The increase in fetal hemoglobin synthesis in the fetal lamb during hyperglycemic hypoxemia

To determine whether fetal hypoxemia induced by hyperglycemia has any effect on the proportions of fetal and adult hemoglobin synthesized during fetal development, hemoglobin synthesis was determined after a period of hyperglycemic hypoxemia in the fetal lamb. These experiments were carried out at a time in gestation during the switch from fetal to adult hemoglobin. Twelve catheterized fetal sheep were included in this study. Seven were made hyperglycemic by glucose infusions during 5 days (group I) and five were control animals (group II) that received saline solution infusions (0.45 gm/dl). In group I, glycemia increased from 14.7 +/- 5.0 to 54.6 +/- 16.4 mg/dl (p less than 0.001), whereas oxygen content decreased from 8.5 +/- 1.7 to 6.4 +/- 2.2 ml/dl (p less than 0.01). Red blood cells obtained before and after 5 days of glucose or saline solution infusions were incubated in an amino acid mixture containing 14C-leucine. The hemoglobins were then subjected to polypeptide chain elution with carboxymethyl cellulose chromatography. The amount of fetal hemoglobin synthesized was determined by the ratio of radioactive gamma-chain to total of radioactive non-alpha-chains. The data demonstrated that the hyperglycemic fetus synthesizes more fetal hemoglobin than expected for the period of fetal development (78.0% +/- 10.9% versus 59.8% +/- 11.3%, p less than 0.02).     

Changes in the ST waveform of the fetal lamb electrocardiogram with hypoxemia

The ST waveform of the fetal electrocardiogram (ECG) was examined in 10 chronically instrumented fetal lambs from 115 days to term. Averaged ST waveforms were plotted at 5-minute intervals in six fetuses for 2 to 22 days. No diurnal or other rhythms were seen. To correct for changes in signal gain the amplitude of the T wave was measured relative to the amplitude of the QRS complex. The T/QRS ratio was normally less than 0.30. Persistently elevated ST waveforms with a T/QRS range 0.32 to 0.65 preceded fetal death by some days in three fetuses and were associated with anemia and/or hypotension in a further three. In these animals hypoxia produced a further rise in the ST waveform (mean T/QRS, from 0.48 to 0.81) and all died during labor. In lambs with a normal ST waveform there were differences in the response to hypoxia over 1 hour for a similar fall in Po₂. In eight experiments the ST segment and T wave increased (mean T/QRS, from 0.17 to 0.59) and promptly reverted to normal with normoxia. There was a significant rise in mean arterial blood pressure, plasma lactate, and glucose and a fall in pH. In four experiments there was little change (mean T/QRS, from 0.19 to 0.25), with a small rise in plasma lactate suggesting that these lambs were able to maintain aerobic myocardial metabolism. Overall there was a strong correlation of the T/QRS ratio to the rate of rise of lactate. These findings complement previous results in the acute fetal lamb preparation and suggest that ST waveform elevation expressed as the T/QRS ratio identifies a change to anaerobic myocardial metabolism.     

Umbilical artery flow velocity waveforms during acute hypoxemia and the relationship with hemodynamic changes in the fetal lamb

The contributions of the variables of the fetal circulation to changes in the pulsatility index of the umbilical artery flow velocity waveform have not been assessed. Acute fetal hypoxemia was induced by 60 to 90 seconds of total occlusion of the maternal common internal iliac artery in six sheep. Mean fetal PO2 levels decreased from 26.0 to 18.3 mm Hg (p less than 0.01) after occlusion of uterine blood flow. Fetal heart rate decreased from 188 to 121 beats per minute at the end of occlusion (p less than 0.05). Placental vascular resistance did not change during the heart rate deceleration. The pulsatility index increased from 0.86 during the control period to 1.27 at the end of occlusion (p less than 0.05). After fetal parasympathetic blockade with atropine, fetal heart rate and placental vascular resistance did not change during occlusion. The pulsatility index did not change during occlusion after parasympathetic blockade. It is concluded that the changes in the umbilical artery pulsatility index during late decelerations in the fetal heart rate pattern appear to be primarily associated with changes in fetal heart rate and bear no relationship with placental vascular resistance.     

Effects of thyrotropin-releasing hormone in the fetal lamb

Thyrotropin-releasing hormone causes neurobehavioral arousal and stimulates breathing in adult, newborn, and preterm experimental animals. Its effects on behavioral state, breathing, blood pressure, and heart rate were studied in the chronically instrumented late term fetal lamb. Fetal intravenous administration of thyrotropin-releasing hormone resulted in behavioral arousal with electrocortical desynchronization, increased body and eye movements, rapid and deep breathing movements, and a transient bradycardia followed by prolonged tachycardia, associated with an increase in both systolic and diastolic blood pressure. The effects were similar following intracisternal administration of thyrotropin-releasing hormone. The effects of thyrotropin-releasing hormone on behavior, but not breathing, was abolished in the presence of muscarinic blockade. Thyrotropin-releasing hormone may play a role in the modulation of central regulation of cardiovascular, respiratory, and behavioral activity in the fetus.     

Effects of naloxone on fetal circulatory responses to hypoxemia

The effects of opiate receptor antagonism on the fetal cardiovascular response to hypoxemia were examined by means of the radionuclide-labeled microsphere technique. Heart rate, blood pressure, and cardiac output were measured during baseline perods, during hypoxemia, and before and after infusion of either naloxone (1 mg/kg) or an equivalent volume of 0.9% saline solution. Seventeen fetal sheep were subjected to maternal hypoxemia by allowing the ewes to breathe 10% oxygen (3% carbon dioxide, 87% nitrogen). The fetuses responded with bradycardia (p < 0.002 compared with control), increased blood pressure (p < 0.002 compared with control), and no significant change in combined ventricular output or placental blood flow. After naloxone, the bradycardia increased by 10% (p < 0.001), and both combined ventricular outout and placental blood flow fell by 20% (p < 0.01 and p < 0.01, respectively). The fetal bradycardic response to naloxone was reversible with atropine. In fetuses with normal oxygenation of the blood (normoxemic), naloxone had no significant effect on heart rate and blood pressure. These data indicate that endogenous opiates (e.g., endorphin and enkephalin) are important in regulating the fetal circulation during hypoxia, and that the effects of opiate receptor antagonism may be mediated through the autonomic nervous system.     

Effect of acute hypoxemia and respiratory acidosis on the fetal heart rate in monkeys

The fetal heart rate (FHR) response to acute nonacidemic hypoxemia and to acute respiratory acidosis was studied in unanesthetized pregnant monkeys by means of chronically implanted catheters and electrodes. Average FHR and short- and long-term FHR variability were analyzed separately. The FHR response to either hypoxemia or hypercapnia consisted of slowing and increased variability. The slowing was more consistent with hypercapnia than with hypoxemia. Long-term FHR variability increased with both test stresses. The increase in short-term FHR variability was much greater in response to hypoxemia than to hypercapnia. Both the FHR slowing and the increases in variability in response to the test stresses were reduced by prior administration of atropine to the fetus.     

Effects of clonidine on breathing movements and electrocortical activity in the fetal lamb

Clonidine is a recommended antihypertensive for use during pregnancy, although little is known of its fetal effects. This study examines the effects of clonidine on breathing and sleep-state cycling in fetal lambs. Clonidine was infused into a fetal lateral ventricle for up to 24 hours at 128 to 135 days' gestation. Control infusions of artificial cerebrospinal fluid had no effect. Clonidine infusion significantly reduced the incidence and episode duration of fetal breathing for the duration of the infusion period. Cycling of electrocortical activity became irregular and rapid, and the incidence of high-voltage electrocortical activity (equivalent to quiet sleep) was reduced. Fetal heart rate decreased but arterial pressure was unaffected. After infusion the breathing incidence and episode duration both increased significantly compared with control, with continuous high-amplitude breathing for several hours, whereas the incidence of high-voltage electrocortical activity remained low. Because lung development is promoted by fetal breathing, long-term use of clonidine during pregnancy could slow lung development by reducing fetal breathing activity.     

Accuracy of echocardiography measurements in the fetal lamb

Echocardiography is becoming an important noninvasive method to evaluate the heart of the human fetus. Validation of this technique, however, is lacking. To establish the correlation of M-mode and two-dimensional echocardiography with direct invasive methods, the size and volume of the heart of 10 fetal lambs were determined. Both M-mode echocardiography as well as the two-dimensional method were found to correlate well with direct measurements. As determined by all three modalities, the right ventricular cavity was found to be larger than the left ventricular cavity. Intraventricular septum thickness was best measured by the two-dimensional technique, whereas posterior wall dimensions were difficult to assess by this technique. This is the first study that attempts to validate echocardiography as it pertains to the fetus. It shows that a good correlation between invasive and noninvasive methodology can be obtained. Encouraging findings may lead other investigators to use such techniques in the human fetus to quantify ventricular size and output under normal and abnormal conditions.     

Sources and disposition of fetal glucose: studies in the fetal lamb

Three aspects of fetal glucose metabolism are studied in the pregnant sheep: source of fetal glucose, rates and routes of fetal glucose disposal, and how these processes are regulated in the fetus. However, extrapolation of findings in the fetal sheep to the condition existing in the human must be made with great caution. This is mainly due to the fact that sheep are ruminants that obtain much of their energy from volatile fatty acids and fetal sheep have low glucose and high fructose concentrations.     

Physiologic baroreceptor activity in the fetal lamb.

OBJECTIVE: We investigated the influences of gestational age and behavioral state on fetal baroreceptor activity. STUDY DESIGN: Two protocols were used to analyze low-voltage fast activity and high-voltage slow activity in eight chronically instrumented ovine fetuses at days 127 to 142. In protocol 1 correction of fetal heart rate to mean arterial blood pressure was assessed under physiologic conditions, and the linear regression slope of fetal heart rate on mean arterial pressure was calculated. In protocol 2 an injection of 7.5 micrograms of norepinephrine was given during consecutive behavioral cycles. The ratio of fetal heart rate decrease to mean arterial pressure increase was determined as representing baroreceptor activity. RESULTS: Under protocol 1 the slope was steeper in high-voltage slow activity (2.33 +/- 1.03) than in low-voltage fast activity (0.89 +/- 0.37) (n = 21, p less than 0.001), indicating that fetal heart rate changes are sensitive to blood pressure changes during high-voltage slow activity. Under protocol 2 inverse relationships were found between the ratio of fetal heart rate decrease to mean arterial pressure increase and gestational age in low-voltage fast activity and high-voltage slow activity. CONCLUSION: Fetal baroreceptor activity increased during high-voltage slow activity and decreased with gestational age.     

Umbilical artery pulsatility index and placental vascular resistance during acute hypoxemia in fetal lambs

The effect of hypoxemia on the pulsatility index (PI) of the umbilical artery flow velocity waveform and placental vascular resistance was studied. Fetal hypoxemia was induced by maternal breathing of a low-oxygen gas mixture. Umbilical venous blood flow was measured with an electromagnetic flowmeter. Placental vascular resistance (PVR) was defined as the ratio perfusion pressure (mean arterial pressure minus umbilical venous pressure) and umbilical blood flow. Umbilical artery velocity waveforms were obtained by a 5-MHz pulsed Doppler device around one umbilical artery in 4 lambs and by a transcutaneous 4-MHz continuous wave Doppler transducer in 3 lambs. Fetal arterial oxygen content was lowered from 2.28 +/- 0.18 to 0.93 +/- 0.15 mM (p less than 0.05), while pCO2 and pH remained unchanged. Control values of the hemodynamic variables were compared with values during deepest hypoxemia. Fetal heart rate, mean arterial and umbilical venous pressure, PVR and the umbilical artery PI did not significantly change, whereas umbilical blood flow increased from 436 +/- 64.7 to 491 +/- 65.9 ml/min (p less than 0.05) during deepest hypoxemia. Individual regression analysis, however, showed a significant inverse correlation of umbilical venous pressure whereas PVR had a positive correlation with actual oxygen content. It is concluded that acute fetal hypoxemia slightly decreases PVR, but does not affect the umbilical artery PI in sheep. Decreasing fetal oxygenation is associated with an increase in pressure in the umbilical vein.     

Relationship between blood gas values and hormonal response to acute hypoxemia in fetal sheep

We examined the relationship of fetal plasma immunoreactive adrenocorticotropic hormone (IR-ACTH), arginine vasopressin (AVP) and cortisol to fetal arterial pO2 (PaO2), oxygen saturation (SaO2), change (delta) in PaO2 (delta PaO2) or SaO2 (delta SaO2) in fetal sheep at 125-139 days of gestational age. Fetal sheep were exposed to two levels of hypoxemia induced by altering the inspired percent oxygen to the mother for 1 h without fetal acidemia. Fetal carotid arterial IR-ACTH, AVP and cortisol concentrations during hypoxemia were correlated with fetal PaO2, SaO2, delta PaO2 and delta SaO2 by applying simple regression analysis. For IR-ACTH, a strong correlation was obtained with SaO2, delta SaO2 and delta PaO2 after log transformation to IR-ACTH. For AVP, strong correlation coefficients were obtained with delta SaO2, SaO2, delta PaO2 and PaO2 after reciprocal transformation to AVP, but for cortisol a strong correlation was obtained only with delta PaO2 after log transformation to cortisol. We conclude that during fetal hypoxemia without acidemia, there is a strong correlation between IR-ACTH and SaO2, delta SaO2, delta PaO2 but not PaO2, between AVP and delta SaO2, SaO2, delta PaO2, or PaO2, and between cortisol and delta PaO2.     

Ultrasound evaluation of tachycardia-induced hydrops in the fetal lamb

Nonimmune hydrops was induced in six fetal lambs at 120 days' gestation by right atrial pacing at 300 beats/min. Biochemical, hemodynamic, and Doppler parameters were observed and monitored by real-time ultrasound during the creation (18 to 42 hours) and resolution of hydrops. The significant changes seen were hypoproteinemia (34.3 to 29.0 mg/100 ml), venous hypertension (24.6 to 31.3 mm Hg), decreases in thoracic aorta flow, and decreases in systolic/diastolic ratios in the aorta and umbilical artery. These changes were reversed with the discontinuation of pacing and the resolution of fetal hydrops.     

The effect of electrical stimulation on behavioral states in the fetal lamb

Brachial nerve stimulation at two frequencies (0.01 and 0.05 pulse per second, pps) with the fetus in either low-voltage electrocortical activity with eye movements or high-voltage electrocortical activity without eye movements was studied in five chronically catheterized fetal lambs at 130 to 140 days' gestation. During low-voltage electrocortical activity with eye movements, there were slight alterations in electrocortical state and electromyographic activity at 0.01 pps. At 0.05 pps these electromyographic changes were enhanced above those expected as a result of state changes but returned to control values during the recovery period. During high-voltage electrocortical activity without eye movements, the state changes were much more dramatic, while electromyographic activity increased significantly only at 0.05 pps. It was of note that in both high- and low-voltage electrocortical activity the fetal heart rate changed only during the recovery period at 0.05 pps. It is concluded that fetal behavioral state influences the interpretation of biophysical measurements in the fetus and that the effect of stimulation is more pronounced if applied when the fetus is in high-voltage electrocortical activity without eye movements.     

Amniotic gas values and acid-base status during acute maternal hyperoxemia and hypoxemia in the early fetal sheep

OBJECTIVE: This study was undertaken to determine amniotic fluid gas values and acid-base balance during maternal hyperoxemia and hypoxemia in early pregnancy. STUDY DESIGN: Anesthetized sheep (n = 12) in early and mid pregnancy (0.3 to 0.5 gestation) were subjected to hyperoxemia followed by hypoxemia. Amniotic fluid PO (2), pH, PCO (2), bicarbonate concentration, and base excess were monitored continuously with a multiparameter Paratrend (Diametrics Medical Inc, St Paul, Minn) sensor and compared between 0.3 and 0.5 gestation. RESULTS: During maternal normoxemia all parameters were constant. At all gestational ages maternal hyperoxemia caused no changes apart from a rapid increase in amniotic fluid PO (2) (P <.001). Maternal hypoxemia led to a reduced amniotic fluid PO (2) (P <.001), whereas the PCO (2) and the bicarbonate concentration increased (P <.001). Changes in amniotic fluid gas values and acid-base balance were more pronounced at 0.3 gestation than at 0.5 gestation. Amniotic fluid Po(2) responded earlier to maternal hyperoxemia than to hypoxemia (9. 5 vs 14.3 minutes; P <.001). During hypoxemia maternal PaCO (2) changed faster than did amniotic fluid PCO (2) (P <.001). CONCLUSIONS: Acute maternal hypoxemia during early pregnancy was quickly reflected in amniotic fluid gas values and acid-base balance, whereas hyperoxygenation induced the quickest changes in amniotic fluid PO (2). The metabolic effects of maternal hyperoxemia and hypoxemia were also more pronounced during early pregnancy than at mid pregnancy.     

Experimentally produced sinusoidal fetal heart rate pattern in the chronically instrumented fetal lamb

A sinusoidal fetal heart rate and elevated fetal arginine vasopressin levels were found in two bled fetal lambs. Arginine vasopressin was continuously infused intravenously in chronically instrumented fetal lambs at rates ranging between 1.0 and 90 mlU/kg/min. No sinusoidal heart rate occurred in 21 experiments on six control animals with intact vagal nerves. Atropine at doses less than 0.4 mg had no effect (six experiments on three animals). Higher doses of atropine (greater than 1.2 mg) resulted in sinusoidal heart rate (24 experiments on six animals). Twenty-three infusions were performed in five bilaterally vagotomized fetuses, and the sinusoidal heart rate was successfully induced in 17 experiments. The arginine vasopressin infusion rate was less than 1.5 mlU/kg/min on the six occasions in which the sinusoidal heart rate did not appear. All sinusoidal heart rate patterns were associated with fluctuation of fetal arterial pressure at the same frequency. There was a linear correlation between the amplitude of sinusoidal heart rate and magnitude of arterial blood pressure fluctuation. The amplitude of sinusoidal heart rate increased with the concentration of arginine vasopressin infused. Modification of sinusoidal heart rate pattern was attempted with use of sympathetic agonists and antagonists. The possible etiology and mechanisms of sinusoidal heart rate pattern are discussed.