Dual-Energy Parathyroid 4D-CT: Improved Discrimination of Parathyroid Lesions from Thyroid Tissue Using Noncontrast 40-keV Virtual Monoenergetic Images.

BACKGROUND AND PURPOSE: In parathyroid CT, a noncontrast phase aids discrimination of parathyroid lesions (not iodine-containing) from thyroid tissue (iodine-containing). When thyroid iodine is pathologically diminished, this differentiation is difficult with standard CT. Because the attenuation of an element is maximal near its K-edge (iodine = 33.2 keV), we hypothesized that dual-energy CT 40-keV virtual monoenergetic images will accentuate thyroid iodine relative to standard images, improving the differentiation of thyroid from parathyroid lesions. Our purpose was to test this hypothesis through quantitative assessment of Hounsfield unit attenuation and contrast-to-noise on dual-energy CT standard (70-keV) and 40-keV noncontrast images. MATERIALS AND METHODS: For this retrospective study including 20 dual-energy parathyroid CTs, we used an ROI-based analysis to assess the attenuation of thyroid tissue, parathyroid lesions, and sternocleidomastoid muscle as well as corresponding contrast-to-noise on standard and 40- keV noncontrast images. Wilcoxon signed rank tests were performed to compare differences between 70 and 40 keV. RESULTS: Absolute and percentage increases in attenuation at 40 keV were significantly greater for thyroid gland than for parathyroid lesions and sternocleidomastoid muscle (P < .001 for all). Significant increases in the contrast-to-noise of thyroid relative to parathyroid lesions (median increase, 0.8; P < .001) and relative to sternocleidomastoid muscle (median increase, 1.3; P < .001) were observed at 40 keV relative to 70 keV. CONCLUSIONS: Forty-kiloelectron volt virtual monoenergetic images facilitate discrimination of parathyroid lesions from thyroid tissue by significantly increasing thyroid attenuation and associated contrast-to-noise. These findings are particularly relevant for parathyroid lesions that exhibit isoattenuation to the thyroid on parathyroid CT arterial and venous phases and could, therefore, be missed without the noncontrast phase.

Can Assessment of the Tongue on Brain MRI Aid Differentiation of Seizure from Alternative Causes of Transient Loss of Consciousness?

BACKGROUND AND PURPOSE: Transient loss of consciousness is commonly evaluated in the emergency department. Although typically caused by epileptic seizure, syncope, or psychogenic nonepileptic seizure, the underlying etiology is frequently misdiagnosed. Lateral tongue bites are reportedly a specific clinical finding of seizure. We have observed tongue signal abnormality suggesting bite injury on brain MR imaging after seizures. We hypothesized an association between tongue signal abnormality and seizure diagnosis among patients in the emergency department imaged for transient loss of consciousness. Our purposes were to determine the prevalence of tongue signal abnormality among this population and the predictive performance for seizure diagnosis. MATERIALS AND METHODS: For this retrospective study including 82 brain MR imaging examinations, 2 readers independently assessed tongue signal abnormality on T2-weighted and T2-weighted FLAIR images. Discrepancies were resolved by consensus, and interrater reliability (Cohen κ) was calculated. The final diagnosis was recorded. Proportions were compared using the Fisher exact test. RESULTS: Tongue signal abnormality was present on 19/82 (23%) MR imaging examinations. Interrater reliability was "substantial" (κ = 0.77). Seizure was diagnosed among 18/19 (95%) patients with tongue signal abnormality and 29/63 (46%) patients without it (P < .001). In our cohort, tongue signal abnormality conveyed 97% specificity, 95% positive predictive value, and 63% accuracy for seizure diagnosis. CONCLUSIONS: Tongue signal abnormality was observed in 23% of the study cohort and conveyed 97% specificity and 95% positive predictive value for seizure diagnosis. By assessing and reporting tongue signal abnormality, radiologists may facilitate a timely and accurate diagnosis of seizure among patients imaged for transient loss of consciousness.

The Pharyngolaryngeal Venous Plexus: A Potential Pitfall in Surveillance Imaging of the Neck.

BACKGROUND AND PURPOSE: Among patients undergoing serial neck CTs, we have observed variability in the appearance of the pharyngolaryngeal venous plexus, which comprises the postcricoid and posterior pharyngeal venous plexuses. We hypothesize changes in plexus appearance from therapeutic neck irradiation. The purposes of this study are to describe the CT appearance of the pharyngolaryngeal venous plexus among 2 groups undergoing serial neck CTs-patients with radiation therapy-treated laryngeal cancer and patients with medically treated lymphoma-and to assess for changes in plexus appearance attributable to radiation therapy. MATERIALS AND METHODS: For this retrospective study of 98 patients (49 in each group), 448 contrast-enhanced neck CTs (222 laryngeal cancer; 226 lymphoma) were assessed. When visible, the plexus anteroposterior diameter was measured, and morphology was categorized. RESULTS: At least 1 plexus component was identified in 36/49 patients with laryngeal cancer and 37/49 patients with lymphoma. There were no statistically significant differences in plexus visibility between the 2 groups. Median anteroposterior diameter was 2.1 mm for the postcricoid venous plexus and 1.6 mm for the posterior pharyngeal venous plexus. The most common morphology was "bilobed" for the postcricoid venous plexus and "linear" for the posterior pharyngeal venous plexus. The pharyngolaryngeal venous plexus and its components were commonly identifiable only on follow-up imaging. CONCLUSIONS: Head and neck radiologists should be familiar with the typical location and variable appearance of the pharyngolaryngeal plexus components so as not to mistake them for neoplasm. Observed variability in plexus appearance is not attributable to radiation therapy.

Internal Auditory Canal Diverticula among Pediatric Patients: Prevalence and Assessment for Hearing Loss and Anatomic Associations.

BACKGROUND AND PURPOSE: Internal auditory canal diverticula are focal lucencies along the anterior-inferior aspect of the internal auditory canal fundus. Studies in adults report conflicting data on the etiology and clinical relevance of this finding. We would expect a pediatric study to help elucidate the significance of internal auditory canal diverticula. The primary goals of this study were to determine the temporal bone CT prevalence of diverticula among pediatric patients and to assess possible hearing loss and anatomic associations. MATERIALS AND METHODS: For this retrospective study including 283 pediatric temporal bone CTs, 4 neuroradiologists independently assessed for diverticula. Discrepancies were resolved by consensus. One neuroradiologist assessed for an enlarged vestibular aqueduct, labyrinthine dysplasia, cochlear cleft, and otospongiosis. Patient demographics, audiologic data, and pertinent clinical history were recorded. One-way analysis of variance and the Fisher exact test were used to assess possible associations between diverticula and specific patient characteristics. RESULTS: Diverticula were observed in 42/283 patients (14.8%) and were more commonly bilateral. There was no significant association with age, sex, hearing loss, enlarged vestibular aqueduct, labyrinthine dysplasia, or cochlear cleft. A statistically significant association was observed with otospongiosis (P = .013), though only 1 study patient had this disease. CONCLUSIONS: Internal auditory canal diverticula are a common finding on pediatric temporal bone CT. In the absence of clinical or imaging evidence for otospongiosis, diverticula likely fall within the range of a normal anatomic variation. Familiarity with these findings may prevent neuroradiologists from recommending unnecessary additional testing in pediatric patients with isolated internal auditory canal diverticula.

Using Model-Based "Learn and Confirm" to Reveal the Pharmacokinetics-Pharmacodynamics Relationship of Pembrolizumab in the KEYNOTE-001 Trial.

Evaluation of pharmacokinetic/pharmacodynamic (PK/PD) properties played an important role in the early clinical development of pembrolizumab. Because analysis of data from a traditional 3 + 3 dose-escalation design revealed several critical uncertainties, a model-based approach was implemented to better understand these properties. Based on anticipated scenarios for potency and PK nonlinearity, a follow-up study was designed and thoroughly evaluated. Execution of 14,000 virtual trials led to the selection and implementation of a robust design that extended the low-dose range by 200-fold. Modeling of the resulting data demonstrated that pembrolizumab PKs are nonlinear at <0.3 mg/kg every 3 weeks, but linear in the clinical dose range. Saturation of ex vivo target engagement in blood began at ≥1 mg/kg every 3 weeks, and a steady-state dose of 2 mg/kg every 3 weeks was needed to reach 95% target engagement, supporting examination of 2 mg/kg every 3 weeks in ongoing trials in melanoma and other advanced cancers.

Sitagliptin 100 mg daily effect on DPP-4 inhibition and compound-specific glycemic improvement.

OBJECTIVE: In clinical trials, the degree of glucose lowering with sitagliptin has been correlated with the magnitude of dipeptidyl peptidase-4 (DPP-4) inhibition over 24 h. Previous studies evaluating sitagliptin doses ranging from 25 to 200 mg/day demonstrated that the daily dose of 100 mg provided maximal glucose-lowering efficacy for this compound in patients with type 2 diabetes. However, sitagliptin 200 mg once daily provided numerically greater percent plasma DPP-4 inhibition compared with 100 mg once daily. The purpose of this study was to evaluate whether sitagliptin 200 mg once daily provides greater improvement in glycemic efficacy as assessed by weighted mean glucose (WMG) over 24 h relative to sitagliptin 100 mg once daily and to relate the percent DPP-4 inhibition achieved with these doses to any between-treatment differences in glycemic efficacy. METHODS: In a double-blind crossover study, patients with type 2 diabetes (fasting plasma glucose [FPG] 130-250 mg/dL) were randomized to one of six treatment sequences over three treatment periods (placebo, sitagliptin 100 mg once daily, or sitagliptin 200 mg once daily). Each of the treatment periods was 7 days in duration, with 28-day washout periods between treatments. After each treatment period, patients underwent blood sampling at various time points over 24 h to determine 24-h WMG. Plasma DPP-4 activity was assessed at trough, 24 h following dosing on day 7; percent DPP-4 inhibition was corrected for sample assay dilution. RESULTS: The 103 randomized patients had a baseline mean FPG of 172 mg/dL. Following a planned interim analysis, the study was stopped because the 24-h WMG values were not different between the sitagliptin doses. Furthermore, a significant carryover effect across periods was observed for FPG; thus, efficacy results from period 1 are presented herein. The 24-h WMG values were significantly (p < 0.01) lower with sitagliptin relative to placebo, but the difference between sitagliptin doses was not significant (p = 0.365). Corrected percent plasma DPP-4 inhibition at trough was not significantly (p = 0.791) different with sitagliptin 200 mg (LS mean [95% CI] 96.9% [90.0, 100.0]) compared with sitagliptin 100 mg (95.6% [88.4, 100.0]). The early termination and the carryover effect described above are limitations to this study. CONCLUSION: Across sitagliptin doses in this study, the similarity of the 24-h WMG concentrations and the similarity of the corrected DPP-4 inhibition values support prior findings that the maximal glucose-lowering efficacy of sitagliptin is achieved with once-daily dosing of 100 mg. Clinicaltrials.gov: NCT00541229.

Structure of Na+,K+-ATPase at 11-A resolution: comparison with Ca2+-ATPase in E1 and E2 states.

Na+,K+-ATPase is a heterodimer of alpha and beta subunits and a member of the P-type ATPase family of ion pumps. Here we present an 11-A structure of the heterodimer determined from electron micrographs of unstained frozen-hydrated tubular crystals. For this reconstruction, the enzyme was isolated from supraorbital glands of salt-adapted ducks and was crystallized within the native membranes. Crystallization conditions fixed Na+,K+-ATPase in the vanadate-inhibited E2 conformation, and the crystals had p1 symmetry. A large number of helical symmetries were observed, so a three-dimensional structure was calculated by averaging both Fourier-Bessel coefficients and real-space structures of data from the different symmetries. The resulting structure clearly reveals cytoplasmic, transmembrane, and extracellular regions of the molecule with densities separately attributable to alpha and beta subunits. The overall shape bears a remarkable resemblance to the E2 structure of rabbit sarcoplasmic reticulum Ca2+-ATPase. After aligning these two structures, atomic coordinates for Ca2+-ATPase were fit to Na+,K+-ATPase, and several flexible surface loops, which fit the map poorly, were associated with sequences that differ in the two pumps. Nevertheless, cytoplasmic domains were very similarly arranged, suggesting that the E2-to-E1 conformational change postulated for Ca2+-ATPase probably applies to Na+,K+-ATPase as well as other P-type ATPases.

Ligands presumed to label high affinity and low affinity ATP binding sites do not interact in an (alpha beta)2 diprotomer in duck nasal gland Na+,K+-ATPase, nor Do the sites coexist in native enzyme.

The interaction of ligands deemed to be ATP analogues with renal Na(+),K(+)-ATPase suggests that two ATP binding sites coexist on each functional unit. Previous studies in which fluorescein 5-isothiocyanate (FITC) was used to label the high affinity ATP site and 2'(3')-O-(2,4,6-trinitrophenyl)adenosine 5'-diphosphate (TNP-ADP) was used to probe the low affinity site suggested that the two sites coexist on the same alphabeta protomer. Other studies in which FITC labeled the high affinity site and erythrosin-5-isothiocyanate (ErITC) labeled the low affinity site led to the conclusion that the high and low affinity sites exist on separate interacting protomers in a functional diprotomer. We report here that at 100% inhibition of ATPase activity by FITC, each alphabeta protomer of duck nasal gland enzyme has a single bound FITC. Both TNP-ADP and ErITC interact with FITC-bound protomers, which unambiguously demonstrates that putative high and low affinity ATP sites coexist on the same protomer. In unlabeled nasal gland enzyme, TNP-ADP and ErITC inhibit both ATPase activity and p-nitrophenyl phosphatase activity, functions attributed to the putative high and low affinity ATP site, respectively, by interacting with a single site with characteristics of the high affinity ATP binding site. In FITC-labeled enzyme, TNP-ADP and ErITC inhibit p- nitrophenyl phosphatase activity but at much higher concentrations than with the unmodified enzyme. Low affinity sites do not exist on the unmodified enzyme but can be detected only after the high affinity site is modified by FITC.

Alphabeta protomers of Na+,K+-ATPase from microsomes of duck salt gland are mostly monomeric: formation of higher oligomers does not modify molecular activity.

The distance that separates alphabeta protomers of the Na(+), K(+)-ATPase in microsomes and in purified membranes prepared from duck nasal salt glands was estimated by measuring fluorescence resonance energy transfer between anthroylouabain bound to a population of alphabeta protomers and either N-[7-nitrobenz-2-oxa-1, 3-diazol-4-yl]-6-aminohexyl ouabain or 5-(and-6)-carboxyfluorescein-6-aminohexyl ouabain bound to the rest. Energy transfer between probes bound in the microsomal preparation was less than in the purified membranes. The efficiency of energy transfer between anthroylouabain and N-[7-nitrobenz-2-oxa-1, 3-diazol-4-yl]-6-aminohexyl ouabain was 29.2% in the microsomes compared with 62.6% in the purified preparation. Similar results were obtained with 5-(and-6)-carboxyfluorescein-6-aminohexyl ouabain as acceptor. We calculate that either the protomer bound probes were on the average 13 A farther apart in the microsomes than in the purified membranes, or that 53% of the protomers are monomeric in the microsome preparation. Microsomes prepared in the presence of phalloidin (a toxin that binds to F actin and stabilizes the actin-based cytoskeleton) showed less quench than those prepared in its absence. The data support the hypothesis that protomers are kept apart by their association with the cytoskeleton. The turnover rate while hydrolyzing ATP is the same in the microsomal and purified preparations; higher oligomer formation has no significant effect on the enzyme reaction mechanism.

Role of polyamine structure in inhibition of K+-Cl- cotransport in human red cell ghosts.

1. K+-Cl- cotransport in human red cell ghosts is inhibited by divalent inorganic cations, soluble polycations and amphipathic organic cations. These findings suggest a common mechanism of inhibition, namely, binding of the cations to negative charges at the surface of a hydrophobic structure. 2. We have characterized the inhibitory capacity of a number of polyamines in order to obtain information about the nature of the charges with which they interact. Neomycin inhibited swelling-stimulated cotransport. The diquaternary amines dimethonium and decamethonium were relatively ineffective inhibitors. These compounds are thought to shield negative charges, but not bind to them. 3. Comparison of a homologous series of polyamines indicated that primary amines were better inhibitors than secondary amines, that inhibition increased with the charge of the polyamine, and that inhibition increased as the distance separating the amines increased. 4. The results indicate that the negative charges to which polycations bind are multiple and mobile. Since they must be associated with a hydrophobic environment, it is likely that they are negatively charged phospholipids located in the inner leaflet of the bilayer membrane. 5. Heating red cells or ghosts to 49 C denatures spectrin. Heating markedly increased K+ uptake in swollen ghosts but not in shrunken ghosts. The increase in uptake was reversed when swollen ghosts were shrunk even though denaturation of spectrin was not reversed. Polyamines, which inhibited swelling-activated K+ uptake in control ghosts, similarly inhibited the increased uptake in heated ghosts. 6. We speculate that spectrin, which is closely associated with the inner bilayer leaflet, shields negative charges in a volume-dependent manner and so regulates volume-sensitive K+ transport.

Preparation of Na+,K+-ATPase with near maximal specific activity and phosphorylation capacity: evidence that the reaction mechanism involves all of the sites.

The phosphorylation capacity of Na+,K+-ATPase preparations in common use is much less than expected on the basis of the molecular weight of the enzyme deduced from cDNA sequences. This has led to the popularity of half-of-the-sites or flip-flop models for the enzyme reaction mechanism. We have prepared Na+,K+-ATPase from nasal salt glands of salt-adapted ducks which has a phosphorylation capacity and specific activity near the theoretical maxima. Preparations with specific activities of >60 micromol (mg of protein)-1 min-1 at 37 degrees C had phosphorylation capacities of >60 nmol/mg of protein, and the rate of turnover of the enzyme was 9690 min-1, within the range reported for the enzyme from other sources. The fraction of the maximal specific activity of the enzyme compared well with the fraction of the protein on SDS-PAGE which was alpha and beta chains, especially at the highest specific activity which indicates that all of the alphabeta protomers are active. The gels of the most reactive preparations contained only alpha and beta chains, but less active preparations contained a number of extraneous proteins. The major contaminant was actin. The preparation did not contain any protein which migrated in the molecular weight range of the gamma subunit. The subunit composition of the enzyme was alpha1 and beta1 only. This is the first report of a pure, homogeneous, fully active preparation of the protein. Reaction models which incorporate a half-of-the-sites or flip-flop mechanism do not apply to this enzyme.

Electromechanical spectroscopy of cartilage using a surface probe with applied mechanical displacement.

This study focuses on an approach for the nondestructive assessment of cartilage degeneration in vivo by quantitation of bulk material properties based on measurements made at the top surface of the tissue. A model of an electromechanical coupled poroelastic medium [Sachs and Grodzinsky (1989) Physicochem. Hydrodyn. 11, 585-614] is used to interpret the behavior of a diagnostic probe configuration suitable for such surface measurements of cartilage electromechanical and poroelastic properties via arthroscopy. The response of a planar layer of tissue to a periodic displacement imposed at the articular surface is described. This displacement produces a periodic electric streaming potential and mechanical stress in the bulk and at the surface of the tissue layer with the same frequency and wavelength as the imposed displacement. Using boundary conditions and parameter values relevant to cartilage, the results show that surface measurements of the stress and potential can be used to determine bulk material properties including tissue thickness, moduli, hydraulic permeability, and electrokinetic coupling coefficients. The relation between the temporal frequency and spatial wavelength of the surface excitation and the amplitude, phase, and penetration depth of the stress and potential is investigated numerically and asymptotically. Good agreement has been found between the long wave limit (with parameter values taken from the literature) and compression-induced streaming potential data from previous experiments in uniaxial confined compression. The results show that use of independently imposed temporal frequency and spatial wavelength may enable detection and imaging of focal regions of cartilage degeneration via nondestructive surface spectroscopy.

The role of (alpha beta) protomer interaction in determining functional characteristics of red cell Na,K-ATPase.

We have examined the possibility that interaction of (alpha beta) protomers within a diprotomer is responsible for some anomalous characteristics of red cell Na,K-ATPase by examining their response to two inhibitors, FITC and H2DIDS, which bind covalently, and to ouabain, which debinds slowly from red cell pumps. The phenomena we examined were: (1) the biphasic curve relating Na,K-ATPase activity to ATP concentration, and (2) protection of Na pumps against vanadate inhibition by external Na. If interaction of (alpha beta) protomers within a diprotomer were responsible for these phenomena, random inactivation of (alpha beta) protomers should have resulted in a high proportion of (alpha beta) promtomers with an inhibited protomer as a partner, and therefore should have significantly altered the consequences of subunit interaction. With each inhibitor, 60-70% inhibition of ATPase activity did not alter the functional characteristics of the residual activity. We conclude that interaction of functional (alpha beta) protomers does not explain the phenomena which we investigated. This is consistent with our previous observation that Na,K pumps of red cell membranes exist as monomeric (alpha beta) protomers (Martin, D.W. and Sachs, V.R. (1992) J. Biol. Chem. 267, 23922-23929).

Soluble polycations and cationic amphiphiles inhibit volume-sensitive K-Cl cotransport in human red cell ghosts.

We have measured the effect of soluble polycations (spermine and methylglyoxal) and cationic amphiphiles (sphingosine and tetracaine) on K-Cl cotransport in shrunken and swollen red cell ghosts. All substances inhibited cotransport, and for each agent, the concentration at which inhibition was half-maximal was about the same for swollen and shrunken ghosts. Acetylspermine was a much less effective inhibitor than spermine, which demonstrates that inhibition depends on the cationic groups of spermine. Spermine was a more effective inhibitor in ATP-free ghosts than in ghosts containing ATP, which eliminates the possibility that inhibition of cotransport activity results from inhibition of protein kinase activity. Inhibition by spermine is as effective in K-free ghosts as in high-K ghosts; spermine does not inhibit cotransport by reducing the effective K concentration at the inner membrane surface. We conclude that regulation of K-Cl cotransport involves negative charges (phosphatidylserine or phosphatidylinositides) at the inner membrane surface and suggest a model that accounts for our findings.

The role of ATP in swelling-stimulated K-Cl cotransport in human red cell ghosts. Phosphorylation-dephosphorylation events are not in the signal transduction pathway.

Volume-sensitive K-Cl cotransport occurs in red blood cells of many species. In intact cells, activation of K-Cl cotransport by swelling requires dephosphorylation of some cell protein, but maximal activity requires the presence of intracellular ATP. We have examined the relation between K-Cl cotransport activity and ATP in ghosts prepared from human red blood cells. K-Cl cotransport activity in swollen ghosts increased by ATP, and the increase requires Mg so that it almost certainly results from the phosphorylation of some membrane component. However, even in ATP-free ghosts residual volume-sensitive K-Cl cotransport can be demonstrated. This residual cotransport in ATP-free ghosts is greater in the presence of vanadate, a tyrosyl phosphatase inhibitor, and in ghosts that contain ATP cotransport is reduced by genistein, a tyrosyl kinase inhibitor. Okadaic acid, an inhibitor of serine and threonine phosphatases, inhibits K-Cl cotransport in ghosts as it does in intact cells. Experiments in which ghosts were preexposed to okadaic acid showed that the protein dephosphorylation that permits K-Cl cotransport can proceed to completion before the ghosts are swollen and K transport measured and therefore dephosphorylation is not a response to ghost swelling. In experiments with ATP-free ghosts we found that phosphorylation is not necessary to increase the cotransport rate when shrunken ghosts are swollen, nor is rephosphorylation necessary to decrease the cotransport rate when swollen ghosts are shrunken. Cotransport is greater in swollen than in shrunken ghosts even when the swollen and shrunken ghosts have the same concentration of cytoplasmic solutes. We conclude that, although phosphorylation and dephosphorylation modify the activity of the cotransporter in swollen and in shrunken ghosts, neither of these processes nor any other known messenger is involved in signal transduction between the cell volume sensor and the cotransporter as originally proposed by Jennings and Al-Rohil (Jennings, M. L., and N. Al-Rohil. 1990. Journal of General Physiology. 95: 1021-1040).

Cross-linking of the erythrocyte (Na+,K+)-ATPase. Chemical cross-linkers induce alpha-subunit-band 3 heterodimers and do not induce alpha-subunit homodimers.

Earlier studies (Periyasamy, S. M., Huang, W.-H., and Askari, A. (1983) J. Biol. Chem. 258, 9878-9885) suggested that Cu2+ and o-phenanthroline induced the formation of cross-linked homodimers between alpha-subunits of the erythrocyte (Na+,K+)-ATPase. This was interpreted as indicating that alpha-subunits existed in close proximity in native erythrocyte membranes. The alpha-subunit and band 3 monomers have similar molecular weights (M(r) approximately 100,000) and exist in the membrane in molar ratios of approximately 1:3000 alpha-subunit:band 3. We explored the possibility that alpha-subunit and band 3 could be induced to form heterodimeric structures in the presence of cross-linking reagents. Using methods similar to those employed in the above-cited reference we demonstrated that cross-linked dimers containing phosphorylated alpha-subunits had proteolytic sensitivity that was inconsistent with the formation of alpha-subunit homodimers and fully consistent with heterodimer formation between alpha-subunit and band 3. The data also indicated that alpha-subunit-band 3 heterodimer formation is dependent on the conformational state of the (Na+,K+)-ATPase. Using the appropriate reagents we obtained cross-linked products which were consistent with heterodimer formation between alpha- and beta-subunits of the (Na+,K+)-ATPase. Our data argue against a close association between pairs of (Na+,K+)-ATPase alpha-subunits in the human red cell membrane.

Perialveolar interstitial resistance and compliance in isolated rat lung.

We have developed a method to characterize fluid transport through the perialveolar interstitium using micropuncture techniques. In 10 experiments we established isolated perfused rat lung preparations. The lungs were initially isogravimetric at 10 cmH2O arterial pressure, 2 cmH2O venous pressure, and 5 cmH2O alveolar pressure. Perialveolar interstitial pressure was determined by micropuncture at alveolar junctions by use of the servo-null technique. Simultaneously a second micropipette was placed in an alveolar junction 20-40 microns away, and a bolus of albumin solution (3.5 g/100 ml) was injected. The resulting pressure transient was recorded for injection durations of 1 and 4 s in nonedematous lungs. The measurements were repeated after gross edema formation induced by elevated perfusion pressure. We model the interstitium as a homogeneous linearly poroelastic material and assume the initial pressure distribution due to the injection to be Gaussian. The pressure decay is inversely proportional to time, with time constant T, where T is a measure of the ratio of interstitial tissue stiffness to interstitial resistance to fluid flow. A linear regression was performed on the reciprocal of the pressure for the decaying portion of the transients to determine T. Comparing pressure transients in nonedematous and edematous lungs, we found that T was 4.0 +/- 1.4 and 1.4 +/- 0.6 s, respectively. We have shown that fluid transport through the pulmonary interstitium on a local level is sensitive to changes in interstitial stiffness and resistance. These results are consistent with the decreased stiffness and resistance in the perialveolar interstitium that accompany increased hydration.

Disseminated Pneumocystis carinii infection with hepatic involvement in a patient with the acquired immune deficiency syndrome.

Extrapulmonary infection with Pneumocystis carinii (P. carinii) in AIDS patients is uncommon, and is often described only at postmortem examination. Although most antemortem cases involve spread to the bone marrow or spleen, P. carinii involvement of other organs has only recently been described. Despite the frequency of liver enzyme abnormalities in AIDS patients with a history of P. carinii pneumonia, P. carinii has been observed only rarely in the liver. We present a well-documented case of P. carinii involving the liver in an AIDS patient with P. carinii pneumonia and progressive liver enzyme abnormalities. We suggest that P. carinii infection should be considered in the differential diagnosis of AIDS-related liver disease.