What questions do patients with curable prostate cancer want answered?

PURPOSE: To determine the questions that recently diagnosed early-stage prostate cancer patients think should be addressed with patients like themselves. STUDY POPULATION: 56 patients diagnosed as having early-stage prostate cancer within the previous year. METHODS: Surveys distributed to the patients included 93 questions that might be considered important. Respondents judged the importance (essential/desired/no opinion/avoid) of addressing each question, and indicated why those "essential" or "desired" were important. RESULTS: 38 patients (68%) responded. Agreement on question importance, overall, was rather poor (mean 41.6%, kappa 0.17). There were, however, 20 questions that at least 67% of the respondents agreed were essential to address and 12 that they agreed were not essential. No question was relevant to the treatment decisions of more than 50% of respondents, but 91 questions were relevant to at least one respondent's decision. CONCLUSIONS: Although there was enough agreement to define a core set of questions that should be addressed with most patients, each of the remaining questions was also considered essential to some people. The core set, therefore, would not be adequate to satisfy any one patient's essential information needs. Whereas most questions would be needed to cover all patients' decision needs, only some are needed for any given patient. Such variability in information needs means that the subjective standard is the only viable legal standard for judging the adequacy of provision of information for the treatment decision.

Retention of nociceptor responses during deep barbiturate anesthesia in frogs.

Bullfrogs (Rana catesbeiana) anesthetized with a large dose of thiopental (42.8 mg/kg) retained movement responses to nociceptor stimuli despite an average plasma drug level of 51 mg/l, of which 63% was bound to plasma proteins. This concentration, when corrected to include only unbound and uncharged drug, was 2-fold greater than those reported to abolish nociceptor response (NR) during surgical anesthesia in man. The median anesthetic dose (AD50) for loss of the righting reflex was 11.2 mg/kg by s.c. injection into the abdominal lymph sac; however, at 54.0 mg/kg, all frogs retained NRs, although otherwise deeply anesthetized. The ratio of NR-blocking dose to light AD was thus > 4.8, as compared to < 2 in mammalian studies. Whole body levels of thiopental determined at 3 h after intralymphatic injection showed that about half the injected drug had been eliminated by this time and that termination of anesthesia was chiefly due to drug elimination. Even though the pharmacokinetics of thiopental appears to differ markedly in frogs and men, the poor analgesia seen in the present study frequently has been reported during clinical barbiturate anesthesia. Since this deficiency is much more pronounced in the bullfrog than in man, its neurophysiological basis might profitably be studied using the bullfrog as a model; however, the high mortality associated with deep thiopental anesthesia in the frog should preclude its use as a practical anesthetic in amphibia.

What prostate cancer patients should know: variation in professionals' opinions.

PURPOSE: This study was designed to determine what questions health-care professionals think should be addressed with curable prostate cancer patients before treatment decisions are made. METHOD: A survey was distributed to radiation oncologists, urologists, medical oncologists, nurses and radiation therapy technologists (RTTs) involved in treating prostate cancer patients. Participants were asked to judge the importance of addressing each of 78 questions (essential/important/no opinion/avoid) with a described hypothetical patient prior to the treatment decision. Eighty participants were later selected at random for a retest. RESULTS: The overall response rate was 55% (284/518) on the initial survey and 56% (45/80) on the retest. The relative importance of the various questions was similar across groups (r(76) ranged from 0.75 to 0.91, all P<0.001). Despite the between-group similarity, opinions within each group varied widely. For example, among oncologists, the number of questions deemed essential by individual respondents ranged from five to 69, with >90% respondent agreement on only 15 of the 78 questions. The extent of agreement was similar in the other groups. The retest showed that essential and important responses were reasonably stable, i.e. 92% of questions judged essential at one time were judged either essential (58%) or important (34%) at the other time. CONCLUSIONS: Although the relative importance of addressing the various questions appears similar across the professional groups involved in the care of prostate patients, within each profession there seems to be little agreement. The lack of agreement includes both how many questions are essential to address and whether or not most individual questions are essential.

Contrasting effects of anesthetics in tadpole bioassays.

The median anesthetic concentrations (AC 50 values) in the bath solution for loss of righting reflex (RR) or nociceptor response (NR) were determined in larval Rana catesbeiana. The AC50 (NR)/AC50(RR) ratios were between 1.6 and 1.9 for ethanol, ether and diazepam. Comparable ratios for hexobarbital and thiopental were 9.8 and 6.2, which indicated a marked lack of "analgesic" effect. The tadpole AC50 values were consistently higher than AC50 values reported for nonionized and unbound drug in mammalian blood water. The tadpole/mammal ratio for loss of NR during thiopental anesthesia was approximate 8, and analysis of ventricular blood showed that the ultrahigh bath concentrations of thiopental were in equilibrium with those in tadpole serum. The tadpole/mammal ratio for diazepam was approximate 110, and its effect in the tadpole was not reversed by flumazenil. Tadpole/mammal ratios for ethanol and ether were approximate 2. Ethanol and ether showed additive effects when tested in combination, and ethanol-tolerant tadpoles showed a similar tolerance to ether. In contrast, hexobarbital and ethanol showed supra-additive effects, and ethanol-tolerant tadpoles showed a greater tolerance to hexobarbital than to ethanol or ether. These studies show marked differences in effect between barbiturates and the ethanol-ether group, high-concentration benzodiazepine effects presumably independent of a specific receptor, and marked differences in drug potency between amphibia and mammals.

CHEB, a convulsant barbiturate, evokes calcium-dependent spontaneous glutamate release from rat cerebrocortical synaptosomes.

CHEB [5-(2-cyclohexylidene-ethyl)-5-ethyl barbituric acid] is a potent convulsant barbiturate that causes direct neuronal excitation by an unknown mechanism. We have analyzed the effects of CHEB on the release of endogenous glutamate from rat cerebrocortical synaptosomes using an on-line enzyme-coupled fluorimetric assay. CHEB evoked spontaneous Ca(2+)-dependent glutamate release with an EC50 = 14.2 microM and an Emax = 3.2 mumol/min/mg. The non-convulsant barbiturates pentobarbital and phenobarbital evoked significantly less glutamate release at high concentrations. CHEB (30 microM) increased intrasynaptosomal [Ca2+] by 58 +/- 4 nM (p < 0.01; n = 4) above baseline compared to an increase of 5 +/- 4 nM (NS; n = 4) produced by pentobarbital (30 microM). CHEB-evoked glutamate release was inhibited by pentobarbital, phenobarbital, EGTA, CoCl2/CdCl2 and flunarizine, but not by local anesthetics, tetrodotoxin, nitrendipine or omega-conotoxin GVIA. These results demonstrate that CHEB acts as a potent and effective secretogogue for glutamate by a pre-synaptic mechanism that does not require activation of Na+ channels or of L-type or N-type Ca2+ channels. Stimulation of spontaneous glutamate release may contribute to the convulsant properties of CHEB.

Forebrain ischemia increases GLUT1 protein in brain microvessels and parenchyma.

Glucose transport into nonneuronal brain cells uses differently glycosylated forms of the glucose transport protein, GLUT1. Microvascular GLUT1 is readily seen on immunocytochemistry, although its parenchymal localization has been difficult. Following ischemia, GLUT1 mRNA increases, but whether GLUT1 protein also changes is uncertain. Therefore, we examined the immunocytochemical distribution of GLUT1 in normal rat brain and after transient global forebrain ischemia. A novel immunocytochemical finding was peptide-inhibitable GLUT1 immunoreactive staining in parenchyma as well as in cerebral microvessels. In nonischemic rats, parenchymal GLUT1 staining co-localizes with glial fibrillary acidic protein (GFAP) in perivascular foot processes of astrocytes. By 24 h after ischemia, both microvascular and nonmicrovascular GLUT1 immunoreactivity increased widely, persisting at 4 days postischemia. Vascularity within sections of brain similarly increased after ischemia. Increased parenchymal GLUT1 expression was paralleled by staining for GFAP, suggesting that nonvascular GLUT1 overexpression may occur in reactive astrocytes. A final observation was a rapid expression of inducible heat shock protein (HSP)70 in hippocampus and cortex by 24 h after ischemia. We conclude that GLUT1 is normally immunocytochemically detectable in cerebral microvessels and parenchyma and that parenchymal expression occurs in some astroglia. After global cerebral ischemia, GLUT1 overexpression occurs rapidly and widely in microvessels and parenchyma; its overexpression may be related to an immediate early-gene form of response to cellular stress.

Muscle weakness during tricaine anesthesia.

An isolated preparation of tadpole tail muscle was used to assess the peripheral effects of tricaine (3-aminobenzoic acid ethyl ester) at anesthetic concentrations and under physiological conditions. The drug effect on the electrically-evoked twitch was tested using short-pulse durations that elicited synaptically mediated effects or longer-duration pulses that stimulated the muscle directly. Tricaine reduced both types of response anesthetic and even subanesthetic concentrations. At steady state concentrations that produced surgical anesthesia in vivo, tricaine reduced the directly evoked response by about half. It is concluded that tricaine anesthesia has a pronounced peripheral effect on neuromuscular function and that direct effect(s) on muscle are a major component.

Progressive hippocampal loss of immunoreactive GLUT3, the neuron-specific glucose transporter, after global forebrain ischemia in the rat.

Brain damage after global forebrain ischemia is worsened by prior hyperglycemia and ameliorated by antecedent hypoglycemia. To assess whether GLUT3, the neuron specific glucose transporter and its mRNA, are affected by cerebral ischemia, we investigated the hippocampal pattern of GLUT3 immunoreactivity and GLUT3 gene expression 1, 4 and 7 days after global forebrain ischemia in a rat 2-vessel occlusion model. We used a newly generated, specific, C-terminally directed polyclonal antiserum against GLUT3 to stain coronal frozen sections. Thionin staining and the microglial marker, OX42, indicated the extent of ischemic damage in hippocampus and correlated with GLUT3 loss. One day after ischemia, no significant change in hippocampal GLUT3 immunoreactivity was observed; by 4 days however, there was consistent and pronounced loss; and at 7 days the loss of GLUT3 staining was maximal. The greatest loss of GLUT3 staining was in the CA1 region, especially the strata oriens and radiatum of Ammon's horn. By contrast, GLUT3 staining was undiminished in the stratum lacunosum moleculare, in the mossy fibers of the lateral aspect of CA3 and in all but the inner-most portion of the molecular layer of the dentate gyrus, immediately adjacent to the granule cells. GLUT3 mRNA levels were not significantly altered at 24 hours and significantly declined at 4 and 7 days after ischemia in the CA1 pyramidal layer. These data are consistent with the pattern of neuronal loss and microglial activation in hippocampus. Loss of GLUT3 may affect the availability of glucose, and possibly the viability of ischemically damaged neurons.

Binding of thiopental in neonatal serum.

Protein binding of thiopental was studied in 21 samples of neonatal serum (from placental blood) and compared with protein binding in ten healthy volunteers. These infants ranged between 32 and 43 weeks of gestational age (mean, 37.7 weeks) and the adult age range was from 27 to 54 years (mean, 35.4 years). Because the unbound fraction of the drug is responsible for its pharmacologic effect, a marked difference in the protein binding between neonates and adults may be relevant to the clinician. Blood obtained from freshly delivered placentas or from adult volunteers was allowed to clot and the serum separated from the sample. A portion of the serum was sent for protein and bilirubin analysis and the remainder retained for study. This latter serum was combined with four concentrations of thiopental. These specimens were then ultrafiltered and the amount of thiopental in the ultrafiltrate (unbound) compared with the prefiltered amount (total), as measured by reverse-phase high-performance liquid chromatography. The binding studies were repeated at pH 7.2, 7.4, and 7.6 in both the adult and neonatal serum. Total protein and albumin are significantly less in neonatal serum, whereas bilirubin (total and direct) is significantly higher in neonatal serum than in adult serum (P less than 0.01). Neonatal serum was associated with significantly more unbound thiopental than adult serum at all levels of pH studied (P less than 0.005). Increasing the pH resulted in less free drug in both groups, but this reached statistical significance only in the adult group (P less than 0.025). Drug concentration had no effect on binding in the range examined.(ABSTRACT TRUNCATED AT 250 WORDS)

Alterations in glutathione and glutathione-related enzymes in a multidrug-resistant small cell lung cancer cell line.

H69AR is a multidrug-resistant small cell lung cancer cell line derived from a drug-sensitive cell line, H69, by selection in doxorubicin. It is cross-resistant to a wide variety of natural product-type antineoplastic agents but does not overexpress P-glycoprotein. In the present study, the levels of GSH and GSH-related enzymes in the H69AR cell line were determined and compared with those found in H69 cells. Unlike other drug-resistant cell lines, GSH levels were diminished 6-fold in H69AR cells (0.67 +/- 0.28 microgram/mg of protein), compared with H69 cells (4.23 +/- 1.17 micrograms/mg of protein) (p less than 0.01). This unusually low level of GSH may explain the pronounced collateral sensitivity of H69AR cells to buthionine sulfoximine (BSO), an inhibitor of the rate-limiting enzyme in GSH biosynthesis (ID50 of 4.4 microM BSO for H69AR cells versus ID50 of 300 microM BSO for H69 cells). BSO did not enhance doxorubicin cytotoxicity in the H69AR cell line, despite further depletion of GSH. GSH-reductase (EC 1.6.4.2) activity was elevated 2-fold in H69AR cells, compared with sensitive H69 cells (75.34 +/- 14.94 versus 38.62 +/- 5.06 nmol of NADPH/min/mg of protein) (p less than 0.05). Both selenium-dependent and -independent GSH-peroxidase (EC 1.11.1.9) activities were unchanged in the resistant H69AR cell line, compared with its parent cell line. gamma-Glutamyl transpeptidase (EC 2.3.2.2) activity was 5-fold elevated in H69AR cells, compared with H69 cells (2.50 +/- 0.44 versus 0.46 +/- 0.21 nmol of p-nitroaniline/min/mg of protein) (p less than 0.01), whereas GSH-S-transferase (EC 2.5.1.18) activity was 10-fold higher (201.98 +/- 43.62 versus 19.77 +/- 1.72 nmol of 1-chloro-2,4-dinitrobenzene/min/mg of protein in H69AR and H69 cells, respectively) (p less than 0.01). The GSH-S-transferases from both cell lines were purified by affinity chromatography and immunoblot analysis identified the GSH-S-transferases as belonging to the anionic pi class. GSH-S-transferases from the mu or alpha classes were not detectable in either cell line. In conclusion, marked differences in GSH levels and the activities of three of four GSH-related enzymes were observed between the multidrug-resistant H69AR cell line and its parent cell line. Further study is required to determine whether these changes are causally related to the development of drug resistance in this model system.

Comparison of in vitro drug responses in airways of atopic dogs with and without in vivo airway hyperresponsiveness.

For comparison with previous studies in greyhounds and in the Basenji-Greyhound dog model of asthma (BG), basenji dogs were studied under identical conditions with respect to airway responsiveness to inhaled methacholine, cutaneous responses to intradermal antigen injection, and the sensitivity of isolated trachealis muscle to methacholine and isoproterenol. The relaxant effect of isoproterenol was assessed in trachealis muscle precontracted with methacholine (ED50). The basenji dogs resembled the BG dogs in that they showed multiple positive skin tests. Further, trachealis muscle showed a markedly reduced sensitivity to methacholine (pD2 6.64 +/- 0.10) (+/- S.E.) in vitro. However, basenji dogs resembled the greyhounds in requiring high concentrations of methacholine aerosols to produce a 2-fold increase in pulmonary resistance (1.68 mg/ml +/- 1.21). Thus, there were no significant correlations between sensitivity to methacholine in vitro and airway responsiveness to methacholine in vivo; however, the reduced sensitivity to methacholine in vitro in both basenji and BG dogs may be related to the marked atopy characteristic of both groups. In vitro sensitivity to isoproterenol was correlated (r = 0.82) with the concentration of methacholine needed to elicit the test contraction, but isoproterenol sensitivity in BG dogs was significantly less (p = .0027) than that predicted by the common regression line. This deficit in beta adrenergic function in trachealis muscle unrelated to atopy may be important in the in vivo airway hyperresponsiveness of BG dogs.

Effect of calcium antagonists on the chemosensitivity of two multidrug-resistant human tumour cell lines which do not overexpress P-glycoprotein.

We have examined the ability of eight compounds to enhance adriamycin (ADM) sensitivity of two human tumour cell lines (a small cell lung cancer cell line, NCI-H69, and a fibrosarcoma cell line, HT1080) and their multidrug-resistant variants. The resistant cell lines (H69AR and HT1080/DR4) do not overexpress P-glycoprotein. Verapamil, nicardipine, perhexiline maleate, chloroquine, tamoxifen, clomiphene, prenylamine and trifluoperazine were tested alone and in combination with ADM for their cytotoxic effects. No major differences in sensitivity between the parent and resistant cell lines were noted when these agents were tested alone, except for HT1080/DR4 cells which exhibited a slight collateral sensitivity to nicardipine and H69AR cells which showed cross-resistance to chloroquine and clomiphene. When the chemosensitisers were combined with ADM no enhanced cytotoxicity of either parent cell line was observed. In HT1080/DR4 cells, verapamil showed only a modest dose-dependent chemosensitising effect while the other compounds had no effect. Verapamil and nicardipine enhanced ADM cytotoxicity in H69AR cells slightly but these effects were not dose-dependent. These results demonstrate that the reversal of drug resistance by verapamil and other calcium antagonists in a dose-dependent fashion is not an invariable property of multidrug-resistant tumour cells.

Cyclic nucleotide function in trachealis muscle of dogs with and without airway hyperresponsiveness.

We examined basal adenosine 3',5'-cyclic monophosphate (cAMP) levels, isoproterenol (ISO)-stimulated cAMP responses, basal cAMP, and guanosine 3',5'-cyclic monophosphate (cGMP) phosphodiesterase (PDE) activities and protein-kinase (PK) activities in trachealis muscle from five Basenji-greyhound (BG) and four greyhound dogs to determine whether the inverse relationship between in vivo and in vitro airway responsiveness could be due to altered cyclic nucleotide metabolism. Basal cAMP levels were not significantly different (PNS) in muscle from BG (11.6 +/- 0.53 pmol/mg protein) and greyhound dogs (10.30 +/- 1.60 pmol/mg protein). The cAMP responses to stimulation with ISO were enhanced in BG compared with greyhound dogs. The low Michaelis constant (1) for Km-cAMP PDE activity (Km = 0.63 microM) was significantly less (P less than 0.005) in BG dogs (1.54 +/- 0.28 pmol.min-1.mg protein-1) than greyhounds (11.76 +/- 2.48). Endogenously active PK activity was significantly greater (P less than 0.005) in BG (54.74 +/- 5.39 pmol.min-1.mg protein-1) than in greyhound dogs (15.50 +/0 2.20). Increases in PK activity with 5 microM cAMP added were not significantly different between BG (14.79 +/- 6.00) and greyhound dogs (7.04 +/- 2.14). Approximately 90% of both endogenous PK activity and cAMP-activated PK activity in BG and greyhound dogs was inhibited by a cAMP-dependent PK inhibitor (PKI'). These data suggest that decreased cyclic nucleotide degradation due to decreased cyclic nucleotide PDE activity with increased PK could account for the in vitro hyporesponsiveness of airway smooth muscle in BG dogs as a protective adaptive mechanism.

Large-volume ventilation results in bronchoconstriction of Basenji-Greyhound dogs.

We compared the effects of large-volume ventilation on airway responses to aerosolized histamine in anesthetized mongrel dogs with its effects in Basenji-Greyhound crossbred (B-G) dogs. Before bronchoconstriction, large inflations resulted in only small changes of dynamic compliance (Cdyn) and pulmonary resistance (RL) in both groups of dogs. After the induction of a moderate degree of bronchoconstriction with aerosolized histamine, large inflations had a more substantial effect; Cdyn increased by 7.5 +/- 2.3% (mean +/- SE; P less than 0.05), and RL decreased by 32 +/- 3.4% (P less than 0.001) in the mongrel dogs. In the B-G group, Cdyn increased by only 0.2 +/- 1.8% (NS), and RL increased by 29.3 +/- 9.2% (P less than 0.05); these changes differed significantly (P less than 0.05) from those observed in the mongrel dogs. Large-volume ventilation following the administration of indomethacin (10 mg/kg iv) and histamine increased Cdyn by 11.4 +/- 1.8% (NS vs. without indomethacin) and decreased RL by 43.9 +/- 3.4% (P less than 0.05) in the mongrel group. In the B-G group large-volume ventilation increased Cdyn by 7.6 +/- 1.7% (P less than 0.01) and decreased RL by 15.7 +/- 8.1% (P less than 0.05). Thus indomethacin enhanced the bronchodilator effects of large-volume ventilation in mongrel dogs and reversed the bronchoconstrictor effect of this maneuver on RL in B-G dogs.

Inhibition of d-tubocurarine-induced histamine release by halothane.

To determine whether anesthetics modify mediator release, the authors measured the amount of histamine released by d-tubocurarine (dTC) in human foreskin preparations in the presence of high (2.0%) and low (0.5%) halothane concentrations and nitrous oxide (10%). Freshly excised human foreskins were divided into four matched pieces. Two matched pieces were aerated with oxygen, and the other two with an oxygen-anesthetic gas mixture. One chamber of each served as a control, while the other was stimulated with 3 X 10(-5) M d-tubocurarine for 30 min. Supernatant histamine concentrations were measured by automated fluorometry. Percent histamine release was determined by dividing the experimentally released histamine concentration by the total histamine released after the tissue was sonicated and boiled. Neither halothane nor N2O alone altered spontaneous histamine release. Histamine release by d-tubocurarine was significantly reduced by 2% halothane compared to d-tubocurarine alone (2.8% +/- 0.9 vs. 13.9% +/- 3.7, mean + SEM) (P less than 0.05) in the in vitro preparation. Histamine release was reduced in the preparations pretreated with 0.5% halothane group, but this was not statistically significant (P greater than 0.05) when compared to d-tubocurarine alone. N2O (10%) did not reduce d-tubocurarine-induced histamine release. The authors conclude that halothane, in clinically used concentrations, significantly impairs histamine release from human neonatal foreskin preparations.

Comparison of drug responses in vivo and in vitro in airways of dogs with and without airway hyperresponsiveness.

Basenji-greyhound (BG) dogs demonstrate marked nonspecific airway hyperresponsiveness. To assess the possible contribution of an abnormal sensitivity of airway smooth muscle to this phenomenon, we studied the in vitro contractile responses to methacholine and histamine and the relaxant response to isoproterenol in trachealis muscle from five BG dogs with airway hyperresponsiveness in vivo and from five greyhound dogs that served as a control population. Isoproterenol responses were determined against a half-maximal methacholine contraction. Aerosol methacholine concentrations required to produce a 2-fold increase in pulmonary resistance were 0.07 +/- 0.02 (+/- S.E.) mg/ml in BG dogs and 0.67 +/- 0.26 mg/ml in greyhounds; pD2 values for methacholine-induced contraction of cervical trachealis muscle were 7.03 +/- 0.11 in BG dogs and 7.50 +/- 0.11 in greyhounds. A significant (P less than .01) negative correlation was found between methacholine sensitivity in vivo and in vitro. Aerosol concentrations of histamine required to produce a 2-fold increase in pulmonary resistance were 0.19 +/- 0.06 mg/ml in BG dogs and 1.44 +/- 0.43 mg/ml in greyhounds; pD2 values for histamine were identical in BG dogs (4.95 +/- 0.08) and greyhounds (5.05 +/- 0.19). Isoproterenol pD2 values were less in the trachealis muscle (cervical) of BG dogs (6.76 +/- 0.10) than in that of greyhounds (7.93 +/- 0.16), but this is probably a consequence of the higher concentration of methacholine needed to contract BG muscles. We conclude that the airway hyperresponsiveness of BG dogs does not reflect an increased sensitivity of airway smooth muscle per se.

Airway responses to methacholine and histamine in basenji greyhounds and other purebred dogs.

To investigate whether airway hyperresponsiveness is a general characteristic of either the basenji or the greyhound ancestry of the basenji-greyhound (BG) dog and to determine whether airway hyperresponsiveness of the BG dogs in specific for methacholine, we compared pulmonary responsiveness to increasing doses of aerosols of histamine and methacholine in 17 BG dogs, 5 unrelated purebred basenjis and 5 unrelated greyhounds. BG dogs were hyperresponsive to both methacholine and histamine compared to basenjis and greyhounds. There was a significant correlation between histamine compared to basenjis and greyhounds. There was a significant correlation between histamine and methacholine responsiveness (r = 0.70, P less than 0.0001) in the dog population. We conclude that airway hyperresponsiveness characteristic of the BG dog is not a general characteristic of the basenji or greyhound breed and occurs to histamine as well as to methacholine.

Mechanisms of intrinsic tone in bullfrog lung: relaxant effects of indomethacin, ouabain and potassium.

Bullfrog hemilungs showed minimal relaxation (9 +/- 2% of the maximal relaxant effect of theophylline, Imax) after a 16 h incubation in 10(-5) M indomethacin, indicating that prostaglandin synthesis plays little or no role in the high intrinsic tone characteristic of this preparation. A higher concentration of indomethacin (10(-4) M) produced greater relaxation (23 +/- 3% of Imax), but also markedly potentiated isoprenaline-induced relaxation. The interaction with isoprenaline was similar to that previously found for papaverine, a phosphodiesterase inhibitor. Ouabain (10(-5) and 10(-4) M) produced an initial contraction followed by marked relaxation (50% of Imax), indicating that a ouabain-sensitive mechanism is of major importance in the maintenance of intrinsic tone. Ouabain-treated hemilungs showed reversal (relaxation) of the normal contractile response to 26 mM potassium and marked impairment of the contractile response to calcium in calcium-depleted preparations. These effects suggest that ouabain-induced relaxation reflects a drug action on calcium movements. The marked relaxation (30 to 40% of Imax) produced by 26 mM potassium in ouabain-treated hemilungs is of particular interest in that it indicates a mechanism of potassium-induced relaxation distinct from stimulation of sodium-potassium ATPase.

Calcium chelators increase airway responsiveness.

To test the effect of calcium chelation on airway responsiveness to methacholine, purebred Basenji dogs were pretreated with a calcium-chelating aerosol (edetate disodium, Na2EDTA) or a placebo aerosol (saline or CaNa2-EDTA) and then challenged with methacholine bromide aerosols. The lowest dose of methacholine (0.15 mg/ml) produced no change in pulmonary resistance (RL) following pretreatment with the placebo aerosols, but RL increased (P less than 0.05) by 5.1 +/- 1.2 (SE) cmH2O X l-1 X s following pretreatment with Na2EDTA. The highest dose of methacholine (1.5 mg/ml) increased RL in all animals, but the increase was greater (P less than 0.01) following pretreatment with Na2EDTA (9.5 +/- 1.9 cm H2O X l-1 X s) than following pretreatment with a placebo aerosol (6.4 +/- 1.5 cmH2O X l-1 X s). These studies show that calcium-chelating aerosols significantly increase airway responsiveness and suggest that a localized calcium deficit may contribute to hyperresponsive airway disease.

Antigen sensitization and methacholine responses in dogs with hyperreactive airways.

Antigen sensitization was induced in six Basenji-Greyhound (BG) dogs by weekly aerosol exposure to Ascaris suum. The effects on airway responsiveness to inhaled methacholine were studied before and at least 2 wk following Ascaris sensitization. All dogs developed detectable serum levels of Ascaris-specific immunoglobulin E (IgE), and five out of six dogs developed airway responsiveness to antigen over the 4- to 6-mo period. This was accompanied by a decrease rather than an increase in airway responsiveness to inhaled methacholine. When dogs were challenged with methacholine 30 min after Ascaris antigen aerosol challenge, however, dogs reactive to Ascaris became hyperresponsive to methacholine. The magnitude of the response to antigen correlated (r = 0.85) inversely with the dose of methacholine increasing pulmonary resistance 200%. These data show that in BG dogs airway responsiveness to methacholine is increased by acute antigen exposure but that sensitization of previously unsensitized dogs does not increase nonspecific airway responsiveness.