417 The role of amniotic fluid nitric oxide and apoptosis in intra-amniotic infection

$194 SMFM Abstracts 415

416

MATRIX METALLOPROTEINASES IN CERVIC.M~VAGINAL FLUIDS AND THEIR CORRELATION WITH BACTERIAL PRESENCE AND PREGNANCY OUTCOME GIULIANA COATAL L PENNACCHIt, A RINELLP, L LIOTTAI, V LAURO 1, GIAN CARLO DIRENZOL ]Centre of Perinatal and Reproductive Medicine, Perugia OBJECTIVE: There is evidence that genital tract inflammation and infection may predispose to pretenn premature rapture of membranes (PPROM)and preterm labor. The aim of this study was to evaluate the relationship between presence of cervical-vaginal bacteria and concentration of matrix metalloproteinases (MMPs)-2 and -9 and their tissue inhibitor (TIMP-1) in the cervical-vaginal fluids and pregnancy outcome, STUDY DESIGN: CelMcal-vaginal fluids fi'om 122 consecutive pregnant women were obtained between 27-32 weeks of gestation. Microbiological swabs from vagina, cervix and urethr were taken for aerobic, anaerobic,and other c o m m o n microorganisms. MMP-9 and MMP-2 and TIMP-1 were measured by imnlunoassay. RESULTS: The rate of preterm delivery was 11.4% of which 96% had PPROM. Sixty-one percent of the patients that delivered at term and 71% of those delivered p r e t e r m were found to be positive for bacterial microorganisms at cexwical-vaginal level. MMP-9 cervical-vaginal concentrations were higher in the group of women that delivered at term than in the PPROM group (P = .02), while MMP-2 concentrations were higher in PPROM group than in the term delivery group (P - .05). The TIMP-1 concentration did not differ between the two groups. There was no significant correlation was found between bacterial presence and MMPs concentrations in both groups, while a significant correlation was observed between MMP-2 concentration and pregnancy outcome. CONCLUSION: This study demonstrated that an elevated MMP-2 concentration in mid-trimester cervical-vaginal fluids is associated with an increased risk of preterm labor, independent from the presence of cervicalvaginal infection. Considering that MMPs plays a central role in the basementm e m b r a n e degradation, we hypothesise that MMP-2 nfight up-regulate enzynmtic activity and therefore contribute to the oecurence of PPROM. Thus MMP-2 concentration assessment might play"a role in the prognosis of preterm labon

AMNIOTIC FLUID CHEMOKINE, LEUKOCYTE ADHESION MOLECULE, AND M A R I X METALLOPROTEINASE ARE INVOLVED IN INTRA-AMNIOTIC INFECTION CHAUR-DONG HSU 1, JIANN-HWA WANG1, KIRSTEN AVERSA 2, HASSAN HARIRAH3; lUniversity of Nebraska Medical Center, Obstetrics or Gynecology, Omaha, NE; 2yale University, Obstetrics or Gynecology, New Haven, CT; ~University of Texas Medical Branch at Galveston, Obstetrics and Gynecology, Galveston, TX OBJECTIVE: To investigate whether amrdotic fluid (AF) chemokine [interleukin-8 (IL-8)], leukocyte adhesion molecule-1 (L~M-1), and matrix metalloprotein-9 (MMP-9) were involved in the pathogenesis of intra-amnlotic injection (IAI). STUDY DESIGN: Sixty-six singleton pregnant women were studied. LM was defined as the presence of a positive AF culture. AE was tested for Gram stain, glucose, leukocytes, 1L-8, LAM-I, and MMP-9. AF IL-8, LAM-1, and MMP9 were d e t e r m i n e d by enzyme immunoassay. Statistical analyses were pet-formed using two-tailed t test, linear regression and correlation tests. Data were expressed as mean + SEM. RESULTS: Twenty-two patients were with IAI and 44 patients were not. There were no significant differences in maternal age, gestational age, parity, and race between patients with and without IAI. AF mean levels of IL-8. LAM1, and MMP-9 were significantly higher in pregnant women with IAI than in those without IAI (IL-8:131.91 _+28.13 vs 3.81 _+0.97 n g / m l , P< .0001; MMP-9: 231.13 _+46.53 vs 0.66 -+ 0.34 n g / m l , P < .0001; LAM-I: 82.27 + 12.51 vs 11.18 + 1.00 n g / m l , P< .0001). AF levels of tL-8, LAM-1, and MMP-9 were significantly correlated (IL-8/LAM-I: r = 0.61, P < .0001; IL-8/MMP-9: r = 0.80, P < .0001; IAM-1/MMP-9: r 0.80, P < .0001). Patients with 1AI had significantly higher AF leukocytes and significantly lower glucose levels than those without IAI. AF IL-8, LAM-1, and MMP-9 levels were significantly correlated with AF leukocytes (r = 0.68, P< .0001, and r 0.77, P< .0001, r - 0.82, P< .0001 respectively) and negatively correlated with AF glucose levels (r -0.49, P < .0001, r : -0.57, P < .000, and r = 0.52, P< .0001 respectively). CONCLUSION: Our data suggest AF IL-8 acts as an important chemoattractant and activator for the elevation ofAF LAM-1 and MMP-9 in patients with IAI. It also indicates that chemokine, adhesion molecule and matrix metalloproteinase may play an important role in the pathogenesis of IAI.

December 2001 A m J Obstet Gynecol 417

THE ROLE OF AMNIOTIC FLUID NITRIC OXIDE AND APOPTOSIS IN 1NTRA-AMNIOTIC INFECTION CHAUR-DONG HSU 1, JIANN-HWAWANG1, KIPSTEN AVERSA2, HASSAN HARIRAH3; tUniversity of Nebraska, Obstetrics or Gynecology, Omaha, NE; '-)Yale University, Obstetrics oi- Gynecology, New Haven, CT; 3University of Texas Medical Branch at Galveston, Obstetrics and Gynecology, Galveston, TX OBJECTIVE: To investigate the association and possible role of anmiotic fluid (AF) nitric oxide and apoptosis (nucleosome) in the pathogenesis of intra-amniotic infection (IAI), STUDY DESIGN: Sixty-one singleton p r e g n a n t w o m e n were studied. Twenty patients were with IAI and 41 patients were not. lAl was defined as the presence of a positive AF culture. AF was tested for Gram stain, glucose, leukocytes, nitric oxide (NOx: nitrate + nitrite), and nucleosome (Nu; end products of DNA fragmentation during apoptosis). Endogenous nitrite was determined using Greiss reagent. AF NOx was measured after the reduction of nitrate to nitrite with Aspergillus nitrate reductase. AF Nu was deternfined by enzyme immunoassay. AF NOx and Nu were normalized by AF creatinine levels. Statistical analyses were performed using two-tailed t test, linear regression and correlation tests. Data were expressed as mean + SEM. RESULTS: There were no significant differences in maternal age. gestational age, parity, and race between patients with and without IAI. AF mean levels of NOx, and Nu were significantly higher in patients with IAI than in those without IAI (NOx: 2.67 _+0.36 vs. 1.61 + 0.09 m m o l / m g creatinine, P < .001 and Nu: 90.07 + 28.17 vs. 0.00 _+0.00 U/nag creatinine, P< .0001). AF NOx levels were significantly correlated with Nu (r - 0.46, P < .001). Patients with IAI also had significantly higher AF leukocyte counts and significantly lower glucose levels than those without IAI. Both AF NOx and Nu levels were significantly correlated with AF leukocyte counts (r = 0.57, P < .0001, and r 0.60, P< .0001 respectively) and negatively correlated with AF glucose levels (r = -0.39, P : .002, and r = -0.40, P= .002 respectively'). CONCLUSION: Our data indicate that AF nitric oxide is associated with apoptosis and may play an important role in the pathogenesis of IAI. AF leukocytes are possibly one of the sources of elevated AF NOx and Nu in IAI.

418

DIAGNOSIS OF BV BY RAPID TESTING WITH A PH/AMINES TEST CARD J TOLOSA 1, S DALY2, T CHIPATO 3, C WHITNEY4, H GAITAN5, J SALWARIN6, P LUMBIGANON7, FOR THE GLOBAL NETWORK FOR PERINATAL AND REPRODUCTIVE HEALTH1; 1Thomas Jefferson University, Obstetrics and Gynecology, Philadelphia, PA; 2Coombe Women's Hospital, Dublin, Ireland; 3University of Zimbabwe, Harare; 4Centers for Disease Control and Prevention, Atlanta, GA; 5Universidad Nacional de Colombia, Saute Fe de Bogota; 6Population Council, Bangkok, Thailand 7Khon Kaen University, Khon Kaen, Thailand OBJECTIVE: To determine the ability of a new commercially-available rapid test (FemExam® pH and Amines Test Card TM, Litmus Concepts, Inc., Santa. Clara, CA) tot diagnosis of bacterial vaginosis (BV) compared to Gram stain. STUDY DESIGN: During 1999 to 2001, we collected lower vaginal swabs for Gram stain and rapid testing from asymptomatic women between 20 and 32 weeks gestation at centers in Colombia, Zimbabwe, Ireland, and the U.S. Study personnel p e r f o r m e d and read the rapid test iminediately following swab collection. The FemExam® has separate results windows for p H (positive if >4.7) and amines (positive if >0.5 mmol). According to the manufacturer, the test is positive when both are positive. Gram stains were performed and read at the University of Alabama, Birmingham. We used a Nugent's score of >7 to define BV. Sensitivity, specificity, positive and negative predictive values were calculated using Grain stain as the standard. RESULTS: Test results were available for 620 women; 8.7% were positive by rapid test and 12.9% had Gram stain scores >7. Rapid test sensitivity and specificity were 35.0% and 95.2%, respectively. Sensitivity was higher (76.3%) when considering only pH results, but specificity decreased (77.8%). CONCLUSION: The FemExam® is a specific but insensitive test for rapid diagnosis of BV in asymptotnatic pregnant women. The amines component appears to be limiting the test's overall sensitivity. To determine the more useful test tor rapid BV diagnosis, a comparison of the FemExam® to Amsel's criteria is needed and undenvay.

Table Diagnostic characteristics of rapid test compared to gram stain

SENSITMTY pH + and amines + pH + or amines + pH + Amines +

35.0% 82.5% 76.3% 41.3%

POSITIVE NEGATIVE PREDICTIVE PREDICTIVE SPECIFICITY VALUE VALUE 95.2% 69.6% 77.8% 87.0%

51.9% 28.7% 33.7% 32.0%

90.8% 96.4% 95.7% 90.9%