Reticular fibers in the vocal fold mucosa

Light and electron microscopic investigations of the reticular fibers (RFs) in the vocal fold mucosa were carried out on excised human adult larynges. The results are summarized as follows. Reticular fibers were found in the superficial and intermediate layers of the lamina propria of the vocal fold mucosa. They were most abundantly discovered around the vocal fold edge, and they decreased toward the superior and inferior portions of the vocal folds. The RFs were composed of slender fibrils, about 40 nm in diameter, and having cross-bands with a periodicity of about 67 nm. They were found in close association with the basal lamina of the epithelium and blood vessels. The slender fibrils of the RFs did not form any bundles, but branched and anastomosed. The RFs formed delicate 3-dimensional networks, and the spaces among the fibers were relatively large. Glycoprotein and glycosaminoglycan (proteoglycan) were situated around the RFs and in the spaces among the fibers. Elastic fibers were located in the spaces among the RFs. The 3-dimensional structure of the RFs in the vocal fold mucosa, first demonstrated in this study, appears to be one of the key components of the structural maintenance and viscoelasticity of the vibrating vocal fold tissue.     

Scanning electron microscopic view of auditory tonsil in turkey

The auditory tonsil of the turkey was studied by scanning electron microscopy. It is a sizable mass of lymphatic tissue dorsal to the infundibular opening in the auditory tube. The infundibular opening is in the roof of the turkey's mouth, caudal to the nasal cleft, and is the external orifice of the auditory tube that connects with the air spaces of the pneumonic bones in the head. Folds of the auditory tonsil that project toward or into the infundibular opening contained numerous lymphatic nodules in their lamina propria. The epithelial surface of the folds was covered with ciliated and nonciliated columnar cells, goblet cells with microvilli, and ductal openings of mucous glands located in the area. The lymphoid nodules were either oval or round and consisted of a thick reticular network that contained lymphocytes, fibroblasts, and erythrocytes. From some of the lymphatic nodules, there were sinusoids that contained lymphocytes and extended to the epithelial surface, whereas from others there was a lymphocytic infiltration of the surrounding lamina propria.     

Segi's cap: group formation of gut endocrine cells at the tip of the villi in human embryonal intestine

Large groups of basal-granulated (entero-endocrine) cells occur on the top of villi in the duodenum and upper jejunum of the human fetus older than five months of gestation. This structure was discovered and described by Segi(1935), hence named Segi's cap. By the immunocytochemical technique, somatostatin-, gastrin- and motilin-producing cells were demonstrated. By electron microscopy, EC, D and S cells were identified in the cap. There were autonomic nerves in the lamina propria adjacent to Segi's cap. No intraepithelial lymphocytes were found in the villi with Segi's cap.     

Structural organization of lymphatics in the monkey esophagus as revealed by enzyme-histochemistry

The fine structure and distribution of lymphatic vessels in the monkey esophageal wall were studied by light and electron microscopy using an enzyme-histochemical method. Identification of lymphatics was made by 5'-nucleotidase staining, whereas blood vessels were visualized by alkaline phosphatase staining. This technique revealed intramural lymphatic networks in the mucosa, submucosa, and myenteric layer of the esophagus. The organization of lymphatics in the esophagus basically conformed to that of the small intestine, although they showed certain distribution patterns peculiar to the esophagus. The lamina propria mucosae exhibited a double-layered lymphatic network, and lymphatic capillaries extended into its papillae. Despite their forming blind ends and being closed by endothelial cells, the lymphatics in the mucosal papillae were found to contain lymphocytes in their lumen. This suggests that free cells might penetrate the endothelium to enter these initial portions of the lymphatics.     

Ultrastructural and immunohistochemical studies of intralobular fibroblasts in human submandibular gland: the recognition of a 'CD34 positive reticular network' connected by gap junctions

In order to clarify the ultrastructural and immunophenotypic characteristics of intralobular stromal cells in the human submandibular gland, normal tissues were examined from 25 patients by conventional histology of wax sections, light microscope immunohistochemistry, conventional thin-sectioning transmission electron microscopy, immuno-electronmicroscopy (for the CD34 antigen) and scanning electron microscopy. Spindled stromal cells had abundant rough endoplasmic reticulum cisternae typical of fibroblasts. Some actin filaments and microtubules were seen, but the filaments lacked the focal densities of smooth-muscle, nor was a lamina present. Fibroblasts exhibited long slender processes which associated with other fibroblasts by means of gap junctions and with mononuclear cells (lymphocytes, plasma cells, macrophages) by means of close appositions of membrane. Immunoelectron microscopy for CD34 revealed positive staining over the fibroblast surfaces but not over the mononuclear cells. The novel observation of gap junctions suggests a tissue closely organized as a network in physical and perhaps physiological terms. This network also incorporates mononuclear cells. The possible role in immunosurveillance of this CD34-positive reticular network is discussed.     

Parenteral versus enteral nutrition: morphological changes in human adult intestinal mucosa

In animal experiments total parenteral nutrition induces an atrophy of the small intestinal mucosa. In humans morphological data are few and controversial. Therefore, the aim of this study was to investigate the effect of parenteral nutrition on the intestinal mucosa of human adults. For this purpose samples of the proximal jejunum of a) patients with chronic pancreatitis receiving total parenteral nutrition as presurgical treatment, b) enterally nourished patients without (controls) and c) with chronic pancreatitis were compared using light and scanning electron microscopy. Statistical differences were assessed applying computer-assisted morphometry. The results demonstrated that the thickness of the jejunal mucosa decreased already in enterally nourished patients with chronic pancreatitis. However, after total parenteral nutrition the decrease (atrophy) was enhanced due to a strong reduction in villus height albeit the crypt length increased. In addition, scanning electron microscopy revealed distinctive changes in mucosal surface pattern, whereby finger-like villi were replaced by leaf-like villi and by long, winding bifurcating ridges. Cell shedding was absent. In conclusion, total parenteral nutrition in humans induces 1) an atrophy and 2) a remodelling of the intestinal mucosa (epithelium and lamina propria) with a decrease in the absorbing surface. These alterations involve both cell proliferation and cell shedding. The response of the mucosa to parenteral nutrition is immediate and the effect of the treatment in bringing about morphological alterations is more efficacious at the beginning than in the successive period. The basic disorder (chronic pancreatitis) of the patients nourished parenterally contributes to mucosal atrophy, but not to remodelling.     

Possible significance of cells within intraluminal collagen masses in equine oviducts

In addition to the unique feature of retention of unfertilized ova, the oviducts of mares frequently contain large intraluminal masses with a fibrillar component and some cells. The aim of this study was to identify the cells and examine their relationship to the extracellular components of these masses. Intraluminal masses were examined both in situ and flushed from the oviducts. The nature of the contained cells and their relationship to the fibrils were examined by light microscopy and by transmission and scanning electron microscopy. In some mares the large masses distended the oviduct, but neither loss of the oviductal epithelium nor damage to this epithelium was seen. Electron microscopy verified that the principal cellular component was fibroblasts, and that the fibrils were type I collagen. Collagen masses collected shortly after ovulation frequently contained viable fibroblasts with collagen fibrils associated with their cell surfaces and with surface clefts. Although such collagen masses were present in pregnant and nonpregnant mares, masses with viable fibroblasts were chronologically associated with recent ovulation. It was concluded that connective tissue drawn into the oviduct at ovulation is retained, and collagen synthesis continues at least for a few days. Although the fibroblasts eventually disintegrate, the collagen remains and may in some cases aggregate within the oviductal lumen to the extent that oviductal transport and embryonic viability could be affected.     

The localisation of particles retained in the trachea of the rat

Particles of 133BaSO4 were deposited on the surface of rat trachea by intra-tracheal injection. Aggregates of particles were located in the trachea by autoradiography and electron microscopy. Following deposition, particles not rapidly removed by muco-ciliary clearance remained on the epithelium for some time. After 2 hr most had been ingested by macrophages on the surface, though some were still free in the mucus. By 24 hr, 74 per cent. of the aggregates remaining were beneath the epithelium in the lamina propria, and after 7 days almost all of them were in or beneath the epithelium. All the buried particles identified by EM were within macrophages. After 24 hr the particles in the tracheal wall were beneath epithelium which was not ciliated columnar, but cuboidal or flatter, with fewer or no ciliated cells and infiltrated with lymphocytes. It is suggested that particle retention in airways is accomplished by ingestion by macrophages which then migrate through this type of epithelium.     

B and T lymphocytes are enhanced in the gut of piglets fed heat-treated soyabean proteins

Gut immune responses have been suspected in food hypersensitivity reactions such as those to soyabean proteins in early-weaned piglets. The present study examines the lymphoid cell subset distribution in piglets fed heat-treated (HTSP) or ethanol-treated soyabean proteins (ETSP). Duodenal cryosections of 4-week-old HTSP piglets (n = 10) and ETSP piglets (n = 8) were analysed for IgA, IgM, IgG1 and IgG2 positive cells, CD2, CD4, CD8, WC1 T cell positive antigens using immunohistochemical peroxidase reactions. Densities of IgM+ and IgA+ cells were three times and, IgG1+ and IgG2+ six times higher in the lamina propria of HTSP piglets compared with ETSP (P < 0.05). Increased CD2+ T cells were accounted for by a rise in both CD4+ and CD8+ T cell subsets in the lamina propria (P < 0.01) as well as in the epithelium of the duodenal mucosa of piglets fed HTSP. The density of the WC1+ T cell subset in the epithelium was significantly higher in HTSP than in ETSP piglets (P < 0.01). Immune reactions in the duodenal mucosa, involving both B and T lymphocytes may be related to atrophy of the duodenal villi in HTSP piglets.     

Fibroblasts are required for Schwann cell basal lamina deposition and ensheathment of unmyelinated sympathetic neurites in culture

The ability to purify and recombine populations of peripheral neurons, Schwann cells and fibroblasts in tissue culture has enabled us to examine the contribution of fibroblasts to Schwann cell basal lamina assembly and ensheathment of unmyelinated rat superior cervical ganglion neurites in vitro. Purified perinatal superior cervical ganglion neurons were grown in culture either with Schwann cells or with Schwann cells plus fibroblasts derived from either superior cervical ganglion capsule or cranial periosteum. The cultures were maintained for 2-8 weeks on a collagen substratum in a medium known to promote Schwann cell differentiation (myelin, basal lamina formation) in the presence of dorsal root ganglion neurons. The extent of Schwann cell differentiation (ensheathment, basal lamina formation) in the presence of superior cervical ganglion neurons was evaluated in this study using electron microscopy. In superior cervical ganglion neuron plus Schwann cell cultures (without fibroblasts), Schwann cells achieved only a moderate degree of ensheathment; also, Schwann cell basal lamina was discontinuous and extracellular collagen fibrils were sparse. Although only discontinuous basal lamina was demonstrable by electron microscopy in these cultures, surprisingly, Schwann cell/neurite fascicles were uniformly immunostained for laminin, type IV collagen, and heparan sulfate proteoglycan. The addition of fibroblasts to superior cervical ganglion neuron plus Schwann cell cultures increased the deposition of basal lamina around the Schwann cell/neurite units, the number of collagen fibrils, and the extent of neurite ensheathment. We propose that the presence of basal lamina increases the Schwann cell's ability to ensheathe superior cervical ganglion neurites, possibly through an augmentation of specific extracellular matrix components or by increasing in some way the capacity of these components to become organized into basal lamina. We conclude that, unlike dorsal root ganglion neurons, superior cervical ganglion neurons are unable to stimulate full Schwann cell extracellular matrix expression with the result that these Schwann cells require the extraneuronal influence of fibroblasts to deposit basal lamina and attain their mature phenotype in culture.     

Microsphere uptake by the intestine of White Leghorn chickens

A study was conducted to determine the effective size for latex microsphere uptake in the intestine of white leghorn chickens. Three trials were conducted in which ligated intestinal segments of anesthetized 8-wk-old chickens were injected with 0.2-, 0.5-, 2-, 6-, 10-, or 20-mu diameter fluoresceinated latex microspheres. Microspheres were counted in brush border, epithelium, and lamina propria of each intestinal segment, liver, and spleen. After 1 hr, the 0.2-, 0.5-, and 2-mu microspheres were oriented along the brush border of epithelial cells and microsphere uptake into the epithelium and lamina propria was observed in the duodenum, ileum, cecum, cecal tonsil, and colon. Uptake of microspheres of 6, 10, and 20 mu diameter into epithelium and lamina propria was not observed in any intestinal segment. Also, no microspheres of any diameter were observed in sections of liver and spleen to suggest that there was no appreciable entry of microspheres into the bloodstream within 1 hr after administration. The results indicated that uptake of microspheres by the chicken intestine is a size-dependent process with microspheres < or = 2 mu being taken up to an equal extent by most segments of intestine.     

Scanning electronic microscopy of the small intestine in persistent diarrhea

Persistent diarrhea very often leads children to malnutrition. It has become the major cause of death resulting from acute diarrhea episodes in developing countries. In order to determine the ultrastructural alterations of the small bowel that occur in the syndrome, 16 infants with severe persistent diarrhea were studied, utilizing light microscopy and the scanning electron microscope. Stool and jejunal fluid samples were collected for culture, rotavirus, ova and parasite search. Enteropathogenic agents were isolated in stools from 11 (68.7%) patients and bacterial proliferation in the small bowel was detected in 11 (68.7%) patients. EPEC strains were the most frequent enteropathogenic agent isolated both from stool and jejunal fluid cultures. The stool cultures revealed the presence of the following enteropathogenic microorganisms: EPEC 0111 in four, EPEC 0119 in one, EAggEC in five, Shigella flexneri in two, and Shigella sonnei in one; mixed infections due to EAggEC associated with EPEC 0111 were seen in two patients. The light microscopic analysis revealed that 56.2% of the patients suffered moderate villous atrophy most frequently associated with effacement of the microvilli, intracytoplasmatic vacuolization, increased number of multivesicular bodied and increased lymphocytic and eosinophylic infiltration in the lamina propria. The scanning electron microscopic analysis revealed in all cases shortening of the villi and enterocyte derangements; very often there was a total lack and/or effacement of the microvilli; in half of the patients there was a mucoid material covering the enterocytes tightly adhered to the apical epithelium surface. The scanning ultrastructural alterations observed in these patients are probably due to an association of factors brought about by the presence of enteropathogenic microorganisms and the resulting food intolerance that is responsible for perpetuation of diarrhea.     

Production of tubular structures from the aerial mycelium of Actinomyces roseoflavus var. roseofungini]

Jejunal lamina propria plasma cells and eosinophils and intraepithelial lymphocytes were raised in coeliac children on gluten-containing diets, but only intraepithelial lymphocytes were increased in patients on gluten-free diets. In contrast, lamina propria lymphocytes were reduced in children with coeliad disease on gluten-containing diets but were normal in paitents on gluten-free diets. In children with coeliac disease who were studied serially, lamina propria plasma cells and eosinophils and intraepithelial lymphocytes increased, and lamina propria lymphocytes decreased, within three months of the reintroduction of gluten to the diet. These observations are essentially similar to those made in the adult form of the disease and suggest that more than one type of immunological reaction is involved in the pathogenesis of the jejunal lesion.     

Low-risk differentiated thyroid cancer: the need for selective treatment

By means of an electron microscopic study of the intestine in young rats infected with Cryptosporidium parvum we observed a mass migration of immunocompetent cells of the host (eosinophils, neutrophils and macrophages) into the lumen of intestine. Some lymphocytes were also observed. Immunocompetent cells (except lymphocytes) included inside phagosomes with different endogenic states of C. parvum. Macrophages with typical extracytoplasmic parasitophorous vacuoles formed by C. parvum were also observed in the intestine lumen. Almost all stages of C. parvum could be observed on a surface of such macrophages. However, we did not find in lamina propria of intestine villi any macrophages with parasites. The place of macrophages infection is unknown. We suggest that surviving of C. parvum in macrophages is principally possible.     

Phenotypic distribution of T cells in human nasal mucosa differs from that in the gut

Phenotypic and functional studies are required to understand the immunoregulatory role of mucosal T cells. Information about T cells in the human upper respiratory tract is limited and conflicting. Therefore, we phenotyped T cells in nasal mucosa by means of multicolor in situ immunofluorescence. In normal mucosa, most CD3+ intraepithelial lymphocytes (IELs) and lamina propria lymphocytes (LPLs) (> 90%) expressed T-cell receptor (TCR)alpha/beta, and only approximately 5% expressed TCRgamma/delta. Although most IELs in the surface epithelium were CD8+ (64%), many expressed CD4 (30%) and the CD4 phenotype dominated (55%) only slightly in the lamina propria. This result was strikingly different from that obtained for comparable compartments in histologically normal jejunal mucosa, where IELs consisted of 83% CD8+ and LPLs of 73% CD4(+) T cells. Nasal CD3+ IELs and LPLs were mainly CD45RO+CD45RA- and usually expressed CD7. The integrin alphaEbeta7 was, as expected, more common on IELs than on LPLs (78 versus 20%). In conclusion, nasal T cells show several similarities to those of the normal jejunum but some notable differences exist, especially a relative increase in CD4+ T cells in the epithelium and a decrease in the lamina propria. It should be explored whether this disparity, together with an increased expression of epithelial adhesion molecules, might contribute to local immunological overstimulation and partly explain the relatively high frequency of airway allergy.     

Cutaneous lymphocyte associated antigen (CLA) and alpha e beta 7 integrins are expressed by mononuclear cells in skin and oral lichen planus

The cutaneous lymphocyte associated- (CLA-) positive subset of lymphocytes appears to migrate preferentially into skin by interacting with E-selectin on vascular endothelium; lymphocytes expressing the alpha e beta 7 integrin accumulate preferentially in the epithelium of the gastrointestinal tract. To determine whether the mononuclear cell population of the oral mucosa resembles that of the skin or intestine, and using lichen planus as a model, the proportions of CLA- and alpha e beta 7-positive cells in the epithelium, lamina propria and peripheral blood were compared by immunostaining and flow cytometry. In both skin and oral lichen planus, selective accumulation of CLA-positive cells was seen in the epithelium but not in the lamina propria. In contrast, large numbers of alpha e beta 7-positive intraepithelial cells were found in oral but not in skin lichen planus. These results show that in terms of CLA and alpha e beta 7 expression there are important differences in the mononuclear cell population of oral mucosa and skin.     

Gastric cryptosporidiosis in a wild frilled lizard from Australia

An adult male frilled lizard (Chlamydosaurus kingi) was found dead in Northern Territory (Australia). On physical examination it was found to be severely emaciated. At necropsy the stomach was found to be firm with mild thickening of the gastric mucosa. Gastric rugae were prominent and the mucosa was red with a thin layer of dark red mucus on its surface. Light microscopic examination revealed a mild diffuse gastritis with infiltration of the lamina propria by lymphocytes, plasma cells, and heterophils. Large numbers of small, round amphophilic to basophilic (2 to 4 microns in diameter) organisms morphologically consistent with cryptosporidia were seen on the surface of the mucosal epithelium and free in the gastric lumen. The gastric lesions seen in this frilled lizard did not involve atrophy, as previously described in lizards with gastric cryptosporidiosis, and were similar to those described in snakes. The possibility that more than one species of Cryptosporidium parasitizes reptiles could explain the different lesions. This is the first report of cryptosporidiosis in a frilled lizard.     

Enteric coccidia (Apicomplexa) in the small intestine of the northern spotted owl (Strix occidentalis caurina)

Sporulated oocysts (mean dimensions = 13.0 x 10.8 microns) and sporocysts (11.3 x 5.5 microns) of a coccidian resembling Frenkelia sp. or Sarcocystis sp. were present in the lamina propria of the small intestine of a naturally-infected northern spotted owl (Strix occidentalis caurina) collected near Medford, Oregon (USA). Dimensions of these oocytes and sporocysts appear to be considerably smaller than those from other sarcocystid species with avian definitive hosts. Additionally, numerous developmental stages and unsporulated oocysts (mean dimensions 22.8 x 17.8 microns) of a possible species of Isospora also were observed in the intestinal epithelium. This constitutes the first report of enteric coccidia from spotted owls. Neither parasite appeared to cause the death of the bird.     

Identification and characterization of spodoptera frugiperda furin: a thermostable subtilisin-like endopeptidase

Colonic histopathology in some patients with untreated celiac sprue and refractory sprue has been said to be indistinguishable from lymphocytic colitis, but there have been no objective comparisons on which this is based. The purpose of this study was to determine the prevalence and to characterize the nature of colonic histopathology at the time of diagnosis in patients with celiac or refractory sprue. Colonoscopic biopsy specimens obtained at the time of diagnosis from 16 patients with celiac sprue, six patients with refractory sprue, nine patients with lymphocytic colitis, and five normal controls were analyzed blindly by histological and morphometric methods, quantitating the number and specific subtypes of inflammatory cells within the lamina propria and epithelium. Immunoperoxidase staining of intraepithelial lymphocytes with a monoclonal antibody to CD8 also was performed. Three of 16 patients with untreated celiac sprue (19%) were thought to have colonic histological abnormalities, which by morphometry consisted of slightly increased numbers of lymphocytes in the surface epithelium and lamina propria, many of which were CD8-positive. These abnormalities were distinguishable from lymphocytic colitis by the lack of increased overall lamina propria cellularity and surface epithelial abnormalities, and by fewer intraepithelial lymphocytes. In refractory sprue, colonic histological abnormalities were more frequent than in celiac sprue, occurring in four of six patients (67%), more pronounced, and identical to those in the lymphocytic colitis syndrome. However, colonic intraepithelial lymphocytes in lymphocytic colitis were mostly CD8-positive, whereas those in the colitis of refractory sprue rarely were. Mild colonic lymphocytosis in patients with untreated celiac sprue should be distinguished from lymphocytic colitis by the lack of surface epithelial abnormalities, the lack of increased cellularity of the lamina propria, and the lack of ongoing watery diarrhea after treatment with a gluten-free diet. In contrast, colonic histopathology in refractory sprue is indistinguishable from lymphocytic colitis, although immunohistochemical differences do exist.     

Microvasculature of the dorsum of the rat tongue: a scanning electron microscopic study using corrosion casts

The purpose of this study was to examine the microvascular system of the dorsum of the rat tongue in relation to the subepithelial connective tissue relief. Methylmethacrylate corrosion casts of the vasculature as well as specimens in which the epithelium had been removed were prepared and examined by scanning electron microscopy (SEM). The SEM images revealed four distinct layers in the vasculature of the lamina propria. Superficially, Layer 1 comprised the irregularly shaped ends of capillary loops. Layer 2 consisted of parallel ascending and descending capillaries. These two layers were drained by the subjacent Layer 3, a fish-net-like venous plexus. More deeply, Layer 4 contained arterioles ascending from the muscular layers of the tongue. The vessels of larger papillary structures formed cylindric networks supplied by several arterioles and drained through a single venule to the venous plexus in Layer 3. At their base, the capillary loops often showed a constriction, suggestive of a sphincter. Arteriovenous anastomoses were rare. The epithelium-connective tissue interface showed short conical papillae as well as larger papillary structures which corresponded in both distribution and size to the capillary loops found in the vascular casts. The results confirm that the tongue is highly vascularized and demonstrate that the vascular supply to the papillae, taste buds, and other surface structures forms a highly organized and specific system which is adapted to the specific functions of this organ in the rat.