Rat parathyroid gland, with special reference to its blood vascular bed, pericapillary space and intercellular space

The blood vascular bed, perivascular space and intercellular space of the rat parathyroid gland were studied using scanning electron microscopy of vascular casts, freeze-cracked tissue samples, and NaOH-digested tissue blocks. The findings were supplemented by transmission light and electron microscopy of iron colloid-treated or enzyme-digested tissue sections. The rat parathyroid gland contained a rich network of capillaries. These capillaries were surrounded by marked pericapillary spaces which were demarcated by basal lamina of both capillaries and parenchymal cells. The pericapillary spaces contained numerous collagen fibrils, and issued many crista-like projections which ran deep into the sheets of parenchymal cells. The intercellular spaces of parenchymal cells contained neither basal lamina nor collagen fibrils. The surfaces of the parenchymal cells showed strong negative charging, and maintained the intercellular spaces. The luminal surfaces of the capillary endothelium also showed strong negative charging, and maintained the capillary lumen.     

Depletion of acinar secretory granules in the ferret parotid gland: effects of substance P and vasoactive intestinal peptide

Intercellular transport-metabolic interactions were investigated at the ultrastructural level in the grafts of the embryonic rat hippocampus and septum developing for 3 to 12 months in the anterior eye chamber of adult rats. The signs of highly increased transport from the blood capillaries ingrowing from the host's iris into the grafts (multiple pinocytotic vesicles in the endothelium and pericytes) were observed. The glial cells, which were accumulated at the graft surface, had pinocytotic invaginations and microvilli, which indicated their possible participation in the active transport of metabolites from the surrounding intraocular fluid. An increased level of direct communications, manifested in pinocytoses and large gap junctions between apposed nerve and glial cells was also present within the grafts. Moreover, microphagocytosis, as internalization of surface membrane fragments with adjacent cytoplasm of the neighbouring structure (including dendrites and axons), was often observed in the grafted tissue. It is suggested that the observed communications between neuronal and glial cells may participate in both trophic and functional interactions. An increase in the level of nonsynaptic interactions in the grafted nervous tissue may be regarded as a manifestation of the compensatory adaptation to the absence of normal tissue surrounding afferent connections and efferent targets.     

Diet-induced changes in the fatty acid composition of mouse hepatocyte plasma membranes

Hepatocyte plasma membranes were isolated from the livers of mice fed either a low fat diet or high fat diets containing polyunsaturated or saturated fat. The combined rate and isopycnic ultracentrifugation technique which was used produced highly purified hepatocyte plasma membrane fractions. The efficacy of the procedure was checked by electron microscopy and the assay to marker enzymes for the different subcellular organelles. Mice were maintained on a low fat diet until 60-70 days of age, when they were fed high fat diets containing polyunsaturated fat. The hepatocyte plasma membrane lipids of mice fed the polyunsaturated fat diet for 4 wk contained increased proportions of the major dietary unsaturated fatty acid, linoleic acid, and increased proportions of arachidonic acid. The proportion of linoleic and arachidonic acids decreased with continued feeding of the polyunsaturated fat diet. The hepatocyte plasma membrane lipids of mice fed the saturated fat diet contained increased proportions of oleic acid.     

Osteopontin expression in mammary gland development and tumorigenesis

Osteopontin (OPN) is a secreted phosphoprotein that binds to cells via an Arg-Gly-Asp sequence and to mineralized surfaces. The protein can mediate cell adhesion and is strongly implicated in transformation and tumorigenesis. We have examined the expression pattern of OPN in mouse mammary glands at different stages of postnatal development. Whereas OPN is expressed at low-to-moderate levels in mammary glands from virgin and pregnant mice, the levels of OPN mRNA are extremely high in the lactating gland, consistent with the presence of the protein in milk. Expression is highest at 2 days of lactation and declines thereafter, but it remains high through involution. OPN expression is restricted to small nests or groups of cells at 9 days of involution. These results suggest that OPN may play a specific role in the process of involution that may be distinct from its role during lactation. In mammary tumors arising spontaneously in transgenic mice expressing the oncogenes c-myc and/or v-Ha-ras under the control of the mouse mammary tumor virus promoter, the level of OPN expression is increased dramatically over that in the normal gland in these same animals. Numerous cells expressing OPN mRNA are widespread throughout the tumors. OPN protein is detectable by Western blotting in extracts from the mammary gland at 2 days of lactation and from the tumors, but not in mammary glands at other stages of development. We hypothesize that OPN is exported from most tissues and that the protein is only detectable in tissues elaborating fluids, such as the lactating mammary gland, or in pathological situations when expression of OPN is abnormally high, such as in tumors.     

Possible function of human brown adipose tissue as suggested by observation on perirenal brown fats from necropsy cases of variable age groups

In 134 out of 180 perirenal fat samples (74%) derived from Japanese necropsy cases aged from 1 month to 86 years, the brown fat tissue persisted in variable amounts. Brown fat cells were classified into 6 types: Type 1 cells are fat-depleted cells filled with granular cytoplasm and are believed to be produced after oxidation of fat for heat production. Type 2 cells are small-locular cells suitable for rapid oxidation of fat droplets. Type 3 (middle-locular) and 4 (large-locular) represent fat-storage cells containing large amounts of fat. Type 5 cells are thought to be transitional forms between multilocular brown fat cells and monolocular white fat cells. Type 6 (cytoplasm-rich multilocular) cells, usually found together with Type 1 cells, are thought to be fat-depleting or -consuming cells, since in them fat droplets are reduced in number and size probably in consequence of oxidation of fat, but by contrast granular cytoplasm is increased in amount separating the individual fat droplets by thick cytoplasmic septa. The occurrence of Types 1 and/or 6 cells that has been revealed in 65 out of the total 180 samples (36%), suggests that the oxidation of fat for the thermogenesis proceeds in the brown fat tissue and that brown fat cells partially undergo fat depletion. In the present study, the thermogenesis of human brown fat tissue was suggested chiefly with regard to the occurrence of Types 1 and/or 6 cells. In the majority of perirenal fat samples from infants (1-11 months) relatively numerous Types 1 and 6 cells were frequently revealed together with Type 2 cells, suggesting rapid and active heat production in support of the view that in human infants the brown fat tissue may be thermogenetically active to maintain body temperature. In the same manner, marked ability to produce a considerable amount of heat was evidenced in brown fat tissue of children and teenagers. In younger and elderly adults the frequency of occurrence and the amount of the perirenal brown fat tissue were decreased but Types 1 and/or 6 cells could be found in 17-40% of them, infrequently together, with Type 2 cells, suggesting persistence of the thermogenic activity with occasional large heat production especially in younger adults (20-39 years). Thus, the results obtained in this study have clarified that the human brown fat tissue can respond to stimuli given to the body by oxidation of stored fat even in the latest decades of life. In cases of death from burning, drowning, bleeding, drug poisoning etc., numerous Types 1 and/or 6 cells were found, suggesting that an active fat oxidation would take place in brown fat tissue assumedly as the result of the raised noradrenalin level in this tissue. The so-called small cytoplasmic cells found in perirenal fats from cases of death from liver cirrhosis and other causes were assumed to be atrophic fat-depleted brown fat cells.     

Influence of furosemide on the ureteric damage in a rat model of obstructive uropathy

The pineal gland of rats of various ages (1-21 days old) was examined by immunohistochemistry and electron microscopy. Numerous widely distributed cells identified as macrophages/microglia were immunoreactive with the monoclonal antibodies OX-42, OX-18, OX-6, and ED1, indicating that they expressed complement type 3 (CR3) receptors, major histocompatibility complex class I and II antigens, and antigens of monocyte/macrophage lineage as detected by the antibodies, respectively. Following an intraperitoneal injection of rhodamine isothiocyanate (RhIC) in all age groups, the cells emitted a bright fluorescence. They were also labeled by horseradish peroxidase (HRP), as demonstrated in both light and electron microscopy. An HRP reaction was observed in vesicles and lysosomes at the ultrastructural level. A remarkable feature was the uptake of these tracers by pinealocytes. In light microscopy, the pinealocytes showed a punctate reaction product 3-24 hours after HRP injection. By electron microscopy, the reaction product was observed in vesicles, lysosomes, and some rod-like structures in the cytoplasm. On the basis of their immunophenotypic features, it is suggested that the macrophages/microglia in the pineal gland are active phagocytes which are also probably involved in the immunoregulatory function in the gland. The avid uptake of RhIC and HRP from the circulation by these cells suggests that serum-derived substances that may gain access to the parenchyma of the gland are being constantly monitored. The labeling of pinealocytes with HRP suggests that the functional activities of these cells are being modulated by serum-derived substances.     

Alterations of lung structure in experimental diabetes, and diabetes associated with hyperlipidaemia in hamsters

Since hyperglycaemia is known to affect normal pulmonary physiology and biochemistry and few structure-function correlations have been reported, we designed experiments on hamsters subjected to streptozotocin-induced diabetes or diabetes associated with hyperlipidaemia, and investigated the impact of these conditions on the lung structure. At time intervals ranging 2-24 weeks from the inception of disease (without correcting blood glucose with insulin), the animals were sacrificed, and plasma glucose and cholesterol assayed. The lung was processed for electron microscopy, and the structural changes of the capillary and venular endothelium, of epithelial cells, and interstitium were examined. In diabetic animals, especially after 6 weeks of disease, a gradual narrowing of approximately 35% of the capillaries and approximately 30% of the alveoli, and hyperplasia of the extracellular matrix, rich in collagen bundles, were observed. Frequently, capillaries contained adherent intravascular macrophages suggestive of an inflammatory process. The capillary endothelium was characterized by numerous plasmalemmal vesicles, often fused, well-developed synthesizing apparatus (endoplasmic reticulum and Golgi complex) and cytoskeleton, and an uneven distribution of the anionic sites on the luminal plasmalemma. The venular endothelium was particularly rich in Weibel-Palade bodies. The alveolar epithelium was often collapsed, compressing surfactant within the airspace. The lung interstitium was apparently enlarged, and the fibroblasts and contractile interstitial cells frequently contained lipid droplets. These alterations were more pronounced and occurred at a faster rate (4 weeks) in diabetes associated with hyperlipidaemia. The structural modifications reported in this study support the functional disturbances observed in association with hyperglycaemia, sustaining the conclusion that the lung is an organ affected by diabetes.     

An entire functional mammary gland may comprise the progeny from a single cell

Any epithelial portion of a normal mouse mammary gland can reproduce an entire functional gland when transplanted into an epithelium-free mammary fat pad. Mouse mammary hyperplasias and tumors are clonal dominant populations and probably represent the progeny of a single transformed cell. Our study provides evidence that single multipotent stem cells positioned throughout the mature fully developed mammary gland have the capacity to produce sufficient differentiated progeny to recapitulate an entire functional gland. Our evidence also demonstrates that these stem cells are self-renewing and are found with undiminished capacities in the newly regenerated gland. We have taken advantage of an experimental model where mouse mammary tumor virus infects mammary epithelial cells and inserts a deoxyribonucleic acid copy(ies) of its genome during replication. The insertions occur randomly within the somatic genome. CzechII mice have no endogenous nucleic acid sequence homology with mouse mammary tumor virus; therefore all viral insertions may be detected by Southern analysis provided a sufficient number of cells contain a specific insertional event. Transplantation of random fragments of infected CzechII mammary gland produced clonal-dominant epithelial populations in epithelium-free mammary fat pads. Serial transplantation of pieces of the clonally derived outgrowths produced second generation glands possessing the same viral insertion sites providing evidence for self-renewal of the original stem cell. Limiting dilution studies with cell cultures derived from third generation clonal outgrowths demonstrated that three multipotent but distinct mammary epithelial progenitors were present in clonally derived mammary epithelial populations. Estimation of the potential number of multipotent epithelial cells that may be evolved from an individual mammary-specific stem cell by self-renewal is in the order of 10(12)-10(13). Therefore, one stem cell might easily account for the renewal of mammary epithelium over several transplant generations.     

Horseradish peroxidase permeation from the capillaries of the stria vascularis after inoculation of endotoxin into the middle ear

Escherichia coli-derived endotoxin was inoculated in the middle ear of guinea pigs 24 hours after being injected intraperitoneally. Twenty-four hours after the middle ear inoculation, horseradish peroxidase (HRP) was injected via the femoral vein and the permeability of HRP through the capillaries of the stria vascularis and the destination of the leaked HRP were examined. A large amount of HRP leaked out of the capillary through he opened endothelial cell junctions and penetrated the enlarged intercellular spaces. Leaked HRP entered the pinocytotic vesicles of the intermediate cells. Even slightly degenerated intermediate cells retained this function. The HRP penetrated the spongelike structure of the marginal cells leading to the intercellular space. This structure was not observed without endotoxin. The HRP could not pass the cochlear duct through the tight junctions between marginal cells. Blood sludging was observed in the strial capillaries. It appeared more frequently in the upper three turns than in the basal turn. The HRP leakage out of the capillaries was observed not only in the upper three turns but also in the basal turn.     

Lipids in tissues of the eye. XII. Phospholipid and sphingomyelin concentrations in bovine lenses in relation to differentiation and ageing

A choroid plexus papilloma (CPP) removed from a child with communicating hydrocephalus was studied with the electron microscope. In addition to the usual organelles, the neoplastic epithelial cells contained polypoid microvilli, cilia, coated pinocytotic vesicles, and apical tight junctions. The basal surfaces were consistently juxtaposed to large capillaries lined by fenestrated endothelium. The fine structural details of the tumor were essentially the same as those of normal CP and thus appeared ideally suited for an active secretory function. Two additional features in this tumor were tubular bodies in the endothelium and well developed pericytes. The present findings provide strong evidence for cerebrospinal fluid production by CPP, a phenomenon that has long been suggested by clinical observation.     

An ultrastructural study of the intimal hyperplasia in healing microarterial anastomoses

The purpose of the study was to investigate the ultrastructure of intimal hyperplastic cells. End-to-end microarterial anastomoses were studied in a rabbit free-tissue-transfer model. There were five experimental groups, with 1, 3, 7, 14, or 28 days follow-up. At sacrifice the anastomoses were tested for patency and then examined by light and electron microscopy. At days 1 and 3 the repaired intima was covered with macrophages and extravasated erythrocytes. At day 7 spindle-shaped fibroblasts with copious rough endoplasmic reticulum were seen. Some of these cells also contained pinocytotic vesicles, filaments with focal densities, and subplasmalemmal attachment sites, the features of smooth muscle cells. At day 14, more cells contained smooth muscle features and these features were also more pronounced. These young myofibroblasts were plumper than adjacent fibroblasts. At day 28 mature myofibroblasts with a full complement of organelles were present. The results, therefore, supported the hypothesis that myofibroblasts are present in the intimal hyperplasia of healing microarterial anastomoses.     

Intracellular behaviour of samarium and europium in lactating mammary gland

The subcellular localization of samarium and europium, two rare-earths, increasingly used in both medical and industrial fields, has been studied in several organs such as liver and kidney but never in the mammary gland despite of its importance in the biology of lactation and nutrition domains. The intracellular behaviour of samarium and europium after their intra-peritoneal administration in the lactating mammary gland cells was investigated. The results showed the presence of very electron dense deposits in the glandular epithelial cell lysosomes. These particular lysosomes were never observed in the mammary cell lysosomes of control rats. These intralysosomal deposits were probably composed of insoluble samarium or europium phosphates by analogy with previous studies, the transmission electron microscopy, the ion mass microscopy and the electron probe microanalysis, and other techniques allowing the identification of the chemical structure of the intralysosomal deposits.     

Paced Auditory Serial Addition Test: adult norms and moderator variables

The structure of the capillary endothelium in the pecten oculi of the domestic fowl was investigated by scanning and transmission electron microscopy. Scanning electron microscopy results demonstrated the existence of a vast array of irregular microplicae that projected from the luminal surface of the capillary endothelium. In between these microplicae were numerous crevices. The microplicae were closely packed and showed no preferred orientation regarding either the longitudinal or transverse plane of the capillaries. Transmission electron microscopy revealed the section profiles of the microplicae: their tortuity, branching, interdigitations and the magnitude of the crevices contained. The endothelial cytoplasm exhibited a few mitochondria and micropinocytotic vesicles. The apparent set-up of the luminal plasmalemmal infoldings seemed to be designed for effecting impedance to the pectineal blood flow and thereby facilitating passive permeation of blood-borne nutrients to the inner neural retina. The reasons of such passive transport operation mediated by the endothelial microplicae of the avian pecten oculi are discussed in the perspective of the existing literature.     

Morphological and biochemical status of the mammary gland as influenced by conjugated linoleic acid: implication for a reduction in mammary cancer risk

Previous research showed that treatment with conjugated linoleic acid (CLA) during the period of active mammary gland morphogenesis was sufficient to confer a lasting protection against subsequent mammary tumorigenesis induced by methylnitrosourea. The present study was designed to characterize certain morphological and biochemical changes of the mammary gland that might potentially render it less susceptible to cancer induction. Female Sprague Dawley rats were fed a 1% CLA diet from weaning until about 50 days of age. The mammary gland parameters under investigation included (a) the deposition of neutral lipid, (b) the identification and quantification of CLA and its metabolites, (c) the density of the epithelium, and (d) the proliferative activity of various structural components. Our results showed that CLA treatment did not affect total fat deposition in the mammary tissue nor the extent of epithelial invasion into the surrounding fat pad but was able to cause a 20% reduction in the density of the ductal-lobular tree as determined by digitized image analysis of the whole mounts. This was accompanied by a suppression of bromodeoxyuridine labeling in the terminal end buds and lobuloalveolar buds. The recovery of desaturation and elongation products of CLA in the mammary gland confirmed our prior suggestion that the metabolism of CLA might be critical to risk modulation. The significance of the above findings was investigated in a mammary carcinogenesis bioassay with the use of the dimethylbenz[a]anthracene model. When CLA was started at weaning and continued for 6 months until the end of the experiment, this schedule of supplementation produced essentially the same magnitude of mammary tumor inhibition in the dimethylbenz[a]anthracene model as that produced by 1 month of CLA feeding from weaning. The observation is consistent with the hypothesis that exposure to CLA during the time of mammary gland maturation may modify the developmental potential of a subset of target cells that are normally susceptible to carcinogen-induced transformation.     

Lung cancer

This report documents a rare variant of mammary hamartoma with presence of lobules of brown adipose tissue. The lesion was discovered in a control mammogram performed in a 42-year-old woman 15 months after hormonal substitute therapy for radical hysterectomy with bilateral salpingo-oophorectomy. Complete excision is the adequate therapy. The presence of brown fat in the hamartoma should be considered in the differential diagnosis of the exceptional hibernoma of the breast and other tumors composed of large cells with granular or multilocular cytoplasm, such as lipoblastoma, sebaceous adenoma, granular cell tumor and histiocytoid or lipid rich carcinomas.     

Activation and function of the epidermal growth factor receptor and erbB-2 during mammary gland morphogenesis

The hormonal stimulation of mammary gland morphogenesis is believed to occur through growth factor receptor signaling pathways. To determine the importance of the epidermal growth factor receptor (EGFR) pathway, we examined extracts of inguinal mammary glands from prepubertal and pubertal mice for tyrosine-phosphorylated EGFR and other erbB receptors. Tyrosine phosphorylation of both EGFR and erbB-2 was detected in normal female BALB/c mice at 5-6 weeks of age, but not during the prepubertal stage, e.g., 24 days of age. Treatment of mice with estradiol or epidermal growth factor also stimulated the formation of mammary EGFR/erbB-2 phosphotyrosine. Waved-2 mice, which have impaired EGFR kinase activity, exhibited less mammary development than did wild-type (wt) mice when both were evaluated at 36 days of age. Because EGFR knockout (KO) mice die shortly after birth, glands from the newborns were implanted under the renal capsules of female nude mice. Under these conditions, extensive ductal growth was observed in mammary glands from wt animals; in contrast, glands from EGFR KO mice failed to grow beyond rudimentary structures. Tissue recombinants revealed that the wt fat pad supported the morphogenesis of EGFR KO epithelium, whereas the EGFR KO fat pad did not. Taken together, these data suggest that EGFR is essential for morphogenesis of the mammary ducts and functions during this period of mammary development as a heterodimer with erbB-2 in the mammary stroma.     

Histo- and cytophysiology of the lactating mammary gland of the African elephant (Loxodonta africana)

The lactating mammary gland of the African elephant (Loxodonta africana) has been studied with a panel of morphological techniques focusing on (1) the functional changes during the secretory process, (2) proliferative process [by application of proliferating cell nuclear antigen (PCNA) immunohistochemistry] and apoptotic phenomena [by use of the TUNEL technique] in the individual lobules, and (3) components of milk and milk-fat-globule membrane. In the lactating gland, the lobules are variably differentiated; within a lobule, however, the alveoli are usually similarly differentiated. The morphology of their alveoli suggests a classification of the lobules into types 1-3. Lobules of type 1 are composed of immature tubular alveoli with mitotic figures and numerous PCNA-positive nuclei; advanced type 1 alveoli contain abundant glycogen and specific secretory granules. Lobules of type 2 are further subdivided. In type 2a lobules, the epithelial cells of the alveoli form tall apical protrusions, which in part are occupied by small lipid droplets and which are pinched off in an apocrine fashion. The number of lysosomes varies considerably. Type 2b is the most common type, with striking basal membrane foldings, abundant rough endoplasmic reticulum cisterns, large Golgi apparatus, numerous mitochondria, lipid droplets, and protein vesicles with 30- to 90-nm-wide casein micelles. The lipid droplets are pinched off with minimal amounts of cytoplasm. Type 2c is composed of alveoli with a cuboidal epithelium and few signs of secretory activity. Increasing expression of peanut-agglutinin-binding sites parallels the maturation and differentiation of the glandular cells. Type 3 lobules are marked by numerous TUNEL-positive nuclei and large lipid droplets and are apparently degenerating structures. Cytokeratin (CK) 14 is usually present in the myoepithelial cells; CK 19 and CK 7 mark ductal and immature alveolar epithelia. Milk protein content varies between 2.6% and 6.3%, and casein micelles range from 35 to 90 nm in diameter. The diameter of intra-alveolar milk fat globules ranges from 5 to 25 micrometer and the membranes bear a filamentous surface coat composed of membrane-anchored mucins; gel-electrophoretic analysis of these mucins from different individuals demonstrates the presence of mucin MUC 1, which is expressed with considerable genetic heterogeneity.     

Calcium-induced intercellular adhesion of keratinocytes does not involve accumulation of beta 1 integrins at cell-cell contacts and does not involve changes in the levels or phosphorylation of catenins

The development of the lymphatic system in the rat diaphragm was studied from embryonic day 16 to 25 weeks after birth by histochemistry for 5'-nucleotidase, scanning electron microscopy of KOH-treated or intact tissues, and transmission electron microscopy of thin sections. On embryonic day 16, distinct lymphatics were noted in the subpleural space of the diaphragm periphery. The endothelial cells at this stage contained an abundance of rough endoplasmic reticulum, a developed Golgi apparatus and mitochondria, and fewer pinocytotic vesicles than those in adults. The subpleural lymphatics subsequently increased and formed a polygonal network. They possessed many valves, and by postnatal week 6, some thick collecting lymphatics became endowed with smooth muscle cells. On embryonic day 19, some lymphatics appeared in the subperitoneal space. They extended centripetally and had many lateral projections that subsequently became elongated and connected with those from adjacent lymphatics, thus forming a lattice-like network. During the early postnatal days, the subperitoneal lymphatics projected many bulges that subsequently became elongated, and came into contact with the pores among the mesothelial cells, thus forming lymphatic stomata connecting the lymphatic lacunae to the peritoneal cavity. The lymphatic stomata increased until postnatal week 10. The results show that lymphatics appear as early as embryonic day 16 in the subpleural space of the diaphragm periphery, and develop with age by sprouting to form networks in both the subpleural and the subperitoneal spaces, and that the direct connection of the lymphatic lacunae to the peritoneal cavity is formed after birth.     

Proliferation, development and DNA adduct levels in the mammary gland of rats given 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine and a high fat diet

2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is a heterocyclic amine derived from cooked meat that is a mammary gland carcinogen in rats. A carcinogenic dose-regimen of PhIP (75 mg/kg, p.o., 10 doses, once per day) was administered to 43-day old female Sprague-Dawley rats, and the rats were then placed on a defined high fat (23.5% corn oil) or low fat (5% corn oil) diet for up to 6 weeks. At various times after carcinogen and diet, and prior to carcinogenesis, we examined the percentage of proliferating cells in terminal end bud (TEB) epithelial structures of the rat mammary gland by proliferating cell nuclear antigen staining, mammary gland architecture by whole mounting, and PhIP-DNA adduct levels in mammary epithelial cells by the 32P-post-labeling assay. Immediately after dosing, the percentage of proliferating epithelial cells in TEBs was significantly higher in PhIP-treated rats than in control rats receiving vehicle only [7.5 +/- 0.9% (n = 99) versus 4.2 +/- 0.6% (n = 127), respectively]. The mammary glands of PhIP-treated rats showed a significantly lower density of alveolar buds (ABs) and a higher density of TEBs than control rats, which suggests that PhIP exposure partially inhibited the normal glandular differentiation of TEBs to ABs. After 6 weeks on the diet, proliferation in TEBs was statistically higher in rats given PhIP plus a high fat diet than in rats given vehicle plus a low fat diet. The mammary glands from rats on a high fat diet also showed a statistically higher density of TEBs when compared with rats on a low fat diet [2.08 +/- 0.34% versus 1.04 +/- 0.20%, respectively (n = 6)]. PhIP-DNA adduct levels were relatively high in mammary epithelial cells of treated rats. At 3 h after the last dose of PhIP, DNA adduct levels [relative adduct labeling (RAL) x 10(7), mean +/- SE] were 10.5 +/- 1.7 (n = 8) and 0.9 +/- 0.2 (n = 7) in epithelial cells isolated from mammary gland and in the liver, respectively. DNA adduct removal rates from the mammary gland were not different between rats on the high fat and low fat diets. Adducts were still detected after 6 weeks on either diet. Thus, events that occurred prior to neoplasia in the mammary glands of PhIP-treated rats include formation of PhIP-DNA adducts at relatively high levels, and enhanced proliferation in TEBs (putative sites of origin of mammary gland carcinomas) and partial inhibition of TEB differentiation. The high fat diet, a promoter of PhIP-induced mammary gland carcinogenesis, appeared to sustain the proliferative effect of PhIP in mammary gland TEBs at a time when PhIP-DNA adducts are still detectable. These early events may contribute to the targeting and carcinogenicity of PhIP to the mammary gland of rats.     

Light microscopic and ultrastructural observations of a metastasizing malignant epithelioid schwannoma

The light microscopy and ultrastructure of a malignant epithelioid schwannoma are described. Characteristic cells resembling perineural elements with various degrees of differentiation were observed. Primitive epithelioid cells contained scant ergastoplasm, and few tubules and filaments, but did have abundant free ribosomes and Golgi membranes. Also noted were junctional complexes and focal fusion of plasma membranes, basal laminae were absent. Better differentiated cells were completely limited by a well-developed basal membrane and had an abundance of intracytoplasmic filaments and multiple pinocytotic vesicles. The intercellular ground substance was composed of numerous fine collagen fibrils and amorphous, basement membrane-like, electron-dense material. A striking ultrastructural similarity of the tumor cells to those encountered in ethylnitrosourea-induced malignant schwannomas in rats was noteworthy.