Receptores de estrógeno e progesterona em células do sedimento de fluido peritoneal na endometriose pélvica: estudo imunocitoquímico

Objetivo: avaliar a expressão de receptores de estrógeno (RE) e progesterona (RP) em esfregaços de sedimento de fluido peritoneal em pacientes com endometriose, comparando-a com a de mulheres sem endometriose. Métodos: foi realizado imunocitoquímica para RE e RP em esfregaços de sedimento de fluido peritoneal em 19 casos de endometriose e 7 sem endometriose (controle). Os dados foram submetidos ao teste t de Student para análise estatística. Resultados: em 84,6% dos casos de pacientes com endometriose, células tipo endometrial expressaram RE, numa média de 4,1%, ao passo que nos casos sem endometriose foi positivo em 42,9%, com uma expressão média de 4,5% (p = 0,1706). RP foi expresso apenas em um caso, com endometriose, com história de rotura de endometrioma. Conclusões: não houve diferença da expressão de RE entre os casos de endometriose e os casos-controle, observando-se um comportamento distinto em tecidos. Um mais aprofundado estudo deve ser feito para melhor avaliar o enigmático mecanismo envolvido na manifestação de receptores hormonais em células esfoliadas.

Concentrações de FSH, LH, estradiol, progesterona e histamina no soro, no fluido peritoneal e no fluido folicular de mulheres com e sem endometriose

OBJETIVOS: relatos da literatura mostram que não há dados conclusivos sobre a associação entre a endometriose e as concentrações de hormônios envolvidos no controle da reprodução. Este estudo foi realizado para determinar as concentrações de FSH, LH, estradiol (E), progesterona (P) e histamina (Hi) no soro, no fluido peritoneal e no fluido folicular de mulheres com e sem endometriose. MÉTODOS: a extensão da doença foi estadiada de acordo com a American Fertility Society classification (1997). Para a coleta de soro e de fluido peritoneal foram selecionadas 28 mulheres com endometriose submetidas à laparoscopia diagnóstica (18 mulheres inférteis com endometriose I-II e dez mulheres inférteis com endometriose III-IV). Para o grupo controle, foram selecionadas 21 mulheres férteis submetidas à laparoscopia para esterilização tubárea. O fluido folicular foi obtido de 39 mulheres inférteis submetidas a fertilização in vitro (21 mulheres com endometriose e 18 mulheres sem endometriose). RESULTADOS: as concentrações de FSH e LH no soro, no fluido peritoneal e no fluido folicular não diferiram significativamente entre os grupos. As concentrações de E e P no fluido peritoneal foram significativamente mais baixas em mulheres inférteis com endometriose (E: 154,2±15,3 para estágios I-II e 89,3±9,8 ng/mL para estágios III-IV; P: 11,2±1,5 para estágios I-II e 7,6±0,8 ng/mL para estágios III-IV) em comparação com mulheres controle (E: 289,1±30,1; P: 32,8±4,1 ng/mL) (Testes de Kruskal-Wallis/Dunn; p<0,05). No soro, as concentrações de E e P seguiram o mesmo padrão. No fluido folicular, as concentrações de E e Hi foram significativamente mais baixas em mulheres com endometriose (E: 97,4±11,1 ng/mL; Hi: 6,6±0,9 ng/mL) em comparação com mulheres sem endometriose (E: 237,5±28,5 pg/mL; Hi: 13,8±1,3 ng/mL), enquanto os níveis de P não revelaram diferença significativa entre os grupos (teste t de Student; p<0,05). CONCLUSÕES: nossos resultados indicam disfunção ovariana em mulheres com endometriose, com redução nas concentrações de estradiol, progesterona e histamina, o que pode contribuir para a subfertilidade freqüentemente associada à doença.

Passagem de células endometriais para a cavidade peritoneal durante histeroscopia diagnóstica

OBJETIVO: avaliar a passagem de células endometriais para a cavidade peritoneal durante histeroscopia diagnóstica. MÉTODOS: estudo descritivo, prospectivo, envolvendo 61 pacientes sem afecção endometrial maligna e 15 com câncer do endométrio. Duas amostras de lavado peritoneal foram colhidas, uma antes (LP-1) e outra (LP-2), imediatamente após a realização da histeroscopia diagnóstica. A passagem para a cavidade peritoneal foi definida como a presença de células endometriais no LP-2, devendo tais células estarem ausentes no LP-1. Utilizou-se histeroscópio com 5 mm de diâmetro (Storz). O meio de distensão foi o CO2 com pressão de fluxo de 80 mmHg controlada eletronicamente. O LP foi fixado em álcool absoluto (1:1). As lâminas foram preparadas pelo método de Papanicolaou e todas as leituras feitas pelo mesmo observador. RESULTADOS: foram excluídas quatro pacientes (5,26%) por apresentarem células endometriais no LP-1, sendo duas em cada grupo. Nas 72 restantes, não houve passagem de células para a cavidade peritoneal. No grupo sem afecção maligna endometrial, 88,1% (52/59) apresentaram endométrio secretor, com correlação de 80% entre o diagnóstico histeroscópico e a biópsia do endométrio. No grupo com afecção maligna endometrial, a maioria das pacientes encontrava-se no estádio I (92,3%). A correlação entre histeroscopia/biópsia endometrial e exame anatomopatológico da peça cirúrgica foi de 100%. CONCLUSÕES: a realização de histeroscopia diagnóstica com CO2 e pressão de fluxo de 80 mmHg não determinou passagem de células endometriais para a cavidade peritoneal em ambos os grupos, sugerindo que a histeroscopia é método seguro nas pacientes com suspeita de câncer endometrial.

Peritoneal fluid changes in horses subjected to small colon distension

Intestinal devitalization in cases of small colon obstruction may be difficult to detect based only in clinical signs. The purpose was to serially evaluate blood and peritoneal fluid of horses subjected to small colon distension. Seventeen adult horses were allotted in three groups. In the small colon-distended group (DG, n=7) a surgically-implanted latex balloon was inflated to promote intraluminal small colon distension. In the shamoperated group (SG, n=5), the balloon was implanted but not inflated, and no surgery was done in the control group (CG, n=5). Blood and peritoneal fluid were sampled before and after (6 samples with a 30-minute interval) intestinal obstruction for cytological and biochemical analyses. No significant changes in clinical signs occurred within groups or across time during the experimental period. There were no statistical differences among SG and SG groups in hematologic and blood chemistry variables. Although total protein concentration and lactate dehydrogenase (LDH) activity in peritoneal fluid remained most of the time within reference values during the experimental period in all groups, increases from baseline values were detected in SG and DG groups. Such increases occurred earlier, progressively and with greater magnitude in the DG when compared with the SG (P<0.05). Increases from baselines values were also observed in total nucleated cells and neutrophils counts in the DG (P<0.05). In conclusion, distension of the equine small colon induced progressive subtle increases in total protein and LDH concentrations in the peritoneal fluid during the first hours. Serial evaluation of these variables in peritoneal fluid may be useful for early detection of intestinal devitalization in clinical cases of equine small colon obstruction.

Inflammatory and anti-inflammatory effects of soybean agglutinin

Soybean agglutinin (SBA) lectin, a protein present in raw soybean meals, can bind to and be extensively endocytosed by intestinal epithelial cells, being nutritionally toxic for most animals. In the present study we show that SBA (5-200 µg/cavity) injected into different cavities of rats induced a typical inflammatory response characterized by dose-dependent exudation and neutrophil migration 4 h after injection. This effect was blocked by pretreatment with glucocorticoid (0.5 mg/kg) or by co-injection of N-acetyl-galactosamine (100 x [M] lectin), but not of other sugars (100 x [M] lectin), suggesting an inflammatory response related to the lectin activity. Neutrophil accumulation was not dependent on a direct effect of SBA on the macrophage population since the effect was not altered when the number of peritoneal cells was increased or decreased in vivo. On the other hand, SBA showed chemotactic activity for human neutrophils in vitro. A slight increase in mononuclear cells was observed 48 h after ip injection of SBA. Phenotypic analysis of these cells showed an increase in the CD4+/CD8- lymphocyte population that returned to control levels after 15 days, suggesting the development of an immune response. SBA-stimulated macrophages presented an increase in the expression of CD11/CD18 surface molecules and showed some characteristics of activated cells. After intravenous administration, SBA increased the number of circulating neutrophils and inhibited in a dose-dependent manner the neutrophil migration induced by ip injection of carrageenan into peritoneal cavities. The co-injection of N-acetyl-galactosamine or mannose, but not glucose or fucose, inhibited these effects. The data indicate that soybean lectin is able to induce a local inflammatory reaction but has an anti-inflammatory effect when present in circulating blood

Rotavirus and reovirus interaction with mouse peritoneal resident phagocytic cells

Rotaviruses and reoviruses are involved in human and animal diseases. It is known that both viruses penetrate the gastrointestinal tract but their interaction with phagocytic cells is unknown. To study this interaction, peritoneal resident phagocytic cells were used and rotavirus and reovirus replication in peritoneal phagocytic cells was observed. However, rotavirus replication in these cells led to the production of defective particles since MA-104 cells inoculated with rotavirus phagocytic cell lysate did not show any evidence of virus replication. On the basis of these results, we suggest that, although reovirus dissemination may be helped by these phagocytic cells, these cells may control rotavirus infection and probably contribute to the prevention of its dissemination.

Cytokines produced by susceptible and resistant mice in the course of Paracoccidioides brasiliensis infection

Paracoccidioidomycosis (PCM) is the most prevalent deep mycosis in Latin America and presents a wide spectrum of clinical manifestations. We established a genetically controlled murine model of PCM, where A/Sn mice develop an infection which mimics the benign disease (immune responses which favor cellular immunity) and B10.A animals present the progressive disseminated form of PCM (preferential activation of B cells and impairment of cellular immune responses). To understand the immunoregulatory phenomena associated with resistance and susceptibility in experimental PCM, A/Sn and B10.A mice were studied regarding antigen-elicited secretion of monokines (TNF-a and TGF-ß) and type-1 (IL-2 and IFN-g) and type-2 (IL-4,5,10) cytokines. Total lymph node cells from resistant mice infected ip with P. brasiliensis produced early and sustained levels of IFN-g and IL-2; type-2 cytokines (IL-4 and IL-5) started to appear 8 weeks after infection. In contrast, susceptible mice produced low levels of IFN-g concomitant with significant levels of IL-5 and IL-10 early in the infection. In the chronic phase of the disease, susceptible animals presented a transitory secretion of IL-2, and IL-4. In the pulmonary infection IL-4, IL-5 and IL-10 were preferentially detected in the lung cells washings of susceptible animals. After in vitro challenge with fungal antigens, normal peritoneal macrophages from B10.A mice secreted high levels of TGF-ß and low levels of TNF-a. In contrast, macrophages from A/Sn animals released high levels of TNF-a associated with a small production of TGF-ß. The in vivo depletion of IFN-g not only abrogated the resistance of A/Sn mice but also diminished the relative resistance of B10.A animals. The in vivo depletion of IL-4 did not alter the disease outcome, whereas administration of rIL-12 significantly enhanced resistance in susceptible animals. Taken together, these results suggest that an early secretion of high levels of TNF-a and IFN-g followed by a sustained secretion of IL-2 and IFN-g plays a dominant role in the resistance mechanisms to P. brasiliensis infection. In contrast, an early and ephemeral secretion of low levels of TNF-a and IFN-g associated with production of IL-5, IL-10 and TGF-ß characterizes the progressive disease of susceptible animals.

Thioglycollate-elicited murine macrophages are cytotoxic to Mycoplasma arginini-infected YAC-1 tumor cells

Macrophages are important components of natural immunity involved in inhibition of tumor growth and destruction of tumor cells. It is known that these cells can be activated for tumoricidal activity by lymphokines and bacterial products. We investigated whether YAC-1 tumor cells infected with Mycoplasma arginini stimulate nitric oxide (NO) release and macrophage cytotoxic activity. Thioglycollate-elicited macrophages from male BALB/c mice were co-cultured for 20 h with YAC-1 tumor cells infected or not with Mycoplasma arginini. The cytotoxic activity was evaluated by MTT assay and nitrite levels were determined with the Griess reagent. Thioglycollate-elicited macrophages co-cultured with noninfected YAC-1 cells showed low cytotoxic activity (34.7 ± 8.6%) and low production of NO (4.7 ± 3.1 µM NO2-). These macrophages co-cultured with mycoplasma-infected YAC-1 cells showed significantly higher cytotoxic activity (61.4 ± 9.1%; P<0.05) and higher NO production (48.5 ± 13 µM NO2-; P<0.05). Addition of L-NAME (10 mM), an inhibitor of NO synthesis, to these co-cultures reduced the cytotoxic activity to 37.4 ± 2% (P<0.05) and NO production to 3 ± 4 µM NO2- (P<0.05). The present data show that Mycoplasma arginini is able to induce macrophage cytotoxic activity and that this activity is partially mediated by NO.