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Regulation of expression of p57KIP2 gene in extravillous trophoblast cells
A20
Abstracts / Placenta 35 (2014) A1eA23
mesenchymal hamartoma. To perform chromosome analysis also hamartoma similarly, as a result, we have confirmed the presence of 5th trisomy. It is expected the possibility that ABM is associated with fetal abnormality can be considered advances the scrutiny also other organs. Further investigation of additional mosaic case may elucidate the mechanisms and phenotypes.
O-081. REGULATION OF EXPRESSION OF P57KIP2 GENE IN EXTRAVILLOUS TROPHOBLAST CELLS Hirokazu Usui, Jia Qu, Emiri Nakata, Hiroshi Kaku, Shinsuke Hanawa, Tatsuya Kobayashi, Makio Shozu. Department of Reproductive Medicine, Chiba University Graduate School of Medicine, Japan Introduction: The p57KIP2 gene is located in the imprinted gene cluster on human chromosome 11, and works inhibitory to cell growth. The immunostaining of p57KIP2 is negative in androgenetic complete hydatidiform moles (CHM) because they do not have maternal genome. It is useful to differentiate molar pregnancies in histological diagnosis. In extravillous trophoblast in CHM, the immunostaining of p57KIP2 is positive although they do not have any maternal genome. The mechanism of p57KIP2 expression in EVT of CHM is unveiled. We hypothesized that there is some machinery that excessive growth of EVT is inhibited by elevation of p57KIP2 expression. To clarify this hypothesis, we analyzed the expression and regulatory mechanism of p57KIP2 gene in EVT between CHM and biparental diploid villous tissue (D-villi). Methods: EVTs were purified by the primary cultures from CHM and Dvilli, confirmed genetically by short tandem repeat polymorphism PCR. 1) Real-time PCR was performed for the expression of p57KIP2 gene in EVT and villous tissue. 2) We evaluated the methylation status of KvDMR1 region that epigenetically regulate the p57KIP2 gene by MLPA and COBRA. 3) The mRNA origin was confirmed by allele-specific RT-PCR with SNPs and direct sequencing. Results: 1) The expressions of p57KIP2 in EVTs of CHM and D-villi were higher than those of villous tissue. 2) Methylation status of KvDMR1 did not change between EVT and villous tissue. 3) Only maternal origin of mRNA was translated in both EVT and villous tissue in D-villi. Various forms of splicing variant of p57KIP2 in exon 2 were identified for the first time. Conclusion: The expression of p57KIP2 gene was elevated in EVT, although the methylation status of them did not alter. The regulation of p57KIP2 in EVT of CHM may be related to the transcriptional level of mechanism.
O-085. HOW THE PLACENTA IN TRISOMIES 13, 18, AND 21 EXPRESS THEMSELVES.
such as occlusion, recanalization and calcification of vascular walls. Trisomy 13 features dysmature villi in both large and small sizes. The villi of trisomy 21 displayed a clear trophoblast layer through immune staining of HCG. Some trisomic placenta showed iron stain positive on the basement membrane. Consideration: 1. The villi of trisomy 21 have a very unique shape and number of vessels, amongst other things, and these characteristics make diagnosis very simple. 2. If TAM is discovered, it can be stated with certainty that trisomy 21 must be present since TAM consists of GATA1 and trisomy 21 chromosome. 3. The placentas of trisomy 18 have vascular lesions such as occlusion, recanalization and calcification of vascular walls. The baby also has other vascular problems. 4. The placentas of trisomy 13 are small due to dysmature villi and ischemic villi which causes trisomy 13 baby’s weight to be light. 5. Iron is too much for some trisomic fetus growth.
O-086. HISTOLOGIC EVIDENCES FOR CHORIOAMNIONITIS AND DECIDUAL VASCULAR LESION AS HIGH RISK CAUSES OF ABRUPTIO PLACENTAE Masayoshi Arizawa. Tokyo Metropolitan Ohtsuka Hospital Department of Laboratory Medicine, Japan Back ground: In abruption, the placenta detaches while the baby is still in the uterus, therefore the baby falls into hypoxia, and the mother has crisis of bleeding. The baby may suffer a variety of conditions or even be born dead. Purpose: It is said that the exact causes of abruptio placentae may be hard to determine clinically, however, in my work I have often come across significant indicators under microscope. Namely, high grade inflammation related to chorioamnionitis (CAM) and ischemic villi caused by decidual vascular lesion. In this article I show how abruptio placentae are relate to CAM and ischemic villi in the placenta. Material: I reexamined placentas of abruptio placentae pathologically, those with divided decidua, invasive bleeding and necrosis. Results: Out of 225 abruptio placentae cases I examined, 87 had CAM and 75 had ischemic change. Of the 87 CAM cases, 63 were before 32 gestational weeks, and 24 cases over 32 gestational weeks. Of 75 ischemic cases 33 were before 32 gestational weeks and 42 were over 32 gestational weeks. Conclusion: My observations suggest that the association between CAM and abruptio placentae is strongest before 32 gestational weeks. So it is important to be aware of the risks of infection, because of the possibility of abruptio placentae. Ischemic villi is also a great cause of abruptio placentae across all gestational weeks, This is due to PIH caused by mother’s vascular problems (Poor trophoblast migration hinders efficient dilatation), and baby’s hypoxia (Adding increase demand upon mother’s blood flow, increased mother’s pregnancy induced hypertension (PIH)), which we must be aware of at all stages.
Masayoshi Arizawa. Tokyo Metropolitan Ohtsuka Hospital Department of laboratory Medicine, Japan Purpose: One paper explained that trisomy 21 is heavier than other trisomies.48 placental examinations of the trisomies 13, 18, and 21 under the microscope, showed the dysmature villi of trisomy 21 had a large number of blood vessels, and dysmature villi of the trisomies 18 and 13 showed smaller amounts of blood vessels. This time I have revisited this previous work, with new placenta and examinations. Method: I selected placental specimens of trisomies 13, 18, and 21 from placental forms and medical records. I used HCG immune staining on trisomy 21 placentas, and I used iron staining on trisomy 13, 18, and 21 placentas. Results: I discovered the following cases: 6 cases of trisomy 13, 8 cases of trisomy 18, 26 cases of trisomy 21; 40 cases in total. As a result of microscopic examination, I found that trisomy 21 features, large, dysmature villi, Chorangiosis was diagnosed in 14 of the 26 cases of trisomy 21. TAM (Transient abnormal myelopoiesis) was diagnosed in 6 of the 26 cases of trisomy 21. Trisomy 18 features dysmature villi with blood vessel disorders
O-087. CLINICAL PICTURES AND PATHOLOGICAL EXAMINATION MONOCHORIONIC-DIAMNIOTIC (MD) TWIN PREGNANCY
OF
Masayoshi Arizawa. Tokyo Metropolitan Ohtsuka Hospital Department of Laboratory Medicine, Japan Purpose: Monochorionic-Diamniotic (MD) twin pregnancy is a sub type of monochorionic twin pregnancy. These pregnancies share a single chorion. There are 2 major conditions of MD twin. 1. Placental umbilical cord insertion related problems. Increased incidence of velamentous cord insertion and marginal cord insertion. 2. Twin-to-twin transfusion syndrome (TTTS). A condition whereby a donor twin bleeds into the circulation of a recipient twin. The result is the recipient twin will grow larger, become polycytemia and will have excessive amniotic fluid (polyhydramnios), while the donor twin will experience growth retardation, become anemic and will have too little amniotic fluid (oligohydramnios).
Abstracts / Placenta 35 (2014) A1eA23
mesenchymal hamartoma. To perform chromosome analysis also hamartoma similarly, as a result, we have confirmed the presence of 5th trisomy. It is expected the possibility that ABM is associated with fetal abnormality can be considered advances the scrutiny also other organs. Further investigation of additional mosaic case may elucidate the mechanisms and phenotypes.
O-081. REGULATION OF EXPRESSION OF P57KIP2 GENE IN EXTRAVILLOUS TROPHOBLAST CELLS Hirokazu Usui, Jia Qu, Emiri Nakata, Hiroshi Kaku, Shinsuke Hanawa, Tatsuya Kobayashi, Makio Shozu. Department of Reproductive Medicine, Chiba University Graduate School of Medicine, Japan Introduction: The p57KIP2 gene is located in the imprinted gene cluster on human chromosome 11, and works inhibitory to cell growth. The immunostaining of p57KIP2 is negative in androgenetic complete hydatidiform moles (CHM) because they do not have maternal genome. It is useful to differentiate molar pregnancies in histological diagnosis. In extravillous trophoblast in CHM, the immunostaining of p57KIP2 is positive although they do not have any maternal genome. The mechanism of p57KIP2 expression in EVT of CHM is unveiled. We hypothesized that there is some machinery that excessive growth of EVT is inhibited by elevation of p57KIP2 expression. To clarify this hypothesis, we analyzed the expression and regulatory mechanism of p57KIP2 gene in EVT between CHM and biparental diploid villous tissue (D-villi). Methods: EVTs were purified by the primary cultures from CHM and Dvilli, confirmed genetically by short tandem repeat polymorphism PCR. 1) Real-time PCR was performed for the expression of p57KIP2 gene in EVT and villous tissue. 2) We evaluated the methylation status of KvDMR1 region that epigenetically regulate the p57KIP2 gene by MLPA and COBRA. 3) The mRNA origin was confirmed by allele-specific RT-PCR with SNPs and direct sequencing. Results: 1) The expressions of p57KIP2 in EVTs of CHM and D-villi were higher than those of villous tissue. 2) Methylation status of KvDMR1 did not change between EVT and villous tissue. 3) Only maternal origin of mRNA was translated in both EVT and villous tissue in D-villi. Various forms of splicing variant of p57KIP2 in exon 2 were identified for the first time. Conclusion: The expression of p57KIP2 gene was elevated in EVT, although the methylation status of them did not alter. The regulation of p57KIP2 in EVT of CHM may be related to the transcriptional level of mechanism.
O-085. HOW THE PLACENTA IN TRISOMIES 13, 18, AND 21 EXPRESS THEMSELVES.
such as occlusion, recanalization and calcification of vascular walls. Trisomy 13 features dysmature villi in both large and small sizes. The villi of trisomy 21 displayed a clear trophoblast layer through immune staining of HCG. Some trisomic placenta showed iron stain positive on the basement membrane. Consideration: 1. The villi of trisomy 21 have a very unique shape and number of vessels, amongst other things, and these characteristics make diagnosis very simple. 2. If TAM is discovered, it can be stated with certainty that trisomy 21 must be present since TAM consists of GATA1 and trisomy 21 chromosome. 3. The placentas of trisomy 18 have vascular lesions such as occlusion, recanalization and calcification of vascular walls. The baby also has other vascular problems. 4. The placentas of trisomy 13 are small due to dysmature villi and ischemic villi which causes trisomy 13 baby’s weight to be light. 5. Iron is too much for some trisomic fetus growth.
O-086. HISTOLOGIC EVIDENCES FOR CHORIOAMNIONITIS AND DECIDUAL VASCULAR LESION AS HIGH RISK CAUSES OF ABRUPTIO PLACENTAE Masayoshi Arizawa. Tokyo Metropolitan Ohtsuka Hospital Department of Laboratory Medicine, Japan Back ground: In abruption, the placenta detaches while the baby is still in the uterus, therefore the baby falls into hypoxia, and the mother has crisis of bleeding. The baby may suffer a variety of conditions or even be born dead. Purpose: It is said that the exact causes of abruptio placentae may be hard to determine clinically, however, in my work I have often come across significant indicators under microscope. Namely, high grade inflammation related to chorioamnionitis (CAM) and ischemic villi caused by decidual vascular lesion. In this article I show how abruptio placentae are relate to CAM and ischemic villi in the placenta. Material: I reexamined placentas of abruptio placentae pathologically, those with divided decidua, invasive bleeding and necrosis. Results: Out of 225 abruptio placentae cases I examined, 87 had CAM and 75 had ischemic change. Of the 87 CAM cases, 63 were before 32 gestational weeks, and 24 cases over 32 gestational weeks. Of 75 ischemic cases 33 were before 32 gestational weeks and 42 were over 32 gestational weeks. Conclusion: My observations suggest that the association between CAM and abruptio placentae is strongest before 32 gestational weeks. So it is important to be aware of the risks of infection, because of the possibility of abruptio placentae. Ischemic villi is also a great cause of abruptio placentae across all gestational weeks, This is due to PIH caused by mother’s vascular problems (Poor trophoblast migration hinders efficient dilatation), and baby’s hypoxia (Adding increase demand upon mother’s blood flow, increased mother’s pregnancy induced hypertension (PIH)), which we must be aware of at all stages.
Masayoshi Arizawa. Tokyo Metropolitan Ohtsuka Hospital Department of laboratory Medicine, Japan Purpose: One paper explained that trisomy 21 is heavier than other trisomies.48 placental examinations of the trisomies 13, 18, and 21 under the microscope, showed the dysmature villi of trisomy 21 had a large number of blood vessels, and dysmature villi of the trisomies 18 and 13 showed smaller amounts of blood vessels. This time I have revisited this previous work, with new placenta and examinations. Method: I selected placental specimens of trisomies 13, 18, and 21 from placental forms and medical records. I used HCG immune staining on trisomy 21 placentas, and I used iron staining on trisomy 13, 18, and 21 placentas. Results: I discovered the following cases: 6 cases of trisomy 13, 8 cases of trisomy 18, 26 cases of trisomy 21; 40 cases in total. As a result of microscopic examination, I found that trisomy 21 features, large, dysmature villi, Chorangiosis was diagnosed in 14 of the 26 cases of trisomy 21. TAM (Transient abnormal myelopoiesis) was diagnosed in 6 of the 26 cases of trisomy 21. Trisomy 18 features dysmature villi with blood vessel disorders
O-087. CLINICAL PICTURES AND PATHOLOGICAL EXAMINATION MONOCHORIONIC-DIAMNIOTIC (MD) TWIN PREGNANCY
OF
Masayoshi Arizawa. Tokyo Metropolitan Ohtsuka Hospital Department of Laboratory Medicine, Japan Purpose: Monochorionic-Diamniotic (MD) twin pregnancy is a sub type of monochorionic twin pregnancy. These pregnancies share a single chorion. There are 2 major conditions of MD twin. 1. Placental umbilical cord insertion related problems. Increased incidence of velamentous cord insertion and marginal cord insertion. 2. Twin-to-twin transfusion syndrome (TTTS). A condition whereby a donor twin bleeds into the circulation of a recipient twin. The result is the recipient twin will grow larger, become polycytemia and will have excessive amniotic fluid (polyhydramnios), while the donor twin will experience growth retardation, become anemic and will have too little amniotic fluid (oligohydramnios).