Effect of environmental mycobacteria (Mycobacterium avium) on immunity induced by a DNA vaccine (DNAhsp65) against tuberculosis

The efficacy of BCG vaccine (attenuated Mycobacterium bovis) against pulmonary tuberculosis varies enormously among different populations. The prevailing hypothesis attributes this variation to interactions between the vaccine and mycobacteria common in the environment. Studies have revealed that most protective antigens expressed by the antituberculous vaccine are conserved in M. avium, supporting the hypothesis that exposure to environmental mycobacteria generates a cross-reactive immune response that interferes with BCG efficacy. In this study we investigated the effect of a prior exposure to heat-killed M. avium on the immune response and the protective efficacy induced by a genetic vaccine pVAXhsp65 (hsp65 gene from M. leprae inserted in pVAX vector) against experimental tuberculosis. To evaluate the effect on the immune response, female BALB/c mice were initially injected with distinct doses (0.08×106, 4×106, and 200×10 6) of heat-killed M. avium by subcutaneous route. Three weeks later, the animals were immunized with 3 doses of DNAhsp65 by intramuscular route (100μg/15 days apart). Control groups received only M. avium, vaccine (pVAXhsp65), vector (pVAX) or saline solution. Cytokine production and antibody levels were determined by ELISA. To evaluate the effect on the protective efficacy, animals were initially sensitized with 200×106 heat-killed CFU of M. avium by subcutaneous route and then immunized with 3 doses of pVAXhsp65 (100μg/15 days apart) by intramuscular route. Control groups were injected with saline, pVAX (4 doses), pVAXhsp65 (4 doses), M. avium or M. avium plus pVAX (3 doses). Fifteen days after last DNA dose, the animals were infected with 1×104 viable CFU of H37Rv M. tuberculosis by intratracheal route. Thirty days after challenge, the animals were sacrificed and the bacterial burden was determined by counting the number of CFU in the lungs. Lung histological sections were also analyzed. Splenic cells from primed animals produced more IL-5 but less IFN-gamma than non-primed ones. Also, prior contact with M. avium determined higher production of IgG1 and IgG2a anti-hsp65 antibodies in comparison to control groups. However, this higher immune response did not decrease the bacterial burden in the lungs. In addition, prior sensitization with M. avium decreased the parenchyma preservation observed in the group immunized only with pVaxhsp65. These results indicate that environmental mycobacteria can interfere with immunity and protective efficacy induced by DNAhsp65.

Estudo morfológico do tecido hepático de Leporinus macrocephalus

The objective of this work was to analyze the liver of Leporinus macrocephalus at the macroscopic and histologic level by evaluating the characteristics of the hepatic tissue. The fishes from a fishery in Bauru, S.P., were collected and sacrificed by destroying the spinal cord; afterwards the collected hepatic tissue was fixed and followed by histological routine. Macroscopic analysis of L. macrocephalus liver showed an organ of homogeneous structure, with red-brown color, located in the medial region of the body, caudal to heart and after the gills. It showed 3 pyramidal lobes, one central and two lateral right and left. The histological analysis showed the hepatic tissue constituted by hepatocytes cords surrounded by sinusoids, named muralium duplex. It yet revealed the diffused, distribution bile system by hepatic parenchyma, showing ducts with simple cubic epithelium, surrounded by muscular fibres and connective tissue. Cells of the exocrine pancreatic tissue were observed surrounding blood vessels in acinar arrangement as zymogen granules. Furthermore, there are melanomacrophages centers distributed along the hepatic parenchyma, preferably next to the blood vessels, constituted by cells accumulating material, such as melanin and lipofucsin, whose presence may be related to the nutritional status of the fish.

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.

Morfogênese do testículo de embriões e fetos de vacas da raça nelore (Bos taurus indicus)

The aim of this study was to accompany the process of testicular development from the non-differentiable phase to its complete formation. Embryos and fetuses of Nelore breed cows (Bos Taurus indicus) were obtained in slaughterhouses near the Uberlandia city, Minas Gerais. The gonads and the embryos were fixed in Bouin's fixative and afterwards processed for conventional optical microscopy. The gonadal was observed firstly in a 1.0 cm long embryo. In 2.5 cm long embryos the presence of the albuginea allows the sex identification. The mean thickness of the albuginea ranged from 29.08 to 558.45 mm. Gradually increase of vascularization of the albuginea and parenchyma is observed. The mediastinum is located centrally. There was a decrease in the space occupied by the testicular cords, from 63.71 to 41.99% of the total testes volume. Its diameter ranged from 31.68 to 48.80 mm. The diameter of germinal cells (and their nuclei) was from 12.27 (6.65) to 16.95 914.21) mm. The quantity of germinal cells by cross section of cord decreased from a maximum of 2.80 to 0.76. The total number of germinal cells was from 16 at the beginning of colonization of the gonad to 18.32 x 106 at the end of the study. The number of Sertoli's cells by cross section of cord ranged from 10.00 to 16.25. The results obtained show that the origin and formation of testes in embryos and fetuses from Nelore breed cows (Bos taurus indicus) does occur in a very similar way to what is described for Bos taurus taurus.

rastreamento histoquímico, fitoquímico e biológico de folhas de Plinia cauliflora (DC.) Kausel (Myrtaceae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Female sex hormones mediate the allergic lung reaction by regulating the release of inflammatory mediators and the expression of lung E-selectin in rats

Background: Fluctuations of estradiol and progesterone levels caused by the menstrual cycle worsen asthma symptoms. Conflicting data are reported in literature regarding pro and anti-inflammatory properties of estradiol and progesterone.Methods: Female Wistar rats were ovalbumin (OVA) sensitized 1 day after resection of the ovaries (OVx). Control group consisted of sensitized-rats with intact ovaries (Sham-OVx). Allergic challenge was performed by aerosol (OVA 1%, 15 min) two weeks later. Twenty four hours after challenge, BAL, bone marrow and total blood cells were counted. Lung tissues were used as explants, for expontaneous cytokine secretion in vitro or for immunostaining of E-selectin.Results: We observed an exacerbated cell recruitment into the lungs of OVx rats, reduced blood leukocytes counting and increased the number of bone marrow cells. Estradiol-treated OVx allergic rats reduced, and those treated with progesterone increased, respectively, the number of cells in the BAL and bone marrow. Lungs of OVx allergic rats significantly increased the E-selectin expression, an effect prevented by estradiol but not by progesterone treatment. Systemically, estradiol treatment increased the number of peripheral blood leukocytes in OVx allergic rats when compared to non treated-OVx allergic rats. Cultured-BAL cells of OVx allergic rats released elevated amounts of LTB4 and nitrites while bone marrow cells increased the release of TNF-α and nitrites. Estradiol treatment of OVx allergic rats was associated with a decreased release of TNF-α, IL-10, LTB4 and nitrites by bone marrow cells incubates. In contrast, estradiol caused an increase in IL-10 and NO release by cultured-BAL cells. Progesterone significantly increased TNF- α by cultured BAL cells and bone marrow cells.Conclusions: Data presented here suggest that upon hormonal oscillations the immune sensitization might trigger an allergic lung inflammation whose phenotype is under control of estradiol. Our data could contribute to the understanding of the protective role of estradiol in some cases of asthma symptoms in fertile ans post-menopausal women clinically observed. © 2010 de Oliveira et al; licensee BioMed Central Ltd.

Intoxicação por chumbo em um cão - relato de caso

The present study reports the occurrence of lead poisoning in a male, 2 year-old mongrel dog attended at UNESP Botucatu Veterinary Hospital. The animal was brought to the hospital with claim of gastrointestinal and nervous disturbs for 10 days, worsening in the last 24 hours. After unsuccessfully attempts to control status epileticus, the animal died. Hematological evaluation revealed significant rubricytosis and basophilic stippling, highly suggestive of lead poisoning. Serum lead concentration confirmed toxic levels of this element. Necropsy evidenced the presence of foreign bodies in stomach (one stone and two metallic objects) and, microscopically, characteristic inclusions in renal parenchyma. Fragments obtained from liver and kidneys revealed lead concentrations of 0, 8 μg/g and 25 μg/g, respectively, which in association to clinical signs of saturnism, determined the diagnosis of lead poisoning.

Morphological aspects of the liver of the Podocnemis expansa (Testudines, podocnemididae)

The liver of P. expansa was characterized morphohistologically. To this end, twenty livers from clinically healthy male and female Podocnemis expansa, weighing from 2.0 to 4,5 kg, supplied by the commercial breeder Fazenda Moenda da Serra, in Araguapaz, state of Goiás, Brazil, were analyzed macro-and microscopically. The coelomatic cavity was opened and the topography of the fresh organs was examined visually. After the histological preparation, the slides were stained with Hematoxylin and Eosin (HE), Periodic Acid-Schiff (PAS), Gomori Trichrome, Reticulin and Picrosirius. The liver of P. expansa is a voluminous organ with an approximately rectangular shape and brown coloration, varying from light to dark shades, and is divided into a right lobe, left lobe, and a central portion. The right lobe is the largest of the three portions. The gall bladder is located in a depression in the caudal portion of the right lobe, where the gall duct begins and empties into the duodenum. Histologically, the hepatocytes are arranged in the form of double cords surrounded by winding sinusoidal capillaries. In cross section, they resemble acini containing approximately two to five hepatocytes surrounding a probable central biliary canaliculus. The hepatocytes are polyhedral or pyramidal in shape, of uniform size, with a few central nuclei and others displaced peripherally, and the cytoplasm is little eosinophilic when analyzed by the HE staining technique. The parenchyma is supported by delicate reticular fibers surrounding hepatocytes and sinusoids. The parenchyma and perisinusoidal spaces contain large quantities of melanomacrophages, mainly close to the portal spaces.

Anatomy of the floral nectaries of some neotropical Salacioideae (Celastraceae)

Floral nectaries have contributed to the systematics of different taxonomic groups. Since those of the neotropical genera included in subfamily Salacioideae-Cheiloclinium Miers, Peritassa Miers, Salacia L. and Tontelea Aubl.-have different forms and positions, we explored their anatomy to delimit more precisely the genera of subfamily Salacioideae. Buds and open flowers of six species were treated following the usual techniques in plant anatomy. The obtained data were helpful in characterizing the floral nectary anatomy of the studied species. Furthermore, some features such as form, position and surface of nectaries; form of their epidermal cells; presence and distribution of stomata; occurrence of idioblasts containing druses in the nectariferous parenchyma; and absence of nectary vascularization can contribute to the taxonomy and phylogeny of the Salacioideae studied. In most of the studied species the nectar is probably released by both the stomata and the nectary epidermal surface. In Cheiloclinium cognatum, the structure acknowledged as nectary is actually a vestigial tissue and the functions of attracting and rewarding pollinators has phylogenetically migrated to the stigmatic region. The druses and phenolic substances observed in the nectariferous parenchyma probably help defend flowers against herbivore attacks. The minute size of the nectaries of Salacioideae may explain the absence of vascularization. The floral nectaries of Salacia elliptica are epithelial while those of the other species are mesenchymal. © 2012 Springer-Verlag Wien.

Structural changes in soybean seed coat due to harvest time and storage

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

Annexin-A1 peptide down-regulates the leukocyte recruitment and up-regulates interleukin-10 release into lung after intestinal ischemia-reperfusion in mice

Background: Intestinal ischemia/reperfusion (IR) injury is a serious and triggering event in the development of remote organ dysfunction, from which the lung is the main target. This condition is characterized by intense neutrophil recruitment, increased microvascular permeability. Intestinal IR is also responsible for induction of adult respiratory distress syndrome, the most serious and life-threatening form of acute lung injury. The purpose of this study was to investigate the effect of annexin-A1 protein as an endogenous regulator of the organ remote injury induced by intestinal ischemia/reperfusion. Male C57bl/6 mice were subjected to intestinal ischemia, induced by 45 min occlusion of the superior mesenteric artery, followed by reperfusion. Results: The intestinal ischemia/reperfusion evoked a high intensity lung inflammation as indicated by the number of neutrophils as compared to control group. Treatment with annexin-A1 peptidomimetic Ac2-26, reduced the number of neutrophils in the lung tissue and increased its number in the blood vessels, which suggests a regulatory effect of the peptide Ac2-26 in the neutrophil migration. Moreover, the peptide Ac2-26 treatment was associated with higher levels of plasma IL-10. Conclusion: Our data suggest that the annexin-A1 peptidomimetic Ac2-26 treatment has a regulatory and protective effect in the intestinal ischemia/reperfusion by attenuation of the leukocyte migration to the lung and induction of the anti-inflammatory cytokine IL-10 release into the plasma. The anti-inflammatory action of annexin-A1 and its peptidomimetic described here may serve as a basis for future therapeutic approach in mitigating inflammatory processes due to intestinal ischemia/reperfusion. © 2013 Guido et al.; licensee BioMed Central Ltd.

A computer vision approach to quantify leaf anatomical plasticity: A case study on gochnatia polymorpha (less.) cabrera

Inferences about leaf anatomical characteristics had largely been made by manually measuring diverse leaf regions, such as cuticle, epidermis and parenchyma to evaluate differences caused by environmental variables. Here we tested an approach for data acquisition and analysis in ecological quantitative leaf anatomy studies based on computer vision and pattern recognition methods. A case study was conducted on Gochnatia polymorpha (Less.) Cabrera (Asteraceae), a Neotropical savanna tree species that has high phenotypic plasticity. We obtained digital images of cross-sections of its leaves developed under different light conditions (sun vs. shade), different seasons (dry vs. wet) and in different soil types (oxysoil vs. hydromorphic soil), and analyzed several visual attributes, such as color, texture and tissues thickness in a perpendicular plane from microscopic images. The experimental results demonstrated that computational analysis is capable of distinguishing anatomical alterations in microscope images obtained from individuals growing in different environmental conditions. The methods presented here offer an alternative way to determine leaf anatomical differences. © 2013 Elsevier B.V.

Macrophage migration inhibitory factor and oral cancer

Background: Macrophage migration inhibitory factor (MIF) is a pleiotropic cytokine with pro-inflammatory functions and involved in tumorigenesis. The aim of this study was to evaluate the expression and localization of the macrophage MIF in oral squamous carcinoma (OSC). In addition, the relationship between MIF expression and clinicopathological parameters such as survival data, tobacco use, alcohol habits, TNM stage, tumor graduation, and peritumoral inflammatory infiltrate were evaluated. Methods: Using immunohistochemistry, expression and localization of MIF was detected in 44 specimens of OSC. The absolute number and relative proportions of MIF-positive cells detected were also determined separately for tumor parenchyma vs. stroma. All counts were determined from 10 consecutive high-power fields using an integration graticule. Moreover, some parameters were analyzed separately for lip and intra-oral cancers. Results: Migration inhibitory factor-positive cells were observed in both the tumor parenchyma and in inflammatory cells of all specimens. In contrast, MIF expression was not detected in tumoral nests associated with poorly differentiated tumors. In specimens of lip cancer, a greater number of MIF-positive stromal immune cells were detected than in intra-oral cancer specimens (Mann-Whitney test, P = 0.049). Conclusions: Oral squamous carcinoma cells consistently express MIF independent of their location. Lip tumors presented more MIF-positive peritumoral inflammatory cells, similar to control, suggesting that immunological differences in leukocyte activation exist between in lip and intra-oral cancers. © 2012 John Wiley & Sons A/S.

Photosynthetic and anatomical responses of Eucalyptus grandis leaves to potassium and sodium supply in a field experiment

Although vast areas in tropical regions have weathered soils with low potassium (K) levels, little is known about the effects of K supply on the photosynthetic physiology of trees. This study assessed the effects of K and sodium (Na) supply on the diffusional and biochemical limitations to photosynthesis in Eucalyptus grandis leaves. A field experiment comparing treatments receiving K (+K) or Na (+Na) with a control treatment (C) was set up in a K-deficient soil. The net CO2 assimilation rates were twice as high in +K and 1.6 times higher in +Na than in the C as a result of lower stomatal and mesophyll resistance to CO2 diffusion and higher photosynthetic capacity. The starch content was higher and soluble sugar was lower in +K than in C and +Na, suggesting that K starvation disturbed carbon storage and transport. The specific leaf area, leaf thickness, parenchyma thickness, stomatal size and intercellular air spaces increased in +K and +Na compared to C. Nitrogen and chlorophyll concentrations were also higher in +K and +Na than in C. These results suggest a strong relationship between the K and Na supply to E. grandis trees and the functional and structural limitations to CO2 assimilation rates. © 2013 John Wiley & Sons Ltd.

Anatomy of scapes, bracts, and leaves of Paepalanthus sect. Diphyomene (Eriocaulaceae, Poales) and its taxonomic implications

Paepalanthus sect. Diphyomene comprises 18 species with a convoluted taxonomic history. Aiming to correlate anatomical structures with the systematics of this group and its relatives, we studied the anatomy of scapes, reproductive axis bracts, and leaves of 20 Paepalanthus species. Bracts and leaves show differences in epidermal cell thickening; mesophyll width; vascular bundle arrangement; presence or absence of a hypodermis; types of cells in the vascular bundle sheath extensions; margin shape and composition; and presence or absence of aquiferous parenchyma. Scapes differ in contour, rib number, and pith size. Some diagnostic characters found are presence of aquiferous parenchyma and absence of vascular bundle sheath extensions in leaves of P. urbanianus; vascular bundles decreasing in size towards the margin of leaves and bracts, and scapes with a triangular contour in P. flaccidus; scapes with nine ribs in P. acanthophyllus and ten in P. macer. All anatomical features are summarized in tables. These results aid in the identification and characterization of the species of P. sect. Diphyomene. They also support the current section circumscription, reinforcing the relevance of the anatomical characters in order to define natural groups. © 2012 The New York Botanical Garden.