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Background: Preterm infants show delayed development of motor function after birth. This may relate to functional immaturity of many organs, including the gut and brain. Using pigs as model for preterm infants, we hypothesized that early initiation of enteral feeding stimulates both gut growth and neonatal physical activity., Methods: In experiment 1, preterm and term pigs were fed parenteral nutrition (PN) or PN plus bovine colostrum (BC, 16-64 ml/kg/d enterally) for 5 d. In experiment 2, preterm pigs were fed PN+BC or PN+formula for 5 d. In experiment 3, preterm pigs were fed BC, formula, or human milk (HM) for 10 d. Incubator home cage activity (HCA) was quantified by continuous camera recordings., Results: Preterm birth was associated with reduced intestinal weight and HCA (experiment 1), and BC or formula supplementation increased intestinal weights and HCA (experiments 1+2). Enteral BC and HM feeding increased HCA, intestinal weights, and necrotizing enteritis resistance, relative to formula (experiment 3)., Conclusion: Preterm pigs show decreased physical activity, and the first enteral feeds diet dependently stimulate both gut growth and physical activity. The effects may arise from maturation of digestive, metabolic, and neurological functions, including gut serotonin production, by the first enteral feeds and milk bioactive factors.

Introduction: In preterm neonates, peroxides contaminating total parenteral nutrition (TPN) contribute to oxidative stress, which is suspected to be a strong inducer of hepatic complications related to prematurity. Recently, others reported that hexapeptides derived from human milk (HM) exerted free radical-scavenging activities in vitro. Therefore, the aim of this study was to assess the capacity of these hexapeptides to limit the generation of peroxides in TPN and to prevent TPN-induced hepatic oxidative stress., Methods: At 3 d of life, guinea pigs were infused, through a catheter in jugular vein, with TPN containing or not peptide-A (YGYTGA) or peptide-B (ISELGW). Peroxide concentrations were measured in TPN solutions, whereas glutathione, glutathionyl-1,4-dihydroxynonenal (GS-HNE) and mRNA levels of interleukin-1 (IL-1) and tumor necrosis factor-α (TNFα) were determined in liver after 4 d of infusion., Results: The addition of peptide-A to TPN allowed a reduction in peroxide contamination by half. In vivo, peptide-A or peptide-B corrected the hepatic oxidative status induced by TPN. Indeed, both peptides lowered the hepatic redox potential of glutathione and the level of GS-HNE, a marker of lipid peroxidation. As compared with animals infused with TPN without peptide, the hepatic mRNA levels of IL-1 and TNFα were lower in animals infused with TPN containing peptide-A or peptide-B., Discussion: These results suggest that the addition of YGYTGA or ISELGW to TPN will reduce oxidative stress in newborns. The reduction in mRNA of two proinflammatory cytokines could be important for the incidence of hepatic complications related to TPN.

Introduction: Mild therapeutic hypothermia (HT) reduces brain injury in survivors after perinatal asphyxia. Recent guidelines suggest that resuscitation of term infants should be started with air, but supplemental oxygen is still in use. It is not known whether supplemental oxygen during resuscitation affects the protection offered by subsequent HT., Results: Wilcoxon median (95% confidence interval) hippocampal injury scores (range 0.0-4.0; 0 to ≥90% injury) were 21% O(2) normothermia (NT): 2.00 (1.25-2.50), 21% O(2) HT: 1.00 (0.50-1.50), 100% O(2) NT: 2.50 (1.50-3.25), and 100% O(2) HT: 2.00 (1.25-2.50). Although HT significantly reduced hippocampal injury (B = -0.721, SEM = 0.297, P = 0.018), reoxygenation with 100% O(2) increased injury (B = +0.647, SEM = 0.297, P = 0.033). Regression constant B = 1.896, SEM = 0.257 and normally distributed residuals., Discussion: We confirm an ~50% neuroprotective effect of therapeutic HT in the neonatal rat. Reoxygenation with 100% O(2) increased injury and worsened reflex performance. HT was neuroprotective whether applied after reoxygenation with air or 100% O(2). However, HT after 100% O(2) gave no net neuroprotection., Methods: In an established neonatal rat model, hypoxia-ischemia (HI) was followed by 30-min reoxygenation in either 21% O(2) or 100% O(2) before 5 h of NT (37 °C) or HT (32 °C). The effects of HT and 100% O(2) on histopathologic injury in the hippocampus, basal ganglia, and cortex, and on postural reflex performance 7 d after the insult, were estimated by linear regression.

Introduction: There is little information regarding the comparative hemodynamic effects of adding milrinone or levosimendan to dopamine infusion in hypoxia-reoxygenated (H-R) newborns., Results: Severely hypoxic piglets had cardiogenic shock with depressed cardiac index (CI) and mean arterial pressure (MAP). The hemodynamics deteriorated gradually after initial recovery upon reoxygenation. Heart rate and CI improved with milrinone (D+M) and levosimendan (D+L) administration (P < 0.05 vs. control). Both regimens improved carotid arterial flow and carotid vascular resistance; D+M additionally improved superior mesentric arterial flow (all P < 0.05 vs. control). No effect was found on renal arterial flow or elevated lactate state with either regimen. D+M piglets also had a lower myocardial oxidized/reduced glutathione ratio (P < 0.05 vs. control)., Discussion: In conclusion, adding milrinone or levosimendan to dopamine similarly improved systemic hemodynamics in H-R newborn piglets. Milrinone also improved mesenteric perfusion and attenuated myocardial oxidative stress., Methods: Twenty-eight piglets (1-4 d, 1.5-2.5 kg) were instrumented for continuous monitoring of systemic MAP and pulmonary arterial pressure (PAP), CI, and carotid, superior mesenteric, and renal arterial flows. Piglets were randomized with blinding to sham-operated, H-R control (saline), and H-R dopamine (10 μg/kg/min) with D+M or D+L groups. H-R piglets underwent H-R followed by 2 h of drug infusion after reoxygenation. Tissue was collected for biochemical/oxidative stress testing and histological analysis.

The developmentally regulated hemodynamic effects of vasoactive medications have not been well characterized. We used traditional and near-infrared spectroscopy monitoring technologies and investigated the changes in heart rate, blood pressure, common carotid artery (CCA) blood flow (BF), cerebral, renal, intestinal, and muscle regional tissue O2 saturation, and acid-base and electrolyte status in response to escalating doses of vasoactive medications in normotensive anesthetized neonatal piglets. We used regional tissue O2 saturation and CCA BF as surrogates of organ and systemic BF, respectively, and controlled minute ventilation and oxygenation. Low to medium doses of dopamine, epinephrine, dobutamine, and norepinephrine increased blood pressure and systemic and regional BF in a drug-specific manner, whereas milrinone exerted minimal effects. At higher doses, dopamine, epinephrine, and norepinephrine but not dobutamine decreased systemic, renal, intestinal, and muscle BF, while cerebral BF remained unchanged. Epinephrine induced significant increases in muscle BF and serum glucose and lactate concentrations. The findings reveal novel drug- and dose-specific differences in the hemodynamic response to escalating doses of vasoactive medications in the neonatal cardiovascular system and provide information for future clinical studies investigating the use of vasoactive medications for the treatment of neonatal cardiovascular compromise.

Complications after cardiac surgery in neonates can occur because of activation of the inflammatory system. This study used lipopolysaccharide (LPS) endotoxin exposure to cause cytokine activation in neonatal mice and examine left ventricular (LV) function and the effects of antioxidant treatment on cytokine levels. Neonatal mice (6 d old) were injected with either 25 mg/kg LPS (n = 13) or PBS (n = 14), and LV function (echocardiography) was measured at 4 h. Plasma levels of TNF-α, IL-4, IL-6, and IL-10 were measured at 30 min, 1, 2, and 4 h after injection (n = 5 mice per group). Effects of pretreatment with N-acetylcysteine (NAC, 50 mg/kg) on cytokine levels were examined at 2 and 4 h after PBS or LPS (n = 5 mice per group). Four hours after LPS, heart rate was increased (434 ± 14 versus 405 ± 14 bpm, p < 0.05). LV end-diastolic dimension and ejection time were reduced with LPS (both p < 0.05). LPS exposure increased plasma TNF-α, IL-6, and IL-10 levels. NAC pretreatment attenuated the increases in TNF-α and IL-6 levels, but augmented IL-10 levels at 2 h post-LPS. LPS exposure altered cardiac performance and activated cytokines in neonatal mice, which may be ameliorated using antioxidants.

Repeated pain during brain development can have long-term consequences in both humans and animals. We previously showed that maternal care provided to pups experiencing pain reduced adult pain sensitivity. This study tested whether sensory stimulation was responsible for this effect. Rat pups were either mother-reared controls (MR-CON) or artificially reared (AR) with minimal (AR-MIN) or maximal (AR-MAX) stimulation provided daily. In each rearing condition, pups were either uninjected or injected from postnatal day (PND) 4 to 14 with saline (0.9%) or formalin (0.2-0.4%). Pain behavior and paw inflammation were scored. Thermal sensitivity and responses to formalin were tested in adulthood (PND 70). AR neonates, irrespective of sensory stimulation received, exhibited a pain response (p < 0.001), even with a mild formalin dose. Maternal rearing reduced inflammation during the second week of life compared with AR pups (p < 0.05). Early pain exposure did not modify adult pain sensitivity. However, rearing altered adult pain sensitivity such that uninjected MR-CON rats had lower pain sensitivities than uninjected AR rats (p < 0.05). This suggests that the beneficial effects of maternal rearing can be obliterated if additional stimulation/stress occurs during the early neonatal period. In addition, this suggests that optimal level of maternal stimulation exists that determines adult pain sensitivity.

Fetal or early postnatal stressors may predispose infants to develop diabetes, metabolic syndrome, or stroke. We hypothesized that postnatal stress will predispose animals to develop metabolic syndrome and impair the physiologic response to hypoxic-ischemic brain injury. We characterized the short- and long-term physiologic responses to postnatal stress by examining corticosterone (CS), glucose metabolism, and brain injury in neonatal and adult rats exposed to hypoxia-ischemia (H-I). Rat pups were divided into three levels of postnatal stress from postnatal day (P) 3 to P7. All rats underwent unilateral brain injury on either P7 or P134. We measured brain injury, growth, blood pressure, urine/plasma CS, plasma leptin, insulin, and glucose before and after H-I. Postnatal stress increased neonatal CS production, exacerbated neonatal white matter injury, and was associated with adult hyperglycemia after H-I despite increased insulin production. There were no group differences in adult weight, blood pressure, or leptin. Postnatal stress exacerbated brain injury and produced adult hyperglycemia, triggered after hypoxia exposure, consistent with the hypotheses that neonates exposed to early stress are more vulnerable to hypoxia and may be predisposed to develop metabolic syndrome in adulthood. Prolonged maternal separation produced more hyperglycemia than did brief daily handling.

The effect of inflation length on lung aeration pattern, tidal volumes, and functional residual capacity (FRC) immediately after birth was investigated. Preterm rabbits (28 d), randomized into four groups, received a 1-, 5-, 10-, or 20-s inflation (SI) followed by ventilation with 5 cm H2O end-expiratory pressure. Gas volumes were measured by plethysmography and uniformity of lung aeration by phase contrast x-ray imaging for 7 min. The first inspiratory volume significantly (p < 0.001) increased with inflation duration from a median (IQR) of 0.2 (0.1-3.1) mL/kg for 1-s inflation to 23.4 (19.3-30.4) mL/kg for 20-s SI. The lung was uniformly aerated, and the FRC and tidal volume fully recruited after 20-s SI. A 10-s SI caused a higher FRC (p < 0.05) at 7 min, and a 20-s SI caused a higher FRC (p < 0.05) at 20 s and 7 min than a 1- or 5-s SI. The mean (SD) time for 90% of the lung to aerate was 14.0 (4.1) s using 35 cm H2O peak inflation pressure. In these rabbits, 10- and 20-s SI increased the inspiratory volume and produced a greater FRC, and a 20-s SI uniformly aerated the lung before ventilation started.

The lungs of very preterm infants have immature airways and gas exchange structures and are usually surfactant deficient. Antenatal corticosteroids are commonly used to enhance fetal lung maturation in preterm infants, but little is known of their effects on pulmonary blood flow (PBF) before and immediately after birth. Our aim was to determine the effects of antenatal betamethasone on PBF before birth and during the postnatal transition in very preterm lambs. Antenatal betamethasone treatment significantly increased mean fetal PBF from 20.2 +/- 5.1 to 84.3 +/- 18.3 mL/min at 30 h after administration; the PBF waveform was also significantly altered. Mean diastolic PBF increased from -38.5 +/- 4.9 pretreatment to -10.2 +/- 11.0 mL/min at approximately 36 h after the initial betamethasone dose (negative values indicate retrograde flow away from the lungs). Within 10 min after delivery, PBF was similar in control and betamethasone-treated lambs. These data demonstrate that antenatal betamethasone significantly increases fetal PBF and alters the PBF waveform but has little effect on postnatal PBF.

Pulmonary deposition of inhaled drugs in ventilated neonates has not been studied in vivo. The objective of this study was to evaluate pulmonary delivery of gadopentetate dimeglumine (Gd-DTPA) following nebulization in ventilated piglets using magnetic resonance imaging. Seven ventilated piglets (5 +/- 2 d old, weight 1.8 +/- 0.5 kg) were scanned in the Bruker/Siemens 4T magnetic resonance scanner using T1 weighted spin-echo sequence. Aerosols of Gd-DTPA were generated continuously using the MiniHeart jet nebulizer. Breath-hold coronal images were obtained before and every 10 min during aerosolized Gd-DTPA for 90 min. Signal intensity (SI) changes over the lungs, kidneys, liver, skeletal muscle, and heart were evaluated. A significant increase in SI was observed in the lungs, kidney, and liver at 10, 20, and 40 min respectively after start of aerosol. At the end of 90 min, the SI increased by 95%, 101%, and 426% over the right lung, left lung, and kidney, respectively. A much smaller increase in SI was observed over the liver. In conclusion, we have demonstrated effective pulmonary aerosol delivery within 10 min of contrast nebulization in ventilated piglets. Contrast visualization in the kidneys within 20 min of aerosol initiation reflects alveolar absorption, glomerular filtration and renal concentration.

Suboptimal developmental environments program offspring to lifelong metabolic problems. We evaluated effects of maternal isocaloric low protein diet during pregnancy and/or lactation on milk quantity and leptin concentration at postnatal day 7, 14, and 21. Control mothers ate 20% casein (C) and restricted mothers (R) 10% casein to provide four groups: CC, RR, CR, and RC (first letter pregnancy and second lactation diet) to enable evaluation of effects influenced by maternal diet during pregnancy and lactation. Milk leptin was not a determinant of pup serum leptin. Pup serum leptin did not inhibit milk appetite at any postnatal age. Pup serum leptin did not correlate with pup adipose tissue. Finally, the normal postnatal leptin rise in pup serum was delayed by prenatal undernutrition. These data suggest that fetal nutrition modifies timing of neonatal leptin surge and may contribute to the development of altered appetite and metabolic disorders in later life.

Study investigated neuroutcome in mice subjected at 7-8 d of life to hypoxic-ischemic brain injury (HI) followed by 30 min of reoxygenation with 100% O(2) (Re-O(2)) or room air (Re-Air). At 24 h of recovery, mouse reflexes were tested. At 7 wks after HI spatial orientation and memory were assessed in the same mice. Mortality rate was recorded at 24 h and at 7 wks of recovery. In separate cohort of mice, changes in cerebral blood flow (CBF) during HI-insult and reoxygenation were recorded. Re-O(2)versus Re-Air mice exhibited significantly delayed geotaxis reflex. Adult Re-O(2)versus Re-Air mice exhibited significantly better spatial learning and orientation with strong tendency toward better preserved memory. Histopathology revealed significantly less hippocampal atrophy in Re-O(2)versus Re-Air mice. Following a hypoxia-induced hypoperfusion, Re-O(2) re-established CBF in the ipsilateral side to the prehypoxic level significantly faster than Re-Air. The mortality was higher among Re-O2 versus Re-Air mice, although, it did not reach statistical significance. Re-O(2)versus Re-Air restores CBF significantly faster and results in better late neuroutcome. However, greater early motor deficit and higher mortality rate among Re-O(2)versus Re-Air mice suggest that Re-O(2) may be deleterious at the early stage of recovery.

Many infants recovering from acute lung disease and pulmonary hypertension still have evidence of reactive airways disease at one year of age, suggesting longer-term airway effects. We hypothesized that parallel changes in smooth muscle would occur in airways and pulmonary arteries from animals with pulmonary hypertension and during normoxic recovery. Thus, two-hour-old piglets were subjected to 3 d chronic hypobaric hypoxia and 3-d-old piglets were subjected to 11 d hypoxia. Some animals were allowed to recover in room air for 3 or 6 d. The amount of smooth muscle and responses of isolated paired bronchial and pulmonary artery rings to endothelin-1 (ET-1) and norepinephrine were studied at the end of hypoxic exposure, on recovery and in age-matched control animals. In all hypoxia induced pulmonary hypertensive animals, smooth muscle area and ET-1 contractile response was increased in the pulmonary arteries and bronchi. Norepinephrine-induced relaxant response was impaired significantly in both bronchi and pulmonary arteries. After 3 d recovery, pulmonary arterial smooth muscle area decreased by 65%, and ET-1-induced contractile responses were normal for age. In the airways, ET-1 contractile response only normalized after six days and bronchial smooth muscle was still increased. After 6 d recovery pulmonary arterial norepinephrine-induced relaxant response had returned to normal, but bronchial response remained impaired. Thus during pulmonary hypertension, both bronchial and pulmonary arterial smooth muscle area and contractile responses are increased. On recovery, regression of bronchial structural and functional abnormalities is slower than in pulmonary arteries.

Early disturbances in breathing control, including apneas of prematurity and apparently life-threatening events, account for some cases of sudden infant death syndrome and for a rare disorder called congenital central hypoventilation syndrome (CCHS). Data suggesting a genetic basis for CCHS have been obtained. Recently, we found heterozygous de novo mutations of the PHOX2B gene in 18 of 29 individuals with CCHS. Most mutations consisted of five to nine alanine expansions within a 20-residue polyalanine tract, probably resulting from nonhomologous recombination. Other mutations, generally inherited from one of the parents, in the coding regions of genes involved in the endothelin and RET signaling pathways and in the brain-derived-neurotrophic factor (BDNF) gene have been found in a few CCHS patients. Interestingly, all these genes are involved in the development of neural crest cells. Targeted disruption of these genes in mice has provided information on the pathophysiological mechanisms underlying CCHS. Despite the identification of these genes involved in breathing control, none of the genetically engineered mice developed to date replicate the full human CCHS respiratory phenotype. Recent insights into the genetic basis for CCHS may shed light on the genetics of other early disturbances in breathing control, such as apnea of prematurity and sudden infant death syndrome.

Severely premature infants are often at increased risk of cerebral hemorrhage and/or ischemic injury caused by immature autoregulatory control of blood flow to the brain. If blood flow is too high, the infant is at risk of hemorrhage, whereas too little blood flow can result in ischemic injury. The development of a noninvasive, bedside means of measuring cerebral hemodynamics would greatly facilitate both diagnosis and monitoring of afflicted individuals. It is to this end that we have developed a near infrared spectroscopy (NIRS) system that allows for quantitative, bedside measurement of cerebral blood flow (CBF), cerebral blood volume (CBV), and mean transit time (MTT). The technique requires an i.v. injection of the near infrared chromophore indocyanine green. Six newborn piglets, median age of 18 h (range 6-54 h), median weight of 1.75 kg (range 1.5-2.1 kg), were studied. Measurements of CBF, CBV, and MTT were made at normocapnia, hypocapnia, and hypercapnia to test the technique over a range of hemodynamic conditions. The accuracy of our new approach has been determined by direct comparison with measurements made using a previously validated computed tomography technique. Paired t tests showed no significant difference between computed tomography and NIRS measurements of CBF, CBV, and MTT, and mean biases between the two methods were -2.05 mL x min(-1) x 100 g(-1), -0.18 mL x 100 g(-1), and 0.43 s, respectively. The precision of NIRS CBF, CBV, and MTT measurements, as determined by repeated-measures ANOVA, was 9.71%, 13.05%, and 7.57%, respectively.

After birth, the full-term ductus arteriosus actively constricts and undergoes extensive histologic changes that prevent subsequent reopening. These changes are thought to occur only if a region of intense hypoxia develops within the ductus wall after the initial active constriction. In preterm infants, indomethacin-induced constriction of the ductus is often transient and is followed by reopening. Prostaglandins and nitric oxide both play a role in inhibiting ductus closure in vitro. We hypothesized that combined inhibition of both prostaglandin and nitric oxide production (with indomethacin and N-nitro-L-arginine (L-NA), respectively) may be required to produce the degree of functional closure that is needed to cause intense hypoxia. We used preterm (0.67 gestation) newborn baboons that were mechanically ventilated for 6 d: 6 received indomethacin alone, 7 received indomethacin plus L-NA, and 16 received no treatment (control). Just before necropsy, only 25% of control ductus and 33% of indomethacin-treated ductus were closed on Doppler examination; in contrast, 100% of the indomethacin-plus-L-NA-treated ductus were closed. Control and indomethacin-treated baboons developed negligible-to-mild ductus hypoxia (EF5 technique). Similarly, there was minimal evidence of ductus remodeling. In contrast, indomethacin-plus-L-NA-treated baboons developed intense hypoxia in regions where the ductus was most constricted. The hypoxic muscle strongly expressed vascular endothelial growth factor, and proliferating luminal endothelial cells filled and occluded the lumen. In addition, cells in the most hypoxic regions were undergoing DNA fragmentation. In conclusion, preterm newborns are capable of remodeling their ductus, just like the full-term newborn, if they can reduce their luminal blood flow to a point that produces intense ductus wall hypoxia. Combined prostaglandin and nitric oxide inhibition may be necessary to produce permanent closure of the ductus and prevent reopening in preterm infants.

We measured body temperature continuously using telemetry to determine the development of circadian rhythmicity in neonatal baboons after birth. Twelve fetal baboons (nine males and three females) of known gestational age ranging from 167 to 193 d were studied. We eliminated the influence of maternal factors by hand rearing these infants from the moment of birth until 45 d of life. All infants showed steady growth in body weight, head circumference, and crown-rump length. Neurobehavioral responses including visual and auditory orientation, motor maturity, irritability, and consolability increased as a function of age. Circadian rhythms of body temperature were present in the second week of life, and the amplitude of this rhythm increased throughout the developmental period studied. The increase in the amplitude of circadian body temperature rhythm independent of environmental time cues may indicate the maturation of the brain. These neonatal nonhuman primates offer an excellent model for studying neurobehavioral development and maturation of circadian rhythms while controlling external factors in a manner that is not possible with human neonates.

We hypothesized that the inter-breath variability of the breathing pattern in newborn rats varied with temperature and oxygenation. Breathing pattern was recorded in 4-day-old rats by airflow plethysmography, during normoxia in warm (control) and cold conditions, or during hypoxia (inspired O2 = 10%) in warm or cold conditions, each lasting 15 min. The warm phase (36 degrees C) either preceded or followed the cold (24 degrees C). Time-domain analysis was applied to 500 continuous breaths recorded toward the end of each phase. All parameters describing the breathing pattern (instantaneous ventilation, tidal volume, and inspiratory and expiratory time) had lower variability when the condition differed from control i.e. in cold or hypoxia, with no correlation with the absolute level of ventilation. The difference in variability between warm-normoxia and the other conditions was reduced when cold preceded the warm phase. Gaseous metabolism was increased in cold because of thermogenesis. When the cold preceded the warm phase the increased thermogenesis partly persisted into the warm phase, raising the metabolic level. We conclude that the variability of the breathing pattern in newborn rats 1) does not depend on the absolute level of ventilation, and 2) is reduced by the increased chemical stimuli occurring during cold-hypermetabolism or hypoxia. In normoxia in warm condition metabolic and chemical stimuli are low, and the variability is the highest. The results are in agreement with the clinical observations of a higher incidence of apneic episodes in infants during warm conditions.

We questioned to what extent changes in temperature could affect the newborn's ventilatory inhibition provoked by lung inflation, or Hering-Breüer (HB) inflation reflex. Conscious newborn rats (3-4 d old) were studied in a double chamber airflow plethysmograph at ambient temperatures of 32 degrees C (slightly below their thermoneutrality), 24 degrees C (cold), and 36 degrees C (warm). At these ambient temperatures, the corresponding body temperatures averaged 35.4, 31.0, and 37 degrees C. The HB reflex was triggered by negative body surface pressures of 5 or 10 cm H2O, and quantified as the duration of the expiratory time during the maintained inflation, either in absolute values or in relation to the control expiratory time. In cold the HB reflex decreased to 80%, and in warm it increased to 150%, of the values measured at 32 degrees C. Oxygen consumption, measured by an open flow system, averaged 59, 47, and 29 mL x kg(-1) x min(-1) at, respectively, 24, 32, and 36 degrees C. In cold, the ventilatory response to hypoxia (10% O2) was almost absent, that to hypercapnia (5% CO2) was greater, and that to hypoxia and hypercapnia combined was less than in warm. We conclude that in newborn rats the strength of the vagal inhibition on breathing, evaluated in the form of the HB reflex, is sensitive to temperature, becoming stronger as temperature increases. One contributing factor is the temperature-induced change in metabolic rate, whereas the role of temperature-induced changes in ventilatory chemosensitivity is variable. The strong temperature-dependency of the neonatal HB reflex implies that in newborns exposed to a warm environment breathing is more susceptible to inhibitory inputs.

Rabbit pups were delivered by cesarean section 1 or 2 d before term, or vaginally around term, and then reared in room air or exposed to intermittent or continuous hyperoxia (> 85%) for up to 9 d. Pups were killed at different ages, and lung hyaluronan (HA; microgram/g of dry lung weight) and lung water content, measured as wet/dry lung weight, were determined. Compared with the day of birth, the lung HA concentration did not change significantly on succeeding days in pups kept in air delivered 2 d (-2 d) or 1 d (-1 d) before term, whereas the water content decreased significantly. Continuous exposure to hyperoxia resulted in a significantly raised lung HA concentration 6 d postterm in both -2 d and -1 d pups, and intermittent exposure to hyperoxia resulted in a significantly raised HA concentration 6 d postterm in -1 d pups, compared with the groups exposed to room air. These increases were accompanied by significantly elevated wet/dry lung weight ratios. Microscopic examination revealed significantly increased HA staining scores in alveoli, arterioles, and bronchioli in both hyperoxia-exposed groups of -2 d pups 6 d postterm, and nonsignificantly higher scores in -1 d and vaginally delivered pups of comparable age, compared with the scores at birth. The results indicate that oxygen exposure neonatally may result in an increase in lung HA accompanied by an increase in lung water content. The increase in lung HA concentration in our study may be an effect of oxygen free radicals or of oxygen-induced stimulation of inflammatory mediators.

To analyze newborn cerebrovascular autoregulation, middle cerebral arteries from 3-4-d-old piglets were cannulated, and diameter changes after transmural pressure variation were measured. After an equilibration period at 30 mm Hg, pressure was modified from 10 to 70 mm Hg in 20-mm Hg steps. Segments with endothelium showed vasodilation during pressure decrease and vasoconstriction during pressure increase. In each case the maximum response was about 5% that of the resting diameter. Segments without endothelium responded passively to pressure change. Vasodilation during pressure decrease was reduced by the preferential calcium-activated potassium (KCa) channel blocker, tetraethylammonium (1 mM), and was absent with the nitric oxide (NO) synthase inhibitor NG-nitro-L-arginine methylester (L-NAME, 10 microM). The NO synthase substrate, L-arginine (10 microM), counteracted the dilation blockade caused by L-NAME. The cyclooxygenase inhibitor indomethacin (10 microM) and the endothelin A receptor antagonist BQ-123 (10O microM) eliminated the pressure increase-induced vasoconstriction. The ATP-sensitive potassium channel blocker, glibenclamide (1 microM), and the endothelin B receptor antagonist, BQ-788 (10 nM), did not modify the autoregulatory response. None of these drugs modified the passive changes produced by pressure variations in segments without endothelium. These results suggest that: 1) piglet middle cerebral artery autoregulation is endothelium-dependent; 2) NO and KCa channels are involved in vasodilation during transmural pressure decrease, and 3) endothelin-1, through endothelin A receptors, and prostanoids mediate vasoconstriction during pressure increase.

The antioxidant vitamins ascorbic acid (AA) and alpha-tocopherol (alpha-TP) effectively inhibit oxygen free radical-induced lipid peroxidation. Using a premature baboon model of hyperoxia-induced bronchopulmonary dysplasia (BPD), we measured concentrations of AA, alpha-TP, and conjugated dienes (CD, marker of lipid peroxidation) in four animals (hyperoxic antioxidant group) receiving high dose antioxidant vitamin supplementation (AA, 100 mg x kg x(-1) x d(-1); alpha-TP; 20 mg x kg x(-1) x d(-1)) and one animal receiving standard dose antioxidant vitamin supplementation (AA, 10 mg x kg x(-1) x d(-1); alpha-TP, 1 mg x kg x(-1) x d(-1)). Respiratory and histopathologic data were compared with data from 10 historical animals exposed to hyperoxia (hyperoxic control group) and 11 historical animals treated as required with oxygen (normoxic control group) who had received standard dose antioxidant vitamin supplementation. Compared with standard dose antioxidant vitamin supplementation, high dose antioxidant vitamin supplementation effectively raised AA concentrations in plasma (37 +/- 22 micromol/L and 395 +/- 216 micromol/L, respectively) and tracheal aspirates (62 +/- 35 micromol/L and 286 +/- 205 micromol/L, respectively), and alpha-TP concentrations in plasma (10.1 +/- 2.5 micromol/L and 24.6 +/- 17.5 micromol/L, respectively). However, there was no apparent effect on tracheal aspirate CD concentrations (482 +/- 333 micromol/L and 1050 +/- 1111 micromol/L, respectively), and respiratory parameters in the hyperoxic antioxidant group were comparable to those of the hyperoxic control group but significantly worse than in the normoxic control group. Finally, no protective effect of high dose antioxidant vitamin supplementation was noted at the histopathologic level.

Gradual pressure overload was induced by abdominal aortic constriction in male rats on postnatal d 6 (AC6) or 2 (AC2). At the age of 8 wk, the systemic blood pressure was measured, and the contractile performance of the left ventricle (LV) was assessed after acute ligation of the ascending aorta in open chest anesthetized animals. The LV free wall was used for the determination of collagen concentration and morphometric analysis of cardiac myocytes and capillaries. Aortic constriction resulted in LV hypertrophy, which was more pronounced in AC2 (by 71%) as compared with AC6 (by 34%) groups and correlated closely with the degree of pressure overload (r = 0.88 and 0.80, respectively). The right ventricular weight was increased by 13% in the AC2 group only. Contractile performance of the LV of aortic constricted rats was significantly higher before as well as after the acute load, but the average functional reserve was unchanged in both experimental groups. Although the maximum value of the rate of pressure development increased linearly with the degree of ventricular hypertrophy in the AC6 group (r = 0.82), a negative correlation was observed in the AC2 animals (r = -0.61). The density of myocytes was decreased, and the calculated average myocyte cross-sectional area was increased in aortic constricted rats, but the coronary capillary density and myocardial concentration of collagen remained constant. Thus, in spite of the larger cardiac growth response in the younger age group, the capillary proliferation and collagen formation were proportional to the ventricular hypertrophy. Therefore, the degrees of overload and hypertrophy do not seem to be limiting factors. Pressure overload induced in newborn rats can be a useful model for the study of mechanisms that control either the growth and differentiation of myocardium soon after birth, as well as the transition from compensated to decompensated hypertrophy at later stages.

Synchronized atrial contraction may be much more important in the newborn, who has a faster heart rate and a less compliant ventricle, than in the adult. We therefore investigated the extent by which synchronized atrioventricular contraction contributes to cardiac output and cardiac work in a neonatal circulation, and whether this effect can be fully explained by the Starling mechanism. In neonatal piglets, left ventricular pressure and volume (conductance catheter) were measured during atrial and ventricular pacing. By manipulating preload during atrial pacing, end-systolic pressure and volume, stroke work, and dP/dtmax were compared at the same end-diastolic volume as indices of contractility. Finally, end-diastolic pressure-volume relationships were assessed to investigate the validity of using end-diastolic pressure as an indicator of preload. We found a significant contribution of synchronized atrial contraction; cardiac output increased 27% when pacing mode was switched form ventricular to atrial. The mechanism by which this was achieved is entirely the enhancement of ventricular filling and thus the Starling effect; contractility was unaffected by pacing mode. This large and important effect can be explained by slowed relaxation (compared with the adult ventricle), which impairs passive filling during the ventricular relaxation phase, and makes active filling during atrial contraction more important. In addition, we found that the use of end-diastolic pressure as an indicator of preload, instead of end-diastolic volume, leads to serious misinterpretations, due to not only the nonlinearity of this relationship, but also the possible shifts in this relationship with certain interventions.

To compare the effects of the sustained exposure to inhaled nitric oxide (NO) on pulmonary gas exchange, cardiovascular function and lung mechanics in newborn lambs with pulmonary hypertension induced by tracheal instillation of meconium. Fifteen newborn lambs (<6 d old) were studied in three groups (n = 5): control, and pulmonary hypertension (5 mL x kg[-1] of a 20% meconium solution) with or without inhaled NO (20 ppm) exposure. Heart rate, systemic and pulmonary arterial pressures, arterial pH and blood gases, cardiac output, and pulmonary mechanics were measured. The exposure to inhaled NO in lambs with pulmonary hypertension, induced by experimental meconium aspiration, produced a transient response. There were a transient improvement in gas exchange, a decrease in pulmonary arterial pressure and airway resistance, without changes in cardiovascular profile. The transient and incomplete response to inhaled NO in experimental MAS might be related to the fact that hypoxemia is not only due to pulmonary vasoconstriction but also to parenchymal lung disease. We hypothesize that this poor and transient response could probably be avoided if strategies that increase lung recruitment, were applied before inhaled NO exposure.

Impaired pulmonary mechanics can cause chest wall distortion (CWD) so that work of breathing is dissipated in deforming the rib cage. We hypothesized that respiratory mechanical unloading as a technique of assisted mechanical ventilation would reduce CWD in animals with injured lungs. We studied five piglets and five adult rabbits to test across different ages and chest configurations. As a result of intratracheal meconium instillation, lung compliance decreased from 21 (median; range 17-35) to 9.5 (6.7-14) mL/kPa/kg in rabbits and from 26 (18-31) to 7.9 (4.9-11) in piglets. Airway resistance increased from 5.0 (4.6-6.1) to 6.9 (5.8-7.9) kPa/L/s in rabbits only. Respiratory inductive plethysmography was used to measure the phase shift between the rib cage and abdominal compartment movements and the total compartmental displacement ratio. We aimed at unloading at least three-fourths of lung elastance in all animals and 2.0 kPa/L/s of resistance in rabbits. Elastic unloading decreased the phase shift in all but one animal. It reduced the total compartmental displacement ratio from 1.27 (1.14-3.73) to 1.16 (1.02-1.82) in piglets and from 1.77 (1.45-5.24) to 1.37 (1.11-4.78) in rabbits. The inspiratory rib cage expansion increased, whereas abdominal expansion did not. The tidal esophageal pressure deflection decreased. Tidal volume increased, whereas respiratory rate remained unaffected so that the partial pressure of arterial CO2 decreased. Resistive unloading as an adjunct to elastic unloading further reduced CWD and induced a more rapid, shallower breathing. We conclude that respiratory unloading as a mechanical support to spontaneous breathing reduces CWD. We speculate that the decrease in CWD increases ventilatory efficiency for a given diaphragmatic effort.

Tidal liquid ventilation (TLV) with perfluorochemical fluid (PFC) has been successfully used experimentally for up to 4 h. However, no studies of prolonged TLV have been reported. We hypothesized that full-term newborn lambs can safely and effectively be liquid-ventilated for up to 24 h. To test this hypothesis, 17 lambs were liquid-ventilated; 7 for 4 h, 5 for 12 h, and 5 for 24 h. Arterial blood samples were obtained for PFC uptake, lipid analysis, and blood gas measurements. Tissues were obtained for histologic and biochemical analysis. Arterial blood gas and mean arterial blood pressure were as follows (mean +/- SEM): pH 7.48 +/- 0.04; PaCO2 30.6 +/- 2.8; PaO2 424 +/- 17; mean arterial pressure 76 +/- 16 mm Hg. PFC blood levels increased rapidly to a mean of 5.2 +/- 3.9 microg/mL. PFC tissue levels increased significantly (p < 0.01) from 260 +/- 45 microg/g at 4 h to 400 +/- 140 microg/g at 12 h. There was no further increase in PFC tissue levels by 24 h (456 +/- 181 microg/g). There was a significant difference in PFC concentration as a function of tissue (p < 0.01). Furthermore, there was a significant correlation (r = 0.88; p < 0.01) between the amount of PFC and lipid in blood and tissue. Microscopic examination of the lungs demonstrated no evidence of barotrauma. These data demonstrate that prolonged TLV can be safe and efficacious for up to 24 h in full-term newborn lambs.

Infants dying with bronchopulmonary dysplasia (BPD) demonstrate increased numbers of pulmonary neuroendocrine cells (PNEC). These infants also possess altered airway epithelial and smooth muscle dimensions reminiscent of oxygen-exposed animals. Because the pathogenesis of BPD involves oxygen toxicity, we hypothesized that chronic hyperoxia would induce both airway remodeling and PNEC hyperplasia. To test this theory, we compared the small airway morphology of 21-d-old rats subsequently exposed to 2 wk of > 95% O2 (Ox; n = 12) with that of normoxic controls (Con; n = 12). In paraffin-embedded sections, airways < 1500 microns cut in cross-section were analyzed using light microscopy and image analysis software. The degree of epithelial and smooth muscle hyperplasia was assessed with proliferating cell nuclear antigen (PCNA). PNEC content was assessed via immunohistochemical staining for calcitonin gene-related peptide (CGRP) and the number of solitary PNEC (PNECsol) and PNEC clusters (neuroepithelial bodies, NEB) counted per section. We found that oxygen exposure increased epithelial and smooth muscle wall thickness (epithelium: Con, 12.3 +/- 1.4 versus Ox, 14.8 +/- 1.4 microns, p < 0.05; smooth muscle: Con, 7.0 +/- 1.0 versus Ox, 10.0 +/- 1.0 microns, p < 0.05). The changes in wall dimensions were accompanied by a 20% increase in fractional PCNA labeling of the epithelium but not the smooth muscle. Both PNECsol and NEB number increased in the Ox group (PNECsol Con, 3.6 +/- 2.6 versus Ox, 6.3 +/- 3.1/100 mm epithelium, p < 0.05; NEB Con, 7.1 +/- 4.0 versus 11.9 +/- 3.6/100 mm epithelium, p < 0.05). These findings document an association between hyperoxia, airway remodeling, and PNEC hyperplasia and imply that PNEC products may contribute to the pathogenesis of oxygen-related pulmonary diseases such as BPD.

Eleven open-chest piglets were studied to examine the effects of vascular engorgement on partitioned airway and lung tissue mechanics, and to investigate the role of vagal denervation on lung function during engorgement. Alveolar pressure was measured using alveolar capsules. Pulmonary elastance (EL) and resistance (RL), airway (Raw), and tissue (Vti) resistance were calculated during mechanical ventilation. Acute fluid administration with polygeline (10 mL/kg boluses up to a total of 30 mL/kg) resulted in an increase of RL with increases in both Raw and Vti. Vti rose 2-3-fold in comparison with Raw. To increase left atrial pressure, a balloon catheter was inserted into the left atrium of the heart and inflated with different volumes. The responses of EL, RL, Raw and Vti were comparable to those with acute fluid load. Alterations in pulmonary mechanics were closely correlated to mean pulmonary artery blood pressure. In another six animals the effect of vagotomy on vascular engorgement was studied. Vagotomy did not alter baseline airway or tissue mechanics. Furthermore, vagotomy did not influence the change of pulmonary mechanics to fluid load or increase of left atrial pressure. Our data indicate that in young piglets vascular engorgement causes alterations in airway and lung tissue mechanics that are not dependent on vagal influences.

Catecholamines are a class of neurotransmitters involved in central nervous system autonomic control. Both acute and chronic hypoxia create alterations in ventilation and blood pressure via catecholamine release, although the mechanisms of these alterations are unknown. The enzymes tyrosine hydroxylase (TH) and phenylethanolamine N-methyltransferase (PNMT) catalyze the rate-limiting step in the catecholamine pathway and production of epinephrine, respectively. Both have been colocalized with Fos protein in metabolic mapping studies of the O2-chemosensory pathway of adult and early postnatal rat. Thus, catecholamines are putative neurotransmitters in a subset of second and higher order respiratory neurons. To characterize the effects of prenatal hypoxia on subsequent TH and PNMT gene and protein expression, pregnant rats were placed in moderate hypoxia (10% O2) from gestational d 18 until birth. Northern and Western analyses of dorsal (catecholaminergic/adrenergic cell group 2) and ventral (catecholaminergic/adrenergic cell group 1) medullary tissue of postnatal (P) age P0, P3, P7, P10, and P14 pups were then done to examine changes in TH and PNMT mRNA and protein compared with normoxia-reared controls. Compared with controls, pups exposed to maternal hypoxia during pregnancy had lower levels of TH mRNA and protein at birth in dorsal medulla and higher levels of TH mRNA the first postnatal week in the ventral medulla. Pups that had been hypoxic in utero showed significantly lower levels of PNMT protein during the second postnatal week in dorsal medulla than did controls. Prenatal hypoxia-induced changes in levels of enzymes responsible for catecholamine synthesis may later be manifest as developmental deficiencies in neuronal function. This may compromise responses to acute hypoxic challenges during early postnatal life and contribute to autonomic nervous system disorders of the newborn such as apnea and sudden infant death syndrome.

Antenatal glucocorticoid therapy improves pulmonary function in preterm newborns. We have determined the effect of antenatal glucocorticoid therapy on isoproterenol and prostaglandin (PG) E2-mediated relaxation in preterm ovine pulmonary veins after birth. Ovine fetuses (121 and 126 d of gestation; term = 150 d) received an ultrasound guided intramuscular injection of betamethasone, 0.5 mg/kg, or saline. Lambs were delivered 15 or 48 h later, ventilated for 3 h, and killed. Isolated fourth generation pulmonary veins were suspended in organ chambers filled with modified Krebs-Ringer solution (95% O2, 5% CO2) at 37 degrees C, and their isometric tension was recorded. During contractions to U46619, isoproterenol and PGE2 induced greater relaxations of pulmonary veins of betamethasone-treated lambs than those of control. Forskolin, an activator of adenylate cyclase, caused greater relaxation in veins of betamethasone-treated lambs than in those of controls. A greater relaxation of veins treated with betamethasone than that of control veins also occurred in the presence of isobutylmethylxanthine, an inhibitor of phosphodiesterases. All vessels relaxed similarly to 8-bromo-cAMP, a cell membrane-permeable analog of cAMP. When stimulated with isoproterenol, PGE2, and forskolin, adenylate cyclase activity of crude membrane preparations of pulmonary veins treated with betamethasone was greater than that of controls. These results demonstrate that antenatal betamethasone therapy potentiates isoproterenol and PGE2-mediated relaxation of pulmonary veins of preterm lambs; an enhanced adenylate cyclase activity explain in part the effect of antenatal glucocorticoid therapy on pulmonary veins of preterm lambs.

The reason why some infants with respiratory distress syndrome fail to respond to surfactant, or respond only transiently, is incompletely understood. We hypothesized that resuscitation with large breaths at birth might damage the lungs and blunt the effect of surfactant. Five pairs of lamb siblings were delivered by cesarean section at 127-128 d of gestation. One lamb in each pair was randomly selected to receive six manual inflations of 35-40 mL/kg ("bagging") before the start of mechanical ventilation, a volume roughly corresponding to the inspiratory capacity of lamb lungs after prophylactic surfactant supplementation. Both siblings were given rescue porcine surfactant, 200 mg/kg, at 30 min of age. Blood gases and deflation pressure-volume (P-V) curves of the respiratory system were recorded until the lambs were killed at 4 h. The P-V curves became steeper after surfactant in the control group, but no such effect was seen in those subjected to bagging. At 4 h, inspiratory capacity and maximal deflation compliance were almost three times higher (p < 0.01) in the controls than in the bagged lambs. The latter were also more difficult to ventilate and tended to have less well expanded alveoli and more widespread lung injury in histologic sections. We conclude that a few inflations with volumes that are probably harmless in other circumstances might, when forced into the surfactant-deficient lung immediately at birth, compromise the effect of subsequent surfactant rescue treatment. Our findings challenge current neonatal resuscitation practice of rapidly establishing a normal lung volume by vigorous manual ventilation.

This study investigated the effects of intraventricular/periventricular blood on cerebral cAMP production and cortical cerebrovascular reactivity. Under halothane and N2O anesthesia, 3 mL of either autologous blood or artificial cerebrospinal fluid (CSF) were injected into the left caudate nucleus; volume was adequate to result in extrusion of fluid or blood into the lateral ventricles of 1-2-d-old piglets. Twenty-four hours later, a closed cranial window was implanted over the left parietal cortex. Pial arteriolar responses to vasodilator and vasoconstrictor stimuli were monitored. Before the application of vasoactive agents, cortical periarachioid CSF was collected for cAMP measurement. Pial arteriolar responses to topical application of endothelin-1 (10(-9) and 10(-8) M) and to leukotriene C4 (10(-10) and 10(-9) M) were similar between the two groups. However, pial arteriolar responses to topical application of cAMP-mediated vasodilators, prostaglandin E2 (10(-6) and 10(-5) M), and histamine (10(-6) and 10(-5) M), respectively, were markedly reduced in the blood group when compared with the artificial CSF (control) group. Mean CSF cAMP level in the blood group was significantly lower than the control group (199 +/- 31 versus 1092 +/- 238 fmol/mL, p = 0.0006). We conclude that in newborn pigs intraventricular/periventricular blood results in a marked reduction of CSF cAMP concentration and attenuation of the cerebrovascular responses to cAMP-mediated vasodilators on the cortical surface remote from the site of blood or hematoma.

Mature myocardium utilizes calcium released by the sarcoplasmic reticulum (SR) for cell contraction. Transient exposure of mature myocytes to caffeine is known to directly trigger Ca2+ release from the SR. In contrast, neonatal rabbit heart cells rely on transsarcolemmal Ca2+ influx for tension generation. SR function is decreased in immature heart and appears to play a minimal role as a calcium source. Accordingly, we hypothesized that neonatal rabbit myocytes would not respond to a caffeine pulse. Isolated neonatal and adult myocytes were paced to load the SR with calcium and then exposed to a 1-s pulse of 10 mM caffeine. As previously described, adult myocytes exhibited a brisk contraction in response to caffeine. Unexpectedly, neonatal myocytes also exhibited a similar, brisk response. These caffeine-induced contractions were not dependent on extracellular Ca2+ but were dependent upon the loading of SR Ca2+ stores. When SR Ca2+ stores were depleted by exposure to caffeine, mature myocytes exhibited only small, slow contractions in response to electrical field stimulation. Replenishing the SR Ca2+ stores resulted in normal, brisk contractions. In contrast, electrically stimulated contractions in immature myocytes were largely unaffected by caffeine-induced SR depletion. Thus, although neonatal myocytes are capable of loading and releasing calcium from the SR, such SR calcium release is not normally required for contraction in the developing heart. The minor role of SR Ca2+ release in immature rabbit heart may not result from immaturity of the SR, but rather from an inadequate mechanism to trigger SR calcium release.

The role of an ATP-dependent K+ channel (K(ATP)+) relaxation in the porcine pulmonary vasculature from birth to adulthood was investigated in vitro using levcromakalim on isolated, prostaglandin F2alpha (30 microM)-precontracted conduit arteries (CA), resistance arteries (RA), and veins (PV). Vessels from neonatal pulmonary hypertensive piglets exposed to chronic hypobaric hypoxia (CHH, 51 kPa) for 3 d, either from birth or from 3 d of age were also studied. Levcromakalim relaxed all vessels in a concentration- and glibenclamide-sensitive manner. In normal CA, the maximal extent of relaxation and sensitivity (EC50) increased between birth and 17 d. Endothelium-removal increased EC50 at 17 d only. Indomethacin (10 microM), but not N(G)-monomethyl-L-arginine (L-NMMA) (30 microM), inhibited relaxation in CA from newborn, 3-d-old, and adult animals. In RA, levcromakalim-induced relaxations did not change during development and endothelium-removal attenuated relaxations in 3-d-old but not in adult animals. At both ages in RA, L-NMMA attenuated relaxations and subsequent addition of L-arginine (1 mM) restored them. In PV, maximal relaxation increased between birth and 6 d with no change of EC50. At all ages, relaxation was partially endothelium-dependent and inhibited by L-NMMA (except in the newborn). Indomethacin only attenuated relaxations in veins from 6- and 17-d-old animals. CHH did not influence relaxant responses in CA and PV but decreased EC50 in RA. Thus K(ATP)+ channel activation caused relaxation from birth onward in all vascular segments with varying endothelium dependence. CHH did not affect relaxation in the large vessels and up-regulated those in RA. These findings indicate a possible role for K(ATP)+ channels during normal adaptation and a potential therapeutic role in the management of pulmonary hypertensive newborn infants.

Circulatory changes occur during perinatal life that increase cardiac output and left ventricular contractile reserve. To examine postnatal changes in left ventricular systolic function and ventricular-vascular coupling, piglets underwent cardiac catheterization at 1, 2, 4, and 6 wk of age. We measured end-systolic elastance (Ees), preload-recruitable stroke work, dP/dt(max), the dP/dt(max) end-diastolic volume relation, cardiac index, heart rate, arterial elastance (Ea), and the ratio Ea/Ees, at rest, during isoproterenol infusions (0.05-1.0 microg/kg/min), and after propranolol (1 mg/kg i.v.). Resting heart rate and cardiac index decreased between 1 and 6 wk. In 1 wk olds, resting Ees was at maximum and was unchanged during isoproterenol infusion; isoproterenol increased other contractility indices. Two, 4, and 6 wk olds demonstrated reserve using all contractility indices. Contractile efficiency was not different between ages. In 1 wk olds, Ea decreased during isoproterenol infusion; isoproterenol did not change Ea at 6 wk. Ea/Ees was higher at rest at 6 wk than at 1 wk, and fell significantly on isoproterenol; isoproterenol did not change Ea/Ees at 1 wk. With beta-adrenergic stimulation, 1 wk olds increased cardiac index by increasing heart rate and decreasing afterload, 6 wk olds increased cardiac index by increasing heart rate and contractility; no change in contractile efficiency was found in either group. In summary, contractile reserve is limited at 1 wk when measured by Ees, but other indices demonstrated reserve. Indexed Ea falls in response to beta-adrenergic stimulation in all ages but 6 wk. Ventricular-vascular coupling is optimized at 1 wk even under baseline conditions.

Circulating glucocorticoids play a role during the immediate postnatal period in adapting the neonate to extrauterine life and are also thought to influence tissue development and function in the later postnatal period. In the present study we have used a rat model to test whether birth by cesarean section (C-section), either alone or with an added period of acute anoxia, affects the development profile of basal corticosterone secretion during the first 5 wk of life. Plasma levels of total corticosterone and of corticosteroid-binding globulin were measured at various times after birth in rats born vaginally, by C-section, or by C-section with 15 min of added anoxia. These measures allowed for calculation of levels of free, biologically active, corticosterone. Under all conditions, total corticosterone appeared to accurately reflect levels of free corticosterone. Plasma corticosterone levels measured immediately (< 5 min) after birth were similar in male rat pups born vaginally, by C-section, or by C-section with added anoxia, whereas female pups born by C-section showed a significant increase in free corticosterone at birth, in comparison with vaginally born females. Both male and female animals born by C-section showed a reduction in plasma corticosterone at 1 h (male: 31% of control, p < 0.01; female: 45% of control, p < 0.05) and at 7 d (male: 61% of control, p < 0.01; female: 55% of control, p < 0.05) after birth, in comparison with vaginally born controls. In animals born by C-section with added anoxia, significant reductions in plasma corticosterone were observed for males at 1 h (58% of control; p < 0.05) and for females at 7 d (62% of control; p < 0.05) after birth. At 14 d of age, corticosterone levels were higher in male rats born by C-section either with (227% of control; p < 0.05) or without (239% of control; p < 0.05) added anoxia, in comparison with vaginally born controls. Thus C-section birth produces an early rise in plasma corticosterone on d 14 away from the low values associated with the adrenal quiescent period in the first 1-2 wk in the rat. By 35 d of age, there were no differences in plasma corticosterone attributable to C-section birth and/or acute birth anoxia, in either male or female rats. It is concluded that, in a rat model, birth by C-section has significant effects on the profile of plasma corticosterone during the early weeks of development, a period though to be critical for effects of corticosteroids on developing tissues. Because the rat at birth is developmentally less mature than is the term human neonate, these findings may have implications for development of the premature human neonate.

It is not known how the mode of flow alters the resistance properties of the newborn pulmonary circulation. Therefore, we compared the pressure responses of the perfused left pulmonary circulation of 16 newborn lambs with step changes in either pulsatile or steady flow. The slope of the pressure-flow curve differed significantly according to mode of perfusion. The slope of the pressure-flow curve was 0.125 +/- 0.115 mm Hg.mL-1.kg.min for pulsatile flow and 0.484 +/- 0.350 mm Hg.mL-1.kg.min for steady flow perfusion (p less than 0.05). The intercept pressure at zero flow was similar for both modes of flow, being 21.3 +/- 4.9 mm Hg for pulsatile flow compared with 23.8 +/- 11.3 mm Hg for steady flow perfusion. All perfusions with either mode of flow demonstrated flow-dependent decreases in pulmonary vascular resistance. However, the decreases in resistance were greater for pulsatile than for steady flow perfusion. In addition, the steady-perfused lungs demonstrated a significantly greater left lung wet/dry weight ratio than either the left lungs of pulsatile-perfused animals or the intact left lungs of 13 control animals not undergoing experimental perfusion (p less than 0.05). Thus, the mode of flow appears to play a major role in modulating the magnitude and distribution of pulmonary vascular resistance. This factor must be considered when interpreting the physiologic significance of hemodynamic experiments.

Single-pulse administration of either recombinant human granulocyte-monocyte colony stimulating factor or recombinant human granulocyte colony stimulating factor to newborn rats has previously been demonstrated to increase the peripheral neutrophil count and modulate bone marrow (BM) neutrophil pools. In our present study, we investigated the effects of 7 d of either recombinant murine granulocyte-monocyte colony stimulating factor (rmGM-CSF) (75 micrograms/kg/d) or recombinant murine IL-3 (rm IL-3) (10 micrograms/kg/d) on newborn rat myelopoiesis. Sprague Dawley newborn rats (greater than or equal to 24 h) were injected (intraperitoneally) daily for 7 d with either rmGM-CSF, rmIL-3, or PBS/BSA. rmGM-CSF induced a significant increase in the peripheral neutrophil count on d 3 (p less than 0.03) and d 7 (p less than 0.001) (75% increase). Additionally, rmGM-CSF induced a 50% increase in the BM neutrophil storage pool (p less than 0.025). rmIL-3 increased the BM colony forming unit-granulocyte monocyte pool (p less than 0.001); however, it failed to increase the peripheral neutrophil count or BM neutrophil storage pool. Neither CSF increased the BM neutrophil proliferative pool or BM colony forming unit-granulocyte monocyte proliferative rate. Additionally, 7 d of rmGM-CSF with or without antibiotics did not synergistically alter the mortality rate after group B streptococcol inoculation. This study suggests that rmIL-3 appears to stimulate more neonatal myeloid committed progenitor cell activity compared with rmGM-CSF. Optimal modulation of neonatal myelopoiesis may require the use of a sequential combination of hematopoietic CSF, namely an early-acting CSF followed by a more lineage myeloid-specific CSF.

Cerebral ischemia in the neonate is an important cause of hypoxic-ischemic encephalopathy. Thus, it is important to have an economical and readily available animal model in which to study the local control of the cerebral circulation in the perinatal period. This study demonstrates that the newborn rabbit, a rather immature species at birth, is a suitable neonatal model in which to measure local cerebral blood flow with quantitative iodo[14C]antipyrine autoradiography. One or 2 d after birth, local cerebral blood flow in the newborn rabbit is low, but flow varies distinctly between regions [e.g. 8.9 +/- 1.5 mL.kg-1.s-1 (53 mL.100 g-1.min-1) in the nucleus tractus solitarius and 3.4 +/- 0.7 mL.kg-1.s-1 (20 mL.100 g-1.min-1) in the frontal cortex]. During early postnatal development (i.e between 1 and 8 d), local cerebral blood flow does not change greatly. However, by 17 d, 22 of 26 brain regions exhibit significant marked increases (200-350%) in local cerebral blood flow when compared with blood flow in the newborn. Between 17 and 40 d postnatally, cerebral blood flow continues to increase in 16 of 26 regions (e.g. thalamic areas). In four of the cerebral cortical areas, elevations in flow continue during the period between 40 d of age and adulthood. In contrast to the generalized increases in flow occurring postnatally, a few brain regions (i.e. within the pons and medulla) exhibit only minor changes in cerebral blood flow. The differential pattern and lower basal levels of cerebral blood flow in the neonate compared with the adult may be important determinants in regional susceptibility of the brain to ischemia.

Indices of global systolic performance of the newborn left ventricle exceed those of the adult, despite isolated tissue studies showing immature contractile mechanisms. To evaluate contractility in situ, we investigated the end-systolic pressure-volume relationship (ESPVR) by the conductance technique in nine newborn lambs. After percutaneous placement of catheters, we generated ESPVR by inferior vena cava occlusion, aortic occlusion, and volume infusion in two control states, during three levels of dobutamine infusion, and after propranolol. We performed linear and nonlinear regression analyses of the end-systolic points and derived the slope (Ees) and volume at 14 kPa pressure. We found that reliable ESPVR could be obtained in almost all inferior vena cava and aortic occlusions (50 of 51 in each), but in only 18 of 27 volume infusions. Overall, linear regressions adequately defined the ESPVR (75 of 102 were not statistically different than nonlinear regressions; of those different, the mean linear R2 was 0.934 +/- 0.048). By multiple regression analysis, neither Ees nor volume at 14 kPa significantly changed with dobutamine, but both changed after propranolol (23% less than control and 54% greater, respectively), supporting previous studies showing a limited contractile reserve in the newborn secondary to high resting beta-adrenergic tone. Neither Ees nor volume at 14 kPa was different between control states. However, Ees was 25% less steep when generated by inferior vena cava than by aortic occlusion. We conclude that the ESPVR can be generated reliably and reproducibly in the newborn lamb and is relatively linear and sensitive to changes in contractility, but that it is also sensitive to the technique of load intervention.

To assess the short-term effects of high-frequency jet ventilation (HFJV) on the neonatal lung, 28 newborn piglets were studied. Nine piglets were unventilated except during brief pulmonary measurements, nine animals were conventionally ventilated (arterial CO2 tension 35-45 torr, arterial O2 tension 70-80 torr) for 4 h, and 10 piglets were ventilated with HFJV for the same period. Pulmonary function was analyzed using a computerized technique and tracheobronchial aspirates were examined for biochemical indicators of lung injury; after 4 h, bronchoalveolar lavage was obtained for surfactant composition and activity, and lung sections were examined by light and electron microscopy. Results showed that HFJV provided adequate ventilation at lower inspiratory pressure compared with conventional ventilation (8.6 +/- 0.3 versus 13.8 +/- 1.3 cm H2O; p less than 0.01), while pulmonary mechanics did not vary significantly among the three animal groups. Tracheobronchial aspirates from HFJV animals had higher elastase activity versus unventilated piglets (118.5 +/- 14.1 versus 57.7 +/- 8.4 micrograms/mL; p less than 0.01), as well as higher albumin concentration versus unventilated animals (94.2 +/- 18.7 versus 23.2 +/- 6.5 micrograms/mL; p less than 0.01). In addition, there were small but statistically significant differences between all three groups in the distribution of surfactant phospholipids in bronchoalveolar lavage, although biophysical activity was normal. Scanning electron microscopy revealed flattening of Clara cells in the terminal bronchioles of HFJV animals due to loss of glycogen and secretory granules. These data indicate that despite lower peak inspiratory pressures, HFJV can cause subtle biochemical changes in lungs. Further studies are indicated to determine if these changes precede significant lung injury.

We studied the role of the autonomic nervous system in the regulation of heart rate variation (HRV) in 12 chronically instrumented neonatal lambs. HRV was quantified from ECG tracings by computing periodic HRV distributions at frequencies of 0.02-1.00 Hz, using power spectral analysis of heart rate, and also by HRV indices. Heart rate declined more during the 1st than the 2nd mo after birth. Multiple regression analysis showed that the heart rate responses to vagal and to beta-adrenergic blockade had an independent negative association both with age and with the initial mean heart rate, whereas the overall HRV response had a positive association with age. Vagal blockade led to a 70-80% decrease in the beat-to-beat HRV in all lambs (p less than 0.001). The overall HRV indices decreased by 40-65% in lambs (less than 30 d old (p less than 0.001) and about 30% in those greater than 30 d old (p less than 0.05). In the power spectrum the greatest decrease was seen in the high-frequency components of HRV. beta-Blockade led to a decrease of about 50% in all HRV (p less than 0.001) in the younger lambs, without frequency selection. In the older lambs, it had no effect on the beat-to-beat HRV, but the overall HRV (coefficient of variance) decreased maximally by 40% (p less than 0.01), with a significant reduction in the low-frequency components of HRV. These results suggest that in the regulation of HRV after birth dual control via the autonomic nervous system is most important. In the older lambs, developmental changes result in precise regulation of the fast heart rate fluctuations mainly by the vagal division, whereas the slow fluctuations are partially regulated by the vagal and beta-adrenergic divisions.

The impact of early postnatal protein-energy malnutrition and of 4, 7, and 14 d of nutritional rehabilitation on small intestinal growth, development, structure and function was examined in 28-, 32-, 35-, and 42-d-old infant rabbits. Malnutrition was induced by litter expansion 7 d postpartum and, in randomly selected malnourished animals, refeeding was begun at weaning, 28 d. Results are compared to ad libitum fed dietary controls. Malnutrition altered the small intestine of the developing rabbit, as evidenced by: 1) reduced jejunal and ileal mass as shown by decreased mucosal wt, protein, and DNA content; 2) depressed epithelial proliferation and enterocyte migration along the crypt-villus axis; 3) delayed epithelial maturation as measured by mucosal enzyme activities; and 4) enhanced glucose-stimulated Na+ transport. Refeeding stimulated rapid and complete recovery, as evidenced by: 1) restoration of jejunal and ileal mucosal mass within 4 d; 2) enhancement of epithelial renewal and enterocyte migration by 7 d; and 3) complete return of the normal pattern of mucosal enzymes by 14 d. With 7 d of refeeding, glucose-stimulated Na+ transport was down-regulated to the level of dietary controls. We conclude that early postnatal protein-energy malnutrition has a severe impact on small intestinal growth, development, structure, and function. Furthermore, a brief period refeeding induced a rapid and complete recovery of these parameters.

Twelve newborn lambs less than 48 hr old had their high oxygen affinity blood exchanged for low-affinity fresh adult blood. Tissue oxygenation, hemodynamic status, blood gases, and myocardial function were compared before and after the exchange transfusion. The P50 was increased from 18 to 29 mm Hg after exchange transfusion; pH and hemoglobin levels remained constant; and there was no change in myocardial function, arteriovenous O2 content difference, O2 consumption, or cardiac output. However, mixed venous PO2 was significantly increased with low affinity red cells (33 versus 23 mm Hg).

Group B streptococcal sepsis in newborns produces pulmonary arterial hypertension and hypoxemia. The purpose of this study was to investigate the mechanisms by which hypoxemia occurs. Ten anesthetized, ventilated piglets were infused with 2 X 10(9) colony forming units/kg of Group B streptococci over a 30-min period. Pulmonary arterial pressure rose from 14 +/- 2.8 to 38 +/- 6.7 torr after 20 min of the bacterial infusion (p less than 0.01). During the same period, cardiac output fell from 295 to 184 ml/kg/min (p less than 0.02). Arterial PO2 declined from 97 +/- 7 to 56 +/- 11 torr (p less than 0.02) and mixed venous PO2 fell from 39.6 +/- 5 to 28 +/- 8 torr (p less than 0.05). The multiple inert gas elimination technique was used to detect increases in shunt and alterations in ventilation-perfusion matching. Intrapulmonary shunt did not increase during or after the infusion with group B streptococci. However, there was a significant increase (p less than 0.05) in the SD of pulmonary blood flow, an index of VA/Q mismatching, 20 min after initiation of the infusion of bacteria. All the above changes reverted toward baseline during the 2-h period following discontinuation of the infusion. We conclude that the hypoxemia occurring in the early phase of group B streptococcal sepsis does not develop solely because of increased shunt, but rather is produced by a decline in cardiac output in conjunction with mismatching of pulmonary perfusion to alveolar ventilation.

The developmental response to the euglycemic hyperinsulinemic clamp was assessed among newborn and adult beagles to investigate neonatal insulin resistance. Both neonatal dogs and adults were clamped at euglycemic blood glucose concentrations while receiving insulin at a rate of 3.75, 15, 30, 60, 75 or 100 mU/kg/min to generate a dose-response curve. Blood glucose levels, plasma insulin concentrations and glucose turnover rates during the basal preclamp fasting period were similar in pups and adult dogs. During the clamp period, blood glucose levels were equivalent to the preclamp period whereas plasma insulin concentrations increased. Total glucose utilization increased from 28.7 +/- 18 to a plateau level of 114 +/- 52.3 mumol/kg/min in adult dogs and from 30.9 +/- 10.9 to 53.9 +/- 28.9 mumol/kg/min in newborn dogs. Suppression of endogenous glucose production during euglycemic hyperinsulinemia was close to 100% among adult dogs, whereas in the newborn pups, endogenous glucose production was suppressed 80%. To determine if the newborn was unable to increase glucose uptake because of a saturation effect on glucose utilization independent of insulin, another group of adult and newborn dogs received a hyperglycemic hyperinsulinemic clamp. In response to the hyperglycemic clamp, adults demonstrated an increase of glucose utilization to 261 +/- 83, and newborn dogs increased utilization to 227 +/- 76 mumol/kg/min.(ABSTRACT TRUNCATED AT 250 WORDS)