Interactions of indole acetic acid with EGF and FSH in the culture of ovine preantral follicles

The mechanisms that regulate the gradual exit of ovarian follicles from the non-growing, primordial pool are very poorly understood. The objective of this study was to evaluate the effects of adding indole acetic acid (IAA), epidermal growth factor (EGF) and follicle stimulating hormone (FSH) to the media for in vitro culture of ovine ovarian fragments and determine their effects on growth activation and viability of preantral follicles. The ovarian cortex was divided into small fragments; one fragment was immediately fixed in Bouin (control). The other fragments were cultured for 2 or 6 days in culture plates with: minimum essential medium (MEM) supplemented with insulin-transferrin-selenium (ITS), pyruvate, glutamine, hypoxantine, bovine serum albumine and antibiotics (MEM+); MEM+ plus IAA (40 ng/mL); MEM+ plus EGF (100 ng/mL); MEM+ plus FSH (100 ng/mL); MEM+ plus IAA + EGF; MEM+ plus IAA + FSH; MEM+ plus EGF + FSH; or MEM+ plus IAA + EGF + FSH. After 2 or 6 days of culture in each treatment, the pieces of ovarian cortex were fixed in Bonin for histological examination. Follicles were classified as primordial or developing (primary and secondary) follicles. Compared to the control, in all media tested, the percentages of primordial follicles were reduced (P < 0.05) and the percentages of developing follicles were increased (P < 0.05) after 2 or 6 days of culture. Furthermore, culture of ovarian cortex for 6 days reduced the percentages of healthy, viable follicles when compared with the control (P < 0.05), except for cultures supplemented with IAA + EGF and EGF + FSH. In conclusion, the addition of IAA and EGF or EGF and FSH to the culture media were the most effective treatments to sustain the health and viability of activated ovine primordial follicles during in vitro culture. (c) 2005 Elsevier B.V. All rights reserved.

Flowering phenology, nectary structure and breeding system in Corymborkis flava (Spiranthoideae : Tropidieae), a terrestrial orchid from a Neotropical forest

Flowering phenology, breeding system and nectary structure of Corymborkis flava (Sw.) Kuntze were studied in a fragment of the Atlantic Forest in south-eastern Brazil. The flowering period extended from March (end of rainy season) to early June and seed dispersal occurred from June to September (dry season). Flowering peak occurred mainly in April, with up to 34 open flowers per plant being observed. The yellow, odourless and tubular flowers lasted similar to 7.8 days. The flowers present a perigonal nectary located in the basal lateral parts of the labellum; this is the first report on the nectary location and characterisation in the Tropidieae tribe. At the pre-anthesis stage, cells of both secretory parenchyma and epidermis of the nectary are filled with compound amyloplasts. However, starch grains were not observed in these tissues in senescent flowers, indicating that these starch grains are hydrolysed and used as source of sugars for nectar production. The nectar accumulates between the cuticle and the outer periclinal wall of the epidermal cells before. owing out into the nectar chamber. C. flava is a self-compatible species and spontaneous self-pollination does not occur because of hercogamy. The high pollinia removal (0.80) and insertion (0.82) per flower, as well as the high natural fruit-set indicate an efficient natural pollination system. The present study contributes for the knowledge of the diversity of reproductive strategies and nectary structures in Orchidaceae.

Placentation in the capybara (Hydrochaerus hydrochaeris), agouti (Dasyprocta aguti) and paca (Agouti paca)

Placentae of three hystricimorph rodents-capybara, agouti and paca-were examined by conventional histology, immunohistochemistry for cytokeratin and vimentin, and TUNEL staining. The placentae were divided into lobules of labyrinthine syncytium separated by interlobular and marginal trophoblast. The subplacenta comprised cytotrophoblasts, supported on lamellae of allantoic mesoderm, and syncytiotrophoblast. The central excavation was still apparent in the definitive placenta of capybara. In agouti and paca, the decidua of the junctional zone formed a mesoplacenta comprising a capsule and a pedicle. Towards term the pedicle formed a tenuous attachment between placenta and uterine wall comprising a few maternal vessels surrounded by degraded tissue. In paca placenta, it was shown by TUNEL staining that breakdown of this tissue occurred by apoptosis. The visceral yolk sac was highly villous and, in agouti, the yolk sac villi were extremely long. Lateral to its attachment to the placenta, the fetal surface was covered with non-vascular yolk sac endoderm. A layer of spongiotrophoblast cells was interposed between the endoderm and the marginal trophoblast. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

First record of petrified Permian pecopterids from the Parana Basin, Brazil (Corumbatai Formation, Passa Dois Group, northeastern State of São Paulo): Morphology, anatomy and paleoecological implications

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Activity of the Ovarian Germinal Epithelium in the Freshwater Catfish, Pimelodus maculatus (Teleostei: Ostariophysi: Siluriformes): Germline Cysts, Follicle Formation and Oocyte Development

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Histopathological changes induced by extracts from the tissue covering the stingers of Potamotrygon falkneri freshwater stingrays

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

FLORAL ANATOMY of XYRIDS (POALES): CONTRIBUTIONS TO THEIR REPRODUCTIVE BIOLOGY, TAXONOMY, and PHYLOGENY

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Fine structure of lamellated nerve endings in the gingiva of man and the Cebus apella monkey

Gingival mucosae of man and the adult Cebus apella monkey were fixed for 3 hr in modified Karnovsky fixative containing 2.5% glutaraldehyde, 2% formaldehyde in 0.1 M sodium phosphate buffer (pH=7.4). The specimens were postfixed in 1% osmium tetroxide in 0.1 M sodium phosphate buffer at 4°C for 2 hr, dehydrated in a graded alcohol series and embedded in Epon 812. Thick sections of 1-3 μm and ultrathin sections of 40-80 nm in thickness were cut with glass knives on an LKB ultramicrotome. The thick sections were stained with toluidine blue solution, and the grids were stained with uranyl acetate and lead citrate and examined under a Philips EM-301 electron microscope. Our observations permitted us to conclude that: both gingival mucosae, of man and the Cebus apella monkey, have lamellar nerve endings; these corpuscles are localized in the papillar space of the epithelium and do not contact closely with the basement membrane; the nerve endings are composed of an afferent fiber which subdivides several times and forms irregular flattened or discoidal expansions; the laminae of the lamellar cells are very thin near the terminal axon and are larger and irregular in shape at the peripheral portion of the corpuscle; the terminal axon shows abundant mitochondria, myelin figures, clear vesicles, and multivesicular bodies; between the axoplasm membrane and adjacent cytoplasmic lamina and between the lamellae, small desmosome type junctions are noted; and the cytoplasmic material of the lamellae cells is characterized by the presence of numerous microfilaments, microtubules, mitochondria, rough endoplasmic reticulum, and caveolae.

Submicroscopical features of leaves of xyris species

The leaf infrastructure of five Xyris species were examined using scanning electron microscope (SEM), transmission electron microscope (TEM) and histochemical methods. All studied leaves show some features in epidermis and mesophyll, which were of considerable adaptative significance to drought stress. Such features included the occurrence of a pectic layer on the stomatal guard cells and the presence of a nehvork ofpectic compounds in the cuticle. Pectic compunds were also in abundance in lamellated walls of the mesophyll cells and on the inner surface of the sclerified cell walls of the vascular bundle sheaths. There were also specialized chlorenchymatous peg cells in the mesophyll and drops of phenolic compounds inside the epidermal cells.

The ultrastructure of the guinea pig rete testis

The chambers of the rete testis (RT) of guinea pig are lined by a simple epithelium, whose cells are squamous, cubical and columnar in shape. The epithelial cells with distinct shapes were counted and the quantitative analysis of the number of these cells showed relative predominance of cubical cells. The ultrastructural observations showed predominance of membrane interdigitations among the epithelial cells. These cells present common cytoplasmic organelles. The Golgi complex polarity is typical with observation of electronlucent vesicles on the Golgi cis face closely related to rough endoplasmic reticulum (ER) lamellae, mitochondria and large number of polysomes on the Golgi trans face. These related structures present in Golgi area of RT cells suggest secretory activity which maybe occurs in the RT epithelium. Endocytotic process also occurs in the RT and this function probably concerns the uptake of substances and resorption of seminiferous fluid. Apical cilia present in RT epithelium cells are related with fluid transport and perhaps with chemoreception. Presence of spermatozoa portions enclosed into the cytoplasm of some epithelium cells has been refferred to as spermatophagy. The RT complex is mainly supported by loose connective tissue, with collagen fibres and some Leydig cells. Leydig cells are adjacent to the network channels of the septal part of the RT and apparently are able to secrete inside the RT lumen.

Características anatômicas de espinheira-santa (Maytenus ilicifolia Mart. ex Reissek e Maytenus aquifolia Mart.) e mata-olho (Sorocea bonplandii (Baill.) Burg. Lanj. & Boer.) para o controle de qualidade da matéria prima

Maytenus ilicifolia and Maytenus aquifolia (Celastraceae) both designated espinheira-santa have proven anti-ulcer activity. Morphologic similarities between leaves of espinheira-santa and mata-olho (Sorocea bonplandii), has motivated fakes in the market of phytotherapy. The present work consisted of the anatomical study, including stem and leaf, of the species M. ilicifolia, M. aquifolia and S. bomplandii. Samples of adult leaves and stem of plants located in the cities of Maringá and Marialva were collected. Both are located in the northwest region of Paraná State. The botanical material was prepared with using usual techniques of anatomy. The leaves of both Maytenus species presented great similarities, characterizing itself for the presence of epidermal cells with straight walls, biseriate palisade tissue, petiole vascular system represented by unique amphicribal bundle and sclereids, which were present in the stems of these two species. S. bomplandii leaves differed of Maytenus species for presenting epidermal cells with undulated walls, uniserite palisade tissue, petiole vascular system represented by many collateral bundles and gelatinous fibres. Non-glandular trichomes, glandular trichomes, and laticifer only occur in S. bomplandii.

Características estruturais da parede das artérias renal e femoral de coelho (Oryctolagus cuniculus)

In the renal and femoral arteries of rabbit was verified that both vessels had walls structured by myostromal components, despite of their different distributive of blood, being the renal artery a visceral blood vessel and the femoral artery a parietal vessel. This wall pattern in these vessels concerned to presence of connective stromal elements (collagen and elastic fibres and lamellae) and smooth muscle cells coexisting with some equilibrium in the wall structure of the renal and femoral arteries, mainly in the medial layer architecture. An intimal folding pattern was verified around the vascular lumen, possibly related to capacitance of the both arteries regarding to variability of pressure levels in cardiac cycle. Furthermore, myostromal relations of connective elements and smooth muscle cells verified in the medial layer and the network formed by connective elements in the adventitial layer of these arteries contributed to maintenance of wall viscoelasticity properties of the vessels.

Cyclosporin but not tacrolimus significantly increases salivary cytokine contents in rats

Background: Cyclosporin (CsA) and tacrolimus (FK-506) are immunosuppressive drugs that specifically inhibit T-cell activation via calcineurin inhibition. Gingival overgrowth is a common side effect following the administration of CsA. The severity of gingival overgrowth seen in patients taking FK-506 is less than that observed with CsA. Little is known about the involvement of saliva in drug-induced gingival overgrowth. The purpose of this study was to investigate the salivary contents of tumor growth factor β1 (TGF-β1), epidermal growth factor (EGF), and interleukin-6 (IL-6) as well as the hystometry of gingival tissue obtained from rats treated with either FK-506 or CsA. Methods: For 30 or 60 days rats received daily subcutaneous injection doses of either CsA or FK-506 (10 mg/kg). The concentrations of TGF-β1, EGF, and IL-6 in saliva were determined by enzyme-linked immunosorbent assay, and after histological processing, the oral epithelium and connective tissue were assessed at the region of the lower first molars. Results: The levels of TGF-β1, EGF, and IL-6 in saliva were not significantly altered by any of the treatments after 30 days. After 60 days of treatment with CsA, gingival overgrowth and significant increase in salivary TGF-β1, EGF, and IL-6 concentrations were observed; no statistically significant changes were induced by FK-506. Conclusion: Within the limits of this experimental study, it can be concluded that CsA, but not FK-506, induced gingival overgrowth associated with an increase of the salivary levels of the cytokines TGF-β1, EGF, and IL-6.

Additional observations on the gross morphology and microstructure of Baccaconularia Hughes, Gunderson et Weedon, 2000, a Cambrian (Furongian) conulariid from the north-central USA

Baccaconularia Hughes, Gunderson et Weedon, 2000, from the Furongian Series (Cambrian System) of the north-central USA, has been interpreted as a conulariid cnidarian, based on a suite of gross morphological similarities shared only with other post-Cambrian genera currently assigned to this group. Closely spaced, squarish to subrectangular facial nodes of Baccaconularia are aligned in distinct longitudinal files. Nodes also display a subtler, more or less rectilinear transverse alignment, though this pattern commonly is disrupted by offset parallel to the longitudinal files. In their shape and pattern of arrangement, the nodes of Baccaconularia are most similar to the squarish to elongate nodes of Pseudoconularia Bouček, 1939. Longitudinal node files of Baccaconularia may also be compared with the longitudinal facial ridges of Conularia cambria Walcott, 1890 from the Furongian of Wisconsin. Apical angles of Baccaconularia range from approximately 13° to 14.5°. Scanning electron imaging of B. cf. robinsoni shows that its thin, phosphatic skeleton is finely lamellar, with the thickness of individual lamellae measuring approximately 1 μm. The skeleton also exhibits microscopic circular pores and crater-like pits that range from approximately 5 to 10 μm in diameter. These pores and pits are similar in size, geometry, areal density and pattern of arrangement to those of many post-Cambrian conulariids. Microscopic circular pores are documented here for the first time in the genus Archaeoconularia Bouček, 1939 from the Upper Ordovician of the Czech Republic. Although the origin of the pores and pits is open to alternative interpretations, the discovery of these features and fine lamination in Baccaconularia strengthens the argument that this genus is a Cambrian conulariid. © 2006 Nanjing Institute of Geology and Palaeontology, CAS.

Morfo-anatomia do eixo vegetativo aéreo de Achyrocline alata (Kunth) DC. (Asteraceae)

Achyrocline alata occurs in a dense and low vegetation in Mato Grosso do Sul State, Brazil and it has been studied under physical-chemical and yield aspects, being scarce anatomical data. The present work has as objective the study of the aerial vegetative axis. Stem is cylindrical and hairy, having five wings, which are arisen from their leaves. The blade shows the presence of uniseriate epidermis, which is covered by the cuticle of variable thickness; and non-glandular and glandular trichomes. The non-glandular trichomes are uniseriate and multicellular and have their apex cell in whip form, while the glandular trichomes are multicellular, uniseriate or biseriate. The mesophyll is dorsiventral with uni-stratified palisade parenchyma in most of the cases and lacunal parenchyma formed by three to four layers of irregular cells. Only one collateral vascular bundle occurs in the midvein. Stem in transversal section is covered by epidermis with trichomes similar to leaves; the cortex is constituted by a discontinuous area of angular collenchyma, which is followed by chlorophyll parenchyma. Vascular cylinder that is delimited by pericycle shows vascular bundles of collateral type. In the secondary structure, the periderm is originated from epidermal and subepidermal tissues. In vascular region, the fascicular cambium differs into secondary xylem and phloem, while interfascicular cambium produces sclerenchymatous tissue.