- Email: [email protected]
Relationship between gestational age and frequency of fetal trophoblasts and nucleated erythrocytes in maternal peripheral blood
THURSDAY,
SEPTEMBER
7
(npIGFBP-1) have been identified in various biological fluids. To elucidate the biological effects of these phosphoisoforms, we studied effects of pIGFBP-1 and npIGFBP-1 on amino acid uptake induced by IG-I using cultured trophoblast cells. Study Methods: IGFBP-1 was purified from mid term amniotic fluid using ammonium sulfate precipitation followed by phenyl-Sepharose column. Purified IGFBP-1 was further purified by DEAE-cellulose column in which pIGFBP-1 and npIGFBP-1 were separated. Trophoblast cells obtained from term pregnancy were incubated with indicated concentration of pIGFBP-1 or npIGFBP-1 for 24 hr and further incubated with 10 nM IGF-I for 3 hr. Cells were then incubated with 3H-a-amino isobutyric acid (SH-AIB) for 30 min. Cells were solubilized and the radioactivity in cells was counted by a scintillation counter. Results: Both pIGFBP-1 and npIGFBP-1 alone had no effect on SH-AIB uptake, however, pIGFBP-1 inhibited IGF-I stimulated SH-AIB uptake with ED 50 of 0.26 nM while npIGFBP-1 potentiated SH-AIB uptake with ED50 of 0.27 nM. Maternal IGF-I promotes fetal growth by stimulating nutrients transport in the placenta. Conclusions: As shown in this study, pIGFBP-1 inhibits while npIGFBP-1 stimulates this IGF-I action in the placenta. Thus, it is suggested that IGFBP-1 phosphoisoforms are also involved in fetal growth by modulating IGF-I action in the placenta.
P4.16.14 PRENATAL DETERMINATION OF FETAL SEX AND RH D STATUS BY DNA ANALYSIS OF MATERNAL PLASMA K. Hiraki, H. Masuzaki, D. Nakayama, T. Ishimaru, Dept. OBIGYN, Nagasaki Univ Sch Med, Nagasaki, Japan Objectives: The recent demonstration of fetal DNA in maternal plasma raises the possibility that fetal DNA analysis may be possible with the use of maternal plasma, without cell separating techniques. We report use of PCR to detect fetal Y chromosome and Rh D gene from maternal plasma. Study Methods: We used QIAamp DNA Blood Mini Kit to extract DNA from maternal plasma. We used conventional PCR to amplify a Yspecific DNA sequence in plasma DNA samples from 26 pregnant women who had a gestational age of 14 to 41 weeks, and a Rh D sequence from three Rh D-negative pregnant women of 14 to 34 weeks. Results: Y sequences were detected in all of the 11 maternal plasma samples from women bearing male fetuses. None of the 15 women bearing female fetuses had positive results. The plasma samples obtained from three Rh D-negative women were positive for Rh D PCR analysis, which were concordant with the results of serologic analysis of the neonates. Conclusions: Prenatal determination of fetal sex and Rh D status with the use of maternal plasma can be performed rapidly and reliably. Fetal DNA in maternal plasma may be a valuable source of material for nonivasive prenatal diagnosis.
P4.16.15 PRELIMINARY RESULTS ON THE REGULATION OF THE HUMAN FSH RECEmOR PROMOTER ACTIVITY BY SELECTED E2F FAMILY MEMBERS L. Jakowicki (l), L. Putowski (l), C. Lee (2), W. Schillings (3), P. Reddy (2) (1) Dept. Surgical Gynecology, University School of Medicine, Lublin, Poland. (2) Fels Institute for Cancer Research and Molecular Biology, Philadelphia, PA, USA. (3) Women’s Health Care Associates Newton, NJ, USA. Objective: Since E2F’s transcriptional factors regulate expression of several proteins involved in cell cycle regulation we underwent studies concerning the influence of this factor on he FSH receptor promoter activity. Study Methods: DNA fragment containing FSH receptor promoter was sublcloned into pGL3 vector. Construct alone or together with expression vectors for E2F factor was transfected into cultured CHO cells. Additionally pRL-CMV vector was co-transfected in order to normalize transfection efficiency. Promoter activity was estimated by the measurement of firefly luciferase activity in cell lyzate. Obtained
143
results were normalized to the renilla activity driven by co-transfected pRL-CMV vector. Results: E2Fl decreased promoter activity (0,43*0,001). E2F4 and E2F5 increased promoter activity (1.29*0.04 and 1.66*0.20). Conclusions: Surprisingly increase (1.66 fold) in the activity caused by E2F5 is not so dramatic as it was noted previously for rat FSH-R promoter under the same condition. Lack of the E2F binding site in the human FSH-R receptor promoter, whereas it is present in rat gene, is possible explanation of these differences. E2Fl decreases the FSH-R promoter activity. It is more likely, the influence of E2F family members on human FSH-R promoter seems to be an indirect effect of this proteins.
P4.16.16 RELATION BETWEEN BIRTH WEIGm AND GENETIC POLYMORPHISM OF THE INSULIN GENE REGION IN JAPANESE Y. H. Osada, H. Sekimoto, K. Masuda, K. Seki, S. Sekiya, Dept. OBIGYN, Chiba University School of Medicine, Chiba, Japan. Objective: Fetal development depends not only on the intrauterine environment provided by the mother, but is closely associated with genetic factors from both parents. We examined the relation between birth weight and gene polymorphism in the IDDM2 region including the insulin gene in Japanese. Study Methods: We enrolled 211 neonates delivered to Japanese mothers after 36 weeks of normal pregnancy. DNA was extracted from the umbilical blood and also maternal peripheral blood cells. Polymorphic regions in the insulin-like growth factor II (IGF2) and tyrosine hydroxylase (TH) genes adjacent to the insulin gene were amplified by PCR. Birth weights were converted into standard deviation unit (SDU) based on gestational week, sex and para status. Results: For IGF2 polymorphism, SDU of birth weight were not significantly different among the 3 genotypes (-/-,+/-,+/+). For TH polymorphism the frequencies of neonates with alleles 6 to 11 were 25.9,24.9. 4.7,39.8,4.5 and 0.3%, respectively. Allele 10 has been reported to have strong linkage with class III allele that is a repeating sequence upstream of the insulin gene. The SDU was significantly higher in individuals with allele 10 than other alleles (p=O.O16). SDU were significantly different between neonates with and those without allele 10 in the genotype (p=O.O007). When classified further by parental origin of allele 10, a significant difference in SDU was observed only between paternal derived allele 10 (+) and allele 10 (-) groups (p=O.O027). Conclusion: This study showed that gene expression of the insulin gene region is one of the factors that regulate development in fetal stage.
P4.16.17 RELATIONSHIP BETWEEN GESTATIONAL AGE AND FREQUENCY OF FETAL TROPHOBLASTS AND NUCLEATED ERYTHROCYTES IN MATERNAL PERIPHERAL BLOOD A. Tan (l), T-H. Lim (l), V.H.H. Goh (2), (1) Singapore General Hospital, Outram Road, Singapore, 169608, (2) National University Hospital, Singapore. The relationship between gestational age and frequency of fetal cells in the maternal blood was studied in order to determine the optimal time for cell recovery. Immunomagnetic colloid system was used to enrich nucleated erythrocytes (NRBCs) and trophoblasts from 20ml of maternal blood obtained between 9 and 35 weeks gestation (N=41). Nested polymerase chain reaction (PCR) for the Y chromosome of enriched NRBCs and trophoblasts showed decreasing negative predictive values with increasing gestational age. The sensitivity and th overall frequency for correct fetal gender diagnosis were the lowest in the third trimester. Fluorescence in situ hybridisation (FISH) using XY DNA-specific probes was used to determine the fetal gender of trophoblasts-enriched fraction. The fetal origin of enriched NRBCs was determined using simultaneous immunophenotyping for fetal hemoglobin and FISH with XY probes. The mean number and mean percentage purity for both fetal trophoblasts and NRBCs showed decreasing values with increasing gestational age. However, statistical analysis showed no relationship between gestational age and frequency of fetal cells even though more fetal cells tend to exist during the first trimester. Nevertheless, the first
144
THURSDAY,
trimester appears to offer the most optimal time for fetal cell recovery from maternal blood for the purpose of prenatal diagnosis.
P4.16.18 SOMATIC MUTATIONS AND GENETIC POLYMORPHISMS OF THE PPPRlR3 GENE IN PATIENTS WITH SEVERAL TYPES OF CANCERS S. Takakura (11, A. Okamoto (2), .I. Yokota (l), T. Tanaka (2), T. Kohno
(l), T. Yamada (l), K. Shimizu (l), S. Ohwada (l), (1)National Cancer Center Research Institute, l-l, Tsukiji 5-chome, Chuo-ku, Tokyo, Japan, 1040045, (2) Jikei University School of Medicine, Tokyo, Japan. Objective: Recent studies on mEN and PPP2RlB (a regulatory subunit of PPZA) mutations in human cancer indicate that aberrations of intracellular signaling pathways via protein phosphatases (PP) are involved in human carcinogenesis. We examined genetic alterations of the PPPlR3 gene located at chromosome 7q31, which encodes the regulatory subunit 3 of PPl, in various types of human cancers to asses the role of this gene in human carcinogenesis. Study Methods: All of coding exons of PPPlR3 were examined for mutations and polymorphisms in 104 cancer cell lines and 192 primary tumors by PCR-SSCP and direct sequencing. Results: Sixteen mutations of the PPPlR3 gene were detected in 9 of 104 cancer cell lines and 5 of 192 primary tumors, and they occurred in a subset of ovarian cancer, lung cancer, colorectal cancer, and gastric cancer. Sixteen mutations detected consisted of 3 nonsense mutations, 10 missense mutations, 2 silent mutations and a mutation in intron sequence. Seven novel single nucleotide polymorphisms (SNPs) associated with the substitution of amino acids were also identified in cancer patients, in addition to three known nonsynonymous SNPs, including three previously reported ones as having an impact on the susceptibility to insulin resistant disorders. Conclusions: Differences in the activities and properties of multiple PPPlR3 proteins, which are produced in human cells due to variable somatic mutations and genetic polymorphisms in the PPPlR3 gene, can be involved in human carcinogenes
P4.16.19 f3,-GLYCOPROTEIN AND IMMUNE PLACENTAL TISSUES OF WOMEN PREGNANCY LOSS
COMPLEXES LOCATION WITH REPEATED
G.L.Gromiko. L.B.Zubjitskya, I.P.Pavlov Medical Univercity, D.O.Ott Institute of Obstetrics and Gynecology, St.Petersburg,
IN
Russia
Objectives: To study the consequence of antiphospholipid antibodies (aPL) on pregnancy complications we investigate localization of flzglycoprotein I @,-GP I) in placental tissues of women with recurrent pregnancy loss. Having a regulatory role in blood coagulation, fl,-GP I appear significantly related to most frequent complications (thrombosis and fetal loss) in patients with aPL. Study Methods: 46 samples of placentae and blood sera of women with repeated pregnancy losses were investigated. Method of fluorescent antibodies in indirect modification with sera of women that were tested on the presence of anti-cardiolipin antibodies on bovine heart was used. For elimination of cross-reacted antibodies to bovine cardiolipin and save antibodies to fl,-GP I tested sera were adsorbed by standart cardiolipin antigen. Placentae cryostat sections of women were treated for detecting complement-fixing immune complexes (IC). In parallel after washing, placentae sections were treated by adsorbede sera containing fl,-GP I. Results: Specific luminescence of IC was detected in 91% of women, aPL in 78%. Luminescence was detected on membrans of syncytiotrophoblast and on endothelium of chorion vessels. The topographical identity of a luminescence of complement-fixing IC is established with sites of a luminescences, obtained at handling washing placentae sections by adsorbede sera. Conclusions: We assume presence of the fl,-GP I in a structure of IC localized in a placental tissue. Being an antigenic target of aPL, fl,-GP I can be immediately connected to development of placental thrombosis
SEPTEMBER
P4.16.20 THE APPLICATION GENNO-ENGINEERING azp- INTERFERON THE TREATMENT NONSPECIFIC INFLAMMATORY DISEASES OF WOMEN’S REPRODUCTION ORGANS T.V. Bannvcova (l), T.N. Dyomina (2), G.D. Mysuna (2),
(1) (2)
7
IN
Donetsk State Regional Centre Gruarding Maternity and Childhood, Donetsk, Panfilova, Ukraine. Donetsk State Medical University, Done&k, Illycha, Ukraine.
Objectives: The aim of given research was the study of the effect gennoengineering ~1~~ interferon during treatment nonspecific inflammatory diseases of women’s internal genitals. Study Methods: It was conducted clinic-immunology investigation 108 women with nonspecific inflammatory diseases of ovaries and of uterine appendages. The 70 patients had immunocorrection therapy by “Laferon” (preparation ~1~~ interferon), the 40 patients -had the traditional antibacterial treatment. Results: The suppresion cell and humoral links of the immunity (the deficit T-cell, the dysimmunoglobulynemia), the factors of the nonspecific stability (the reduction of metabolic potence leukocytes) the interferon-deficit state (the higher level serum interferon’s, the reduction of the production the direct CIand y interferon) was revealed in patients of two groups. It was marked the normalization of functions the immunocompetents system’s in the group, who was treated by the “Laferon.” The convalescence was coming in 97.1% of the patients, it was marked improvement of state considerable 2.9% of patients. The positive dynamics of the index immunity wasn’t marked in the patient, who had antibacterial therapy: 72.5% women has convalescence; the state has improved in 20%; the therapy was not effective for 7.5% of the patients. Conclusions: The using a preparation ~1~~ interferon to allow back to normalcy the functions of the immunocompetent systems’ and to raise the effectiveness therapy nonspecific inflammatory diseases of internal genitals females.
P4.16.21 THE CORRECTION OF THE HORMONAL INSUFFICIENCY OF THE OVARIES WITH THE TRANSPLANTATION OF THE OVARIAN TISSUE L. Dept. PED/AD GYN, Done&k regional center of Maternity
and Child Protection, Done&k, Ukraine. Objective: The task of our invention is the correction of the hormonal insufficiency of the ovaries by the transplantation of the culture of the ovarian tissue cells. Study Methods: A new thing of this method is that the culture of the cells of the ovarian tissue is used as a transplant. The safety and the functional ability of the transplant is a main condition of the successful hormonal correction of the sufficient function of the ovaries. Results: The cells of the fetus ovarian tissue, which do not have an antigen activity, have not undergone the rejection, function on being introduced into the organism, producing estradiol and progesterone, in connection with this it appeared a possibility with the help of allotransplantation to correct the insufficiency of the ovarian function. Effective functioning transplanted culture of the ovarian tissue and absorption of estrogens, produced by it, is possible due to the increased adequate and sufficient permeability through the walls newly formed capsule. Conclusions: Thus, the quality of the introduced material was improved, the permeability throng the formed capsule was increased, the duration of the functioning of the transplant cells was increased, the culture in the form of suspension was introduced and activation of blood supply round the formed capsule, contributing to the formation and appearance of estrogens, lead to the start of the mechanisms of H-H-O regulation and initiation of extragonadal synthesis of estrogens and progesterone.
P4.16.22 THE ROLE OF AMNIOTIC FLUID INTERPHASE FISH ANALYSIS IN PATIENT MANAGEMENT W.C. Leung (1,2), E.J.T. Winsor (l), G. Seaward (l), R. Windrim (l),
D. Chitayaat (l), G. Ryan (1) (1) University of Toronto Perinatal Complex, Toronto, Ontario, Canada. (2) Dept. OBIGYN, University of Hong Kong, Hong Kong, China.
SEPTEMBER
7
(npIGFBP-1) have been identified in various biological fluids. To elucidate the biological effects of these phosphoisoforms, we studied effects of pIGFBP-1 and npIGFBP-1 on amino acid uptake induced by IG-I using cultured trophoblast cells. Study Methods: IGFBP-1 was purified from mid term amniotic fluid using ammonium sulfate precipitation followed by phenyl-Sepharose column. Purified IGFBP-1 was further purified by DEAE-cellulose column in which pIGFBP-1 and npIGFBP-1 were separated. Trophoblast cells obtained from term pregnancy were incubated with indicated concentration of pIGFBP-1 or npIGFBP-1 for 24 hr and further incubated with 10 nM IGF-I for 3 hr. Cells were then incubated with 3H-a-amino isobutyric acid (SH-AIB) for 30 min. Cells were solubilized and the radioactivity in cells was counted by a scintillation counter. Results: Both pIGFBP-1 and npIGFBP-1 alone had no effect on SH-AIB uptake, however, pIGFBP-1 inhibited IGF-I stimulated SH-AIB uptake with ED 50 of 0.26 nM while npIGFBP-1 potentiated SH-AIB uptake with ED50 of 0.27 nM. Maternal IGF-I promotes fetal growth by stimulating nutrients transport in the placenta. Conclusions: As shown in this study, pIGFBP-1 inhibits while npIGFBP-1 stimulates this IGF-I action in the placenta. Thus, it is suggested that IGFBP-1 phosphoisoforms are also involved in fetal growth by modulating IGF-I action in the placenta.
P4.16.14 PRENATAL DETERMINATION OF FETAL SEX AND RH D STATUS BY DNA ANALYSIS OF MATERNAL PLASMA K. Hiraki, H. Masuzaki, D. Nakayama, T. Ishimaru, Dept. OBIGYN, Nagasaki Univ Sch Med, Nagasaki, Japan Objectives: The recent demonstration of fetal DNA in maternal plasma raises the possibility that fetal DNA analysis may be possible with the use of maternal plasma, without cell separating techniques. We report use of PCR to detect fetal Y chromosome and Rh D gene from maternal plasma. Study Methods: We used QIAamp DNA Blood Mini Kit to extract DNA from maternal plasma. We used conventional PCR to amplify a Yspecific DNA sequence in plasma DNA samples from 26 pregnant women who had a gestational age of 14 to 41 weeks, and a Rh D sequence from three Rh D-negative pregnant women of 14 to 34 weeks. Results: Y sequences were detected in all of the 11 maternal plasma samples from women bearing male fetuses. None of the 15 women bearing female fetuses had positive results. The plasma samples obtained from three Rh D-negative women were positive for Rh D PCR analysis, which were concordant with the results of serologic analysis of the neonates. Conclusions: Prenatal determination of fetal sex and Rh D status with the use of maternal plasma can be performed rapidly and reliably. Fetal DNA in maternal plasma may be a valuable source of material for nonivasive prenatal diagnosis.
P4.16.15 PRELIMINARY RESULTS ON THE REGULATION OF THE HUMAN FSH RECEmOR PROMOTER ACTIVITY BY SELECTED E2F FAMILY MEMBERS L. Jakowicki (l), L. Putowski (l), C. Lee (2), W. Schillings (3), P. Reddy (2) (1) Dept. Surgical Gynecology, University School of Medicine, Lublin, Poland. (2) Fels Institute for Cancer Research and Molecular Biology, Philadelphia, PA, USA. (3) Women’s Health Care Associates Newton, NJ, USA. Objective: Since E2F’s transcriptional factors regulate expression of several proteins involved in cell cycle regulation we underwent studies concerning the influence of this factor on he FSH receptor promoter activity. Study Methods: DNA fragment containing FSH receptor promoter was sublcloned into pGL3 vector. Construct alone or together with expression vectors for E2F factor was transfected into cultured CHO cells. Additionally pRL-CMV vector was co-transfected in order to normalize transfection efficiency. Promoter activity was estimated by the measurement of firefly luciferase activity in cell lyzate. Obtained
143
results were normalized to the renilla activity driven by co-transfected pRL-CMV vector. Results: E2Fl decreased promoter activity (0,43*0,001). E2F4 and E2F5 increased promoter activity (1.29*0.04 and 1.66*0.20). Conclusions: Surprisingly increase (1.66 fold) in the activity caused by E2F5 is not so dramatic as it was noted previously for rat FSH-R promoter under the same condition. Lack of the E2F binding site in the human FSH-R receptor promoter, whereas it is present in rat gene, is possible explanation of these differences. E2Fl decreases the FSH-R promoter activity. It is more likely, the influence of E2F family members on human FSH-R promoter seems to be an indirect effect of this proteins.
P4.16.16 RELATION BETWEEN BIRTH WEIGm AND GENETIC POLYMORPHISM OF THE INSULIN GENE REGION IN JAPANESE Y. H. Osada, H. Sekimoto, K. Masuda, K. Seki, S. Sekiya, Dept. OBIGYN, Chiba University School of Medicine, Chiba, Japan. Objective: Fetal development depends not only on the intrauterine environment provided by the mother, but is closely associated with genetic factors from both parents. We examined the relation between birth weight and gene polymorphism in the IDDM2 region including the insulin gene in Japanese. Study Methods: We enrolled 211 neonates delivered to Japanese mothers after 36 weeks of normal pregnancy. DNA was extracted from the umbilical blood and also maternal peripheral blood cells. Polymorphic regions in the insulin-like growth factor II (IGF2) and tyrosine hydroxylase (TH) genes adjacent to the insulin gene were amplified by PCR. Birth weights were converted into standard deviation unit (SDU) based on gestational week, sex and para status. Results: For IGF2 polymorphism, SDU of birth weight were not significantly different among the 3 genotypes (-/-,+/-,+/+). For TH polymorphism the frequencies of neonates with alleles 6 to 11 were 25.9,24.9. 4.7,39.8,4.5 and 0.3%, respectively. Allele 10 has been reported to have strong linkage with class III allele that is a repeating sequence upstream of the insulin gene. The SDU was significantly higher in individuals with allele 10 than other alleles (p=O.O16). SDU were significantly different between neonates with and those without allele 10 in the genotype (p=O.O007). When classified further by parental origin of allele 10, a significant difference in SDU was observed only between paternal derived allele 10 (+) and allele 10 (-) groups (p=O.O027). Conclusion: This study showed that gene expression of the insulin gene region is one of the factors that regulate development in fetal stage.
P4.16.17 RELATIONSHIP BETWEEN GESTATIONAL AGE AND FREQUENCY OF FETAL TROPHOBLASTS AND NUCLEATED ERYTHROCYTES IN MATERNAL PERIPHERAL BLOOD A. Tan (l), T-H. Lim (l), V.H.H. Goh (2), (1) Singapore General Hospital, Outram Road, Singapore, 169608, (2) National University Hospital, Singapore. The relationship between gestational age and frequency of fetal cells in the maternal blood was studied in order to determine the optimal time for cell recovery. Immunomagnetic colloid system was used to enrich nucleated erythrocytes (NRBCs) and trophoblasts from 20ml of maternal blood obtained between 9 and 35 weeks gestation (N=41). Nested polymerase chain reaction (PCR) for the Y chromosome of enriched NRBCs and trophoblasts showed decreasing negative predictive values with increasing gestational age. The sensitivity and th overall frequency for correct fetal gender diagnosis were the lowest in the third trimester. Fluorescence in situ hybridisation (FISH) using XY DNA-specific probes was used to determine the fetal gender of trophoblasts-enriched fraction. The fetal origin of enriched NRBCs was determined using simultaneous immunophenotyping for fetal hemoglobin and FISH with XY probes. The mean number and mean percentage purity for both fetal trophoblasts and NRBCs showed decreasing values with increasing gestational age. However, statistical analysis showed no relationship between gestational age and frequency of fetal cells even though more fetal cells tend to exist during the first trimester. Nevertheless, the first
144
THURSDAY,
trimester appears to offer the most optimal time for fetal cell recovery from maternal blood for the purpose of prenatal diagnosis.
P4.16.18 SOMATIC MUTATIONS AND GENETIC POLYMORPHISMS OF THE PPPRlR3 GENE IN PATIENTS WITH SEVERAL TYPES OF CANCERS S. Takakura (11, A. Okamoto (2), .I. Yokota (l), T. Tanaka (2), T. Kohno
(l), T. Yamada (l), K. Shimizu (l), S. Ohwada (l), (1)National Cancer Center Research Institute, l-l, Tsukiji 5-chome, Chuo-ku, Tokyo, Japan, 1040045, (2) Jikei University School of Medicine, Tokyo, Japan. Objective: Recent studies on mEN and PPP2RlB (a regulatory subunit of PPZA) mutations in human cancer indicate that aberrations of intracellular signaling pathways via protein phosphatases (PP) are involved in human carcinogenesis. We examined genetic alterations of the PPPlR3 gene located at chromosome 7q31, which encodes the regulatory subunit 3 of PPl, in various types of human cancers to asses the role of this gene in human carcinogenesis. Study Methods: All of coding exons of PPPlR3 were examined for mutations and polymorphisms in 104 cancer cell lines and 192 primary tumors by PCR-SSCP and direct sequencing. Results: Sixteen mutations of the PPPlR3 gene were detected in 9 of 104 cancer cell lines and 5 of 192 primary tumors, and they occurred in a subset of ovarian cancer, lung cancer, colorectal cancer, and gastric cancer. Sixteen mutations detected consisted of 3 nonsense mutations, 10 missense mutations, 2 silent mutations and a mutation in intron sequence. Seven novel single nucleotide polymorphisms (SNPs) associated with the substitution of amino acids were also identified in cancer patients, in addition to three known nonsynonymous SNPs, including three previously reported ones as having an impact on the susceptibility to insulin resistant disorders. Conclusions: Differences in the activities and properties of multiple PPPlR3 proteins, which are produced in human cells due to variable somatic mutations and genetic polymorphisms in the PPPlR3 gene, can be involved in human carcinogenes
P4.16.19 f3,-GLYCOPROTEIN AND IMMUNE PLACENTAL TISSUES OF WOMEN PREGNANCY LOSS
COMPLEXES LOCATION WITH REPEATED
G.L.Gromiko. L.B.Zubjitskya, I.P.Pavlov Medical Univercity, D.O.Ott Institute of Obstetrics and Gynecology, St.Petersburg,
IN
Russia
Objectives: To study the consequence of antiphospholipid antibodies (aPL) on pregnancy complications we investigate localization of flzglycoprotein I @,-GP I) in placental tissues of women with recurrent pregnancy loss. Having a regulatory role in blood coagulation, fl,-GP I appear significantly related to most frequent complications (thrombosis and fetal loss) in patients with aPL. Study Methods: 46 samples of placentae and blood sera of women with repeated pregnancy losses were investigated. Method of fluorescent antibodies in indirect modification with sera of women that were tested on the presence of anti-cardiolipin antibodies on bovine heart was used. For elimination of cross-reacted antibodies to bovine cardiolipin and save antibodies to fl,-GP I tested sera were adsorbed by standart cardiolipin antigen. Placentae cryostat sections of women were treated for detecting complement-fixing immune complexes (IC). In parallel after washing, placentae sections were treated by adsorbede sera containing fl,-GP I. Results: Specific luminescence of IC was detected in 91% of women, aPL in 78%. Luminescence was detected on membrans of syncytiotrophoblast and on endothelium of chorion vessels. The topographical identity of a luminescence of complement-fixing IC is established with sites of a luminescences, obtained at handling washing placentae sections by adsorbede sera. Conclusions: We assume presence of the fl,-GP I in a structure of IC localized in a placental tissue. Being an antigenic target of aPL, fl,-GP I can be immediately connected to development of placental thrombosis
SEPTEMBER
P4.16.20 THE APPLICATION GENNO-ENGINEERING azp- INTERFERON THE TREATMENT NONSPECIFIC INFLAMMATORY DISEASES OF WOMEN’S REPRODUCTION ORGANS T.V. Bannvcova (l), T.N. Dyomina (2), G.D. Mysuna (2),
(1) (2)
7
IN
Donetsk State Regional Centre Gruarding Maternity and Childhood, Donetsk, Panfilova, Ukraine. Donetsk State Medical University, Done&k, Illycha, Ukraine.
Objectives: The aim of given research was the study of the effect gennoengineering ~1~~ interferon during treatment nonspecific inflammatory diseases of women’s internal genitals. Study Methods: It was conducted clinic-immunology investigation 108 women with nonspecific inflammatory diseases of ovaries and of uterine appendages. The 70 patients had immunocorrection therapy by “Laferon” (preparation ~1~~ interferon), the 40 patients -had the traditional antibacterial treatment. Results: The suppresion cell and humoral links of the immunity (the deficit T-cell, the dysimmunoglobulynemia), the factors of the nonspecific stability (the reduction of metabolic potence leukocytes) the interferon-deficit state (the higher level serum interferon’s, the reduction of the production the direct CIand y interferon) was revealed in patients of two groups. It was marked the normalization of functions the immunocompetents system’s in the group, who was treated by the “Laferon.” The convalescence was coming in 97.1% of the patients, it was marked improvement of state considerable 2.9% of patients. The positive dynamics of the index immunity wasn’t marked in the patient, who had antibacterial therapy: 72.5% women has convalescence; the state has improved in 20%; the therapy was not effective for 7.5% of the patients. Conclusions: The using a preparation ~1~~ interferon to allow back to normalcy the functions of the immunocompetent systems’ and to raise the effectiveness therapy nonspecific inflammatory diseases of internal genitals females.
P4.16.21 THE CORRECTION OF THE HORMONAL INSUFFICIENCY OF THE OVARIES WITH THE TRANSPLANTATION OF THE OVARIAN TISSUE L. Dept. PED/AD GYN, Done&k regional center of Maternity
and Child Protection, Done&k, Ukraine. Objective: The task of our invention is the correction of the hormonal insufficiency of the ovaries by the transplantation of the culture of the ovarian tissue cells. Study Methods: A new thing of this method is that the culture of the cells of the ovarian tissue is used as a transplant. The safety and the functional ability of the transplant is a main condition of the successful hormonal correction of the sufficient function of the ovaries. Results: The cells of the fetus ovarian tissue, which do not have an antigen activity, have not undergone the rejection, function on being introduced into the organism, producing estradiol and progesterone, in connection with this it appeared a possibility with the help of allotransplantation to correct the insufficiency of the ovarian function. Effective functioning transplanted culture of the ovarian tissue and absorption of estrogens, produced by it, is possible due to the increased adequate and sufficient permeability through the walls newly formed capsule. Conclusions: Thus, the quality of the introduced material was improved, the permeability throng the formed capsule was increased, the duration of the functioning of the transplant cells was increased, the culture in the form of suspension was introduced and activation of blood supply round the formed capsule, contributing to the formation and appearance of estrogens, lead to the start of the mechanisms of H-H-O regulation and initiation of extragonadal synthesis of estrogens and progesterone.
P4.16.22 THE ROLE OF AMNIOTIC FLUID INTERPHASE FISH ANALYSIS IN PATIENT MANAGEMENT W.C. Leung (1,2), E.J.T. Winsor (l), G. Seaward (l), R. Windrim (l),
D. Chitayaat (l), G. Ryan (1) (1) University of Toronto Perinatal Complex, Toronto, Ontario, Canada. (2) Dept. OBIGYN, University of Hong Kong, Hong Kong, China.