Screening head ultrasound to detect intraventricular hemorrhage in premature infants.

OBJECTIVE: The objective of this case-control study was to develop a screening protocol using head ultrasound (HUS) to detect high-grade intraventricular hemorrhage (IVH) in very-low-birthweight infants with greater specificity than current practice, while maintaining a high degree of sensitivity. MATERIALS AND METHODS: All infants /= 10, or cardiopulmonary resuscitation in the neonatal intensive care unit. CONCLUSION: Infants born at 28-32 weeks with a high-grade IVH can be identified with a high degree of sensitivity using refined screening criteria, eliminating 50 % of the HUS scans currently obtained for IVH screening.

Rectal methohexital for sedation of children during imaging procedures.

OBJECTIVE: Brevital (methohexital), administered as a solution per rectum, provides safe, rapid, and effective sedation for induction of surgical anesthesia. This study was undertaken to evaluate rectal Brevital as a sedative for children undergoing imaging procedures. MATERIALS AND METHODS: In a review of patients' records, we found that, during a 1-year period, 190 children from 1 month to 14 years old were sedated for CT or MR imaging. Sedation was required for 94 CT and 96 MR imaging studies. Rectal Brevital was used in 102 patients (mean age, 25 +/- 2 months), and oral chloral hydrate was used in 88 (mean age, 28 +/- 3 months). RESULTS: Sleep was achieved in 81% of patients who received Brevital and in 80% of those who received chloral hydrate. Induction time was shorter (p = .0001) with Brevital (9 +/- 1 min) than with chloral hydrate (28 +/- 2 min). The mean duration of sleep was 46 min with Brevital and 66 min with chloral hydrate (p = .0001). Brevital provided adequate sedation in 89 (87%) of 102 imaging studies, and chloral hydrate did so in 73 (83%) of 88 studies. No cardiorespiratory complications or allergic reactions occurred with either drug. Significantly fewer patients were discharged fully awake and alert after sedation with chloral hydrate than with Brevital (p < .002). Children sedated with chloral hydrate also required a longer period of observation in the radiology department (p < .04). CONCLUSION: We conclude that rectal Brevital produces sedation of adequate duration for most imaging procedures in children. Ease of administration, wide margin of safety, rapid and pleasant induction, and short recovery time make this drug a favorable alternative to other commonly used sedatives.

Extracellular pH modifies adaptive response to high K+ in cultured canine kidney cells.

The chronic interactive and independent effects of extracellular pH and K+ on renal Na(+)-K(+)-adenosinetriphosphatase (ATPase) activity and active K+ transport were studied in the Madin-Darby canine kidney (MDCK) cell line. Confluent cell monolayers were incubated for 24 h in control (4 mM) or high (7.5 mM) K+ medium at acid (6.8) or neutral (7.4) pH. Under acid pH conditions, exposure to high K+ elicited a rise of 133% in maximum Na(+)-K(+)-ATPase activity and 66% in active K+ uptake. In contrast, high K+ had no effect on enzyme activity or K+ uptake at neutral pH. Detergent-activated Na(+)-K(+)-ATPase assay demonstrated a latent pool of enzyme at acid pH-control K+, which seemed to account entirely for the increase in Na(+)-K(+)-ATPase activity after exposure to high K+. The effects of pH appeared unrelated to HCO3- and Cl- concentration in the extracellular environment. We conclude that the upregulatory effect of high K+ on renal Na(+)-K(+)-ATPase is pH dependent. The data suggest that a pool of catalytically inactive enzyme exists only at acid extracellular pH at K+ concentrations in the normal physiological range and that K+ adaptation, at least initially, is the result of recruitment of this latent intracellular pool. In the intact cell extracellular K+ and luminal pH may interact to modify catalytic turnover rate as well as bioavailability of Na(+)-K(+)-ATPase.

The role of cortical Na,K-ATPase in distal nephron potassium secretion by the immature canine kidney.

Amiloride-sensitive potassium secretion in response to acute potassium loading is lower in the newborn than in the adult. Potassium secretion is a function of late distal tubule and cortical collecting tubule Na,K-ATPase activity. Na,K-ATPase activity in vivo is determined by enzyme abundance and catalytic turnover. Chronic potassium supplementation increases potassium secretory capacity in the adult by increasing enzyme abundance in the late distal and cortical collecting tubules. We hypothesized that the lower potassium secretory capacity of the newborn was the result of lower late distal and cortical collecting tubule Na,K-ATPase activity and could be similarly enhanced. To test this hypothesis, newborn dogs were supplemented with 6 mmol KCl.d-1.kg-1 for 1 wk; age-matched litter mate controls were not (n = 8 pairs). Potassium supplementation resulted in a mean increase in Vmax Na,K-ATPase activity in vitro (proportional to pump abundance) of 70 +/- 42%. Mean Na,K-ATPase activities +/- SEM were 279 +/- 58 versus 198 +/- 44 nmol inorganic P. h-1.microgram DNA-1, p = 0.05. However, amiloride-sensitive potassium secretion after an acute potassium load of 20 mumol.min-1.kg-1 over 150 min was not enhanced (9.6 +/- 1.8 versus 8.9 +/- 0.8 mumol.min-1.kg-1, potassium-supplemented versus control animals). We conclude that lower enzyme abundance is not primarily responsible for the newborn's lower potassium secretory capacity. We speculate that the factor that limits secretion in the newborn during acute potassium loading does so by restricting catalytic turnover of the enzyme in vivo.

Na(+)-K(+)-ATPase in adipocyte differentiation in culture.

Differentiation of 3T3-L1 cells from a fibroblast to an adipocyte phenotype results in an approximately 50% decline in Na(+)-K(+)-ATPase activity and ouabain-sensitive 86Rb uptake. Kinetic analysis revealed a K 1/2 for Na+ of approximately 14 mM, a Km for ATP of approximately 0.4 mM, and maximal activation by sodium dodecyl sulfate at a 0.05 (wt/wt) detergent/protein ratio in both mature fibroblasts and adipocytes. Both fibroblasts and adipocytes exhibited Na(+)-K(+)-ATPase activity with an inhibition constant (Ki) for ouabain of approximately 10(-4) M. In addition, adipocytes exhibited a second component representing 30% of total activity with a Ki of approximately 5 x 10(-7) M. The emergence of biphasic ouabain inhibition kinetics in adipocytes raised the possibility of a change in alpha-subunit isoform composition with cytodifferentiation. This inference was evaluated by isoform-specific mRNA analysis (Northern blots) and by alpha-isoform-specific immunoassays (Western blots). Northern blots revealed a modest decrease in mRNA alpha 1, a striking increase in mRNA alpha 2, and a significant loss of mRNA beta content with differentiation of fibroblasts to adipocytes. By immunoassay, fibroblasts exhibited the alpha 1-isoform. Adipocytes exhibited an admixture of alpha 1- and alpha 2-isoforms, with alpha 2 being the more abundant isoform. There was no one-to-one correspondence either between the mRNA isoform and alpha-subunit abundances or between alpha-subunit abundances and enzymatic activity, suggesting that regulation occurs at multiple levels in this system. Findings indicate, however, that a shift in alpha-isoform composition accompanied by a change in ouabain inhibition kinetics occurs with cytodifferentiation.

Effect of high extracellular K+ on Na-K-ATPase in cultured canine kidney cells.

The Madin-Darby canine kidney (MDCK) cell line was used to evaluate the influence of high extracellular K+, independent of hormonal effects, on renal Na-K-adenosinetriphosphatase (ATPase) activity and abundance. Confluent cell monolayers were incubated in control (5 mM) or high K+ (7.5 mM) medium for 24 h. Exposure to high K+ elicited a 46% rise in Na-K-ATPase activity and a 55% increase in ouabain-sensitive 86Rb uptake. Na-K-ATPase abundance, estimated from the number of ouabain-binding sites, also increased 63% over control in cells exposed to 7.5 mM K+, and as a consequence there was no statistically significant change in the catalytic turnover number. Northern blot analysis using rat cDNA probes for the alpha 1- and beta-subunits showed no corresponding changes in subunit-specific mRNA abundances at 24 h. We conclude that chronic exposure to high extracellular K+ produces a rise in renal epithelial Na-K-ATPase activity and active K+ transport, independent of changes in aldosterone, renal blood flow, or extracellular Na+ concentration. This effect is due to an increase in enzyme abundance rather than a change in catalytic turnover rate. The results of Northern analysis suggest that regulation of Na-K-ATPase activity and abundance by high K+ may involve translational or posttranslational mechanisms, but further study with cDNA probes of canine origin is needed to resolve this issue.

Chronic potassium supplementation of newborn dogs increases cortical Na,K-ATPase but not urinary potassium excretion.

The distal nephron of the newborn dog cannot secrete an acute potassium load as efficiently as can that of the adult dog. Distal nephron potassium secretion is dependent upon basolateral Na,K-ATPase activity. Because Na,K-ATPase activity is lower in the immature than the mature distal nephron, it was hypothesized that lower Na,K-ATPase activity may be responsible for the lower potassium secretory capacity of the immature nephron. In the adult, chronic high dietary potassium intake increases renal tubular potassium secretory capacity by increasing Na/K pump abundance in distal nephron segments responsible for potassium secretion. Therefore, in order to test the above hypothesis, renal cortical and outer medullary Na,K-ATPase activity under Vmax conditions (a measure of pump abundance) and urinary potassium excretion during acute potassium loading were determined in 7 age-matched, litter mate pairs (chronically potassium supplemented versus control) newborn dogs. The potassium supplemented member of each pair received 6 mmol.day-1.kg-1 of KCl as a 150 mM solution for 7-21 days after birth and the control member received an equal volume of water for the same period of time. This protocol resulted in a doubling of renal cortical Vmax Na,K-ATPase activity in the potassium supplemented animals (from 369 +/- 186 to 718 +/- 286 nmol Pi liberated.h-1.micrograms DNA-1, P = 0.025). There was no significant change in outer medullary enzyme activity. Contrary to the above hypothesis, this increase in cortical enzyme activity was not associated with increased potassium excretion at baseline or during acute potassium loading.(ABSTRACT TRUNCATED AT 250 WORDS)