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Biomedical subjects

R R Baker

Publications and source records attributed to R R Baker.

At least 37 records · Page 2Linked to original sources
PubMed1995

Reconstitution of immunopurified alveolar type II cell Na+ channel protein into planar lipid bilayers.

Low-amiloride-affinity (L-type) Na+ channels have been functionally and immunologically localized to alveolar type II (ATII) cells. Purified rabbit ATII epithelial cells were isolated by elastase digestion and solubilized with 3-[(3-cholamidopropyl)dimethyl-ammonio]-1-propanesulfonate. The solubilized proteins were purified by ion-exchange chromatography, followed by immunoaffinity purification over a column to which rabbit polyclonal antibodies raised against purified bovine renal Na+ channel protein were bound. The proteins eluted from the immunoaffinity column were assayed for specific binding of [3H]Br-benzamil and reconstituted into planar lipid bilayers. Sequential purification steps gave a final enrichment in specific [3H]Br-benzamil binding of > 2,000 compared with the homogenate. Single-channel currents of 25 pS were recorded from the immunopurified rabbit ATII cell protein. Addition of the catalytic subunit of protein kinase A (PKA) plus ATP to the presumed cytoplasmic side of the bilayer resulted in a significant increase in the single-channel open probability (Po), from 0.40 +/- 0.14 to 0.8 +/- 0.12, without altering single-channel conductance. The addition of amiloride or ethylisopropyl amiloride (EIPA) to the side opposite that in which PKA acts reduced Po with no change in single-channel conductance. Rabbit ATII Na+ channels in bilayers had an inhibitory constant for amiloride of 8 microM and 1 microM for EIPA. These data confirm the presence of L-type Na+ channels in adult mammalian ATII cells.

Adenosine Triphosphate↗
PubMed1994

MgATP inhibits the synthesis of 1-alkyl-2-acetyl-sn-glycero-3-phosphate by microsomal acetyltransferase of immature rabbit cerebral cortex.

The activity of 1-alkyl-sn-glycero-3-phosphate (AGP) acetyltransferase was studied using microsomal fractions isolated from cerebral cortices of 15-day-old rabbits. Fraction P3A was isolated using buffered 0.32 M sucrose containing mercaptoethanol, EDTA and NaF. This fraction had specific AGP acetyltransferase activities which were 4.9-times those of microsomal fraction P3B isolated in 0.32 M sucrose alone. This P3B activity was increased 2.4-times after a preincubation in the presence of ATP, MgCl2 and a high-speed supernatant fraction from cerebral cortex. Further, the activities of both P3A and P3B were almost completely eliminated by preincubation in the presence of alkaline phosphatase. Thus an activation of the AGP acetyltransferase by phosphorylation was indicated. While there was little inhibition of the P3A AGP acetyltransferase in the presence of added ATP, the magnesium salt form of ATP (1 mM) was severely inhibitory, bringing about 86% inhibition for P3A and 91% for P3B. The inhibitory effects of MgADP and MgAMP were smaller, and MgATP was a much more effective inhibitor than MgCTP, MgGTP and MgUTP which brought about 20-38% inhibitions of P3A activity at 1 mM concentrations. The effect of MgATP may be of particular relevance to the synthesis of platelet activating factor (PAF) following a period of ischemia in brain. Falling MgATP levels during energy failure could relieve the inhibition of AGP acetyltransferase seen in healthy cells and allow the formation of 1-alkyl-2-acetyl-sn-glycero-3-phosphate, which is the first committed intermediate in the de novo pathway of PAF synthesis.

Acetyltransferases↗
PubMed1993

The potential for platelet-activating factor synthesis in brain: properties of cholinephosphotransferase and 1-alkyl-sn-glycero-3-phosphate acetyltransferase in microsomal fractions of immature rabbit cerebral cortex.

The synthesis of platelet-activating factor (PAF) was studied in microsomal fractions of cerebral cortices of 15-day-old rabbits. These included: a total microsomal fraction P3, rough and smooth microsomes, R and S, and microsomal fraction P derived from isolated nerve cell bodies. Cholinephosphotransferase (CPT) generating PAF from alkylacetylglycerol had the highest specific activities in fractions R and P (24 and 6 times the homogenate values, based on membrane phospholipid content). This CPT activity differed from that which synthesized phosphatidylcholine as the latter was sensitive to dithiothreitol inhibition and was more readily inhibited by Triton X-100. As the CPT activity for PAF synthesis relies on the production of alkylacetylglycerol we studied the acetyltransferase which forms 1-alkyl-2-acetyl-sn-glycero-3-phosphate (AAGP). This enzyme had the highest specific activity in fraction R, followed by fractions P3 and P. There was evidence that the acetyltransferase was more active in a phosphorylated form. NaF maximized the recovery of AAGP products in the assays. The pH optimum for acetylation was in a range of 8.0-9.0. Lyso PAF did not inhibit the formation of AAGP and the rates of formation of PAF by acetylation were less than 5% of values for AAGP synthesis. During AAGP formation there was no evidence for subsequent alkylacetylglycerol formation in the absence of NaF, but a small formation of radioactive PAF could be demonstrated from AAGP under the CPT assay conditions.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetyltransferases↗
PubMed1993

Mitigation of oxidant injury to lung microvasculature by intratracheal instillation of antioxidant enzymes.

We quantitated the ability of intratracheally administered liposome-encapsulated antioxidant enzymes to reduce reactive oxygen species injury to the pulmonary microvasculature. Cationic liposomes containing 3,500 U of Cu,Zn superoxide dismutase (Cu,Zn SOD) and 3,124 U of catalase were instilled into rabbits. The animals were killed 2-72 h later and their lungs were removed and perfused with Krebs Ringer with 5% wt/vol of fat-free bovine serum albumin. The pulmonary filtration co-efficient (Kf,c) was measured before and after adding 500 microM xanthine and 5 mU/ml xanthine oxidase (XO) into the lung perfusate. Two hours after a single intratracheal instillation of liposome-entrapped Cu,Zn SOD and catalase, lung antioxidant enzyme activities were 34 and 125% higher than the corresponding control values, remained virtually unchanged for up to 8 h post-instillation, and then decreased, reaching baseline values between 24 and 72 h. Addition of xanthine and XO into the lung perfusate of un-instilled rabbits, or rabbits that received liposomes with inactivated enzymes, caused a 100% increase in Kf,c (control value: 2 +/- 0.12 ml.min-1 x cmH2O-1 per 100 g dry lung weight). On the other hand, Kf,c values of rabbits lungs instilled with liposome-encapsulated active Cu,Zn SOD and catalase and challenged with xanthine and XO 8-24 h later remained at baseline levels. Instillation of liposomes containing either enzyme was equally effective in preventing the increase in Kf,c, indicating that both superoxide anions and hydrogen peroxide were necessary for the initiation of injury. We concluded that intratracheal instillation of liposome-encapsulated antioxidant enzymes caused a transient increase of lung antioxidant enzyme levels which protects the pulmonary microvasculature from free radical-initiated injury.

Animals↗
PubMed1993

Impact of axillary lymph node dissection on the therapy of breast cancer patients.

PURPOSE: We studied a series of 283 breast cancer patients retrospectively to determine the actual benefits of axillary lymph node dissection (ALND) for these patients. PATIENTS AND METHODS: The records of 283 women with invasive breast cancer treated between 1988 and 1990 were reviewed for histologic status of the axillary lymph nodes, tumor size, DNA content, hormone-receptor values, and actual adjuvant treatments received. RESULTS: ALND was of possible therapeutic benefit for the 15% (43 of 283) of patients who had clinically positive nodes. Nodal metastases were found in 86% (37 of 43) of patients in this subgroup. ALND alone determined the indication for standard adjuvant therapy for a group of 31% (88 of 283) of patients who had favorable primary biopsy findings and clinically negative axillary nodes; ALND proved that 13% (11 of 88) of these latter patients had positive nodes. For 54% (152 of 283) of patients who had clinically negative nodes and unfavorable biopsies, ALND played no role in the decision as to whether standard adjuvant therapy was indicated. Only 5% (seven of 152) and 3% (four of 152) of these latter patients received radiation therapy and/or high-dose adjuvant chemotherapy, respectively, because of ALND. CONCLUSION: The benefits of ALND vary greatly for different groups of breast cancer patients, and controlled studies may be needed to determine whether ALND is necessary for all breast cancer patients.

Adult↗
PubMed1993

Oxidant injury to the alveolar epithelium: biochemical and pharmacologic studies.

This multifaceted study involved a combined biochemical and cellular analysis of oxidant metabolism by a lung cell at risk from injury by endogenous and environmental oxidants, the pulmonary alveolar type II epithelial cell. Within the framework of this study, a method was developed for effectively delivering antioxidant enzymes and alpha-tocopherol to the intracellular compartment of alveolar epithelial cells. Alveolar type II cells are key sources of pulmonary surfactant phospholipids and apoproteins and serve as progenitors of type I alveolar epithelium, thus playing an important role in the re-epithelialization of the lung alveolus after exposure to pulmonary oxidants. The type I and II pulmonary epithelium also play an essential collaborative role in maintaining the integrity of the air-blood barrier of the lung. Because of these critical properties of the alveolar epithelium and their recognized sensitivity to oxidant stress derived from diverse sources, such as activated inflammatory cells, hyperoxia, the environmental oxidants and nitrogen dioxide, and surgical procedures, such as cardiopulmonary bypass and lung transplantation, we endeavored to understand more about the oxidant metabolism and antioxidant pharmacology of these cells. In our experiments, we made the observation that loss of differentiated oxidant generation and antioxidant properties of type II cells occurs very rapidly in vitro. For example, we observed a 50% to 75% reduction in the specific activities of type II cell superoxide dismutase, catalase, and glutathione peroxidase, all critical scavengers of cell superoxide and hydrogen peroxide and key enzymes in the attenuation of hydroxyl radical formation. Although the differentiated characteristics of the type II cell antioxidant defenses changed in vitro, they may have become more reflective of type I alveolar epithelial cells. The type I cell is the most vulnerable for oxidant damage in the alveolus because of its large surface area and the possibility of a reduced antioxidant capacity compared to type II alveolar epithelium. In spite of this limitation, we were able to culture type II cells and study their adaptive and toxic responses to exogenously administered oxidant stress. We also observed that a significant source of self-generated oxidants in type II cells was the enzyme xanthine oxidase. Normal rates of oxidant production by this enzyme had an inhibitory effect on incorporation of biosynthetic precursors into surfactant phospholipids; these effects were eliminated by the xanthine oxidase inhibitor, allopurinol.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1992

The hydrolysis of natural phosphatidylethanolamines by phospholipase A2 from rat serum: a degree of selectivity is shown for docosahexaenoate release.

The selectivity of phospholipase A2 from serum was evaluated using radioassays and mass analyses of fatty acids liberated from phosphatidylcholine and phosphatidylethanolamine. These natural phospholipid substrates were labelled at the sn-2 position with radioactive oleate, linoleate and arachidonate. The rates of release of fatty acids were compared with their abundance at the sn-2 position of these phospholipid substrates. While there was little or no selectivity in the liberation of these fatty acids from phosphatidylcholine, there was some evidence for a preferential release of arachidonate with respect to linoleate from phosphatidylethanolamine. Mass analyses of free fatty acid products revealed that docosahexaenoate was consistently liberated at levels that exceeded its abundance at the sn-2 position of phosphatidylethanolamine. Three different, natural phosphatidylethanolamines with varying levels of docosahexaenoate showed a 1.2-1.8-fold enrichment of this polyunsaturate in the free fatty acid products compared with its abundance at the sn-2 position. This preference could also be shown when phosphatidylethanolamine was mixed with synthetic phosphatidylcholine as co-sonicated substrates. This preferential release of docosahexaenoate by serum phospholipase A2 is of considerable significance in the nervous system which is enriched in this polyunsaturate. The potential competition between liberated docosahexaenoate and arachidonate may be of fundamental importance in the response of brain to hemorrhage.

Animals↗
PubMed1992

Quantitation of alveolar distribution of liposome-entrapped antioxidant enzymes.

Liposome-encapsulated Cu,Zn superoxide dismutase (Cu,Zn SOD) and catalase (CAT) were instilled intratracheally in rabbits, and the temporal and spatial distribution of Cu,Zn SOD and CAT within the lung was assessed at the organ and cellular levels. Specific activities of Cu,Zn SOD and CAT were increased in both lung homogenates and isolated alveolar type II pneumocytes. Peak Cu,Zn SOD activities in lung homogenates and alveolar type II cells were observed 4 h after liposome instillation and returned to control levels by 24 h, whereas CAT activities remained significantly above controls. There were no significant differences in liposome distribution or antioxidant enzyme uptake among lung lobes. The distribution of fluorescently labeled Cu,Zn SOD and CAT was assessed with the use of epifluorescence microscopy and digital image processing to determine patterns of cellular incorporation of liposome-entrapped Cu,Zn SOD and CAT within the lung. Although the mean fluorescence intensity of alveoli from rabbits instilled with liposomes containing labeled Cu,Zn SOD and CAT was greater than autofluorescence observed with either no liposome or empty liposome instillation, fluorescence intensity varied between adjacent alveoli. Both fluorescently labeled Cu,Zn SOD and CAT were located cytosolically, and uptake was not limited to alveolar type II pneumocytes. These results demonstrate that a single intratracheal instillation of liposomes can effect increases in Cu,Zn SOD and CAT activities in distal lung cells, including alveolar type I and type II cells and macrophages.

Animals↗
PubMed1992

The management of breast cancer with immediate or delayed reconstruction.

In patients with stage I and stage II disease who are candidates for a mastectomy, either immediate or delayed reconstruction can be accomplished with remarkably good cosmetic results. Age is not a specific contraindication to reconstruction, but, in my experience, the majority of women over 55 years of age are not interested. Mastectomy or modified radical mastectomy should be performed in these patients using a technique that facilitates delayed reconstruction. The patient may or may not elect to have the reconstruction eventually. Immediate or delayed reconstruction is appealing to a significant number of women under 55 years of age. They should see a plastic surgeon in consultation and the advantages and disadvantages of a subpectoral silicone implant vs. a myocutaneous flap (usually a transverse rectus abdominis musculocutaneous flap) should be discussed in detail. The patient's acceptance of the cosmetic result is better if she has an opportunity preoperatively to review photographs of reconstructed breasts and to discuss the results with women who have undergone a similar procedure. The timing of the reconstruction is a matter of clinical judgment; no prospective clinical trials have been performed. Immediate reconstruction is feasible if the skin flaps will accept the slight tension produced by either a subpectoral implant or a transposed muscle flap. A subpectoral silicone implant produces a satisfactory cosmetic result in the majority of patients, particularly if both breasts are supported by a properly fitting brassiere. In long-term follow-up, infection, contracture, and displacement requiring removal and reinsertion occur in approximately 15% of patients. Reconstruction by a myocutaneous flap is a technically more difficult operation. The operating time is prolonged significantly and blood loss is increased. In my opinion, a myocutaneous flap probably is not worth the risk of a heterologous transfusion. The long-term cosmetic results, however, are considerably better in comparison to a subpectoral silicone implant. The reconstructed breast is much more mobile, making it a more realistic replacement of the original breast. In most cases, it is better to perform this type of reconstruction with a myocutaneous flap as a delayed procedure. In experienced hands, however, immediate reconstruction, with the transverse rectus abdominis musculocutaneous flap harvested while the modified radical mastectomy is being performed, is perfectly feasible. Nipple/areolar reconstruction is available. However, the cosmetic results are only fair to good, and the patient should be aware of this before undergoing the procedure. The problem of symmetry between the breasts requires a good deal of judgment.(ABSTRACT TRUNCATED AT 400 WORDS)

Adult↗
PubMed1991

Mechanisms of H2O2-mediated injury to type II cell surfactant metabolism and protection with PEG-catalase.

Alterations in type II pneumocyte function, including surfactant biosynthesis, may play a significant role in the development and pathophysiology of oxidant-induced lung injury. The results of this study showed that type II cells exposed to 50-300 microM H2O2 demonstrated a dose-dependent decrease in phosphatidylcholine (PC) synthesis with only minimal changes in cell viability. The activities of the choline-phosphate cytidyltransferase and cholinephosphotransferase, specific enzymes of PC synthesis, were not significantly decreased by the exposure. However, the activity of glycerol-3-phosphate acyltransferase, a sulfhydryl-dependent enzyme involved in an early stage of phospholipid synthesis, was decreased by the exposures in a manner that was similar to that seen for PC synthesis. Further studies showed that incubation of type II cells with polyethylene glycol-conjugated catalase for 1 h resulted in an increase in the cell-associated catalase activity (53 +/- 5 vs. 6.7 +/- 1.5 units/mg protein for controls). Confocal microscopy analysis showed that a significant portion of this activity was located intracellularly. More importantly, these cells were protected from changes in PC synthesis rates when subsequently incubated with 300 microM H2O2. These results indicate that the deleterious effects of H2O2 on type II cell surfactant synthesis may be pharmacologically modified in vitro, a concept that may have utility with regard to the modulation of in vivo lung injuries.

Animals↗
PubMed1990

Characterization of antioxidant activities of pulmonary surfactant mixtures.

Instillation of intratracheal surfactant is known to limit the morbidity and mortality of patients and animals with oxidant-induced lung injury. In this study we quantified the antioxidant properties of natural lung surfactant (NLS), consisting of 90% lipid and 10% protein, and of calf lung surfactant extract (CLSE) consisting of 99% lipid and 1% protein. NLS, but not CLSE, contained significant amounts of superoxide dismutase (SOD) and catalase activities (7 U SOD/mumol phospholipid (PL) and 1 U catalase/mumol PL). More than 90% of the SOD activity was abolished by 1 mM KCN, suggesting that this was the CuZn form of the enzyme. In addition, NLS significantly reduced extracellular H2O2 without losing its ability to reach minimum surface tensions below 1 dyn/cm upon dynamic compression. The NLS scavenging of H2O2 could not be accounted for by albumin. The presence of catalase and SOD activities in NLS was also verified by activity stains of proteins separated by native polyacrylamide gel electrophoresis. Intratracheal instillation of 7 ml of NLS (308 mumol PL) into rabbits significantly increased SOD content in type II cells isolated 12 h later. It is concluded that, in addition to promoting alveolar stability, instillation of pulmonary surfactant may offer significant protection to the alveolar epithelium by scavenging extracellularly generated partially reduced oxygen species and by enhancing intracellular antioxidant enzyme content.

Animals↗
PubMed1990

Phosphatidylinositol synthetase activities in neuronal nuclei and microsomal fractions isolated from immature rabbit cerebral cortex.

The synthesis of phosphatidylinositol was studied using a nuclear fraction N1, a microsomal fraction P3, rough (R) and smooth (S) microsomal fractions and a microsomal fraction P derived from isolated nerve cell bodies. Each fraction was prepared using cerebral cortices of 15-day-old rabbits. In assays using CDP-diacylglycerol (prepared from egg phosphatidylcholine) and myo[3H]inositol at pH 7.4, fraction N1 had the highest maximal specific rates of phosphatidylinositol synthetase (EC 2.7.8.11) (expressed per mumol phospholipid in the fraction). However the three microsomal fractions achieved maximal specific activities at liponucleotide concentrations close to 50 microM, while fraction N1 required 200 microM concentrations. In certain cases (25-120 microM CDP-diacylglycerol, and at higher pH values) fraction R had specific activities which equalled or surpassed those of N1. However, with respect to inositol, fraction N1 had a distinctly lower Km than was shown for fractions R or P3. Each of the microsomal fractions and N1 required Mg2+ for the reaction, but for N1, maximal rates could be sustained at 0.1 mM, while for the microsomal fractions the optimal Mg2+ concentration was 1 mM. For each fraction Mn2+ could not replace Mg2+ in the reaction and Mn2+ was inhibitory. The optimal pH for the reaction was between 8.0 and 9.0. Phosphatidylinositol synthetase could also be shown using fraction N1 enriched in endogenous CDP-diacylglycerol. The relatively high specific activities of fraction N1, and the differences found between N1 and the microsomal fractions, for optimal CDP-diacylglycerol and Mg2+ concentrations and for Km values for inositol, support the existence of a neuronal nuclear phosphatidylinositol synthetase.

Animals↗
PubMed1990

The eicosanoids: a historical overview.

Eicosanoids are biologically active compounds derived from 20 carbon unsaturated fatty acids, among which arachidonic acid is a substrate of particular importance. The history of the eicosanoids dates back to the thirties, when new biologically active compounds were found in human seminal plasma. These "prostaglandins" were purified, and their structures and mechanisms of biosynthesis were elucidated in the early sixties. Other eicosanoids, including thromboxane A2, a potent platelet aggregating agent, and prostacyclin, an antagonist to thromboxane A2, were discovered in the seventies. The inhibitory actions of acetylsalicylic acid on eicosanoid synthesis were also uncovered at this time. In 1979, a new metabolic sequence leading to the synthesis of a new group of eicosanoids, called leukotrienes, was reported. The leukotrienes have several biological activities, including the mediation of bronchoconstriction in allergic response. The eicosanoids comprise a diverse group of biologically active compounds; many of these arise from arachidonic acid, and are associated with injury, allergic responses, and platelet aggregation.

Eicosanoids↗
PubMed1990

Unusual lesions and their management.

Treatment of a breast lesion should never be undertaken without a precise histologic diagnosis, which means careful examination of multiple sections of tissue embedded in paraffin and appropriately stained. The author described the clinical and histologic features of lobular and medullary cancers; cystosarcoma phylloides; carcinosarcoma; Paget's disease; adenoid cystic carcinoma; mucinous, tubular, and papillary cancers; and other unusual lesions that can originate elsewhere in the body. For each, treatment suggestions are offered.

Adenocarcinoma↗
PubMed1990

Fatty acid release in incubations of serum with synaptosome and myelin subfractions of brain.

To study lipid breakdown in brain membranes following hemorrhage, synaptosome and myelin fractions isolated from rat brain were incubated with rat serum. After 3 h in vitro at 37 degrees C, 0.43 and 0.26 mumol of fatty acid were released in incubations containing synaptosomes (1.37 mumols phospholipid) or myelin (1.23 mumols phospholipid), respectively, in the presence of 0.25 mL serum. Less than 0.05 mumol of fatty acid was liberated in incubations containing only serum, synaptosomes, or myelin. For synaptosomes and serum, docosahexaenoate was the principal fatty acid released (28 mol% of total) after 3 h of incubation. This fatty acid and arachidonate made up 43 mol% of the liberated fatty acid. The presence of free docosahexaenoate was of interest, as this fatty acid is particularly enriched in phosphatidylserine and phosphatidylethanolamine, phospholipids found in the cytoplasmic half of the synaptosomal plasma membrane and in interior synaptosomal membranes. In incubations of serum and myelin, oleate was the major free fatty acid produced in 30 min to 3 h of incubation (29-35 mol% of total). After 3 h, docosahexaenoate contributed 20 mol% to the total. The release of fatty acids from the membranes may be mediated by serum phospholipase(s) or possibly by activated endogenous lipolytic activities.

Animals↗
PubMed1990

The acylation of 1-acyl-sn-glycero-3-phosphate by neuronal nuclei and microsomal fractions of immature rabbit cerebral cortex.

The acylation of 1-acyl-sn-glycero-3-phosphate to form phosphatidic acid was studied using a neuronal nuclear fraction N1 and microsomal fractions P3, R (rough), S (smooth), and P (neuronal microsomes from nerve cell bodies) isolated from cerebral cortices of 15-day-old rabbits. The assays contained this lysophospholipid, ATP, CoA, MgCl2, NaF, dithiothreitol, and radioactive palmitate, oleate, or arachidonate. Of the subfractions, N1 and R had the highest specific activities (expressed per micromole phospholipid in the fraction). The rates with oleate were two to four times the values seen for phosphatidic acid formation from sn-[3H]glycero-3-phosphate and oleoyl-CoA. Using oleate or palmitate, fraction R had superior specific rates to N1 at low lysophosphatidic acid concentrations. With increasing lysophospholipid concentrations the specific rates of N1 and R came closer together and maintained at least a twofold superiority over fraction P. Fraction S had the lowest specific rates of phosphatidic acid formation. Fractions N1, R, and P showed a preference for palmitate and oleate over arachidonate, particularly at low concentrations of lysophosphatidic acid. For N1 and R, the preference was also more marked at higher concentrations of fatty acid. Thus a selectivity for saturated and monounsaturated fatty acids was shown in the formation of phosphatidic acid, as was a concentration of acylating activity in the neuronal nucleus and the rough endoplasmic reticulum.

Acylation↗
PubMed1990

Endogenous xanthine oxidase-derived O2 metabolites inhibit surfactant metabolism.

The ability of xanthine oxidase (XO)-derived, partially reduced O2 species (PROS) to inhibit surfactant production was examined in freshly isolated alveolar type II (ATII) pneumocytes from New Zealand White rabbits. [Methyl-3H]choline chloride and [1-14C]palmitate incorporation into phosphatidylcholine (PC) decreased in a dose-dependent manner, whereas peak media hydrogen peroxide (H2O2) concentration increased, when 1, 5, or 10 mU/ml XO were added to cell suspensions containing 500 microM xanthine. Addition of 100 microM allopurinol inhibited H2O2 production and abolished the decrease in choline and palmitate incorporation into PC. ATII cells incubated with 500 microM xanthine alone incorporated choline and palmitate at 90 and 80% of control levels, respectively. However, 100 microM allopurinol restored precursor incorporation to control values. To identify a possible intracellular source of PROS, ATII cell xanthine dehydrogenase (XDH) and XO activities were measured. Both total activity (XDH + XO; 45 +/- 7 microU/mg protein) and the percentage activity in the oxidase form (%XO; 30 +/- 4%) remained unchanged in ATII cells incubated in media only (control) for 2 h. In contrast, incubation of ATII cells with 500 microM xanthine resulted in a 50% loss of XDH + XO activity and a 21% increase in %XO within 10 min. After 2 h there was no measurable XDH + XO activity in xanthine-treated cells. Total XDH + XO activity in cells incubated with 500 microM xanthine and 100 microM allopurinol was less than 6% of control values throughout the incubation.(ABSTRACT TRUNCATED AT 250 WORDS)

Allopurinol↗