Microvascular system of the rat incisor enamel organ. A scanning electron microscopic study of vascular corrosion casts

The rat incisor is a commonly used model in studies of tooth eruption, amelogenesis and effects of mechanical loading on the dental and periodontal tissues. The purpose of this study was to assess the three-dimensional architecture of the microvascular bed of the rat incisor enamel organ, to describe the direction of blood flow, and to provide a histometric assessment of the vascular categories that can be statistically analyzed. Vascular corrosion casts were prepared and examined by scanning electron microscopy. The microvasculature of the labial periodontal space was arranged in three distinct layers. The inner layer in direct relation to the enamel organ consisted of a capillary network which was drained by short venules at the cemento-enamel junction. The intermediate layer consisted of arterioles oriented parallel to the long axis of the incisor mainly mid-labially, branching off smaller arterioles to the capillary network. The outer layer was formed by flattened sinusoid vessels of larger caliber. Blood supply was from the anterior superior alveolar artery branches through the arterioles into the capillary network. Drainage was postero-laterally along the cemento-enamel junction via short venules which emptied into the sinusoid vessels, finally to flow through Volkmann's canals into the alveolar bone via small venules. The findings demonstrate that the microvasculature of the rat incisor enamel organ has an exceptionally high level of physiologically-adapted structural organization.     

The insulo-acinar portal and insulo-venous drainage systems in the pancreas of the mouse, dog, monkey and certain other animals: a scanning electron microscopic study of corrosion casts

Scanning electron microscopy of vascular casts of the pancreas from monkies, cattle, pigs, dogs, cats, rabbits, guinea pigs and mice showed certain species differences in the occurrence of intralobular and interlobular islets and in the microcirculatory pattern of these islets. Interlobularly located islets were frequently found in the mouse and guinea pig, as has been previously established in the rat (Murakami and Fujita, 1992); they emitted insulo-venous efferent vessels directly draining into veins. In contrast, the intralobular islets in the guinea pig usually issued insulo-acinar portal vessels continuous with the lobular capillary network. In the mouse, they usually emitted both the insulo-acinar portal and insulo-venous efferent vessels. The insulo-venous efferent vessels, including those of the interlobular islets, could partly be portal in nature since they occasionally issued portal branches directed to the lobular capillary network. In rabbits, cats, dogs, pigs, cattle and monkies, as in men (Murakami et al., 1992), essentially all islets in the pancreas were intralobular in location and usually emitted the portal vessels only. In the mouse and rabbit, as in the rat (Murakami and Fujita, 1992), the islet received afferent vessels in its superficial aspect and issued efferent vessels from its deep aspect. In the Formosan monkey, as previously reported in the rhesus monkey (Fujita and murakami, 1973), the afferent vessels usually ran deep into the islet which emitted vessels from its superficial aspect. In other animals examined in this study, as in humans (Murakami et al., 1992), no consistent rule concerning the microcirculatory pattern within the islet could be determined.     

Luminal epithelium of Warthin's tumours: a scanning electron microscopic study

Three cases of papillary cystadenoma lymphomatosum were examined by light and scanning electron microscope to declare the morphology of the luminal epithelial surface. It was found that ciliated cells were present in one case. This finding explains the origin of ciliated cells as a development included in the neighbouring lymph node with a tendency of metaplastic change and exfoliation.     

A scanning and transmission electron microscopic study of experimental extrahepatic cholestasis in the rat

The rat liver after extrahepatic biliary obstruction was studied by SEM and TEM in correlation with basic histochemical techniques. Cholestasis was verified by serological methods. The biochemical data (increase in serum bilirubin values, a gradual lowering of the albumin fraction), in agreement with the ultrastructural results of a sparse RER, suggested a gradual decrease of the protein synthetic activity of the hepatocyte. SEM and TEM revealed numerous fat-storing cells, closely associated with patches of connective fibrils in the subendothelial spaces. Further ultrastructural observations demonstrated: a) a proliferation of the intrahepatic biliary tree (ductular proliferation, including newly formed ducts with sacculation and diverticuli); b) an increased number of canaliculo-ductular junctions and, c) an increase in the length of the bile canalicular network due to its tortuous course, pocketing and side branching. The occurrence of an intact cytoplasmic barrier separating the bile canalicular lumen from the Disse's space together with the results obtained by retrograde infusion of ferritin into the biliary tree suggested that the regurgitation pathway by ductular reabsorption and by transhepatocytic transport is the best documented and most acceptable, at least in the rat.     

Three-dimensional analysis of the vascular system in the rat spinal cord with scanning electron microscopy of vascular corrosion casts. Part 1: Normal spinal cord

Vascular corrosion casts of polyester resin in the normal spinal cord at C4-C6 and C7-T1 were inspected three-dimensionally by scanning electron microscopy in 13 rats. Arteries and veins were easily differentiated by the impression pattern of endothelial nuclei on the casts. The centrifugal arterial system from the sulcal arteries supplied most of the gray and white matter in the ventral and lateral spinal cord. Each sulcal artery supplied only one side of the cord. The average number of sulcal arteries was 2.6 per mm. The centripetal arterial system from the posterior spinal arteries fed the posterior gray and white matter. In contrast with classical concepts, there was no pial arterial plexus on the ventral and ventrolateral surface except for infrequent transverse branches from the anterior spinal artery. In the posterior columns, two types of large veins were identified: the posterior medial septal veins and the posterior oblique veins that drained the posterior columns, medial posterior gray matter, and posterior gray commissure. The remainder of the gray and white matter was drained by the sulcal veins and the radial veins. This method clearly demonstrates the three-dimensional structure of both the arterial and venous system in the rat spinal cord.     

Morphological variations of the human gastric mucosa after omeprazole treatment: a scanning electron microscopic study

The effects of antiepileptic drugs on cognitive function in 48 healthy volunteers were assessed using event-related potentials (ERP) and the Attention Index included in the Wechsler Memory Scale, revised edition (WMS-R). The study was conducted over 1 week, using a double-blind design. Four drugs, carbamazepine (CBZ), phenytoin (PHT), valproate (VPA) and zonisamide (ZNS) were tested. Using an auditory oddball task, ERP measurements were made under two conditions with different tone intensities: Condition 1 used 70 db SPL; and Condition 2 used 30 db SPL. Results showed that CBZ prolonged target N1 and P3 latencies in Condition 1, and reduced frequent N1 amplitude in Condition 2, which suggests that CBZ may cause a change in sensory memory and prolong stimulus evaluation time. It is suggested that under a low stimulus intensity level, the sensory function itself was affected. Phenytoin was found to prolong target N1 latency in Condition 2, which also indicates a change in the sensory memory function. However, VPA did not significantly affect ERP components, except for the shortened frequent N1 latency, which could not be explained due to the limited information. It was found that ZNS augmented P3 amplitude in Condition 2, and reduced scores on the Attention Index. It is suggested that the augmentation of P3 amplitude was caused by the reduction of processing negativity as a result of the detrimental effect of ZNS on subjects' attention. However, the apparent difference between the ERP and behavioral indices suggests that caution should be exercised in assessing the results obtained only from ERP measurements.     

An experimental scanning electron microscopic study of human cerebellar cortex using the t-butyl alcohol freeze-drying device

Specimen preparation methods are very important in scanning electron microscopy (SEM) of nerve tissues. In the present study, a t-butyl alcohol freeze-drying device was used to prepare cerebellar cortex of the human and that of the rat at 15 degrees C and 160 mm Hg. This method has been previously used with success in the preparation of other tissues, such as pancreas and trachea. Relatively large specimens (about 10 mm x 15 mm x 1 mm) of formalin-fixed human and glutaraldehyde-Millonig buffer perfused (1 hour) Wistar rat were rinsed in water, dehydrated in a series of ethanols, immersed in t-butyl alcohol, and then placed in the new freeze-drying device. The specimens were cut with a razor, freeze-dried without acid or alkali digestion, mounted on stubs, and sputter-coated with gold. This new preparation method allowed a higher magnification examination of surfaces of cells and fibers of the human cerebellar cortex compared to the critical point drying method. This was valid for Purkinje cell bodies with axons, dendrites with climbing fibers and climbing fiber glomeruli, and stellate neuron cell processes connected to the Purkinje cell dendrites. Lugaro cell, basket cells with axons, Golgi II cell, mossy fiber glomerulus with granule cell dendrites, satellite Bergmann glial cells with processes, and many microtubule-like fibrous structures on the inside of Purkinje cell dendrites were observed. Furthermore with this method, the glutaraldehyde-Millonig buffer perfused cerebellar cortex of the Wistar rat shows better three-dimensional images than the formalin-fixed human cerebellar cortex.     

A scanning electron microscopic study of the degenerative cartilage in patellar chondropathy

Patellar chondropathy as cartilage degeneration localized in patellar cartilage in young persons is characterized by cartilaginous changes, such as softening, swelling, and fissuring. With a view to structural characterization of early cartilaginous degeneration before erosion, the morphology of affected cartilage was studied under a scanning electron microscope. The surface network of cartilage constituting fibrils had an edematous change, presenting with fibrillation on the medial facet, whereas many fibrils of the central ridge had a collagen bundle, and fissuring of varying size was observed. It appeared that a mechanical force (shearing) acting on the site of the central ridge was associated with the formation of a collagen bundle and its destruction. On the lateral facet, fibrils were arranged perpendicular to the joint surface; the superficial layer of fibrils was worn by hyper-pressure acting on the lateral facet. On the fractured surface, the coarseness of collagen fibrils showed changes that varied with the site and stage of cartilage degeneration. Frequent changes were signs of fibril loosening (coarsening), such as reduction in fibril density (i.e., edematous change), collagen fibril aggregation, and fissuring, and longitudinal restructuring of fibrils. The patellar cartilage in the patients of this series showed a structure adapted to the mechanical force. The initial structural changes of cartilage consisted of collagen fibril aggregation and reduction in fibril density. These changes give rise to matrix rarefaction, which in turn causes cartilage degeneration to progress. These changes were concurrent in both the superficial and middle layers and were not localized as basal degeneration.     

Fluoride toxicity: a biochemical and scanning electron microscopic study of enamel surface of rabbit teeth

The present study has been carried out to investigate the effect of fluoride toxicity on the morphology as well as inorganic chemical constituents of rabbit teeth. Rabbits were administered sodium fluoride at a dose of 10 mg NaF/kg body weight every 24 h for 18 and 23 months. The incisor and molar teeth (whole tooth) were investigated for fluoride, calcium and phosphorus content in 18- and 23-month treated animals. The enamel surfaces of incisor teeth of 23-month treated animals were examined under scanning electron microscope. A significant increase in fluoride levels and significant decrease in calcium content was found following fluoride administration for 18 and 23 months as compared to control. Ca/P ratio was significantly increased only in 23-month treated animals. The scanning electron micrographs revealed hypoplastic, rough, uneven, pitted and cracked enamel surfaces covered with granular deposits as a result of excessive intake of fluoride. It can be concluded that long term fluoride administration leads to severe structural alterations on the enamel surface, possibly through defective mineralization.     

Cell membrane maturation of Friend erythroleukemic cells and tocopherol-dependent erythropoietin effect: a scanning electron microscopic study

Typical morphological features of surface structural alterations during Friend cells differentiation are described. Scanning electron microscopy (SEM) revealed that DMSO induction switched on cell alteration of the proerythroblast-like cells, possessing microvilli projections on cell membrane with some ruffles, to an advanced stage with a blebby surface. This was followed by the formation of a pear-like polarized cell separated into two zones by a narrow cytoplasmic bridge at the equatorial plane. The polarized cells showed a smooth surface and tended to disconnect into two unequal cells. The villous leukemic erythroblast has negatively charged sialic acid residues on the glycocalyx, available for latex hydrazide probe binding, while the blebby and polarized cells lack it. Tocopherol added to culture medium of DMSO-induced erythroleukemic cells prevented the formation of blebs and the polarization phenomena, without affecting hemoglobin synthesis. The tocopherol-treated cells contain available negative charges for latex hydrazide binding similar to uninduced Friend cells. Erythropoietin potentiated a repolarization ability and morphological alteration capacity to the tocopherol-treated cells and this was accompanied by a loss of glycocalyx-negative charges. At these growth conditions erythyropoietin induced a dose-dependent proliferation effect.     

Organization of the blood and lymphatic microvasculature of the gallbladder in the guinea pig: a scanning electron microscopic study

Fasciola hepatica adult worm cysteine proteinases were active-site, affinity radio-labeled with benzyloxicarbonyl-L-tyrosine-L-alanine diazomethylketone (Z-Tyr125I-Ala-CHN2). Sera from patients with fascioliasis and from rabbits experimentally infected with F. hepatica immunoprecipitated the radiolabeled parasite cysteine proteinases in immunoelectrophoresis assays. Two purified antigens were identified as part of the complex mosaic of antigens immunoprecipitated by the sera of infected patients. These antigens (Fas1 and Fas2) have been shown to be an important part of the Fharc2 precipitin band used for serologic diagnosis in humans and cattle. They showed cysteine proteinase activity with different proteolytic specificities and partial identity in double immunodiffusion assays. The results obtained in this work show that the Fas1 and Fas2 antigens are sensitive and specific antigens for diagnosis of this serious helminthic disease in humans and other susceptible hosts.     

Subclinical surface alterations of human pleura. A scanning electron microscopic study

Pleuritis or pleural effusion frequently develops in patients with pneumonia or heart failure. Most of these pleural changes regress without intrapleural intervention. The detailed mechanisms of the regression of the pleural changes in humans are not well documented. We studied the parietal pleura of nine patients with lung cancer and two patients with coronary artery disease by scanning electron microscopy (SEM). All patients had neither radiographic nor gross evidence of pleural disease but all had mixed surface alterations by SEM. Focal denudation of mesothelial cells was common. Deeper injuries exposed thick and thin interweaving collagen bundles. Patchy depositions of amorphous or crystallized fibrin covered normal and damaged pleural surfaces, frequently admixed with macrophages, red blood cells, and tissue debris. Reactive mesothelial cells appeared to proliferate over the fibrin. Our findings suggest that subclinical pleural alterations occur often in patients with pulmonary or cardiac diseases and that an intact pleural surface in those patients is restored mainly by the proliferation of reactive mesothelial cells.     

A scanning and transmission electron microscopic study on rat bone marrow sinuses and transmural migration of blood cells

Bone marrow sinuses of young rats were examined under the scanning (SEM) and transmission electron microscopes (TEM). Marrow sinus wall was composed of three layers: an inner or luminal endothelium, an outer or adventitial cell layer, and a basal lamina in between. The luminal surface of the endothelial cells was quite smooth and showed some fenestrations, which could be divided into two types according to their size. One was represented by larger fenestrations (1-3 mum in diameter) which were presumed to be formed transiently at the site of blood cell migration, while the other by small pores (0.1 mum) grouped into a cribriform area. The adventitial cells showed a discontinuous layer in the TEM. Under the SEM, the discontinuity corresponded to the spaces formed between the cytoplasmic attenuations of the cells. Blood cell migration from the extravascular hemopoietic tissue into the sinus lumen was numerously observed. The migration occurred not through an intercellular gap, but through the larger intracellular fenestration of the endothelial lining mentioned above. A number of megakaryocytes were identified by their bulky cytoplasm in the parenchyme. Figures suggesting the sequence of platelet liberation from this cell could be demonstrated. First, the megakaryocyte extended its peripheral cytoplasmic processes into the sinus through endothelial fenestrations. The processes, being conspicuously extended, became periodically constricted. Finally, platelets were believed to be produced by separation at the constricted portions and liberated to circulation. The occurrence of a few endothelial fenestrations apparently unassociated with blood cell migration may possibly be ascribed to detachment of a blood cells due to vascular perfusion. The functional significance of the adventitial cell was discussed in association with blood cell migration.     

A scanning and transmission electron microscopic study of fiber arrangement in the hepatic capsule

The arrangement of fibrous elements in the rat hepatic capsule was examined under a scanning electron microscope (SEM) after alkaline or acid maceration of the serous coat, in conjunction with examination of the thin sections using a transmission electron microscope (TEM). The elastic fibers appeared as thin threads in a densely meshed network, lying just beneath the serous coat. Their surface was granular with short rods in the materials fixed with paraformaldehyde. In contrast, the collagen fibers were observed as relatively thick threads, with fascicles of collagen fibrils that were uniform in size. These fascicles extended in various directions to form rough meshes that were traversed by small fascicles and anastomosed with each other. The fibrous branches of the elastic fibers extended on or into the underlying collagen fibers to be anchored, while the collagen fibers converged on many areas of the liver surface, and were transferred into the interlobular connective tissues. The findings of the present study thus suggest that the fiber arrangement plays an effective role in the mechanical protection of the fragile liver cells and delicate serous cells from pressure and friction damage by the neighboring abdominal organs and walls of the abdominal cavity due to the elastic mobility of the subserosal elastic network in addition to the possible slippery cushion of a serous layer on the serous cells.     

A combined scanning and transmission electron microscopic study and electron probe microanalysis of human pineal acervuli

The frequency of postoperative infections in oncologic head and neck surgery can be reduced by the prophylactic use of antibiotics. In order to assess such preventive treatments as to their advantages and disadvantages, a controlled clinical trial was undertaken. The prophylactic use of antibiotics was used in 107 patients, operated for tumors of the upper airway-digestive tract. According to a previous randomisation, the patients received either carbenicillin either ticarcillin. The efficiency of carbenicillin and ticarcillin proved similar. The results obtained with these antibiotics turned out to be superior to those previously obtained with combined ampicillin and cloxacillin; the number of wound infections, primary and secondary, was lessened. The most frequent complications were thrombophlebitis at the site of intravenous perfusion of the antibiotics and hypokaliemia.     

Angioarchitectural comparison of the filiform papillae of the cat and rabbit using scanning electron microscopic specimens

Women undergoing prenatal diagnosis (PND) for psychological reasons constitute a considerable proportion (10-20 per cent) of tests carried out on pregnant women. In these cases, the couple is below the lower limit for parental age, and there is no increased genetic risk. The aim of the present study was to describe the psychosocial background of women asking for PND on psychological grounds and to evaluate their wish for PND. A consecutive series of 51 women were interviewed in the 12th-15th week of gestation before attending for the test. Forty-five per cent were close to the limit for advanced maternal age (35-37 years). Most of them were educated and had stable partners; more than half of the women worked in nursing. Almost half (23/51) of them had previous psychic problems and 29/51 showed depressive mood. During childhood, 21 women had experienced severe disease or handicap. One-third (17/51) of them had been a consolidating member in their family. More than half (33/51) described considerable problems in relation to their mother or to motherhood and 10/51 in their relation to their father. Half of the women described at least three and only three women none of the following six identified predisposing factors: previous or present psychic insufficiency, experience of disability in childhood, role as a significant supporter in childhood, problems in relation to own mother, problems in relation to own father. These women's anxiety was understandable when their psychological history and current mental status were recognized. More than half of the women (29/51) were considered to have strong and 22 moderately strong psychological reasons for their desire to have prenatal diagnosis.     

Cerebral cortex blood flow and vascular smooth muscle contractility in a rat model of ischemia: a correlative laser Doppler flowmetric and scanning electron microscopic study

The present study was undertaken to ascertain the role of smooth muscles and pericytes in the microcirculation during hyperperfusion and hypoperfusion following ischemia in rats. Paired external carotids, the pterygopalatine branch of the internal carotids and the basilar artery were exposed and divided. Reversible inflatable occluders were placed around the common carotids. After 24 h, the unanesthetized rat underwent 10-min ischemia by inflating the occluders. Continuous cortical cerebral blood flow (c-CBF) was monitored by laser Doppler flowmetry. The measured c-CBF was below 20% of control (P < 0.001) during ischemia. A c-CBF of 227.5 +/- 54.1% (P < 0.001) was obtained during reperfusion hyperemia. A c-CBF of 59.7 +/- 8.8% (P < 0.001) occurred at the nadir of postischemic hypoperfusion, and this was followed by a second hyperemia. The cytoarchitecture of the vascular smooth muscles and pericytes was assessed by scanning electron microscopy. Samples were prepared using a KOH-collagenase digestion method. In control rats, arteriolar muscle cells showed smooth surfaces. Capillary pericytes were closely apposed to the endothelium. Immediately after reperfusion, transverse membrane creases were observed on the smooth muscle surfaces. During maximal hyperemia the creases disappeared. When c-CBF started to decrease the creases became visible again. Throughout the postischemic hypoperfusion the creases remained. Capillary endothelial walls became tortuous in the late phase of hypoperfusion. During the second hyperemia most arteriolar muscle cells showed smooth surfaces. Some pericytes appeared to have migrated from the vascular wall. The morphological changes of smooth muscle membranes suggest that they are related to specific perfusional disturbances during ischemia and reperfusion.     

Lipoblastic meningioma: a light and electron microscopic study

A patient with intracranial lipoblastic meningioma in the right frontal lobe is reported. The tumor was entirely made up of vacuolated cells. The nature of this rare neoplasm has been verified by light and electron microscopic studies.     

Microcirculatory patterns in adult rat cerebral hypophysis: a scanning electron microscope study of replicated specimens

The blood vascular bed of the cerebral hypophysis in the adult rat was replicated completely or incompletely by arterial injection of different amounts of methacrylate resin, to be observed with a scanning electron microscope. Complete replication confirmed our previous findings (Murakami et al., 1987) on the distribution and structure of the vascular beds in and around the hypophysis of the rat. One long major and several minor portal routes (vide infra) were reproduced sufficiently together with the systemic veins of the posterior lobe. Incomplete replication demonstrated that resin flows: 1) via the long portal vessels from the median eminence and neural stalk to the anterior lobe; 2) via the accessory long portal vessels from the subependyma to the anterior lobe; 3) via the short portal vessels from the posterior lobe to the anterior lobe; 4) via the neuro-intermedial portal vessels from the posterior lobe to the intermediate lobe; 5) via the intermedio-distal portal vessels from the intermediate lobe to the anterior lobe; and 6) via the tuberal portal vessels from the tuberal lobe to the anterior lobe. Incomplete replication also demonstrated that resin in the median eminence and neural stalk is drained preferentially into the anterior lobe via the long portal vessels, and that resin in the posterior lobe is drained mainly into the systemic veins. We were unable to demonstrate a retrograde resin flow from the anterior lobe to the median eminence, subependyma, neural stalk, intermediate lobe and posterior lobe, nor an ascending resin flow from the posterior lobe to the median eminence and subependyma. Also failing to be noted were an ascending resin flow from the hypophysis to the hypothalamus and a descending resin flow from hypothalamus to the hypophysis.