- Email: [email protected]
The action of angiotensin II and brain natriuretic peptide on the normal and diabetic placental vasculature
310
SPO Abstracts
J a n u a r y 1995 Am J Obstet Gynecol
177
IMMUNOREACTIVE PARATHYROID HORMONE-RELATED PEPTIDE: ITS ASSOCIATION WITH PRETERM LABOUR. M.M. Ramlrez., L. Fraher', D. Goftzma#, G. N. Handy', J.R.G. Challis'. Lawson Research Institute, MRC, Univ. of W, Ontado and IvlcGill Univ. OBJECTIVE: Parathyroid horrnone-related paptlde (PTHrP) is a 141 amino add protein that can be processed to a 1-34 n-terminal fragment that resembles Parathyroid hormone and has smooth muscle relaxant activity, and a mid (67-86) fragment which reportedly alters placental calcium transport. We used specific antibodies to these regions of PTHrP to determine patterns of localization of the psptides, relative changes in distribution and intensity of immunoatsining dudng term and praterm labour. STUDY DESIGN: Placental and fetal membranes were collected from patients with preterm delivery (PTL) (n,16), term cesarean section (n,lO) and term vaginal delivery In,.5). Immunohistochemistry was performed with spedflc antlsera, visualized with avidin-blotin method and staining intensity was quantified with the MCID Image Intensity Analyzer. RESULTS: Immunoresctive (IR) PTHrP (1-34) and (67-86) was localized in the amnion epithelium, chorionlc trophoblast, decidual cells and placental syncifiotrophoblast. Intense tmmunoataining was observed with PTHrP (67°86) but not with (1o34) in the endothelial lining of the villous cspillades. IR-PTHrP (1-34) was decreased in PTL in placenta and fatal membranes compared to vaginal delivery (p < 0.05 Mann-Witney test). After 24 hours of rupture of membranes there was a trend with decreased Immunoatainlng in PTL as well as term delivedes. In contrast, there was no statistical difference in the IR-PTHrP (68-86) within delivery categories. CONCLUSIONS: IR-PTHrP (1-34) is decreased In PTL compared to term delivery. The data suggest a temporal relationship between decreasing IR and pratarm delivery. The different pattern of IR between PTHrP (1-34) and (67-86) within placental villi, suggests a diversity in the paptide processing and / or catabolism.
179
HQRPHOHETRICSTUDYOF HUI41ANPLACENTALVESSELS: RELATIONSHIPWITH UNBILICAL A~ERY DODPLER FLOW. S.C. Hitra a, V.S. Venkataseshan~'~, J. Git°~ adept. Ob/Gyn, T-'h-e-eJama-i-~ecaHospitat, Jamaica, NY, bDept. PathoLogy, Ht. Sinai School of Hedieine, New York~ NY. OBJECTIVE: (1) To determine the changes of vessel waLL thickness and radius of Lumenof the tertiary stem viLLi of the placenta with advancing gastational ages and (2) their relationship to DoppLer flow of the umbiLicaL artery. STUDY DESIGN: F i f t y three placentas from preterm and normal pregnancies and deliveries (between 20 and 40 weeks) were used for morphometric study of the tertiary stem viLLi vessels. Each patient had DoppLer flow study of the umbiLicaL artery using pulse wave DoppLer and the resistance index (R]) was determined from DoppLer flow velocity waveform. RI values were compared with normal RI values (previousLy determined) for the corresponding gestational age. 4 micron paraffin sections of the placentas were stained with hematoxytin and eosin and periodic acid schiff reagent. The tertiary stem viLLi and i t s vessels were examined under Light Ix40) microscope, interactive morphemetry was performed using Intet (System 10) 286 microprocessor with math co-processor and a touch sensitive screen. VascuLar watt thickness was determined from the outer and inner circumferences (Lumen) of the vessel waLL. RESULTS: Thickness, NOT the lumen, of the t e r t i a r y stem viLLi vessels walt, had a significant negative Linear association (Y=33.719709-O.5227726*X, p
178
MECONIUM INHIBITS THE CONTRACTIONOF UMBILICALVESSELS INDUCEDBY THE THROMBOXANEAmANALOGUEU 46619. L Mnntonmorv.' M Belfort,'G Saade,'K Moise,Jr., Y Vndemikov.'Dept.OBIGYN,BeylerCollegeof Medicine,Houston,TX. OBJECTIVE: Maceniumhas beenshownto penetrateWherten'sjelly. In addition,thereis evidencethat macnoinmmaybe a vasoconstrictor. Ourobjectivewas to studythe direct effectsof maconium on isolatedumbilicalartery(UA)andvein(U~V)in.vitro. STUDY DESIGN: IntactUA andUVringsweresuspendedin 5 ml organbathscontaining Kreb'ssolution(pH 7.2; 37°C;bubbledwith 2.5% O~+ 8.07% CO2/ balanceN2)for isometrictensionrecording.Meneninm(finalbathconcentration- 1%),centrifugedand filteredmecenium,and mecnoiumwith the lipid fractionremoved(separatedinto < and > 30,000 KD),was addedto the baths. Somevesselswereincubatedin 1% maceniwnfor 30 minonteswhichwas thenwashedout. Concentration-responsocurvesto U46619 were obtained.TheEChoHog of the concentrationwhichevokes50% of the maximal contraction)was dcterndnnd.Analysis:ANOVA(with nunn's test), Significance;p < 0.05. S,o~ ~ . .u. .m. ~1%I RESULTS: IEC_.) UAInl UVIn, ~t29, ~w'ld Control 8.1+0.3(16) 8.0+0.4(11) ~ ~oo] / ~ ; : ~ Maconium1% 7.4+ 0.2" (10) 7.3+0.3"(11) :E =~ Meconium(washed) 7.9+0.4m(8) 7.6+0.4(9) ~ 6o Meconium(filtered) 7.7+0.3* (9) 7.4+0.5*(6) ~,o -6to Lipid Removed ~ o . . . . . . -s5 4o -r.5 -7o < 30.000 KD 7.9+0.4m(8) NotDono .8o U4eSlQnoaM1 > 3D.000 KD 8.0+ 0.2m(61 Not Done * - differentfromcontrol(p< 0.05); m, differentfrommaconium1% (p< 0.05) UA andUVhadno sustainedresponseto mecnoium.Tensiondevelopedby ringsexposedto meconiumwas significantlylessthancontrolat all concentrationsof U46619. Therewas somelossof the efficacyOfmeconiumfollowingcentrifugation/fifirationand washingout. Meconiumwithoutthe lipid fractionhadlessinhibitoryeffect than nativemeconium CONCLUSION: Meconiumdoesnot causesustainedcontractionof UA or UVin-vitro. It does,however,inhibitthe contractileeffect of U46819in both UA and UVsmoothmuscle. TheinhibitoryactivityQfmoconiumappearsto be concentratedin the lipidfraction.
180
LOCALIZATION OF NITRIC OXIDE SYNTHASE ACTIVITY IN VARIOUS COMPARTMENTS OF HUMAN PLACENTA. P.F. Mentox, P.I. Karlx, E.P. Girardix, B.M. Pctrikovsky and B.M. Wilkesx. Depts of Nephrology, Pediatrics and Fetal-Maternal Medicine, North Shore Univ Hospital-Comell Univ Med College, Manhassct, NY OBJECTIVE: We tested the hypothesis that nitric oxide synthas¢ (NOS) activity is differentially distributed in the placenta and preferentially expressed in the vascular compartment. STUDY DESIGN: Tissues from vascular and non-vascular compartments of 12 normal placentas were homogenized and separated into cytosolic and membrane fractions by ultraccntrifugation. NOS activity was measured by conversion of [14C]-arginine to [14C]-citrulline. Activity in wholo placental homogcnat¢ was compared with activity in human umbilical vein endothelial cells (HUVEC), small placental arteries (<1.0 mm diameter) and syncytiotrophoblasts in culture. RESULTS: In whole placenta, HUVECs, and small placental arteries, 80% of the NOS activity was in the membrane fraction with the remaining activity present in the cytosol. In contrast, in cultured syncytiotrophoblasts NOS activity occurred exclusively in the membrane fraction. NADPHdiaphorase staining of whole placenta was found only in the syncytiotrophoblasts. Day O isolated trophoblasts showed little diaphorasc staining and NOS activity. On days I and 3, as syncytiotrophoblasts differentiated, diaphoras¢ staining and NOS activity increased in parallel. CONCLUSIONS: The cellular distribution of nitric oxide synthas¢ activity was different in vascular and non-vascular compartments s!nce acitivity in syncytiotrophoblasts occurred exclusively in the membrane fraction. Nitric oxide synthesis increases as trophoblasts mature. These data suggest a potential role for nitric oxide in platelet adhesion in the intervillous space and regulation of resistance in fetal arterioles.
Y
SPO Abstracts
J a n u a r y 1995 Am J Obstet Gynecol
177
IMMUNOREACTIVE PARATHYROID HORMONE-RELATED PEPTIDE: ITS ASSOCIATION WITH PRETERM LABOUR. M.M. Ramlrez., L. Fraher', D. Goftzma#, G. N. Handy', J.R.G. Challis'. Lawson Research Institute, MRC, Univ. of W, Ontado and IvlcGill Univ. OBJECTIVE: Parathyroid horrnone-related paptlde (PTHrP) is a 141 amino add protein that can be processed to a 1-34 n-terminal fragment that resembles Parathyroid hormone and has smooth muscle relaxant activity, and a mid (67-86) fragment which reportedly alters placental calcium transport. We used specific antibodies to these regions of PTHrP to determine patterns of localization of the psptides, relative changes in distribution and intensity of immunoatsining dudng term and praterm labour. STUDY DESIGN: Placental and fetal membranes were collected from patients with preterm delivery (PTL) (n,16), term cesarean section (n,lO) and term vaginal delivery In,.5). Immunohistochemistry was performed with spedflc antlsera, visualized with avidin-blotin method and staining intensity was quantified with the MCID Image Intensity Analyzer. RESULTS: Immunoresctive (IR) PTHrP (1-34) and (67-86) was localized in the amnion epithelium, chorionlc trophoblast, decidual cells and placental syncifiotrophoblast. Intense tmmunoataining was observed with PTHrP (67°86) but not with (1o34) in the endothelial lining of the villous cspillades. IR-PTHrP (1-34) was decreased in PTL in placenta and fatal membranes compared to vaginal delivery (p < 0.05 Mann-Witney test). After 24 hours of rupture of membranes there was a trend with decreased Immunoatainlng in PTL as well as term delivedes. In contrast, there was no statistical difference in the IR-PTHrP (68-86) within delivery categories. CONCLUSIONS: IR-PTHrP (1-34) is decreased In PTL compared to term delivery. The data suggest a temporal relationship between decreasing IR and pratarm delivery. The different pattern of IR between PTHrP (1-34) and (67-86) within placental villi, suggests a diversity in the paptide processing and / or catabolism.
179
HQRPHOHETRICSTUDYOF HUI41ANPLACENTALVESSELS: RELATIONSHIPWITH UNBILICAL A~ERY DODPLER FLOW. S.C. Hitra a, V.S. Venkataseshan~'~, J. Git°~ adept. Ob/Gyn, T-'h-e-eJama-i-~ecaHospitat, Jamaica, NY, bDept. PathoLogy, Ht. Sinai School of Hedieine, New York~ NY. OBJECTIVE: (1) To determine the changes of vessel waLL thickness and radius of Lumenof the tertiary stem viLLi of the placenta with advancing gastational ages and (2) their relationship to DoppLer flow of the umbiLicaL artery. STUDY DESIGN: F i f t y three placentas from preterm and normal pregnancies and deliveries (between 20 and 40 weeks) were used for morphometric study of the tertiary stem viLLi vessels. Each patient had DoppLer flow study of the umbiLicaL artery using pulse wave DoppLer and the resistance index (R]) was determined from DoppLer flow velocity waveform. RI values were compared with normal RI values (previousLy determined) for the corresponding gestational age. 4 micron paraffin sections of the placentas were stained with hematoxytin and eosin and periodic acid schiff reagent. The tertiary stem viLLi and i t s vessels were examined under Light Ix40) microscope, interactive morphemetry was performed using Intet (System 10) 286 microprocessor with math co-processor and a touch sensitive screen. VascuLar watt thickness was determined from the outer and inner circumferences (Lumen) of the vessel waLL. RESULTS: Thickness, NOT the lumen, of the t e r t i a r y stem viLLi vessels walt, had a significant negative Linear association (Y=33.719709-O.5227726*X, p
178
MECONIUM INHIBITS THE CONTRACTIONOF UMBILICALVESSELS INDUCEDBY THE THROMBOXANEAmANALOGUEU 46619. L Mnntonmorv.' M Belfort,'G Saade,'K Moise,Jr., Y Vndemikov.'Dept.OBIGYN,BeylerCollegeof Medicine,Houston,TX. OBJECTIVE: Maceniumhas beenshownto penetrateWherten'sjelly. In addition,thereis evidencethat macnoinmmaybe a vasoconstrictor. Ourobjectivewas to studythe direct effectsof maconium on isolatedumbilicalartery(UA)andvein(U~V)in.vitro. STUDY DESIGN: IntactUA andUVringsweresuspendedin 5 ml organbathscontaining Kreb'ssolution(pH 7.2; 37°C;bubbledwith 2.5% O~+ 8.07% CO2/ balanceN2)for isometrictensionrecording.Meneninm(finalbathconcentration- 1%),centrifugedand filteredmecenium,and mecnoiumwith the lipid fractionremoved(separatedinto < and > 30,000 KD),was addedto the baths. Somevesselswereincubatedin 1% maceniwnfor 30 minonteswhichwas thenwashedout. Concentration-responsocurvesto U46619 were obtained.TheEChoHog of the concentrationwhichevokes50% of the maximal contraction)was dcterndnnd.Analysis:ANOVA(with nunn's test), Significance;p < 0.05. S,o~ ~ . .u. .m. ~1%I RESULTS: IEC_.) UAInl UVIn, ~t29, ~w'ld Control 8.1+0.3(16) 8.0+0.4(11) ~ ~oo] / ~ ; : ~ Maconium1% 7.4+ 0.2" (10) 7.3+0.3"(11) :E =~ Meconium(washed) 7.9+0.4m(8) 7.6+0.4(9) ~ 6o Meconium(filtered) 7.7+0.3* (9) 7.4+0.5*(6) ~,o -6to Lipid Removed ~ o . . . . . . -s5 4o -r.5 -7o < 30.000 KD 7.9+0.4m(8) NotDono .8o U4eSlQnoaM1 > 3D.000 KD 8.0+ 0.2m(61 Not Done * - differentfromcontrol(p< 0.05); m, differentfrommaconium1% (p< 0.05) UA andUVhadno sustainedresponseto mecnoium.Tensiondevelopedby ringsexposedto meconiumwas significantlylessthancontrolat all concentrationsof U46619. Therewas somelossof the efficacyOfmeconiumfollowingcentrifugation/fifirationand washingout. Meconiumwithoutthe lipid fractionhadlessinhibitoryeffect than nativemeconium CONCLUSION: Meconiumdoesnot causesustainedcontractionof UA or UVin-vitro. It does,however,inhibitthe contractileeffect of U46819in both UA and UVsmoothmuscle. TheinhibitoryactivityQfmoconiumappearsto be concentratedin the lipidfraction.
180
LOCALIZATION OF NITRIC OXIDE SYNTHASE ACTIVITY IN VARIOUS COMPARTMENTS OF HUMAN PLACENTA. P.F. Mentox, P.I. Karlx, E.P. Girardix, B.M. Pctrikovsky and B.M. Wilkesx. Depts of Nephrology, Pediatrics and Fetal-Maternal Medicine, North Shore Univ Hospital-Comell Univ Med College, Manhassct, NY OBJECTIVE: We tested the hypothesis that nitric oxide synthas¢ (NOS) activity is differentially distributed in the placenta and preferentially expressed in the vascular compartment. STUDY DESIGN: Tissues from vascular and non-vascular compartments of 12 normal placentas were homogenized and separated into cytosolic and membrane fractions by ultraccntrifugation. NOS activity was measured by conversion of [14C]-arginine to [14C]-citrulline. Activity in wholo placental homogcnat¢ was compared with activity in human umbilical vein endothelial cells (HUVEC), small placental arteries (<1.0 mm diameter) and syncytiotrophoblasts in culture. RESULTS: In whole placenta, HUVECs, and small placental arteries, 80% of the NOS activity was in the membrane fraction with the remaining activity present in the cytosol. In contrast, in cultured syncytiotrophoblasts NOS activity occurred exclusively in the membrane fraction. NADPHdiaphorase staining of whole placenta was found only in the syncytiotrophoblasts. Day O isolated trophoblasts showed little diaphorasc staining and NOS activity. On days I and 3, as syncytiotrophoblasts differentiated, diaphoras¢ staining and NOS activity increased in parallel. CONCLUSIONS: The cellular distribution of nitric oxide synthas¢ activity was different in vascular and non-vascular compartments s!nce acitivity in syncytiotrophoblasts occurred exclusively in the membrane fraction. Nitric oxide synthesis increases as trophoblasts mature. These data suggest a potential role for nitric oxide in platelet adhesion in the intervillous space and regulation of resistance in fetal arterioles.
Y