Light and electron microscopic observations on rat molar periodontal ligament after intraperitoneal injection of vinblastine

Suramin is a polyanionic compound with potent antineoplastic properties. Because polymorphonuclear leukocytes (PMNs) are a crucial component of host defenses against bacteria and fungi, the effects of suramin on PMN function were studied in vitro. PMNs from healthy donors were incubated with concentrations of suramin of 1 to 1,000 micrograms/ml (within and exceeding the therapeutic range) for 30 min, and PMN functional parameters were subsequently assessed. Suramin had no effect on viability, chemotaxis to N-formylmethionyl leucyl phenylalanine, phagocytosis of Candida albicans, or superoxide anion production in response to phorbol myristate acetate and formylmethionyl leucyl phenylalanine. Fungicidal activity against C. albicans blastoconidia was unaffected at a suramin concentration of < 500 micrograms/ml, whereas at higher concentrations a slight suppression was observed (P = 0.04). Bactericidal activity against Staphylococcus aureus was significantly suppressed by concentrations of > or = 100 micrograms/ml (P < 0.01). Phagocytosis of S. aureus was also significantly impaired at > or = 10 micrograms/ml (P < 0.05). The presence of 10% human serum during pretreatment did not abrogate the suramin-induced suppression of bactericidal activity. Treatment of PMNs with granulocyte colony-stimulating factor (4,000 U/ml) for 30 min prior to the addition of suramin (250 micrograms/ml) improved the bactericidal defect (P = 0.02). The PMN functional impairment may be related to increased susceptibility to bacterial infections, and granulocyte colony-stimulating factor may improve the defect.     

Light and electron microscopic findings in rhinosporidiosis after dapsone therapy

Morphological findings in serial nasal mucosal biopsies from three cases of rhinosporidiosis on dapsone therapy were compared with biopsies from 33 patients taken before dapsone or surgical treatment was initiated. All biopsies were examined by light microscopy and five by electron microscopy. Counts of histologically intact and degenerated organisms showed a decreasing proportion of intact forms with treatment.     

Laminin localization in enterocytic basement membrane of rat small bowel grafts. A light and electron microscopic study

In vitro laminins stimulate numerous biological effects, such as cell migration, proliferation, attachment and differentiation. In vitro laminins influence immunocompetent cells and in vivo possibly play an important role in graft rejection. To establish how laminins could be involved in the regulation of acute rejection of small bowel allografts (with and without immunosuppression), we investigated laminin distribution in rat small bowel allografts four days after transplantation, i.e., before the onset of histological signs of rejection, using antibodies against alpha1, beta1, gamma1 chain of laminin-1. In immunosuppressed allografts, the ultrastructure of the enterocytic basement membrane appeared normal, but no laminin staining was seen in this membrane, although basement membranes of intramural blood vessels and muscle cells were normally stained. In non-operated immunosuppressed rats, laminin staining was clearly reduced in the enterocytic basement membrane, demonstrating that cyclosporin A is able to affect this membrane. Since only rats in which laminin is altered survive, this laminin alteration in the enterocytic basement membrane presumably plays an important role in overcoming the acute rejection.     

Immunocytochemical localization of substance P receptor in rat periaqueductal gray matter: a light and electron microscopic study

The distribution of substance P receptor (SPR) protein in the rat periaqueductal gray matter (PAG) was investigated with a polyclonal antibody in the four subdivisions obtained by cytochrome-oxidase histochemistry (Co-hi). At light microscopic analysis, immunoreactivity appeared particularly dense in the dorsal subdivision of the PAG, was less intense in the other subdivisions, and formed several longitudinally organized columns. SPR-like immunoreactivity (SP(R-i)) was localized mostly to cell bodies and dendrites of small and medium-sized neurons, which constituted about 6% of the total neuronal population of the PAG. At the electron microscopic level, SP(R-i) could be observed on postsynaptic as well as on nonsynaptic regions of both cell bodies and dendrites. A small proportion of axons (4.2%) and axon terminals (5.3%) showed SP(R-i), the majority of labeled axon terminals, amounting to about 70% of synapsing elements, formed asymmetric synapses with dendrites. Rare astroglial processes displaying SP(R-i) were also observed scattered throughout the neuropil of all PAG subdivisions. Our observations suggest that 1) also in the PAG, SP may act in a diffuse, nonsynaptic manner, probably on targets that are distant from its sites of release; and 2) SP may modulate excitatory neurotransmission acting presynaptically on those labeled axons that form asymmetric synapses.     

Light and electron microscopic evidence for topographic and monosynaptic projections from neurons in the ventral medulla to noradrenergic dendrites in the rat locus coeruleus

Mental diseases such as schizophrenia are being modeled by artificial neural networks in an attempt to understand the underlying neuropathological processes. We studied hospitalized psychiatric patients that met the DSM-IIIR criteria for schizophrenia (N=19), and normal subjects with no psychiatric history (N=18). Performance on the Wisconsin Card Sorting Test (WCST) by schizophrenic patients was poorer than normal subjects as estimated by various scoring measurements. We then modeled an artificial neural network, motivated by biological considerations, that is able to simulate performance of normals and schizophrenics on the WCST. In order to model the complex nature of the WCST, we designed novel learning rules based on non-associative learning paradigms. We found that there must be a minimum amount of noise, or inherent synaptic instability, for our model to perform similar to schizophrenics.     

Intravital and electron microscopic observation of Ito cells in rat hepatic microcirculation

Intralobular distribution of Ito cells (fat-storing cells) in hepatic microcirculatory units was investigated through intravital fluorescence microscopy. By using a low-level ultraviolet epi-illumination and a sensitive silicon intensified target camera, the intrahepatic autofluorescence was visualized and digitally processed. In rats fed an ordinary diet, the ultraviolet-excited autofluorescence was composed of at least two different origins that could not be spectrophotometrically distinguished, namely, multiple patchy autofluorescence activities along the sinusoids and terminal hepatic venules and diffuse parenchymal autofluorescence. Multiple patchy activities showed a rapid photobleaching phenomenon under the continuous ultraviolet excitation. These fluorescent activities were completely eliminated by depleting the intrahepatic retinoid contents using a vitamin A-deficient diet for 4 weeks. Furthermore repeated administration of vitamin A significantly enhanced the patchy fluorescent activities. Electron microscopy revealed that these vitamin A fluorescent activities are colocalized with the fat droplets in Ito cells, providing evidence that Ito cells exist not only in perisinusoidal spaces but also in the perivascular spaces of the terminal hepatic and portal venules. The current intravital technique thus provides a new method to observe Ito cells in hepatic microcirculatory units in vivo.     

Quantitative autoradiographic light- and electron microscopic studies on the retinohypothalamic connections in the rat

Light microscopic autoradiography performed subsequent to intraocular injection of 3H-leucine revealed silver grains (SG) above axons of the optic tract which could be followed into the ventral and caudal portion of the suprachiasmatic nuclei (SCN) and above the contralateral anterior hypothalamic nucleus (AHN). By high resolution photometric measurement and computer processing the labelled areas were analysed, thus yielding statistical data of the relative grain distribution. The highest SG density was found in the ventrolateral part of both SCN (SCvl), confirming earlier reports concerning retinohypothalamic connections. That area exhibiting a cytoarchitecture different from the remaining nucleus was traversed, however, by numerous labelled axons. In the caudal part of both SCN a specific projection field of retinal fibres could be located. Here, almost no traversing fibres contribute to the rather circumscribed marked area. In the ventral part of the contralateral AHN, diffuse labelling well above background levels could be observed. Distinction between bypassing and terminating fibres within the SCvl could not be made using light microscopy. Analysis of SG distribution of the SCvl with electron microscopic autoradiography revealed a specific localization of SG within presynaptic terminals containing clear vesicles and pale mitochondria.     

Serial light and electron microscopic changes in renal glomeruli in mice fed griseofulvin

The acclimation of the clawed toad Xenopus laevis to hyperosmotic solutions of NaCl (balanced solution of sea salt), urea or mannitol was studied. The animals could not be acclimated to salt solutions more concentrated than 400 mosm.1-1. Urea was tolerated till 500 mmol.1-1. Plasma osmolality was always hyperosmotic to the environmental solution, but with diminished osmotic gradient at the highest tolerated solutions. Plasma urea concentration approached 90 mmol.1-1, similar in the three solutions of acclimation. Urine volume was very small under all conditions. Serum aldosterone and corticosterone did not differ significantly, although there was a slight tendency towards lower aldosterone in the NaCl solution. In vivo water uptake in tap water acclimated animals was very small, and was higher in the other groups. Only the salt- and urea-acclimated, but not the tap water and mannitol-acclimated groups responded with a clear increase following injection of oxytocin or theophylline. In vitro urea fluxes were similar and invariable in both directions under all conditions. No significant effect of theophylline was observed. Sodium transport measured by the short-circuit technique in vitro was lower in salt- and mannitol-acclimation conditions, and was stimulated significantly under all conditions in response to serosal oxytocin or theophylline. It is concluded that Xenopus laevis can osmoregulate at a limited range of external solutions. It is limited in the increase of its plasma urea concentration; the transport properties of the skin do not change very much upon acclimation, except for the hydroosmotic response to oxytocin.     

Light and electron microscopic study on the pineal complex of the coelacanth, Latimeria chalumnae Smith

The pineal complex of the coelacanth, Latimeria chalumnae was studied light and electron microscopically. It consists of two vesicles representing parapineal and pineal organs. Both occur intracranially and openly communicate with each other and the brain ventricle. The entire complex shows a striking photoreceptor morphology with sensory, ependymal and nerve cells. The last cell type is more abundant in the parapineal vesicle than in the pineal organ. The following ultrastructural details of the parapineal are noteworthy: 1. The sensory cells possess large inner and outer segments protruding freely in the vesicular cavity. The outer segments measure 8-10 mum in length and consist of as many as 275 lamellae. The basal processes of these cells terminate in neuropil-like regions. Occasionally, dense granules (500-1000 A) of uncertain identity occur in the perinuclear and inner segment cytoplasm of the cells. 2. The supporting cells are of the ependymal type. Their cytoplasm contains a filamentous feltwork and pinocytotic vesicles, but lacks secretory granules. Cytosomes are particularly abundant in cell processes in the neuropil-like zones. The basal end-feet of these cells isolate the receptor and nerve cells from the perivascular space. 3. In the neuropil-like regions, terminals of sensory cells make synaptic contacts with neuronal dendrites. Synaptic ribbon-like profiles in the terminals characterize the contact zones. Only unmyelinated nerve fibers could be observed in the small area of the tissue examined. The results are discussed with regard to photoreceptive and secretory functions of the pineal complex and its evolution in lower vertebrates.     

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.     

Light and electron microscopic analysis of liver sinusoids during hepatocarcinogenesis with 2-acetylaminofluorene in rats

To clarify the sequential changes and morphological differences of the sinusoidal structures between hepatocellular carcinoma (HCC) and hepatocellular adenoma (HA), we examined morphological changes of sinusoidal cells and related structures such as basement membrane during hepatocarcinogenesis in the rat. During continuous feeding of carcinogenic diets containing 2-acetylaminofluorene to rats, HA appeared at the 8th week in the peripheral area and then extended toward the centrolobular area. The appearance of HCC was recognized at the 27th week. In the HA lesion, the morphology of sinusoidal cells and related structures was basically the same as that of normal liver except for a slight thickening of the basement membrane and a decreased amount of vitamin A-lipid droplets of stellate cells. In HCC, the fenestrations of endothelial cells disappeared and the basement membrane became continuous, thick and often multilayered. Stellate cells contained almost no vitamin A-lipid droplets and were associated with abundant collagen fibers. Kupffer cells and pit cells were not seen inside the sinusoid. All these features of the sinusoids in HCC resembled the morphological characteristics of the capillary. The present study has revealed that HCC possesses sinusoid structures distinct from those of HA. This suggests that HCC may not derive directly from HA but may develop newly within the HA.     

Sarcomatoid renal carcinoma with angiosarcomatoid component. Light microscopic and immunohistochemical study

We describe a case of primary renal pelvic carcinoma which showed an unusual histologic pattern of anastomosing blood-filled channels lined by atypical cells. This angiosarcomatoid pattern merged with solid areas typical of renal carcinoma. Immunoperoxidase stains for cytokeratin and epithelial membrane antigen were positive in both the angiosarcomatoid and typical carcinomatous areas, while stains for factor VIII-related antigen and desmin were negative. Since primary renal angiosarcomas are rare neoplasms, the diagnosis of sarcomatoid renal carcinoma should be considered in primary renal neoplasms which show this angiosarcomatoid pattern.     

Evidence for stimulus access to taste cells and nerves during development: an electron microscopic study

We have examined developing taste buds in fungiform papillae of rats from the 18th day of gestation (E18) to the 15th postnatal day (P15). Nerve processes were seen in the epithelium of E18 rats before taste buds were obvious. At E20, early taste buds were visible, but were embedded within the epithelium, i.e., their cells were shielded from the oral cavity by overlying squamous epithelium. At this stage, the epithelium on the lateral aspects of the fungiform papillae was keratinized, but that overlying the taste bud was not. Some taste bud cells at E20 contained synapse-like structures near their contacts with nerve processes. In postnatal animals, keratinized epithelial cells were seen overlying taste buds, but taste pores were not observed until P10. How, then, do stimuli reach the taste cells and elicit physiological and behavioral responses as reported by others? The keratinized epithelium overlying the buds was unlike that on the lateral aspect of the papilla in at least one significant way. Few lamellated bodies were present in intercellular spaces beneath the stratum corneum, whereas these were abundant in the corresponding location within epithelium on the slope of the papilla. Although some were present within the squamous epithelium overlying the bud, they apparently were not released into the intercellular space. These lipid-rich lamellated bodies are thought to represent the water barrier of the epithelium, i.e., the barrier which prevents aqueous solutions from passing through the epithelium. We determined that the keratinized epithelium overlying the taste bud was permeable to a tracer, lanthanum nitrate, whereas that on the lateral surface was not. Lanthanum was visualized around taste cells and around nerve profiles within and near the taste bud. We propose that the absence of an aqueous permeability barrier in the epithelium overlying taste buds likely explains the ability of tastants to reach the taste bud cells and nerves in the developmental period before pore formation.     

Regional differences in the pineal gland of the cotton rat, Sigmodon hispidus: light microscopic, electron microscopic, and immunohistochemical observations

Light microscopic, electron microscopic and immunohistochemical observations of the various portions of the pineal gland of the cotton rat (Sigmodon hispidus) were made. The volume of the proximal half occupied about 30% of the whole organ, and pinealocytes were slightly smaller in size in the proximal portion than elsewhere. The distal and intermediate portions contained few interstitial cells and numerous astrocytes, but the proximal portion lacked interstitial cells and had more abundant astrocytes than elsewhere. Astrocytes, which were immunoreactive for glial fibrillary acidic protein, mainly lined the pericapillary spaces in the distal and intermediate portions, but in the proximal portion these cells often surrounded isolated or groups of pinealocytes. In the distal and intermediate portions, abundant sympathetic fibers and less numerous non-sympathetic, peptidergic fibers were mainly localized in the pericapillary spaces; these fibers were sparsely distributed in the parenchyma close to interstitial cells or astrocytes. In the proximal portion, non-sympathetic fibers were scarce and sympathetic fibers were distributed abundantly and almost exclusively in the parenchyma. Most of the sympathetic fibers were adjacent to astrocytes and, occasionally, made specialized contact with them. Fenestrae in the capillary endothelium were numerous in the distal portion but absent in the proximal portion. Thus, marked differences in structure existed between the distal and proximal portions of the pineal gland of the cotton rat suggesting that both portions are functionally dissimilar. In addition, the present study indicates that the proximal portion of the cotton rat was well developed and showed morphological features similar to the deeply situated pineal glands of other mammals.     

DOCA administration increases renal phospholipase activity in the rat

The phospholipase activity of renal tissue has been evaluated in controls and in DOCA treated rats. DOCA treated animal showed a higher than normal enzyme activity. Since a phospholipase is the key step in prostaglandin biosynthesis, it is suggested that the increased prostaglandin release promoted by mineraloactive steroids is mediated by an activation of this key enzyme.     

Aging rat vestibular ganglion: II. Quantitative electron microscopic evaluation

This laboratory has shown that age-related vestibular ganglion cell loss does not occur in the Wistar rat as it does in humans. However, in that study, intracellular changes were evident. The purpose of the present study was to quantitate some of these changes. The volume densities of mitochondria, rough endoplasmic reticulum (RER), Golgi apparatus, and aging pigment, as well as the diameter of the vestibular ganglion cells, of young (3 to 5 months) and old (24 to 31 months) female Wistar rats were determined by electron microscopy and stereological techniques. The data show a significant decrease in the volume densities of mitochondria (11.4%), Golgi apparatus (8.1%), and RER (8.9%), a significant increase in aging pigment (327%), and no change in mean profile diameter. These results suggest a decreased capacity for oxidative metabolism and protein synthesis that may reflect a decrease in the number of hair cells innervated by each ganglion cell and/or in the number of central connections. In either case, these findings suggest impaired metabolic and functional capabilities.