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Assisted reproduction in histopathologically proven endometrial kochs
THURSDAY,
SEPTEMBER
7
FC4.13.02 KINETICS OF IGFBP-1 IN CERVIX DURING MEDICAL TERMINATION OF EARLY HUMAN PREGNANCY H. Ho&men, E.-M. Rutanen, 0. Heikinheimo, Dept. OBIGYN, University Hospital, Helsinki, Finland Objectives: To study the kinetics of decidual and amniotic fluid isoforms of insulin-like growth factor binding protein-l (IGFBP-1) in cervical secretion during abortion process induced by mifepristone and misoprostol. Study methods: The concentrations of IGFBP-1 were measured in cervical swab samples of 24 patients undergoing medical termination of early pregnancy with 200 mg of mifepristone orally followed by 0.8 mg of misoprostol orally or vaginally two days later. The samples were collected on day 0 (before mifepristone), twice on day 2 (before and 3 hours after misoprostol), day 14 and day 42. Two immunoenzymometric assays were used. Assay 1 detects the nonphosphorylated and lesser phosphorylated isoforms, those present in amniotic fluid (AF). Assay 2 detects the highly phosphorylated isoform of IGFBP-1, which is the primary isoform in decidua but is not present in AF. Results: IGFBP-1 concentrations in cervical samples started to rise after intake of mifepristone to reach the peak value after administration of misoprostol. The median values by assay 1 were 3 (day 0), 40 (day 2), 560 (day 2 +3 hours) -g/l. By assay 2 they were: 6 (day 0), 73 (day 2), 200 (day 2 + 3 hours) s/l. By day 14, the concentrations of IGFBP-1 had declined to basal levels. Conclusions: In intact early pregnancy, decidual isoforms of IGFBP-1 predominate in cervical secretion. Slight decidual bleeding after intake of mifepristone results in an increase of cervical IGFBP-1 concentration. Rupture of fetal membranes, which is likely to occur after administration of misoprostol, is switching the ratio of deciduaVamniotic fluid isoforms to opposite.
FC4.13.03 CONGENITAL MALFORMATION OCCURRING AS A RESULT OF PRE-IMPLANTATION EXPOSURE OF MOUSE EMBRYOS TO AMMONIUM CHLORIDE S. Dept. OBIGYN, Faculty of Medicine, Khon Kaen University, Amphur Maung, Thailand. Objectives: To investigate the effects of pre-implantation exposure to ammonium chloride on the development of (FlxFl) strain mouse embryos. Study Methods: A total of 280, two-cell stage mouse embryos were randomly allocated to culture in either Ml6 medium or Ml6 added with 0.3mM ammonium chloride for 2 days before being transferred to 2.5 day pseudopregnant recipients. Embryo morphology was assessed after 1 and 2 days of culture. The recipient females were sacrificed on day 15.5 of gestation. The number of implantation sites, fetuses, moles and any gross abnormality found were noted. Results: Pre-implantation exposure to ammonium chloride gave rise to a significant increase in the number of embryos reaching the morula stage after two days of culture ($=8.93, P
45
FC4.13.04 ULTRASTRUCTURAL IMAGING OF HUMAN IN VITRO FERTILIZATION AND EARLY EMBRYOGENESIS S. Makabe (l), T.Naguro (2), P.M. Motta (3) (1) Dept. OBIGYN, University of Toho, Tokyo, Japan. (2) Dept. of Anatomy, Faculty of Medicine, Tottori University, Tottori, Japan. (3) Dept. of Anatomy, Faculty of Medicine, University of Rome “La Sapienza”, Rome, Italy. Objectives: The aim of the study was to investigate ultrastructural morphodynamic events in human in vitro fertilization (IVF) by transmission (TEM) and field emission scanning electron microscopy (SEM). Study Methods: The materials (obtained with patients informed consent) included early human embryos and blastocysts after conventional IVF or intracytoplasmic sperm injection (ICSI). In addition ten cases of spontaneous parthenogenesis (2.cell stage) were also observed for the first time. The samples were first frozen/cracked and studied by SEM following the ODO maceration method (1,2). The specimens were observed with a SEM (Hitachi S-4500) and a TEM Zeiss MlO. Results: Great amounts of smooth endoplasmic reticulum (SER) aggregates were found in 4,5 and 6-cell embryos after ICSI. On the contrary, such structures were never found in embryos at the same developmental stages obtained after conventional IVF. Furthermore the inside overview of the three-dimensional (3-D) blastocysts was shown. Spontaneous parthenogenesis (2.cell) stage revealed a number of coritcal granules still assembled in the cytoplasmic cortex of parthenogenetic eggs. Conclusions: The Large SER aggregates often present in the blastomores of early embryos (4-6 cell stage) following ICSI procedures were peculiar structures never detected so far in similar studies in humans. Their significance remains unknown and highly speculative. The above techniques represent a useful and valid guideline to compare and correlate more exactly physiological and pathological events (3) of clinical relevance in assisted reproduction. References 1) Tanaka K. and Naguro T., Biomed Res. 2:63-70,1981; 2) Makabe S et al., Arch. Hist. Cytol. 57:389-394, 1994; 3) Motta P.M. et al, Protoplasma 206:270-77, 1999.
FC4.13.05 ASSISTED REPRODUCTION IN HISTOPATHOLOGICALLY PROVEN ENDOMETRIAL KOCHS S. Desai, P.G. Roy, M.D. Hansotia, Dept. OBIGYN, Fertility Clinic & IVF Center, Mumbai, Maharashtra, India. Objectives: Genital tuberculosis causes devastating effects on the reproductive functions of the woman. It causes irreparably damaged fallopian tubes. The problem mounts when the endometrium is also affected. We present our experience with infertile women affected with histopathologically proven endometrial tuberculosis who underwent ART at our Center, Fertility Clinic, Mumbai, India. Methods: 42 patients in the age group of 25-40 years underwent ART after proper evaluation including hysteroscopy to exclude intrauterine adhesions. All these women had received prior multidrug chemotherapy for TB endometritis and were free from the disease prior to ART. All but one were cases of primary infertility. 35 women had irreparably blocked tubes, 12 had frozen pelvis, 11 had bilateral T-O masses and one underwent unilateral salpingo oophorectomy for TB abscess. 5 patients underwent hysteroscopic adhesiolysis prior to IVF. Cycle monitoring also included measurement of endometrial thickness and color doppler study of the endometrium. Results: In 39 patients, 53 ovum pickups were done. 3 patients were cancelled due to poor ovarian response. There were 11 clinical pregnancies. Pregnancy rate was 2O%/OPU and 28%/Patient. No pregnancies were recorded in those who had endometrial thickness of 8 mm or less at the time of HCG injection. 4 women in the age group of 24-29 years responded poorly, possibly due to diseased ovaries. Conclusion: This series represents a comparatively encouraging success rate in these carefully selected patients of tuberculous endometritis. ART offers hope to this problematic group.
SEPTEMBER
7
FC4.13.02 KINETICS OF IGFBP-1 IN CERVIX DURING MEDICAL TERMINATION OF EARLY HUMAN PREGNANCY H. Ho&men, E.-M. Rutanen, 0. Heikinheimo, Dept. OBIGYN, University Hospital, Helsinki, Finland Objectives: To study the kinetics of decidual and amniotic fluid isoforms of insulin-like growth factor binding protein-l (IGFBP-1) in cervical secretion during abortion process induced by mifepristone and misoprostol. Study methods: The concentrations of IGFBP-1 were measured in cervical swab samples of 24 patients undergoing medical termination of early pregnancy with 200 mg of mifepristone orally followed by 0.8 mg of misoprostol orally or vaginally two days later. The samples were collected on day 0 (before mifepristone), twice on day 2 (before and 3 hours after misoprostol), day 14 and day 42. Two immunoenzymometric assays were used. Assay 1 detects the nonphosphorylated and lesser phosphorylated isoforms, those present in amniotic fluid (AF). Assay 2 detects the highly phosphorylated isoform of IGFBP-1, which is the primary isoform in decidua but is not present in AF. Results: IGFBP-1 concentrations in cervical samples started to rise after intake of mifepristone to reach the peak value after administration of misoprostol. The median values by assay 1 were 3 (day 0), 40 (day 2), 560 (day 2 +3 hours) -g/l. By assay 2 they were: 6 (day 0), 73 (day 2), 200 (day 2 + 3 hours) s/l. By day 14, the concentrations of IGFBP-1 had declined to basal levels. Conclusions: In intact early pregnancy, decidual isoforms of IGFBP-1 predominate in cervical secretion. Slight decidual bleeding after intake of mifepristone results in an increase of cervical IGFBP-1 concentration. Rupture of fetal membranes, which is likely to occur after administration of misoprostol, is switching the ratio of deciduaVamniotic fluid isoforms to opposite.
FC4.13.03 CONGENITAL MALFORMATION OCCURRING AS A RESULT OF PRE-IMPLANTATION EXPOSURE OF MOUSE EMBRYOS TO AMMONIUM CHLORIDE S. Dept. OBIGYN, Faculty of Medicine, Khon Kaen University, Amphur Maung, Thailand. Objectives: To investigate the effects of pre-implantation exposure to ammonium chloride on the development of (FlxFl) strain mouse embryos. Study Methods: A total of 280, two-cell stage mouse embryos were randomly allocated to culture in either Ml6 medium or Ml6 added with 0.3mM ammonium chloride for 2 days before being transferred to 2.5 day pseudopregnant recipients. Embryo morphology was assessed after 1 and 2 days of culture. The recipient females were sacrificed on day 15.5 of gestation. The number of implantation sites, fetuses, moles and any gross abnormality found were noted. Results: Pre-implantation exposure to ammonium chloride gave rise to a significant increase in the number of embryos reaching the morula stage after two days of culture ($=8.93, P
45
FC4.13.04 ULTRASTRUCTURAL IMAGING OF HUMAN IN VITRO FERTILIZATION AND EARLY EMBRYOGENESIS S. Makabe (l), T.Naguro (2), P.M. Motta (3) (1) Dept. OBIGYN, University of Toho, Tokyo, Japan. (2) Dept. of Anatomy, Faculty of Medicine, Tottori University, Tottori, Japan. (3) Dept. of Anatomy, Faculty of Medicine, University of Rome “La Sapienza”, Rome, Italy. Objectives: The aim of the study was to investigate ultrastructural morphodynamic events in human in vitro fertilization (IVF) by transmission (TEM) and field emission scanning electron microscopy (SEM). Study Methods: The materials (obtained with patients informed consent) included early human embryos and blastocysts after conventional IVF or intracytoplasmic sperm injection (ICSI). In addition ten cases of spontaneous parthenogenesis (2.cell stage) were also observed for the first time. The samples were first frozen/cracked and studied by SEM following the ODO maceration method (1,2). The specimens were observed with a SEM (Hitachi S-4500) and a TEM Zeiss MlO. Results: Great amounts of smooth endoplasmic reticulum (SER) aggregates were found in 4,5 and 6-cell embryos after ICSI. On the contrary, such structures were never found in embryos at the same developmental stages obtained after conventional IVF. Furthermore the inside overview of the three-dimensional (3-D) blastocysts was shown. Spontaneous parthenogenesis (2.cell) stage revealed a number of coritcal granules still assembled in the cytoplasmic cortex of parthenogenetic eggs. Conclusions: The Large SER aggregates often present in the blastomores of early embryos (4-6 cell stage) following ICSI procedures were peculiar structures never detected so far in similar studies in humans. Their significance remains unknown and highly speculative. The above techniques represent a useful and valid guideline to compare and correlate more exactly physiological and pathological events (3) of clinical relevance in assisted reproduction. References 1) Tanaka K. and Naguro T., Biomed Res. 2:63-70,1981; 2) Makabe S et al., Arch. Hist. Cytol. 57:389-394, 1994; 3) Motta P.M. et al, Protoplasma 206:270-77, 1999.
FC4.13.05 ASSISTED REPRODUCTION IN HISTOPATHOLOGICALLY PROVEN ENDOMETRIAL KOCHS S. Desai, P.G. Roy, M.D. Hansotia, Dept. OBIGYN, Fertility Clinic & IVF Center, Mumbai, Maharashtra, India. Objectives: Genital tuberculosis causes devastating effects on the reproductive functions of the woman. It causes irreparably damaged fallopian tubes. The problem mounts when the endometrium is also affected. We present our experience with infertile women affected with histopathologically proven endometrial tuberculosis who underwent ART at our Center, Fertility Clinic, Mumbai, India. Methods: 42 patients in the age group of 25-40 years underwent ART after proper evaluation including hysteroscopy to exclude intrauterine adhesions. All these women had received prior multidrug chemotherapy for TB endometritis and were free from the disease prior to ART. All but one were cases of primary infertility. 35 women had irreparably blocked tubes, 12 had frozen pelvis, 11 had bilateral T-O masses and one underwent unilateral salpingo oophorectomy for TB abscess. 5 patients underwent hysteroscopic adhesiolysis prior to IVF. Cycle monitoring also included measurement of endometrial thickness and color doppler study of the endometrium. Results: In 39 patients, 53 ovum pickups were done. 3 patients were cancelled due to poor ovarian response. There were 11 clinical pregnancies. Pregnancy rate was 2O%/OPU and 28%/Patient. No pregnancies were recorded in those who had endometrial thickness of 8 mm or less at the time of HCG injection. 4 women in the age group of 24-29 years responded poorly, possibly due to diseased ovaries. Conclusion: This series represents a comparatively encouraging success rate in these carefully selected patients of tuberculous endometritis. ART offers hope to this problematic group.