Adaptive remodelling of smooth muscle in the neo-intima of vein-to-artery grafts in rats: a detailed morphometric analysis

Summary End-to-end autogenous vein-to-artery grafts in rats have been used extensively as a model for neointimal thickening (hyperplasia), which develops over the first 6 weeks after grafting. This study employed computerised morphometric techniques to analyse 16 grafts, in order to quantitate precisely how the neointima develops. Two important features were described that have not been identified previously, due to the extensive variation in neo-intimal thickness inherent in vein grafts. Firstly, the proximal region of the graft was significantly thicker than the distal region, up until 6 months after grafting. The smooth muscle cells in the graft may have developed more rapidly in the proximal region, due to the altered haemodynamics within the graft. Secondly, within the central region of the graft the characteristic focal nature of neo-intimal hyperplasia was evident throughout the period of the study, but by 6 months the neo-intima tended to be distrubuted more evenly. By 6 months remodelling of smooth muscle throughout the graft neo-intima had occurred, and the neo-intima had matured to a thickness equivalent to that of the intima plus media of the adjacent iliac artery.     

Influence of conventionalization on cecal wall structure of germ-free Wistar rats: quantitative light and qualitative electron microscopic observations

Summary The structural changes of the cecal wall in germfree rats were observed at regular intervals after the inoculation of fecal microflora from conventional rats. Quantitative light microscopy showed that most of the elements in the cecal wall increased at 12 or 24 h and reached peak values at 4 days after inoculation. On the 7th day, they decreased approximately to the values for conventional rats. The crypts were bent or widely open till 24 h but were not after the 4th day. Hyperplasia of the crypt epithelial cells including mucous-type cells was observed following microbial inoculation. Electron microscopy revealed that most of the epithelial cells lining the mucosa were typical columnar cells. Desquamation of the epithelial cells and contraction of the muscle fibers were often seen on 4th day. The mucous-type cells were divided into two types, goblet and non-goblet mucous-type cells. Reduction of cecal volume after microbial inoculation may be mainly caused by muscle contraction in the early period and hyperplasia and desquamation of the epithelial cells may suggest their role as the first and non-specific defense line prior to operation of the specific immune system.     

Quantitative electron microscopic observations on Paneth cells of germfree and ex-germfree Wistar rats

Summary Ultrastructural changes of Paneth cells of germfree (Gf) rats which had been inoculated with bacteria-containing feces from conventionally-reared (SPF) rats were quantitatively examined. 12 and 24 h after inoculation, the Paneth cells showed a striking decrease in the number of secretory granules and the occurrence of large vacuoles. Phagosomes containing bacteria were not seen. After 4 days, the secretory granules reaccumulated and smooth-surfaced apical vesicles increased in number. It is discussed that the large vacuoles may be related to membrane-retrieval events following the massive extrusion of secretory granules whereas the apical vesicles appear to serve this function when exocytosis is not pronounced. In addition to the large secretory granules ca. 10% of Paneth cell profiles contained a few dense-cored vesicles measuring about 150 nm in diameter which resemble peptidergic neurosecretory granules.     

Lung of the tree frog, Hyla arborea L. A scanning and transmission electron microscopic study

Summary Lungs ofHyla arborea L. were examined by scanning and transmission electron microscopy and morphometric methods. The lungs contain several interconnected folds in a netlike reticular arrangement of first, second and third order, mainly covered with pneumocytes. On the septa of first and second order, irregularly distributed small patches of ciliated epithelium devoid of goblet cells are located. Dome-shaped neuroepithelial bodies can be seen in the vicinity of ciliated epithelium. The pulmonary epithelium consists of one type of pneumocyte, which contain in their cytoplasm three kinds of bodies: lamellar, dense and multivesicular. The dense bodies are precursors of lamellar bodies (LBs), while the multivesicular bodies are incorporated into the LBs, being later secreted to the air space. The lining layer covering the internal lung surface contains numerous transformed LBs but tubular myelin figures are scarce. The surface of the lining layer is coated by a thin film. The air-blood barrier, consisting of three layers: epithelium, interstitial space and endothelium, is 0.6 m thick.     

Light and electron microscopic examination of endothelial cells from bovine retinal vessels in long-term cultures

The aim of the present work was to examine and compare the ultrastructure of bovine retinal endothelial cells (BRECs) in vitro during several passages in a medium selective for endothelial cells. The identity of the endothelial cells was confirmed immunohistochemically, up to the tenth passage. Changes in their ultrastructure in comparison to endothelial cells in vivo occurred at the onset of culturing and not progressively with repeated passages. The cultured BRECs show high metabolic activity in all passages. While retaining their identity as endothelial cells, they modify their lipid metabolism, so that lipids are stored. This change in lipid metabolism was induced by the medium. Received: 19 May 1999 / Accepted: 4 August 1999     

Spleen alterations in hairy cell leukemia: a scanning electron microscopic study

In Hairy Cell Leukemia (HCL) peripheral blood and bone marrow cells show under the scanning electron microscope (SEM) a characteristic surface with numerous ruffles and microvilli. The spleen of a patient affected by HCL was studied by SEM after fresh sectioning and routine preparation. Cells with the typical "hairy" surface were observed infiltrating the red pulp, altering the normal reticular meshwork and causing red blood cell distortion. In the sinuses, hairy cells adhered to the endothelial cells causing sinus dilatation and destruction. Aggregates of hairy cells delimiting pooled erythrocytes were also observed and may represent the "pseudosinuses" described in previous light and transmission electron microscopic studies. These preliminary findings may explain the condition of hypersplenism which characterizes HCL. In addition, SEM is proposed as a rapid and simple method to identify HCL spleen involvement.     

Age-related distribution of endocrine cells in the human prostate: a quantitative study

A morphometric analysis was performed to obtain quantitative data on age-related changes in prostatic endocrine cell (PrEC) density. Sixty prostates from subjects aged 14–74 years were studied with a semi-automatic image analysis system (ASM 68K, Leitz) applied to sections immunostained for chromogranin A-reactive cells. The highest density of PrECs (0.366 cells/mm of epithelial length) was found in the 25–54 year age group, which was significantly different from that found in prostates of the younger (0.311 cells/mm) and the older (0.261 cells/mm) age groups. The data probably reflect the higher incidence of incompletely developed glandular units in the younger group and the formation of new alveoli related to the usual glandular hyperplasia that occurs with increasing age in the older group.     

The ciliary ganglion of the cat: a light and electron microscopic study

The ciliary ganglia of eight healthy adult cats were studied by light and electron microscopy. The ganglion, measuring about 2 mm in length, was consistently found to be attached to the branch from the oculomotor nerve supplying the inferior oblique muscle. The number of neurons varied from 2773 to 3794 after applying Abercrombie's correction. The mean of average somal diameter of the neurons was 36.5 m (SD = 5.0 m) and the mean of somal cross-sectional area was 904.2 m² (SD = 262.8 m²). The mean of average nuclear diameter was 13.9 urn (SD = 1.8 m) and the mean of nuclear cross-sectional area was 142.2 m² (SD = 37.1 m²). The mean of the aspect ratios of the soma and nucleus were 1.2 (SD = 0.1) and 1.1 (SD = 0.1) respectively. The frequency distributions of these parameters were all unimodal. Under the light microscope, the Nissl granules in the neurons were prominent and were distributed peripherally, perinuclearly or randomly in the cytoplasm. Under the electron microscope, the rough endoplasmic reticulum showed a similar pattern of distribution in the cytoplasm. In some neurons, glycogen-like granules were present; these were either distributed randomly throughout the cell, or aligned in single rows in relation to sub-surface cisterns and between the cisterns of smooth and rough endoplasmic reticulum. Most of the dendrites were short protrusions from the cell body; some contained glycogen-like granules. Occasionally, the dendritic protrusions were electron-dense. All the synapses encountered were axodendritic. In most axon terminals, the synaptic vesicles were spherical and measured 30–50 nm in diameter; in some, they were flattened, measuring 50 nm by 20 nm. Some axon terminals containing either spherical or flattened synaptic vesicles also contained large dense-cored vesicles that measured 80–100 nm, while their dense core measured 40–60 nm.     

Muscle spindles in immobilized muscle: electron microscopic study of recovery

This study is concerned with the morphological recovery process of muscle spindles following a long period of immobilization. The right hindlimbs of rats were fixed with a plaster cast for 4 weeks. Thereafter, four groups of rats were examined by electron microscopy. One group served as the control after the cast was removed. The other three groups were examined after free walking for 4, 8, and 12 weeks, respectively. The muscle spindles (tibialis anterior muscle) of the individual animals were then ultrastructurally analyzed. The morphological alterations (of the outer capsule, intrafusal muscle fibers, and intrafusal nerve components) gradually recovered during free walking and regained almost all normal features in 12 weeks after returning to walking.A part of this research was presented at the 25th Annual Meeting of the Clinical Electron Microscopy Society of Japan, Matsumoto, September 28–30, 1993.     

Lipofuscin in the cerebellar cortex of albino rats: An electron microscopic study

Summary The ultrastructure of autofluorescent, PAS-positive lipofuscin in Purkinje, granule, Golgi epithelial, basket and stellate, microglial and perivascular cells in the cerebellar cortex of senescent rats is described. The membrane-bounded pigment is composed of three elements: 1) electron-lucent homogeneous droplets, 2) a granular matrix and 3) intensely osmiophilic patches. The proportions of these three components vary between cell types and one can grossly differentiate a neuronal and a glial lipofuscin. The lipofuscin granules of stellate and perivscular cells are different from lipofuscin of other cerebellar neurons and glia. It can be concluded from these morphological observations that each cerebellar cell type has its distinct lipofuscin.Supported by the Deutsche Forschungsgemeinschaft La 184/5I would like to thank Mrs. v. Bronewski and Mr. H. Boffin for their technical assistance     

母体急性淋巴细胞白血病胎盘转移的电镜观察

目的　探讨胎盘防止母体 胎儿恶性肿瘤转移的屏障作用。方法　利用透射电镜研究 2例急性淋巴细胞白血病患者胎盘转移的超微结构。结果　在胎盘绒毛表面纤维蛋白沉积物中可见肿瘤细胞、溶解的细胞及细胞碎片。纤维蛋白沉积处及其附近常见合胞体脱落 ,并可见肿瘤细胞粘附于绒毛滋养层基底膜上。合胞体内有些肿瘤细胞基本保持完整的形态 ,有些则出现变性或坏死现象。结论　胎盘能杀伤和分解转移的肿瘤细胞 ,进而防止母体 -胎儿恶性肿瘤转移。     

Immunohistochemical and electron microscopic studies of Langerhans cells in a case of multiple eccrine spiradenomas

Multiple eccrine spiradenomas are rare. In the present study, a detailed investigation of eccrine spiradenoma was performed, focusing in particular on the presence of Langerhans cells (LCs) in the tumor, and their immunohistochemical and ultrastructural characterization. The patient was a woman in her mid-forties who underwent resection of two tumors of the head that were 2.0 and 0.7 cm in size. They were diagnosed as eccrine spiradenoma and were composed of small and large tumor cells with a dense fibrous capsule in the dermis. Immunohistochemically, staining by antibodies to cytokeratins (AE1/AE3, CAM5.2) and CK 5/6 was diffusely positive in all tumor cells, although not in intermingled LCs, which harbored interdigitated nuclei. The cytoplasm of LCs was positive for S-100 protein and CD1a, and their nuclei were also occasionally positive for S-100 protein. Antibody to epithelial membrane antigen was positive for the surface of both intracytoplasmic and true glandular lumina. Fine structural examination revealed the presence of LCs among the tumor cells, extending fine irregular processes among the tumor cells. Birbeck granules were clearly demonstrated in the cytoplasm of LCs. Other fine structural findings included intracytoplasmic lumina with microvilli on their surfaces in some tumor cells. In these examinations of eccrine spiradenoma, LCs, approximately 15/HPF in the tumor, were distinctly detected even at light microscopic level as negative for various types of cytokeratin stains, although they were positive for S-100 protein and CD1a, whereas on ultrastructural examination Birbeck granules were demonstrated in their cytoplasm. Determination of the significance of these LCs in eccrine spiradenoma requires further investigation of a larger number of cases.     

Pathophysiological effects of fibrin on arterial endothelial cells in vivo: an electron microscopic study

Interactions between fibrin and arterial endothelial cells of rat iliac arteries in vivo were studies electron microscopically using a newly devised method. The microvilli became attached to the fibrin threads in an initial period of fibrinolysis. These threads had a fine granular appearance in the lysed areas. Fibrinolytically active endothelial cells had an active vesicular transport for ferritin particles injected concomitantly with the fibrinogen-thrombin mixture, thereby implying the enhancement of endothelial permeability in the lysed areas. However, fibrinolytically inactive endothelial cells coexisted in the same arteries. The denuded intima showed no lytic changes in the fibrin threads. These results indicate that the microvilli of the endothelial cells may plan an important role in the releasing plasminogen tissue activator from the endothelial cells and that there is a heterogeneity with regard to reactivity of each endothelial cell to fibrin.     

Angiogenic histogenesis of stromal cells in hemangioblastoma: ultrastructural and immunohistochemical study

Controversy regarding the origin of characteristic stromal cells (SC) is responsible for the placement of hemangioblastoma as a single entity in the category of "tumors of uncertain histogenesis" in the current WHO classification of brain tumors. This subclassification of hemangioblastoma is, to a large extent, a consequence of a remarkable antigenic heterogeneity of SC demonstrated in many, often contradictory immunohistochemical studies. In contrast, most of the electron microscopic studies demonstrated a number of features indicating angiogenic nature of SC and, therefore, hemangioblastoma. This study reevaluated the histogenesis of SC, applying immunohistochemistry as well as electron microscopy and immunoelectron microscopy. Immunohistochemical studies confirmed most of the previous results indicating a very frequent expression of vimentin, S-100 protein, neuron-specific enolase, and cytokeratins. SC were less commonly immunoreactive for desmin, factor XIIIa, and Ricinus communis lectin receptors, and only occasionally for factor VIII and Ulex europeus lectin. They were negative for other markers of endothelial, neuronal, glial, neuroendocrine, and smooth muscle differentiation. Approximately 1% of SC showed Ki67 immunoreactivity, indicating their slight proliferative activity, consistent with the benign nature of the tumor. In contrast to the inconclusive results of the immunohistochemistry, electron microscopy demonstrated a clear relationship of SC to endothelial cells, smooth muscle cells, and pericytes. Occasional SC were found within the vascular lumina. SC often showed intracellular caveolae consistent with the formation of early capillary lumina. Moreover, occasional SC contained small Weibel-Palade bodies positive for factor VIII in immunoelectron microscopy. SC represent a heterogenous population of abnormally differentiating mesenchymal cells of angiogenic lineage, with some morphological features of endothelium, pericytes, and smooth muscle cells. Occurrence of SC in hemangioblastoma could be related to a limited ability of angioformative stromal cells to develop an architecture of capillary lumina integrated with the vascular network of the tumor. Hemangioblastoma should be reclassified and included together with other vascular tumors of the central nervous system.     

Binding of 125I-endothelin-1 to fat-storing cells in rat liver revealed by electron microscopic radioautography.

Summary Localization of intravenously injected [¹²⁵I]-endothelin-1 was examined in rat liver by light and electron microscopic radioautography. At 10 min after injection, silver grains were localized along the sinusoidal wall, i.e., mostly on the thin processes of fat-storing cells and sinusoidal endothelial cells, and also on the Kupffer cells and the microvilli of hepatocytes. About 35% of the total silver grains were located on the processes of fat-storing cells at 10 min. The grain density (number of silver grains/cell area) of fat-storing cells was three-fold that of Kupffer cells, and 18-fold that of hepatocytes. At 60 min, 60% of the total grains were observed on the fat-storing cells, though the value of grain density was not changed. Silver grains were internalized into the cytoplasm of fat-storing cells and often associated with multivesicular bodies. In contrast, the grain density of endothelial cells and Kupffer cells decreased with time. These results indicate that hepatic fat-storing cells have a considerable number of endothelin-binding sites, and incorporate bound endothelin into cytoplasm.     

Quantitative transmission electron microscopic study of dental plaque--an in vivo study with different mouthrinses

Only a few studies of dental plaque in transmission electron microscopy (TEM) have been performed. The authors have applied TEM for morphometric analysis of dental plaque. Wearing acrylic splints with bovine enamel pieces on the buccal surface of teeth 36 and 46, 21 volunteers rinsed twice a day with a solution of chlorhexidine digluconate, amine/stannous fluoride, or sodium chloride. After 24, 48, and 72 h each a piece of enamel was taken and quantitatively analyzed by TEM for plaque thickness and composition. Similar results were obtained for plaque thickness after rinsing with chlorhexidine digluconate or amine/stannous fluoride. Numerical differences at the counts of cocci or bacilli after rinsing with chlorhexidine digluconate or amine/stannous fluoride did not always have statistical significance.     

Interaction of Listeria monocytogenes with human brain microvascular endothelial cells: an electron microscopic study

Internalization of Listeria monocytogenes into human brain microvascular endothelial cells (HBMEC) has recently been demonstrated to be dependent upon the inlB gene. In the present scanning electron microscopic study we show that L. monocytogenes efficiently interacts with the surface of HBMEC in an inlB-independent manner which is also different from invasion. The inlB-dependent invasion of HBMEC by L. monocytogenes is accompanied by intracellular multiplication, movement, and production of bacterium-containing protrusions. These protrusions extend from the cell surface without perturbation of any adjacent cellular membrane.     

Sexually dimorphic contribution from the fornix to the ventromedial hypothalamic nucleus: a quantitative electron microscopic study

Quantitative electron microscopy confirmed that the neuropil of the ventrolateral part of the ventromedial hypothalamic nucleus (VL-VMHN) of the rat is sexually dimorphic with respect to the density of shaft and axo-spinous synapses, both of which are more numerous in the male. In addition, adult rats with complete interruption of the fornix displayed a sexually dimorphic input in the density of fornical synapses in the neuropil of the VL-VMHN, in which degenerating terminals were more numerous in the male. Perinatal exposure of the female to exogenous testosterone or castration of the newborn male ‘inverted’ these sex differences, demonstrating their hormonal dependence. It is concluded that (1) the fornix provides synaptic input to the VL-VMHN as proven by orthograde degeneration; (2) the number of fornical endings synapsing in the VL-VMHN is greater in the male than in the female; (3) this dimorphism depends of the organizational effect of gonadal sex steroids.     

Changes in proteoglycans of ageing and osteoarthritic human articular cartilage: An electron microscopic study with polyethyleneimine

Background: Ageing and osteoarthritic (OA) cartilage show characteristic alterations in chondrocyte morphology and in the composition and content of matrix proteoglycans (PGs). Data concerning matrix components are mostly of biochemical nature. Ultrastructural histochemistry is needed to gain more information about distribution of these altered matrix components. Methods: We used the cationic dye polyethyleneimine (PEI) to visualize at the EM level alterations in the distribution and dimensions of PGs of human healthy young, healthy aged, and OA articular cartilage. Results: Young cartilage contained PEI-positive granules in the superficial layer and big winding PEI-positive structures in the deeper layers. In the healthy aged tissue, PEI-positive granules were observed throughout the matrix and smaller winding structures were present in the deeper layer. In OA cartilage both types of PEI-positive structures were absent in the superficial layer. Deeper in the matrix PEI-positive granules could be demonstrated. Moreover, PEI-positive angular structures were observed in the deeper zones. Conclusions: The differences in PEI-positive structures are a good reflection of the differences in PGs between young, ageing, and OA cartilage as demonstrated in biochemical studies. PEI, used at the EM level, gave more precise information concerning the localized changes in quality, quantity, and location of PGs in articular cartilage during ageing and disease. © 1994 Wiley-Liss, Inc.     

The ultrastructure of oesophageal carcinomas: multidirectional differentiation. A transmission electron microscopic study of 43 cases.

Forty-three oesophageal carcinomas, comprising 15 squamous cell carcinomas, 22 adenocarcinomas, 5 small cell carcinomas, and 1 adenosquamous carcinoma, were examined by transmission electron microscopy. The ultrastructural features of each tumour type are detailed. Multi-directional differentiation (heterogeneity) was observed in 11 cases: 5 squamous cell carcinomas with focal glandular features; 4 adenocarcinomas with focal squamous features; and 2 small cell carcinomas, one with glandular and the other with squamous features. Abnormal distributions of desmosomes in squamous cell carcinomas and of polarity of tumour cells in adenocarcinomas are described.