- Email: [email protected]
The expression of NF-AT (Nuclear factor of activated T cells) messenger RNA in maternal peripheral blood cells
TUESDAY.
SEPTEMBER
5
151
P2.13.06 CHANGES IN GENE EXPRESSION DURING PROGRESSION OF OVARIAN CARCINOMA .I Tamer, E. K&men, W. El Rifai, M. Seppiilii, Helsinki University Central Hospital, Haarhnaninkatu 2, Helsinki, Finland, 00290. Objectives: The molecular events leading to development and progression of serous ovarian carcinoma are incompletely understood. We carried out a large scale analysis of differentially expressed genes that might be associated with the development and progression of serous ovarian carcinoma. Study methods: The study population comprised five advanced and/or moderately or poorly differentiated serous adenocarcinomas, one local, highly differentiated serous adenocarcinoma, and one benign serous adenoma. cDNA array tehcnique was used to find differences in gene expression between serous adenocarcinoma and benign serous adenoma, and between advanced and/or moderately or poorly differentiated and local, highly differentiated serous adenocarcinoma. Results: The most striking difference between adenocarcinoma and benign adenoma were upregulation of ARHGDIB, MET and DLG3, and downregulation of HGFAC, DES, and PDGFR.4. The most prominent differences between advanced and local adenocarcinoma were upregulation of COL3A1, FGFRI and MET in advanced carcinoma, and, downregulation of HGFAC, FZD3, and BFLl in the same tumors. Conclusions: Significant differences in gene expression were found between malignant and benign serous ovarian tumors, and between advanced and/or moderately or poorly differentiated and local, highly differentiated serous adenocarcinomas. It is possible that the differentially expressed genes consistently found in malignant tumors may be associated with carcinogenesis and genes which were differentially expressed in advanced but not local carcinoma may play a role in tumor progression.
P2.14 PHYSIOLOGY
OF REPRODUCTION
2
P2.14.01 DEVELOPMENT CHANGES IN OXIDATIVE METABOLISM IN IMMATURE RAT BRAIN MITOCHONDRIA Y. Taniuchi, A. Nakai, H. Asakura, A. Yokota, T. Koshino, T. Araki, Dept. OB/GYN, Nippon Medical School, Tokyo, Japan. Objective: The present experiments were undertaken to investigate mitochondrial activity and energy metabolism in the developing rat brain from the late fetal stage to the neonatal stage. Methods: Samples of cerebral tissue were obtained from fetuses at 14, 16,lE and 20 days of gestation, and from pups at one hour, one day and 7 days after birth. Mitochondrial respiration was measured polarographically using hemogenates. Fetal and neonatal brains were frozen in situ and fluometric enzymatic techniques were used for the analysis of ATP, ADP and AMP. Results: In the fetal brain, there was a gradual increase in stimulated, and uncoupled respiratory rates from 14 days to 20 days of gestation, together with a moderate increase in ATP concentration and sum total of adenine nucleotides. Non-stimulated respiratory rates did not change with increasing gestational age. An increase in mitochondrial activity was more pronounced immediately after birth, together with a marked increase in high-energy phosphates. Conclusion: These results indicate that in the rat brain, there is maturation of oxidative metabolism in mitochondria that is initiated in late gestation. Acceleration in mitochondrial respiration occurs immediately after birth in order to maintain high-energy phosphate levels, and this may be crucial for the successful outcome of the newborn.
P2.14.02 THE EXPRESSION OF NF-AT (NUCLEAR FACTOR OF ACTIVATED T CELLS) MESSENGER RNA IN MATERNAL PERIPHERAL BLOOD CELLS H. Koiima, T. Tamura, T. Okuda, C. Kate, Y. Kinoshita, T. Yamamoto, Dept. OB/GYN, Kyoto Prefectural University of Medicine, Kyoto, Japan. Objectives: In T lymphocyte, NF-AT regulates the induction of immunoregulatory protein genes, including IL-2, THF-a and IL-4, with
the stimulation of antigens. This study was designed to analyze the association of NF-AT with pregnancy to see the systemic immunological changes in peripheral blood cells caused by pregnancy. Study Methods: With informed consent, peripheral blood was obtained from non-pregnant women with menstrual cycle (n=29) and pregnant women (n=239) and total RNA was prepared for RT. The relative expression ratios of NF-AT2 and NF-AT3 mRNAs against p-actin mRNA were measured by quantitative RT-PCR method using the specific primer pairs for the each objective gene. Results: In early pregnancy, the expression of NF-AT2 and NF-AT3 mRNAs were suppressed and in the first and second trimester of pregnancy they kept significantly lower levels than that in non-pregnant women (p
P2.14.03 THE EXPRESSION OF IL-8 AND IL-4 MESSENGER RNAS IN MATERNAL PERIPHERAL BLOOD CELLS DURING PREGNANCY C. T. Tamura, T. Okuda, H. Kojima, M. Fukuoka, T. Yamamoto, H. Honjo, Dept. OB/GYN, Kyoto Prefectural University of Medicine, Kyoto, Japan. Objectives: To see systemic immunological changes caused by pregnancy, we examined the expression of IL-18, IL-8 and IL-4 mRNAs in peripheral blood cells from pregnant and non-pregnant women by quantitative RTPCR method. Study Methods: With informed consent, peripheral blood was obtained from non-pregnant women with regular menstrual cycle (n=83) and pregnant women (n=56.5). Total RNA was prepared for R and the following quantitative PCR analysis. The relative expression ratio of IL18, IL-8 and IL=4 mRNAs against p-actin mRNA were measured. Results: In early pregnancy, the expression of IL-18 and IL-8 mRNAs were suppressed. They kept significantly lower levels than that in non-pregnant women from the 2”d till the Eth month of gestation (p
P2.14.04 THE EFFECTS OF OXYTOCIN ON THE PATTERN OF ELECTRICAL PROPAGATION IN THE ISOLATED PREGNANT UTERUS OF THE RAT. W.J.E.P. Lammers’, Betty Stephen’, R.HamidZ. D.W.G. Haroon3, ‘Dept. of Physiology, 3Dept. of Pharmacology, Faculty of Medicine & Health Sciences, UAE University, P.O. Box 17666, Al Ain, United Arab Emirates. ‘Department of Obstetrics & Gynecology, Faculty of Health Sciences, The Aga Khan University, Stadium Road, P.O. Box 3500, Karachi 74800 - Pakistan Objectives: To study the effects of Oxytocin at low and high concentrations, on the pattern of electrical propagation in the isolated pregnant uterus of the Rat. Study Methods: In the isolated pregnant myometrium of the rat, the pattern of propagation was investigated by recording simultaneously from 240 different extracellular sites while the contraction of the tissue was recorded isometrically. Analysis of all recorded electrograms allowed the twodimensional spread of activity in the myometrium to be reconstructed. From these activation maps, the conduction velocities were measured in the longitudinal, oblique and transversal directions. Results: At low concentrations (1Ct9 and 5~10~~IvQ,oxytocin significantly increased the frequency and duration of electrical bursts and the average
SEPTEMBER
5
151
P2.13.06 CHANGES IN GENE EXPRESSION DURING PROGRESSION OF OVARIAN CARCINOMA .I Tamer, E. K&men, W. El Rifai, M. Seppiilii, Helsinki University Central Hospital, Haarhnaninkatu 2, Helsinki, Finland, 00290. Objectives: The molecular events leading to development and progression of serous ovarian carcinoma are incompletely understood. We carried out a large scale analysis of differentially expressed genes that might be associated with the development and progression of serous ovarian carcinoma. Study methods: The study population comprised five advanced and/or moderately or poorly differentiated serous adenocarcinomas, one local, highly differentiated serous adenocarcinoma, and one benign serous adenoma. cDNA array tehcnique was used to find differences in gene expression between serous adenocarcinoma and benign serous adenoma, and between advanced and/or moderately or poorly differentiated and local, highly differentiated serous adenocarcinoma. Results: The most striking difference between adenocarcinoma and benign adenoma were upregulation of ARHGDIB, MET and DLG3, and downregulation of HGFAC, DES, and PDGFR.4. The most prominent differences between advanced and local adenocarcinoma were upregulation of COL3A1, FGFRI and MET in advanced carcinoma, and, downregulation of HGFAC, FZD3, and BFLl in the same tumors. Conclusions: Significant differences in gene expression were found between malignant and benign serous ovarian tumors, and between advanced and/or moderately or poorly differentiated and local, highly differentiated serous adenocarcinomas. It is possible that the differentially expressed genes consistently found in malignant tumors may be associated with carcinogenesis and genes which were differentially expressed in advanced but not local carcinoma may play a role in tumor progression.
P2.14 PHYSIOLOGY
OF REPRODUCTION
2
P2.14.01 DEVELOPMENT CHANGES IN OXIDATIVE METABOLISM IN IMMATURE RAT BRAIN MITOCHONDRIA Y. Taniuchi, A. Nakai, H. Asakura, A. Yokota, T. Koshino, T. Araki, Dept. OB/GYN, Nippon Medical School, Tokyo, Japan. Objective: The present experiments were undertaken to investigate mitochondrial activity and energy metabolism in the developing rat brain from the late fetal stage to the neonatal stage. Methods: Samples of cerebral tissue were obtained from fetuses at 14, 16,lE and 20 days of gestation, and from pups at one hour, one day and 7 days after birth. Mitochondrial respiration was measured polarographically using hemogenates. Fetal and neonatal brains were frozen in situ and fluometric enzymatic techniques were used for the analysis of ATP, ADP and AMP. Results: In the fetal brain, there was a gradual increase in stimulated, and uncoupled respiratory rates from 14 days to 20 days of gestation, together with a moderate increase in ATP concentration and sum total of adenine nucleotides. Non-stimulated respiratory rates did not change with increasing gestational age. An increase in mitochondrial activity was more pronounced immediately after birth, together with a marked increase in high-energy phosphates. Conclusion: These results indicate that in the rat brain, there is maturation of oxidative metabolism in mitochondria that is initiated in late gestation. Acceleration in mitochondrial respiration occurs immediately after birth in order to maintain high-energy phosphate levels, and this may be crucial for the successful outcome of the newborn.
P2.14.02 THE EXPRESSION OF NF-AT (NUCLEAR FACTOR OF ACTIVATED T CELLS) MESSENGER RNA IN MATERNAL PERIPHERAL BLOOD CELLS H. Koiima, T. Tamura, T. Okuda, C. Kate, Y. Kinoshita, T. Yamamoto, Dept. OB/GYN, Kyoto Prefectural University of Medicine, Kyoto, Japan. Objectives: In T lymphocyte, NF-AT regulates the induction of immunoregulatory protein genes, including IL-2, THF-a and IL-4, with
the stimulation of antigens. This study was designed to analyze the association of NF-AT with pregnancy to see the systemic immunological changes in peripheral blood cells caused by pregnancy. Study Methods: With informed consent, peripheral blood was obtained from non-pregnant women with menstrual cycle (n=29) and pregnant women (n=239) and total RNA was prepared for RT. The relative expression ratios of NF-AT2 and NF-AT3 mRNAs against p-actin mRNA were measured by quantitative RT-PCR method using the specific primer pairs for the each objective gene. Results: In early pregnancy, the expression of NF-AT2 and NF-AT3 mRNAs were suppressed and in the first and second trimester of pregnancy they kept significantly lower levels than that in non-pregnant women (p
P2.14.03 THE EXPRESSION OF IL-8 AND IL-4 MESSENGER RNAS IN MATERNAL PERIPHERAL BLOOD CELLS DURING PREGNANCY C. T. Tamura, T. Okuda, H. Kojima, M. Fukuoka, T. Yamamoto, H. Honjo, Dept. OB/GYN, Kyoto Prefectural University of Medicine, Kyoto, Japan. Objectives: To see systemic immunological changes caused by pregnancy, we examined the expression of IL-18, IL-8 and IL-4 mRNAs in peripheral blood cells from pregnant and non-pregnant women by quantitative RTPCR method. Study Methods: With informed consent, peripheral blood was obtained from non-pregnant women with regular menstrual cycle (n=83) and pregnant women (n=56.5). Total RNA was prepared for R and the following quantitative PCR analysis. The relative expression ratio of IL18, IL-8 and IL=4 mRNAs against p-actin mRNA were measured. Results: In early pregnancy, the expression of IL-18 and IL-8 mRNAs were suppressed. They kept significantly lower levels than that in non-pregnant women from the 2”d till the Eth month of gestation (p
P2.14.04 THE EFFECTS OF OXYTOCIN ON THE PATTERN OF ELECTRICAL PROPAGATION IN THE ISOLATED PREGNANT UTERUS OF THE RAT. W.J.E.P. Lammers’, Betty Stephen’, R.HamidZ. D.W.G. Haroon3, ‘Dept. of Physiology, 3Dept. of Pharmacology, Faculty of Medicine & Health Sciences, UAE University, P.O. Box 17666, Al Ain, United Arab Emirates. ‘Department of Obstetrics & Gynecology, Faculty of Health Sciences, The Aga Khan University, Stadium Road, P.O. Box 3500, Karachi 74800 - Pakistan Objectives: To study the effects of Oxytocin at low and high concentrations, on the pattern of electrical propagation in the isolated pregnant uterus of the Rat. Study Methods: In the isolated pregnant myometrium of the rat, the pattern of propagation was investigated by recording simultaneously from 240 different extracellular sites while the contraction of the tissue was recorded isometrically. Analysis of all recorded electrograms allowed the twodimensional spread of activity in the myometrium to be reconstructed. From these activation maps, the conduction velocities were measured in the longitudinal, oblique and transversal directions. Results: At low concentrations (1Ct9 and 5~10~~IvQ,oxytocin significantly increased the frequency and duration of electrical bursts and the average