Intrauterine infection has been frequently linked with preterm labor before 30 wk of human pregnancy. Many different species of organisms have been detected, leading to the suggestion that infection-induced preterm labor is a generic inflammatory response to organisms rather than a specific response to a limited number of pathogens. The detection of organisms by microbiological culture is a laborious and unreliable process, so the aim of this study was to harness modern molecular techniques to detect organisms in tissues from human pregnancy. A DNA probe specific for conserved regions of bacterial 16S ribosomal RNA sequence was designed and labeled with fluorescein for fluorescence in situ hybridization. Organisms were detected in the great majority (Ͼ80%) of fetal membranes after prolonged premature rupture of the fetal membranes and after preterm labor, which was consistent with previous data. Organisms were also detected in fetal membranes after preterm delivery without labor and in term deliveries (with or without labour). Inflammatory cells were found frequently in the amnion or chorion of preterm fetal membranes but not in term tissues. Our primary finding is that fluorescence in situ hybridization is an appropriate method to detect organisms in human fetal membranes. In addition, our data show that bacteria may be present in fetal membranes without necessarily causing an inflammatory response, so the mere presence of bacteria may not be sufficient to cause preterm labor. Human labor at all gestational ages involves an inflammatory response, being characterized by increased levels of prostaglandins and cytokines (1,2). This inflammation is presumed to be initiated by physiological mediators, including corticotrophin-releasing hormone (3) or platelet-activating factor (4), or by pathological processes (5-7).At 23-32 wk of pregnancy, preterm labor is most frequently associated with micro-organisms within the uterus (8). These organisms are thought to activate inflammatory responses within intrauterine tissues and cause the recruitment of leukocytes to the fetal membranes (chorioamnionitis) (9,10). This is so thoroughly accepted that in some studies, chorioamnionitis has been used as being diagnostic of intrauterine infection, without determination of the presence of bacteria (11). However, the precise relation between the presence of bacteria and an inflammatory response has not been clearly defined. This is an important issue as it is not known whether the presence of bacteria always causes chorioamnionitis or whether chorioamnionitis is always linked to infection.Many different organisms have been identified from intrauterine tissues after preterm labor using various sampling and culture techniques (12-14), but no clear pattern has emerged from these studies, so it has not been possible to implicate one particular organism or family of organisms as the main causes of preterm labor. Furthermore, it has not been proved that the bacteria present in the vagina are those that have caused chorioamnionitis in pre...

A contaminant protein complex found in pharmaceutical urinary human chorionic gonadotrophin preparations is reported to have anti-human immunodeficiency virus-associated Kaposi's sarcoma activity. The aim of this study was to isolate and characterize this protein complex by proteomic approaches. Size exclusion chromatography was used in the isolation of these human chorionic gonadotrophin-associated fragments. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed the presence of a protein complex that dissociated into two protein bands under reducing conditions. Matrix-assisted laser desorption/ionization timeof-flight mass spectrometry of this complex showed three polypeptides at ϳ6.2, 11.4, and 15.8 kDa. Peptide mass mapping and N-terminal amino acid sequencing identified two polypeptides as metabolites of placental transforming growth factor-␤ (11.4 kDa) and bikunin (15.8 kDa). Subsequent matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis of the anti-human immunodeficiency virus-associated Kaposi's sarcoma active preparations CG-10 (Sigma), Pregnyl (Organon), and Profasi (Serono) revealed the presence of metabolites of placental transforming growth factor-␤ in all three; no other non-human chorionic gonadotrophin-related protein species were observed in these preparations. Our findings present evidence that urinary human chorionic gonadotrophin preparations are contaminated with metabolites of placental transforming growth factor-␤, which may have transforming growth factor-␤ agonist actions, and metabolites of bikunin, which is a protease inhibitor. In combination these molecules may be responsible for the anti-human immunodeficiency virus-associated Kaposi's sarcoma activity demonstrated for these urinary human chorionic gonad-