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Biomedical subjects

J S Patton

Publications and source records attributed to J S Patton.

51 records · Page 3Linked to original sources
PubMed1981

Inhibition of human pancreatic lipase-colipase activity by mixed bile salt-phospholipid micelles.

Mixed dihydroxy bile salt-phosphatidylcholine (PC) micelles can inhibit the hydrolysis of gum arabic-stabilized long-chain triglyceride emulsions by 10(-8) to 10(-9) M concentrations of human pancreatic lipase and colipase. Trypsin treatment of this colipase preparation did not reverse the inhibition, suggesting that procolipase, as a possible contaminant, was not the inhibitory factor. Human biliary phospholipid-cholesterol liposomes, isolated by gel filtration and redissolved in bile salt solutions, inhibited lipolysis to the same degree as solutions of bile salt containing purified PC. The degree of inhibition depended principally on the species of bile salt present (e.g., taurochenodeoxycholate greater than taurodeoxycholate greater than tauroursodeoxycholate greater than taurocholate). In the absence of bile salt, PC (0.4 mM) liposomes alone were not inhibitory over the physiological time range studied. Bile salt solutions of phosphatidylethanolamine or sphingomyelin also inhibited lipase activity, whereas those containing oleyl alcohol, oleyl aldehyde, oleic acid, and lyso-PC did not. PC molecules were found to partition between the triglyceride emulsion interface and the bulk aqueous phase. Full reversal of inhibition occurred in the presence of phospholipase A2, which hydrolyzed the phospholipids to lysolecithin and fatty acids. Mixed bile salt-phospholipid micelles caused marked decrease in the binding of lipase and colipase to the triglyceride substrate and displaced the proteins into the aqueous phase. The results taken together suggest that colipase binds to certain bile salt-PC associations independent of whether the aggregates are located at the surface of a triglyceride particle as a monolayer or in the bulk aqueous phase as mixed micelles.

Bile Acids and Salts↗
PubMed1979

Watching fat digestion.

During fat digestion a number of physicochemical events can be seen directly by light microscopy. Under simulated physiological conditions, hydrolysis of emulsified fat droplets by human pancreatic lipase in the presence of colipase and bile salt micelles proceeds with the sequential formation of two visible product phases. A lamellar liquid crystalline or crystalline phase containing calcium and ionized fatty acid forms first; this is followed by the production of a "viscous isotropic" phase composed predominantly of monoglycerides and protonated fatty acids.

Bile Acids and Salts↗
PubMed1978

Lipase-colipase interactions during gel filtration. High and low affinity binding situations.

The interaction of porcine pancreatic lipase and colipase was studied during gel filtration in columns eluted with a variety of buffers. High and low affinity binding situations were observed under different conditions. Low affinity binding could only be detected at the high lipase-colipase concentrations encountered during batch purification (10(-3)-10(-4) M). Even in this situation the rapid dissociation of the weak complex during filtration resulted in considerable separation of the two proteins. High affinity binding of lipase to colipase was observed at protein eluant concentrations as low as 10(-8) M on columns equilibrated with oleic acid-taurodeoxycholate mixed micelles. This binding did not take place on columns equilibrated with simple bile salt and mixed phosphatidylcholine-cholesterol-bile salt micelles. Colipase alone exhibited strong binding to phosphatidylcholine and fatty acid mixed bile salt micelles when applied together in a sample on columns eluted with pure bile salt micelles, lipase did not. The relevance of the high affinity complex to the lipase . colipase . substrate complex is discussed.

Animals↗
PubMed1977

Partial characterization of the bile salt-dependent triacylglycerol lipase from the leopard shark pancreas.

Leopard shark triacylglycerol lipase has been characterized as a crude pancreatic preparation. The enzyme demonstrated an absolute requirement for trihydroxy bile salts for activity with natural bile salts of the shark giving a 4-fold greater stimulation of activity than pure sodium taurocholate. Bile salts also protected the enzyme from apparent inactivation by p-chloromercuribenzoate and trypsin treatment. The shark lipase demonstrated a temperature optimum of 36 degrees C and was rapidly inactivated at 50 degrees C even in the presence of bile salts. Divalent metal ions were required for activity with Ca2+ providing the greatest stimulation. At 22 degrees C, pH 8.5 and in the presence of natural bile salts, the apparent V was about 0.6 mumol fatty acid released/min per mg protein. The shark enzyme hydrolyzed over 90% of the fatty acids from trioleovylglycerol and methyl esters of pancreatic lipase-resistant fatty acids were hydrolyzed at the same rate as typical fatty acid methyl esters. Hydrolysis of triacylglycerol proceeded about ten-times faster than wax ester hydrolysis. The kinetic properties of the leopard shark enzyme were compared to other bile salt-dependent lipolytic enzymes. Pancreatic lipase activity was not detected.

Animals↗
PubMed1975

High levels of pancreatic nonspecific lipase in rattlesnake and leopard shark.

Hydrolysis of synthetic triglycerides by rattlesnake and leopard shark pancreatic enzymes revealed striking differences in specificity, depending on the presence or absence of sodium taurocholate. Without added sodium taurocholate the classical specificity of pancreatic lipase was expressed. Rattlesnake enzymes, in the presence of sodium taurocholate, attacked the unsaturated oleic acid in the 2-position of racemic glycerol-1-palmitate-2-oleate-3-stearate nearly twice as fast as either outside saturated fatty acid. In this instance, over 90% of the monoglyceride which accumulated were 1-monoglyceride. These results are attributed to very high levels of bile salt activated nonspecific lipase. Eight vertebrate species were compared. With the exception of the rattlesnake and leopard shark, the other species (3 elasmobranchs and 3 mammals) all exhibited low levels of nonspecific lipase, e.g. less than 5% hydrolysis of the 2-position of racemic glycerol-1-palmitate-2-oleate-3-stearate in the presence of sodium taurocholate.

Animals↗
PubMed1975

Specificity of digestive lipases in hydrolysis of wax esters and triglycerides studied in anchovy and other selected fish.

The physiological specificity of fat digestion in several species of marine fish was studied by incubating a variety of synthetic and natural lipid substrates in fish intestinal fluid. Wax ester and triglyceride hydrolyses were studied in vivo and in vitro. In vivo feeding studies showed triglyceride hydrolysis and reesterification in the gut occurred 4 times faster than wax ester metabolism. In vitro comparisons of wax and triglyceride lipolysis always showed triglycerides to be hydrolyzed faster than wax esters; however, wide variation in the ratio occurred among different batches of intestinal juice. Ca. 50% of the 2 monoglycerides formed in the lipolytic sequence were hydrolyzed. Esters of lipase resistent fatty acids (20:4 and 20:5) were cleaved faster than normal fatty acid esters (18:2 and 18:3). Two of the species studied, the northern anchovy, Engraulis mordax and the jack mackerel, Trachurus symmetricus, empty lipase(s) into their gall bladders and produce-phospholipid free bile.

Animals↗
PubMedDate not supplied

The pattern of lung injury induced after pulmonary exposure to tumor necrosis factor-alpha depends on the route of administration.

TNF-alpha is a protein elaborated by monocytes and macrophages in response to endotoxin. The in vivo consequences of TNF-alpha elaboration have been examined extensively after intravenous administration of TNF-alpha. Substantially less is known about the effects of TNF-alpha that may be generated locally by resident tissue phagocytes. We investigated the direct effects of TNF-alpha on lung tissue by administering large amounts of human TNF-alpha directly to the lung, either as an aerosol or as an intratracheal bolus. Rats were exposed to an aerosol containing several concentrations of TNF-alpha, resulting in retention of significant quantities of TNF-alpha. The histologic response to inhaled TNF-alpha was characterized by adherence of leukocytes to venular endothelium, endothelial cell disruption, and bronchovascular edema. After aerosol administration, however, there was no evidence of alveolar inflammation or edema. In contrast, intravenous administration of large amounts of human TNF-alpha, at a dose that produced a lung content of TNF-alpha similar to that produced after high-concentration aerosol exposure, resulted in severe alveolar injury and edema. Intravenous administration of TNF-alpha did not result in the bronchovascular changes seen after inhalation. To ensure that sufficient quantities of TNF-alpha were being delivered to the lung, TNF-alpha was given as an intratracheal bolus to rats. This led to measurable absorption, but the spectrum and severity of lung injury was similar to the group that received TNF-alpha as an aerosol. We conclude that in rats, the pulmonary response to the injurious effects of TNF-alpha differ, depending on whether the TNF-alpha is delivered to the air or blood side of the alveolar capillary barrier.

Absorption↗
PubMedDate not supplied

Absorption of human growth hormone from the rat lung.

Recombinant methionyl human growth hormone (hGH) was administered intratracheally to adult rats, and serum concentrations of immunoreactive hGH were measured for up to 24 h. The mean absolute bioavailability was approximately 36% after 18h and was similar for doses of 0.75, 1.5, and 3 mg/kg. Peak serum hGH concentrations occurred at approximately 6 h after dosing. Tritiated hGH (3H-hGH) was used to follow the clearance of hormone from the lungs. Disappearance was linear with time and by 24 h approximately 70% of the radioactivity was gone from the lungs (elimination half-life = approximately 10.5h). Monomeric and aggregated hGH appeared to account for the majority of the residual 30% of radioactivity. Immunohistochemical localization of hGH in the alveoli suggested that the hormone was concentrated in a thin layer at the air-epithelial boundary. Pulmonary macrophages, which also stained for hGH, probably degrade hGH and thus account for some loss of material in the lungs. These studies suggest that the lung may be an alternative route for systemic delivery of recombinant proteins which are currently delivered by injection.

Absorption↗
PubMedDate not supplied

Induction of monooxygenase activity in the intestine of spot (Leiostomus xanthurus), a marine teleost, by dietary polycyclic aromatic hydrocarbons.

The response of intestinal monooxygenases to dietary polycyclic aromatic hydrocarbon (PAH) exposure was evaluated in spot (Leiostomus xanthurus), a marine teleost fish. Ethoxyresorufin O-deethylase (EROD) and aryl hydrocarbon hydroxylase (AHH) activities were highest in the pyloric caeca and in the proximal half of the intestine. Intestinal microsomes from fish given control diets had very low levels of EROD and AHH activities relative to those in liver. After exposure to a diet containing 10 mg of 3-methylcholanthrene/kg of food, the levels of intestinal EROD and AHH activities increased 36-fold and 17-fold, respectively, such that intestinal monooxygenase activity exceeded that of the liver, which was not induced by this treatment. A significant increase in intestinal monooxygenase activity occurred in fish receiving dietary benzo[a]pyrene (BP) at concentrations as low as 10 micrograms of BP/kg food. A 5-fold increase in intestinal AHH and EROD activities was observed within 3 hr after administration of dietary BP. A plateau in gut monooxygenase activity occurred after approximately 3 days of PAH exposure; these activities decreased to control levels within 3 days after replacing the PAH diet with the control diet. Starvation resulted in disappearance of detectable monooxygenase activity. Monoclonal antibody (MAB 1-12-3) against the major PAH-inducible cytochrome P-450 (P-450E) in the liver of the marine teleost (Stenotomus chrysops) [Park et al. Arch. Biochem. Biophys. 249, 399 (1986)] recognized a single protein band in intestinal microsomes, with Mr near 54,000, which we conclude is the spot counterpart to cytochrome P-450E.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗