Role of divalent cations, pH, cytoskeleton componentes and surface charge on the adhesion of Trichomonas vaginalis to a polystyrene substrate

The process of adhesion of three different strains of Trichomonas vaginalis to a polystyrene substrate was analysed. The process of adhesion was dependent on the time of incubation and the pH of the phosphate-buffered solution (PBS) in which the parasites were suspended. The highest indices of adhesion were observed after an incubation time of 60 min at pH 6.6. The adhesion index increased when the parasites were incubated in the presence of culture media or when Ca++ or Mg++ was added to the PBS solution, whereas cytochalasin B, trypsin or neuraminidase reduced adhesion. Incubation of the parasites in the presence of poly-L-lysine facilitated the process of adhesion. Incubation of the parasites or polystyrene beads in the presence of poly-L-lysine led to important changes in their surface charge.

Substâncias antifúngicas de Xylaria sp., um fungo endofítico isolado de Palicourea marcgravii (Rubiaceae)

Five compounds, 2-hexyl-3-methyl-butanodioic acid (1), cytochalasin D (2), 7-dechlorogriseofulvin (3), cytochalasin B (4) and griseofulvin (5), have been isolated from the endophytic fungus Xylaria sp., and their structures were elucidated on the basis of spectroscopic data. In the bioautography assay against Cladosporium cladosporioides and Cladosporium sphaerospermum, compounds 1 and 2 were found to be active while compounds 3, 4 and 5 did not show antifungal activity.

Mechanisms involved in calcium oxalate endocytosis by Madin-Darby canine kidney cells

Calcium oxalate (CaOx) crystals adhere to and are internalized by tubular renal cells and it seems that this interaction is related (positively or negatively) to the appearance of urinary calculi. The present study analyzes a series of mechanisms possibly involved in CaOx uptake by Madin-Darby canine kidney (MDCK) cells. CaOx crystals were added to MDCK cell cultures and endocytosis was evaluated by polarized light microscopy. This process was inhibited by an increase in intracellular calcium by means of ionomycin (100 nM; N = 6; 43.9% inhibition; P<0.001) or thapsigargin (1 µM; N = 6; 33.3% inhibition; P<0.005) administration, and via blockade of cytoskeleton assembly by the addition of colchicine (10 µM; N = 8; 46.1% inhibition; P<0.001) or cytochalasin B (10 µM; N = 8; 34.2% inhibition; P<0.001). Furthermore, CaOx uptake was reduced when the activity of protein kinase C was inhibited by staurosporine (10 nM; N = 6; 44% inhibition; P<0.01), or that of cyclo-oxygenase by indomethacin (3 µM; N = 12; 17.2% inhibition; P<0.05); however, the uptake was unaffected by modulation of potassium channel activity with glibenclamide (3 µM; N = 6), tetraethylammonium (1 mM; N = 6) or cromakalim (1 µM; N = 6). Taken together, these data indicate that the process of CaOx internalization by renal tubular cells is similar to the endocytosis reported for other systems. These findings may be relevant to cellular phenomena involved in early stages of the formation of renal stones.

Myosin V and iNOS expression is enhanced in J774 murine macrophages treated with IFN-gamma

Actin-based motor protein requirements and nitric oxide (NO) production are important features of macrophage activity during phagocytosis or microbicidal processes. Different classes of myosins contribute directly or indirectly to phagocytosis by providing mechanical force for phagosome closure or organelle movement. Recent data have shown the presence of myosins IC, II, V and IXb in phagosomes of bone marrow-derived murine macrophages. In our investigation we demonstrated the presence of different classes of myosins in J774 macrophages. We also analyzed the effect of gamma interferon (IFN-gamma), with or without calcium ionophore or cytochalasin B, on myosins as well as on inducible nitric oxide synthase (iNOS) expression and NO production. Myosins IC, II, Va, VI and IXb were identified in J774 macrophages. There was an increase of myosin V expression in IFN-gamma-treated cells. iNOS expression was increased by IFN-gamma treatment, while calcium ionophore and cytochalasin B had a negative influence on both myosin and iNOS expression, which was decreased. The increases in NO synthesis were reflected by increased iNOS expression. Macrophages activated by IFN-gamma released significant amounts of NO when compared to control groups. In contrast, NO production by calcium ionophore- and cytochalasin B-treated cells was similar to that of control cells. These results suggest that IFN-gamma is involved in macrophage activation by stimulating protein production to permit both phagocytosis and microbicidal activity.

Enucleated L929 mouse fibroblasts support invasion and multiplication of Shigella flexneri 5a

Invasive bacteria can induce their own uptake and specify their intracellular localization; hence it is commonly assumed that proximate modulation of host cell transcription is not required for infection. However, bacteria can also modulate, directly or indirectly, the transcription of many host cell genes, whose role in the infection may be difficult to determine by global gene expression. Is the host cell nucleus proximately required for intracellular infection and, if so, for which pathogens and at what stages of infection? Enucleated cells were previously infected with Toxoplasma gondii, Chlamydia psittaci, C. trachomatis, or Rickettsia prowazekii. We enucleated L929 mouse fibroblasts by centrifugation in the presence of cytochalasin B, and compared the infection with Shigella flexneri M90T 5a of nucleated and enucleated cells. Percent infection and bacterial loads were estimated with a gentamicin suppression assay in cultures fixed and stained at different times after infection. Enucleation reduced by about half the percent of infected cells, a finding that may reflect the reduced endocytic ability of L929 cytoplasts. However, average numbers of bacteria and frequency distributions of bacterial numbers per cell at different times were similar in enucleated and nucleated cells. Bacteria with actin-rich tails were detected in both cytoplasts and nucleated cells. Lastly, cytoplasts were similarly infected 2 and 24 h after enucleation, suggesting that short-lived mRNAs were not involved in the infection. Productive S. flexneri infection could thus take place in cells unable to modulate gene transcription, RNA processing, or nucleus-dependent signaling cascades.

Effects of the conditioned medium of mesenchymal stem cells on mouse oocyte activation and development

Mesenchymal stem cells (MSCs) have been reported to secrete a variety of cytokines and growth factors acting as trophic suppliers, but little is known regarding the effects of conditioned medium (CM) of MSCs isolated from femurs and tibias of mouse on the artificial activation of mouse oocytes and on the developmental competence of the parthenotes. In the current study, we investigated the effect of CM on the events of mouse oocyte activation, namely oscillations of cytosolic calcium concentration ([Ca²+]i), meiosis resumption, pronucleus formation, and parthenogenetic development. The surface markers of MSCs were identified with a fluorescence-activated cell sorter. The dynamic changes of the spindle and formation of pronuclei were examined by laser-scanning confocal microscopy. Exposure of cumulus-oocyte complexes to CM for 40 min was optimal for inducing oocyte parthenogenetic activation and evoking [Ca²+]i oscillations similar to those evoked by sperm (95 vs 100%; P > 0.05). Parthenogenetically activated oocytes immediately treated with 7.5 µg/mL cytochalasin B (CB), which inhibited spindle rotation and second polar body extrusion, were mostly diploid (93 vs 6%, P < 0.01) while CB-untreated oocytes were mostly haploid (5 vs 83%, P < 0.01). Consequently, the blastocyst rate was higher in the CB-treated than in the CB-untreated oocytes. There was no significant difference in developmental rate between oocytes activated with CM and 7% ethanol (62 vs 62%, P > 0.05), but the developmental competence of the fertilized oocytes was superior to that of the parthenotes (88 vs 62%, P < 0.05). The present results demonstrate that CM can effectively activate mouse oocytes, as judged by the generation of [Ca²+]i oscillations, completion of meiosis and parthenogenetic development.

Titulação de aloanticorpos anti-a e anti-b em gatos domésticos sem raça definida em Porto Alegre, Rio Grande do Sul, Brasil

A probabilidade de ocorrência de reação transfusional em um felino depende da prevalência local dos tipos sanguíneos felinos e dos títulos de aloanticorpos. A determinação dos títulos de aloanticorpos auxilia na estimativa do risco e da severidade de reação transfusional, após transfusão não compatível, em uma população de gatos. O objetivo deste trabalho foi determinar a titulação de aloanticorpos e o risco de possível reação transfusional, em felinos domésticos sem raça definida, da cidade de Porto Alegre. Foram selecionados aleatoriamente 100 gatos clinicamente saudáveis, sem raça definida e sem parentesco entre si e sem histórico de transfusão prévia. Amostras de sangue foram coletadas da veia jugular e a titulação de aloanticorpos foi determinada no soro de gatos com tipo sanguíneo previamente definido. O risco estimado foi calculado de acordo com estudos prévios. No presente trabalho, 82,5 e 100% dos gatos do tipo A e B, respectivamente, apresentaram titulação variada de aloanticorpos. Com base nos títulos encontrados, verificou-se que uma transfusão de sangue, do tipo A ou AB, em gatos do tipo B apresenta risco de 33,3% de reação aguda severa e de 66,7% de reação aguda leve, nos receptores felinos. A transfusão de sangue do tipo AB ou B em gatos do tipo A apresenta risco de reação aguda severa em 1,0%; reação aguda leve em 37,1% e destruição prematura dos eritrócitos em 44,3% dos receptores felinos.

DO ÍNDIO GENTIO AO GENTIO BÁRBARO: USOS E DESLIZES DA GUERRA JUSTA NA BAHIA SEISCENTISTA

Trata-se, neste artigo, de examinar o processo de legitimação que acompanhou a conquista do sertão baiano durante a segunda metade do século XVII, e de explorar as interações entre as dinâmicas locais e imperiais que levaram a uma situação de violência institucionalizada contra as populações indígenas do interior da Bahia.

Estudo sobre a cópula interespecífica entre moluscos planorbídeos Biomphalaria glabrata e B. tenagophila

Foi estudada a freqüência da cópula interespecífica entre espécimes de Biomphalaria glabrata e B. tenagophila. Os resultados da experiência levaram a conclusão de que há preferência pela cópula intraespecífica, ocorrendo, contudo, cópulas interespecíficas.

Evidenciação de antígeno da hepatite B (HBsAg) em "manchas" obtidas de exemplares de Triatominae: nota prévia

De 890 "manchas" provenientes de insetos dos gêneros Panstrongylus, Rhodnius e Triatoma, positivas para sangue humano, 7 deram resultados presumptivamente positivos por hemaglutinação passiva para HBsAg. Deste total só um caso foi confirmado como positivo, por RIE (radioimunoensaio), correspondendo a uma "mancha" obtida de Triatoma sordida (ninfa de 5.º estádio).

Evidenciação do antígeno da hepatite B (HBsAg) em Triatominae

De 3.200 "manchas" provenientes de insetos dos gêneros Triatoma e Panstrogylus, positivas para sangue humano, 12 deram resultados presuntivamente positivos, por imunodifusão, para HBsAg. Deste total, só 7 casos foram confirmados como positivos, por radioimunoensaio, correspondendo todos eles a "manchas" obtidas de ninfas de Triatoma infestans.

Condições higiênico-sanitárias do leite pasteurizado tipo "B" vendido na cidade de São Paulo, SP (Brasil), no período de fevereiro a agosto de 1982

Procedeu-se ao levantamento das condições higiênico-sanitárias de amostras de leite pasteurizado tipo "B", na cidade de São Paulo, SP (Brasil), nos meses de fevereiro a agosto de 1982. Trinta e cinco por cento das amostras analisadas apresentaram contagens excedentes a 10(4) psicrófilos/ml; 65%, excedentes a 4,0 x 10(4) mesófilos/ml; e, 63% superiores a 2 coliformes/ml. As presenças de Escherichia coli e de Staphylococcus aureus foram confirmadas em 17,5% das amostras analisadas, respectivamente.

Inibidores bacterianos no leite tipo ''B" comercializado no município de São Paulo, SP (Brasil)

No leite tipo "B", comercializado no município de São Paulo, SP (Brasil), foram pesquisadas a presença de inibidores bacterianos (penicilina, água oxigenada, formol e cloro). As amostras de leite utilizadas foram provenientes das quatro marcas de maior consumo pela população, coletadas no período de 14 de julho de 1982 a 20 de março de 1983. Foi constatada alta prevalência de inibidores bacterianos (4,70%). A incidência de resíduos de penicilina e de inibidores não identificados foi de 0,7% e de 3,40%, respectivamente. Houve uma baixa proporção de amostras com água oxigenada e formol e ausência de cloro.

Aflatoxina M1 no leite tipo "B" comercializado no Município de São Paulo, SP (Brasil)

No leite tipo "B", comercializado no Município de São Paulo, SP (Brasil), foi pesquisada a presença de aflatoxina M1. As amostras de leite analisadas foram provenientes das quatro marcas de maior consumo pela população, coletadas no período de julho a outubro de 1982. A aflatoxina M1, embora em baixos níveis e em pequena proporção (1,8%), fez-se presente nas quatro marcas.