Thomas H. Pauly

In utero nicotine exposure causes persistent, gender-dependant changes in locomotor activity and sensitivity to nicotine in C57Bl/6 mice.

Maternal cigarette smoking during pregnancy can result in a wide variety of adverse fetal outcomes, ranging from preterm delivery and low birth weight, to sudden infant death syndrome. In addition, in utero tobacco smoke exposure is associated with delayed or impaired neuropsychological development. Although the causative agent in tobacco smoke that leads to these aberrations is not known, some studies have concluded that nicotine may play an important role. Many studies using animal models of prenatal nicotine exposure have supported the hypothesis that nicotine may directly and/or indirectly cause impairments in fetal and neonatal development. However, in many of the animal studies nicotine has been administered acutely to naive dams, which could lead to significant fetal hypoxia; some routes of drug administration are also very stressful to pregnant dams, and changes in stress hormones could also create an unfavorable fetal environment. In this study, pregnant mice were exposed to chronic nicotine via the drinking solution; locomotor activity and sensitivity to nicotine were evaluated in the offspring. We have previously shown that oral nicotine administration produces behavioral and physiological changes that resemble those seen following other routes of nicotine administration. Although oral nicotine exposure did not significantly alter any aspect of the pregnancy, dams drinking a nicotine‐containing solution consumed approximately 20% less volume, compared to saccharin controls. All animals were cross fostered to nicotine naı̈ve lactating dams, immediately after birth. On PN40 and PN60, male mice exposed to in utero nicotine demonstrated significant locomotor hyperactivity in an open filed arena. Although female animals did not show any signs of hyperactivity, they did have a significant attenuation of their hypothermic response to acute nicotine challenge. These results suggest that oral nicotine delivery to pregnant mice causes persistent, gender‐dependant changes in behavior and sensitivity to nicotine. This model may be very useful for future studies that try to more accurately define the windows of sensitivity for nicotine exposure and the possible underlying neurochemical mechanisms involved.

Organ-Specific Disposition of Group B Streptococci in Piglets: Evidence for a Direct Interaction with Target Cells in the Pulmonary Circulation

ABSTRACT: Despite the serious pulmonary manifestations of early onset group B streptococcal (GBS) sepsis, it is not known whether the organism distributes into lung tissue and whether adverse pulmonary hemodynamic abnormalities relate to an interaction between the organism and target cells in the pulmonary vascular bed. Accordingly, this study evaluated the distribution and fate of GBS in the lung, liver, and spleen of anesthetized infant piglets and in isolated, salt solution-perfused piglet lung preparations. GBS were radiolabeled with 111Indium-oxine and infused at a dose of 108 organisms/kg/min for 15 min into anesthetized piglets ranging in age from 5-10 d. Forty-five min after termination of the infusion, animals were killed and specimens of lung, liver, spleen, and blood were excised and the relative deposition and viability of GBS were determined. Most of the recovered bacteria were detected in the lung (53.2 ± 3.9%) followed by the liver (41.4 ± 2.0%) and spleen (2.2 ± 0.38%). GBS detected in the blood was estimated to be only 3.2 ± 1.0% of the infused dose. Viability of GBS was least in the lung (21.4 ± 2.6%) relative to the liver (45.7 ± 11.2%) and spleen (83.4 ± 19.5%). After a 60-min GBS infusion, transmission electron microscopy localized the organism within pulmonary intravascular macrophages in the lung; there was no evidence for bacterial interaction with either neutrophils or endothelial cells. In the liver, GBS was found exclusively in Kupffer cells. In isolated piglet lungs perfused at a constant flow rate with blood-free physiologic salt solution, GBS (106 to 108 organisms/mL) provoked concentration-dependent increases in pulmonary vascular resistance. Transmission electron microscopic examination of isolated lungs indicated that GBS was localized within pulmonary intravascular macrophages, again with no apparent intractions between the organism and other cellular residents of the pulmonary vascular bed. These results indicate that GBS distributes into lung and liver where resident intravascular phagocytes, possibly pulmonary intravascular macrophages and Kupffer cells, respectively, contribute to killing of the organism. In addition, because GBS evokes pulmonary hypertension in isolated piglet lung preparations, it appears that cardiopulmonary disturbances in the intact animal could be initiated by a direct interaction between GBS and resident lung cells without obligatory participation by other organ systems.

Group B Streptococcus Promotes Oxygen Radical-Dependent Thromboxane Accumulation in Young Piglets

ABSTRACT: Both thromboxane A2 and oxygen-derived free radicals appear to play central roles in group B streptococcus (GBS)-induced pulmonary hypertension in piglets. This study tested the hypothesis that GBS promotes oxygen radical-dependent thromboxane accumulation and pulmonary hypertension in infant piglets. Piglets 4-12 d old were anesthetized and prepared for assessment of pulmonary arterial pressure and arterial blood gases. In control animals, GBS (108 organisms/kg/min for 15 min) increased mean pulmonary artery pressure by 30 ± 1.5 torr and reduced arterial PO2 by 100 ± 20 torr. Thromboxane A2, radioimmunoassayed in venous blood as thromboxane B2, increased by 2452 ± 800 pg/mL. A second group of piglets was treated with dimethylthiourea (DMTU: 750 mg/kg), a putative oxygen radical scavenger. In these animals, GBS increased pulmonary arterial pressure by only 7 ± 1 torr and reduced arterial PO2 by a modest 10 ± 8 torr. Importantly, thromboxane B2 content in venous blood failed to increase above control levels in DMTU-treated animals. The protective effects of DMTU in GBS-treated piglets could not be ascribed to inhibition of cyclooxygenase or thromboxane synthase because the oxygen radical scavenger failed to attenuate increases in pulmonary arterial pressure and venous thromboxane B2 content or reductions in arterial PO2 caused by i.v. infusions of arachidonic acid. DMTU also did not ameliorate pulmonary hypertension evoked by the thromboxane mimetic U44069, thereby suggesting that the scavenger did not act as an end-organ antagonist of thromboxane receptors. These observations suggest that GBS promotes accumulation of thromboxane A2 and attendant pulmonary hypertension through an oxygen radical-dependent mechanism.

Neurosonographic diagnosis of thalamic/basal ganglia vasculopathy in trisomy 13 - An important diagnostic aid

We performed a retrospective review of all the infants diagnosed with trisomy 13 in our institution from 1982 to 1995 and evaluated the neurosonographic findings along with their clinical information and cytogenetic analysis. Nine babies were admitted with trisomy 13. Sonography of the head was performed on 4 patients, and demonstrated in all of them a linear, branching, echogenic pattern in the thalamus/basal ganglia. Doppler evaluation of the thalamus/basal ganglia was performed in 3 of the 4 cases and confirmed these linear echogenicities to be of vascular origin. This is the first study to evaluated the occurrence of this finding in a specific syndrome, namely trisomy 13.

Evidence for hydroxyl radical involvement in group B streptococcus-induced pulmonary hypertension and arterial hypoxemia in young piglets

ABSTRACT: Early onset neonatal GBS infection is associated with pulmonary hypertension, pulmonary edema, and arterial hypoxemia. Although the mechanisms underlying these cardiopulmonary disturbances are not completely understood, multiple lines of evidence suggest that inflammatory mediators may be involved. This study examined the actions of dimethylthiourea (DMTU), a relatively selective scavenger of hydroxyl radical, on GBS-induced pulmonary hypertension, arterial hypoxemia, and pulmonary edema formation in young piglets. Relative to control animals, intravenous infusion of GBS (108 organisms/kg/min for 60 min) provoked sustained increases in pulmonary arterial pressure (Ppa:+88%) and total pulmonary resistance (TPR: 128%). GBS infusion also was associated with profound decreases in arterial Po2 (—58%). Pulmonary edema was present in GBS-treated animals as evidenced by an 8.4% increase in the lung wet-to-dry weight ratio. After pretreatment with DMTU (0.75 g/kg administered intravenously over 30 min), GBS increased Ppa by 33% and TPR by only 16%. Similarly, after DMTU pretreatment GBS decreased arterial oxygen tension by only 12%. DMTU also limited the GBS-induced increase in lung wet-to-dry weight ratio to 2.6% These findings demonstrate that DMTU attenuates GBS-induced pulmonary hypertension, pulmonary edema, and arterial hypoxemia and suggest that hydroxyl radicals play an important role in these cardiopulmonary disturbances.

In Vitro Inactivation of Pulmonary Surfactant Replacement Preparations by Serum Albumin

Inactivation of the surface activity of pulmonary surfactant by serum proteins is an important part of neonatal respiratory distress syndrome. The ability of serum proteins to diminish the surface activity of surfactant preparations used to treat respiratory distress syndrome has not been fully described. The sensitivity of clinically useful pulmonary replacement preparations beractant (Survanta) and colfosceril palmitate, cetyl alcohol, and tyloxapol (Exosurf) to albumin inactivation was examined in vitro by the Wilhelmy plate technique. At a final lipid concentration of 0.1 mg/mL and in the absence of albumin, both Survanta and Exosurf exhibited equilibrium surface tensions in the range of 35 dynes/cm. In the presence of albumin, the surface tension of Survanta was markedly higher. Maximal response of Survanta to albumin was observed at about 1 mg/mL protein concentration. When the lipid concentration was raised to 0.3 mg/mL, the presence of albumin had little effect. With Exosurf, the presence of albumin resulted in only minor elevations of surface tension, even at an albumin concentration 10-fold greater than that used in the experiments with Survanta. These results indicate that at lipid concentrations of 0.1 mg/mL and less, the surface activity of the bovine purified lung surfactant Survanta is more sensitive to the presence of albumin than is the synthetic preparation Exosurf.

Impact of Prostaglandin and Thromboxane Synthesis Blockade on Disposition of Group B Streptococcus in Lung and Liver of Intact Piglet

ABSTRACT: Group B streptococci (GBS) localizing in the lungs of infant piglets is killed in part by an oxygen radical-dependent mechanism (Bowdy BD, Marple SL, Pauly TH, Coonrod JD, Gillespie MN: Am Rev Respir Dis 141:648–653, 1990). The source of bactericidal oxygen radicals is unknown, but cyclooxygenation of arachidonic acid, an initial event in prostanoid synthesis, is accompanied by substantial oxygen radical generation. Because blockade of prostaglandin H synthase (cyclooxygenase) with indomethacin prevents GBS-induced pulmonary hypertension, we reasoned that the salutary effect of indomethacin might be associated with a reduction in the efficacy of bactericidal activity directed against GBS. To address this possibility, the distribution and viability of 111In-labeled GBS (108 colony forming units/kg/min i.v. for 15 min) were assessed in lungs and livers of control piglets, piglets treated with indomethacin (1 mg/kg), and piglets treated with OKY-046 (10 mg/kg), an inhibitor of thromboxane synthase that also forestalls GBS-induced pulmonary hypertension. Relative to control animals, indomethacin treatment increased pulmonary GBS uptake with no change in bacterial distribution into the liver. OKY-046 failed to influence pulmonary bacterial uptake but promoted a substantial increase in GBS depositing in the liver. In contrast to its effects on pulmonary bacterial deposition, indomethacin failed to increase lung bacterial viability relative to control animals. Indomethacin also was without effect on hepatic bacterial viability. OKY-046 failed to influence pulmonary bacterial viability but markedly augmented hepatic GBS viability to the extent that significant bacterial proliferation occurred. Both indomethacin and OKY-046 abolished GBS-induced pulmonary hypertension by preventing the increase in pulmonary arterial pressure and decrease in cardiac output. Viewed collectively, these observations suggest that oxygen radicals derived from cyclooxygenation of arachidonic acid in GBS-treated piglets do not contribute to microbicidal activity directed against the organism.

Bilirubin as an Antioxidant: Effect on Group B Streptococci-Induced Pulmonary Hypertension in Infant Piglets

Bilirubin scavenges toxic oxygen radicals in vitro, but it is not known whether this potential salutary effect can be extended to the intact animal. Accordingly, the present experiments tested the hypothesis that bilirubin protects against oxygen radical-dependent pulmonary hypertension and arterial hypoxemia in piglets infected with group B streptococci (GBS). Piglets ranging in age and weight from 7 to 14 days and 1.5 to 2.0 kg, respectively, were infused for 60 min with 108 cfu GBS/kg/min. One group of 7 animals was pretreated with a bolus infusion of 15 mg/kg of bilirubin followed by a continuous bilirubin infusion. A second group of 7 animals was given the vehicle. While plasma bilirubin levels in control animals were negligible, administration of exogenous bilirubin was associated with plasma levels of 13.0 +/- 0.74 mg%. Piglets treated with exogenous bilirubin exhibited GBS-induced increases in pulmonary arterial pressure and decreases in PaO2 of 16.1 +/- 2.0 and 46.5 +/- 4.3 torr, respectively. In control animals, GBS increased pulmonary arterial pressure and decreased PaO2 by 17.5 +/- 1.6 and 47.9 +/- 3.2 torr, respectively. Neither the peak changes in pulmonary arterial pressure or PaO2 nor the time courses of these alterations differed between treatment groups. These observations indicate that bilirubin fails to prevent GBS-induced pulmonary hypertension and arterial hypoxemia in infant piglets and suggests that in this particular model bilirubin does not exhibit appreciable oxygen radical scavenging activity.

Dimethylthiourea Reverses Sepsis-Induced Pulmonary Hypertension in Piglets

Dimethylthiourea (DMTU), a putative hydroxyl radical scavenger, attenuates thromboxane generation and pulmonary hypertension in the piglet model of group B streptococcal (GBS) sepsis. This study tested the hypothesis that DMTU reverses ongoing GBS-induced pulmonary hypertension coincident with decreased thromboxane production. Piglets (n = 15) received a 60 min infusion of GBS (10(-8) cfu/kg/min). Mean pulmonary artery pressure (Ppa), arterial blood gases (ABGs), and thromboxane B2 (TXB) levels were measured at 10 min intervals throughout the study. GBS infusion resulted in a marked increase in pulmonary artery pressure (mean delta Ppa = 31 mm Hg) and a significant decline in PaO2 (mean = -80 torr) within 10 min of beginning the infusion. pH decreased from a mean of 7.47 to 7.37. DMTU, 750 mg/kg, or normal saline vehicle was infused over 10-15 min beginning 10 min after initiating GBS. Ppa decreased significantly within 10 min of DMTU infusion. Piglets receiving vehicle had a slow decline in Ppa. Piglets receiving DMTU also had an improvement in PaO2 and showed no further drop in pH. Piglets receiving vehicle had no improvement in PaO2 and demonstrated a continued decline in pH. TXB levels did not differ between the groups at any time interval. We conclude that DMTU can partially reverse GBS-induced pulmonary hypertension, but may function through mechanisms independent of thromboxane generation.

Intrinsic Microbicidal Activity and Pulmonary Hypertension in Isolated Newborn Piglet Lungs

ABSTRACT: The lung appears to be one of the dominant sites of bacterial clearance from the blood of infant piglets. Part of the lung bacterial clearance involves activation of an oxygen radical bactericidal mechanism that may be central to induction of acute pulmonary hypertension. The present study determined whether this bactericidal activity was intrinsic to resident lung cells. Isolated piglet lung preparations perfused with blood-free salt solution were used to delineate the amount of group B streptococci (GBS) extracted and killed upon transit through pulmonary vasculature. Approximately 45% of infused GBS was deposited in the lung during a single pulmonary transit, whereas nearly 40% of the organisms sequestered in the lung were killed within a 30-min period. Pretreatment with dimethylthiourea, a scavenger of hydroxyl radical that inhibits GBS-induced pulmonary hypertension, attenuated both bacterial uptake and killing to similar extents. Along with its deposition in the lung, GBS also induced concentration-dependent increases in total pulmonary resistance, which were related principally to increases in upstream arterial resistance. Lung weight also increased in a concentration-dependent manner. Both the increase in total pulmonary resistance and lung weight were temporally related to elevation in perfusion medium content of the stable thromboxane degradation product, thromboxane B2. Pretreatment with indomethacin, a prostaglandin H synthase inhibitor, or sodium(E)-3[4-(l-imidazolyl-methyl)phenyl]-2-propenoic acid a thromboxane synthase inhibitor, reduced GBS-induced pulmonary hypertension and edema. Infusion of thromboxane mimetic, 9,11-dideoxy-9α,11α-epoxymethano prostaglandin F2α, mimicked the ability of GBS to increase total pulmonary and upstream arterial resistances but not development of pulmonary edema. These results suggest that, in isolated piglet lungs, GBS evokes an intrinsic bactericidal response residing within lung cells, probably pulmonary intravascular macrophages, which may be responsible for the initiation of pulmonary hemodynamic changes. It also appears that oxygen radicals play a major role in the bactericidal mechanism directed against GBS. In addition, these results suggest that thromboxane may be the major arachidonic acid metabolite responsible for GBS-induced pulmonary hypertension but perhaps not for development of pulmonary edema.