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Abruptio Placenta and preterm delivery: Thrombin effects on endometrial stromal cell prostaglandin expression
SPO Abstracts 403
Volume 172, N u m b e r l, Part 2 Am.] Obstet Gynecol
517 HEMATOLOGIC EVIDENCE THAT THE HUMAN FETUS
PARTICIPATES IN THE PROCESS LEADING TO PRETERM PARTURITION. R. Romero, S. Berry, R. Gomez% H. Mufioz', M. Mazor% Y. Sorokin=, F. Ghezzi*, D.B. Cotton. Wayne State University/Hutzel Hospital, Detroit, MI, Perinatology Research Branch, NICHD, Bethesda~ MD, Soroka Medical Center, Ben-Gnrion University, Iker-Sbeva, Israel. BACKGROUND/OBJECTIVE: The implicit paradigm governing the clinical management of preterm labor (PTL) is that this condition is a primary uterine/maternal disorder and benc¢ the jnstification for tocolytic treatment. Vet, across species, the fetus is an active participant in determining when labor should begin. We propose that PTL is the result of fetal disease and the fetus is actively involved in the process of preterm parturition. For this hypothesis to be correct, there must be significant biological differences between fetuses presenting with PTL who deliver preterm and those who deliver at term. We hypothesized that stressassociated hcmatologio changes in fetal blond will identify those fetuses who will deliver preterm. STUDY DESIGN: Cordocentesis/amniocentcsis were performed on 66 patients with preterm labor and intact membranes (22-34 weeks). The study was approved by the Institution Review Board and patients gave written consent. A complete fetal hematologic profile including red blood cell (RBC) count and indices, white blood cell (WBC) count with differential count and platelet count, was obtained. Results of these studies were not used for patient management decisions. Since hematologic values change with gestationsl age, results were expressed as the ratio of the observed/50th centile for gestationsl age. RESULTS: Fetuses who delivered preterm (< 72 h from cordocentesis) (n= 27) had significantly higher WBC, neutrophil (NE) and RBC count ratios than tbose who subseqaently delivered at term (n=39) (WBC: median: 1.41; range: 0.56-5.31 vs. 1.08; range: 0.44-2.91, respectively, p <0.05); NE ratio: 2.38 (range 0.31-20) vs. 1.24 (range 0.32-15.6, rcspoetively, p <0.05); RBC: 1.11; range: 0.78-1.36 vs. 0.3 range 0.68-1.4, respectively, p <0.05). No differences were found in platelet and RBC indices. The elevation in fetal WBC and NE ratio occurred even in patients WITHOUT microbial invasion of the amniotic cavity. CONCLUSION: Fetuses presenting with preterm labor destined to deliver prematurely are fundamentally different than those who will deliver at term.
518
~ C I g AND ~
OF ~ MATRIX D g g l A D I N G IINZIfM]gs INlllllrroR O1~l ~ l g r ~ ( Y i ' E i l q A , ~ g 8 fIIMP.1) IN llgrAL MI~IBRAIqlg. S.~.Fortunoto, R. Menon, ILF. Swan, W.ll. Barlcos. Dopta. of O b ~ and Biochemistry, 'rulane Uulve~ity Sdhcol of Medidne, New Orleans, La. To define the expression and synthesis of fetal membrane proteaute= and inhibitors Glmp-1) whida may pertain to PROM. STUDY DI~glGN: AmulodmHonle membrane= were collected from women who were undergoing normal elective C-sertion. Membranes were eat Into 5ram rirde= and maintained in an organ explant system for 48 hours. Culture media containing FBS was replaced with 0.2% lactalbumln hyd=rolysateand maintained for another 24 hours. Some tle=ue samples were stimulated with LPS (S0ng/ml) and recombinant IL-6 (10ng/ml). ' n ~ u e samples were frozen and or fixed at various time i~rloda for RT-PCR and in sttu hybridization. Media containing laetalbumin were frozen after the 24 hour incubation for zymogram studies. RT-PCR was done to study the expression of TIMP.I and interstitial collagenose. RESULT~ mRNA for TIMP-I was seen in amniodmrloule membrane at the time of collection and also during culture. Expre~ion of interstitial coUagenase mRNA was not seen in the nmnindmrion. LPS and recombinant IL-6 appeared to have no effect on the ezpre=don of TIMPq or in the induction of collagenase. In situ hybridization demonstrated that the message for TIMP-I was localized exclusively in the chorloule celia. Culture media were subjected to SDS*substrato gel eleetsophoreais (llelatinzymography) and showed band= at 72 and 92kD. The activity of t h ~ bands was completdly inhibited by 10raM EDTA sugg~ting that t h ~ banda were due to type IV coUagene=e (gehttinnse) activity. 11seue at the time of collection (not in culture) indicated the pre=ance of 72kD gelatimu~ (type IV collagcunse) demonstrating the indudion of only the 92kD form during colture.
CONCLUSION: These data infer that endogenous smniochorlon protenses may play a role in preterm PROM.
519 ABRUPT10 PLACENTA AND PRETERM DELIVERY:
THROMBIN EFFECTS ON ENDOMETRIAL STROMAL CELL PROSTAGLANDIN EXPRESSION C.J. Lockwood, Mt. Sinai School of Medicine, NY, NY. O B J E C T I V E : The pathogenic mechanism(s) underlying the association between preterm delivery (PTD) and decidual hemorrhage, (i.e., abmptio placenta), are unknown. Deeidual cells and endometrial stromal cells (ESCs) contain chronically high levels of tissue factor, the ultimate arbiter of thrombin (T) generation. In addition to its coagulant properties, T induces prostaglandin (Pg) expression in endothelial ceils. Thus, we determined whether T also enhanced Pg production by ESCs Lrlvitro, and whether T-effects were modified by steroids. METHODS: Confluent ESC cultures were exposed to vehicle control (ctr), 10-8M nstradiol (E2), 10-6M medroxyprogesterone acetate (MPA) or E2+MPA for 5 days, then exposed to the corresponding medium +2.3 U/ml o f t for 24 hours. This T coneentraton was based on initial dose response studies. The medium was then analyzed for PgE2, PgF2ct and their inactive metabolites by immunoassay. RESULTS: T significantly enhanced PgF2ct release in control and E2 but not MPA.-I:E2treated cultures (see Table). T did not affect the release of PgE2 or Pg metabolites in any of the culture conditions. mean (SEM) PgF2ot in pg]ml of media/ug culture protien (n=7) Ctr E2 MPA E2+MPA -T +T -T +T -T +T -T +T 31.7 58.7* 34.4 205.9* 33.1 44.8 49.1 71.5 (6.5) (9.8) (4.5) ~64.2) ( 6 . 0 ) ( 4 . 4 t (24.7) (24.1)
(* = P<0.5 for comparison of +T and -T exposed cultures) CONCLUSIONS: These findings suggest that physiological levels of thrombin augment cultured ESC PgF2e~expression without increasing its degradation, but only in a hypoprogestational milieu. Supported by NIH R29HD2950-02.
520 PRE-TERM DELIVERY WITH POLYIIYI)RAMNIOS MAY NOT BE DUE TO AN OVER DISTENDED UTERUS. A. Mann v*'x, N. Lazebnlk3, L. BLIP. Mage~Womens Hmp'ltal Research Inttltutet and the Dept Ob/Gyns, Uaiv of Pitt, P i t , b u r g h , PA. OBJECTIVE: To determine if there it an tameiation between the severity of polyhydramnins and the frequency of pre-term labor. STUDY DESIGN: The study group consisted of 275 singleton pregnon¢ies with polyhydramninsas defined by an anmioti¢ fluid index (AFI) of > 25 cm. Polyhydrarnnins was defined as mild (AFI: 25-30 era); moderate (AFI: 30.1-35.0 era) and severe (AFI ~ 35.1 cm). A delivery was considered pre-term if it occurred prior to 37.0 weeks'
ge~tatinn. RESULTS: 55/275 (20%) newborns delivered before 37.0 weeks' gestation. The ineldcoee of pgetonn delivery watt not signlftcantly different bgtweco patients with mild (38/199; 19~); moderste (14/55; 25%); and severe (3/21; 14%) polyhydramnios. Among patlenta with idiopathic polyhydramnins, the incidence=of preterm delivery (28/190; 14%) was not significantly different from the overall rate of pre-term deliveries for singleton pregnancle= at our hospital (1053/8755; 12.2%). However, fetuses with congenital malformations (14/41; 35%) and women with geztalionsl or inanlin-dependent diabet~ mellitus (14/49; 28.5%) had • signifgantly higher incidence of preterm delivery (p < 0.04) than those fetuses with idiopathic polyhydranmins, regardlnss of severity, CONCLUSION: The ~'verity of polyhydranmins doe= not correlate with the incidence of pre-tcrm delivery. The underlying cause of polyhydranmios, rather than the relative amount of amnintin fluid as defined by this study, appeara to affect when labor will occur.
Volume 172, N u m b e r l, Part 2 Am.] Obstet Gynecol
517 HEMATOLOGIC EVIDENCE THAT THE HUMAN FETUS
PARTICIPATES IN THE PROCESS LEADING TO PRETERM PARTURITION. R. Romero, S. Berry, R. Gomez% H. Mufioz', M. Mazor% Y. Sorokin=, F. Ghezzi*, D.B. Cotton. Wayne State University/Hutzel Hospital, Detroit, MI, Perinatology Research Branch, NICHD, Bethesda~ MD, Soroka Medical Center, Ben-Gnrion University, Iker-Sbeva, Israel. BACKGROUND/OBJECTIVE: The implicit paradigm governing the clinical management of preterm labor (PTL) is that this condition is a primary uterine/maternal disorder and benc¢ the jnstification for tocolytic treatment. Vet, across species, the fetus is an active participant in determining when labor should begin. We propose that PTL is the result of fetal disease and the fetus is actively involved in the process of preterm parturition. For this hypothesis to be correct, there must be significant biological differences between fetuses presenting with PTL who deliver preterm and those who deliver at term. We hypothesized that stressassociated hcmatologio changes in fetal blond will identify those fetuses who will deliver preterm. STUDY DESIGN: Cordocentesis/amniocentcsis were performed on 66 patients with preterm labor and intact membranes (22-34 weeks). The study was approved by the Institution Review Board and patients gave written consent. A complete fetal hematologic profile including red blood cell (RBC) count and indices, white blood cell (WBC) count with differential count and platelet count, was obtained. Results of these studies were not used for patient management decisions. Since hematologic values change with gestationsl age, results were expressed as the ratio of the observed/50th centile for gestationsl age. RESULTS: Fetuses who delivered preterm (< 72 h from cordocentesis) (n= 27) had significantly higher WBC, neutrophil (NE) and RBC count ratios than tbose who subseqaently delivered at term (n=39) (WBC: median: 1.41; range: 0.56-5.31 vs. 1.08; range: 0.44-2.91, respectively, p <0.05); NE ratio: 2.38 (range 0.31-20) vs. 1.24 (range 0.32-15.6, rcspoetively, p <0.05); RBC: 1.11; range: 0.78-1.36 vs. 0.3 range 0.68-1.4, respectively, p <0.05). No differences were found in platelet and RBC indices. The elevation in fetal WBC and NE ratio occurred even in patients WITHOUT microbial invasion of the amniotic cavity. CONCLUSION: Fetuses presenting with preterm labor destined to deliver prematurely are fundamentally different than those who will deliver at term.
518
~ C I g AND ~
OF ~ MATRIX D g g l A D I N G IINZIfM]gs INlllllrroR O1~l ~ l g r ~ ( Y i ' E i l q A , ~ g 8 fIIMP.1) IN llgrAL MI~IBRAIqlg. S.~.Fortunoto, R. Menon, ILF. Swan, W.ll. Barlcos. Dopta. of O b ~ and Biochemistry, 'rulane Uulve~ity Sdhcol of Medidne, New Orleans, La. To define the expression and synthesis of fetal membrane proteaute= and inhibitors Glmp-1) whida may pertain to PROM. STUDY DI~glGN: AmulodmHonle membrane= were collected from women who were undergoing normal elective C-sertion. Membranes were eat Into 5ram rirde= and maintained in an organ explant system for 48 hours. Culture media containing FBS was replaced with 0.2% lactalbumln hyd=rolysateand maintained for another 24 hours. Some tle=ue samples were stimulated with LPS (S0ng/ml) and recombinant IL-6 (10ng/ml). ' n ~ u e samples were frozen and or fixed at various time i~rloda for RT-PCR and in sttu hybridization. Media containing laetalbumin were frozen after the 24 hour incubation for zymogram studies. RT-PCR was done to study the expression of TIMP.I and interstitial collagenose. RESULT~ mRNA for TIMP-I was seen in amniodmrloule membrane at the time of collection and also during culture. Expre~ion of interstitial coUagenase mRNA was not seen in the nmnindmrion. LPS and recombinant IL-6 appeared to have no effect on the ezpre=don of TIMPq or in the induction of collagenase. In situ hybridization demonstrated that the message for TIMP-I was localized exclusively in the chorloule celia. Culture media were subjected to SDS*substrato gel eleetsophoreais (llelatinzymography) and showed band= at 72 and 92kD. The activity of t h ~ bands was completdly inhibited by 10raM EDTA sugg~ting that t h ~ banda were due to type IV coUagene=e (gehttinnse) activity. 11seue at the time of collection (not in culture) indicated the pre=ance of 72kD gelatimu~ (type IV collagcunse) demonstrating the indudion of only the 92kD form during colture.
CONCLUSION: These data infer that endogenous smniochorlon protenses may play a role in preterm PROM.
519 ABRUPT10 PLACENTA AND PRETERM DELIVERY:
THROMBIN EFFECTS ON ENDOMETRIAL STROMAL CELL PROSTAGLANDIN EXPRESSION C.J. Lockwood, Mt. Sinai School of Medicine, NY, NY. O B J E C T I V E : The pathogenic mechanism(s) underlying the association between preterm delivery (PTD) and decidual hemorrhage, (i.e., abmptio placenta), are unknown. Deeidual cells and endometrial stromal cells (ESCs) contain chronically high levels of tissue factor, the ultimate arbiter of thrombin (T) generation. In addition to its coagulant properties, T induces prostaglandin (Pg) expression in endothelial ceils. Thus, we determined whether T also enhanced Pg production by ESCs Lrlvitro, and whether T-effects were modified by steroids. METHODS: Confluent ESC cultures were exposed to vehicle control (ctr), 10-8M nstradiol (E2), 10-6M medroxyprogesterone acetate (MPA) or E2+MPA for 5 days, then exposed to the corresponding medium +2.3 U/ml o f t for 24 hours. This T coneentraton was based on initial dose response studies. The medium was then analyzed for PgE2, PgF2ct and their inactive metabolites by immunoassay. RESULTS: T significantly enhanced PgF2ct release in control and E2 but not MPA.-I:E2treated cultures (see Table). T did not affect the release of PgE2 or Pg metabolites in any of the culture conditions. mean (SEM) PgF2ot in pg]ml of media/ug culture protien (n=7) Ctr E2 MPA E2+MPA -T +T -T +T -T +T -T +T 31.7 58.7* 34.4 205.9* 33.1 44.8 49.1 71.5 (6.5) (9.8) (4.5) ~64.2) ( 6 . 0 ) ( 4 . 4 t (24.7) (24.1)
(* = P<0.5 for comparison of +T and -T exposed cultures) CONCLUSIONS: These findings suggest that physiological levels of thrombin augment cultured ESC PgF2e~expression without increasing its degradation, but only in a hypoprogestational milieu. Supported by NIH R29HD2950-02.
520 PRE-TERM DELIVERY WITH POLYIIYI)RAMNIOS MAY NOT BE DUE TO AN OVER DISTENDED UTERUS. A. Mann v*'x, N. Lazebnlk3, L. BLIP. Mage~Womens Hmp'ltal Research Inttltutet and the Dept Ob/Gyns, Uaiv of Pitt, P i t , b u r g h , PA. OBJECTIVE: To determine if there it an tameiation between the severity of polyhydramnins and the frequency of pre-term labor. STUDY DESIGN: The study group consisted of 275 singleton pregnon¢ies with polyhydramninsas defined by an anmioti¢ fluid index (AFI) of > 25 cm. Polyhydrarnnins was defined as mild (AFI: 25-30 era); moderate (AFI: 30.1-35.0 era) and severe (AFI ~ 35.1 cm). A delivery was considered pre-term if it occurred prior to 37.0 weeks'
ge~tatinn. RESULTS: 55/275 (20%) newborns delivered before 37.0 weeks' gestation. The ineldcoee of pgetonn delivery watt not signlftcantly different bgtweco patients with mild (38/199; 19~); moderste (14/55; 25%); and severe (3/21; 14%) polyhydramnios. Among patlenta with idiopathic polyhydramnins, the incidence=of preterm delivery (28/190; 14%) was not significantly different from the overall rate of pre-term deliveries for singleton pregnancle= at our hospital (1053/8755; 12.2%). However, fetuses with congenital malformations (14/41; 35%) and women with geztalionsl or inanlin-dependent diabet~ mellitus (14/49; 28.5%) had • signifgantly higher incidence of preterm delivery (p < 0.04) than those fetuses with idiopathic polyhydranmins, regardlnss of severity, CONCLUSION: The ~'verity of polyhydranmins doe= not correlate with the incidence of pre-tcrm delivery. The underlying cause of polyhydranmios, rather than the relative amount of amnintin fluid as defined by this study, appeara to affect when labor will occur.