Validation of Quintero stage III sub-classification for twin-twin transfusion syndrome based on visibility of donor bladder: characteristic differences in pathophysiology and prognosis.

OBJECTIVE: To validate the Quintero stage III subclassification for twin-twin transfusion syndrome (TTTS) based on visibility of the bladder of the donor twin. METHODS: Between July 2002 and August 2006, there were 131 pregnant Japanese women affected by severe TTTS before 26 weeks' gestation, treated with fetoscopic laser surgery at five centers in Japan, whose pregnancies continued beyond 22 weeks. Outcome data were available in all cases and surviving infants were followed up for at least 6 years. This study focused on the Stage III TTTS patients. These were subclassified into Stage III atypical (abnormal Doppler flow with visible donor bladder) and Stage III classical (abnormal Doppler flow with non-visible donor bladder) groups. Perioperative data and postnatal outcomes were compared between the groups. RESULTS: Seven Stage I, 22 Stage II, 82 Stage III and 20 Stage IV pregnancies continued beyond 22 weeks. There was a significantly higher incidence of absent or reversed end-diastolic velocity in the umbilical artery (UA-AREDV) of the donor in Stage III atypical than in Stage III classical patients (83.8% vs. 53.3%, P = 0.004). Stage III atypical cases also had a significantly higher incidence of arterioarterial (AA) anastomoses (72.9% vs. 17.8%, P < 0.001) and intrauterine fetal demise (IUFD) of the donor (43.2% vs. 13.3%, P = 0.002). However, there were no differences in overall survival or in abnormal brain scans of surviving infants. Donors with both UA-AREDV and AA anastomoses had a significantly higher incidence of IUFD compared with the others (53.3%, P < 0.001). CONCLUSIONS: Quintero stage III atypical was characterized by a high incidence of AA anastomoses and UA-AREDV of the donor, resulting in IUFD. Subclassification of Stage III based on visibility of the bladder of the donor twin was adequate for and compatible with differentiating prognosis and pathophysiology.

Detection of a biorhythm of human fetal autonomic nervous activity by a power spectral analysis.

OBJECTIVE: This study describes a 24-hour assay of autonomic nervous system activity in human fetuses with the use of a power spectral analysis of heartbeat intervals. STUDY DESIGN: The study included 6 normal fetuses and 11 growth-restricted fetuses, 3 of which were hypoxemic. Fetal heartbeats were continuously recorded for 24 hours, and a power spectral analysis was performed on the beat interval data. A low frequency domain of beat intervals was identified (0.025-0.125 cycles/beat) and evaluated at regular intervals over the 24-hour period. The cosinor method was used to detect rhythmic fluctuations in the resulting low frequency areas. The same procedures were also performed with the mothers of the fetuses. RESULTS: We observed 2 diurnal rhythms of heartbeat activity (1 with a 12-hour period and another with a 24-hour period) in the normal and growth-restricted fetuses without hypoxemia. In contrast, these rhythms were not observed in the hypoxemic fetuses. In the mothers, only the 24-hour rhythm was observed. CONCLUSION: The 12-hour cycle of autonomic nervous activity is present in normal fetuses.

Compound heterozygous mutations (PHE53/54DEL and HIS373LEU) of the P450c17 gene result in a 17alpha-hydroxylase/17,20-lyase deficient male pseudohermaphrodite with unambiguous external genitalia.

The autosomal recessive disease 17alpha-hydroxylase/17,20-lyase deficiency is characterized by mutation of the P450c17 enzyme, which catalyzes 17alpha-hydroxylation and 17,20-lysis in the steroidogenic pathways. Although 17 mutations of this enzyme have been reported, only a few of them resulted in a completely unambiguous phenotype of female external genitalia in 46, XY individuals. We report here a Japanese patient with a 46,XY karyotype, who showed such a unambiguous female external genitalia. Nucleotide sequencing of the P450c17 gene revealed the patient to be a compound heterozygote carrying two different mutations (PHE53/54DEL in exon 1 and HIS373LEU in exon 6). As these mutations have been previously detected in unrelated Japanese patients, it is confirmed that these mutations accumulate regionally. Since these mutations could be screened by a multiple genotyping method, the method is applicable when 17alpha-hydroxylase/17, 20-lyase deficiency is suspected in Japanese patients.

Power spectral analysis of R-R interval variability before and during the sinusoidal heart rate pattern in fetal lambs.

OBJECTIVE: The appearance of the sinusoidal heart rate pattern found on fetal cardiotocograms has not been fully explained, either physiologically or clinically. In this study we performed power spectral analysis on the sinusoidal heart rate pattern obtained by administration of arginine vasopressin and atropine sulfate to investigate its frequency components in fetal lambs with long-term instrument implantation. STUDY DESIGN: Eleven tests were performed in 4 fetal lambs at 120 to 130 days' gestation. An artificial sinusoidal heart rate pattern was obtained by administration of atropine sulfate and arginine vasopressin in 9 tests. An autoregression model was used to compare the spectral patterns before and during the sinusoidal heart rate pattern. RESULTS: Marked decreases in low-frequency (0.025-0.125 cycles/beat) and high-frequency (0.2-0.5 cycles/beat) areas were observed in the presence of the sinusoidal heart rate pattern. However, there were no significant changes in the very-low-frequency area (0.01-0.025 cycles/beat), which corresponds to the frequency of the sinusoidal heart rate pattern. CONCLUSION: The sinusoidal heart rate pattern may represent a very low-frequency component inherent in fetal heart rate variability that appears when low- and high-frequency components are reduced as a result of strongly suppressed autonomic nervous activity.

Growth pattern of twins of different chorionicity evaluated by sonographic biometry.

OBJECTIVE: To determine differences between growth patterns of monochorionic and dichorionic twins, and of concordant and discordant twins of both chorionicities. METHODS: We studied 70 cases of concordant twins (24 monochorionic, 46 dichorionic) and 45 cases of discordant twins (25 monochorionic, 20 dichorionic). In each case, growth was measured longitudinally by ultrasound biometry and the growth pattern was depicted. RESULTS: There were no differences in incremental growth between concordant monochorionic and dichorionic twins. The growth curve of concordant twins of both chorionicities was almost the same as that of a singleton until 34 weeks' gestation. However, in the discordant twins, the growth of the larger twin matched the growth curve of a singleton or concordant twin, but the growth of the smaller twin gradually decreased to the range of growth restriction. The growth curves for monochorionic discordant twins appeared to be representative of two groups, one of which had onset of discordancy before 24 weeks. CONCLUSION: It is clinically important to determine chorionicity early in twin pregnancies, to calculate the percentage of discordancy between the fetuses, and to examine longitudinal fetal growth curves in each chorionicity.

Changes in surfactant-associated protein mRNA profile in growth-restricted fetal sheep.

To test the hypothesis that chronic placental insufficiency resulting in fetal growth restriction causes an increase in fetal lung surfactant-associated protein (SP) gene expression, we embolized chronically catheterized fetal sheep (n = 6) daily using nonradioactive microspheres in the abdominal aorta for 21 days (between 0.74 and 0.88 of gestation) until fetal arterial oxygen content was reduced by approximately 40-50%. Control animals (n = 7) received saline only. Basal fetal plasma cortisol concentration was monitored. At the end of the experiment, fetal lung tissues were collected, and ratios of tissue levels of SP-A, SP-B, and SP-C mRNA to 18S rRNA were determined by standard Northern blot analysis. Total DNA content of fetal lungs was reduced by 30% in the embolized group compared with control group (P = 0.01). There was a 2.7-fold increase in fetal lung SP-A mRNA (P < 0.05) and a 3.2-fold increase in SP-B mRNA (P < 0.01) in the chronically embolized group compared with those in the control group. SP-A and SP-B mRNA tissue levels were highly correlated with the mean fetal plasma cortisol levels on days 20-21 (r = 0.90, P < 0.01 for SP-A mRNA and r = 0.94, P < 0.01 for SP-B mRNA). SP-C mRNA tissue levels were not significantly affected by placental insufficiency. We conclude that fetal growth restriction due to placental insufficiency is associated with alterations in fetal lung SP, suggesting enhanced lung maturation that was highly dependent on the degree of increase in fetal plasma cortisol levels.

Effect of chronic hypoxemia on the regulation of nitric-oxide synthase in the fetal sheep brain.

We tested the hypothesis that chronic hypoxemia modulates NO production of the fetal brain by altering its gene and protein expression. Chronically instrumented preterm fetal sheep were made hypoxemic by placental embolization for 21 days. Fetal oxygen content was measured to determine the level of hypoxemia. The expression of both eNOS and nNOS proteins and mRNA and enzyme activities of fetal sheep cerebrum were measured and compared between normoxic and hypoxemic animals. Our results show that in utero hypoxemia downregulates both Ca2+ dependent NOS activity and expression of eNOS protein and mRNA in the fetal sheep brain. In contrast, hypoxemia increased nNOS protein and mRNA levels in the cerebrum. This suggests that chronic hypoxemia has an opposing effect on eNOS and nNOS gene regulation. We propose that increased nNOS activity during chronic hypoxemia may excessively stimulate the neurons and contribute to fetal brain injury. On the other hand, downregulation of eNOS activity and expression may compromise the neuroprotective effect of eNOS and, therefore, further exacerbate brain injury.

Chronic hypoxemia selectively down-regulates 11beta-hydroxysteroid dehydrogenase type 2 gene expression in the fetal sheep kidney.

The present study was designed to examine the effects of chronic fetal placental embolization on the expression of 11beta-hydroxysteroid dehydrogenase (11beta-HSD) types 1 and 2, the intracellular enzymes responsible for the metabolism of glucocorticoids. Twelve instrumented fetal sheep were randomly allocated on Day 110 (term = 147 days) to either a control (n = 6) or embolized (n = 6) group. Embolized fetuses received daily injections of nonradioactive microspheres into the abdominal aorta for 21 days to decrease arterial oxygen content by 40-50% of the pre-embolization values. At the end of the experiment, fetal liver and kidney tissues as well as placental cotyledons were collected, and tissue levels of 11beta-HSD mRNA and activity were determined by standard Northern blot analysis and radiometric conversion assay, respectively. There was a 44% reduction (p < 0.01) in the level of renal 11beta-HSD2 mRNA in the embolized group as compared with the control group. Moreover, this reduction in mRNA was carried through to 11beta-HSD2 protein, since there was a corresponding decrease in the level of 11beta-HSD2 activity (4.5+/-0.2 vs. 2.9+/-0.1 pmol/min per milligram protein, p < 0.01). In contrast, levels of both 11beta-HSD1 mRNA and activity in the fetal liver remained unchanged. Moreover, both 11beta-HSD types 1 and 2 mRNA and activity in the placenta were not altered by the fetal placental embolization. In conclusion, chronic hypoxemia selectively inhibits renal 11beta-HSD2 mRNA expression and enzyme activity in the ovine fetus, which may contribute, at least in part, to the mechanisms leading to fetal hypertension.

Fetal sheep endocrine responses to sustained hypoxemic stress after chronic fetal placental embolization.

The purpose of this study was to determine the endocrine and circulatory responses of the ovine fetus, near term, to sustained hypoxemic stress superimposed on chronic hypoxemia. Fetal sheep were chronically embolized (n = 7) for 10 days between 0.84 and 0.91 of gestation via the descending aorta until arterial oxygen content was decreased by approximately 30%. Control animals (n = 8) received saline only. On experimental day 10, both groups were embolized over a 6-h period until fetal arterial pH decreased to approximately 7.00. Regional distribution of lower body blood flows was measured on day 10, before and at the end of acute embolization. On day 10, the chronically embolized group had lower arterial oxygen content (P < 0.05), Po2 (P < 0.01), and placental blood flow (P < 0.05) than controls and higher prostaglandin E2 (PGE2) and norepinephrine plasma concentrations (both P < 0.05). In response to a superimposed sustained hypoxemic stress, there was a twofold greater increase in PGE2 in the chronically embolized group than in the control group (P < 0.05). However, the increase in fetal plasma cortisol in response to superimposed hypoxemic stress was similar in both groups, despite significantly lower adrenocorticotropic hormone and adrenal cortex blood flow responses in the chronically hypoxemic group (both P < 0.05). We conclude that PGE2 response to a sustained superimposed reduction in placental blood flow, leading to metabolic acidosis, is enhanced under conditions of chronic hypoxemia and may play an important role for the maintenance of the fetal cortisol response to an episode of superimposed acute stress.

Fetal heart rate patterns in growth-restricted fetal sheep induced by chronic fetal placental embolization.

OBJECTIVE: Our purpose was to test the hypothesis that chronic placental insufficiency and intrauterine growth restriction in fetal sheep causes a decrease in the number of fetal heart rate accelerations and fetal heart rate variability. STUDY DESIGN: Chronically catheterized fetal sheep were embolized (n = 6) daily with 15 to 50 microns latex microspheres for 21 days between 0.74 and 0.88 of gestation into the abdominal aorta, until fetal arterial oxygen content was decreased by 40% to 50% of the preembolization value. Control animals (n = 6) received saline solution only. Signals from chest electrodes were analyzed on-line with the Sonicaid System 8000 in 2-hour epochs every 6 hours starting at 8 AM over the first 48 hours of hypoxemia and for 2 hours between 8 and 10 AM every other day from day 3 to day 21 of hypoxemia. Umbilical artery Doppler-derived resistance index and fetal plasma catecholamine concentrations were also measured. RESULTS: Embolized fetuses had asymmetric intrauterine growth restriction and became chronically hypoxemic (p < 0.001) with a progressive increase in the umbilical artery resistance index (p < 0.001). During the first 48 hours of hypoxemia the number of accelerations and decelerations and both short- and long-term fetal heart rate variability increased initially, followed by a return to control levels by 20 hours after the onset of embolization. After 21 days of hypoxemia the number of accelerations was significantly reduced by 30% compared with controls (p < 0.05). Both short- and long-term fetal heart rate variability in control fetuses gradually increased with advancing gestational age (p < 0.001 and p < 0.01, respectively), whereas in embolized fetuses the fetal heart rate variability remained unchanged and was 20% lower than that of controls on day 21 (both p < 0.01). CONCLUSION: Intrauterine growth restriction and long-term hypoxemia in fetal sheep are associated with a decrease in short- and long-term fetal heart rate variability, possibly because of a delay in the normal maturational changes of the autonomic control of fetal heart rate.

Chronic fetal placental embolization and hypoxemia cause hypertension and myocardial hypertrophy in fetal sheep.

To examine the cardiovascular effects on the fetus of an elevated umbilical vascular resistance resulting in fetal hypoxemia, we embolized the fetal side of the placenta in pregnant sheep and measured cardiovascular and hormonal changes and cellular growth in fetal heart. Chronically catheterized fetal sheep were embolized (n = 6) for 21 days between 0.74 and 0.88 of gestation into the descending aorta until arterial oxygen content was decreased by 40-50% of the preembolization value. Control animals (n = 6) received saline only. During embolization, fetuses became chronically hypoxemic (P < 0.001) and hypertensive (P < 0.001), with a progressive increase in umbilical artery resistance index (P < 0.001). There was also an increase in fetal plasma norepinephrine throughout the study period (P < 0.05). On day 21 of embolization, fetuses showed asymmetrical growth restriction, increased heart weight (P < 0.01), and increase in right and left ventricular wall thickness (P < 0.05) compared with control animals. The protein-to-DNA ratio, an index of cell size, increased in the right ventricular myocardium in the embolized group (P < 0.001), suggesting myocardial cell hypertrophy. We conclude that, during chronic placental damage leading to fetal hypoxemia with an increase in umbilical artery resistance index, fetuses developed arterial hypertension and asymmetrical growth restriction and that increases in afterload to the heart and plasma norepinephrine likely caused fetal myocardial hypertrophy.

Effects of long-term hypoxemia on pituitary-adrenal function in fetal sheep.

To test the hypothesis that long-term hypoxemia causes premature activation of the fetal pituitary-adrenal function, we embolized the fetal side of the placenta in pregnant sheep and examined the changes in concentrations of immunoreactive adrenocorticotropic hormone (irACTH), cortisol, and prostaglandin E2 (PGE2) in fetal plasma, and levels and localization of proopiomelanocortin (POMC) mRNA in the pars distalis and the pars intermedia of the fetal pituitary. Twelve fetal sheep were studied (6 embolized and 6 control) for 21 days between 0.74 and 0.88 of gestation. Daily injections of nonradiolabeled microspheres were given into the fetal abdominal aorta to decrease fetal arterial oxygen content by 40-50% of the preembolization values. In the embolized group, concentrations of irACTH, PGE2, and cortisol in fetal plasma increased gradually and were significantly (P < 0.05) elevated above those of controls after day 10, day 16, and day 20, respectively. POMC mRNA levels in the pars distalis of the fetal pituitary were not different from those of controls but were significantly reduced in the pars intermedia (P < 0.05). We conclude that levels of POMC mRNA in the pars distalis are unchanged during long-term hypoxemia possibly because of negative feedback effects of elevated cortisol on the pituitary gland. During long-term fetal hypoxemia, there is a differential regulation of POMC mRNA expression in the pars distalis and pars intermedia.

Power spectral analysis for autonomic influences in heart rate and blood pressure variability in fetal lambs.

Variability of R-R intervals and arterial blood pressure signals in chronically instrumented fetal lambs was analyzed by power spectral analysis based on an assumption of maximum entropy. There were four consistent components, very low (VL, 0.01-0.025 cycle/beat), low (L, 0.025-0.125 cycle/beat), middle (M, 0.125-0.2 cycle/beat), and high (H, 0.2-0.5 cycle/beat), in the normal heart rate variability and blood pressure spectra. Integrated peaks in the power spectrum were compared before and after the administration of sympathetic and parasympathetic blockades. beta-Sympathetic blockade reduced the spectral power in the VL and L frequency components. alpha-Sympathetic blockade reduced only the M frequency component in the spectrum of R-R interval variability. Parasympathetic blockade reduced the H and L frequency components in the R-R interval variability spectrum but increased these components in the systolic blood pressure variability spectrum. The results clearly demonstrate the association between fetal autonomic activity and change of power spectrum of heart rate and blood pressure variability.

Fetal placental embolization in the late-gestation ovine fetus: alterations in umbilical blood flow and fetal heart rate patterns.

OBJECTIVE: Our goal was to determine the effect of chronic and acute umbilical-placental embolization on placental hemodynamic and fetal heart rate patterns in relation to fetal oxygenation in the near-term ovine fetus. STUDY DESIGN: Daily fetal placental embolization was performed during 10 days in 9 sheep fetuses until fetal arterial oxygen content decreased by approximately 30%. Nine control fetuses received saline solution. Mean and pulsatile umbilical blood flow, perfusion pressure, placental vascular resistance, fundamental impedance, pressure pulsatility index, and umbilical artery resistance index corrected to a fetal heart rate of 160 beats/min were measured. On day 10 both groups were acutely embolized until fetal arterial pH decreased to approximately 7.00. Fetal heart rate was measured with the Sonicaid System 8000 (Oxford Sonicaid, Oxford, United Kingdom). RESULTS: Chronic fetal placental embolization was associated with a progressive reduction in umbilical blood flow (p < 0.00001) and fetal arterial oxygen content (p < 0.001) whereas fetal heart rate patterns remained unaltered. A chronic increase in umbilical artery resistance index corrected to a fetal heart rate of 160 beats/min could be entirely explained only if the changes in umbilical artery pressure pulsatility index and the fundamental impedance were taken into account, in addition to the changes observed in placental vascular resistance. During acute embolization leading to a 50% reduction in umbilical blood flow (p < 0.0002) and a three times increase in placental vascular resistance (p < 0.0001), the most consistent change in fetal heart rate patterns related to progressive metabolic acidosis was an 84% decrease in absolute acceleration frequency (p < 0.0001) whereas short-term fetal heart rate variability remained unaltered. CONCLUSION: Changes in umbilical artery resistance index induced by chronic umbilical-placental embolization resulting in fetal hypoxemia occurred before any changes in fetal heart rate patterns were detectable. A decrease in the absolute acceleration frequency was the only component of fetal heart rate patterns related to progressive metabolic acidosis in the near-term ovine fetus.

Increased fetal plasma prostaglandin E2 concentrations during fetal placental embolization in pregnant sheep.

OBJECTIVE: The purpose of this study was to examine the effect of chronic fetal placental embolization on fetal plasma prostaglandin E2 concentrations. STUDY DESIGN: Fourteen pregnant sheep were studied (seven embolized and seven controls) for 10 days between 0.84 and 0.91 of gestation. Daily injections of nonradioactive microspheres were made to decrease fetal arterial oxygen content by 30% to 35% of preembolization values. RESULTS: In response to repeated embolization, fetal plasma prostaglandin E2 concentrations increased significantly on day 1, declined to near control levels on days 2 to 6, but were significantly elevated again after day 7. Maternal prostaglandin E2 levels remained unchanged throughout the study. Fetal plasma prostaglandin E2 levels increased significantly with decreasing fetal oxygenation when fetal arterial oxygen content was < 2.0 mmol/L. CONCLUSION: We conclude that there is increased production of prostaglandin E2 by the placenta during progressive fetal hypoxemia induced by fetal placental embolization. We speculate that the progressive increase in prostaglandin E2 may be an important hormonal adaptive mechanism to maintain fetal homeostasis during the development of placental insufficiency.

Production of complete heart block and utero cardiac pacing in fetal lambs.

We produced a complete heart block in 5 fetal lambs at 120 to 135 days of gestation by injecting 10% formaldehyde into the His bundle. We obtained acute experimental data in 5 fetuses and chronic experimental data in 2 fetuses. Immediately after production of the complete heart block, cardiac output was acutely reduced to 43 +/- 9% when compared with the cardiac output at sinus rhythm. It was possible to restore the cardiac output to 80 +/- 7% of the preoperative levels by atrioventricular pacing at the same rate as had been present before the heart block was produced. When the heart rate decreased below 100 beats per minute, further increase of stroke volume did not occur, and output was markedly diminished. This model allows us to study the pathophysiological response of a complete heart block, and offers basic information useful for understanding fetal cardiac functioning.

Diagnostic use of cordocentesis in twin pregnancy.

We performed percutaneous umbilical blood sampling, under ultrasound guidance, in both twins of 11 twin pregnancies in whom twin-to-twin transfusion syndrome was suspected. Chorionicity and the presence of placental vascular anastomoses were assessed postnatally. The hemoglobin concentration, hematocrit and other biochemical variables in fetal blood were evaluated in both twins. The mean hemoglobin difference between the large and small twins was 4.8 g/dl (range 1.8-8.1 g/dl) in 5 monochorionic discordant twin pairs, and 1.2 g/dl in a monochorionic twin pair without discordancy. Hemoglobin values did not differ between dichorionic twins. The mean hematocrit difference between monochorionic discordant twins was 18.3%. In dichorionic discordant twins, the inter-twin hematocrit difference was very small. Total protein and albumin were normal in all twins.

Prenatal diagnosis of congenital cystic adenomatoid malformation of the lung by fetal lung biopsy.

A case of type III congenital cystic adenomatoid malformation of the lung was successfully diagnosed prenatally by fetal lung biopsy. We performed this procedure at 22 weeks of gestation, using a biopsy gun system under ultrasound guidance. The pregnancy was undisturbed by the procedure but as the condition was incompatible with life, an abortion was performed. The diagnosis was confirmed at post-mortem examination. Fetal lung biopsy appears to be a useful method for prenatal diagnosis of fetal lung disorders.

Umbilical venous pressure and Doppler flow pattern of inferior vena cava in the fetus.

Umbilical venous blood pressure (UVP) was measured at fetal blood sampling using cordocentesis in 113 fetuses. Among these, both inferior vena cava (IVC) Doppler flow pattern, when the preload index was set as a parameter of reversed flow, and UVP were measured in 50 fetuses. We evaluated whether direct measurement of UVP, representing fetal central venous pressure, could be replaced by an assessment of reversed IVC blood flow. Normal mean UVP was 4.89 +/- 3.22 mm Hg, whereas no correlation was observed between UVP and gestational week. When the extent of reversed blood flow from the right atrium to the IVC was expressed as a preload index (PLI), there was a significant correlation between the index and UVP (r = 0.463; P < 0.001). When the abnormal range of PLI was set at 0.45, the sensitivity of an estimate of abnormal UVP (more than 1.5 SD plus the mean) was 0.54 and the specificity was 0.89. Measurement of UVP is necessary for the prediction of fetal cardiac function. PLI can be substituted for the UVP measurement. However, PLI should be supplemented with other parameters for an accurate fetal cardiac function evaluation.

Funipuncture for evaluation of hematologic and coagulation indices in the surviving twin following co-twin's death.

OBJECTIVE: To examine the changes in hematologic and coagulation indices in the surviving twin when the co-twin dies because of the twin-twin transfusion syndrome. METHODS: Fetal blood was obtained by funipuncture in seven surviving twins upon the death of their co-twins. Five of them were monochorionic. In one case at 32 weeks' gestation, two repeated funipunctures were done in both twins before and in the surviving twin after the death of the co-twin. Fetal blood was examined for blood coagulation factors as well as complete blood counts. RESULTS: Although coagulation factors were not abnormal, three of the five monochorionic surviving twins had cerebral abnormalities postnatally. The fetal blood profile revealed anemia in the surviving twin, especially in the cases in which funipunctures were performed within 24 hours after the co-twin's death. This demonstrates that acute anemia in the surviving twin was induced by hemorrhage from the larger to the smaller twin at the time of death. CONCLUSION: Following the death of one twin, morbidity in the surviving twin can be produced by hypotensive ischemia of the brain due to hemorrhage through placental vascular anastomoses.