Maria Célia dos Santos

Effect of Macrophage Migration Inhibitory Factor (MIF) in Human Placental Explants Infected with Toxoplasma gondii Depends on Gestational Age

Because macrophage migration inhibitory factor (MIF) is a key cytokine in pregnancy and has a role in inflammatory response and pathogen defense, the objective of the present study was to investigate the effects of MIF in first- and third-trimester human placental explants infected with Toxoplasma gondii. Explants were treated with recombinant MIF, IL-12, interferon-γ, transforming growth factor-β1, or IL-10, followed by infection with T. gondii RH strain tachyzoites. Supernatants of cultured explants were assessed for MIF production. Explants were processed for morphologic analysis, immunohistochemistry, and real-time PCR analysis. Comparison of infected and stimulated explants versus noninfected control explants demonstrated a significant increase in MIF release in first-trimester but not third-trimester explants. Tissue parasitism was higher in third- than in first-trimester explants. Moreover, T. gondii DNA content was lower in first-trimester explants treated with MIF compared with untreated explants. However, in third-trimester explants, MIF stimulus decreased T. gondii DNA content only at the highest concentration of the cytokine. In addition, high expression of MIF receptor was observed in first-trimester placental explants, whereas MIF receptor expression was low in third-trimester explants. In conclusion, MIF was up-regulated and demonstrated to be important for control of T. gondii infection in first-trimester explants, whereas lack of MIF up-regulation in third-trimester placentas may be involved in higher susceptibility to infection at this gestational age.

Dopplervelocimetria colorida dos vasos orbitais: técnica de exame e anatomia vascular normal

Color dopplervelocimetry is an ultrasound modality that has rapidly expanded to all medical areas due to the large amount of information that can be obtained. Orbital color dopplervelocimetry has been used for the diagnosis of ophthalmic disorders and to study specific diseases such as preeclampsia. The aim of this study is to describe the normal anatomy of the orbit and the technical details of color dopplervelocimetry, since this is an accessible and reproducible technique. We believe that new applications for this technique will emerge and that it will be introduced in the daily practice of radiologists to improve the quality of diagnosis and follow-up of ocular and systemic diseases.

Doplervelocimetria das artérias oftálmica e central da retina em gestantes normais

OBJETIVOS: avaliar os padroes dos indices doplervelocimetricos das arterias oftalmica e central da retina durante o segundo e terceiro trimestres da gestacao normal e comparar os valores obtidos do olho direito e esquerdo das gestantes. METODOS: estudo transversal no qual se avaliaram seis indices doplervelocimetricos das arterias central da retina e oftalmica em 51 gestantes normais, com idades gestacionais entre a 20a e a 38a semana. As variaveis analisadas foram os indices de resistencia e pulsatilidade (IR, IP), os picos de velocidade sistolica e diastolica (PVS, PVD) e a razao entre picos de velocidade (RPV). A analise dos indices doplervelocimetricos dos olhos direito e esquerdo foi realizada utilizando-se a mediana dos valores. Para a comparacao dos valores dos indices entre os dois olhos das gestantes, utilizou-se o teste t de Student para dados pareados. A associacao entre a idade gestacional e os indices foi testada empregando-se o coeficiente de correlacao linear de Pearson. Adotou-se o nivel de significância de 5% para os testes estatisticos. RESULTADOS: a mediana dos indices doplervelocimetricos das arterias oftalmica e central da retina foram, respectivamente: IP=1,83; IR=0,78; PVS=34,20; PVD=6,80; RPV=0,48 e IP=1,34; IR=0,70; PVS=7,40; PVD=2,10. Nao houve diferencas na analise comparativa dos indices doplervelocimetricos entre os olhos direito e esquerdo das gestantes normais. O coeficiente de correlacao linear entre a idade gestacional e os indices de ambas as arterias nao mostrou diferenca significante durante a gestacao normal. CONCLUSAO: e factivel a analise unilateral dos indices doplervelocimetricos das arterias oftalmica e central da retina no estudo de doencas maternas sistemicas. Nao ha mudanca significativa dos indices doplervelocimetricos das arterias oftalmica e central da retina ao longo da gestacao normal entre a 20a e a 38a semana.

Padrão do consumo de álcool em gestantes atendidas em um hospital público universitário e fatores de risco associados

PURPOSE: To determine the pattern of alcohol use before and during pregnancy and associated risk factors in puerperal women hospitalized in a public university hospital in Southeastern Brazil. METHODS: Between June and September 2009, 493 puerperae were consecutively evaluated. Those with cognitive impairment were excluded from the study. The AUDIT and CAGE questionnaires were used to diagnose alcohol use/abuse before pregnancy, in addition to the T-ACE during pregnancy. Another questionnaire was applied to collect sociodemographic data, such as age, educational level, marital status, and household income. The χ2 test was used in the statistical analysis and the Odds Ratio (OR) and 95% confidence interval (95%CI) were calculated. A p-value <0.05 was considered to be significant. RESULTS: Before pregnancy, the CAGE was positive in 50/405 (12.3%) women and the AUDIT identified alcohol use in 331 (67.1%), which was of low risk in 233 (47.3%), risky in 73 (14.8%), and harmful or indicating possible alcohol dependence in 25 (5%). During pregnancy, the CAGE was positive in 53/405 (13.1%) women and the T-ACE in 84 (17%); the AUDIT identified alcohol use in 114 women, which was of low risk in 73 (14.8%), risky in 27 (5.5%), and harmful or indicating possible alcohol dependence in 14 (2.8%). During pregnancy, alcohol use was more frequent (OR=2.8; 95%CI 1.2 - 6.2) among women with a lower educational level (8.8 versus 3.3%) and more frequent (OR=3.8; 95%CI 1.3 - 11.1) among those who did not cohabit with a partner (6 versus 1.7%). Among pregnant women who drank alcohol, 49/114 (43%) were advised to stop drinking. CONCLUSIONS: Alarming alcohol use was observed during pregnancy, especially among pregnant women with a lower educational level and those who did not cohabit with a partner. There was a low frequency of counseling aimed at abstinence and the AUDIT was the instrument that most frequently diagnosed alcohol consumption.

Azithromycin is able to control Toxoplasma gondii infection in human villous explants

BackgroundAlthough Toxoplasma gondii infection is normally asymptomatic, severe cases of toxoplasmosis may occur in immunosuppressed patients or congenitally infected newborns. When a fetal infection is established, the recommended treatment is a combination of pyrimethamine, sulfadiazine and folinic acid (PSA). The aim of the present study was to evaluate the efficacy of azithromycin to control T. gondii infection in human villous explants.MethodsCultures of third trimester human villous explants were infected with T. gondii and simultaneously treated with either PSA or azithromycin. Proliferation of T. gondii, as well as production of cytokines and hormones by chorionic villous explants, was analyzed.ResultsTreatment with either azithromycin or PSA was able to control T. gondii infection in villous explants. After azithromycin or PSA treatment, TNF-α, IL-17A or TGF-β1 levels secreted by infected villous explants did not present significant differences. However, PSA-treated villous explants had decreased levels of IL-10 and increased IL-12 levels, while treatment with azithromycin increased production of IL-6. Additionally, T. gondii-infected villous explants increased secretion of estradiol, progesterone and HCG + β, while treatments with azithromycin or PSA reduced secretion of these hormones concurrently with decrease of parasite load.ConclusionsIn conclusion, these results suggest that azithromycin may be defined as an effective alternative drug to control T. gondii infection at the fetal-maternal interface.

Prospective study of the hemodynamic behavior of ophthalmic arteries in postpartum preeclamptic women: A doppler evaluation.

ABSTRACT The aim of this study was to evaluate the hemodynamic behavior of the ophthalmic artery by means of the Doppler ultrasound, in postpartum preeclamptic women. It was an observational prospective study with 44 postpartum preeclamptic women (group 1) and 49 postpartum normal women with normal blood pressure and with no previous illnesses known (group 2). All the pregnant women had a Doppler ultrasound exam of the ophthalmic artery in the immediate puerperium, that is, 10 days for the delivery (time 1). Group 1 was then followed prospectively, 26 patients of which returned to the last test in the remote puerperium in 45 days (time 2) and 29 patients returned to the last test in the remote puerperium in 90 days (time 3). All these women had preeclampsia before the delivery. The resistance index (RI), pulsatility index (PI), peak systolic velocity (PSV), end diastolic flow velocity (EDV), second peak of systolic velocity (P2), and the peak ratio (PR) were calculated. The data obtained are expressed in average and standard deviation, by using the Lilliefors test for normality. The average of the Doppler indexes in groups 1 and 2 was compared by means of test t of the student. Group 1 was analyzed separately, comparing the three times, using the test of ANOVA for repetitive measures and Tukey post-hoc range test. In the “Results” section, the statically meaningful differences in RI, PI, P2, RPV, and EDV (p < 0.0001, p < 0.0001, p < 0.0009, p < 0.0001, p < 0.0028) were found in the immediate puerperium of group 1 in relation to group 2, indicating the persistence of hyperperfusion and orbital vasodilatation in the immediate postpartum period in patients who had complicated pregnancies previously due to preeclampsia. In the evolutionary analysis of group 1, comparing the Doppler indexes between the immediate and late puerperium, statistically relevant differences between the rates of RI, P2, and PR (p < 0,01) were observed, showing a raise of RI and reduction of P2 and PR, and also a tendency of normalization of these rates in the late puerperium. When the same indexes were compared (PI, P2, and PR) now in times 2 and 3, the remote, and late puerperium, respectively, there were no significant differences, indicating the stabilization of these indexes since the 45th day of the puerperium. Within 90 days, RI, PI, and PR are not stabilized yet in relation to the control, even though there is a tendency of these indexes to reach the control. In conclusion, there was persistence of signs of vasodilatation and hyperperfusion of the orbital territory, represented by Doppler of the ophthalmic artery in the immediate puerperium of preeclamptic women. A tendency of normalization of the orbital hemodynamic standard in the pregnant women from the period of the late puerperium was observed, but there was no complete normalization of the vascular pattern on the remote postpartum.

Enrofloxacin and Toltrazuril Are Able to Reduce Toxoplasma gondii Growth in Human BeWo Trophoblastic Cells and Villous Explants from Human Third Trimester Pregnancy

Classical treatment for congenital toxoplasmosis is based on combination of sulfadiazine and pyrimethamine plus folinic acid. Due to teratogenic effects and bone marrow suppression caused by pyrimethamine, the establishment of new therapeutic strategies is indispensable to minimize the side effects and improve the control of infection. Previous studies demonstrated that enrofloxacin and toltrazuril reduced the incidence of Neospora caninum and Toxoplasma gondii infection. The aim of the present study was to evaluate the efficacy of enrofloxacin and toltrazuril in the control of T. gondii infection in human trophoblast cells (BeWo line) and in human villous explants from the third trimester. BeWo cells and villous were treated with several concentrations of enrofloxacin, toltrazuril, sulfadiazine, pyrimethamine, or combination of sulfadiazine+pyrimethamine, and the cellular or tissue viability was verified. Next, BeWo cells were infected by T. gondii (2F1 clone or the ME49 strain), whereas villous samples were only infected by the 2F1 clone. Then, infected cells and villous were treated with all antibiotics and the T. gondii intracellular proliferation as well as the cytokine production were analyzed. Finally, we evaluated the direct effect of enrofloxacin and toltrazuril in tachyzoites to verify possible changes in parasite structure. Enrofloxacin and toltrazuril did not decrease the viability of cells and villous in lower concentrations. Both drugs were able to significantly reduce the parasite intracellular proliferation in BeWo cells and villous explants when compared to untreated conditions. Regardless of the T. gondii strain, BeWo cells infected and treated with enrofloxacin or toltrazuril induced high levels of IL-6 and MIF. In villous explants, enrofloxacin induced high MIF production. Finally, the drugs increased the number of unviable parasites and triggered damage to tachyzoite structure. Taken together, it can be concluded that enrofloxacin and toltrazuril are able to control T. gondii infection in BeWo cells and villous explants, probably by a direct action on the host cells and parasites, which leads to modifications of cytokine release and tachyzoite structure.

76 Analysis of the ophthalmic artery doppler indexes in chronic arterial hypertension pregnant women

Objective To assess the Doppler indexes of the ophthalmic artery in pregnant women who have been diagnosed with chronic arterial hypertension, as well as to compare these data with a group of non-hypertension pregnant women and to identify the cut-off points for differentiating between the indexes of both groups. Method This investigation refers to transversal observational study, which assessed 220 pregnant women who have chronic arterial hypertension as well as non-hypertension pregnant women in their second and third trimesters constituting, respectively, the study group and the control group. All the patients underwent Doppler evaluation of the ophthalmic artery, with an evaluation of the resistance indexes (IR), pulsatility (IP), and ratio between velocity peaks (RPV). Results There was a meaningful difference between the averages of the Doppler indexes of the ophthalmic artery between the two groups assessed; showing lower IR and IP values and higher RPV in the study group in relation to the control group and it was possible to identify cut-off points for differentiation between the indexes of both groups Conclusion There are flow alterations in the ophthalmic arteries of pregnant women who have chronic arterial hypertension, represented by signs of moderate vasodilatation and impedance fall in the orbital territory, the fact which can be extrapolated for the small-caliber veins of the central nervous system, considering the ophthalmic arteries belong to the central nervous system. As to the discrimination between the groups, the ratio between velocity peaks has shown to be the best index

Estudo do perfil hemodinâmico da artéria oftálmica em gestantes com diabetes mellitus gestacional

PURPOSES To evaluate the hemodynamic patterns of the ophthalmic artery by Doppler analysis in women with gestational diabetes mellitus (GDM), comparing them to normal pregnant women. METHODS A prospective case-control study that analyzed the ophthalmic artery Doppler indices in two groups: one consisting of 40 women diagnosed with GDM and the other of 40 normal pregnant women. Included were pregnant women with GDM criteria of the American Diabetes Association-2012, with 27 weeks of pregnancy to term, and excluded were women with hypertension, use of vasoactive drugs on or previous diagnosis of diabetes. Doppler analysis was performed in one eye with a 10 MHz linear transducer and the Sonoace 8000 Live Medison® equipment. The following variables were analyzed: pulsatility index (PI), resistance index (RI), peak velocity ratio (PVR), peak systolic velocity (PSV) and end diastolic velocity (EDV). To analyze the normality of the samples we used the Lillefors test, and to compare means and medians we used the Students t-test and Mann-Whitney test according to data normality, with the level of significance set at 95%. RESULTS The median and mean values with standard deviation of the variables of the ophthalmic artery Doppler velocimetry group GDM and normal pregnant women were: IP=1.7±0.6 and 1.6±0.4 (p=0.7); IR=0.7 and 0.7 (p=0.9); RPV=0.5±0.1 and 0.5±0.1 (p=0.1), PSV=33.6 and 31.9 cm/sec (p=0.7); VDF=6.3 and 7.9 cm/sec (p=0.4). There was no significant difference in the means and medians of these variables between the two groups of pregnant women. CONCLUSIONS The ophthalmic artery hemodynamic patterns, analyzed by means of a Doppler technique remained unchanged in the group of pregnant women with GDM compared to the group of normal pregnant women, suggesting that the time of exposure to the disease during pregnancy was too short to cause significant vascular disorders in the central territory.