José Antonio Picanço Diniz

Sulfated Polysaccharides from the Egg Jelly Layer Are Species-specific Inducers of Acrosomal Reaction in Sperms of Sea Urchins

We have characterized the fine structure of sulfated polysaccharides from the egg jelly layer of three species of sea urchins and tested the ability of these purified polysaccharides to induce the acrosome reaction in spermatozoa. The sea urchin Echinometra lucunter contains a homopolymer of 2-sulfated, 3-linked α-L-galactan. The species Arbacia lixula and Lytechinus variegatus contain linear sulfated α-L-fucans with regular tetrasaccharide repeating units. Each of these sulfated polysaccharides induces the acrosome reaction in conspecific but not in heterospecific spermatozoa. These results demonstrate that species specificity of fertilization in sea urchins depends in part on the fine structure of egg jelly sulfated polysaccharide.

Isolation of Fonsecaea pedrosoi from thorns of Mimosa pudica, a probable natural source of chromoblastomycosis

We report the isolation of Fonsecaea pedrosoi from thorns of the plant Mimosa pudica L. at the place of infection identified by one of our patients. Clinical diagnosis of chromoblastomycosis was established by direct microscopic examination and cultures from the patients lesion. The same species was isolated from the patient and from the plant. Scanning electron microscopy of the surface of the thorns showed the characteristic conidial arrangement of F. pedrosoi. These data indicate that M. pudica could be a natural source of infection for the fungus F. pedrosoi.

Development of Natural Culture Media for Rapid Induction of Fonsecaea pedrosoi Sclerotic Cells In Vitro

ABSTRACT Fonsecaea pedrosoi is the main agent of chromoblastomycosis, a skin disease presenting verrucous lesions, in which round, thick-walled sclerotic cells are found. In vitro induction of sclerotic cells is time-consuming (20 to 45 days) and temperature dependent. We present two new natural media that reduce the sclerotic-cell induction time to only 2 days.

Characterization of Minaçu virus (Reoviridae: Orbivirus) and pathological changes in experimentally infected newborn mice.

Minaçu virus was isolated from Ochlerotatus scapularis (Diptera: Culicidae) in Minaçu, Goiás State, Brazil, in 1996. In attempting characterization of virus serological (hemagluttination inhibition, HI; indirect immunofluorescence assay, IFA), physicochemical [test for deoxycholate acid (DCA) sensitivity; polyacrylamide gel electrophoresis (PAGE)] tests and ultrastructural studies were made. Virus was also assayed in suckling mice after intracerebral inoculation of 0.02 ml and in VERO and C6/36 cells with 0.1 ml of viral suspension containing 105 LD50/ml. Inoculated and control systems were observed daily. Every 24 h, one control and two inoculated animals were killed for tissue testing, including histopathological changes by haematoxylin and eosin (HE)‐stained sections, which were semi‐quantified. Research into viral antigen in the tissues of mice [central nervous system (CNS), liver, heart, lungs, spleen and kidneys] was carried out by the immunohistochemical technique using the peroxidase system. The virus only replicated in VERO cells, with antigen positive by IFA. Positive complement fixation tests were only obtained using antiserum of Minaçu virus. Minaçu virus is DCA resistant; haemagglutinating activity was negative. By electronic microscopy non‐enveloped virus particles were 75 nm in diameter. PAGE analysis showed Minaçu virus genome profile with 10 RNA segments. Infected, non‐killed animals died 7 days after inoculation. Tissue lesions were observed in all organs, except the lungs. Intense lesions were observed in the CNS and the heart, where neurone and cardiocyte necroses, respectively, were noted. The liver, spleen and kidneys had moderate tissue changes. Viral antigens were more abundant in the CNS and the heart, and absent in the lungs. In conclusion, Minaçu virus belongs to the family Reoviridae, genus Orbivirus.

Characterization of two new rhabdoviruses isolated from midges (Culicoides SPP) in the Brazilian Amazon: Proposed members of a new genus, Bracorhabdovirus

Summary.Itacaiunas and Curionopolis viruses were isolated from Culicoides midges in Parauapebas municipality, Pará state, Brazil, in 1984 and 1985, respectively. Itacaiunas virus infected newborn mice and mosquito cells (C6/36), but did not replicate in some mammalian cell lineages; while Curionopolis virus infected only mice. Neither virus showed a serological relationship with any of the 195 known arboviruses circulating in Brazil, nor against 38 other rhabdoviruses isolated worldwide. Both virus particles are bullet-shaped and similar in morphology to that observed for other members of the family Rhabdoviridae. Partial nucleotide sequencing of the N protein showed that those two viruses constitute a separate clade in the family Rhabdoviridae, which we propose to be a new genus, designated Bracorhabdovirus.

The fine structure of Garnia gonadati and its association with the host cell

Abstract Most of the studies on the fine structure of protozoa of the Apicomplexa group have been carried out with members of the Toxoplasma, Eimeria, and Plasmodium genera. In the present study we analyzed the fine structure of Garnia gonadoti parasitizing the red blood cells of the Amazonian reptile Gonatodes humeralis (Reptilia: Lacertilia). Transmission electron microscopy of thin sections showed that G. gonadoti presented all structures characteristic of the group, including the apicoplast. However, four special features were observed: (1) absence of the hemozoin (malarial) pigment; (2) a group of microtubules associated with the mitochondrion; (3) a vacuole containing electron-dense material, which resembled the acidocalcisome described in trypanosomatids; and (4) a special array of the host-cell endoplasmic reticulum around the parasitophorous vacuole.

TGF-β plasma levels in chromoblastomycosis patients during itraconazole treatment

BACKGROUND Chromoblastomycosis (CBM) is a dermal mycosis. The disease evolves to a chronic state, presenting a suppurative granulomatous dermatitis, combined with variable dermal fibrosis. Pathogenesis of the inflammation and tissue repair in CBM are poorly understood. AIM To quantify Transforming Growth Factor-beta (TGF-beta) plasma levels of CBM patients during itraconazole (ITZ) treatment. METHODS Blood plasma of 12 CBM patients was subjected to TGF-beta titration with ELISA at 0, 3, 6 and 12months of 200mg per day of ITZ therapy, and correlated with the clinical aspects. Plasma of 12 healthy individuals were used for control. RESULTS CBM patients present high plasma levels of TGF-beta (7.016+/-1988pg/ml), decreasing after 03months (4.625+/-645pg/ml) of ITZ treatment, which correlates with a rapid clinical improvement. However, after 6 (6.566+/-777pg/ml) and 12months (6.908+/-776) of treatment, TGF-beta levels increase to almost the same levels observed before treatment, which is related to a slow clinical improvement, fungal persistence on the lesion, and fibrotic scars. CONCLUSION TGF-beta plasma levels are high in CBM patients. Fungal destruction by ITZ correlates with TGF-beta downregulation, but tissue remodeling and fungal persistence probably raises its levels again, interfering with cellular immune responses.

Presence of a protrusion on the ventral disk of adhered trophozoites of Giardia lamblia.

Abstract About 30% of the trophozoites of Giardia lamblia, fixed while adherent to the substrate using fixative solutions designed to better preserve cytoskeletal elements, showed the presence of a ventral disk. This structure varied in shape and size and could be seen by scanning and transmission electron microscopy, as well as by confocal laser scanning microscopy of cells incubated with 3-3′-dihexyloxacarbocyanine iodide, which labels cisternae of the endoplasmic reticulum. It could also be observed in living adherent cells. A row of microtubules, a large number of glycogen particles, peripheral vesicles and concentric membranes were seen within the protrusion.

Some observations on the fine structure of trophozoites of the haemogregarine Cyrilia lignieresi (Adeleina: Haemogregarinidae) in erythrocytes of the fish Synbranchus marmoratus (Synbranchidae)

Abstract. This communication describes the fine structure of trophozoites of the haemogregarine Cyrilia lignieresi (Laveran, 1906) found in erythrocytes of the fresh-water fish Synbranchus marmoratus from Belém, Pará, North Brazil. The parasite possesses the usual structures, such as conoid, rhoptries and micronemes, seen in members of the phylum Apicomplexa. Three structures, however, appear to be characteristic features of this parasite. The parasitophorous vacuole is unusual in containing a large number of spherical bodies. Secondly, some of the dense bodies, which are usually spherical organelles, may appear as elongated structures. Thirdly, peculiar invaginations of the inner membrane appear to divide the parasite into compartments.

Three‐dimensional morphometric analysis of microglial changes in a mouse model of virus encephalitis: age and environmental influences

Many RNA virus CNS infections cause neurological disease. Because Piry virus has a limited human pathogenicity and exercise reduces activation of microglia in aged mice, possible influences of environment and aging on microglial morphology and behavior in mice sublethal encephalitis were investigated. Female albino Swiss mice were raised either in standard (S) or in enriched (EE) cages from age 2 to 6 months (young – Y), or from 2 to 16 months (aged – A). After behavioral tests, mice nostrils were instilled with Piry‐virus‐infected or with normal brain homogenates. Brain sections were immunolabeled for virus antigens or microglia at 8 days post‐infection (dpi), when behavioral changes became apparent, and at 20 and 40 dpi, after additional behavioral testing. Young infected mice from standard (SYPy) and enriched (EYPy) groups showed similar transient impairment in burrowing activity and olfactory discrimination, whereas aged infected mice from both environments (EAPy, SAPy) showed permanent reduction in both tasks. The beneficial effects of an enriched environment were smaller in aged than in young mice. Six‐hundred and forty microglial cells, 80 from each group were reconstructed. An unbiased, stereological sampling approach and multivariate statistical analysis were used to search for microglial morphological families. This procedure allowed distinguishing between microglial morphology of infected and control subjects. More severe virus‐associated microglial changes were observed in young than in aged mice, and EYPy seem to recover microglial homeostatic morphology earlier than SYPy . Because Piry‐virus encephalitis outcomes were more severe in aged mice, it is suggested that the reduced inflammatory response in those individuals may aggravate encephalitis outcomes.