Sequential morphological characteristics of murine fetal liver hematopoietic microenvironment in Swiss Webster mice

Embryonic hematopoiesis occurs via dynamic development with cells migrating into various organs. Fetal liver is the main hematopoietic organ responsible for hematopoietic cell expansion during embryologic development. We describe the morphological sequential characteristics of murine fetal liver niches that favor the settlement and migration of hematopoietic cells from 12 days post-coitum (dpc) to 0 day post-partum. Liver sections were stained with hematoxylin and eosin, Lennert’s Giemsa, Sirius Red pH 10.2, Gomori’s Reticulin, and Periodic Acid Schiff/Alcian Blue pH 1.0 and pH 2.5 and were analyzed by bright-field microscopy. Indirect imunohistochemistry for fibronectin, matrix metalloproteinase-1 (MMP-1), and MMP-9 and histochemistry for naphthol AS-D chloroacetate esterase (NCAE) were analyzed by confocal microscopy. The results showed that fibronectin was related to the promotion of hepatocyte and trabecular differentiation; reticular fibers did not appear to participate in fetal hematopoiesis but contributed to the physical support of the liver after 18 dpc. During the immature phase, hepatocytes acted as the fundamental stroma for the erythroid lineage. The appearance of myeloid cells in the liver was related to perivascular and subcapsular collagen, and NCAE preceded MMP-1 expression in neutrophils, an occurrence that appeared to contribute to their liver evasion. Thus, the murine fetal liver during ontogenesis shows two different phases: one immature and mainly endodermic (<14 dpc) and the other more developed (endodermic-mesenchymal; >15 dpc) with the maturation of hepatocytes, a better definition of trabecular pattern, and an increase in the connective tissue in the capsule, portal spaces, and liver parenchyma. The decrease of hepatic hematopoiesis (migration) coincides with hepatic maturation

Estudo histopatológico e molecular de embriões de Gallus gallus domesticus (Linnaeus, 1758) infectados com o vírus da Febre Amarela 17DD

Made available in DSpace on 2016-05-11T13:01:04Z (GMT). No. of bitstreams: 2 pedro_manso_ioc_dout_2014.pdf: 3405057 bytes, checksum: 6746c4f31f57631cda4cf25c55563336 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2014Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, BrasilA vacina contra febre amarela é produzida a partir da inoculação do vírus atenuado 17DD em ovos embrionados de galinha. Esta vacina é extremamente eficaz e segura, gerando imunidade que pode perdurar por até trinta e cinco anos. Embora a replicação deste vírus em embriões de galinha seja utilizada desde 1937, pouco se sabe sobre os aspectos da infecção nestes embriões, especialmente que órgãos, tecidos e células são responsáveis pela replicação viral. Nesse trabalho, analisamos embriões de galinha (Gallus gallus) infectados pelo vírus da FA 17DD em diferentes tempos de infecção (24, 48, 72 e 96 horas) conforme as condições empregadas na produção de vacina na Fiocruz. Identificamos o vírus da Febre Amarela através de imunofluorescência em diferentes tecidos, correlacionamos a presença deste agente infeccioso às pequenas reações histopatológicas observadas nos tecidos, validamos essa detecção pela confirmação do material genético viral e seu sequenciamento, e confirmamos que este vírus replica na região onde foi identificado pela presença detecção de seu intermediário replicativo. Nesse sentido observamos que as alterações histopatológicas que ocorrem nos embriões de galinha infectados pelo vírus FA 17DD se apresenta branda e sistêmica ao longo do tempo analisado. Nossos dados apontam que as primeiras células a manifestar a infecção são mioblastos com aspecto mesenquimal que puderam ser observados no coração e no músculo esquelético a partir de 48 horas de infecção Após 72 horas, o vírus FA 17DD replica em células do músculo esquelético, cardiomiócitos, células da glia e neurônios, no epitélio tubular renal, parênquima pulmonar e fibroblastos. Nossos dados permitem sugerir o tecido muscular esquelético como um local privilegiado na produção das partículas virais. Após 96 horas a infecção se torna mais intensa no sistema nervoso e se mantém nos mesmos níveis nos demais tecidos já infectados. O conjunto de dados gerados nesse trabalho contribui para elucidar aspectos importantes sobre a patologia da febre amarela em embriões de galinha, e evidenciar os tecidos e células responsáveis pela produção do vírus FA 17DD nestes embriões. Estes dados podem ser úteis na compreensão e formulação de novas estratégias de produção da vacina, além de impactar no desenvolvimento de estratégias baseadas no uso do vírus FA 17DD como plataforma de produção para outras vacinasYellow fever vaccine is produced from the inoculation of attenuated virus YF 17DD in embryonated chicken eggs. This vaccine is extremely effective and safe, generating immunity that can persist across up to thirty-five years. Although replication of this virus in chicken embryos is used since 1937, little is known about aspects of infection in these embryos, especially that organs, tissues and cells are responsible for viral replication. In this study we analyzed chicken embryos (Gallus gallus) infected in vaccine production (Biomanguinhos) with YF 17DD virus in different times post infection (24, 48, 72 and 96 hours). Here it was possible to detect the Yellow Fever Virus by immunofluorescence, to correlate this presence with tiny tissue reactions, to validate it by genomic RNA detection and to sequence it in the same studied area, confirming that this virus is replicated in these regions by the replicative intermediate detection. In this thesis the histopathological changes that occur in chicken embryos infected by YF 17DD virus during the production of yellow fever vaccine were observed in a kinetic way. We observed that the infection in these embryos presented itself mild and systemic. Our data show that the first cells which express infection are myoblasts with mesenchymal shape that could be observed in the heart and skeletal muscle at 48 hours of infection. After 72 hours the yellow fever virus 17DD replicates mainly in skeletal muscle cells, cardiomyocytes, glial cells and neurons, but also in the renal tubular epithelium, lung parenchyma and fibroblasts. Our findings suggested skeletal muscle tissue as a main place in the production of viral particles. After 96 hours the infection becomes more intense in the nervous system and is maintained at the same levels in other tissues already infected. The data generated in this study contributes to elucidate important aspects of the yellow fever pathology in chicken embryos, and elucidate the tissues and cells responsible for YF 17DD virus production in this model. Our data may be helpful in the understanding and design new strategies of vaccine production, and impact in development of strategies based on the use of the virus YF 17DD as a platform for other vaccines production

Morphological and morphometric study of pre-ovigerous and post-ovigerous adults of Tanaisia (Paratanaisia) bragai (Santos, 1934) (Digenea, Eucotylidae)

The aim of this study was to obtain data on the morphology and morphometry of pre-ovigerous and post-ovigerous adults of the species Tanaisia (Paratanaisia) bragai, using confocal laser scanning microscopy to obtain tomographic images of the suckers and tegument. For morphometric analysis, 45 specimens (30 pre-ovigerous adults and 15 post-ovigerous adults) were measured with the aid of an ocular micrometer coupled to the objective of a photonic microscope. Pre-ovigerous and post-ovigerous adult individuals, stained with Mair carmalumen and mounted in permanent preparations, were analyzed by means of confocal laser scanning microscopy. Positive correlation was detected between the body length and ovary length of post-ovigerous adults (rs: 0.774; p&lt;0.01), as well as between the body length and testes (rs: 0.604 and 0.659; p&lt; 0.05), the body length and the length of uterus (rs: 0.839; p&lt; 0,01) and between the ovary width and egg length (rs: 0.777; p&lt;0.01). Morphological study of the pre-ovigerous adults demonstrated that the ovary and testes develop simultaneously before the development of the uterus and vitelline glands. The acetabulum was detected in pre-ovigerous adults stained with hematoxilin and observed using light microscopy. In these specimens, the acetabulum measured 36.7 ± 6.9 µm (25-50 µm) in width and 39.91 ± 6.8 µm (25-55 µm) in length. The acetabulum was not detected in post-ovigerous adults observed with light microscopy. However, this structure was detected using confocal miscrocopy. In the post-ovigerous specimens, the acetabulum presented a reduced size compared to the pre-ovigerous adults. This may imply that this structure has more functional significance in the larval and pre-ovigerous stages.

Four whole-istic aspects of schistosome granuloma biology: fractal arrangement, internal regulation, autopoietic component and closure

Submitted by Gentil Jeorgina ([email protected]) on 2012-01-27T14:58:48Z No. of bitstreams: 1 Four whole-istic aspects of schistosome granuloma biology.pdf: 27136048 bytes, checksum: 68f5b106c7608be95432b04c157a4a4a (MD5)Made available in DSpace on 2012-01-27T14:58:48Z (GMT). No. of bitstreams: 1 Four whole-istic aspects of schistosome granuloma biology.pdf: 27136048 bytes, checksum: 68f5b106c7608be95432b04c157a4a4a (MD5) Previous issue date: 2006Financial support: CNPq, FiocruzFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Patologia. Rio de Janeiro, RJ, BrasilFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Patologia. Rio de Janeiro, RJ, BrasilUniversidade Federal de Pernambuco. Departamento de Física. Recife, PE, BrasilUniversidade Federal da Paraíba. Departamento de Física. João Pessoa, PB, BrasilFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Patologia. Rio de Janeiro, RJ, BrasilFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Patologia. Rio de Janeiro, RJ, BrasilFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Patologia. Rio de Janeiro, RJ, BrasilFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Patologia. Rio de Janeiro, RJ, BrasilThis paper centers on some whole-istic organizational and functional aspects of hepatic Schistosoma mansoni granuloma, which is an extremely complex system. First, it structurally develops a collagenic topology, originated bidirectionally from an inward and outward assembly of growth units. Inward growth appears to be originated from myofibroblasts derived from small portal vessel around intravascular entrapped eggs, while outward growth arises from hepatic stellate cells. The auto-assembly of the growth units defines the three-dimensional scaffold of the schistosome granulomas. The granuloma surface irregularity and its border presented fractal dimension equal to 1.58. Second, it is internally regulated by intricate networks of immuneneuroendocrine stimuli orchestrated by leptin and leptin receptors, substance P and Vasoactive intestinal peptide. Third, it can reach the population of ± 40,000 cells and presents an autopoietic component evidenced by internal proliferation (Ki-67+ Cells), and by expression of c-Kit+ Cells, leptin and leptin receptor (Ob-R), granulocyte-colony stimulating factor (G-CSF-R), and erythropoietin (Epo-R) receptors. Fourth, the granulomas cells are intimately connected by pan-cadherins, occludin and connexin-43, building a state of closing (granuloma closure). In conclusion, the granuloma is characterized by transitory stages in such a way that its organized structure emerges as a global property which is greater than the sum of actions of its individual cells and extracellular matrix components

Histological Analyses Demonstrate the Temporary Contribution of Yolk Sac, Liver, and Bone Marrow to Hematopoiesis during Chicken Development

<div><p>The use of avian animal models has contributed to the understanding of many aspects of the ontogeny of the hematopoietic system in vertebrates. However, specific events that occur in the model itself are still unclear. There is a lack of consensus, among previous studies, about which is the intermediate site responsible for expansion and differentiation of hematopoietic cells, and the liver's contribution to the development of this system. Here we aimed to evaluate the presence of hematopoiesis in the yolk sac and liver in chickens, from the stages of intra-aortic clusters in the aorta-genital ridges-mesonephros (AGM) region until hatching, and how it relates to the establishment of the bone marrow. <i>Gallus gallus domesticus</i> L. embryos and their respective yolk sacs at embryonic day 3 (E3) and up to E21 were collected and processed according to standard histological techniques for paraffin embedding. The slides were stained with hematoxylin-eosin, Lennert's Giemsa, and Sirius Red at pH 10.2, and investigated by light microscopy. This study demonstrated that the yolk sac was a unique hematopoietic site between E4 and E12. Hematopoiesis occurred in the yolk sac and bone marrow between E13 and E20. The liver showed granulocytic differentiation in the connective tissue of portal spaces at E15 and onwards. The yolk sac showed expansion of erythrocytic and granulocytic lineages from E6 to E19, and E7 to E20, respectively. The results suggest that the yolk sac is the major intermediate erythropoietic and granulopoietic site where expansion and differentiation occur during chicken development. The hepatic hematopoiesis is restricted to the portal spaces and represented by the granulocytic lineage.</p></div

The embryonic development of Schistosoma mansoni eggs: proposal for a new staging system

Submitted by Gentil Jeorgina ([email protected]) on 2012-07-18T17:41:30Z No. of bitstreams: 1 The embryonic development of Schistosoma mansoni.pdf: 748130 bytes, checksum: f033be092ab04d0c3300206d02ce5804 (MD5)Made available in DSpace on 2012-07-18T17:41:30Z (GMT). No. of bitstreams: 1 The embryonic development of Schistosoma mansoni.pdf: 748130 bytes, checksum: f033be092ab04d0c3300206d02ce5804 (MD5) Previous issue date: 2009Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Patologia. Rio de Janeiro, RJ, Brasil / Instituto Gulbenkian de Ciência. Oeiras, Portugal.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Patologia. Rio de Janeiro, RJ, Brasil / Instituto Gulbenkian de Ciência. Oeiras, Portugal.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Patologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Esquistossomose Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Patologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Patologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Esquistossomose Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Patologia. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio Janeiro. Instituto de Microbiologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Esquistossomose Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Patologia. Rio de Janeiro, RJ, Brasil.Schistosomiasis is a water-borne parasitic illness caused by neoophoran trematodes of the genus Schistosoma. Using classical histological techniques and whole-mount preparations, the present work describes the embryonic development of Schistosoma mansoni eggs in the murine host and compares it with eggs maintained under in vitro conditions. Two pre-embryonic stages occur inside the female worm: the prezygotic stage is characterized by the release of mature oocytes from the female ovary until its fertilization. The zygotic stage encompasses the migration of the zygote through the ootype, where the eggshell is formed, to the uterus. Fully formed eggs are laid still undeveloped, without having suffered any cleavage. In the outside environment, eight embryonic stages can be defined: stage 1 refers to early cleavages and the beginning of yolk fusion. Stage 2 represents late cleavage, with the formation of a stereoblastula and the onset of outer envelope differentiation. Stage 3 is defined by the elongation of the embryonic primordium and the onset of inner envelope formation. At stage 4, the first organ primordia arise. During stages 5 to 7, tissue and organ differentiation occurs (neural mass, epidermis, terebratorium, musculature, and miracidial glands). Stage 7 is characterized by the nuclear condensation of neurons of the central neural mass. Stage 8 refers to the fully formed larva, presenting muscular contraction, cilia, and flame-cell beating. This staging system was compared to a previous classification and could underlie further studies on egg histoproteomics (morphological localizome). The differentiation of embryonic structures and their probable roles in granulomatogenesis are discussed herein

Chicken liver development without hematopoietic activity from E9 to E 14.

<p>The embryonic day (E) is indicated in the upper right corner in each picture. Hp, hepatoblasts; CV, central vein; sinusoidal capillaries (asterisks); PV, portal vein; CT, connective tissue. (<b>A</b>) Lennert's Giemsa and (<b>B</b>–<b>F</b>) Hematoxylin-eosin stains. Bars 20 µm.</p

Chicken liver development without hematopoietic activity from E3 to E8.

<p>The embryonic day (E) is indicated in the upper right corner in each picture. (<b>A</b>, <b>C</b>, <b>D</b>) Numerous mitosis (arrows) are seen in hepatoblasts (Hp). (<b>B</b>) Mitosis in circulating erythrocyte (arrow). (<b>E</b>) Immature hematopoietic circulating cells (arrows) in sinusoidal capillaries (vessels located between hepatoblast cords, Hp). Note the large and irregular lumen of the sinusoidal capillaries. (<b>F</b>) Foci of immature erythropoietic cells in circulation (limited by arrows). Hp, hepatoblasts; VD, venous duct; sinusoidal capillaries (asterisks). (<b>A</b>, <b>C</b>, <b>F</b>) Hematoxylin-eosin and (<b>B</b>, <b>D</b>, <b>E</b>) Lennert's Giemsa stains. Bars 20 µm.</p

(A–K) Erythropoiesis (Ery) and (E–L) granulopoiesis (Gr) in chicken yolk sac between E3 and E20.

<p>The embryonic day (E) is indicated in the upper right corner in each picture. Thin arrows show mitosis in erythrocytes (<b>A</b>, <b>D</b>, <b>E</b>, <b>I</b>, <b>K</b>) and in a granulocyte (<b>H</b>). (<b>A</b>) Predominance of basophilic cells with a slight acidophily. (<b>B</b>, <b>C</b>) Numerous pro-erythroblasts and basophilic erythroblasts (Ery) between artery (Art) and endoderm (End). (<b>C</b>) Note a cell band leukocyte (arrowhead). (<b>D</b>) Erythrocytic (Ery) focus showing mature erythrocyte (arrowhead). (<b>F</b>, <b>G</b>) Cell band leukocyte (arrowhead). (<b>H</b>) Eosinophil granules into granulocytic cells at different stages of maturation show the cytoplasm of these cells in red-orange color. (<b>I</b>) Promyelocyte (arrowhead). (<b>J</b>, <b>K</b>) Foci of erythrocytic (white asterisks) and granulocytic (black asterisks) differentiation are present in equivalent numbers at this stage. (<b>K</b>) Mature leukocyte (arrowhead). (<b>L</b>) Myelocyte (arrowhead). (<b>E</b>–<b>K</b>) Note that granulocytic and erythrocytic lineages do not mix. End, endoderm; Art, artery. (<b>A</b>–<b>E</b>, <b>G</b>, <b>I</b>, <b>L</b>) Lennert's Giemsa, (<b>F</b>, <b>J</b>, <b>K</b>) Hematoxylin-eosin, and (<b>H</b>) Sirius Red stains at pH 10.2. Bars 20 µm.</p

Scheme of hematopoiesis in the yolk sac, liver, and bone marrow during chicken development.

<p>Bars indicate the temporal distribution of this activity in the AGM region, yolk sac (YS), liver (L), and bone marrow (BM). Black dotted lines indicate the presence of both erythropoiesis and granulopoiesis in the YS (<b>C</b>–<b>F</b>) and BM (<b>E</b>–<b>G</b>). The blue dotted line is to draw attention to the granulopoiesis in the L. (<b>A</b>) At E4, immature erythropoietic cells, incomplete erythropoietic foci, and rare granulocytes are distributed in the YS. (<b>B</b>) From E6, the YS shows complete erythropoietic foci (all maturation stages are seen) and some granulocytes. (<b>C</b>) From E7, complete erythropoietic and granulopoietic foci are distributed in the YS. (<b>D</b>) At E10, erythropoietic and granulopoietic foci are seen in the YS. Granulocytes at different stages of maturation are noted in the BM. At E11, basophilic cells are also seen in the BM. (<b>E</b>) At E15, both erythropoietic and granulopoietic foci are frequently observed in the YS and BM. In this phase, granulopoiesis begins in the L around the portal vessels. (<b>F</b>) At E17, hematopoiesis is reduced in the YS. Granulopoiesis persists in the L portal spaces, and both erythropoietic and granulopoietic activities are noted in the BM. (<b>G</b>) At E21, granulopoietic foci are seen in L connective tissues, both erythrocytic and granulocytic activities are observed in the BM, and the YS is no longer a hematopoietic site.</p