The effect of bile, bile acids and detergents on calcium absorption in the chick

1. Bile from rachitic or normal chicks causes an immediate increase in the intestinal absorption of soluble calcium in rachitic and vitamin D(3)-treated chicks as tested in vivo by intestinal-loop and oral-dosing methods. 2. This effect is apparently solely due to the taurine-conjugated bile acids present in the bile and is independent of the action of vitamin D. 3. Chick bile and bile acids can increase the solubility and the absorption of calcium presented as sparingly soluble calcium hydrogen phosphate. 4. In addition, bile is necessary to some extent at least for the intestinal absorption of vitamin D(3) in the chick and this would indirectly enhance the absorption of calcium. 5. Thus bile is capable of a threefold action in the absorption of calcium in the chick. It is suggested that the direct action on sparingly soluble forms of calcium is of considerable physiological importance since most of the calcium in the normal bird's diet would be in this form. 6. Bile acids enhance the absorption of calcium in all regions of the small intestine of the chick. 7. Of a range of bile acids and detergents tested for enhancement of calcium absorption, various taurine-conjugated bile acids and sodium lauryl sulphate, an anionic detergent, are effective. A non-ionic detergent (Tween 80) and a cationic detergent (Zephiran) are without effect. 8. The ability of a substance to increase directly the intestinal absorption of soluble calcium appears to depend to some extent on an anionic detergent action, i.e. the ability to form a salt or complex soluble to some extent in both aqueous and lipid phases. 9. In chicks the immediate deposition of calcium ((45)Ca) in the bones closely reflects any increase in plasma calcium radioactivity regardless of the cause of the increase and regardless of the vitamin D(3) status. Although sodium lauryl sulphate can increase markedly the calcium absorption from the gut and the immediate deposition in the bones it has no significant effect on rickets. 10. Some of the implications of these findings are discussed.     

Bile and the absorption of strontium and iron

1. Strontium absorption was studied in vivo with loops of ileum in rachitic chicks and found to be increased by vitamin D(3), bile salts and sodium lauryl sulphate. 2. Bile salts and sodium lauryl sulphate rendered strontium soluble in butanol-benzene (1:1, v/v). 3. Bile was not concerned in the absorption of iron in rats from its water-soluble form, ferrous sulphate. 4. Ligation of the bile ducts in rats caused a decrease in the absorption of iron presented as its sparingly soluble phosphate. 5. The effect of bile on cation absorption is discussed.     

Bile acid solubility and precipitation in vitro and in vivo: the role of conjugation, pH, and Ca2+ ions.

The principles governing the in vitro solubility of the common natural conjugated and unconjugated bile acids and salts in relation to pH, micelle formation, and Ca2+ concentration are considered from a theoretical standpoint and then correlated first with experimental observations on model systems and second with the formation of precipitates containing bile acids in health and disease. In vitro, taurine-conjugated bile acids are soluble at strongly acidic pH; glycine-conjugated bile acids are poorly soluble at moderately acidic pH; and many of the common, natural unconjugated bile acids are insoluble at neutral pH. For both glycine-conjugated and unconjugated bile acids, solubility rises exponentially, with increasing pH, until the concentration of the anion reaches the critical micellization concentration (CMC) when micelle formation occurs and solubility becomes practically unlimited. In vivo, in health, conjugated bile acids are present in micellar form in the biliary and intestinal tract. Unconjugated bile acids formed in the large intestine remain at low monomeric concentrations because of the acidic pH of the proximal colon, binding to bacteria, and absorption across the intestinal mucosa. In diseases in which proximal small intestinal content is abnormally acidic, precipitation of glycine-conjugated bile acids (in protonated form) occurs. Increased bacterial formation of unconjugated bile acids occurs with stasis in the biliary tract and small intestine; in the intestine, unconjugated bile acids precipitate in the protonated form. If the precipitates aggregate, an enterolith may be formed. In vitro, the calcium salts of taurine conjugates are highly water soluble, whereas the calcium salts of glycine conjugates and unconjugated bile acids possess limited aqueous solubility that is strongly influenced by bile acid structure. Precipitation occurs extremely slowly from supersaturated solutions of glycine-conjugated bile acids because of metastability, whereas super-saturated solutions of unconjugated bile acids rapidly form precipitates of the calcium salt. In systems containing Ca2+ ions and unconjugated bile acids, pH is important, since it is the key determinant of the anion concentration. For bile acids with relatively soluble calcium salts (or with a low CMC), the concentration of the anion will reach the CMC and micelles will form, thus precluding formation of the insoluble calcium salt. For bile acids, with relatively insoluble calcium salts (or with a high CMC), the effect of increasing pH is to cause the anion to reach the solubility product of the calcium salt before reaching the CMC so that precipitation of the calcium salt occurs instead of micelle formation.(ABSTRACT TRUNCATED AT 400 WORDS)     

Absorbability of calcium from calcium-bound phosphoryl oligosaccharides in comparison with that from various calcium compounds in the rat ligated jejunum loop

Calcium-bound phosphoryl oligosaccharides (POs-Ca) were prepared from potato starch. Their solubility and in situ absorbability as a calcium source were investigated by comparing with the soluble calcium compounds, calcium chloride and calcium lactate, or insoluble calcium compounds, calcium carbonate and dibasic calcium phosphate. The solubility of POs-Ca was as high as that of calcium chloride and about 3-fold higher than that of calcium lactate. An in situ experiment showed that the intestinal calcium absorption rate of POs-Ca was almost comparable with that of the soluble calcium compounds, and was significantly higher (p<0.05) than that of the insoluble calcium groups. Moreover, the total absorption rate of a 1:1 mixture of the calcium from POs-Ca and a whey mineral complex (WMC) was significantly higher (p<0.05) than that of WMC alone. These results suggest that POs-Ca would be a useful soluble calcium source with relatively high absorption in the intestinal tract.     

Computational Oral Absorption Simulation for Low‐Solubility Compounds

Bile micelles play an important role in oral absorption of low‐solubility compounds. Bile micelles can affect solubility, dissolution rate, and permeability. For the pH–solubility profile in bile micelles, the Henderson–Hasselbalch equation should be modified to take bile‐micelle partition into account. For the dissolution rate, in the Nernst–Brunner equation, the effective diffusion coefficient in bile‐micelle media should be used instead of the monomer diffusion coefficient. The diffusion coefficient of bile micelles is 8‐ to 18‐fold smaller than that of monomer molecules. For permeability, the effective diffusion coefficient in the unstirred water layer adjacent to the epithelial membrane, and the free fraction at the epithelial membrane surface should be taken into account. The importance of these aspects is demonstrated here using several in vivo and clinical oral‐absorption data of low‐solubility model compounds. Using the theoretical equations, the food effect on oral absorption is further discussed.     

Effects of bile salts on the adsorption of a hydrophobic mutagen to dietary fiber

The effect of the bile salts, sodium cholate, deoxycholate, glycocholate and taurocholate, on the solubility in aqueous solution of the hydrophobic, environmental mutagen, 1,8-dinitropyrene (DNP), was examined. In the absence of bile salts, the DNP appeared to precipitate out of solution, whereas bile salts at a concentration of greater than or equal to 4 mM maintained the DNP in solution. In the presence of the model dietary fiber, alpha-cellulose, the DNP absorbed to this preferentially. Bile salts reduced this adsorption at low alpha-cellulose levels, but had little effect at high alpha-cellulose levels. The implication of these results is that bile salts have solubilising properties that could affect the distribution of hydrophobic molecules, including mutagens, in the digestive tract.     

THE EFFECT OF THE H ION CONCENTRATION ON THE AVAILABILITY OF IRON FOR CHLORELLA SP

The data obtained in these experiments indicate clearly that unless the necessary precautions are taken to keep the iron of the culture medium in solution the results obtained by varying the H ion concentration will not represent the true effect of this factor on growth. The availability of iron in nutrient solutions has been the subject of numerous recent investigations and it is now known that iron is precipitated at the lower hydrogen ion concentrations, that the iron of certain iron salts is less likely to be precipitated than that of others, and that certain salts of organic acids tend to keep the iron in solution. In general, ferric citrate seems to be the most favorable source of iron. In addition to chemical precipitation, however, it is also possible for the iron to be removed by adsorption on an amorphous precipitate such as calcium phosphate. As this precipitate is frequently formed when nutrient solutions are made alkaline, this may account for the discordant results reported in the literature as to the availability of certain forms of iron. By omitting calcium from the culture solution iron can be maintained in a form available for growth in alkaline solutions by the addition of sodium citrate. In such solutions the maximum growth of Chlorella occurred at pH 7.5. The alkaline limit for growth has not been established as yet. In investigating the availability of iron at varying concentrations of the hydrogen ion, changes in the pH value of the solution during the course of an experiment should also be taken into account. This is especially important in unbuffered solutions. The differential absorption of the ions of ammonium salts may cause a marked increase in the hydrogen ion concentration, which in turn will cause an increase in the solubility of iron. In strongly buffered solutions as used in these experiments this effect is slight.     

Relationships between bile salts hydrophilicity and phospholipid composition in bile of various animal species

Bile salts and phospholipids from bile of chicken, dog, sheep, rat, ox, pig, guinea-pig and man were analyzed by high-performance liquid chromatography. Bile salts showed marked differences in their hydrophilic properties, owing to hydroxyl structure and type of conjugation. Phospholipids were generally similar, containing 90-95% of phosphatidylcholine which was made of molecular species containing palmitic acid in the sn-1 position. The comparative analysis of bile salts and phosphatidylcholines profile demonstrated that bile salts hydrophilicity influences the quantity of phosphatidylcholine in bile but not the quality.     

Solution properties of sulfated monohydroxy bile salts. Relative insolubility of the disodium salt of glycolithocholate sulfate

Physical-chemical properties of the major sulfated monohydroxy bile salts of man are described. In general, the sulfates are significantly more water-soluble than the non-sulfated species as a result of lower critical micellar temperatures, high aqueous monomeric solubilities and critical micellar concentrations. Nevertheless, at 37 degrees C the disodium salt of glycolithocholate sulfate, the major monohydroxy bile salt of man is not more soluble than its non-sulfated form. Since aqueous solubility correlates inversely with the cholestatic potential of bile salts, our results suggest that this sulfate may be potentially hepatoxic. Micellar solubility of phosphatidylcholine and cholesterol by the majority of non-sulfated and sulfated monohydroxy bile salts is slight. Nonetheless, phosphatidylcholine is very well solubilized by taurolithocholate sulfate but cholesterol solubility is not increased appreciably. Cholesterol saturation in model bile systems of taurochenodeoxycholate and phosphatidylcholine is impaired by the addition of sulfated lithocholate conjugates but with physiological bile salt compositions this reduction is not significant.     

The effect of phosphate withdrawal by plant and by an anion-exchange resin on the phosphate potential of a soil

Summary Varying degrees of phosphate depletion by plants and AER were brought about in two acid soils. After depletion, the monocalcium phosphate potential was determined in soil suspended in dilute CaCl₂ solution.The results showed that phosphate depletion either by plants or AER did not change the phosphate potential of a soil. It seemed that some of the labile phosphate removed by plants was replaced by a mobilization of non-labile soil phosphate fraction in the soil.The constancy of the phosphate potential indicated that the concentration of phosphate in the soil solution was controlled by the solubility of sparingly soluble soil phosphates. The data implied that it was the solubility of hydroxyapatite which determined the phosphate potential of both soils.     

Coprecipitation modulates the solubility of minerals in bovine milk

The solubilities of zinc, iron, copper, magnesium, calcium, inorganic phosphate, and citrate in milk decreased when acidic milk preparations were neutralized. In decaseinated bovine milk soluble zinc, iron, and copper were reduced 90%, 60%, and 50%, respectively, as the pH was raised from 4 to 7. Simultaneous precipitation of minerals and citrate was confirmed by analysis of washed precipitate. We propose that the diminished solubilities of zinc, iron, copper, magnesium, and citrate are linked to the precipitation of calcium phosphate through one or more mechanisms of coprecipitation. Such control on mineral solubility may have an impact upon mineral absorption from milk.     

Effect of pH on the speciation and solubility of divalent metals in human and bovine milks

Computer models estimated the ligand speciation and solubility of calcium, magnesium, zinc, and copper over a pH range for low molecular weight fractions characteristic of either human or bovine milks. Above pH 4 calcium is the only metal predicted to precipitate. Most of the remaining soluble calcium, magnesium, and zinc should be complexed with citrate. The solubility of calcium, magnesium, and zinc in human and bovine milks was measured experimentally from pH 2 to 7. The solubility of all three metals decreased as the pH increased. Calcium and zinc were soluble over a narrower pH range in bovine milk than in human milk. Increasing the levels of either calcium or inorganic phosphate alone in decaseinated human milk did not affect the solubility of zinc, but when both calcium and inorganic phosphate were added at levels comparable to bovine milk the solubility of zinc decreased at the higher pH's. The decreased solubility of zinc in skimmed milks in pH's characteristic of the small intestine is likely due to coprecipitation of zinc with calcium phosphate--a reaction not predicted for milk systems from known chemical solubility product data.     

Gall Stones in Patients with Disorders of the Terminal Ileum and Disturbed Bile Salt Metabolism

In a combined historical and radiological survey gall stones were found in 23 out of 72 patients with disorders of the terminal ileum (31·9%). This is four to five times the expected incidence of cholelithiasis. In these patients the incidence of gall stones was not related to age, but did, however, increase with increased duration of the ileal disorder. Glycine/taurine conjugation ratios of bile salts in the bile were abnormally high in 10 out of 11 patients with ileal disorders. Both impaired bile salt recirculation and absorption of poorly soluble bacterially degraded bile salts are possible causes of cholelithiasis in these patients.     

Inhibition of Injured Escherichia coli by Several Selective Agents

A population of Escherichia coli ML30 cells was exposed to a quaternary ammonium compound, and injury to the cells was measured by a comparison of counts on Trypticase Soy Agar and Violet Red Bile Agar. Substantial injury could not be detected with a minimal medium. The ingredients of Violet Red Bile Agar were tested against damaged cells. The bile salts mixture alone in the medium prevented as many injured cells from growing as did any combination of the selective agents and inhibited as many injured bacteria as were inhibited by Violet Red Bile Agar itself. These dyes and salts were similarly assayed in minimal agar, and comparable results were obtained. Individual bile salts and other potential selective agents were added to the minimal medium, and the media were tested for inhibition of injured E. coli. Sodium deoxycholate was the bile salt most inhibitory to damaged E. coli cells.     

Effect of distal enterectomy on cholesterol and bile salt levels in the rat

The effect of 50% or 80% distal enteroctomy on cholesterol and bile salt levels in male Wistar rats have been investigated. Short time measurements showed that serum cholesterol levels were maximal after 20 days from 50% intestinal resection and after 10 days from 80% intestinal resection. This increase was maintained in 50% resected rats 1 and 5 months after operation, whilts in 80% resected group the values became normal. Portal blood and bile cholesterol levels remain almost normal except 5 months after 50% intestinal resection. Bile salt concentration and bile salt output in the bile decrease after 1 and 5 months from 50% intestinal resection and after 1 month from 80% intestinal resection. These results together with data of fecal loss of bile salts indicate that in 50% resected rats new steady states have been reached, with low levels of bile salts in the bile. One month after 80% resection the fecal loss of bile salts was so high that the conversion of cholesterol into bile salts was increased. After 5 months from 80% resection values in serum and bile were almost normal suggesting either an increase in extrahepatic cholesterol synthesis or a partial prevention of fecal loss that can be explained by the observed caecal enlargement.     

Structural basis for bile salt inhibition of pancreatic phospholipase A2

Bile salt interactions with phospholipid monolayers of fat emulsions are known to regulate the actions of gastrointestinal lipolytic enzymes in order to control the uptake of dietary fat. Specifically, on the lipid/aqueous interface of fat emulsions, the anionic portions of amphipathic bile salts have been thought to interact with and activate the enzyme group-IB phospholipase A2 (PLA2) derived from the pancreas. To explore this regulatory process, we have determined the crystal structures of the complexes of pancreatic PLA2 with the naturally occurring bile salts: cholate, glycocholate, taurocholate, glycochenodeoxycholate, and taurochenodeoxycholate. The five PLA2-bile salt complexes each result in a partly occluded active site, and the resulting ligand binding displays specific hydrogen bonding interactions and extensive hydrophobic packing. The amphipathic bile salts are bound to PLA2 with their polar hydroxyl and sulfate/carboxy groups oriented away from the enzyme's hydrophobic core. The impaired catalytic and interface binding functions implied by these structures provide a basis for the previous numerous observations of a biphasic dependence of the rate of PLA2 catalyzed hydrolysis of zwitterionic glycerophospholipids in the presence of bile salts. The rising or activation phase is consistent with enhanced binding and activation of the bound PLA2 by the bile salt induced anionic charge in a zwitterionic interface. The falling or inhibitory phase can be explained by the formation of a catalytically inert stoichiometric complex between PLA2 and any bile salts in which it forms a stable complex. The model provides new insight into the regulatory role that specific PLA2-bile salt interactions are likely to play in fat metabolism.     

The Mechanism of Enterohepatic Circulation in the Formation of Gallstone Disease

Bile acids entering into enterohepatic circulating are primary acids synthesized from cholesterol in hepatocyte. They are secreted actively across canalicular membrane and carried in bile to gallbladder, where they are concentrated during digestion. About 95 % BAs are actively taken up from the lumen of terminal ileum efficiently, leaving only approximately 5 % (or approximately 0.5 g/d) in colon, and a fraction of bile acids are passively reabsorbed after a series of modifications in the human large intestine including deconjugation and oxidation of hydroxy groups. Bile salts hydrolysis and hydroxy group dehydrogenation reactions are performed by a broad spectrum of intestinal anaerobic bacteria. Next, hepatocyte reabsorbs bile acids from sinusoidal blood, which are carried to liver through portal vein via a series of transporters. Bile acids (BAs) transporters are critical for maintenance of the enterohepatic BAs circulation, where BAs exert their multiple physiological functions including stimulation of bile flow, intestinal absorption of lipophilic nutrients, solubilization, and excretion of cholesterol. Tight regulation of BA transporters via nuclear receptors (NRs) is necessary to maintain proper BA homeostasis. In conclusion, disturbances of enterohepatic circulation may account for pathogenesis of gallstones diseases, including BAs transporters and their regulatory NRs and the metabolism of intestinal bacterias, etc.     

Phospholipid lamellae are cholesterol carriers in human bile

Cholesterol solubility and precipitation in bile are major factors in the pathogenesis of cholesterol gallstones. At present, mixed micelles and phospholipid vesicles are considered to be the only cholesterol carriers in bile. In this study we present evidence showing that phospholipid lamellae are major cholesterol carriers in human bile. Lamellae are a known aggregational form in pure phospholipid model systems. In the present study, lamellae were demonstrated by electron microscopy after negative staining and by small-angle X-ray diffraction in all human gallbladder bile samples examined. During diffraction experiments, cholesterol was found to crystallize from these lamellae. Cholesterol carriers in bile were separated by high-resolution chromatography and by prolonged ultracentrifugation. Lamellae were shown to solubilize most of the biliary cholesterol; vesicles solubilized a lesser amount; while micelles solubilized only a minor portion. Our data suggest that phospholipid aggregates are the main cholesterol carriers in bile. Bile salts may control the equilibrium between the various aggregational forms of cholesterol-carrying phospholipids.     

Bile salts in submicellar concentrations promote bidirectional cholesterol transfer (exchange) as a function of their hydrophobicity

Cholesterol, despite its poor solubility in aqueous solutions, exchanges efficiently between membranes. Movement of cholesterol between different subcellular membranes in the hepatocyte is necessary for assembly of lipoproteins, biliary cholesterol secretion, and bile acid synthesis. Factors which initiate and facilitate transfer of cholesterol between different membranes in the hepatocyte are incompletely understood. It is known that cholesterol secretion into the bile is linked to bile salt secretion. In the present study, we investigated the effects of bile salts of different physicochemical properties at submicellar concentrations (150- 600 microM) on the transfer of [14C]cholesterol from hepatocytes, or crude hepatocellular membranes (donors), to rat high density lipoproteins (acceptor). Bile salts included taurine conjugates of ursodeoxycholic acid (TUDCA), hyodeoxycholic acid (THDCA), cholic acid (TCA), chenodeoxycholic acid (TCDCA), and deoxycholic acid (TDCA). High density lipoprotein (HDL) was separated from hepatocellular membranes and the transfer of [14C]cholesterol from the membranes to HDL was quantitatively determined. In the absence of HDL, [14C]cholesterol remained confined to the membrane fraction. Following addition of HDL, [4-14C]cholesterol in the HDL fraction increased linearly over time. Addition of hydrophilic bile salts (TUDCA and THDCA) increased transfer of [4-14C]cholesterol to HDL only minimally. By contrast, more hydrophobic bile salts stimulated transfer of labeled cholesterol to HDL, and their potency increased in order of increasing hydrophobicity (TCA less than TCDCA less than TDCA). Both for single bile salts and mixtures of bile salts at a total bile salt concentration of 0.30 mM, the rate of cholesterol transfer exhibited a strong linear correlation with a bile salt monomeric hydrophobicity index (r = 0.95; P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)