosin and tropomyosin. Although desmin and filamin were colocalized in the body of the cell, filamin antibodies labeled additionally the vinculin-containing surface plaques. In confocal optical sections the contractile fibrils showed a continuous label for myosin for at least 5 microns along their length: there was no obvious or regular interruption of label as might be expected for registered myosin filaments. The cytoplasmic dense bodies, labeled with antibodies to alpha-actinin exhibited a regular, diagonal arrangement in both extended cells and in cells shortened in solution to one-fifth of their extended length: after the same shortening, the fibrils of the cytoskeleton that showed colocalization with the dense bodies in extended cells became crumpled and disordered. It is concluded that the dense bodies serve as coupling elements between the cytoskeletal and contractile systems. After extraction with Triton X-100, isolated cells bound so firmly to a glass substrate that they were unable to shorten as a whole when exposed to exogenous Mg ATP. Instead, they contracted internally, producing integral of 10 regularly spaced contraction nodes along their length. On the basis of differences of actin distribution two types of nodes could be distinguished: actin-positive nodes, in which actin straddled the node, and actin-negative nodes, characterized by an actin-free center flanked by actin fringes of 4.5 microns minimum length on either side. Myosin was concentrated in the center of the node in both cases. The differences in node morphology could be correlated with different degrees of coupling of the contractile with the cytoskeletal elements, effected by a preparation-dependent variability of proteolysis of the cells. The nodes were shown to be closely related to the supercontracted cell fragments shown in the accompanying paper (Small et al., 1990) and furnished further evidence for long actin filaments in smooth muscle. Further, the segmentation of the contractile elements pointed to a hierarchial organization of the myofilaments governed by as yet undetected elements.
Actinin↗
Unique geometry of actin-membrane anchorage sites in avian gizzard smooth muscle cells.
Adherens junctions in isolated avian gizzard smooth muscle cells appear as short longitudinal streaks or chevrons that are arranged in periodic, mainly transverse bands along the cell surface. This barrel-like geometry, revealed by antibodies to either vinculin or talin, was seen also in teased gizzard strips by confocal laser-scanning microscopy and contrasted with the rib-like surface pattern observed here and previously in other avian and mammalian smooth muscles. There were on average 67 transverse bands per gizzard cell and an estimated total of around 800 vinculin/talin sites. The longitudinal spacing between the transverse bands of vinculin streaks in the gizzard cells changed from 4-5 microns in extended cells to around 1 micron in shortened cells and the bands remained essentially transverse at all cell lengths, inconsistent with a screw-like mode of cell shortening as has been invoked for smooth muscle cells by others. The absence of rotation on shortening was confirmed by observations on isolated and bead-decorated skinned cells that were induced to contract with ATP. Counterlabelling of cells with alpha-actinin antibodies produced more or less exclusive staining of the cytoplasmic dense bodies, and little surface label: the total number of dense bodies per cell, estimated from confocal microscope through focal series was in the range of 3000. The data are consistent with a periodic anchorage of actin filaments to the cell surface and, in turn, with the existence of regularly spaced contractile assemblies.
Animals↗
The amniotic adhesion malformation complex associated with short or absent umbilical cord.
The spectrum of amniotic adhesion malformations can include body wall defects, cranial abnormalities and amputation of limbs. Its greatly varying pattern renders diagnosis difficult. Pathogenesis has been ascribed to rupture of amnion in early gestation with subsequent entanglement of fetal parts (Torpin R, Am J Obstet Gynecol 1965;91:65-75 and in Fetal malformation caused by amnion rupture during gestation, Springfield, IL: Charles C. Thomas, 1968). Eight cases belonging to the amniotic adhesion malformation complex which occurred within a 7 months period were seen in our department. This malformation complex is not infrequently observed; however, this clustering of cases is exceptional. A common feature in all subjects was an either missing or significantly shortened umbilical cord, whereas amniotic bands could only be identified in two fetuses.
Abnormalities, Multiple↗
[Amniotic band syndrome associated with an acardiac malformation in a twin pregnancy. Apropos of a case].
We report herein a case of a twin pregnancy with one fetus showing an amniotic band syndrome, the other being an acardiac monster. This is, to our knowledge, the first case of this unusual combination. Both fetuses showed multiple severe malformations. According to the classification of Napolitani, one fetus had to be classified as "acardius anceps". The malformations of both bodies and the anomalies of both umbilical cords suggest a similar defect of the germinal disc, possibly occurring very early during the pregnancy.
Abnormalities, Multiple↗
Primary, secondary and tertiary myotubes in developing skeletal muscle: a new approach to the analysis of human myogenesis.
Monoclonal antibodies to myosins have been used to describe and define the appearance and maturation of 3 different classes of myotube in developing human quadriceps muscle. Five monoclonal antibodies were used: (i) MAb A against human slow myosin heavy chain; (ii) MAb B against a myosin heavy chain present in most adult Type 2 fibres; (iii) MAb C against myosin heavy chain present in all mature and immature Type 2 fibres; (iv) MAb D, with similar reactivity to MAb C; (v) MAb E against human embryonic myosin. The combined use of two of these antibodies (A and B) enables the confident early identification of each of 3 classes (primary, secondary, tertiary) of myotubes, which appear sequentially during myogenesis. Our results show that induction of slow myosin heavy chain synthesis is a biphasic phenomenon in developing human skeletal muscle. Slow myosin heavy chain was present in all the earliest (9 weeks gestation) primary myotubes, but was not detected in secondary or tertiary myotubes until about 29 weeks gestation. Each stage of fetal muscle development has a characteristic immunocytochemical pattern which reveals cellular heterogeneity not evident on myosin ATPase histochemistry. Myosin immunocytochemistry may usefully be applied to assess the gestational age of fetuses. A new interpretation of human skeletal muscle development is proposed, based on the separate programming of 3 different kinds of myotube. This may be important in the analysis of diseased muscle in which developmental abnormalities or regeneration are present.
Antibodies, Monoclonal↗
[Otitis pubis in an adolescent].
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Adolescent↗
[Calcium-dependent association of membrane rafts].
The plasma membrane of eucariotic cells is composed from spatially distinct regions enriched in sphingomyelin and cholesterol (so called membrane rafts), which are surrounded by glycerophospholipid milieu. The ability of these regions (membrane microdomains) to recruit specific enzymes and structural proteins results in spatial segregation of elements of different signaling systems. To the great extent, such segregation is a result of membrane raft association. Our results demonstrate that association of individual rafts is a controlled process, which is regulated, as many other biological processes, by Ca2+. Annexin II, a member of phospholipid-binding protein family, interacts with preparations of membrane rafts in a Ca(2+)-dependent manner thus resulting in reversible raft-association. Occurring in the presence of physiologically relevant calcium concentrations these interactions may play a role in regulation of smooth muscle contraction.
Animals↗