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Prediction of abnormal pregnancy by trophoblast assessment in first trimester transcervical samples
(n¼1874) in which only embryos with minimal or no fragmentation (0-5%) were replaced were selected as control. Cycle outcomes were evaluated and compared. RESULTS: Pregnancy was confirmed in 13/43 (30%) patients with exclusive transfer of extensively fragmented embryos. Of these, seven pregnancies were biochemical, three were lost between 7 and 9 weeks gestation, and three resulted in birth, giving a live birth rate of 7% per cycle (3/43). All three infants had normal weights and were in apparently good health at birth. Pregnancy rate in the control group was 61.6% (1154/1874) per cycle. The live birth rate in this group was 44.8% (839/1874). Pregnancy loss rate following three consecutive rises in the beta hCG level was therefore 77% (10/13) in the study population and 27% (315/1154) in the control group. The average age in the two groups was not statistically different and the age range was similar. CONCLUSIONS: Implantation failure and early pregnancy loss are the most common outcomes after exclusive transfer of extensively fragmented embryos. This is in sharp contrast to the outcome of transfer when the embryos have minimal fragmentation: a large majority of the latter lead to live birth. Considering the risk of physical and psychological trauma associated with an aborted pregnancy, even in the absence of birth defects, patients for whom only embryos with extensive fragmentation are available should be counseled and warned of the high likelihood of a negative outcome following transfer. Supported by: Private.
P-10 PREDICTION OF ABNORMAL PREGNANCY BY TROPHOBLAST ASSESSMENT IN FIRST TRIMESTER TRANSCERVICAL SAMPLES. A. N. Imudia, Y. Suzuki, F. D. Yelian, M. P. Diamond, R. Romero, D. R. Armant. Obstetrics and Gynecology, Wayne State University, Detroit, MI; Perinatology Research Branch, Bethesda, MD; Reproductive Biology & Medicine Branch, Bethesda, MD. OBJECTIVE: Ectopic pregnancy complicates about 1-2% of all pregnancies and occurs when the developing blastocyst implants at a site other than the fundus of the uterine cavity, most commonly in the fallopian tube. Delayed clinical diagnosis of this abnormality can result in a dismal maternal outcome. The presence of fetal trophoblast cells in the cervical canal during the first trimester suggested a non-invasive approach for predicting abnormal pregnancy through transcervical sampling. DESIGN: Experimental pilot study. MATERIALS AND METHODS: This study was approved by the Wayne State University IRB and written informed consent was obtained from participating women. Patients, age 18-40, presenting for normal early prenatal care (n¼10), ectopic pregnancy (EP; n¼7), or blighted ovum (BO; n¼5) were enrolled for collection of transcervical specimens using a cytological brush and a ThinPrepÒ kit (Hologic). Cells collected in PreservCytÒ fixative solution were cleared of mucus by acidification, washed by centrifugation and an aliquot was prepared on a microscope slide using a Cytospin3 centrifuge. Slides were labeled with monoclonal antibody G233 recognizing HLA-G, a MHC antigen specifically expressed by trophoblast cells. All HLA-G positive cells were identified and counted on each slide. After staining with hematoxylin, the total number of cells on each slide was determined and the ratio of HLA-G positive cells to total cells was calculated. Data were compared using one-way ANOVA, the Student-Newman-Keuls posthoc test and receiver operating characteristic analysis (ROC). RESULTS: The mean gestational ages of normal intrauterine pregnancy (IUP), EP, and BO were 9, 8, and 10 weeks, respectively. Trophoblast cells were observed in 10/10 normal IUP, 3/7 EP, and 4/5 BO specimens. The frequency of HLA-G positive cells in the IUP cervical samples was approximately 1 in 2000, which was 5- to 10-fold higher (p<0.001) than the average frequency in samples from patients with EP or BO. Significantly, the identification of trophoblast cells in cervical samples demonstrated both 100% sensitivity and 100% specificity. CONCLUSIONS: Based on limited observations, trophoblast cells can be reliably identified among transcervical cells in the first trimester by immunohistochemical staining for HLA-G. This non-invasive procedure may be useful for prenatal diagnosis of pregnancy outcomes and could be adapted for genetic analysis, but will require further clinical investigation to establish its efficacy. Supported by: The intramural research program of NICHD.
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Abstracts
P-11 THE POLYCYSTIC OVARY SYNDROME IN JAPANESE RECURRENT MISCARRIAGE PATIENTS. M. Sugiura-Ogasawara, Y. Ozaki, T. Nakanishi, K. Nozawa, S. Obayashi. Obstetrics and Gynecology, Nagoya City University Medical School, Nagoya, Aichi, Japan; Nagoya City University Medical School, Nagoya, Japan. OBJECTIVE: The polycystic ovary syndrome (PCOS) is well-known to be associated with miscarriages. However, it has been a matter of controversy whether it is actually a causal factor because of the absence of internationally established criteria. We therefore here investigated whether PCOS and a polycystic ovary (PCO) morphology have predictive value for subsequent miscarriages using new International and Japanese criteria. DESIGN: Prospective study. MATERIALS AND METHODS: A total of 195 patients with a history of two consecutive first trimester miscarriages and no abnormal chromosomes in either partner, antiphospholipid antibodies or uterine anomalies, were examined. The prospective pregnancy outcome was compared between patients with and without PCOS, PCO morphology, elevated LH, hyperandrogenism, and obesity. RESULTS: Three (1.5 %) and 12 (6.2 %) were diagnosed as suffering from PCOS by Japanese and International criteria, respectively. 28.7 % of patients, (56/195) miscarried subsequently. There was no relation between diagnosis of PCOS, PCO morphology, elevated LH, free T, or obesity and subsequent miscarriage rate. CONCLUSIONS: A routine test for diagnosis of PCOS is not necessary in patients experiencing recurrent miscarriage because none of the related parameters examined in the present study predicted subsequent miscarriage. Supported by: Aichi Health Promotion Foundation.
P-12 THE ROLE OF MAJOR HISTOCOMPATIBILITY COMPLEX IN UNEXPLAINED RECURRENT PREGNANCY LOSS. H. Karagozoglu, S. Kahraman, M. O. Elbasi, E. Demiralp. ART Center, Istanbul Memorial Hospital, Istanbul, Turkey; Department of Hematology and Immunology, Marmara University, School of Medicine, Istanbul, Turkey. OBJECTIVE: Recurrent spontaneous abortions (RSA) are defined as 3 or more consecutive pregnancy losses under 20 weeks of gestation and present in 0.5-1% of fertile couples. The reason for RSA could not be identified in nearly half of the cases despite comprehensive studies and remained as unexplained. Maternal tolerance to the foetus during pregnancy is of great importance for the continuation of foetal life. Therefore, Major Histocompatibility Complex (MHC) may play a role in unexplained RSA. In this study we have aimed to investigate whether MHC similarities or differences between the partners are related to unexplained RSA. DESIGN: MHC tissue compatibilities and HLA-allele frequencies were examined in a selected group of patients with idiopathic RSA. MATERIALS AND METHODS: Twenty-four women diagnosed as RSA and their partners have been investigated for MHC class I and class II groups. 16 women with their partners who had successful deliveries before were also added as a control group. Genomic DNAs from patients and controls were isolated by using silica gel-based method. Low-resolution SSP typing was performed for identification of HLA-A,-B,-C and HLA-DR,-DQ groups. Chi-square test was used in comparisons. RESULTS: Examination of tissue group compatibilities in patients with RSA and controls revealed that, in all RSA patients, at least 1 MHC was compatible with their partners. Furthermore, 2 and 3 tissue groups were found to be compatible in 38.4% and 15.3% of the couples, respectively. On the other hand, in the control group, at least 1 tissue group was found to be compatible in 86.6% of the couples; 3 tissue groups were found to be compatible in 23% of the couples, 5 tissue groups were found to be compatible in 30.7% of the couples. HLA-Cw4 and HLA-Cw7 alleles were frequently detected in women (24% and 18.4%, respectively) and HLA-Cw7 was detected in 29.5 % of the men in the RSA group. While HLA-Cw7 allele was found in 14.2% of the women in the control group; HLA-Cw7 allele could not be detected in men. CONCLUSIONS: Initial data shows that non-identical allele frequency in the RSA group is statistically significantly higher than in the control group
Vol. 90, Suppl 1, September 2008
P-10 PREDICTION OF ABNORMAL PREGNANCY BY TROPHOBLAST ASSESSMENT IN FIRST TRIMESTER TRANSCERVICAL SAMPLES. A. N. Imudia, Y. Suzuki, F. D. Yelian, M. P. Diamond, R. Romero, D. R. Armant. Obstetrics and Gynecology, Wayne State University, Detroit, MI; Perinatology Research Branch, Bethesda, MD; Reproductive Biology & Medicine Branch, Bethesda, MD. OBJECTIVE: Ectopic pregnancy complicates about 1-2% of all pregnancies and occurs when the developing blastocyst implants at a site other than the fundus of the uterine cavity, most commonly in the fallopian tube. Delayed clinical diagnosis of this abnormality can result in a dismal maternal outcome. The presence of fetal trophoblast cells in the cervical canal during the first trimester suggested a non-invasive approach for predicting abnormal pregnancy through transcervical sampling. DESIGN: Experimental pilot study. MATERIALS AND METHODS: This study was approved by the Wayne State University IRB and written informed consent was obtained from participating women. Patients, age 18-40, presenting for normal early prenatal care (n¼10), ectopic pregnancy (EP; n¼7), or blighted ovum (BO; n¼5) were enrolled for collection of transcervical specimens using a cytological brush and a ThinPrepÒ kit (Hologic). Cells collected in PreservCytÒ fixative solution were cleared of mucus by acidification, washed by centrifugation and an aliquot was prepared on a microscope slide using a Cytospin3 centrifuge. Slides were labeled with monoclonal antibody G233 recognizing HLA-G, a MHC antigen specifically expressed by trophoblast cells. All HLA-G positive cells were identified and counted on each slide. After staining with hematoxylin, the total number of cells on each slide was determined and the ratio of HLA-G positive cells to total cells was calculated. Data were compared using one-way ANOVA, the Student-Newman-Keuls posthoc test and receiver operating characteristic analysis (ROC). RESULTS: The mean gestational ages of normal intrauterine pregnancy (IUP), EP, and BO were 9, 8, and 10 weeks, respectively. Trophoblast cells were observed in 10/10 normal IUP, 3/7 EP, and 4/5 BO specimens. The frequency of HLA-G positive cells in the IUP cervical samples was approximately 1 in 2000, which was 5- to 10-fold higher (p<0.001) than the average frequency in samples from patients with EP or BO. Significantly, the identification of trophoblast cells in cervical samples demonstrated both 100% sensitivity and 100% specificity. CONCLUSIONS: Based on limited observations, trophoblast cells can be reliably identified among transcervical cells in the first trimester by immunohistochemical staining for HLA-G. This non-invasive procedure may be useful for prenatal diagnosis of pregnancy outcomes and could be adapted for genetic analysis, but will require further clinical investigation to establish its efficacy. Supported by: The intramural research program of NICHD.
S114
Abstracts
P-11 THE POLYCYSTIC OVARY SYNDROME IN JAPANESE RECURRENT MISCARRIAGE PATIENTS. M. Sugiura-Ogasawara, Y. Ozaki, T. Nakanishi, K. Nozawa, S. Obayashi. Obstetrics and Gynecology, Nagoya City University Medical School, Nagoya, Aichi, Japan; Nagoya City University Medical School, Nagoya, Japan. OBJECTIVE: The polycystic ovary syndrome (PCOS) is well-known to be associated with miscarriages. However, it has been a matter of controversy whether it is actually a causal factor because of the absence of internationally established criteria. We therefore here investigated whether PCOS and a polycystic ovary (PCO) morphology have predictive value for subsequent miscarriages using new International and Japanese criteria. DESIGN: Prospective study. MATERIALS AND METHODS: A total of 195 patients with a history of two consecutive first trimester miscarriages and no abnormal chromosomes in either partner, antiphospholipid antibodies or uterine anomalies, were examined. The prospective pregnancy outcome was compared between patients with and without PCOS, PCO morphology, elevated LH, hyperandrogenism, and obesity. RESULTS: Three (1.5 %) and 12 (6.2 %) were diagnosed as suffering from PCOS by Japanese and International criteria, respectively. 28.7 % of patients, (56/195) miscarried subsequently. There was no relation between diagnosis of PCOS, PCO morphology, elevated LH, free T, or obesity and subsequent miscarriage rate. CONCLUSIONS: A routine test for diagnosis of PCOS is not necessary in patients experiencing recurrent miscarriage because none of the related parameters examined in the present study predicted subsequent miscarriage. Supported by: Aichi Health Promotion Foundation.
P-12 THE ROLE OF MAJOR HISTOCOMPATIBILITY COMPLEX IN UNEXPLAINED RECURRENT PREGNANCY LOSS. H. Karagozoglu, S. Kahraman, M. O. Elbasi, E. Demiralp. ART Center, Istanbul Memorial Hospital, Istanbul, Turkey; Department of Hematology and Immunology, Marmara University, School of Medicine, Istanbul, Turkey. OBJECTIVE: Recurrent spontaneous abortions (RSA) are defined as 3 or more consecutive pregnancy losses under 20 weeks of gestation and present in 0.5-1% of fertile couples. The reason for RSA could not be identified in nearly half of the cases despite comprehensive studies and remained as unexplained. Maternal tolerance to the foetus during pregnancy is of great importance for the continuation of foetal life. Therefore, Major Histocompatibility Complex (MHC) may play a role in unexplained RSA. In this study we have aimed to investigate whether MHC similarities or differences between the partners are related to unexplained RSA. DESIGN: MHC tissue compatibilities and HLA-allele frequencies were examined in a selected group of patients with idiopathic RSA. MATERIALS AND METHODS: Twenty-four women diagnosed as RSA and their partners have been investigated for MHC class I and class II groups. 16 women with their partners who had successful deliveries before were also added as a control group. Genomic DNAs from patients and controls were isolated by using silica gel-based method. Low-resolution SSP typing was performed for identification of HLA-A,-B,-C and HLA-DR,-DQ groups. Chi-square test was used in comparisons. RESULTS: Examination of tissue group compatibilities in patients with RSA and controls revealed that, in all RSA patients, at least 1 MHC was compatible with their partners. Furthermore, 2 and 3 tissue groups were found to be compatible in 38.4% and 15.3% of the couples, respectively. On the other hand, in the control group, at least 1 tissue group was found to be compatible in 86.6% of the couples; 3 tissue groups were found to be compatible in 23% of the couples, 5 tissue groups were found to be compatible in 30.7% of the couples. HLA-Cw4 and HLA-Cw7 alleles were frequently detected in women (24% and 18.4%, respectively) and HLA-Cw7 was detected in 29.5 % of the men in the RSA group. While HLA-Cw7 allele was found in 14.2% of the women in the control group; HLA-Cw7 allele could not be detected in men. CONCLUSIONS: Initial data shows that non-identical allele frequency in the RSA group is statistically significantly higher than in the control group
Vol. 90, Suppl 1, September 2008