Pseudomonas aeruginosa induces spatio-temporal secretion of IL-1ß, TNFα, proMMP-9, and reduction of epithelial E-cadherin in human alveolar epithelial type II (A549) cells.

Pseudomonas aeruginosa, is an opportunistic bacterium with a high prevalence in diverse pulmonary infections. Although several genes are involved in the system of resistance and evasion of the immunological response of the host, little is known about the inflammatory, degradative, and cell-binding response induced by P. aeruginosa in human lung alveolar epithelial cells. The purpose of this study was to determine the cytokine expression (IL-1ß and TNFα), pro matrix metalloproteinases activation (proMMP-2 and proMMP-9), and the effects on the cell-binding adhesion protein (E-cadherin) in an in vitro model of human lung alveolar epithelial cells. A549 cells were stimulated with a different number of colony-forming units of P. aeruginosa for 3, 6, and 24 hours. Subsequently, the culture medium was collected, IL-1ß and TNFα levels were evaluated by ELISA; proMMP-2 and -9 levels were determined by substrate gel zymography, and the MMP-9 and E-cadherin assessed by immunostaining of A549 cells. Our results demonstrated that P. aeruginosa induces mainly the secretion of TNFα, increases actMMP-9 level, and significantly reduces the level of E-cadherin in the A549 cells. In summary, the inflammatory/degradative process induced by P. aeruginosa modulates the expression of the E-cadherin protein. The probable clinical implications of this study suggest the use of inhibitors that reduce the degradative activity of proMMP-9 which will be further explored in the next phase of this study.

Eosinophilia in Preterm Born Infants Infected with Chlamydia trachomatis.

A higher than 350 eosinophils/mm(3) is strongly associated with Chlamydia trachomatis in term born babies coursing with respiratory distress. However, in preterm newborns infected with this pathogen, the levels of eosinophils are unknown. Forty newborn infants with clinical data of respiratory problems and/or sepsis were analyzed. DNA of leukocytes from peripheral blood was used to identify C. trachomatis. Detection of chlamydial infection was performed by amplifying the ompA gene by an in-house PCR, and eosinophil levels were evaluated in an XT-2000-hematology analyzer. Eighteen infants showed chlamydial infection and 14 of them showed pneumonia (RR = 2.6; CI95% 1.03-6.5; p =.027). Their eosinophil levels were 719 ± 614 cells/mm(3). A significant association between eosinophilia ≥1250 cells/mm(3) and gestational age of less than 29 weeks (RR = 5.8; 1.35; CI95% [1.4-24.5], p <.008) was observed. The preterm infants with chlamydial infection did not show higher eosinophil levels than uninfected infants.

Tissue-specific IL-10 secretion profile from term human fetal membranes stimulated with pathogenic microorganisms associated with preterm labor in a two-compartment tissue culture system.

OBJECTIVE: Interleukin (IL)-10 is a cytokine with anti-inflammatory properties that plays pivotal roles in immune recognition and maintenance of pregnancy, limiting the harmful effects of pro-inflammatory modulators. The aim of this work was to characterize the contribution of amnion and choriodecidua regions of the human fetal membranes in the production of IL-10 after selective stimulation with Candida albicans, Gardnerella vaginalis and Streptococcus agalactiae. METHODS: Pre-labor human fetal membranes were cultured in a two-compartment tissue culture system and stimulated with 1 × 10(6) CFU/ml of each pathogen added to either the amniotic or choriodecidual region or both. RESULTS: Candida albicans and G. vaginalis were the pathogens most effective in inducing IL-10 secretion, increasing 20 and 10 times, respectively, the levels of this cytokine in the choriodecidual compartment. Stimulation with S. agalactiae was effective only in the choriodecidual region, increasing two times IL-10 concentration. CONCLUSIONS: Synthesis and secretion of IL-10 in response to three different pathogens associated with intrauterine infection and preterm birth are differential and depend on the nature of the microorganism and initial contact region.

Progesterone elicits an inhibitory effect upon LPS-induced innate immune response in pre-labor human amniotic epithelium.

PROBLEM: Infection of human fetal membranes elicits secretion of pro-inflammatory modulators through its innate immune capacities. We investigated the effect of lipopolysacharide (LPS) and progesterone (P4) upon expression of TLR-4/MyD88, TNFα, IL-6, IL-8, IL-10, and HBD2 on the human amniotic epithelium. METHOD OF STUDY: Explants of the human amniotic epithelium were pre-treated with 0.01, 0.1, and 1.0 µM of P4; then cotreated with 1000 ng/mL LPS. TLR-4 was immuno-detected, and concentrations of MyD88, TNFα, IL-6, IL-8, IL-10, and HBD2 were quantified by ELISA. RESULTS: P4 significantly reduced the expression of LPS-induced TLR-4/MyD88. LPS increased the concentrations of TNFα, IL-6, IL-8, IL-10, and HBD2 by factors of 30-, eight, three, three, and fivefold, respectively. P4 at 1.0 µM was the most effective dose to blunt the secretion of TNFα, IL-6, and HBD-2. RU-486 blocks the effect of P4. CONCLUSION: P4 inhibited LPS-induced TLR-4/MyD88 and pro-inflammatory factors in the human amniotic epithelium. These results could explain partially how P4 can protect the amniotic region of fetal membranes and generate a compensatory mechanism that limits the secretion of pro-inflammatory modulators, which could jeopardize the immune privilege during pregnancy.

[Clinical efficacy of fluconazole, tinidazole and clindamycin vs fluconazole, tinidazole and azithromycin in the treatment of mixed cervical-vaginal infections, included those caused by Mycoplasma and Chlamydia trachomatis]. / Eficacia clínica de fluconazol, tinidazol y clindamicina vs fluconazol, tinidazol y azitromicina en el tratamiento de infecciones cérvico- vaginales mixtas, incluidas las causadas por Mycoplasma y Chlamydia trachomatis.

BACKGROUND: In the United States 19 million people acquire a sexually transmitted disease every year. Sexually transmitted diseases impact in gynecological terms because they may cause sterility, infertility and ectopic pregnancy. OBJECTIVE: To compare the effectiveness of two combinations of three oral antimicrobial drugs in the treatment of mixed cervical-vaginal infections, included those caused by Mycoplasma and Chlamydia trachomatis. MATERIAL AND METHOD: Aclinical, random, comparative, double-blind study included 50 patients assisting to infectology consult with diagnosis of mixed cervical-vaginal infection. Patients were divided into two groups: Group A (n = 25): fluconazole 37.5 mg, tinidazole 500 mg and azithromycin 250 mg; group B (n = 25): fluconazole 37.5 mg, tinidazole 500 mg and clindamycin 312.5 mg. Patients of both groups received two tablets twice p.o. for one day. Cultures were performed to corroborate the diagnosis and then to demonstrate effectiveness of the schemes studied. For the analysis of the data we used measures of central tendency, dispersion and inferential statistics for comparison of proportions by c2 and Fisher's exact tests with a significance level of p < 0.05. RESULTS: All patient got clinical cure; however, regarding the microbiologic eradication a positive case was identified in group A, requiring rescue treatment. The compliance in both groups was of 100%. In both groups, statistical analysis did not show significant differences. Three patients in group A had mild adverse effects. Patients mean age was 33.4 +/- 5.3 years. CONCLUSIONS: Both treatments showed similar effectiveness against mixed cervical-vaginal infections. Microbiological efficacy was of 96% and 100% in group A and B, respectively, besides, scheme of group B was better tolerated.

Tissue-specific human beta-defensins (HBD)-1, HBD-2 and HBD-3 secretion profile from human amniochorionic membranes stimulated with Candida albicans in a two-compartment tissue culture system.

BACKGROUND: During intrauterine infection, amniochorionic membranes represent a mechanical and immunological barrier against dissemination of infection. Human beta defensins (HBD)-1, HBD-2, and HBD-3 are key elements of innate immunity that represent the first line of defense against different pathogen microorganisms associated with preterm labor. The aim of this work was to characterize the individual contribution of the amnion (AMN) and choriodecidua (CHD) regions to the secretion of HBD-1, HBD-2 and HBD-3, after stimulation with Candida albicans. METHODS: Full-thickness human amniochorionic membranes were obtained after delivery by elective cesarean section from women at 37-40 wk of gestation with no evidence of active labor. The membranes were cultured in a two-compartment experimental model in which the upper compartment is delimited by the amnion and the lower chamber by the choriodecidual membrane. One million of Candida albicans were added to either the AMN or the CHD face or to both and compartmentalized secretion profiles of HBD-1, HBD-2, and HBD-3 were quantified by ELISA. Tissue immunolocalization was performed to detect the presence of HBD-1, -2, -3 in tissue sections stimulated with Candida albicans. RESULTS: HBD-1 secretion level by the CHD compartment increased 2.6 times (27.30 [20.9-38.25] pg/micrograms protein) when the stimulus with Candida albicans was applied only on this side of the membrane and 2.4 times (26.55 [19.4-42.5] pg/micrograms protein) when applied to both compartments simultaneously. HBD-1 in the amniotic compartment remained without significant changes. HBD-2 secretion level increased significantly in the CHD when the stimulus was applied only to this region (2.49 [1.49-2.95] pg/micrograms protein) and simultaneously to both compartments (2.14 [1.67- 2.91] pg/micrograms protein). When the stimulus was done in the amniotic compartment HBD-2 remained without significant changes in both compartments. HBD-3 remained without significant changes in both compartments regardless of the stimulation modality. Localization of immune-reactive forms of HBD-1, HBD-2, and HBD-3 was carried out by immunohistochemistry confirming the cellular origin of these peptides. CONCLUSION: Selective stimulation of amniochorionic membranes with Candida albicans resulted in tissue-specific secretion of HBD-1 and HBD-2, mainly in the CHD, which is the first region to become infected during an ascending infection.

Tumor necrosis factor in peritoneal fluid from asymptomatic infertile women

Background. Tumor necrosis factor-Ó (TNF-Ó) is a cytokine that can be found in the peritoneal fluid (PF) of patients with endometriosis and pelvic inflammatory disease (PID) as a response to inflammatory disorders and infections. The cytotoxic effect of this cytokine could be a factor participating in the pathology of various gynecological diseases, and could also be accountable for the high immunological response and demage to the tubal epithelium. The objective of this study was ato establish the presence of TNF-Ó in asymptomatic infertility and its association with various isolated bacteria. Methods. Ten milliliters of PF were collected from each of 73 patients by means of laparoscopy and cultured in synthetic medium and McCoy cells for the isolation of aerobic and anaerobic bacteria, as well as for Chlamydia trachomatis. The activity of TNF-Ó was determined by means of a bioassay using L-929 cells. Results. Forty-three parcent of the PFs showed positive TNF-Ó activity, while the laparoscopic evaluation showed that 32 patients had Fallopian tube occlusion (FTO), 7 had endometriosis, 30 had PID. and 4 had myomas and adhesions. TNF-Ó activity was found to be high in FTO patients (P<0.05). Positive cultures were found in 50.7 percent of patients; of these, 31.5 percent had PID (p< 0.05), and only 20.5 percent of positive cultures were TNF-Ó positive. Chlamydia trachomatis (16 percent) was the most frequently isolated bacteria in these patients. Conclusions. The detection of TNF-Ó could be useful in the diagnosis of active infectious and inflammatory diseases in asymptomatic infertile patients

Evaluación de la sensibilidad y especificidad de tres reactivos de inmunofluorescencia directa para el diagnóstico de Chlamydia trachomatis / Evaluation of sensitivity and specificity of three reagents of direct immunofluorescence for the diagnosis of chlamydia trachomatis

Se evaluó la sebsibilidad, especificidad, valor predictivo positivo y negativo y porcentaje de falsas positivas y negativas de tres reactivos de inmunofluorescencia directa para el diagnóstico de infecciones por Chlamydia trachomatis. Se estudiaron 150 muestras de raspados cervicovaginales y 50 muestras de líquido peritoneal de pacientes con enfermedad pélvica inflamatoria. Los reactivos en contra de la proteína principal de membrana externa presentaron mayor sensibilidad pero menor especifidad que el reactivo de inmunofluorescencia contra el lipopolisacárido de Chlamydia trachomatis

Infecciones ginecoobstétricas por bacterias anaerobias / Gyneco-obstetrical infections by anaerobic bactria

Entre los microorganismos (mo) de la flora normal del tracto genital femenino, las bacterias anaerobias (BA) predominando fue conocer la frecuencia de aislamiento y las características clínicas de las infecciones por BA en ginecoobstetricia; para lo cual se realizó un estudio retrospectivo mediante la revisión de los expedientes clínicos del total de pacientes GO que desarrollaron infecciones de las cuales se recuperaron BA, y que fueron manejadas por el servicio de infectología del Instituto Nacional de Perinatología (INPer), en el periódo comprendido del 1o de enero de 1988 a 31 de mayo de 1991. Se aislaron BA en 117 pacientes, las que desarrollaron 163 infecciones recuperándose de ellas 167 BA y 83 aerobias. El 98.2 por ciento fueron pacientes obstétricas. El 85.5 por ciento de los aislamientos de BA se obtuvieron de endometritis (EN), de abscesos de herida quirúrgica (HQ) se obtuvo 8.5 por ciento. La mayoría de las infecciones por anaerobios fueron polimicrobianas con una medida de 2.1 mo por infeción, en 58.1 por ciento de ellas también hubo aislamiento de bacterias aerobias. Las BA más frecuentes identificadas fueron los géneros Peptostreptococcus, Clostridium y bacteroides; entre los mo aerobios predominaron los Staphylococcus y las enterobacterias. La mayoria de las pacientes tuvieron una evolución clínica satisfactoria, controlándose la infección dentro de los primeros 5 días de tratamiento antimicrobiano. Durante el periódo estudiado no hubo defunciones. Concluimos que las BA tienen una participación importante en la etiología de las infecciones GO, siendo necesaria su búsqueda dirigida, y que el tratamiento incluya antibióticos contra estas bacterias.

Aspectos clínico-microbiológicos del Ureplasma urealyticum / Clinic and microbiological aspects of Ureoplasma urealyticum

Los micoplasmas revisten gran importancia clínica, por haberse reconocido su papel patógeno en entidades clínicas como neumonía atípica primaria, uretritis no gonocócica e infecciones cervicovaginales de la mujer en etapa sexual activa. En Europa durante el siglo XVIII, se presentó en el ganado una enfermedad pulmonar altamente contagiosa, a la cual se denominó pleuroneumonía. Sin embargo, fue hasta 1898 cuando Nocard y Roux lograron aislar el agente causal de esta enfermedad sin saber que se trataba de Mycoplasma mycoides. Las colonias poducidas en el medio de cultivo, se apreciaban muy pequeñas, pleomórficas y se teñían débilmente con los colorantes habituales. Con el paso del tiempo se siguieron observando algunos gérmenes, cuya morfología y propiedades de cultivo eran semejantes. Las fuentes de estos aislamientos fueron muy diversas. Al principio y de manera abreviada se les denominó PPLO (Pleuropneumoniae like organisms) por su similitud con agentes productores de pleuroneumonía bovina