- Email: [email protected]
Dental infection of porphyromonas gingivalis induces preterm birth
A22
Abstracts / Placenta 35 (2014) A1eA23
risk of stillbirth at GW N was defined as a number of women with stillbirth at GW. N divided by number of women giving birth at GW N. Results: A total of 32 (29.4%) women experienced stillbirth at median GW of 31 (range, 24±38). Preterm birth (GW37) occurred in 52 (67.5%) of the 77 live born infants. Only 25 (22.9%) women had full-term (GW37) live-born infants. The prospective risk of stillbirth was 29.4% (32/109), 27.5% (25/91), 20.9% (14/67) and 13.0% (6/46) for women who reached GW 24, 28, 32, and 36, respectively. Conclusions: As women with PMD faces at an extraordinary higher risk of IUFD, early admission to the hospital and intensive monitoring of foetal status should be considered, although whether this policy improves outcome has not been validated.
O-097. RADIOACTIVE CESIUM AND POTASSIUM LEVELS IN HUMAN PLACENTA: AFTER THE FUKUSHIMA NUCLEAR POWER PLANT ACCIDENT Makoto Kawamur a, b, Hiroshi Anbe b, Fujimori Keiya a. a Fukushima Medical University, Department of Obstetrics and Gynecology, Japan; b Minamisoma City General Hospital, Department of Obstetrics and Gynecology, Japan Background: The Great East Japan earthquake (followed by large-scale tsunamis) struck Northeastern Pacific Coast of Japan on Mar 11, 2011. It caused TEPCO's Fukushima Daiichi Nuclear Power Plant (NPP) accident, accompanied by radioactive pollution in Fukushima prefecture. Minamisoma City General Hospital, just 23km away from the NPP, restarted the Obstetric services in April 2012. We examined the placental radioactive Cesium and Potassium levels. Methods: We measured the radioactive contents of the 40samples of placenta after the delivery, using high-purity Germanium detectors for Gamma-ray spectrometry (at National Institute of Public Health, Wako, Saitama). All the parturients underwent the Whole-Body Counting (WBC) test of radioactivity. Results: The radioactive content levels are as follows. 134Cs: 0.36Bq/kg, 137 Cs: 0.58Bq/kg, total-Cs: 0.94Bq/kg, 40K (natural radioactive): 53.1Bq/kg. Radioactive Cs (from the NPP accident) levels were about 1/50 of the natural radioactive, and much lower than the '87 Italian data, 2years after the Chernobyl NPP accident (total-Cs:11.5Bq/kg). All of the parturients were negative on the WBC test (with cut-off value of 250Bq/body). Conclusion: The Cs levels were much lower than natural radioactive (40K) level, which suggests the efforts to reduce external/internal radioactive exposure in the area has been successful.
O-102. DENTAL INFECTION PRETERM BIRTH
OF
PORPHYROMONAS
GINGIVALIS
INDUCES
Mutsumi Miyauchi a, Hisako Furusho a, Atsuhiro Nagasaki a, Satoshi Urabe b, Haruhisa Konishi b, Hiroshi Miyoshi b, Yoshiki Kudo b, Takashi Takata a. a Department of Oral and Maxillofacial Pathobiology, Institute of Biomedical and Health Sciences, Hiroshima University, Japan; b Department of Obstetrics and Gynecology, Hiroshima University, Japan Objectives: Epidemiological studies revealed a link between periodontal disease and preterm delivery, however, the mechanism of the link remains unclear. The present study was aimed to examine the influence of Porphyromonas gingivalis (Pg): one of main periodontal pathogens) on preterm birth with a Pg dental infection mouse model and in vitro analyses with trophoblasts and related cells Methods: In vivo experiment: 8-wksold\C57Bl/6J mice were infected with Pg-w83-strain (108 CFU) from the pulp chamber (Pg group). Mating was started at 6-wks post-infection when a periapical granuloma was established. Gestational day (gd) and birth weight were examined.; At gd15, placental tissues were harvested for IHC staining and PCR. Serum, placental tissue and amniotic fluid were collected for ELISA of cytokines. Uninfected mice were used as negative control (NC). In vitro experiment: HTR-8 (trophoblast), HuhT1 (endothelial cell) and THP1 (monocyte) cell lines were used to assess the effects of Pg and Pg-LPS.
Results: Pg group showed 2 days of preterm birth (p<0.01) and low birth weight (p<0.01). The amnion-epithelium was damaged in placenta of Pg group and trophoblasts and endothelial cells became degenerative. Moreover, galectin-3 (Gal-3: an immune regulator) suffused the whole placental tissue and dramatically increased in amniotic fluid (P<0.01). The weaker immuno-expression of CD-31 indicated endothelial damage and immuno-expression of COX-2 and TNF-alpha increased in placental tissue of Pg group. Pg was detected in placental tissue. Pg-LPS up-regulated the expression of COX-2, TNF-alpha and Gal-3. Moreover, recombinant Gal-3 stimulation further increased expressions of them. Conclusion: Dentally infected Pg reaches placental tissue through blood circulation and induces preterm birth via upregulation of COX-2, TNF-a and Gal-3 expression in placental tissue.
O-103. EXPRESSION OF TOLL-LIKE RECEPTORS AND PROINFLAMMATORY CYTOKINES IN HUMAN PLACENTA Hiroaki Moroi, Tomomi Kotani, Yukio Mano, Hiroyuki Tsuda, Seiji Sumigama, Fumitaka Kikkawa. Department of Obstetrics and Gynecology, Nagoya University Graduate School of Medicine, Japan Objectives: Toll-like receptors (TLRs) are found at many sites in human body. They play important roles of the innate immune system. On the other hands, chorioamnionitis (CAM) is thought to be a cause of preterm birth (PTB) and subsequent neonatal adverse effect. It is well known that administration of lipopolysaccharide (LPS) which is recognized by TLR4 leads to preterm birth in animal model, and it supports that TLR4 is associated with CAM and PTB. However, there is little data about other members of TLRs. In this study, we focused on other TLR members and undertook investigation of the expression of these in the human preterm placenta to reveal the connection between TLRs and CAM. Methods: We extracted the 15 cases of PTB in our hospital. Immunohistochemistry was performed with each TLR antigen on microscopic section of the each placental tissues. Furthermore, we preformed immnofluorescence staining with TLR and IL6/8 antigen in the slides which were observed strongly positive staining of TLRs. On the other hand, we corrected human fetal membranes from the patients who received cesarean section without any perinatal complications. Tissues were cultured and stimulated by each TLR ligands, and the production of proinflamｍatory cytokines are evaluated by ELISA. Results: Immunohistochemistry showed increased expression of TLR4 and 5 in placental tissue of CAM (+) samples. But TLR 2 or 6 didn’t show significant difference between CAM(+) and CAM(-) groups. IL6/8 also strongly expressed in the same site in which TLR5 was stained. In the tissue culture, IL6 expression was remarkably increased by TLR2/4/5 ligands. But no significant difference of IL8 was seen. Conclusions: This study shows that, in chorioamnionitis, TLRs expression are elevated in the fetal membrane, and it may provide increased proinflammatory cytokine production. Therefore TLRs may pray important roles in human chorioamnionitis.
O-104. DNA METHYLATION ANALYSIS OF A CPG-RICH PROMOTER REGION OF C19MC IN HUMAN TROPHOBLAST CELLS Hironori Takahashi a, b, Akihide Ohkuchi a, Tomoyuki Kuwata a, Rie Usui a, Kunio Kikuchi b, Shigeki Matsubara a, Mitsuaki Suzuki a, Toshiyuki Takeshita c, Toshihiro Takizawa b. a Department of Obstetrics and Gynecology, Jichi Medical University, Tochigi, Japan; b Department of Molecular Medicine and Anatomy, Nippon Medical School, Tokyo, Japan; c Department of Reproductive Medicine, Perinatology and Gynecologic Oncology, Nippon Medical School, Tokyo, Japan Objective: In the human, miRNAs derived from the miRNA cluster on human chromosome 19 (C19MC), a primate-specific miRNA cluster, have been demonstrated to exhibit a placenta-specific expression pattern. The C19MC is an imprinting gene and is exclusively expressed from a paternal
Abstracts / Placenta 35 (2014) A1eA23
risk of stillbirth at GW N was defined as a number of women with stillbirth at GW. N divided by number of women giving birth at GW N. Results: A total of 32 (29.4%) women experienced stillbirth at median GW of 31 (range, 24±38). Preterm birth (GW37) occurred in 52 (67.5%) of the 77 live born infants. Only 25 (22.9%) women had full-term (GW37) live-born infants. The prospective risk of stillbirth was 29.4% (32/109), 27.5% (25/91), 20.9% (14/67) and 13.0% (6/46) for women who reached GW 24, 28, 32, and 36, respectively. Conclusions: As women with PMD faces at an extraordinary higher risk of IUFD, early admission to the hospital and intensive monitoring of foetal status should be considered, although whether this policy improves outcome has not been validated.
O-097. RADIOACTIVE CESIUM AND POTASSIUM LEVELS IN HUMAN PLACENTA: AFTER THE FUKUSHIMA NUCLEAR POWER PLANT ACCIDENT Makoto Kawamur a, b, Hiroshi Anbe b, Fujimori Keiya a. a Fukushima Medical University, Department of Obstetrics and Gynecology, Japan; b Minamisoma City General Hospital, Department of Obstetrics and Gynecology, Japan Background: The Great East Japan earthquake (followed by large-scale tsunamis) struck Northeastern Pacific Coast of Japan on Mar 11, 2011. It caused TEPCO's Fukushima Daiichi Nuclear Power Plant (NPP) accident, accompanied by radioactive pollution in Fukushima prefecture. Minamisoma City General Hospital, just 23km away from the NPP, restarted the Obstetric services in April 2012. We examined the placental radioactive Cesium and Potassium levels. Methods: We measured the radioactive contents of the 40samples of placenta after the delivery, using high-purity Germanium detectors for Gamma-ray spectrometry (at National Institute of Public Health, Wako, Saitama). All the parturients underwent the Whole-Body Counting (WBC) test of radioactivity. Results: The radioactive content levels are as follows. 134Cs: 0.36Bq/kg, 137 Cs: 0.58Bq/kg, total-Cs: 0.94Bq/kg, 40K (natural radioactive): 53.1Bq/kg. Radioactive Cs (from the NPP accident) levels were about 1/50 of the natural radioactive, and much lower than the '87 Italian data, 2years after the Chernobyl NPP accident (total-Cs:11.5Bq/kg). All of the parturients were negative on the WBC test (with cut-off value of 250Bq/body). Conclusion: The Cs levels were much lower than natural radioactive (40K) level, which suggests the efforts to reduce external/internal radioactive exposure in the area has been successful.
O-102. DENTAL INFECTION PRETERM BIRTH
OF
PORPHYROMONAS
GINGIVALIS
INDUCES
Mutsumi Miyauchi a, Hisako Furusho a, Atsuhiro Nagasaki a, Satoshi Urabe b, Haruhisa Konishi b, Hiroshi Miyoshi b, Yoshiki Kudo b, Takashi Takata a. a Department of Oral and Maxillofacial Pathobiology, Institute of Biomedical and Health Sciences, Hiroshima University, Japan; b Department of Obstetrics and Gynecology, Hiroshima University, Japan Objectives: Epidemiological studies revealed a link between periodontal disease and preterm delivery, however, the mechanism of the link remains unclear. The present study was aimed to examine the influence of Porphyromonas gingivalis (Pg): one of main periodontal pathogens) on preterm birth with a Pg dental infection mouse model and in vitro analyses with trophoblasts and related cells Methods: In vivo experiment: 8-wksold\C57Bl/6J mice were infected with Pg-w83-strain (108 CFU) from the pulp chamber (Pg group). Mating was started at 6-wks post-infection when a periapical granuloma was established. Gestational day (gd) and birth weight were examined.; At gd15, placental tissues were harvested for IHC staining and PCR. Serum, placental tissue and amniotic fluid were collected for ELISA of cytokines. Uninfected mice were used as negative control (NC). In vitro experiment: HTR-8 (trophoblast), HuhT1 (endothelial cell) and THP1 (monocyte) cell lines were used to assess the effects of Pg and Pg-LPS.
Results: Pg group showed 2 days of preterm birth (p<0.01) and low birth weight (p<0.01). The amnion-epithelium was damaged in placenta of Pg group and trophoblasts and endothelial cells became degenerative. Moreover, galectin-3 (Gal-3: an immune regulator) suffused the whole placental tissue and dramatically increased in amniotic fluid (P<0.01). The weaker immuno-expression of CD-31 indicated endothelial damage and immuno-expression of COX-2 and TNF-alpha increased in placental tissue of Pg group. Pg was detected in placental tissue. Pg-LPS up-regulated the expression of COX-2, TNF-alpha and Gal-3. Moreover, recombinant Gal-3 stimulation further increased expressions of them. Conclusion: Dentally infected Pg reaches placental tissue through blood circulation and induces preterm birth via upregulation of COX-2, TNF-a and Gal-3 expression in placental tissue.
O-103. EXPRESSION OF TOLL-LIKE RECEPTORS AND PROINFLAMMATORY CYTOKINES IN HUMAN PLACENTA Hiroaki Moroi, Tomomi Kotani, Yukio Mano, Hiroyuki Tsuda, Seiji Sumigama, Fumitaka Kikkawa. Department of Obstetrics and Gynecology, Nagoya University Graduate School of Medicine, Japan Objectives: Toll-like receptors (TLRs) are found at many sites in human body. They play important roles of the innate immune system. On the other hands, chorioamnionitis (CAM) is thought to be a cause of preterm birth (PTB) and subsequent neonatal adverse effect. It is well known that administration of lipopolysaccharide (LPS) which is recognized by TLR4 leads to preterm birth in animal model, and it supports that TLR4 is associated with CAM and PTB. However, there is little data about other members of TLRs. In this study, we focused on other TLR members and undertook investigation of the expression of these in the human preterm placenta to reveal the connection between TLRs and CAM. Methods: We extracted the 15 cases of PTB in our hospital. Immunohistochemistry was performed with each TLR antigen on microscopic section of the each placental tissues. Furthermore, we preformed immnofluorescence staining with TLR and IL6/8 antigen in the slides which were observed strongly positive staining of TLRs. On the other hand, we corrected human fetal membranes from the patients who received cesarean section without any perinatal complications. Tissues were cultured and stimulated by each TLR ligands, and the production of proinflamｍatory cytokines are evaluated by ELISA. Results: Immunohistochemistry showed increased expression of TLR4 and 5 in placental tissue of CAM (+) samples. But TLR 2 or 6 didn’t show significant difference between CAM(+) and CAM(-) groups. IL6/8 also strongly expressed in the same site in which TLR5 was stained. In the tissue culture, IL6 expression was remarkably increased by TLR2/4/5 ligands. But no significant difference of IL8 was seen. Conclusions: This study shows that, in chorioamnionitis, TLRs expression are elevated in the fetal membrane, and it may provide increased proinflammatory cytokine production. Therefore TLRs may pray important roles in human chorioamnionitis.
O-104. DNA METHYLATION ANALYSIS OF A CPG-RICH PROMOTER REGION OF C19MC IN HUMAN TROPHOBLAST CELLS Hironori Takahashi a, b, Akihide Ohkuchi a, Tomoyuki Kuwata a, Rie Usui a, Kunio Kikuchi b, Shigeki Matsubara a, Mitsuaki Suzuki a, Toshiyuki Takeshita c, Toshihiro Takizawa b. a Department of Obstetrics and Gynecology, Jichi Medical University, Tochigi, Japan; b Department of Molecular Medicine and Anatomy, Nippon Medical School, Tokyo, Japan; c Department of Reproductive Medicine, Perinatology and Gynecologic Oncology, Nippon Medical School, Tokyo, Japan Objective: In the human, miRNAs derived from the miRNA cluster on human chromosome 19 (C19MC), a primate-specific miRNA cluster, have been demonstrated to exhibit a placenta-specific expression pattern. The C19MC is an imprinting gene and is exclusively expressed from a paternal