Tumor necrosis factor-alpha in the placenta is not elevated in pre-eclamptic patients despite its elevation in peripheral blood.

PROBLEM: Tumor necrosis factor-alpha (TNF-alpha) is present in human placental and uterine cells at the early and late stages of gestation and promotes the regulation of trophoblast growth and invasion. We evaluated whether TNF-alpha levels in the placenta and blood of pre-eclamptic women differed from those with normal pregnancies. METHOD OF STUDY: The subjects were 39 pregnant women carrying single fetuses (21 normal-pregnant and 18 pre-eclamptic patients). Their average gestational age at entry was 38-39 weeks. Peripheral blood was collected before the onset of labor and separated serum was stored at -20 degrees C. A tissue segment of the placenta was cut and frozen in liquid nitrogen immediately after delivery at -80 degrees C. The frozen placental tissue was added to phosphate-buffered saline. The tissue was fully homogenized and centrifuged. Separated supernatant was stored at -80 degrees C. TNF-alpha levels in separated serum and TNF-alpha and total protein (TP) levels in separated supernatant were measured. The presence of TNF-alpha in the placenta was evaluated by immunohistochemistry in five pre-eclamptic and five normal-pregnant patients. RESULTS: Serum TNF-alpha levels were higher in pre-eclampsia than in normal pregnancies. However, TNF-alpha/TP levels in the placenta did not differ significantly between the two groups. As for TNF-alpha immunostaining of trophoblastic cells in the placenta, it was weak in three and moderate in two of the normal pregnancies, while it was absent in two, weak in one, and moderate in two in the pre-eclampsia group. CONCLUSIONS: We demonstrated no significant increase in TNF-alpha/TP levels in the placenta in pre-eclampsia despite a significant increase in serum TNF-alpha levels. There was no strong immunostaining for TNF-alpha detected by immunohistochemistry in the pre-eclampsia group. These findings suggest that TNF-alpha in the placenta is not a key cytokine to interfere with normal trophoblast invasion into the myometrium in pre-eclampsia, and that sources other than the placenta may contribute to the elevated levels of TNF-alpha found in the circulation of pre-eclamptic patients.

Macrophage colony-stimulating factor levels in amniotic fluid before and after the onset of labor do not differ in normal pregnancies.

PROBLEM: Macrophage colony-stimulating factor (M-CSF) promotes placental growth and maintenance. M-CSF also regulates trophoblast invasion into the placental bed. We evaluated whether M-CSF levels in amniotic fluid during labor contributing to subsequent delivery differed from those before the onset of labor in normal pregnancies. METHOD OF STUDY: This study enrolled 48 Japanese women experiencing normal pregnancies with single fetuses who had no infection. Of these pregnancies, 24 were women during labor: 22 led to subsequent term delivery (labors); two had premature delivery. The other 24 were women without labor underwent cesarean section (controls). These two groups (22 labors and 24 controls) were compared. The average gestational age at entry was 38 weeks of gestation. The women's ages and gestational ages did not differ significantly between the two groups. Amniotic fluid was collected and the M-CSF levels were compared between two groups. The M-CSF level was determined by the sandwich enzyme-linked immunosorbent assay (ELISA) method. RESULTS: The levels of M-CSF in amniotic fluid did not differ significantly between the women during labor and those without labor. CONCLUSIONS: M-CSF in amniotic fluid may not contribute to the onset of labor in term pregnancy and/or labor resulting in subsequent delivery may not induce the production and secretion of M-CSF into amniotic cavity.

Elevation of granulocyte-macrophage colony-stimulating factor in the placenta and blood in preeclampsia.

OBJECTIVE: This study investigated whether granulocyte-macrophage colony-stimulating factor (GM-CSF) levels in the placenta and blood in preeclampsia differed from those in normal pregnancies. Macrophage colony-stimulating factor (M-CSF) levels in the placenta were also measured. STUDY DESIGN: The subjects were 44 pregnant women carrying single fetuses, of whom 22 were women with normal pregnancies and 22 were women with preeclampsia. Their average gestational age at entry was 37 to 38 weeks of gestation. Peripheral blood was collected before the onset of labor. Separated serum was obtained after centrifugation and stored at -20 degrees C. A tissue segment of the placenta was cut immediately after delivery. The frozen placental tissue was placed into a plastic tube containing phosphate-buffered saline solution. The tissue was fully homogenized and then centrifuged. Separated supernatant was frozen at -80 degrees C for subsequent determination. GM-CSF levels in separated serum were measured, and GM-CSF, M-CSF, and total protein (TP) levels in separated supernatant were also measured. RESULTS: Not only GM-CSF levels in blood but also GM-CSF/TP levels in the placenta were significantly higher (P<.05) in preeclampsia than in normal pregnancies. Similar results were obtained for M-CSF/TP levels in the placenta. CONCLUSIONS: We demonstrated a significant increase in placenta levels of GM-CSF/TP in preeclampsia. Elevated GM-CSF in the placenta may be related to immunologic abnormalities contributing to the etiology of preeclampsia.

Elevation of serum macrophage colony-stimulating factor before the clinical manifestations of preeclampsia.

OBJECTIVE: The purpose of this study was to determine whether the increase in serum macrophage colony-stimulating factor (M-CSF) levels preceded the onset of preeclampsia. STUDY DESIGN: We selected 146 women, of whom 36 were nonpregnant women participating in the preliminary study and 110 were normotensive pregnant women at risk for preeclampsia who were carrying single fetuses at about 18 weeks of gestation. The blood was collected and serum was stored at -20 degrees C until assay. Sixteen women had preeclampsia develop at a later stage of pregnancy (preeclamptics), whereas 89 women continued to have normotensive pregnancies until delivery. Thirty-five of the 89 women with normotensive pregnancy who were matched for age and parity were selected to form a control group (controls). Serum M-CSF levels were determined by the sandwich enzyme-linking immunosorbent assay method with use of three antibodies. RESULTS: Serum level of M-CSF was 1295 U/mL (median) in preeclamptics and 957 U/mL in controls. Serum M-CSF levels were significantly higher (P<.0001) in preeclamptics than in controls. CONCLUSION: The increase in serum M-CSF levels markedly precedes the development of clinical manifestations of preeclampsia. Elevation of serum M-CSF supports M-CSF elevation in the placenta. This elevation at 18 weeks of gestation may be related to placental hypoxia, which is considered the cause of preeclampsia.

Serum levels of macrophage colony-stimulating factor after cesarean section, vaginal delivery, or laparotomy in normal pregnant women and gynecologic patients.

BACKGROUND: Macrophage colony-stimulating factor (M-CSF) is located in villous cells lining the vessels in the placenta in the third trimester and has been implicated in placental growth and development. Macrophage colony-stimulating factor levels in peripheral blood increased significantly with progression of pregnancy in uncomplicated pregnancies. The serum levels of M-CSF appear to be altered after laparotomy in normal pregnant women and nonpregnant gynecologic patients. Thus, the present study examined changes in serum levels of M-CSF before and after laparotomy and compared these findings between the two groups. Macrophage colony-stimulating factor levels before and after vaginal delivery were also examined. METHODS: Peripheral blood was collected before, 1 day, and 10 days after laparotomy or vaginal delivery from 38 subjects, of whom 14 were normal pregnant women who underwent cesarean section (group 1), 12 were gynecologic patients (group 2), and 12 were normal pregnant women who delivered vaginally (group 3). The M-CSF level was determined by the sandwich ELISA method using three antibodies. RESULTS: In all groups, the serum levels of M-CSF increased significantly 1 day after laparotomy or vaginal delivery, but then decreased significantly after 10 days. The net increase 1 day after laparotomy was significantly lower in group 1 than in group 2. Before and 1 day after laparotomy, the M-CSF levels were significantly higher in group 1 than in group 2, but not 10 days after laparotomy. Changes in M-CSF levels in group 3 were relatively similar to those in group 1. CONCLUSIONS: Serum levels of M-CSF were significantly higher in groups 1 and 3 than in group 2, before laparotomy or vaginal delivery. The M-CSF level increased moderately 1 day after cesarean or vaginal delivery, and it increased remarkably after gynecologic laparotomy. The increases in M-CSF levels postlaparotomy may occur via different mechanisms between groups 1 and 2. Placental removal and termination of pregnancy might contribute to the decrease in M-CSF levels, leading to only a moderate increase in M-CSF levels 1 day after laparotomy in group 1.

Successful conception following Fallopian tube recanalization in infertile patients with a unilateral proximally occluded tube and a contralateral patent tube.

BACKGROUND: There are many published case reports of successful conception following transcervical Fallopian tube recanalization (T-FTR) in patients with bilateral proximally occluded Fallopian tubes. However, no serial trials have been published with respect to successful conception following unilateral tubal recanalization in infertile patients with a unilateral proximally occluded tube and a contralateral patent tube. This study was designated to analyse the success rate of T-FTR and the pregnancy rate due to natural fertilization in the lumen of the recanalized tube in these patients. METHODS: We have encountered only 11 patients with this abnormality in our department in the past 10 years. T-FTR with fluoroscopic guidance was performed in these patients, confirmed by at least two hysterosalpingographies to exclude tubal spasm. The uterine catheter devised by us was used during the procedure. RESULTS: All 11 Fallopian tubes were successfully opened by T-FTR. In the six patients who conceived, a preovulatory follicle was demonstrated on the side of the cannulated tube during the conception. The success rate of recanalization, the pregnancy rate due to fertilization in the lumen of the recanalized tube and the successful delivery rate were 100, 55 and 36% respectively. CONCLUSIONS: Our findings suggest that a functional and/or organic disorder in the patent tube resulted in infertility in patients with unilateral proximal tubal obstruction. Our results further show that recanalization of occluded tubes is an effective treatment. Thus, recognition of successful conception following T-FTR in these patients will be beneficial to our clinical approach to this infertile condition.

Characterization of five marker levels of the hemostatic system and endothelial status in normotensive pregnancy and pre-eclampsia.

OBJECTIVES: Pre-eclampsia is associated with changes in the hemostatic system and endothelial status. Urinary 11-dehydrothromboxane B2/creatinine (11-DTXB2/Cr) is a marker for platelet activation and vascular constriction, thrombin-antithrombin complex (TAT) for thrombin formation, serum thrombomodulin (TM) for endothelial damage, and beta-thromboglobulin (beta-TG) and platelet factor 4 (PF-4) for platelet activation and releasing reaction. The present study attempted to evaluate these five markers in normotensive pregnancy and pre-eclampsia. METHODS: These five markers were simultaneously measured in urine and blood samples from 25 women who were not pregnant (group 1, controls), 31 women with normotensive pregnancy (group 2, second controls), 22 women with mild pre-eclampsia (group 3), and 21 women with severe pre-eclampsia (group 4). The average gestational age was 36 wk. RESULTS: The 11-DTXB2/Cr, TAT, and beta-TG levels were significantly higher (P < 0.01) in groups 2, 3, and 4 than in group 1. The TM and beta-TG levels were significantly higher (P < 0.05) in group 3 than in group 2. The TM, beta-TG, and PF-4 levels were increased significantly (P < 0.05-0.01) in group 4 compared to those in groups 1, 2, and 3. CONCLUSION: Platelet aggregation, vascular constriction, and thrombin formation (detected by 11-DTXB2/Cr and TAT) may be markedly enhanced even in group 2, but further enhancement may be relatively slight in groups 3 and 4. In contrast, endothelial damage (determined by TM) and platelet release of PF-4 may not increase significantly in group 2, but they may increase in group 4. Platelet-release of beta-TG may be enhanced in groups 2, 3, and 4. Endothelial damage and platelet-releasing reaction (detected by PF-4 and beta-TG) may be significantly more enhanced in group 4 than in group 3.

Changes in blood macrophage colony-stimulating factor levels after cesarean section in normotensive pregnancy and preeclampsia.

BACKGROUND: Macrophage colony-stimulating factor (M-CSF) stimulates the proliferation and differentiation of placental trophoblasts and may regulate trophoblast invasion into the placental bed. M-CSF levels in peripheral blood show a significant increase in preeclampsia. Thus, the present study examined changes in blood levels of M-CSF before and after cesarean section and compared them between normotensive and preeclamptic pregnant women. METHODS: Peripheral blood was collected before, 1 day after, and 10 days after cesarean section from 27 women, 12 of whom were preeclamptic pregnant patients with a mean blood pressure of 162/98 mm Hg and 15 were age- and gestational age-matched normotensive pregnant women (normotensive control subjects). Peripheral blood was also collected once from 15 age-matched healthy, normal cycling women (nonpregnant control subjects). M-CSF level was determined by the sandwich enzyme-linked immunosorbent assay (ELISA) method using 3 antibodies. RESULTS: In normotensive and preeclamptic pregnancies, the M-CSF levels increased significantly (P < 0.01) 1 day after surgery but then decreased significantly (P < 0.01) at 10 days after surgery. Before and 1 day after surgery, the M-CSF levels were significantly higher (P < 0.01) in preeclamptic patients than in normotensive control subjects, but not at 10 days after surgery. CONCLUSIONS: The blood M-CSF levels were significantly higher in preeclampsia than in normotensive pregnancies, before cesarean section. The M-CSF levels in the circulation at 1 day after surgery increased significantly. The increase was about 270 U/mL net and at similar levels in 2 groups. Thus, increases in M-CSF levels after cesarean section may occur via similar mechanisms in normotensive and preeclamptic pregnancies. The M-CSF level in normotensive pregnancies and preeclampsia decreased and returned to the normal level at 10 days after cesarean section.

Elevated levels of serum macrophage colony-stimulating factor in normotensive pregnancies complicated by intrauterine fetal growth restriction.

OBJECTIVE: Macrophage colony-stimulating factor (M-CSF) is considered an essential cytokine for placental growth and maintenance. M-CSF also may regulate trophoblast invasion into the placental bed. The aim of the present study was to evaluate whether serum M-CSF levels were altered in normotensive pregnancies complicated by intrauterine growth restriction (IUGR) arising from unknown factors. Plasma thrombin-antithrombin complex (TAT) levels and the pulsatility index (PI) values also were measured. PATIENTS AND METHODS: This study enrolled 47 Japanese women experiencing normotensive pregnancies with single fetuses. Of these pregnancies, 20 were complicated by IUGR arising from unknown factors; these women later delivered small-for-gestational-age (SGA) infants. The other 27 women later delivered appropriate-for-gestational-age infants (controls). The women's ages and gestational ages did not differ significantly between the two groups. Maternal peripheral blood was collected, and the levels of serum M-CSF and plasma TAT were compared between two groups. The M-CSF level was determined by the sandwich enzyme-linked immunosorbent assay method and the TAT level by the enzyme immunoassay method. The PI value for the middle cerebral artery of the fetuses was calculated. RESULTS: Serum levels of M-CSF were significantly higher (p < 0.005) in pregnancies complicated by IUGR that produced SGA infants than in controls. Plasma levels of TAT also were significantly higher (p < 0.02) in pregnancies that produced SGA infants than in controls. The PI values were significantly lower (p < 0.05) in pregnancies that produced SGA infants than in controls. CONCLUSIONS: This study demonstrated significant increases in serum M-CSF levels in women with normotensive pregnancies complicated by IUGR arising from unknown factors who later delivered SGA infants. To the best of our knowledge, this is the first such report. Elevated serum M-CSF levels may be related to placental hypoxia leading to pregnancies complicated by IUGR.

The levels of five markers of hemostasis and endothelial status at different stages of normotensive pregnancy.

BACKGROUND: Urinary 11-dehydrothromboxane B(2)/creatinine (11-DTXB(2)/Cr) is a marker for in vivo platelet activation and vascular constriction, blood thrombomodulin (TM) for endothelial damage and associated thrombosis, thrombin-antithrombin complex (TAT) for thrombin formation, and beta-thromboglobulin (beta-TG) and platelet factor 4 (PF-4) for in vivo platelet activation and releasing reaction. Little is known about the quantitative relationship among them during pregnancy. The present study investigated levels of five markers at different stages of normotensive pregnancy. METHODS: Subjects were 17 healthy non-pregnant women (Group 1, control) and 67 women carrying single fetuses in normotensive pregnancy. Of the pregnant women, 17 were in the 20th week of gestation (Group 2), 20 were in their 30th week (Group 3), and 30 were in their 36th week (Group 4). Urinary and circulating blood levels of 11-DTXB(2)/Cr, TM, TAT, beta-TG, and PF-4 were measured simultaneously. RESULTS: The 11-DTXB(2)/Cr and TAT levels showed elevated values at the 20th and 30th weeks of gestation, and markedly elevated values at the 36th week, whereas the TM level remained constant throughout pregnancy. The beta-TG and PF-4 levels maintained stable values at the 20th week, but showed elevated values at the 30th and 36th weeks. CONCLUSIONS: Platelet aggregation, vascular constriction, and thrombin formation (detected by 11-DTXB(2)/Cr and TAT) appear to be enhanced as early as the 20th week of gestation, continuously enhanced by the 30th week, and markedly enhanced by the 36th week. Platelet activation and releasing reaction (determined by beta-TG and PF-4) gradually enhanced from the 30th to 36th weeks. In contrast, endothelial damage and associated thrombosis (detected by TM) were minimal throughout pregnancy. Investigating these markers of hemostasis and endothelial function in normotensive pregnancy may provide insights into related disease states.

Increased levels of macrophage colony-stimulating factor in the placenta and blood in preeclampsia.

PROBLEM: Macrophage colony-stimulating factor (M-CSF) is considered an essential cytokine for placental growth and maintenance. We evaluated whether M-CSF levels in the placenta and blood in preeclampsia differed from those in normal pregnancies. METHOD OF STUDY: The subjects were 37 pregnant women carrying single fetuses, of whom 19 were women with normal pregnancies and 18 were women with preeclampsia. Their average gestational age at entry was 38 weeks of gestation. Blood was collected before the onset of labor, and separated serum was obtained after centrifugation. A tissue segment of the placenta was cut immediately after delivery. The frozen placental tissue was placed in a plastic tube containing phosphate-buffered saline. The tissue was fully homogenized and then centrifuged. Separated supernatant was used for subsequent determination. M-CSF levels in separated serum were measured, and M-CSF and total protein (TP) levels in separated supernatant were also measured. RESULTS: Both M-CSF/TP levels in the placenta and M-CSF levels in blood were significantly higher (P < 0.05-0.01) in preeclampsia than in normal pregnancies. CONCLUSIONS: This is the first report concerning high placenta levels of M-CSF/ TP in preeclampsia. Increased M-CSF in the placenta supports the hypothesis that immunological abnormalities contribute to the etiology of preeclampsia.

Changes in urinary excretion of six biochemical parameters in normotensive pregnancy and preeclampsia.

We evaluated renal functions by urinary biochemical parameters in normotensive pregnancy and preeclampsia. The parameters are expected to be altered resulting from different abnormalities of renal glomeruli and tubules. We chose N-acetyl-beta-d-glucosaminidase (NAG), beta2-microglobulin (beta2MG), total protein (TP), albumin (Alb), urea nitrogen (UN), uric acid (UA), and creatinine (Cr). Urinary excretion of these biochemical parameter concentrations (relative to Cr) was measured simultaneously in first morning fasting urine samples from 27 healthy nonpregnant women (group 1), 32 women with normotensive pregnancies (group 2), and 26 women with preeclampsia (group 3). The average gestational age at entry was 36 weeks. Serum UN and serum UA also were measured. All the ratios were significantly higher in group 2 than in group 1. The NAG-to-Cr, TP-to-Cr, and Alb-to-Cr ratios were significantly higher in group 3 than in group 2. In contrast, the UN-to-Cr and UA-to-Cr ratios were significantly lower in group 3 than in group 2. The percent increase in the beta2MG-to-Cr ratio in group 2 relative to that in group 1 was the highest, followed by percent increases in the NAG-to-Cr, TP-to-Cr, Alb-to-Cr, UA-to-Cr, and UN-to-Cr ratios. In contrast, the percent increase in the Alb-to-Cr ratio in group 3 relative to that in group 2 was the highest, followed by percent increases in the TP-to-Cr, NAG-to-Cr, beta2MG-to-Cr, UA-to-Cr, and UN-to-Cr ratios. The percent increases in the NAG-to-Cr and beta2MG-to-Cr ratios rose markedly in normotensive pregnancy, whereas percent increases of the Alb-to-Cr and TP-to-Cr ratios were far greater in preeclampsia than in normotensive pregnancy. Renal tubular damage and reabsorption dysfunction may be impaired markedly even in normotensive pregnancy, and further deterioration in reabsorption dysfunction may be slight in preeclampsia. Renal glomerular permeability of TP and Alb may be enhanced in normotensive pregnancy and markedly enhanced in preeclampsia.