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We showed that antenatal corticosteroids reduced blood-brain barrier permeability in fetuses at 60 and 80%, but not 90% of gestation, and decreased brain water content in fetuses. Our objective was to examine the effects of postnatal corticosteroids on regional blood-brain barrier permeability and brain water content in newborn lambs. Three dexamethasone treatment groups were studied in 3- to 5-day-old lambs. A 0.01 mg/kg dose was selected to estimate the amount of dexamethasone that might have reached fetuses via antenatal treatment of ewes in our previous studies. The other doses (0.25 and 0.5 mg/kg) were chosen to approximate those used clinically to treat infants with bronchopulmonary dysplasia. Lambs were randomly assigned to receive four intramuscular injections of dexamethasone or placebo given 12 h apart on days 3 and 4 of age. Blood-brain barrier function was measured with the blood-to-brain transfer constant (K(i)) to alpha-aminoisobutyric acid, brain plasma volume was measured with polyethylene glycol for the calculation of K(i,) and brain water was measured by wet-to-dry tissue weights. Postnatal treatment with corticosteroids did not reduce barrier permeability in newborn lambs. Brain blood volume was higher in the 0.25 and 0.5 mg/kg dose dexamethasone groups than in the placebo group. Brain water content did not differ among the groups. We conclude that postnatal treatment with corticosteroids did not reduce regional blood-brain barrier permeability or brain water content but increased the brain plasma volume in newborn lambs. These findings are consistent with our previous work indicating that barrier permeability is responsive to corticosteroids at 60 and 80% of gestation and brain water regulation at 60% of gestation, but not in near-term fetuses or newborn lambs.

The objective of the present study was to examine the effect of antenatal or postnatal treatment with corticosteroids on the NMDA receptor, one of the mediators of both normal brain development and hypoxic-ischemic injury, by determining the characteristics of the receptor MK-801 binding site in untreated and corticosteroid-treated fetal and newborn lambs. (3)H-MK-801 binding was performed in cerebral cortical cell membranes from fetal sheep at 88, 120, and 136 d gestation (term = 150 d), and from 5-d-old lambs and adult ewes. Animals were randomized to receive dexamethasone [fetuses: 6 mg, i.m. every 12 hr for four doses to mother; lambs: 0.01 mg/kg (low dose) or 0.25 mg/kg (high dose) every 12 hr for four doses] or placebo. During development, B(max) (apparent number of receptors) increased, reaching a maximum in 5-d-old lambs (p < 0.05) and decreasing in the adult brain. K(d) (dissociation constant) did not change, suggesting that receptor affinity was not altered during maturation. Dexamethasone treatment had no effect on MK-801 binding in the fetus or adult, but in lambs was associated with a significant decrease in B(max) from 2.17 +/- 0.18 pmol/mg protein in placebo-treated animals to 1.65 +/- 0.8 and 1.62 +/- 0.07 pmol/mg protein in low-dose and high-dose animals, respectively. Affinity for (3)H-MK-801 decreased 20% after dexamethasone treatment in lambs only (p < 0.05). Thus, dexamethasone treatment appears to modify the NMDA receptor only during a specific period of brain development.

We previously reported decreases in blood-brain barrier permeability in the ovine fetus at 80% of gestation after antenatal corticosteroids and shown that permeability is not reduced in newborn lambs after postnatal corticosteroids. We now test the hypotheses that exogenous antenatal corticosteroids decrease blood-brain barrier permeability at 60% but not 90% of gestation in ovine fetuses and that endogenous increases in plasma cortisol concentrations are associated with ontogenic decreases in barrier permeability during gestation. Chronically instrumented ovine fetuses were studied 12 h after the last of four 6-mg dexamethasone or placebo injections were given 12 h apart over 48 h to ewes. Fetuses at 80% of gestation from placebo-treated ewes studied under the same protocol were also included. Blood-brain barrier function was quantified with the blood-to-brain transfer constant (K(i)) to alpha-aminoisobutyric acid. K(i) values were lower in cerebral cortex, caudate nucleus, hippocampus, superior colliculus, thalamus, medulla, and cervical spinal cord in fetuses of dexamethasone- than placebo-treated ewes at 60% but not 90% of gestation. Regional brain K(i) values demonstrated inverse correlations with increases in gestation and plasma cortisol concentrations in most brain regions. We conclude that maternal treatment with exogenous corticosteroids was associated with decreases in blood-brain barrier permeability at 60% but not 90% of gestation and that increases in gestation and endogenous cortisol concentrations were associated with ontogenic decreases in barrier permeability during fetal development.

We studied the effects of hypoxia on cerebral cortical and intestinal perfusion and metabolism in normocythemic hyperviscous newborn pigs. Seven pigs were made hyperviscous by an injection of cryoprecipitate, increasing viscosity from 5.8 +/- 0.9 to 9.0 +/- 1. 2 (SD) cycles/s. Six normoviscous pigs received 0.9% NaCl. Reducing the inspired O(2) decreased the arterial O(2) content (Ca(O(2))) from 9.5 +/- 1.6 to 3.6 +/- 1.3 ml O(2)/100 ml. Increases in brain and decreases in gastrointestinal blood flow at the lower Ca(O(2)) values were similar between the groups. During hypoxia, blood flow to stomach, distal intestinal mucosa, and large intestines was lower (-50, -23, and -28%, respectively) in the hyperviscous than normoviscous group. At the lower Ca(O(2)) values, cerebral cortical vascular resistance decreased in both groups and intestinal vascular resistance increased (+257%) in the hyperviscous but not in the normoviscous group. During hypoxia, systemic oxygen delivery decreased, extraction increased, and uptake did not change; cerebral cortical O(2) delivery, extraction, and uptake did not change; and intestinal O(2) delivery decreased, extraction increased, and uptake did not change in both groups. Our study demonstrated that 1) during hypoxia, increases in systemic O(2) extraction compensated for decreases in delivery and systemic uptake did not change; vasodilation sustained cerebral cortical O(2) delivery and preserved metabolism; increases in intestinal oxygen extraction offset decreases in delivery and uptake was preserved; and 2) nonpolycythemic hyperviscosity did not have a major influence on cardiovascular or metabolic responses to hypoxia, except for modest effects on intestinal resistance and perfusion to certain gastrointestinal regions. We conclude that, under normocythemic conditions, a moderate increase in viscosity does not have a major impact on hemodynamic or metabolic adjustments to hypoxia in newborn pigs.

We have been studying the ontogeny of the blood-brain barrier function in ovine fetuses and lambs. During these studies, we have found that the duration of ventilation also influences blood-brain barrier permeability in premature lambs. Chronically instrumented hysterotomy-delivered surfactant-treated premature lambs were studied at 90% or 137 days of gestation (n = 9). Blood-brain barrier function was quantified with the blood-to-brain transfer constant K(i) to alpha-aminoisobutyric acid. Linear regression analysis was used to compare the K(i) values in the brain regions, as the dependent variable, to the duration of ventilation, as the independent variable. There were direct correlations (P < 0.05) between the K(i) values and the duration of ventilation [306 min (mean), 162-474 min (range)] in the cerebral cortex, cerebellum, medulla, caudate nucleus, hippocampus, superior colliculus, inferior colliculus, thalamus, pons, cervical spinal cord, and choroid plexus, but not in the pituitary gland. Ventilatory pressures and rates were established before the onset of the permeability studies. Calculated mean airway pressures [14 cmH(2)O (mean), 7-20 cmH(2)O (range)] from similarly studied premature lambs did not correlate with the duration of positive-pressure ventilation. We conclude that increases in the duration of positive-pressure ventilation predispose premature lambs to increases in regional blood-brain barrier permeability. These alterations in barrier function occur over relatively short time intervals (minutes to hours). In our study, these changes in permeability are most likely not attributable to changes in mean airway pressure.

The purpose of this study was to characterize postnatal changes in regional Doppler blood flow velocity (BFV) and cardiac function of very-low-birthweight infants and to examine factors that might influence these hemodynamic changes. Mean and end-diastolic BFV of the middle cerebral and superior mesenteric arteries, cardiac output, stroke volume, and fractional shortening were measured in 20 infants birthweight 1,002 +/- 173 g, gestational age 28 +/- 2 wk) at 6, 30, and 54 h after birth and before and after feedings on days 7 and 14. Postnatal increases in cerebral BFV, mesenteric BFV, and cardiac output were observed that were not associated with changes in blood pressure, hematocrit, pH, arterial PCO(2), or oxygen saturation. The postnatal pattern of relative vascular resistance (RVR) differed between the cerebral and mesenteric vasculatures. RVR decreased in the middle cerebral but not the superior mesenteric artery. Physiological patency of the ductus arteriosus did not alter postnatal hemodynamic changes. In response to feeding, mesenteric BFV and stroke volume increased, and mesenteric RVR and heart rate decreased. Postprandial responses were not affected by postnatal age or the age at which feeding was initiated. However, the initiation of enteral nutrition before 3 days of life was associated with higher preprandial mesenteric BFV and lower mesenteric RVR than was later initiation of feeding. We conclude that in very-low-birthweight infants over the first week of life 1) systemic, cerebral, and mesenteric hemodynamics exhibit region-specific changes; 2) asymptomatic ductus arteriosus patency and early feedings do not significantly influence these postnatal hemodynamic changes; and 3) cardiac function adapts to increase local mesenteric BFV in response to feedings.

We tested the hypotheses that, in hypoxic young pigs, reductions in cardiac output restrict systemic oxygen transport to a greater extent than does hypoxia alone and that compensatory responses to this restriction are more effective in higher than in lower priority vasculatures. To study this, 10- to 14-day-old instrumented awake hypoxic (arterial oxygen tension = 39 Torr) pigs were exposed to reduced venous return by inflation of a right atrial balloon-tipped catheter. Blood flow was measured with radionuclide-labeled microspheres, and oxygen metabolism was determined with arterial and venous oxygen contents from appropriate vessels. Hypoxia resulted in a reduction in oxygen tension; increases in cardiac output and perfusion to brain (72% over baseline), heart, adrenal glands, and liver without reductions to other organs except for the spleen; reductions in systemic and intestinal oxygen delivery; and increases in systemic and intestinal oxygen extraction without changes in systemic, cerebral, or intestinal oxygen uptake. During hypoxia, decreasing venous return was associated with increases in arterial lactic acid concentration and central venous pressure; attenuation of the hypoxia-related increase in cardiac output; sustained increases in brain (72% over baseline) and heart perfusion; reductions in lung (bronchial artery), pancreatic, renal, splenic, and intestinal (-50% below baseline) perfusion; decreases in systemic and gastrointestinal oxygen delivery; sustained increases in systemic and intestinal oxygen extraction; and decreases in intestinal oxygen uptake, without changes in cerebral oxygen metabolism. We conclude that when venous return to the heart is reduced in hypoxic young pigs, the hypoxia-related increase in cardiac output was attenuated and the relative reduction in cardiac output was associated with preserved cerebral oxygen uptake and compromised intestinal oxygen uptake. Regional responses to hypoxia combined with relative reductions in cardiac output differ from that of hypoxia alone, with the greatest effects on lower priority organs such as the gastrointestinal tract.

Although the pathogenesis of necrotizing enterocolitis remains uncertain, ischemia appears to be an important contributing factor to the development of this disorder. Reperfusion plays a major role in ischemia-related injury, and oxygen free radicals produced during reperfusion most likely contribute to the injury. These oxidants can be generated during prostanoid metabolism, which increases during reperfusion of ischemic gut in adult subjects. The present study was designed to: 1) examine the effects of superior mesenteric artery occlusion, e.g. ischemia and reperfusion in vivo on the development of histopathologic intestinal injury; 2) determine whether products of arachidonic acid metabolism, e.g. prostanoids are increased during reperfusion of ischemic gut; and 3) determine whether oxygen free radical scavengers attenuate the injury in newborn pigs. Chronically catheterized placebo-pretreated newborn pigs exposed to ischemia-reperfusion, placebo-pretreated nonischemic control pigs, and polyethylene glycol-superoxide dismutase (SOD) plus polyethylene glycol-catalase (CAT)-pretreated, ischemic pigs were studied by examining changes in intestinal circulation, oxygenation, prostanoids, and tissue injury. In the placebo-pretreated pigs, intestinal blood flow decreased to very low levels during superior mesenteric artery occlusion. During reperfusion, blood flow increased, but remained below baseline. After ischemia, oxygen uptake returned to values that were similar to baseline. Intestinal efflux of the vasodilator 6-keto-prostaglandin F1alpha was evident (p < 0.05 versus no or zero efflux) during early reperfusion. Histopathologic scoring of terminal ileal samples showed significant mucosal necrosis, surface epithelial disruption, lamina propria congestion and hemorrhage, submucosal hemorrhage, edema, and increases in cells compared with the placebo-pretreated nonischemic pigs. In the SOD plus CAT-pretreated ischemic pigs, changes in intestinal blood flow, oxygen uptake, 6-keto-prostaglandin F1alpha efflux, and the pattern of the ileal tissue injury did not differ significantly from the placebo-pretreated ischemic pigs. In summary, superior mesenteric artery occlusion for 1 h and reperfusion for 2 h resulted in severe intestinal ischemia, early postocclusive limited increases in intestinal perfusion and oxygen uptake, efflux of vasodilating prostanoids during early reperfusion, and signs of ischemic tissue injury in the placebo- and SOD plus CAT-pretreated pigs. This study demonstrates that, after superior mesenteric artery occlusion and reperfusion, severe intestinal tissue injury is detected in vivo, prostanoid efflux increases, and SOD plus CAT given just before occlusion does not attenuate the extent of injury in newborn pigs.

Fetuses of women whose diabetes is poorly controlled often exhibit hypoxemia and elevated catecholamine concentrations at birth. In the ovine fetus, experimental hyperinsulinemia results in hypoxemia, elevated catecholamine concentrations, and hemodynamic changes. Limited oxygen availability occurring during pregnancy-related complications and/or delivery may present an additional risk to the hyperinsulinemic fetus. In this study, we tested the hypothesis that hypoxia induced via acutely limiting oxygen availability compromises the hemodynamically and metabolically stressed but compensated hyperinsulinemic ovine fetus. Fetuses receiving insulin (n = 8) or placebo (n = 5) for 48 h were exposed to maternally induced hypoxia. Hypoxic hypoxia was associated with a surge in catecholamines in the hyperinsulinemic fetuses. During hypoxia, this group exhibited insulin-related sustained increases in the combined ventricular output and fetal body blood flow, accentuation of the prior insulin-related increase in blood flow to the heart, decreased systemic oxygen delivery, accentuation of the insulin-related increased oxygen extraction, reductions in the insulin-related increased systemic oxygen uptake, sustained increases in regional oxygen delivery to the heart and adrenal glands, reductions in the insulin-related increased delivery to the carcass, and decreased oxygen delivery to the kidneys and gastrointestinal tract. We conclude that, in the hyperinsulinemic ovine fetus, hypoxic hypoxia attenuates the hyperinsulinemia-mediated increased systemic oxygen uptake. Regional oxygen transport is preserved to vital regions (brain, heart, and adrenal glands) by increased perfusion and compromised to certain other regions (kidneys and gastrointestinal tract), because the increases in perfusion are insufficient to offset the limited oxygen availability.

Newborn infants of diabetic mothers have serum biochemical signs of iron deficiency in cord blood directly related to elevations of cord erythropoietin and Hb concentrations. In sheep, chronic fetal hyperinsulinemia results in fetal hypoxemia, expansion of the red cell mass, and decreased iron concentrations, most likely due to increased iron utilization for Hb synthesis. To determine whether fetal insulin exposure also results in reduced tissue iron concentrations, we measured liver, skeletal muscle, small intestine, heart, and brain iron concentrations in newborn rat pups after s.c. fetal injection of insulin or diluent alone on d 19 of gestation. The fetuses of 11 pregnant rats were exteriorized, injected with 2 U neutral protamine Hagedorn insulin or diluent, replaced in utero, and delivered on d 22. To determine dose dependency, the fetuses of six pregnant rats were injected with 3 U of longer-acting protamine zinc insulin and delivered on d 21. At delivery, the insulin-treated groups had higher birth weights than the placebo-treated group, although plasma insulin concentrations were not different. The 2 U neutral protamine Hagedorn insulin-treated fetuses had significantly lower mean +/- SEM liver iron concentrations than the control fetuses (910 +/- 34 versus 1014 +/- 43 micrograms/g dry tissue weight; p less than 0.05), but had similar skeletal muscle iron concentrations. The 3 U protamine zinc insulin-treated fetuses had significantly lower liver and skeletal muscle iron concentrations compared to control and to 2 U neutral protamine Hagedorn insulin-treated fetuses (p less than 0.05). No differences in small intestine, heart, or brain iron concentrations were seen among groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Autoregulation of cerebral blood flow is impaired in the postictal state. This loss of autoregulation may in part be mediated by a rise in perivascular hydrogen ion and carbon dioxide concentration. We hypothesized that hypocarbia with a concomitant reduction in perivascular hydrogen ion and carbon dioxide concentration would restore autoregulation during the postictal state. We studied autoregulation of cerebral blood flow in 13 ventilated, awake 3- to 4-d-old piglets during the postictal state under normocarbic and hypocarbic conditions. During the postictal state, cerebral blood flow was pressure-passive in normocarbic piglets, whereas the relationship between cerebral blood flow and cerebral perfusion pressure was described by a polynomial curve in hypocarbic piglets. Because hypocarbia restores cerebral blood flow autoregulation in postictal newborn piglets, we speculate that the perivascular hydrogen ion and carbon dioxide concentration contribute significantly to the state of cerebral autoregulation in the postictal subject.

The effects of acute maternal hemorrhage on uterine blood flow and fetal circulation were investigated in pregnant sheep. Nine chronically instrumented pregnant sheep (114-128 d gestation), phlebotomized from the iliac artery at the point of origin of the uterine artery, were studied at baseline, after acute hemorrhage, and immediately and two h after replacement of the blood. Maternal hemorrhage caused a reduction in uterine blood flow as well as both fetal hypoxemia and acidemia. Changes in fetal organ blood flow measured by radionuclide-labeled microspheres showed that blood flow to the brain, heart, and adrenal glands increased (p less than 0.05), whereas blood flow to the other major organs did not change significantly. Rapid replacement of blood restored all parameters to baseline values. We conclude that acute maternal hemorrhage causes a reduction in uterine blood flow, fetal hypoxemia, and acidemia with a secondary increase in blood flow to the high priority organs. Rapid replacement of blood reverses these effects.

Posthepatic insulin availability has been evaluated by steady-state insulin turnover studies with 131I-insulin. Spontaneously delivered term (age 3.3 +/- 0.8 days) (mean +/- S.E.) and prematurely delivered lambs (betamathasone treated at 132 days) (age, 1.1 +/- 0.2 days) were compared with 4- to 5-month-old sheep. After a 7-hr fast, animals received 0.45% saline or 5.7 mg/kg/min glucose (0.06 ml/kg/min) for 6-hr followed by the tracer insulin infusion for 110 min. Plasma glucose, insulin, and immunoprecipitable insulin were measured sequentially during the steady state. Endogenous posthepatic insulin secretion and metabolic clearance rates were derived. Neither endogenous posthepatic insulin secretion rates nor metabolic clearance rates were different among the three groups of animals when either 0.45% saline or 5.7 mg/kg/min exogenous glucose infusions were compared. Secretory response of the pancreatic beta cell to continuous glucose infusion seems similar for the term and preterm lamb when compared to adult sheep.

We tested the hypothesis that isovolemic anemia blunts the thermogenic response to environmental cold stress in 3 to 4-day-old newborn piglets. Eight animals were studied in both thermoneutral (31.6-32.8 degrees C) and cold (19.6-20.2 degrees C) environments, before and after an isovolemic, partial volume exchange transfusion which reduced the hematocrit from 26 to 15%. In the nonanemic phase of study, deep rectal temperatures declined but had plateaued by 30 minutes after onset of cold stress and remained within normal limits for newborn piglets. In the anemic phase of study, deep rectal temperature declined continuously throughout cold stress with true body core hypothermia (less than 38 degrees C) observed at all measurement points beyond 15 min of cold stress. Baseline oxygen consumption did not differ between the two study phases (17.6 +/- 1.8 versus 16.7 +/- 2.1 ml/kg-1/min-1, mean +/- SEM). However, during environmental cold stress, oxygen consumption increased by 64% over baseline in the nonanemic phase of study (p less than 0.05) whereas 21% increase over baseline was observed in the anemic phase (p NS). We conclude that isovolemic anemia limited oxygen consumption and heat production during environmental cold stress, resulting in body core hypothermia.

The effects of severe hypoxemia on gastrointestinal (GI) blood flow and gastric emptying were studied in nine 2- to 4-day-old piglets which were mechanically ventilated while receiving nitrous oxide anesthesia. Each animal was studied during a control period of oxygenation (PaO2 91 +/- 8 torr), 35 min of hypoxemia (PaO2 29 +/- 1 torr), and a recovery period (PaO2 90 +/- 5 torr) (mean +/- SEM). During each study period, the animal received a 10% dextrose test meal with phenol red marker (22 ml/kg), gastric residual volumes were determined at 10-min intervals over 30-min study periods using a dye dilution double sampling technique, and GI blood flow (radionuclide-labeled microspheres), O2 delivery, O2 extraction, and O2 consumption were measured at the end of the 30-min period. Hypoxemia resulted in decreased blood flow to the following GI organs: stomach, jejunal and ileal mucosa-submucosa, and colon decreased 62, 31, and 35%, respectively (p less than 0.05). Jejunal and ileal muscularis blood flow remained unchanged. Oxygen delivery and consumption by GI tract decreased 79 and 58%, respectively; whereas oxygen extraction of GI tract increased 115%. Values returned toward baseline levels during the recovery period. The hypoxemic gastric emptying pattern showed significantly greater gastric residuals at 20 min compared to the 10-min value (p less than 0.05). This pattern was different than that observed during control and recovery periods. We conclude that severe hypoxemia results in decreased GI blood flow, tissue oxygenation, and an altered gastric emptying pattern. These observations may have clinical significance for feeding infants following an hypoxemic episode.

Despite methodologic problems, endogenous creatinine clearance is commonly used as an estimation of glomerular filtration rate (GFR). Inulin clearance was compared to endogenous creatinine clearance in a group of low birthweight infants to establish the validity of the latter. Thirty-three low birthweight infants (birthweight mean = 1600 g, gestational age mean = 33 wk) were studied between 10 hr and 10 days of age to simultaneously measure GFR by inulin and endogenous creatinine clearances. Inulin and creatinine clearances correlated directly (r = 0.738, P greater than 0.001). The slope of the regression line suggested an overestimation of GFR (inulin clearance) by creatinine clearance at the low GFR range and an underestimation at the high GFR range. The data were divided into two groups by the median inulin clearance (12.5 ml/min/1.73m2). The ratio of creatinine to inulin clearance was significantly higher in the low GFR group (1.28 +/- 0.16 vs. 0.89 +/- 0.04 SEM, n = 19, P less than 0.05). There was no difference between the two groups in plasma creatinine, birthweight, gestational age, incidence of respiratory distress, or oxygen requirements at the time of the studies. Endogenous creatinine clearance represents a good estimation of GFR (inulin clearance) in low birthweight infants. However, at the low GFR range, it represents an overestimation and at the high GFR range, an underestimation.

The effects of feeding on gastrointestinal (GI) perfusion and oxygen transport in hypoxemic neonates is unknown. We evaluated these effects in unanesthetized, spontaneously breathing newborn piglets by comparing three experimental groups: nine hypoxemic piglets (mean PaO2 26 torr) which were fed with formula, six hypoxemic piglets (mean PaO2 27 torr) which were not fed, and four normoxemic piglets (mean PaO2 79 torr) which were fed and served as controls. The control-fed group exhibited an increase in stomach and small intestinal mucosal-submucosal blood flow within 30 min following feeding which was significantly greater than that observed in the hypoxemic fed piglets. GI O2 delivery and O2 uptake rose significantly (p less than 0.05) following a meal secondary to increases in total GI blood flow. Oxygen extraction was unchanged postprandially in the control group. In the hypoxemic nonfed piglets, total and regional GI blood flow was unaltered during hypoxemia. Reductions in arterial O2 content led to significant decreases in GI O2 delivery. Gastrointestinal oxygen uptake remained stable with a compensatory increase in GI O2 extraction. In the hypoxemic-fed piglets, hypoxia significantly decreased stomach blood flow and led to unchanged blood flow in the remainder of the GI tract. Significant reductions in arterial O2 content and GI O2 delivery were observed, accompanied by significant increases in O2 extraction. Hypoxemic fed animals did not exhibit the expected increase in O2 uptake to meet postprandial metabolic demands. When the hypoxemic insult was terminated, fed piglets demonstrated significant total and regional GI hyperemia leading to increased GI O2 uptake when compared with hypoxemic nonfed piglets.(ABSTRACT TRUNCATED AT 250 WORDS)

Since kernicteric lesions are usually found in the subcortical regions of the brain and these areas also receive the highest blood flow during asphyxia and hypercapnia, we hypothesized that increases in brain bilirubin deposition may be related to increases in brain blood flow. Fourteen piglets underwent a 3-h infusion of bilirubin to maintain total serum bilirubin at approximately 8 mg/dl, during which time blood gases, hemodynamic variables, and brain blood flow were determined. After sacrificing the animals, regional brain bilirubin content was determined. Ten piglets underwent the same protocol; in addition, hypercapnia was induced during the last hour of study (PaCO2 approximately 70 mm Hg). The regional brain blood flow and bilirubin deposition were significantly increased over control values (p less than 0.05) following hypercapnia in the subcortical region and significantly so in the midbrain and cerebellum. In separate groups of control (n = 6) and hypercapnia (n = 6) piglets, 125I-labeled albumin was infused and demonstrated that hypercapnia was not associated with increased regional brain albumin content. We conclude that hypercapnia-induced augmentation in regional brain blood flow is associated with increased deposition brain blood flow is associated with increased deposition of unbound bilirubin. Although the causal relationship between these two observations has not been firmly established, the findings deserve future investigation to clarify the role of brain blood flow, brain bilirubin deposition, and the production of kernicterus in high risk infants.

Permeability of the blood brain barrier (BBB) for bilirubin and 125I-albumin was studied in 2-d- and 2-wk-old piglets. 125I-albumin was given by bolus at the beginning of each study. Hyperbilirubinemia was produced by an initial bolus infusion of bilirubin and sustained at a plasma bilirubin:albumin molar ratio of approximately 1.0 by continuous infusion of bilirubin for 3 h. During the study period, arterial pH and blood gas tensions, serum osmolarity, and mean arterial blood pressure were within the physiologic range for age in both groups. Serum albumin and serum total and unbound bilirubin concentrations were higher in the 2-wk-old piglets. Brain bilirubin concentrations and permeability (P.S product) of the BBB for bilirubin were higher in the 2-d-old than in the 2-wk-old piglets, but the values of P.S for albumin were not different between the two groups. In 2-d-old piglets, regional brain bilirubin concentrations and permeability of the BBB were higher in subcortical regions (cerebellum and brainstem) than in the cerebral cortex. Regional brain albumin concentrations and BBB permeability to albumin in 2-d-old piglets were higher only in the cerebellum. In all regions, the bilirubin:albumin molar ratio was higher in the brain tissues than in the blood. In 2-wk-old piglets, the brain concentrations and P.S products for bilirubin were lower and the regional differences were less marked than for 2-d-old animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Regional and total gastrointestinal (GI) blood flow, O2 delivery, O2 extraction, and O2 consumption were measured before and after acute blood volume expansion in 2-day-old piglets. Blood flow was measured with radionuclide-labeled microspheres. Sixty minutes after a rapid transfusion of age- and hematocrit-matched piglet donor blood, 51Cr-measured blood volume increased 19% while an increase in hematocrit suggested plasma transudation to the extravascular space had occurred in response to blood volume loading. Following transfusion, total GI blood flow and O2 delivery did not change, although O2 extraction decreased by 31 +/- 4%. O2 consumption by the GI tract decreased from 2.0 +/- 0.19 ml O2 X min-1 X 100 g-1 to 1.46 +/- 0.24 ml O2 X min-1 X 100 g-1 1 h after transfusion. Feeding was then accomplished via orogastric tube to determine if animals stressed by blood volume loading would increase GI O2 consumption in response to feeding. The postprandial increase in GI O2 consumption was similar to that previously reported in newborn piglets. Therefore, in the fasting state, acute blood volume loading disrupts GI O2 transport at the capillary level and decreases GI O2 consumption. However, animals subjected to blood volume loading appear capable of increasing GI O2 consumption after feeding.

Acute hyperosmolality-induced blood brain barrier breakdown has been demonstrated to increase the permeability of sucrose, which is similar in molecular weight to bilirubin, independently of changes in regional brain blood flow. We studied three groups of piglets given continuous bilirubin infusions to maintain serum bilirubin concentrations at approximately 8 mg/dl. Normal serum osmolality was maintained throughout the study in control animals. Two experimental groups were made hyperosmolar (330 and 375 mosmol/liter) with bolus urea infusions during the last hour of the study. Regional brain bilirubin concentrations were elevated in the 375 mosmol/liter hyperosmolal experimental group, but not in the 330 mosmol/liter group. Regional brain albumin concentrations also were increased over the control group in the 375 mosmol/liter animals. There were no differences in regional brain blood flows to account for the increases in brain bilirubin concentrations. Our results show that brain bilirubin deposition occurs following breakdown of the blood brain barrier by acute, severe hyperosmolality (375 mosmol/liter) and that the deposited bilirubin is derived from both bound and unbound fractions. The bilirubin deposition occurs independently of changes in regional brain blood flow; however, regional differences in the blood brain barrier permeability to albumin also occur.

This study examines the role of impaired gluconeogenesis in the pathogenesis of neonatal hypoglycemia in intrauterine growth retardation (IUGR). IUGR was produced experimentally in eight pregnant rats by ligation of uterine arteries at the 17th day of gestation. Delivery occurred spontaneously at term. Sham operations were performed in five pregnant rats at the same gestational age and the fetuses delivered at term served as controls. The body weights of newborn rats with IUGR were significantly lower than the controls (5.32 +/- 0.12 vs. 6.22 +/- 0.06 g, mean +/- SE, P less than 0.001). The fetal liver weights were also significantly smaller in IUGR than in the control animals (0.224 +/- 0.14 vs. 0.340 +/- 0.12 g, P less than 0.001). The activity of phosphoenolpyruvate carboxykinase (PEPCK) (the rate-limiting enzyme of gluconeogenesis) in liver cytosols was significantly lower in rats with IUGR (0.06 +/- 0.01 vs. 0.11 +/- 0.02 microM phosphoenolpyruvate/g liver/min when compared with controls (P less than 0.05). A direct relationship between this enzyme and the birth weight was observed, suggesting a close relationship between intrauterine nutrition and the status of gluconeogenesis. The blood glucose level was also lower in growth-retarted fetuses (36.6 +/- 4.7 vs. 69.6 +/- 4.3 mg/dl, p less than 0.001) when compared with controls. The data suggest that gluconeogenesis is impaired in IUGR and is partly responsible for the increase in the incidence of neonatal hypoglycemia in this group of subjects.

In adults, exposure to prolonged hypercarbia results in a normalization of the extravascular brain pH associated with a reduction in brain blood flow (BBF). Following prolonged hypercarbia, sudden normalization of the arterial PCO2 also produces a change in the extravascular brain pH to an alkaline state, resulting in a marked decrease in BBF. We examined these physiologic phenomena in newborn subjects by exposing seven awake, spontaneously breathing newborn piglets to 4 h of sustained hypercarbia (PaCO2: 60-70 mm Hg) followed by a 45-min normocarbic period. Total and regional BBF, cardiac output (radionuclide-labeled microsphere method), arterial blood pressure, plasma catecholamine and lactate concentrations, blood gases, oxygen contents, and hematocrits were measured during a baseline period, at 1/2, 2, and 4 h of sustained hypercarbia and 1/4 and 3/4 h following an abrupt onset of normocarbia. The initial 2.5-fold increase in total BBF during hypercarbia persisted for 2 h and at 4 h decreased significantly below the level of the 30-min hypercarbic measurement, although the values still remained 2-fold above the baseline values. Brain tissue pH was reduced at the onset of hypercarbia, remaining unchanged throughout the hypercarbic period. Both total BBF and brain tissue pH returned to baseline values following the return to normocarbia. Changes in regional BBF were similar to that of total BBF with the exception of the boundary zone (periventricular area in the frontoparietal region of the cerebrum, adjacent to the caudate nucleus) and the parietal area (site of the brain tissue pH electrode), where a significant decrease from the peak hyperemia was not observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Bilirubin inhibits in vitro oxidative phosphorylation and glycolysis. This study investigated the in vivo effect of bilirubin on cerebral oxygen, glucose, and lactate uptake in newborn piglets. Seventeen 2- to 4-day-old piglets were divided into three groups and examined as follows: group 1 = control (C); group 2 = control with sulfisoxazole; and group 3 = experimental, given bilirubin with sulfisoxazole. In the experimental group, bilirubin was infused for 4 h. The cerebral bilirubin content in the bilirubin-infused group was 11.0 +/- 1.4 nmol/g of cerebral cortex (mean +/- SEM), consistent with levels found in infants with kernicterus. However, this level of brain bilirubin had no major, acute effects on cerebral uptake of oxygen, glucose, or lactate despite producing lethargy and ataxia which were consistent with bilirubin intoxication. This suggests that mitochondrial changes may not be involved in vivo in acute bilirubin encephalopathy.

The acute effects of normoxemic hypocarbia and hypercarbia were examined in six newborn piglets. Brain blood flow was maintained during hypocarbia until extremely low PaCO2 (less than 15 mm Hg) levels were achieved at which time total brain and cerebral blood flow decreased significantly from baseline values. Blood flow to the thalamus, cerebellum and brain stem was unchanged from baseline conditions during hypocarbia. This suggests that the newborn brain is relatively insensitive to moderate degrees of hypocarbia. Extreme hypocarbia (PaCO2 less than 15 mm Hg) was associated with a significant increase in heart rate, accompanied by a significant decrease in mean arterial blood pressure; however, cardiac output was not significantly different from baseline determinations. Hypercarbia with normoxemia was associated with significant increases in total brain blood flow, with greater blood flow to the brain stem, cerebellum, and thalamus than to the cerebrum. The percentage of cardiac output received by the brain was also significantly increased, although total cardiac output was unchanged. This demonstrates that the newborn cerebral vasculature is sensitive to hypercarbia and that regional differences in sensitivity may account for the greater increments in blood flow to the caudal portions of the brain than that to the cerebrum.

The purpose of this study was to investigate the compensatory change in circulation and oxygenation of the newborn lamb gastrointestinal (GI) tract in response to anemic and hypoxic hypoxemia. Radiolabeled microspheres were used to measure blood flow. We subjected the newborn lamb to a 30-35% reduction in hematocrit 4 d before study and to a 10% oxygen environment for 30 min during the study to induce chronic anemic and acute hypoxic hypoxemia, respectively. The circulatory and oxygenation responses were measured 1 h after a standard milk feeding in all cases. During the experimental periods, no change in total GI blood flow was observed. Because of a failure to augment blood flow during hypoxemia, O2 delivery to the GI tract decreased significantly. Despite this, GI O2 consumption was not compromised because tissue O2 extraction by the GI tract rose significantly. The response of the newborn lamb GI tract to hypoxemia after feeding is augmentation of O2 extraction. The newborn's GI tract did not regulate local GI blood flow.

Newborn infants of poorly controlled insulin-dependent diabetic mothers demonstrate a redistribution of iron from serum and tissue stores into red blood cells. These changes may be due to increases in iron utilization during augmented Hb synthesis, which compensates for chronic intrauterine hypoxemia induced by prolonged fetal hyperinsulinemia. We tested this hypothesis by measuring plasma iron, total iron-binding capacity, percent iron-binding capacity saturation (total iron-binding capacity saturation), Hb concentration, total red cell Hb, and total red cell iron in the arterial blood of 11 chronically instrumented fetal sheep after 7-12 d of infusion with 15 U/day of insulin (n = 5) or placebo (n = 6). The insulin-infused fetal sheep had higher mean +/- SD plasma insulin concentrations (448 +/- 507 versus 11 +/- 8 mU/L; p less than 0.001) and lower arterial oxygen saturations (38 +/- 7 versus 54 +/- 9%; p less than 0.02). The insulin-infused group had a lower mean plasma iron concentration (20.8 +/- 10.9 versus 42.1 +/- 14.7 microM/L; p less than 0.02) and total iron-binding capacity saturation (36 +/- 20 versus 64 +/- 22%; p less than 0.02) and a higher total red cell Hb (45.4 +/- 8.7 versus 32.6 +/- 8.8 g; p less than 0.02) and total red cell iron content (154 +/- 29 versus 111 +/- 29 mg; p less than 0.02) when compared with the placebo group. Seven to 12 d of intrauterine hyperinsulinemia decreases serum iron and increases total red cell iron, most likely by stimulating increased Hb synthesis in response to low arterial oxygen saturation.(ABSTRACT TRUNCATED AT 250 WORDS)

Cerebral hyperperfusion associated with hypertension, may play an important role in the pathogenesis of intraventricular hemorrhage in preterm infants. To examine the effect of hypertension on changes in total and regional brain blood flow (BBF), we increased the mean arterial blood pressure (MABP) in nine awake newborn piglets by an infusion of 0.7 mg/kg of metaraminol bitartrate (Aramine) (group I) and studied cerebral circulatory changes. In order to prevent the Aramine-associated bradycardia, we pretreated nine other piglets with atropine, which produced a higher level of hypertension (group II). MABP and BBF were measured and cerebral vascular resistance (CVR) was calculated during baseline, the Aramine infusion, and twice at decreasing MABP following the discontinuation of the Aramine infusion. In group I, the significant increase in MABP from 68 +/- 3 to 100 +/- 3 mm Hg (mean +/- SEM) during the Aramine infusion resulted in a significant increase in BBF (98 +/- 9 to 118 +/- 11 ml X min-1 X 100 g-1). MABP decreased significantly (although remained significantly above baseline levels), when Aramine was discontinued; however, total BBF remained elevated. CVR increased during the Aramine infusion, but decreased significant (versus the Aramine-infused state) in the post-Aramine period. Regional BBF increased significantly to the cerebrum and cerebellar cortex, but remained unchanged to the other regions including the brain stem. In group II, the Aramine infusion resulted in a significantly greater increase in MABP, a sustained increase in vascular resistance, and no increase in total BBF. Thus, atropine prevents increased BBF during hypertension in the newborn piglet.

Our study tests the hypothesis that hyperviscosity independent of arterial O2 content reduces cerebral cortical blood flow, O2 delivery, and O2 uptake. After baseline determinations, ten 2- to 4-day-old awake spontaneously breathing piglets were given an intravenous infusion (5 ml.kg-1, body weight) of concentrated cryoprecipitate, whereas eight controls received normal saline. Cerebral cortical blood flow, arterial and superior sagittal sinus O2 content, whole blood viscosity, hematocrit, blood gases, and plasma fibrinogen concentrations were measured at baseline and 3 h after infusion. No significant changes were observed in the control group. Three hours after the infusion of concentrated cryoprecipitate the experimental group showed an increase in whole blood viscosity, whereas hematocrit and arterial O2 content were unchanged. There was a decrease in cerebral cortical blood flow and cerebral cortical O2 delivery, whereas cerebral cortical O2 uptake was unchanged. We conclude that hyperviscosity independent of arterial O2 content reduces cerebral cortical blood flow and that although O2 delivery was reduced in the newborn piglet cerebral cortical O2 uptake was maintained.