PKCε is a negative regulator of PVAT-derived vessel formation.

RATIONALE: Vessel formation is a crucial event in tissue repair after injury. Thus, one assumption of innovative therapeutic approaches is the understanding of its molecular mechanisms. Notwithstanding our knowledge of the role of Protein Kinase C epsilon (PKCε) in cardio-protection and vascular restenosis, its role in vessel progenitor differentiation remains elusive. OBJECTIVE: Given the availability of PKCε pharmacological modulators already tested in clinical trials, the specific aim of this study is to unravel the role of PKCε in vessel progenitor differentiation, with implications in vascular pathology and vasculogenesis. METHODS AND RESULTS: Mouse Peri-Vascular Adipose Tissue (PVAT) was used as source of mesenchymal vessel progenitors. VEGF-induced differentiation of PVAT cells down-regulates both PKCε and p-PAK1 protein expression levels. PKCε overexpression and activation: i) reduced the expression levels of SMA and PECAM in endothelial differentiation of PVAT cells; ii) completely abrogated tubules formation in collagen gel assays; iii) increased the expression of p-PAK1. CONCLUSION: PKCε negatively interferes with vessel progenitor differentiation via interaction with PAK-1.

Motor responses of rat hypertrophic intestine following chronic obstruction.

The present work aims at investigating the changes in motor responsiveness of rat intestine hypertrophied by chronic mechanical obstruction. Motor responses to pharmacological agents and electrical field stimulation (EFS) were studied in hypertrophic ileal segments excised from rats subjected to experimental stenosis (n = 20) and compared with responses of control tissues from sham-operated animals (n = 20). Spontaneous motility and contractile responses to exogenous agents (KCl, acetylcholine and substance P) and EFS (10-s trains every minute, 120 mA, 0.5 ms, 1-10 Hz) were increased in hypertrophic longitudinal segments; however, normalization of motor responses to tissue wet weight revealed a remarkable reduction of contractile efficiency in hypertrophied tissues coupled with a loss of sensitivity to nitric oxide-mediated relaxation. Furthermore, EFS under non-adrenergic non-cholinergic (NANC) conditions unveiled a major role of the cholinergic component over the peptidergic one in the neurogenic contraction of hypertrophic intestine. On the whole, hypertrophic intestinal growth emerges as a dynamic process entailing adaptation of smooth muscle and neuronal structures to the increased functional load imposed by lumen obstruction.

Ultrastructural and three dimensional aspects of the lymphatic vessels of the absorbing peripheral lymphatic apparatus in Peyer's patches of the rabbit.

We studied the absorbing lymphatic peripheral vessels of the Peyer's patches of the small and large intestine of the rabbit by means of light microscopy after injection of Neoprene latex and transmission electron microscopy in order to highlight their topographical distributions to blood vessels as well as the morphologic mechanism of transendothelial passage of the lymphocytes to the lymph. The distribution of absorbing lymphatic vessels originates from the lacteal vessels and the subepithelial mucosal lymphatic network, which continue without interruptions and dilations into the vessels of the interfollicular area which are woven into basket-like networks entwining the medio-basal portion of each lymphoid follicle. The interfollicular area vessels then drain into the large vessels of the tunica submucosa, which in turn drain into the valved precollector vessels of the subserosa by way of intramuscular vessels. TEM revealed the absorbing lymphatic vessels to have a continuous endothelial wall without open junctions, fenestrations, and continuous basal lamina. We observed many lymphocytes wedged in the lymphatic endothelial wall. This underlines the different phases of their migration from the lymphoid tissue in the lumen of the lymphatic vessel. Results of ultrathin serial sections and three dimensional reconstruction of lymphatic vessel segments with included lymphocyte showed the transendothelial passage of lymphocyte, through the "intraendothelial channels."

Histological effect of (R)-alpha-methylhistamine on ethanol damage in rat gastric mucosa: influence on mucus production.

(R)-alpha-Methylhistamine, a selective agonist of histamine H3 receptors, prevents macroscopically visible gastric lesions by absolute ethanol in the rat. A further insight into its activity was the aim of our study. Rats were given saline or (R)-alpha-methylhistamine (100 mg/kg) intragastrically. After 30 min, absolute ethanol was given and gastric mucosa was sampled 60 min later. Histologic damage and intracellular and adherent mucus were quantified. Luminal surface and mucous cells were examined by scanning and transmission electron microscopy. (R)-alpha-Methylhistamine reduced the extent of lesions by ethanol from 96 to 18%. Surface mucous cells and mucous neck cells were increased in volume and number, packaging of intracellular mucus was modified, and the secretory processes were promoted by (R)-alpha-methylhistamine itself, although these modifications were mostly evident in stomachs subsequently exposed to ethanol. Adherent mucus layer thickness was increased by (R)-alpha-methylhistamine only after ethanol exposure. It is concluded that (R)-alpha-methylhistamine predisposes mucous cells to react to ethanol.

[The absorbing peripheral lymphatic apparatus and blood vessels of the vermiform appendix: the 3-dimensional and ultrastructural aspects]. / Appareil lymphatique périphérique absorbant (ALPA) et vaisseaux sanguins de l'appendice vermiforme: aspects tridimensionnels et ultrastructuraux.

We studied the lymphatic angiotectonics of the vermiform appendix of the rabbit, using direct injection of Neoprene latex, electron microscopy and three-dimensional models. We observed that the vermiform appendix is made of absorbing lymphatic vessels distributed among the tubular glands of the tunica mucosa. Moreover, these absorbing lymphatic vessels continue through the straight vessels into a vast, basket-like lymphatic network, which envelopes the medium and basal portions of each single lymphoid follicle. In addition, the vessels of this network reach the submucosal network, whose vessels are peculiar in shape and size. The submucosal network, in turn, drains the lymph through the small caliber intramuscular vessels in the valved subserous precollector lymphatic vessels. We also highlighted the ultrastructural characteristics of the absorbing lymphatic vessels, which completely lack openings at the contacts between the adjacent endothelial cells, which were always joined by specialized junction complexes. Moreover, we reported the presence of intraendothelial channels which we affirm are dynamic morphologic entities, through which lymphocyte migration from the peri-interfollicular lymphoid tissue inside the absorbing lymphatic vessels takes place.

[The fine structure of the gustatory cells of the circumvallate, foliate and fungiform papillae in hibernating bats]. / Sulla fine struttura delle cellule gustative delle papille circumvallate, foliate e fungiformi di pipistrelli ibernanti.

The cells of vallate, foliate and fungiform papillae were studied in insectivore hibernating bats (Pipistrellus pipistrellus and Rhinolophus ferrumequinum) in the various epochs of the year in order to show cell types the seasonal changes. On the basis of the ultrastructural aspects and the relationships with nerve endings, besides the few basal cells, three types of sensorial cells can be identified: dark type I cells, light type II cells and light type III cells. All gustatory cells, long and narrow shaped, extend from the epithelial basal lamina to the gustatory canal, where they send long microvillous expansions of the apical cytoplasm. These expansions, except those of the fungiform papillae which never extend beyond the lower two thirds, usually reach the external opening of the taste pore, always completely void of dense substance. Dark type I cells are characterized by a developed RER and large granules. Light type III cells show peculiar dense core light vesicles, labelled protein A-gold particles immunoreactive to 5-HT. These cells only in the foliate and fungiform papillae also have bundles of microtubules whose function is still unknown. After comparative evaluation and discussion of topographical and ultrastructural aspects with those of other mammals and humans, the Authors states that bats are provided with a valid gustatory system and that all cell types are involved in taste transduction.

Indomethacin-induced morphological changes in the rat gastric mucosa, with or without prior treatment with two proton pump inhibitors.

BACKGROUND: The mechanisms responsible for the gastric ulcerogenic effect of indomethacin are unclear. The importance of basal acid secretion on morphological changes by indomethacin was investigated. METHODS: Gastric lesions were macroscopically evaluated 6 h after indomethacin, 20 mg/kg intragastrically, in rats pre-treated with omeprazole (10-100 mumol/kg intragastrically) or lansoprazole (3-30 mumol/kg intragastrically). Glandular mucosa was processed for light, scanning and transmission electron microscopy 3 and 6 h after indomethacin in rats pre-treated with omeprazole (100 mumol/kg) or lansoprazole (30 mumol/kg). RESULTS: After 3 h, indomethacin caused extensive vasocongestion, oedema in the subepithelial region and superficial erosions. After 6 h, deeply extending focal necrosis involved 11% of the tissue. Leukocyte margination was occasionally seen at 3 h and consistently present at 6 h. Only at 6 h were endothelial cells altered. In rats pre-treated with omeprazole (100 mumol/kg) or lansoprazole (30 mumol/kg) grossly visible lesions were prevented. Oedema, erosions and necrosis were absent. Vasocongestion, vascular leakage and leukocytic margination were seen both at 3 and 6 h while no major damage of endothelial cells was observed. CONCLUSION: Indomethacin appears primarily to alter microcirculation, and microcirculation damage is dependent on acid for the progression to haemorrhagic lesions.

Gastroprotective activity of the novel proton pump inhibitor lansoprazole in the rat.

1. The protective activity of lansoprazole was evaluated on gastric mucosal lesions induced by intragastric 25% NaCl (1 ml/rat for 1 hr) and by indomethacin (20 mg/kg for 6 hr) in the rat and compared with that of omeprazole. 2. Lansoprazole (3, 10 and 10 mumol/kg i.g.) dose-dependently prevented the formation of indomethacin-induced lesions, the inhibition being 99% at the highest dose. Omeprazole, 10 mumol/kg i.g., enhanced the damage by indomethacin while higher doses caused a reduction, lesion index being reduced by 98% at 100 mumol/kg. 3. Histologically in lansoprazole- as well as in omeprazole-pretreated rats, indomethacin-induced necrosis of the mucosa was absent, luminal epithelium being intact. 4. Lansoprazole (30, 100 and 300 mumol/kg) and omeprazole (30, 100 and 300 mumol/kg) dose-dependently reduced the formation of lesions by hypertonic saline. 5. Present results indicated that lansoprazole and omeprazole protect the gastric mucosa in different experimental models of gastric ulceration.

[Structure and seasonal aspects of the bat gallbladder]. / Struttura ed aspetti stagionali della cistifellea di alcuni pipistrelli.

The gallbladder of the insectivorous hibernating bats (Pipistrellus pipistrellus et Rhinolophus ferrumequinum) in fullness, shows a fundus, a corpus and a neck. Its wall is formed by three layers: mucous membrane, with simple columnar epithelium on a thin basal lamina, beneath which is a "lamina propria" of connective tissue with blood and lymphatic vessels; lamina muscularis, constituted by three of four parallel bundles of smooth muscle cells, arranged in circular pattern; serous membrane of loose connective tissue, with precollecting lymphatic vessels and covered by mesothelial cells. By SEM the luminal surface of mucous membrane shows more or less prominent folds delimiting irregular areas with hexa-pentagonal outlines of the apical region of epithelial cells. This latter present a convex dome aspect with numerous microvilli, not changed significantly, during the different year periods. In summer and autumn the epithelial cells are characterized by developed organelles in supranuclear cytoplasm. During the prehibernating period, chiefly the cytoplasm shows multivesicular and heterogeneous dense bodies. In summer, autumn and initial hibernating period, epithelial cell cytoplasm presents microvilli with prominent "antennulae microvilares", numerous light micropinocytosis vesicles, little rough vesicles in apical portion. The mitochondria, grouping in supranuclear and basal cytoplasm, in lethargy, are characterized by small matrix osmiophilic granules. The epithelial cells are held together near the apical surface by tight and gap junctions, and some rare desmosomes only in Rhinolophus f.e. The cell membranes are deep pleated or interdigitate with wide intercellular spaces during the prehibernating period and experimental arousing, while in lethargy and summer months these present linear outline with scanty invaginations. The Authors compared these morphological aspects of bat gallbladder with other micromammalia's one, underline particularly the functional role of dense bodies, wide intercellular spaces and apical micropinocytosis vesicles. The different seasonal ultrastructural characteristics of epithelial cells are considered in close relation to an absorbing function connecting with balance of different metabolisms.

[Structure and immunohistochemistry of the hypophysis of Alosa fallax nilotica during the reproductive migratory cycle]. / Struttura ed immunoistochimica dell'ipofisi di Alosa fallax nilotica durante il ciclo migratorio per la riproduzione.

The Pituitary of the Alosa fallax nilotica consists of a pars distalis, a pars intermedia and a pars nervosa. The pars distalis is divided into rostral and proximal. The pars nervosa extends also into proximal pars distalis. The rostral pars distalis, on the contrary in primitive Teleostei, has organized into cellular cords defining cavities or intercommunicating follicles. These latter finish into the oral adenohypophysial duct, which don't open in pharyngeal cavity. The cords contain three cell types: the columnar cells LTH, TSH cells and basal cells not yet functionally determined. The cord cells are separated from follicle cavity by an one layer of flat degranulated cells. The LTH cell, the chief cell type, together the TSH and basal cells shows evident cytologic differences in organelles of synthesis and secretion during the various stages of the reproductive migration. Besides globular and baton-shape projections are underlined in the apical cytoplasm of LTH cells. These structure, extending into follicle cavity, seem to dispatch sensory function probably. The proximal pars distalis consists of compact cords of gonadotropic cells (particularly active during the stage of migration from rivers to the sea) with inserted orangiophilic cells (probably STH) and phloxine cells: these latter cells are chiefly arranged at the periphery and immunocytochemical positive for the antibodies against ACTH. The pars intermedia contains cord of ACTH cells, particularly manifest during the periods of transit from sea water to fresh water and on the contrary, they are characterized by positive-ACTH dense granules. On the ground of ultrastructural aspects of vesicles, the pars nervosa shows three types of nervous fibers which correlate with neurosecretory hypothalamus and catecholaminergic dopaminergic fibers. The Authors think of underlining particularly that the all cells of rostral pars distalis and chiefly the LTH cells are involved significantly in the regulation of reproduction and hydro-mineral metabolic equilibrium.

Comparative anatomical and ultrastructural features of the sensory papillae in the tongue of hibernating bats.

The papillae of the tongue dorsal surface of the insectivorous, hibernating bats (Vespertilionidae and Rhinolophidae), whose function is mainly sensorial, consist of two circumvallate papillae, two foliate papillae, located at the side edges at the glossopalatine arch, and numerous fungiform papillae. The circumvallate and foliate papillae are characterized not only by their position, but also by presence of several taste buds which open through the external orifice of the gustatory canal into the cavity of the vallum, or furrow, which divides the two folds of the lingual mucosa. The fungiform papillae (extremely numerous on the whole dorsal surface) are characterized by an unusual arrangement (along 3 oblique lines on the anterior two-thirds and predominantly on the middle line of the tongue body) and by the presence of only one to three taste buds which open on the heavily keratinized dorsal epithelial surface. The taste buds are made up of sensory cells with a light or dark matrix; their apical cytoplasmic expansions are not found beyond the middle part of the gustatory canal, in contrast with the circumvallate and foliate papillae which protrude from the orifice of the gustatory pore. Comparisons with the papillae of other types of bats and Insectivora and evaluations of the morphological characteristics and their functional values (unusual areas of distribution of the papillae, apical cytoplasmic expansions and behaviour of microfolds observed under SEM) have been made in different environmental conditions and nutritional habits, with attention to the mechanical events in the course of feeding.

Neuronal aspects and plasticity of inferior olivary complex and nucleus dentatus.

In primary or physiological ageing, the brain undergoes a 10% weight reduction. This datum was confirmed in vivo by radiological investigation and post mortem by morphological research (Grandi and Coll. 1990, 1990, 1991, 1991), that underscores the same reduction in the medulla oblongata and in the cerebellum. Morphological and cytohistological aspects of primary ageing have been considered for the inferior olivary nucleus and for the dentate nucleus in 41 male subjects, 32 belonging to the decades 7th, 8th, 9th, and 9 to the decades from the 3rd to the 6th. From the two latter structures, separated along standard cut surfaces, serial histological slices were carried out, and histochemical reactions were accomplished, together with immunohistochemistry, for synaptophysin, NF-2F11, NSE, S-100, GFAP, chromogranin. As the weight progressively decreased from the 7th to the 9th decade, it was noted neuronal loss, both in the inferior olivary nucleus, and in the dentate nucleus. Furthermore, the loss appeared to be followed by substitutive astroglial proliferation. Immunocytochemical reaction for synaptophysin revealed peculiar aspects. A consistently regular result of such method concerned the two nuclei studied, in primary ageing, thus allowing the interpretation of the perfect structure of synaptic vesicles as an indirect argument for neuronal plasticity.