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P W Cheng

Publications and source records attributed to P W Cheng.

At least 19 recordsLinked to original sources
PubMed1995

Alteration of sulfation of glycoconjugates, but not sulfate transport and intracellular inorganic sulfate content in cystic fibrosis airway epithelial cells.

The secreted and cell surface high molecular weight glyco-conjugates (HMG) generated by primary cultures of airway epithelial cells from cystic fibrosis (CF) patients are oversulfated. To determine whether this abnormality is maintained in transformed CF airway epithelial cells and whether differences in transport or intracellular accumulation of sulfate can explain this alteration, we assessed sulfate metabolism in paired CF and normal cell lines as well as primary cultures of CF and normal cells. Both 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid-inhibitable and -resistant [35S]sulfate efflux and influx were identical for each pair of CF and normal cell lines. Furthermore, cell content of inorganic sulfate was not significantly different in CF and normal cells. However, compared with primary CF cells that oversulfate HMG transformed CF cells oversulfated cell surface HMG but not HMG released into culture medium. Our results suggest that plasma membrane sulfate transport is not altered in CF airway epithelial cells and the abnormal sulfation of HMG may be due to perturbation in intracellular sulfate activation or transfer of activated sulfate to HMG. The relationship of this abnormality to CF transmembrane conductance regulator mutations remains to be determined.

Biological Transport↗
PubMed1994

Lectin conjugate-directed gene transfer to airway epithelial cells.

Many cell surface receptors contain covalently linked carbohydrates recognizable by lectins. This study examined the feasibility of using lectins as gene delivery vectors. Polylysine (PL) and histone (His) conjugates of concanavalin A (Con A), Maackia amurensis agglutinin (MAA), Sambucus nigra agglutinin (SNA), and wheat germ agglutinin (WGA), were employed to deliver pCMVlacZ to human airway epithelial cells (CFT1). As compared to the control (= 1.0), the beta-galactosidase activities expressed in CFT1 cells transfected with lectin-PL/His-DNA containing 5 micrograms/ml conjugate and 10 micrograms/ml DNA were: Con A-His, 7.7; SNA-His, 3.5; Con A-PL, 2.4; WGA-PL/His, 1.3; others, 1.0. The efficiency of gene transfer directed by Con A-His was dose dependent between 2.5 and 10 micrograms/ml of the conjugate. The maximal beta-galactosidase activity expressed relative to the control was 42 obtained using 10 micrograms/ml conjugate. These results demonstrate that lectin-PL/His conjugates can be used as vectors to transfer genes to airway epithelial cells.

Binding Sites↗
PubMed1994

Production and characterization of monoclonal antibodies against guinea pig tracheal mucins.

Thirty-five hybridomas that secrete mouse monoclonal antibodies (MAb) against guinea pig (G.P.) tracheal mucins were established. The MAbs were characterized immunologically, biochemically, and immunohistochemically at both light and electron microscopic levels. Isotyping of the MAbs revealed 14 to be IgM, 13 IgG1, 3 IgG2, and 5 IgG3. The MAbs demonstrated various patterns of binding in immunoblots against mucins derived from G.P. tracheal explants. This suggested the presence of "subpopulations" of G.P. tracheal mucins with specific MAbs binding to different epitopes on the mucin molecules. Periodate oxidation indicated that 33 of the 35 MAbs recognized carbohydrate epitopes on the mucin molecules. Ten of the MAbs also reacted with both bovine and ferret tracheal mucins, while 7 and 6 MAbs bound only to bovine and ferret tracheal mucins, respectively. The generated MAbs should be useful for immunomeasurement of mucin secretion in vivo (e.g., in bronchoalveolar or airway lavage fluid) and in vitro (e.g., cell and organ cultures) from cells of guinea pig and (with certain MAbs) bovine and ferret origin.

Animals↗
PubMed1994

The role of airway mucus in pulmonary toxicology.

Airway mucus is a complex airway secretion whose primary function as part of the mucociliary transport mechanism is to to serve as renewable and transportable barrier against inhaled particulates and toxic agents. The rheologic properties necessary for this function are imparted by glycoproteins, or mucins. Some respiratory disease states, e.g., asthma, cystic fibrosis, and bronchitis, are characterized by quantitative and qualitative changes in mucus biosynthesis that contribute to pulmonary pathology. Similar alterations in various aspects of mucin biochemistry and biophysics, leading to mucus hypersecretion and altered mucus rheology, result from inhalation of certain air pollutants, such as ozone, sulfur dioxide, nitrogen dioxide, and cigarette smoke. The consequences of these pollutant-induced alterations in mucus biology are discussed in the context of pulmonary pathophysiology and toxicology.

Air Pollutants↗
PubMed1993

Evoking the permission schema: the impact of explicit negation and a violation-checking context.

Cheng and Holyoak (1985) proposed that realistic reasoning in deontic contexts is based on pragmatic schemas such as those for assessing compliance with or violation of permission and obligation rules, and that the evocation of these schemas can facilitate performance in Wason's (1966) selection task. The inferential rules in such schemas are intermediate in generality between the content-independent rules proposed by logicians and specific cases stored in memory. In one test of their theory, Cheng and Holyoak demonstrated that facilitation could be obtained even for an abstract permission rule that is devoid of concrete thematic content. Jackson and Griggs (1990) argued on the basis of several experiments that such facilitation is not due to evocation of a permission schema, but, rather, results from a combination of presentation factors: the presence of explicit negatives in the statement of cases and the presence of a violation-checking context. Their conclusion calls into question both the generality of content effects in reasoning and the explanation of these effects. We note that Jackson and Griggs did not test whether the same combination of presentation factors would produce facilitation for an arbitrary rule that does not involve deontic concepts, as their proposal would predict. The present study tested this prediction. Moreover, we extended Jackson and Griggs' comparisons between performance with an abstract permission rule versus an arbitrary rule, introducing clarifications in the statement of each. No facilitation was observed for an arbitrary rule even when explicit negatives and a violation-checking context were used, whereas strong facilitation was found for the abstract permission rule under the same conditions. Performance on the arbitrary rule was not improved even when the instructions indicated that the rule was conditional rather than biconditional. In contrast, a small but reliable degree of facilitation was obtained for the abstract permission rule, with violation-checking content even in the absence of explicit negatives. The theory of pragmatic reasoning schemas can account for both the present findings and those reported by Jackson and Griggs.

Adolescent↗
PubMed1993

Sulfate concentrations and transport in human bronchial epithelial cells.

Inorganic sulfate concentrations in the cytoplasm of human bronchial epithelial cells exceeded levels in the bathing medium under all circumstances tested. Cell sulfate concentrations were directly related to medium sulfate concentrations and inversely related to medium chloride concentrations. In physiological media there was a sulfate compartment of approximately 0.3 mM that exchanged very slowly with extracellular sulfate. In media lacking chloride, sulfate was accumulated by the cells to a level as high as 2 mM. Sulfate uptake was markedly inhibited by external chloride and by stilbene sulfonic acid derivatives but was not affected by sodium in the medium. Efflux of 35SO4(2-) was stimulated by both chloride and sulfate in the bathing medium but inhibited by stilbenes. The following compounds had no effect on sulfate movements: phorbol esters, adenosine 3',5'-cyclic monophosphate derivatives, and okadaic acid. Changes in medium tonicity were likewise without effect. Our results suggest that human bronchial epithelial cells maintain a steady-state disequilibrium for inorganic sulfate. Furthermore, sulfate appears to exist in at least two compartments in the cells: one that is slowly exchangeable with sulfate in the medium and another exchangeable compartment that is of negligible size in physiological media but that becomes very large in media lacking chloride. Sulfate is transported by an anion exchanger of broad specificity that is not influenced by substances known to modulate chloride channels.

Biological Transport↗
PubMed1993

Increased adherence of Staphylococcus aureus from cystic fibrosis lungs to airway epithelial cells.

Airway colonization by Staphylococcus aureus is a frequent feature of cystic fibrosis (CF). To assess the pathogenesis of selective colonization with this organism, we compared the capacity of S. aureus isolated from the respiratory tract of CF and non-CF patients to adhere to epithelial cells from the upper and lower airways of CF and control subjects. Bacterial adherence to bronchial epithelial cell lines was significantly greater for CF than for non-CF isolates (p < 0.001). Of 17 CF S. aureus isolates 12 adhered at a level > 1 bacterium per cell; this was true for only 1 of 14 non-CF isolates. CF S. aureus isolates also bound more avidly than non-CF isolates to ciliated (p < 0.05) and squamous nasal cells (p < 0.02) and buccal epithelial cells (p < 0.005) freshly harvested by scraping. Each S. aureus isolate bound with equal avidity to epithelial cells from CF patients and healthy individuals. Adherence was not related to sex, age, severity of pulmonary disease, presence of other microorganisms in the airways, or genotype of the CF hosts. Binding of S. aureus was blocked by proteinase treatment of organisms, suggesting that adherence is mediated by one or more peptide adhesins. We propose that the high prevalence of adherent S. aureus is due either to selection of adherent strains by CF airways or to induction of an adherent phenotype by factors residing at the CF airways surface.

Adolescent↗
PubMed1992

Covariation in natural causal induction.

The covariation component of everyday causal inference has been depicted, in both cognitive and social psychology as well as in philosophy, as heterogeneous and prone to biases. The models and biases discussed in these domains are analyzed with respect to focal sets: contextually determined sets of events over which covariation is computed. Moreover, these models are compared to our probabilistic contrast model, which specifies causes as first and higher order contrasts computed over events in a focal set. Contrary to the previous depiction of covariation computation, the present assessment indicates that a single normative mechanism--the computation of probabilistic contrasts--underlies this essential component of natural causal induction both in everyday and in scientific situations.

Cognition↗
PubMed1991

Mucin biosynthesis: purification and characterization of a mucin beta 6N-acetylglucosaminyltransferase.

We have purified, to apparent homogeneity, a mucin beta 6N-acetylglucosaminyltransferase (beta 6GlcNAc transferase) from bovine tracheal epithelium. Golgi membranes were isolated from a 0.25 M sucrose homogenate of epithelial scrapings by discontinuous sucrose gradient centrifugation. The Golgi membranes were solubilized with 1% Triton X-100 in the presence of 1 mM Gal beta 1-3GalNAc alpha benzyl (Bzl) to stabilize the beta 6GlcNAc transferase. The solubilized enzyme was bound to a UDP-hexanolamine-Actigel-ALD Superflow affinity column equilibrated with 1 mM Gal beta 1-3GalNAc alpha Bzl and 5 mM Mn2+. Elution of the enzyme with 0.5 mM UDP-GlcNAc resulted in a 133,800-fold purification with a 1.3% yield and a specific activity of 70 mumol/min/mg protein. Radioiodination of the purified enzyme followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography revealed a single band at 69,000 Da. Kinetic analyses of the beta 6GlcNAc transferase-catalyzed reaction showed an ordered sequential mechanism in which UDP-GlcNAc binds to the enzyme first and UDP is released last. The Km values for UDP-GlcNAc and Gal beta 1-3GalNAc alpha Bzl were 0.36 and 0.14 mM, respectively. Acceptor competition studies showed that the purified beta 6GlcNAc transferase can use core 1 and core 3 mucin oligosaccharides as well as GlcNAc beta 1-3Gal beta R as acceptor substrates. Proton NMR analyses of the three products demonstrated that GlcNAc was added in a beta 1-6 linkage to the penultimate GalNAc or Gal, suggesting that this enzyme is capable of synthesizing all beta 6GlcNAc structures found in mucin-type oligosaccharides.

Animals↗
PubMed1991

Mucin-like glycoprotein secreted by cultured hamster tracheal epithelial cells. Biochemical and immunological characterization.

We isolated mucin-like glycoproteins from the conditioned medium of primary hamster tracheal epithelial (HTE) cell culture and characterized them biochemically and immunologically. These glycoproteins were purified on Sepharose CL-4B after Streptomyces hyaluronidase treatment and then by CsCl-density-gradient centrifugation in the presence of 4 M-guanidinium chloride. The purified glycoproteins were resistant to digestion by chondroitin AC lyase, heparinase, heparitinase and endo-N-acetylglucosaminidases A, D and H, but susceptible to endo-beta-galactosidase and keratanase. SDS/PAGE demonstrated no contamination by low-molecular-mass proteins. The purified glycoproteins showed a peak buoyant density of 1.56 g/ml in CsCl-density-gradient centrifugation, and contained 10% peptide and 90% carbohydrate by weight. Carbohydrates in these glycoproteins contained N-acetylglucosamine, N-acetylgalactosamine, galactose, fucose, sialic acid and a trace amount of mannose, but no uronic acid. Serine and threonine together accounted for 27% of the total amino acid residues. In addition, the mucin-like glycoproteins exhibited blood-group A and B activities, and very strong inhibitory activity for influenza A virus haemagglutination. With the use of the purified glycoprotein as an antigen, six monoclonal antibodies that stained mucus granules in hamster tracheal epithelium were obtained. We characterized the antibody produced by one of the clones, HM D46. We conclude that HTE cells cultured in the serum-free medium secrete a glycoprotein with physicochemical properties similar to those known in various airways mucins.

Amino Acids↗
PubMed1991

Enhanced host cell reactivation of damaged plasmid DNA in HeLa cells resistant to cis-diamminedichloroplatinum(II).

Human HeLa cells resistant to cisplatin were established by stepwise selection. The selected cells showed a 15- to 20-fold cisplatin resistance (CPR) at the dose level resulting in 50% inhibition. These cells were cross-resistant to mitomycin C, melphalan, and ethyl methanesulfonate but not to Adriamycin, colchicine, or vinblastine. The expression of cisplatin-damaged plasmid DNA carrying the bacterial chloramphenicol acetyltransferase (CAT) gene after its transfection into CPR cells was enhanced by approximately 3-fold. This did not correlate with the degree of CPR. However, the development of the CPR phenotype paralleled the enhanced CAT activity. The addition of aphidicolin (an inhibitor of DNA alpha-polymerase) to CPR cells effectively diminished the enhanced CAT activity and CPR. These studies have identified an enhanced host cell reactivation of the damaged plasmid in the acquisition of CPR, suggesting that DNA repair is a potential mechanism for the development of CPR phenotype in human cells.

Antineoplastic Agents↗
PubMed1991

Causes versus enabling conditions.

People distinguish between a cause (e.g., a malfunctioning component in an airplane causing it to crash) and a condition (e.g., gravity) that merely enables the cause to yield its effect. This distinction cannot be explained by accounts of reasoning formulated purely in terms of necessity and sufficiency, because causes and enabling conditions hold the same logical relationship to the effect in those terms. Proposals to account for this apparent deviation from accounts based on necessity and sufficiency may be classified into three types. One approach explains the distinction in terms of an inferential rule based on the normality of the potential causal factors. Another approach explains the distinction in terms of the conversational principle of being informative to the inquirer given assumptions about his or her state of knowledge. The present paper evaluates variants of these two approaches, and presents our probabilistic contrast model, which takes a third approach. This approach explains the distinction between causes and enabling conditions by the covariation between potential causes and the effect in question over a focal set--a set of events implied by the context. Covariation is defined probabilistically, with necessity and sufficiency as extreme cases of the components defining contrasts. We report two experiments testing our model against variants of the normality and conversational views.

Adult↗
PubMed1990

Increased mdr gene expression and decreased drug accumulation in a human colonic cancer cell line resistant to hydrophobic drug.

An adriamycin-resistant human colonic cancer cell line was characterized. This clone exhibits the classical multidrug resistance (MDR) phenotype, being cross-resistant to hydrophobic drugs such as colchicine, and vinblastine. In contrast, this clone shows a normal response to DNA-damaging agents. The appearance of MDR in these cells was linked to a decreased accumulation of the drug [3H]colchicine as compared to the drug-sensitive cells. This MDR line expressed 80-100 fold increased levels of the specific 4.5-kb mdr mRNA, and a gene amplification. Our results indicate that MDR in human colonic cancer cells can result from increased expression of at least one member of the mdr gene family.

Antineoplastic Agents↗
PubMed1990

Enzymatic synthesis of UDP-GlcN by a two step hollow fiber enzyme reactor system.

UDP-GlcN was synthesized from GlcN and UTP by a two step hollow fiber enzyme reactor method. In step 1, GlcN was converted to GlcN 6-P and then to GlcN 1-P by hexokinase and phosphoglucomutase, respectively, and UTP was used as the phosphate donor. In step 2, GlcN 1-P was converted to UDP-GlcN by UDP glucose pyrophosphorylase. All the enzymes required for the synthesis of UDP-GlcN were enclosed in hollow fiber bundles which allow for the free diffusion of substrates and products across the membranes to and from the enzymes, allow for the reutilization of the enzymes, and simplify the isolation of the product, UDP-GlcN. We show that both UTP and GlcN 6-P are inhibitors of the yeast UDPG pyrophosphorylase and therefore their concentrations must be regulated to obtain maximum yields of UDP-GlcN. The UDP-GlcN produced can be N-acetylated with [14C]acetic anhydride to produce UDP-[14C]GlcNAc. This method can also be used to synthesize [32P]UDP-GlcN and [32P]UDP-GlcNAc from [alpha-32P]UTP and GlcN 1-P.

Acetylglucosamine↗
PubMed1990

A probabilistic contrast model of causal induction.

Deviations from the predictions of covariational models of causal attribution have often been reported in the literature. These include a bias against using consensus information, a bias toward attributing effects to a person, and a tendency to make a variety of unpredicted conjunctive attributions. It is contended that these deviations, rather than representing irrational biases, could be due to (a) unspecified information over which causal inferences are computed and (b) the questionable normativeness of the models against which these deviations have been measured. A probabilistic extension of Kelley's analysis-of-variance analogy is proposed. An experiment was performed to assess the above biases and evaluate the proposed model against competing ones. The results indicate that the inference process is unbiased.

Adult↗
PubMed1990

Characterization of guinea pig tracheal epithelial cells maintained in biphasic organotypic culture: cellular composition and biochemical analysis of released glycoconjugates.

An air-liquid interface (biphasic) primary culture system in which guinea pig tracheal epithelial cells maintain morphologic characteristics of differentiated epithelium has been developed in this laboratory. In this report, we compared quantitatively cell populations of 8-day cultures to those of epithelial mucosa in intact trachea. In addition, high molecular weight glycoconjugates released by the cultured cells were isolated and characterized. Quantitative morphometric analysis revealed similar volume densities of ciliated, secretory, basal, and "other" cells in cultures and in intact tracheal surface epithelium, although the cultures tended to have smaller cells and contained fewer basal cells. High molecular weight glycoconjugates released apically by cell cultures and excluded from Sepharose CL-4B columns contained approximately 5% hyaluronic acid but undetectable amounts of other proteoglycans, such as chondroitin sulfate, heparan sulfate, and dermatan sulfate. The hyaluronidase-resistant glycoconjugates exhibited a peak buoyant density at 1.49 g/ml on cesium chloride density gradient centrifugation and were shown to contain mucin-type carbohydrate to peptide linkages (i.e., GalNAc to ser/thr) and an amino acid composition typical of respiratory mucins. The results indicate that this organotypic cell culture system mimics quite closely morphology of mucosal epithelium in intact airways and that the cells release high molecular weight glycoconjugates with biochemical properties of mucin-type glycoproteins. Thus, this in vitro system appears well-suited for studies of mucin secretion and other functions of respiratory epithelial cells.

Amino Acids↗
PubMed1989

Developmental changes of ferret tracheal mucin composition and biosynthesis.

We characterized the chemical composition of mucins secreted by ferret tracheal explants and the activities of key mucin glycosyltransferases in ferret tracheal epithelium during a period of rapid postnatal maturation of the mucin-secreting structures. Ferret tracheal explants secrete three major groups of high molecular weight glycoconjugates: (1) those susceptible to bovine testicular hyaluronidase; (2) those resistant to hyaluronidase and exhibiting high density (p greater than or equal to 1.60 g/mL); and (3) those resistant to hyaluronidase and exhibiting low density (1.45 less than or equal to p less than 1.60 g/mL). The hyaluronidase-resistant, low-density glycoconjugates have typical mucin properties and constitute 36% of total glycoconjugates released in newborns but only 8% in adult ferrets. Mucin secretory rate per unit surface area of trachea progressively decreases with age. Mucin amino acid and total carbohydrate contents do not vary; however, the sialic acid content increases, and fucose content as well as blood group A activity of the mucins decreases with age. Four glycosyltransferases involved in mucin biosynthesis [Gal beta 3GalNAc:(GlcNAc-GalNAc)beta 6 N-acetylglucosaminyl-, GalNAc:beta 3 galactosyl-, Gal:alpha 2 fucosyl-, and GalNAc alpha 2----6 neuraminyltransferase] are present in tracheal epithelium of ferrets at all ages. Activities of all but the neuraminyltransferase decrease with age. The relatively greater neuraminyltransferase activity is consistent with increased incorporation of sialic acid into secreted mucins over the same age span. Conversely, diminution of fucosyltransferase relative to galactosyltransferase activity may contribute to the lower fucose content and lower blood group A activity of mucins secreted by mature ferret tracheas.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids↗