Abstract: Mammalian spermatogenesis reflects a dynamic partnership between somatic genes which display altered expression patterns within the testes and germ cell specific genes expressed solely during male gametogenesis. Among the latter include the haploid-specific transition proteins and protamines which facilitate the molecular restructuring of the chromatin during the terminal differentiation of the spermatid nucleus. Previously, we have shown that the human protamines PRM1, PRM2 and transition protein TNP2 transcripts persist in mature spermatozoa subsequent to their functional role during spermiogenesis. We now demonstrate the conserved presence of these same spermatozoal mRNAs in mouse. While the basis for the persistence of these haploid-specific transcripts remains poorly defined, the results presented firmly establish that the presence of mRNA in mammalian spermatozoa is a general phenomenon. The potential use of these transcripts as a tool for investigating molecular pathologies is discussed.

Abstract: The objective of the present study was to analyze the prospective alterations of the testis and epididymis in a defined strain of alcoholic rats in order to contribute to our understanding of the effects of chronic alcoholism on reproduction. The testis and epididymis of the animals were submitted to morphological analysis by macroscopy, light microscopy and electron microscopy and to morphometric analysis. The UCh rats showed atrophy of the epithelium and reduction of testis and epididymis weight, liver hypertrophy and fat infiltration and alterations of the hypothalamus-pituitary axis. Ethanol induces changes in the weight and in the epithelium of the testis and epididymis and in the hypothalamus-pituitary axis of the UCh rats.

Abstract: The young spermatid of Sudarikovina taterae exhibits a nucleus with a partially condensed chromatin and a differentiation zone, bordered by cortical microtubules, delimited at the front by arched membranes and containing two centrioles The latter are parallel to one another and linked together at their bases by electron-dense material During spermiogenesis, one of the centrioles gives rise to a flagellum whereas the other disappears Crested-like bodies lie outside the cortical microtubules and the nucleus migrates in the spermatid along the axoneme At the end of spermiogenesis, the ring of arched membranes constricts and the old spermatid becomes detached from the residual cytoplasm The S taterae mature spermatozoon is filiform and tapered at both extremities It exhibits an apical cone of electron-dense material and seven crested-like bodies 50 to 100 nm thick The cortical microtubules run along the whole length of the spermatozoon They are spiralized at their anterior extremities and straight over the rest of their length The nucleus is a fine compact cord interposed between the axoneme and the cortical microtubules The cytoplasm is slightly electron-dense in regions I and II of the gamete In regions III, IV and V, it contains fine electron-dense granulations and patches of electron-lucent material An electron-dense material located both between and under centrioles has not been observed before in a platyhelminth Similarly, distinctive ultrastructural characters between centrioles and seven crested-like bodies have not been previously described in a cestode Moreover, we report for the first time the existence of cortical microtubules spiralized only at their anterior extremities in a cestode from a mammal

Abstract: The spermiogenesis and the spermatozoon ultrastructure of Sorubim lima were studied. Our observations showed that early spermatids are round-shaped cells, have spherical nucleus with diffuse chromatin, small quantity of mitochondria and large amount of vesicles in the cytoplasm. During the differentiation process in the nucleus, chromatin compacts in a progressive and homogeneous way, and the flagellum is formed. In the cytoplasm the vesicles, that have double membranes, aggregate and fuse on the plasma membrane. The spermatozoa of S. lima have no acrosome and show spherical nucleus with homogeneous and highly compacted chromatin, intermediary piece with mitochondria and double wall vesicles contiguous to the plasma membrane, as well as a flagellum formed by a basic axoneme (9 + 2).

Abstract: Human cytomegalovirus (HCMV) is capable of infecting human bone marrow fibroblastic stromal cells (HBMF-sc). This infection is important to assess in regard to the pathogenesis of HCMV, particularly in immunocompromised patients. Stromal fibroblastic cells were infected by Towne strain of cytomegalovirus (CMV) in vitro. Several ultrastructural features of uninfected HBMF-sc were also described. The CMV-infected cells exhibited significant mitochondrial enlargement, production of dense bodies by the Golgi apparatus and cytoplasmic accumulation. Ultrastructural aspects of HCMV entry in host cells, capture by endosomes, penetration of genetic material in the nucleoplasm, assembly and formation of nucleocapsids were detected and described. Viral fusion and transit through the nuclear envelope were shown along with envelope proliferations. Trafficking of virions, maturation and completion of their cytoplasmic coating were also illustrated. Fully developed virions, defective virions, other apparently-emptied vesicles, multivesicular bodies as well as cytoplasmic dense bodies were illustrated along arrays of microtubules organized by defective centrosomes and constituted a peculiar structure that we termed 'viral field'. While some viral and dense bodies were carried to adjacent sites of the plasmalemma, in order to be extruded from the infected cells, others were concentrated into black holes--dense heterogenous bodies accumulated at the periphery of viral fields. This study further described the functional aspects of HBMF-sc and summarized the unknown aspects of ultrastructural characteristics of HCMV-infected fibroblastic stromal cells which may serve as harmful reservoir for the replication of virus. In addition, the findings of this study may stimulate further investigations about the basic cell biology and functions of the bone marrow stromal cells and may also generate some interests to bone marrow transplantation medicine as to how HBMF-sc can serve as a reservoir in the pathogenesis of CMV infections.

Abstract: Regenerating tail fins were studied in two species of teleosts, Tilapia rendalli and Cyprinus carpio, treated with indomethacin, aspirin, dexamethasone, penicillamine, and beta-aminoproprionitrile, drugs known to disrupt collagen metabolism in mammals. Collagen was studied under the light microscope by the Picrosirius-polarization method and also under the electron microscope. In general, these drugs disturbed the deposition and organization of collagen fibrils leading to abnormally thin or practically absent lepidotrichia and actinotrichia, and also to disorganized fibrous connective tissue. The resulting disorganization of the collagenous scaffolding of the regenerating dermoskeleton was probably responsible for a secondary effect on blastema distalization and on the general fin ray patterning that were also observed. The foregoing observations suggest that the stromal histoarchitecture of the regenerate plays a vital role in fin regeneration and indicate that these drugs may be useful in studying the extracellular matrix-cell interactions at the cellular and molecular level. In addition, the present findings provide a basis for developing different biological models by using teleost fin regeneration.

Abstract: Neoplastic progression is a prolonged and stepwise process, while tumor growth occurs after a series of molecular alterations that culminate in tumorigenesis. The phenotypic changes of transformation in breast carcinogenesis were studied through the use of scanning and transmission electron microscopy. MCF 10F, a spontaneously immortalized human breast epithelial cell line (Soule et al., 1990), was treated with 7,12 dimethylbenz(a)anthracene (DMBA) (Calaf and Russo, 1993) and then transfected with the c-Ha-ras oncogene (Calaf et al., 1995). Treated cells showed a progression in altered morphology, anchorage independency, invasiveness and tumorigenicity in the SCID. Scanning and transmission electron microscopy illustrated that the transformed cells could be distinguished from control cells by the expression of morphological characteristics such as loss of contact inhibition, irregular size and shape, emission of long filopodia and formation of stratified layers. In contrast, control cells showed uniform, flattened and polyhedrical cells, well closely juxtaposed to each other and joined by cytoplasmic interdigitations. Control cells also did not form colonies in agar-methocel, and were not invasive or tumorigenic in SCID mice. These studies showed the progression of breast carcinogenesis by phenotypical changes induced by the carcinogen and the insertion of the c-Ha-ras oncogene.

Abstract: The present paper reports modifications in the electrophoretic and cytochemical characteristics of mature and immature stallion spermatozoa. Some sperm surface glycoproteins (36, 32, 29, 21, 20, 18 kDa) detected in cauda epididymidis spermatozoa, were either absent or present in a very low relative concentration in immature sperm cells. A major 14 kDa protein band, observed in sperm extracts obtained from ductus efferentes, progressively decreased along the epididymal ductus. The nature and distribution of carbohydrate residues on the sperm membrane, during epididymal maturation, was also studied by use of lectin probes. Some protein bands bound concanavalin A while others, as the 36, 32 and 20 kDa proteins, exhibited higher affinity for WGA lectin. The distribution and relative density of mannose-, galactose-, N-acetylglucosamine-, N-acetylgalactosamine-, fucose- and sialic acid-containing macromolecules showed a characteristic pattern depending on the sperm membrane domain and on its origin. Some sperm surface domains displayed affinity for more than one lectin, indicating a diversity in their exposed carbohydrate residues, whereas others bound only one or no lectin. The passage of spermatozoa through the epididymidis was accompanied by changes in the accessibility or abundance of lectin ligands. Some lectins (UEA, WGA, LPA) gave stronger reaction in mature spermatozoa, while others (RCA, WFH, PNA) stained better immature spermatozoa. This remodeling of sperm surface molecules is probably a consequence of interactions between spermatozoa and the epididymal secretions, and may reflect addition or adsorption of new molecules, space configurations changes or biochemical modifications of pre-existing compounds. Our results suggest that the distribution and density of terminal oligosaccharidic residues on the sperm plasma membrane have species-specific characteristics. These post testicular developmental changes may be of significance in the overall understanding of the stallion fertility.

Abstract: The spermatozoa of Uta stansburiana and Urosaurus ornatus show the following squamate autapomorphies: a single perforatorium extending anteriorly from the apical tip of the paracrystalline subacrosomal cone; the presence of an epinuclear electron lucent region; intermitochondrial dense bodies; and the fibrous sheath extending into the midpiece. The acrosome vesicle is flattened and concentrically zoned apically; basally it overlies a subacrosomal cone which invests the nuclear rostrum. A stopper-like perforatorial base plate, rounded nuclear shoulders and a basal nuclear fossa are present. The proximal centriole contains a density within its centre for approximately one half its length and lies at approximately 80 degrees to the distal centriole. The two central singlets of the axoneme extend into the short distal centriole. A peripheral dense fibre is associated with each of the nine triplets of the distal centriole, and the fibre continues posteriorly with each of the nine doublets of the axoneme. A central fibre is associated with the two central singlets. All fibres are absent or vestigial at the level of the annulus. Mitochondria are short sinuous with a maximum of eight seen in transverse section. Uta and Urosaurus sperm differ from each other in their arrangement of intermitochondrial dense bodies in two ways: 1) longitudinally, Uta has five incomplete 'rings' of dense bodies, whereas Urosaurus has only four such rings; 2) in cross section, each individual 'ring' of Uta may contain up to four irregularly spaced dense bodies, whereas Urosaurus contains a maximum of only two dense bodies. The sperm of Uta and Urosaurus show strong similarities to those of the agamids and polychrotids. No spermatozoal autapomorphies for the Phrynosomatidae were found.

Abstract: Granulation tissue involved in tissue repair and in the stroma reaction to epithelial tumors is characterized by the presence of myofibroblastic cells. It has been previously reported that granulocyte macrophage-colony stimulating factor (GM-CSF) induces a fibrotic reaction containing numerous myofibroblasts. This reaction results from a cascade of events, including stimulation of transforming growth factor-beta1 (TGF-beta1) production by macrophages which, in turn, promotes alpha-smooth muscle actin and collagen synthesis by fibroblasts. Moreover, GM-CSF is known to be expressed by many tumor cell types. In this study we have analyzed, by means of reverse transcription-polymerase chain reaction, GM-CSF mRNA expression in a progressive and a regressive rat colon carcinomas and in the corresponding cell lines, eliciting different degrees of desmoplastic reaction. We have also evaluated the expression of GM-CSF protein in selected cases. The expression of GM-CSF mRNA and, when tested, protein were higher in progressive compared to regressive cancer cells both in vivo and in vitro. We then investigated GM-CSF mRNA and protein expression in different human colon cancer cell lines known to exhibit different degrees of aggressivity in vivo. We found high levels of GM-CSF mRNA and protein in the most aggressive cell lines. Similar results were also obtained on human breast and cervical cancer cell lines. Our results are in agreement with the assumption that GM-CSF expression is correlated to tumor aggressivity. Conceivably, one of the GM-CSF actions affecting tumor progression is exerted through its influence on stroma reaction development.

Abstract: The neoplastic conversion of a normal cell to a malignant one is a multistage process that occurs after a series of molecular alterations Several chemical and physical agents can alter the morphology of different types of cells Scanning and transmission electron microscopy have been valuable in evaluating changes that occur in the progression of transformation MCF-10F, a spontaneously immortalized human breast epithelial cell line (Soule et al, 1990), was treated with benz(alpha)pyrene (BP) (Calaf and Russo, 1993) and then transfected with the c-Ha-ras oncogene (Calaf et al, 1995) The phenotypic changes of breast cancer progression were studied through the use of scanning and transmission electron microscopy Activated oncogenes have been detected in a variety of malignant tumors and the altered expression of certain genes seems to play a role in the cancer process Carcinogen-treated and transfected cells showed a progression of changes in the morphology, anchorage independent growth, invasiveness and capability of tumor formation in the SCID mice This in vitro cancer model can parallel the progression of breast cancer seen through molecular changes that occur and have been observed during the natural development of this disease

Abstract: The spermatozoon of Solea senegalensis (Kaup, 1858) consists of an acrosome-less ovoid head, a short midpiece containing several irregular mitochondria embedded in the cytoplamic mass, and a long tail with two lateral fins and a conventional 9 + 2 axoneme. The centrioles are housed in a deep nuclear fossa and are both orientated in the same longitudinal axis of the spermatozoon. The overall structure of this spermatozoon conforms to the sperm type considered to be plesiomorphic in the neopterigians (type I sperm). The likely apomorphic (coaxial) orientation of the centrioles defines the spermatozoal morphology of the Soleidae investigated thus far and separates them from the other known pleuronectiform spermatozoa.

Abstract: The type of synaptic terminals from the cochlear nucleus and inferior colliculus that terminate in the contralateral ventral cochlear nucleus are not known. These terminals were studied with the electron microscope and immunogold after injection of wheat germ agglutinin conjugated to horseradish peroxidase into the inferior colliculus or into the cochlear nucleus. The tracer anterogradely labelled boutons onto the main neurons of the contralateral ventral cochlear nucleus. Most of these cells (95%) were glycine immuno-negative and represent excitatory neurons. After injection of the tracer into the contralateral inferior colliculus few anterogradely labelled boutons were seen on spherical and multipolar cells of type II in the anteroventral cochlear nucleus. Rare labelled boutons were present on multipolar cells of type I and II, globular neurons and octopus cells in the posteroventral cochlear nucleus. After injection into the contralateral dorsal and ventral cochlear nucleus labelled boutons were seen more frequently than after injection into the inferior colliculus. These terminals contacted most of large neurons, especially multipolar cells of type II and less frequently of type I. Also globular and spherical cells were contacted by commissural terminals. Octopus cells received less frequently putative commissural terminals. Most boutons contained pleomorphic vesicles and stored GABA. A lower number of boutons with pleomorphic and flat vesicles contained glycine and sometimes GABA, both inhibitory neurotransmitters. Few boutons containing round vesicles were immuno-negative for both glycine and GABA, and were considered putative commissural excitatory terminals. The latter often contacted glycinergic neurons of type II so that also these terminals might elicit an inhibition with at least a disynaptic mechanism after contralateral stimulation.

Abstract: The purpose of this study was to assess the expression of cell adhesion molecules ICAM-1 (intercellular adhesion molecule-1) and VCAM-1 (vascular cell adhesion molecule-1) in endothelial cell-derived foam cells. Hamster aortic endothelial cells (HAEC) in culture were exposed to hypercholesterolemic or normal homologous serum for 24 h. At the end of the incubation period, HAEC exposed to hypercholesterolemic serum exhibited numerous lipid droplets and had a general aspect of foam cells. When examined for the expression of ICAM-1 and VCAM-1 (by indirect immunofluorescence) normal HAEC expressed constitutively (to low level) on their surface these adhesion molecules; however HAEC-derived foam cells failed to display any labeling. To further assess these results, HAEC were first incubated with normal or hypercholesterolemic sera (as above) and then exposed to freshly isolated normal hamster blood monocytes. These experiments showed that monocytes adhered in small number to normal cells and failed to adhere to the surface of HAEC-derived foam cells. Together these data indicate that endothelial cell-derived foam cells: a) do not express ICAM-1 and VCAM-1 on their surface; b) have low or no adhesion properties for monocytes and c) may represent an appropriate experimental model to study the cellular alterations that take place in the advanced stages of atherosclerosis.

Abstract: There are large numbers of macrophages in the uterus of normal mice. During estrus and metestrus endometrial epithelial cells express macrophage colony-stimulating factor (M-CSF). Numbers of uterine macrophages showed cyclic changes and accumulated beneath the endometrial epithelial cells at estrus and metestrus. However, numbers of uterine macrophages in osteopetrotic (op/op) mice were reduced tenfold compared with normal littermate mice and op/op mouse macrophages were ultrastructurally immature throughout the estrous cycle. Several macrophage chemokines were produced in the uterus. However, administration of antibody against the macrophage colony stimulating factor receptor (c-fms) severely diminished the number of uterine macrophages in normal littermate mice, implying that M-CSF plays a major role in macrophage recruitment in the uterus. Endometrial epithelial cells underwent apoptosis at estrus and metestrus and were disposed into the uterine lumen in op/op mice and littermate mice. In littermate mice a large number of macrophages accumulated beneath the endometrial epithelial cells and actively removed apoptotic cells. In contrast, there were few macrophages in op/op mice and clearance of apoptotic cells by macrophages was defective in the uterus of these mice. Thus M-CSF plays important roles in the recruitment and differentiation of macrophages and in scavenging effete epithelial cells in the uterus.

Abstract: The experimental model of Golden Syrian hamster subjected to concomitant hyperlipemia (diet-induced) and diabetes (by streptozotocin injection) for 24 weeks is characterised by the prevalence of micro- and macroangiopathies. We have used the hyperlipemic-diabetic (HD) hamsters to investigate: a) whether there is an alteration in the reactivity of the resistance arteries (mean internal diameter: 210-250 microm), b) if present, which are the structural and biochemical changes that accompany the functional modifications, and c) to examine the pathomorphological changes induced by the association of hyperlipemia and diabetes on vital organs such as myocardium and kidney glomeruli. To these aims, biochemical assays of plasma components, light- and electronmicroscopy, myographic, morphometric and spectrofluorimetric techniques were used. The mesenteric resistance arteries of HD hamsters exhibited (as compared to similar arteries in normals) a decreased contractile response to noradrenaline (1.86+/-0.35 vs. 2.43+/-0.21), and an impeded endothelium dependent relaxation to acetylcholine (approximately 61.40% vs. approximately 79.80%). The association of hyperlipemia with diabetes induced changes in morphology of the resistance arteries consisting in approximately 10% increase of the intima plus media cross-sectional area, approximately 20% decrease of the vascular lumen area, and approximately 2.85 fold augmentation of the wall to lumen ratio. The resistance arteries exhibited structural modifications of the endothelium (up to 8 copies of Weibel-Palade bodies/endothelial cell), and smooth muscle cells (secretory phenotype), and in the vessels media small calcification cores appeared embedded in a hyperplasic extracellular matrix. The vascular mesenteric bed of the HD hamsters contained approximately 2.30 and approximately 1.30 fold increased concentrations of AGE-collagen and pentosidine, respectively, above the normal values. The HD hamsters displayed also modifications that may be dependent on or may lead to an increase in blood pressure, such as: a) approximately 2 fold increase in the activity of serum angiotensin converting enzyme; b) approximately 4.8 fold enhancement of erythrocytes fragility (as a measure of the oxidative stress); c) left ventricular hypertrophy associated with a progressive disarray of cardiomyocyte contractile fibers, interruptions of the Z bands, and accumulation of collagen-rich extracellular matrix indicative of interstitial fibrosis; d) the kidney glomerular capillaries appeared partially or totally collapsed, with a thickened basement membrane which appeared polymorphic, and in some locations made up of successive layers connected by fine bridges and intercalated nodules; in addition, an increase (approximately 1.50 fold) of the mesangial volume was indicative of glomerulosclerosis.

Abstract: The microanatomy and ultrastructure of the excretory system of Pneumoderma sp. (Gymnosomata) and Creseis virgula Rang, 1828 (Thecosomata) have been investigated by means of semithin serial sections, reconstructions and transmission electron microscopy. The studies revealed a functional metanephridial system consisting of a heart with a single ventricle and auricle in a pericardial cavity and a single kidney in both species. Podocytes in the atrial wall of the pericardial epithelium are the site of ultrafiltration, whereas the flat epithelium of the kidney with numerous basal infoldings and a dense microvillous border on the luminal surface suggests modification of the ultrafiltrate. In Pneumoderma sp., additional loci of ultrafiltration with identical fine structure (meandering slits with diaphragms covered by extracellular matrix) occur in the solitary rhogocytes (pore cells). The presence of podocytes situated on the atrial wall in representatives of two higher opisthobranch taxa contradicts former ideas on the loss of the primary site of ultrafiltration in the ancestors of the Opisthobranchia.

Abstract: In previous studies we demonstrated that loud noise exposure induces ultrastructural alterations in the rat myocardium together with an increase in noradrenergic activity and in mitochondrial calcium influx. To verify the causal relationship between myocardial calcium entry and ultrastructural alterations induced by loud noise, in the present study we coupled routine electron microscopy with cytochemistry specifically dedicated to visualize calcium accumulation (revealed as antimonate deposits). We observed that the ultrastructural alterations occurring in both atrium and ventricle after 12 h of noise exposure, were densely packed with antimonate deposits. In particular, enlargements of the sarcoplasmic reticulum and dilution of the mitochondrial matrix, observed during routine electron microscopy, were markedly positive for calcium accumulation when observed by using antimonate. The present data strongly suggest that calcium entry results in accumulation of this ion at myocardial subcellular level. Moreover, the present results joined with previous evidence indicate that calcium accumulation is the final common pathway responsible for noise-induced myocardial morphological alterations.

Abstract: The ultrastructural examination of liver biopsies from five male cocaine users showed hepatocytes presenting diverse alterations in rough and smooth endoplasmic reticulum, mitochondria, nuclei and microvilli Lipid deposition and an increase of autophagic vacuoles were also observed This study demonstrates that the hepatocyte is an important target cell for cocaine toxic effects in some patients

Abstract: The microanatomy and ultrastructure of the larval excretory system of Patella vulgata L., 1758 has been examined by means of semithin and ultrathin serial sections, reconstructions, and transmission electron microscopy. The protonephridial system appears after torsion and consists of two terminal flame bulbs with narrow, ciliated ducts. Whereas the polyciliary terminal cells (cyrtocytes) are only slightly asymmetrically placed below the mantle cavity, the distal excretory ducts and their openings show remarkable asymmetry due to torsion. Further larval ultrafiltration sites with identical fine-structure (meandering slits with diaphragms covered by extracellular matrix) are present in the solitary rhogocytes (pore cells). The presence of larval protonephridia is regarded as plesiomorphic for Mollusca and the Trochozoa (Spiralia) as a whole and the specific conditions (asymmetry, simplicity) in Patella are probably plesiomorphic for the Gastropoda.

Abstract: Keratinocyte proliferation in normal human skin was induced by application of irritants: sodium lauryl sulfate (SLS) or non-anoic acid (NAA). Each irritant was applied at three different concentrations to normal human skin in vivo under occlusion for 24 h. The irritative response was visually assessed for erythema, and the number of cycling cells was calculated from the number of Ki67 positive cells in cryosections from skin biopsies taken at 0, 24, 48, 72 and 96 h. In addition, to determine the elemental content of the epidermal cells during the response, two epidermal strata were examined by X-ray microanalysis (XRMA). Both the application of SLS and that of NAA resulted in erythema, and the number of Ki67 positive cells was increased at 48 h. XRMA revealed an initial increase in the sodium/potassium (Na/K) ratio, indicating cell membrane injury. Due to an increasing potassium (K) concentration the Na/K ratio decreased after 24 h, which is compatible with proliferation, in accordance with the Ki67 data. This was more evident in SLS-stimulated skin, where also the concentrations of magnesium (Mg) and phosphorus (P) increased, an observation commonly made in proliferating cells. It appears that application of SLS results in a period of cell damage lasting about 24-48 h, followed by a period of proliferation. The effects of NAA appear more complicated, and proliferation may take place in parallel with cell damage.

Abstract: Aortocoronary saphenous vein bypass grafts undergo structural alterations within the arterialized vein, resulting in graft stenosis and failure. Areas of the acellular intima contribute to fissuring, cracking and ulceration, while areas of the media become highly vascular but thinned. This study aimed to examine the ultrastructural features of cell death, including apoptosis and necrosis, in non-atherosclerotic areas of the stenotic aortocoronary saphenous vein bypass grafts. Thirteen stenotic vein grafts were obtained at redo coronary artery bypass grafting. The ultrastructural features of cell death were analysed by electron microscopy. Typical features of necrosis, including focal areas of cytoplasmic oedema, plasmalemmal destruction and nuclear condensation with cytoplasmic organelle destruction, were observed throughout the intima and media. Features of apoptosis, including the presence of apoptotic bodies, were also identified in the hyperplastic intima and its adjacent media. Our observations suggest that both apoptosis and necrosis occur in non-atherosclerotic areas of stenotic aortocoronary saphenous vein bypass grafts.

Abstract: The phagocytic process in cells depends on lysosomal enzymes, high-energy metabolism and cellular recognition. In this paper, we investigated the presence of energy and recognition factors in thrombocytes of turtle Phrynopys hilarii (a freshwater South American species). Turtle thrombocytes (P. hilarii) present glycogen - possibly beta particles - dispersed in their cytoplasm and glycoproteins in the cell surface, as well as a large number of enzymes involved in the endocytic process (Pellizzon, 1996). The activity of these enzymes depends on high-energy metabolism and on cellular recognition provided by specific glycoconjugates (Alberts et al., 1994). This metabolic characterization is demonstrated by the large amount of glycogen particles observed in the cytoplasm by Thiery's method. Glycogen labeling was also observed when concanavalin A-peroxidase was used as a marker for thrombocytes and for endocyted charcoal particles. Our results show that these cells have phagocytic ability, suggesting that their function in blood circulation is not limited to aggregation but may also involve a great potential for phagocytosis.

Abstract: A correlative microscopic study of vertebrate cerebellar mossy fiber glomeruli has been carried out to obtain a three-dimensional view of the multisynaptic contacts formed by afferent mossy fibers with the granule and Golgi cell dendrites and by the monosynaptic relationship of Golgi cell axonal ramifications with granule cell dendrites. Samples of mice, hamsters, teleost fishes and human species were studied by means of one of the following procedures: confocal laser scanning microscopy (CLSM), ethanol-cryofracturing technique and conventional scanning electron microscopy (CSEM) and transmission electron microscopy (TEM) by ultrathin sections and freeze-etching replicas. CLSM, by means of montages of z-series of the cerebellar granular layer, provided a new approach to explore mossy fiber trajectory and branching bifurcation pattern and the quantitative relationship between mossy fibers and granule cell dendrites. CSEM and freeze-fracture method for SEM offered a more detailed in-depth, higher resolution image of outer and inner surface organization of mossy fiber glomeruli. TEM, either by ultrathin sections or freeze-etching replicas, was used as complementary technique for proper orientation, comparative purposes and rational identification of pre- and postsynaptic structures. Freeze-etching replicas showed in addition the real extent of glial cell cytoplasm encapsulating the synaptic glomeruli. The integrated microscopy approach offers a new and more comprehensive view of three-dimensional morphology, organization and quantitative aspects of mossy fiber glomeruli.

Abstract: To develop a gerbil model of Helicobacter pylori-induced chronic active gastritis comparable in severity to human lesions, we made acetic acid-induced ulcer in the anterior antral wall and concurrently challenged 1 x 10(8) colony-forming units bacteria per os. At 30 and 60 days after inoculation, the number of viable bacteria colonizing on the surface epithelium of the gastric mucosa was larger in ulcer-bearing animals compared to non-bearing ones. Furthermore, in the former animals, neutrophil and mononuclear cell infiltration as well as lymphoid follicle formation in the lamina propria was more prominent. Electron microscopically, lymphoid follicle-associated epithelium displayed specialized structures. Namely, brush cells interposed between mucous epithelial cells and characterized by prominent microfilament bundles and many apical vesicles or caveola specifically embraced the cluster of intraepithelially invading lymphocytes and macrophage-like cells by the attenuated cytoplasm in an analogous manner to M cells in Peyer's patches. The present study has demonstrated that ulcer formation enhances both H. pylori colonization and lamina propria lymphoid follicle formation and suggested that follicle-associated epithelium might play roles in the delivery of intraluminal antigen.

Abstract: The structural organization of the Michelia figo mature pollen was investigated. The pollen wall consisted of an outer exine and an inner intine, the former being coated by a thin polysaccharide pellicle. The intine comprised three structurally distinct layers that were equally thick throughout the pollen surface. The generative cell (GC) was closely associated with the vegetative cell (VC) nucleus and its periplasm was found to maintain communication with the sporoderm through a complex plasmalemmic cord. In the freeze-fixed pollen a fluffy coat was detected on the cytoplasmic face of the VC plasmalemma bordering the GC. The plastids were present in only the VC and usually contained abundant small starch grains. In a few pollen grains, however, little or no starch existed and in this case one or more electron dense inclusions appeared in the plastids. Microbodies were found in both the VC and GC. In the VC they presumably have a glyoxysomal function as indicated by the numerous lipid droplets in the cytoplasm and the spatial relationship of microbodies with lipid droplets and/or mitochondria. In the GC the function of the microbodies is unclear once this cell had no abundant lipid reserves and the microbodies did not show any preferential relationship with other organelles. The most conspicuous feature of the VC cytoplasm was the high amount of storage vacuoles which displayed a striking different appearance after one and the other of the fixation techniques used. In contrast to the chemically fixed pollen they were quite polymorphic in the freeze-fixed pollen, and appeared uniformly filled with fibrillar material. Enzymatic digestion with protease has revealed most of this material to be proteinaceous in nature. The existence of phytin reserves is, however, also probable. These protein storage vacuoles closely resemble those in storage tissues of seeds and fruits.

Abstract: PCB 128 (2,2',3,3',4,4'-hexachlorobiphenyl) prepared in 4% corn oil and mixed in diets was given to weanling Sprague-Dawley rats. The animals were placed in eight groups, each comprising 10 males or 10 females; each group received a diet that contained 0.05, 0.5 or 5 ppm PCB. Ten animals of each gender that served as the controls were given diets mixed with only corn oil. Thirteen weeks after the commencement of dosing, animals were euthanized and liver specimens were harvested and prepared for transmission electron microscopy. The architecture of the liver parenchymal cell was indistinguishable in the animals of the lowest concentration group from those in the controls. However, smooth endoplasmic reticulum profiles increased, and abnormal mitochondria were noted in the liver of rats, regardless of gender, from 0.5 and 5 ppm groups. Based on our previous work, PCB 128 is estimated to be equally toxic as PCB 153, another di-ortho substituted PCB congener.

Abstract: Peri-vascular matrices having a finely textured granular substructure have been identified in 27 human lesions: these were mostly malignancies but included benign tumours and reactive processes The matrices were defined as stromal components surrounding endothelium and pericytes, and lying between vessels and adjacent lesional cells They were identified as having a finely textured, uniform and moderately dense substructure, and differed from a conventional basal lamina expected at these sites by the absence of the typical lamina densa/lamina lucida configuration By light microscope immunohistochemistry, vessels stained positively for laminin and collagen IV, two of the main proteins characterising a conventional basal lamina The present observations emphasise the following 1) The proteins laminin and collagen IV can be found in peri-vascular locations which have a finely textured granular substructure, and which have clearly defined ultrastructural differences from a conventional basal lamina 2) While conventional light microscope immunohistochemistry demonstrates the presence and cellular location of proteins, electron microscopy is helpful for giving information on their physical organisation 3) Peri-vascular granular matrices have a widespread distribution in malignant tumours but also exist in benign tumours and reactive lesions This paper briefly discusses the possible functions of these matrices as modulators of cell biological processes