bluuebunny/crossref_metadata_2025 huggingface dataset:
Spline surface interpolation for calculating 3-D ventricular strains from MRI tissue tagging
Authors: Moulton M. J. , Creswell L. L. , Downing S. W. , Actis R. L. , Szabo B. A. , Vannier M. W. , Pasque M. K.
Published: December 2017

A method is developed and validated for approximating continuous smooth distributions of finite strains in the ventricles from the deformations of magnetic resonance imaging (MRI) tissue tagging "tag lines" or "tag surfaces." Tag lines and intersections of orthogonal tag lines are determined using a semiautomated algorithm. Three-dimensional (3-D) reconstruction of the displacement field on tag surfaces is performed using two orthogonal sets of MRI images and employing spline surface interpolation. The 3-D regional ventricular wall strains are computed from an initial reference image to a deformed image in diastole or systole by defining a mapping or transformation of space between the two states. The resultant mapping is termed the measurement analysis solution and is defined by determining a set of coefficients for the approximating functions that best fit the measured tag surface displacements. Validation of the method is performed by simulating tag line or surface deformations with a finite element (FE) elasticity solution of the heart and incorporating the measured root-mean-square (rms) errors of tag line detection into the simulations. The FE-computed strains are compared with strains calculated by the proposed procedure. The average difference between two-dimensional (2-D) FE-computed strains and strains calculated by the measurement analysis was 0.022 +/- 0.009 or 14.2 +/- 3.6% of the average FE elasticity strain solution. The 3-D displacement reconstruction errors averaged 0.087 +/- 0.002 mm or 2.4 +/- 0.1% of the average FE solution, and 3-D strain fitting errors averaged 0.024 +/- 0.011 or 15.9 +/- 2.8% of the average 3-D FE elasticity solution. When the rms errors in tag line detection were included in the 2-D simulations, the agreement between FE solution and fitted solution was 24.7% for the 2-D simulations and 19.2% for the 3-D simulations. We conclude that the 3-D displacements of MRI tag lines may be reconstructed accurately; however, the strain solution magnifies the small errors in locating tag lines and reconstructing 3-D displacements.
THE DETERMINATION OF THE SURFACE AREA OF WOMEN AND ITS USE IN EXPRESSING BASAL METABOLIC RATE
Authors: Bradfield Hannah Stillman
Published: December 2017

Article by Hannah Stillman Bradfield. By an oversight (p. 570) direct reference was not given to Brody's contribution to the subject. This reference is: Brody and Elting, Research Bulletin 89, Agricultural Experiment Station, University of Missouri, 1926.
FUSILLADE PATTERNS OF INSPIRATORY AND EXPIRATORY MUSCLES AND THEIR EFFECTS ON THE RESPIRATORY ACT
Authors: Gesell Robert
Published: December 2017

Volume 116, No. 1

On page 231, Fig. 7 for Ext. Abd. Obl. read Internal Intercostal.
Cardiac responses to stimulation of thoracic afferents in the primate and canine
Authors: Kostreva DR , Hess GL , Zuperku EJ , Neumark J , Coon RL , Kampine JP
Published: December 2017

Excitatory cardiovascular responses to electrically stimulated upper thoracic sympathetic afferent nerves were observed in halothane-anesthetized mongrel dogs and monkeys. The central end of the transected ventral limb of the left ansa subclavia was stimulated before and after several types of denervation. Significant increases in right and left ventricular maximum systolic pressures, systolic and diastolic systemic blood pressures, and aortic flow were observed. The carotid sinuses were denervated bilaterally and stimulation of the ansa was repeated. The cardiovascular responses to stimulation of the ventral ansa after carotid sinus denervation were greater in magnitude than those observed prior to denervation. This carotid sinus modulation of cardiovascular responses was observed in dogs and monkeys. Cardiovascular responses to stimulation of the ventral ansa after bilateral vagotomy were significantly less than the responses observed after carotid sinus denervation prior to vagotomy. However, the responses after vagotomy were statistically identical to responses obtained while stimulating the ventral ansa when the carotid sinuses and vagi remained intact.
Mechanical properties of rat cardiac muscle during experimental thiamine deficiency
Authors: Cohen EM , Abelmann WH , Messer JV , Bing HL
Published: December 2017

The intrinsic contractile properties of isolated left ventricular trabecular periods of 24, 40, and 53 days were compared with those from 35 weight-matched food-deprived animals and 25 ad libitum fed controls. Contraction mechanics were measured for each muscle when stretched to the peak of its length tension curve at 28 degrees C in oxygenated Krebs-Henseleit solution. Although both thiamine-deficient and food-deprived groups had a marked diminution in body and left ventricular weight in comparison to ad libitum fed controls, no differences in left ventricular weights from thiamine-deficient and food-deprived rats were observed. In comparison to ad libitum fed control animals, food-deprived rats exhibited a prolongation in the duration of muscle contraction and an augmentation of tension development after 40 days. Upon appearance of neurological signs at an average of 53 days of a thiamine-deficient rats exhibited a decrease in performance associated with a decrease in the duration of contraction and rate of tension development.
Dependence of Cr-EDTA absorption from the rumen on luminal osmotic pressure
Authors: Dobson A , Sellers AF , Gatewood VH
Published: December 2017

A method for the measurement of [51Cr]EDTA absorption from the ventral sac of the rumen with an error of the order of +/-10% is described. When a solution present in the rumen was hypotonic or isotonic, the absorption rate of [51Cr]EDTA expressed as a clearance was about 0.2 ml/min. This gave rise to negligible errors when [51Cr]EDTA was used as an unabsorbed marker to calculate net water movements. When the osmotic pressure in the rumen exceeded that of plasma by 30-40 mos-mol/kg, the absorption rate of [51Cr]EDTA appeared to be related to the degree of hypertonicity. Absorption rates as high as 8 ml/min were observed within a range of osmotic pressures normally encountered postprandially in the rumen. Under hypertonic conditions, a correction for the absorption of this large anion was necessary if passage of water into the lumen were not to be systematically overestimated.
Relationships between resting tension and mechanical properties of papillary muscle
Authors: Templeton G , Adcock R , Willerson JT , Nardizzi L , Wildenthal K , Mitchell JH
Published: December 2017

The influence on mechanical properties of changes in resting tension over a range from 0.5 to 4.5 g was investigated in 12 isolated cat papillary muscles. At each resting tension, the muscles contracted isometrically with the exception of an externally applied sinusoidal stretch of 0.5% Lmax (deltaL) and 20 Hz. Stiffness (deltaT/deltaL) was determined from deltaL and the peak amplitudes (deltaT) of the individual cycles from the sinusoidal component of tension. Assuming that the muscle and experimental apparatus behaved as a linear second-order mechanical system, it was possible to divide stiffness into its elastic and viscous components. During rest, total stiffness and its components were linearly related to tension. During contraction, stiffness and its elastic component were linearly related to tension. Furthermore, increasing resting tension increased the intercept and decreased the slope of this linear stiffness-tension relationship. The relationship between viscous stiffness and tension during contraction is more complex in that it is a direct relationship at low resting tensions, but an inverse one at high resting tensions.
Sustained pressore responsiveness to prolonged hypothalamic stimulation in awake rats
Authors: Bunag RD , Riley E , Montello M
Published: December 2017

Whether or not pressor responsiveness changes in unanesthetized rats during recurrent sympathetic excitation was determined by recording blood pressure and heart rate continuously while the posterior hypothalamus was stimulated repeatedly with constant currents. Because preliminary tests showed that telestimulation with a radio-controlled stimulator produced erratic responses, awake rats were routinely stimulated in a conventional manner by connecting them through wires to a square-wave stimulator. Although tachycardia was the most common chronotropic effect, bradycardia also occurred, and both responses were occasionally seen in the same rat at different times. Inhibition of chronotropic responses by combined pharmacologic blockage with propranolol and atropine did not affect corresponding pressure responses in normotensive rats. Renal and spontaneously hypertensive rats always had larger pressor responses than normotensive ones, and, in spite of individual variations, responsiveness generally remained unaltered during 3-6 h of repeated hypothalamic stimulation. These results indicate that in awake normotensive or hypertensive rats cardiovascular responses to posterior hypothalamic stimulation continue unabated even when stimulation is repeated for hours.
A role for glucose in hypothermic hamsters
Authors: Resch GE , Musacchia XJ
Published: December 2017

Hamsters undergo hypothermia when exposed to a mixture of 80% helium and 20% oxygen at low ambient temperatures. The hypothermic hamster, rectal temperature (Tre) 7 degrees C, becomes hypoglycemic, and reversal of hypoglycemia is effected with glucose infusion. Hypothermic hamsters at Tre 7 degrees C showed a fivefold increase in survival times from 20 to 100.5 h when infused with glucose which maintained a blood level at about 45 mg/100 ml. A potential role for osmotic effects of the infusion was tested and eliminated. There was no improvement in survival of 3-O-methylglucose or dextran 40-infused animals. The fact that death eventually occurs even in the glucose-infused animal after about 4 days and that VO2 undergoes a slow decrement in that period suggests that hypothermic survival is not wholly substrate limited. Radioactive tracer, [U-14C]glucose, showed that localization of the 14C, was greatest in brain tissue and diaphragm, intermediate in heart and kidney, and lowest in skeletal muscle and liver. The significance of the label at sites important to respiration and circulation was presented.
Effect of hypermagnesemia on rat jejunal sodium and water transport
Authors: DiBona GF , Sawin LL
Published: December 2017

Hypermagnesemia decreases sodium and water reabsorption in the rat renal proximal tubule. To further understand this action, the effect of hypermagnesemia on sodium and water transport in the in vivo perfused rat jejunum was studied. The rat jejunum was chosen as another transporting epithelial tissue in the same species with unidirectional sodium flux characteristics similar to the rat renal proximal tubule, i.e., leaky as opposed to tight. Hypermagnesemia decreased net jejunal sodium and water reabsorption. This decrease was due to a reduction in unidirectional sodium efflux from lumen to blood and not to an increase in unidirectional sodium influx from blood to lumen. Hypermagnesemia did not change the jejunal permeability to inulin. The effect of hypermagnesemia on jejunal sodium and water transport is similar to that renal proximal tubule sodium and water transport. This similarity suggests that the mechanism of action of magnesium of these two transporting epithelial tissues is similar.
Dietary self-selection following subdiaphragmatic vagotomy in the white rat
Authors: Fox KA , Kipp SC , VanderWeele DA
Published: December 2017

Nineteen female Wistar rats were employed in a self-selection-of-diet study assessing the effects of vagotomy on macronutrient consumption. Nutrients offered were a 30% sucrose solution, a 15% casein hydrolysate solution, and olive oil. In addition, noncaloric bulk, vitamins, and water were allowed. Bilaterally, subdiaphragmatically vagotomized rats showed a significant decrement in carbohydrate ingestion and daily total caloric intake (P less than .05 in both comparisons). Fat and protein intakes were unreliably altered. It is hypothesized that the vagus nerve relays glucose absorption and/or storage information from the periphery to the brain (perhaps lateral hypothalamusy, and hence, vagotomy produces carbohydrate-intake decrements, specifically. The decrement could also be explained as resulting from efferent effects of vagotomy such as decreased gut hormonal release.
Mechanism of pellet egestion in great-horned owls (Bubo virginianus)
Authors: Duke GE , Evanson OA , Redig PT , Rhoades DD
Published: December 2017

To study the mechanism of oral pellet egestion in great-horned owls, bipolar electrodes and strain-gauge transducers were chronically implanted in the esophagus, muscular stomach, and duodenum of six owls. Recordings from conscious owls plus simultaneous radiographic observations revealed characteristic gastrointestinal motility patterns associated with egestion. Beginning at about 12 min before egestion, gastric contractions formed the final shape of the pellet and pushed it into the lower esophagus. The pellet was moved out of the esophagus by antiperistalsis during the last 8--10 s before egestion. During pellet egestion, contractions of abdominal muscles were not detected. Pellet egestion appears to be unlike either emesis in mammals with a simple stomach or regurgitation in ruminants.
Coronary intercapillary distance during growth: relation to PtO2 and aerobic capacity
Authors: Henquell L , Odoroff CL , Honig CR
Published: December 2017

Intercapillary distance (ICD) was measured in left ventricles of rats beating in situ. Between 40 and 400 days of age, left ventricular weight increased threefold and ICD increased from 12.5-19.5 mum. ICD could be decreased by at least 2 mum at all ages studied. The number of capillaries which must be recruited to reduce ICD by 2 mum fell from 1,200/mm2 at 40 days to 280/mm2 at 400 days. Ventricular growth did not affect the O2 sensitivity of precapillary sphincters or the uniformity of capillary spacing. Calculations indicate that under basal conditions tissue PO2 (Pto2) in subepicardium is about the same at 40 and 400 days, even though VO2 per gram, capillary density, and ICD change twofold, twofold, and 7 mum, respectively. Nevertheless, as the ventricle grows, capillary recruitment becomes progressively less effective in defending Pto2 under conditions of stress. Diminished coronary capillary compensation for stress may, in part, account for the effect of age on the maximum aerobic capacity of the whole animal.
Parallel secretion of digestive enzymes by the in vitro rabbit pancreas
Authors: Steer ML , Glazer G
Published: December 2017

Nonparallel discharge of digestive enzymes by the in vitro rabbit pancreas has been previously reported and this observation has been used to challenge the mass transport theory of enzyme synthesis and secretion by the exocrine pancreas. The current investigation reexamines the pattern of enzyme secretion from the in vitro rabbit pancreas. Secretion of alpha-amylase, trypsinogen, and chymotrypsinogen was studied from the unstimulated gland and from glands stimulated with either methacholine, cholecystokinin-pancreozymin, or the active octapeptide of cholecystokinin-pancreozymin. In all cases, only parallel discharge of alpha-amlyse, trypsinogen, and chymotrypsinogen was noted. These observations are consistent with the mass-transport theory.
Role of neuronal nitric oxide synthase in regulation of vascular and ductus arteriosus tone in the ovine fetus
Authors: Rairigh Robyn L. , Storme Laurent , Parker Thomas A. , le Cras Timothy D. , Markham Neil , Jakkula Malathi , Abman Steven H.
Published: December 2017

Nitric oxide (NO) is produced by NO synthase (NOS) and contributes to the regulation of vascular tone in the perinatal lung. Although the neuronal or type I NOS (NOS I) isoform has been identified in the fetal lung, it is not known whether NO produced by the NOS I isoform plays a role in fetal pulmonary vasoregulation. To study the potential contribution of NOS I in the regulation of basal fetal pulmonary vascular resistance (PVR), we studied the hemodynamic effects of a selective NOS I antagonist, 7-nitroindazole (7-NINA), and a nonselective NOS antagonist, N-nitro-l-arginine (l-NNA), in chronically prepared fetal lambs (mean age 128 ± 3 days, term 147 days). Brief intrapulmonary infusions of 7-NINA (1 mg) increased basal PVR by 37% ( P < 0.05). The maximum increase in PVR occurred within 20 min after infusion, and PVR remained elevated for up to 60 min. Treatment with 7-NINA also increased the pressure gradient between the pulmonary artery and aorta, suggesting constriction of the ductus arteriosus (DA). To test whether 7-NINA treatment selectively inhibits the NOS I isoform, we studied the effects of 7-NINA andl-NNA on acetylcholine-induced pulmonary vasodilation. The vasodilator response to acetylcholine remained intact after treatment with 7-NINA but was completely inhibited after l-NNA, suggesting minimal effects on endothelial or type III NOS after 7-NINA infusion. Western blot analysis detected NOS I protein in the fetal lung and great vessels including the DA. NOS I protein was detected in intact and endothelium-denuded vessels, suggesting that NOS I is present in the medial or adventitial layer. We conclude that 7-NINA, a selective NOS I antagonist, increases basal PVR, systemic arterial pressure, and DA tone in the late-gestation fetus and that NOS I protein is present in the fetal lung and great vessels. We speculate that NOS I may contribute to NO production in the regulation of basal vascular tone in the pulmonary and systemic circulations and the DA.
Cell growth modulates nitric oxide synthase expression in fetal pulmonary artery endothelial cells
Authors: Whitney Jeannette A. , German Zohre , Sherman Todd S. , Yuhanna Ivan S. , Shaul Philip W.
Published: December 2017

Nitric oxide (NO), produced by endothelial (e) nitric oxide synthase (NOS), is a critical mediator of vascular function and growth in the developing lung. Pulmonary eNOS expression is diminished in conditions associated with altered pulmonary vascular development, suggesting that eNOS may be modulated by changes in pulmonary artery endothelial cell (PAEC) growth. We determined the effects of cell growth on eNOS expression in cultured ovine fetal PAEC studied at varying levels of confluence. NOS enzymatic activity was sixfold greater in quiescent PAEC at 100% confluence compared with more rapidly replicating cells at 50% confluence. To determine if there is a reciprocal effect of NO on PAEC growth, studies of NOS inhibition or the provision of exogenous NO from spermine NONOate were performed. Neither intervention had a discernable effect on PAEC growth. The influence of cell growth on NOS activity was unique to pulmonary endothelium, because varying confluence did not alter NOS activity in fetal systemic endothelial cells. The effects of cell growth induced by serum stimulation were also evaluated, and NOS enzymatic activity was threefold greater in quiescent, serum-deprived cells compared with that in serum-stimulated cells. The increase in NOS activity observed at full confluence was accompanied by parallel increases in eNOS protein and mRNA expression. These findings indicate that eNOS gene expression in fetal PAEC is upregulated during cell quiescence and downregulated during rapid cell growth. Furthermore, the interaction between cell growth and NO in the PAEC is unidirectional.
Cl-channel activation is necessary for stimulation of Na transport in adult alveolar epithelial cells
Authors: O'Grady Scott M. , Jiang Xinpo , Ingbar David H.
Published: December 2017

In this review, we discuss evidence that supports the hypothesis that adrenergic stimulation of transepithelial Na absorption across the alveolar epithelium occurs indirectly by activation of apical Cl channels, resulting in hyperpolarization and an increased driving force for Na uptake through amiloride-sensitive Na channels. This hypothesis differs from the prevailing idea that adrenergic-receptor activation increases the open probability of Na channels, leading to an increase in apical membrane Na permeability and an increase in Na and fluid uptake from the alveolar space. We review results from cultured alveolar epithelial cell monolayer experiments that show increases in apical membrane Cl conductance in the absence of any change in Na conductance after stimulation by selective β-adrenergic-receptor agonists. We also discuss possible reasons for differences in Na-channel regulation in cells grown in monolayer culture compared with that in dissociated alveolar epithelial cells. Finally, we describe some preliminary in vivo data that suggest a role for Cl-channel activation in the process of amiloride-sensitive alveolar fluid absorption.
High K+-induced membrane depolarization attenuates endothelium-dependent pulmonary vasodilation
Authors: Seiden Jan E. , Platoshyn Oleksandr , Bakst Alan E. , McDaniel Sharon S. , Yuan Jason Xiao-Jian
Published: December 2017

Impairment of endothelium-dependent pulmonary vasodilation has been implicated in the development of pulmonary hypertension. Pulmonary vascular smooth muscle cells and endothelial cells communicate electrically through gap junctions; thus, membrane depolarization in smooth muscle cells would depolarize endothelial cells. In this study, we examined the effect of prolonged membrane depolarization induced by high K+ on the endothelium-dependent pulmonary vasodilation. Isometric tension was measured in isolated pulmonary arteries (PA) from Sprague-Dawley rats, and membrane potential was measured in single PA smooth muscle cells. Increase in extracellular K+ concentration from 4.7 to 25 mM significantly depolarized PA smooth muscle cells. The 25 mM K+-mediated depolarization was characterized by an initial transient depolarization (5–15 s) followed by a sustained depolarization that could last for up to 3 h. In endothelium-intact PA rings, ACh (2 μM), levcromakalim (10 μM), and nitroprusside (10 μM) reversibly inhibited the 25 mM K+-mediated contraction. Functional removal of endothelium abolished the ACh-mediated relaxation but had no effect on the levcromakalim- or the nitroprusside-mediated pulmonary vasodilation. Prolonged (∼3 h) membrane depolarization by 25 mM K+ significantly inhibited the ACh-mediated PA relaxation (−55 ± 4 vs. −29 ± 2%, P < 0.001), negligibly affected the levcromakalim-mediated pulmonary vasodilation (−92 ± 4 vs. −95 ± 5%), and slightly but significantly increased the nitroprusside-mediated PA relaxation (−80 ± 2 vs. 90 ± 3%, P < 0.05). These data indicate that membrane depolarization by prolonged exposure to high K+ concentration selectively inhibited endothelium-dependent pulmonary vasodilation, suggesting that membrane depolarization plays a role in the impairment of pulmonary endothelial function in pulmonary hypertension.
Phospholipid molecular species of bronchoalveolar lavage fluid after local allergen challenge in asthma
Authors: Heeley Emma L. , Hohlfeld Jens M. , Krug Norbert , Postle Anthony D.
Published: December 2017

Electrospray ionization mass spectrometry was used to quantify phosphatidylcholine (PC) and phosphatidylglycerol (PG) molecular species in bronchoalveolar lavage fluid (BALF) from control and mild asthmatic subjects after local allergen challenge. BALF was obtained from 5 control and 13 asthmatic subjects before and 24 h after segmental allergen and saline challenge. There were no differences in the ratio of total PC to total PG or in the molecular species composition of PC or PG between the asthmatic and control groups under basal conditions. Allergen challenge in asthmatic but not in control volunteers caused a significant increase in the PC-to-PG ratio because of increased concentrations of PC species containing linoleic acid (16:0/18:2 PC, 18:0/18:2 PC, and 18:1/18:2 PC). These molecular species were characteristic of plasma PC analyzed from the same subjects, strongly suggesting that the altered PC composition in BALF in asthmatic subjects after allergen challenge was due to infiltration of plasma lipoprotein, not to catabolism of surfactant phospholipid. Interactions between surfactant and lipoprotein infiltrate may contribute to surfactant dysfunction and potentiate disease severity in asthma.
Estradiol improves pulmonary hemodynamics and vascular remodeling in perinatal pulmonary hypertension
Authors: Parker Thomas A. , Dunbar Ivy D. , Galan Henry L. , Grover Theresa R. , Kinsella John P. , Abman Steven H.
Published: December 2017

Partial ligation of the ductus arteriosus (DA) in the fetal lamb causes sustained elevation of pulmonary vascular resistance (PVR) and hypertensive structural changes in small pulmonary arteries, providing an animal model for persistent pulmonary hypertension of the newborn. Based on its vasodilator and antimitogenic properties in other experimental studies, we hypothesized that estradiol (E2) would attenuate the pulmonary vascular structural and hemodynamic changes caused by pulmonary hypertension in utero. To test our hypothesis, we treated chronically instrumented fetal lambs (128 days, term = 147 days) with daily infusions of E2 (10 μg; E2 group, n = 6) or saline (control group, n = 5) after partial ligation of the DA. We measured intrauterine pulmonary and systemic artery pressures in both groups throughout the study period. After 8 days, we delivered the study animals by cesarean section to measure their hemodynamic responses to birth-related stimuli. Although pulmonary and systemic arterial pressures were not different in utero, fetal PVR immediately before ventilation was reduced in the E2-treated group (2.43 ± 0.79 vs. 1.48 ± 0.26 mmHg ⋅ ml−1 ⋅ min, control vs. E2, P < 0.05). During the subsequent delivery study, PVR was lower in the E2-treated group in response to ventilation with hypoxic gas but was not different between groups with ventilation with 100% O2. During mechanical ventilation after delivery, arterial partial O2 pressure was higher in E2 animals than controls (41 ± 11 vs. 80 ± 35 Torr, control vs. E2, P < 0.05). Morphometric studies of hypertensive vascular changes revealed that E2 treatment decreased wall thickness of small pulmonary arteries (59 ± 1 vs. 48 ± 1%, control vs. E2, P < 0.01). We conclude that chronic E2 treatment in utero attenuates the pulmonary hemodynamic and histological changes caused by DA ligation in fetal lambs.
Hepatocyte growth factor is elevated in chronic lung injury and inhibits surfactant metabolism
Authors: Vivekananda Jeevalatha , Awasthi Vibhudutta , Awasthi Shanjana , Smith Dolphin B. , King Richard J.
Published: December 2017

Adult respiratory distress syndrome may incorporate in its pathogenesis the hyperplastic proliferation of alveolar epithelial type II cells and derangement in synthesis of pulmonary surfactant. Previous studies have demonstrated that hepatocyte growth factor (HGF) in the presence of serum is a potential mitogen for adult type II cells (R. J. Panos, J. S. Rubin, S. A. Aaronson, and R. J. Mason. J. Clin. Invest. 92: 969–977, 1993) and that it is produced by fetal mesenchymal lung cells (J. S. Rubin, A. M.-L. Chan, D. P. Botarro, W. H. Burgess, W. G. Taylor, A. C. Cech, D. W. Hirschfield, J. Wong, T. Miki, P. W. Finch, and S. A. Aaronson. Proc. Natl. Acad. Sci. USA 88: 415–419, 1991). In these studies, we expand on this possible involvement of HGF in chronic lung injury by showing the following. First, normal adult lung fibroblasts transcribe only small amounts of HGF mRNA, but the steady-state levels of this message rise substantially in lung fibroblasts obtained from animals exposed to oxidative stress. Second, inflammatory cytokines produced early in the injury stimulate the transcription of HGF in isolated fibroblasts, providing a plausible mechanism for the increased amounts of HGF seen in vivo. Third, HGF is capable of significantly inhibiting the synthesis and secretion of the phosphatidylcholines of pulmonary surfactant. Fourth, HGF inhibits the rate-limiting enzyme in de novo phosphatidylcholine synthesis, CTP:choline-phosphate cytidylyltransferase (EC 2.7.7.15 ). Our data indicate that fibroblast-derived HGF could be partially responsible for the changes in surfactant dysfunction seen in adult respiratory distress syndrome, including the decreases seen in surfactant phosphatidylcholines.
Hypoxia induces hexokinase II gene expression in human lung cell line A549
Authors: Riddle Suzette R. , Ahmad Aftab , Ahmad Shama , Deeb Samir S. , Malkki Mari , Schneider B. Kelly , Allen Corrie B. , White Carl W.
Published: December 2017

During adaptation to hypoxic and hyperoxic conditions, the genes involved in glucose metabolism are upregulated. To probe involvement of the transcription factor hypoxia-induced factor-1 (HIF-1) in hexokinase (HK) II expression in human pulmonary cells, A549 cells and small-airway epithelial cells (SAECs) were exposed to stimuli such as hypoxia, deferoxamine (DFO), and metal ions. The largest increase in HK-II (20-fold for mRNA and 2.5-fold for enzymatic activity) was observed in A549 cells when exposed to DFO. All stimuli selectively increased the 5.5-kb rather than 4-kb transcript in A549 cells. Cycloheximide and actinomycin D inhibited these responses. In addition, cells were transfected with luciferase reporter constructs driven by the full-length HK-II 5′-regulatory region (4.0 kb) or various deletions of that region. A549 cells transfected with the 4.0-kb construct and exposed to hypoxia or DFO increased their luciferase activity 7- and 10-fold, respectively, indicating that HK-II induction is, at least in part, due to increased gene transcription. Sixty percent of the inducible activity of the 4.0-kb construct was shown to reside within the proximal 0.5 kb. Additionally, cotransfection with a stable HIF-1 mutant and the 4.0-kb promoter construct resulted in increased luciferase activity under normoxic conditions. These results strongly suggest that HK-II is selectively regulated in pulmonary cells by a HIF-1-dependent mechanism.
Pulmonary arterial smooth muscle cells modulate cytokine- and LPS-induced cytotoxicity in endothelial cells
Authors: Johnson Bruce A. , Pitt Bruce R. , Davies Paul
Published: December 2017

Cytokines and lipopolysaccharide (LPS) are known to be injurious to vascular endothelial cells (ECs), but the influence of adjacent vascular smooth muscle cells (SMCs) on this injury is unknown. Exposure of cultured rat (RPMECs) or human (HPMECs) pulmonary microvascular ECs on tissue culture plastic to a mixture of cytokines (interleukin-1β, tumor necrosis factor-α, and interferon-γ) and LPS (cytomix) resulted in a significant increase in51Cr release to 35–40%. When unstimulated RPMECs were cocultured with cytomix-pretreated rat pulmonary microvascular SMCs (RPMSMCs) there was an increase in 51Cr release to 8.4%, which was nitric oxide dependent. However, when RPMECs or HPMECs were stimulated in direct contact with their respective SMCs, rather than a further increase in cytomix-induced injury (e.g., >35–40%), 51Cr release decreased to <10%. This cytoprotection was fully reproduced with fixed RPMSMCs, and partially reproduced by plating HPMECs on gelatin. These data show that the direct toxicity of a cytokine and endotoxin mixture on cultured ECs can be reduced by contact with vascular smooth muscle.
Construction and characterization of a chimeric receptor containing the cytoplasmic domain of MUC1 mucin
Authors: Meerzaman Daoud , Xing P. X. , Kim K. Chul
Published: December 2017

MUC1 mucin is a transmembrane glycoprotein that is highly expressed in various cancer cell lines and is also present in most of the glandular epithelial cells including the airway. Although the presence of numerous phosphorylation sites in its cytoplasmic domain suggests its potential role as a receptor, the unavailability of a ligand for MUC1 mucin has limited our understanding of its function. In this paper, we tried to circumvent this problem by constructing a chimeric receptor containing the cytoplasmic domain of MUC1 mucin, which can be phosphorylated on activation. To this end, we constructed a chimeric plasmid vector (pCD8/MUC1) by replacing the extracellular and transmembrane domains of human MUC1 mucin with those of human CD8. Transient transfection of the vector into COS-7 cells resulted in expression of the chimeric receptor on the surface of the COS-7 cells as judged by immunologic assays with various antibodies as well as by fluorescence-activated cell-sorting analysis. Treatment of the transfected COS-7 cells with an anti-CD8 antibody resulted in a significant increase in phosphorylation of tyrosine moieties of the chimeric receptor. This chimeric receptor will serve as a powerful tool in elucidating the signaling mechanism as well as the functional role of MUC1 mucin in the airway.
Angiogenesis and morphogenesis of murine fetal distal lung in an allograft model
Authors: Schwarz Margaret A. , Zhang Fangrong , Lane John E. , Schachtner Susan , Jin Yangsun , Deutsch Gail , Starnes Vaughn , Pitt Bruce R.
Published: December 2017

Neovascularization is crucial to lung morphogenesis; however, factors determining vessel growth and formation are poorly understood. The goal of our study was to develop an allograft model that would include maturation of the distal lung, thereby ultimately allowing us to study alveolar development, including microvascular formation. We transplanted 14-day gestational age embryonic mouse lung primordia subcutaneously into the back of nude mice for 3.5–14 days. Lung morphogenesis and neovascularization were evaluated by light microscopy, in situ hybridization, and immunohistochemical techniques. Embryonic 14-day gestational age control lungs had immature structural features consistent with pseudoglandular stage of lung development. In contrast, 14 days after subcutaneous transplantation of a 14-day gestational age lung, the allograft underwent significant structural morphogenesis and neovascularization. This was demonstrated by continued neovascularization and cellular differentiation, resulting in mature alveoli similar to those noted in the 2-day postnatal neonatal lung. Confirmation of maturation of the allograft was provided by progressive type II epithelial cell differentiation as evidenced by enhanced local expression of mRNA for surfactant protein C and a threefold ( P < 0.008) increase in vessel formation as determined by immunocytochemical detection of platelet endothelial cell adhesion molecule-1 expression. Using the tyrosine kinase Flk-1 receptor ( flk-1) LacZ transgene embryos, we determined that the neovascularization within the allograft was from the committed embryonic lung endothelium. Therefore, we have developed a defined murine allograft model that can be used to study distal lung development, including neovascularization. The model may be useful when used in conjunction with an altered genetic background (knockout or knock in) of the allograft and has the further decided advantage of bypassing placental barriers for introduction of pharmacological agents or DNA directly into the lung itself.
Increased expression of inflammatory mediators in small-airway epithelium from tobacco smokers
Authors: Takizawa Hajime , Tanaka Mitsuru , Takami Kazutaka , Ohtoshi Takayuki , Ito Koji , Satoh Masaru , Okada Yasumasa , Yamasawa Fumihiro , Umeda Akira
Published: December 2017

To study the inflammatory responses of small-airway epithelium in smokers, we harvested enough living epithelial cells (1.97 × 106± 0.74 × 106) with a new ultrathin fiberscope from the very peripheral airways of 22 current smokers and 17 subjects who never smoked after informed consent was obtained. The cells were keratin positive and composed mainly of nonciliated cells. The expression levels of inflammatory markers [interleukin (IL)-8 and intercellular adhesion molecule (ICAM)-1] were evaluated with RT-PCR. The magnitude of the mRNA levels corrected by β-actin transcripts of IL-8 and ICAM-1 was significantly higher in the smokers than in the nonsmokers ( P < 0.001). Furthermore, among current smokers, IL-8 mRNA levels correlated positively with the extent of smoking history [in pack ⋅ years (packs/day × no. of years of smoking); r = 0.754, P < 0.001]. Spontaneously released IL-8 and soluble ICAM-1 levels ( n = 12) from cultured epithelial cells were elevated in subjects with a smoking history than in those without it (IL-8, 1,580 ± 29.6 vs. 354 ± 39.4 pg ⋅ 106cells−1⋅ 24 h−1; P < 0.001; soluble ICAM-1, 356.0 ± 45.9 vs. 112.9 ± 12.9 pg ⋅ 106cells−1⋅ 24 h−1; P < 0.01 by Student's t-test ). In contrast, the epithelial cells from the main bronchi did not show such differences between smokers and nonsmokers. Our study highlighted a close link between smoking and the expression of inflammatory mediators such as IL-8 and ICAM-1 in small airways. Our results also suggested that this new ultrathin bronchofiberscope promised a good approach for the evaluation of cellular changes in the small airways.
Mechanical strain and dexamethasone selectively increase surfactant protein C and tropoelastin gene expression
Authors: Nakamura Tomohiko , Liu Mingyao , Mourgeon Eric , Slutsky Art , Post Martin
Published: December 2017

Physical forces derived from fetal breathing movements and hormones such as glucocorticoids are implicated in regulating fetal lung development. To elucidate whether the different signaling pathways activated by physical and hormonal factors are integrated and coordinated at the cellular and transcriptional levels, organotypic cultures of mixed fetal rat lung cells were subjected to static culture or mechanical strain in the presence and absence of dexamethasone. Tropoelastin and collagen type I were used as marker genes for fibroblasts, whereas surfactant protein (SP) A and SP-C were used as marker genes for distal epithelial cells. Mechanical strain, but not dexamethasone, significantly increased SP-C mRNA expression. Tropoelastin mRNA expression was upregulated by both mechanical strain and dexamethasone. No additive or synergistic effect was observed when cells were subjected to mechanical stretch in the presence of dexamethasone. Neither mechanical strain nor dexamethasone alone or in combination had any significant effect on the expression of SP-A mRNA. Dexamethasone decreased collagen type I mRNA expression, whereas mechanical strain had no effect. The increases in tropoelastin and SP-C mRNA levels induced by mechanical strain and/or dexamethasone were accompanied by increases in their heterogeneous nuclear RNA. In addition, the stretch- and glucocorticoid-induced alterations in tropoelastin and SP-C mRNA expression were abrogated with 10 μg/ml actinomycin D. These findings suggest that tropoelastin and SP-C genes are selectively stimulated by physical and/or hormonal factors at the transcriptional level in fetal lung fibroblasts and distal epithelial cells, respectively.
Distribution of adenine nucleotides in the perfused rat heart
Authors: Kohn M. C. , Achs M. J. , Garfinkel D.
Published: December 2017

A computer technique for determination of the distribution of adenine nucleotides among compartmented, protonated, and metal-chelated species has been developed for the perfused rat heart. This procedure requires knowledge of tissue levels of creatine, creatine phosphate, ATP, ADP, and AMP and the glycolytic and respiration rates. The method is applicable to any physiological state of the organ and has been applied to transient behavior in aerobic, anoxic, and ischemic hearts. The results suggest that ADP uptake and ATP export by mitochondria are normally linked and equal in rate during aerobic metabolism or short-term anoxia but become separate and unequal during ischemia, so that mitochondrial adenine nucleotides, primarily AMP, accumulate.
Blood pressure vulnerability to volume contraction: regulation by adrenal cortical hormones
Authors: Waldinger T. , Seaton J. F. , Harrison T. S.
Published: December 2017

Although the unique sensitivity of blood pressure to hemorrhage after adrenalectomy can be overcome by adrenocortical hormones the specificity of this steroid effect is not known. Under pentobarbital anesthesia five groups of six adult male Sprague-Dawley rats were bled into a 0.9% saline-primed pressure-balanced reservoir. A mean arterial pressure of 50 Torr was maintained for 3 h. When compared to normal rats, those with adrenalectomy showed significantly lowered bleeding volume (P less than 0.05 to less than 0.001). In untreated adrenalectomized rats 120 min following hemorrhage, saline was consistently taken up from the reservoir to maintain blood pressure at 50 Torr. Deoxycorticosterone-cortisol (intravenous bolus coupled with infusion) restored bleeding volume to normal in adrenalectomized rats. Aldosterone (constant infusion) also protected bleeding volume after adrenalectomy, P less than 0.05 to less than 0.01, but this effect was significantly less striking than that of cortisol 180 min after the onset of bleeding, P less than 0.001. Sham adrenalectomy did not affect bleeding volume. Aldosterone appears to be effective in restoring normal bleeding volume sensitivity after adrenalectomy but this effect of aldosterone is less sustained than that seen with cortisol.
Kinetic analysis of calcium desaturation curves from isolated kidney cells
Authors: Uchikawa T. , Borle A. B.
Published: December 2017

This paper provides mathematical solutions for calculating calcium fluxes and compartments in isolated kidney cells from calcium-45 desaturation curves. In contrast to other available methods, these solutions allow the calculation of kinetic parameters even if isotopic equilibrium has not been reached at the beginning of the desaturation period. To test the validity of the method, calcium-45 desaturation experiments were performed in isolated kidney cells after labeling periods of 30, 60, 90, or 120 min. Even though the calcium-45 desaturation curves differ with different labeling times, the respective values of the calculated parameters are practically identical. The optical labeling period was found to lie between 60 and 120 min. Since this new method does not require initial isotopic steady state, it allows a formal kinetic analysis of tracer desaturation curves even after short periods of labeling.
Studies of calcium-45 desaturation from kidney slices in flow-through chambers
Authors: Uchikawa T. , Borle A. B.
Published: December 2017

This paper describes a method to measure calcium fluxes and calcium exchangeable pools in tissue slices by continuous perifusion in flow-through chambers. 45Ca desaturation from rat kidney slices can be analyzed as in an open three-compartment catenary system. A set of equations is given to calculate all the relevant kinetic parameters from the triple exponential equations which best fit the desaturation curves. The results show that the kinetic parameters obtained in kidney slices by this new method are in the same order of magnitude as those previously observed in cultured monkey kidney cells.
Circadian rhythms in blinded rats: correlation between pineal and activity cycles
Authors: Pohl C. R. , Gibbs F. P.
Published: December 2017

The rat pineal gland exhibits diurnal rhythms in levels of N-acetyltransferase activity and its substrate serotonin. We attempted to demonstrate the endogenous nature of these changes by measuring the pineal enzyme and its substrate in rats blinded for 37 and 60 days. In order to determine the proper sampling times for these one-time, terminal measurements, the running activity rhythm of each rat was monitored continuously and the animals were killed at either midrest or midrun. Circadian changes of pineal N-acetyltransferase and serotonin were demonstrated, with enzyme levels high and substrate content low during midrun. Absolute values during each activity phase were similar to those of control rats entrained to a light cycle (LD 12:12). Levels of the pineal constituents were unrelated to local time. These results suggest that rats blinded for up to 60 days maintain their free-running pineal rhythms with undamped amplitudes and in synchronization with the activity rhythm.
Fasting, adrenalectomy, and gluconeogenesis in the chicken and a carnivorous bird
Authors: Veiga J. A. , Roselino E. S. , Migliorini R. H.
Published: December 2017

Previous studies showed that livers from carnivorous birds have a higher gluconeogenic capacity and higher levels of gluconeogenic enzymes than livers from granivorous birds. In this work we compare the effects of fasting and adrenalectomy on gluconeogenesis. Fasting in the chicken elicited increased rates of incorporation of 14C from alanine into blood glucose, increased gluconeogenesis in liver slices, and increased activities of four gluconeogenic enzymes: glucose-6-phosphatase, phosphoenolpyruvate carboxykinase, alanine aminotransferase, and aspartate aminotransferase. These responses in the chicken resemble those observed in fasted rodents. In marked contrast, fasting in black vultures induced decreased rates of incorporation of alanine label into circulating glucose, decreased gluconeogenesis in liver slices, and no change in any of the four enzymes studied. This unusual response to fasting in the carnivorous bird is probably related to the high-protein-low-carbohydrate content of the diet. Fasted adrenalectomized birds (granivorous and carnivorous) had reduced rates of in vivo glucose synthesis, decreased liver gluconeogenesis, and lower activity of glucose-6-phosphatase and aspartate aminotransferase, without change in phosphoenolpyruvate carboxykinase and alanine aminotransferase activities.
Comparison of synchronization of primate circadian rhythms by light and food
Authors: Sulzman F. M. , Fuller C. A. , Moore-Ede M. C.
Published: December 2017

Several circadian rhythms in squirrel monkeys (Saimiri sciureus) entrained by two different agents were studied to compare their mode of coupling with the environmental zeitgebers. Synchronization was accomplished either by light-dark cycles consisting of 12 h of 600 lx followed by 12 h of less than 1 lx (LD 12:12), or by eat-fast cycles in which the animals could eat for 3 h and then had to fast for the remaining 21 h each day (EF 3:21). The rhythms of drinking, colonic temperature, and urinary potassium and water excretion were measured in chair-acclimatized monkeys. The drinking and urinary rhythms were more reproducible (smaller mean variance) and more stable (smaller standard deviation of the timing of a phase reference point) in EF than in LD cycles, whereas the temperature rhythm was more tightly controlled by LD cycles than by EF cycles. In constant light an 8-h phase delay in the EF cycle caused the drinking and urinary rhythms to resynchronize to the EF cycle within one day, while the temperature rhythm required about 6 days to resynchronize. In contrast, previously published data for a similar phase delay in the LD cycle with food available ad libitum show that the drinking and temperature rhythms resynchronized more rapidly than the urinary rhythms. These results indicate that separate mechanisms are involved in transducing temporal cues from LD and EF cycles in the circadian timekeeping system of these nonhuman primates.
Controlling the highest lactate dehydrogenase activity known in nature
Authors: Guppy M. , Hochachka P. W.
Published: December 2017

In the shipjack, Euthynnus pelamis, white muscle appears to possess a powerful anaerobic capacity as well as a significant carbohydrate based aerobic potential. Lactate dehydrogenase occurs at higher activities than found thus far anywhere else in nature and clearly functions in anaerobic glycolysis. Alpha-glycerophosphate dehydrogenase also occurs in unusually high activities and appears to play a role in aerobic glycolysis. Regulation of these two reactions is accomplished by temperature, pH, and creatine phosphate levels. High temperature, low pH, and low creatine phosphate levels all appear to favor lactate dehydrogenase over alpha-glycerophosphate dehydrogenase; low temperature, high pH, and high creatine-phosphate levels, all expected during the quiescent state in this species, and when metabolism in aerobic, all favor alpha-glycerophosphate dehydrogenase activity.