- Email: [email protected]
Normal pregnancy is associated with the development of protein S and protein Z antibodies, independent of PS and PZ levels
S140 SMFM Abstracts 488
ENDOCRINE DISRUPTORS IN THE MATERNAL AND FETAL COMPARTMENTS SUSAN LASHLEY1, ANTONIA CALAFAT2, DANA BARR2, THOMAS LEDOUX3, PAROMITA HORE4, MARIAN LAKE1, MARK ROBSON4, JOHN SMULIAN1, 1UMDNJRobert Wood Johnson Medical School/Robert Wood Johnson University Hospital, Obstetrics, Gynecology and Reproductive Sciences, New Brunswick, New Jersey, 2Centers for Disease Control and Prevention (CDC), Division of Laboratory Sciences, National Center for Environmental Health, Atlanta, Georgia, 3New Jersey Department of Environmental Protection, Risk Assessment & Toxicology Section, Division of Science and Research, Trenton, New Jersey, 4University of Medicine and Dentistry of New Jersey, School of Public Health, Dept. of Environmental & Occupational Health, Piscataway, New Jersey OBJECTIVE: Phthalates (plasticizers) are known endocrine disruptors in the animal kingdom and are associated with developmental toxicities. This study was designed to describe the relationship between levels of phthalate metabolites in human maternal serum and cord serum. STUDY DESIGN: Cord blood and maternal blood were collected from 50 maternal-fetal pairs at the time of elective cesarean section between July 2003May 2004. The collection materials were tested prior to obtaining samples and the collection procedure was designed to minimize the potential for phthalate contamination. The Centers for Disease Control and Prevention analyzed the maternal and fetal sera, using automated solid phase extraction-isotope dilutionhigh performance liquid chromatography-tandem mass spectrometry, for nine metabolites of phthalates. Correlation of phthalates in maternal and fetal sera was assessed based on spearman rank correlation coefficient (r). RESULTS: Phthalate metabolites were detected in 100% of maternal and cord serum samples. However, the frequency of detection for each specific metabolite differed between the two compartments. Seven of the nine phthalate metabolites were more frequently detected in cord serum than maternal serum. For example, mono-2-ethylhexyl phthalate (MEHP) was detected in all of the cord samples but in only 69% of maternal samples (P ! .001). The concentration of MEHP, mono-(2-ethyl-5-hydroxyhexyl) phthalate, and mono-(2-ethyl-5-oxohexyl) phthalate were higher in cord serum then the maternal serum. When metabolites were present in both compartments, a correlation between levels in maternal and fetal sera was found for mono-n-butyl phthalate P = .03 (r =.54), mono-benzyl phthalate P = .04 (r =.40), mono-ethyl phthalate P = .007 (r =.41). CONCLUSION: This study confirms that phthalates metabolites are present in the human fetal compartment. The metabolites levels in cord serum were generally higher than in maternal serum. These results may indicate that the mechanism of fetal clearance of phthalates may predispose to trapping of phthalates in the fetal compartment.
490
HEAT SHOCK PROTEIN-70: AN UNEXPECTED DECREASE IN A STRESS-DETERMINANT IN NEW YORK CITY GRAVIDAS AFTER THE EVENTS OF SEPTEMBER 11, 2001 MENJEAN LEE1, NIKKI KOKLANARIS1, LORRAINE O’NEILL1, MANISH SHAH1, MORTIMER LEVITZ1, NAIMA ISMAILI2, MICHAEL GARABEDIAN2, 1New York University, Obstetrics and Gynecology, New York, New York, 2New York University, Microbiology, New York, New York OBJECTIVE: The purpose of this study was to evaluate the levels of adrenocorticotropin hormone (ACTH), cortisol, and heat shock protein-70 (HSP-70) in mid-gestation gravidas before and after the events of September 11, 2001. STUDY DESIGN: This was a nested cohort study of banked maternal serum samples drawn at approximately 20 weeks of gestational age from women with low-risk pregnancies. The pre-9/11 samples were drawn in the year prior to 9/11; the post-9/11 samples were drawn within 6 months after the crisis. The pre-9/11 and post-9/11 samples were matched 1:1 on the basis of age, race, and parity.The serum samples were assayed for levels of ACTH and cortisol by a commercially available ImmuliteÔ system. HSP-70 levels, an additional marker for stress, were measured by a commercially available ELISA. T-tests were performed to examine any differences in randomly drawn stress hormone levels and HSP-70. RESULTS: There were no statistical differences in the mean values of randomly drawn ACTH and cortisol levels between the groups. However, the mean values of HSP-70 were significantly lower in the post-9/11 group: 15.950 pg/ml (G8.9 SEM) in the post-9/11 group vs 122.25 pg/ml (G31.3 SEM) in the pre-9/11 group (P = .0023). CONCLUSION: Unexpectedly, a marker for cellular stress was decreased in the gravidas who experienced the events of 9/11. In the animal model, pregnancy conveys an altered response to external stressors: the neuroendocrine response to stimuli irrelevant to reproduction has been noted to be down-regulated in the pregnant rat. Our data suggest that HSP-70 may play a role in mediating a similar response in humans.
489
OXIDATIVE STRESS AND INTRAUTERINE GROWTH RESTRICTION (IUGR): A SELECTIVE INABILITY OF THE FETUS TO RESPOND TO AN OXIDATIVE INSULT VISWANATHAN RAVISHANKAR1, ERROL NORWITZ1, CATALIN BUHIMSCHI1, CARMEN BOOTH2, JOSHUA COPEL1, IRINA BUHIMSCHI1, 1Yale University, Ob./ Gyn. & Repro. Sci., New Haven, Connecticut, 2Yale University, Comparative Medicine, New Haven, Connecticut OBJECTIVE: IUGR is a major cause of perinatal morbidity and mortality and its cause remains unknown. We hypothesize IUGR results from the inability of the fetus to compensate in a setting of increased oxidative stress. This study explores the ability of the mother, fetus and placenta to adapt to an increased oxidative stress induced by chronic hypoxia and nitric oxide synthase (NOS) inhibition. STUDY DESIGN: Timed pregnant Sprague-Dawley rats were acquired on d17 of gestation (term = d22). L-NAME (NOS inhibitor) infusion pumps were inserted on d18. Control animals received saline. Study groups: (1) normoxia+ saline (20%S: n = 7); (2) normoxia+L-NAME (20%LN, n = 8); (3) 10% hypoxia+saline (10%S, n = 6); and (4) 10% hypoxia+L-NAME (10%LN: n = 6). Hypoxia was induced by housing in a hypoxic chamber according to their group allocation and sacrificed on day 21. Maternal weight, litter size, and pup weights were measured. Glutathione (GSH) and glutathione disulfide (GSSG) levels were measured in the maternal, placental tissue, and fetal circulation. RESULTS: Maternal and fetal weights were significantly decreased in the challenged group (P ! .001). Oxidative insult resulted in increased maternal blood levels of GSH (10%LN: 1229.0 G 78.8 vs. 20%S: 954.6 G 67.0 mM [P ! .05], GSSG (10%LN: 12.1 G 2.1 vs. 1.5 G 0.6 mM [P ! .05] but a decrease in GSH / GSSG ratio (10%LN: 113.1 G 18.1 vs. 20%S: 1329.7 G 351.4 [P ! .05]). Similar results were noted in placental tissues. In contrast, the oxidative insult resulted in decrease fetal blood levels of GSH (10%LN: 763.5 G 48.5 vs. 20%S: 1109.2 G 48.9 mM [P ! .05]) and GSSG concentrations (10%LN: 0.8 G 0.4 vs 20%S: 2.1 G 0.8 mM [P ! .05]) and no significant difference in GSH / GSSG ratio (10%LN: 2922.0 G 1356.7 vs 20%S: 2304.0 G 874.1 [P O .05]). CONCLUSION: Hypoxia and NOS inhibition induces a synergistic oxidative stress to which the mother responds with increased circulating levels of antioxidants (GSH). This most probably represents a compensatory defense mechanism against excess free radicals. In contrast, but the fetus is not able to activate such mechanisms and responds with IUGR.
491
NORMAL PREGNANCY IS ASSOCIATED WITH THE DEVELOPMENT OF PROTEIN S AND PROTEIN Z ANTIBODIES, INDEPENDENT OF PS AND PZ LEVELS MICHAEL PAIDAS1, DE-HUI KU2, YALE ARKEL2, ELIZABETH TRICHE3, CHRISTINE FORTUNATO2, BEN HAMAR2, EVELYN KU2, CHARLES LOCKWOOD2, 1Yale University, Obsteteric, Gyencology and Reproductive Sciences, New Haven, Connecticut, 2Yale University, Obstetrics, Gynecology and Reproductive Sciences, New Haven, Connecticut, 3Yale University, Epidemiology and Public Health, New Haven, Connecticut OBJECTIVE: Protein S (PS) and Protein Z (PZ) are involved in regulating thrombin. We sought to determine whether normal pregnancy (NP) is associated with the development of anti-PS, and anti- PZ antibodies (Ab), and whether these Ab, if present, correlate with PS and PZ levels. STUDY DESIGN: We measured maternal PZ, free PS, anti-IgG PS, anti-IgM PS, anti-PZ IgG, anti-PZ IgM Ab in 1st, 2nd, and 3rd trimester (TRI) from healthy women (n = 82 in eachTRI) with subsequent NP outcome. Reference (Ref) values were based upon data from non-pregnant adults (n) for free PS (68), PZ (104), anti-PS anti- PZ IgG and IgM Ab (50). Free PS, PZ, anti-PS and -PZ IgG and IgM Ab were measured by ELISA [we created ELISA for PS and PZ Ab; levels expressed in optical density (OD)]. Data analysis was performed using Pearson correlation (r) and t test for single samples (sig P ! .05). RESULTS: All three TRI were associated with significantly different values compared to Ref means (P ! .05, t test), except for data expressed with asterisk (Table). Anti- PS IgM correlated with anti-PZ IgM antibodies in 1st and 3rd TRI (r 0.43, 0.46 resp, P ! .05), while the 2nd TRI was borderline (r 0.20). AntiPS IgG correlated with anti-PZ IgG antibodies in 3rd TRI (r 0.29, P ! .05), but not in the 1st or 2nd TRI. There was no correlation between PS levels and antiPS IgG or anti- PS IgM in any TRI, except for PS and anti-PS IgG in the 2nd TRI (r 0.25, P ! .05). There was no correlation between PZ levels and anti-PZ IgG, while 2nd and 3rd TRI PZ levels correlated with anti-PZ IgM (r 0.26 and ÿ0.28 resp, P ! .05).
Test
Ref Mean
Ref SD
1st tri Mean
1st tri SD
2nd tri Mean
2nd tri SD
3rd tri Mean
3rd tri SD
PZ ug/mL Free PS % PZ IgG OD PZ IgM OD PS IgG OD PS IgM OD
1.56 88 0.19 0.07 0.27 0.26
0.61 19 0.14 0.17 0.20 0.15
2.54 39 0.26* 0.36 0.29* 0.48
0.87 10.46 0.41 0.22 0.35 0.38
2 35.6 0.19* 0.21 0.44 0.63
0.66 8.42 0.28 0.11 0.24 0.41
2.14 27.9 0.3 0.38 0.44 0.61
0.85 6.98 0.28 0.2 0.30 0.36
CONCLUSION: NP is associated with the development of PS and PZ antibodies as gestation progresses, independent of PZ and PS levels, and may contribute to the prothrombotic milieu of pregnancy.
ENDOCRINE DISRUPTORS IN THE MATERNAL AND FETAL COMPARTMENTS SUSAN LASHLEY1, ANTONIA CALAFAT2, DANA BARR2, THOMAS LEDOUX3, PAROMITA HORE4, MARIAN LAKE1, MARK ROBSON4, JOHN SMULIAN1, 1UMDNJRobert Wood Johnson Medical School/Robert Wood Johnson University Hospital, Obstetrics, Gynecology and Reproductive Sciences, New Brunswick, New Jersey, 2Centers for Disease Control and Prevention (CDC), Division of Laboratory Sciences, National Center for Environmental Health, Atlanta, Georgia, 3New Jersey Department of Environmental Protection, Risk Assessment & Toxicology Section, Division of Science and Research, Trenton, New Jersey, 4University of Medicine and Dentistry of New Jersey, School of Public Health, Dept. of Environmental & Occupational Health, Piscataway, New Jersey OBJECTIVE: Phthalates (plasticizers) are known endocrine disruptors in the animal kingdom and are associated with developmental toxicities. This study was designed to describe the relationship between levels of phthalate metabolites in human maternal serum and cord serum. STUDY DESIGN: Cord blood and maternal blood were collected from 50 maternal-fetal pairs at the time of elective cesarean section between July 2003May 2004. The collection materials were tested prior to obtaining samples and the collection procedure was designed to minimize the potential for phthalate contamination. The Centers for Disease Control and Prevention analyzed the maternal and fetal sera, using automated solid phase extraction-isotope dilutionhigh performance liquid chromatography-tandem mass spectrometry, for nine metabolites of phthalates. Correlation of phthalates in maternal and fetal sera was assessed based on spearman rank correlation coefficient (r). RESULTS: Phthalate metabolites were detected in 100% of maternal and cord serum samples. However, the frequency of detection for each specific metabolite differed between the two compartments. Seven of the nine phthalate metabolites were more frequently detected in cord serum than maternal serum. For example, mono-2-ethylhexyl phthalate (MEHP) was detected in all of the cord samples but in only 69% of maternal samples (P ! .001). The concentration of MEHP, mono-(2-ethyl-5-hydroxyhexyl) phthalate, and mono-(2-ethyl-5-oxohexyl) phthalate were higher in cord serum then the maternal serum. When metabolites were present in both compartments, a correlation between levels in maternal and fetal sera was found for mono-n-butyl phthalate P = .03 (r =.54), mono-benzyl phthalate P = .04 (r =.40), mono-ethyl phthalate P = .007 (r =.41). CONCLUSION: This study confirms that phthalates metabolites are present in the human fetal compartment. The metabolites levels in cord serum were generally higher than in maternal serum. These results may indicate that the mechanism of fetal clearance of phthalates may predispose to trapping of phthalates in the fetal compartment.
490
HEAT SHOCK PROTEIN-70: AN UNEXPECTED DECREASE IN A STRESS-DETERMINANT IN NEW YORK CITY GRAVIDAS AFTER THE EVENTS OF SEPTEMBER 11, 2001 MENJEAN LEE1, NIKKI KOKLANARIS1, LORRAINE O’NEILL1, MANISH SHAH1, MORTIMER LEVITZ1, NAIMA ISMAILI2, MICHAEL GARABEDIAN2, 1New York University, Obstetrics and Gynecology, New York, New York, 2New York University, Microbiology, New York, New York OBJECTIVE: The purpose of this study was to evaluate the levels of adrenocorticotropin hormone (ACTH), cortisol, and heat shock protein-70 (HSP-70) in mid-gestation gravidas before and after the events of September 11, 2001. STUDY DESIGN: This was a nested cohort study of banked maternal serum samples drawn at approximately 20 weeks of gestational age from women with low-risk pregnancies. The pre-9/11 samples were drawn in the year prior to 9/11; the post-9/11 samples were drawn within 6 months after the crisis. The pre-9/11 and post-9/11 samples were matched 1:1 on the basis of age, race, and parity.The serum samples were assayed for levels of ACTH and cortisol by a commercially available ImmuliteÔ system. HSP-70 levels, an additional marker for stress, were measured by a commercially available ELISA. T-tests were performed to examine any differences in randomly drawn stress hormone levels and HSP-70. RESULTS: There were no statistical differences in the mean values of randomly drawn ACTH and cortisol levels between the groups. However, the mean values of HSP-70 were significantly lower in the post-9/11 group: 15.950 pg/ml (G8.9 SEM) in the post-9/11 group vs 122.25 pg/ml (G31.3 SEM) in the pre-9/11 group (P = .0023). CONCLUSION: Unexpectedly, a marker for cellular stress was decreased in the gravidas who experienced the events of 9/11. In the animal model, pregnancy conveys an altered response to external stressors: the neuroendocrine response to stimuli irrelevant to reproduction has been noted to be down-regulated in the pregnant rat. Our data suggest that HSP-70 may play a role in mediating a similar response in humans.
489
OXIDATIVE STRESS AND INTRAUTERINE GROWTH RESTRICTION (IUGR): A SELECTIVE INABILITY OF THE FETUS TO RESPOND TO AN OXIDATIVE INSULT VISWANATHAN RAVISHANKAR1, ERROL NORWITZ1, CATALIN BUHIMSCHI1, CARMEN BOOTH2, JOSHUA COPEL1, IRINA BUHIMSCHI1, 1Yale University, Ob./ Gyn. & Repro. Sci., New Haven, Connecticut, 2Yale University, Comparative Medicine, New Haven, Connecticut OBJECTIVE: IUGR is a major cause of perinatal morbidity and mortality and its cause remains unknown. We hypothesize IUGR results from the inability of the fetus to compensate in a setting of increased oxidative stress. This study explores the ability of the mother, fetus and placenta to adapt to an increased oxidative stress induced by chronic hypoxia and nitric oxide synthase (NOS) inhibition. STUDY DESIGN: Timed pregnant Sprague-Dawley rats were acquired on d17 of gestation (term = d22). L-NAME (NOS inhibitor) infusion pumps were inserted on d18. Control animals received saline. Study groups: (1) normoxia+ saline (20%S: n = 7); (2) normoxia+L-NAME (20%LN, n = 8); (3) 10% hypoxia+saline (10%S, n = 6); and (4) 10% hypoxia+L-NAME (10%LN: n = 6). Hypoxia was induced by housing in a hypoxic chamber according to their group allocation and sacrificed on day 21. Maternal weight, litter size, and pup weights were measured. Glutathione (GSH) and glutathione disulfide (GSSG) levels were measured in the maternal, placental tissue, and fetal circulation. RESULTS: Maternal and fetal weights were significantly decreased in the challenged group (P ! .001). Oxidative insult resulted in increased maternal blood levels of GSH (10%LN: 1229.0 G 78.8 vs. 20%S: 954.6 G 67.0 mM [P ! .05], GSSG (10%LN: 12.1 G 2.1 vs. 1.5 G 0.6 mM [P ! .05] but a decrease in GSH / GSSG ratio (10%LN: 113.1 G 18.1 vs. 20%S: 1329.7 G 351.4 [P ! .05]). Similar results were noted in placental tissues. In contrast, the oxidative insult resulted in decrease fetal blood levels of GSH (10%LN: 763.5 G 48.5 vs. 20%S: 1109.2 G 48.9 mM [P ! .05]) and GSSG concentrations (10%LN: 0.8 G 0.4 vs 20%S: 2.1 G 0.8 mM [P ! .05]) and no significant difference in GSH / GSSG ratio (10%LN: 2922.0 G 1356.7 vs 20%S: 2304.0 G 874.1 [P O .05]). CONCLUSION: Hypoxia and NOS inhibition induces a synergistic oxidative stress to which the mother responds with increased circulating levels of antioxidants (GSH). This most probably represents a compensatory defense mechanism against excess free radicals. In contrast, but the fetus is not able to activate such mechanisms and responds with IUGR.
491
NORMAL PREGNANCY IS ASSOCIATED WITH THE DEVELOPMENT OF PROTEIN S AND PROTEIN Z ANTIBODIES, INDEPENDENT OF PS AND PZ LEVELS MICHAEL PAIDAS1, DE-HUI KU2, YALE ARKEL2, ELIZABETH TRICHE3, CHRISTINE FORTUNATO2, BEN HAMAR2, EVELYN KU2, CHARLES LOCKWOOD2, 1Yale University, Obsteteric, Gyencology and Reproductive Sciences, New Haven, Connecticut, 2Yale University, Obstetrics, Gynecology and Reproductive Sciences, New Haven, Connecticut, 3Yale University, Epidemiology and Public Health, New Haven, Connecticut OBJECTIVE: Protein S (PS) and Protein Z (PZ) are involved in regulating thrombin. We sought to determine whether normal pregnancy (NP) is associated with the development of anti-PS, and anti- PZ antibodies (Ab), and whether these Ab, if present, correlate with PS and PZ levels. STUDY DESIGN: We measured maternal PZ, free PS, anti-IgG PS, anti-IgM PS, anti-PZ IgG, anti-PZ IgM Ab in 1st, 2nd, and 3rd trimester (TRI) from healthy women (n = 82 in eachTRI) with subsequent NP outcome. Reference (Ref) values were based upon data from non-pregnant adults (n) for free PS (68), PZ (104), anti-PS anti- PZ IgG and IgM Ab (50). Free PS, PZ, anti-PS and -PZ IgG and IgM Ab were measured by ELISA [we created ELISA for PS and PZ Ab; levels expressed in optical density (OD)]. Data analysis was performed using Pearson correlation (r) and t test for single samples (sig P ! .05). RESULTS: All three TRI were associated with significantly different values compared to Ref means (P ! .05, t test), except for data expressed with asterisk (Table). Anti- PS IgM correlated with anti-PZ IgM antibodies in 1st and 3rd TRI (r 0.43, 0.46 resp, P ! .05), while the 2nd TRI was borderline (r 0.20). AntiPS IgG correlated with anti-PZ IgG antibodies in 3rd TRI (r 0.29, P ! .05), but not in the 1st or 2nd TRI. There was no correlation between PS levels and antiPS IgG or anti- PS IgM in any TRI, except for PS and anti-PS IgG in the 2nd TRI (r 0.25, P ! .05). There was no correlation between PZ levels and anti-PZ IgG, while 2nd and 3rd TRI PZ levels correlated with anti-PZ IgM (r 0.26 and ÿ0.28 resp, P ! .05).
Test
Ref Mean
Ref SD
1st tri Mean
1st tri SD
2nd tri Mean
2nd tri SD
3rd tri Mean
3rd tri SD
PZ ug/mL Free PS % PZ IgG OD PZ IgM OD PS IgG OD PS IgM OD
1.56 88 0.19 0.07 0.27 0.26
0.61 19 0.14 0.17 0.20 0.15
2.54 39 0.26* 0.36 0.29* 0.48
0.87 10.46 0.41 0.22 0.35 0.38
2 35.6 0.19* 0.21 0.44 0.63
0.66 8.42 0.28 0.11 0.24 0.41
2.14 27.9 0.3 0.38 0.44 0.61
0.85 6.98 0.28 0.2 0.30 0.36
CONCLUSION: NP is associated with the development of PS and PZ antibodies as gestation progresses, independent of PZ and PS levels, and may contribute to the prothrombotic milieu of pregnancy.